Cover photo legend, in Bill’s own words:

This remains one of my favourite shots, although it is an old slide and this is the best digital scan | have of it (but of

woefully low res). It was taken about 25+ years ago, and | was driving to work when | saw these flowers in bloom beside

Port Elizabeth airport. | had an image of a cobra rearing in front of them, the Port Elizabeth Snake Park had just got

a beautiful Cape Cobra in from the Northern Cape, and so | asked Rob Hall to come and help manipulate the snake.

| didn't have a long lens and so had to lie on my belly with a 55mm Nikkon with 1.4 convertor. | used in-fill flash, held

by Rob about 1m away and to soften the deep shadow under the snake's belly. | kept shuffling forward to get a more

dramatic pose and had taken several shots when the snake disappeared from the viewfinder. Rob was standing to the

side holding the flash and also a snake stick to ward off the cobra. When the snake disappeared | instinctively rolled

back, heard Rob shout "Shit, that was fast!", and the snake bit the camera body about 6cm from my shutter finger. A

bead of venom glistened on the camera body. Looking through the lens | had lost all sense of distance and simply got

too close to the snake. It remains the closest I've come to a snakebite. Technically the picture works because the snake

is alert but its mouth is shut and it is not looking straight at the camera. It therefore doesn't appear too threatening,

allowing viewers to admire what remains my favourite snake. Bill Branch

Official journal website:

amphibian-reptile-conservation.org

Amphibian & Reptile Conservation

13(2) [Special Section]: i-—xxix (e186).

Compilation of personal tributes to William Roy Branch

(1946-2018): a loving husband and father, a good friend,

and a mentor

1*Werner Conradie, *7Michael L. Grieneisen, and *Craig L. Hassapakis (Editors)

'Port Elizabeth Museum (Bayworld), P.O. Box 13147, Humewood 6013, SOUTH AFRICA *School of Natural Resource Management, George

Campus, Nelson Mandela University, George 6530, SOUTH AFRICA *Department of Land, Air and Water Resources, University of California,

Davis, California 95616, USA *Amphibian & Reptile Conservation (amphibian-reptile-conservation.org) and Amphibian Conservation Research

Center and Laboratory (ACRCL), 12180 South 300 East, Draper, Utah 84020-1433, USA

Abstract.—Personal contributions to William “Bill” Roy Branch by famly members and colleagues: Colin

Tilbury, Alan Channing, Dot Hall (Pitman, Basson), Rick Shine, James B. Murphy, Luke Verburgt, Julian Bayliss,

Michael F. Bates, Pedro Vaz Pinto, Kirsty Kyle, Krystal Tolley, Mzi Mahola, Brian J. Huntley, Roger Bills, Johan

Marais, Mark-Oliver Rodel, Paul H. Skelton, Aaron M. Bauer, Stephen Spawls, Andrew Turner, Ernst H.W. Baard,

Amber Jackson, Margaretha Hofmeyr, Jens Reissig, Harold Braack, Atherton de Villiers, Marius Burger, Mike

Raath, Werner Conradie, and Martin J. Whiting.

Keywords. Influence, contributions, farewell, African herpetology, history, researcher

Citation: Conradie W, Grieneisen ML, Hassapakis CL (Editors). 2019. Compilation of personal tributes to William Roy Branch (1946-2018): a loving

husband and father, a good friend, and a mentor. Amphibian & Reptile Conservation 13(2) [Special Section]: i—xxix (e186).

tion 4.0 International (CC BY 4.0): https://creativecommons.org/licenses/by/4.0/], which permits unrestricted use, distribution, and reproduction in any

medium, provided the original author and source are credited. The official and authorized publication credit sources, which will be duly enforced, are

as follows: official journal title Amphibian & Reptile Conservation; official journal website: amphibian-reptile-conservation.org.

Received: 5 August 2019; Accepted: 5 August 2019; Published: 10 September 2019

On 14 October 2018, William Roy Branch, or simply

Bill as he was known to most, passed away after a short

struggle with motor neuron disease. He was not only

one of South Africa’s most well-known and respected

herpetologists, but also a dedicated husband, a father, a

good friend, and a mentor to so many of us. We have

taken this opportunity to collate personal tributes from

family, friends, and colleagues, to showcase the influence

Bill had on our lives and careers.

Tributes from family members

Donve Branch (Bill’s wife)

Bill was an amazing man with a huge passion for life.

When we married I introduced Bill to the world of

pots and potters, and he introduced me to the world of

reptiles and herpetologists. Very different worlds, but

they became one we both loved. Over the years I was

privileged to meet and host many of you. If I sometimes

looked stunned when you arrived at our door, please

forgive me. Bill very often failed to tell me we would

be having a guest. Together we started to collect art,

succulents, and books. None of which we could afford,

Correspondence. * werner@bayworld.co.za

Amphib. Reptile Conserv.

but we couldn’t resist.

Bill was a family man who loved and was so proud of

his three sons. When we married his generous heart took

on my children and grandchildren with the same warmth.

Science was his passion which he loved to share. Bill in

lecture mode could not be halted. His sense of humour

was legendry. A kind, gentle man but also a humble

man. He never boasted of his achievements. In his later

years, these qualities made him so popular with National

Geographic travellers.

A man of huge intellect with a broad knowledge of

all things. A kind, generous, and wonderful man. Truly a

real mensch. I was so proud to be his wife. He is greatly

missed.

James Vlok (Bill’s stepson)

Bill Branch was a man of passion for his craft and

natural science. He was an adventurer and an explorer;

a man who inspired motivation and discovery of the

world around us. He could keep you interested with a

keen knowledge and a sense of humour that would have

you laughing and learning. He will be sorely missed by

family and colleagues alike.

September 2019 | Volume 13 | Number 2 | e186

Tributes to William Roy Branch (1946-2018)

Christian Vlok (Donveé’s grandson)

Gumps told me so many interesting stories about his

trips. He gave me my first Masai machete and a lizard. I

knew I could ask him anything and he would know the

answer. I will miss my Grandpa Gumps so much.

Analeah Vlok (Donve’s granddaughter)

I loved Gumps because he taught me so many things.

He taught me which plants I can or can't eat, and about

snakes and frogs, which I love. I miss him and every time

I go into his room I think of him.

Jenny Vlok (stepdaughter-in-law)

Bill, to look at all things herpetological on a daily basis

and know that I can't ask you any more questions about

it, fills my heart with such sadness. You were so patient

in your explanations, always interesting and funny. With

your mismatched socks and wild hair, your fancy salads

and poor man's capers, hilarious Easter egg hunts with

a difference, cheeky Halloween surprises and Christmas

gifting, not only were you an Amazing scientist but also

an inspiration, and a motivator, allowing my children

to be knowledge bearers and researchers in their own

environment. We love you Dear Bill, and will miss you

always.

Nicole Kingston (Bill’s stepdaughter)

Bill was a rock, a voice of reason, and a safe place and

so loved. I am so truly privileged to have known him,

and am a better person for it. His kindness, empathy, and

wicked sense of humour will not be forgotten.

Oliver Kingston (Donve’s grandson)

Grandpa made me laugh lots and 1f you wanted to know

anything he was the person to ask.

Will Kingston (Donve’s grandson)

He was kind and knew a lot about snakes.

Tributes from friends and colleagues

Colin Tilbury

KwaZulu-Natal, South Africa

It was early in 1980. After a year as a junior medical officer

at Ngwelezana Hospital in KZN [KwaZulu-Natal], I had

collected a series of cases of snakebites from Atractaspis

bibronii and the Mozambique Spitting Cobra. With the

data in hand I had approached Alan Channing, the then-

chairman of the Herpetological Association of Africa,

for comments. Being more of the toad persuasion,

Alan suggested that I contact his colleague Bill Branch,

the incumbent curator of herpetology at the PE [Port

Elizabeth] Museum, who had shown more than just a

passing interest in snakes and snakebite, and might be in

Amphib. Reptile Conserv.

a better position to help me.

I wrote to Bill and offered to assist with any affairs of

the herpetological kind from Zululand. Bill wrote back

immediately, expressing a keen interest in the snakebite

data and also wondering if I might be able to collect some

of the local Pelusios for karyotyping. We met for the

first time a few months later. Bill was visiting Durban,

and Sarah and I arranged to meet him at the British

Middle East Indian Sporting and Diners’ Club near the

Greyville race course, to sample the local curries. A truly

memorable evening (I still have intermittent diarrhoea). I

think that I may also have introduced Bill to the pleasures

of a good red wine—or was it vice versa?

And so began a friendship which lasted nearly 40

years.

Driven with a boundless energy and an amazing

zest for life, sharp wit, wry humour, and capacity for

sharing, Bill attracted people to him. Whether by active

involvement or by association, he had a lasting impact

on all those who encountered him. Bill adored the simple

things in life, and lived his life simply. He loved the

camping and field trips that were an integral part of his

work and which provided him with so much satisfaction.

An avid angler since childhood, he had pulled many a

carp from the rivers and dams of the Eastern Cape. Bill’s

laboratory and office in the Port Elizabeth Museum was

always a wondrous place to visit. Beyond the entrance

door which was plastered with a selection of humorous

“Bill” references, a mixture of chaos and creativity,

preserved snakes and lizards in piles, the air reeking

with alcohol, and Bill smiling happily. Bill and Donveé’s

lovely home in Port Elizabeth was in many ways an

extension of his beloved office at the museum. Of the

many enduring mental images that capture Bill’s essence

for me, are none more so than those of Bill at work in

his man-cave at home. More like a ‘control room,’ his

desk surmounted with massive computer screens and

surrounded on all sides—floor to ceiling—with books,

paintings, and photographs (including his all-time

favourite of the yellow Cape Cobra that had nearly bitten

him). Shelves were packed with w.1.p. files and books

with titles covering an eclectic array of topics from

tadpoles to volcanoes, fossils, sunbirds, euphorbias,

mesembs, and every conceivable reptile and amphibian

genre.

At home, but outside his study, every nook and cranny

was adorned with paintings and Donve’s beautiful

pottery. Each windowsill in the house was crammed with

weirdly-shaped, rare, and spiky plants. Their garden was

an indigenous plant paradise with a few thorny exotics,

a haven for birds and local wildlife where the largest

Palystes rain spiders in the world were free to roam—

although strangely I only ever saw them on the walls of

the guest bedroom. Theirs was clearly a home they loved

to live in and was always open to the many guests who

might pop in and stay over.

September 2019 | Volume 13 | Number 2 | e186

Conradie et al.

One could not know Bill and be unimpressed with

his amazing intellect. Bill read—-no—he devoured books

by the ton. I have never met anyone who had such a

command and broad understanding of natural history. He

could have been anything from botanist, ornithologist,

entomologist, mammologist, geologist, physicist—you

name it. The reality in fact, is that he was all of these

things and many more; such was the breadth and depth

of his knowledge. His intimate understanding of the

intricacies of natural history, the environment, and the

interconnected webs of life, filling in the dots on life’s

canvas one by one—or in Bill’s case, by the dozens at

a time.

In spite of his huge talents, he kept his feet firmly on

the ground and freely shared his knowledge and wisdom

with anyone who asked for advice or input. He was an

inspirational force to anyone and everyone who had the

privilege to know or work with him; a truly benevolent

gentle giant and an incredibly productive scientist. The

herpetological community around him was so privileged

to have him as a guide and mentor. In the decade following

his retirement from the museum, he worked as a specialist

guide for over 50 National Geographic touring parties.

These afforded him opportunities to continue to pursue

herps in many iconic African locations.

As a friend, Bill was caring, insightful, non-

judgmental, and always with a wonderful sense of

humour just bubbling beneath the surface. As a storyteller

he had few peers: in his clipped British accent with the

hint of a lisp mumble and a wry smile, he would gleefully

extol the excruciating agony of the many unfortunates

who became the subjects of his tales. Of course, these

often involved his hapless colleagues on the many field

trips that he made. Quick, dry, wicked, invariably veiled

in intrigue, he would construct the twists and turns of

his story to extract every molecule of humour. His punch

lines always immaculate.

Over the years, I spent a great deal of time outside the

borders of South Africa, but Bill always found time to

write and give updates on his projects and movements.

After the birth of our first child in London in July

1989, Sarah and I sent out a short notice of his birth

to a few friends and relatives, making reference to ‘the

discovery of a new species of the TILBURY genus found

lurking in the St Helier 's Labour Ward at precisely 03h45

hours on 25 July 1989. It is wriggly, pink all over, devoid

of scales and tail, and makes characteristic feeding cries

every 4 hours. It weighed 3.63 kg on discovery, and has

the features characteristic of the male sex. It has been

named Douglas Matthew.”

I left London a week after the event and headed back

to my job in Saudi Arabia. Shortly after my return to

Khamis Mushayt, I received a letter from Bill:

“Dear Colin,

Congratulations on the arrival of Douglas Matthew.

You must be looking forward to Sarah and DMT

Amphib. Reptile Conserv.

arriving at the end of the month, although I suppose

that throwing all the Cerastes out to make way for the

cot must be a bind. He will slow your globe-trotting

down a bit, but it will only be about 12 years before

he is useful in the field! Robbie and Matthew do all

the hard work in the field now, so they do have their

advantages.

[feel | must take exception to the new name however.

Looking through the London telephone directory I

came across three other references to Tilbury Douglas

Matthew (usually from the poorer eastern suburbs

besides the Thames). All had priority, some dating from

the early 1930's. Your new name is thus pre-occupied,

and according to strict nomenclatural rules (Int. Rules

Zool. Nomenclature, rev. ed. London 1986; page 25,

paragraph 3), becomes a strict junior homonym and

is invalid. As well as afflicting the young lad with a

used name, it is also incorrectly formed according

to the rules governing construction of names. Being

the first scientist (of truly international standing) to

have spotted this error, I claim my right to propose a

replacement name. I have chosen:

Tilburyanus inhirsutus arabicum Branch 1989

You will note that the Generic name is now correctly

Latinized and the ending is more appropriate

(being his most obvious feature for the moment!).

The specific epithet also refers to the sub-adult

plumage, while the sub-specific name is a traditional,

uninspiring geographical allocation. Knowing that he

is now correctly named, you may re-apply for birth

certificates, passports and driving licences etc.”

For a man who played with snakes, Bill had a simple

philosophy. Respect them and you won’t get bitten,

and as far as I know, apart from a single dry bite from

a Thelotornis, he never did. I remember the day that I

brought a small shiny black snake all the way from

the DRC [Democratic Republic of the Congo] to Bill’s

home, and proudly handed him the blue cotton bag that

contained the snake which I had carefully nurtured for

the previous month or so. Bill gleefully but carefully

opened the bag and peered inside. Then to my horror, he

inserted his hand into the bag to retrieve the snake.

I said ‘Whoa! Hang on there a minute; I just want to

get out my notebook and camera to record the first bite

from this unknown species of Atractaspis.’ Bill pulled

out his hand, the snake dangling limply between his

fingers. Rigor mortis had already worn off.

“You've killed it” I said.

IN Oe ete nit excuses Do you think it is a Norwegian

Blue?” (A joke that can only be appreciated by followers

of Monty Python).

But it was not only herps that Bill would talk about.

As much as he was a scientist, he was also a profoundly

loved family man who would talk with pride as much

about his loved ones as he would about his work. Give

him half a chance and he would talk for hours about his

September 2019 | Volume 13 | Number 2 | e186

Tributes to William Roy Branch (1946-2018)

sons Robbie, Matthew, and Tom and his new family—

Nicole, Anthony, and James.

It was in 2012 that Bill first realised that he was

only human after all, when he contracted malaria in

Mozambique. That nearly finished him off. He eventually

bounced back to his old self, but he was unable to dodge

the bullet of MND [motor neuron disease]. When Bill

contacted me in February 2018 to say that he had been

diagnosed, it felt as if a tree had fallen on me. I had the

good fortune to be able to spend several days with Bill

over the last months of his life; and to be able to share

memories of the good times, laugh together, discuss the

iniquities of life, and to acknowledge the simple fact that

we are all just fulfilling our biological destiny—albeit in

different ways.

One cannot write about Bill without acknowledging

the major part in his life that Donvé played, as his partner

and soul mate, and in turn appreciate the huge hole that

has been left behind by his passing in Donve’s life. In

one conversation we had, we both agreed that it was one

of the greatest privileges of life to be able to love and be

loved back unconditionally. I don’t think that anyone can

overestimate the enormity of this gift. She made him so

happy and in the end, so sad that his Dove would have

to endure the last days of his life with him in the state he

was in.

Bill asked me to sign as witness to his living will to

not be placed on any mechanical machine that would

prolong his life. As his MND advanced, even in the late

stages, in spite of his body being totally paralysed, his

mind was as lively as ever; as he fought day by day to

extract, utilise, and enjoy to the last moment every second

that was left to him. He was immensely saddened and so

disappointed that he had run out of time to complete all

the many projects that he was part of or had initiated.

His illness had quite literally pulled the rug out from

beneath his feet. I know that Bill handed over many

of these to colleagues to finish—we should make him

proud. Even as he inexorably neared the end, he was so

brave in facing his fate. He could still make jokes about

this. He once compared himself as a likeness of the blue-

headed agamid that was named after him (Acanthocercus

Fig. 1. Cover image of Hyperolius raymondi used for Frogs and

other Amphibians of Africa (Photo: Bill Branch).

Amphib. Reptile Conserv.

branchi). Finally in the afternoon of 14 October, dulled by

the ever-increasing CO, levels, he finally and peacefully

breathed his last. The end to a magnificent life. His was

an act we could all learn something from.

More than anyone, Bill understood and appreciated the

fact that no one gets to live forever, but that everyone is

hopefully gifted with the opportunity to leave a footprint

embedded in the rocks of humanity—a footprint that will

endure with a permanent relevance to those who follow

one’s trail. Bill had big feet for such a small frame, and no

doubt we will be following his prints for many a year. I can

only say that I was privileged to know Bill, and even more

so, to think that he might have considered me to be a friend.

The memories of Bill will be enduring and he will

always be celebrated as one of the world’s leading

herpetologists of our time. He will be sorely missed and

long remembered.

Alan Channing

University of the Western Cape/North-West University,

South Africa

I met Bill at a herp meeting while he was working at

the Atomic Energy Board in the 1970s. He was hugely

enthusiastic and well-read. Later, I was happy to support

his application for the post of Herpetologist at the Port

Elizabeth Museum, when asked by the Director. We

undertook many field trips together, and for a while we

formed a collaboration for funding from the forerunner

of the National Research Foundation.

Although Bill and I worked on different groups, there

was always a lot of friendly banter between us. His sense

of humour was displayed on one field trip to northern

Namibia, when he offered to cook the potatoes, while I

prepared the meat. When it came time to eat, the potatoes

were still crunchy. Bill's response was to explain that that

was how they were cooked in Cornwall, and that it was a

classical culinary procedure!

I will miss Bill's insights and our regular email

exchanges. He provided a number of excellent photos

for the upcoming book Frogs and other Amphibians of

Africa, and was always willing to help, or offer a beer

and a meal, when I was in Port Elizabeth.

as - i =

Fig. 2. Bill p otographing a lizard in southern Angola, 18

January 2009 (Photo: Alan Channing).

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Conradie et al.

Dot Hall (Pitman, Basson)

Port Elizabeth Museum (Bayworld), South Africa

A flood of memories flow through my mind when I

reminisce on the very small part of Bill’s life I shared.

One in particular always makes me smile. When Bill

joined the Museum in 1978, he was a complete unknown.

We were observing his introduction to the staff with

interest: A quiet, rather serious little man? From his first

day he was a regular library user. He was passionate

about books. Each visit he made to it was a learning

experience for me. He freely shared his knowledge and

always stretched my way of thinking.

On one memorable morning, shortly after he had

joined the staff, all was quiet in the library when a strange

scuffing noise caught my attention. No-one was in the

library, so I put this observation down to my imagination

and continued working. The same noise recurred several

times till I eventually decided to investigate. There was

a solid counter that separated the librarian from those

using the library. I peered over this counter to find Bill

on all fours, crawling behind a large leguaan [varanid]

holding its tail and trying to direct it around the corner

to my desk. I guess it was being a little uncooperative

and his full attention was required for him to achieve his

goal—“Frighten this librarian out of her mind!!!” After

observing the scene for a short while I decided to launch

a surprise “attack” from the back and gave him a pinch

on his rear end. His reaction was marvellous. The leguaan

was let loose and his fright was complete.

Fig. 3. Bill Xerox-ing a puffadder to make counting of scales

easier, to the disgrace of the librarian (Photo: Dot Pitman).

Amphib. Reptile Conserv.

We both enjoyed sharing this amusing moment.

How many million more smiles has he given to the vast

number of people with whom he associated?

Rick Shine

Macquarie University, Australia

I first met Bill Branch on the morning of Tuesday the

5th of September 1989, at the British Museum of Natural

History (now the Natural History Museum London).

Like me, he had travelled to the UK to attend the First

World Congress of Herpetology, and like me, he took

advantage of the opportunity to visit the British Museum

of Natural History. Bill was looking for type specimens

of African herps, and I was attempting to track down

the reptile specimens that Charles Darwin collected in

Australia during the voyage of the Beagle. As we sat

and talked over lunch, I was astonished at Bill’s breadth

of knowledge about the African herpetofauna, and his

intimate familiarity with the scientific literature on those

animals. But I had no idea that we would end up as

collaborators on a major project.

Five years later, I took my first (and only!) sabbatical

from the University of Sydney. My wife Terri and I

had always wanted to see the famous game reserves of

southern Africa, and our oldest son was about to turn

12—after which time he would have to pay a full fare

on the airlines rather than half-price! So I contacted

Bill about the possibility of dissecting preserved snakes

in African museum collections for ecological data

leg

+, anche © al

AAT 4

£9

to look for reptiles, we never found any (Photo: Dot Pitman).

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Tributes to William Roy Branch (1946-2018)

POFADDER é

(gut contents, gonads, etc.) as I had done for snakes in

Australian museums over the previous years. Bill was

enthusiastic, but thought that the best collection might be

in Namibia, where over 1,500 snakes that had drowned

in an open canal (the Eastern Water Carrier) had been

preserved by the local wildlife authorities. And with the

first South African elections looming, and political unrest

likely as power shifted from ‘Afrikaaners’ to native

Africans, Namibia looked like a quieter, safer option than

South Africa for a family with 10-year-old and 2-year-

old children.

We flew to Namibia while Bill drove the museum’s

Kombi-van from Port Elizabeth to Windhoek to meet us.

And being Bill, he had a much better idea than staying in

Windhoek to dissect the snakes—instead, we piled them

into the van and took off for Gobabeb, where we could

enjoy the spectacular dunes in between long hours of

peering inside dead snakes. The Aussie team (me, Peter

Harlow, and Jonno Webb) peered inside the innards of

dead snakes and called out numbers, while Terri wrote

them all down into data-sheets. Bill carefully examined

every half-digested frog and reptile that came out of a

snake stomach, almost always managing to ID it, even if

he only had a few toes to work with. It was a happy and

effective team.

After we finished the Namibian snakes, my family

flew off to the USA while the rest of us drove down to

Pretoria to look at MORE snakes at the national museum

in Pretoria. It was a classic herp “road trip,” with frequent

detours to look for specific taxa (usually, so that Bill

could get a photograph for his field guide). We made an

obligatory stop at Poffadder (= “Puff Adder’) one of the

few towns named after a snake, near the border between

South Africa and Namibia. A photograph I took on the

town’s outskirts captures the relaxed joy of herpetological

zealots indulging their passions (Fig. 5). We worked

long hours in Pretoria, obtaining a mountain of data that

eventually translated into 15 papers on the natural history

of several major lineages of African snakes. We also

sampled the local beer and watched World Cup soccer

games at bars downtown—horrifying some of the locals

who were convinced that we would be mugged as we

walked the streets at night.

Throughout this first African adventure, Bill was

fantastic. Extraordinarily knowledgeable, with a vast

network of contacts, he made the project possible. We

talked long and often about everything from fishing

to the mysteries of bureaucracies and families—and

especially, about snakes. Hopping off a plane and looking

inside preserved specimens can generate a lot of data—

but it was Bill’s long experience that enabled us to put

that information into context. For many of the species

about which we wrote papers, I had never even seen a

live specimen—but Bill had, and his firsthand knowledge

helped him to laugh off my ill-informed speculations,

and keep our interpretations true to the reality of snake

ecology in southern Africa. Bill was a terrific collaborator

Amphib. Reptile Conserv.

Fig. 5. Jonno Webb, Bill Branch, and Peter Harlow posing at

the outskirts of the town of Poffadder in northern South Africa,

reveling in the idea that somebody actually named a town after

a snake (Photo: Rick Shine).

Pet i

and a wonderful friend. I feel privileged to have been

able to work with him.

James B. Murphy

Division of Amphibians & Reptiles, National Museum

of Natural History, Washington DC, USA

When we first met at a herpetological conference in the

US many years ago, Bill and I noticed that our love of

amphibians and reptiles, overall biological interests, and

personal histories were strikingly parallel. One major

difference was that Bill had completed a Ph.D. inchemistry

and I barely passed my chemistry courses. Fortunately,

he changed trajectories and excelled in herpetology. As

we shared our stories over some beers until the break

of dawn, Bill and I quickly bonded. I invited him to

come to Dallas, Texas, where I was herp curator at the

Dallas Zoo with a spacious guest room available in my

home. As we toured the Zoo’s herp collection, Bill was

delighted when he saw the large breeding group of New

Caledonian Geckos (Rhacodactylus leachianus, Fig. 6).

There was a particularly large and impressive male that

was surplus, so I gave it to Bill—his stunned reaction

and gratitude were wonderful to watch as he carefully

packed the saurian to hand-carry it back to Port Elizabeth

[ED note: This gecko is still alive in the Port Elizabeth

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Conradie et al.

Museum as of July 2019].

After the First World Congress of Herpetology in

Canterbury, UK, he invited me to stay at his parents’

home nearby until we later went to Bonn, Germany,

for the first Varanid Symposium held at the Museum

Alexander Koenig (Fig. 7). In my view, Bill was a

pretty stocky fellow, but his mother was concerned that

he was not paying enough attention to proper nutrition,

so she followed him for several days with handfuls of

vegetables, insisting all the while that he was becoming

a mere slip of a man. The scenario reminded me of a

Monty Python skit.

At the varanid meeting, Bill presented a wonderful

lecture on the White-throated Monitor (Varanus

albigularis)—“The Regenia_ registers of Brown

(1869-1909). Memoranda on a species of Monitor or

Varan.” Branch covered all aspects of Alfred ‘Gogga’

Brown’s extensive observations—sex ratio, size, body

proportions, hemipenial morphology, visceral fat bodies,

coloration, diet, cause of death, longevity, reproduction,

gestation period, egg laying, oviposition, eggs, clutch

size, hatchling size, incubation period, growth, behavior,

mating behavior, shedding, thermoregulation, predation,

parasites, exploitation, and seasonal activity and retreats.

The amount of information that Gogga had collected on

his captive lizards and in wild counterparts in the late

19th century is truly astounding.

Over time, Bill sent anumber of African and Namibian

reptiles for the Dallas Zoo collection, including Angulate

Tortoise (Chersina angulata), Parrot-beaked Tortoise

(Homopus_ areolatus), Tent Tortoise (Psammobates

tentorius), Mountain Adder (Bitis atropos), Dwarf Adder

(Bitis rubida), Many-horned Adder (Bitis cornuta), Cape

Dwarf Chameleon (Bradypodion pumilum), Lesser Flat

Lizard (Platysaurus guttatus), and Drakensberg Crag

Lizard (Pseudocordylus subviridis).

In the ensuing years, we spent much time together

at meetings and he shared his concern about shrinking

funding for the Port Elizabeth Snake Park and Museum.

Frederick William FitzSimons (born 1875) was the first

Director of the Museum in 1906, and he developed the

Fig. 6. Male New Caledonian Geckos (Rhacodactylus

leachianus) still alive in Port Elizabeth Snake Park (Photo:

Werner Conradie).

Amphib. Reptile Conserv.

vil

Fig. Z Varanid Symposium participants at Museum Alexander

Koenig in 1991. Bill Branch is the seventh person from the left,

in the front row.

Snake Park. His son, Vivian, assisted him and both of

them published in herpetology. His younger brother,

Desmond C. FitzSimons, started the Durban Snake Park.

F. W. FitzSimons also wrote books on the natural history

of South African mammals, including primates.

Bill was a consummate biologist whose contributions

to our knowledge of African amphibians and reptiles over

several decades set the high standard for herpetological

work. His nominal retirement as curator of herpetology

at the Port Elizabeth Museum occurred after many years

of service. As far as I know, he did not free-handle

venomous snakes nor put them on his head. Every time

we met, I could be confident that he would cover subjects

virtually unknown to me. He will be missed.

Luke Verburgt

Enviro-Insight & University of Pretoria, South Africa

Bill replied to my email almost instantly and in great

detail! I'd been very hesitant to contact my herpetological

idol about a reptile identification query, because I guess

I was afraid to disturb such an important person with

possibly silly and trivial queries from me, a nobody.

Yet to my delight, Bill took the time to carefully answer

my questions, providing great detail and assistance.

No admonishment for not having read the appropriate

books/papers, and no arrogant stance regarding my lack

of herping credentials! I was thrilled, and it opened up

communication between us to such an extent that soon

we were communicating about African herpetofauna via

email quite regularly, with Bill always helpful and kind

in dispensing his amazing wealth of knowledge. Like a

mentor really.

I eventually met Bill in person months later in

Namibia, along with Johan Marais and Aaron Bauer,

while they were on a collecting field trip. It was such

an honour to be sitting around the same table as these

herping heroes, and I was rather star-struck. After some

fieldwork with the team, I picked up on the fact that the

species of Rhoptropus that we were collecting was not

the one I had expected to be there according to Bill’s

field guide, which was really my main source of herp

September 2019 | Volume 13 | Number 2 | e186

Tributes to William Roy Branch (1946-2018)

knowledge, as it has also been for so many others.

I eventually plucked up the courage and cautiously

approached Bill one afternoon, to ask about this

Rhoptropus situation. He laughed heartily and said,

“Oh, that map is complete rubbish!" I couldn't believe it.

The author of the book that I cherished above all others

just told me that some of it wasn't the complete truth!

And right there I learned two massive lessons from Bill

about African herpetology: imperfect data from under-

sampling abound, and not being afraid to question the

existing understanding.

Sadly, that was the one and only time I got to be in

the field with Bill, and it was far too brief. Thankfully

though, we collaborated a lot after that with several

resulting papers where I got to be a co-author with

Bill—a huge honour! However, the greatest honour

for me in this regard was having Bill as a co-author

guiding my very first reptile species description, an

interaction through which I learnt more than I could've

ever imagined. He took what was a pretty ordinary and

mundane manuscript and guided me on how to improve

it to an acceptable standard, the standard which he was

instrumental in setting for African herpetology.

After that I regularly reported to Bill, who was always

extremely interested in my findings because I was often

working in poorly sampled rural places across Africa.

In his now familiar mentor role, he would encourage

me to do as much useful sampling as possible, and

also to think harder about why a particular species was

observed in the habitat I found it in and, therefore, to

consider its ecology in greater detail and gain more

insight from my observations. In short, Bill made me

a better herpetologist and I am forever grateful for his

friendship and his mentoring.

Although I didn’t see Bill in person very often, it was

always a treat to hang out with him and his fantastic

sense of humour. But what I think I enjoyed the most

was to hang out with him and to see him having such

fun at the 2017 HAA conference at Bonamanzi, and

I was even lucky enough to win a “selfie” with him!

Unfortunately, I never actually received the “selfie’—

but fortunately, Shivan Parusnath managed to capture

the “selfie”’-taking moment perfectly, and it is my

favourite photo of Bill and myself (Fig. 8).

I received the news that Bill had passed away while

I was sampling in the Cabinda Province of Angola, an

area of great interest for Bill. While we all had known

for some time that it was an inevitability, the news of

his passing came as a massive shock to me because

only a few hours earlier, during his last night, Bill had

somehow managed to send me a lengthy Whatsapp

message, instructing me to collect as many DNA samples

of certain species as I could due to the importance of the

sampling locality I was in. And thinking about it now,

that's just how it was always going to be for Bill, the

ever-enthusiastic herpetologist and helpful mentor to

the very, very end. Rest easy Bill, I miss you so much

Amphib. Reptile Conserv.

viii

AANA

Fig. 8. Taking a “selfie” with Bill as part of the prize for

runner-up best photographer (Photo: Shivan Parusnath).

and hope that I am able to justify the effort you put into

sharing your time and knowledge with me.

Julian Bayliss

Ecologist and Explorer, Wales

I first met Bill when he came to undertake a

herpetological survey on Mount Mulanje in Malawi with

Johan Marais and Michael Cunningham in 2005, as part

of the ongoing ecological monitoring programme on

Mulanje that I was coordinating. However, it was really

when we met the second time, when Bill and Werner

Conradie joined me on Mount Mabu in 2009, that we

really got to know each other well. I had been working

the mountains of northern Mozambique for several

years prior to this event, and had managed to turn up

several new species of snakes and chameleons, although

my herp work was opportunistic (I discovered Atheris

mabuensis by stepping on it!) and I needed professional

assistance (Fig. 9). These discoveries attracted Bill, and

we arranged for a trip to Mt. Mabu forest to collect more

specimens, and also to see if we could collect specimens

of a Nadzikambia chameleon that was only known from

a couple of photographs taken on my previous visits.

We were successful in this endeavour, and I managed

to collect the first specimen of the Nadzikambia from

Mt. Mabu which Bill named after me as Nadzikambia

Fig. 9. Photograph of Atheris mabuensis taken by Bill—

probably the best photograph of a snake I have ever seen

(Photo: Bill Branch).

September 2019 | Volume 13 | Number 2 | e186

Conradie et al.

Fig. 10. The Mt. Mabu 2009 science team. Left to right:

Werner Conradie, Martin Hassan, Julian Bayliss, Bill Branch,

Hassam Patel, Colin Congdon, and Steve Collins (Photo:

Julian Bayliss).

baylissi. | was deeply honoured by this gesture.

The 2009 Mt. Mabu expedition proved to be a

very enjoyable expedition, packed full of laughter,

good company, and good food. I had also invited the

butterfly crowd from the African Butterfly Research

Institute (ABRI), a great bunch of eccentrics, and the

stories flowed around the camp fires at night. At the end

of the expedition, we all stood below a large tree on

the forest camp in Mt. Mabu with Bill at centre stage

(Fig. 10). This is one of my favourite photos of Bill,

and it captures a moment in time where nothing outside

that camp at that time really mattered. This was the

start of a very good friendship with Bill (and Werner)

and some great correspondents. However, one of my

fondest memories of Bill was spending time with him

in the Mt. Nimba forest in Liberia. It was part of an EIA

on a proposed mining concession, and it was just the

two of us for several days, which gave us plenty of time

for chewing the fat; especially when we talked about

rugby and Wales vs. South Africa or England, as I am

from Wales and Bill was originally from England, and

then South Africa. At that time, I had flown up from a

festival in South Africa and brought with me a ‘Green

Policemen’ helmet which Bill dually wore (Fig. 10, this

photograph shows Bill beaming a big smile).

Bill, I will miss you greatly—you were an inspiration

to me. Not only did you teach me a lot about reptiles, but

you were also a professional in everything else you did.

An expert and a gentleman. In the last communication I

received from Bill, a couple of months before he passed,

he told me ‘not to defer my dreams’—advice which is

applicable to us all and advice I intend to follow.

Michael F. Bates

Department of Herpetology,

Bloemfontein, South Africa

I knew about Bill soon after I started working at the

National Museum in Bloemfontein in 1983, as he was

then editor of the Herpetological Association of Africa’s

journal. The first time I met him was at the HAA’s first

National Museum,

Amphib. Reptile Conserv.

Fig. 11. Bill Branch in the Nimba forests close to Nimba

Mountain, Liberia (November 2011). Bill is wearing the

green policeman hat I had brought with me from South Africa

(Photo: Julian Bayliss).

conference held at Stellenbosch University in 1987. I

was only 25 at the time, and Bill was about 41, still quite

slim and with a full head of black hair! At that time he

was busy wrapping up work on the first edition of his

famous reptile field guide. Even then I remember Bill

having a certain charm about him and the aura of a man

with a deep knowledge of his subject matter.

Over the years I visited Port Elizabeth Museum

several times to examine specimens for various research

projects, including some on which I collaborated with

Bill. Having him all to myself and available to answer

my barrage of questions was always special. However,

I think my fondest memories were in the early 2010s

when we spent considerable amounts of time editing the

text for the Atlas and Red List of the Reptiles of South

Africa, Lesotho and Swaziland (published in 2014). As

first and second (Bill) editors, the bulk of the editing

fell on us. I would, for example, e-mail Bill the text

for a species account and ask such questions as “‘is it

still regarded as a subspecies” or “has anything been

published about this recently.” I could count on him to

respond within a day or two, and his responses were

always insightful. He seemed always to be up-to-date

with the latest taxonomy and the most recent literature.

And so it was that we e-mailed the various sections of

text back-and-forth until we were both happy. I have

very good memories of those times.

Another special memory I have of Bill was in May

2018, a few months after he was diagnosed with MND,

when I visited him at home in Port Elizabeth, together

with Aaron Bauer and Marius Burger. By this time he

was, for the most part, wheelchair-bound. Nevertheless,

he was as talkative and interesting as ever, especially with

regard to herpetological matters, and he also exhibited

his usual great sense of humour. We spent most of the

time at the computer in his study where he showed us

photographs of interesting and new reptiles, and of field

trips he had conducted with various colleagues over the

years. Also, I brought him a copy of a recent taxonomic

paper on egg-eating snakes (Bates & Broadley) that

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Tributes to William Roy Branch (1946-2018)

Fig. 12. Michael Bates (left) with Bill Branch and Darren

Pietersen during the Herpetological Association of Africa’s

conference in Pretoria in 2013 (Photo: W.R. Schmidt).

had just been published in the National Museum’s

journal Indago. The front cover of the journal featured a

montage of Bill’s excellent colour photographs of these

snakes, and it gave me great pleasure to see how pleased

he was with the way it turned out.

Bill had an enormous presence in the field of African

herpetology. He impressed me as a very well-read

man, and this was reflected in his wide and seemingly

limitless knowledge of reptiles and amphibians. Bill was

always willing to share PDFs of research articles and in

this way he helped me on innumerable occasions. Also,

I was inspired by his style of writing and attention to

detail. I still think about Bill often and will miss him for

several reasons, not least for the fact that his expertise

was always just an e-mail away.

Pedro Vaz Pinto

Kissama Foundation, Luanda, Angola & CIBIO-

InBIO, University of Porto, Portugal

I first met Bill in January 2009 in the most appropriate

of places: deep in the Angolan Namib desert, in

Iona National Park. We were part of a large group of

scientists assembled by Brian Huntley for a biodiversity

expedition in southern Angola. I remember approaching

Bill after dinner in the camp site, and he was keen to

see my photo files and became interested in some bush

viper pictures, which led to a few engaging stories and

discussions. At that point I was simply curious about

reptiles, and more involved with furry or feathered

creatures. The following day, I drove my Land Cruiser

to where I could see Bill and his colleagues had parked

their pick-up truck next to some granite boulders. I could

sense some excitement in the party, so I asked Bill what

they were doing. He invited me to join them and opened

a little box to retrieve a tiny beautiful little gecko, one

of the gems of Angolan herpetology which was not even

formally described at the time: the endemic Plume-

tailed Gecko, Kolekanus plumicaudus! He then showed

what was special and unique about that species and

chatted about other leaf-toed geckos. I was fascinated of

course, and it was quite an introduction to reptiles. Over

Amphib. Reptile Conserv.

Fig. 13. Bill Branch processing specimens in the fading light

of the Angolan Koakoveld (Photo: Pedro Vaz Pinto).

the following years we would become good friends, but

looking back I’m still amazed to realize how generous

he was by sharing that amazing find with someone he

had just met the previous day. Other scientists would

have kept their cards very close to the chest. But Bill

kindly drew me towards the world of herpetology for

which I’m forever indebted, but above all I believe

he made me a better scientist and better naturalist. He

taught me to make an effort at looking into the bigger

picture, to see the multiple layers and connections that

lie hidden behind the outer surface of a given ecological

theme.

My best memories with Bill, without any shadow of

doubt, were the days in which I was privileged enough

to travel with him to some of the most remote corners,

wildest places, and biodiversity hotspots in Angola.

We would typically look for a scenic landscape off the

beaten track and choose our camping spot. Some of

the time shared with Bill, around the campfire in the

Angolan desert, mountains, or forests, was memorable.

Our camping expeditions were hugely stimulating

scientifically, exciting and unpredictable, and very

importantly, always bathed by loads of good humor!

These expeditions could be physically exhausting, but

soon after I was looking forward for the next trip with

Bill.

Other scientists are much better prepared to praise

Bill’s unique and extraordinary legacy to African

herpetology. I can add that he did leave a crucial mark on

Angolan herpetology, but tragically with his premature

passing away, it wasn’t allowed to further crystalize

during his life. He was arguably the most influential

herpetologist to have worked in Angola for a sustained

period, and is the main person responsible for bringing

herpetology into the biodiversity agenda in modern

Angola. I have no doubt that his pioneering role will be

recognized in the future by young Angolan biologists.

On a personal note, whenever we came across a new

lizard or snake, I got used to my sons asking me ‘- Will

Bill want this specimen?’, ‘- Has Bill identified this

species?’, ‘- Does Bill need more specimens?’ and as

result, these now rhetorical questions remain quite vivid

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Conradie et al.

Set

ge |

Sy S \ : —— oh. Sa age S-

Fig. 14. Bill Branch photographing a Jameson Mamba

Angola with Ninda Baptista (Photo: Pedro Vaz Pinto).

and still drive me on my searches. There are still a lot

of ‘goodies’ that we will catch for you Bill, and that’s

a promise!

Kirsty Kyle

KwaZulu-Natal, South Africa

I had the good fortune of growing up at Kosi Bay

with Bill Branch as a much loved family friend. Bill

had gotten to know my parents, who were the resident

scientist and his wife for the area. In those days, he did

an almost annual foray to Zululand, what with it being

such an interesting part of the country for herpetofauna.

Whenever he moved through the area, with his pack

of scientists, they would use our house as a base, and

for my two older brothers and I this was just the best

thing ever. His trips became the highlight of our year

and I think he thoroughly enjoyed having three young,

able-bodied slaves, ever so willing to dive after any

reptile that was silly enough to stick its nose out 1n our

vicinity. A friendly disagreement developed as we got

older and started objecting to his pickling tendencies.

In the later trips we would “not see” a lot of the more

common species because dear old Uncle Bill would just

pickle anything we presented him with, which was a bit

hard on our budding conservationist hearts. Although

we had a pretty much genetic interest in herps, I think

those times with Bill were extremely formative in all

three of our lives, they certainly were in mine. The fact

that he was interested and enthusiastic in teaching and

encouraging a little blonde thug of three years old in the

ways of reptiles was amazing.

Bill was absolutely instrumental in setting me on

the path I am on today. Throughout childhood it was

a privilege to spend time in the field with him and just

absorb all the information he so generously and freely

dished out. It had a major impact on my interest in

herps. I emailed back and forth with him whenever I

found something interesting, and I sent him pictures of

all sorts of different reptiles over the years and he would

always respond in his warm, friendly, and encouraging

manner, which was just amazing. My favourite memory

of Bill would have to be on his last visit a few years back,

Amphib. Reptile Conserv.

when he proudly presented us with a beautiful pot that

Donvé had made, decorated with an aloe he informed

us he’d just plucked from our outdoor lizard enclosure.

There were no flies on Bill and I loved that about him,

he always told you the truth, even if it put him in not the

best of lights. We still have the aloe in the pot.

I wish I had a picture with him from the early days

because it really would be a cute one. I fondly remember

parking on his lap as a very little girl, discussing

whatever, feeling terribly important, with his black

mop of curls and my blonde mop of curls. It would

have been such a cute picture. I miss Uncle Bill, the

world in general is a lot less fun without him and my

Facebook is a much darker place without his frequent

updates, pictures, thought processes, and quips. I hope

he forgives me for specialising in amphibians instead of

reptiles, and I’m incredibly grateful to have had him as

a friend, as well as a mentor.

Krystal Tolley

South African National Biodiversity Institute, South

Africa

I knew about Bill before I moved to South Africa in

2001, as he and Colin Tilbury had some chameleon

DNA samples for my upcoming postdoc project. The

project almost didn’t happen, as Bill and Colin got cold

feet, but when I arrived I learned that they decided to

let me give it a try. Their trust in a stranger with whom

they had never worked ended up building a friendship

and collaboration that lasted nearly two decades. As

that project progressed and more projects arose, Bill

encouraged and supported me both in a personal and a

professional capacity. In fact, the entire herpetological

community welcomed me, something that I was not used

to, coming from the competitive world of marine biology

in the northern hemisphere. Because of Bill, Colin and

all the SA herpers, I felt like I had found a home that I

didn’t want to leave, and Bill was instrumental in that.

I cannot remember actually meeting Bill for the first

time. My first distinct memories of hanging out with

Bill and all the herpers is from the Port Elizabeth HAA

conference in 2004. What sticks out in my mind 1s that

at the concluding banquet, Bill received the Exceptional

Contribution to African Herpetology Award and he was

so touched by this that he wept. That spoke to his nature

as a caring person who knew that strength and courage,

not weakness, comes from personal relationships

and bonds. And his connections with his friends are

something he fostered.

I have many distinct and fond memories of Bill but

strangely enough, most of them relate to our friendship,

not to herpetology. When we would meet, the first

things he would ask me about was how I was, how was

my personal life, what was happening, was I happy?

He had many wise words for me along those lines,

giving advice, encouragement, and reassurances that

eventually I would find my path. Then of course, the

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Tributes to William Roy Branch (1946-2018)

‘herp talking’ would start. He would go on for hours,

non-stop, about snakes mainly. Most times, the topics

were just beyond me. I tried to absorb what he said, but

there was so much information that my brain couldn’t

handle it. I do remember that a long discussion about

Leptotyphlops made me realise what cool things they

are, and I still hope one day to actually work on them.

I was fortunate to have the chance to visit Bill shortly

before he passed away. We both knew, as did his wife

Donveée, that I was there to say a final goodbye. This was

indeed the last time I saw him and it was emotional for

everyone, but my memory is still a good one. The same

old routine was there. He asked me about my life first,

and he gave me wise words and insight about life. Then

he spoke about herps (including Leptotyphlops) for

about four hours non-stop. The thing that was different

this time, was that he often interjected the conversation

with things about himself. Dreams and wishes, failures,

successes, lost opportunities. He talked a lot about how

it’s important not to waste time on petty or destructive

things in life. But to focus time and energy on the people

in your life that care about you and to never take that for

granted. He spoke about the balance between the work

related passions of a herpetologist, and that this has to

balance with life, friends, and family. Bill was a hard

worker, but he did focus on family and friends, and I don’t

think he took any of that for granted. The way that his

first questions always related to our personal connection

and friendship, and about which analysis I was running,

speaks to that. The wisest words that Bill ever said to me

are: “Friendship 1s a gift. It’s a gift that others chose to

give, and that you chose to accept in whatever form it

takes.” Bill gave that gift to me and to so many others.

That is what I will remember him for the most.

Fig. 15. Bill Branch and Krystal Tolley in south-western

Angola in 2009 (Photo: Krystal Tolley).

Mzi Mahola

South Africa

I first met Bill when he arrived at the Museum. A year

or two later, I invited him to join our Port Elizabeth Mu-

seum soccer team, which was playing in the Industrial

League. He didn’t play many matches, because of his

Amphib. Reptile Conserv.

xii

commitments, but he was a good soccer player. A year

later, I was transferred from my department to join and

work with him as his research assistant. We often went

to Sardinia Downs to tag, study, and monitor the move-

ments, growth, and development of tortoises. After that

he introduced me into his other research and study pro-

gramme of other animals, such as frogs, snakes, and liz-

ards.

One day we were going up the Zuurberg Mountains

when he made a deal for us; “If you happen to catch a

snake first, I will buy you a bottle of beer at the Zuur-

berg Inn on the way back; but if I make the first catch,

you will buy me a bottle.” That was fair enough for me.

Bill was at the wheel of our Land Rover. We were driv-

ing towards the forest at the foot of the mountain when

I saw a female boomslang flying towards the forest. Bill

noticed my hasty intention to open the door of the mov-

ing vehicle and he quietly said, “Forget it! Boomslangs

are very shy; you'll never catch it unless it is ina tree.” A

few minutes afterwards, we left the Land Rover and with

our hunting gear and went our separate ways. It took me

less than five minutes before I heard the hissing sound of

a slithering snake. I saw the disappearing tail of a rinkals

entering a hole amongst rocks on a ledge. I put on my

safety glasses and peeped into the hole, and saw the two

shiny eyes watching the entrance. With my tools I pulled

the snake out and put it in my canvas bag, and declared

my victory to Bill. He didn’t believe me. It didn’t matter

how many snakes he collected afterwards. I had beaten

my master in his game and the bottle of beer would be a

cherry on top.

We were on a trip to the Drakensburg Mountains and

our first night stopover was in Centane, at my in-laws. We

shared the same bedroom. At night, Bill said something,

which I could not let pass unchallenged; “Kentani is the

only place, in the Eastern Cape, that has no tortoises.”

“Why?” I asked, thinking that this had to do with the

climatic environment.

“Africans ate all of them and left nothing to sustain

these animals.”

“No! That is not true!” I protested, because I had a

relationship with these people.

“What do you mean, it is not true? Dr.... (/ dont re-

member his name) learnt about this when he was inves-

tigating the cause of their depletion in this area, years

back in the early twenties. I read his book and you can’t

dispute it.”

“Well, his assumption was wrong.” I replied, confi-

dently, knowing that what he was going to hear would

shock him. “First of all; it 1s very, very difficult for a

stranger to get information from traditional amaXhosa,

because these people are known for their scepticism of

strangers. If you ask them anything, they will ask, “why

do you want to know?’ After that they will not share with

you their knowledge; more so if you’re a stranger. In

the past amaXhosa trained their children from an early

age never to tell a stranger the truth, especially to white

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people.”

Bill listened quietly, without interrupting.

I continued, “Now let me tell you something that they

did not tell Dr.....? AmaXhosa did not eat any creeping

or crawling animals, like centipedes, lizards, snakes,

crabs, frogs, locusts, ants or tortoises, hence they looked

down upon Khoisan people, because of their “repulsive”

diet. The Khoisans ate these animals.

Even though they were converted into Christianity,

there are still some Xhosa households who hunt and kill

or keep tortoises for their strong religious or cultural be-

liefs. They generally believe that if they burn a tortoise

Shell in a kraal with cattle, the cattle will multiply. Cat-

tle are a status symbol or a bank to our people. Tortoise

shells are also used as troughs to store drinking water for

chickens so that they may increase. There is also a belief

that if live tortoises are kept in a household, they will

repel evil spirits. These beliefs surely must have been the

cause of depletion of these animals in an area as conser-

vative and traditional as Centane. I was told that because

of their scarcity, locals are prepared to purchase and im-

port them from other areas.”

“It makes sense,” Bill said and kept quiet for a long

time afterwards.

Working with Bill had a very strong impact on me. He

was very dedicated and committed in whatever he was

doing. In Matatiele, he went out into the night to search

for frogs in the river while it was raining and thundering.

He didn’t allow anything to stand in his way. Many years

later, after I had left P. E. [Port Elizabeth] Museum, I

went on a personal and voluntary excursion of document-

ing and taking pictures of bushmen paintings in the caves

in the Nkonkobe and Chris Hani Municipalities. Without

his basic research training I wouldn’t have embarked on

this project. Bill gave me a hands-on experience in re-

searching and I thank him for sharing his skill with me.

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Fig. 16. Bill Branch discussing the finer points of the day’s

photographic record of collections with colleagues and

students, Lagoa Carumbo, May 2012. (Photo: Brian Huntley).

Amphib. Reptile Conserv.

Brian J. Huntley

South Africa

During 2009 and 2012 I had the pleasure of introducing

Bill Branch and a few dozen other field biologists to

the diversity of life in the deserts, montane grasslands,

miombo woodlands, forests, and floodplains of the far

reaches of Angola. Bill soon proved to be the hardest

working and most convivial member of the teams, which

comprised up to 30 biologists from ten countries.

During the first expedition to southwestern Angola,

in January 2009, we camped out on the Humpata

highlands and in the Namib desert. Here Bill and fellow

herpetologists found multiple new records and several

new species of frogs and lizards. From the faint light

of dawn to the pitch darkness of night Bill would be

in the field or at the makeshift laboratory tent, where a

generous donation of Cuca beer from the local brewery

kept spirits and laughter levels high. What impressed me

most about Bill was his ability to inspire all around him—

young students to ageing professors—game rangers to

army generals—with fascinating stories about his cold-

blooded friends.

In May 2012, when we were camped out along a

Congo tributary at Lagoa Carumbo, in the far northeast

of Angola, the evening’s discussions around the campfire

ranged from Bill’s erudite interpretation of current species

concepts to scary personal experiences of snake bites and

the treatment thereof. We were eight hours drive from

the closest town, and another five hours from the closest

doctor. One afternoon, Bill had been out to set a trap for

a black mamba that had been seen slithering down a hole

on arock face. He casually told us how he had, that same

morning, pulled what he thought was a harmless water

snake out of the Luele River. Only when he returned to

camp did he discover that it was a new elapid record for

Angola — Banded Water Cobra, Naja annulata.

We had planned to visit Portugal together early in 2018

to discuss collaborative projects with colleagues at the

University of Porto, but at a meeting in Cape Town that

January, Bill informed me of the advice his doctor had

given him that week: he should not travel. We soon learnt

of the severity of his illness, but this did not slow Bill

down. He was already under heavy pressure to complete

his catalogue of Snakes of Angola, but did not hesitate

to honour his promise of a chapter on reptiles for the

synthesis volume that I was coordinating on Biodiversity

of Angola. We kept up a lively correspondence to the

end, his sharp wit never failing. Fittingly, given Bill’s

tremendous role in inspiring young researchers in Angola,

the synthesis volume includes a dedication to him.

Roger Bills

South African Institute for Aquatic Biodiversity, South

Africa

I first met Bill in Marromeu, central Mozambique. We

were part of a team lead by Jonathan Timberlake looking

at the biota of the lower Zambezi’s delta region. I had

September 2019 | Volume 13 | Number 2 | e186

Tributes to William Roy Branch (1946-2018)

driven up from Grahamstown with a bakkie and trailer,

and had slept in the car over several nights due to poor

road conditions and slow progress. Basically, I was

exhausted. This did not get me any respite from Bill’s

sharp humour and I had to quickly shape up.

For me the trip was tremendous—there was water

everywhere despite it being the dry season, and an

abundance of fishes and the fauna was mostly new to

me. For most of the other zoologists 1t was not the best

season and consequently a bit frustrating. Bill and I got

on from the start and we spent time out in the dry fields

with a large bulldozer that was flattening termite mounds

looking for snakes and lungfishes and several days on a

boat going down the Zambezi.

The boat trip down the Zambezi was supposed to be

an overnight affair—down the Zambezi to the mouth, a

channel through mangroves to one of the delta’s southern

braids and up to the small village of Malingapanzi.

Unfortunately we missed the tide and left late, and went

down river on an incoming tide. It took us the whole

day to get down to the mouth where we camped at a

fishing village overnight. We expected to get going at

first light but the local fishermen stole our rudder as they

wanted payment for camping. It took our Mozambique

counterparts the whole morning of negotiating and

refitting the rudder before we could leave. The time

however was well spent: Bill went fishing (he was a good

angler) and caught our only Glossogobius giuris for the

trip, and I caught a load of mud-skippers in the mangrove

flats. Our delay meant we missed the tide again and going

up the southern channel to Malingapanzi was against the

outgoing tide. We got there late on the second day—Bill

had caught one puff adder. He wasn’t very happy and did

not return by boat the following day.

From all my experiences with Bill, the impressive

thing about him was his resourcefulness in the field,

whether collecting by himself or soliciting samples from

locals, he managed to get incredible numbers of samples.

Returning to camps in the evenings would invariably find

Bill at a table covered with specimens that he would be

fixing, photographing and taking tissue samples from. He

Fig. 17. Bill and Anton Bok at the Kalumbila Mine Camp,

Mwinilunga District, North-West Province, Zambia, May 2010

(Photo: Roger Bills).

Amphib. Reptile Conserv.

spent long hours doing this work. On one trip to a sand

mining project near Pebane, Mozambique, we fell afoul

of this. Bill had been there the week before and the locals

were used to giving reptiles they had caught to passing

vehicles. On our drive from the airstrip to the exploration

camp we were oblivious of this. After the second snake

came through the window in a flimsy plastic bag, we

wound up our windows and did not stop anymore!

Bill was an incredible intellect, a world-class scientist

but far more importantly a great guy. It was a privilege to

have spent time with him, my life 1s richer for it.

Johan Marais

African Snakebite Institute, South Africa

Back in 1980, while I was curator of Transvaal Snake

Park, I met Bill during one of his visits but we barely

spoke. I was a youngster cleaning snake cages and Bill

was visiting Rod Patterson and Anthony Bannister. We

often corresponded and I supplied Bill with a bunch of

photographs for his field guide, but it was only in early

2000, on a field trip to Namibia, that we really bonded.

We did several field trips to Namibia, often with Aaron

Bauer, but our trips to Niassa in northern Mozambique,

Mulanje Mountain in Malawi, and southern Angola

were memorable. Field trips are special as there 1s ample

time to chat, especially when driving long distances. I

particularly enjoyed the chats with both Aaron and Bill,

and although there were endless topics discussed it

was largely about reptiles. I often wound Bill up about

photographing reptiles on inappropriate props like fruit

and flowers that were out of place, and he accused me of

taking rather poor photographs as I had a bad eye.

His wry sense of humour brightened things up on

those long journeys and he was particularly good at

irritating Aaron, not to mention times when he would

lose specimens while photographing them! My best

Bill moment: when an American missionary’s wife

in Nampula asked Bill what he does for a living, he

responded that he was a reptile scientist who did field

work, described recently discovered reptiles, and wrote

scientific papers about his discoveries. She responded:

i Po, .

i —

Fig. 18. Johan Marais and Bill Branch with a Rock Monitor in

Namibia. (Photo: Jackie Childers).

September 2019 | Volume 13 | Number 2 | e186

Conradie et al.

“Yes but what is your real job?’

It is hard to grasp the gap that Bill has left behind, and

so many of us miss the times that we could call or drop

him an E-mail. He was notoriously bad at responding to

E-mails so I got into the habit of numbering my questions.

Needless to say, Bill would only answer those he felt like

answering.

Mark-Oliver Rédel

Museum fiir Naturkunde, Germany

My first contact with Bill was in 1996. He asked for a

copy of my frog book, and invited me to give a talk on

West African amphibians and reptiles on the third World

Congress of Herpetology in Prague, where I met him for

the first time in person. Bill was organizing a session to

summarize the progress in African herpetology. Thus, it

was Bill ‘officially’ introducing me, my Ph.D. not yet

finished, to the community of African herpetologists. We

kept contact thereafter, but it took a few years until we

met again.

Following a workshop to define conservation

priorities for West Africa, Conservation International

started a series of rapid biodiversity assessments in little

known areas across the Upper Guinea forests. In early

2002, Bill and I were asked to participate on one of these

RAPs, targeting the Haute Dodo and Cavally Forest

reserves in western Céte d’Ivoire. He was responsible

for the reptiles and I was to focus on amphibians, but

of course we conducted all field work together, recorded

many interesting amphibian and reptile species, ignored

all CI safety rules, and had a lot of fun catching animals

and talking rubbish. For Bill it was his first time being

in West Africa, and his first time working in rainforests

(as a ‘typical’ South African he showed up in shorts and

it took me quite a bit to convince him that working in a

rainforest in shorts is a very stupid idea).

Not all of the experiences were fun. In one night in the

Cavally forest, we walked far from camp and encountered

a few rarer species we hadn’t seen before on the trip. On

our way back, we stumbled straight into the largest raid

of army ants (Dorylus sp.) I ever encountered! The forest

floor and all lower parts of the shrubs and trees were

covered with these aggressive insects, and in seconds the

ants where everywhere on and under our clothes. We just

ran to leave them behind, and then had to strip naked to

pull off hundreds of ants, all holding onto the skin they

had successfully penetrated with their sharp mandibles. It

was only when we finally finished them all off (one has to

pinch off the heads of every single one) and turned again

towards the campsite, that Bill realized that he had lost

his glasses. We had to turn back into the ants to search for

them..... A much more pleasant experience on that trip

was when we found the first live caecilian, Geotrypetes

seraphini, Bill had ever seen.

Caecilians were also one of the most spectacular

findings, actually the first country record for the entire

group, the next time we met. In the fall of 2003, Bill

Amphib. Reptile Conserv.

invited Johan Marais and I to survey amphibians

and reptiles in the Niassa Game Reserve, in northern

Mozambique. Although it was the core dry season it was

an extremely successful survey, revealing 57 reptile and

31 amphibian species, including a new Cordylus, and

further potentially undescribed species including the

Scolecomorphus mentioned above.

Thereafter we met regularly, mostly in South Africa,

but a few times in Germany as well. Bill often took me on

shorter excursions across southern Africa, e.g., showing

me spectacular parts of the Cape Fold Mountains or the

Karoo, and I frequently visited him and his wife Donvé

in their amazing house and garden in Port Elisabeth.

There we had long and entertaining discussions about

herpetology, science, politics, or sports, while sipping

on a nice glass of wine, observing the many birds in the

garden, or following a soccer or rugby match on television.

We never agreed on which soccer team or player was

worth supporting, and I could always bet that I would

receive a derisive email after a German defeat against

an English team in the Champions League. Bill was

mad about some sports and missing an important rugby

match was impossible, even on an excursion. Particularly

memorable was when we once drove through the Karoo

and he wanted to listen to a match on the radio. As the

radio quality was weak, we had to finally stop and follow

the broadcasting on the roadside in the desert. However,

the only program Bill could find was in Afrikaans. Thus

apart from the players’ names and the score, he did not

understand a single word. An amazing fact about Bill

was that, although he was a forceful speaker, loving

to use and to play with the English language, he was

completely ignorant about other languages. So he never

learned Afrikaans and in other countries, other people

had to cope with translations.

But Bill had encyclopedic knowledge of the natural

sciences in general, and he could instantly give a lecture

about southern African zoology, botany, or geology.

He was easily connecting all this different knowledge

into a broader, comprehensive framework and thereby

developing new questions and ideas. This ability to

communicate new or complex knowledge made him a

very stimulating academic teacher, something which was

certainly was one of the reasons why he was so popular

on the National Geographic tours he was guiding in his

later years. His non-protective way of openly sharing data

and ideas, as well as critically and without any mercy

dissecting project ideas, hopefully remains a model

to all the many students and colleagues with whom he

was communicating his entire life. Many of his ideas

and projects now remain to be finished by others, most

prominently the revision of the ‘bible’ Bill Branch’s Field

Guide to Snakes and other Reptiles of Southern Africa,

and the description of dozens of new reptile species he

had already collected and deposited in the herpetological

collection of the Port Elizabeth museum.

To me, Bill was much more than a good colleague,

September 2019 | Volume 13 | Number 2 | e186

Tributes to William Roy Branch (1946-2018)

Fig. 19. Bill Branch in a sad mood after his snake stick,

proudly stolen from Aaron Bauer, broke while he tried to

destroy an Opuntia in the Karoo, October 2012. Bill: “What an

embarrassing death to a snake stick, killed by a plant” (Photo:

Mark-Oliver Rédel).

although we did only meet occasionally. More often

in recent years, he was a very good friend with whom

I enjoyed discussing everything, not only science.

However, the scientific discussions with him were a

constant inspiration providing me with many, sometimes

unusual ideas on how to interpret data or set up new

projects. He introduced me to the African family of

herpetologists and to Mozambique; and I am proud that

I could introduce him to the West African herpetofauna

and rainforests, and even convinced him (sometimes)

that amphibians are not completely boring. For him, I

would have even loved to see England take the World

Cup in 2018. He died too early and in an unbelievably

Fig. 21. Bill Benen a Mark- Oliver Rodel wan the halite

of the species named in their honour (Photo: Frank Tillack).

Amphib. Reptile Conserv.

xvi

& |

¥,

Fig. 20. Bill Branch and Mark-Oliver Rédel in July 2018 in

Bill’s home in Port Elizabeth (Photo: Mark-Oliver Rédel).

cruel way. I am very happy that I could meet him one

last time, shortly before his death in PE. He will always

remain an unforgettable person and inspiration. His death

is a great personal and scientific loss, and my thoughts

are with his beloved wife Donvé and both their families.

Paul H. Skelton

Wild Bird Trust, National Geographic Okavango

Wilderness Project

When the National Geographic Okavango Wilderness

Project (NGOWP) was looking for key specialists to join

them on expeditions into the unexplored highlands of

Angola, Dr. Bill Branch was a first port of call. Bill was

attracted into the NGOWP as an established authority

of Southern African and Angolan herpetology, most

especially the reptiles. He joined the founding 2015

NGOWP Expedition as part of the 'land party.' He also

took part in the 2016 expedition, joining it after first

enjoying an extensive journey through the escarpment

reaches of Angola in the company of Dr. Pedro Vaz

Pinto. Prior to this, he had visited Angola on a number

of occasions, collecting and adding significantly to the

herpetofaunal knowledge of the country. His collecting

antics often drew curious onlookers, mostly children, who

would marvel at what wonders he would bring forth from

the ponds, rocks, and crevices. More significantly he both

encouraged and actively mentored younger hepetologists

currently active in Angola. These expeditions have

resulted in several potential new species, a number of

new species for Angola, and range extensions of many

others.

Bill was an old friend and colleague of some of us.

I personally met and knew Bill soon after he arrived in

South Africa, and was working for the Atomic Energy

Corporation outside Pretoria. On joiming the Port

Elizabeth Museum, we became good friends and he

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Conradie et al.

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Paullin).

joined me on at least two interesting expeditions that I

arranged primarily for fish sampling—one was along

the Lower Orange River, and the other was to Lesotho

in 1988. Bill had a'nose' for reptiles. I remember vividly

how he would relish the refreshment breaks on the

expedition as we travelled north across the karoo, in

order to sniff around the rocky kopjes and outcrops along

the way. He inevitably returned with a clutch of reptiles

in hand, all of which were unerringly shot with an elastic

band from the hand. And then there was the spitting

cobra, Naja nigricollis woodii, he caught by the tail on

crossing the road, after screeching to a halt and leaping

forth from the vehicle. As viewed from the vehicle,

behind it was an energetic spectacle in madness—born

out on arrival to realise that Bill had been spat in the eyes

by the enraged reptile and, whilst he was blinded and in

agony, was desperately directing his non-herpetological

colleagues in the niceties of bringing a canvass bag to

bear so he could insert the writhing beast. Needless to

say, he succeeded and managed to wash his eyes out

before he was permanently damaged. On the Lesotho

trip, Bill was his amazing self and not only displayed

his fly-fishing skills that I never knew he had, but also

showed me the cryptic, super-jawed, Maluti River Frog

(Amieta vertebralis) in its natural habitat. His calm

demeanour, bubbling humour, and all-round knowledge

in the field was always refreshing. Simply put, Bill was

a pleasure to have around. His scientific productivity and

achievements are of a top order. His passing was a great

loss to our project and to the community at large.

Aaron M. Bauer

Villanova University, USA

I met Bill in 1987, during my first trip to South Africa.

I had met with Alan Channing in San Francisco and he

had given me a list of all of the critical herpetologists and

institutions to visit in South Africa. After visits to Wulf

Haacke at the then Transvaal Museum and to FitzSimons

Snake Park in Durban, I made my way to Port Elizabeth

via Cradock. I phoned Bill on the way (from a post

office, remember no cell phones?) and he suggested that

my field assistant and I stay at the camping ground on

Amphib. Reptile Conserv.

Fig, 23. Bill Branch in Angola bean for ean: (Photo:

Alex Paullin).

Brookes Hill. A gale was blowing and we were soaked

to the bone, but the next morning Bill kindly showed

me around the Museum complex. Over the next day or

two he took me Bradypodion hunting in Happy Valley,

just down Beach Road from the Museum, showed me

the introduced Lygodactylus capensis on the guard rails

along the roads, and sent me off to Schoenmakerskop to

look for Acontias meleagris, Homoroselaps lacteus, and

other reptiles. Like everyone I met on that first trip, Bill

was a critical contact if I was intending to start working in

South Africa. By 1989, I was coming regularly, sometime

Fig. 24, Bill Branch in Angola with a dead on the road Vine

Snake (Photo: Alex Paullin).

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Tributes to William Roy Branch (1946-2018)

Fig. 25. “Uncle Bill” enjoying the adoration of the masses

(Marius Burger and Krystal Tolley) at the H.A.A. meeting in

Cape Town in 2011 (Photo: Aaron Bauer).

two or three times a year, and more often than not Bill

and I would go to the field together, starting a 30 year

personal and professional collaboration that influenced

all of my work in Africa as a whole.

I have very many fond memories of Bill. One was a

1990 trip to northern Namibia. I picked up Bill and drove

with him and two of my students to a farm in Kamanjab.

We stayed with the farm managers and had a wonderful

time. The collecting was spectacular and mostly new

to both me and Bill, who had not spent much time in

Namibia before this. Every day we found additional

species, in the end nearly 50 species on the farm alone,

and more between Kamanjab and Palmwag. Bill had

to leave before me and on his last night, after weeks

of the best warthog and gemsbok, we were promised

“something special,” which turned out to be a very old

and very gamey goat! The next day Bill and I left the

students and drove straight through to P.E. with only a

short stop for a nap. Our only music in the car was The

Greatest Hits of Elton John. On the trip we really got to

know one another, and we both got so sick of Elton John

that we couldn’t listen to his music for years.

Other fond memories are of our multi-year projects

in the Little Karoo and later the Richtersveld. In those

days there were very few visitors to the Richtersveld,

and Bill and I both enjoyed the solitude of the park,

evenings by the fire along the Orange River, and finding

two Bitis xeropaga only meters away from one another.

I can also mention a magical trip to the Kaokoveld along

with Johan Marais and my Villanova colleague, Todd

Jackman. We were in the bed of the Munutum River and

all of a sudden we were surrounded by a herd of giraffe.

Even Bill, always ready for the good photo opportunity,

was temporarily awestruck by the scene. I also spent

many memorable weeks with Bill in the States. One trip

was to the South Carolina coast just after a hurricane.

Despite some serious close calls with disaster, the loss

of one of Bill’s cameras, and hundreds of mosquito bites,

Bill was pleased to catch a baby alligator and to have

Amphib. Reptile Conserv.

had the chance to be in the field with Whit Gibbons, a

great herpetologist and ecologist, and an author whose

writings Bill admired. On another trip, we drove 10,000

km from coast to coast and back in the US with my

students and postdocs. At 3,700 m we saw a herd of

elk and Bill managed to get most of his body outside

of our moving van to get the perfect shot. I think all of

these fond memories are united by the common theme

of sharing with Bill the feeling of how lucky we are to

have a vocation we love and that lets us enjoy spectacular

animals in amazing places in the company of our friends.

Bill was the face of South African herpetology, indeed

of African herpetology. His interests were wide-ranging

and he had a mind for details when it came to all things

herpetological. He was also a master naturalist who knew

his birds and his plants, as well as the history of natural

history exploration in Africa. He was also down-to-earth.

Even the most novice of herpetologists was welcome to

call him Bill, not Dr. Branch. Although he could, and often

did, go on for hours about something in a quite serious

tone, anyone who spent much time with Bill knew that

he had a wicked sense of humor, and conversations with

him could swing between debates about the International

Code of Zoological Nomenclature one minute to a

hectic exchange of friendly insults the next. That he was

known as “Uncle Bill” to many speaks volumes about

how comfortable we all felt with Bill. My relationship

with him was somewhat different. Years ago I told Bill

that I thought of him as the older brother I never had and

indeed, the last words Bill spoke to me were “Be well,

little brother.” We are all both better herpetololgists and

better people for having known Bill.

Stephen Spawls

Although Bill and I had corresponded since the early

1980’s, we didn’t meet until 1987, when Bill drove up

to Botswana and stayed a few days with us at Moeding

College, Otse. It was an exciting visit. Bill came in a

white windowless Volkswagen Kombi, which was the

same type of vehicle that had been used by the South

Africa Defence Force on their 1985 raid into Botswana.

Consequently, the Botswana security forces had tracked

the vehicle, and as Bill drove out of our college he was

stopped by the soldiers, who went through the vehicle.

Finding nothing, the military concluded that Bill had

cached his weapons at my house which was then

searched! After this inauspicious start, my wife and I

subsequently stayed with Bill in Port Elizabeth. We went

on an amazing safari, to the Addo Elephant National

Park, to Graaff-Reinet, and thence into the Karroo, where

we stayed at the Karroo National Park headquarters with

Bill’s friend, the warden Harold Braack. We returned via

the Swartberg and Oudtshoorn.

Being in the field with Bill in some of his favourite

country was an amazing experience; he knew the land, the

customs, and the animals, and gave freely of his expertise.

We found many spectacular species that were totally

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new to me, including Hewitt’s Ghost Frog, Heleophryne

hewitti, the Giant Ground Gecko, Chondrodactylus

angulifer, and the Blue-spotted Girdled Lizard, Ninurta

coeruleopunctatus. But we weren’t lucky all the time. At

one point Bill and I drove for several hours at night on the

tarred road near Beaufort West, hoping to find a Horned

Adder, Bitis caudalis, but we saw virtually nothing. As

we returned to the park, near midnight, there on the road

was a snake, and we leapt out with great excitement but

it turned out to be only a Herald Snake, Crotaphopeltis

hotamboeia. One morning Bill and I were pursuing a sand

lizard, Pedioplanis, which was sheltering 1n a clump of

bush. As it appeared near my feet I dove at it but missed,

and it fled to another clump. Bill clapped his hands on his

head. ‘It’s obvious you’ve always collected by yourself’,

he said exasperatedly, ‘you should have just shuffled it

towards me, not leapt at it without telling me.’

On that trip, we also learnt of each other’s shared

enthusiasm for bird-watching. As we drove across

country, Bill directed me to a side road. ‘I’ve got a

surprise for you,’ he told me, as we took the diversion.

We went a few miles and then Bill told me to pull up and

get my binoculars; and there, in a grassy area below the

road, were a pair of Blue Cranes, the first I had ever seen.

In 1991, I went to work in Ethiopia, and Bill wrote

to me in 1992, suggesting we might work together on

a book on Africa’s dangerous snakes. Blandford Press

showed interest in the project, and in 1993 Bill came

and spent a few weeks with us in Ethiopia, doing field

work and working on the book. We made several field

trips, one was to the highlands east of the Rift Valley,

to a town called Dodolla where a specimen of Bitis

parviocula, the spectacular Ethiopian Mountain Viper,

had been collected, still the only specimen known from

east of the rift valley. As we ascended the rift valley

wall, up through dense broad-leafed forest, we became

increasingly excited; this looked like Bitis parviocula

country. Then as we approached Dodolla, we emerged on

the plateau, and found ourselves on a vast open grassland,

as bald as a billiard table. Bill sighed and looked at me.

‘Listen, matey’ he said (Bill and I were both born in

North London, he at Finsbury Park, me in Muswell Hill,

and sometimes in the field we were just two Londoners

together), ‘we’re looking for a forest viper, and as far

as habitat goes, we’ve just gone from the sublime to the

ridiculous.’ But that day we did find some spectacular

frogs, including Paracassina kounhiensis, Mocquard’s

Mountain Kassina.

The following week, down in Awash National Park,

we had some remarkable luck; in one afternoon and

evening we got a North-east African Carpet Viper (Echis

pyramidum) under a rock right outside our room, on

the road in the dark we found a Kenya Sand Boa and

two species of egg-eater, and as we drove back to the

lodge, we caught a huge Atractaspis fallax on the road,

an adventure that Bill described as being ‘like trying to

subdue a spiked manhole cover.’ At Lake Langano, we

Amphib. Reptile Conserv.

xix

caught a small Egyptian Cobra (Naja haje) on the road.

Back in Addis Ababa, I was teaching one morning and

Bill worked in the garden, creating a small set on top

of a rather nice garden table to photograph the cobra.

Unfortunately, he incorporated several biggish rocks

in the set and in moving these around, he managed to

thoroughly gouge the polished surface of the table. My

wife went ballistic, but Bill turned on the charm and

managed to persuade her that it was all part of the great

scientific endeavour, and he took us out for a meal as

well. The following day I found Bill crawling around in

the canna lilies when I got home; one of the frogs he

was photographing had sprung into the flowerbed and

escaped.

We didn’t always get on well. Bill had a very relaxed

attitude towards deadlines, and often preferred to go into

the field rather than knuckle down. He once told me how

his publishers (Struik) ‘had flown him to Cape Town’

to finish his field guide, and a fellow herpetologist,

who overheard this, said ‘What you mean, Bill, is that

Struik made you fly there, sat you down in their offices

and said you weren’t leaving until you got it finished.’

Bill laughed and admitted it; and in one of his books he

thanks his editor for ‘making ridiculous deadlines seem

acceptable.’ Our work on the Dangerous Snakes of Africa

book was complicated. Bill was in South Africa, I was in

Ethiopia, and there was no e-mail in those days. We used

to send stuff to each other by courier. As the deadline

for the delivery of the manuscript approached, Bill had a

lot of the snakebite stuff still to do and wasn’t getting it

done. With two weeks to go and the publishers muttering

angrily about penalty clauses (the production was

catalogued, and tied into a publicity/release schedule), I

rang Port Elizabeth to be told that ‘Dr. Branch had left on

an extended safari to Zambia, and would not be back for a

few weeks.’ In a panic, I managed to get hold of Dr. Colin

Tilbury, who stepped into the breach and wrote virtually

all the snakebite stuff in short order. The manuscript went

in on time, but it led to a furious row between Bill and I

over the order of our names on the cover. But eventually

we got over it, and in fact in 2017, we agreed to do a

revision of the Dangerous Snakes book.

The last time I met Bill was in 2014, in Bagamoyo,

Tanzania, where we were part of a team assessing

Tanzania’s reptile biodiversity. We went out one evening

and I climbed a tree to catch a sleeping Spotted Bush

Snake. Bill watched thoughtfully. ‘I’m past climbing

trees’, he told me. ‘In fact, ’'m past climbing over

anything. Last time I was in Namibia with Aaron Bauer,

a lizard ran under quite a low fence and neither Aaron

nor I could get over it.’ He laughed, ‘It’s my fondness

for prawn curry.’ On that conference, Bill talked with

great enthusiasm of a projected book. ‘I really want to do

a big book’ he told me, ‘covering the natural history of

Africa’s snakes, along the lines of Harry Greene’s book.’

He showed me some ideas and pictures on his computer;

his ideas were mind-stretching and holistic; he saw

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Tributes to William Roy Branch (1946-2018)

Fig. 26. Bill Branch photographing some lilies at Lake

Langanao, Ethiopia (Photo: Steven Spaw!ls).

the snakes in the landscape as part of the interlocking

whole ecosystem, and his accompanying pictures were,

as always with Bill’s photographs, spectacular. Nobody

else has photographed the African herpetofauna like Bill.

We started work on the revision of the Dangerous

Snakes book in early 2017. Bill sent me some draft

material, a list of important references (he had an

encyclopaedic knowledge of the literature on African

reptiles), and a stunning portfolio of pictures. But in

late 2017, Bill cautiously wrote to tell me he was having

mobility issues, and wasn’t sure how this might affect

our project. And in early 2018, to my shock, I heard

from Bill that he had been diagnosed with motor neurone

disease. But he remained full of optimism; and said that

he fully intended to do his bit; cheerfully pointing out

that Stephen Hawking had lasted many years with the

same affliction. But it was not to be. The disease quickly

took hold. Tragically, Bill died on the 14th October

2018. His untimely death is a major loss to African

herpetology. And I hope that our book, which should be

published in mid-2020, will be a suitable monument to

Bill. Few herpetologists have reached both the public and

their fellow scientists with such verve and accuracy as

Bill did.

Se Sc] Miisieees eos eS aa

Fig. 28. Bill Branch and others at Bagamoyo, Tanzania, in 2014.

Amphib. Reptile Conserv.

Fig. 27. Bill Branch admiring an old tank near Dodolla,

Ethiopia (Photo: Steven Spawls).

Andrew Turner

CapeNature, Western Cape, South Africa

I first met Bill Branch at an HAA meeting, I think the

Stellenbosch meeting of 1998 or thereabouts. He was the

top South African herpetologist in my mind because of

his comprehensive treatment of the reptiles of the region

(his lesser interest in amphibians did not bother me, as

his emphasis on the snakes more than made up for this).

He was always interested in other people’s experiences,

especially regarding observations on distributional

occurrence, and encouraged documenting this valuable

form of data. Knowing that someone like Bill, who

came from a rather different background, could switch

to making a career of herpetology—and a rather exciting

and enjoyable career at that—was a great inspiration for

me to continue my professional herpetological interest.

His photography was also inspirational and keeps me

(and many others) clicking away.

Bill was a great raconteur, and his stories of herping

gone wrong were particularly amusing. One story

in particular, although I don’t remember that exact

mechanics of it, involved Bill trying to catch a Cordylus

under a rock that was being lifted by a colleague. (I shall

not mention his name but he did have an extensive snake

collection at one point). Bill shoots his right hand under

the rock to catch the Cordylus but notices at the same

time there is a second Cordylus under the rock so shoots

in his left hand too, catching both of them. But then a

third Cordylus runs out from under the rock and said

colleague catches that one by dropping the rock on both

Bills hands!

Bill really set the scene for getting the full picture of

South African herpetological diversity, and did a good

job of placing this in an African and global context. He

travelled widely and shared his great photographs, and

was always wondering how the various species fitted

together. His fondness for the small adders was totally

understandable, and he did a good job of discerning their

subtle (and probably recent) divergence and highlighting

the need for conservation of several of these species.

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eo . = = Sp a h °

Fig. 29. Bill doing what he did best: geeing everyone up to

maximise income from the HAA auction! (Photo: Andrew

Turner).

Ernst H.W. Baard

CapeNature, Western Cape, South Africa

My career as a herpetologist with CapeNature (then Cape

Department of Nature and Environmental Conservation)

started in January 1983. My first task was to sort out

and process a few thousand specimens of frogs, lizards,

snakes, and tortoises collected and collated by my

predecessor, John Comrie Greig (Greig and Burdett,

1976. Patterns in the Distribution of Southern African

Terrestrial Tortoises) and, at the time, colleagues,

Richard Boycott and Atherton de Villiers.

After writing (yes, there were no emails those years)

to the curator of the Port Elizabeth Museum, Dr. Bill

Branch, and the curator of herpetology at the South

African Museum in Cape Town, Dr. Geoff McLachlan,

about depositing the specimens (roughly divided into the

Western and Eastern Cape), we got positive responses

from both curators. Atherton and myself proceeded, and

we completed the task of sending, among others, the

whole Greig and Burdet wet and dry tortoise collection,

and several hundred “eastern” Cape lizards, snakes,

and frogs to Bill at the Port Elizabeth Museum. Bill’s

epic paper on the lizards of the Cape Province (Branch,

William. 1981. An Annotated Checklist of the Lizards

of the Cape Province) made a huge impact on my

knowledge of the lizards of the Cape, and together with

FitzSimons 1943 (Lizards of South Africa) guided us

through the process.

An incident that stood out during this time, was

the discovery in the Jonkershoek collection of an un-

identified many-spotted lizard in a small bottle, collected

in 1973. It was beautifully preserved and, fortunately,

with a geographical location in the Groot Winterhoek

Mounatins down to seconds South and East (this was

before GPS). It took me a few days using FitzSimons

1943 to identify the lizard as “Lacerta” australis, and we

were very excited about this discovery. One unsuccessful

collection trip to the locality (by Boycott, De Villiers, and

Baard) was undertaken in February 1983, followed by

Atherton and I managing to collect two more specimens

Amphib. Reptile Conserv.

at the same locality in April 1983. Imagine our joy, since

these were as far as we could establish, only the 6th and

7th specimens known to herpetology of this “elusive”

species. This information was shared with Bill and he

encouraged us to publish a note which we promptly

did, asking him to co-author (De Villiers, Baard and

Branch. 1983. ‘Lacerta’ australis: additional material).

Bill was always supportive of any further investigations

and readily responded to queries on the herpetological

collection.

In later years, Bill got back to us and was very excited

about some of the tortoise specimens we sent him,

including some of the largest individuals of some species

he had encountered. He then published a short note in the

Journal at the time, honouring Atherton and I with co-

authorship (Branch, Baard and De Villiers. 1990. Some

exceptionally large Southern African chelonians).

I only met Bill for the first time at my first HAA

Conference in Stellenbosch a year or two later, and was

really honoured to make his acquaintance. His paper on

angulate tortoise ecology in the Eastern Cape (Branch

1984. Preliminary observations on the ecology of the

Angulate Tortoise) had a huge impact on my career,

since this paper shaped my thoughts and guided my

research and attempts at understanding the ecology of the

geometric tortoise of the Western Cape; having completed

my research in 1990. For a young herpetologist like me at

the time, it was almost natural to think: What would Bill

do in this case? or How would Bill approach this topic?

Bill’s astonishing knowledge of lizards, tortoises

and snakes, snake venom, snakebite, etc. was really

something to behold, and few herpetologists could keep

up with him. I fondly remember Bill at conferences

communicating with all and sharing his knowledge. The

best story I remember him telling, was about the evening

in the veld around the fire. After Marius Burger latched

a Pseudocordylus crag lizard to his (Marius’) earlobe,

Bill kept on touching the lizard which wouldn’t let go

of Marius’ ear; with the lizard biting down harder and

harder, Bill spent an hour or so enjoying Marius’ agony

and futile attempts to get the lizard to let go of his ear!

William R. (Bill) Branch was a legend of his

generation and time. Not only was he a brilliant scientist,

excellent herpetologist, and I believe, a great bird

enthusiast, but also somebody one could look up to. His

contribution to South African and global herpetology

will go down in history as exceptional, ground-breaking,

and outstanding, and will stand the test of time like with

other greats; FitzSimons, Broadley, etc. His contribution

to the written and peer-reviewed herpetological science

and popular literature is unsurpassed, and it is my honour

to have known him.

Amber Jackson

Cape Town, South Africa

Uncle Bill’s Bible was a well-used field guide by the

time I met the man himself. I had even sent him a few

September 2019 | Volume 13 | Number 2 | e186

Tributes to William Roy Branch (1946-2018)

Fig. 30. DNA sampling of Spek’s Hinged Tortoise, Kinixys

spekii (Photo: Amber Jackson).

specimens of Leptotyphlops years before when I was

a student. I finally met Bill as an awe struck herpie

requesting he sign my copy of his field guide before a

very serious meeting at my new place of work. He wrote:

“Wow, Uncle Bills bible!!! 167 species out of date, but

what else is there.” Out of nowhere he then spouted a

lecture about the Galapagos and island biogeography,

and held up the meeting for 20 minutes in the process.

All eyes on me, I left with my signed copy and some

knowledge I never requested but was all the better for

knowing. Little did I know, our first meeting was an

accurate precursor for the years that followed. Thanks

to numerous development EIA’s Bill and I travelled to

Lesotho, Augrabies, and Mozambique (multiple times),

with me always as his self-proclaimed assistant.

One of my favourite memories with Bill is lying in

the dark, on a rocky shelf at the top of the Augrabies

paleo falls, staring at the stars and waiting for the

geckos to come back out after our disturbance. The stars

were incredible! I later caught him a Pachydactulus

atorquatus, without breaking the skin, and received an

exclamation of ‘I could kiss you.’ He didn’t, and ran off

with his prize. At the time, I was naively more excited

that 1t meant we could go to bed before our dawn wakeup

call in four hours. Bill, 40 years my senior, put me to

shame with his energy levels.

We got along at first because I was eager to learn,

and he was eager to teach. Then one day, I called him

a bastard for one of the anti-feminist comments he used

to purposefully provoke me with, to which he laughed,

Amphib. Reptile Conserv.

Fig. 31. Bill reading science in 45 °C heat (Photo: Amber

Jackson).

sighed, and said “finally!” All pretenses over with, we

were then friends.

Most of my time with Bill was spent learning, so

much so that my brain stopped being able to absorb

any more information and saturated by the end of the

long day. The knowledge he possessed was impressive,

diverse, and felt insurmountable. He taught me plenty

about herpetology and science in general. His enthusiasm

was contagious and witnessing his studiousness in the

field was impressive, with his daily diary and specimen

processing. But one thing that stands out is the things he

probably never intended to teach me, for example: How

being reserved doesn’t have to impact negatively on your

life, how euthanizing something as cute as a bush baby

can make a huge contribution to science and someone’s

career, or how LED lights can make fresh produce more

appealing. How just because your life starts in one place

doesn’t mean you have to stay there. How you can be a

jack of all trades and a master of one. How you can make

mistakes. That apologies are important. How careers can

be diverse and often unfold. That one of the biggest joys

is to love, freely and openly.

To the man that “was by turns (and somehow all at

once) relaxed, intense, sincere, self-mocking, modest,

confident, serious, and funny.” Kim Stanley Robinson

To the man that could make an economist understand

biodiversity by using economic terms.

To the man that could answer the question: “Is a

penguin a fish or a bird?” politely and honestly.

To the man that could provoke an entire lecture with a

simple question: ‘What’s the odd one out?’

To the man that believed in love, in science and in the

unknown.

To the man that studied cancer but fell in love with a

cobra while fishing.

To kind, funny, and sometimes forgetful ‘Bum in the

Butter’ Bill.

I think about you often, your teachings, your adventures,

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Fig. 32. Bill with a shoftshell terrapin he caught (Photo: Craig

Weatherby).

¥ fe a! 7h? 7

ov ; / ak d a

our last day together, and all the days before then. I couldn’t

have asked for a better mentor, teacher, and dearest friend.

I will treasure you always. Thank you for believing in me.

Margaretha Hofmeyr

University of the Western Cape, South Africa

I first met Bill Branch in October 2000, when my

colleague Alan Channing invited him along on a field trip

to Namibia for the UWC Zoology Honours students. I had

most students with me in my husband’s Kombi Synchro,

while Bill was a passenger in Alan’s Land Rover. We

stopped at Springbok’s Caravan Park for the first night,

where I booked accommodation in chalets. Because Bill

joined the party at the last minute, I knew there would not

be a bed for him and had some concerns about sleeping

arrangements. For one or other reason, perhaps because

Bill was idolised by all herpetologists, I expected him to

be rather arrogant, but he quickly won me over when he

made a bed for himself in the trailer Alan took along. The

trailer was quite short, but so was Bill; fortunately the

trailer was rather wide, because so was Bill. The sight of

him surfacing the next morning from his trailer bed will

always stay with me. Yes, he might have been arrogant

at times, but he was always a great sport, and teased the

students to distraction on this trip.

I always feel dishonest calling myself a herpetologist,

because my field of expertise is restricted to tortoises

and terrapins. Yet, on this trip, as herpetologists do, we

went on night drives to look for herps (never tortoises)

on the roads. One of the nights while staying at Klein

Aus, everybody squeezed into my Kombi to search for

exciting things on the roads. While driving through a

narrow stretch of road between two fences, a springbok

ram materialised in the road before my husband’s car. I

switched the main lights off within seconds, but it was

still too late. The springbok ran straight into the Kombi,

broke his neck, and put quite a dent into the front of the

car. All the girls were crying but we had to deal with the

situation. Alan and Bill dragged the springbok out of

the road and then we had the unfortunate task of driving

to the owner’s house to report the incident. His only

Amphib. Reptile Conserv.

reaction was that it was the only springbok he had on

the farm. This was an unpleasant experience for all of us

but also created a bond, because Bill mentioned it many

times to me in ensuing years.

At conferences, I would get annoyed with fellow

herpetologists for teasing Bill about his lisp, yet, he

always laughed at their jokes. To me, the ability to laugh

at yourself reflects true character, and Bill had that. I have

many fond memories of Bill and always regarded him as

the ultimate herpetologist and naturalist in South Africa.

His expertise stretched so much wider than reptiles and

amphibians. He may not have been an expert on every

animal or herp group, but his knowledge was astounding.

He was also willing to share his expertise and helped

many young scientists to find their way. I may not be

described as a young scientist, but when I switched from

large mammals to tortoises, Bill knew much more than I

did, and he was willing to share. Over the years we co-

authored several papers and it was always a pleasure to

work with him in a professional capacity. South Africa,

Africa and the World are now deprived of one of their

top intellectuals, and an exceptional person—we salute

you Bill.

Jens Reissig

Ultimate Creatures, Gauteng, South Africa

The first time I met Bill was during a high school field trip

to northern KwaZulu-Natal around the year 2000. At that

stage, I was rather shy and having had a very keen interest in

reptiles since my early childhood I of course knew exactly

who he was, however never made any contact with him.

Many years had passed until I crossed paths with him again

at the Herpetological Association of Africa’s Conference at

the National Zoological Gardens in Pretoria, South Africa.

From this point on, we stayed in contact and he was always

willing to help wherever he could. Unfortunately, he was

extremely busy while I was compiling my book on the

Girdled Lizards and their Relatives in 2013 and 2014, so

that he was not able to assist me with it in any way. He

did however end up reviewing the book for me. The book

review ended up being published in Herpetological Review,

2015, 46(2): 1-7.

After having received the tragic news of Bill’s

diagnosis, I decided to go and visit him at his home in

Port Elizabeth on the 20th of April 2018. Even though

one could see that he was battling his illness, he still tried

to be upbeat about life and could not stop talking about

reptiles and a (to me) hidden passion of his, Orchids. We

sat for hours on his patio talking about various reptile

projects, his Orchid collection, birding, and some of his

many field trips into Africa. After spending quite some

time with us, one could see that he was tiring and we

decided to say our goodbyes and I left. It was the best

day I had ever spent with him. My favourite email I have

ever received from him was received on the 12th of June

2015 and stated: “Dear Jens. Here are the proofs of my

review of your excellent book which should appear in

September 2019 | Volume 13 | Number 2 | e186

Tributes to William Roy Branch (1946-2018)

.—

Fig. 33. 2017 Herpetological Association of Africa’s Conference

in Bonamanzi, South Africa. From left to right: Werner

Conradie, Tyrone Ping, Prof. William Branch, Luke Verburet,

Dr. Michael Bates, Johan Marais, Prof. Graham Alexander,

Prof. Aaron Bauer, Jens Reissig, Coleen Tiedemann, Dr. Colin

Tilbury, and Dr. Victor Loehr (Photo: Andre Coetzer).

Herp Review this month. Hope you're happy with it and

all goes well. Best wishes. Bill”

Professor W.R. Branch, your passing has left a

massive hole in so many people’s lives and in African

Herpetology as a whole. Your knowledge and sense of

humour will be greatly missed by anyone who ever had

the privilege to know you and who’s life you may have

affected in some way or another. Africa has lost two

great herpetologists way too soon and in relatively quick

succession. Till we meet again!

Harold Braack

South Africa

Bill Branch first came into my life, I think, in 1974,

at an HAA conference held at Skukuza in the Kruger

National Park (KNP). It might have been the first such

get-together. At that time, I was doing the herpetology

survey of the KNP, so it was indeed fortuitous to meet

Bill Branch, as well as Don Broadley and Carl Gans.

In 1976, I was transferred to the Bontebok National

Park and so I started species surveys and checklists. I

wanted to know what it was that I was supposed to be

looking for, and so Bill and I started working together

through various National Parks and adjacent areas. But

it was not only herps. We looked at succulent plants and

birds as well.

What Bill gave me was the confidence to do the

surveys. In him, I had a partner with whom I could share

my passion for conservation and protection of all the

inhabitants of those areas. He was totally enthusiastic

and this rubbed off on us all. Above all, he was a good

friend.

I have many fond memories of Bill, but share only a

few here.

The pepper ticks at Addo National Park were a vast

irritation to Bill. We picked them up every time we

ventured on a collecting trip. He hated them. I wiped

them off with paraffin. But Bill had a different solution.

Amphib. Reptile Conserv.

“Oh no,” he said. “I go home. I get undressed completely

then lie down in the nude on the kitchen counter. My

family has to pick them off. “

Bill Duellman and Bill Branch stayed with us in the

Karoo for several days. To separate the two, we called

Duellman “Bill” and Branch “Billikins.” Bill B. was not

totally enamoured with the solution.

Bill and I did a night road survey in the Richtersveld.

After several hours we arrived at Paradyskloof. We lay

down flat on our backs for a while counting passing

satellites, then later scratching among the rocks where

we saw the largest Hadogenes that either of us had seen.

Then Bill went to the little pool to find a Strongylopus

springbokensis. Ka-splash, splash. Bill fell in the pond.

He sat huddled in the bakkie on the way home.

Spending a long, long time trying to catch a lizard in

Richetrsveld, Bill suddenly ran back to the bakkie. Out

he came with a revolver loaded with dust shot. “Ka-

boom!” he shot the thing—we had our specimen.

He had excellent repartee and a quick lucid mind.

How many of us remember his response during a frog

meeting at Stellenbosch? The chair said we should be

democratic in the course of the meeting. Bill’s immediate

response “Thank You, Mr. Mugabe.”

Bill also enjoyed fishing, especially for carp. We spent

some time along the banks of the Orange and Breede

Rivers doing just that. Didn’t catch much, but those were

relaxing times.

I best remember Bill as a man who was a dear friend.

To all of us, he revealed the treasure chest of our vast

herpetological wealth—and, more, he opened it up for us

to see and explore. He followed his passion with a radiant

glee which he passed on to us.

Bill, my friend, I salute you for being a friend, a guide,

and an explorer who found and revealed.

Atherton de Villiers

CapeNature, Western Cape, South Africa

I have good memories of Bill that date back to when his

career 1n herpetology started at Port Elizabeth Museum,

and have always admired his enthusiasm and vast

knowledge of reptiles and amphibians. It is well known

that one of his greatest achievements was his Field Guide

to the Snakes and other Reptiles of Southern Africa. This

landmark publication opened up the world of reptiles to

countless numbers of people, and it was a pleasure to

contribute information and images for one of the most

important herpetological publications in southern Africa.

I share with you all the huge loss of Bill to herpetology,

biodiversity conservation, and life in general.

Marius Burger

North-West University, South Africa

Try as I may, I just can’t seem to pinpoint the precise

memory of actually meeting Bill for the first time. ?'m

quite shocked by this realisation. I presume that it was

sometime during 1987 when I was a young (20 y/o)

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Fig. 34. Ozzy Osbourne meets Elvis Presley. My all-time

favourite photo of Bill and I (Photo: John Measey).

nature conservation student in Grahamstown, and I

vaguely remember something about visiting him at

his office at Port Elizabeth Museum (PEM). The only

definite lead that I have to go on is a specimen of Karoo

(ex-Namaqua) Plated Lizard (Gerrhosaurus typicus)

that I collected in November 1987 on the Karoo Nature

Reserve in Graaff-Reinet. At the time, this record

represented an eastern distribution range extension

of 211 km. Whoopee! I would have hurriedly taken

the specimen to Bill at PEM, with my tail wagging in

excitement. Yes, what a joy it always was to make some

sort of new herpetological discovery that even The Bill

Branch would find somewhat noteworthy. And so it came

to pass that my first ‘scientific publication’ was a short

note in the Journal of the Herpetological Association

of Africa (Burger 1988). The truth be known, Bill

actually wrote the damn thing. But this was my official

introduction to the Herpetological Association of Africa,

and it marks the approximate start of a very lekker 30-

year friendship with Bill.

An article published in Zootaxa on 24 October 2018

demonstrated that the African Slender-snouted Crocodile

(Mecistops cataphractus) is in fact comprised of two

superficially cryptic species, and thus M. leptorhynchus

from Central Africa was resurrected as a valid species

(Shirley et al. 2018). The first thought that crossed my

mind when I read this paper was “Fok, Bill didnt get

to see this!”, because Bill had died ten days earlier. Bill

would have loved the news that Mecistops is monotypic

no more, and perhaps (probably) he even knew that this

was in the pipeline. Fast-forward nine months to July

2019 (i.e., right now as I’m writing this), and I’m still

experiencing Where-TF-is-Bill moments on an almost

daily basis. Bill was my Google Herps. Whenever

I needed photos of far-flung African reptiles to be

identified, the oh-so-convenient Google Herps would

Amphib. Reptile Conserv.

Fig. 35. Bill emerging from swamp in Gabon (2002) after

checking funnel traps that he had set with the hope of catching

an African Parachanna (Photo: Marius Burger).

usually be my first check. When the complexities of

taxonomy would bewilder my brain, Bill had the knack

of explaining it in a way that I could sort of comprehend.

And so for me, it is not only very sad, but also utterly

inconvenient and totally kak that Bill died.

It always intrigued me that a Pom could arrive in

Africa with some sort of medical doctorate degree,

something to do with foetal rabbit liver metabolism and

primary liver cancer, only to end up perusing a career of

chasing reptiles and amphibians. Like, how the hell did

that happen?! Anyway, it’s a good thing that it turned out

the way it did. Well, so says I, because I have derived

much joy and intellectual enrichment from the times

hanging out with Bill. To South Africans, Bill was loved

and respected as our local herp guru. He was of course

also internationally renowned for his herpetological

contributions, and the momentum that he built up over

the decades will have him publishing papers for a long

while after he clocked out.

Fig. 36. Bill receiving medical attention during a biodiversity

survey of Loango National Park (2002). If I remember correctly,

it had something to do with removing ticks from a place where

the sun don’t shine (Photo: Carlton Ward Jr.).

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Tributes to William Roy Branch (1946-2018)

This may be somewhat of a narcissistic trait of mine,

but I like it that Bill liked me. I would often purposefully

say and do socially objectionable stuff in Bill’s presence,

for the reward of his approval and appreciation of

my crudeness. Now that I think about it, this kind of

behaviour was probably akin to a son showing off in

front of his father for attention and approval. In a book

that collated a collection of interviews on how to become

a herpetologist (Li Vigni 2013), Bill wrote the following:

“T also never miss a chance to be with Marius Burger,

just to re-emphasize how sane I am.” Whenever Bill

acknowledged me in a publication, I would smile, and

feel all warm and gushy inside for receiving his praises.

In the acknowledgments section of Tortoises, Terrapins

& Turtles of Africa (Branch 2008), Bill wrote: “A special

thanks to Marius Burger, whose tortoise photography is

Just too good.” Well, just imagine the grin on my face for

that bit of flattery. He then went on to say: “...ifhe could

only look after his camera lenses as well as his hair...”

in The Dangerous Snakes of Africa (Spawls and Branch

1995), and Bill included a thanks to a certain Marias (sic)

Birger (sic) for companionship and advice. If that was

indeed me that he was referring to, then I say ditto to that.

Whilst on a fieldtrip with Olivier Pauwels and Bill

Branch in Gabon, the three of us shared a shipping

container that was modified into a bedroom of sorts. I

retired to bed late one evening, with Olivier and Bill

giggling away like preteen girls. The reason for their

hysterics was that they had planted a condom half-filled

with Condensed Milk in my bed. How silly 1s that!

Anyway, I never noticed said condom in my bed and

managed to fall asleep in spite of the spurts and snorts

of laughter. The next day whilst checking our trap arrays

they told me of their really funny prank, all the while

grinning from ear to ear as they awaited my reaction.

Instead of shock and dismay, I replied with a calm

reminder that a cleaning team was making our beds each

morning and just imagine what their take would be on

discovering this soggy item in one of our beds. I watched

as their expressions gradually turned from smile to mild

alarm, as the two of them slowly processed and realised

the gravity of this scenario. Now it was my turn to laugh.

I’m not a spiritual kind of guy, and thus I won’t be

saying things like RIP. old friend or check you on

the other side. But ja, Bill was for sure a significant

component of my life. Iam very chuffed to have had him

as a friend.

Mike Raath

Director, Port Elizabeth Museum Complex (now

Bayworld), 1987-1995

I first met Bill Branch in the early 1980s, when I was

at Wits University as head of the Bernard Price Institute

for Palaeontological Research. I had been invited by

Prof. Brian Allanson of Rhodes University’s Zoology

Department to present a short course on the evolution

of the Class Reptilia at my much loved Alma Mater.

Amphib. Reptile Conserv.

At that early point in my career, I only knew of Bill

by reputation, and had never met him personally. I felt

flattered that he had taken the time and trouble to travel

from Port Elizabeth to Grahamstown to listen to my

ramblings. I little realised then that he and I would meet

up again in a different context several years later, when I

was fortunate enough to be appointed Director of the Port

Elizabeth Museum Complex, as it was then called before

it got its trendy current name of “Bayworld.’

Bill was a much-respected member of the research

staff of the Complex, having charge of one of the most

comprehensive and important herpetological collections

in the country, building on the solid legacy of its original

founder, the legendary F.W. FitzSimons, almost a century

before. He was one of the stars of our research team,

regularly producing work that was published in some

of the world’s top peer-reviewed scientific journals. But

in addition, he was a prolific writer of popular articles

and books aimed at the general reader that spread his

expert knowledge to a much wider general readership.

I remember one envious member of our research staff

calling him “the Naas Botha of our research team” in

terms of earning brownie-points for research output (only

those who know something about South African rugby in

the 1980s will understand that comment! ).

One of the things that defined Bill was his off-the-

wall unconventionality. I remember how audiences at

his various public presentations would shudder in shock,

horror, and jaw-dropping disbelief when he demonstrated

his go-to technique for distinguishing between identical

sibling species of toads—by licking them! And, by Jove,

it worked!

I respected Bill as a person and as a scientist. But to be

candid, I have to say that he and I did not get on that well

personally. He suffered neither fools nor administrators

gladly, so as his director I guess I failed on both counts!

But as a scientist committed to his discipline there was

no faulting him. He was single-mindedly devoted to his

collection and his research, often to be found in his lab

or office over weekends or public holidays when most

others on the staff were taking what they rightly regarded

as a well-earned rest.

One particular Saturday morning remains starkly and

darkly etched in my memory, when Bill received an urgent

call in his lab mid-morning from the Snake Park. One

of the Snake Handlers, Mr. Nimrod Mkalipi, had been

bitten by a Puff Adder at the end of the daily live snake

demonstration, and he was in dire distress. Bill dropped

everything and rushed to Nimrod’s side, administering

antivenom and applying appropriate emergency first

aid. Tragically, though, it was to no avail and Nimrod

succumbed on the scene despite Bill’s urgent, expert, and

devoted efforts. Medical opinion afterwards held that it

was anaphylactic shock that took Nimrod’s life, and that

nothing other than immediate on-site specialised medical

intervention would have had any chance of preventing

it. That event shocked us all. It is a dark memory that I

September 2019 | Volume 13 | Number 2 | e186

Conradie et al.

I si ee ee ee

Fig. 37. Bill walking off into the early morning light to

photograph some Welwitschias in south-western Angola in

2009 (Photo: Werner Conradie).

carry with me to this day, but I applaud Bill Branch for

his swift reaction and his valiant and urgent attempts to

save the life of a fellow staff member. We all had much

to learn from that tragedy.

Werner Conradie

Port Elizabeth Museum, South Africa

The first time I became aware of Bill was when I

attended my very first HAA conference at Bayworld in

2004, which he organised. I remember the occasion very

clearly, as the only options available at the icebreaker

were beer, and Coca-Cola. I was too shy to speak to him

then—he was the famous Bill Branch and I was, after

all, just a lowly student. I met Bill again at the 2006

HAA held at Potchefstoom, and this time I was on the

organising committee. At this event I recall fondly Bill’s

talk on ‘guts and gonads,’ which of course went well

over its allotted time. However, it wasn’t until I finished

university completely that we would have what would

turn out to be a bit of a prophetic chance encounter.

While on a December break, just before I would start a

new job as a Physical Science high school teacher, me

and my now wife walked into him while strolling around

the museum. I introduced myself to him, finally having

a moment in his direct eyesight after all this time, and

with a curt nod paired with a brief “nice to meet you,” he

disappeared through the door to his lab and office. Never

would I have guessed that less than six months after this,

I will be sitting in front of him for an interview for the

job of Assistant Herpetologist. I must have impressed

Bill one way or the other (couldn’t have been pure

desperation), as two days later I received a call that I got

the job. I finished my contract at the school as fast as I

could, and with great excitement walked straight into the

museum the very same day. Bill looked at me in utter

shock and sent me home, saying I should come back in

the New Year... I guess he wasn’t prepared for my quick

start!

For the first year at the museum I had to learn all

the ropes. Now this is a very steep uphill battle for

Amphib. Reptile Conserv.

Fig. 38. Bill holding a Meller's Chameleon (7rioceros melleri),

looking radiant despite a very tiring hike during the summit of

Mt. Mabu in 2009 (Photo: Werner Conradie).

an Afrikaans-speaking seun that knew his frogs, but

no reptiles. Up to that point in time, the only reptile

ever caught by me was a harmless Common Brown

Water Snake. The first challenge I faced was getting

into a conversation with Bill. Because of his British

heritage, he mumbled a lot and this made me struggle to

understand his pronunciation of scientific names. After

many “conversations,” I often went back to my office

and paged through his field guide to prepare for the next

engagement. Bill, however, never forced his reptilian

inclinations on me. Once, he walked into my office and

promptly asked me what I wanted to specialise in, to

which my response was tadpoles. He dryly remarked that

the only thing they are good for is fish bait. As it turns

out, I never did work on tadpoles that much...

I went into the field with Bill for the first time as part

of a multi-collaborative expedition to Angola in 2009.

Bill didn’t have to bring me along, he could have kept

all the new places and specimens to himself, but I will

ever be grateful as it was on that trip that I fully came

to understand and realise what my responsibilities as a

museum herpetologist include: New discoveries! I joined

Bill on two more consecutive trips to Mount Mabu,

Mozambique in 2009 and Lagoa Carumbo, Angola in

2011. It was at this stage that Bill assisted me with my

first-ever species description, and just as I was starting to

bask in the glow of his knowledge, he clearly thought he

had trained me enough and turned off the light. It seemed

I was on my own: Bill expected me to swim. It was up to

September 2019 | Volume 13 | Number 2 | e186

Tributes to William Roy Branch (1946-2018)

me to show him that I could. We wouldn’t go on another

field trip together again until 2015, again to Angola. By

this time, Bill had retired, and I was keeping the fort on my

own. During the trip, around the campfire one evening,

Bill told me that he can now rest in peace, knowing the

Port Elizabeth Museum herpetology collection is in good

hands. Thank you, Bill.

I worked with Bill for more than ten years, but I only

really started to get to truly know him and his family

when he was unfortunately diagnosed with motor neuron

disease. It was a devastating experience to see your

mentor and friend fade away in front of your eyes. Bill

was determined to follow his passion to the very end, and

his determination was amazing to behold. Bill has taught

me life lessons that I will cherish forever. He was truly a

one-of-a-kind man. He is and will be missed.

Martin J. Whiting

Department of Biological Sciences,

University, Sydney, Australia

I first met Bill in 1994, just as I was about to start my

Ph.D. working on flat lizards (Platysaurus). I was based

at the Transvaal Museum in Pretoria, and he arrived to

Slice up snakes as part of a project on the ecology of

African snakes with Rick Shine, Jonathan Webb, and

Peter Harlow. Shortly after meeting him, he told me about

the Augrabies flat lizard system, which ended up being

the subject of my Ph.D. and many happy field seasons.

I owed him a huge debt, without realising it at the time!

And as it turned out, our discussions about flat lizards led

to a collaboration that continued until his death.

Everyone that meets Bill is immediately struck by

how warm and caring he is. It’s hard to describe, but

he had a personality that immediately drew you in. And

I think that’s why he had such an impact on so many

people. He was particularly giving and helpful to young

aspiring herpetologists, and I very much appreciated his

friendship and advice as a young Ph.D. student fresh on

the herpetological scene in South Africa. A few years

into my Ph.D., he invited me on a field trip to a remote

area of northern Mozambique to survey the vertebrates

of the Moebase region, the site of a proposed titanium

mine (sadly). I really appreciated this gesture, because

he could have invited any number of far more qualified

people! His son Tom was also on the trip, to survey

birds. Little did I know that this would become such

a memorable trip, and that I would have experiences I

still talk about to this day. There is nothing like a field

trip to really get to know someone, and that trip forged a

lifelong friendship. With Bill, there was never a shortage

of stimulating conversation on a wide range of topics

beyond herpetology. His love for natural history was

Macquarie

Amphib. Reptile Conserv.

XXVviii

infectious. The only thing Bill spoke about with more

passion was his wife Donvé. While on that same field

trip to Mozambique, Bill set out to find a clay pot that

was representative of the region, to take back to Donveé.

I should mention that Donvé is an award winning potter,

so this was the perfect gift! Our fixer couldn’t quite

understand what a westerner would want with a clay pot,

but we examined quite a few, before buying one from

a surprised local villager. Actually, I also acquired one

which has survived multiple moves in South Africa and

a final move to Australia. (How could I not buy one after

hearing Bill wax on about Donveé and her pottery!)

Bill was larger than life and made a huge impact on

African herpetology. It’s hard to accept that he’s gone, but

he will never be forgotten. He will certainly be missed by

many. I am currently working on finishing a phylogeny

and revision of the Platysaurus with Scott Keogh

and Mitzy Pepper, a project that Bill and the late Don

Broadley were both involved in, and he will certainly be

in our thoughts as we put together the final touches.

Fig. 39. Bill and his son Tom while passing through a village

during our 1997 Mozambique trip (scanned from a slide). To

this day, that field trip ranks as one of my most memorable

(Photo: Martin Whiting).

egeerie BA ee io ee at ie

Fig. 40. Bill in action during our 1997 Mozambique trip

[scanned from slides] (Photo: Martin Whiting).

September 2019 | Volume 13 | Number 2 | e186

Literature Cited

Conradie et al.

Li Vigni F (Editor). 2013. A Life for Reptiles and Amphibians,

Volume 1. Chimaira, Frankfurt, Germany. 495 p.

Branch B. 2008. Tortoises, Terrapins & Turtles of Africa. | Shirley MH, Carr AN, Nestler JH, Vliet KA, Brochu

Struik Publishers, Cape Town, South Africa. 128 p. CA. 2018. Systematic revision of the living African

Burger M. 1988. Geographical distribution: Gerrhosaurus Slender-snouted Crocodiles (Mecistops Gray, 1844).

typicus. Journal of the Herpetological Association of Zootaxa 4504(2): 151-193.

Africa 35: 36.

Amphib. Reptile Conserv.

Spawls S, Branch B. 1995. The Dangerous Snakes of

Africa. Southern Book Publishers, Halfway House,

Johannesburg, South Africa. 192 p.

Werner Conradie holds a Masters in Environmental Science (M.Env.Sc.) and has 12 years

of experience with the southern African herpetofauna. His main research interests focus on

the taxonomy, conservation, and ecology of amphibians and reptiles. Werner has published

numerous principal and collaborative scientific papers, and has served on a number of

conservation and scientific panels, including the Southern African Reptile and Amphibian

Relisting Committees. Werner has undertaken research expeditions to various countries

including Angola, Botswana, Lesotho, Malawi, Mozambique, Namibia, South Africa, Zambia,

and Zimbabwe. He is currently the Curator of Herpetology at the Port Elizabeth Museum

(Bayworld), South Africa.

Michael L. Grieneisen spent much of his childhood searching for and observing herps in the

Appalachian Mountains of Pennsylvania, USA. He obtained a B.S. in Biology and Chemistry

from Shippensburg University in Pennsylvania, and a Ph.D. in Biology from University of

North Carolina, Chapel Hill, on a National Science Foundation graduate fellowship. Mike’s

Ph.D. and post-doc work (at University of Nevada, Reno) investigated the hormones that turn

caterpillars into butterflies. Over the past 12 years at University of California, Davis, Mike has

authored journal articles in fields as diverse as nanotechnology, climate change, biodiversity,

scientometrics, environmental science, and reduced-risk pest control practices in California.

Mike is a freelance editor, co-editor of Amphibian & Reptile Conservation, and he is compiling

metadata for the theses and dissertations on amphibians and reptiles produced worldwide.

The compilation currently includes over 54,000 theses, completed from 1803 to the present at

institutions in well over 100 countries, and is expected to be made available sometime in 2020.

He also has an extensive collection of world banknotes which feature herps in the design.

Craig L. Hassapakis is the Founder, Co-editor, and Publisher of the journal Amphibian &

Reptile Conservation (official journal website: amphibian-reptile-conservation.org), which

was founded in 1996, and a former editor of FrogLog (www.amphibians.org/froglog/). Craig

has been an instructor (first grade through college), non-profit and governmental volunteer at

Public Library of Science (PLoS), Co-group Facilitator, Genome Resources Working Group,

IUCN/SSC Amphibian Specialist Group (ASG), and is a member of the IUCN/SSC Amphibian

Specialist Group. His interests include biodiversity, evolution, systematics, phylogenetics,

taxonomy, conservation, and behavior of amphibians and reptiles. Craig is instrumental in

developing and establishing “Amphibia Bank: A genome resource cryobank and network for

amphibian species worldwide.” His professional memberships include: Society for the Study

of Amphibian and Reptiles (SSAR), Herpetologists’ League (HL), International Society for

Biological and Environmental Repositories (ISBER), and International Society for the History

and Bibliography of Herpetology (ISHBH).

XXix September 2019 | Volume 13 | Number 2 | e186

Official journal website:

amphibian-reptile-conservation.org

Amphibian & Reptile Conservation

13(2) [Special Section]: 1-28 (e181).

A herpetological survey of western Zambia

Gabriela B. Bittencourt-Silva

Department of Life Sciences, Natural History Museum, London, SW7 5BD, UNITED KINGDOM

Abstract.—A list of 60 species of amphibians and reptiles found during a six-week survey in western Zambia

is presented. Two species of amphibians are newly reported for Zambia: Amietia chapini and an undescribed

species of Tomopterna, previously known to occur in the Democratic Republic of Congo and in Namibia,

respectively. Some of the material collected could not be confidently identified to species level because of

the taxonomic complexity and uncertainty of some groups (e.g., Phrynobatrachus, Ptychadena), even with

the use of DNA barcoding. This list is a small contribution to the growing knowledge of Zambian and African

herpetology.

Keywords. Amphibians, barcode, checklist, reptiles, Southern Africa, undescribed species

Citation: Bittencourt-Silva GB. 2019. Herpetological survey of western Zambia. Amphibian & Reptile Conservation 13(2) [Special Section]: 1-28 (e181).

tribution 4.0 International (CC BY 4.0): https://creativecommons.org/licenses/by/4.0/], which permits unrestricted use, distribution, and reproduction in

any medium, provided the original author and source are credited. The official and authorized publication credit sources, which will be duly enforced,

are as follows: official journal title Amphibian & Reptile Conservation; official journal website: amphibian-reptile-conservation.org.

Submitted: 2 September 2018; Accepted: 4 March 2019; Published: 6 August 2019

Introduction

Zambia is a landlocked southern African country

considered part of the Zambesiaca area, which also

includes Botswana, Malawi, Mozambique, parts of

Namibia (Caprivi), and Zimbabwe (Poynton and

Broadley 1991). Zambia is located on the main central

African plateau where elevations range from 1,200 m

to 1,500 m and the vegetation 1s dominated by miombo

woodland (Phiri 2005).

Very little has been published on the herpetofauna of

Zambia since the first then comprehensive reports from the

early 1900’s (see reviews in Haagner etal. 2000; Pietersen et

al. 2017). Poynton and Broadley’s compendium Amphibia

Zambesiaca (Poynton and Broadley 1985a,b, 1987, 1988,

1991) and Channing (2001) reported on the distribution of

Zambian amphibians, while Broadley (1971) presented an

initial treatise of the reptiles and amphibians of Zambia,

and Broadley et al. (2003) provided an updated atlas and

field guide to the snakes of Zambia. Similarly, Haagner et

al. (2000) and more recently Pietersen et al. (2017) have

made important contributions to Zambian herpetology.

Broadley (1991) presents a comprehensive list of

reptiles and amphibians from the Mwinilunga District,

northwestern Zambia, including records from museum

collections dating from 1957. However, except from the

extreme north-west (Hillwood Farm), the herpetofauna of

western Zambia remains very poorly studied, with only a

few regional checklists (e.g., Broadley 1991; Pietersen et

al. 2017).

Correspondence. g. bittencourt@nhm.ac.uk

Amphib. Reptile Conserv.

Currently, there are 189 species of reptiles recorded

for Zambia according to The Reptile Database (Uetz

et al. 2018) and 181 (two crocodile, 10 chelonian, 78

lizard, and 91 snake species) according to Pietersen

et al. (2017). The number of amphibian species varies

substantially according to different sources. Pietersen et

al. (2017) report 86 species of amphibians for Zambia,

while AmphibiaWeb (2018) reports 87 species, and

a search in the Amphibian Species of the World 6.0

database (ASW; Frost 2018) returns 104 species. This

disparity between databases is possibly due to the fact

that the ASW includes non-confirmed occurrences. An

example is the caecilian Boulengerula, which is expected

to occur in Zambia based on its known distribution but is

as yet unreported there.

Herein I present a checklist of species collected during

a six-week herpetological survey in western Zambia.

Materials and Methods

Study site and sampling

The survey was carried out in April and May 2014

encompassing protected as well as non-protected areas

of western Zambia (Fig. 1, Table 1). Figure 2 shows

the different vegetation types surveyed, comprising

miombo woodlands (dominated by Brachystegia spp.),

dry evergreen forests (dominated by Cryptosepalum sp.),

riverine forests (mushito) and grassy wetlands (dambo).

During the whole survey period there were only four

August 2019 | Volume 13 | Number 2 | e181

Herpetological survey of western Zambia

sinker mr. 10

Elevation (m)

M200 °° -15

Mi 430

|_| 660

890

1120

[) 1350

{| 1580

|_| 1810

|_| 2040

[__] 2270

[| 2500

Fig. 1. Map of Zambia showing sites surveyed for herpetofauna. The star indicates the capital (Lusaka).

days of rain and average temperatures were 30 °C during

the day and 15 °C at night. The main sampling methods

were acoustic and visual encounter surveys (diurnal

and nocturnal). Entomologists participating in the

expedition opportunistically collected some specimens

with the use of sweep nets and small pitfall traps (500

ml cups). All specimens collected were euthanized with

20% benzocaine (applied on the skin or in the mouth).

Samples of thigh muscles were taken and stored in

absolute ethanol before the specimens were fixed in 10%

formalin and transferred to 70% industrial methylated

spirit for long-term storage. All specimens are deposited

in the herpetological collection of the Natural History

Museum in London, United Kingdom (see Appendix 1).

Species identification

Identification keys for Amphibia (Channing 2001;

Poynton and Broadley 1985a) and Reptilia (Branch

1998; Broadley 1971: Broadley et al. 2003) were used to

Table 1. Localities in Zambia surveyed during this study. Protected areas are indicated by shaded green. NP: National Park; HQ:

Headquarters.

Locality District

Chavuma FR Chavuma

Hillwood Farm Ikelenge

Itezhi-Tezhi, Kafue NP Itezhi-Tezhi

Lukwakwa Kabompo

Livingstone, Maramba Lodge Livingstone

Mayukuyuku, Kafue NP Mumbwa

Nanzila Plains, Kafue NP Itezhi-Tezhi

Ngonye Falls Camp Shangombo

Nkwaji Mwinilunga

Sioma Ngwezi NP Shangombo

Sioma Ngwezi NP (HQ) Shangombo

Amphib. Reptile Conserv.

Longitude Latitude Elevation (m)

-13.07006 22.92880 1070

-11.26316 24.32782 1400

-15.77340 26.01151 1040

-12.66084 24.43697 1150

-17.89120 25.85821 900

-14.91533 26.06311 1010

-16.28138 25.91676 1030

-16.66139 23.57280 930

-11.56567 24.52605 1300

-16.89873 23.59847 1010

-16.66953 23.56743 1000

August 2019 | Volume 13 | Number 2 | e181

Bittencourt-Silva

Fig. 2. Habitats surveyed in western Zambia. (A) Dambo and Cryptosepalum dry forest in Lukwakwa, (B) Dambo in Nanzila Plains,

assist with the identification of specimens. Some species

identifications presented here are tentative because

some groups have complex and difficult taxonomies

(e.g., Hyperolius, Hemisus, Phrynobatrachus, and

Ptychadena). Most samples were barcoded to help

species identification (see details below). Despite its

known limitations (e.g., Deichmann et al. 2017; Hebert

and Gregory 2005; Meier et al. 2006), DNA barcoding

is generally, and sometimes very, helpful. The Basic

Local Alignment Search Tool (BLAST; Altschul et al.

1990) was used to search the GenBank repository and

identify the closest matches for each sample. As there are

not many 16S rRNA sequences of Zambian reptiles and

amphibians openly available for comparison, percentage

of sequence similarity presented here should be

interpreted with caution and while taking the possibility

of geographic isolation or isolation by distance into

account. Private databases were also used for sequence

comparisons (D. Portik and B. Zimkus). Snakes were

identified primarily using morphological characters.

Genetic analysis

Given the large amount of 16S rRNA sequence data

available in GenBank for African amphibians and

reptiles, this gene was selected for DNA barcoding. Total

genomic DNA was extracted using a Qiagen DNeasy

kit (Venlo, Netherlands) following the manufacturer’s

protocol for purification of total DNA from animal

tissues. A fragment (ca. 500 bp) of the 16S rRNA

Amphib. Reptile Conserv.

mitochondrial gene was amplified using the primers

16S H3062 (CCGGTTTGAACTCAGATCA) and 16SB

FROG (CGCCTGTTACCAAAAACAT) [modified from

Palumbi et al. 1991]. Polymerase chain reaction (PCR)

was performed using Illustra PuReTaq Ready-To-Go

PCR Beads (GE Healthcare Life Sciences) for 35 cycles

of 1 minute with annealing temperature at 51 °C. Single

strand sequencing reactions and electrophoresis were

carried out by the molecular lab team at the Natural

History Museum in London, United Kingdom. All

sequences generated are available in GenBank under

accession numbers MK464267—MK464483.

DNA sequences were trimmed in Genelous v.7

(Kearse et al. 2012) with a maximum of low-quality

bases of 20. Uncorrected pairwise distances (p-distances)

of the 16S sequences were calculated for some groups

in PAUP* (Swofford 2001). For Phrynobatrachus,

a maximum likelihood (ML) analysis with non-

parametric bootstrapping was carried out with RAxML

v.8.2 (Stamatakis 2014). The alignment was generated

in Geneious using the Auto algorithm of MAFFT v.7

(Katoh et al. 2002), inspected visually and poorly aligned

regions were eliminated using the GBlocks Server

v.091b (Castresana 2000). Evolutionary models were

evaluated using Automated Model Selection (using a

Neighbor Joining tree) in PAUP*. The best fitting model

(GTR + GAMMA) was selected according to the Akaike

Information Criteria (AIC). Tomopterna marmorata was

used for rooting.

August 2019 | Volume 13 | Number 2 | e181

Herpetological survey of western Zambia

Results

A total of 40 species of amphibians (anurans) belonging

to nine families and 13 genera, and 20 species of reptiles

from nine families and 17 genera (14 lizards, five snakes,

one tortoise) were recorded during this survey (Appendix

1). Among the localities surveyed, Hillwood Farm had

the highest species diversity (n=23), followed by Nkwaji

(n=15). Different from all other areas surveyed, which

are characterized by a combination of miombo woodland,

dambo and/or dry forest, both localities mentioned above

have riverine or swamp forest, locally known as mushitos

(Fig. 2).

Species accounts

All collected specimens and their respective vouchers are

listed below. Voucher numbers in bold refer to specimens

identified solely based on morphology (i.e., no tissue

sample available).

Amphibia

Order Anura

Arthroleptidae

Arthroleptis stenodactylus Pfeffer, 1893

Shovel-footed Squeaker

Material. LUKWAKWA: BMNH 2018.5826, BMNH

2018.5827 (Fig. 3A), BMNH 2018.5828-29; NKWAJI:

BMNH 2018.5830. Comments: Found in leaf litter in

Cryptosepalum forest, in dambo and at edges of mushito.

Arthroleptis stenodactylus as currently understood is

widely distributed from Angola to Tanzania, and from

Kenya to South Africa. This taxon clearly represents a

species complex, possibly two ecologically distinct forms

(see comments in Pickersgill 2007). All specimens listed

above have white venters without any dark markings,

large inner metatarsal tubercles and a dark line on each

side running from the snout over the tympanum to the

shoulder. Sequence similarity with A. stenodactylus from

Malawi is 98% (GenBank accession numbers FJ51098—

99).

Arthroleptis xenochirus Boulenger, 1905

Plain Squeaker

Material. LUKWAKWA: BMNH 2018.5811 (Fig.

3B), BMNH 2018.5812—13; HILLWOOD FARM:

BMNH 2018.5814—20; NKWAJI: BMNH 2018.5821-—

25. Comments: Specimens were found in leaf litter in

Cryptosepalum and mushito. All specimens have a very

small tympanum and relatively large inner metatarsal

tubercle (when compared to A. xenodactyloides). The

closest match on GenBank (94%) is to A. xenodactyloides

from Malawi (FJ151103). There is no sequence of A.

xenochirus available for comparison.

Amphib. Reptile Conserv.

Bufonidae

Schismaderma carens (Smith, 1848)

Red Toad

Material. CHAVUMA FR: BMNH 2018.5729; ITEZHI-

TEZHI: BMNH 2018.5724—28 (Fig. 3C). Comments:

Specimens were found in miombo woodland. This

species 18 widely distributed in Zambia. The BLAST

result show 100% sequence similarity with S. carens

from South Africa (KF665176, AF220913).

Sclerophrys gutturalis (Power, 1927)

Guttural Toad

Material. CHAVUMA FR: BMNH_ 2018.5703;

LIVINGSTONE: BMNH 2018.5702; LUKWAKWA:

BMNH 2018.5705—06, BMNH 2018.5707,

MAYUKUYUKU: BMNH_ 2018.5701 (Fig. 3D);

NKWAJI: BMNH 2018.5704. Comments: This species

is found in miombo and Cryptosepalum forest. These

specimens lack the typical red infusions on their thighs,

although this could be due to preservation. Specimen

identification was confirmed using DNA barcoding

(100% sequence similarity with AF220876, from

Botswana).

Sclerophrys lemairii (Boulenger, 1901)

Yellow Swamp Toad

Material. HILLWOOD FARM: BMNH 2018.5723;

LUKWAKWA: BMNH_ 2018.5715—22 (Fig. 3E).

Comments: One male was found in a pond at night

and eight individuals were found in dambo near the

Cryptosepalum forest. About six males were calling

during the day and two couples were observed in

amplexus. The species exhibits dynamic sexual

dichromatism, where males undergo a temporary color

change (from dark green to bright yellow), depending

on the breeding period; females are reddish, especially

the parotid glands (Bittencourt-Silva 2014; Conradie and

Bills 2017).

Sclerophrys pusilla (Mertens, 1937)

Southern Flat-backed Toad

Material. ITEZHI-TEZHI: BMNH _ 2018.5709-10;

MAYUKUYUKU: BMNH_ 2018.5708 (Fig. 3F);

NKWAJI: BMNH — 2018.5711-12. Comments:

Specimens identification were confirmed using DNA

barcoding (100% sequence similarity) and non-

morphometric morphological characters following

Poynton et al. (2016). Specimens were found in miombo

woodland.

Hemisotidae

Hemisus cf. guineensis Cope, 1865

Guinea Snout-burrower

Material. HILLWOOD FARM: BMNH _ 2018.5801

(juvenile); LUKWAKWA: BMNH 2018.5800 (Fig. 3G);

August 2019 | Volume 13 | Number 2 | e181

Bittencourt-Silva

“te a . »

Fig. 3. Amphibians of western Zambia. (A) Arthroleptis stenodactylus (BMNH 2018.5827), (B) Arthroleptis xenochirus (BMNH

2018.5811), (C) Schismaderma carens (BMNH 2018.5728), (D) Sclerophrys gutturalis (BMNH 2018.5701), (E) Sclerophrys

lemairii, (F) Sclerophrys pusilla (BMNH 2018.5708), (G) Hemisus cf. guineensis (BMNH 2018.5800), (H) Hemisus cf. guineensis

(BMNH 2018.5799), (1) Hemisus marmoratus (BMNH 2018.5713), (J) Hemisus marmoratus (BMNH 2018.5714), (KX) Hyperolius

dartevellei (BMNH 2018.5681), (L) Hyperolius major (BMNH 2018.5675), (M) Hyperolius marginatus (BMNH 2018.5667),

(N) Hyperolius nasicus (BMNH 2018.5666), (O) Hyperolius parallelus (BMNH 2018.5689), (P) Hyperolius parallelus (BMNH

2018.5697), (Q) Hyperolius quinquevittatus, (R) Kassina senegalensis.

Amphib. Reptile Conserv. August 2019 | Volume 13 | Number 2 | e181

Herpetological survey of western Zambia

SIOMA NGWEZI NP: BMNH 2018.5799 (juvenile;

Fig. 3H). Comments: One juvenile (BMNH 2018.5801)

was found dead in a pitfall trap set for dung beetles near

mushito. One large individual (snout-vent length 42.2

mm) from Lukwakwa was found buried in sandy soil

under a log between Cryptosepalum forest and dambo.

The specimen from Sioma Ngwezi is a juvenile. The

BLAST search shows 95% sequence similarity with H.

guineensis from the Republic of the Congo (K Y080117—

19). According to Channing and Broadley (2002),

Hemisus barotseensis, which is endemic to western

Zambia, differs from H. guineesis and H. marmoratus

in body proportions. The presence of a bright yellow

vertebral stripe and small yellow spots on the back agree

with the description of H. barotseensis and it is possible

that these specimens are that species. If confirmed, this

would represent an extension both north and south from

its current known range.

Hemisus marmoratus (Peters, 1854)

Marbled Snout-burrower

Material. MAYUKUYUKU: BMNH 2018.5713 (Fig.

31); SIOMA NGWEZI NP: BMNH 2018.5714 (Fig. 3J).

Comments: Juvenile individuals found at night in sandy

soil in miombo woodland. Hemisus marmoratus is widely

distributed in sub-Saharan Africa, excluding rainforest,

and it mainly inhabits savannahs but can also be found

in gallery forests. The BLAST search shows the closest

match on GenBank (94%) is H. marmoratus (AY 531831).

Table 2 shows the p-distance for both species of Hemisus

presented here and highlights a limitation of the use of

this measure for species delimitation.

Hyperoliidae

Hyperolius dartevellei Laurent, 1943

Dartevelle's Reed Frog

Material. CHAVUMA FR: BMNH 2018.5681 (Fig.

3K); HILLWOOD FARM: BMNH _ 2018.5683-—84;

LUKWAKWA: BMNH — 2018.5682. Comments:

Specimens found in miombo woodland and edge of

mushito basking on vegetation during the day. According

to Channing et al. (2013) the snout profile of H.

dartevellei is truncated instead of shark-like or rounded,

but none of the specimens listed above have truncated

snouts. However, the BLAST results show 98-99%

sequence similarity with samples of H. dartevellei from

Ikelenge, north-western Zambia (JQ863650, JQ863653,

JQ863673, JQ863676—-78, JQ863704, JQ863708,

JQ863718, JQ863750, JQ863753—-54, JQ863756—-S9,

KY080197, KY080199).

Hyperolius kachalolae Schiotz, 1975

Kachalola Reed Frog

Material. HILLWOOD FARM: BMNH 2018.5676,

BMNH 2018.5677-80. Comments: Juvenile specimens

collected during the day on vegetation near a stream in

Amphib. Reptile Conserv.

mushito forest. In life, the overall coloration was green

with a faint canthal and dorsolateral line, consisting of

small spots. The green color faded after preservation,

although the line is still visible. This agrees with the

description provided by Schietz (1975). The sequenced

individual shows 98% similarity with H. kachalolae

from northern Zambia (D. Portik, pers. comm. ).

Hyperolius major Laurent, 1954

Material. HILLWOOD FARM: BMNH 2018.5675 (Fig.

3L). Comments: One male found on top of a leaf (1.5 m

from the ground) calling at night. Schietz (1999) reports

this as a savannah species from north-western Zambia

and eastern Democratic Republic of the Congo (DRC),

however, this specimen was found in a forest patch

(mushito). The closest matches from DNA barcoding

(97%) are H. kuligae and H. langi (D. Portik, pers.

comm. ). Schiotz (1999) states that these species found in

west and central Africa are very similar in morphology

and dorsal pattern, and may be conspecific. In contrast,

Kohler et al. (2005) treat H. kuligae as a western and H.

langi as an eastern Central African form. However, the

color pattern (especially the post orbital marking) differs

substantially from Laurent’s description of the type

material of H. /angi, whereas this specimen agrees with

the morphological description of H. major provided by

Schiotz (1999). There is no DNA sequence of H. major

available for comparison.

Hyperolius marginatus Peters, 1854

Margined Reed Frog

Material. LUKWAKWA: BMNH 2018.5667 (male;

Fig. 3M); NANZILA PLAINS BMNH 2018.5668,

BMNH 2018.5674 (juveniles). Comments: Specimens

found near ponds in miombo woodland. There is 100%

sequence similarity with H. marginatus from Zambia (D.

Portik, pers. comm. ).

Hyperolius nasicus Laurent, 1943

Pointed Long Reed Frog

Material. NANZILA PLAINS BMNH _ 2018.5666

(Fig. 3N). Comments: This single specimen was

collected during the day while resting on vegetation ca.

1 m above the ground in dambo. The specimen fits the

morphological description of the species provided by

Channing et al. (2013): when viewed in profile the snout

has a shark-like tip, and the first, third, and fifth toes have

one phalanx free (or nearly free) of webbing (see Fig. 4),

distinguishing it from all the other species. The closest

match on GenBank (98-99% sequence similarity) is

Hyperolius inyangae (JQ863674, JQ863683-—84), which

is only known from the Eastern Highlands of Zimbabwe

and Malawi (Channing et al. 2013). Although Channing

et al. (2013) provide accession numbers for the genetic

material of H. nasicus, no sequences could be found on

GenBank under this species name. This could be due

to sequence mislabelling and the accession numbers

August 2019 | Volume 13 | Number 2 | e181

Bittencourt-Silva

Fig. 4. Details of head profile and webbing of Hyperolius

nasicus (BMNH 2018.5666). (A) Profile of head, (B) webbing

of right foot, and (C) schematic representation of webbing.

Scale bar represents 1 mm.

associated with H. invangae may actually be from H.

nasicus.

Hyperolius parallelus Giinther, 1858

Angolan Reed Frog

Material. HILLWOOD FARM: BMNH 2018.5687-88,

BMNH 2018.5689 (Fig. 30), BMNH 2018.5690-94,

BMNH 2018.5695; NKWAJI: BMNH_ 2018.5696,

BMNH 2018.5697 (Fig. 3P), BMNH 2018.5698—5700.

Comments: Specimens found near ponds in miombo

woodland. These specimens show a color variation

similar to the alborufus group (see Schiotz 1999).

Hyperolius parallelus is a taxonomically complex group

due to its considerable color polymorphism. According

to the BLAST search, the closest match (98% sequence

similarity with JQ513623, JQ513626, and JQ513625)

is H. angolensis from Angola (see Conradie et al. 2012;

Frost 2018).

Hyperolius quinquevittatus Bocage, 1866

Five-striped Reed Frog

Material. NKWAJI: BMNH 2018.5685—86 (Fig. 3Q).

Comments: Juveniles collected during the day while

resting on vegetation in mushito. BLAST results show

99% sequence similarity with H. quinquevittatus from

Ikelenge, north-western Zambia (GenBank accession

number JQ863752).

Kassina senegalensis (Dumeéril and Bibron, 1841)

Bubbling Kassina

Material. CHAVUMA FR: BMNH = 2018.5810;

HILLWOOD FARM: BMNH 2018.5802-03 (Fig. 3R);

NKWAJI: BMNH 2018.5804; SIOMA NGWEZI NP:

BMNH 2018.5805—09. Comments: Specimens found in

Table 2. Uncorrected pairwise distances (p-distances) for a fragment of the 16S rRNA gene for Hemisus spp. Distances between

conspecific populations are inside boxes.

Taxon ID 1 2 3 4 5

1 Hemisus marmoratus AY326070

2 Hemisus marmoratus DQ283430

3 Hemisus marmoratus AY531831

4 Hemisus marmoratus KY 176997

5 Hemisus marmoratus AY948749

6 Hemisus marmoratus KX492610

7 Hemisus marmoratus KM509138

8 Hemisus marmoratus KY 176998

9 Hemisus marmoratus BMNH 2018.5713

10 Hemisus marmoratus BMNH 2018.5714

11 Hemisus guineensis KY080117

12 Hemisus guineensis KY080118

13. Hemisus guineensis KY080119

14 Hemisus guineensis KY080120

15 Hemisus cf guineensis BMNH 2018.5799

16 Hemisus cf guineensis BMNH 2018.5800

17. -Hemisus cf guineensis BMNH 2018.5801

Amphib. Reptile Conserv.

August 2019 | Volume 13 | Number 2 | e181

Herpetological survey

100

Namibia (GU457565)

100 | Lindi, Tanzania (KY177049)

Beira, Mozambique (DQ022361)

Tomopterna marmorata: Zambia (BMNH_2018.5792)

0.05

Ethiopia (FJ829292)

Ethiopia (FJ829296)

Ethiopia (FJ829295)

Ethiopia (FJ829297)

Uganda (FJ829298)

Ghana (FJ769126)

Ghana (FJ769127)

Céte d'Ivoire (GU457566)

Guinea (EU718726)

Sioma Ngwezi NP (BMNH_2018.5864)

Sioma Ngwezi NP (BMNH_2018.5865)

Ngonye Falls Camp (BMNH_2018.5836)

Ngonye Falls Camp (BMNH_2018.5855)

Mtunzini, South Africa (DQ019605)*

South Africa (DQ347303)

of western Zambia

Itezhi-Tezhi, Kafue NP (BMNH_2018.5848)

Itezhi-Tezhi, Kafue NP (BMNH_2018.5849)

Itezhi-Tezhi, Kafue NP (BMNH_2018.5850)

Nanzila Plains, Kafue NP (BMNH_2018.5851)

Nanzila Plains, Kafue NP (BMNH_2018.5852)

Mayukuyuku,

Maramba Lodge, Livingnstone (BMNH_2018.5853)

Maramba Lodge, Livingnstone (BMNH_2018.5854)

Lake Malawi Park, Malawi (FJ889462)

Kakamega Forest, Kenya (FJ889464)

Kakamega Forest, Kenya (FJ889463)

Tatanda Village, Tanzania (DQ283414)

Northern Territory, Rwanda (FJ829290)

Northern Territory, Rwanda (FJ829291)

Kigoma Region, Tanzania (FJ829299)

Tanzania (FJ829289)

lringa, Tanzania (FJ829293)

Kakamega Forest, Kenya (FJ889463)

Kakamega Forest, Kenya (FJ889464)

Kigoma Region, Tanzania (FJ829300)

Mangochi District, Malawi (FJ889462)

Niassa Game Reserve, Mozambique (FJ829303)

Niassa Game Reserve, Mozambique (FJ829301)

Niassa Game Reserve, Mozambique (FJ829302)

Morogoro Region, Tanzania (FJ829294)

Kafue NP (BMNH_2018.5837)

Kafue NP (BMNH_2018.5838)

Kafue NP (BMNH_2018.5839)

Kafue NP (BMNH_2018.5840)

Kafue NP (BMNH_2018.5841)

Kafue NP (BMNH_2018.5842)

Kafue NP (BMNH_2018.5843)

Haplotype

Group A**

Haplotype

Group B**

| Sp.2

| Phrynobatrachus natalensis

Nkwaji (BMNH_2018.5863)

Nkwaji (BMNH_2018.5858)

Nkwaji (BMNH_2018.5860)

Hillwood Farm (BMNH_2018.5866)

Nkwaji (BMNH_2018.5856)

Nkwaji (BMNH_2018.5859)

Nkwaji (BMNH_2018.5871)

Nkwaji (BMNH_2018.5872)

Nkwaji (BMNH_2018.5869)

Nkwaji (BMNH_2018.5868)

Hillwood Farm (BMNH_2018.5867)

Nkwaji (BMNH_2018.5870)

Sp.3

Fig. 5. Maximum likelihood phylogenetic tree inferred from nucleotide sequence data from mitochondrial 16S rRNA of

Phrynobatrachus natalensis. Numbers above branches are non-parametric bootstrap support values. Specimen vouchers or GenBank

accession numbers are shown in parentheses. Colored polygons highlight the clades comprising specimens from this study. (*)

Nearest sample from type locality of Phrynobatrachus natalensis; (**) Haplotype groups A and B in Zimkus and Schick (2010).

miombo woodland near temporary ponds. All sequences

closely match K. senegalensis (98%, GenBank accession

number AF215445).

Phrynobatrachidae (see Table 3 for inter- and intra-

specific p-distances; Fig. 5 shows maximum likelihood

tree for this group)

Phrynobatrachus cf. parvulus (Boulenger, 1905)

Small Puddle Frog

Material. HILLWOOD FARM: BMNH 2018.5873-78;

LUKWAKWA: BMNH 2018.5889 (Fig. 5A); NKWAJI:

BMNH 2018.5882 (juvenile), BMNH 2018.5879-80,

BMNH 2018.5883—88. Comments: All specimens were

found during the day in dambo. Males have a dark throat

(BMNH 2018.5882, BMNH 2018.5886—87). While the

specimens of P. mababiensis listed below have the venter

immaculate (creamy), these specimens have the venter

white with dark speckles. Additionally, these specimens

show a more well-defined band on the thigh (which

runs from knee to knee) and, in most specimens, a light

Amphib. Reptile Conserv.

line runs along the tibia-fibula and thigh (parallel to the

band) and joins a vertebral line above the vent (see Fig.

6A). According to Du Preez and Carruthers (2017), the

presence of the latter feature distinguishes P. parvulus

from P. mababiensis. However, this character is present

on both species and therefore cannot be used to separate

them (see Poynton and Broadley 1985a. Pietersen et al.

(2017) report P. parvulus for Ngonye Falls, approximately

25 km from Sioma Ngwesi NP, but unfortunately there

is no voucher specimen. Poynton and Broadley (1985a)

and Marques et al. (2018) provide discussions of the

literature on the identifications of P. mababiensis and

P. parvulus. The barcode is very inconclusive given

that the closest hits on GenBank (92-95%) include

samples of an unidentified species of Phrynobatrachus

from Gabon (KP247505), one from the Republic of the

Congo (KY080354), and P. keniensis (JX564885) and

P. scheffleri (FJ889479), both from Kenya. Poynton

and Broadley (1985a) suggest P. parvulus tends to be

associated more with upland and forest conditions than

August 2019 | Volume 13 | Number 2 | e181

Bittencourt-Silva

Table 3. Uncorrected pairwise distances (p-distances) for the 16S rRNA gene for Phrynobatrachus spp. Distances between

conspecific populations are inside boxes. Dotted-line box indicates P. natalensis group.

1 Phrynobatrachus cf. parvulus BMNH 2018.5873

2 Phrynobatrachus cf. parvulus BMNH 2018.5874 0.01

3 Phrynobatrachus cf. parvulus BMNH 2018.5880 0.04 0.05 -

4 Phrynobatrachus mababiensis FJ889461 0.11 O12 0.12

5 Phrynobatrachus mababiensis BMNH 2018.5831 0.11 O11 0.12

6 Phrynobatrachus mababiensis BMNH 2018.5832 0.11 O11 0.12

7 Phrynobatrachus natalensis DQO019605 0.14 O15 0.14

8 Phrynobatrachus natalensis BMNH 2018.5855 0.14 O15 0.14

9 Phrynobatrachus sp. 1 BMNH 2018.5838 0.16 O17 0.17

10 Phrynobatrachus sp. 1 BMNH 2018.5839 0.16 O17 0.17

11 Phrynobatrachus sp. 3 BMNH 2018.5856 0.15 016 0.15

12 Phrynobatrachus sp. 3 BMNH 2018.5858 0.15 016 0.15

13 Phrynobatrachus sp. 2 BMNH 2018.5864 0.17 O17 0.18

14 Phrynobatrachus sp. 2 BMNH 2018.5865 0.17 O17 0.18

P. mababiensis. The localities where these specimens

were found are all upland and either inside or near forest,

therefore I refer them to P. cf. parvulus.

Phrynobatrachus mababiensis FitzSimons, 1932

Dwarf Puddle Frog

Material. MAYUKUYUKU: BMNH 2018.5831-32,

BMNH_ 2018.5881; NANZILA PLAINS: BMNH

2018.5833; SIOMA NGWEZI NP: BMNH 2018.5834—

35. Comments: All specimens are juveniles and were

found in dambo both during the day and at night.

According to Poynton and Broadley (1985a), it 1s not

easy to distinguish P. mababiensis from P. parvulus based

on external morphology, but they suggest that some

characters usually serve to separate them (1.e., labial and

subtympanic markings, and shape of tarsal tubercle).

The usual well-marked black and white barring on the

upper and lower jaws is rather faint on these specimens.

Zimkus and Schick (2010) suggest that there 1s cryptic

diversity within the P mababiensis group. The closest

match on GenBank is P. mababiensis (FJ889461; 99%

sequence similarity) from eastern Zambia, which belongs

to a population that is sister to the clade containing P.

ukingensis and P. ungujae (see Zimkus and Schick 2010).

Phrynobatrachus natalensis (Smith, 1849)

Snoring Puddle Frog

Material. NGONYE FALLS: BMNH _ 2018.5855.

Comments: The overall external morphology resembles

P. natalensis. The BLAST search shows 98% sequence

similarity to P. natalensis from Mtunzini, South Africa

(DQ347303), which is near Durban, the type locality of

P. natalensis (see Table 3 and Fig. 5).

Phrynobatrachus sp. |

Material. I[TEZHI-TEZHI:

50; LIVINGSTONE:

BMNH

2018.5848—

2018.5853—54;

Amphib. Reptile Conserv.

4 5 6 7 8 9 10 11 12 13 14

0.00

0.00 0.00

0.14 013 013

0.17 0.16 0.16}

017 0.16 0.16}

015 014 0.14} 0.09

0.05 0.08 0.08 7

0.05 0.08 0.08 0.00 |

MAYUKUYUKU: BMNH 2018.5837-38 (Fig. 6B),

BMNH 2018.5839-42 (Fig. 6C), BMNH 2018.5843,

BMNH 2018.5844—-47,; NANZILA PLAINS: BMNH

2018.5851-52. Comments: Specimens were found in

dambos. One juvenile was found ona beach of the Zambezi

River at Ngonye Falls. The closest match on GenBank

(99% sequence similarity) is P. natalensis DQ283414

from Tanzania (see Fig. 5). Zimkus and Schick (2010)

suggest that there are two species of P. natalensis in East

Africa, and these Zambian populations are more similar

to the central and southern populations corresponding to

Haplotype group B. It further corresponds to Zimkus et

al. (2010) P. natalensis Clade E. See further comments

in Discussion.

0.16 015 0153

0.17 0.16 0.16:

0.16 }

0.17 0.16

Phrynobatrachus sp. 2

Material. NGONYE FALLS: BMNH2018.5836; SIOMA

NGWEZI NP: BMNH 2018.5864—-65. Comments:

Morphologically, these specimens resemble P. natalensis

in terms of size, toe webbing and overall color pattern.

However, they present silver/white spots around the vent

(and ventral part of the thigh in BMNH 2018.5836). The

closest match on GenBank is P. natalensis from Tanzania

(95%; DQ283414). Although their range overlaps with

the Southern African geographic zone (populations A

and B) in Zimkus et al. (2010), these populations form

a southern Zambian clade, which is a sister group of the

eastern and western African clades (see Fig. 5). These

findings allude to further cryptic diversity in the group.

Phrynobatrachus sp. 3

Material. HILLWOOD FARM: BMNH 2018.5866—67

(Fig. 6D); NKWAJI: BMNH 2018.5856-57, BMNH

2018.5858-60, BMNH 2018.5861 (Fig. 6E), BMNH

2018.5862-63; BMNH 2018.5868-69 (Fig. 6F),

BMNH _ 2018.5870-72. Comments: All specimens

August 2019 | Volume 13 | Number 2 | e181

Herpetological survey of western Zambia

rs a

An «spi

ae ie i

: Nee

Fig. 6. onan fe western Zaria, (A) re of one (BMNH 2018.5889), (B) Phrynobatrachus natalensis

(BMNH 2018.5838), (C) Phrynobatrachus natalensis (BMNH 2018.5842), (D) Phrynobatrachus sp. 1 (BMNH 2018.5867),

(E) Phrynobatrachus sp. 1 (BMNH 2018.5861), (F) Phrynobatrachus sp. 1 (BMNH 2018.5869), (G) Xenopus poweri (BMNH

2018.5659), (H) Xenopus pygmaeus, (1) Ptychadena anchietae (BMNH 2018.5730), (J) Ptychadena cf. mossambica (BMNH

2018.5759), (IX) Ptychadena porosissima (BMNH 2018.5766), (L) Ptychadena porosissima (BMNH 2018.5769), (M) Ptychadena

porosissima (BMNH 2018.5770), (N) Ptychadena taenioscelis (BMNH 2018.5785), (O) Amietia chapini (BMNH 2018.5664), (P)

Pyxicephalus cf. adspersus (BMNH 2018.5791), (Q) Tomopterna sp. (BMNH 2018.5797), (R) Chiromantis xerampelina (BMNH

2018.5798).

Amphib. Reptile Conserv. 10 August 2019 | Volume 13 | Number 2 | e181

Bittencourt-Silva

are morphologically similar to P. natalensis, however

the BLAST search shows sequence similarity with

P. natalensis from South Africa (GenBank accession

number DQ347303) varying between 90-91% (see Fig.

5). These populations form a northern Zambian clade,

sister to southern, western, and eastern African clades.

Further investigation is needed to resolve the taxonomical

status of this group.

Pipidae

Xenopus poweri Hewitt, 1927

Power’s Clawed Frog

Material. HILLWOOD FARM: BMNH 2018.5654—58;

NKWAJI: BMNH 2018.5659 (Fig. 6G). Comments:

Specimens collected from a pond in miombo area. The

BLAST search shows 99% sequence similarity with X.

poweri from Ikelenge, north-western Zambia (GenBank

accession number KP345253). Furman et al. (2015)

removed this name from the synonymy of X. petersii and

reassigned the West African populations of X. /aevis to

this species.

Xenopus pygmaeus Loumont, 1986

Bouchia Clawed Frog

Material. HILLWOOD FARM: BMNH 2018.5651—52

(Fig. 6H), BMNH 2018.5653. Comments: Species

identification was based on their relatively small size and

the presence of a fourth claw. Specimens were found ina

small pool formed in a car track next to a riverine forest

(mushito). Recently, Wagner et al. (2013) presented

the first record of Xenopus pygmaeus for Zambia,

representing a significant range extension (about 1,300

km). This species belongs to the fraseri subgroup

and it was previously known to have its southernmost

distribution in the northern part of the DRC. The DNA

barcode corroborates the morphological identification

(99% sequence similarity with KF738291).

Ptychadenidae

Ptychadena anchietae (Bocage, 1868)

Anchieta's Ridged Frog

Material. ITEZHI-TEZHI: BMNH_ 2018.5735-36;

MAYUKUYUKU: BMNH 2018.5730-31 (Fig. 61),

BMNH = 2018.5732-34. Comments: Specimens

collected near water in miombo woodland. The closest

match (99%) is P. anchietae (AY517610) from Tanzania.

Ptychadena grandisonae Laurent, 1954

Grandison's Ridged Frog

Material. NK WAJI: BMNH 2018.5737-43. Comments:

Two juveniles and four males found in a pond in miombo.

This series fits the description for P. grandisonae in

Poynton and Broadley (1985a). The results of the

BLAST search show that the closest match (98%) is

Amphib. Reptile Conserv.

Ptychadena sp. (GenBank accession number KF 178892)

from Gabon.

Ptychadena cf. guibei

Material MAYUKUYUKU: BMNH_ § 2018.5764;

SIOMA NGWEZI NP: BMNH 2018.5765. Comments:

Specimens identified following the key in Poynton and

Broadley (1985a). Foot length of BMNH 2018.5765 is

slightly less than half the body length and thus, according

to the key, this specimen would be mossambica or cotti

(i.e. schillukorum). The BLAST result shows 96%

sequence similarity with P. porosissima (KY177058)

from Kenya. There is no sequence of P. guibei available

for comparison.

Ptychadena mapacha Channing, 1993

Mapach Ridged Frog

Material. MAYUKUYUKU: BMNH 2018.5772 (male).

Comments: Specimen collected near water in miombo

woodland. Following Poynton and Broadley (1985a),

this specimen should be assigned to P cotti, now a

synonym of P. schillukorum. However, this specimen

also fits the description of Ptychadena mapacha (not

included in the key), except for the white spots on the

posterior face of the tibia and a thin tibial line that are

present in the holotype (CAS 160535). The closest match

(89%) is P. porosissima (GenBank accession number

KY177058) from Kenya, and the sequence similarity

with P. schillukorum (KY177060) is 82%. There is

no sequence of P. mapacha available for comparison.

Although P. schillukorum is \isted in the AmphibiaWeb

database as occurring in Zambia, there is no reference

to the literature confirming this claim. Pietersen et al.

(2017) provided an unconfirmed record of P. mapacha for

Sioma Ngwesi. This record represents the northernmost

record of this species.

Ptychadena cf. mossambica (Peters, 1854)

Mozambique Ridged Frog

Material. ITEZHI-TEZHI: BMNH 2018.5753, BMNH

2018.5754-57; MAYUKUYUKU: BMNH 2018.5763;

SIOMA NGWEZI NP: BMNH 2018.5758—59 (Fig. 6J),

BMNH 2018.5760, BMNH 2018.5761. Comments:

The key in Poynton and Broadley (1985a) points to

P. mossambica, except for the skin folds that are not

continuous in these specimens. The authors note that

P. mossambica shows an east-west cline in size and

degree of webbing, where material from western

Zambia tends to be smaller (maximum SVL 28.9 mm)

than the series from Mozambique (maximum SVL 52.5

mm). The average SVL of this series is 34 mm. Most

specimens have a pair (sometimes two) of large dark

blotches on the scapular region. The closest match on

GenBank (93-94%) is Ptychadena cf. mossambica from

coastal Tanzania (KY177057). These specimens may

be referrable to PR mapacha Channing, 1993, for which

there is no available sequence data. Ptyvchadena mapacha

August 2019 | Volume 13 | Number 2 | e181

Herpetological survey of western Zambia

has recently been recorded in Ngonye Falls, south-west

Zambia by Pietersen et al. (2017), confirming Channing’s

(2001) prediction about its distribution.

Ptychadena nilotica (Seetzen, 1855)

Nile Grass Frog

Material. LIVINGSTONE: BMNH — 2018.5781;

NANZILA PLAINS: BMNH 2018.5773-76, BMNH

2018.5777—80. Comments: Specimens collected at night

near water in miombo woodland. Sequence similarity

with P. nilotica is 98% with KFO27211 and 99% with

KX836515 from the DRC. For further discussion about

this species see Dehling and Sinsch (2013) and Zimkus

et al. (2016).

Ptychadena obscura (Schmidt and Inger, 1959)

Material. HILLWOOD FARM: BMNH 2018.5766-67

(Fig. 6K), BMNH 2018.5768. Comments: These small

specimens (SVL ranges from 21.8 to 22.8 mm) fit the

description of P. obscura in Poynton and Broadley

(1985a). The specimens have a pair of dark marks on

the scapular region. The results of the BLAST search

show higher similarity (97-98%) to P. broadleyi

(GenBank accession number MH300600-02). This is an

unexpected finding considering that P. broadleyi is only

known to occur in the Mulanje Mountain and the Zomba

Plateau, in Malawi. These specimens from Zambia have

a light triangle on the snout distinguishing them from

P. broadleyi. Hence, the barcoding results should be

interpreted with caution.

Ptychadena oxyrhynchus (Smith, 1849)

Sharp-nosed Grass Frog

Material. HILLWOOD FARM: BMNH 2018.5783;

NANZILA PLAINS: BMNH 2018.5782. Comments:

Specimens collected near water in miombo woodland.

Sequence similarity with P oxyrhynchus from

Kwambonambi, South Africa (GenBank accession

number AF215403) is 99%.

Ptychadena porosissima (Steindachner, 1867)

Three-striped Grass Frog

Material. HILLWOOD FARM: BMNH 2018.5769 (Fig.

6L), BMNH 2018.5770 (Fig. 6M), BMNH 2018.5771.

Comments: This species, common in miombo woodland

near water bodies, was present in large numbers at

Hillwood Farm. Sequence similarity with P. porosissima

(GenBank accession number KF0O27212) from Rwanda

is 98%.

Ptychadena cf. taenioscelis Laurent, 1954

Stripe-legged Grass Frog

Material. HILLWOOD FARM: BMNH 2018.5784—-86

(Fig. 6N). Comments: Adult specimens found in pond.

These specimens have been identified using the key in

Poynton and Broadley (1985a). The closest match on

GenBank (95%) is P. taenioscelis from the Republic of the

Amphib. Reptile Conserv.

Congo (GenBank accession number KY080397). Perret

(1979) assigned records of taenioscelis from west and

central Africa to pumilio Boulenger. There seems to be

some confusion in the literature regarding the taxonomy

of these species, and a review of the group is needed.

Ptychadena upembae (Schmidt and Inger, 1959)

Upemba Ridged Frog

Material. HILLWOOD FARM: BMNH 2018.5750—

52 (last number is a juvenile); NKWAJI: BMNH

2018.574446 (juveniles), BMNH 2018.5747-49 (last

number is a juvenile); SIMA NGWEZI NP: BMNH

2018.5762. Comments: Following the key in Poynton

and Broadley (1985a), this series should be assigned to

Ptychadena upembae. The BLAST search shows that

the closest match (96%) is Ptychadena aff. porosissima

(GenBank accession number DQ525940) from Tanzania,

but it is important to note that there is no sequence of P.

upembae available for comparison.

Pyxicephalidae

Amietia chapini (Noble, 1924)

Chapin's River Frog

Material. HILLWOOD FARM: BMNH 2018.5660-61,

BMNH 2018.5662—63, BMNH 2018.5664 (Fig. 60),

BMNH 2018.5665. Comments: Specimens found near

streams in miombo woodland. The result of the BLAST

search shows that A. chapini is the closest match to

the specimens collected at Hillwood Farm (sequence

similarity with A. chapini of 96-98%). All specimens

have long legs (tibiofibula ~0.6 of snout-vent length),

as noted by Noble (1924). I note that the specimens

described as A. chapini by Channing et al. (2016) differ

from the specimens listed above in coloration — the

latter being darker. If these specimens are confirmed

to be A. chapini, this will be the first record of this

species for Zambia, but their presence in Zambia is

not surprising considering the proximity (ca. 380 km)

between the currently known populations from southern

DRC (Kundelungu National Park) and Hillwood Farm.

Pyxicephalus cf. adspersus Tschudi, 1838

Giant Bullfrog

Material. SIOMA NGWEZI NP: BMNH 2018.5787-

91 (Fig. 6P). Comments: All individuals collected are

juveniles. Hence, identification is tentative and based on

the geographic range of the species. The BLAST shows

95% sequence similarity to Pyxicephalus cf. adspersus

(DQ347304) and P. edulis (DQ022366).

Tomopterna marmorata (Peters, 1854)

Marbled Sand Frog

Material. LIVINGSTONE: BMNH — 2018.5792.

Comments: Juvenile found in a small pond at night.

Skin of dorsum and venter is smooth; venter is pale

August 2019 | Volume 13 | Number 2 | e181

Bittencourt-Silva

Table 4. Uncorrected pairwise distances (p-distances) for the 16S rRNA gene for Zomopterna spp. Distances between conspecific

populations are inside boxes.

1 Tomopterna sp. "Shankara" AY255095 7

Tomopterna sp. BMNH 2018.5793

Tomopterna sp. BMNH 2018.5794

Tomopterna sp. BMNH 2018.5795

Tomopterna sp. BMNH 2018.5796

Tomopterna delalandii DQ283403 0.06 0.06

6 Tomopterna sp. BMNH 2018.5797

8 Tomopterna damarensis KX869909 0.07 0.06

Tomopterna cryptotis JX564898 0.06 0.06

10 Tomopterna marmorata BMNH 2018.5792 0.08 0.08

11. Yomopterna marmorata AY 255084 0.08 0.08

with a few dark markings under the throat; tympanum

indistinguishable; undivided sub-articular tubercle on

first finger. The closest match on GenBank (100%) is

Tomopterna marmorata (AY 255084) from Zambia.

Zomopterna sp.

Material. SIOMA NGWEZI NP: BMNH 2018.5793—

97 (Fig. 6Q). Comments: All individuals are juveniles

and were found at night close to a light-trap set to

collect beetles. The main morphological characters of

these specimens are: dorsal and ventral skin smooth;

immaculate venter; presence of a ridge below tympanum;

tympanum indistinguishable; and undivided sub-articular

tubercle on first finger. The closest match on GenBank

(99%) is Tomopterna sp. “Shankara” (AY255095), an

undescribed species from Namibia (Dawood et al. 2002).

Table 4 shows the p-distances among the closest matches

from GenBank.

Rhacophoridae

Chiromantis xerampelina Peters, 1854

African Grey Treefrog

Material. MAYUKUYUKU: BMNH 2018.5798 (Fig.

6R). Comments: This is a widespread species found in

miombo woodland. One adult individual found at night

on a tree near the campsite.

Reptilia

Order Squamata

Agamidae

Agama armata Peters, 1855

Northern Ground Agama

Material. NGONYE FALLS CAMP: BMNH 2018.2751

(Fig. 7A). Comments: One juvenile found basking

on a log by the Zambezi River. Sequence similarity is

Amphib. Reptile Conserv.

0.06 0.06 0.06 0.06

0.07 0.07 0.07 0.07 0.06 0.05

4 5 6 7 8 i) 10 11

0.05 0.05 0.05 0.05 -

0.02 -

0.06 0.06 0.06 0.06 0.03 0.02 -

0.07 0.07 0.07 0.07 0.06 0.05 0.06 -

00 [aon] -

98% with A. armata ZFMK 84990 (GenBank accession

number GU128447).

Chamaeleonidae

Chamaeleo dilepis Leach, 1819

Flap-necked Chameleon

Material. CHAVUMA FR: BMNH 2018.2755 (Fig.

7B), BMNH 2018.2756. Comments: Specimens were

found in miombo woodland on shrubs above | m. There

are currently seven subspecies in this group and based

on their distributions, these specimens represent C.

dilepis quilensis (Uetz et al. 2018). Sequence similarity

is 99% with Chamaeleo dilepis from Matema, Tanzania

(GenBank accession number AY927272).

Gekkonidae

Hemidactylus mabouia (Moreau de Jonnes, 1818)

Common Tropical House Gecko

Material. I[TEZHI-TEZHI: BMNH 2018.2742, BMNH

2018.2740; NANZILA PLAINS: BMNH 2018.2741.

Comments: Common species found in a variety of

habitats, including heavily degraded ones, though not

found in forests. The closest matches from GenBank are

Hemidactylus mercatorius (AY 863034) and H. mabouia

(AY 863038), both showing 94% sequence similarity.

Lygodactylus chobiensis Fitzsimons, 1932

Chobe Dwarf Gecko

Material. ITEZHI-TEZHI: BMNH 2018.2743 (female;

Fig. 7C), BMNH 2018.2744 (male) Comments:

Specimens found on tree-trunks in miombo. Identification

follows the key provided by Broadley (1971). The

rostral is excluded from the nostril and the male has

dark forward-directed chevron marks on the throat. The

female is yellow and white underneath (Fig. 7D). The

closest match from GenBank (95% sequence similarity)

August 2019 | Volume 13 | Number 2 | e181

Herpetological survey of western Zambia

Fig. 7. Reptiles of western Zambia. (A) Agama armata (BMNH 2018.2751), (B) Chamaeleo dilepis (BMNH 2018.2755), (C}H(D)

Lygodactylus chobiensis (BMNH 2018.2743), (E) Pachydactylus punctatus, (F) Ichnotropis capensis (BMNH 2018.2750), (G)

Meroles squamulosus (BMNH 2018.2753), (H) Trachylepis cf. albopunctata (BMNH 2018.2765), (I) Trachylepis varia (BMNH

2018.2769), (J) Zyphlacontias rohani (BMNH 2018.2761), (IK) Crotaphopeltis hotamboeia (BMNH 2018.2776), (L) Philothamnus

hoplogaster (BMNH 2018.2775), (M) Rhamnophis aethiopissa ituriensis (BMNH 2018.2772), (N) Thelotornis kirtlandii (BMNH

2018.2760), (O) Atractaspis congica (BMNH 2018.2274), (P)-(R) Kinixys spekii.

Amphib. Reptile Conserv. 14 August 2019 | Volume 13 | Number 2 | e181

Bittencourt-Silva

is Lygodactylus chobiensis from Zimbabwe (GenBank

accession number GU593456).

Lygodactylus angolensis Bocage, 1896

Angola Dwarf Gecko

Material. NKWAJI: BMNH 2018.2767 (male), BMNH

2018.2766 (female). Comments: Specimens were

identified following the key provided by Broadley

(1971). In both specimens, the mental has a pair of lateral

clefts, resulting from fusion with a large postmental. The

male has nine preanal pores, which distinguish it from

L. bradfieldi. The closest match from GenBank (90%

sequence similarity) is Lygodactylus sp. from East Africa

(GenBank accession numbers GU593448-50).

Pachydactylus punctatus Peters, 1854

Speckled Thick-toed Gecko

Material. I[TEZHI-TEZHI: BMNH 2018.2757, BMNH

2018.2758 (Fig. 7E), BMNH 2018.2759. Comments:

Specimens found at night in dry leaf litter in miombo

woodland. All specimens have the dorsum covered

with sub-uniform granules. The closest matches from

GenBank (93% sequence similarity) are Pachydactylus

punctatus (AF449120) and P. scherzi (AY123379). As

the latter is only known from Namibia (Bauer and Branch

1995), I assign these specimens to P. punctatus.

Gerrhosauridae

Gerrhosaurus bulsi Laurent, 1954

Laurent’s Plated Lizard

Material. HILLWOOD FARM: BMNH 2018.2754.

Comments: One juvenile collected by the farm scouts in

dry miombo. The BLAST search shows the closest match

(98%) as Gerrhosaurus bulsi from Angola (KF717381).

Broadley (1971, 1991) referred the population of

Gerrhosaurus from Ikelenge (north-western Zambia) to

multilineatus but this was later contested by Haagner et

al. (2000). Bates et al. (2013) consider G. bulsi a valid

species and discuss the taxonomic problems regarding G.

multilineatus.

Lacertidae

Ichnotropis capensis (Smith, 1838)

Cape Rough-scaled Lizard

Material. CHAVUMA FR: BMNH = 2018.2746;

LUKWAKWA: BMNH 2018.2749, BMNH 2018.2747;

NANZILA PLAINS: BMNH_ 2018.2750, BMNH

2018.2748 (Fig. 7F); SIOMA NGWEZI NP: BMNH

2018.2745. Comments: All specimens are juveniles and

were found in miombo woodland. Sequence similarity

is 99% with [. capensis from Katima Mulilo, Namibia

(GenBank accession number JX962898).

Meroles squamulosus (Peters, 1854)

Savanna Lizard

Amphib. Reptile Conserv.

Material. NANZILA PLAINS: BMNH _ 2018.2753

(Fig. 7G); SIOMA NGWEZI NP: BMNH 2018.2752.

Comments: Two adult males found in miombo.

Sequence similarity 1s 96% with Meroles (Ichnotropis)

squamulosus from Laela, Tanzania (GenBank accession

number JX962897).

Scincidae

Panaspis cf. wahlbergi (Smith, 1849)

Snake-eyed Skink

Material. CHAVUMA FR: BMNH 2018.2738, BMNH

2018.2739. Comments: Specimens found during the day

in leaf litter. The closest match from GenBank (98%) is

Panaspis sp. (KU236726), from Katanga, DRC. Medina

et al. (2016) provide a molecular phylogeny of this genus,

which suggests that there is cryptic diversity within P.

wahlbergi.

Trachylepis cf. albopunctata (Bocage, 1867)

Angolan Variable Skink

Material. ITEZHI-TEZHI: BMNH — 2018.2762;

MAYUKUYUKU: BMNH_ 2018.2765 (Fig. 7H),

BMNH_ 2018.2763. Comments: Specimens found

during the day. The BLAST search shows 99-100%

sequence similarity with sequences from 7! varia clade B

(accession numbers MG605651—59), which was recently

assigned to Trachylepis cf. albopunctata by Marques et

al. (2018).

Trachylepis damarana (Peters, 1870)

Damara Skink

Material. SIOMA NGWEZI NP (HQ): BMNH

2018.2764. Comments: Morphologically similar to 7.

varia group. However, both its distribution and sequence

similarity (99%) match T. damarana (see Weinell and

Bauer 2018).

Trachylepis wahlbergii (Peters, 1869)

Wahlberg’s Striped Skink

Material. ITEZHI-TEZHI: BMNH 2018.2769 (Fig.

71); LUKWAKWA: BMNH = 2018.2768, BMNH

2018.2770; NK WAJI: BMNH 2018.2771. Comments:

Specimens found during the day on rocks (Itezhi-Tezhi),

dambo (Lukwakwa), and inside a tree trunk at Nkwaji.

According to Broadley (2000) these specimens fall in

the distribution range of 7. wahlbergii. The most similar

sequence from GenBank is 7’ wahlbergii from Zambia

(99%, accession number DQ234810).

Typhlacontias rohani Angel, 1923

Rohan's Blind Dart Skink

Material. SIOMA NGWEZI NP (HQ): BMNH

2018.2761 (Fig. 7J). Comments: One specimen was

found buried in sand and collected by Errol Pietersen. The

closest match on GenBank (90%) is 7. punctatissimus

(DQ316889). There is no sequence of 7. rohani available

August 2019 | Volume 13 | Number 2 | e181

Herpetological survey of western Zambia

Fig. 8. Drawings of Rhamnophis aethiopissa ituriensis (BMNH

2018.2772). Head and anterior of body in dorsal, left lateral,

and ventral views. Scale bar 10 mm. Drawings by Ed Wade.

for comparison.

Colubridae

Crotaphopeltis hotamboeia (Laurenti, 1768)

White-lipped Herald Snake

Material. ITEZHI-TEZHI: BMNH 2018.2776 (Fig.

7K); HILLWOOD FARM: BMNH 2018.2773; SIOMA

NGWEZI NP: BMNH 2018.2777. Comments: All

individuals collected are juveniles and were found at

night in miombo woodland near rocks. The most similar

sequence on GenBank is C. hotamboeia from Malawi

(99%, accession number AY611816).

Philothamnus hoplogaster (Gunther, 1863)

Green Water Snake

Material. NANZILA PLAINS: BMNH 2018.2775 (Fig.

7L). Comments: Specimen found while eating tadpoles

and juveniles of Kassina sp. in a temporary pond during

the day. This species is similar to other Philothamnus

but usually smaller (Marais 2004). The closest match

on GenBank (99%) is P. hoplogaster from Mozambique

(accession number FJ913484).

Rhamnophis aethiopissa ituriensis (Schmidt 1923)

Large-eyed Green Tree Snake

Material. HILLWOOD FARM: BMNH 2018.2772 (Fig.

7M; Fig. 8). Comments: Specimen found in leaf litter of

riverine forest during the day. Broadley (1991) provided

the first record of this species for Zambia. Based on

distribution, this form represents the subspecies R. a.

ituriensis from Niapu in the DRC (see Eimermacher

2012).

Amphib. Reptile Conserv.

Thelotornis kirtlandii (Hallowell, 1844)

Forest Vine Snake

Material. HILLWOOD FARM: BMNH 2018.2760

(Fig. 7N). Comments: One juvenile was found at night

resting on green vegetation in riverine forest (mushito).

Species was identified using the key in Broadley (2001)

and the following characters were observed: top of

the head, including temporal region, is uniform green;

rostral and nasals are strongly recurved onto top of snout;

supralabials are white with small green spots.

Lamprophiidae

Atractaspis congica Peters, 1877

Congo Stiletto Snake

Material. HILLWOOD FARM: BMNH 2018.2274

(Fig. 70). Comments: One relatively large specimen

found in moist leaf litter inside a patch of mushito at

night. Broadley and Blaylock (2013): 232 expanded the

description of A. bibronii to accommodate the condition

of 19 midbody scales of A. congica (19-21 scales at

midbody). This specimen exhibits erratic counts of

19+17, the latter count predominating after the 84"

ventral.

Order Testudines

Testudinidae

Kinixys spekii Gray, 1863

Speke’s Hinge-back Tortoise

Comments: One specimen (Figs. 5P—R) was found in

mushito at Nkwaji. The carapace was hinged between the

7" and 8" marginals. The specimen was photographed

and released.

Discussion

This is a non-comprehensive list of the herpetofauna of

western Zambia. The survey was conducted shortly after

the general breeding season for amphibians and reptiles in

this part of the world, and consequently most specimens

collected are juveniles. For the same reason, most species

were not active, which made the search for them more

challenging. However, some species of amphibians were

active during the survey. Phrynobatrachus were heard

calling during the day and at night, and at least ten males

of Sclerophrys lemairii were calling during the day in

Lukwakwa (see Bittencourt-Silva 2014). The presence of

juveniles of /chnotropis capensis and adults of Meroles

squamulosus in sympatry is explained by their staggered

life cycles (see Broadley 1967, 1979).

DNA barcoding is an important tool for identifying

candidate species. However, there are a number of

caveats. For instance, for amphibians, Vences et al.

(2005a) propose a tentative 16S rRNA threshold at

5% for interspecific sequence divergence but also

August 2019 | Volume 13 | Number 2 | e181

Bittencourt-Silva

highlight the broad overlap of intra- and interspecific

divergence values (see Vences et al. 2005b, p. 1,865)

that complicates the establishment of threshold values.

Using DNA barcoding alone can potentially lead to

simplistic diagnoses of putative species. Another issue

with DNA barcoding relates to the taxonomic accuracy

of public DNA databases (e.g., GenBank, BOLD,

EMBL). Misidentified sequences are not uncommon

(Bridge et al. 2003; Vilgalys 2003), which reinforces

the importance of vouchering all sequences deposited.

Table 2 shows that the intra-specific p-distances within

H. marmoratus are considerably large in some cases.

This could be partly due to geographic distances, given

that the specimens are from Ghana, Guinea-Bissau,

Kenya, and Tanzania. A taxonomic review of this group

is clearly necessary. Nonetheless, genetic data may be

crucial in cases where species are genetically different

but morphologically largely conserved. An example is

the mongrel frogs from Mozambique and Malawi, which

have ca. 5% interspecific divergences but are in general

phenotypically indistinguishable (Conradie et al. 2018).

Some of the taxa reported here could not be assigned

to currently recognized species based on DNA barcoding

and/or external morphology. For instance, based on

p-distances, some of the Phrynobatrachus specimens

represent putative new species (Table 3). Zimkus and

Schick (2010) suggest that there are at least two species

currently identified as Phrynobatrachus natalensis in

East Africa, and another two clades are reported from

western and southern Africa (Zimkus et al. 2010). The

phylogeny presented here indicates that there may be

more species of this group in western Zambia (see Fig.

5). Species identified here as P. cf. parvulus may be a

new species. These results corroborate the conclusions

of Zimkus and Schick (2010) and Zimkus et al. (2010)

that a taxonomic review of the genus Phrynobatrachus

is needed. Similarly, the Zomopterna population found in

Sioma Ngwezi NP could represent an undescribed species

(Table 4) previously reported from Namibia (see Dawood

et al. 2002). These taxa deserve further investigations

by specialists. It is often the case that original species

descriptions lack diagnostic details, including illustration

of characters, and/or type material may be lost or in poor

condition, all of which can contribute to inconclusive or

even incorrect species identification. Additional datasets

(e.g., bioacoustics, ecology) often provide important

information and can solve some of these taxonomic

conundrums.

The genus Ptychadena currently comprises 56

species, some (possibly many) representing species

complexes (e.g., Zimkus et al. 2016). Nineteen species

have been reported in Zambia (see genus account in

Frost 2018), eleven of which were recorded during this

survey. The lack of an updated key to the Ptychadena

of Zambia makes the species identification process

challenging. Similarly, barcoding is not helpful when

there are no reference sequences available. A search of

Amphib. Reptile Conserv.

GenBank for 16S sequences of Ptychadena shows that

41% of currently recognized species are not represented,

and 22% of the sequences available are either pending

confirmation or identified only to genus. The taxonomy

of this group is clearly in need of attention.

Except for a few areas—on the extreme north-west of

the country and along the Zambezi river—most of Zambia

remains poorly studied. Recently, Channing and Willems

(2018) described a new species of Ptychadena from the

northern part of the country, and a new cryptic species of

Polemon (Squamata: Lamprophiidae) described from the

DRC and Uganda is likely to occur in Zambia (Portillo

et al. 2019). The list of species provided here adds new

points to the map of the Zambian herpetofauna.

The herpetofauna of Zambia is mostly contained

in the Zambezian biogeographical core, with only the

south-western region forming part of the South African

core (sensu Linder et al. 2012). Not surprisingly, many

species found during this survey also occur in the DRC

(e.g., Channing et al. 2016), Angola (Conradie et al. 2016)

and Namibia (Dawood et al. 2002), including Amietia

chapini, recorded here for the first time from Zambia.

Four species of amphibians that Pietersen et al. (2017)

expected to occur near Ngonye Falls are now confirmed

to occur at Sioma Ngwezi NP (Kassina senegalensis,

Phrynobatrachus mababiensis, Ptychadena porosissima,

and Pyxicephalus adspersus). The still very incomplete

knowledge of the Zambian herpetofauna remains the

main obstacle to our understanding of its biogeography

and the conservation statuses of its constituent species.

Acknowledgements.—| thank the entomologists Hitoshi

Takano, Lucia Chmurova, and Lydia Smith for support in

the field. I am grateful to Errol Pietersen for assisting with

logistics and collection of specimens at Ngonye Falls,

and to Darren Pietersen for helping with identification of

some reptiles and providing valuable comments on the

manuscript. The expedition to Zambia would not have

been possible without the support of Richard Smith, to

whom I am extremely grateful. I thank Werner Conradie

and Mark Wilkinson for their valuable comments on the

manuscript. Ed Wade kindly provided the scale counting

of Atractaspis congica and the drawings of Rhamnophis

aethiopissa ituriensis. | thank Simon P. Loader for his

invaluable support. I appreciate the help of Dan Portik

and Breda Zimkus with identification of Hyperolius and

Phrynobatrachus, respectively. Permits to collect and

export specimens were issued by the Department of

Veterinary Services, Ministry of Livestock and Fisheries

Development (ICS#08417).

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Gabriela Bittencourt is a Brazilian evolutionary biologist with research experience in natural

history, evolution, ecology, and biogeography, and a particular focus on amphibians. Gabriela

has more than 15 years of herpetological laboratory and fieldwork experience in the Neo-

tropics, Africa, and Asia. Her research has focused on understanding phylogenetic relation-

ships and biotic distribution patterns of amphibians. Gabriela is currently a Research Assistant

in the Herpetology Group at the Natural History Museum, London, United Kingdom.

August 2019 | Volume 13 | Number 2 | e181

Bittencourt-Silva

Appendix 1. List of amphibians and reptiles found in western Zambia, including species vouchers, GenBank accession numbers, and

locality information. Museum acronym: BMNH — Natural History Museum, London, United Kingdom. GPS datum WGS-84.

Species

AMPHIBIA: ANURA

ARTHROLEPTIDAE

Arthroleptis stenodactylus

Arthroleptis xenochirus

BUFONIDAE

Sclerophrys gutturalis

Sclerophrys lemairii

Sclerophrys pusilla

Voucher ID

BMNH 2018.5826

BMNH 2018.5827

BMNH 2018.5828

BMNH 2018.5829

BMNH 2018.5830

BMNH 2018.5814

BMNH 2018.5815

BMNH 2018.5816

BMNH 2018.5817

BMNH 2018.5818

BMNH 2018.5819

BMNH 2018.5820

BMNH 2018.5811

BMNH 2018.5812

BMNH 2018.5813

BMNH 2018.5821

BMNH 2018.5822

BMNH 2018.5823

BMNH 2018.5824

BMNH 2018.5825

BMNH 2018.5703

BMNH 2018.5705

BMNH 2018.5706

BMNH 2018.5707

BMNH 2018.5702

BMNH 2018.5701

BMNH 2018.5704

BMNH 2018.5723

BMNH 2018.5715

BMNH 2018.5716

BMNH 2018.5717

BMNH 2018.5718

BMNH 2018.5719

BMNH 2018.5720

BMNH 2018.5721

BMNH 2018.5722

BMNH 2018.5709

BMNH 2018.5710

BMNH 2018.5708

BMNH 2018.5711

Amphib. Reptile Conserv.

Field ID GenBank Locality Latitude

SL2109 MK464479 ~~ Lukwakwa -12.66084

SL2121 | MK464478 ~~ Lukwakwa -12.66084

SL2123, MK464477_ ~— Lukwakwa -12.66084

SL2128 = MK464476 ~~ Lukwakwa -12.66084

SL2221 MK464475 — Nkwaji -11.57728

SL2145 =MK464471 — Hillwood Farm -11.26316

SL2146 =MK464470 — Hillwood Farm -11.26316

SL2147, MK464469 — Hillwood Farm -11.26316

SL2148 = MK464468 — Hillwood Farm -11.26316

SL2152. MK464467 — Hillwood Farm -11.26690

SL2153. MK464466 — Hillwood Farm -11.26690

SL 2246 MK464465 — Hillwood Farm -11.26690

SL2122 MK464474 ~~ Lukwakwa -12.66084

SL2124 MK464473 ~~ Lukwakwa -12.66084

SL2125 MK464472 ~~ Lukwakwa -12.66084

SL 2200 MK464464 Nkwaji -11.60592

SL 2201 MK464463 = Nkwaji -11.60592

SL 2202 MK464462 = Nkwaji -11.60592

SL 2203. MK464461 Nkwaji -11.60592

SL2204 MK464460 = Nkwaji -11.60592

SL2102 Chavuma FR -13.07006

SL2108 = MK464294 ~~ Lukwakwa -12.66084

SL2107 MK464293 ~Lukwakwa -12.74275

SL 2111 Lukwakwa -12.66084

SL 2069 MK464296 Maramba Lodge, Livingstone -17.89120

SL2025 MK464297 = Mayukuyuku, Kafue NP -14.91533

SL2190 MK464295 Nkwaji -11.60592

SL 2245 Hillwood Farm -11.26690

SL 2112 Lukwakwa -12.66084

SL2113 Lukwakwa -12.66084

SL 2114 Lukwakwa -12.66084

SL 2115 Lukwakwa -12.66084

SL 2116 Lukwakwa -12.66084

SL 2117 Lukwakwa -12.66084

SL 2118 Lukwakwa -12.66084

SL 2119 Lukwakwa -12.66084

SL 2047. MK464291 _ Itezhi-Tezhi, Kafue NP -15.77340

SL 2053. MK464290 _ Itezhi-Tezhi, Kafue NP -15.77340

SL2012 MK464292. Mayukuyuku, Kafue NP -14.91533

SL2189 MK464289 = Nkwaji -11.60592

Longitude

24.43697

24.53960

24.32782

24.31666

24.43697

24.55448

22.92880

24.43697

24.28436

24.43697

25.85821

26.06311

24.55448

24.31666

24.43697

26.01151

26.06311

24.55448

Altitude

1063

1291

1356

1308

1063

1244

1073

1063

1101

1063

900

1012

1244

1308

1063

1036

1012

1244

August 2019 | Volume 13 | Number 2 | e181

Herpetological survey of western Zambia

Appendix 1 (continued). List of amphibians and reptiles found in western Zambia, including species vouchers, GenBank accession

numbers, and locality information. Museum acronym: BMNH — Natural History Museum, London, United Kingdom. GPS datum WGS-84.

Species Voucher ID Field ID GenBank Locality Latitude Longitude Altitude

Sclerophrys pusilla BMNH 2018.5712 SL2191 MK464288 = Nkwaji -11.60592 24.55448 1244

Schismaderma carens BMNH 2018.5729 SL2105 MK464298 Chavuma FR -13.07006 22.92880 1073

Schismaderma carens BMNH 2018.5724 SL2048 MK464303 _ Itezhi-Tezhi, Kafue NP -15.77340 26.01151 1036

Schismaderma carens BMNH 2018.5725 SL2049 MK464302 _ Itezhi-Tezhi, Kafue NP -15.77340 26.01151 1036

Schismaderma carens BMNH 2018.5726 SL2050 MK464301 _ Itezhi-Tezhi, Kafue NP -15.77340 26.01151 1036

Schismaderma carens BMNH 2018.5727 =SL 2051 MK464300 _ Itezhi-Tezhi, Kafue NP -15.77340 26.01151 1036

Schismaderma carens BMNH 2018.5728 SL2052 MK464299 _ Itezhi-Tezhi, Kafue NP -15.77340 26.01151 1036

HEMISOTIDAE

Hemisus cf. guineensis BMNH 2018.5801 SL2252 MK464449 — Hillwood Farm -11.26690 24.31666 1308

Hemisus cf. guineensis BMNH 2018.5800 SL2110 MK464450 Lukwakwa -12.66084 24.43697 1063

Hemisus cf. guineensis BMNH 2018.5799 SL 2091 MK464451 Sioma Ngwezi NP -16.89873 23.59847 1009

Hemisus marmoratus BMNH 2018.5713 SL2018 = MK464448 Mayukuyuku, Kafue NP -14.91533 26.06311 1012

Hemisus marmoratus BMNH 2018.5714 SL2090 MK464447 Sioma Ngwezi NP -16.89873 23.59847 1009

HYPEROLIIDAE

Hyperolius dartevellei BMNH 2018.5681 SL2098 MK464446 Chavuma FR -13.07006 22.92880 1073

Hyperolius dartevellei BMNH 2018.5683 SL2139 MK464444 ~— Hillwood Farm -11.26690 24.31666 1308

Hyperolius dartevellei BMNH 2018.5684 SL2140 MK464443 ~~ Hillwood Farm -11.26690 24.31666 1308

Hyperolius dartevellei BMNH 2018.5682, SL2127 MkK464445 ~~ Lukwakwa -12.66084 24.43697 1063

Hyperolius kachalolae BMNH 2018.5676 SL2138 MK464442 ~~ Hillwood Farm -11.26690 24.31666 1308

Hyperolius kachalolae BMNH 2018.5677 = SL2149. MkK464441 ~— Hillwood Farm -11.26316 24.32782 1356

Hyperolius kachalolae BMNH 2018.5678 SL2180 MK464440 — Hillwood Farm -11.26690 24.31666 1308

Hyperolius kachalolae BMNH 2018.5679 SL2243 MK464439 — Hillwood Farm -11.26690 24.31666 1308

Hyperolius kachalolae BMNH 2018.5680 SL2244 MK464438 — Hillwood Farm -11.26690 24.31666 1308

Hyperolius major BMNH 2018.5675 SL2159 MK464437 ~— Hillwood Farm -11.27444 24.32444 1416

Hyperolius marginatus BMNH 2018.5667 SL2126 MK464436 Lukwakwa -12.66091 24.42943 1100

Hyperolius marginatus BMNH 2018.5668 SL2063 MK464435 ~ Nanzila Plains, Kafue NP -16.28138 25.91676 L032

Hyperolius marginatus BMNH 2018.5674 SL2064 MK464434 = Nanzila Plains, Kafue NP -16.28138 25.91676 1032

Hyperolius nasicus BMNH 2018.5666 SL2056 MK464433 ~ Nanzila Plains, Kafue NP -16.28138 25.91676 1032

Hyperolius paralellus BMNH 2018.5687) = SL2133) MK464432 ~— Hillwood Farm -11.26690 24.31666 1308

Hyperolius paralellus BMNH 2018.5688 SL2136 MK464431 Hillwood Farm -11.26690 24.31666 1308

Hyperolius paralellus BMNH 2018.5689 SL2137 MK464430 — Hillwood Farm -11.26690 24.31666 1308

Hyperolius paralellus BMNH 2018.5690 SL 2141 MK464429 — Hillwood Farm -11.26690 24.31666 1308

Hyperolius paralellus BMNH 2018.5691 SL2142 MK464428 — Hillwood Farm -11.26690 24.31666 1308

Hyperolius paralellus BMNH 2018.5692, SL2150 MK464427 ~~ Hillwood Farm -11.26316 24.32782 1356

Hyperolius paralellus BMNH 2018.5693 SL2161 MK464426 ~— Hillwood Farm -11.27444 24.32444 1416

Hyperolius paralellus BMNH 2018.5694 SL2179 MK464425 — Hillwood Farm -11.26690 24.31666 1308

Hyperolius paralellus BMNH 2018.5695 SL2184 Hillwood Farm -11.26690 24.31666 1308

Hyperolius paralellus BMNH 2018.5696 SL2224 MK464424 = Nkwaji -11.50420 24.56456 1386

Hyperolius paralellus BMNH 2018.5697 = SL2225 MK464423 Nkwaji -11.50420 24.56456 1386

Hyperolius paralellus BMNH 2018.5698 SL2220 MK464422 Nkwaji -11.60592 24.55448 1244

Hyperolius paralellus BMNH 2018.5699 SL2240 MK464421 Nkwaji -11.53906 24.55262 1336

Hyperolius paralellus BMNH 2018.5700 SL 2241 MK464420 Nkwaji -11.53906 24.55262 1336

Hyperolius quinquevittatus BMNH 20185685 SL2216 MK464419 = Nkwaji -11.53906 24.55262 1336

Amphib. Reptile Conserv. 22 August 2019 | Volume 13 | Number 2 | e181

Bittencourt-Silva

Appendix 1 (continued). List of amphibians and reptiles found in western Zambia, including species vouchers, GenBank accession

numbers, and locality information. Museum acronym: BMNH — Natural History Museum, London, United Kingdom. GPS datum WGS-84.

Species Voucher ID Field ID GenBank Locality Latitude Longitude Altitude

Hyperolius quinquevittatus BMNH 2018.5686 SL 2217 Nkwaji -11.53906 24.55262 1336

Kassina senegalensis BMNH 2018.5810 SL2106 | MK464406 Chavuma FR -13.07006 22.92880 1073

Kassina senegalensis BMNH 2018.5802 SL2129 MK464414 Hillwood Farm -11.26690 _24,31666 1308

Kassina senegalensis BMNH 2018.5803. SL2130 MK464413 Hillwood Farm -11.26690 —_24,31666 1308

Kassina senegalensis BMNH 2018.5804. SL2223. MK464412 — Nkwaji -11.50420 24,56456 1386

Kassina senegalensis BMNH 2018.5805 SL2073 MK464411 — Sioma Ngwezi NP -16.89873 23.59847 1009

Kassina senegalensis BMNH 2018.5806 SL2074 MK464410 — Sioma Ngwezi NP -16.89873 23.59847 1009

Kassina senegalensis BMNH 2018.5807 SL2075 MK464409 — Sioma Ngwezi NP -16.89873 2359847 1009

Kassina senegalensis BMNH 2018.5808 SL2076 MK464408 — Sioma Ngwezi NP -16.89873 23.59847 1009

Kassina senegalensis BMNH 2018.5809 SL2087 MK464407_ — Sioma Ngwezi NP -16.89873 23.59847 1009

PHRYNOBATRACHIDAE

| ee ee ee BMNH 2018.5873. SL2157 | MK464397 Hillwood Farm -11.26316 24,32782 1356

i‘ eet, of BMNH 2018.5874. SL2162 MK464396 Hillwood Farm -11.27444 2432444 1416

‘ per ade cL BMNH 2018.5875 SL2163 MK464395 Hillwood Farm -11.26316 24.32782 1356

; ieee akin i eae BMNH 2018.5876 SL2164 | MK464394 Hillwood Farm -11.26316 24,32782 1356

ee o BMNH 2018.5877. SL2165 = _MK464393 Hillwood Farm -11.26316 24,32782 1356

i ited dar - BMNH 2018.5878 SL2166 MK464392 Hillwood Farm -11.26316 24.32782 1356

; ecaireteabe 7 BMNH 2018.5889 SL2120 MK464389 — Lukwakwa -12.66084 24.43697 1063

Phrynobatrachus cf. . oft sh

BMNH 2018.5879 SL2208 MK464391 = Nkwaji -11.56594. —.24.52659 1311

parvulus

Phrynobatrachus cf. : PA dae

BMNH 2018.5880 SL2210 MK464390 — Nkwaji -11.56594 —.24,52659 1311

parvulus

PRODI CE CE BMNH 2018.5882 SL 2205 Nkwaji -11.56594. 24.52659 1311

parvulus

DL yO RGIrabiRE Cl: BMNH 2018.5883 SL 2234 Nkwaji -11.56567 — 24.52605 1263

parvulus

ER vier eiey deus Ch BMNH 2018.5884 SL 2235 Nkwaji -11.56567 — 24.52605 1263

parvulus

RD ACHES, BMNH 2018.5885 SL 2236 Nkwaji -11.56567 —_24.52605 1263

parvulus

MEADE CRUSE: BMNH 2018.5886 SL 2237 Nkwaji -11.56567 — 24.52605 1263

parvulus

Hey CRO aChHis- eh. BMNH 2018.5887 SL 2238 Nkwaji -11.56567 — 24.52605 1263

parvulus

PIN RCDGI OCHS Che BMNH 2018.5888 SL 2239 Nkwaji -11.56567 — 24.52605 1263

parvulus

Pu ODER ACEUS BMNH 2018.5831 SL2010 MK464388 = Mayukuyuku, Kafue NP -14.91533 26.06311 1012

mababiensis

BN OHA, Aas BMNH 2018.5832. SL2011 = MK464387 = Mayukuyuku, Kafue NP -14.91533 26.0631 1012

mababiensis

Phrynobatrachus Es a

oe BMNH 2018.5881 SL 2015 Mayukuyuku, Kafue NP -14.91533 26.06311 1012

mababiensis

Ripry robin achis BMNH 2018.5833 SL2065 MK464386 = Nanzila Plains, Kafue NP -16.28138 25.91676 1032

mababiensis

BP levnoparr actus BMNH 2018.5834. SL2080 MK464385 — Sioma Ngwezi NP -16.89873 23.59847 1009

mababiensis

Amphib. Reptile Conserv. 23 August 2019 | Volume 13 | Number 2 | e181

Herpetological survey of western Zambia

Appendix 1 (continued). List of amphibians and reptiles found in western Zambia, including species vouchers, GenBank accession

numbers, and locality information. Museum acronym: BMNH — Natural History Museum, London, United Kingdom. GPS datum WGS-84.

Species Voucher ID Field ID GenBank Locality Latitude Longitude Altitude

pI BOE ET aCiES. BMNH 2018.5835 SL 2081 Sioma Ngwezi NP -16.89873 23.59847 1009

mababiensis

Phrynobatrachus natalensis BMNH 2018.5848 SL2045 MK464377 _ Itezhi-Tezhi, Kafue NP -15.77340 26.01151 1036

Phrynobatrachus natalensis BMNH 2018.5849 SL2046 MK464376 _ Itezhi-Tezhi, Kafue NP -15.77340 26.0115] 1036

Phrynobatrachus natalensis +BMNH 2018.5850 SL2054 MK464375 _ Itezhi-Tezhi, Kafue NP -15.77340 26.01151 1036

Phrynobatrachus natalensis BMNH 2018.5853 SL2066 = MK464372 Maramba Lodge, Livingstone -17.89120 25.8582] 900

Phrynobatrachus natalensis BMNH 2018.5854 SL2067 MK464371 Maramba Lodge, Livingstone -17.89120 25.8582] 900

Phrynobatrachus natalensis BMNH 2018.5837. SL2016 MK464384 Mayukuyuku, Kafue NP -14.91533 26.06311 1012

Phrynobatrachus natalensis BMNH 2018.5838 SL2017 = MK464383 Mayukuyuku, Kafue NP -14.91533 26.06311 1012

Phrynobatrachus natalensis BMNH 2018.5839 SL2026 MK464382 Mayukuyuku, Kafue NP -14.91533 26.06311 1012

Phrynobatrachus natalensis BMNH 2018.5840 SL2027 MK464381 Mayukuyuku, Kafue NP -14.91533 26.06311 1012

Phrynobatrachus natalensis BMNH 2018.5841 SL2033 MK464380 Mayukuyuku, Kafue NP -14.91533 26.06311 1012

Phrynobatrachus natalensis BMNH 2018.5842 SL2034 = MK464379 Mayukuyuku, Kafue NP -14.91533 26.06311 1012

Phrynobatrachus natalensis BMNH 2018.5843 SL2037 MK464378 Mayukuyuku, Kafue NP -14.91533 26.06311 1012

Phrynobatrachus natalensis BMNH 2018.5844 SL 2028 Mayukuyuku, Kafue NP -14.91533 26.06311 1012

Phrynobatrachus natalensis BMNH 2018.5845 = SL 2029 Mayukuyuku, Kafue NP -14.91533 26.06311 1012

Phrynobatrachus natalensis BMNH 2018.5846 — SL 2030 Mayukuyuku, Kafue NP -14.91533 26.06311 1012

Phrynobatrachus natalensis BMNH 2018.5847 = SL 2031 Mayukuyuku, Kafue NP -14.91533 26.06311 1012

Phrynobatrachus natalensis BMNH 2018.5851 SL2055 MK464374 Nanzila Plains, Kafue NP -16.28138 25.91676 1032

Phrynobatrachus natalensis BMNH 2018.5852. SL2057 MkK464373 ~~ Nanzila Plains, Kafue NP -16.28138 25.91676 1032

Phrynobatrachus sp.1 BMNH 2018.5866 SL2143 MK464364 ~— Hillwood Farm -11.26316 24.32782 1356

Phrynobatrachus sp.1 BMNH 2018.5867 SL2144 MK464363 ~~ Hillwood Farm -11.26316 24.32782 1356

Phrynobatrachus sp.1 BMNH 2018.5856 SL2195 MK464369 = Nkwaji -11.60592 24.55448 1244

Phrynobatrachus sp.1 BMNH 2018.5857. SL 2196 Nkwaji -11.60592 24.55448 1244

Phrynobatrachus sp.1 BMNH 2018.5858 SL2197 MK464368 Nkwaji -11.60592 24.55448 1244

Phrynobatrachus sp.1 BMNH 2018.5859 SL2198 = MK464367 = Nkwaji -11.60592 24.55448 1244

Phrynobatrachus sp.1 BMNH 2018.5860 SL2199 MK464366 Nkwaji -11.60592 24.55448 1244

Phrynobatrachus sp.1 BMNH 2018.5861 SL 2211 Nkwaji -11.56594 24.52659 1311

Phrynobatrachus sp.1 BMNH 2018.5862 SL 2219 Nkwaji -11.56567 24.52605 1263

Phrynobatrachus sp.1 BMNH 2018.5863 SL2232 MK464365 Nkwaji -11.56567 24.52605 1263

Phrynobatrachus sp.1 BMNH 2018.5868 SL2206 MK464362 Nkwaji -11.56594 24.52659 131]

Phrynobatrachus sp.1 BMNH 2018.5869 SL2207 MK464361 Nkwaji -11.56594 24.52659 1311

Phrynobatrachus sp.1 BMNH 2018.5870 SL2209 MK464360 Nkwaji -11.56594 24.52659 jevel

Phrynobatrachus sp.1 BMNH 2018.5871 SL2231 MK464359 = Nkwaji -11.56567 24.52605 1263

Phrynobatrachus sp.1 BMNH 2018.5872, SL2233) MK464358 Nkwaji -11.56567 24.52605 1263

Phrynobatrachus sp.2 BMNH 2018.5836 SL2071 #MK464357 Ngonye Falls Camp -16.66139 23.57280 929

Phrynobatrachus sp.2 BMNH 2018.5864 SL2095 MK464356 Sioma Ngwezi NP -16.89873 23.59847 1009

Phrynobatrachus sp.2 BMNH 2018.5865 SL2096 MK464355 Sioma Ngwezi NP -16.89873 23.59847 1009

Phrynobatrachus sp.3 BMNH 2018.5855 SL2072 MK464370 Ngonye Falls Camp -16.66139 23.57280 929

PIPIDAE

Xenopus poweri BMNH 2018.5654 SL2173 MK464274 ~— Hillwood Farm -11.26690 24.31666 1308

Xenopus poweri BMNH 2018.5655 SL2174 =MK464273 ~— Hillwood Farm -11.26690 24.31666 1308

Xenopus poweri BMNH 2018.5656 SL2175 =MK464272 ~— Hillwood Farm -11.26690 24.31666 1308

Amphib. Reptile Conserv. 24 August 2019 | Volume 13 | Number 2 | e181

Bittencourt-Silva

Appendix 1 (continued). List of amphibians and reptiles found in western Zambia, including species vouchers, GenBank accession

numbers, and locality information. Museum acronym: BMNH — Natural History Museum, London, United Kingdom. GPS datum WGS-84.

Species Voucher ID Field ID GenBank Locality Latitude Longitude Altitude

Xenopus poweri BMNH 2018.5657, = SL2176 MK464271 ~— Hillwood Farm -11.26690 24.31666 1308

Xenopus poweri BMNH 2018.5658 SL 2177 Hillwood Farm -11.26690 24.31666 1308

Xenopus poweri BMNH 2018.5659 SL2222 MK464270 = Nkwaji -11.50420 24.56456 1386

Xenopus pygmaeus BMNH 2018.5651 SL2134 MK464269 — Hillwood Farm -11.26690 24.31666 1308

Xenopus pygmaeus BMNH 2018.5652, SL2135 MK464268 — Hillwood Farm -11.26690 24.31666 1308

Xenopus pygmaeus BMNH 2018.5653 SL2156 MK464267 ~— Hillwood Farm -11.26690 24.31666 1308

PTYCHADENIDAE

Ptychadena anchietae BMNH 2018.5735 SL2040 MK464344 _ Itezhi-Tezhi, Kafue NP -15.7734 26.01151 1036

Ptychadena anchietae BMNH 2018.5736 SL 2039 Itezhi-Tezhi, Kafue NP -15.7734 26.01151 1036

Ptychadena anchietae BMNH 2018.5730 SL 2019 Mayukuyuku, Kafue NP -14.91533 26.06311 1012

Ptychadena anchietae BMNH 2018.5731 SL2020 MK464345 Mayukuyuku, Kafue NP -14.91533 26.06311 1012

Ptychadena anchietae BMNH 2018.5732 SL 2022 Mayukuyuku, Kafue NP -14.91533 26.06311 1012

Ptychadena anchietae BMNH 2018.5733 SL 2023 Mayukuyuku, Kafue NP -14.91533 26.06311 1012

Ptychadena anchietae BMNH 2018.5734 SL 2024 Mayukuyuku, Kafue NP -14.91533 26.06311 1012

Ptychadena grandisonae BMNH 2018.5737. = SL2214 MK464318 Nkwaji -11.5042 24.56456 1386

Ptychadena grandisonae BMNH 2018.5738 SL 2215 Nkwaji -11.5042 24.56456 1386

Ptychadena grandisonae BMNH 2018.5739 SL2226 MK464317 = Nkwaji -11.5042 24.56456 1386

Ptychadena grandisonae BMNH 2018.5740 SL2227 MK464316 Nkwaji -11.5042 24.56456 1386

Ptychadena grandisonae BMNH 2018.5741 SL2228 MK464315 = Nkwaji -11.5042 24.56456 1386

Ptychadena grandisonae BMNH 2018.5742 SL2229 MK464314 = Nkwaji -11.5042 24.56456 1386

Ptychadena grandisonae BMNH 2018.5743, SL2230 MK464313 Nkwaji -11.5042 24.56456 1386

Ptychadena cf. guibei BMNH 2018.5764 SL2035 MK464324 Mayukuyuku, Kafue NP -14.91533 26.06311 1012

Ptychadena cf. guibei BMNH 2018.5765 SL2079 MK464323 Sioma Ngwezi NP -16.89873 23.59847 1009

Ptychadena mapacha BMNH 2018.5772 SL2014 MK464312 Mayukuyuku, Kafue NP -14.91533 26.06311 1012

Ptychadena cf. mossambica BMNH 2018.5754 SL2042 MK464342 _ Itezhi-Tezhi, Kafue NP -15.7734 26.01151 1036

Ptychadena cf. mossambica BMNH 2018.5755 = SL2043. MK464341 _ Itezhi-Tezhi, Kafue NP -15.7734 26.01151 1036

Ptychadena cf. mossambica BMNH 2018.5756 SL2044 MK464340 — Itezhi-Tezhi, Kafue NP -15.7734 26.01151 1036

Ptychadena cf. mossambica BMNH 2018.5757. =SL2038 MK464339 Itezhi-Tezhi, Kafue NP -15.7734 26.01151 1036

Ptychadena cf. mossambica =BMNH 2018.5763 = SL 2021 MK464336 = Mayukuyuku, Kafue NP -14.91533 26.06311 1012

Ptychadena cf. mossambica BMNH 2018.5758 SL 2088 Sioma Ngwezi NP -16.89873 23.59847 1009

Ptychadena cf. mossambica BMNH 2018.5759 SL2089 MK464338 Sioma Ngwezi NP -16.89873 23.59847 1009

Ptychadena cf. mossambica + BMNH 2018.5760 SL2094 MK464337 Sioma Ngwezi NP -16.89873 23.59847 1009

Ptychadena cf. mossambica ~=BMNH 2018.5761 Sioma Ngwezi NP -16.89873 23.59847 1009

Ptychadena cf. mossambica BMNH 2018.5753. SL 2041 Itezhi-Tezhi, Kafue NP -15.7734 26.01151 1036

Ptychadena nilotica BMNH 2018.5781 SL 2068 =MK464327 Maramba Lodge, Livingstone -17.8912 25.85821 900

Ptychadena nilotica BMNH 2018.5773 = SL2058 MK464332 = Nanzila Plains, Kafue NP -16.28138 25.91676 1032

Ptychadena nilotica BMNH 2018.5774 SL2059 MK464331 Nanzila Plains, Kafue NP -16.28138 25.91676 1032

Ptychadena nilotica BMNH 2018.5775 SL2061 MK464330 ~ Nanzila Plains, Kafue NP -16.28138 25.91676 1032

Ptychadena nilotica BMNH 2018.5776 SL 2062 Nanzila Plains, Kafue NP -16.28138 25.91676 1032

Ptychadena nilotica BMNH 2018.5777, =SL2278 MK464329 Nanzila Plains, Kafue NP -16.28138 25.91676 1032

Ptychadena nilotica BMNH 2018.5778 = SL 2279 Nanzila Plains, Kafue NP -16.28138 25.91676 1032

Ptychadena nilotica BMNH 2018.5779 = SL2280 MK464328 Nanzila Plains, Kafue NP -16.28138 25.91676 1032

Ptychadena nilotica BMNH 2018.5780 SL 2281 Nanzila Plains, Kafue NP -16.28138 25.91676 1032

Amphib. Reptile Conserv. 25 August 2019 | Volume 13 | Number 2 | e181

Herpetological survey of western Zambia

Appendix 1 (continued). List of amphibians and reptiles found in western Zambia, including species vouchers, GenBank accession

numbers, and locality information. Museum acronym: BMNH — Natural History Museum, London, United Kingdom. GPS datum WGS-84.

Species Voucher ID Field ID GenBank Locality Latitude Longitude Altitude

Ptychadena obscura BMNH 2018.5768 SL2155 MK464322 ~— Hillwood Farm -11.2669 24.31666 1308

Ptychadena obscura BMNH 2018.5766 SL2131 MK464311 = Hillwood Farm -11.2669 24.31666 1308

Ptychadena obscura BMNH 2018.5767 = SL2132 MK464310 — Hillwood Farm -11.2669 24.31666 1308

Ptychadena oxyrhynchus BMNH 2018.5783 SL2250 #MK464325 — Hillwood Farm -11.2669 24.31666 1308

Ptychadena oxyrhynchus BMNH 2018.5782. SL2060 MK464326 = Nanzila Plains, Kafue NP -16.28138 25.91676 1032

Ptychadena porosissima BMNH 2018.5769 SL2158 = MK464321 — Hillwood Farm -11.27531 24.31977 1340

Ptychadena porosissima BMNH 2018.5770 SL2168 = MK464320 — Hillwood Farm -11.2669 24.31666 1308

Ptychadena porosissima BMNH 2018.5771 SL2248 = MK464319 — Hillwood Farm -11.2669 24.31666 1308

Ptychadena cf. taenioscelis |» BMNH 2018.5784 SL2169 MK464335 — Hillwood Farm -11.2669 24.31666 1308

Ptychadena cf. taenioscelis = BMNH 2018.5785 SL 2171 MK464334 — Hillwood Farm -11.2669 24.31666 1308

Ptychadena cf. taenioscelis © BMNH 2018.5786 SL2172 MK464333 — Hillwood Farm -11.2669 24.31666 1308

Ptychadena upembae BMNH 2018.5750 SL2154. MK464348 — Hillwood Farm -11.2669 24.31666 1308

Ptychadena upembae BMNH 2018.5751 SL2170 MK464347 ~— Hillwood Farm -11.2669 24.31666 1308

Ptychadena upembae BMNH 2018.5752. SL 2251 MK464346 — Hillwood Farm -11.2669 24.31666 1308

Ptychadena upembae BMNH 2018.5744 SL2192 MK464354 Nkwaji -11.60592 24.55448 1244

Ptychadena upembae BMNH 2018.5745 SL2193 MK464353 Nkwaji -11.60592 24.55448 1244

Ptychadena upembae BMNH 2018.5746 SL2194 MK464352 Nkwaji -11.60592 24.55448 1244

Ptychadena upembae BMNH 2018.5747 SL2212 MK464351 Nkwaji -11.56594 24.52659 1311

Ptychadena upembae BMNH 2018.5748 SL2218 MK464350 Nkwaji -11.53906 24.55262 1336

PyxICEPHALIDAE

Amietia chapini BMNH 2018.5660 SL2186 MK464482 ~~ Hillwood Farm -11.26690 24.31666 1308

Amietia chapini BMNH 2018.5661 SL2185 MK464481 — Hillwood Farm -11.26690 24.31666 1308

Amietia chapini BMNH 2018.5662 SL2178 Hillwood Farm -11.26690 24.31666 1308

Amietia chapini BMNH 2018.5663 SL2183 MK464480 — Hillwood Farm -11.26690 24.31666 1308

Amietia chapini BMNH 2018.5664 SL2151 Hillwood Farm -11.26690 24.31666 1308

Amietia chapini BMNH 2018.5665 SL 2167 Hillwood Farm -11.26690 24.31666 1308

Pyxicephalus cf. adspersus BMNH 2018.5787 SL2077 MK464309 Sioma Ngwezi NP -16.89873 23.59847 1009

Pyxicephalus cf. adspersus BMNH 2018.5788 SL2082 MK464308 Sioma Ngwezi NP -16.89873 23.59847 1009

Pyxicephalus cf. adspersus BMNH 2018.5789 SL2083 MK464307 Sioma Ngwezi NP -16.89873 23.59847 1009

Pyxicephalus cf. adspersus BMNH 2018.5790 SL2084 MK464306 Sioma Ngwezi NP -16.89873 23.59847 1009

Pyxicephalus cf. adspersus BMNH 2018.5791 SL2097 MK464305 Sioma Ngwezi NP -16.89873 23.59847 1009

Tomopterna marmorata BMNH 2018.5792 SL2070 MK464287 Maramba Lodge, Livingstone -17.89120 25.85821 900

Tomopterna sp. BMNH 2018.5793 SL2078 MK464286 Sioma Ngwezi NP -16.89873 23.59847 1009

Tomopterna sp. BMNH 2018.5794 SL2085 MK464285 Sioma Ngwezi NP -16.89873 23.59847 1009

Tomopterna sp. BMNH 2018.5795 SL2086 MK464284 Sioma Ngwezi NP -16.89873 23.59847 1009

Tomopterna sp. BMNH 2018.5796 SL2092 MK464283 Sioma Ngwezi NP -16.89873 23.59847 1009

Tomopterna sp. BMNH 2018.5797 SL2093 MK464282 Sioma Ngwezi NP -16.89873 23.59847 1009

RHACOPHORIDAE

Chiromantis xerampelina BMNH 2018.5798 SL 2036 MK464456 Mayukuyuku, Kafue NP -14.91533 26.06311 1012

REPTILIA: SQUAMATA

AGAMIDAE

Agama armata BMNH 2018.2751 SL2253 MK464483 Negonye Falls Camp -16.66139 23.57280 929

Amphib. Reptile Conserv. 26 August 2019 | Volume 13 | Number 2 | e181

Bittencourt-Silva

Appendix 1 (continued). List of amphibians and reptiles found in western Zambia, including species vouchers, GenBank accession

numbers, and locality information. Museum acronym: BMNH — Natural History Museum, London, United Kingdom. GPS datum WGS-84.

Species Voucher ID Field ID GenBank Locality Latitude Longitude Altitude

CHAMAELEONIDAE

Chamaeleo dilepis BMNH 2018.2755 SL2099 MK464458 Chavuma FR -13.07006 22.92880 1073

Chamaeleo dilepis BMNH 2018.2756 SL2100 MK464457. Chavuma FR -13.07006 22.92880 1073

COLUBRIDAE

Crotaphopeltis hotamboeia BMNH 2018.2773 SL2247 MK464455__ Hillwood Farm -11.26690 24.31666 1308

Crotaphopeltis hotamboeia BMNH 2018.2776 SL2272 Itezhi-Tezhi, Kafue NP -15.77340 26.01151 1036

Crotaphopeltis hotamboeia BMNH 2018.2777. SL2258 MK464454_ — Sioma Ngwezi NP -16.89873 23.59847 1009

i a Se BMNH 2018.2772 SL2249 = MK464304 Hillwood Farm -11.26690 _24,31666 1308

Philothamnus hoplogaster | BMNH 2018.2775 SL2277 + MK464398 _ Nanzila Plains, Kafue NP -16.28138 25.91676 1032

Thelotornis kirtlandii BMNH 2018.2760 SL 2182 Hillwood Farm -11.26690 24.31666 1308

GEKKONIDAE

Hemidactylus mabouia BMNH 2018.2740 SL 2264 Itezhi-Tezhi, Kafue NP -15.77340 26.01151 1036

Hemidactylus mabouia BMNH 2018.2742 $SL2263 MK464452 __Itezhi-Tezhi, Kafue NP -15.77340 2601151 1036

Hemidactylus mabouia BMNH 2018.2741 SL2276 MK464453 —Nanzila Plains, Kafue NP -16.28138 25.91676 1032

Lygodactylus angolensis BMNH 2018.2766 SL2213 MK464405— Nkwaji -11.52743 _ 24,53532 1343

Lygodactylus angolensis BMNH 2018.2767 SL2187 MK464404— Nkwaji -11,60592 _24.55448 1244

Lygodactylus chobiensis BMNH 2018.2743. SL2267 MK464403 __Itezhi-Tezhi, Kafue NP -15.77340 2601151 1036

Lygodactylus chobiensis BMNH 2018.2744 SL 2268 Itezhi-Tezhi, Kafue NP -15.77340 26.01151 1036

Pachydactylus punctatus | BMNH 2018.2757 SL2265 = MK464400 __Itezhi-Tezhi, Kafue NP -15.77340 2601151 1036

Pachydactylus punctatus BMNH 2018.2758 SL 2266 Itezhi-Tezhi, Kafue NP -15.77340 26.01151 1036

Pachydactylus punctatus BMNH 2018.2759 SL 2269 Itezhi-Tezhi, Kafue NP -15.77340 26.01151 1036

GERRHOSAURIDAE

Gerrhosaurus bulsi BMNH 2018.2754. SL2181 Hillwood Farm -11.26690 24.31666 1308

LACERTIDAE

Ichnotropis capensis BMNH 2018.2746 SL2103 MK464417. Chavuma FR -13.07006 22.92880 1073

Ichnotropis capensis BMNH 2018.2747. SL2260 MK464416 ~~ Lukwakwa -12.66084 24.43697 1063

Ichnotropis capensis BMNH 2018.2749 = SL 2259 Lukwakwa -12.66084 24.43697 1063

Ichnotropis capensis BMNH 2018.2748 SL 2274 Nanzila Plains, Kafue NP -16.28138 25.91676 1032

Ichnotropis capensis BMNH 2018.2750 SL2273. MK464415_—_Nanzila Plains, Kafue NP -16.28138 25.91676 1032

Ichnotropis capensis BMNH 2018.2745 SL2256 MK464418 — Sioma Ngwezi NP -16.89873 23.59847 1009

Meroles squamulosus BMNH 2018.2753. SL2275 MK464401 _—Nanzila Plains, Kafue NP -16.28138 25.91676 1032

Meroles squamulosus BMNH 2018.2752 SL2257 MK464402 —‘ Sioma Ngwezi NP -16.89873 23.59847 1009

LAMPROPHIIDAE

Atractaspis congica BMNH 2018.2274. SL2160 | MK464459 Hillwood Farm -11.27444 24,32444 1416

SCINCIDAE

Typhlacontias rohani BMNH 2018.2761 SL2254. MK464275 nee ate ee erSioMma 1476966053. 83356743 999

Panaspis cf. wahlbergi BMNH 2018.2738 SL2101 MK464399 Chavuma FR -13.07006 — 22.92880 1073

Panaspis cf. wahlbergi BMNH 2018.2739 SL 2104 Chavuma FR -13.07006 22.92880 1073

ae oe BMNH 2018.2762 SL2270 MK464281 _ Itezhi-Tezhi, Kafue NP -15.77340 26.0115] 1036

ph eS BMNH 2018.2763 SL 2032 Mayukuyuku, Kafue NP -14.91533 26.0631 1012

Amphib. Reptile Conserv. 27 August 2019 | Volume 13 | Number 2 | e181

Herpetological survey of western Zambia

Appendix 1 (continued). List of amphibians and reptiles found in western Zambia, including species vouchers, GenBank accession

numbers, and locality information. Museum acronym: BMNH — Natural History Museum, London, United Kingdom. GPS datum WGS-84.

Species Voucher ID Field ID GenBank Locality Latitude Longitude Altitude

Praclylepicch BMNH 2018.2765 SL2013 MK464280 Mayukuyuku, Kafue NP -14.91533 2606311 1012

albopunctata

Trachylepis damarana BMNH 2018.2764 SL2255 | MK464279 Rae Cs OFSHOMAS 9 1666953 2956743 999

Trachylepis wahlbergii BMNH 2018.2769 SL2271 MK464278 __Itezhi-Tezhi, Kafue NP -15.77340 26.0115] 1036

Trachylepis wahlbergii BMNH 2018.2768 SL 2262 Lukwakwa -12.66084 24.43697 1063

Trachylepis wahlbergii BMNH 2018.2770 SL2261 MK464277.— Lukwakwa -12.66084 24.43697 1063

Trachylepis wahlbergii BMNH 2018.2771 SL2188 | MK464276 = Nkwaji -11.60592 24,.55448 1244

REPTILIA: Testudines

TESTUDINIDAE

Kinixys spekii No voucher Ne Nkwaji -11.53906 24.55262 1336

voucher

Amphib. Reptile Conserv. 28 August 2019 | Volume 13 | Number 2 | e181

Amphibian & Reptile Conservation

13(2) [Special Section]: 29-41 (e182).

Official journal website:

amphibian-reptile-conservation.org

Hiding in the bushes for 110 years: rediscovery of an

iconic Angolan gecko (Afrogecko ansorgii Boulenger,

1907, Sauria: Gekkonidae)

12.5.*xPedro Vaz Pinto, ‘Luis Verissimo, and **William R. Branch

'Fundagao Kissama, Rua 60 Casa 560, Lar do Patriota, Luanda, ANGOLA *CIBIO/InBio — Centro de Investigacao em Biodiversidade e Recursos

Genéticos, Universidade do Porto, Campus Agrario de Vairdo, 4485-661, Universidade do Porto, PORTUGAL °Port Elizabeth Museum (Bayworlad),

P.O. Box 13147, Humewood 6013, SOUTH AFRICA *Research Associate, Department of Zoology, P.O. Box 77000, Nelson Mandela University, Port

Elizabeth 6031, SOUTH AFRICA (deceased 14 October 2018) °>TwinLab CIBIO-ISCED, Instituto Superior de Ciéncias da Educagdo da Huila, Rua

Sarmento Rodrigues s/n, Lubango, ANGOLA

Abstract.—Boulenger (1907) described a new gecko ‘Phyllodactylus’ ansorgii based on two adult females

from ‘Maconjo, Benguella, Angola,’ but subsequent taxonomic reviews of leaf-toed geckos ascribed southern

African lineages to new genera and this species has since been tentatively placed under ‘Afrogecko.’ For over

110 years the type locality remained a mystery, and the gecko became a lost icon of Angolan herpetology.

Early searches for the gecko were confounded by misinterpretation of the type locality ‘Maconjo,’ which it is

now evident was confused with a toponym that is a well-known historical locality. Following the discovery of

new material in coastal Benguela, an examination of historical documents and cartographic material allowed

the original collecting area for the type material to be identified. Specific surveys in this area resulted in the

collection of topotypic material and the recording of behavioral observations and notes on the ecology of the

species. Afrogecko ansorgiiis a slender, gracile, and arboreal gecko that inhabits small bushy trees, particularly

blackthorn (Senegalia mellifera subsp. detinens), in the arid coastal scrubland of the Benguela coastal region.

All A. ansorgii have been found in or nearby blackthorn in which the activity of termites (Kalotermitidae) has

created hollow stems in which the geckos shelter. The type locality for Phyllodactylus (= Afrogecko) ansorgii

Boulenger, 1907 and Mabuia (= Trachylepis) laevis Boulenger, 1907, both described at the same time from

‘Maconjo, Benguella’ is accordingly restricted, and topotypic material for the latter species was also obtained.

Key Words. Ansorge, blackthorn, Kaokoveld, Maconjo, Reptilia, type locality, Trachylepis laevis

Resumo.—Boulenger (1907) descreveu a nova osga ‘Phyllodactylus’ ansorgii a partir de duas fémeas

oriundas de ‘Maconjo, Benguella, Angola,’ mas subsequentes revisOes taxonomicas das osgas-de-dedos-de-

folha fez corrresponder a novos géneros as linhagens da Africa austral e desde entao esta espécie tem sido

tentativamente incluida em ‘Afrogecko.’ Durante mais de 110 anos a localidade tipica permaneceu um misteério,

e a osga tornou-se um icone perdido da herpetologia Angolana. Buscas iniciais pela osga ficaram baralhadas

devido a uma ma interpretacao da localidade tipica ‘Maconjo,’ que é hoje evidente ter sido confundida com

um toponimo que é uma localidade historica bem conhecida. Apos a descoberta de novo material na regiao

costeira de Benguela, a despistagem de documentos historicos e material cartografico permitiu identificar

a area de colheita original do material tipico. Levantamentos especificos nesta area resultaram na colheita

de material topotipico e no registo de observagoes comportamentais e notas sobre a ecologia da espécie. O

Afrogecko ansorgii € uma osga delgada, delicada e arboricola que habita pequenas arvores de porte arbustivo,

particularmente espinheiras (Senegalia mellifera subsp. detinens), nas estepes aridas arbustivas da regiao

costeira de Benguela. Todos os A. ansorgii foram encontrados nestas espinheiras ou na sua vizinhanga, e onde

a actividade de termitas (Kalotermitidae) gerou talos ocos onde as osgas se abrigam. A localidade tipica para o

Phyllodactylus (= Afrogecko) ansorgii Boulenger 1907 e para o Mabuia (= Trachylepis) laevis Boulenger 1907,

ambos descritos na mesma altura para ‘Maconjo, Benguella’, é desta forma restringida, e material topotipico

da ultima espécie foi tambem obtido.

Palavras-chave: Reptilia, localidade tipica, Maconjo, Ansorge, espinheira, Kaokoveld, Trachylepis laevis

Citation: Vaz Pinto P, Luis Verissimo L, Branch WR. 2019. Hiding in the bushes for 110 years: rediscovery of an iconic Angolan gecko (Afrogecko

ansorgii Boulenger, 1907, Sauria: Gekkonidae). Amphibian & Reptile Conservation 13(2) [Special Section]: 29-41 (e182).

tion 4.0 International (CC BY 4.0): https://creativecommons.org/licenses/by/4.0/], which permits unrestricted use, distribution, and reproduction in any

medium, provided the original author and source are credited. The official and authorized publication credit sources, which will be duly enforced, are

as follows: official journal title Amphibian & Reptile Conservation; official journal website: amphibian-reptile-conservation.org.

Submitted: 12 March 2019; Accepted: 14 March 2019; Published: 11 August 2019

Correspondence. * pedrovazpinto@gmail.com

Amphib. Reptile Conserv. 29 August 2019 | Volume 13 | Number 2 | e182

Rediscovery of Afrogecko ansorgii in Angola

Introduction

The biodiversity of Angola, and particularly that of the

herpetofauna, is acknowledged to be inadequately known

(Branch 2016; Huntley and Ferrand 2019). This was

not always the case, and like many countries in Africa

the early phase of colonial exploration and settlement

resulted in the discovery of numerous novel animals.

For almost 50 years (1866-1915) these collections were

sent to European museums for study and description, and

they stimulated much scientific interest. Early studies on

the Angolan herpetofauna were numerous, particularly

those of José Vicente Barbosa du Bocage, the father of

Angolan herpetology. His monographic Herpétologie

d’Angola et du Congo (Bocage 1895) remained for

over a century the definitive synthesis of the country’s

herpetofauna, until the recent publication of a historic

herpetological atlas (Marques et al. 2018). Currently,

studies on the herpetofauna of Angola are entering a

new phase following the closure of protracted hostilities

at the end of the colonial era. Recent field surveys (e.g.,

Huntley 2009; Ernst et al. 2014; Huntley and Francisco

2015; Ceriaco et al. 2016, 2018; Conradie et al. 2016)

have uncovered new species of amphibians (Conradie

et al. 2012a, 2013; Ceriaco et al. 2018) and reptiles

(Conradie et al. 2012b; Stanley et al. 2016). However,

a number of important historical species described

during the colonial period still remain known from only

their original description, e.g., Sepsina copei Bocage,

1866 and Cordylus angolensis (Bocage 1895), or only

a few subsequent specimens, e.g., Monopeltis luandae

Gans 1976 (Branch et al. 2018), Psammophis ansorgii

Boulenger 1905, and Psammophylax ocellatus Bocage

1873 (Branch et al. 2019a). Tragically, much of the

historical material studied by Bocage, including almost

all of his type material, was lost in the fire that destroyed

the Museu Bocage collections in 1978.

Perhaps the most iconic of the ‘lost’ Angolan species

is Ansorge’s Leaf-toed Gecko, which was described by

Boulenger (1907) as Phyllodactylus ansorgii, over 110

years ago. Leaf-toed geckos have a globally wide-ranging

distribution and all of them used to be included within

Phyllodactylus until a comprehensive review performed

by Bauer (1997) ascribed the southern African species to

three new genera. The genus Afrogecko initially included

two species found in South Africa, A. porphyreus, and

A. swartbergensis, and two Angolan leaf-toed geckos, A.

plumicaudus and A. ansorgii (Bauer 1997; Haacke 2008).

However, more recent phylogenetic studies revealed

deeply independent evolutionary histories among these

lineages, leading A. swartgergensis and A. plumicaudus

to be transferred to the new monotypic genera Ramigekko

and Kolekanos, respectively, while due to the lack of

available material ansorgii was provisionally maintained

within Afrogecko (Heinicke et al. 2014).

The species description was based on two female

specimens from Maconjo, Benguella (now Benguela)

collected by Dr. W.J. Ansorge on one of his Angolan

journeys, between 1903 and 1906, and the types, BMNH

1946.8.24.52—53, were deposited in the Natural History

Museum London (United Kingdom). The exact locality

Amphib. Reptile Conserv.

of Ansorge’s Maconjo has caused confusion, and

erroneous interpretation of its whereabouts probably

delayed the species’ rediscovery. Moreover, the species

is now known to live in an unusual habitat, and this may

have also led to it being overlooked. Crawford-Cabral

and Mesquitela (1989) summarized all known localities

for Angolan terrestrial vertebrates and considered

Maconjo a variant spelling of Maconge, an old farm

situated in Namibe Province, formerly Mossamedes (=

Mocamedes) District (15°01’S, 13°12’E, 700 m asl).

Maconjo, Mossamedes features as a collecting locality

for reptiles in the 19" century for the famous Portuguese

collector José de Anchieta (de Andrade 1985), and in his

monograph Bocage (1895) recorded numerous reptiles

from Maconjo, Mossamedes, including: the terrapin

Pelomedusa galeata (= P. subrufa); the lacertids Nucras

tessellata and Eremias lugubris (= Heliobolus lugubris),

the rupicolous skink Mabuia chimbana (= Trachylepis

chimbana);, the python Python natalensis, the snakes

Prosymna_ frontalis (= P. angolensis, fide Broadley

1980), Psammophylax nototaenia (= Hemirhaggheris

viperina, fide Broadley 2000), Psammophis sibilans (=

P. subtaeniatus), Elapsoidea guentheri var. semiannulata

(= E. s. semiannulata), and Causus resimus. Many of

these have been subsequently recorded in the region

(data not shown).

A farm and stream at the base of the Humpata plateau

near Leba Pass are called Maconjo on old maps and lie

in the vicinity of the old Portuguese fort of Capangombe,

which dates from the mid-nineteenth century and marks

the route for some of the earliest inland incursions of

colonists in southern Angola. This locality was then

included in the District of Mossamedes. However,

Boulenger referred the gecko’s type locality to “Maconjo,

Benguella,’ placing it in Benguela instead of Mossamedes

District. This suggests either that it was ascribed by

Ansorge to the wrong district, or that there could exist

two different sites bearing the name Maconjo. It should

be noted that mistaking Mossamedes for Benguela would

have been unlikely in the early 20" century, when both

districts were well established and had been for some

time. They have also remained as separate administrative

entities ever since. Yet this incongruence seems to have

gone unnoticed, and it appears that most subsequent

researchers have considered there was only one Maconjo,

and the references to different provinces, Mossamedes (=

Namibe) or Benguela, were either ignored or taken as

synonyms (Bauer etal. 1997; Heinicke et al. 2014; Ceriaco

et al. 2016; Uetz et al. 2018). The accepted wisdom that

there was only one Maconjo, and that it was situated

below the escarpment in the current Namibe Province,

resulted in numerous unsuccessful searches for the

gecko around Capangombe by various researchers from

1974 to 2016. More recently, Maconjo was tentatively

synonymized with another well-known collecting site

in southwestern Angola ‘Fazenda Mucungo’ (Marques

et al. 2018), however, no further details were provided

to justify the new locality. Mucungo also lies on the

coastal plain of Namibe and therefore cannot resolve the

geographical discrepancies.

Another species from ‘Maconjo’ that Boulenger

(1907) described from the same Ansorge collection

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Vaz Pinto et al.

was the small rupicolous skink Mabuia (= Trachylepis)

laevis, commonly called the Angolan Blue-tailed Skink

(Tf. laevis). It is rupicolous, sheltering in thin cracks in

hard, fractured rocks and has numerous morphological

adaptations to this habitat (Paluh and Bauer 2017).

Despite its conspicuous coloration, the species has

only rarely been collected in Angola after the original

description (Boulenger 1907). Laurent (1964) recorded

a male from Munhino, 50 km W of Lubango, and

Hellmich (1957) reported a problematic specimen from

Piri-Dembos (= Piri) that should be treated with caution

(Laurent 1964; Ceriaco et al. 2016). The Angolan Blue-

tailed Skink was recently recorded from granite outcrops

50 km east of Namibe (W.R. Branch, pers. comm.), and

just north of Tambor near Iona National Park (Ceriaco et

al. 2016), and additional material has been collected in

northern Namibe Province, at Chapéu Armado, Serra da

Neve, and Lola by the authors of the current study. Still,

the most significant collections of 7’ /aevis were made

by Wulf Haacke during an expedition to southwestern

Angola in 1971, when he collected 18 specimens from

Tambor and other localities in Namibe Province (all

deposited in the Ditsong Natural History Museum,

Pretoria, South Africa). In addition, and importantly, he

collected the first records since Ansorge from Benguela

Province, with material from 35 km south of Dombe

Grande and 53 km south of Benguela.

Until now, no additional material for A. ansorgii had

ever been collected, despite searches by experienced

herpetologists in the 1970s and after 2009. In this paper

we report on finding the species and obtaining new

material that includes the first male specimens. The

type locality is here confidently restricted, and topotypic

material was collected for both A. ansorgii and T. laevis.

In addition, by collecting the gecko on a second site, we

extend its known range by about 100 km. Behavioral

and habitat observations were also recorded, and provide

useful insights into their unique ecological requirements.

Materials and Methods

To investigate local toponyms and locate the type

locality, historical cartographic material was thoroughly

examined and, in the absence of available diaries for Dr.

Ansorge’s Angolan journeys, we consulted a remarkable

manuscript by the American ornithologist James Chapin

which provides a detailed summary of Ansorge’s

itineraries in Angola (Chapin, undated).

After first encountering A. ansorgii, we conducted

a series of additional field trips in search of the gecko

between November 2016 and June 2018, to five different

sites, including the restricted type locality and additional

locations where we looked for their presence. The surveys

included mostly nocturnal searches, and daytime habitat

observations. A total of 23 specimens of A. ansorgii

were collected in two sites, including the restricted type

locality. All specimens were photographed in life, and

subsequently preserved in formalin. Collected specimens

were deposited at Port Elizabeth Museum (PEM R23907—

23916), at ISCED — Instituto Superior de Ciéncias da

Educacdo da Huila (NB603-605; NB822-—826) and at

Kissama Foundation (KFH0007—0010). Additionally,

Amphib. Reptile Conserv.

behavioral observations were also recorded by locating

the geckos at night with flashlights and observing their

foraging habits, and by confirming their daytime shelters.

Reptile species lists were compiled for each site visited,

and topotypic material was also secured for 7: /aevis,

with one specimen collected and deposited at Kissama

Foundation collection (FKH0018).

Results

Type locality: From Chapin’s summary it is apparent

that Ansorge visited ‘Makonjo, Benguella’ on 7 July

1904. Chapin used the spelling ‘Makonjo’ instead of

following Boulenger’s ‘Maconjo,’ but the use of a ‘k’

or ‘c’ 1s often interchangeable when applied to Angolan

toponymy. To avoid continued confusion with toponyms

we hereafter keep the spelling as Maconjo and add the

district/province name when relevant. Ansorge left the

locality of “‘Huxe, Benguella’ (= Uche, 12°44’S, 13°21’E,

250 m asl) on the previous day on his way to Benguela’s

sandpits and the town of Catengue (13°02’S, 13°44’E,

550 m asl), which he reached on 11 July 1904. This route,

depicted on a map included in the manuscript, places the

locality of Maconjo clearly in Benguela as opposed to

Namibe’s coastal plain, and within a few days travel on

foot from Benguela town. In addition, there are no other

records of localities of similar spelling in the manuscript,

and no suggestion that Ansorge ever visited Capangombe

region. Moreover, based on Chapin’s document it appears

that Ansorge never descended the escarpment from the

Humpata plateau, and although he referred to his 1906

route south of Caconda as being in Mossamedes District,

Chapin failed to recognize the then newly created Huila

District. As a result, there is no evidence to suggest

that Ansorge even collected in the former Mossamedes

District as it was defined in his time, and which broadly

corresponds to the current Namibe Province.

Comparing Chapin’s notes with cartographic material

allowed the reconstruction of Ansorge’s probable route

(Fig. 1) and the identification along it of a region by

the name of ‘Conjo’ (12°53’S, 13°24’E) in Benguela

Province. This area, approximately 20 km south of Uche

and on the route towards Catengue, lies between 300

and 450 m asl, and includes two small north-flowing dry

sandy streams named respectively as “Conjo’ and ‘Conjo

Pequeno,’ both tributaries to another ephemeral stream

named ‘Cocumba.’ The region is in Benguela’s coastal

plain and would be realistically within one-day’s reach

from Uche on foot, and enroute to Catengue, making

it extremely likely that it corresponds to the true type

locality of “Maconjo, Benguela’ as recorded by Ansorge

(in Boulenger 1907). Interestingly, in the local language

Conjo means spoor and the prefix ‘Ma-’ is commonly

used to signify plurality, so the toponymy Maconjo

can be appropriately interpreted as ‘the site of various

Conjos’ or ‘site of many spoor.’

On the basis of Chapin’s notes and cartographic

material we here restrict the type locality of Phyllodactylus

(= Afrogecko) ansorgii and Mabuia (= Trachylepis) laevis,

both described by Boulenger (1907) from ‘Maconjo,

Benguella,’ to the vicinity of the streams Conjo, Conjo

Pequeno, and Cocumba (12°52’S, 13°21’E, 355 m asl),

August 2019 | Volume 13 | Number 2 | e182

Rediscovery of Afrogecko ansorgii in Angola

* Type locality - Maconjo, Benguela

—-=—- Ansorge Routes, 1904 to1906, in SW Angola

@ Newsite - sta. Maria

@ = Additional surveyed sites

—--- Angola Administratrive Districts in 1905

Fig. 1. Travel routes in Angola by Dr. Ansorge between 1904

and 1906, as per Chapin’s manuscript. Former district (broadly

corresponding to current provinces) borders are shown, as well

as collecting sites and other relevant localities.

20 km south of Uche, Benguela Province, Angola (Fig.

1). Both species, Afrogecko ansorgii and Trachylepis

laevis, were collected at this site (see below).

Rediscovery: The gecko was first rediscovered 135 km

southwest of Benguela town on 12 November 2016. The

site is located about three km from the Lucira-Benguela

road, at the head of a valley leading down to Cape Sta

Maria, Benguela Province (13°31’S, 12°38’E, 288 m

asl). The locality is ca. 185 km northwest of Maconjo,

Namibe, and ca. 100 km southwest of the type locality

in Benguela Province (Fig. 1). A total of 10 specimens

were collected at night whilst foraging in the branches of

thorny bushes (Fig. 2), mainly of blackthorn Senegalia

mellifera detinens. Subsequently, three additional

specimens were collected, and nine others observed in

the same habitat and under similar circumstances, at two

other nearby sites surveyed five km and three km to the

northwest, on 12—13 July 2017 and 11-12 August 2017,

respectively. Most specimens were located at night on

bushes situated at the base or mid-slopes of granitic hills,

and although a few were observed in various species of

small trees, the majority were found among the spiny

branches of blackthorn bushes. One specimen was found

during the day sheltering inside a hollow blackthorn

branch (Fig. 3A).

After the cartographic identification of the potentially

true type locality, specific visits to the Conjo area to

search for geckos were made on 25—26 November 2017

Amphib. Reptile Conserv.

and 5 December 2017. The region surveyed was located

approximately half-way between the towns of Benguela

(35 km) and Catengue (47 km) and five km south of the

small village of Talamajamba. As it shared some features

with sites at Sta Maria, although lacking in rocky habitat

suitable for Trachylepis laevis, we first focused on a site

located on a sand road that crosses the Cocumba dry

stream and runs a few hundred meters parallel to the east

of Conjo Pequeno (12°54’3S, 13°24’E, 395 m asl). The

searches resulted in our obtaining topotypic material and

further observations. Broadening the search, additional

visits on 12-13 March 2018 and 5 June 2018, focused

on an area around three isolated granite outcrops present

along the stream Cocumba and a mere 500 m west of the

confluence with stream Conjo (12°52’S, 13°21’E, 355 m

asl). Although the Cocumba is a dry stream on a semi-

desert environment, local pastoralists have long dug

artisanal wells to gain access to water on the river bed

at the base of the granite outcrops. The presence of these

wells on the confluence of Conjo and Cocumba streams

offers a realistic scenario for the site ‘Maconjo’ to have

been used as a stopover locality between Benguela

and Catengue by Ansorge in 1904. Here, additional

specimens of A. ansorgii were found in nearby bushes,

and a specimen of 7: /aevis was collected in the granite

outcrops, constituting the first topotypic material for this

species.

At Maconjo, a total of 10 specimens of A. ansorgii

were collected and 14 additional individuals were

observed foraging at night in bushes over the four visits,

mostly found in Senegalia mellifera detinens, but also

in Terminalia prunioides, Commiphora cf. africana, and

Salvadora persica, and once on a dry grass stem. Most

geckos were spotted in the early evening (approximately

between 19h00 and 21h00) in small branches 1-2 m

above ground. One specimen was observed partially

exiting a hollow blackthorn branch shortly after sunset,

as exemplified in a photograph (Fig. 3B).

Three additional coastal sites (Fig. 1) were also

surveyed in search of the geckos, one was an intermediate

location between Maconjo and Sta Maria, at Chimalavera

Regional Park (12°50’S, 13°10’E, 255 m asl), on a sandy

plateau over an east-facing limestone ridge, Benguela

Province; another ca. 35 km south of Maconjo and near

the Coporolo River (13°12’S, 13°25’E, 380 m asl); and

one further south at 17 km W of Chicambi Village, on

sandy flats with a granite ridge, in Namibe Province

(13°55’S, 12°41’E, 531 m asl). In these sites the habitat

was similar to that at Cape Sta Maria, with numerous

Senegalia mellifera detinens bushes on sandy soil and

with adjacent rock outcrops. However, in all these three

cases the blackthorn bushes lacked hollow branches and

no geckos were observed during nocturnal searches.

Habitat observations: On the vegetation map of

Angola, Barbosa (1970) defines a very extensive region

as semi-desert coastal steppe, starting as a narrow coastal

strip around 11° south, then becoming broader inland

towards the southern border with Namibia. However,

this is a very wide and rough classification and includes a

transition from dry spiny savannahs dominated by acacias

(Senegalia spp.) to mopane woodlands characterized

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Vaz Pinto et al.

Fig. 2. Afrogecko ansorgii: adult male, PEM R23907 (in life). (A) Whole body on small branch of Senegalia mellifera detinens with

epiphytic lichen. (B) Close up of head of same adult male. (C) Afrogecko ansorgii: adult female, PEM R23912 (in life) on small

branch of Senegalia mellifera detinens. (D) Close up of head of same adult female. Note elongate, subcylindrical body, relatively

short cylindrical tail that is shorter than SVL and partially prehensile, long toes with expanded terminal scansors, predominantly

dark brown dorsal coloration, and dorsolateral series of irregular pale blotches that extend on to the tail.

by the dominance of Colophospermum mopane. Still,

he refers to the arid regions near Benguela as being

characterized by an abundance of Senegalia mellifera

detinens, Senegalia spp., Salvadora persica, Euphorbia

spp., and Boscia spp. (Barbosa 1970) which likely

correspond to the more relevant areas for Ansorge’s

Gecko.

At Maconjo, Benguela, the restricted type locality,

the terrain is relatively flat, with sandy soils, irregularly

interspersed with small granitic rocks, and with a few

isolated granite outcrops at the Cocumba stream (Fig.

4A). The vegetation was relatively dense and diverse, the

trees often in close proximity or touching, but with low

canopies and not growing larger than the size of a bush

(Fig. 4B). The dominant tree/bush species were Senegalia

mellifera detinens and Terminalia prunioides, but other

species also found to be common were Commiphora cf.

africana, Boscia microphylla, Vachellia reficiens, and

Salvadora persica. Succulents were also recorded, notably

the conspicuous Aloe littoralis, plus a few Hoodia cf.

parviflora and the invasive Opuntia ficus-indica.

The Sta Maria site is situated at a slightly lower

elevation, being closer to the coast and further south than

the type locality, and in a more arid coastal environment.

It possibly receives lower rainfall, but this may be

Amphib. Reptile Conserv.

compensated for by the higher incidence of coastal fog

which is a feature of the whole Namib Desert coastline

(Cermak 2012). The effect of regular fogs, which may

extend well inland, is evident in the local abundance of

lichens that often cover the branches of bushes (Figs. 2A,

4C). At this locality the vegetation was much sparser than

at Maconjo, Benguela, particularly in the sandy valleys,

but the topography was often much more varied with

large granite boulders and steep granitic hills (Fig. 4D).

Senegalia mellifera detinens was the most abundant tree,

while other common species recorded were Salvadora

persica, Commiphora multijuga, Phaeoptilum spinosum,

and Jerminalia prunioides.

A striking find that appears positively linked with the

occurrence of the gecko is the existence in both sites of

abundant hollow branches in Senegalia mellifera detinens

caused by the activity of termites (Kalotermitidae) [Fig.

5]. By eating the heartwood of dead branches and leaving

the outer wood and bark intact, the termites hollow out

branches, making small exit holes at the branching nodes.

This activity creates ideal daytime shelters for Afrogecko

ansorgii, with multiple entry and exit points. These

hollow branches subsequently break off and accumulate

below the tree, where they may still be used as refugia

by the geckos.

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Rediscovery of Afrogecko ansorgii in Angola

Table 1. Variation observed for Afrogecko ansorgii, comparing the type series from Maconjo, with new series obtained in Maconjo

and Sta Maria. Continuous measurements in mm (mean and range) for TL= total length, SVL= snout-vent length and Tail. Meristic

and qualitative variation (median and range) is given for number of precloacal pores and cloacal spurs.

TL SVL Tail Pores Spurs

Type Series — Maconjo

Females (n=2) 7) 45 30 0 na

New Series — Maconjo

Females (n=5)* Ts) 43.9 S03 0 ]

(71.4-79.0) (41.3-45.8) (30.1-33.2) 0 l

Males (n=4) 65 38.2 26.8 8 2

(57.8-68.8) (34.4—40.6) 23.4—29.0) 8-9 2

Juvenile Female (n=1) 68 38.6 29.4 0 1

New series — Sta Maria

Females (n=5)** 63.3 39.4 23.8 0 0

(56.2—69.3) (34.7—42.9) (22.0—26.4) 0) 0

Males (n=6)** 60 35.7 24.2

(53.2—63.5) (34.4-38.0) (18.8—27.5) fis 152

Juvenile Female (n=1) 54.4 B23 piers 0 0

* For Tail and TL measurements n=3 due to truncated tails; ** For Tail and TL measurements n=4 due to truncated tails.

Natural history: The geckos were mostly first seen lying

flat or moving slowly on thin branches in the early hours

of the evening. Nevertheless, they proved to be agile

when disturbed, often running along the branches or even

jumping between twigs. The tail was kept low most of

the time and appeared to be semi-prehensile. Based on

our observations they retreat to safety inside the hollow

branches of Senegalia mellifera detinens during the day,

and emerge soon after dark to forage, mostly on thin

branches but not restricted to the acacias. They likely

avoid the ground and no specimen was found walking

on soil or rock, but their agility allows them to move

across neighboring trees, particularly where there is a

high density of bushes, as at Maconjo, or where canopy

contact is facilitated by uneven and steep slopes as in Sta

Maria. A nocturnal arboreal foraging behavior has also

Fig. 3. Afrogecko ansorgii in its biotope, the termite-hollowed, thin branches of Senegalia mellifera detinens. ( )

hollow blackthorn twig. (B) Specimen caught trapped inside a blackthorn branch.

Amphib. Reptile Conserv.

been recorded for another Angolan endemic gecko, the

possibly closely related species Kolekanos plumicaudus,

although the latter retreats into rock crevices for shelter

(Agarwal et al. 2017).

The geckos probably prey on small insects found on

the outer branches of bushes, and it is possible that they

also feed on alates of the same termites that provide them

with the sheltering habitat, although this could not be

confirmed as gut content analyses were not performed.

The vast hollow branches with narrow entrances likely

preclude many snakes from preying on adults and eggs.

In any case, one specimen of Psammophis leopardinus

was collected, while actively moving at night among the

outer branches of a blackthorn bush at Maconjo (Fig.

6A), strongly suggesting that it was hunting A. ansorgii.

At both localities the gecko MHemidactylus cf.

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par)

August 2019 | Volume 13 | Number 2 | e182

Vaz Pinto et al.

Fig. 4. Habitat of Afrogecko ansorgii. (A) Isolated granite outcrop beside the dry Cocumba stream near its confluence with Conjo

Stream (12°52’S, 13°21’E, 355 m asl), the correct site of Boulenger’s type locality “Maconjo’ for Phyllodactylus ansorgii and

Mabouia laevis. (B) Spiny savanna of Senegalia mellifera detinens with Terminalia prunioides and Aloe littoralis at the type

locality. (C) Scattered blackthorn bushes subject to thick morning fog and covered with lichen within which Afrogecko ansorgii was

rediscovered. (D) Aerial view of dry spiny savannah at the head of a valley leading down to Cape Sta Maria, Benguela Province

(13°31’S, 12°38’E, 288 m asl).

longicephalus was collected in syntopy in_ the

same bushes, although usually foraging on thicker

branches and closer to the ground. Other rupicolous

reptile species on the surrounding rocks on both sites

included Pachydactylus caraculicus, Chondrodactylus

fitzsimonsi, C. pullitzerae, Trachylepis laevis (Fig.

6B), 7) sulcata ansorgii, Agama planiceps, the diurnal

Rhoptropus cf. benguellensis and R. cf. barnardi; and the

terrestrial species Pachydactylus punctatus, Trachylepis

acutilabris, and Pedioplanis benguellensis. Species

found in Sta Maria but not recorded at Maconjo were

Afroedura cf. bogerti, Pachydactylus cf. oreophilus,

Cordylus namakuiyus, and Hemirhagerrhis viperina,

while Matobosaurus maltzahni, Agama anchietae,

Trachylepis binotata, and Psammophis leopardinus were

found at Maconjo but not at Sta Maria. These records

include northern extension ranges of roughly 200 km or

more for three gekkonids, Chondrodactylus fitzsimonsi,

Pachydactylus caraculicus, and Rhoptropus cf. barnardi,

and for the endemic and recently described Cordylus

namakuiyus (Stanley et al. 2016).

Distribution range: So far, the gecko has only been

found in the two referred localities of Maconjo and Cape

Sta Maria, spanning across over 100 km on Benguela’s

coastal plain. It is likely that the species has a wider

distribution range, where it may be locally common but

Amphib. Reptile Conserv.

irregularly distributed, and dependent on very particular

habitat requirements in association with Senegalia

mellifera detinens and the kalotermitid termites.

New series and morphological variation: The new

material obtained in both localities is consistent with

the original description of Afrogecko ansorgii. The more

slender form and the enlarged precloacal scales well

distinguish this species from A. porphyreus (Daudin

1802), while the larger size, presence of femoral spurs

and a slender semi-prehensile versus a broad flattened

tail clearly separates it from the other Angolan endemic

Leaf-toed Gecko Kolekanos plumicaudus (Haacke 2008).

Boulenger’s description, based on one of two available

adult females, is terse but conforms to that of the era and

is given below, while some measurements, including the

new series, are presented in Table 1. Male specimens

were obtained for the first time, allowing comparisons

between sexes and localities. A detailed re-description

of A. ansorgii is being prepared, which will also explore

phylogenetic relationships and include a taxonomic

review (data not shown). Original description of

Afrogecko ansorgii (Boulenger 1907):

Head rather small, oviform, much longer than broad;

snout not longer than the distance between the eye

and the ear opening, which is small and oval. Body

August 2019 | Volume 13 | Number 2 | e182

Rediscovery of Afrogecko ansorgii in Angola

ti vie

K..

poe

Fig. 5. Kalotermitid termites active on blackthorn bushes at type locality. (A) Hollow branch in tree. (B) Termite soldier and

evidence of wood excavation. (C) Termite and eggs inside branch. (D) Workers active inside hollow branch.

very elongate; limbs moderate. Digits moderately

depressed, with large, subtrapezoid terminal

expansions; eight lamellae under the fourth toe. Head

and body covered with uniform, smooth, flattened

granules, which are larger on the snout and on the

belly. Rostral twice as broad as deep, without cleft

above; symphysial small, a little longer than broad;

ten upper and as many lower labials,; rostral and first

upper labial entering the nostril; no chin-shields. Tail

cylindrical, tapering, covered with uniform, small,

quadrangular, smooth scales. A curved transverse

series of eight or nine enlarged praeanal scales

(indicating praeanal pores in the male?). Pale greyish

brown above, with a series of large whitish spots along

each side of the back; a dark streak on each side of the

head and neck, passing through the eye; upper lip and

lower parts white, with small brown spots.

Two females collected in the type locality were gravid

with two eggs (Fig. 7A). Male specimens are noticeably

smaller than females in snout-vent and total length, with

the difference being more pronounced at the type locality.

As predicted by Boulenger, males have 7—9 precloacal

pores (Fig. 7B), corresponding to the enlarged scales

found on females (Fig. 7C). Of note are the differences

found between the series obtained at Maconjo and

Sta Maria. Specimens from Maconjo proved to be

consistently larger in size and overall measurements,

Amphib. Reptile Conserv.

with females from Sta. Maria of approximately similar

size as males from Maconjo (Table 1). Also striking is the

presence of more cloacal spurs in the series from the type

locality. Although not mentioned in the type description,

we found one cloacal spur in females from Maconjo (Fig.

7A), and two spurs in males from same locality (Fig. 7B).

On the other hand, females from Sta Maria showed no

cloacal spurs, while at the same site all males but one had

one single spur (Table 1).

Discussion

The traditional reliance of gekkonid systematics on

digital morphology is often reflected in the generic

names, with the Leaf-toed Geckos, Phyllodactylus

being one of numerous examples. Until the end of the

20" century, gecko species assigned to Phyllodactylus

had a patchy distribution, spread across five continents.

Increasing awareness of both the antiquity of gekkotan

lineages (Kluge 1987; Conrad and Norell 2007), of

the taxonomic confusion generated by their conflicting

conservative and sometimes convergent morphologies

(Bauer et al. 1997; Gamble et al. 2012), and of the

development and application of molecular phylogenetic

studies (Han et al. 2004; Wiens et al. 2012), have resulted

in a true understanding of gecko antiquity and diversity.

Taxonomic re-arrangements to reflect this evolution and

August 2019 | Volume 13 | Number 2 | e182

Vaz Pinto et al.

i . oo

om 2 we:

a ae a

ve Se et hea,

Fig. 6. (A) Specinieh of Psammophis leopardinus Bimeine at neHe ona blackthorh Bish at Eyal TARE — on A A.

ansorgii. (B) First topotypic material of TZrachylepis laevis collected in over 110 years, FKH0018 in life; note flattened body adapted

to shelter among tight rocky spaces.

cryptic diversity has resulted in the genus Phyllodactylus,

as understood for over 100 years, now being distributed

among the families Diplodactylidae, Gekkonidae, and

Phyllodactylidae and in at least 15 genera! It is thus

not surprising that the generic affinities of Afrogecko

ansorgii remain problematic. When initiating a major

generic revision of phyllodactyline geckos, Bauer et al.

(1997) provisionally assigned ansorgii (then known only

from the types) to the genus Afrogecko, but noted that its

affinities would need re-assessment on discovery of new

material.

Subsequently Haacke (2008) described the new plume-

tailed gecko from the Angolan Namib region that was

again tentatively assigned as Afrogecko plumicaudus in

the absence of genetic studies. The collection of additional

new material by Wulf Haacke allowed its phylogenetic

affinities to be assessed, whereupon it was transferred to

anew gekkonine genus Kolekanos (Heinicke et al. 2014).

Although all the original material was collected under

thin, exfoliating rock slabs, the species was later shown

to forage arboreally (Agarwal et al. 2017). In its (at least)

partial arboreal behavior, K. plumicaudus is similar to

A. ansorgii. Nevertheless, A. ansorgii 1s unique among

southern African leaf-toed geckos, in being exclusively

arboreal and by what appear to be some remarkable

ecological adaptations. The morphological differences

found in specimens obtained at the type locality and Sta

Maria, namely in body size and number of cloacal spurs,

likely reflect local adaptations and may be climatically

driven. Further detailed morphological and genetic

studies are ongoing to confirm the generic placement of

A. ansorgii and explore intraspecific variation (data not

shown).

Southwestern coastal Angola is subject to frequent

advective fogs and relatively cool temperatures, caused

by the cold offshore Benguela current, and experiences

a climatic gradient of increased rainfall from the coast

inland (Huntley 2019). The importance of advective fog

and low clouds in shaping the local arid ecosystems has

been relatively well studied in Namibia, where it has

been estimated to represent up to five times the amount

of precipitation provided by rain (Seeley and Henschel

1998). The frequency of this type of fog is high in coastal

areas, but studies in the central Namib found the amount

of precipitation to be highest around 35—60 km inland

and below 500 m asl, where low strata clouds were

Amphib. Reptile Conserv.

intercepted by local elevation (Lancaster 1984). Satellite

imagery show that advective fog and low clouds penetrate

further inland in the southern and northern sections of

the Namib (Cermak 2012), but relatively little effort has

focused on the role of fog and low clouds in Angola.

Nevertheless, it has been suggested that the incidence of

winter fog and low strata clouds is especially pronounced

between the towns of Namibe and Benguela, leading to

heavy morning dew and allowing for a local abundance

of epiphytic lichens (Huntley 2019). The presence of

these epiphytic lichens was recorded at the site of Sta

Maria, where they often almost covered the branches

of blackthorn bushes. Being close to the sea shore and

situated in an extensive dry valley, the collecting site

at Sta Maria is subject to frequent morning coastal

Fig. 7. (A) Female Afrogecko ansorgii from Maconjo,

FKH0007, gravid with two eggs. Note enlarged precloacal

scales and presence of one cloacal spur. (B) Cloacal region

of male A. ansorgii, FKHO008. Note single chevron row of

nine precloacal pores, hemipenial bulges (as paired light

swellings) and cloacal spurs (small white tubercular scales on

lateral surfaces of tail base) in the male. (C) Cloacal region of

female A. ansorgii, FKH0009. Note lack of precloacal pores on

the chevron row of enlarged scales or of enlarged tubercular

lateral scales on tail base. Both sexes have delicate elongate

toes with a single pair of terminal scansors. The cylindrical tail

is finely scaled, and non-verticillate, lacking obvious lateral

constrictions.

August 2019 | Volume 13 | Number 2 | e182

Rediscovery of Afrogecko ansorgii in Angola

advective fogs which become trapped among high rocky

slopes. Although lichens were less abundant at Maconjo,

the type locality also appeared to be regularly subject

to thick foggy conditions. Situated at around 40 km

from the coast and 400 m asl, Maconjo lies at the base

of a north-south oriented orographic step, characterized

by mountainous granitic hills that elevate the coastal

plateau further east to above 800 m asl, and therefore

topography likely plays a role in containing fog and

low strata clouds and, ultimately, shaping the ecological

conditions of local spiny savannas. The blackthorn bush

holds a reputation for being termite-resistant (Schmidt et

al. 2002). Nevertheless, in both Maconjo and Sta Maria

we found ‘drywood’ termites (family Kalotermitidae) to

be present inside dead blackthorn branches, often leaving

them neatly excavated. This was especially evident at the

drier environment of Sta Maria, where large numbers

of dead branches break and accumulate on the ground

underneath the bush canopy, thus providing an abundance

of sheltering habitat for the gecko. Drywood termites are

well adapted to arid ecosystems but may also require

some moisture and tend to be uncommon or patchily

distributed. It appears therefore that the local distribution

of termites is driven by specific environmental conditions

affecting the bushes of Senegalia mellifera detinens, like

altitude and rainfall, atmospheric moisture provided

by fog or low clouds, and the frequency and severity

of droughts. In regions with relatively higher moisture

from average rainfall, these blackthorn savannas may not

accumulate enough dead wood and the conditions might

be unsuitable for the termites and ultimately the gecko.

On the other hand, under extremely dry conditions the

termites may not thrive, or the density of bushes might be

too low, not providing enough shelter or foraging habitat

for this exclusively arboreal gecko. Therefore, the dry

coastal blackthorn savannas that experience low rainfall

but are subject to frequent fogs, seem to constitute the

‘sweet spot’ habitat for the species.

Formerly known locally as Deserto de Mossamedes,

the northernmost extension of the Namib Desert stretches

north along the Atlantic coast of Angola from the Angola-

Namibia border for 450 km to about the city of Benguela.

Fronting the Atlantic Ocean to the west, it gradually

ascends in elevation eastward to a semiarid plain

dominated by acacia and mopane [African ironwood]

trees that abuts the steep Serra de Chela escarpment.

Characterized by gravel plains and rock platforms

interspersed with sand dune fields, the Angolan Namib

is part of a broader ecoregion defined as Kaokoveld

Desert (Burgess 2004). It is believed that ancient climatic

stability, a mosaic of substrates and incidence of coastal

fogs contribute to high species richness and rates of

endemism in the Namib region (Seeley et al. 1998), and

the Kaokoveld is often referred to as a regional center

of endemism for flora (Van Wyk and Smith 2011),

beetles (Koch 1961), and lizards (Lewin et al. 2016;

Branch et al. 2019b). Nevertheless, the boundaries of the

northern Kaokoveld have remained poorly defined and

although Burgess (2004) depicted the ecoregion along

the Angolan coast to about Benguela town, other authors

have set the northern limit at Lucira, in Namibe Province

(e.g., Craven 2009; Branch et al. 2019). Between Lucira

Amphib. Reptile Conserv.

and Benguela the semi-arid ecosystems characterized by

a diversity of succulent plants progressively gives way

to semi-arid dry savannas dominated by acacia species

(Barbosa 1970), thus placing our surveyed sites on the

fringes of the Kaokoveld ecoregion. Historical and recent

herpetological work on the Angolan arid ecosystems

has mostly focused in the southernmost regions of the

Angolan Kaokoveld (Marques et al. 2018; Branch

et al. 2019b), and about one-third of national reptile

diversity is known to occur in the Namibe Province

alone, with gekkonids being the most speciose group

(Ceriaco et al. 2016). In comparison, less effort has

been directed to Benguela Province, yet the diversity

and new extension ranges here reported suggest that

the herpetological richness of the province may have

been underestimated. It is likely that future surveys

across the northern limits of the Kaokoveld will further

increase the regional lists, unveil cryptic diversity, and

underline the ecological significance of the local spiny

semi-arid savannas associated with fog and low strata

clouds. Apparently restricted to Benguela’s Kaokoveld

and strongly associated with the local fog ecosystem,

the now rediscovered Ansorge’s gecko remains as one of

the most unique and iconic representatives of Angolan

herpetofauna.

Acknowledgements.—We thank our various companions

for their help and camaraderie, both 1n the field and in

general: Ninda Baptista, Afonso Vaz Pinto, Kostadin

Luchansky, and Werner Conradie. Scientific collaboration

is enriched by such synergy and stimulation. In addition,

we thank the National Geographic Okavango Wilderness

Project (National Geographic Society grant number

EC0715-15) for funding fieldwork and biodiversity

surveys in Angola, and the Ministry of Environment

(MINAMB) for issuing research and export permits. We

thank Wulf Haacke for discussion on his early exploration

of Angola in the 1970s, when he made numerous

discoveries, many still unpublished. We are also indebted

to Fernanda Lages and to ISCED — Jnstituto Superior de

Ciéncias da Educagao da Huila for logistical support.

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August 2019 | Volume 13 | Number 2 | e182

Vaz Pinto et al.

Pedro Vaz Pinto is Angolan and was born 1n Luanda, Angola in 1967. Pedro graduated

in Forest Engineering at Technical University of Lisbon, and obtained a doctoral

degree in Biology from the University of Porto, Portugal. Over the past 20 years,

he has worked in biodiversity conservation in Angola addressing rare or endangered

species, and protected area management. Pedro is a director for the local NGO Kissama

Foundation, and a researcher for CIBIO-InBio. His studies on Angolan vertebrates

have focused mostly on genetics, biogeography, and conservation in antelopes, birds,

reptiles, and amphibians. Pedro travels the country extensively and has received three

international environmental awards for his biodiversity conservation work in Angola.

Luis Verissimo is Portuguese and was born in Lisbon, Portugal, in 1971. He graduated

in Geography from the University of Lisbon, and obtained a Master’s degree in Applied

Ecology from the Michigan Technological University in the United States. Luis is a

geospatial specialist and ecologist with 20 years of experience conducting geospatial

data analysis, cartography, and field and survey investigations for land-based,

freshwater, and marine applications. He has worked extensively on the vertebrates of

Angola addressing biogeography, conservation, and historical distribution. Luis has

also been actively engaged in the assessment of existing and establishment of new

protected areas within Angola’s framework of national parks and reserves.

Bill Branch (William R. Branch) was born in London, United Kingdom. He was employed as Curator of Herpetology at

the Port Elizabeth Museum for over 30 years (1979-2011), and upon his retirement he was appointed Curator Emeritus

Herpetology until his death in October 2018. Bill’s herpetological studies concentrated mainly on the systematics,

phylogenetic relationships, and conservation of African reptiles, but he has been involved in numerous other studies on

the reproduction and diet of African snakes. He has published over 300 scientific articles, as well as numerous popular

articles and books. The latter include: South African Red Data Book of Reptiles and Amphibians (1988), Dangerous

Snakes of Africa (1995, with Steve Spawls), Field Guide to the Reptiles of Southern Africa (1998), Tortoises, Terrapins

and Turtles of Africa (2008), and Atlas and Red Data Book of the Reptiles of South Africa, Lesotho and Swaziland

(multi-authored, 2014), as well as smaller photographic guides. In 2004, he was the 4" recipient of the “Exceptional

Contribution to Herpetology” award of the Herpetological Association of Africa. Bill has undertaken field work in over

16 African countries, and described nearly 50 species, including geckos, lacertids, chameleons, cordylids, tortoises,

adders, and frogs.

Amphib. Reptile Conserv.

41 August 2019 | Volume 13 | Number 2 | e182

Official journal website:

amphibian-reptile-conservation.org

Amphibian & Reptile Conservation

13(2) [Special Section]: 42—56 (e184).

€ptile-cons™

Phylogeography of the East African Serrated Hinged Terrapin

Pelusios sinuatus (Smith, 1838) and resurrection

of Sternothaerus bottegi Boulenger, 1895 as a subspecies

of P. sinuatus

‘Melita Vamberger, 7Margaretha D. Hofmeyr, *Courtney A. Cook,

45Edward C. Netherlands, and °*Uwe Fritz

‘6 Museum of Zoology, Senckenberg Natural History Collections Dresden, A. B. Meyer Building, 01109 Dresden, GERMANY ?Chelonian Biodiversity

and Conservation, Department of Biodiversity and Conservation Biology, University of the Western Cape, Bellville 7535, SOUTH AFRICA ?4Unit

for Environmental Sciences and Management, North-West University, Private Bag X6001, Potchefstroom 2520, SOUTH AFRICA *Laboratory of

Aquatic Ecology, Evolution and Conservation, KU Leuven, Charles Deberiotstraat 32, 3000 Leuven, BELGIUM

Abstract.—Pelusios sinuatus is distributed in East Africa from southern Ethiopia and Somalia to northeastern

South Africa. Inland it reaches westernmost Zimbabwe, Rwanda, and Burundi. Despite this wide range, which

spans in north-south direction across 3,500 km and in east-west direction more than 1,500 km, no geographic

variation has been described. However, using phylogenetic and haplotype network analyses of mitochondrial

and nuclear DNA (2,180 bp and 2,132 bp, respectively), phylogeographic variation is herein described, with two

distinct genealogical lineages. One occurs in the northern and central parts of the distribution range, and the

other is in the south. Terrapins representing the southern lineage attain a smaller maximum body size than

terrapins from the northern and central parts of the range. The distribution ranges of the two lineages abut

in the border region of Botswana, South Africa, and Zimbabwe. We conclude that each lineage represents a

distinct subspecies, with the nominotypical subspecies Pelusios sinuatus sinuatus (Smith, 1838) occurring in

the south and the newly recognized subspecies Pelusios sinuatus bottegi (Boulenger, 1895) in the central and

northern distribution range. We found phylogeographic structuring within each subspecies and propose that

the differentiated population clusters should be recognized as Management Units.

Keywords. management units, Pelomedusidae, systematics, taxonomy, Testudines, turtle

Citation: Vamberger M, Hofmeyr MD, Cook CA, Netherlands EC, Fritz U. 2019. Phylogeography of the East African Serrated Hinged Terrapin

Pelusios sinuatus (Smith, 1838) and resurrection of Sternothaerus bottegi Boulenger, 1895 as a subspecies of P. sinuatus. Amphibian & Reptile

Conservation 13(2): [Special Section]: 42-56 (e184).

tion 4.0 International (CC BY 4.0): https://creativecommons.org/licenses/by/4.0/], which permits unrestricted use, distribution, and reproduction in any

medium, provided the original author and source are credited. The official and authorized publication credit sources, which will be duly enforced, are

as follows: official journal title Amphibian & Reptile Conservation; official journal website: amphibian-reptile-conservation.org.

Received: 6 February 2019; Accepted: 5 June 2019; Published: 19 August 2019

Introduction Pelomedusids and podocnemidids are both of Gondwanan

origin (de Broin 1988; Noonan 2000) and, together with

The freshwater turtle genus Pe/usios comprises 17 — the Australasian and South American family Chelidae,

species distributed across sub-Saharan Africa, with — represent the chelonian suborder Pleurodira (side-necked

most likely introduced populations on Madagascar, the turtles; TTWG 2017).

Seychelles, and Guadeloupe in the Lesser Antilles (Fritz In addition to a number of species with localized

et al. 2011, 2013; Stuckas et al. 2013; TTWG 2017). All narrow distribution, Pe/usios also includes species

species of Pe/usios are side-necked turtles characterized — with wide distributions (Branch 2008; TTWG 2017).

by a plastral hinge that allows partial or complete closure The Serrated Hinged Terrapin, P sinuatus (Smith,

of the anterior shell opening (Bramble and Hutchison 1838), has one of the largest distribution ranges of all

1981; Branch 2008). Together, Pe/usios and its African- — Pe/usios species. It occurs in East Africa from southern

Arabian sister genus Pelomedusa, which has 10 formally — Ethiopia and Somalia southwards to Eswatini (formerly

recognized species (Petzold et al. 2014; TTWG 2017), Swaziland) and northeastern South Africa (Branch 2008;

constitute the family Pelomedusidae that is sister to ©TTWG 2017; Fig. 1). The Serrated Hinged Terrapin has

the New World-Madagascan family Podocnemididae. _ the largest body size in its genus and may reach up to

Correspondence. * wwe.fritz@senckenberg.de

Amphib. Reptile Conserv. 42 August 2019 | Volume 13 | Number 2 | e184

Vamberger et al.

Botswana

Mozambique A3

Botswana

South Africa

(Limpopo, B1

Mpumalanga)

Pelusios sinuatus 1.0

Mozambique

Zimbabwe

South Africa

(KwaZulu-Natal, B2

Limpopo)

Pelusios marani

0.4

Fig. 1. Left: Sampling sites and mitochondrial identity of Pe/usios sinuatus used in the present study. Divided symbols indicate

syntopic occurrences of the respective subclades. Terrapin shapes symbolize differences in maximum sizes of the northern and southern

P. sinuatus (see Discussion section). Inset: Distribution range of P. sinuatus according to the TTWG (2017) with the type localities

of taxa referred to this species. (1) Sternothaerus bottegi Boulenger, 1895: Bardere (Bardera), Somalia; (2) Pelusios sinuatus leptus

Hewitt, 1933: Isoka, Zambia; (3) Sternotherus sinuatus Smith, 1838 — restricted type locality (Broadley, 1981): confluence of Crocodile

and Marico Rivers, Limpopo (Transvaal); (4) Pe/usios sinuatus zuluensis Hewitt, 1927: Mzinene River (Umsinene River, Zululand),

KwaZulu-Natal. Right: Bayesian tree for 61 Pe/usios sinuatus using 2,180 bp of mitochondrial DNA. Clades are collapsed to cartoons

showing the deepest genetic divergence within each clade. Outgroup Pelomedusa variabilis was removed for clarity. Numbers above

the nodes are posterior probabilities; below the nodes, thorough bootstrap values under ML. Full trees are available from https://

figshare.com/s/52a7af23cff3aa08ea75. Inset: Adult Pelusios sinuatus, Bonamanzi, KwaZulu-Natal, South Africa.

55 cm in shell length (Spawls et al. 2002). In contrast a distribution range similar to that of P sinuatus, was

to other Pelusios species, P. sinuatus is a deep-water — characterized by moderate phylogeographic variation

terrapin that occurs in perennial rivers, lakes, and larger _— (Fritz et al. 2013).

man-made water bodies in savannah regions. During the Until now, neither the morphological nor the

rainy season, Serrated Hinged Terrapins move overland, phylogeographic variation of P sinuatus has been

and they colonize smaller water bodies, like pans and studied systematically. The species has traditionally

waterholes (Broadley and Boycott 2009). In eight other been regarded as monotypic (Ernst and Barbour 1989;

widespread Pelusios species, Fritz et al. (2013) and TTWG 2017; Wermuth and Mertens 1961, 1977),

Kindler et al. (2016) found that phylogeographic patterns = even though Hewitt (1927, 1933) had described two

were not correlated with habitat type, with some species — subspecies from South Africa and Zambia that were

displaying pronounced phylogeographic structuring soon synonymized (Loveridge 1936). However, using

and others not. Among the studied savannah species, P. mitochondrial and nuclear DNA sequences of only two

rhodesianus showed a deep phylogeographic structure samples from KwaZulu-Natal (South Africa) and another

and could actually represent a species complex (Kindler — one from Botswana, Fritz et al. (2011) found two genetic

et al. 2016), whereas phylogeographic structuring in lineages, suggesting that an in-depth investigation of

P. nanus and P. subniger was negligible. However, the — genetic variation may reveal further differentiation.

westernmost studied population originally identified as § The present paper presents the first assessment of the

P. subniger (in Democratic Republic of the Congo) was genetic variation of P sinuatus across its range. The

found to represent a genetically distinct undescribed results are discussed with respect to taxonomy and two

species. Unfortunately, samples from the northern — subspecies are recognized within P. sinuatus. To this end,

distribution area of P. subniger were not available for | Sternothaerus bottegi Boulenger, 1895 is resurrected

study, so nothing is known about the genetic identity of | from the synonymy of P. sinuatus (Smith, 1838), in

the northern populations (Fritz et al. 2013; Kindler et al. | which it was placed soon after its description (Calabresi

2016). Another savannah species (P. castanoides), with 1916; Siebenrock 1916).

Amphib. Reptile Conserv. 43 August 2019 | Volume 13 | Number 2 | e184

Phylogeography of Pelusios sinuatus in East Africa

Materials and Methods

Sampling, chosen loci, and general data evaluation

strategy: Sixty-one samples of Pe/usios sinuatus from

Botswana, Mozambique, South Africa, and Tanzania

were studied, including previously published data for

three terrapins (Appendix 1). The same mitochondrial

and nuclear DNA fragments were targeted as in earlier

studies on Pelusios (Fritz et al. 2011; Kindler et al. 2016).

Three mitochondrial DNA fragments were sequenced

(12S, cyt 6, and ND4 with adjacent DNA coding for

tRNAs). In addition, two protein-coding nuclear genes

(Cmos and Rag2) and intron 1 of the nuclear R35 gene

were sequenced. Details of DNA isolation, PCR, and

sequencing are described in Kindler et al. (2016). The

12S sequences obtained were up to 398 bp long (with

gaps); the cyt b sequences were up to 913 bp; and the

mtDNA sequences comprising the partial ND4 gene

plus adjacent DNA coding for tRNAs were up to 869

bp long. All nuclear DNA blocks could be sequenced

directly. Cmos sequences had lengths of up to 358 bp;

R35 sequences, up to 1,101 bp; and Rag2 sequences,

up to 673 bp. Sequences were aligned and inspected

using BioEdit 7.0.5.2 (Hall 1999). All sequences aligned

perfectly and gaps occurred only in sequence blocks not

coding for proteins.

Mitochondrial DNA is maternally inherited, whereas

nuclear loci are inherited biparentally. Moreover,

mtDNA is prone to introgression, including across

species borders, which often leads to conflicting results

for the two marker systems (Currat et al. 2009; Funk and

Omland 2003; Kehlmaier et al. 2019; Sloan et al. 2017;

Toews and Brelsford 2012). To avoid the risk of such

distortion, mitochondrial and nuclear sequence data were

examined separately.

Phylogenetic analyses: Individual mtDNA fragments

were concatenated for phylogenetic analyses, and this data

set was combined with previously published sequences,

resulting in an alignment of 2,180 bp length. The dataset

included 61 sequences of Pelusios sinuatus and, as

outgroups, one sequence each of Pelomedusa variabilis

and Pelusios marani. European Nucleotide Archive

(ENA) accession numbers and collection sites are given in

Appendix |. The best partitioning scheme was determined

using PartitionFinder (Lanfear et al. 2012) and the

Bayesian Information Criterion (BIC). Three partitioning

schemes were tested: (1) unpartitioned, (2) partitioned by

mtDNA fragment, and (3) partitioned by gene and codon

position with DNA not coding for proteins (1.¢e., 12S and

DNA coding for tRNAs) corresponding to one additional

partition each. According to the results of PartitionFinder,

scheme (3) was selected.

Phylogenetic relationships were inferred using

Bayesian and Maximum Likelihood (ML) approaches.

Bayesian trees were obtained with MrBayes 3.2.6

(Ronquist et al. 2012) using the partitioning scheme

and evolutionary models shown in Table 1 and default

parameters. Two parallel runs, each with four chains, were

conducted. The chains ran for 10 million generations,

with every 500" generation sampled. The calculation

parameters were analyzed using a burn-in of 2.5

Amphib. Reptile Conserv.

million generations to assure that both runs converged.

Subsequently, only the plateau of the remaining trees

was sampled using the same burn-in, and a 50% majority

rule consensus tree was generated. Tracer 1.7 (Rambaut

et al. 2018) served to check for convergence of the runs

using the Effective Sample Sizes (ESS) of parameters,

and resulted in ESSs over 200 after discarding the burn-

in. In addition, phylogenetic relationships were inferred

under ML using RAxML 7.2.8 (Stamatakis 2006) and

the GTR+G substitution model across all partitions. Five

independent ML searches were performed using different

starting conditions and the fast bootstrap algorithm

to explore the robustness of the results by comparing

the best trees. Then, 1,000 non-parametric thorough

bootstrap replicates were calculated and the values were

plotted against the best tree.

Parsimony networks: For each mitochondrial and

nuclear DNA fragment, a parsimony network was

constructed using Popart (http://popart.otago.ac.nz).

Since the underlying TCS algorithm is sensitive to

missing data, a few individuals represented by short

sequences were excluded. In addition, for achieving

complete coverage, the lengths of mtDNA sequences

were trimmed, resulting in an alignment of 348 bp length

for 12S, 784 bp for cyt 5, and 737 bp for ND4 + DNA

coding for tRNA. For network construction of nuclear

data, heterozygous sequences of R35 were phased

using the Phase algorithm in DnaSP 5.10 (Librado and

Rozas 2009), and two identical copies for homozygous

sequences of all loci were included. Nuclear DNA

sequences for the networks had the same lengths as given

above.

Uncorrected p distances and isolation by distance for

mtDNA: Uncorrected p distances were calculated for the

mitochondrial cyt b gene alone as well as for the mtDNA

alignment of concatenated sequences using MEGA

7.0.21 (Kumar et al. 2016) and the pairwise deletion

option. The distances of the concatenated sequence data

were used to examine for a positive correlation between

geographic and genetic distances (isolation by distance).

For this purpose, Mantel tests as implemented in IBD

1.52 (Bohonak 2002) were run for three data sets using

genetic and spatial distances. The latter were obtained via

the Geographic Distance Matrix Generator 1.2.3 (http://

biodiversityinformatics.amnh.org/open_source/gdmg/

index.php). The significance of the slope of the reduced

major axis (RMA) regression was assessed by 30,000

randomizations. One data set included the sequences

for all 61 terrapins. The other two data sets included the

sequences for each clade of P. sinuatus (clades A and B)

identified in the present study.

Table 1. Partitioning and evolutionary models used for

MrBayes.

Subset nst rates Model

1-398 401-1311\3 1314- 6 gamma SYM+ gamma

1998\3 1999-2180

399-1311\3 1312-1998\3 6 SYM

400-1311\3 1313-1998\3 6 gamma SYM+ gamma

August 2019 | Volume 13 | Number 2 | e184

Vamberger et al.

Body size: Using Vernier calipers (accuracy 0.1 mm),

straight carapace length of adult terrapins was recorded as

a measure of body size during fieldwork in South Africa

(n = 11). Also measured were specimens in the Museum

for Comparative Zoology, Cambridge, Massachusetts,

USA (MCZ 39383, South Africa), the Museum fir

Naturkunde, Berlin, Germany (ZMB 158, Mozambique;

ZMB 5517, 5518, 15689, 16158, 16242, Tanzania; ZMB

22416, Burundi), the Museum fiir Tierkunde (Museum

of Zoology), Senckenberg, Dresden, Germany (MTD D

49650, South Africa), and the Palaontologisches Institut

und Museum, Zurich, Switzerland (PIMUZ A/III527,

Tanzania). Measurements were divided into northern

and southern groups according to the two genetic clades

identified in the present study. Since males and females

could not be distinguished in all samples, the two sexes

were combined for analysis. After testing for whether the

data were parametric, the body sizes of the groups were

compared using a f-test as implemented in SigmaPlot

13.0 (Systat Software, Inc., San Jose, California, USA).

Results

Phylogenetic and haplotype network analyses of

mtDNA: Both tree-building approaches delivered the

same topology, corresponding to two geographically

widespread clades, A and B, both of which showed

substructuring and received high support values (Fig.

1). Clade A corresponded to the samples from the north

and center of the distribution range of Pe/usios sinuatus

(Tanzania, Mozambique, and Botswana), and clade B to

samples from the south (Botswana and South Africa).

Clade A consisted of three subclades and clade B, of

two. All subclades of clade A and one subclade of clade

B were well supported under both Bayesian and ML

analyses; the second subclade of clade B was moderately

supported. In Botswana and northeastern South Africa,

records of terrapins representing clades A and B are only

separated by a distance of approximately 200 km, and in

northeastern South Africa, representatives of subclades

B1 and B2 were found in two sites syntopically.

In haplotype networks of the three mtDNA fragments,

no shared haplotypes occurred for the two clades (Fig. 2).

Four 12S haplotypes were found for clade A, with two

private haplotypes for subclade A2 that differed by one

mutation step each from a shared haplotype that included

sequences of subclades Al and A3. Another private

haplotype of subclade A3 also differed by one mutation

step from the previously mentioned shared haplotype.

This shared haplotype was separated by three mutation

steps from a common haplotype containing sequences

of subclades B1 and B2, and a second rare haplotype

for subclade B1 differed by one mutation step from this

common haplotype. Haplotype networks of the two other

mtDNA fragments showed more differentiation, with

no shared haplotypes between any clades or subclades.

For the cyt 5 fragment, haplotypes of clades A and

B were separated by a minimum of 21 steps. Within

haplotypes of clade A, up to 10 mutations occurred, with

each subclade corresponding to one distinct haplotype.

Within haplotypes of clade B, a loop occurred that

connected three of the four haplotypes of subclade B1;

Amphib. Reptile Conserv.

and the four haplotypes of this subclade were separated

by a minimum of four steps from the three haplotypes

of subclade B2, each of which differed by one mutation

step. With respect to the mtDNA fragment containing the

partial ND4 gene and adjacent DNA coding for tRNAs,

haplotypes of clades A and B differed by a minimum

of 13 mutations. Subclade Al was represented by one

haplotype. Subclade A2 consisted of three haplotypes

that each differed by a maximum of three mutations; and

subclade A3 had four haplotypes that differed by up to

four steps. Within the individual haplotypes of clade A, a

maximum of nine steps occurred, and the three subclades

were separated by a minimum of 4—7 steps. The three

haplotypes of subclade B1 differed by a maximum of

two mutations and the two haplotypes of subclade B2,

by one mutation, and the two subclades were distinct by

a minimum of six steps.

Haplotype network analyses of nuclear loci: The three

nuclear loci showed distinctly less variation compared to

the mtDNA fragments. Often, shared haplotypes between

distinct clades and subclades were found (Fig. 3). For

the Cmos gene, the 15 haplotypes found differed by a

maximum of nine mutation steps. Clades A and B shared

two haplotypes, even though the vast majority of phased

sequences of clades A and B corresponded to unique

haplotypes for each clade. A generally similar picture was

revealed for the Rag2 locus, with one shared haplotype of

clade A and clade B, six unique additional haplotypes of

clade A, and three further unique haplotypes of clade B.

The maximum number of mutations between the Rag2

haplotypes was seven. For intron 1 of the R35 gene, no

shared haplotypes were found for clades A and B. A total of

15 haplotypes occurred that were partially connected over

a loop. In a direct line (not across the loop), the haplotypes

differed by up to seven mutations. Of the 15 haplotypes,

six corresponded to clade A and nine to clade B.

Uncorrected p distances and isolation by distance for

mtDNA: Sequence divergences of the mitochondrial cyt

b gene are often used to distinguish between chelonian

taxa (e.g., Iverson et al. 2013; Kindler et al. 2012,

2016; Petzold et al. 2014). The uncorrected p distance

between clade A and clade B of P. sinuatus amounted to

2.80% on average, while the within-group values were

1.05% and 0.28%, respectively. Between the individual

subclades of clade A, divergences ranged between 1.31%

and 1.54%, with no variation within those two clades

for which sequences of more than one individual were

available. Subclades B1 and B2 differed by only 0.57%,

with within-group divergences of 0.06% and 0.02%,

respectively (Table 2).

The IBD test for all data revealed a statistically

significant correlation of genetic and geographic distances

(Z = 22487528, r = 0.65, p < 0.0001; n = 61). When

the two clades were analyzed separately, a statistically

significant correlation was also found for clade A (Z =

1387859, r= 0.58, p < 0.0003; n= 17), but not for clade

B (Z = 9009469796357, r = 0.06, p < 0.0777; n= 44).

Body size: The mean straight carapace length (+ SD)

for the samples of the northern and southern clades,

August 2019 | Volume 13 | Number 2 | e184

Phylogeography of Pe/usios sinuatus in East Africa

ies @ ai

3 12S a es @ A2

n=60 7 » © A3

©) B1

18 2

cyt b

n=48 3

ND4+tRNAs

n=61

Fig. 2. Parsimony networks for individual mtDNA fragments of Pelusios sinuatus. Symbol size is proportional to haplotype

frequency. Each line connecting two haplotypes corresponds to one mutation step, if not otherwise indicated by numbers of

substitutions along the lines. Colors correspond to Fig. 1. Small black circles represent missing node haplotypes.

n=106 ac

Fig. 3. Parsimony networks for nuclear loci of Pe/usios sinuatus. Symbol size is proportional to haplotype frequency. Each line

connecting two haplotypes corresponds to one mutation step, if not otherwise indicated by numbers of substitutions along the lines.

Colors correspond to Fig. 1. The small black circle represents a missing node haplotype. Sample sizes refer to phased nuclear

sequences, i.e., each individual is represented twice.

Amphib. Reptile Conserv. 46 August 2019 | Volume 13 | Number 2 | e184

Vamberger et al.

HB North n=7

South n=14

Individuals

| | } I

0 -

30 35 40 45

a 10 15 20 25

Carapace length (straight line, cm)

Fig. 4. Carapace lengths of adult Pel/usios sinuatus from

the northern and the southern distribution range (museum

specimens and wild-caught terrapins), corresponding to clades

A (north) and B (south).

respectively, were 31.8 + 7.9 cm (n= 7) and 24.0 + 5.5

cm (n= 14), with terrapins from the northern clade being

significantly larger than those from the southern clade (t,,

= 2.66, p = 0.0156).

Discussion

The present study is the first assessment of the

phylogeography for the Serrated Hinged Terrapin

(Pelusios sinuatus), which is widely distributed in

East Africa (TTWG 2017; inset in Fig. 1). In north-

south direction, the distribution area extends over

approximately 3,500 km and in east-west direction,

over more than 1,500 km. Within this large area,

two mitochondrial clades (A and B) with parapatric

distribution and substantial geographic substructure were

discovered (Figs. 1 and 2). In contrast to mitochondrial

DNA, the slower evolving nuclear DNA has not reached

complete lineage sorting for the Cmos and Rag2 loci

(Fig. 3), even though haplotype sharing between clades

A and B was restricted. For intron 1 of the R35 gene,

no shared haplotypes occurred. Thus, mitochondrial and

nuclear markers show largely concordant differentiation

patterns.

Clade A was found in the northern and central parts

of the distribution range (Tanzania, Mozambique, and

Botswana), and clade B, in the south (Botswana and

South Africa). Close to the border region of Botswana,

Zimbabwe, and South Africa the two clades abut, which

explains why Fritz et al. (2011) had already discovered

the two clades using only three samples from that

region. Nearby, to the southeast, sites were found with

syntopic occurrences of the two otherwise parapatric

mitochondrial subclades within clade B. This implies

that the correlation of genetic and geographic distances

for the whole data set cannot result from isolation by

distance alone, because then neither distinct clades nor

contact zones would be expected (Figs. 1 and 2). This is

also supported by the absence of evidence for isolation by

distance in the southern clade B. Therefore, we conclude

Amphib. Reptile Conserv.

Table 2. Uncorrected p distances (means, expressed as

percentages) between and within mitochondrial subclades of

Pelusios sinuatus using the cyt b gene (913 bp). Below the

diagonal are between-group values; on the diagonal, within-

group divergences are in bold.

n Al A2 A3 Bl B2

Al 2 0

A2 1 1.31 —

A3 3 Is53; 1.31 0

Bl 1B) 2.83 2.85 263 0.06

B2 21 2:15 297, 273 0.57 0.02

that the observed genetic divergence is, at least in part,

caused by vicariance and subsequent dispersal, and

that the correlation of geographic and genetic distances

results mainly from our patchy sampling. A future

challenge is to close the large sampling gaps in order to

locate additional contact zones, especially between the

northern subclades.

The north-south differentiation of P. sinuatus is similar

to that in another terrapin species. Pelusios castanoides

has a continental distribution range similar to P. sinuatus

but occurs also on Madagascar and the Seychelles

(TTWG 2017), although it is unclear whether the latter

populations are native. In P. castanoides, a sample

from the north of the distribution range (Kenya) was

distinct from samples from South Africa and southern

Mozambique (Fritz et al. 2013), suggesting a shared

biogeographic history with P. sinuatus.

To the best of our knowledge, P. sinuatus from

different parts of the distribution range have never

been compared morphologically, but northern terrapins

(clade A) seem to grow to a larger maximum size than

southern ones (clade B). According to de Witte (1952),

P. sinuatus reaches 46.5 cm in Lake Tanganyika. Branch

(2008) reports a maximum shell length of 48.5 cm for

P. sinuatus, and Spawls et al. (2002) mention up to 55

cm for upland Kenya. Serrated Hinged Terrapins of such

size are never seen in South Africa. This is confirmed

by the measurements of wild animals and collection

material reported here (Fig. 4) that show a statistically

significant difference between the mean carapace lengths

of terrapins representing clades A and B. Seven adult

museum specimens from Tanzania and Burundi (clade

A) had straight carapace lengths from 19.6 cm to 40.0 cm

(i.e., the largest specimens are still distinctly below the

published maximum size). In contrast, 14 adult museum

specimens and wild terrapins from the distribution range of

clade B (Mozambique and South Africa) ranged between

17.5 cm and 34.7 cm. Thus, it appears that northern and

southern terrapins differ morphologically, at least with

respect to their maximum size, but more measurements

are required and further studies warranted for comparing

variations in additional morphological characters. The

size variation of P. sinuatus is reminiscent of other turtles

in which size increases with latitude (Ashton and Feldman

2003), either within the same species (e.g., Chelonoidis

chilensis: Fritz et al. 2012a; Testudo graeca: Werner et al.

2016) or in distinct species of the same genus (Pelodiscus

spp.: Farkas et al. 2019). However, there are exceptions.

For instance, in Leopard Tortoises (Stigmochelys

August 2019 | Volume 13 | Number 2 | e184

Phylogeography of Pelusios sinuatus in East Africa

Table 3. Average uncorrected p distances (percentages) of 795 bp of the cyt b gene of Pe/usios species from Kindler et al. (2016).

Pelusios subniger includes a putative undescribed species from the Democratic Republic of the Congo (n = 2). It differs from other

P. subniger by an average distance of 3.13% (Petzold et al. 2014). The relationship of P. carinatus and P. rhodesianus is unclear;

some populations assigned to the latter species (P. rhodesianus A) could be conspecific with P. carinatus (Kindler et al. 2016).

Values for sympatrically occurring species pairs in boldface and red.

nig rhoA rhoB

sin

sub

upe

wil

10.91 14.01 11.82 14.99 13.56 14.56 13.54 11.80 5.43

n ada bec bro car c‘us c‘es cha cup gab mar nan

adansonii

bechuanicus 2; JLINS9

broadleyi TE e305 “1315

carinatus 15. 9:92 12.91 11.17

castaneus 19 652 13.70 7.55 11.48

castanoides 29 9.57 11.50 9.88 10.99 11.17

chapini 8 6.03 15.91 10.17 13.95 4.34 12.35

cupulatta 4 10.98 10.79 11.24 11.26 12.14 10.76 15.03

gabonensis 24 12.73 13.62 13.68 12.84 13.52 12.54 12.42 13.09

marani 5 12.80 11.21 12.68 13.01 13.57 12.50 15.35 11.82 13.75

nanus 26 11.59 14.99 15.07 15.31 15.36 12.21 12.94 14.09 12.42 15.25

niger 11.76 13.38 13.90 12.25 15.17 11.85 15.77 9.10 12.73 14.14 13.07

rhodesianus A 10.81 13.45 11.39 2.49 11.83 11.12 10.92 12.28 11.43 14.19 13.24 12.57

rhodesianusB 14 7.93 14.23 11.93 6.22 12.39 12.12 10.63 13.44 12.09 14.58 12.41 12.77 4.04

sinuatus 2 14.02 11.42 13.05 12.52 14.01 13.60 16.32 11.26 13.95 11.02 15.73 14.34 13.92 13.96

subniger 41 14.57 5.33 12.25 12.70 13.50 11.89 15.28 10.92 13.15 11.21 13.72 11.64 13.50 13.55 10.08

upembae 3 10.98 1.38 13.15 13.04 14.41 11.50 16.54

williamsi 2 915 10.91 10.87 10.23 11.03 3.89 15.00

pardalis) the northernmost and southernmost populations

comprise large-sized individuals, while tortoises from

geographically intermediate populations are medium-

sized (Fritz et al. 2010; Spitzweg et al. 2019). Another

pattern is found in continental Trachemys species once

considered conspecific, with taxa having the largest

body sizes in Central America and distinctly smaller-

sized North and South American congeners (Ernst and

Barbour 1989; Legler and Vogt 2013; Vargas-Ramirez

et al. 2017). Clearly, further research is needed for a

better understanding of the described size variation in P.

sinuatus and other turtle species, but we concur with Joos

et al. (2017) and Spitzweg et al. (2019) that many factors

beyond latitude act in concert on such variation.

An open question remains how the genetic and

morphological differentiation patterns of P. sinuatus relate

to taxonomy. The concordant variation of different genetic

and morphological characters justifies recognizing each

clade within P. sinuatus as a distinct taxon. However,

without entering the debate about species concepts and

Species conceptualization (e.g., de Queiroz 2007; Zachos

2016), we are reluctant to assign species status to either

clade. In our understanding, restricted gene flow and

largely isolated gene pools represent unambiguous

traits of distinct species. In contrast to other cases (e.g.,

Kindler et al. 2017; Spinks et al. 2014; Vamberger et al.

2015), patchy sampling prevents sound conclusions here,

particularly the lack of comprehensive sampling from the

putative contact zone of clades A and B. Yet, in times when

legislative restrictions make biodiversity research virtually

impossible for many widely distributed species (Neumann

et al. 2018; Prathapan et al. 2018), researchers are often

forced to use the evidence available as a starting point.

Amphib. Reptile Conserv.

11.01 14.11 11.33 15.73 13.69 13.97 10.77 13.31 12.28 11.31

The mitochondrial divergence of the two clades (Figs.

1 and 2), together with concordant variation in the nuclear

loci (Fig. 3), provide two important insights: (1) the two

mitochondrial clades represent distinct genealogical

lineages; and (2) mitochondrial introgression plays no

obvious role here, allowing the application of mtDNA to

infer taxonomic differentiation. Uncorrected p distances

of the mitochondrial cyt b gene have frequently been used

as a ‘taxonomic yardstick’ to decide which taxonomic

rank should be applied to turtle taxa (e.g., Iverson et al.

2013; Kindler etal. 2012, 2016; Petzold et al. 2014; Spinks

et al. 2004), analogous to the widely applied barcoding

approach (e.g., Hebert et al. 2003). However, as pointed

out by Fritz et al. (2012b) and Kindler et al. (2012), the

wide range of genetic divergences between different

turtle species (differing by one order of magnitude)

prevents the application of a rigid threshold across all

turtle groups. Instead, thresholds for different groups

need to be adjusted individually using unambiguous,

ideally sympatric, species that are closely related to the

taxa in question. Thus, previously published cyt b data

for other Pe/usios species (Kindler et al. 2016; Petzold

et al. 2014) can serve here for comparison. Also, for

these species a wide range has been reported (pairwise

average divergences between species vary from 1.38%

to 16.54%), even though the low value of 1.38% between

the allopatric P. bechuanicus and P. upembae has been

suggested to indicate their conspecificity (Kindler et al.

2016). When only divergences of sympatric species are

considered, the values range between 2.49% and 15.35%

(Table 3). Yet, the lowest value refers to P. carinatus

and populations of P. rhodesianus that could actually

be conspecific with P. carinatus (Kindler et al. 2016).

August 2019 | Volume 13 | Number 2 | e184

Vamberger et al.

If this value is disregarded, the lowest value between

unambiguous sympatric species amounts to 5.43% (P.

subniger vs. P. upembae), and this value is much higher

than the divergence between clades A and B of P. sinuatus

(2.80%) found here.

In view of this relatively low value, we suggest

subspecies status for the Serrated Hinged Terrapins

from the southernmost and more northerly parts of the

distribution range. The name Sternotherus sinuatus

Smith, 1838 is clearly referable to the southern

subspecies, while the oldest name for the northern

subspecies is Sternothaerus bottegi Boulenger, 1895 (Fig.

1). Accordingly, the smaller-sized southern populations

represent the nominotypical subspecies Pelusios

sinuatus sinuatus (Smith, 1838), and the large-sized

northern subspecies is to be named Pelusios sinuatus

bottegi (Boulenger, 1895) nov. comb. Another name,

Pelusios sinuatus zuluensis Hewitt, 1927 (type locality:

Mzinene River, KwaZulu-Natal, South Africa) clearly

is a Junior synonym of the nominotypical subspecies.

A fourth name, Pelusios sinuatus leptus Hewitt, 1933

(type locality: Isoka, Zambia) can be identified with the

northern subspecies, and is thus a junior synonym of P.

Ss. bottegi.

This assessment is in line with the recent proposal to use

the subspecies category for naming lineages that qualify

for the genetic criteria of Evolutionarily Significant Units

(ESUs; Moritz 1994). Accordingly, subspecies should

correspond to distinct mtDNA lineages (except for cases

of mitochondrial capture), and they should be diagnosable

by nuclear genomic evidence. However, in contrast to

species, subspecies are genetically less divergent and

capable of large-scale gene flow with other subspecies.

Applying subspecies names for such lineages facilitates

communication within and beyond science, particularly

in legislation and conservation (Kindler and Fritz 2018).

In this vein, the recognition of two subspecies of P.

sinuatus not only reflects their genetic divergence but also

will contribute in the medium term to their conservation.

Currently, P. sinuatus is not considered to be imperiled

(IUCN category “Least Concern,” Rhodin et al. 2018).

However, in many African countries freshwater habitats

are increasingly threatened by progressing land use and,

consequently, the numbers of Serrated Hinged Terrapins

are dwindling. Furthermore, we propose to treat the

subclades Al—A3 within P. s. bottegi and subclades B1

and B2 within P. s. sinuatus as distinct Management

Units in the sense of Moritz (1994), 1.e., as populations

with significant mitochondrial divergence.

Conclusions

Serrated Hinged Terrapins (Pelusios sinuatus) show

concordant variation in mitochondrial and nuclear

marker genes, corresponding to two distinct genealogical

lineages in the southernmost and more _ northerly

parts of the distribution range. Each lineage displays

phylogeographic structuring. Terrapins representing the

two lineages differ also in body size, with individuals

from the northern and central parts of the distribution

reaching larger sizes than terrapins from the southern

parts. Considering the degree of genetic differentiation

Amphib. Reptile Conserv.

compared to other Pe/usios species, we conclude that the

two lineages should be regarded as distinct subspecies.

The nominotypical subspecies Pe/usios sinuatus sinuatus

(Smith, 1838) corresponds to populations in the south

(South Africa and parts of Botswana), and the resurrected

taxon Pelusios sinuatus bottegi (Boulenger, 1895) nov.

comb. to populations from the northern and central

distribution range. A contact zone of the two subspecies is

identified in the border region of Botswana, South Africa,

and Zimbabwe. The genetically differentiated population

clusters within each subspecies should be treated as

distinct Management Units. Further research is needed

to find out whether additional diagnostic morphological

characters for the two subspecies exist. In addition, denser

sampling would allow a fine-scale phylogeography for the

Species including an assessment of gene flow between the

two subspecies and the Management Units within each

subspecies. Such research could contribute significantly

to the development of long-term management plans for

this species. However, the current legislative situation

makes progress unlikely because multiple countries are

involved and obtaining sampling permits for biodiversity

research often becomes a major, if not insurmountable,

administrative obstacle.

Acknowledgements.—We dedicate this study to the

late Bill Branch (1946-2018), who donated many of

the samples that made this investigation possible. Other

material was collected during fieldwork in South Africa.

Fieldwork and sampling in South Africa were permitted

by the Limpopo Provincial Government (permit ZA/

Lp/80202) and Ezemvelo KwaZulu-Natal Wildlife

(permits OP 5139/2012, OP 526/2014, OP 839/2014,

OP 4374/2015, OP 4092/2016, OP 139/2017, and OP

4085/2017). Terrapins sampled in the field were released at

the collection site after taking blood samples 1n accordance

with methods approved by the Ethics Committee of

the University of the Western Cape (ethical clearance

number ScRiRC2008/39) and the Animal Care, Health,

and Safety in Research Ethics Committee (AnimCare)

of the North-West University (ethical clearance number

NWU-00372-16-A5). Additional material was donated

by Werner Conradie (Port Elizabeth Museum), Peter

Praschag (Turtle Island Graz), Louis du Preez (North-

West University), Pavel Siroky (University of Veterinary

and Pharmaceutical Sciences, Brno), and Krystal Tolley

(SANBI). Angela Gaylard, Samantha Mabuza, Zelna

Silcock, and Rheinhard Scholtz (SANParks) supported

us in our South African research project applications.

We thank the Ezemvelo KwaZulu-Natal Wildlife

Permits Office and the Limpopo Provincial Government

for permits to collect biological material, and private

landowners or property managers (Annemieke and

Hermann Miller of Lapalala Wilderness Area; Kwa

Nyamazane Conservancy and Pongola River Company)

for allowing sampling on their properties. Thanks for

access to museum specimens go to Christian Klug,

Torsten Scheyer (both PIUMZ), Mark-Oliver Rodel,

Frank Tillack (both ZMB), and José Rosado (MCZ).

This paper forms part of a VLIR-UOS TEAM project

(ZEIN21013PR396), co-funded by the Water Research

Commission of South Africa (Project K5-2185, Nico J.

August 2019 | Volume 13 | Number 2 | e184

Phylogeography of Pelusios sinuatus in East Africa

Smit). Edward C. Netherlands benefitted further from

financial assistance of the National Research Foundation

(NRF), a DAAD-NRF doctoral scholarship (Grant

108803), and the VLIR-UOS university scholarship (ID

0620854/Contract 000000076310). Opinions expressed

and conclusions arrived at are those of the authors

and are not necessarily to be attributed to the funding

bodies. Genetic investigations were conducted in the

Senckenberg Dresden Molecular Laboratory (SGN-

SNSD-Mol-Lab). Thanks for laboratory work go to Anja

Rauh and Anke Miller.

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Appendix 1. Studied material of Pe/usios sinuatus and outgroups and European Nucleotide Archive (ENA) accession numbers.

Accession numbers starting with LR correspond to sequences produced for the present study.

12S cyt b ND4 Cmos R35 Rag2 Provenance Latitude Longitude As

subclade

14050 | LR594053 | LR594111 | LR594157 | LRS94218 | LR594272 | LR594325 | Botswana: Goo- -22.58402 | 27.43988 Bl

Moremi Gorge

14051 | LR594054 | LR594112 | LR594158 | LR594219 | LR594273 | LR594326 | Botswana: Goo- -22.58402 | 27.43988 Bl

Moremi Gorge

14052 | LRs94055 | LR594113 | LR594159 | LR594220 | LR594274 | LR594327 | Botswana: Goo- -22.58402 | 27.43988 Bl

Moremi Gorge

14053 | LRs94056 | LR594114 | LR594160 | LR594221 | LR594275 | LR594328 | Botswana: Goo- -22.58402 | 27.43988 Bl

Moremi Gorge

14054 | LRs94057 | LR594115 | LR594161 | LR594222 | LR594276 | LR594329 | Botswana: Goo- -22.58402 | 27.43988 Bl

Moremi Gorge

14055 | LRs94058 | LR594116 | LR594162 | LR594223 | LR594277 | LR594330 | Botswana: Goo- -22.58402 | 27.43988

Moremi Gorge

14056 | LRs94059 | LR594117 | LR594163 | LR594224 | LR594278 | LR594331 | Botswana: Goo- -22.58402 | 27.43988

Moremi Gorge

5564 | FR716875 alae FR716984 | FR717028 | FR717076 | FR717121 | Botswana: Mashatu -22.212308 | 29.136038 Pa

Game Reserve

5565 | LR594060 LR594164 | LR594225 | LR594279 | LR594332 | Botswana: Mashatu -22.212308 | 29.136038

Game Reserve

Mozambique: Cabo

7044 | LR594061 n/a LR594165 | LR594226 | LR594280 | LR594333 | Delgado: wetland at -13,089139 | 40544583 A2

southern end of Pemba

drainage (Pemba)

Mozambique: Cabo

7045 | LRS94062 n/a LR594166 | LR594227 n/a Delgado: wetland at -13.089139 | 40.544583

southern end of Pemba

drainage (Pemba)

Mozambique: Cabo

7046 | LR594063 n/a LR594167 | LR594228 LR594334 | Delgado: wetland at -13.089139 | 40.544583

southern end of Pemba

drainage (Pemba)

Mozambique: Cabo

7047. | LRS94064 n/a LR594168 | LR594229 Delgado: wetland at -13.089139 | 40.544583

southern end of Pemba

drainage (Pemba)

Mozambique: Cabo

7049 | LRS94066 n/a LR594170 | LR594230 LR594335 | Delgado: wetland at -13,089139 | 40.544583

southern end of Pemba

drainage (Pemba)

17104 | LR594067 | LR594118 | LR594171 | LR594231 | LR594281 | LR594336 | Mozambique: Cabo -11.884827 } 40.460208

Delgado: Rio Diquide

LRS94282 Mozambique: Sofala

9891 LR594068 n/a LR594172 | LR594232 LR594337 | province, along road -20.923567 34.6662

LR594283 OR

August 2019 | Volume 13 | Number 2 | e184

Bales

Mozambique: Cabo

7048 | LRS594065 n/a LR594169 n/a n/a Delgado: wetland at -13.089139 | 40.544583 A2

southern end of Pemba

drainage (Pemba)

Amphib. Reptile Conserv. 52

Vamberger et al.

Appendix 1 (continued). Studied material of Pe/usios sinuatus and outgroups and European Nucleotide Archive (ENA) accession

numbers. Accession numbers starting with LR correspond to sequences produced for the present study.

12S cyt b ND4 Cmos R35 Rag2 Provenance Latitude Longitude a

subclade

Mozambique: Sofala

11100 LR594069 n/a LR594173 | LR594233 | LR594284 | LR594338 | province, along road -21.007317 | 34.539433 Al

428

LRS94285 Mozambique: Sofala:

6956 LR594070 | LR594119 | LR594174 | LR594234 LR594286 LR594339 | NE of Rio Save Game -20.933333 | 34.316667 Al

Reserve

Mozambique: Sofala:

6959 | LR594071 | LR594120 | LR594175 | LR594235 | LR594287 | LR594340 | NE of Rio Save Game -20.7425 34.586567 Al

Reserve

5215 | LRS594072 n/ LR594176 | LR594236 | LRS594288 | LR594341 Mozambique: -17.059513 | 38.699233 A2

Zambesia: Moebase

South A frica:

17003 | LR594073 | LR594121 | LR594177 | LR594237 | LR594289 | LR594342 | KwaZulu-Natal: -28.05752 | 32.29332 B2

Bonamanzi Game

Reserve

South A frica:

17004 | LR594074 | LR594122 | LR594178 | LR594238 | LR594290 | LR594343 | KwaZulu-Natal: -28.05752 | 32.29332 B2

Bonamanzi Game

Reserve

South A frica:

17005 | LR594075 | LR594123 | LR594179 | LR594239 | LR594201 | LR59434q | KwaZulu-Natal: -28.05752 | 32.29332 B2

Bonamanzi Game

Reserve

South A frica:

17006 | LR594076 } LR594124 | LR594180 | LR594240 | LR594292 | LRso434s | KwaZulu-Natal: -28.05752 | 32.29332 B2

Bonamanzi Game

Reserve

South Africa:

17010 | LR594077 | LR594125 | LR594181 n/a n/a nae) _ | eee al -28.145899 | 31.591881 B2

Bonamanzi Game

Reserve

South A frica:

9143. | LR594078 | LR594126 | LR594182 | LR594241 | LR594293 | LRs9o4346 | KwaZulu-Natal: -28.05809 | 32.29412 B2

Bonamanzi Game

Reserve, Waterlily Dam

South A frica:

14040 | LR594079 | LR594127 | LR594183 | LR594242 | LR594204 | LR594347 | KwaZulu-Natal: Jozini, | 57 591656 | 3.139094 B2

Kwa Nyamazane

Conservancy

South Africa:

KwaZulu-Natal:

16199 LR594080 | LR594128 | LR594184 | LR594243 | LR594295 LR594348 | Manyiseni region in -26.876389 | 32.011389 B2

Lebombo Mountains,

near Mabona

South Africa:

KwaZulu-Natal:

16200 LR594081 | LR594129 | LR594185 | LR594244 | LR594296 | LR594349 | Manyiseni region in -26.876389 | 32.011389 B2

Lebombo Mountains,

near Mabona

South Africa:

KwaZulu-Natal:

16202 LR594082 | LR594130 | LR594186 | LR594245 | LR594297 | LR594350 | Manyiseni region in -26.876389 | 32.011389 B2

Lebombo Mountains,

near Mabona

South A frica:

KwaZulu-Natal:

16203 LR594083 | LR594131 | LR594187 | LR594246 | LR594298 | LR594351 | Manyiseni region in -26.876389 | 32.011389 B2

Lebombo Mountains,

near Mabona

South Africa:

10614 LR594084 n/a LR594188 | LR594247 | LR594299 | LR594352 | KwaZulu-Natal: -26.885692 | 32.223672 B2

Ndumo Game Reserve

South Africa:

10615 LR594085 | LR594132 | LR594189 | LR594248 | LR594300 | LR594353 | KwaZulu-Natal: -26.874944 | 32.231997 B2

Ndumo Game Reserve

South Africa:

14041 LR594086 | LR594133 | LR594190 | LR594249 | LR594301 LR594354 | KwaZulu-Natal: -26.891275 32.299 B2

Ndumo Game Reserve

Amphib. Reptile Conserv. 53 August 2019 | Volume 13 | Number 2 | e184

Phylogeography of Pelusios sinuatus in East Africa

Appenidx 1 (continued). Studied material of Pe/usios sinuatus and outgroups and European Nucleotide Archive (ENA) accession

numbers. Accession numbers starting with LR correspond to sequences produced for the present study.

12S cyt b ND4 Cmos R35 Rag2 Provenance Latitude Longitude ae

subclade

South Africa:

13590 LR594087 | LR594134 | LR594191 | LR594250 | LR594302 | LR594355 KwaZulu-Natal: ; -26.865118 | 32.240964

Ndumo Game Reserve:

Mabayani

South Africa:

FR716876 FR716937 | FR716985 | FR717029 FR717077 FR717122 | KwaZulu-Natal: Phinda | -27.843744

Game Reserve

South Africa:

FR716877 | FR716938 | FR716986 | FR717030 | FR717078 FR717123 | KwaZulu-Natal: Phinda | -27.843744

Game Reserve

South Africa:

LR594088 | LR594135 | LR594192 | LR594251 | LR594303 | LR594356 | KwaZulu-Natal: St. -28.357158

Lucia: Crocodile Centre

South Africa:

LR594089 | LR594136 | LR594193 | LR594252 | LR594304 | LR594357 | KwaZulu-Natal: St. -28.357158

Lucia: Crocodile Centre

South Africa:

LR594090 | LR594137 | LR594194 | LR594253 | LR594305 LR594358 | KwaZulu-Natal: St. -28.357158

Lucia: Crocodile Centre

South Africa: Limpopo:

LR594091 | LR594138 | LR594195 | LR594254 | LR594306 | LR594359 | Hoedspruit: Bush -24.35039

Pub Inn

South Africa: Limpopo:

LR594092 | LR594139 | LR594196 | LR594255 | LR594307 | LR594360 | Hoedspruit: Bush -24.35039

Pub Inn

South Africa: Limpopo:

LR594093 | LR594140 | LR594197 | LR594256 | LR594308 | LR594361 | Hoedspruit: Bush -24.35039

Pub Inn

South Africa: Limpopo:

LR594094 | LR594141 | LR594198 | LR594257 | LR594309 | LR594362 | Hoedspruit: Bush -24.35039

Pub Inn

South Africa: Limpopo:

LR594095 | LR594142 | LR594199 | LR594258 | LR594310 | LR594363 | Hoedspruit: Bush -24.35039

Pub Inn

South Africa: Limpopo:

LR594096 | LR594143 | LR594200 | LR594259 | LR594311 LR594364 | Hoedspruit: Bush -24.35039

Pub Inn

South Africa: Limpopo:

LR594097_ | LR594144 | LR594201 | LR594260 | LR594312 | LR594365 | Hoedspruit: Bush -24.35039

Pub Inn

South Africa: Limpopo:

LR594098 | LR594145 | LR594202 | LR594261 | LR594313 LR594366 | Hoedspruit: Bush -24.35039

Pub Inn

South Africa: Limpopo:

LR594099 | LR594146 | LRS594203 | LR594262 | LRS594314 | LR594367 | Hoedspruit: Bush -24 35039

Pub Inn

LR594100 | LR594147 | LR594204 | LR594263 | LR594315 | LR594368 ea oe Limpopo: | _53 9392

LR594101 | LR594148 | LR594205 | LR594264 | LR594316 | LR594369 eae Limpopo: | _53 39392

South Africa: Limpopo:

LR594102 | LR594149 | LR594206 | LR594265 | LR594317 | LR594370 | Palabora Mining -24.018889

Company, near Loolo

Dam, S of Phalaborwa

South Africa: Limpopo:

LR594103 | LR594150 | LR594207 | LR594266 | LR594318 | LR594371 | Palabora Mining

Company, SE of

Phalaborwa

South Africa: Limpopo:

LR594104 | LR594151 | LR594208 n/a LR594319 | LR594372

Palabora Mining

LR594105 | LR594152 | LR594209

5216 32.335521

32.335521

17014 32.419512

17015 32.419512

17016 32.419512

17028 31.152019

17029 31.152019

17030 31.152019

17031 31.152019

17038

17039 31.152019

17040 31.152019

17041 31.152019

17042 31.152019

17061 28.29516

17068 28.29516

31.140833

Ne)

31.210556

Company: Cleveland

Nature Reserve, in

Olifants River near

picnic site

LR594267 | LR594320 | LR594373 | South Africa: Limpopo: | _53 98716

Vaalwater

-24.03056 31.19306

17073

Amphib. Reptile Conserv. 54 August 2019 | Volume 13 | Number 2 | e184

Vamberger et al.

Appenidx 1 (continued). Studied material of Pelusios sinuatus and outgroups and European Nucleotide Archive (ENA) accession

numbers. Accession numbers starting with LR feel to sequences produced for the present study.

s , mtDNA

LR594210

South Africa:

Mpumalanga: Kruger

13585 LR594106 | LR594153 LR594268 | LR594321 | LR594374 | National Park: -23.107365 | 31.439066 Bl

Shingwedzi River near

Shingwedzi Camp

South Africa:

Mpumalanga: Kruger

13586 LR594107 n/a LR594211 National Park: -23.107365 | 31.439066 Bl

Shingwedzi River near

Shingwedzi Camp

16214 | LR594108 | LR594154 | LR594212 | LR594269 | LR594322 | LR594375 Bae -3.34261 | 37319831

Tanzania: Manyara

16269 LR594109 | LR594155 | LR594213 | LR594270 | LR594323 | LR594376 | Region: Kikuletwa -3.443532 37.193393 A3

Hotsprings

Tanzania: Manyara

16270 LR594110 | LR594156 | LR594214 | LR594271 | LR594324 | LR594377 | Region: Kikuletwa -3.443532 37.193393 A3

Hotsprings

|_| Outgroups

Pelusios marani

Pelomedusa variabilis

Amphib. Reptile Conserv. 55 August 2019 | Volume 13 | Number 2 | e184

Amphib. Reptile Conserv.

Phylogeography of Pelusios sinuatus in East Africa

Melita Vamberger is a Slovenian herpetologist and evolutionary biologist working at the Senckenberg

Natural History Collections, Dresden, Germany. Melita studied Biology at the University of Ljubljana,

Slovenia, focusing on the natural history of the European Pond Turtle (Emys orbicularis). After

her diploma, Melita moved to Germany for her Ph.D. at the University of Leipzig, studying the

phylogeography and hybridization of two closely related freshwater turtle species (Mauremys caspica

and M. rivulata). Melita’s main interests are speciation, gene flow, adaptation, and evolution of different

turtle taxa using an integrative approach that combines genetic and ecological methods, especially in

the Western Palearctic and sub-Saharan Africa.

Margaretha D. Hofmeyr is Professor Emeritus at the Biodiversity and Conservation Biology

Department, University of the Western Cape, South Africa. Margaretha is an ecophysiologist by

training, and first studied large ungulates before switching to chelonians. Her ecophysiological

studies revealed that South African tortoises have many unique characteristics, which stimulated

further interest in their genetic diversity and systematics. Margaretha has published extensively on

the ecology and phylogeography of sub-Saharan tortoises and turtles, and she is closely involved in

conservation projects for threatened tortoises. This work resulted in her being awarded the 2015 Sabin

Turtle Conservation Prize. Margaretha is a member and Regional Vice-Chair for Africa of the IUCN/

SSC TFTSG and coordinated the 2014 and 2018 Red List Assessments for South African tortoises and

freshwater turtles.

Courtney A. Cook is a Senior Lecturer in the Water Research Group, Unit for Environmental Sciences

and Management, North-West University, South Africa. Courtney is a parasitologist focusing on

the biodiversity, taxonomy, and phylogeny of protozoan blood parasites of ectothermic vertebrates

(amphibians, reptiles, and fish), with several authored and co-authored scientific articles in this area. Her

M.Sc. and Ph.D. both focused exclusively on these parasites infecting tortoises of South Africa, which

inspired a keen interest in these animals, particularly with respect to their taxonomy and phylogeny,

both important aspects in understanding the associated host-parasite relationships. Courtney was

recently awarded a Y-rating by the South African National Research Foundation, identifying her as a

promising young researcher in her field from a global perspective.

Edward C. Netherlands is a dual Ph.D. candidate between the North-West University, South Africa,

and Katholieke Universiteit Leuven, Belgium. His Ph.D. forms part of the VLIR-UOS program for

the development of tools for the sustainable utilization and management of aquatic resources in

South Africa. Edward’s research interests focus on the molecular biology, ecology, and taxonomy

of herpetofauna and their associated parasites. He is also passionate about teaching the importance

of conservation to young minds and non-scientists. Ed has authored or co-authored several scientific

articles and a bilingual frog field guide (in English and Zulu). Edward also received the Research

Excellence for Next Generation Researchers Award as a final year Ph.D. Candidate from the National

Research Foundation in South Africa.

Uwe Fritz is the head of the Museum of Zoology, Senckenberg Natural History Collections in Dresden,

Germany, and Extraordinary Professor for zoology at the University of Leipzig, Germany. Uwe has

worked for many years on the taxonomy, systematics, and phylogeography of turtles and tortoises,

and has also studied snakes and lizards to a lesser extent. He is particularly interested in hybridization

patterns and gene flow in the contact zones of distinct taxa. Uwe has authored or co-authored numerous

scientific articles, mainly in herpetology, and has edited proceedings and books, among them the two

turtle volumes of the Handbook of Amphibians and Reptiles of Europe.

56 August 2019 | Volume 13 | Number 2 | e184

Official journal website:

amphibian-reptile-conservation.org

Amphibian & Reptile Conservation

13(2) [Special Section]: 57-60 (e185).

Mind the gap—ls the distribution range of Pelomedusa

galeata really disjunct in western South Africa?

‘Melita Vamberger, ‘Paula Ribeiro Anunciagao, *Margaretha D. Hofmeyr, and ‘Uwe Fritz

'Museum of Zoology, Senckenberg Natural History Collections Dresden, A.B. Meyer Building, 01109 Dresden, GERMANY ?Chelonian Biodiversity

and Conservation, Department of Biodiversity and Conservation Biology, University of the Western Cape, Bellville 7535, SOUTH AFRICA

Abstract.—Records from the putative gap in the distribution range of Pelomedusa galeata in western South

Africa provide evidence for the occurrence of helmeted terrapins in those areas. Further research is needed to

reveal the genetic and taxonomic identity of these populations.

Keywords. helmeted terrapin, Northern Cape Province, Pelomedusidae, Testudines, turtle

Citation: Vamberger M, Anunciagao PR, Hofmeyr MD, Fritz U. 2019. Mind the gap—ts the distribution range of Pelomedusa galeata really disjunct in

western South Africa? Amphibian & Reptile Conservation 13(2) [Special Section]: 57-60 (e185).

tion 4.0 International (CC BY 4.0): https://creativecommons.org/licenses/by/4.0/], which permits unrestricted use, distribution, and reproduction in any

medium, provided the original author and source are credited. The official and authorized publication credit sources, which will be duly enforced, are

as follows: official journal title Amphibian & Reptile Conservation; official journal website: amphibian-reptile-conservation.org.

Received: 28 February 2019; Accepted: 22 March 2019; Published: 20 August 2019

Helmeted terrapins have a wide distribution across sub-

Saharan Africa and the southwestern Arabian Peninsula.

Together with their sister taxon, the African hinged

terrapins (Pelusios), helmeted terrapins constitute the

side-necked turtle family Pelomedusidae (TTWG 2017).

While it was assumed for decades that all helmeted

terrapins are conspecific (Boycott and Bourquin 2008;

Branch 2008; Ernst and Barbour 1989; Wermuth and

Mertens 1961, 1977), several investigations revealed

deep genetic divergences between geographically

coherent populations, which resemble or exceed the

divergences of distinct Pe/usios species (Fritz et al. 2011,

2014; Petzold et al. 2014; Vamberger et al. 2018; Vargas-

Ramirez et al. 2010; Wong et al. 2010). This resulted in

the formal recognition of no less than 10 distinct species

(Petzold et al. 2014; TTWG 2017; Vamberger et al. 2018).

In addition to these, a minimum of five unnamed species

are thought to exist, which are characterized by similar

genetic divergences but otherwise only insufficiently

known (Fritz et al. 2015; Nagy et al. 2015; Petzold et

al. 2014; Vamberger et al. 2018; Vargas-Ramirez et al.

2016).

In the Southern African region south of the Cunene

and Zambezi Rivers, two species are known to occur,

the South African Helmeted Terrapin Pelomedusa

galeata (Schoepff, 1792) and the Common Helmeted

Terrapin Pelomedusa subrufa (Bonnaterre, 1789) (Fritz

et al. 2015; Petzold et al. 2014; Vamberger et al. 2018).

Pelomedusa galeata is distributed in most of South Africa

and replaced in the countries north of South Africa by

P. subrufa, which enters also northeastern South Africa

(provinces of Limpopo and Mpumalanga). In these

Correspondence. * wve.fritz@senckenberg.de

Amphib. Reptile Conserv.

provinces, the distribution ranges of the two species

abut and the closest records of P. galeata and P. subrufa

are separated only by 80 km, so that overlapping ranges

seem possible (Vamberger et al. 2018). Pelomedusa

galeata shows pronounced phylogeographic structuring,

with two genetically deeply divergent groups in the east

and west of South Africa that most likely represent two

distinct species. The eastern group is phylogeographically

structured, with three parapatric subgroups (Fritz et al.

2015; Petzold et al. 2014; Vamberger et al. 2018).

Detailed distribution maps for helmeted terrapins show

a patchy range for Southern Africa, with large putative

gaps in western South Africa, southern Mozambique,

southern Namibia, and most of Botswana (Boycott 2014;

Boycott and Bourquin 2008; TTWG 2017). Except

for southern Mozambique, these regions are very arid,

suggestive of pessimal conditions for a freshwater turtle

like the helmeted terrapin. However, P. subrufa is known

to cope with year-long drought. In Namibia (Omaheke),

terrapins of this species may evidently survive up to six

years burrowed in the soil (Petzold et al. 2014) and surface

only after the rare rainfalls. Boycott and Bourquin (2008)

suggested that helmeted terrapins take advantage of man-

made farm dams and expanded their range into otherwise

unsuitable regions, including semi-desert. However,

records for P subrufa from the mouths of temporary

streams in the Namib Desert may well represent natural

occurrences of terrapins washed downstream during the

rare floods there (A. Schleicher, pers. comm. ).

One large distribution gap is located in western

South Africa and concerns P. galeata. It more or less

separates the two genetically deeply divergent groups of

this species (Vamberger et al. 2018). During fieldwork

in October and November 2018, the first two authors

August 2019 | Volume 13 | Number 2 | e185

Distribution of Pelomedusa galeata in South Africa

ere Se A . ae

Fig. 1. Pelomedusa galeata Rott if Ratelfontein farm near Calvinia observed directly after eal 16 February 2019. For the

location of the farm, see Fig. 2 (locality 1). Photos: C.A. van Niekerk.

Northern Cape

Western Cape

North West

Free State

Fig. 2. Distribution range of Helmeted Terrapins (shaded in grey), with our records of Pelomedusa galeata in South Africa (white

circles). New records of P. galeata in or close to the putative distribution gap: 1 — Nineteen turtles at Ratelfontein farm, near

Calvinia (16 February 2019), 2 — Observations of locals at Williston, 3 — Near Carnarvon (shell, collected 25 October 2018), 4— One

terrapin near Beaufort West (4 March 2017), 5 — Two terrapins near Griekwastad (28 October 2018). Inset: Pelomedusa galeata

from the Ratelfontein farm. Photo: C.A. van Niekerk.

had the opportunity to examine parts of this putative

distribution gap for the presence of helmeted terrapins. In

addition to direct observations, interviews with farmers

and locals contributed further information. It is common

knowledge there that helmeted terrapins are present but

very scarce. They are seen only after the rare rainfall

events, when the terrapins are walking to waterholes (Fig.

1). Together with our records substantiated by specimens,

Amphib. Reptile Conserv.

this provides the first evidence for the occurrence of P.

galeata in the central part of the Northern Cape Province

of South Africa (Fig. 2). We assume that the helmeted

terrapin has an even wider distribution and also occurs

northwards to Namibia, and that a distribution gap does

not exist at all. Pelomedusa are very elusive animals

in arid regions, and we presume that the putative gap

reflects not a real absence of helmeted terrapins but

August 2019 | Volume 13 | Number 2 | e185

Vamberger et al.

a lack of records. Further research is needed to reveal

which genetic lineage of P galeata occurs in this area

and whether there is a contact zone of the two lineages

currently identified with P. galeata.

Acknowledgements.-We would like to thank C.A. and

Berno van Niekerk from Ratelfontein farm, Adam van

Greunen, Isak Dreyer, Pieter and Elmarie Naude from

Die Ark, Rodney Bartholomew, Haas van Niekerk,

Bertus and Roux Steenkamp, Nadia and Abri van Zyl,

Adriaan Jordaan, Nick Telford, and Krystal Tolley for

all the valuable information, pictures, and help during

fieldwork. Fieldwork of M.V. and P.R.A. was supported

by the DGHT (Deutsche Gesellschaft fir Herpetologie

und Terrarienkunde) and the BCG (British Chelonian

Group). M.D.H. was supported by the Mapula Trust.

Fieldwork was permitted by the North West Province

(NW 6124/10/2018) and Northern Cape Province

(0729/2018 and 0730/2018).

Literature Cited

Boycott RC. 2014. Pelomedusa subrufa (Bonnaterre,

1789). Pp. 54 In: Atlas and Red List of the Reptiles

of South Africa, Lesotho and Swaziland. Suricata 1.

Editors, Bates MF, Branch WR, Bauer AM, Burger M,

Marais J, Alexander GJ, de Villiers MS. South African

National Biodiversity Institute (SANBI) Publishing,

Pretoria, South Africa. xviii, 486 p.

Boycott RC, Bourquin O. 2008. Pelomedusa subrufa

(Lacépede 1788) helmeted turtle, helmeted

terrapin. Pp. 007.1-007.6 In: Conservation Biology

of Freshwater Turtles and Tortoises: a Compilation

Project of the IUCN/SSC Tortoise and Freshwater

Turtle Specialist Group. Chelonian Research

Monographs 5. Editors, Rhodin AGJ, Pritchard PCH,

van Dijk PP, Saumure RA, Buhlmann KA, Iverson

JB. Chelonian Research Foundation, Lunenburg,

Massachusetts, USA. Loose-leaf (pagination

variable).

Branch WR. 2008. Tortoises, Terrapins & Turtles of

Africa. Struik, Cape Town, South Africa. 128 p.

Ernst CH, Barbour RW. 1989. Turtles of the World.

Smithsonian Institution Press, Washington, D.C.,

USA, and London, United Kingdom. xu, 313 p.

Fritz U, Branch WR, Hofmeyr MD, Maran J, Prokop H,

Schleicher A, Siroky P, Stuckas H, Vargas-Ramirez

M, Vences M, et al. 2011. Molecular phylogeny of

African hinged and helmeted terrapins (Testudines:

Pelomedusidae: Pelusios and Pelomedusa). Zoologica

Scripta 40: 115-125.

Amphib. Reptile Conserv.

Fritz U, Kehlmaier C, Mazuch T, Hofmeyr MD, du

Preez L, Vamberger M, Voros J. 2015. Important new

records of Pelomedusa species for South Africa and

Ethiopia. Vertebrate Zoology 65: 383-389.

Fritz U, Petzold A, Kehlmaier C, Kindler C, Campbell

P, Hofmeyr MD, Branch WR. 2014. Disentangling

the Pelomedusa complex using type specimens and

historical DNA (Testudines: Pelomedusidae). Zootaxa

3795: 501-522.

Nagy ZT, Kielgast J; Moosig M, Vamberger M, Fritz

U. 2015. Another candidate species of Pelomedusa

(Testudines: Pelomedusidae) from the Democratic

Republic of the Congo? Salamandra 51: 212-214.

Petzold A, Vargas-Ramirez M, Kehlmaier C, Vamberger

M, Branch WR, du Preez L, Hofmeyr MD, Meyer L,

Schleicher A, Siroky P, etal. 2014. Arevision of African

helmeted terrapins (Testudines: Pelomedusidae:

Pelomedusa), with descriptions of six new species.

Zootaxa 3795: 523-548.

TTWG (Turtle Taxonomy Working Group), Rhodin AGJ,

Iverson JB, Bour R, Fritz U, Georges A, Shaffer HB,

van Dik PP. 2017. Turtles of the World. Annotated

Checklist and Atlas of Taxonomy, Synonymy,

Distribution, and Conservation Status. 8th edition.

Chelonian Research Monographs 7. Chelonian

Research Foundation, Lunenburg, Massachusetts,

USA. 292 p.

Vamberger M, Hofmeyr MD, Ihlow F, Fritz U. 2018.

In quest of contact: phylogeography of helmeted

terrapins (Pelomedusa galeata, P. subrufa sensu

stricto). PeerJ 6: e4901.

Vargas-Ramirez M, Petzold A, Fritz U. 2016. Distribution

modelling and conservation assessment for helmeted

terrapins (Pelomedusa spp.). Salamandra 52: 306—

316.

Vargas-Ramirez M, Vences M, Branch WR, Daniels

SR, Glaw F, Hofmeyr MD, Kuchling G, Maran J,

Papenfuss T, Siroky P, et al. 2010. Deep genealogical

lineages in the widely distributed African Helmeted

Terrapin: evidence from mitochondrial and nuclear

DNA _ (Testudines: Pelomedusidae: Pelomedusa

subrufa). Molecular Phylogenetics and Evolution 56:

428-440.

Wermuth H, Mertens R. 1961. Schildkréten, Krokodile,

Briickenechsen. VEB Gustav Fischer Verlag, Jena,

Germany. XX VI, 422 p.

Wermuth H, Mertens R. 1977. Testudines, Crocodylia,

Rhynchocephalia. Das Tierreich 100: 1-174.

Wong RA, Fong JJ, Papenfuss TJ. 2010. Phylogeography

of the African Helmeted Terrapin, Pelomedusa

subrufa. genetic structure, dispersal, and human

introduction. Proceedings of the California Academy

of Sciences 61: 575-585.

August 2019 | Volume 13 | Number 2 | e185

Amphib. Reptile Conserv.

Distribution of Pelomedusa galeata in South Africa

Melita Vamberger is a Slovenian herpetologist and evolutionary biologist working at the Senckenberg

Natural History Collections, Dresden, Germany. Melita studied Biology at the University of Ljubljana,

Slovenia, focusing on the natural history of the European Pond Turtle (Emys orbicularis). After her

diploma, she moved to Germany for her Ph.D. at the University of Leipzig on the phylogeography

and hybridization of two closely related freshwater turtle species (VMauremys caspica and M. rivulata).

Melita’s main interests are speciation, gene flow, adaptation, and evolution of different turtle taxa using an

integrative approach that combines genetic and ecological methods, especially in the Western Palearctic

and sub-Saharan Africa.

Paula Ribeiro Anunciac4o is a Brazilian ecologist and herpetologist. She is mainly interested in the

consequences of human disturbance for tropical amphibian communities. Paula studied biology at the

Federal University of Alfenas, Minas Gerais, Brazil. There, she also earned her master’s degree in

Ecology and examined the influence of matrix types and habitat fragmentation on the amphibian diversity

of the Atlantic rainforest. Paula earned her Ph.D. in Applied Ecology at the Federal University of Lavras,

Minas Gerais, Brazil, in 2018. For her Ph.D., she studied the relationships of land use change, climate

change, and taxonomic and functional richness of amphibians, which included some months of work at

the Senckenberg Natural History Collections, Dresden, Germany.

Margaretha D. Hofmeyr is Professor Emeritus at the Biodiversity and Conservation Biology

Department, University of the Western Cape, South Africa. She is an ecophysiologist by training and first

studied large ungulates before switching to chelonians. Her ecophysiological studies revealed that South

, African tortoises have many unique characteristics, which stimulated her interest in their genetic diversity

and systematics. Margaretha has published extensively on the ecology and phylogeography of sub-

Saharan tortoises and turtles, and she is closely involved in conservation projects that focus on threatened

tortoises. This work resulted in her being awarded the 2015 Sabin Turtle Conservation Prize. Margaretha

is amember, and Regional Vice-Chair for Africa, of the IUCN/SSC TFTSG and she coordinated the 2014

and 2018 Red List Assessments for South African tortoises and freshwater turtles.

Uwe Fritz is the head of the Museum of Zoology, Senckenberg Natural History Collections at Dresden,

Germany, and Extraordinary Professor for Zoology at the University of Leipzig, Germany. He has worked

for many years on the taxonomy, systematics, and phylogeography of turtles and tortoises, and also

studied to a lesser extent snakes and lizards. Uwe is particularly interested in hybridization patterns and

gene flow in contact zones of distinct taxa. He has authored or co-authored numerous scientific articles,

mainly in herpetology, and has also edited proceedings and books, among them the two turtle volumes of

the Handbook of Amphibians and Reptiles of Europe.

60 August 2019 | Volume 13 | Number 2 | e185

Official journal website:

amphibian-reptile-conservation.org

Amphibian & Reptile Conservation

13(2) [Special Section]: 61-67 (e195).

Geographic range extension of Speke’s

Hinge-back Tortoise Kinixys spekii Gray, 1863

1*Flora lhlow, 2**°Harith M. Farooq, °Vaclav Gvozdik, ‘Margaretha D. Hofmeyr, ®°Werner Conradie,

‘Patrick D. Campbell, “James Harvey, ‘*Luke Verburgt, and ‘Uwe Fritz

'Museum of Zoology, Senckenberg Dresden, A. B. Meyer Building, 01109 Dresden, GERMANY *Faculty of Natural Sciences, Lurio University, Pemba

958, MOZAMBIQUE Gothenburg Global Biodiversity Centre, 40530 Gothenburg, SWEDEN *Department of Biology and CESAM, University of

Aveiro, 3810-193 Aveiro, PORTUGAL *Department of Biological and Environmental Sciences, University of Gothenburg, 40530 Gothenburg, SWEDEN

°Institute of Vertebrate Biology of the Czech Academy of Sciences, 60365 Brno, CZECH REPUBLIC ‘Chelonian Biodiversity and Conservation,

Department of Biodiversity and Conservation Biology, University of the Western Cape, Bellville 7535, SOUTH AFRICA *Port Elizabeth Museum

(Bayworld), P.O. Box 13147, Humewood 6013, SOUTH AFRICA °School of Natural Resource Management, George Campus, Nelson Mandela

University, George 6530, SOUTH AFRICA '°Department of Life Sciences, Natural History Museum, London, UNITED KINGDOM ''41 Devonshire

Avenue, Howick, 3290, SOUTH AFRICA '*Department of Zoology and Entomology, University of Pretoria, Pretoria, 0001, SOUTH AFRICA

Abstract.—Kinixys spekii has a wide distribution range across sub-Saharan Africa, having been reported from

Angola, Botswana, Burundi, the Democratic Republic of the Congo, eSwatini, Kenya, Malawi, Mozambique,

Namibia, South Africa, Tanzania, Zambia, and Zimbabwe. Kinixys spekii inhabits savannah and dry bushveld

habitats and was previously considered an inland species. However, recent records suggest a more extensive

geographical distribution. Here, we provide genetically verified records for Angola, South Africa, and

Mozambique, and discuss reliable sightings for Rwanda. These new records extend the range significantly to

the east and west, and provide evidence for the occurrence of this species along the coast of the Indian Ocean

in South Africa and Mozambique.

Keywords. Africa, Angola, chelonians, distribution, Mozambique, Reptilia, Rwanda, Testudinidae

Citation: lhlow F, Farooq HM, Gvozdik V, Hofmeyr MD, Conradie W, Campbell PD, Harvey J, Verburgt L, Fritz U. 2019. Geographic range extension

of Speke’s Hinge-back Tortoise Kinixys spekii Gray, 1863. Amphibian & Reptile Conservation 13(2) [Special Section]: 61-67 (e195).

4.0 International (CC BY 4.0): https://creativecommons.org/licenses/by/4.0/], which permits unrestricted use, distribution, and reproduction in any

medium, provided the original author and source are credited. The official and authorized publication credit sources, which will be duly enforced, are

as follows: official journal title Amphibian & Reptile Conservation; official journal website: amphibian-reptile-conservation.org.

Received: 17 July 2019; Accepted: 28 September 2019; Published: 6 November 2019

The genus Kinixys currently comprises eight species

(Kindler et al. 2012; TTWG 2017): K. belliana Gray,

1830; K. erosa (Schweigger, 1812); K. homeana

Bell, 1827; K. lobatsiana Power, 1927; K. natalensis

Hewitt, 1935; K. nogueyi (Lataste, 1886); K. spekii

Gray, 1863; and K. zombensis Hewitt, 1931. Two of

these species are confined to rainforest habitats (K.

erosa, K. homeana), while one is restricted to north-

western Africa (K. nogueyi), and the remaining five

occupy savannah and forest habitats in eastern and

south-eastern Africa.

Speke's Hinge-back Tortoise, Kinixys spekii, has an

extensive geographical distribution range, spanning

twelve countries, from southern Kenya southward to

eSwatini (formerly Swaziland), southern Mozambique,

and north-eastern South Africa, where it reaches its

southernmost limit (Boycott and Bourquin 2000; Branch

et al. 1995; Broadley 1989a; Spawls et al. 2004, 2018;

TTWG 2017). The species’ westward range extends

across Zimbabwe, Zambia, and northern Botswana

into the Zambezi (formerly Caprivi) region of Namibia

(Broadley 1989a, 1993; Jacobsen et al. 1986; Pienaar

et al. 1983; TTWG 2017). According to Broadley

(1989a,b, 1993), K. spekii is confined to the inland parts

of southern and central Africa, inhabiting the eastern

plateau slopes, while the range of K. zombensis extends

along the East African coastal plain from Kenya to the

KwaZulu-Natal Province of South Africa. However,

a few records from Kenya (Watamu, 3.34250°S,

40.02740°E; in the vicinity of Kilifi, voucher specimen

in the collection of the Yale Peabody Museum of

Natural History YPM HERR 014516) suggest that the

range of K. spekii reaches the northern coastal areas as

well. Since some photographs of tortoises from Watamu

shown in Spawls et al. (2004, 2018) morphologically

resemble K. zombensis rather than K. spekii, this record

requires verification.

In terms of habitat, K. spekii has been recorded from

savannah, tropical bushveld, tropical savannah, sour

bushveld, and the thornveld of the Lebombo Plateau

Correspondence. flora.ihlow@senckenberg.de (*F1), harithmorgadinho@gmail.com (HMB), vaclav.gvozdik@ivb.cz (VG),

mdhofmeyr@gmail.com (MDH), werner@bayworld.co.za (WC), p.campbell@nhm.ac.uk (PDC), info@harveyecological.co.za (JH),

luke@enviro-insight.co.za (LV), uwe.fritz@senckenberg.de (UF)

Amphib. Reptile Conserv.

November 2019 | Volume 13 | Number 2 | e195

Distribution of Kinixys spekii in Africa

(Boycott 2001; Boycott and Bourquin 2000; Branch

2008). According to Broadley (1989a), this species

prefers moist savannah woodlands, such as Miombo and

Mopane (woodlands dominated by Brachystegia and

Colophospermum species, respectively), but also occurs

in drier deciduous woodlands and thickets dominated

by Vachellia (until recently Acacia; Kyalangalilwa et al.

2013) and Commiphora in the north-eastern part of its

range (Broadley 1989a).

For the present contribution, records for K. spekii

were compiled from the scientific literature and

museum collections, and supplemented with a few

selected sightings from the online Virtual Museum

Database Naturalist (https://www.inaturalist.org) to

discuss the distribution range of K. spekii. Several new

and genetically verified records from Angola, South

Africa, and coastal Mozambique are also presented,

which extend the species’ known distribution range

considerably. Genetic verification relied on an mtDNA

sequence coding for the partial NADH dehydrogenase

subunit 4 (ND4) and adjacent tRNAs.

Unfortunately, online databases and data aggregators

are often compromised by incorrect identifications and

outdated taxonomy, and either provide no photographic

vouchers or ones that are unsuitable for facilitating

verification before using the data. Nevertheless, two

photographic vouchers deposited with Naturalist

(https://www. inaturalist.org/observations/18255494,

https://www. inaturalist.org/observations/1047117) could

clearly be assigned to K. spekii, based on characteristic

coloration patterns and shell shape. These records

provide evidence that the species occurs as far north-

west as Nyagatare (1.42321°S, 30.63027°E) and

Akagera (1.55162°S, 30.60760°E) in Rwanda, which

is in accordance with Spawls et al. (2004, 2018), who

reported isolated records for K. spekii from Akagera, the

Ruzizi Plain, and the southern Kerio Valley in eastern

Rwanda. To the east, two genetically verified K. spekii

(Museum of Zoology, Senckenberg Dresden, Tissue

Collection: MTD 17106, 17107; European Nucleotide

Archive accession numbers: LR723010, LR723011)

were sampled and released by Luke Verburgt in February

2014 on the Afungi Peninsula, Cabo Delgado Province,

coastal Mozambique (10.81939°S, 40.54842°E; but see

below). In addition, four genetically verified adult K.

spekii (MTD 20463, 20464; LR723016, LR723017) were

sampled and released by Harith M. Faroog within a 10 km

radius of the Lurio University in Pemba, Cabo Delgado

Province, Mozambique (12.97540°S, 40.57083°E) in

2014. An adult female (12.97615°S, 40.10205°E) and

an adult male tortoise (12.99333°S, 39.94861°E) were

sampled and released by William R. Branch in March

2017 in Ancuabe, Cabo Delgado Province, Mozambique,

and both were genetically verified to represent K. spekii

(MTD 20463-20464; LR723016-7). In the west, a genetic

sample (SANBI 2126; LR723018) collected by Thomas

Branch in October 2008 shows that K. spekii also occurs

Amphib. Reptile Conserv.

near Saurimo, Lunda Sul Province, Angola (9.39694°S,

20.43194°E).

With the exception of the tortoises from the Afungi

Peninsula, all specimens could unambiguously be

identified morphologically as K. spekii based on the

following characteristics: beak unicuspid, carapace with

well-developed hinge, carapace distinctly depressed,

and posterior marginal scutes not recurved or serrated.

Previous records for Mozambique were limited to the

southwest (Boycott and Bourquin 2000), the vicinity

of Ressano Garcia, southern Mozambique (Broadley

1989b), and the Maputo Elephant Reserve situated along

the coast of southern Mozambique (voucher specimen in

the collection of the Ditsong National Museum of Natural

History TM 41761; Broadley 1993; Fig. 1). To the south,

samples collected in the KwaZulu-Natal Province, South

Africa, were genetically verified as K. spekii (MTD

13594 from Mkhuze Game Reserve; LR723019; MTD

7457 from the vicinity of Mtubatuba; HE662316).

Previously, K. spekii was only known from the extreme

northern border of the province, adjacent to eSwatini and

Mozambique (Bourquin 2004; Boycott 2014).

The abovementioned records enlarge the known

geographical distribution range of K. spekii, but also

demonstrate that the species’ distribution range is still

incompletely known. Unfortunately, morphological

traits overlap between Kinixys species, making

species identification in potential zones of sympatry

extremely difficult. For instance, K. spekii co-occurs

with K. zombensis in the Maputo Elephant Reserve in

Mozambique, and the Ndumo Game Reserve in South

Africa. In the Waterberg area of South Africa K. spekii

is found close to K. lobatsiana populations, and it was

recorded together with K. natalensis in the vicinity of

Jameson’s Drift, in the Lebombo Mountains in KwaZulu-

Natal as well as in the area of Hoedspruit. In these areas,

hybridization is possible, which further complicates

identification. For example, the two tortoises collected

from the Afungi Peninsula in northern Mozambique

morphologically resemble K. zombensis (Fig 3;

domed carapace, radial coloration pattern), but their

mtDNA sequences match those of K. spekii, suggesting

hybridization. Additional genetic studies using nuclear

genes are required to verify their putative hybrid status.

However, if there is no evidence for a hybrid identity

in these tortoises, then the morphological characters

thought to be diagnostic for discerning K. spekii and K.

zombensis would be seriously challenged. Moreover,

the characteristic coloration patterns commonly used to

distinguish between Kinixys species tend to fade with

age, rendering older tortoises more or less uniformly

colored (Branch 2008; Broadley 1993; Fig. 2). Hence,

hinge-back tortoises are frequently misidentified. Genetic

verification of specimens which were morphologically

determined by renowned African herpetologists revealed

misidentification rates ranging from 2% for K. zombensis

to 66% for K. natalensis (F. Ihlow and U. Fritz unpub.

November 2019 | Volume 13 | Number 2 | e195

lhlow et al.

Legend

A\ Type specimens

e Sightings & records

@ Genetically verified

eet NN

4 7

a ZIMBABWE

‘Cale

al <a

500 km

Fig. 1. Known distribution of Kinixys spekii. Range according to TTWG (2017) is displayed as green shaded area. Open circles refer

to iNaturalist observations, while solid green circles represent literature records and specimens deposited in scientific collections.

Solid red circles correspond to genetically verified records, and triangles to name-bearing type specimens of Kinixys spekii and its

synonyms.

data). While older, uniformly colored K. /obatsiana are

mainly confused with K. spekii and vice versa, young

K. spekii are frequently confused with K. natalensis

(Fig. 2). The most reliable morphological trait allowing

distinction between K. natalensis and young K. spekii is

the tricuspid beak of K. natalensis, whereas K. spekii has

a unicuspid beak. Old uniformly colored K. /obatsiana

can be distinguished from K. spekii based on the posterior

Shell rim, which is serrated in K. /obatsiana and smooth

in K. spekii (Fig. 2).

The high misidentification rates show that the

established morphological characters for species

determination are insufficient and call into question the

reliability of published records, photographic vouchers,

Amphib. Reptile Conserv.

and even the identification of specimens (including

name-bearing type specimens) housed in scientific

collections. Given that the putative distribution ranges

(TTWG 2017), which represent an essential tool for

the conservation and management of these species, also

largely rely on collection databases and morphologically

identified individuals, these should be treated with

caution.

To ensure correct species identification of challenging

specimens, molecular genetic verification is strongly

recommended until more robust morphological

characters have been revealed. Photographic vouchers

should include dorsal, ventral, and lateral views to

facilitate accurate species identifications. For K.

November 2019 | Volume 13 | Number 2 | e195

Distribution of Kinixys spekii in Africa

eee

Vey PN Z,

Zee

Fig. 2. Top: Lateral views of adult Kinixys lobatsiana (left) and K.

th OE PME Ny oat Pek tsk all

spekii (right). Center: Ventral views of young (SCL 106 mm)

* Cipla “igh? wel

and adult (SCL 161 mm) K. /obatsiana (left) and young (SCL 131 mm) and adult (SCL 151 mm) K. spekii (right). Note the strongly

serrated posterior marginal scutes in K. /obatsiana compared to the smooth carapace rim in K. spekii. Bottom: Lateral views of adult

K. natalensis (left) and young K. spekii (right). Photos: James Harvey and Flora lhlow.

natalensis additional voucher photographs showing the

tricuspid beak should be taken.

Acknowledgements.—We are grateful to the

following collections, and their curators and managers,

for providing data and photographs of specimens or

access to their collections: Shiela Broadley (National

Museum Zimbabwe), Lauretta Mahlangu (Ditsong

National Museum of Natural History), Garin Cael (Royal

Museum for Central Africa), and José Rosado (Museum of

Comparative Zoology). We further thank Marius Burger

and Steve Spawls, who provided data, pictures, or genetic

samples, and the locals who kindly permitted sampling on

Amphib. Reptile Conserv.

their properties. We are grateful to Anders G.J. Rhodin for

sharing a dataset of occurrence records compiled for the

latest TFTSG checklist. In addition, FI thanks Anja Rauh

and Anke Muller (Senckenberg Dresden) for assistance

during laboratory work. All genetic analyses were done

at the molecular genetic laboratories of the Museum of

Zoology (SGN-SNSD-Mol-Lab), Senckenberg Dresden.

FI profited from a Margarethe Koenig scholarship from

the Zoological Research Museum Alexander Koenig.

In addition, research by FI is supported by the German

Science Foundation (DFG IH 133/1—1). Fieldwork was

partly supported through the Mapula Trust awarded to

MDH.

November 2019 | Volume 13 | Number 2 | e195

lhlow et al.

hitmen Oe

Fig. 3. Top: Lateral views of the putative hybrids from the Afungi Penins

if i P ~ ~ wes § ‘ ~~ =]

@ ~ tS ae

BED f : Ky ;

ula, Cabo Delgado Province, Mozambique, which have

mtDNA sequences of Kinixys spekii but morphologically resemble K. zombensis. Bottom: Lateral views of genetically verified K.

zombensis from KwaZulu-Natal Province, South Africa. Photos: Luke Verburgt and Flora Ihlow.

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tortoise. Pp. 49-52 In: The Conservation Biology

of Tortoises. Editors, Swingland IR, Klemens MW.

IUCN, Gland, Switzerland. 203 p.

Broadley DG. 1993. A review of the southern African

species of Kinixys Bell (Reptilia: Testudinidae).

Annals of the Transvaal Museum 36: 41-52.

Jacobsen NHG, Newbery RE, Petersen WL. 1986.

A Checklist of the Herpetofauna of the Transvaal

Provincial Nature Reserves. ‘Transvaal Nature

Conservation Division, Pretoria, South Africa. 38 p.

Kindler C, Branch WR, Hofmeyr MD, Maran J, Siroky

P, Vences M, Harvey J, Hauswaldt JS, Schleicher

A, Stuckas H et al. 2012. Molecular phylogeny of

African hinge-back tortoises (Kinixys): implications

for phylogeography and taxonomy (Testudines:

Testudinidae). Journal of Zoological Systematics and

Evolutionary Research 50: 192-201.

Kyalangalilwa B, Boatwright JS, Daru BH, Maurin

O, van der Bank M. 2013. Phylogenetic position

and revised classification of Acacia s.l. (Fabaceae:

November 2019 | Volume 13 | Number 2 | e195

Distribution of Kinixys spekii in Africa

Mimosoideae) in Africa, including new combinations Field Guide to East African Reptiles. Bloomsbury

in Vachellia and Senegalia. Botanical Journal of the Publishing PLC, London, United Kingdom. 624 p.

Linnean Society 172: 500-523. TTWG [Turtle Taxonomy Working Group: Rhodin AGJ,

Pienaar U de V, Haacke WO, Jacobsen NHG. 1983. The Iverson JB, Bour R, Fritz U, Georges A, Shaffer HB,

Reptiles of the Kruger National Park. National Parks van Dik PP]. 2017. Turtles of the World: Annotated

Board South Africa, Pretoria, South Africa. 236 p. Checklist and Atlas of Taxonomy, Synonymy,

Spawls S, Howell K, Drewes C, Ashe J. 2004. A Field Distribution, and Conservation Status. 8 Edition.

Guide to the Reptiles of East Africa. Gardners Books, Chelonian Research Monographs 7. Chelonian

Eastbourne, United Kingdom. 544 p. Research Foundation, Lunenburg, Massachusetts,

Spawls S, Howell K, Hinkel H, Menegon M. 2018. USA and Turtle Conservancy, Ojai, California, USA.

292 p.

Amphib. Reptile Conserv.

Flora Ihlow is a German herpetologist (Dr. rer. nat.) presently working at the Senckenberg Natural

History Collections, Dresden, Germany. For the past 10 years, Flora’s research has mainly focused

on the herpetofauna of Southeast Asia, in particular on the ecology, systematics, and distribution of

chelonians. She has published numerous scientific papers on these topics. After graduating from the

Rheinische Friedrich-Wilhelms-Universitat (Bonn, Germany), Flora joined the phylogeography group

of Senckenberg Dresden in 2017 as a post doc to study the systematics and distribution of chelonians

from southern Africa. Flora is a member of the IUCN/SSC Tortoise and Freshwater Turtle Specialist

Group (TFTSG).

Harith Farooq has been a Ph.D. student at the University of Aveiro, Portugal, and the University of

Gothenburg, Sweden, since 2016. Harith is supported by the WCS Christensen Conservation Leaders

Scholarship, the World Wildlife Foundation — Education for Nature Scholarship, and the Fundac¢éo

para a Ciéncia e Tecnologia. His main interests are biogeography and conservation, especially in

amphibians and reptiles. Harith has been inventorying these groups across Mozambique since 2011,

resulting in the publication of numerous species descriptions and range extensions. Before embarking

on his Ph.D., Harith worked at the Lurio University, Mozambique, for six years lecturing biological

sciences and publishing articles on science communication and environmental education.

Vaclav Gvozdik is a herpetologist based at the Institute of Vertebrate Biology of the Czech Academy

of Sciences. Vaclav is interested in the phylogeography, diversity, and evolution of amphibians and

reptiles of the Western Palearctic and sub-Saharan Africa. In Africa, he has mostly worked in lowland

and montane rainforests, and in recent years mainly in the rainforests of the Congo Basin. While

Vaclav has been studying various groups of herpetofauna, anurans have been his main focal group.

Margaretha D. Hofmeyr is Professor Emeritus at the Biodiversity and Conservation Biology

Department, University of the Western Cape, South Africa. Margaretha is an ecophysiologist by

training and first studied large ungulates before switching to chelonians. Her ecophysiological studies

revealed that South African tortoises have many unique characteristics, which stimulated her interest

in their genetic diversity and systematics. Margaretha has published extensively on the ecology

and phylogeography of sub-Saharan tortoises and turtles, and she is closely involved in several

conservation projects for threatened tortoises. This work resulted in her being awarded the 2015 Sabin

Turtle Conservation Prize. Margaretha is a member, and Regional Vice-Chair for Africa, of the IUCN/

SSC TFTSG and she coordinated the 2014 and 2018 Red List Assessments for South African tortoises

and freshwater turtles.

Werner Conradie holds a Masters in Environmental Science (M. Env. Sc.) and has 12 years of

experience with the southern African herpetofauna, with his main research interests focusing on the

taxonomy, conservation, and ecology of amphibians and reptiles. Werner has published numerous

principal and collaborative scientific papers, and has served on a number of conservation and

scientific panels, including the Southern African Reptile and Amphibian Relisting Committees. He

has undertaken research expeditions to many African countries including Angola, Botswana, Lesotho,

Malawi, Mozambique, Namibia, South Africa, Zambia, and Zimbabwe. Werner is currently the

Curator of Herpetology at the Port Elizabeth Museum (Bayworld), South Africa.

66 November 2019 | Volume 13 | Number 2 | e195

Amphib. Reptile Conserv.

lhlow et al.

Patrick D. Campbell holds a B.Sc. in Biological Sciences with 33 years working experience in the

department of Life Sciences (Zoology) at the Natural History Museum, London, NHM (UK). Patrick

has travelled the world on official duty as collector, diver, science officer, surveyor, and speaker at

various conferences, as far afield as China, Brazil, Thailand, Kenya, French Guiana, Ecuador, the

United States, Spain, and Milos to name but a few. He has published nearly 50, mostly collaborative,

papers on a variety of topics involving a number of different lower vertebrate species but most recently,

and primarily, on the taxonomy, osteology, and conservation of reptiles. Patrick is currently the Senior

Curator of Reptiles at the NHM (UK).

James Harvey works as an independent herpetologist, ecological researcher, and consultant, living in

South Africa. He holds degrees in Zoology, Hydrology, and Environmental Management, and has 16

years’ experience working with faunal biodiversity. James has performed ecological fieldwork widely,

primarily within Africa, in such countries as South Africa, Botswana, Zimbabwe, Angola, Malawi,

Mozambique, Kenya, Mali, Madagascar, Vietnam, and the Democratic Republic of the Congo. His

interests are diverse but center on the taxonomy, ecology, and conservation of herpetofauna and

other biodiverse groups. James has contributed to conservation assessments, workshops, and Red

Data publications on reptiles, amphibians, mammals, and plants for the southern and eastern African

regions. He regularly attends herpetological conferences, and has published numerous scientific papers

and was a contributing author on many more.

Luke Verburgt is a specialist consulting herpetologist working throughout A frica, with his professional

and scientific research experience extending over 16 years. Luke has published 18 internationally

recognized scientific papers to date on topics including herpetology, evolutionary biology, ecological

physiology, and animal behavior. His professional career covers biodiversity-related work on projects

throughout Africa and its islands (Angola, Botswana, Cameroon, Céte d'Ivoire, Guinea, Lesotho,

Liberia, Madagascar, Malawi, Mali, Marion Island, Mozambique, Namibia, South Africa, Uganda,

and Zimbabwe). Luke currently co-owns and co-directs the Enviro-Insight consultancy (http://www.

enviro-insight.co.za) where he fulfills roles as Director, senior ecological specialist, project manager,

software developer, and GIS specialist.

Uwe Fritz is the head of the Museum of Zoology, Senckenberg Natural History Collections at Dresden,

Germany, and Extraordinary Professor for Zoology at the University of Leipzig, Germany. Uwe has

worked for many years on the taxonomy, systematics, and phylogeography of turtles and tortoises,

and has also studied to a lesser extent snakes and lizards. He is particularly interested in hybridization

patterns and gene flow in contact zones of distinct taxa. Uwe has authored or co-authored numerous

scientific articles, mainly in herpetology, and has also edited various proceedings and books, among

them the two turtle volumes of the Handbook of Amphibians and Reptiles of Europe.

67 November 2019 | Volume 13 | Number 2 | e195

Official journal website:

amphibian-reptile-conservation.org

Amphibian & Reptile Conservation

13(2) [Special Section]: 68-81 (e197).

Value of forest remnants for montane amphibians on the

livestock grazed Mount Mbam, Cameroon

12.*Arnaud M. Tchassem Fokoua, ‘Legrand Nono Gonwouo, *Joseph L. Tamesse,

and *4Thomas M. Doherty-Bone

‘Laboratory of Zoology, Faculty of Sciences, University of Yaoundé I, P.O. Box 812, Yaoundé, CAMEROON ?*Higher Teacher Training College,

Department of Biological Science, Laboratory of Zoology, Yaoundé, CAMEROON 3?Conservation Programmes, Royal Zoological Society of

Scotland, Edinburgh, UNITED KINGDOM ‘Department of Life Sciences, Natural History Museum, London, UNITED KINGDOM

Abstract.—Habitat loss and degradation are the primary threats to biodiversity, especially for amphibians. In

the Highlands of Cameroon, knowledge on the impacts of different forms of habitat loss, such as livestock

management, is restricted to anecdotal reports. This study investigated the impact of forest fragmentation,

driven primarily by livestock grazing, on the amphibian assemblage on Mount Mbam, West Region, Cameroon.

Stratified, multi-season surveys over two years recorded the abundance and community composition of

anuran species. Based on the revised inventory of amphibians the proportion of threatened species on Mount

Mbam was calculated at 23.52%. A small population of Phrynobatrachus steindachneri was found to occur

despite having completely disappeared on other mountains in its distribution range. One species known to the

mountain, Cardioglossa schioetzi, was not found during the surveys. The remaining forest patches were found

to be significant habitat for several species endemic to the mountains of Cameroon-Nigeria. The savanna,

likely expanded by livestock grazing, held numerous reed frog species that likely benefit from forest loss,

especially in low- to mid-range elevations. The observed relationship between land-use and amphibians on

this mountain indicates that the ongoing conversion of forest to pasture threatens remaining montane endemic

anuran species, with conservation planning and action now necessary.

Keywords. Africa, endemic, frog, grassland, habitat fragmentation, habitat loss

Citation: Tchassem Fokoua AM, Gonwouo LN, Tamesse JL, Doherty-Bone TM. 2019. Value of forest remnants for montane amphibians on the

livestock grazed Mount Mbam, Cameroon. Amphibian & Reptile Conservation 13(2) [Special Section]: 68-81 (e197).

[Attribution 4.0 International (CC BY 4.0): https://creativecommons.org/licenses/by/4.0/], which permits unrestricted use, distribution, and reproduction

in any medium, provided the original author and source are credited. The official and authorized publication credit sources, which will be duly enforced,

are as follows: official journal title Amphibian & Reptile Conservation; official journal website: amphibian-reptile-conservation.org.

Received: 9 March 2019; Accepted: 15 August 2019; Published: 7 November 2019

Introduction

Vertebrate animal taxa are disappearing worldwide at

high rates; especially amphibians, a group which has

a high proportion of threatened species (Stuart et al.

2004; Beebee and Griffiths 2005). The five main factors

believed to be driving global amphibian declines are the

introduction of alien species, habitat alteration, over-

exploitation, global change, and infectious diseases

(Collins and Storfer 2003). The most severe stressor is

physical habitat degradation and destruction (Noss et al.

1997; Stuart et al. 2006). These factors have been found

in combination with enigmatic amphibian declines,

occurring particularly at localities above 400 m asl

in the Americas, Europe, and Australia (Pounds et al.

2006; Vences and Kolher 2008). However, studies on

amphibians in African mountains are limited to a handful

of sites, although recent findings have shown declines

in the Cameroonian mountains that are either enigmatic

(possibly associated with chytridiomycosis) or linked to

Correspondence. * arnaudtchassem@yahoo.fr

Amphib. Reptile Conserv.

habitat loss (Hirschfeld et al. 2016; Tchassem et al., in

press).

Cameroon is one of the most diverse countries in

terms of amphibian species, and it is home to about 4%

of the world’s known species of frogs (IUCN 2017). The

Bamenda Highlands form the northern portion of the

Highlands of Cameroon that includes Mount Mbam. Some

species known to occur in Mount Mbam are only known

from one or a few mountains, including some species

classified as Vulnerable or Endangered in the IUCN Red

List. This mountain has been studied less extensively

than many others, but its study should be a priority given

the community level amphibian declines in neighboring

mountains (Hirschfeld et al. 2016; Tchassem et al., in

press), where the causes remain uncertain as to whether

they are habitat driven or the result to other factors such as

the chtyrid fungus (Batrochochytrium dendrobatidis | Bd).

The impacts of environmental change on the Cameroon

mountains are still unclear, especially regarding habitat

loss, due to limited studies across Sub-Saharan Africa in

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Tchassem Fokoua et al.

general (Buckley and Jetz 2007).

Deforestation rates are exceptionally high across

West and Central Africa (Hansen et al. 2013). In the

biodiverse highlands of Cameroon and Nigeria (Bergl

et al. 2007), forest loss is driven by a combination of

clearance for cultivation, overexploitation of wood

fuel, and incursions by livestock (Chapman et al.

2004). The impact of this loss of forest on amphibians

has so far been assumed to be the loss of montane

endemic species, but the specific factors driving the

declines that are associated with forest loss have

not been determined. This uncertainty includes the

potentially differential impacts of livestock grazing

versus cultivation, for example, with potential

exacerbation by agrochemicals (Tchassem et al.,

in press). This scenario is especially applicable to

the degradation of natural habitats in the Bamenda

Highlands of Cameroon, that include Mounts Oku,

Bamboutos, Lefo, and Mbam, which share similar

montane endemic species (Cheek et al. 2000; Doherty-

Bone and Gvozdik 2017). Montane forest cover

has decreased dramatically during the 20" and 21

centuries; with remaining forests becoming highly

fragmented (Abbot et al. 2001; Doherty-Bone and

Gvozdik 2017). Agrochemical pollution and grazing

have not been systematically studied in this area, and

since they often occur simultaneously, differentiating

the impacts of each stressor is difficult (Tchassem et

al., In press).

Mount Mbam has historically been appraised for

amphibian diversity, with several montane endemic

species identified in the 1970s, such as Perret’s Egg Frog

(Leptodatylodon perreti) and Steindachner’s Puddle

Frog (Phrynobatrachus steindachneri) [Amiet 1971,

1973, 1976]. Despite this work, the mountain has no

official protection, and deforestation remains unchecked.

Unlike other mountains in the Bamenda Highlands,

Mount Mbam does not have considerable cultivation of

crops in contrast to its widespread livestock grazing. This

study focuses on Mount Mbam to assess the status of the

amphibian community and threats from encroachment

by livestock grazing. Grazed areas and remnant forests

were surveyed, incorporating an elevational gradient,

to determine: (1) whether the diversity of amphibians

has been altered compared to historical records; and (2)

which amphibian species are excluded from the grazed

areas compared to the gallery forest.

Materials and Methods

Study area. Mount Mbam (sometimes known as the

Mbam Hill Forest), is a calcareous massif located near the

town of Foumban in the West Region of Cameroon (Fig.

1, 05°57°N, 10°44’E). It rises from the savanna plains

at 1,000 m to a summit of 2,100 m asl. The vegetation

of the mountain 1s transitional between montane savanna

grassland mixed with patches of gallery forest (Fotso

Amphib. Reptile Conserv.

et al. 2001; Fig. 2). The forest patches are dominated

by Albizia gummifera, Polyscias fulva, and Schefflera

mannii (Fotso et al. 2001). Human communities living

around and using the mountain include the Banso’o,

Haoussa, and Fulani peoples. Livestock grazing is

predominately practiced by the Fulani community, who

are semi-nomadic, and concentrated at higher elevations

where they live for nine months during the year, from

April to December. During January to March (the dry

season), these cattle herders move down the mountain to

other pastures at lower altitudes.

Study design. To ensure a representative appraisal

of diversity, the Mount Mbam amphibian fauna was

surveyed over a period of two years, between 2014 and

2016. Field surveys were deployed, with all sample

sites visited at least three times. During diurnal and

nocturnal visual encounter surveys (VES) with a total

duration of 96 surveyor-hours across each land use type

were used to quantitatively sample fossorial, arboreal,

and water-associated amphibian species (Rodel and

Ernst 2004). Nocturnal acoustic encounter surveys were

also used to detect animals. Opportunistic sampling

was made for all taxa throughout the survey to enable

additions to the updated inventory. All sampling sites

were characterized with the following environmental

data: altitude and coordinates (recorded with a Garmin®

GPS exter 90), presence of potential breeding sites,

regime of disturbance, and vegetation structure. The

vegetation was determined at three strata (canopy cover,

shrubs, and understory). Surveys were stratified between

gallery forest (the only remnant forest on the mountain)

and grassland (primarily grazed by livestock). Surveys

involved searching of microhabitats, such as lifting rocks

and logs, peeling away bark from trees, moving fallen

debris, and inspecting tree stems during daytime (07:00-

12:00 h) and night time (19:00—00:00 h) along streams,

ponds, and the surrounding vegetation (Crump and Scott

1994; Rodel and Ernst 2004). Amphibians were captured

by hand, and then identified and released where they

were found. However, a subset of 1-3 individuals of

each species, and specimens difficult to identify in the

field, were euthanized using an overdose of MS-222 or

chlorobutanol solution and preserved in 75% ethanol.

These specimens have been deposited in the University

of Yaoundé I, Laboratory of Zoology. For species

identification, original descriptions and derived literature

were used (Perret 1966; Amiet 1977, 1980, 2012: Schiotz

1999).

Data Analysis. The proportion of threatened species

in the total amphibian species inventory for Mount

Mbam (both historical and contemporary species

observed) was calculated following Bohm et al. (2013).

All other analyses were performed with the statistical

package R version 3.3.2 (R Core Team 2016). Inventory

completeness was assessed using an incidence-based

November 2019 | Volume 13 | Number 2 | e197

Amphibians of Mount Mbam, Cameroon

a Ny ' } Le*

ty 1

jo —_ Summit iF

Ll 4S

q : if } RM nal .

i Wh j = c~\\ =

rn | ®) | oe wy

ag a ALA a

i | VJ it |

Li a

1 | | hy |

tr | 1

} 4 ry

i |e

| } a 6f i N

| J | .

| af | und i

| i

1.5 Km

a j* \ 7 7

fod -_ A Pasture

kJ 7

} 4 Swap area

e if

ig ae @ Forest patches

a8 a

@ Settlement

Fig. 1. Maps showing (top) the topography of the Bamenda Highlands, white circle showing Mount Mbam in the West Region of

Cameroon; and (bottom) the layout of sample sites on Mount Mbam.

estimator with the package BiodiversityR (Colwell and

Coddington 1994). To compare the amphibian community

composition between habitat types, PERMANOVA

analysis (formula: Adonis, library: vegan) was used to

test the significance of the Bray-Curtis dissimilarity

based on 999 permutations (Anderson 2001; Riemann

et al. 2017). Amphibian abundance data transformed

by square root were subjected to ordination analysis

using non-metric scale (NMDS) plots of Bray-Curtis

dissimilarity (formula: metaMDS, library: vegan) to view

the dispersion of similarities. The species contribution

to differences between different types of habitats was

evaluated by SIMPER analysis (Clarke 1993). The

influence of habitat (forest patches, pasture-grassland)

on species abundance was evaluated using Generalized

Linear Models (GLM, formula: glm, data family:

Amphib. Reptile Conserv.

poisson) [O’Hara and Kotze 2010]. Various parameters

such as season, year, and elevation were incorporated

into the GLMs to assess potential confounding factors

against the response variables (abundance of frogs).

GLMs were restricted to species for which a minimum

of five individuals were recorded in the surveys.

Information criterion analysis was then applied to the

GLMs, in which derived Akaike’s Information Criteria

were used to assess the best performing model based on

incorporation of these potentially competing explanatory

factors (Mazerolle 2006).

Results

The surveys revealed 17 anuran species of seven genera

among 225 individuals (Table 2). Based on these and

November 2019 | Volume 13 | Number 2 | e197

Tchassem Fokoua et al.

Bech nth es Ed re ae RR a 2 game a’ ae

Fig. 2. Montane habitats of amphibian species observed in recent surveys of Mount Mbam, West-Region, Cameroon: a) gallery

forest during the rainy season; b): gallery forest during the dry season after a bushfire; c) savanna area transformed by overgrazing;

and d): effects of bushfire started for pasture on the same site during the dry season.

historical records, the proportion of threatened species ==rheophilus, which shared the same breeding sites

on Mount Mbam is estimated at 23.52%. However, the (streams) [Table 3]. Leptopelis notatus was more

species accumulation curve over the survey periods did dominant in forests at lower elevations (1,307—1,340 m).

show a plateau, indicating the discovery of more species The only anurans of Phryobatrachidae on the mountain

with further searching is unlikely (Fig. 3). In comparison — were P. steindachneri restricted to forests at 1,400—1,800

to historical records, one species missing from the m (Tables 2—3). The only Hyperoliidae found in the

contemporary surveys was Cardioglossa schioetzi (Table forests were a singleton of Afrixalus aff. fulvovittatus (at

1). The Puddle Frog, Phrynobatrachus steindachneri, 1,342 m) and two individuals of Hyperolius ighettensis

was found, but only four sub-adult individuals were (at 1,342—1,684 m), though they likely spilled over from

observed over the two years. Species new to the historical the grassland areas.

inventory included the Rocket Frog Ptychadena Within derived savanna (i.e., savanna created by cattle

mascareniensis “D” (Zimkus et al. 2016) andthe Clawed grazing and fire) Hyperoliidae (43.55% of all individual

Frog Xenopus cf. eysoole (Fig. 4m). The latter species frogs) was the most dominant group, represented

was found as high as 1,600 m asl in stagnant water bodies — primarily below 1,500 m by HAyperolius concolor, H.

(including wells) that were heavily frequented by local balfouri, H. tuberculatus, H. igbettensis, H. nitidulus, H.

people and livestock. cinnamomeoventris, and A. aff. fulvovittatus (Tables 2-3).

Community structure (measured by Bray-Curtis — This family was followed by Pipidae (25.8% of individuals,

dissimilarity) between the two habitat types was _ represented by X. cf. eysoole only), Arthroleptidae (7%

significantly different (PERMANOVA: p = 0.01, Fig. — for LZ. notatus, L. nordequatorialis, as well as a minority

5). Most amphibian species were found in savanna (13 _ of five individual A. montanus), Ptychadenidae (5.8%,

species, 76.47% of individual frogs), with seven species represented by P mascareniensis “D” only), Bufonidae

(16% of individuals) found in forest (Table 2). Gallery = (2.2% for Sclerophrys maculata only), and Dicroglossidae

forests were dominated by species of Arthroleptidae, (2.2% for Hoplobatrachus occipitalis only). Four species

notably Astylosternus montanus, L. perreti, and A. occurred in both habitats: A. aff. fulvovittatus, A. montanus,

Amphib. Reptile Conserv. 71 November 2019 | Volume 13 | Number 2 | e197

Amphibians of Mount Mbam, Cameroon

Table 1. An updated amphibian species inventory for Mount Mbam, Cameroon.

Anuran taxa

Arthroleptidae

Astylosternus montanus Amiet, 1978

Astylosternus rheophilus Amiet, 1977

Cardioglossa schioetzi Amiet, 1982

Leptodactylodon perreti Amiet, 1971

Leptopelis nordequatorialis Perret, 1966

Leptopelis notatus (Peters, 1875)

Bufonidae

Sclerophrys maculata (Hallowell, 1854)

Hoplobatrachus occipitalis Gunther, 1858

Hyperolidae

Afrixalus aff. fulvovittatus Pickersgill, 2007

Hyperolius balfouri Werner, 1907

Hyperolius cf. cinnamomeoventris Bocage, 1866

Hyperolius igbettensis Mertens, 1940

Peters, 1875

Mocquard, 1897

Hyperolius nitidulus

Hyperolius tuberculatus

Pipidae

Xenopus cf. eysoole Evans et al. 2015

Phrynobatrachidae

Phrynobatrachus steindachneri Neiden, 1910

Ptychadenidae

Ptychadena mascareniensis “D” Zimkus et al. 2017

Species authority Global IUCN status

EC;

Le.

Endemicity References

CNH Amiet 1978; present study

CNH Amiet 1977; Hirschfeld et al. 2016; present

study

CNH Amiet 1982; Schietz 2004

BamH Amiet 1980; present study

CNH Perret 1966; Amiet 1971, 1974, 1980;

Gartshore 1986; present study

S-Sa Boulenger 1906; Nieden 1909; Goin 1961;

present study

PanAfr Hirschfeld et al. 2016; present study

S-Sa present study

S-Sa Perret 1976; present study

S-Sa Werner 1908; Scortecci 1943; Monard,

1951; present study

S-Sa Inger 1968; Largen and Dowsett-Lemaire

1991; Schiotz 1999; present study

S-Sa Schiotz 1963; present study

S-Sa Perret 1966; present study

S-Sa present study

present study

CNH Mertens 1968; Hirschfeld et al. 2016

S-Sa present study

Endemicity codes for species limited to: S-Sa - sub-Saharan Africa; C.W.A+Ng - Central and West African countries and Nigeria; BamH — just

the Bamenda Highlands of Cameroon that includes Mount Mbam; CNH: just the Bamenda Highlands of Cameroon and Nigeria.

H. igbettensis, and L. notatus.

Species represented by at least five individuals,

regardless of habitat type, varied with the strength of

the models in relation to habitat type. Species usually

associated with forest had better fitted models with

habitat (AAIC 0-7) and significant p-values, notably

Astylosternus sp. and L. perreti, as well as some savanna

species such as P. mascariensis “D,” H. nititdulus, and H.

tuberculatus (Table 4). Species associated with savanna

(Afrixalus, some Hyperolius sp., and S. maculata) and L.

notatus, however, did not show statistically significant or

well fitted models with habitat as a fixed variable (Table

4). Habitat* elevation and season* elevation both provided

the best fitted models (the lowest AIC) for A. rheophilus,

with a AAIC of seven from the inclusion of habitat

alone or from the inclusion of habitat*elevation* season.

Habitat is a major contributor for the best fitted

models for the species A. montanus (habitat*year), L.

notatus (habitat*elevation), and P mascareniensis “D”

(habitat*season) [Table 4]. Year of survey did have an

influence over the fit of the models, with the exceptions

of H. cinnamomeoventris, P. mascareniensis, and S.

Amphib. Reptile Conserv.

maculata, manifest by all three having fewer records for

the year 2016. For the remaining species, the lack of an

influence of year indicates population stability in this

two-year time period on the mountain.

Discussion

This study quantitatively assessed the status and habitat

use of amphibians on Mount Mbam, Cameroon, in rela-

tion to land use. The species inventory of the amphibians

of the mountain was updated, revealing little additional

diversity recorded for the Mbam massif so far, beyond

more lowland-adapted species. The stratified survey en-

abled better understanding of the habitat requirements of

numerous amphibian species, especially montane species

with restricted ranges. Species composition varied con-

siderably between montane savanna and forest habitats.

With a higher elevation, the forest was generally found

to have fewer species, but with considerably more nu-

merous montane endemic species. There were clear 1n-

stances of spillover from one habitat to another, typified

by the occurrences of a minority of species in one habitat

compared to the habitat in which they are numerically

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Tchassem Fokoua et al.

Number of species

30 40

pecee eens see e ee ee EPO P EERE EEE ED

eeaseeeea ear eeee

saee

weeeet

eet

eaaaee

anor

wat

aenet

ate

Savanna

— Forest

60 70 80 90 100

Samples

Fig. 3. Species accumulation curves of Mount Mbam by land use based on contemporary records.

dominant.

Compared to nearby mountains such as Oku

or Bamboutos, Mount Mbam has a more diverse

community of hyperoliids, likely due to a greater area

of the mountain consisting of lower elevations. This

is in contrast to mountains such as Bamboutos, where

agrochemicals are widely used. It is notable that the

genera Arthroleptis and Kassina were not observed

in either historical or contemporary surveys, and the

explanation for their absence is unclear. Montane

endemic species known to Mounts Lefo, Oku, and

Bamboutos, including Cardioglossa oreas, C. pulchra,

and Astylosternus ranoides, were not found on Mbam.

This is despite other species such as Leptodactylodon

perreti and A. rheophilus occurring on this mountain,

and the fact that these absent species do have elevational

ranges corresponding to that of Mt. Mbam. This suggests

the possibilities that they are either: (1) locally extinct

through human land use practices or climate change that

pre-date the first surveys of the mountain in the 1960s, or

(ii) that the mountain is too small and low in elevation for

viable populations of the higher montane endemic frog

species to persist.

Despite its disappearances on Mount Oku in the

North West Region and Mount Bamboutos in the West

Region (Doherty-Bone and Gvozdik 2017; Tchassem

et al., in press), P steindachneri was observed during

the recent field surveys. This taxon is part of a species

complex that could possibly also include P. jimzimkusi

(Zimkus and Gvozdik 2013). Cardioglossa_ schioetzi

was not observed in this study, and it was also not found

on Mount Oku in recent years, as with certain other

species of Cardioglossa, Werneria, and Phrynobatrachus

(Hirschfeld et al. 2016; Doherty-Bone and GvoZzdik

2017; Tchassem et al., in press). The causes of these

disappearances remain unknown, but the declines on

Mount Manengouba and Mount Oku have coincided

with an increase in the prevalence of amphibian chytrid

fungus (Batrachochytrium dendrobatidis, Bd), indicating

that disease could be a factor (Hirschfeld et al. 2016).

Amphib. Reptile Conserv.

The role of climate change in these declines also remains

unclear and represents a research gap that requires urgent

attention (Doherty-Bone and GvoZdik 2017; Tchassem et

al., in press). On Mount Mbam, further research should

include investigating the role of fire, which could be

a factor, not just from its use by livestock herders but

Table 2. Summary of amphibian species (total per habitat type

across all survey techniques) encountered during the present

study (2014—2016).

Pasture-

grassland

Afrixalus aff. fulvovittatus Pe 1

Gallery

Species forest

—

Astylosternus montanus

Astylosternus rheophilus

Hoplobatrachus occipitalis

Hyperolius balfouri

Hyperolius tuberculatus

Hyperolius concolor

Hyperolius igbettensis Gi

Hyperolius nitidulus

Hyperolius cinnamomeoventris 7

Leptodactylodon perreti 5

Leptopelis nordequatorialis 2

Leptopelis notatus

Phrynobatrachus steindachneri

Sclerophrys maculata f tl

Ptychadena mascareniensis “D” 13:

Xenopus cf. eysoole 8

189

23.63

November 2019 | Volume 13 | Number 2 | e197

Total 3

Ls 0

Dee Se

Species richness 7

0.80

2.44

4.50

Species evenness

Shannon’s D

Mean number of specimens per sampling

event

Amphibians of Mount Mbam, Cameroon

' Y= | a a : ie

Fig. 4. Montane endemic amphibian species observed in recent surveys of Mount Mbam, West-Region, Cameroon. a) Astylosternus

rheophilus, b) Astylosternus montanus, ¢) Afrixalus aff. fulvovittatus, d) Hyperolius balfouri, e) Hyperolius igbettensis, f)

Hyperolius nitidulus, g) Hyperolius concolor, h) Hyperolius cinnamomeoventris, i) Hyperolius tuberculatus, j) Leptopelis

nordequatorialis, k) Leptopelis boulengeri, 1) Phrynobatrachus steindachneri, m) Xenopus cf. eysoole, n) Hoplobatrachus

occipitalis, and 0) Sclerophrys maculata.

Amphib. Reptile Conserv. 74 November 2019 | Volume 13 | Number 2 | e197

Tchassem Fokoua et al.

Stress = 0.00005

Permanova: p<0,01

NMDS2

(e)

Savanna Area

Forest patches

-2

-4

-5 0 5

NMDS1

Fig. 5. Non-metric dimensional scaling plot of amphibian

community structure divided by land use type on Mount Mbam

based on visual encounter surveys with equal effort for each land

use. The PERMANOVA p-value is shown in the top right corner.

also its influence by climate change. Some species were

possibly overlooked during this study, possibly due to

variance in the detectability of rare and/or cryptic species

(Mackenzie et al. 2005; Megson et al. 2009); thus,

further efforts to continue this study would benefit from

complementary survey methods such as pitfall traps and

continuous audio recordings. However, it is also possible

that the species pool for Mount Mbam has simply been

degraded historically.

The presence of livestock grazing on the slopes of

Mount Mbam has likely modified the landscape structure

of habitats to a great extent, such as through loss of

forest from grazing, fires, and trampling by livestock

(Carte and John 2002). The remaining forest on Mount

Mbam is now reduced to patches along streams and close

to the summit where access is difficult for livestock.

The historical loss of forest is likely to have negatively

affected populations of forest-dependent species, while

potentially benefitting tolerant, savanna-adapted species.

While the forests hosted fewer species than savanna,

those species are montane endemics of conservation

concern, while the greater species richness of the savanna

consists of species with broader, lowland ranges. Most of

the amphibian species were significantly influenced by

elevation, habitat, and season. This indicates that despite

more savanna becoming available, the colonization by

lowland species may remain limited.

Planning and implementing effective strategies to

control habitat disturbance and encourage recovery

on this mountain could be required to stop further loss

of montane endemic species in the long term. Future

studies should investigate the precise impacts of bushfire,

including influences on nutrient cycling, water quality,

predation risk to anurans, and reproductive success. The

conservation needs of Mount Mbam are similar to those

of Mount Bamboutos, but are driven more by livestock

than the cultivation of crops. As with Mount Bamboutos,

Mount Mbam has neither official protection nor

conservation action, and the exploitation of its resources

is unregulated. Several measures could be implemented

to reduce the rate of forest loss on Mount Mbam. Steps

such as raising environmental awareness, conducting

educational seminars, and preparing educational

materials for the locals, would certainly have a positive

Table 3. Similarity percentage (SIMPER) analysis showing importance of dissimilarity for various amphibian species for habitat

type based on visual encounter surveys of Mount Mbam.

Mean number of individuals per survey

Taxon Forest Pasture-grassland eet mes te wana

Xenopus cf. eysoole 0+0 0.64 + 1.24 0.17 0.17

Astylosternus montanus 0.47 + 0.86 0.06 + 0.23 0.14 0.32

Leptopelis notatus 0.17+0.42 0.07 + 0.39 0.08 0.39

Hyperolius balfouri 0+0 0.23 + 0.75 0.07 0.46

FHyperolius concolor 0+0 0.24 + 0.96 0.06 0.58

Afrixalus aff. fulvovittatus 0.03 + 0.18 0.17 + 0.60 0.06 0.52

Astylosternus rheophilus 0.17+ 0.46 0+0 0.05 0.69

Hyperolius igbettensis 0.07 + 0.25 0.08 + 0.37 0.05 0.79

Hyperolius tuberculatus 0+0 0.17 + 0.64 0.05 0.89

Phrynobatrachus steindachneri 0.13 + 0.35 0+0 0.05 0.84

Ptychadena mascareniensis 0+0 0.14+0.49 0.05 0.63

FHyperolius cinnamomeoventris 0+0 0.08 + 0.37 0.03 0.95

Hyperolius nitidulus 0+0 0.09 + 0.41 0.03 0.92

Sclerophrys maculata 0+0 0.06 + 0.23 0.03 0.97

Hoplobatrachus occipitalis 0+0 0.06 + 0.38 0.02 0.99

Leptopelis nordequatorialis 0+0 0.02 + 0.15 0.01 1

Amphib. Reptile Conserv. 75 November 2019 | Volume 13 | Number 2 | e197

Amphibians of Mount Mbam, Cameroon

Table 4. Generalized linear models comparing parameters which influence the abundance of amphibian species on Mount Mbam.

Species

Afrixalus aff. fulvovittatus

Astylosternus montanus

Astylosternus rheophilus

Hoplobatrachus occipitalis

Hyperolius balfouri

Amphib. Reptile Conserv.

Model parameters

Habitat* Elevation* Season

Habitat* Elevation

Habitat* Season

Season* Elevation

Habitat* Year

Habitat

Elevation

Season

Year

Habitat* Elevation* Season

Habitat* Elevation

Habitat* Season

Season* Elevation

Habitat* Year

Habitat

Elevation

Season

Year

Habitat* Elevation* Season

Habitat* Elevation

Habitat* Season

Season* Elevation

Habitat* Year

Habitat

Elevation

Season

Year

Habitat* Elevation* Season

Habitat* Elevation

Habitat* Season

Season* Elevation

Habitat* Year

Habitat

Elevation

Season

Year

Habitat* Elevation* Season

Habitat* Elevation

Habitat* Season

Season* Elevation

Habitat* Year

Habitat

Elevation

112

116

118

112

116

118

112

116

118

112

116

118

112

116

118

Residual

deviance

41.48

63552

62.95

47.88

77.78

84.83

74.05

81.87

88.35

51.96

a9: 92

56.69

64.91

53.47

64.11

TT9

68.49

81.52

9.76

20.00

992

20.51

20.69

9:93

30.50

31.11

34.40

35.47

38.26

34.65

29.11

38.268

35.64

41.05

204.44

68.50

90.26

76.22

69.92

99.10

99.26

OV 2

November 2019 | Volume 13 | Number 2 | e197

p-value

0.99

<0.01

<0.001

0.01

0.05

<0.001

<0.01

0.53

0.99

0.06

0.99

0.90

<0.001

<0.01

<0.001

0.12

AIC

269.90

108

123

108

102

131

128

119

AAIC

Tchassem Fokoua et al.

Table 4 (Cont.). Generalized linear models comparing parameters which influence the abundance of amphibian species on Mount Mbam.

Species Model parameters df ae p-value AIC AAIC

Season 118 94.31 <0.001 123 21

Year 118 111.34 0.94 139 36

Hyperolius cinnamomeoventris _ Habitat*Elevation* Season 112 39.44 1.00 65 19

Habitat* Elevation 116 42.10 0.99 60 14

Habitat* Season 116 44.07 1.00 62 16

Season* Elevation 116 40.14 0.13 58 12

Habitat* Year 116 31.24 1.00 49 3

Habitat 118 44.07 0.04 58 12

Elevation 118 42.49 0.02 56 10

Season 118 47.83 0.60 62 16

Year 118 32.36 <0.001 46 0)

Hyperolius concolor Habitat* Elevation* Season 112 82.19 0.99 119 0

Habitat* Elevation 116 103.88 0.99 132 13

Habitat* Season 116 108.83 0.99 137 19

Season* Elevation 116 100.95 <0.001 129 10

Habitat* Year 116 116.84 0.99 145 26

Habitat 118 11732. <0.001 142 2

Elevation 118 129.98 0.97 154 35

Season 118 125.50 0.03 150 31

Year 118 129.81 0.68 154 ae

Hyperolius igbettensis Habitat* Elevation* Season 112 044.07 0.19 74 2

Habitat* Elevation 116 53.05 0.36 7 3

Habitat* Season 116 52.12 0.13 74 2

Season* Elevation 116 53.54 0.46 75 3

Habitat* Year 116 54.83 0.81 76 4

Habitat 118 54.91 0.84 73 1

Elevation 118 54.30 0.42 72 0)

Season 118 54.46 0.49 Te. 0)

Year 118 54.93 0.91 73 1

Hyperolius nitidulus Habitat* Elevation* Season 112 43.35 1.00 71 i

Habitat* Elevation 116 48.10 0.99 68 4

Habitat* Season 116 4471 1.00 64 0

Season* Elevation 116 46.39 0.30 66 2

Habitat* Year 116 46.23 0.99 65 1

Habitat 118 48.10 0.03 64 0)

Elevation 118 50.29 0x12. 66 2

Season 118 50.85 0.17 66 3

Year 118 49.48 0.07 65 1

Hyperolius tuberculatus Habitat* Elevation* Season 112 73.36 0.99 106 7

Habitat* Elevation 116 79.67 0.99 104 >

Habitat* Season 116 75.91 0.99 100 1

Season* Elevation 116 74.91 0.33 99 0

Habitat* Year 116 81.60 0.99 106 7

Amphib. Reptile Conserv. 77 November 2019 | Volume 13 | Number 2 | e197

Amphibians of Mount Mbam, Cameroon

Table 4 (Cont.). Generalized linear models comparing parameters which influence the abundance of amphibian species on Mount Mbam.

Residual

Species Model parameters df aur ate p-value AIC AAIC

Habitat 118 81.85 <0. O1 103 4

Elevation 118 80.99 <0.01 101 2

Season 118 87.32 <0. O1 108

Year 118 90.41 0.79 111 | ie.

Leptodactylodon perreti Habitat* Elevation* Season 112 14.52 0.99 39 1]

Habitat* Elevation 116 14.54 0.99 31 3

Habitat* Season 116 20.00 1.00 aE 9

Season* Elevation 116 14.99 0.99 32 4

Habitat* Year 116 20.51 0.99 Bd 9

Habitat 118 20.69 <0.001 33 5

Elevation 118 15.07 <0.001 28 0)

Season 118 30.49 0.04 43 15

Year 118 34.12 0.51 46 18

Leptopelis notatus Habitat* Elevation* Season 112 31.79 0.99 65 14

Habitat* Elevation 116 43.04 <0.01 69 0

Habitat* Season 116 48.74 0.01 74 5

Season* Elevation 116 48.84 <0.01 74 5

Habitat* Year 116 58.95 0.85 84 14

Habitat 118 59.78 0.14 81 12

Elevation 118 61.16 0.38 83 12

Season 118 59.86 0.15 81 12

Year 118 60.98 0.33 82 13

Ptychadena mascareniensis “D” Habitat* Elevation* Season 112 52.10 1.00 88 4

Habitat* Elevation 116 61.89 0.99 90 6

Habitat* Season 116 55.95 0.99 84 0)

Season* Elevation 116 58.16 0.28 86 2

Habitat* Year 116 57.98 0.99 86 2

Habitat 118 62.44 <0.01 87 3

Elevation 118 67.71 0.14 90 6

Season 118 59.56 <0.01 84 0

Year 118 156 <0.001 90 6

Sclerophrys maculata Habitat* Elevation* Season 112 27.81 1.00 54 20

Habitat* Elevation 116 28.90 1.00 5] 23

Habitat* Season 116 28.27 0.99 46 12

Season* Elevation 116 28.49 0.33 46 12

Habitat* Year 116 19.74 0.99 38

Habitat 118 28.90 0.09 43

Elevation 118 30.34 0.23 44 10

Season 118 30.30 0.22 44 10

Year 118 20.54 <0.001 34 0

Xenopus cf. eysoole Habitat* Elevation* Season* Year -] 0 1.00 148 103

Habitat* Elevation* Season 112 151.00 1.00 228 183

Habitat* Elevation 116 172.00 0.99 241 196

Amphib. Reptile Conserv. 78 November 2019 | Volume 13 | Number 2 | e197

Tchassem Fokoua et al.

Table 4 (Cont.). Generalized linear models comparing parameters which influence the abundance of amphibian species on Mount Mbam.

Species Model parameters

Habitat* Season

Season* Elevation

Habitat* Year

Habitat

Elevation

Season

Year

effect on changing negative attitudes. The creation of a

protected area for these habitats, with strong involvement

of the local people, would be a plausible strategy. The

proportion of endangered species of the Mount Mbam

is very low, but this does not in any way diminish

its importance for the conservation of the endemic

amphibian species, whereas the differences observed at

low and high altitudes with regard to species composition

and habitat type should make this site a national or sub-

regional conservation priority.

Acknowledgments.—The authors express their gratitude

to the Cameroon Ministry of Scientific Research and

Innovation and the Ministry of Forestry and Wildlife

for authorizing our fieldwork (permit number 629PRS/

MINFOF/SG/DFAP/SDVEEF/SC). We especially thank

Mark Wilkinson, Natural History Museum, London,

United Kingdom for assistance. We thank the Rufford

Small Grant (RSG) for funding this work. In addition, we

wish to acknowledge the village chiefs and community

elders who permitted the work on their land, and thank

field assistants Mballa Moussa and Daho. The authors

also thank the reviewers for helping to improve this

manuscript.

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Amphib. Reptile Conserv.

Tchassem Fokoua et al.

Arnaud Marius Tchassem Fokoua is currently a third year Ph.D. candidate at the University

of Yaoundé I, Cameroon. Arnaud received his Bachelor’s degree at the same university,

completing multiple research projects in the domain of herpetology. His research focuses

on the community ecology and conservation of African amphibians, especially aiming

to understand how altitude and anthropogenic activities influence amphibian community

composition.

Legrand Gonwouo Nono (Ph.D.) is a conservation herpetologist from University of Yaoundé

I with experience in central African reptiles and amphibians. Dr. Gonwouo Nono has long-

term experience studying the amphibians and reptiles of Cameroon, particularly in the fields

of biodiversity, taxonomy, and ecology. His recent research interest focuses on the responses

of biodiversity to environmental change, with particular interest in the distributions and

dynamics of endemic species near their geographic range margins.

Joseph Lebel Tamesse is a professor at the Higher Teacher Training College in Cameroon.

Joseph Lebel has a great deal of experience in the study of the psyllids of Cameroon,

particularly in the fields of biodiversity, taxonomy, and ecology. His current studies integrate

multiple fields, and he also supervises several Ph.D. students with research which focuses

on the ecology of millipedes, amphibians, and freshwater crabs. He has co-authored over 50

scientific papers.

Thomas Doherty-Bone is an ecologist researching conservation biology, management, and

herpetology. Thomas is associated with the Royal Zoological Society of Scotland and the

Natural History Museum, London, and holds a Ph.D. in freshwater ecology and invasive alien

species from the University of Leeds, United Kingdom. His research has focused specifically

on montane amphibian ecology and conservation in Cameroon for over 12 years, including

the occurrence of amphibian chytrid fungus and the conservation of Lake Oku and its endemic

clawed frog (Xenopus longipes).

81 November 2019 | Volume 13 | Number 2 | e197

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Amphibian & Reptile Conservation

13(2) [Special Section]: 82-95 (e199).

urn:lsid:zoobank.org:pub:63D950B1-10B3-4EC1-B4E6-8558F5618DF6

Another Angolan Namib endemic species: a new

Nucras Gray, 1838 (Squamata: Lacertidae) from

south-western Angola

12William R. Branch, ***Werner Conradie, *°°Pedro Vaz Pinto, and ’*Krystal A. Tolley

'Port Elizabeth Museum, P.O. Box 13147, Humewood, Port Elizabeth 6013, SOUTH AFRICA *Department of Zoology, Nelson Mandela University,

Port Elizabeth 6031, SOUTH AFRICA °?School of Natural Resource Management, George Campus, Nelson Mandela University, George 6530,

SOUTH AFRICA ‘Fundagao Kissama, Rua 60 Casa 560, Lar do Patriota, Luanda, ANGOLA °CIBIO/InMBIO, Centro de Investigagdo em

Biodiversidade e Recursos Genéticos, Universidade do Porto, Campus de Vairdo, Vairdo, PORTUGAL °TwinLab CIBIO/ISCED, Instituto Superior

de Ciéncias da Educagdo da Huila, Rua Sarmento Rodrigues s/n, Lubango, ANGOLA ‘South African National Biodiversity Institute, Kirstenbosch

Research Centre, Private Bag X7, Claremont 7735, Cape Town, SOUTH AFRICA *Centre for Ecological Genomics and Wildlife Conservation,

Department of Zoology, University of Johannesburg, Auckland Park, 2000, Johannesburg, SOUTH AFRICA

Abstract—A new endemic Sandveld Lizard, genus Nucras, is described from south-western Angola.

Morphologically it resembles members of the Nucras tessellata group, but it is genetically separated and is sister

to the larger tessellata + lalandii group. Although the genus is generally very conservative morphologically,

the new species differs from other congeners in a combination of scalation, overall dorsal color pattern, and

geographic separation. The new species is known from fewer than 12 specimens collected over a period

spanning 120 years from arid south-western Angola. This brings the total number of species in the genus

to 12 and adds another species to the growing list of endemic species of the Namib region of Angola. This

new finding further reinforces the idea that this Kaokoveld Desert region is a key biodiversity area worthy of

conservation and long-term protection.

Keywords. Sandveld Lizard, taxonomy, Africa, endemism, Kaokoveld, biodiversity hotspot

Citation: Branch WR, Conradie W, Vaz Pinto P, Tolley KA. 2019. Another Angolan Namib endemic species: a new Nucras Gray, 1838 (Squamata:

Lacertidae) from south-western Angola. Amphibian & Reptile Conservation 13(2) [Special Section]: 82-95 (e199).

4.0 International (CC BY 4.0): https://creativecommons.org/licenses/by/4.0/], which permits unrestricted use, distribution, and reproduction in any

medium, provided the original author and source are credited. The official and authorized publication credit sources, which will be duly enforced, are

as follows: official journal title Amphibian & Reptile Conservation; official journal website: amphibian-reptile-conservation.org.

Received: 8 May 2019; Accepted: 8 August 2019; Published: 8 November 2019

Introduction At present, the family Lacertidae is represented in

Angola by 13 species in six genera; Heliobolus (one

species), Holaspis (one), Ichnotropis (three), Meroles

(three), Nucras (two), and Pedioplanis (three; see Marques

The recorded reptile diversity in Angola (278 species,

Marques et al. 2018; Branch et al. 2019) is significantly

lower than that of South Africa (407 species, Tolley et al.

2019), a nearby country of comparable size and habitat

diversity. This incongruity has been attributed to the lack of

recent faunal surveys and/or taxonomic revision of groups

in the country (Marques et al. 2018; Branch et al. 2019).

That this gap simply represents under-sampling of the

Angolan herpetofauna is evidenced by the recent discovery

of numerous new species, including lacertids of genus

Pedioplanis (Conradie et al. 2012), girdled lizards of genus

Cordylus (Stanley et al. 2016; Marques et al. 2019b), and a

new skink of genus Trachylepis (Marques et al. 2019a), as

well as several candidate new species of lacertids (Branch

and Tolley 2017), and geckos (Branch et al. 2017).

Correspondence. * werner@bayworld.co.za

Amphib. Reptile Conserv.

et al. 2018; Branch et al. 2019). The lacertid generic

diversity is comparable to that of other herpetologically

rich areas in sub-Saharan Africa, e.g., eight genera in

Tanzania, Kenya, and South Africa, and five in Namibia

(Branch 1998; Spawls et al. 2018). However, the lacertid

species diversity in Angola (13 species) is notably lower:

Kenya (15), Tanzania (16), Namibia (25), and South

Africa (28) [Branch 1998; Spawls et al. 2018; Branch et

al. 2019; Bauer et al. 2019].

The taxonomy of the lacertid genus Nucras Gray,

1838 is complicated by the relatively secretive habits

and conservative morphology of known species, and

this has confounded early attempts to resolve species

November 2019 | Volume 13 | Number 2 | e199

Branch et al.

boundaries and geographical distributions within the

genus. Currently, Nucras comprises eleven species that

are mainly restricted to southern Africa, with a northern

outlier (Nucras boulengeri Neumann, 1900) occurring

in East Africa, although there is a single, isolated record

from Isoka, northern Zambia (Haagner et al. 2000;

Spawls et al. 2018).

Taxonomy of the Western (or Striped) Sandveld

Lizard, Nucras tessellata, has proven to be particularly

problematic, as have the species boundaries within the N.

tessellata species complex. Broadley (1972) recognized

four subspecies (Nucras taeniolata taeniolata, Nucras

taeniolata ornata, Nucras tessellata tessellata, and

Nucras tessellata livida), as well as a number of

taxonomically unresolved non-specific forms, Le.,

Nucras tessellata tessellata var. “TV,” Nucras taeniolata

ornata var. holubi, and Nucras tessellata tessellata vat.

elegans. Broadley (1972) examined the morphology of

over 800 specimens and concluded that the dorsal color

pattern and the number of subdigital lamellae under the

4" toe are reliable taxonomic characters to differentiate

species within the N. tessel/lata complex. In recent years,

several subspecies and varieties were elevated to full

species, e.g., Nucras taeniolata, N. holubi, N. ornata

(Jacobsen 1989), and N. /ivida (Branch and Bauer 1995).

A number of historically problematic Angolan specimens

were considered to form part of the Nucras tessellata

(Smith, 1838) complex, best representing Nucras

tessellata tessellata var. “T.” However, Broadley (1972)

deferred making a decision on their taxonomic status

pending the collection of additional material. The only

other Angolan member of the genus, Nucras scalaris

Laurent, 1964, was described on the basis of material

from northern Angola and is not currently regarded to be

included in the N. tessellata complex.

Bocage (1895) was the first to record N. tessellata from

Angola, but noted only that (translated from the original

French): “Mr. Anchieta met this species at two different

locations, Maconjo and Caconda, from where he sent us

a few individuals. All of these individuals belong to the

variety taeniolata, separated from the typical form not

only by its coloration, with a back striped longitudinally

in white and blackish-brown, but is also slimmer.” He

provided no further details of the specimens, leaving out

information on scalation and size. Fortunately, the late

Donald G. Broadley visited the Museu Bocage Lisboa,

Portugal (currently Museu Nacional de Historia Natural

e da Ciéncia) in 1968, before the disastrous fire of 1978

destroyed its collections. Broadley was only able to locate

the three specimens from Maconjo listed by Bocage

(1895). Boulenger (1910) first assigned specimens from

Mocamedes (=Namibe) district to Nucras tessellata var.

taeniolata. In subsequent years, he referred the same

material as part of Nucras intertexta var. holubi under

a different color variation A and called this the most

“primitive form’ (Boulenger 1917, 1921). Monard (1937)

Amphib. Reptile Conserv.

recorded three additional juvenile specimens from

Kapelongo (= Capelongo) and reported that they exhibit

typical coloration of taeniolata, and thus assigned his

material to the N. tesse/lata complex. The most detailed

description to follow was a specimen collected from “km

34 de la route de Mocamedes a Sa da Bandeira” (= 34

km from Namibe on Lubango road) and documented by

Laurent (1964). All the above specimen data are pooled

in the summary tables of scalation in the revision of the

N. tessellata complex (Broadley 1972), and he concluded

that the Angolan material represents an undescribed

species.

During recent surveys in south-western Angola,

several individuals of Nucras were collected. This new

material is compared with historical material of the

species known from Angola and supplemented with

phylogenetic analyses to investigate their taxonomic

status, and to advance our understanding of the N.

tessellata complex.

Materials and Methods

Sampling and material examined. During a recent

expedition to south-western Angola, two Nucras

individuals were collected from Namibe Province (Fig.

1). Each specimen was collected as a voucher, fixed in

10% formalin and thereafter transferred to 70% ethanol

for long-term storage at the Port Elizabeth Museum

(PEM). Prior to fixation, a tissue sample was collected

and preserved in 99% ethanol. Material from the

following museums was examined (Table 1) by Donald

Broadley: Museu Bocage Lisboa, Portugal (MBL),

Museu Regional do Dundo, Dundo, Angola (MD), and

the British Museum (now Natural History Museum,

London) [NHML]. WRB examined material in the

Transvaal Museum (now Ditsong National Museum of

Natural History Northern Flagship Institute, Pretoria)

[TM], and re-examined and photographed the NHML

specimens. Photographs of Monard’s (1937) material

from the Musée d’Histoire Naturelle, La-Chaux-de-

Fond, Switzerland (MHNC, formerly LCFM) were made

available by Luis Ceriaco. The Angolan material was

further compared to other material housed in the PEM.

Morphological data. To quantify morphology for the

Species diagnoses, the following measurements were

recorded from each individual: snout-vent length (SVL):

tip of snout to anterior edge of cloaca; tail length (Tail):

tip of tail to posterior edge of cloaca; total length (TL):

combined SVL and tail length; head length (HL): from

anterior edge of occipital/parietal scale to tip of snout;

head width (HW): width of head (just behind eye); snout

length (SL): from anterior corner of eye to tip of snout;

eye length (EL): horizontal diameter of eye; ear-eye

length: from posterior corner of eye to anterior edge of

ear opening.

November 2019 | Volume 13 | Number 2 | e199

A new Nucras species from Angola

Legend

@ Nucras broadleyi sp. nov.

Altitude

Mi i00m

500 m

| 900m

1300 m

Ml 1700 m

150

Fig.1. Map showing distribution of Nucras broadleyi sp. nov. in Angola.

The following scalation details were recorded:

upperlabials (UL): in front of subocular and after

subocular; lowerlabials (LL), transverse rows of

ventrals, longitudinal ventral scale rows, supraciliars

(SC), granules between supraciliars (SC) and subocular,

number of subdigital lamellae below 4" toe, and number

of femoral pores. All counts were performed on both left

and right sides. The presence of interparietal and whether

it was in contact with occipital were also recorded.

Phylogenetic analyses. To place the two Nucras

individuals recently collected from Angola in a

phylogenetic context, one nuclear (RAG-1) and two

mitochondrial (ND4, 16S) genes were sequenced (Table

2). DNA was extracted using salt extraction (Aljanabi

and Martinez 1997), with PCR amplification, and cycle

sequencing following standard procedures. A 25 ul PCR

reaction included 3 ul of 1 mM dNTPs, 3 ul of 25 mM

MgCl, 0.2 ul of 10 pmol forward and reverse primers,

3 ul of buffer solution (20 mM Tris-HCI ~pH 8.0, 100

mM NaCl, 0.1 mM EDTA, 1 mM DTT), 0.1ul (0.5U)

Taq polymerase, and 1—2 ul of 25 ng/ul genomic DNA.

Thermal cycling was run with initial denaturation for 4

min at 94 °C followed by: 35 cycles with denaturation for

30 s at 94 °C, annealing for 40 s at 55—57 °C, extension

Amphib. Reptile Conserv.

for 40 s at 72 °C, and final extension for 4 min at 72 °C.

Primers used for amplification were ND4: ND4 (Forstner

et al. 1995) and Leul (Arévalo et al. 1994), 16S: L2510

and H3080 (Palumbi 1996); and RAG-1: RAGI-FO and

RAGI-R1 (Mayer and Pavlicev 2007). PCR products

were run on a 1% agarose gel and visualized under a UV

light to verify amplification. Amplicons were sequenced

directly using the forward primers at Macrogen

(Amsterdam, Netherlands). Sequences were edited and

aligned using Geneious software v4.7 (Kearse et al.

2012). New sequences have been deposited in GenBank

(Table 2). In addition, gene sequences for multiple

individuals of all Nucras species (except N. scalaris) and

sequences representing outgroup taxa were downloaded

from GenBank (Table 2).

A Bayesian analysis of 2,052 characters from the two

mitochondrial genes and one nuclear gene (ND4: 678

bp, 16S: 482 bp, RAG-1: 892 bp) was used to investigate

optimal tree space using MrBayes v3.2.2 (Huelsenbeck

and Ronquist 2001) at the CIPRES Science Gateway

(Miller et al. 2010). To determine which evolutionary

model best fit the data, j;Modeltest was initially run (Posada

2008). The AIC test specified the GIR+G model for both

mitochondrial markers and HYK+G for RAG-1. Therefore,

three unlinked data partitions were created, specifying six

November 2019 | Volume 13 | Number 2 | e199

Branch et al.

100

1.0

6.97

0.98

1.0

100

1.0

100

1.0

L, longicaudata

0.04 substitutions/site

N. intertexta MCZ38872

as N intertexta MB20952

N. intertexta PEM R18661

100 N. intertexta MB21183

1.0 N. intertexta PEM R18257

N. ornata NMB R10658

100|I NV. ornata NMB R10907

101 NV omata AMB8635

N. holubi MCZ38793

N. holubi PEM R18647

N. holubi PEM R22814

‘ N. taeniolata PEM R18080

To | NV. taeniolata 2251

N. taeniolata HZ250

N. taeniolata HZ252

N. tessellata AMB5584

N tessellata PEM R16873

N. tessellata PEM R16872

N tessellata PEM R18745

N tessellata NMB R11574

N livida PEM R18747

100 aa

7) N. lvida. MBUR00687

N livida MB21176

N. livida, MB21225

N lida PEM R22822

roy N. lalandii 48037

1.0 N. lalandii, PEM R22815

100 N lalandii. MBUR00483

1.0 N lalandii HZ246

100 N lalandii HB124

1.0 N. lalandii MB20982

100 -— N. broadleyi sp. nov. PEM R24157

Ise N. broadleyi sp. nov. PEM R24005

N. boulengeri_ 1102169

100 70

1.0

A. australis MH0531

A. australis GWO08

100 I capensis CAS 209602

I capensis AMB6001/NMNW

M. suborbitalis PEM R18376

Fig. 2. Maximum likelihood topology for Nucras with bootstrap values (top) and Bayesian posterior probabilities (bottom) at each

node. Bootstrap values <60%, posterior probabilities <O.90, and node support within each species is not shown.

(mitochondrial genes) and two (RAG-1) rate categories,

including the gamma distribution, with uniform priors for

all parameters. For 16S, 38 bases were excluded due to

poor alignment. To ensure the robustness of results, the

MCMC was run twice in parallel for 20 million generations

(four chains in each run), with trees sampled every 1,000

generations. A 10% burn-in was examined (2 million

generations, 2,000 trees) in Tracer v1.6 (http://beast.bio.

ed.ac.uk) to check that the effective sample size (ESS) of

all parameters met a threshold of 200 after burn-in. A 50%

majority rule tree was constructed and nodes with > 0.95

posterior probability were considered supported.

In addition to the Bayesian analysis, a maximum

likelihood (ML) search was run using RAxML HPC 7.2.8

(Stamatakis 2006) on the CIPRES Science Gateway (http://

www.phylo.org/sub_sections/portal/) for the combined

dataset. The datasets were partitioned as in the Bayesian

analysis, with a GTR+I+G model for all markers and 1,000

bootstrap replicates (Stamatakis et al. 2008). This analysis

was run three times to ensure that independent ML searches

produced the same topologies. Nodes with a bootstrap value

Amphib. Reptile Conserv.

of > 70% were considered as supported in this analysis.

Pairwise sequence divergence values (uncorrected

net p-distances) were estimated between species for both

markers using MEGA v7 (Kumar et al. 2016). In addition,

a barcoding approach was used to compare inter- and intra-

specific sequence divergences, using SpeciesIdentifier v1.8

(Meier et al. 2006). Pairwise comparisons were generated

for all Nucras individuals in the phylogeny for each gene,

and frequency distributions of inter- and intra-specific

comparisons were made. The ND4 gene was truncated

433 bp, as some GenBank sequences had only partial

sequences for that gene.

Results

Phylogenetic analyses. The phylogenetic analyses show

that the two individuals from Angola are in the same clade,

and it is sister to a clade containing WN. livida, N. taeniolata,

and N. tessellata (Fig. 2). The new Angolan clade is well-

supported by both Bayesian and likelihood analyses.

Uncorrected net p-distances for each of the genes are

85 November 2019 | Volume 13 | Number 2 | e199

A new Nucras species from Angola

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for Nucras species for a) 16S, b) ND4, and c) RAG-1. Inter-

specific differences shown as black bars, and intra-specific

differences as white bars. The ranges of values relating Nucras

broadleyi sp. nov. with other members of the N. tesse/lata clade

are indicated by brackets.

similar to those found for other species of Nucras (Table

3), and the frequency distribution of pairwise differences

shows that these Angolan individuals fall in the range of

inter-specific divergence values (Fig. 3).

Systematics. Based on the minor morphological

differences and the distinct dorsal coloration differences

observed among material examined, combined with the

abovementioned genetic evidence, the Angolan material

is described below as a new species. No historical names

are available for this clade, thus leaving no outstanding

taxonomic issues (Broadley 1972; Uetz et al. 2017).

Nucras broadleyi sp. nov.

Angolan Sandveld Lizard

urn:lsid:zoobank.org:act: C82E3A75-96FF-4D2A-9B52-3A BF4B58BC2B

Amphib. Reptile Conserv.

(Figures 4-6)

Chersonymy. Nucras tessellata var. taeniolata (Bocage

1895: 30), Nucras tessellata var. taeniolata (Boulenger

1910: 474), Nucras tessellata var. holubi (Boulenger 1917:

210), Nucras intertexta var. holubi (Boulenger 1920: 20),

Nucras tessellata (Monard 1937: 73; Laurent 1964: 56),

Nucras ornata (Broadley 1965: 23), Nucras tessellata

(Broadley 1972: 30; Ceriaco et al. 2016: 56; Burger 2014:

171), Nucras aff. tessellata (Marques et al. 2018: 221;

Branch et al. 2019: 317).

Type material. The type series is comprised of the three

most recently collected specimens, which are housed in

PEM and TM.

Holotype. A subadult male (PEM R24005, AG 018), 10

km west of Lola, edge of Bentiaba River valley, Namibe

Province, Angola (-14.29028, 13.53056, WGS 84, 802 m

asl). Collected by W.R. Branch, P. Vaz Pinto, and J.S. de

Almeida on 2 November 2015.

Paratypes (2). a) A subadult female (PEM R24157,

AG 166), 8.8 km southwest of Farm Mucungo, Namibe

Province, Angola (-14.80167, 12.41917, WGS 84, 385 m

asl). Collected by W.R. Branch, P. Vaz Pinto, and J.S. de

Almeida on 8 November 2015. b) An unsexed adult (TM

40392), “34 km S of Mocamedes to Porto Alexandre,

Angola, 1512Ca” (= 34 km S Namibe to Témbwa),

Namibe Province, Angola (approx. -15.48220, 12.18289).

Collected by W.D. Haacke on 30 March 1971.

Additional referred material: The following additional

material was used to expand the description of variation

within the species: a) an adult male (MD 1967, Laurent

1964), “km 34 de la route de Mocamedes a Sa da Bandeira”

(=34 km from Namibe on Lubango road, -15.03333,

12.41667), collected 24 October 1949, b) MBL 646,

647a, 647b (Bocage 1895: 30) from Maconjo (approx.

-15.01667, 13.20000), c) BM 1970.6.29.10—11 (Boulenger

1910: 474) from Ponang Kuma (= Donguena, approx.

-17.01667, 14.71667), and d) MHNC 91.0524 (Monard

1937) from Capelongo (approx. -14.88333, 15.083333),

collected April 1933.

Etymology. The specific epithet is a patronym tn honor of

Donald G. Broadley for his numerous contributions to the

herpetofauna of Africa. Don (as most of us knew him) was

the first to recognize the Angolan population as a separate

species (Broadley 1972). The name is constructed in the

masculine genitive.

Diagnosis. Assigned to Nucras due to a well-defined collar

(absent in /chnotropis), toes not serrated or fringed (versus

serrated or fringed in Meroles), subdigital lamellae smooth

(versus keeled in Pedioplanis and Heliobolus), subocular

November 2019 | Volume 13 | Number 2 | e199

Branch et al.

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supraciliaries and supraoculars (versus mostly absent in N.

bordering lip, the nostril is pierced between two nasals,

nasal well separated from upper labial, and dorsal scales

small,

boulengeri and N. lalandii), 23-29 lamellae under 4" toe

dorsum with a series of

longitudinal pale stripes (versus dark cross bands present

(versus less than 22 in N. lalandii)

smooth, and juxtaposed.

The new species can be diagnosed from other Nucras

species based on a combination of the following characters:

in N. lalandii and N. scalaris or a series of pale vertebral

spots, sometimes forming irregular transverse bands in N.

intertexta or lack of any dorsal patterns in N. aurantiaca),

series of transversely enlarged plates present under forearm

(versus absent or only feebly enlarged in Nucras lalandii),

four pale stripes on nape with outer stripes forming a

a small series (0-6) of small granules present between

November 2019 | Volume 13 | Number 2 | e199

Amphib. Reptile Conserv.

A new Nucras species from Angola

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SOUIAOIY ade usaseq pIvjowav] SDAINN

DJNUAO

SDAINN

November 2019 | Volume 13 | Number 2 | e199

1998 Ta Wadd

LSc8 lad Wad

CICVET SVO

Lv98 lad Wad

Amphib. Reptile Conserv.

Branch et al.

continuous light stripe with the outer edges of the parietals

(similar to Broadley’s (1972) N. tessellata tessellata var.

“T-” differs from N. livida and N. tessellata where the outer

stripes often do not form a continuous light stripe with the

outer edges of the parietals; differs from N. caeiscaudata

and N. ornata where there are only three longitudinal

stripes present on nape and sometimes the vertebral ones

are absent), well defined occipital scale separating parietals

(versus reduced or absent in northern Namibia N. holubi,

which is referred to as N. intertexta damarana Parker; as

well as absent in NV. caesicaudata), parietal foramen absent

(often present in all other species except N. taeniolata),

and postnasals separated (usually fused in NV. taeniolata).

In the phylogenetic analysis, the uncorrected p-distances

show that this clade differs by >8% for 16S, >14% for

ND4, and >1% for RAGI sequence divergence from other

members of the N. tesse/lata clade.

South Africa, Northern Cape Province

South Africa, Western Cape Province

South Africa, Northern Cape Province

South Africa, Western Cape Province

South Africa, KwaZulu-Natal Province

South Africa, Western Cape Province

Namibia, Kamanjab

Description of Holotype (Fig. 4). Body relatively slender

(SVL approx. 4.5 times the head length, tail truncated),

with hindlimbs larger than forelimbs (femur of hind limb

equal to length of tibia); head narrow and elongated (56%

longer than wide) with narrow pointed but blunt snout, that

is slightly longer than distance from back of eye to rear of

ear opening. Rostrum protruding and visible from below.

Nasals paired and in contact (0.2 mm suture length), not

swollen, nostril directed backwards separating postnasals.

Frontonasal single, wider than long (1.1 x 1.8 mm).

Prefrontals paired and in broad median contact with one

another (0.6 mm suture length), wider than long (1.1 x 1.2

mm). Frontal entire, longer than wide (2.7 x 1.9mm). Two

large rounded supraoculars, both in contact with the frontal,

with anterior supraocular preceded by a single large scale

in contact with prefrontal, frontonasal, and posterior loreal,

with posterior supraocular bordered by a single large

scale in contact with parietal and frontoparietal. Paired

frontoparietal in broad contact (1.3 mm suture length),

nearly as wide as long (1.7 x 1.5 mm). Parietals twice as

long as wide (3.1 x 1.8 mm), fully separate by a large,

pentagonal interparietal (2.5 x 1.2 mm) that is twice as long

as wide, slightly shorter than frontoparietals and nearly

equal to length of frontonasal and prefrontal combined.

Small subtriangular occipital (0.5 x 0.7 mm). Two loreals,

second much larger than first. Six supraciliaries on each

side, 1‘ is the longest. A single minute granule scale

between supraocular and supracilliares on right side, none

on left side. Four supralabials anterior to subocular and

three supralabials posterior to subocular, on both sides.

Subocular slightly elevated medial and bordering the

lip, its lower border being shorter than the upper. Three

temporal scales, first longer than others, smooth. Tympanic

shield as wide as long, border of ear opening. No ear lobes.

Lower eyelid with transparent brille formed by five larger

scales, surrounded by numerous smaller scales. Lower

eyelid separated from subocular and enlarged temporal

scales by a series of 10 smaller scales. Small scale above

3 supralabial separating the posterior loreal and subocular.

accession number

Museum

Field accession

Australolacerta

Table 2 (continued). Samples used in genetic analysis. Museum abbreviations: CAS — California Academy of Science, PEM — Port Elizabeth Museum, NMB — National Museum Bloemfontein.

Australolacerta

)

Lae!

Amphib. Reptile Conserv. 89 November 2019 | Volume 13 | Number 2 | e199

A new Nucras species from Angola

Table 3. Pairwise uncorrected net p-distances for species of Nucras: a) 16S, b) ND4, c) RAG-1. Comparisons not made due to

missing data indicated by na.

a) 1 2 3

1 broadleyi sp. nov 0.048

2 lalandii 0.083 0.047

3 livida 0.097 0.058 0.019

4 taeniolata 0.098 0.056 0.084

5 tessellata 0.075 0.043 0.026

6 boulengeri 0.105 0.101 0.117

7 holubi 0.074 0.044 0.066

8 intertexta 0.084 0.068 0.090

9 ornata 0.083 0.067 0.091

1 broadleyi sp. nov na

2 lalandii 0.139 0.124

3 livida 0.194 0.115 0.045

4 taeniolata 0.208 0.147 0.121

5 tessellata 0.198 0.138 0.112

6 boulengeri 0.219 0.188 0.233

7 holubi 0.153 0.092 0.137

8 intertexta 0.208 0.141 0.175

1 broadleyi sp. nov 0.001

2 lalandii 0.012 0.002

3 livida 0.014 0.009 0.004

4 taeniolata 0.013 0.007 0.009

5 tessellata 0.012 0.006 0.008

6 boulengeri 0.066 0.059 0.064

7 holubi 0.020 0.015 0.017

8 intertexta 0.017 0.012 0.013

9 ornata 0.021 0.016 0.016

4 5 6 7 8 9

0.000

0.073 0.027

0.116 0.103 na

0.063 0.053 0.079 0.048

0.085 0.078 0.105 0.043 0.008

0.082 0.076 0.108 0.050 0.043 0.000

0.003

0.006 0.018

0.277 0.267 na

0.156 0.146 0.199 0.115

0.196 0.193 0.239 0.118 0.017

0.004

0.007 0.008

0.064 0.058 na

0.016 0.016 0.068 0.004

0.012 0.013 0.064 0.015 0.001

0.016 0.017 0.064 0.019 0.004 0.001

Enlarged scale bordering 1“ post subocular, supralabial,

and the subocular. Six infralabials on both sides, with

3" being longest; four enlarged pairs of chin shields,

last largest and first three in broad contact. Twenty-four

gular scales in a straight line between symphysis of chin

shields and median collar plate, equal in size except last

4—5 larger. Collar free, comprising seven enlarged plates

(median subtriangular) and extending slightly onto side of

neck as a crease, bordered by 2—3 smaller scales. Dorsal

scales small, juxtaposed, granular, smooth, larger on sides

toward ventrals. Midbody scales 42. Ventral plates eight

longitudinal and 28 transverse rows (from collar to groin),

plates of the innermost rows longer than broad, with outer

row notably smaller than other rows, transverse row of

ventrals across chest just behind collar longer than broad:

preanal scales irregular, median ones larger. Scales on

upper surface of forearm large, smooth or slightly keeled.

Scales on lower surface of forearm with eight enlarged

plates, at least twice the width of scales on upper forearm.

Amphib. Reptile Conserv.

Scales on upper surface of tibia rhombic, subimbricate,

smooth, and much larger than dorsal scales. Tibia below

with a series of large plates. Subdigital lamellae under

fourth toe 23R/25L. Femoral pores 13R/15L. Dorsal scales

on tail oblique, strongly keeled diagonally, and truncate

behind, ventral scales on tail obtusely keeled.

Coloration. Dorsum with eight pale cream to white

dorsolateral longitudinal stripes, separated by dark brown

to black stripes. These stripes are more boldly patterned

anteriorly, fading posteriorly. No light vertebral stripe.

The two pale paravertebral stripes are separated by a very

narrow strip of darker scales that starts on the interparietal

through the occipital scale and fades posteriorly onto body

and tail. The dorsolateral stripe extending along outer

borders of parietals continues onto the tail. It is followed

by the upper lateral stripe extending from posterior of the

eye onto the head through the mid-temporal with a brief

break above the ear opening, and continues onto the tail.

The lower lateral stripe starts at the subocular, through the

November 2019 | Volume 13 | Number 2 | e199

Branch et al.

nak acl; Var red: bee M7 pet

Fig. 4. Nucras broadleyi sp. nov. A—

holotype, adult male, PEM R24005 (AG 18) in life; B— general habitat photo of type locality,

tre

-y" =

tetet tht beter fa!

10 km west of Lola, edge of Bentiaba River valley, Namibe Province, Angola; C — lateral close-up of head of holotype; D — dorsal

close-up of head of holotype; E — ventral close-up of head of holotype (Photos: Bill Branch).

ear opening, broken briefly above the arm, after which

it continues all the way onto the tail. Ventrum white and

lower limbs oblique white. Fore limbs upper surface black

with scattered pale blotches. Hind limbs light brown with

pale blotches. Upper surface of tail red-brown, similar to

hind limbs. Scales bordering the orbit are black edged.

Variation (Figs. 5-6). Meristic and escalation data are

summarized in Table 1. The largest specimen examined

is (BM 1907.6.29.10) 74 +144 mm (tail regenerated).

Regarding coloration, there seem to be three main variations

among material examined: 1) 8—9 longitudinal stripes as

in holotype (in PEM R24005, MBL 647a, 647b, MHNC

91.0524—5), 2) 4-5 pale longitudinal stripes broken up

posteriorly with flanks spotted (in BM 1970.6.29.10-11,

TM 40392, MD 1967), and 3) broken paravertebral stripes,

continuous dorsolateral line and barred flanks (in PEM

R24157), similar to N. intertexta.

Distribution. Found only in semi-arid south-western

Angola, throughout much of Namibe Province and

extending onto the escarpment of southern Huila and

Cunene Provinces (Fig. 1). Known localities include:

Maconjo (Bocage 1895: 30), Ponang Kuma (=Donguena)

Amphib. Reptile Conserv.

(Boulenger 1910: 472), 34 km from Namibe on Lubango

road (Laurent 1964: 56), 34 km south of Tombwa (TM

40397), 8.8 km southwest of Farm Mucungo (this study),

10 km west of Lola (this study), and Capelongo (Monard

1937: 73). The locality of Caconda (Bocage 1895) extends

the species distribution further north into Huila Province,

but the specimens could not be critically evaluated by

Broadley (1972) and are now presumably lost.

Habitat. The species appears to be associated with

mopane woodlands, dry savannas, and semi-desert

shrublands (Barbosa 1970). The new material was found

in sandy plains with scattered low granite outcrops, with

varying degrees of short grass cover and scattered bushes.

Vegetation included Colophospermum mopane, Ficus sp.,

Senegalia (=Acacia) mellifera, Commiphora sp., Boscia

foetida, and Salvadora persica. The confirmed historical

records were also obtained within the dry woodland zone,

even though the possible occurrence of the species in

Caconda would place the species above 1,500 m asl and

well into the mesic conditions of Brachystegia habitats

(Barbosa 1970).

Conservation. Population estimates for the species

November 2019 | Volume 13 | Number 2 | e199

A new Nucras species from Angola

- th TWh olt hei ‘even

Fe a

iui

Fig. 5. Nucras broadleyi sp. nov. A — paratype, adult female, PEM R24157 (AG 166) dorsal view; B — ventral view; C — dorsal

close-up of head of paratype; D — ventral close-up of head of paratype; E — lateral close-up of head of paratype; F — general habitat

photo of type locality, 8.8 km southwest of Farm Mucongo, Namibe Province, Angola (Photos: Bill Branch).

are unknown, and only few scattered specimens (~12)

are known, of which four specimens were destroyed in

the Museu Bocage Lisboa fire and one of the Monard

Specimens is unaccounted for. However, Sandveld Lizards

are secretive and less conspicuous than many other

lacertids, so additional surveys are required to determine

the full range of the species and to identify potential habitat

threats in order to accurately assess its conservation status.

Discussion

Broadley (1972) was the first to suggest the Angolan pop-

ulation of Nucras tessellata to be different from other de-

scribed species, but took no taxonomic action. Here, we

present evidence to support his assumptions and formally

describe the Angolan population as a new species. Thus,

Angola now has two endemic species of Nucras and the

genus now comprises 12 recognized species. As our phy-

logeny is built on the work of Edwards et al. (2013) we

retrieved the same general topology, except for the inclu-

sion of the new species. Although different samples and

genetic markers were used, Bauer et al. (2019) retrieved

the same species relationships except for the inclusion

of their newly described species, N. aurantiaca. Thus,

we can conclude that the current species relationships are

Amphib. Reptile Conserv.

well resolved. Due to the secretive nature of members

of this genus, disjunct distribution, and previously rec-

ognized varieties (see Broadley 1972), it is possible that

there are other undiscovered species, particularly in areas

that remain poorly surveyed.

The species appears restricted to the arid biomes of

southwestern Angola at relatively low to moderate alti-

tudes, while the records from Caconda remain problematic

and may have been misidentified or incorrectly labelled.

In recent years, the number of endemic species described

from the arid south-western Angola has increased, e.g.,

Kolekanus plumicaudus (Haacke 2008), Pedioplanis

huntleyi and P. haackei (Conradie et al. 2012), Cordy-

lus namakuiyus (Stanley et al. 2016), Cordylus phono-

lithos (Marques et al. 2019b), Poyntophrynus pachnodes

(Ceriaco et al. 2018), and now Nucras broadleyi sp.

nov. This region also harbors numerous other endemic

species, such as Afrogecko ansorgii, Pachydactylus an-

golensis, Poyntonophrynus grandisonae, Pedioplanis

benguellensis, Rhoptropus taeniostictus, Typhlacontias

rudebecki, and T. punctatissimus bogerti (Ceriaco et al.

2016, 2018; Marques et al. 2018; Branch et al. 2019).

The growing body of information suggests there could

be a unique and diverse endemic Angolan-Namib reptile

fauna (Ceriaco et al. 2016; Marques et al. 2018; Branch

November 2019 | Volume 13 | Number 2 | e199

Branch et al.

Fig. 6. Variation of Nucras broadleyi nov. sp. dorsal color pattern. A—TM 40392 from “34 km S of Moc¢amedes to Porto Alexandre;”

B — BM 1970.6.29.10 from Ponang Kuma (=Donquena); C - MHNC 91.0524 from Capelongo; D— MD 1967 from “km 34 de la

route de Mocamedes a Sa da Bandeira” (Photos: A,B — Bill Branch, C, D — Luis Ceriaco).

et al. 2019), with additional discoveries yet to be made.

Acknowledgements.—We thank José Luis Alexandre

and Fernanda Lages for organizing the export permits

for the vouchers, done within the framework of a

SASSCAL Project sponsored by the Federal Ministry

of Education and Research (BMBF) - ISCED permit

issued 12 November 2015. We thank Lemmy Mashinini

(Ditsong Museum, Pretoria, South Africa) and Patrick

Campbell (Natural History Museum, London, United

Kingdom) for allowing WRB to inspect material in their

care. Shiela Broadley kindly provided access to the late

Don Broadley’s data, noting that Don was one of the last

researchers that managed to study the Bocage material

before a fire destroyed the collection. Aaron Bauer and

Luis Ceriaco are thanked for providing additional data

and comparative photographs of Angolan material, and

their excellent reviews which improved the quality of

this paper. Jodo Simdes de Almeida is thanked for his

field assistance. This work was supported in part by the

National Research Foundation of South Africa and the

South African National Biodiversity Institute.

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Bill Branch (William R. Branch) was born in London, United Kingdom. Bill was

employed as Curator of Herpetology at the Port Elizabeth Museum for over 30 years

(1979-2011), and upon his retirement he was appointed Curator Emeritus Herpetology

until his death in October 2018. Bill’s herpetological studies concentrated mainly

on the systematics, phylogenetic relationships, and conservation of African reptiles,

but he has been involved in numerous other studies on the reproduction and diet of

African snakes. He has published over 300 scientific articles, as well as numerous

popular articles and books. The latter include: South African Red Data Book of

Reptiles and Amphibians (1988), Dangerous Snakes of Africa (1995, with Steve

Spawls), Field Guide to the Reptiles of Southern Africa (1998), Tortoises, Terrapins,

and Turtles of Africa (2008), and Atlas and Red Data Book of the Reptiles of South Africa, Lesotho, and Swaziland (multi-authored,

2014), as well as smaller photographic guides. In 2004, Bill was the 4" recipient of the “Exceptional Contribution to Herpetology”

award of the Herpetological Association of Africa. Bill has undertaken field work in over 16 African countries, and described nearly

50 species, including geckos, lacertids, chameleons, cordylids, tortoises, adders, and frogs.

Amphib. Reptile Conserv.

Werner Conradie holds a Masters in Environmental Science (M. Env. Sc.) and has 12 years of

experience with the southern African herpetofauna, with his main research interests focusing on the

taxonomy, conservation, and ecology of amphibians and reptiles. Werner has published numerous

principal and collaborative scientific papers, and has served on a number of conservation and

scientific panels, including the Southern African Reptile and Amphibian Relisting Committees.

He has undertaken research expeditions to many African countries including Angola, Botswana,

Lesotho, Malawi, Mozambique, Namibia, South Africa, Zambia, and Zimbabwe. Werner is currently

the Curator of Herpetology at the Port Elizabeth Museum (Bayworld), South Africa.

Pedro Vaz Pinto is Angolan and was born in Luanda, Angola, in 1967. Pedro graduated in Forest

Engineering at the Technical University of Lisbon, and obtained a doctoral degree in Biology from

the University of Porto, Portugal. Over the past 20 years, he has worked in biodiversity conservation

in Angola addressing rare or endangered species, and protected area management. Pedro is a director

for the local NGO Kissama Foundation, and a researcher for CIBIO-InBio. His studies on Angolan

vertebrates have focused mostly on genetics, biogeography, and conservation in antelopes, birds,

reptiles, and amphibians. Pedro travels the country extensively and has received three international

environmental awards for his biodiversity conservation work in Angola.

Krystal Tolley is a Principal Researcher at the South African Biodiversity Institute in South Africa.

Krystal studies patterns of biodiversity and adaptation of African reptiles by combining phylogenetics,

phylogeography, performance-based data, species distribution models, and morphology.

November 2019 | Volume 13 | Number 2 | e199

Official journal website:

amphibian-reptile-conservation.org

Amphibian & Reptile Conservation

13(2) [Special Section]: 96-130 (e203).

The herpetofauna of Bicuar National Park and surroundings,

southwestern Angola: a preliminary checklist

12.3*Ninda L. Baptista, '¢Telmo Antonio, and °°tWilliam R. Branch

‘Instituto Superior de Ciéncias da Educagao da Huila (ISCED-Huila), Rua Sarmento Rodrigues, Lubango, ANGOLA *CIBIO/InBio — Centro de

Investigagdo em Biodiversidade e Recursos Genéticos, Universidade do Porto, Campus de Vairdo, 4485-661 Vairdo, PORTUGAL *Faculdade

de Ciéncias, Universidade do Porto, 4169-007 Porto, PORTUGAL ‘Faculty of Natural Resources and Spatial Sciences, Namibia University of

Science and Technology, Private Bag 13388, Windhoek, NAMIBIA *Port Elizabeth Museum (Bayworld), P.O. Box 13147, Humewood 6013, SOUTH

AFRICA ‘Research Associate, Department of Zoology, P.O. Box 77000, Nelson Mandela Metropolitan University, Port Elizabeth 6031, SOUTH

AFRICA * Deceased

Abstract.—Bicuar National Park (BNP) is a protected area in southwestern Angola where biodiversity has

been poorly studied. BNP is located on the Angolan plateau on Kalahari sands, in a transition zone between

the Angolan Miombo Woodland and the Zambezian Baikiaea Woodland ecoregions. Herpetological surveys

were conducted in BNP and surrounding areas, through visual encounter surveys, trapping, and opportunistic

collecting of specimens from 2015 to 2018. The regional herpetofauna is described here based on these

surveys, literature records, and additional unpublished records. In total, 16 amphibian, 15 lizard, 18 snake,

two testudine, and one crocodilian species were observed from the recent surveys, and in combination with

historical records the species counts are 21, 36, 32, four, and one species for these herpetofauna groups,

respectively. Important observations include the first record of Xenocalamus bicolor bicolor (Gunther, 1868),

the second records of Sclerophrys poweri (Hewitt, 1935) and of Amblyodipsas ventrimaculata (Roux, 1907),

and the fourth record of Monopeltis infuscata (Broadley, 1997) for Angola. Additionally, the type locality of

Hyperolius benguellensis (Bocage, 1893) is discussed. A part of the material could not be confidently identified

to species level, reflecting the taxonomic uncertainty associated with the Angolan herpetofauna. Fossorial

herpetofauna was well represented, reflecting adaptation to sandy soils, the dominant substrate in the area.

The likely presence of endemic and poorly known species in BNP reinforces the importance of the park for the

conservation of Angolan biodiversity. Further surveys are necessary for a more comprehensive understanding

of the park’s fauna and biogeographic affinities, and to improve conservation planning.

Keywords. Amphibians, reptiles, fossorial, biodiversity surveys, protected areas, Kalahari sands, Huila Province

Resumo.—O Parque Nacional do Bicuar (BNP) é uma area protegida no sudoeste de Angola cuja biodiversidade

se encontra pouco estudada. Localiza-se no planalto de Angola em areias do Calaari, numa zona de transigao

entre as ecorregides de Mata de Miombo Angolana e Mata de Baikiaea Zambeziana. Neste trabalho foram

realizados levantamentos de herpetofauna no BNP e arredores, através de levantamentos de encontro visual,

armadilhagem e recolha oportunistica de espécimes entre 2015 e 2018. Aqui é€ apresentada uma descricgao

da herpetofauna da regiao baseada nestes levantamentos, em registos bibliograficos, e outros registos nao

publicados. Os dados recentes resultaram num total de 16 espécies de anfibios, 15 espécies de lagartos, 18

especies de cobras, duas espécies de quelonios, e uma espécie de crocodilo. A combinagao destes dados

com registos historicos resulta num total de 21, 36, 32, quatro, e uma espécies destes grupos herpetoldgicos,

respectivamente. Entre os resultados mais importantes estao o primeiro registo de Xenocalamus bicolor bicolor

Gunther, 1868, o segundo registo de Sclerophrys poweri (Hewitt, 1935) e de Amblyodipsas ventrimaculata

(Roux, 1907), e o quarto registo de Monopeltis infuscata Broadley, 1997 para Angola. A localidade-tipo de

Hyperolius benguellensis (Bocage, 1893) € tambem discutida. Uma parte do material nao pdde ser identificado

com certeza ao nivel da espécie, uma consequéncia da incerteza taxonomica associada a herpetologia

angolana. A herpetofauna fossorial esta bem representada, reflectindo uma adaptacgao a solos arenosos, o

substrato dominante na area. A presenga provavel de espécies endemicas e pouco conhecidas no BNP reforca

a importancia do parque para a conservagcao da biodiversidade de Angola. Mais levantamentos contribuirao

para um melhor conhecimento da fauna do parque e das suas afinidades biogeograficas e para um melhor

planeamento de estrategias de conservagao.

Palavras-chave. Anfibios, répteis, fossorial, levantamentos de biodiversidade, areas protegidas, areias do Calaari,

provincia da Huila

Correspondence. * nindabaptista@gmail.com

Amphib. Reptile Conserv. 96 December 2019 | Volume 13 | Number 2 | e203

Baptista et al.

Citation: Baptista NL, Antonio T, Branch WR. 2019. The herpetofauna of Bicuar National Park and surroundings, southwestern Angola: a preliminary

checklist. Amphibian & Reptile Conservation 13(2) [Special Section]: 96—130 (e203).

4.0 International (CC BY 4.0): https://creativecommons.org/licenses/by/4.0/], which permits unrestricted use, distribution, and reproduction in any

medium, provided the original author and source are credited. The official and authorized publication credit sources, which will be duly enforced, are

as follows: official journal title Amphibian & Reptile Conservation; official journal website: amphibian-reptile-conservation.org.

Received: 18 August 2018; Accepted: 11 November 2019; Published: 3 December 2019

Introduction

Angola’s biodiversity is poorly understood, especially

when compared to other countries in southern Africa

(Huntley et al. 2019b). A recent synthesis of the country’s

biodiversity (Huntley et al. 2019b) provides updated

checklists for several taxonomic groups, including

amphibians (Baptista et al. 2019) and reptiles (Branch

et al. 2019c), as does the historical atlas by Marques et

al. (2018), but all these lists highlight the understudied

status of the herpetofauna in Angola. Southwestern

Angola is one of the better-studied regions for vertebrates

in the country (Crawford-Cabral and Mesquitela 1989),

and this is reflected in herpetofauna, from which several

type descriptions originated. Despite many historical

herpetological surveys in the area (Bocage 1895; Schmidt

1933, 1936; Monard 1937a,b; Mertens 1938; Bogert

1940; Laurent 1964; Gans 1976; Poynton and Haacke

1993), contemporary studies continue to yield important

discoveries of new species (Haacke 2008; Conradie

et al. 2012, 2013; Stanley et al. 2016; Ceriaco et al.

2018b; Branch et al. 2019b), new species records for the

country (Huntley 2009; Ceriaco et al. 2016; Branch et al.,

unpub. data), and rediscoveries of species not observed

for several decades (Baptista et al. 2018; Branch et al.

2019a; Vaz Pinto et al. 2019).

The herpetofauna remains undersampled — or

unsurveyed in vast areas of southwestern Angola,

including Bicuar National Park (BNP). Wulf Haacke

conducted two significant herpetological surveys in

this region in 1971 and 1974, collecting over 2,000

specimens (Branch et al. 2019c). Although BNP was not

specifically sampled by Haacke, similar habitats near the

park were included in the surveys. Poynton and Haacke

(1993) described the amphibian collection resulting

from these surveys, but information on the new and rare

reptiles deposited in the Ditsong National Museum of

Natural History (former Transvaal Museum) was never

formally published (Branch et al. 2019c). The Angolan

civil war prevented research in the country for 35 years

(1974-2009) between Haacke's expeditions and the first

post-war biodiversity surveys in southwestern Angola

(Huntley 2009). The first herpetological surveys inside

BNP were part of this effort, during a brief visit by Alan

Channing. Only two adult Hyperolius benguellensis and

several species of tadpoles were collected (Channing et

al. 2013; A. Channing, pers. comm.), with the material

deposited in Berlin Zoological Museum, and other

Amphib. Reptile Conserv.

of,

observations remained unpublished. More recent surveys

include two short visits to BNP in 2017 and 2018 by

Butler et al. (2019).

To document the herpetofauna of BNP, we conduct-

ed surveys in and around the park from 2015 to 2018

as part of the Southern African Science Service Centre

for Climate Change and Adaptive Land Management

(SASSCAL) project, which established an observa-

tory in BNP. This is part of a network of five other

Angolan observatories, and a total of 47 observatories

throughout Southern Africa (Jurgens et al. 2018). Re-

sults of herpetological surveys in Tundavala observa-

tory, southwestern Angola, were published recently

(Baptista et al. 2018). This paper draws upon records

and photographs of BNP herpetofauna partially avail-

able since 2017 through SASSCAL’s on-line platform

(SASSCAL ObservationNet 2018), and is their first

formal publication. It presents the results of the 2015-—

2018 SASSCAL surveys, and it includes collated data

from recent and historical collections and observations

made by others in and around BNP to provide a more

comprehensive account.

Materials and Methods

Study Area

Bicuar National Park (BNP) in Huila Province,

southwestern Angola, was declared as a Partial Game

Reserve in 1957 to protect populations of big game

before being upgraded to National Park status in 1964

(Teixeira 1968; Huntley et al. 2019a). The park was

originally 790,000 ha in size (Huntley et al. 2019a), with

boundaries described in Diploma Legislativo n° 3527

of 26 December 1964 (Teixeira 1968; Simdes 1971).

In 1972, a governmental decree in Portaria 384/72

deproclaimed areas of northern BNP for the expansion

of the “Capelongo colonial settlement” (“Colonato de

Capelongo”) [L. Verissimo, pers. comm.], defined the

current boundaries of BNP, and decreased the park’s

area to 675,000 ha (Mendelsohn and Mendelsohn 2018;

Huntley et al. 2019a).

BNP lies at ca. 1,200-1,400 m above sea level (asl),

between the Cunene and Caculuvar rivers on wind-

blown Kalahari sands (Mendelsohn and Mendelsohn

2018). This is the most extensive contiguous body of

sand in the world (Leistner 1967) and extends from the

southern African plateau to the Congo basin. The park

December 2019 | Volume 13 | Number 2 | e203

Herpetofauna of Bicuar National Park, Angola

Teles tS a

Fig, 1. Habi

tat types in the study area. (A) Angolan Miombo woodland on Kalahari sands, BNP; (B) Burkea/Baikiaea woodlands,

Carmira Farm; (C) dry grassy wetland (“mulola”) flanked by miombo woodlands (“tunda”), BNP; (D) temporary pan in a “mulola,”

BNP.

is part of the “Lower Cunene” mesological unit (Diniz

2006; Huntley 2019), and is located in a transition

zone from moist to dry savannas. It comprises both the

Angolan Miombo Woodland and Zambezian Baikiaea

Woodland ecoregions, as defined by Burgess et al.

(2004), which are equivalent to the Brachystegia and

South-west Arid biomes, respectively, as characterized

by Huntley (1974). The northern extent is dominated by

miombo woodlands, consisting mostly of Brachystegia

spiciformis and Julbernardia paniculata, while the south

is covered predominantly by savannas dominated by

Burkea africana (Teixeira 1968) [Fig. 1], and Angolan

Mopane woodlands are present to the south of the park

(Huntley 2019). In addition to woodlands, thickets, and

scrublands of varied composition, open drainage lines

hosting grasslands with geoxylic suffrutex shrublands

are common throughout; see Teixeira (1968), Barbosa

(1970), and Chisingui et al. (2018) for further details.

Elliptical in shape, the park is ca. 80 km in diameter

from north to south and ca. 110 km from east to west

(Fig. 2). The climate is seasonal, with precipitation

falling mainly from October to April, and nocturnal frost

occuring frequently in the dry season, especially in June

and July. Meteorological data for BNP observatory are

available from 2015 onwards (SASSCAL WeatherNet

Amphib. Reptile Conserv.

2019). The average annual rainfall varies from ca.

900-950 mm in the northern border and 650-700 mm

in the southern border, and average annual temperatures

vary between ca. 19-20 °C in the north and 22—23 °C

in the south, with values for north and south taken from

Quipungo and Mulondo, respectively, in Mendelsohn

and Mendelsohn (2018).

Topographically BNP is generally flat, interrupted by

relatively parallel drainage lines mostly flowing west to

east (Fig. 2). The park is divided through the center by

a larger depression, called Bicuar, which flows in the

north-south direction and forms part of the Cunene River

catchment (Teixeira 1968). Natural grasslands with

geoxylic suffrutex shrublands occur in drainage lines that

are seasonally filled with water (locally called “mulolas”’)

[Fig. 1D] where cold air accumulates at night during the

dry season. Many of the existing permanent water bodies

in the park are artificial excavations made to attract game

for observation purposes (Simdes 1971). The park’s

elevated regions (locally called “tundas’’) are covered by

woodlands, and are only 30-50 m higher in altitude than

the valleys with grasslands. BNP’s soils are mostly sandy

(Missao de Pedologia de Angola 1959; Teixeira 1968),

and arenosols as defined by Jones et al. (2013), with

rare rocky outcrops. The landscape surrounding BNP

December 2019 | Volume 13 | Number 2 | e203

Baptista et al.

Handa Farm

Quipungo

Capelongo

Chibemba

Carmira Farm:

14°0'E 15°0'E

i is‘o’s

H— 16°0'S

0 2620 ma.s.l.

— Bicuar National Park boundaries

O Historical records (before 1975)

_ Recent records from Butler et al. (2019)

A Recent records from this study (after 2008)

16°0'E

Fig. 2. Left: Bicuar National Park (BNP) and surveyed sites from historical and recent records. Right: Angola with provincial

boundaries and location of BNP. Source of satellite image: Maxar (https://www.digitalglobe.com/).

is similar (Barbosa 1970; Mendelsohn and Mendelsohn

2018) such that records from localities surrounding the

park are also reported in this work.

Survey Methods and Data Sources

For this study, surveys of herpetofauna were performed

in BNP from October 2015 to April 2018. These included

opportunistic collections during occasional visits to the

park, and two series of focused surveys, from 2-10

December 2016 (during the rains) and 3-7 November

2017 (at the onset of the rains). For both surveys, diurnal

and nocturnal Visual Encounter Surveys (VES) were

performed, as well as dipnetting in water bodies by

scooping the bottom, using nets of varied shape and mesh

size. Trap arrays consisting of drift fences with pitfall and

funnel traps were set during the 2016 survey at three sites

within the following habitats: miombo woodlands not

burnt for more than one year (site T1, see Appendix | for

coordinates and duration), miombo woodlands not burnt

for more than five years (T2, Appendix 1), and grassland

along a drainage line (T3, Appendix 1). Each trap array

consisted of one plastic drift fence (15 m long and 50 cm

high) with two pitfall traps, one at each end of the drift

fence, and six funnel traps placed on adjacent sides of

the fence. Collecting sites in BNP and the surroundings

included the Main Camp and permanent water bodies

(waterhole near the Main Camp, Lagoa da Matemba,

Lagoa do Djimbi, and Lagoa Nougalafa, among others).

These sites are mapped in Fig. 2, and geographic

Amphib. Reptile Conserv.

coordinates are provided in Appendix 1. Surveyed habitat

types included miombo and Burkea/Baikiaea woodlands,

ponds, and excavations in drainage lines where water is

permanently provided by water pumps. Large portions

of the surveyed areas in the north were burnt two to four

weeks prior to being surveyed.

Additional observations include records from

Channing’s 2009 visit (A. Channing, pers. comm.), as

well as opportunistic records from other researchers

working in the region (Manfred Finckh and Francisco

Maiato, who provided photographs), and from two farms

located near the park. Carmira Farm, located ca. 40 km

northeast of Cahama and ca. 46 km south of BNP (site CF),

has staff with a personal interest in collecting reptiles, as

they have created a collection (Fig. 3) comprising snakes,

amphisbaenians, and writhing skinks. Handa Farm, 30

km northwest of Quipungo and 48 km northwest of BNP

(site HF), maintains a collection of photographic records

of its fauna, including reptiles. All recent records in these

sources were considered for this study, and all except

Channing’s 2009 data (A. Channing, pers. comm.) were

verified by the authors. Butler et al. (2019) addressed

BNP herpetofauna, and these results have also been

incorporated into this work.

Collected specimens were photographed and

euthanized by either submersion (in the case of frogs) or

injection into the intracoelomic cavity (for reptiles) of a

solution of tricaine methanesulfonate (MS222) [Conroy

et al. 2009]. Tissue (liver or muscle) was preserved in

99.5% ethanol for genetic analysis. Specimens were fixed

December 2019 | Volume 13 | Number 2 | e203

Herpetofauna of Bicuar National Park, Angola

Fig. 3. Carmira Farm private collection.

with formalin, then transferred to water (to remove the

formalin), before finally being transferred to 70% ethanol

for long-term storage. All photographs were taken by the

first author (N. Baptista), except when noted. Specimens

are held in the herpetological collection of Instituto

Superior de Ciéncias da Educacao da Huila (ISCED-

Huila), Lubango, Angola. Additional specimens that

were not collected for this study are deposited in Carmira

Farm’s private collection (Fig. 3).

Field guides (Branch 1998; Schigtz 1999; Channing

2001; Marais 2004; du Preez and Carruthers 2009;

Channing et al. 2012; Channing and Rodel 2019) were

used for species identification, and additional taxon-

specific literature was consulted when necessary.

Nomenclature followed online databases: Amphibian

Species of the World (Frost 2019) for amphibians, and

The Reptile Database (Uetz et al. 2019) for reptiles, and

was updated when appropriate. Only materials from the

ISCED-Huila collection and Carmira and Handa Farms

were examined for this study. Taxonomic identification

of historical records (published in the literature, and

unpublished records from Haacke’s surveys) and acoustic

survey records may require verification.

Localities for historical data compiled for the BNP

region (yellow dots in Fig. 2) were selected based

on distances to the park (1.e., those within a 100 km

radius of the park’s boundaries) and physiographic

similarity (Huntley 2019). The only exception to this

is Humbe, which is located 110 km south of the park,

but was also included in this account since it is an

important historical collection site and is the type

locality of several reptile species. Selected historical

Amphib. Reptile Conserv.

localities were: Cahama and surroundings, Capelongo

(= Kapelongo, = Folgares), Catequero and surroundings,

Chibemba and surroundings, Dongue and surroundings,

Gambos (= Chibemba, see discussion in Branch et

al. [2019a]), Humbe, Humbi (= Humbe), Humbia (=

Humbe), Kandingu (= Kului River), Kangela (= Kului

River), Kului River, Mulondo, Mupa, Osi (= Osse),

Osse, Quipungo, and Viriambundo. Records from these

localities were compiled from published literature such

as Bocage (1895), Monard (1937a,b), Schmidt (1933),

Bogert (1940), Laurent (1964), Gans (1976), Poynton

and Haacke (1993), and from unpublished information

in the Ditsong (=Transvaal) National Museum of Natural

History's database (Haacke, TM).

Collected data were grouped into three classes (Fig.

2): 1) historical records collected before 1975, which

include records published in historical literature and

unpublished records from TM; 11) recent records from

Butler et al. (2019); and 11) recent records from this

study, which refer to either data collected during field

surveys from 2009 onwards performed within the

scope of the SASSCAL project, A. Channing’s personal

records, opportunistic photographic records collected by

other researchers in the region, and records from Carmira

Farm and Handa Farm.

Results

The combined records (published and unpublished, recent

and historical) of herpetofauna biodiversity from the

BNP region comprises a total of 94 taxa. This includes 21

amphibian taxa (Table 1) and 73 reptilian taxa (36 lizards,

December 2019 | Volume 13 | Number 2 | e203

Baptista et al.

Table 1. Amphibians recorded from inside and the surroundings of Bicuar National Park, Angola, based on historical and recent

records. Type of record: C = advertisement call; L = literature; O = observation; P = photograph; RR = new record for the region; V

= voucher. Period of record: A = after 2008; B = before 1975. Taxonomy has been updated over the years, therefore original species

citations may occur under different names.

10.

11.

12.

13.

14.

es:

16.

17.

18.

Species

Arthroleptidae

Leptopelis bocagii (Gunther, 1865)

Bufonidae

Mertensophryne aff. mocquardi (Angel, 1924)

Sclerophrys poweri (Hewitt, 1935)

Sclerophrys pusilla (Mertens, 1937)

Sclerophrys regularis (Reuss, 1833)

Hemisotidae

Hemisus cf. guineensis Cope, 1865

Hyperoliidae

Hyperolius angolensis Steindachner, 1867

complex

Hyperolius benguellensis (Bocage, 1893) complex

Kassina senegalensis (Duméril and Bibron 1841)

Microhylidae

Phrynomantis bifasciatus (Smith, 1847)

Phrynobatrachidae

Phrynobatrachus mababiensis FitzSimons, 1932

Phrynobatrachus natalensis (Smith, 1849)

Pipidae

Xenopus petersii Bocage, 1895

Ptychadenidae

Hildebrandtia cf. ornata (Peters, 1878)

Ptychadena ansorgii (Boulenger, 1905)

Ptychadena oxyrhynchus (Smith, 1849)

Ptychadena porosissima (Steindachner, 1867)

Pyxicephalidae

Pyxicephalus adspersus Tschudi, 1838

Amphib. Reptile Conserv.

Type of

record

RR, V

Lev.

Le @

101

Records in the region of BNP _ Inside

Locality (Reference) BNP?

BNP (this study) Y

Mulondo (Monard 1937a)

BNP (this study)

Capelongo (Butler et al. 2019);

BNP (this study)

Humbe, Mulondo, Mupa, Osi (=

Osse) (Monard 1937a)

BNP (this study) Y

Capelongo, Osi (= Osse)

(Monard 1937a); BNP (Butler et Y

al. 2019; this study)

BNP (Channing et al. 2013);

BNP (Butler et al. 2019; this Y

study)

Mulondo (Monard 1937a); BNP Y

(this study)

Mulondo (Monard 1937a); BNP Y

(this study)

BNP (Butler et al. 2019; this Y

study)

Kangela (Monard 1937a); BNP Y

(this study)

Kandingu, Osi (= Osse)

(Monard 1937a); BNP (Butler et Y

al. 2019; this study)

Mulondo, Osi (= Osse) (Monard

1937a); Dongue (Poynton

and Haacke 1993); between

Chibemba and Cahama (this

study)

Kandingu (Monard 1937a)

Osi (= Osse) (Monard 1937a);

BNP (Butler et al. 2019; this Y

study)

BNP (this study) Y

Humbe (Bocage 1895); Carmira

Farm (this study)

Period of

record

December 2019 | Volume 13 | Number 2 | e203

Herpetofauna of Bicuar National Park, Angola

Table 1 (continued). Amphibians recorded from inside and the surroundings of Bicuar National Park, Angola, based on historical

and recent records. Type of record: C = advertisement call; L = literature; O = observation; P = photograph; RR = new record for

the region; V = voucher. Period of record: A = after 2008; B = before 1975. Taxonomy has been updated over the years, therefore

original species citations may occur under different names.

Rrecics Type of Recordsinthe regionof BNP Inside Period of

P record Locality (Reference) BNP? record

BNP (Butler et al. 2019;

19. Tomopterna ahli (Deckert, 1938) Channing and Becker 2019) iG A

Catequero (Boulenger 1907);

, Calequero (= Catequero), 2 km

20. Tomopterna cf. cryptotis (Boulenger, 1907) Celery NW of (Poynton and Haacke ¥ A,B

1993); BNP (this study)

21. Tomopterna tuberculosa (Boulenger, 1882) BNP (Butler et al. 2019) 4 A

32 snakes, four testudines, and one crocodilian, Table 2).

Many of Haacke’s data represent new records for the

region (see Table 2), highlighting the relevance of his

collections. The recent records resulting from this study

recorded a total of 53 taxa; 16 amphibian taxa distributed

across nine families and 13 genera (Table 1), and 36

reptilian taxa (15 lizards, 18 snakes, two testudines, and

one crocodilian) distributed among 17 families and 35

genera (Table 2). This study revealed the first record

of Xenocalamus bicolor bicolor, the second records of

Sclerophrys poweri and of Amblyodipsas ventrimaculata

(together with the record of Butler et al. [2019]), and

the fourth record of Monopeltis infuscata in Angola. In

total, 89 specimens were collected and deposited in the

herpetological collection of ISCED-Huila.

Comments on the recent records from this study

are provided in the species accounts below, which are

arranged alphabetically by class, family, genus, and

species, with notes on taxonomy and the relevance

of the discoveries. For each species, the material used

is described as the type of record, the field number of

the collected specimen when applicable, and a code in

brackets which represents the site where the specimen

was recorded (see Appendix 1). Some records could

not be assigned to a species with certainty because no

voucher specimens were available for verification (e.g.,

Hildebrandtia, Afrotyphlops), or due to taxonomical

uncertainty in the group to which they belong (e.g.,

Hyperolius, Hemisus, Panaspis). In these cases, the

nomenclature followed Sigovini et al. (2016). Collected

tadpoles were not identified and are therefore not listed

as materials below, and their future identification awaits

the results of DNA sequencing. The only exceptions to

this exclusion are the tadpoles assigned to Kassina.

Species Accounts

Amphibia

Arthroleptidae

Leptopelis bocagii (Gunther, 1865)

Bocage's Burrowing Treefrog (Fig. 4A—B)

Amphib. Reptile Conserv.

Material: NB520 (30); NB546 (28); NB547 (29);

NB710 (27); NB765 (27); NB766 (27).

Comment: Reported in several localities in Angola

(Ceriaco et al. 2018c; Marques et al. 2018), including

recent records from Huila Province (Baptista et al. 2018).

It is considered a complex of cryptic species (Schiotz

1999), and the Angolan material assigned to Hylambates

angolensis Bocage, 1893, currently in the synonymy of

L. bocagii, requires further study (Perret 1976). Dorsal

coloration of individuals in BNP varied from completely

La]

= . % , r ‘ * ‘ad a } + : is LA ’ “ of tye ne st

Fig. 4. Leptopelis bocagii, BNP. (A) female with plain back;

(B) male with blotch in back.

December 2019 | Volume 13 | Number 2 | e203

Baptista et al.

plain to black with a horseshoe shaped blotch with white

dots in the back (Fig. 4A—B).

Bufonidae

Sclerophrys poweri (Hewitt, 1935)

Western Olive Toad (Fig. 5)

Material: NB512 (36); NB756 (25); NB764 (26);

NB767 (26); NB768 (26); NB769 (26).

Comment: Also occurring in northern Namibia and

Botswana (Channing 2001), this is the second confirmed

record of the species for Angola, after a previous record

near Calai (Conradie et al. 2016). Some early records

of Bufo regularis sensu latu may also refer to S. poweri

(Ruas 1996). Bufo regularis humbensis Monard, 1937

was originally described from Mulondo, close to BNP.

It is currently placed under the synonymy of S. garmani

(Tandy and Keith 1972), which in Angola refers to

S. poweri, and this synonymy should be reviewed.

Sclerophrys poweri is very abundant in BNP and was

found hundreds of meters away from water on moist

nights. Breeding was observed in permanent pans, with

males calling in choruses at night and eggs typically laid

in gelatinous single strings.

Sclerophrys pusilla (Mertens, 1937)

Southern Flat-backed Toad (Fig. 6)

Material: NB056 (31); NB763 (26).

Comment: Taxonomy and identification of bufonids

in Angola remains problematic (Baptista et al. 2019).

Sclerophrys pusilla represents populations previously

assigned to S. maculata in eastern and southern Africa,

including Angola (Poynton et al. 2016). This species is

known from several localities in the country (Poynton

and Haacke 1993; Ruas 1996, 2002; Conradie et al.

2016; Ceriaco et al. 2018c; Marques et al. 2018). It

was heard calling in BNP on 24 January 2009, but not

collected (A. Channing, pers. comm.), and was recently

recorded from Capelongo, near BNP (Butler et al. 2019).

It was also found around the Main Camp facilities and in

a permanent water body.

Hemisotidae

Hemisus cf. guineensis Cope, 1865

Guinea Snout-burrower (Fig. 7A—C)

Material: NB511 (15); NB550 (T2); NB551 (T2).

Comment: Tadpoles of this species were collected

in BNP during the 2009 survey (A. Channing, pers.

comm.). The taxonomy of Hemisus guineensis and H.

marmoratus is not fully resolved. Angolan specimens of

both species have been treated as a single taxon in the

past (Hemisus guineensis microps Laurent, 1972) and

records of the genus are scattered throughout the country

(Ruas 1996; Marques et al. 2018). Recent records of

Hemisus from Cangandala have been considered as H.

guineensis (Ceriaco et al. 2018c; Vaz Pinto and Baptista,

Amphib. Reptile Conserv.

unpub. data). Coloration of adult frogs from BNP ranges

from finely spotted forming lines, and mottled forming

continuous blotches to almost plain with very small

spots (see Fig. 7A—C). A similar pattern of variation in

coloration was reported by W. Conradie (pers. comm.)

in southeastern Angola, with all forms genetically

confirmed as being H. cf. guineensis. Based on this, and

the proximity to a previous record from the Cubango

basin (Monard 1937a), BNP specimens likely belong to

the same taxon.

Hyperoliidae

Hyperolius angolensis Steindachner, 1867 complex

Angola Reed Frog

Material: NB523 (41); NB524 (41); NB538 (27);

NB539 (27); NB540 (27); NB541 (27).

Comment: This species was heard calling in BNP (site

31) on 24 January 2009, but no material was collected

(A. Channing, pers. comm.). It was recently recorded

in BNP by Butler et al. (2019), who provided photos of

coloration variations. This is another unresolved complex

of reed frogs in Africa (see Schigtz 1999), with a number

of names available for Angolan populations (Baptista et

al. 2019, as H. parallelus). In Angola, this complex is

recorded throughout the country (Monard 1937a; Laurent

1950, 1954, 1964; Poynton and Haacke 1993; Conradie

et al. 2016; Baptista et al. 2018; Ceriaco et al. 2018c;

Marques et al. 2018), with consistent regional color

patterns. Poynton and Haacke (1993) referred to previous

records in Huila Province as Hyperolius marmoratus

huillensis.

Hyperolius benguellensis (Bocage, 1893) complex

Benguela Long Reed Frog (Fig. 8A—B)

Material: NB510 (36); NB526 (41); NB542 (27);

NB543 (27); NB544 (27).

ZMB 77273; ZMB 77274 (collected by Alan Channing,

not analyzed).

Comment: Hyperolius benguellensis belongs to the

challenging Hyperolius nasutus complex, with a

problematic taxonomy throughout Africa (Schiotz 1999;

Amiet 2005; Marques et al. 2018). Channing et al.

(2013) proposed a rearrangement for this group based on

morphology, genetics, and advertisement calls, resulting

in four species occurring in Angola: H. nasutus, H.

dartevellei, H. adspersus, and H. benguellensis. Of these,

three were originally described from Angola: Hyperolius

nasutus Gunther, 1865, type locality: Duque de Braganca

(= Calandula); HAyperolius adspersus Peters, 1877,

type locality: Chinchoxo, in Cabinda; and Hyperolius

benguellensis (Bocage, 1893) type locality: "Cahata" that

has been incorrectly assigned to Caota (e.g., Channing

2001; Marques et al. 2018; Frost 2019). In fact, Cahata

is located 5 km east of Balombo Municipality and was

a known collecting site for the famous naturalist José

de Anchieta in the nineteenth century (Bocage 1895).

December 2019 | Volume 13 | Number 2 | e203

Herpetofauna of Bicuar National Park, Angola

vad ‘ pty.

aS,

a 2 tle. rly

rophrys pusilla, BNP

Sele

ae .

Fig. 6.

Although the region is still located in Benguela Province,

it lies on the plateau at 1,230 m asl, more than 400 km

east of the coastal town of Benguela and closer to the

town of Huambo, while Caota is found on the outskirts

of Benguela at 20 m asl. The name “benguellensis” is

therefore misleading, and is probably the reason why

Cahata was confused with Caota, but the latter is a beach

site in the semi-arid Angolan southwest, and an unlikely

habitat for a reed frog. We therefore re-establish the type

locality of H. benguellensis to Cahata, near Balombo. The

assignment of erroneous names to Angolan localities has

been detected in other cases (Branch et al. 2017, 2018;

Vaz Pinto et al. 2019), thus special attention must be given

to this when consulting historical and recent literature.

Channing et al. (2013) assigned one specimen from BNP

to H. benguellensis, and we assign these specimens to this

name, with the altitude similar to the true type locality

providing further support. Butler et al. (2019) collected

one specimen from BNP, recording it as H. cf. nasutus,

and in this study we tentatively regard this record as

the same as ours. Both species, H. benguellensis and H.

nasutus, are sympatric in Cangandala National Park (Vaz

Pinto, unpub. data). Specimens from this group have been

also found in western Zambia (Bittencourt-Silva 2019)

and assigned to H. dartevellei and H. nasicus. Similar to

H. angolensis complex, a more complete understanding

of this complex in Angola requires countrywide surveys

and an integrated analysis of molecular, morphological,

and advertisement call data.

Kassina senegalensis (Dumeéril and Bibron, 1841)

Bubbling Kassina

Material: NB525 (T1); NB536 (tadpoles) (7); NB545

(T3); NB552 (T2); NB761 (26).

Comment: Widespread throughout Africa and in Angola

(Baptista et al. 2018; Marques et al. 2018). This species

has been previously recorded in BNP as tadpoles (A.

Channing, pers. comm.). Although subspecies have

been proposed based on color patterns (Laurent 1957),

more comprehensive studies are required to resolve this

species’ taxonomy.

- Saeed 4 ~ 3 Pr = ss im) 7 a oes!

Fig. 7. Hemisus cf. guineensis, BNP. (A—C) Three variations

in coloration.

Amphib. Reptile Conserv. 104 December 2019 | Volume 13 | Number 2 | e203

Baptista et al.

Fig. 8. Hyperolius benguellensis complex, BNP. (A) Ventral

view; (B) dorsal view.

Microhylidae

Phrynomantis bifasciatus (Smith, 1847)

Banded Rubber Frog

Comment: This species was heard calling in BNP (site

31) on 24 January 2009, but no material was collected

(A. Channing, pers. comm. ). In Angola, it is known from

Mulondo (Monard 1937a), which is located near the

boundary of BNP, in addition to Quissange and Benguela

(Bocage 1895), and Chingo (Ferreira 1904).

Phrynobatrachidae

Phrynobatrachus mababiensis FitzSimons, 1932

Mababe Puddle Frog

Comment: This species was heard calling in BNP

(site 31) on 24 January 2009, and a single tadpole (A.

Channing, pers. comm.) and adults (Butler et al. 2019)

have been collected in BNP. In Angola, it is recorded

from Lagoa Nuntechite (Poynton and Haacke 1993),

Cubango, Cuito and Cuando rivers basins (Conradie

et al. 2016), and Tundavala (Baptista et al. 2018). This

species belongs to the P. cryptotis group (Marques et al.

2018) and is in need of further taxonomic studies all over

Africa (Zimkus et al. 2010).

Amphib. Reptile Conserv.

Phrynobatrachus natalensis (Smith, 1849)

Snoring Puddle Frog

Comment: This species was heard, but not recorded,

calling in BNP (site 31) on 24 January 2009; no

material was collected (A. Channing, pers. comm.). It is

widespread in Angola (Ruas 1996, 2002; Marques et al.

2018) and includes several cryptic taxa (Zimkus et al.

2010) that are presently being investigated. Additional

specimens collected from BNP will be crucial for species

confirmation.

Pipidae

Xenopus petersii Bocage, 1895

Peters' Clawed Frog

Material: NB757 (27); NB758 (27); NB759 (27).

Comment: This species is widespread in Angola

(Monard 1937a as_X. laevis, Ruas 1996, 2002; Baptista

et al. 2018; Ceriaco et al. 2018c; Marques et al. 2018). It

has been previously recorded in BNP from both tadpoles

(A. Channing, pers. comm.) and adults (Butler et al.

2019). Furman et al. (2015) consider that in Angola,

X. petersii is widespread and X. poweri is restricted to

southeastern Angola, but few samples from Angola were

used in their analysis and this genus is worthy of a more

comprehensive assessment in the country. Hamerkop

(Scopus umbretta) and Lilac-breasted Roller (Coracias

caudatus) were seen preying upon this species in BNP.

Ptychadenidae

Hildebrandtia cf. ornata (Peters, 1878)

Ornate Frog (Fig. 9)

Material: Photographic record (F. Maiato, on wetland

between Chibemba and Cahama, approximate

coordinates same as site 42).

Comment: Two species of Hildebrandtia exist in Angola:

H. ornata and H. ornatissima (Marques et al. 2018;

Baptista et al. 2019). Hildebrantia ornata is limited to the

southwest, and H. ornatissima extends northwards to the

central plateau (Ruas 1996; Marques etal. 2018). Boulenger

(1919) provides morphological distinction between H.

ornatissima (Bocage, 1897), endemic to Angola, and H.

ornata (Peters, 1878) (= R. ruddi), originally described

from Kenya but with a wide distribution in Africa. Perret

(1976) considered H. ornata and H. ornatissima as two

valid species based on morphology, but Poynton and

Haacke (1993) contest that, and consider them subspecies.”

Specimens collected from Dongue, 28 km west of BNP

and other localities in southwestern Angola had mixed

features from both taxa and were assigned to H. ornata

ornata (Poynton and Haacke 1993). A single frog was

photographed, approximately 55 km southwest of BNP’s

boundaries, and it is provisionally assigned to H. ornata

based on the identification of material collected in close

proximity (Poynton and Haacke 1993). Hildebrandtia

December 2019 | Volume 13 | Number 2 | e203

Herpetofauna of Bicuar National Park, Angola

Pris.

ta, between Chibemba_ and

‘A (

Fig. 9. Hildebrandtia cf. orn

Cahama (Photo by F- Maiato).

specimens have also been recently collected from several

localities in Huila and Malanje provinces (Baptista and

Vaz Pinto, unpub. data), and the genus requires a revision

in the country.

Ptychadena oxyrhynchus (Smith, 1849)

Sharp-nosed Ridged Frog (Fig. 1OA—B)

Material: NB750 (25).

Comment: Species identification for Ptychadena in

Angola is not resolved (Baptista et al. 2019). A single

specimen was found near Lagoa da Matemba, but no

advertisement call was heard. It was assigned to P.

oxyrhynchus based on morphology and coloration (Fig.

10A-B). This species has recently been recorded from

BNP (Butler et al. 2019) and is known from several

localities in Angola (Marques et al. 2018).

Ptychadena porosissima (Steindachner, 1867)

Grassland Ridged Frog

Comment: This species was heard calling in BNP (site 31)

on 24 January 2009, but no material was collected and no

call was recorded (A. Channing, pers. comm.). Widespread

in sub-Saharan Africa, and recorded in Angola from the

west and the northeast (Ruas 1996; Marques et al. 2018).

Additional specimens will be important for confirmation

of species identification in BNP.

Pyxicephalidae

Pyxicephalus adspersus Tschudi, 1838

African Bullfrog

Material: Interviews.

Comment: Staff from Carmira Farm mentioned that

this frog—which is unmistakable by its distinctive size,

morphology, and behavior—appears during the first

Amphib. Reptile Conserv.

heavy rains. Pyxicephalus adspersus is recorded from

southern Angola in Humbe (Bocage 1895), Mupanda

(Monard 1937a), and Pereira d’E¢a (= Ondjiva) and 23

km NW Pereira d’Eca (Poynton and Haacke 1993, as P.

a. edulis). Several synonymies exist within P. adspersus

(Frost 2019). Consensus regarding whether P. edulis

occurs in the Zambezi Region (previously the Caprivi

Strip), has not been reached (Herrmann and Branch

2013), and the Angolan material requires further analysis.

Locally called “mafuma,” it 1s captured for consumption

and bushmeat trade. It may also have cultural relevance

in Angola, as in some parts of Cunene Province this

frog’s mating behavior is said to inspire a traditional fight

called “engolo” among the Nkhumbi people (J. Moniz,

pers. comm. ).

Tomopterna cf. cryptotis (Boulenger, 1907)

Cryptic Sand Frog (Fig. 11 A—C)

Material: NB751 (25); NB752 (25); NB753 (25);

NB754 (25); NB755 (25); NB762 (26).

Comment: The species was found breeding in early

December, with males calling in loud choruses on

banks of water bodies in BNP. Collected specimens

had considerable coloration variation (see Fig. 11A—C).

Tomopterna cryptotis was originally described from

Catequero (ca. 95 km south of BNP), and has been

recorded between Calequero (= Catequero) and Cahama

(ca. 75 km southwest of BNP) [Poynton and Haacke

1993] among other localities in Angola (Ruas 1996;

Marques et al. 2018). This species is morphologically

indistinguishable from 7’ tandyi (Channing and Bogart

1996), which was originally described from the Eastern

Cape in South Africa and is known from Namibia,

Botswana, and South Africa (Channing 2001, du Preez

and Carruthers 2009). Although 7° tandyi is considered

as being present in southwestern Angola (Channing 2001;

Channing and Rodel 2019), there are no literature records

from the country, and its occurrence is presumably based

on the morphology of similar frogs in Namibia (A.

Channing, pers. comm.). However, 7Zomopterna species

are highly cryptic, and difficult to distinguish based on

morphology. Due to the fact that the BNP records are near-

topotypical, these specimens are assigned to 7’ cryptotis,

but the advertisemen calls heard in BNP resembled

those of 7? tandyi provided by du Preez and Carruthers

(2019). Further integrative revision of these species in

Angola is needed to confirm this assignment. Two other

species from the same genus have recently been recorded

from BNP (Butler et al. 2019): 7 tuberculosa and T.

damarensis, which was recently re-assigned to 7ompterna

ahli (Channing and Becker 2019), which means that at

least three species may occur sympatrically. Given

the highly cryptic morphology of species within this

genus (Channing and Rodel 2019), and the continuing

descriptions of new species (Wilson and Channing 2019),

all of these new records require revision.

December 2019 | Volume 13 | Number 2 | e203

Baptista et al.

Fig. 10. P

detail of thighs.

Reptilia

Squamata

Sauria

Agamidae

Acanthocercus sp.

Tree Agama (Fig. 12)

Material: NB709 (31).

Comment: This species was previously assigned to

Acanthocercus atricollis (Smith, 1849) until Wagner

et al. (2018) revived the name A. cyanocephalus for

western populations, which included Angola. Reported

in several publications from Angola (Bocage 1879, 1895;

Ferreira 1902; Boulenger 1905; Monard 1931, 1937b;

Schmidt 1933; Laurent 1950, 1964; Manacas 1963;

Conradie et al. 2016; Ceriaco et al. 2018c; Marques et

al. 2018), but material from Huila Province belongs

to a different species, which is in the process of being

described (Marques et al. 2018; Butler et al. 2019).

Recently recorded in BNP (Butler et al. 2019), abundant

at the park’s headquarters, and found on large and tall

Burkea africana trees.

Agama aculeata (Merrem, 1820)

Ground Agama

Material: NB770 (32).

Comment: Ground Agamas occur widely in southern

Africa (Bates et al. 2014) and are reported throughout

Angola (Marques et al. 2018). The taxonomy of Angolan

populations remains unresolved, and a recent phylogeny

of Agama (Leaché et al. 2014) did not include Angolan

Amphib. Reptile Conserv.

ra P B > © Te

a Ce a ee

Fig. 11. Zomopterna cf. cryptotis, BNP. (A—C) Three variations

in coloration.

samples. Recently recorded in BNP (Butler et al. 2019),

in this study it was found on the ground in degraded

shrubland in the outskirts of BNP.

Amphisbaenidae

Monopeltis cf. anchietae (Bocage, 1873)

Anchieta's Spade-snouted Worm Lizard (Fig. 13)

Material: One photographic record of a freshly killed

individual (L. Gata, CF).

December 2019 | Volume 13 | Number 2 | e203

Herpetofauna of Bicuar National Park, Angola

Table 2. Reptiles recorded from inside and the surroundings of Bicuar National Park, Angola, based on historical and recent records.

Type of record: CR = new record for the country; H = unpublished record from Ditsong (= Transvaal) Museum’s collection; L =

literature; O = observation; P = photograph; RR = new record for the region; V = voucher; E = endemic. Period of record: A =

after 2008; B = before 1975. Taxonomy has been updated over the years, therefore original species nomenclature may occur under

different names.

10.

11.

12:

13.

14.

13:

Taxa

Sauria

Agamidae

Acanthocercus sp.

Agama aculeata (Merrem, 1820)

Agama planiceps shacki Mertens, 1938

Amphisbaenidae

Monopeltis cf. anchietae (Bocage, 1873)

Monopeltis infuscata Broadley, 1997

Monopeltis perplexus Gans, 1976

Zygaspis quadrifrons (Peters, 1862)

Chamaeleonidae

Chamaeleo dilepis quilensis (Bocage

1886)

Cordylidae

Cordylus machadoi Laurent, 1964

Gekkonidae

Chondrodactylus fitzsimonsi (Loveridge,

1947)

Chondrodactylus laevigatus (Fischer,

1888)

Hemidactylus benguellensis Bocage, 1893

Lygodactylus angolensis Bocage, 1896

Lygodactylus bradfieldi Hewitt, 1932

Pachydactylus punctatus Peters, 1854

complex

Amphib. Reptile Conserv.

Endemic?

Type of

record

mbt Bs

H,L,V

LEN:

108

Records in the region

of BNP Inside Period of

Locality (and BNP? record

Reference)

Mupa (Monard 1937b);

BNP (Butler et al. 2019; Yi A,B

this study)

Cahama and Chibemba,

21 km NW of (Haacke,

TM); BNP (Butler et al. y

2019; this study)

Chibemba, 5 km S;

Dongue, 10 km NW of A,B

(Haacke, TM)

Humbe (Bocage 1873);

Mupa (Monard 1937b); A,B

Carmira Farm (this study)

Humbe (Bocage 1873;

Broadley 1997); Carmira A,B

Farm (this study)

Humbe (Gans 1976) B

BNP (Butler et al. 2019);

Carmira Farm (this study) :

Cahama, 23 km SE

(Haacke, TM); Kului

(Monard 1937b); BNP Y A,B

(Butler et al. 2019; this

study)

Chibemba, 5 km S;

Humbia, 12 km E B

(Haacke, TM)

Viriambundo (Haacke,

TM)

Mulondo (Schmidt 1933);

Humbe (Monard 1937b);

BNP (Butler et al. 2019); ;

Carmira Farm (this study)

Capelongo (Butler et al.

2019)

BNP (this study) Y

Humbe-Cahama, 36 km

from; Humbe, 5 km N of;

Humbia (Haacke, TM); é

BNP (Butler et al. 2019)

Kului (Monard 1937b;

Bauer et al. 2006); BNP yy A,B

(this study)

December 2019 | Volume 13 | Number 2 | e203

Baptista et al.

Table 2 (continued). Reptiles recorded from inside and the surroundings of Bicuar National Park, Angola, based on historical and

recent records. Type of record: CR = new record for the country; H = unpublished record from Ditsong (= Transvaal) Museum’s

collection; L = literature; O = observation; P = photograph; RR = new record for the region; V = voucher; E = endemic. Period of

record: A = after 2008; B = before 1975. Taxonomy has been updated over the years, therefore original species nomenclature may

occur under different names.

Records in the region

; Type of of BNP Inside Period of

US pens record Locality (and BNP? record

Reference)

Rhoptropus barnardi Hewitt, 1962 eek a eee

He: H, RR Humbia, 12 km E B

(Haacke, TM)

Gerrhosauridae

Capelongo, Mulondo

(Monard 1937b);

Gerrhosaurus bulsi-multilineatus complex pees Bs ae Pee

17. E* Hale Weck es ee Y A,B

me ¥. Chibemba, Cahama, 30 °

km SE of (Haacke, TM);

BNP (Butler et al. 2019;

this study)

Dongue, 10 km NW of

18. Matobosaurus maltzahni (De Grys, 1938) H, RR (Haacke, TM) B

Lacertidae

Mulondo (Monard

19. Heliobolus lugubris (Smith, 1838) H, L 1937b); Cahama, 3 km B

NW of (Haacke, TM)

20. Ichnotropis bivittata Bocage, 1866 L Kului (Monard 1937b)

BNP (Butler et al. 2019;

21. Ichnotropis capensis (Smith, 1838) LIV this study) A

22. Meroles squamulosus (Peters, 1854) |B CapelongoMonard B

1937b)

3 Nucras broadleyi Branch, Conradie, Vaz E L Capelongo (Monard B

’ Pinto, Tolley 2019 1937b)

Scincidae

24. Acontias occidentalis FitzSimons, 1941 L pipe (Monaro:

FitzSimons 1941)

25. Mochlus sundevalli (Smith, 1849) RR, V_— Carmira Farm (this study)

; a Bare BNP (Butler et al. 2019;

26. Panaspis wahlbergi-maculicollis complex | es this study) nA

Chibemba (Laurent

27. Sepsina angolensis Bocage, 1866 | ae 1964); Viriambundo B

(Haacke, TM)

28. Trachylepis albopunctata (Bocage, 1867) L BNP (Butler et al. 2019) Y A

Mupa (Monard 1937b);

ae: Chibemba, 11 km S;

29. Trachylepis binotata (Bocage, 1867) Hi Ts Humbe (Haacke, TM): wg A,B

BNP (Butler et al. 2019)

Humbe (Schmidt 1933);

30. Trachylepis chimbana (Boulenger, 1887) H Humbia, 12 km E of B

(Haacke, TM)

Humbe to Cahama, 36

PyOA 2e km NW of (Haacke,

31. Trachylepis spilogaster (Peters, 1882) H, L, V TM): BNP (Butler et al. Y A,B

2019; this study)

Trachylepis sulcata ansorgii (Boulenger, Chibemba, 11 km S

32 4907) H, RR (Haacke, TM) :

Amphib. Reptile Conserv. 109 December 2019 | Volume 13 | Number 2 | e203

Table 2 (continued). Reptiles recorded from inside and the surroundings of Bicuar National Park, Angola, based on historical and

recent records. Type of record: CR = new record for the country; H = unpublished record from Ditsong (= Transvaal) Museum’s

collection; L = literature; O = observation; P = photograph; RR = new record for the region; V = voucher; E = endemic. Period of

record: A = after 2008; B = before 1975. Taxonomy has been updated over the years, therefore original species nomenclature may

Herpetofauna of Bicuar National Park, Angola

occur under different names.

ao!

34.

oot

36.

aM.

38.

39.

40.

4].

42.

43,

44,

45.

46.

Taxa

Trachylepis sulcata sulcata (Peters, 1867)

Trachylepis varia (Peters, 1867) clade B

(Weinell and Bauer 2018)

Trachylepis wahlbergi (Peters, 1869)

Varanidae

Varanus albigularis angolensis Schmidt,

1933

Serpentes

Colubridae

Crotaphopeltis hotamboeia (Laurenti,

1768)

Dasypeltis scabra (Linnaeus, 1758)

Dispholidus typus viridis (Smith, 1838)

Philothamnus angolensis Bocage, 1882

Philothamnus semivariegatus (Smith,

1840) sensu lato

Telescopus semiannulatus polystictus

Mertens, 1954

Thelotornis capensis oatesi (Gunther,

1881)

Elapidae

Dendroaspis polylepis Gunther, 1864

Elapsoidea semiannulata semiannulata

Bocage, 1882

Naja anchietae Bocage, 1879

Amphib. Reptile Conserv.

Type of

record

110

L,V

LV.

Records in the region

of BNP

Locality (and

Reference)

Capelongo (Butler et al.

2019)

Capelongo (Monard

1937b); Cahama, 21

km NW — Chibemba,

Humbia, 12 km E of

(Haacke, TM)

Humbi, Capelongo,

Kulu, Mulondo, Mupa

(Monard 1937b); Humbe,

5 km N of (Haacke, TM)

Mulondo, Mupa (Monard

1937b); BNP (this study)

Gambos, Humbe (Bocage

1895); Capelongo

(Monard 1937b);

Nougalafa Lake (this

study)

Gambos (Bocage 1895)

Humbe (Bocage 1895);

Mupa (Monard 1937b);

Capelongo (Bogert

1940); Carmira Farm

(this study)

Capelongo (Bogert

1940); Humbe (Haacke,

TM)

Humbe (Bocage 1895);

Mupa (Monard 1937b),;;

Carmira Farm, Handa

Farm (this study)

Humbe, Gambos (Bocage

1895)

BNP (this study)

Mulondo (Schmidt

1933); BNP, Handa Farm

(this study)

Gambos (Bocage 1895;

Broadley 1998)

Humbe (Bocage 1895);

Capelongo (Bogert

1940); Mupa (Monard

1937b); BNP, Handa

Farm (this study)

December 2019 | Volume 13 | Number 2 | e203

Period of

record

)

Baptista et al.

Table 2 (continued). Reptiles recorded from inside and the surroundings of Bicuar National Park, Angola, based on historical and

recent records. Type of record: CR = new record for the country; H = unpublished record from Ditsong (= Transvaal) Museum’s

collection; L = literature; O = observation; P = photograph; RR = new record for the region; V = voucher; E = endemic. Period of

record: A = after 2008; B = before 1975. Taxonomy has been updated over the years, therefore original species nomenclature may

occur under different names.

47.

48.

49.

50.

51.

a2:

53.

54.

53:

56.

57.

58.

bee

60.

61.

62.

63.

Taxa

Naja nigricollis Reinhardt, 1843

Lamprophiidae

Amblyodipsas polylepis (Bocage, 1873)

Amblyodipsas ventrimaculata (Roux,

1907)

Aparallactus capensis Smith, 1849

Boaedon angolensis Bocage, 1895

Hemirhagerrhis viperina (Bocage, 1873)

Prosymna angolensis Boulenger, 1915

Prosymna visseri Fitzsimons, 1959

Psammophis angolensis (Bocage, 1872)

Psammophis mossambicus Peters, 1882

Psammophis subtaeniatus Peters, 1882

Psammophylax tritaeniatus (Gunther,

1868)

Psammophylax ocellatus (Bocage, 1873)

Pseudaspis cana (Linnaeus, 1758)

Xenocalamus bicolor bicolor Gunther,

1868

Leptotyphlopidae

Leptotyphlops scutifrons (Peters, 1854)

Namibiana aff. rostrata (Bocage, 1886)

Amphib. Reptile Conserv.

Type of

record

| gf

L,PV

L,V

111

Records in the region

of BNP

Locality (and

Reference)

Humbe (Bocage 1895);

Capelongo (Bogert

1940); Osi (= Osse)

(Monard 1937b); Handa

Farm (this study)

Humbe (Bocage 1895)

BNP (Butler et al. 2019;

this study)

Gambos (Bocage 1895)

Cahama, 3 km NW of

(Haacke, TM); BNP

(Butler et al. 2019);

Carmira Farm (this study)

Humbe (Bocage 1895);

Chibemba, 5 km S

(Haacke, TM)

Capelongo (Bogert

1940); BNP (this study)

Chibemba, 5 km S

(Haacke, TM)

Humbe (Bocage 1895;

Schmidt 1933)

Capelongo (Bogert

1940); Mupa (Monard

1937b); BNP, Handa

Farm (this study)

Humbe, Mulondo, Mupa

(Monard 1937b); BNP

(Butler et al. 2019);

Carmira Farm (this study)

Gambos, Humbe

(Monard 1937b),;

Capelongo (Bogert

1940); Humbe—Cahama,

36 km NW of (Hacke,

TM)

Gambos, Humbe (Bocage

1895; Branch et al.

2019a)

BNP (Butler et al. 2019);

Carmira Farm (this study)

Capelongo (Monard

1937b); Quipungo

(Haacke, TM)

Humbe (Bocage 1895);

BNP (Butler et al. 2019);

Handa Farm (this study)

Inside Period of

BNP? record

A,B

ne A

Y A,B

yy B

Y B

Y: A,B

A,B

Ne A

Y A,B

December 2019 | Volume 13 | Number 2 | e203

Herpetofauna of Bicuar National Park, Angola

Table 2 (continued). Reptiles recorded from inside and the surroundings of Bicuar National Park, Angola, based on historical and

recent records. Type of record: CR = new record for the country; H = unpublished record from Ditsong (= Transvaal) Museum’s

collection; L = literature; O = observation; P = photograph; RR = new record for the region; V = voucher; E = endemic. Period of

record: A = after 2008; B = before 1975. Taxonomy has been updated over the years, therefore original species nomenclature may

occur under different names.

Taxa Endemic?

Pythonidae

64. Python anchietae Bocage, 1887

65. Python natalensis Smith, 1840

Typhlopidae

Afrotyphlops cf. schlegelii (Bianconi,

1849)

Viperidae

66.

67. Bitis arietans Merrem, 1820

68. Causus rhombeatus (Lichtenstein, 1823)

Testudines

Pelomedusidae

69. Pelomedusa subrufa (Bonnaterre, 1789)

70. Pelusios cf. nanus Laurent, 1956

Testudinidae

71. Kinixys cf. belliana Gray, 1831

72. Stigmochelys pardalis (Bell, 1828)

Crocodilia

Crocodylidae

73. Crocodylus niloticus Laurenti, 1768

Comment: Monopeltis anchietae is recorded from

southwestern Angola (Marques et al. 2018), and Branch

et al. (2019b) discuss its taxonomical history. The

coloration pattern of the specimen (Fig. 13) conforms to

the most common pattern associated with M. anchietae

(Broadley et al. 1976), but the photograph resolution

does not allow for the detailed scale counts necessary

to provide a positive identification. The species was

recorded in Humbe (Bocage 1873), approximately 110

km south of BNP, and the Carmira Farm specimen is

provisionally assigned to M. anchietae, pending the

collection of additional material for confirmation.

Monopeltis infuscata Broadley, 1997

Infuscate Spade-snouted Worm Lizard (Fig. 14)

Material: One unlabelled and bleached specimen in

Carmira Farm’s private collection.

Amphib. Reptile Conserv.

TAZ

Records in the region

Type of of BNP Inside Period of

record Locality (and BNP? record

Reference)

Viriambundo (Haacke,

H, RR TM) B

Capelongo (Bogert

L,V 1940); BNP (this study) my a

Humbe (Bocage 1893);

L,P BNP (this study) ae

Capelongo (Bogert

L,V 1940); BNP (this study) si a B

P,RR ~~ Handa Farm (this study) A

Humbe (Bocage 1895);

L,V BNP (Butler et al. 2019; Y A,B

this study)

L Osi (= Osse) (Monard B

1937b); Broadley (1981)

Osi (= Osse) (Monard

La 1937b); BNP (Butler et ¥ A,B

al. 2019; this study)

ie Mupa (Monard 1937b) B

L.O Capelongo (Monard AB

1937b); BNP (this study) i

Comment: Scalation features of the specimen (dorsal

head shield with blind lateral sutures (Fig. 14), four

postgenials in the first row, more than seven postgenials

in the second row, two precloacal pores, 204 body

annuli, and 10 caudal annuli) match the description of

M. infuscata (Broadley 1997). In Angola, the species

was recorded in Humbe in Cunene Province, Tombole

River in Cuando-Cubango Province, and a locality

named “Sturuba” (Broadley 1997) which could not be

determined. This is the third confirmed locality for the

Species in the country and the first for Huila Province,

and additional voucher specimens and genetic material

should be collected. Found among sandy soils in Baikea/

Burkea woodlands, Monopeltis anchietae and M.

infuscata are known to be sympatric in Humbe (Broadley

1997), approximately 110 km south of BNP, supporting

their co-occurrence in Carmira Farm.

December 2019 | Volume 13 | Number 2 | e203

Baptista et al.

yal

Fig. 12. Acanthocercus sp., BNP (male). |

Zygaspis quadrifrons (Peters, 1862)

Kalahari Round-snouted Worm Lizard (Fig. 15)

Material: One unlabelled and bleached specimen in

Carmira Farm’s private collection.

Comment: This specimen conforms morphologically

to Z. quadrifrons (Broadley and Broadley 1997). For

many years, the species was known from Angola only

from Monard’s (1931) description of Amphisbaena

ambuellensis from ‘Chimporo’ (= Tchimpolo), which

was subsequently synonymized with A. qguadrifrons by

Loveridge (1941) and later transferred to Zygaspis by

Alexander and Gans (1966). No additional Angolan

material was collected until recent surveys in southeastern

Angola (Conradie et al. 2016) and BNP (Butler et

x} :

> ~

A gees. ea

sft Py ar ita i ae > eS eae

Fig. 13. Monopeltis cf. anchietae, Carmira Farm (Photo by L.

Gata).

Amphib. Reptile Conserv.

al. 2019). Broadley and Broadley (1997) recognized

five groups within Z. qguadrifrons. Additional voucher

specimens and genetic material are necessary to confirm

species identification, and to evaluate the validity of

Monard’s taxon “ambuellensis” for Angolan material

(Branch et al. 2019c). Found in the same habitat as M.

infuscata.

Chamaeleonidae

Chamaeleo dilepis quilensis (Bocage, 1886)

Quilo Flap-necked Chameleon (Fig. 16)

Material: NB760 (33); NB 1084 (C1).

Comment: This subspecies is common and widespread

throughout Angola (Marques et al. 2018). It was recently

recorded in BNP (Butler et al. 2019) and observed

throughout the park, and it has been historically recorded

near BNP (Monard 1937b; Haacke, TM).

Gekkonidae

Chondrodactylus aff. laevigatus (Fischer, 1888)

Button-scaled Gecko

Comment: <A_ large individual of the genus

Chondrodactylus was observed on _ buildings at

Carmira Farm (N. Baptista, pers. obs.), but 1t was not

photographed or collected, and is provisionally assigned

to C. laevigatus based on recent records of this species

from BNP (Butler et al. 2019). Records from near the

park in Mulondo (Schmidt 1933, as Pachydactylus

stellatus), and Humbe (Monard 1937b, as P. bibroni) are

also assigned to this taxon, which is known to occur in

southwestern Angola (Marques et al. 2018).

Lygodactylus angolensis Bocage, 1896

Angola Dwarf Day Gecko (Fig. 17)

Material: NB517 (31).

Comment: Specimens of this taxon were first described

as L. capensis by Bocage (1895), who subsequently

assigned them to a new species, L. angolensis, described

Fig. 14. Monopeltis infuscata, Carmira Farm.

December 2019 | Volume 13 | Number 2 | e203

Herpetofauna of Bicuar National Park, Angola

Fig. 15. Zygaspis quadrifrons, Carmira Farm.

from Hanha, Benguela Province (Bocage 1897). Schmidt

(1933) described a new species of dwarf day gecko, L.

laurae, which was later synonymized with L. angolensis

(Marques et al. 2018; Branch et al. 2019c). Widely

distributed through Africa, localities in Angola were

compiled by Marques et al. (2018) and include Chimboa

da Hanha (= Capira); Cacondo (= Caconda), 2 km W

of; Cubal; Cutenda, 3 km S of; Negola, 6 km S of; and

Quimbango (Haacke, TM). The single BNP specimen

conforms to the original description of L. angolensis

in scalation and coloration, and is similar to specimens

collected in the urban environments of Lubango

(Baptista, unpub. data). Specimens collected at the same

localities (BNP and Lubango) on other occasions have

been assigned to L. bradfieldi (Butler et al. 2019), and

these observations deserve further morphological and

genetic comparisons.

Pachydactylus punctatus Peters, 1854 complex

Speckled Thick-toed Gecko (Fig. 18)

Material: NB513 (14); NB514 (T1); NBS15 (11);

NBS530 (T3); NB773 (34); NB774 (34); NB775 (34).

Comment: This species was recorded in southwestern

Angola (Monard 1937b; Laurent 1964; Marques et al.

2018), including Kului, near BNP (in Monard 1937b

as P. serval, assigned to P. punctatus by Bauer et al.

2006). BNP specimens are morphologically different

ae. ell

Fig. 17. Lygodactylus angolensis, BNP.

Amphib. Reptile Conserv.

Fig. 16. Chamaeleo dilepis quilensis, BNP.

(larger, sturdier, paler, and with a different blotching

pattern) from other specimens found in Tundavala and

Lubango, Huila Province (Baptista et al. 2018; Butler

et al. 2019), that are also assigned to the P punctatus

complex. These lizards also inhabit different habitats

and microhabitats (found on the ground in rocky areas

in Tundavala; under the bark of fallen logs in recently

burnt areas during the day and in leaf litter near a dirt

road during the night in BNP) and very likely belong to

different species. Tundavala is along the high altitude

edge of the Angolan plateau (above 2,000 m asl) and,

given the very specious nature of this genus (Heinicke et

al. 2017) and the existence of cryptic diversity reported

in Angola (Branch et al. 2017), the taxonomic status of

this complex requires further investigation.

Gerrhosauridae

Gerrhosaurus bulsi-multilineatus complex

Plated Lizard (Fig. 19)

Material: NB531 (31); NB776 (2); NB777 (3).

Comment: Five species of Gerrhosaurus are known to

exist in Angola: G. auritus, G. bulsi, G. multilineatus,

G. nigrolineatus, and G. skoogi (Branch et al. 2019c).

Gerrhosaurus nigrolineatus is recorded in areas adjacent

to BNP, in Capelongo and Mulondo (Monard 1937b),

and from the surroundings of Cahama (Haacke, TM).

a on ss ee ‘ .

/ is sh he

a ee Ss, ‘7 Teg .

ee ae r a

Fig. 18. Pachydactylus punctatus complex, BNP.

December 2019 | Volume 13 | Number 2 | e203

Baptista et al.

Fig. 19. Gerrhosaurus bulsi-multilineatus complex (juvenile),

BNP.

Bates et al. (2013) discussed the problematic status of

Angolan populations referred to as G. nigrolineatus,

and species boundaries within the G. bu/si-multilineatus

complex remain unresolved. Assignment of recently

collected specimens is pending until genetic assessment

of the Angolan material is published (M. Bates, pers.

comm.). Specimens collected recently in BNP by Butler

et al. (2019) were assigned to G. cf. multilineatus. One

juvenile was collected in the Main Camp (Fig. 19),

and adults were collected from burrows in degraded

shrubland near the park boundary, and they are similar to

the specimen illustrated in Butler et al. (2019).

Lacertidae

Ichnotropis capensis (Smith, 1838)

Cape Rough-scaled Lizard (Fig. 20A—B)

Material: NB771 (34); NB772 (34); NB779 (12);

photographic record (M. Finckh, site 43).

Comment: /chnotropis capensis and I. bivittata are

known to occur sympatrically in Angola (Laurent 1964;

Marques et al. 2018), and five taxa within this genus

are listed for Angola (Marques et al. 2018; Branch et

al. 2019c). The systematics of the genus /chnotropis is

poorly established (Laurent 1964), and while there is

no recent systematic revision of the group (Edwards et

al. 2013), a thorough historical revision was recently

published (Berg 2017). According to this, two subspecies

of 7. capensis occur in Angola, 1. c. capensis (Smith,

1838) and /. c. overlaeti Witte and Laurent, 1942, with

the latter being restricted to northern Angola (Marques

et al. 2018). Rough-scaled lizards have recently been

collected in BNP and were referred to IJchnotropis

bivittata pallida Laurent, 1964 (Butler et al. 2019), but

we have regarded them as /. capensis, given that J. b.

pallida is morphologically distinct, and occurs in higher

altitudes, such as Humpata (Laurent 1964). Specimens

from BNP have a bright orange/red lateral line that is more

evident in males than in females, and living males have a

bright yellow chin and chest (Fig. 20A—B) that becomes

bleached when preserved. Detailed and comprehensive

studies of species within this genus in Angola are needed.

Amphib. Reptile Conserv.

ota bee at

Fig. 20. Ichnotropis capensis, BNP. (A) Female; (B) male.

Scincidae

Mochlus sundevalli (Smith, 1849)

Sundevall's Writhing Skink (Fig. 21)

Material: One unlabelled bleached specimen in Carmira

Farm’s private collection.

Comment: Widespread throughout eastern and southern

Africa, in Angola this species is recorded from the

coastal plains south of Cuanza River, lower Cuando

River basin (records compiled in Marques et al. [2018]),

and northeastern Angola in Dundo (Laurent 1964).

Panaspis wahlbergi-maculicollis complex

Snake-eyed Skink (Fig. 22A—B)

Material: NB516 (T3); NB548 (T1); NB549 (T2).

Comment: Small leaf-litter inhabiting skinks have nu-

merous cryptic lineages in southern and eastern Africa

(Medina et al. 2016). Historically, Bocage (1895) report-

ed on material from Caconda and Cahata collected by

Anchieta, and recently P. maculicollis was recorded from

southeastern Angola (Conradie et al. 2016). A popula-

tion of “P. wahlberg7” in northern Namibia was shown to

form part of the P. maculicollis complex (Medina et al.

2016) and was subsequently described as a new species,

Panaspis namibiana (Ceriaco et al. 2018a). Snake-eyed

skinks recently collected in BNP were assigned to P. aff.

namibiana (Butler et al. 2019). BNP specimens from this

study have fused anterior parietals, conforming to the P.

wahlbergi complex, but prefrontals are well separated

(see Fig. 22B), distinguishing them from P. namibiana

(Ceriaco et al. 2018a). The taxonomic status of the BNP

December 2019 | Volume 13 | Number 2 | e203

Herpetofauna of Bicuar National Park, Angola

Fig. 21. Mochlus sundevalli, Carmira Farm.

population and its affinities to the P. maculicollis or P.

wahilbergi radiations requires further study, as do other

Angolan populations from Humpata, Quilengues, and the

Cuanza Sul escarpment (Vaz Pinto and Baptista, unpub.

data).

Trachylepis spilogaster (Peters, 1882)

Kalahari Tree Skink

Material: NB519 (31); NB527 (10); NB528 (9); NB529

(13); NB532 (6); NB533 (9); NB778 (31).

Comment: Recently recorded in BNP (Butler et al.

2019), it was the most frequently observed reptile species

during the BNP surveys, and together with Pachydactylus

puntactus, was one of only two species to be found in the

park’s woodlands after recent fires.

Varanidae

Varanus albigularis angolensis Schmidt, 1933

Angolan Savanna Monitor (Fig. 23)

Material: One photographic record (M. Finckh, site 40).

Comment: Several records of Varanus albigularis from

Angola have been assigned to two different subspecies

(Marques et al. 2018). Schmidt (1933) described V. a.

angolensis from *“Gauca, Bihe’ (= Zauca River, Malanje)

(Crawford-Cabral and Mesquitela 1989), and the BNP

record is assigned to this taxon. This species still needs

a wide-ranging phylogenetic assessment. The individual

was observed in the ecotone between grassland and

woodlands.

Serpentes

Colubridae

Crotaphopeltis hotamboeia (Laurenti, 1768)

White-lipped Herald Snake

Material: NB522 (41); four unlabelled specimens in

Carmira Farm’s private collection; one photographic

record from Handa Farm (J. Traguedo, HF).

Comment: A widespread and common species in Angola

(Marques et al. 2018), found active at night, on the edge

of Lagoa Nougalafa (= Nugarrafa, = Nongalafa).

Amphib. Reptile Conserv.

+ qs Oe"

—— = 7 pia atk. | :

Rae i

Sod, fm. © si.

fa" We elGt>-s

ie, et Mt oy"

af i ee ie i ope e

e <

ee 7

Fig. 22. Panaspis wahlbergi-maculicollis complex, BNP. Head

(A) lateral view and (B) dorsal view.

Dispholidus typus viridis (Smith, 1838)

Green Boomslang (Fig. 24)

Material: Two unlabelled bleached juvenile specimens

in Carmira Farm’s private collection; one photographic

record (L. Gata, CF).

Comment: Branch (2018) provides an overview of the

two subspecies of boomslang existing in Angola, D. t¢.

punctatus and D. t. viridis. These were shown to deserve

full species status (Eimermacher 2012), but this taxonomic

adjustment requires further consensus. Neither scalation

nor juvenile coloration allow assignment of the Carmira

Farm specimens to either taxon, but the green coloration

of a photographed adult (Fig. 24) is consistent with D. t.

viridis (Eimermacher 2012), as are other material from

Humbe. Resolution of species boundaries in Dispholidus

and the assignment of Angolan populations requires

further genetic studies.

Philothamnus semivariegatus (Smith, 1840) sensu lato

Spotted Bush Snake (Fig. 25)

Material: Two unlabelled bleached specimens in

Carmira Farm’s private collection; one photographic

record from Handa Farm (J. Traguedo, HF).

Comment: The complex taxonomic history of this

Species 1s discussed by Branch (2018). We cautiously

assign specimens from Carmira Farm to P. semivariegatus

based on scalation (three supra-labials entering orbit,

temporal arrangement 2+2, and keeled ventrals),

however, coloration of specimens from both farms did

not have any markings (atypical of P. semivariegatus).

December 2019 | Volume 13 | Number 2 | e203

Baptista et al.

Fig. 23. Varanus albigularis angolensis, BNP (Photo by M.

Finkch).

Historical records from Angola were compiled (Marques

et al. 2018), and additional records include coastal

lowlands in Benguela Province (Vaz Pinto, unpub. data).

This species is paraphyletic having at least four different

lineages (Engelbrecht et al. 2019), and the Carmira

Farm specimens might group with clade 4. Records of

P. angolensis from Capelongo (Bogert 1940) and Humbe

(Haacke, TM) deserve careful comparison with this

material.

Thelotornis capensis oatesi (Gunther, 1881)

Oates' Vine Snake (Fig. 26)

Material: NB1065 (5).

Comment: Widely distributed in Angola, with records

from Hanha (Bogert 1940), Chitado (Hellmich 1957),

Alto Chicapa (Laurent 1964), and Longa (Conradie et

al. 2016). The specimen collected in BNP conforms in

coloration and scalation to this subspecies (see Fig. 26).

Elapidae

Dendroaspis polylepis Gunther, 1864

Black Mamba

Material: Shed skin found in BNP. Photographic records

(L. Gata, CF; J. Traguedo, HF).

Comment: This species is recorded from several

Fig, 24, Disaholidus Apis viridis, Carmira aan (Photo by 2

Gata).

localities in Angola (Marques et al. 2018). Locally

known as “kuiva” (J. Traguedo, pers. comm. ).

Naja anchietae Bocage, 1879

Anchieta's Cobra (Fig. 27A—B)

Material: NB793 (21); NB250 (banded morph) [HF].

Comment: Branch (2018) discusses the description,

history, and taxonomy of N. anchietae in Angola. This

is acommon species occurring in southern Angola, with

known records compiled by Broadley (1995). Recent

records are from Malanje Province (Ceriaco et al. 2014,

Vaz Pinto, unpub. data), on the edge of the plateau in

Tundavala (Baptista et al. 2018), Cassinga (Vaz Pinto,

unpub. data), and Bimbe (Baptista and Vaz Pinto, unpub.

data).

Naja nigricollis Reinhardt, 1843

Black-necked Spitting Cobra (Fig. 28)

Material: Photographic record (J. Traguedo, HF).

Comment: The taxonomy of African cobras (Naja) has

undergone significant changes in recent years (Branch

2018). The name Naja nigricollis has a complex

history of synonymies and varieties, and only when two

varieties, N. n. nigricincta and N. n. woodi, were elevated

to full species, was the name N. nigricollis stabilized to

represent a species (Wuster et al. 2007). It is widespread

Fig. 25. Philothamnus semivariegatus sensu latu, Carmira

Farm.

Amphib. Reptile Conserv.

Fig. 26. Thelotornis capensis oatesi, BNP (Photo by T: ion

December 2019 | Volume 13 | Number 2 | e203

Herpetofauna of Bicuar National Park, Angola

Fig. Ep Naja anchietae. (A) BNP (Photo by F- Maiato); (B)

Handa Farm (Photo by J. Traguedo).

in Angola, but avoids dense forest. Locally known as

“turula n'jila” (J. Traguedo, pers. comm. ).

Lamprophiidae

Amblyodipsas ventrimaculata (Roux, 1907)

Kalahari Purple-glossed Snake

Material: NB595 (20); one individual seen in grasslands

near lake from Main Camp (site T3), not collected.

Comment: The specimen’s scalation (15 midbody scale

rows, 21 subcaudals, 203 ventrals, five upperlabials,

2™ and 3™ entering the eye) and coloration (similar to

specimen illustrated in Butler et al. [2019]) conform to

A. ventrimaculata (Branch 1998; Marais 2004). Together

with Butler et al. (2019), BNP is the second record of this

species for Angola, the first being from the Cuito River

source (Portillo et al. 2018; Branch et al. 2019c), and

represents a northwestern extension of the known range

(Botswana, Zimbabwe, Namibia, and Zambia).

Boaedon angolensis (Bocage, 1895)

Angolan House Snake (Fig. 29A—B)

Material: One unlabelled bleached specimen in the

private collection of Carmira Farm.

Comment: This species was recently recorded from BNP

(Butler et al. 2019). The scalation of the Carmira Farm

specimen (23 midbody scale rows, 116 ventrals, more than

55 subcaudals, see Fig. 29A—B for head scalation), accords

with the revision of Angolan house snakes currently in

progress (Hallermann et al., unpub. data).

Amphib. Reptile Conserv.

Ba eae EZ eS ba" eee

igricollis, Handa Farm (Photo by J. Traguedo).

Prosymna angolensis Boulenger, 1915

Angolan Shovel-snout Snake (Fig. 30)

Material: NB521 (T1).

Comment: Originally described from Huila, Angola

(discussion on type locality in Branch [2018]), this

species was recorded historically from southwestern and

central Angola (Marques et al. 2018), and more recently,

from the southeast of Angola (Conradie et al. 2017) and

Tundavala (Baptista, unpub. data). Its upper and lower

labials, ventral scales (145), and low subcaudal counts

(16) conform to the species characterization (Bocage

1895, as P. frontalis). It has gray coloration with a dark

blotch behind the head, a series of paired dark spots

along the back, and immaculate white/creamish belly and

flanks. It was found in sandy soils in miombo woodlands.

Psammophis mossambicus Peters, 1882

Olive Grass Snake (Fig. 31)

Material: NB 518 (head only) (T1); photographic record

(J. Traguedo, HF).

Comment: This species belongs to the P_ sibilans

complex (Kelly et al. 2008, Trape et al. 2019). The

collected snake had a uniformly pale-yellow belly,

and dorsal coloration was plain gray with a thin darker

vertebral dash. Head scalation and ventral (159) and

subcaudal (80) scale counts recorded from the specimen

conform to P. mossambicus (Broadley 2002, Trape et

al. 2019). It is locally known as “muiha on njolo” (J.

Traguedo, pers. comm. ).

Psammophis subtaeniatus Peters, 1882

Stripe-bellied Sand Snake (Fig. 32)

Material: Four unlabelled specimens from Carmira

Farm private collection; one photographic record (L.

Gata, CF).

Comment: This species is restricted to the semi-arid

scrubland and mopane woodland, above and below the

escarpment in southwestern Angola (Branch 2018).

Additional recent records include specimens from

Equimina and Serra da Neve (Vaz Pinto and Baptista,

unpub. data).

December 2019 | Volume 13 | Number 2 | e203

Baptista et al.

Fig. 29. Boaedon angolensis, Carmira Farm. Head (A) lateral

view and (B) dorsal view.

Pseudaspis cana (Linnaeus, 1758)

Mole Snake

Material: One unlabelled specimen from Carmira Farm

private collection; one photographic record (L. Gata,

CF).

Comment: This snake is known from several localities

in Angola (Branch 2018; Marques et al. 2018) and was

recently found in BNP (Butler et al. 2019).

Xenocalamus bicolor bicolor Gunther, 1868

Slender Quill-snouted Snake (Fig. 33A—B)

Material: Two unlabelled specimens in the private

collection of Carmira Farm.

Comment: Xenocalamus bicolor is only recorded in

Angola from a northern subspecies described in the

northeast of the country, X. b. machadoi (see Broadley

1971; Branch 2018; Marques et al. 2018). It occurs in

the Zambezi Region and adjacent western Zambia, and

it is usually associated with Kalahari sands (Branch et

Fig. 31. Psammophis mossambicus, Handa Farm (Photo by J.

Traguedo).

Amphib. Reptile Conserv.

Fig. 30. Prosymna angolensis, BNP.

al. 2019). Its presence was considered to be likely for

southeastern Angola (Branch et al. 2019), and this record

is the first for the country. The presence of supraoculars

separating the frontals from contacting the orbit (Fig.

33B), conforms to _X. b. bicolor, as does the number of

ventrals (204, 233) [Broadley 1971]. Xenocalamus b.

bicolor is known to have great intraspecific variation in

coloration, and this feature is rarely diagnostic (Broadley

1971). The patterns of both Carmira Farm specimens

resemble the “maculatus” phase described by Broadley

(1971), but more heavily marked (Fig. 33A).

Leptotyphlopidae

Namibiana aff. rostrata (Bocage, 1886)

Angolan Beaked Thread Snake (Fig. 34)

Material: NB599 (HF).

Comment: Thread snakes are a difficult group to study

due to their very small size and conservative morphology

(Broadley and Broadley 1999; Branch 2018). Namibiana

rostrata is endemic to Angola, and originally described

from Humbe, ca. 110 km south of BNP. It has recently

been recorded from BNP (Butler et al. 2019), and we

provisionally assign the specimen from Handa Farm to the

same species. Haacke (TM) collected two thread snakes,

one from Paiva Couceiro (= Quipungo, TM45190),

around 30 km southeast of Handa Farm, and another

from Hoque (TM 46699), ca. 77 km northwest of BNP,

Fig. 32. Psammophis subtaeniatus, Carmira Farm.

December 2019 | Volume 13 | Number 2 | e203

Herpetofauna of Bicuar National Park, Angola

Fig. 33. Xenocalamus bicolor bicolor, Carmira Farm. (A) Full

specimen; (B) dorsal view of head.

both identified as Leptotyohlops scutifrons scutifrons. All

these records should be carefully compared to confirm

species identification.

Pythonidae

Python anchietae Bocage, 1877

Namib Dwarf Python

Comment: Python anchietae is endemic to the Namib

Desert and adjacent areas in Angola and Namibia. It was

collected in Viriambundo (Haacke, TM), ca. 30 km west

of BNP. Although this is a historical record, it is discussed

in the species accounts here due to its relevance. This is

the fourth known record of this near-endemic species in

the country, after the type locality Catumbela (Bocage

1887), Hanha (Bogert 1940), and between Lobito and

Hanha (Laurent 1964), and the first record above the

Angolan escarpment. In Namibia, it occurs inland up to

near Windhoek, indicating that the species may extend

further inland in Angola and occur in BNP.

Amphib. Reptile Conserv.

Fig. 35. Afrotyphlops cf. schlegelii, BNP (Photo by M. Finkch).

Python natalensis Smith, 1840

Southern African Python

Material: NB794 (1); one individual observed in BNP’s

hide for game viewing (27); one photographic record

from Carmira Farm (L. Gata, CF); several photographic

records from Handa Farm (J. Traguedo, HF).

Comment: Two species of African python exist in

Angola, the northern P sebae, and the southern P

natalensis (Branch 2018). Material from BNP belongs to

P. natalensis.

Typhlopidae

Afrotyphlops cf. schlegelii (Bianconi, 1849)

Schlegel's Blind Snake (Fig. 35)

Material: Photographic record (M. Finckh, 39).

Comment: One specimen was photographed after heavy

rainfall at night (nearly 100 mm of precipitation). Dorsal

coloration was dark yellowish with scattered dark blotches

(Fig. 35) and snout-vent length measured around 70 cm

(M. Finckh, pers. comm.). Afrotyphlops schlegelii is a

sister species of A. mucruso, and both have been recorded

from Angola. Marques et al. (2018) limit A. mucruso to

northeastern Angola, and A. schlegelii to the southwest,

and Haacke (TM) also has records for Huila Province

(as Rhinotyphlops schlegelii petersii). A taxonomic

discussion is presented in Broadley and Wallach (2009),

and further discussions of both species in Angola are

presented by Conradie et al. (2016) and Branch (2018).

Confident identification requires additional material, and

the genus deserves further taxonomic revisions.

Viperidae

Bitis arietans Merrem, 1820

Puff Adder

Material: NB713 (19); one photographic record and one

specimen from Carmira Farm private collection.

Comment: Common and widespread in Angola,

recorded from several localities (Bocage 1895; Ferreira

1897; Schmidt 1933; Bogert 1940; Hellmich 1957;

Laurent 1950, 1954, 1964: Thys van der Audenaerde

1967; Manacgas 1981; Conradie et al. 2016), with few

December 2019 | Volume 13 | Number 2 | e203

Baptista et al.

Fig. 36. Causus rhombeatus, Handa Farm (Photo by J.

Traguedo).

records from the southwest (Branch 2018; Marques et al.

2018). In Angola, it is widely known as “surucucu.”

Causus rhombeatus (Lichtenstein, 1923)

Rhombic Night Adder (Fig. 36)

Material: Photographic record from Handa Farm (J.

Traguedo, HF).

Comment: The taxonomy of night adders in Angola has

long been confusing (Branch 2018). Causus bilineatus was

described from a variety of C. rhombeatus by Boulenger

(1905), but synonymies and species validity remained

problematic until a recent revision by Rasmussen

(2005), who reassessed C. bilineatus and related species,

mapped the geographic distribution of Angolan night

adders, and provided a key to the genus. Scale counts

could not be made on the specimen from Handa Farm,

although the photographed snake had a very conspicuous

V-shaped marking on the head, 25 well-separated dark

conspicuous rhombic vertebral markings occasionally

with a white contour against a paler background, and

lacked dorsolateral stripes (Fig. 36). It was therefore

assigned to C. rhombeatus, which is widespread in Angola

(Rasmussen 2005; Marques et al. 2018).

Testudines

Pelomedusidae

Pelomedusa subrufa (Bonnaterre, 1789)

Marsh Terrapin

Material: NB780 (35); photographic record (M. Finckh,

site 43).

Comment: This species is widespread in southern,

eastern, and western Africa (Branch 2012; Turtle

Taxonomy Working Group 2017). It has been recorded

from several localities in Angola (Marques et al. 2018),

and recently from BNP (Butler et al. 2019). Petzold et

al. (2014) recorded unexpected species diversity in

Amphib. Reptile Conserv.

a molecular phylogeny of Pelomedusa and referred

northern Namibian material to P. subrufa.

Testudinidae

Kinixys cf. belliana Gray, 1831

Bell’s Hingeback Tortoise

Material: NB711 (18); NB712 (17); NB509 (16) (tissue

only); photographic record (M. Finckh, site 43).

Comment: In a molecular phylogeny of Kinixys, only

K. belliana was shown to occur in Angola (Kindler et

al. 2012). However, only northern Angolan material was

included and it is very likely that K. spekii may enter

southern Angola (see Marques et al. 2018), thus the

identification as K. belliana 1s tentative. This group is

widely distributed in Angola (Marques et al. 2018) and

has recently been collected in BNP (Butler et al. 2019).

This species has been observed at several sites throughout

the park and 1s locally harvested for consumption and for

the pet trade.

Crocodilia

Crocodilydae

Crocodylus niloticus Laurenti, 1768

Nile Crocodile

Material: Interviews.

Comment: Staff from BNP referred to the existence of

this species in the Cunene River, the eastern boundary of

the park. Historical records mention its occurrence in the

area (Monard 1937b; Sim6es 1971).

Discussion

This account presents one new record for the country

(Xenocalamus bicolor bicolor), as well as records

of reptile species that are potentially new to science

(Pachydactylus, Namibiana). Butler et al. (2019)

reported eight amphibian taxa and 21 reptile taxa (two

testudines, 14 lizards, and five snakes) from BNP and

adjacent Capelongo, while this study doubles the number

of amphibian species (16), and almost quadruples

the number of snake species (18) encountered. These

increases in known species diversity are more significant

when compared to the total species counts (94, including

historical records) for the region. Despite the considerable

udpate in knowledge that this study presents, further

surveys are likely to describe additional diversity.

Closely related taxa recorded in this study and by Butler

et al. (2019), namely Lygodactylus angolensis and L.

bradfieldi, require further research and comprehensive

comparisons to confirm whether they refer to sympatric

congeneric species or different designations of the same

taxa.

Ambiguous taxonomy reflects the unresolved status

of Angolan herpetology, as the bulk of species accounts

December 2019 | Volume 13 | Number 2 | e203

Herpetofauna of Bicuar National Park, Angola

presented here refer to problematic taxonomy and a

lack of supporting studies. This is true even for the BNP

region, which is situated within the southwest, the more

thoroughly studied part of the country. This knowledge

gap reinforces the importance of promoting surveys

which include voucher specimens and DNA sample

collections published with survey results, as highlighted

previously (Russo et al. 2019). The inclusion of DNA

barcoding for identification purposes is increasingly

common (Bittencourt-Silva 2019) and provides a more

comprehensive understanding of the results. Although

not included in the scope of this publication, biopsies and

specimens collected are being used for ongoing studies

(Baptista et al., unpub. data; Lobon-Rovira et al., unpub.

data).

The herpetofauna of the BNP region is similar to that

of other regions further east where Kalahari sands are

also the dominant substrate. It shares 12 amphibian and

30 reptilian species with southeastern Angola (Conradie

et al. 2016); seven amphibian and 20 reptilian species

with western Zambia, Ngoye Falls, and surrounding

areas (Pietersen et al. 2017); and nine amphibian and five

reptilian species with recently surveyed areas of west

Zambia (Bittencourt-Silva 2019).

In a study of herpetofauna in the southern Kalahari

domain, Haacke (1984) noted the relevance ofa latitudinal

rainfall gradient as a driver of diversity in the Kalahari.

This gradient affects vegetation canopy structure on a

wide scale in Botswana (Scholes et al. 2004), and it is also

reflected in the avifauna species assemblages of BNP,

which comprise biome-restricted species from both the

northern Angolan Miombo Woodland and the southern

Zambezian Baikiaea Woodland ecoregions (Dean 2000;

BirdLife International 2018). The effect of this gradient

on the herpetofauna of BNP is not yet known, and would

be an interesting research topic for understanding the

relevance of BNP for conserving herpetofauna. Records

of species that were previously only reported from

the coastal plain of Angola, such as Python anchietae,

have shown that they occur above the escarpment. This

shows how studying the park’s herpetofauna may also

contribute to understanding the west-to-east altitudinal

and rainfall gradients, and the distribution limits of

plateau and lowland species.

Monopeltis perplexus, an endemic Angolan

amphisbaenian, was originally described from a vague

type locality: “Hanha or Capelongo” (see discussion in

Branch et al. 2018). If Capelongo (9 km northeast of

BNP) is confirmed as the type locality, M. perplexus is

likely to occur in BNP and would be sympatric with M

anchietae and M. infuscata. Sympatric distributions of

amphisbaenians highlight the importance of Kalahari

sands for fossorial species. Although fossorial species

are well represented in the BNP region, the absence

of fossorial groups such as Acontias, Typhlacontias,

and Sepsina, which have been recorded from adjacent

areas on Kalahari sands (Monard 1937b; Branch and

Amphib. Reptile Conserv.

McCartney 1992; Haacke 1997; Conradie et al. 2016;

Pietersen et al. 2017; Marques et al. 2018; Bittencourt-

Silva 2019) is likely a result of undersampling.

Rupicolous species such as Cordylus machadoi,

Matobosaurus maltzahni, Hemirhagerrhis viperina,

and Agama p. schaki were not found in the park despite

historical records, likely as a reflection of the rarity of

rocky outcrops. Substrate specificity and isolation have

important influences on the geographic distribution of

reptiles, especially lizards, and are considered better

determinants of presence than vegetation type (Bauer

and Lamb 2005; Roll et al. 2017). Endemic sand-adapted

reptiles were reported in the Kalahari (Haacke 1984).

The BNP region has a small contact zone with the larger

Kalahari sands block to the east (Missao de Pedologia

de Angola 1959; Huntley 2019). Moreover, the park is

delimited in the east by the large Cunene River, which

may be a barrier to species dispersal, much like the

Zambezi River (Pietersen et al. 2017). The combination

of factors driving speciation in the BNP region, especially

for fossorial species, is thus still unclear, and may be

clarified by assessing the genetic divergence between

BNP specimens and those from the easternmost regions

of the Kalahari sands (e.g., southeastern Angola).

Commercial and small-scale farms occupy large

portions of northwest BNP (Mendelsohn and Mendelsohn

2018). Additional disturbances include poaching and

conversion of woodlands into thickets and shrubland

following intense and frequent fires (Mendelsohn

and Mendelsohn 2018; Mendelsohn 2019). Several

mammalian species are either extinct or on the verge of

extinction in the park (Overton et al. 2017; Mendelsohn

and Mendelsohn 2018), but the consequences of these

disturbances for herpetofauna are largely unknown. Some

species are reportedly collected opportunistically for the

pet and bushmeat trades (e.g., Kinixys), a phenomenon

that has also occurred in Cangandala National Park

(Ceriaco et al. 2018c).

The establishment of protected areas generally favors

the conservation of mammals, birds, and amphibians,

while reptiles are usually neglected (Roll et al. 2017).

Likewise, the distributions of large mammals in Angola

has defined the designation of conservation areas (Huntley

et al. 2019a). BNP is an Important Bird and Biodiversity

Area, relevant for the conservation of avifauna (Dean

2000; BirdLife International 2018) that also supports

considerable reptile diversity. Some of the species likely

to occur in the park are endemic to Angola (Monopeltis

perplexus, Nucras broadleyi, Psammophylax ocellatus,

Namibiana rostrata), to Angola and Namibia (Python

anchietae), or are poorly studied (Mertensophryne

mocquardi). A better understanding of the park’s

herpetofauna should reinforce its important role in the

conservation of Angolan biodiversity.

Acknowledgements.—This research was carried out

in the framework of the Southern African Science

December 2019 | Volume 13 | Number 2 | e203

Baptista et al.

Service Centre for Climate Change and Adaptive Land

Management (SASSCAL) project, sponsored by the

German Federal Ministry of Education and Research

(BMBF) under promotion number 01LG1201M.

Collection of specimens took place under the

Memorandum of Understanding between the Angolan

Ministry of Environment (MINAMB/INBAC) and

ISCED-Huila. This work benefited from logistical and

administrative support, records of species, access to

literature, discussions on the subject, and revisions of

the manuscript. For these we thank (alphabetically):

Adam Marques, Aimy Caceres, Bicuar National Park

(Administrator José Maria Kandungo, park rangers

and staff), Brian J. Huntley, Carmira Farm (Luis Gata,

Ernesto Preto), Governo Provincial da Huila, Handa

Farm (General Joao Traguedo), ISCED-Huila (Fernanda

Lages, Filipe Rocha, Francisco Maiato, José Luis

Alexandre, Manfred Finckh, Milciades Chicomo, Paulina

Zigelski, Valter Chissingui), John Mendelsohn, Luis

Verissimo (who provided BNP boundaries and official

gazetting information), Luke Verburgt, Michael Mills,

Pedro Vaz Pinto, SAo Neto, Sasha Vasconcelos, Sidney

Novoa, and Werner Conradie. Special thanks to Alan

Channing and Wulf Haacke, who provided unpublished

records and permission to use them. NB is currently

supported by FCT contract SFRH/PD/BD/140810/2018.

Bill Branch (1946-2018) passed away during the revision

process of this paper. In the later years of his career,

Bill dedicated considerable time and effort to studying

the Angolan herpetofauna, creating local expertise, and

stimulating surveys and publications, which have led

to remarkable findings and incommensurable advances

in the knowledge of this field. It was a privilege and an

honor for the co-authors to work with Bill on this project.

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Amphib. Reptile Conserv.

Ninda Baptista is anAngolan conservation biologist with ten years of experience inthe environmental

and conservation sectors in Angola. From 2015 to 2018, Ninda worked for the Southern African

Science Service Centre for Climate Change and Adaptive Land Management (SASSCAL) Project

at Instituto Superior de Ciéncias da Educagéo da Huila (ISCED - Huila). There she conducted

herpetological surveys and monitoring, and created a herpetological collection of Angolan

specimens. Her recent experience includes working as an assistant herpetologist in the National

= Geographic/Okavango Wilderness Project, in applied conservation in Angolan scarp forests in a

Conservation Leadership Programme-funded project, and on environmental education. Ninda is an

author of scientific papers and book chapters on Angolan herpetology and ornithology, as well as

magazine articles and books for children about Angolan biodiversity.

Telmo Antonio is an Angolan biologist who recently graduated in Biology Teaching from ISCED-

Huila, with an honors thesis on the grasses of Tundavala. Telmo worked as research intern in the

SASSCAL project at ISCED-Huila from 2016 to 2018, participating in all tasks related to plants,

mammals, and herpetofauna, especially in Tundavala and Bicuar National Park. Telmo taught

high-school Biology in Lubango, Angola. He is currently a Natural Resources Management M.Sc.

student in the Namibia University of Science and Technology, within the SCIONA Project “Co-

designing conservation technologies for Iona - Skeleton Coast Transfrontier Conservation Area

(Angola - Namibia),” which is funded by the European Union.

Bill Branch (1946-2018) was born in London and worked as Curator of Herpetology at the Port

Elizabeth Museum for over 30 years (1979-2011), and upon his retirement he was appointed

as Curator Emeritus of Herpetology. His herpetological studies have focused mainly on the

systematics, phylogenetic relationships, and conservation of African reptiles. He has published

over 300 scientific papers, and numerous popular articles and books. The latter include: South

African Red Data Book of Reptiles and Amphibians (1988), Dangerous Snakes of Africa (1995,

with Steve Spawls), Field Guide to the Reptiles of Southern Africa (1998), Tortoises, Terrapins

and Turtles of Africa (2008), and Atlas and Red Data Book of the Reptiles of South Africa, Lesotho

and Swaziland (multi-authored, 2014), as well as smaller photographic guides. In 2004 Bill was

the 4" recipient of the “Exceptional Contribution to Herpetology” award of the Herpetological

Association of Africa. Bill has undertaken field work in over 16 African countries, and described

nearly 50 species, including geckos, lacertids, chameleons, cordylids, tortoises, adders, and

frogs. He supervised all tasks related to herpetology in the SASSCAL project, the creation of a

herpetofauna archive in ISCED-Huila, and many other ongoing initiatives in Angolan herpetology.

128 December 2019 | Volume 13 | Number 2 | e203

Baptista et al.

Appendix 1. List of collecting sites in Bicuar National Park, Angola, and surroundings, with coordinates (decimal degrees).

Asterisks (*) indicate sites located outside of the park. Sampling method: AS = active searching or opportunistic observation; DOR

= dead on road; T = trapping.

: : . : % ,

Site Name Latitude (°S) Longitude (°E) range tiethod Habitat type

1 BNP, road between -15.610300 14.879561 6 Nov 2017 DOR Grassland along Luconda drainage line

Tunda Gate and Main

Camp

ps BNP, near Tchiwacussi -15.189030 15.254460 6 Nov 2017 AS Secondary growth scrubland in old crop

fields, near Tambi drainage line

3 BNP -15.177040 15.228570 6 Nov 2017 AS Degraded scrubland near Tambi drainage

line

5 BNP -15.148550 14.841380 16 Mar 2018 AS Miombo woodland with considerable

bush encroachment

6 BNP -15.130452 14.683723 6 Dec 2016 AS Recently burnt open miombo woodland

7 BNP -15.129961 14.730298 6 Dec 2016 AS Temporary pond along Bicuar drainage

line

9 BNP -15.126656 14.637726 6 Dec 2016 AS Recently burnt open miombo woodland

10 BNP -15.126032 14.601210 6 Dec 2016 AS Open miombo woodland

12. BNP, road along Bicuar -15.104853 14.840320 7 Nov 2017 AS Grassland

drainage line

13. BNP -15.100510 14.845050 6 Dec 2016 AS Open miombo woodland

14. BNP, close to Main -15.096715 14.839058 2 Dec 2016 AS Leaf litter accumulated along the road, in

Camp, road to Hombo open miombo woodland

gate

15 BNP, drainage line -15.092432 14.836883 2 Dec 2016 AS Grassland

upstream Main Camp

water hole

16 BNP, road between -15.082880 14.928760 2 Dec 2016 AS Dense miombo woodland

Capelongo Gate and

Lagoa da Matemba

17 BNP, road between -15.057374 14.932907 7 Dec 2017 AS Dense miombo woodland

Capelongo Gate and

Main Camp

18 BNP, road between -15.036282 14.959543 7 Dec 2017 AS Mosaic of dense miombo woodland and

Capelongo Gate and shrubland

Main Camp

19 BNP -15.160722 14.859036 7 Dec 2017 AS Miombo woodland

20 BNP -15.129161 14.881776 2 Jul 2017 AS Open miombo woodland

21* Outskirts of BNP -14.945094 15.095214 6 Nov 2017 DOR Degraded open shrubland

25 Lagoa da Matemba -15.122320 14.902980 4 Nov 2017 AS Permanent water body with aquatic

vegetation

26 Lagoa do Dyjimbi -15.145520 14.914670 5 Nov 2017 AS Permanent water body

27 Main Camp water whole -15.102406 14.836857 8 Dec 2016 AS Permanent water body

28 Main Camp water whole -15.098320 14.837030 7 Dec 2016 AS Permanent water body

29 Main Camp water whole -15.098060 14.836990 7 Dec 2016 AS Permanent water body

30 ‘Drainage line upstream -15.093490 14.837450 4 Dec 2016 AS Grassland

Main Camp water whole

31 BNP Main Camp -15.100725 14.839557 2009-2017 AS Human facilities, sandy soils, tall Burkea

africana trees, surrounded miombo

woodland

32* Outskirts of BNP -15.060080 15.251800 6 Nov 2017 AS Degraded open shrubland

33. BNP -15.150060 14.812300 5 Nov 2017 AS Miombo woodland with considerable

bush encroachment

34 BNP, near Bicuar drain- -15.243460 14.891460 5 Nov 2017 AS Recently burnt open miombo woodland

age line

35 BNP, road between -15.024986 14.796841 4 Nov 2017 AS Open miombo woodland with regenerat-

Hombo Gate and Main ing understorey

Camp

Amphib. Reptile Conserv. 129 December 2019 | Volume 13 | Number 2 | e203

Date or date

Sampling

Herpetofauna of Bicuar National Park, Angola

Appendix 1. List of collecting sites in Bicuar National Park, Angola, and surroundings, with coordinates (decimal degrees).

Asterisks (*) indicate sites located outside of the park. Sampling method: AS = active searching or opportunistic observation; DOR

= dead on road; T = trapping.

Date ordate Sampling

‘ ‘ A ‘ 2% ‘

Site Name Latitude (°S) Longitude (°E) ranige method Habitat type

36 ~~ BNP, close to Main -15.095409 14.838751 2 Nov 2016 AS Miombo woodland

Camp, road to Hombo

gate

39 BNP, Tunda Gate -15.645000 14.708333 6 Dec 2015 AS Dry woodland on deep sands

40 BNP -15.133300 14.900000 11 Dec 2015 AS Ecotone between grassland and wood-

land

41* Lagoa Nougalafa, out- -14.977117 14.693638 6 Dec 2016 AS Permanent water body

skirts of BNP

42* Between Chibemba and -16.03 14.20 15 Feb 2010 AS Small wetland, approximate coordinates

Cahama

43. BNP -15.042928 14.805010 5 Dec 2016 AS Border of one a grassland valley, in the

ecotone between suffrutex grasslands

and woodland, crossing sandy dirt road

CF* Carmira Farm -16.044722 14.569167 2015-2018 AS Baikiaea/Burkea woodland, deep Kala-

hari sands

C1* Road to Carmira Farm -15.992383 14.410287 5 Apr 2018 AS Baikiaea/Burkea woodland, deep Kala-

hari sands

HF* Handa Farm -14.699722 14.295833 2015-2018 AS Degraded miombo woodland with over-

grazing and deforestation

ast Woodland trapline 1 -15.094405 14.838312 2-7 Dec T Miombo woodland not burnt for more

2016 than 1 year

T2 Woodland trapline 2 -15.100358 14.838958 7-10 Dec Ak Miombo woodland not burnt for more

2016 than 5 years

T3 Grassland trapline -15.102190 14.837057 2-10 Dec Mt Grassland near permanent body of water

2016

Amphib. Reptile Conserv. 130 December 2019 | Volume 13 | Number 2 | e203

Official journal website:

amphibian-reptile-conservation.org

Amphibian & Reptile Conservation

13(2) [General Section]: 1-13 (e183).

First record of the Cat Ba Tiger Gecko, Goniurosaurus

catbaensis, from Ha Long Bay, Quang Ninh Province,

Vietnam: microhabitat selection, potential distribution,

and evidence of threats

19.10Hai Ngoc Ngo, 7Tuan Quang Le, *Minh Le Pham, 2?Truong Quang Nguyen,

56.7Minh Duc Le, Mona van Schingen, and *:"°*Thomas Ziegler

'Vietnam National Museum of Nature, Vietnam Academy of Science and Technology, 18 Hoang Quoc Viet Road, Hanoi, VIETNAM Institute of

Ecology and Biological Resources, Vietnam Academy of Science and Technology, 18 Hoang Quoc Viet Road, Hanoi, VIETNAM°?Graduate University

of Science and Technology, Vietnam Academy of Science and Technology, 18 Hoang Quoc Viet Road, Hanoi, VIETNAM *Ha Long Bay Management

Department, 166 Le Thanh Tong Road, Ha Long, Ha Long City, Quang Ninh, VIETNAM *Faculty of Environmental Sciences, Hanoi University of

Science, Vietnam National University, 334 Nguyen Trai Road, Hanoi, VIETNAM °Central Institute for Natural Resources and Environmental Studies,

Hanoi National University, 19 Le Thanh Tong, Hanoi, VIETNAM ‘Department of Herpetology, American Museum of Natural History, Central Park

West at 79 Street, New York, New York 10024, USA *Federal Agency for Nature Conservation, CITIES Scientific Authority, Konstantinstrasse 110,

53179 Bonn, GERMANY * Institute of Zoology, University of Cologne, Ziilpicher StraBe 47b, 50674, KéIn, GERMANY '°Cologne Zoo, Riehler Strape

173, 50735, KélIn, GERMANY

Abstract.—The Cat Ba Tiger Gecko (Goniurosaurus catbaensis) was described from Cat Ba Island, Hai Phong,

northern Vietnam in 2008, while a presumed congener was recently spotted from another offshore island in

the Ha Long Bay. During the field surveys reported here, new Goniurosaurus occurrences were discovered

for the first time on small offshore islands in the Ha Long Bay, Quang Ninh Province. These were identified

and confirmed as G. catbaensis based on morphological and molecular data. However, these newly found

populations are very small and exposed to increasing anthropogenic pressures. Since knowledge about the

species ecology remains poor, the first microhabitat characterization for G. catbaensis is provided herein, which

is essential for conservation of the species as well as its natural habitats. Sex- and age-related differences in

selection of perch height are herein presented. In addition, we present evidence for various anthropogenic

threats such as regular trade in living tiger geckos (including G. catbaensis) on local markets in Hai Phong and

Ho Chi Minh cities, Vietnam. These findings highlight the need for more stringent conservation measures to

reduce human impacts on the extremely small, insular populations of the Cat Ba Tiger Gecko.

Key words. Anthropogenic pressure, conservation, ecology, offshore islands, phylogram, trade

Citation: Ngo HN, Le TQ, Pham ML, Nguyen TQ, Le MD, van Schingen M, Ziegler T. 2019. First record of the Cat Ba Tiger Gecko, Goniurosaurus

catbaensis, from Ha Long Bay, Quang Ninh Province, Vietnam: microhabitat selection, potential distribution, and evidence of threats. Amphibian &

Reptile Conservation 13(2) [General Section]: 1-13 (e183).

International (CC BY 4.0): https://creativecommons.org/licenses/by/4.0/], which permits unrestricted use, distribution, and reproduction in any me-

dium, provided the original author and source are credited. The official and authorized publication credit sources, which will be duly enforced, are as

follows: official journal title Amphibian & Reptile Conservation; official journal website: amphibian-reptile-conservation.org.

Submitted: 18 July 2017; Accepted: 27 February 2019; Published: 2 September 2019

Introduction araneus, G. catbaensis, G. huuliensis, G. lichtenfelderi,

and G. /uii (Nguyen et al. 2009). Among these species, the

The genus Goniurosaurus currently comprises 19

species with a disjunct distribution in southern East Asia.

Most Goniurosaurus species are endemic with restricted

distribution ranges (Chen et al. 2014; Grismer et al. 1994,

1999; Honda and Ota 2017; Seufer et al. 2005; Yang and

Chan 2015; Zhou et al. 2018; Ziegler et al. 2008). Habitat

degradation and overharvesting for the pet trade were

identified as major threats to wild populations of tiger

geckos (Yang and Chan 2015). At present, five species

of Goniurosaurus are known from Vietnam, namely G.

Correspondence. * ziegler@koelnerzoo.de

Amphib. Reptile Conserv.

insular Cat Ba Tiger Gecko (Goniurosaurus catbaensis)

was discovered on Cat Ba Island in Cat Hai District,

Hai Phong City, northeastern Vietnam, where it was

assumed to be endemic (Ziegler et al. 2008). Preliminary

population assessments of G. catbaensis revealed that its

effective population size, defined as number of mature

individuals, is much smaller than the suggested threshold

values for minimal viable populations to maintain a stable

population in the long term (Ngo et al. 2016; Nguyen

et al. 2016, 2018; Reed et al. 2003; Traill et al. 2007).

September 2019 | Volume 13 | Number 2 | e183

Goniurosaurus catbaensis in Ha Long Bay, Vietnam

i op ae Ae , ; at! t ea rah

Fig. 1. New population. (A) Habitat of Goniurosaurus

be? ee # te a OP EAs)

catbaensis on one offshore island in

+' wide’ ‘ie of : * :

Laer “S ee ae =.

Ro Se ef Po

Ha Long Bay, Quang Ninh Province:

(B) Microhabitat of G. catbaensis in Ha Long Bay; (C) Adult male; and (D) Adult female from Ha Long Bay. Photos: H.N. Ngo.

Even in undisturbed habitats, G. catbaensis occurs at low

densities (Ngo et al. 2016; Nguyen et al. 2016, 2018).

The insular Cat Ba Tiger Gecko was found to be

vulnerable to anthropogenic disturbances, and of high

demand in pet markets as well as on Internet platforms

(Ngo et al. 2016; Nguyen et al. 2018). In addition to

poaching, habitat destruction for touristic purposes

has dramatically increased the pressure on the wild

G. catbaensis population. Consequently, the need for

protection of the Cat Ba Tiger Gecko has received

growing attention. Based on the first international

population and trade investigations, this species has

recently been listed in the IUCN Red List of Threatened

Species as "Endangered" (Nguyen et al. 2016). The

wild population is probably in peril due to its restricted

distribution range, rising anthropogenic threats, and the

lack of appropriate conservation measures. For the latter,

detailed information on habitat requirements and the

exact distribution of this species is essential, but such

data are currently lacking. Ngo et al. (2016) recently

suggested the potential occurrence of G. catbaensis on at

least one more offshore island in Ha Long Bay.

To confirm this possibility, we investigated other small

offshore islands in Ha Long Bay, Quang Ninh Province

to locate populations of G. catbaensis, and predicted the

overall availability of suitable habitats for the species in

northeast Vietnam. In addition, the present study aimed

to provide the first data on microhabitat selection of G.

catbaensis. We assumed that differences in habitat use

would occur between age classes and sexes, as they have

Amphib. Reptile Conserv.

been observed in other lizards (Snyder et al. 2010; van

Schingen et al. 2015).

Materials and Methods

Study areas: Study sites were selected based on our

previous surveys on Cat Ba Island, Hai Phong City,

and on photo documentation which gave evidence for

the possible occurrence of Goniurosaurus on a small

island in Ha Long Bay, Quang Ninh Province (Ngo et

al. 2016). Cat Ba Island and adjacent islands comprise

isolated limestone karst formations, which provide

diverse habitats for a unique flora and fauna (Clements

et al. 2006). Cat Ba Archipelago was recognized as the

“Cat Ba Archipelago Biosphere Reserve” (CBBR) by

the United Nations Educational, Scientific and Cultural

Organization (UNESCO) in 2004 due to its significant

ecosystem and biodiversity values (CBBR Authority

2013). Ha Long Bay was also twice recognized (in 1994

and 2000) by UNESCO as a World Heritage Site for the

outstanding universal value of its landscape, geology, and

geomorphology (The Management Department of Ha

Long Bay 2014). Both areas are among the most popular

tourist destinations in Vietnam, and face challenges from

rapid tourism development.

Field surveys: Field surveys were conducted on Cat

Ba Island between June and August 2014, May 2015,

and during a short time in June 2016, which fell in the

non-hibernation season of Goniurosaurus (Grismer et

September 2019 | Volume 13 | Number 2 | e183

Ngo et al.

al. 1999; Ngo et al. 2016). Furthermore, six offshore

islands in Ha Long Bay, situated in close proximity to

Cat Ba Archipelago, were surveyed in July 2016. Night

excursions were conducted between 7:30 and 11:30

PM, when the lizards were found to be active (Ngo et

al. 2016; Ziegler et al. 2008). To measure morphological

characters, the animals were captured by hand and

subsequently released at the same spot after checking and

taking measurements.

Ecological analyses: Microhabitat data were recorded

for each sighted G. catbaensis, including substrate

types (classified as cliff, rock, branch, sand, or forest

floor), perch height (vertical distance between captured

animal and ground, in cm), percentage of vegetation or

cave coverage, position (resting outside or inside cave),

substrate surface condition (dry or wet), and activity

(resting, feeding, or foraging). Air temperature and

relative humidity were measured with a digital thermo-

hygrometer (TFA Dostmann/Wertheim Kat. Nr. 30.5015),

and substrate temperature and body surface temperature

of animals were measured with an infrared thermometer

(Measupro IRT20).

To identify intraspecific differences in microhabitat

selection by G. catbaensis, individuals were classified

into different age classes according to their snout-vent

lengths (SVL): SVL < 85 mm = juvenile, SVL > 85 mm

and < 105 mm = sub-adult, and SVL => 105 mm = adult

(Ngo et al. 2016). Adults were differentiated between

the sexes, as well as between gravid and non-gravid

individuals. Sex of specimens was determined by the

presence of the large swollen hemipenal bulges in males,

while non-swollen in females.

A t-test, with @ = 0.05, was performed to determine

differences in microhabitat parameters between age

classes and sexes. Statistical analyses were performed

with the program PAST, Version 2.17c (Hammer et al.

2001).

Morphological analyses: Morphometric measurements

of captured individuals were taken with a digital caliper

to the nearest 0.1 mm. In addition, two voucher specimens

of the newly discovered populations in Ha Long Bay were

collected, euthanized with ethylacetate, preserved in 70%

ethanol, and deposited in the collections of the Vietnam

National Museum of Nature (VNMN), Hanoi, Vietnam

(VNMN 05423, VNMN 05424). Morphological characters

were taken according to Ngo et al. (2016), Orlov et al.

(2008), Yang and Chan (2015), and Ziegler et al. (2008).

Abbreviations of measurements are as follows: snout

vent length (SVL) from tip of snout to vent; tail length

(TaL) from vent to tip of tail; distance between axilla

and groin (AG) from posterior edge of forelimb insertion

to anterior edge of hind limb insertion; forelimb length

(FoL) from axilla to tip of longest finger; hindlimb length

(HiL) from groin to tip of longest finger; snout to eye

distance (SE) from tip of snout to anterior-most point of

eye; eye to ear distance (EE) from posterior margin of eye

to posterior margin of ear; orbital diameter (OD) greatest

diameter of orbit; ear diameter (ED) longest dimension

of ear; internarial distance (IND) as distance between

nares; anterior eye distance (AED) as distance between

Amphib. Reptile Conserv.

anterior corners of eyelids; posterior eye distance

(PED) as distance between posterior corners of eyelids;

maximum head width (HW); maximum head height

(HH); head length (HL) from tip of snout to posterior

edge of occiput; pileus length (PL) from tip of snout to

posterior scale of the head; and jaw length (JL).

Abbreviations of scalation are as follows: supralabials

(SPL); infralabials (IFL); nasal scales surrounding

nare (N); internasals (IN); gular scales bordering the

internasals (PostIN); postmentals (PM); gular scales

bordering the postmentals (GP); eyelid fringe scales

or ciliaria (CIL); granular scales surrounding dorsal

tubercles (GST); dorsal tubercle rows at midbody

(DTR); paravertebral tubercles between limb insertions

(TL); scales around midbody (MB); subdigital lamellae

under the first finger (LF1) and the fourth finger (LF4),;

subdigital lamellae under the first toe (LT1) and the

fourth toe (LT4); precloacal pores (PP); and postcloacal

tubercles (PAT).

Molecular analyses: To confirm the taxonomic status of

the newly collected Goniurosaurus from Ha Long Bay,

Quang Ninh Province, a fragment of the mitochondrial

16S ribosomal gene was amplified, using the primer pair

16Sar and 16Sbr (Palumbi et al. 1991), for three samples

(VNMN 05424 plus two small tissue samples from

two released individuals, field numbers G8 and G12).

Tissue samples were taken from the tail tips, which were

disinfected before immediate release of the animals at the

site of capture. DNA was extracted from tissue samples

using the DNeasy blood and tissue kit, Qiagen (Redwood

City, CA). The extracted DNA from the fresh tissue

samples were amplified by PCR, with the PCR volume

(21ul) consisting of 10 ul of mastermix (Fermentas,

Canada), 5 ul of water, 2 ul of each primer at 10 pmol/

ul, and 2 wl of DNA. The PCR conditions were: 95 °C

for five minutes to activate the taq; with 40 cycles at 95

°C for 30s, 50 °C for 45s, 72 °C for 60s; and the final

extension at 72 °C for six minutes (Ngo et al. 2016).

PCR products were subjected to electrophoresis

through a 1% agarose gel (UltraPure™, Invitrogen). Gels

were stained for 10 minutes in 1x TBE buffer at 2 pg/

ml of ethidium-bromide, and visualized under UV light.

Successful amplifications were purified to eliminate

PCR components using GeneJET™ PCR Purifcation Kit

(Fermentas, Canada). Purified PCR products were sent to

lst Base (Selangor, Malaysia) for sequencing. Sequences

were edited using the program Geneious v.7.1.8 (Kearse et

al. 2012). After sequences were aligned using Clustal X v2

(Thompson et al. 1997), data were analyzed by Bayesian

inference as implemented in MrBayes v3.2 (Ronquist

et al. 2012). Settings for these analyses followed Le et

al. (2006), except that the number of generations in the

Bayesian analysis was increased to 1x10’. The optimal

model for nucleotide evolution was set to GTR+I+G as

selected by Modeltest v3.7 (Posada and Crandall 1998).

The cutoff point for the burn-in function was set to 13 in

the Bayesian analysis, as -InL scores reached stationarity

after 13,000 generations in both runs. Nodal support was

evaluated using posterior probability in MrBayes v3.2.

Uncorrected pairwise divergences were calculated in

PAUP*4.0b10 (Swofford 2001).

September 2019 | Volume 13 | Number 2 | e183

Goniurosaurus catbaensis in Ha Long Bay, Vietnam

Gekko gecko (AB028758)

Goniurosaurus kuroiwae (AB028766)

99 Goniurosaurus yingdeensis (KC900231)

Goniurosaurus zhelongi (KJ423105)

-Goniurosaurus araneus (AB308460)

Goniurosaurus luli (EU499390)

Goniurosaurus Iuii ML19 (MK041068)

73 Goniurosaurus luii TDLS2012.1 (MKO41069)

100

gut ee neunts luli (KC765083)

Goniurosaurus luli (KC 765084)

99 Goniurosaurus luli TAQ182 (MK041067)

92 26

Goniurosaurus luli IEBR3254 (MK041070)

Goniurosaurus liboensis (KC900230)

Goniurosaurus catbaensis (EU499389)

Cat Ba Island

él 100 || Goniurosaurus catbaensis VNMN05424 (MK041071)

Goniurosaurus catbaensis G8 (MK041072)

Other Islands

in Ha Long Bay

Goniurosaurus catbaensis G12 (MK041073)

100 Goniurosaurus lichtenfelderi (JF799756)

Goniurosaurus hainanensis (KC765080)

0.05

Fig. 2. Phylogram of Goniurosaurus based on the Bayesian analysis of a 16S ribosomal fragment. Numbers next to nodes are

Bayesian posterior probabilities. Voucher numbers of new samples and GenBank accession numbers are placed after species names

and in parentheses, respectively.

Species distribution models (SDMs): Based on

occurrence records and a set of 19 environmental factors,

the current overall availability of suitable habitats for G.

catbaensis were predicted using the program Maxent

Version 3.3.3.e (Beaumont et al. 2005; Phillips et al.

2006). Only the most distant occurrences of each site

were included in the analyses to minimize effects of

spatial autocorrelation and to ensure the independence

of the records (Jennings and Veron 2011; Jennings et al.

2013). As a result, 11 records were filtered from a total

of 60 localities of G. catbaensis on Cat Ba Island and

Ha Long Bay. Nineteen bioclimatic variables that were

obtained from the WorldClim global climate database

(http://www.worldclim.org, accessed September 2016;

Hijmans et al. 2005; Table 1) were used as environmental

predictors.

Threat records: To get a first impression of trade in

Goniurosaurus species in Vietnam, local pet markets

were visited in Hai Phong and Ho Chi Minh cities, the

two most important trade centers in the country, and

different Internet platforms were investigated. Two local

dealers from Ho Chi Minh City offering Goniurosaurus

online were interviewed in September 2016, in order

to trace the source of the traded Goniurosaurus species

Amphib. Reptile Conserv.

in Vietnam. Additionally, five fishermen from the Ha

Long Bay were interviewed to identify caves used by

tourism companies for night parties, and determine

the general attitude and use of the species in Ha Long

Bay. Those sites located within the World Heritage Site

were subsequently surveyed in July 2016 to evaluate

potential threats from tourism activities. The names of

interviewees are kept anonymous to ensure data privacy

rights and Internet links are not disclosed to prevent

misuse. Accurate locality data, cave names, and prices

are also not presented to prevent targeted poaching for

the wildlife trade.

Results

New records of Goniurosaurus catbaensis: During

the present study, new Goniurosaurus occurrences were

discovered on four small offshore islands, including two

tourism caves in Ha Long Bay, Quang Ninh Province.

The distances between these islands ranged from 1.4 km

to 13 km, while the shortest distance between Cat Ba

Island and one surveyed island in Ha Long Bay was 1.2

km. A total of 14 individuals (eight males, four females,

one juvenile, and one unsexed individual which was only

photographed) were recorded on these islands, which

September 2019 | Volume 13 | Number 2 | e183

Ngo et al.

Table 1. Bioclimatic variables from environmental data (Source: http://www.worldclim.org, accessed September 2016).

No. Bioclimatic variables from the WorldClim dataset

l BIO1 = “Annual Mean Temperature”

2 BIO2 = “Mean Diurnal Range" (Mean of monthly [max temp - min temp])

3 BIO3 = "Isothermality" (P2/P7) (*100)

4 BIO4 = "Temperature Seasonality" (standard deviation * 100)

5 BIO5 = "Max Temperature of Warmest Month"

6 BIO6 ="Min Temperature of Coldest Month"

7 BIO7 ="Temperature Annual Range" (P5—P6)

8 BIO8 = "Mean Temperature of Wettest Quarter"

9 BIO9 = "Mean Temperature of Driest Quarter"

10 BIO10 = "Mean Temperature of Warmest Quarter"

1] BIO11 ="Mean Temperature of Coldest Quarter"

12 BIO12 = "Annual Precipitation (year)"

13 BIO13 = "Precipitation of Wettest Month"

14 BIO14 = "Precipitation of Driest Month"

15 BIO15 = "Precipitation Seasonality" (Coefficient of Variation)

16 BIO16 = "Precipitation of Wettest Quarter"

17 BIO17 = "Precipitation of Driest Quarter"

18 BIO18 = "Precipitation of Warmest Quarter"

19 BIO19 = "Precipitation of Coldest Quarter"

ranged between 0.34 and 2.94 km? in size.

Molecular analysis using Bayesian inference of

the obtained matrix containing 613 aligned characters

showed that all samples from Cat Ba Island (n = 1) and

from the most distant other islands in Ha Long Bay (n=3)

clustered in a single clade with strong statistical support

(posterior probability = 100%, Fig. 2). Genetic analyses

revealed that sequences of the new records from Ha

Long Bay, Quang Ninh Province, were identical to each

other and virtually the same (99% to 100%) as that of the

holotype of G. catbaensis from Cat Ba Island (GenBank

accession number: EU499389). The maximum genetic

divergence between the samples is approximately 0.3%,

whereas the lowest divergence between two species of

this genus, i.e., G. hainanensis and G. lichtenfelderi, is

approximately 2.3% (Table 2). These results confirmed

the newly recorded Goniurosaurus populations in Ha

Long Bay are conspecific with G. catbaensis from Cat

Ba Island (Fig. 2).

In addition, the morphological characters of the newly

recorded G. catbaensis from Ha Long Bay accorded well

with the population from Cat Ba Island, except that three

of six individuals from a single site in Ha Long Bay

showed a postrostral (internasal) scale. This character 1s

consistently lacking 1n individuals recorded so far from

Cat Ba Island (Ziegler et al. 2008) [Fig. 3A, 3B; Table 3].

Microhabitat selection: A total of 61 sightings took

place (13 from smaller islands in the Ha Long Bay,

and 48 from Cat Ba Island). Goniurosaurus catbaensis

was active in the surroundings of large limestone caves

covered in part by primary forest vegetation and in the

vicinity of primary shrub vegetation on limestone. Mean

air temperatures were 28.1 + 1.7 °C (21.5-31.3 °C, n=

Long Bay. Photos H.N. Ngo.

Amphib. Reptile Conserv.

Fig. 3. Absence versus occasional presence of internasal scales of Goniurosaurus catbaensis from (A) Cat Ba Island and (B) Ha

September 2019 | Volume 13 | Number 2 | e183

Goniurosaurus catbaensis in Ha Long Bay, Vietnam

Forest

floor 5%

i=)

‘)

hel

Fig. 4. (A) Substrate selection of Goniurosaurus catbaensis. (B) Box plots of perch heights of different age classes and sexes.

59) slightly higher than mean substrate temperatures of

26.02 + 1.5 °C (22.2—28.2 °C, n= 28, Table 4). Recorded

relative humidity at microsites ranged between 70-99%

(mean 84.9 + 6.99%, n = 52).

A vast majority of lizards was found on limestone

cliffs (62%), followed by rocks (28%), while only a few

specimens were found on the forest floor (5%), branches

(3%), or sand (2%) [Fig. 4A]. A significantly lower

number of lizards was encountered inside compared

to outside of limestone caves (26.9% vs. 73.1%,

respectively). Goniurosaurus catbaensis selected spots

with a mean canopy coverage of 95.2 + 9.6% (n = 63,

Table 4). Adult specimens (non-gravid) were found at

average heights of 1.15 m (n = 38), while juveniles and

gravid females resided at significantly lower heights of

0.28 m(n=4) and 0.41 m (n= 12), respectively (t= 2.82,

df = 48, P < 0.05; t = 2.06, df = 40, P < 0.05, Fig. 4B).

A majority (about 77.4%, n = 48) of lizards was resting

during the surveys, while only a few individuals (n = 14)

were found actively foraging.

Suitable habitats for G. catbaensis were predicted to

encompass a majority of small islands belonging to Cat

Ba Island and Ha Long Bay, and include a wider area on

the coastal mainland of Quang Ninh Province, where no

surveys have been conducted so far (Fig. 5).

Trade: Trade in living tiger geckos has been frequently

recorded by our team in local pet markets from Hai

Phong and Ho Chi Minh cities, as well as on Facebook

since 2015. Interviews with two local traders in Ho Chi

Minh City revealed that they pay for local villagers living

within the species’ distribution range to collect live tiger

geckos during the non-hibernation season, confirming

the wild (rather than captive-bred) source of traded

animals. The dealers reportedly received individuals of

three tiger gecko species, namely G. huuliensis, G. luii,

and G. catbaensis, collected in April 2015. Among those,

three individuals of G. huuliensis (one male and two

females) were allegedly collected by a local hunter from

Huu Lien Nature Reserve, Lang Son Province. Two local

collectors from Cao Bang Province reportedly collected

six individuals (three males and three females) of G. /uii

in northern Vietnam and another local hunter collected

two couples of G. catbaensis. These 13 wild caught tiger

geckos were transferred to pet markets in Ho Chi Minh

City, southern Vietnam, in April 2015.

Human impacts on the habitat: Tourism activities in

the region have dramatically increased in the past, and

likely exerted enormous pressure on wild G. catbaensis

populations. Events organized by tourism companies

Table 2. Uncorrected (“p”) distance matrix showing percentage pairwise genetic divergence (16S) between members of

Goniurosaurus.

Species name 1 2 3 4 5 6 7 8 9

1. G. araneus —

2. G. catbaensis 6.4-6.7 -

3. G. hainanensis [3x 12.4-12.8 -

4. G. kuroiwae 20.4 19.5-19.8 1.3 —

5. G. liboensis 6.3 6.6-6.8 12.8 21.9 —

6. G. lichtenfelderi 12.9 11.2-11.6 23 18.8 13:3 —

7. G. luii 5.6-6.2 6.2-7.1 12.2-12.9 20.0-204 3.43.8 11.5-13.5 —

8. G. yingdeensis 14.8 13.4-13.5 15.2 18.8 13.0 Las2 13.3-13.5 —

9. G. zhelongi 15.3 14.2-14.4 16.9 21.4 13.4 16.2 14.8-15.4 48 —

Amphib. Reptile Conserv. 6 September 2019 | Volume 13 | Number 2 | e183

Ngo et al.

106°30'0"E 107°O'0"E

107°30'0"E 108°0'0"E

21°30'0"N

21°0'0"N

21°30'0"N

21°0'0"N

Legend

===== National boundary

Province boundary

Habitat suitability

——— Low

20°30'0"N

106°30'0"E 107°0'0"E

0 15 30

20°30'0"N

107°30'0"E

60 Kilometers

108°0'0"E

Fig. 5. Predicted habitat suitability for Goniurosaurus catbaensis in Vietnam.

regularly took place in at least two caves located

within the UNESCO World Heritage Site. According to

interviews with fishermen, daily excursions to the caves

are scheduled to start at 7:30 PM and end around 11:00

PM. On these occasions, tourists dine in brightly lit

caves before returning to their tour boats (Fig. 6B). As

a consequence, wildlife is likely to be disturbed by the

candle light, noisy sounds, and waste left by the tourists.

Discussion

New population records: Since its discovery in 2008,

the Cat Ba Tiger Gecko was thought to be endemic to

Cat Ba Island (Ziegler et al. 2008). These new records

of G. cf. catbaensis on further offshore islands in Ha

Long Bay confirmed for the first time the occurrence of

the species outside its type locality. The newly recorded

specimens showed an insignificant genetic divergence

from the type series from Cat Ba Island and could be

assigned to G. catbaensis (Table 2). Accordingly, the

newly collected specimens from Ha Long Bay were also

almost identical to the type series of G. catbaensis in

morphology, except for the presence of a single internasal

scale (which 1s absent in the type series from Cat Ba, see

Ziegler et al. 2008) in a few individuals from a single

site in Ha Long Bay. These findings indicated a slightly

broader distribution range of the species than previously

expected.

According to Li et al. (2010), the islands of Ha Long

Bay and Cat Ba Archipelago were shaped by the erosion

of limestone karst formations within the Gulf of Tonkin

Amphib. Reptile Conserv.

at the northern east coast of Vietnam after the coastal

shelf region became inundated by marine waters about

13,000 years ago. Repeated falls (> 50 m) of the sea

level during glaciations periodically connected various

islands and the mainland, which allowed exchanges

between island and mainland populations, as well as

colonization and re-colonization between island and

mainland populations (Li et al. 2010; Liang et al. 2018).

Thus, past recurrent gene flow is assumed to have

occurred between (sub)populations, which helped to

maintain a classical island-mainland metapopulation—in

accordance with the high genetic similarity between G.

catbaensis (sub)populations from different islands with

identical habitats (Hanski 1991; Harrison and Taylor

1997; Levins 1969). Orlov et al. (2008) confirmed

that G. lichtenfelderi was found from both continental

mainland and islands. On the other hand, Liang et al.

(2018) suggested that G. lichtenfelderi diverged from

G. hainanensis of Hainan Island to Vietnam (including

both mainland and island populations), which might have

occurred during the glacial periods with past dispersal

events. The speciation in the diversification process of

Goniurosaurus was probably promoted by the adaption to

different microhabitats. Populations of G. lichtenfelderi

were found on granite beds of valley streams, while the

closely related G. hainaensis is found on igneous rocks

and G. catbaensis occurs in karst forests (Orlov et al.

2008; Liang et al. 2018; Ziegler et al. 2008; Nguyen et

al. 2018).

To avoid the misuse of distribution data for targeted

harvesting of the species (e.g., Lindenmayer and Scheele

September 2019 | Volume 13 | Number 2 | e183

Goniurosaurus catbaensis in Ha Long Bay, Vietnam

Table 3. Morphological characters of Goniurosaurus from Ha Long Bay, Quang Ninh Province, compared with G. catbaensis from

Cat Ba Island, Hai Phong Province. Measurements are given in mm. Note: (*) n= 6; (*)n=2.

Specimens

SVL

TaL

PostIN

CIL

GST

DTR

LF1

LF4

LT1

LT4

PAT

2017; Stuart et al. 2006; Yang and Chan 2015), detailed

locality information of the new records is being withheld.

According to the SDMs G. catbaensis is predicted to

occur on other, similar islands in the Gulf of Tonkin, but

Ha Long Bay (current study, n = 13)

74.54-122.7 (111.2 + 11.9)

10.1-97.6 (69.9 +27)

33.9-60.2 (52.9+6.5)

21.3-33.8 (30.2 +2.9)

14.4-24.56 (22.1 + 2.5)

7.1-14.9 (12.8 + 1.9)

32.2-53.8 (50.4+ 5.6)

42-67.47 (60.1 + 6.2)

8.7-13.4 (11.9 + 1.1)

9.4-13.2 (10.8 + 1.5)*

5.6-8.3 (7.5 +0.7)

2.8-5.3 (4.01 + 0.8)

3.39-4.33 (3.9 + 0.34)*

6.78-8.62 (7.98 + 0.67)*

11.8-15.03 (13.9 + 1.23)*

12.3-10.8 (18.1 +2.1)

27.6-32.5 (29.9 + 1.7)*

3.5-3.8 (3.68 + 0.1)

1.9-2.3 (2.140.1)

1.28-1.48 (1.37 + 0.05)

2.3-3.01 (2.35 + 1.6)

0.9-1.2 (1.1 £0.1)*

9-10 (9.4 +0.5)*

8-9 (9.75 + 0.45)*

5-6 (5.25 + 0.5)

0-1 (0.23 + 0.4)

0-2 (0.4 + 0.77)

2-3 (2.5+0.7)8

45-49 (46.75 + 1.7)"

104-109 (106.5 + 3.5)"

9-12 (10.5+1.3)8

35-37 (36 + 1.4)"

234

9-12 (10.25 + 1.3)"

18-19 (18.75 + 0.5)"

9-10 (9.75 + 0.5)

240

20-24 (22.5 +1.4)*

1-3 (2.25 + 0.6)*

Amphib. Reptile Conserv.

Cat Ba Island (current study, n = 48)

69.2-130.4 (108.9 + 12.6)

28.9-104.02 (78 + 17.7)

43.07-58.43 (48.44 5.4)*

17.8-34.2 (29.8 + 3.5)

13.9-28.2 (21.9 +2.5)

8.2-16.9(12.4+ 1.9)

29.7-54 (47.8 + 4.7)

36.2-65 (57.9 + 5,99)

10.45-13.4 (12.1 + 1.0)*

9.78-12.13 (11.1 + 0.88)*

6.1-8.95 (7.6 + 1.1)*

2.8-4.3 (3.540.5)*

2.9-4.2 (3.74 0.5)*

6.9-8.47 (7.5 + 0.6)*

11.9-15.1 (13.1 +1.3)*

15.5-19.5 (17.2 + 1.4)*

26.6-32.8 (29.2 +2.5)*

3.3-4.3 (3.74 0.2)

2.04-2.45 (2.3 +0.14)*

1.1-1.5 (1.36 + 0.09)

1.79-3.3 (2.4+ 0.4)

1.07-1.1 (1.09 + 1.14)*

8-11 (10.08 + 1.1)*

8-10 (8.8 +0.7)*

6-8 (7+ 0.47)*

2-3 (2.83 + 0.41)*

6-9 (7.8+1.2)*

41-56 (47.8 + 4.4)*

102-109 (103.8 + 3.8)*

9-14 (10.3 + 1.6)*

27-34 (31.5 + 3.0)*

19-25 (22.3 + 1.97)*

9-11 (10 +0.7)*

19-20 (19.3 + 0.5)*

9-10 (9.91 +0.3)*

22-24 (23.4 + 0.8)*

21*

2-3 (2.5+40.5)*

Cat Ba Island (Ziegler et al. 2008) [n = 4]

84.7-111.5 (102.4 + 14.5)

§2.5-101.5 (68.1 + 27.6)

23.1-30.6 (27.7 +4.1)

16.2-21.6 (19.5 +2.9)

10.1-14.3 (12.2 + 2.0)

9.8-12.6 (11.5 + 1.6)

8.5-12.3 (10.6 +2.1)

3.61-3.67 (3.7 + 0.05)

1.43-2.11 (1.6 +0.4)

2.29-2.43 (2.33 + 0.07)

1.02-1.15 (1.09 + 0.07)

8-9 (8.7 + 0.5)

6-8 (7.8 + 0.6)

5-6 (5.1 40.4)

2-3 (2.8 + 0.5)

6-7 (7.22 + 0.6)

52-55 (54.0+ 1.1)

112-127 (119.2 + 7.6)

8-11 (9.8 + 1.6)

33-34 (33.7 + 0.6)

23-25 (24.0 + 1.2)

11-12 (11.75 + 0.5)

18-19 (18.1 +£0.5)

11-12 (11.44 0.6)

22-24 (23.440.7)

5-21 (15.3 +2.5)

2-3 (2.8 + 0.5)

is still endemic to Ha Long Bay and Cat Ba Archipelago.

According to Orlov et al. (2008) the type locality of

G. lichtenfelderi is an offshore island in Bai Tu Long

Archipelago, which is contiguous with Ha Long Bay

September 2019 | Volume 13 | Number 2 | e183

Ngo et al.

—see i _

Fig. 6. Potential threats to Goniurosaurus. (A) Flooding of Viet Hai Commune in August 2015. (B) Tourist event in a cave within

the UNESCO World Heritage Site on Ha Long Bay. Photos H.N. Ngo.

in the Gulf of Tonkin. However, extensive field surveys

have failed to record any individual of G. catbaensis,

occurring in syntopy with G. lichtenfelderi (Gawor et al.

2016; Nguyen et al. 2011; Orlov et al. 2008). The habitat

of G. lichtenfelderi in Bai Tu Long was described as

valleys of forest streams on granite rocks within mixed

forests of bamboo and broad-leaved trees (Gawor et al.

2016; Nguyen et al. 2009; Nguyen et al. 2011; Orlov et

al. 2008; Ziegler et al. 2008), while G. catbaensis was

found only in limestone karst ecosystems present in Ha

Long and Cat Ba archipelagos. Accordingly, our SDMs

predicted the potential distribution of G. catbaensis to

encompass Ha Long Bay and Cat Ba Archipelago, but

excluding Bai Tu Long Archipelago (Fig. 5). However,

the present SDMs also predicted the mainland area

including limestone formations around Ha Long City to

be suitable for G. catbaensis. Thus, it will be important to

search for further occurrences at these predicted sites in

order to determine the exact distribution boundaries, and

to assess genetic diversity of potentially new populations.

Microhabitat selection: Both sex- and age-related

perch selection were found in G. catbaensis, namely

differences in perch heights. Specifically, juveniles and

gravid females occurred at significantly lower heights

than subadults and adults. Similar habitat divergences

between juveniles and adult individuals have been

reported for Crocodile Lizards in Vietnam (van Schingen

et al. 2015), and gekkonids in New Caledonia (Snyder et

al. 2010).

This study also revealed that the body surface

temperature of G. catbaensis showed a highly positive

correlation with the air temperature (7, = 0.56; P < 0.05,

n = 23) and substrate temperature (7, = 0.66; P < 0.001,

n= 26). Thus, as in other ectotherms, basic physiological

functions of G. catbaensis, such as locomotion, growth,

and reproduction are determined by the environmental

temperature. Since tropical lizards are considered to

have narrow temperature optima, and only few options

for behavioral and physiological compensation, they

are assumed to be especially vulnerable to extinction by

climate warming (Deutsch et al. 2008; Doody and Moore

2010; Huey et al. 2009; Vié et al. 2009). In particular,

body surface temperatures of G. catbaensis ranged from

between 23.6 and 30.6 °C (mean = 27.2 + 1.6 °C, n= 26)

and were comparably higher than those of G. kuroiwae

with average skin surface temperatures of 16.6 °C in the

humid subtropical Oriental forest (Werner et al. 2005).

Potential threats and recommendations for

conservation: Due to the restricted distribution range,

low densities, and estimated global population being

much lower than suggested threshold values for minimal

viable populations, the Cat Ba Tiger Gecko is expected

to be especially endangered to unsustainable for harvest

(Ngo et al. 2016). Consequently, the species was recently

assessed and ranked by the IUCN Red List of Threatened

Species as "Endangered" (Nguyen et al. 2016). Other

members of the genus Goniurosaurus from Vietnam

have not been considered for inclusion on the IUCN

Red List yet, as data on their population statuses are

currently lacking. The findings reported here indicate

that not only G. catbaensis, but also G. huuliensis and

G. luii, are subject to intensive collection for local trade

and provide concrete evidence for the wild source of

the respective specimens. It is likely that the reported

Table 4. Environmental parameters characterizing the microhabitat selection of Goniurosaurus catbaensis.

Parameter Number of sightings (n)

Canopy cover [%] 63

Height [m] 54

Elevation [m asl] 60

Air Temperature [°C] 59

Substrate Temperature [°C] 28

Relative air Humidity [%] 52

Amphib. Reptile Conserv.

Min Max Mean + SD

50 100 95.2+9.6

0 3 0.97 + 0.86

4 132 46.2 + 32.9

21-8 313 28.1+1.7

222 28.2 26.02 + 1.5

70 99 84.9 + 6.99

September 2019 | Volume 13 | Number 2 | e183

Goniurosaurus catbaensis in Ha Long Bay, Vietnam

cases only reflect a small proportion of illegal harvesting

activities. Since over-exploitation of local populations

of range-restricted lizard species has been repeatedly

found to rapidly cause extinction (e.g., Aultya et al.

2016; Stuart et al. 2006; Yang and Chang 2015), further

research on the population status, distribution, ecology,

and availability of suitable microhabitat sites is critically

needed. The results of such studies may lead to the

elevation or determination of the conservation status of

other tiger gecko species and provide critical scientific

data for future captive breeding programs. To reduce

poaching and to control the trade in wild Goniurosaurus,

we recommend continued monitoring of the scales and

patterns of trade in combination with aforementioned

population assessments. We also strongly advise

against providing exact locality information for new

Goniurosaurus populations in future publications, as this

action might increase poaching activities at respective

sites (Lindenmayer and Scheele 2017; Stuart et al. 2006;

Yang and Chan 2015).

In addition to the illegal collection of animals, human

impacts on habitats have dramatically increased by

means of expanding tourism activities (see also Ngo et

al. 2016). Tourism events in caves, causing disturbance

by candle light, noisy sounds, and waste might result in

the extirpation of G. catbaensis within these limestone

caves. We suggest that tourism companies should hold

such events only on their boats to reduce disturbances

in the cave habitats of G. catbaensis, or at least restrict

tourist access to only limited, selected islands.

Following Ngo et al. (2016), the sites in Viet Hai

Village on Cat Ba Island had been recommended as a

priority conservation zone for species conservation,

since G. catbaensis was found to be most abundant at

those sites. However, during the most recent survey in

July 2016, no specimens of G. catbaensis were observed

in Viet Hai Commune. We assume that an extensive flood

in August 2015 might have killed a large amount of the

local wildlife, including the Cat Ba Tiger Gecko, at this

site. Viet Hai Commune was isolated for a week after

torrential rains brought the water level up to the roofs of

local houses. Since G. catbaensis was found to generally

occur at low elevation ranges (4-132 m asl), and Viet

Hai is situated only up to 36 m asl (see Fig. 6B), this

species is particularly vulnerable to natural catastrophes

such as storms, floods, and sea level rises, throughout

its distribution range (see Dessler 2016; Saunders et al.

1991). Since local populations are extremely small, they

are especially prone to extinction by catastrophic events.

The devastating consequences of such natural disasters

underline the importance of maintaining numerous

independent subpopulations in order to compensate for

such events.

In summary, the insular (sub)populations of G.

catbaensis are threatened by harvest for the pet trade,

human activities within its habitats, and natural

catastrophes such as increasingly extreme floods and

storms in northeastern Vietnam, probably triggered

by climate change (The Governmental Committee on

Flood and Storm Prevention 2016). Thus, we herewith

emphasize the importance of setting aside priority

conservation zones for this species, in order to establish

Amphib. Reptile Conserv.

a connected and buffered system that allows (sub)

populations to recover from catastrophes. We also

recommend the establishment of an assurance population,

i.e., an ex situ conservation breeding program for the

species. Although such an effort has been started at the

Me Linh Station for Biodiversity (see Ziegler et al. 2016)

in Northern Vietnam, more resources need to be allocated

to enhance the effort to conserve the species.

Acknowledgements.—For supporting field work and

issuing relevant permits, we thank the authorities of the

Cat Ba National Park (CBNP), Hai Phong City, and the

Management Department of Ha Long Bay (MDHLB),

Quang Ninh Province. We are very thankful to L. Barthel

(University of Cologne) and K.X. Nguyen (CBNP) for

assistance in the field. We are grateful to T. Pagel and C.

Landsberg (Cologne Zoo); M. Bonkowski (University of

Cologne); C.X. Le, T.H. Tran, T.H. Vu, C.T. Pham, and

T.V. Nguyen (IEBR, Hanoi); M.T. Nguyen, L.V. Vu, and

T.T. Nguyen (VNMN, Hanoi); and V.Q. Luu (VNUF, Ha

Noi) for their support of conservation-based biodiversity

research in Vietnam. Thanks to H.T. Ngo for laboratory

assistance. Our research was funded by Cologne Zoo, the

Mohamed bin Zayed Species Conservation fund (Project:

170515492), the National Foundation for Science and

Technology Development (NAFOSTED, Grant No. 106-

NN.06-2016.59), the Idea Wild, and Vietnam National

Museum of Nature (VNMN). Cologne Zoo is a partner of

the World Association of Zoos and Aquariums (WAZA):

Conservation Project 07011 (Herpetodiversity Research).

Research of Hai Ngoc Ngo in Germany is funded by the

German Academic Exchange Service (DAAD).

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September 2019 | Volume 13 | Number 2 | e183

Ngo et al.

Hai Ngo Ngoc is a Ph.D. candidate at the Institute of Zoology, University of Cologne, and the Cologne

Zoo, Germany. He has worked as a researcher at the Vietnam National Museum of Nature since 2014,

and finished his M.Sc. degree in 2015 from the University of Science, Vietnam National University,

Hanoi. Hai has participated in diverse herpetological surveys in Vietnam and has experience in field

research and conservation work. His focus is on the ecology, phylogeny, and conservation of endemic

and endangered reptile species in Vietnam.

Tuan Quang Le is a researcher at the Institute of Ecology and Biological Resources (IEBR), Vietnam

Academy of Science and Technology (VAST). He is conducting Ph.D. research at Academia Sinica,

Taiwan, and majored in ecology, ecological modeling, and remote sensing. His current research

focuses on the application of remote sensing and geographical information systems in the ecology

and ecological modeling of mammals and reptiles. In particular, Tuan is using species distribution

modeling to predict the potential distributions of species, including reptiles and mammals, in Vietnam.

In addition, he evaluates the impacts of climate change on those species based on future climate

scenarios, and is also developing remote sensing-based approaches for mapping species habitats

Minh Le Pham is a researcher at the Ha Long Bay Management Department. He finished his M.Sc.

degree in 2018 at the Vietnam National University, Hanoi. His work focuses on the ecology and

conservation of biodiversity and landscapes in the Ha Long Bay World Heritage Site.

Truong Quang Nguyen is a researcher at the Institute of Ecology and Biological Resources (IEBR),

Vietnam Academy of Science and Technology (VAST), and is a member of the Biodiversity and Nature

Conservation projects of the Cologne Zoo. Truong finished his Ph.D. in 2011 at the Zoological Research

Museum Alexander Koenig (ZFMK) and the University of Bonn, Germany, as a DAAD Fellow. From

2011 to 2014, he worked as a postdoctoral student in the Zoological Institute, University of Cologne.

Truong has conducted numerous field surveys and is the co-author of 12 books and more than 300

papers related to biodiversity research and conservation in Southeast Asia. His research interests focus

on the systematics, ecology, and phylogeny of reptiles and amphibians from Southeast Asia.

Minh Duc Le has been working on conservation-related issues in Southeast Asia for more than 15

years. His work focuses on biotic surveys, wildlife trade, and conservation genetics of various wildlife

groups in Indochina. Minh is currently working on projects which characterize genetic diversity

of highly threatened reptiles and mammals in the region, and he has pioneered the application of

molecular tools in surveying critically endangered species in Vietnam. He has long been involved in

studying the impact of the wildlife trade on biodiversity conservation in Vietnam, and is developing a

multidisciplinary framework to address this issue in the country.

Mona van Schingen finished her Ph.D. on the Vietnamese Crocodile Lizard in 2017 at the Institute

of Zoology of the University of Cologne and the Cologne Zoo, Germany. Since 2011, Mona has been

investigating the herpetofauna of Vietnam, in the working group of Thomas Ziegler. She has conducted

diverse field excursions to Vietnam and is engaged in several research, conservation, and awareness

projects focusing on various species in Vietnam. Since 2017 she has been working for the German

CITIES Scientific Authority at the Federal Agency for Nature Conservation.

Thomas Ziegler has been the Curator of the Aquarium/Terrarium Department of the Cologne Zoo

since 2003, and is the coordinator of the Cologne Zoo’s Biodiversity Research and Nature Conservation

Projects in Vietnam and Laos. Thomas studied biology at the University Bonn (Germany), and

conducted his diploma and doctoral thesis at the Zoological Research Museum Alexander Koenig

in Bonn, with a focus on zoological systematics and amphibian and reptile diversity. Thomas has

been engaged with herpetological diversity research and conservation in Vietnam since 1997. As a

zoo curator and project coordinator, he tries to combine in situ and ex situ approaches—viz., to link

zoo biological aspects with diversity research and conservation in the Cologne Zoo, as well as in

rescue stations and breeding facilities in Vietnam and in Indochina’s last remaining forests. Thomas is

a professor at the Institute of Zoology of Cologne University. Since 1994, he has published more than

430 papers and books, mainly dealing with herpetological diversity.

Amphib. Reptile Conserv. 13 September 2019 | Volume 13 | Number 2 | e183

Official journal website:

amphibian-reptile-conservation.org

Amphibian & Reptile Conservation

13(2) [General Section]: 14—27 (e187).

urn:lsid:zoobank.org:pub:7F259D5F-3CBF-4421-BA54-5CECDB687340

A new species of dwarf day gecko (Reptilia: Gekkonidae:

Cnemaspis) from lower-elevations of Samanala Nature

Reserve in Central massif, Sri Lanka

1*Suranjan Karunarathna and 7Kanishka D.B. Ukuwela

'Nature Explorations and Education Team, No: B-1 / G-6, De Soysapura Flats, Moratuwa 10400, SRI LANKA *Department of Biological Sciences,

Faculty of Applied Sciences, Rajarata University of Sri Lanka, Mihintale 50300, SRI LANKA

Abstract.—A new day gecko species of genus Cnemaspis Strauch, 1887 is described from a midland forested

area of Udamaliboda (north-western foothills of Samanala Nature Reserve) in Sri Lanka. This species is

medium in size (30-35 mm SVL) and can be differentiated from all other Sri Lankan congeners by a suite

of distinct morphometric, meristic, and color characters (dorsum with smooth and homogeneous granular

scales; chin, gular, pectoral, and abdominal scales smooth; precloacal pores absent in males, 14—15 femoral

pores separated by 9-11 unpored interfemoral scales in males; subcaudals smooth, subhexagonal, enlarged,

subequal, forming a regular median row). It was recorded from tall trees with smooth bark in home gardens,

and also on clay walls in very old tall houses in wet, cool, and shady forests, distributed across mid elevations

(~450-—650 m) with limited anthropogenic disturbance. They can climb to heights of 7 m on vertical surfaces of

trees. The most noteworthy behavior of this species is that when “scared,” it runs only upward to the canopy

of the tree or along the wall to hide within crevices. The major threats for this species in Udamaliboda and other

locations in lower Samanala Nature Reserve are habitat loss due to expansion of commercial-scale agriculture

and monoculture plantations, and illicit forest encroachments. Therefore, these foothill forests warrant special

conservation, habitat protection, further in-depth research, and specific hands-on management practices.

Keywords. Arboreal, conservation, ecology, rainforest, redlist, taxonomy, Sripadha, threats

Citation: Karunarathna S, Ukuwela KDB. 2019. A new species of dwarf day gecko (Reptilia: Gekkonidae: Cnemaspis) from lower-elevations of

Samanala Nature Reserve in Central massif, Sri Lanka. Amphibian & Reptile Conservation 13(2) [General Section]: 14-27 (e187).

License [Attribution 4.0 International (CC BY 4.0): https://creativecommons.org/licenses/by/4.0/], which permits unrestricted use, distribution, and

reproduction in any medium, provided the original author and source are credited. The official and authorized publication credit sources, which will

be duly enforced, are as follows: official journal title Amphibian & Reptile Conservation; official journal website: amphibian-reptile-conservation.org.

Received: 16 May 2019; Accepted: 29 July 2019; Published: 9 September 2019.

Introduction

Sri Lanka’s wet zone, home to unique assemblages of

floral and faunal communities with high endemism, is

one of the smallest biodiversity hotspots in the world

(Meegaskumbura et al. 2002; Gunawardene et al. 2007).

Within the diverse reptile community of the island (~225

species), the diversity of geckos (family Gekkonidae) 1s

remarkable; 54 species (in eight genera) have been de-

scribed so far, accounting for 24% of the overall reptilian

species richness (Somaweera and Somaweera 2009; de

Silva and Ukuwela 2017). Of these, 44 species (~81%)

are endemic and 45 species (~83%) are threatened (MoE-

SL 2012, Karunarathna et al. 2019a,b). The genus Cne-

maspis comprises 32 species in Sri Lanka and all of them

are endemic (Batuwita et al. 2019; Karunarathna et al.

2019a,b; de Silva et al. 2019). Sri Lankan Cnemaspis

Correspondence. * suranjan.karu@gmail.com

Amphib. Reptile Conserv.

Species represent two distinct evolutionary lineages,

the C. kandiana and C. podihuna clades (Agarwal et al.

2017; Karunarathna et al. 2019b). The high species rich-

ness in Sri Lanka may be due to multiple possible colo-

nization events from the Indian mainland with isolated in

situ speciation (Agarwal et al. 2017).

During the past decade, the number of species recog-

nized in the genus Cnemaspis globally has grown rap-

idly, reaching over 150 species (Grismer et al. 2014; Uetz

et al. 2019; Karunarathna et al. 2019b), and Cnemaspis

is now the second most speciose gecko genus in the Old

World (Rosler et al. 2019). Though Sri Lanka is a small

island, it harbors about 21% of the world’s Cnemaspis

species, of which 90% have been described in the last two

decades, including many described only recently (Ka-

runarathna et al. 2019b). However, most of the Cnemas-

pis species from the dry and intermediate climatic zones

September 2019 | Volume 13 | Number 2 | e187

Karunarathna and Ukuwela

of Sri Lanka are restricted to small isolated hillocks scat-

tered in the lowlands (Karunarathna et al. 2019b). Future

studies on the biogeography of Cnemaspis in Sri Lanka

are expected to highlight the importance of these isolated

habitats in generating and maintaining the diversity of

these unique groups of geckos on the island. During a

field excursion to Udamaliboda (northwest Samanala

Nature Reserve), an unidentified Cnemaspis species was

discovered that closely resembles C. gemunu, C. godage-

darai, C. phillipsi, and C. scalpensis, and it is described

here as a new species.

Materials and Methods

Specimen collection. Museum acronyms follow Sabaj

Pérez (2015). The type material discussed in this pa-

per is deposited in the National Museum of Sri Lanka

(NMSL), Colombo. Specimens were hand-caught and

photographed in life. Three specimens were euthanized

using halothane, fixed in 10% formaldehyde for two

days, washed in water, and transferred to 70% ethanol

for long-term storage. Tail tips were collected (as tissue

samples) before fixation in formaldehyde for future ge-

netic analyses and stored in 95% ethanol under relatively

cool conditions (20-25 °C). For comparison, we exam-

ined 402 Cnemaspis specimens (catalogued and uncata-

logued) representing all recognized Sri Lankan species,

including all type specimens housed at the NMSL and

The Natural History Museum, London (BMNH). Speci-

mens that formerly belonged to the Wildlife Heritage

Trust (WHT) collection and bear WHT numbers are cur-

rently deposited in the NMSL, catalogued under their

original numbers. Original specimens in this study were

collected during a survey on lizards of Sri Lanka under

permit numbers WL/3/2/1/14/12 and WL/3/2/42/18 (A

and B), issued by the Department of Wildlife Conserva-

tion, and permit numbers FRC/5 and FRC/6, issued by

the Forest Department of Sri Lanka. Additional informa-

tion on morphology and natural history of Sri Lankan

Cnemaspis species was extracted from the relevant lit-

erature (Bauer et al. 2007; Manamendra-Arachchi et

al. 2007; Wickramasinghe and Munindradasa 2007; Vi-

danapathirana et al. 2014; Amarasinghe and Campbell

2016; Wickramasinghe et al. 2016; Batuwita and Udu-

gampala 2017; Agarwal et al. 2017; Batuwita et al. 2019;

Karunarathna et al. 2019b). Assignment of unidentified

specimens to species was based on the presence of shared

morphometric and meristic characters (Wickramasinghe

et al. 2016; Batuwita and Udugampala 2017; Agarwal et

al. 2017; Batuwita et al. 2019; Karunarathna et al. 2019b;

de Silva et al. 2019).

Morphometric characters. Forty morphometric mea-

surements were taken (to the nearest 0.1 mm) using a Mi-

tutoyo digital Vernier calliper, and detailed observations

of scales and other structures were made through Leica

Wild M3Z and Leica EZ4 dissecting microscopes. The

Amphib. Reptile Conserv.

following symmetrical morphometric characters were

taken on the left side of the body: eye diameter (ED),

horizontal diameter of eye ball; orbital diameter (OD),

greatest diameter of orbit; eye to nostril length (EN), dis-

tance between anteriormost point of orbit and posterior

border of nostril; snout length (ES), distance between an-

teriormost point of orbit and tip of snout; snout to nostril

length (SN), distance between tip of snout and anterior-

most point of nostril; nostril width (NW), maximum hori-

zontal width of nostrils; eye to ear distance (EE), distance

between posterior border of eye and anteriormost point

of ear opening; snout to axilla distance (SA), distance be-

tween axilla and tip of snout; ear length (EL), maximum

length of ear opening; interorbital width (IO), shortest

distance between left and right supraciliary scale rows;

inter-ear distance (IE) distance across head between the

two ear openings; head length (HL), distance between

posterior edge of mandible and tip of snout; head width

(HW), maximum width of head between the ears and the

orbits; head depth (HD), maximum height of head at lev-

el of eye; jaw length (JL), distance between tip of snout

and corner of mouth; internarial distance (IN), smallest

distance between inner margins of nostrils; snout to ear

distance (SED), distance between tip of snout and ante-

riormost point of ear; upper-arm length (UAL), distance

between axilla and angle of elbow; lower-arm length

(LAL), distance from elbow to wrist with palm flexed;

palm length (PAL), distance between wrist (carpus) and

tip of longest finger excluding the claw; length of digits

I-V of manus (DLM), distance between juncture of the

basal phalanx with the adjacent digit and the tip of the

digit, excluding the claw; snout-vent length (SVL), dis-

tance between tip of snout and anterior margin of vent:

trunk length (TRL), distance between axilla and groin;

trunk width (TW), maximum width of body; trunk depth

(TD), maximum depth of body; femur length (FEL), dis-

tance between groin and knee; tibia length (TBL), dis-

tance from knee to ankle with heel flexed; heel length

(HEL), distance between ankle (tarsus) and tip of longest

toe (excluding the claw) with both foot and tibia flexed;

length of pedal digits I-V (DLP), distance between junc-

ture of basal phalanx with the adjacent digit and the digit

tip, excluding the claw; tail length (TAL), distance be-

tween anterior margin of the vent and tail tip; tail base

depth (TBD), maximum height of tail base; and tail base

width (TBW), widest point of tail base.

Meristic characters. Twenty-nine discrete characters

were observed and recorded using Leica Wild M3Z and

Leica EZ4 dissecting microscopes on both left and right

sides of the body (reported in the form L/R): numbers of

supralabials (SUP) and infralabials (INF), between first

labial scale and corner of the mouth; number of inter-

orbital scales (INOS), between left and right supracili-

ary scale rows; number of postmentals (PM) bounded by

chin scales, 1‘ infralabial on left and right and the men-

tal; number of chin scales (CHS), scales touching medial

September 2019 | Volume 13 | Number 2 | e187

New species of Cnemaspis from Sri Lanka

— 600m

— 1200m

Arid Zone

| Dry zone

Bi Intermediate zone

| Wet zone

| | Montane Forest

Kilometers

WGS 1984 UTM Zone 44N

Fig. 1. Currently known distribution of Cnemaspis anslemi

sp. nov. (Udamaliboda-star), and related species: C. phillipsi

(Gammaduwa-traingle), C. scalpensis (Gannoruwa—square),

C. gemunu (Haggala-—circle), and C. godagedarai (Ensalwatte—

diamond) in Sri Lanka.

edge of infralabials and mental between juncture of 1*

and 2™ infralabials on left and right; number of suprana-

sal (SUN), scales between nares; presence of postnasal

(PON), scales posterior to naris; presence of internasal

(INT), scale between supranasals; number of supraciliary

scales (SUS), above eye; number of scales between eye

and tympanum (BET), from posteriormost point of orbit

to anteriormost point of tympanum; number of canthal

scales (CAS), number of scales from posteriormost point

of naris to anteriormost point of the orbit; total lamel-

lae on manus I-V (SLM), counted from first proximal

enlarged scansor greater than twice width of largest palm

scale, to distalmost lamella at tip of digits; number of

dorsal paravertebral granules (PG), between pelvic and

pectoral limb insertion points along a straight line im-

mediately left of vertebral column; number of midbody

scales (MBS), from center of mid-dorsal row diagonal-

ly toward ventral scales; number of midventral scales

(MVS), from first scale posterior to mental to last scale

anterior to vent; number of belly scales (BLS), across ven-

ter between lowest rows of granular dorsal scales; total

Amphib. Reptile Conserv.

lamellae on pes I-V (SLP), counted from first proximal

enlarged scansor greater than twice the width of largest

heel scale, to distalmost lamella at tip of digits; number

of femoral pores (FP), present on femur; number of non-

pored posterior femoral scales (PFS), counted from distal

end of femoral pore row to knee; and interfemoral scales

(IFS), number of non-pored scales between first femoral

pores on both femurs. In addition, the texture (smooth

or keeled) of ventral scales, the texture (homogeneous

or heterogeneous) of dorsal scales, the number of spi-

nous scales on flanks (FLSP), and characteristics such as

appearance of caudal scales (except in specimens with

regenerated tails) were also recorded. Coloration was

determined from digital images of living specimens and

also from direct observations tn the field.

Natural history. The new species described here was

collected on a field survey conducted in Udamaliboda,

Samanala Nature Reserve of Sri Lanka (Fig. 1). Be-

havioral and other aspects of the natural history of the

focal species were observed through opportunistic and

non-systematic means. Such observations were made

at a minimum distance of 2-4 m from the focal animals

while taking precautions to avoid disturbances. To record

elevation and georeference species locations, an eTrex®

20 GPS (Garmin) was used. Sex was determined by the

presence (male) or absence (female) of femoral pores.

The conservation status of the species was evaluated us-

ing the 2001 IUCN Red List Categories and Criteria ver-

sion 3.1 (UCN 2012).

Systematics

Cnemaspis anslemi sp. nov.

urn:Isid:zoobank.org:act: A5B58BCF-0BEE-4714-971 B-B38218F74956

Anslems’ Day Gecko (English)

Anslemge divaseri hoona (Sinhala)

Anslemvin pahalpalli (Tamil)

Figs. 2-5; Tables 1-2

Holotype. NMSL.2019.14.01, adult male, 34.4 mm SVL

(Fig. 2), collected from a tall, straight tree with good

canopy cover in a home garden (bordering forest) in

Udamaliboda, Kegalle District, Sabaragamuwa Province,

Sri Lanka (6.859728°N, 80.448736°E, WGS1984;

elevation 485 m, around 16.00 hrs) on 25 March 2019 by

Suranjan Karunarathna and Kanishka Ukuwela.

Paratypes. NMSL.2019.14.02, adult female, 32.5 mm

SVL collected from an old clay house wall (bordering

forest) in Udamaliboda, Kegalle District, Sabaragamuwa

Province, Sri Lanka (6.869611°N, 80.457069°E,

WGS1984; elevation 634 m, around 10.00 hrs) on 26

March 2019 by Suranjan Karunarathna and Kanishka

Ukuwela, and NMSL.2019.14.03, adult female, 30.0 mm

SVL (Fig. 3) collected from a tall, straight tree with good

canopy cover in a home garden (bordering the forest) in

September 2019 | Volume 13 | Number 2 | e187

Karunarathna and Ukuwela

Fig. 2. Close-ups of Cnemaspis anslemi sp. nov. male holotype

weil »

(NMSL.2019.14.01): (A) dors

f ‘ C wh

> . A

f x

al, (B) lateral, (C) ventral aspects of

head, (D) scales on lateral surface of trunk, (E) smooth ventral scales, (F) homogeneous dorsal scales, (G) cloacal characters with

femoral pores, (H) subdigital lamellae on pes, (I) subdigital lamellae on manus, (J) lateral side of tail, (IK) oval shaped subcaudals,

and (L) dorsal scalation of tail. Photos: Suranjan Karunarathna.

Udamaliboda, Kegalle District, Sabaragamuwa Province,

Sri Lanka (6.859728°N, 80.448736°E, WGS1984;

elevation 485 m, around 14.00 hrs), on 27 March 2019

by Suranjan Karunarathna and Kanishka Ukuwela.

Diagnosis. Cnemaspis anslemi sp. nov. can be readily

distinguished from its Sri Lankan congeners by a

combination of the following morphological and meristic

Amphib. Reptile Conserv.

characteristics, and also color pattern: maximum SVL

34.4 mm; dorsum with homogeneous, smooth granular

scales; 2/2 supranasals, one internasal, and 1/1 postnasal

present; three enlarged postmentals; postmentals

bounded by five chin scales; chin and gular scales

smooth, granular, juxtaposed; pectoral and abdominal

scales smooth and subimbricate; 3—5 well developed

tubercles on posterior flank; 118-122 paravertebral

September 2019 | Volume 13 | Number 2 | e187

New species of Cnemaspis from Sri Lanka

Fig. 3. Dorsal and ventral aspects of the type series of Cnemaspis anslemi sp. nov. (A) Male holotype, NMSL.2019.14.01, (B) fe-

male paratype, NMSL.2019.14.02, and (C) female paratype, NMSL.2019.14.03 from Udamaliboda, Samanala Nature Reserve, Sri

Lanka. Photos: Suranjan Karunarathna.

granules linearly arranged; 19-21 belly scales across

venter; precloacal pores absent in males, 14—15 femoral

pores on each side in males separated by 9-11 unpored

interfemoral scales in males, and 2—3 unpored posterior

femoral scales in males; 111-117 ventral scales; 87-91

midbody scales; subcaudals smooth, subhexagonal,

enlarged, subequal, forming a regular median row; 8—9

supralabials; 8—9 infralabials; 16-17 total lamellae on

digit IV of manus, and 20—21 total lamellae on digit I'V of

pes (Table 1). Dorsal body reticulated brown, black, and

white; two large oval patches present on the neck; chin

and gular with bright yellow, and femur dirty yellow.

Comparisons with other species. Based on the

presence of enlarged hexagonal subcaudal scales C.

anslemi sp. nov. can be assigned to the C. podihuna

clade sensu Agarwal et al. (2017). However, the new

species may be readily differentiated from congeners in

this clade as follows: from C. kandambyi Batuwita and

Udugampala, 2017, C. molligodai Wickramasinghe and

Munindradasa, 2007, and C. podihuna Deraniyagala,

1944 by absence (versus presence) of precloacal pores;

from C. alwisi Wickramasinghe and Munindradasa,

2007, C. godagedarai de Silva et al. 2019, C. hitihami

Karunarathna et al. 2019, C. kohukumburai Karunarathna

et al. 2019, C. phillipsi Manamendra-Arachchi et al.

2007, C. punctata Manamendra-Arachchi et al. 2007,

C. rajakarunai Wickramasinghe et al. 2016, and C.

rammalensis Vidanapathirana et al. 2014 by the presence

of fewer ventral scales (111-117 versus 145-153, 133-

137, 132-135, 131-134, 128-143, 129-137, 146-186,

and 186—207, respectively); from C. ni/gala Karunarathna

et al. 2019 by the presence of more femoral pores (14—

15 versus 7-9), from C. gemunu Bauer et al. 2007 by

the presence of a greater number of belly scales (19-21

versus 13-16) and by presence of more paravertebral

granules (118—122 versus 79-93); and from C. scalpensis

Amphib. Reptile Conserv.

(Ferguson, 1877) by the presence of fewer tubercles on

posterior flank (3—5 versus 9-11) and a greater number of

paravertebral granules (118-122 versus 102-112).

Among species of the C. kandiana clade sensu

Agarwal et al. (2017), C. anslemi sp. nov. differs by the

absence (versus presence) of precloacal pores and the

presence (versus absence) of clearly enlarged, hexagonal,

or subhexagonal subcaudal scales from the following

species: C. amith Manamendra-Arachchi et al. 2007,

C. butewai Karunarathna et al. 2019, C. gotaimbarai

Karunarathna et al. 2019, C. ingerorum Batuwita et al.

2019, C. kallima Manamendra-Arachchi et al. 2007, C.

kandiana (Kelaart, 1852), C. kivulegedarai Karunarathna

et al. 2019, C. kumarasinghei Wickramasinghe and

Munindradasa, 2007, C. latha Manamendra-Arachchi

et al. 2007, C. menikay Manamendra-Arachchi et al.

2007, C. nandimithrai Karunarathna et al. 2019, C.

pava Manamendra-Arachchi et al. 2007, C. pulchra

Manamendra-Arachchi et al. 2007, C. retigalensis

Wickramasinghe and Munindradasa, 2007, C.

samanalensis Wickramasinghe and Munindradasa,

2007, C. silvula Manamendra-Arachchi et al. 2007,

C. tropidogaster (Boulenger, 1885) and C. upendrai

Manamendra-Arachchi et al. 2007.

Description of Holotype. An adult male, 34.4 mm SVL.

Body slender and relatively long (TRL 42.3% of SVL).

Head relatively large (HL 30.3% of SVL, HL 71.6% of

TRL), narrow (HW 17.2% of SVL, HW 56.7% of HL),

depressed (HD 10.0% of SVL, HD 33.1% of HL) and

distinct from neck. Snout relatively long (ES 80.7% of

HW, ES 45.8% of HL), more than twice the eye diameter

(ED 38.4% of ES), more than half the length of jaw (ES

67.8% of JL), snout slightly concave in lateral view; eye

relatively small (ED 17.6% of HL), twice as large as ear

(EL 34.4% of ED), pupil rounded; orbit length greater

than eye to ear distance (OD 115.6% of EE) and greater

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Karunarathna and Ukuwela

Table 1. Morphometric and meristic data of holotype and paratypes of Cnemaspis anslemi sp. nov. from Udamaliboda, Sri Lanka.

Abbreviations: Holo—holotype, Para—paratype, M—male, F—female, L—left, R-right.

NMSL NMSL NMSL NMSL NMSL NMSL

Measurement 2019.14.01 2019.14.02 2019.14.03 Counts 2019.14.01 2019.14.02 2019.14.03

Holo (M) Para (F) Para (F) Holo (M) Para (F) Para (F)

SVL 34.4 32,3 30.3 FLSP (L/R) 5/5 3/3 4/3

ED 1.8 1.8 1.8 SUP (L/R) 8/8 9/8 8/8

OD 3.3 39 3.1 INF (L/R) 9/8 8/8 8/8

EN 2.9 oo ih INOS 27 ZY 26

ES 4.8 4.7 4.6 PM 3 5 3

SN 1,3 bal 1.1 CHS 5 3 ES

NW ORF OZ 0.2 SUN (L/R) 2/2 2/2 2/2

EE 29 De Df PON (L/R) 1/1 1/1 1/1

SA 16.9 LS 15<2 INT 1 1 1

EL 0.6 0.6 0.6 SUS (L/R) 9/10 11/11 10/9

IO 3.4 aS 3:3 BET (L/R) 18/18 18/17 19/18

IE 4.8 4.7 4.7 CAS (L/R) 9/10 8/8 9/8

HL 10.4 99 a9 TLM (i) (L/R) 11/11 10/11 10/10

HW 59 5.8 ae TLM (ii) (L/R) 12/12 12/13 12/11

HD 3.5 Br 3.0 TLM (iit) (L/R) 14/13 14/14 13/13

JL 7.0 6.9 6.9 TLM (iv) (L/R) 17/17 17/16 17/17

IN 1.7 1.8 1.7 TLM (v) (L/R) 13/13 13/13 13/12

SED 95 9.4 9.4 PG | ese 118 121

UAL 5.1 4.9 49 MBS 87 91 90

LAL ras 3.l 5.1 MVS 117 112 111

PAL 4.6 D7 +9 BLS 21 19 19

DLM (1) 1.4 1.3 1.4 TLP (i) (L/R) 9/9 9/9 9/9

DLM (11) 2.8 Dey Deh TLP (ii) (L/R) 13/12 ID 12/13

DLM (111) | 2.9 2D TLP (iti) (L/R) 18/18 19/18 17/18

DLM (iv) 3.3 3:1 3.2 TLP (iv) (L/R) 21/21 21/20 21/21

DLM (v) 2 2.4 2.4 TLP (v) (L/R) 16/16 15/16 15/15

TRL 14.6 23 12.0 FP (L/R) 15/14 - -

TW 6.3 6.1 6.2 PFS (L/R) 3/2 - -

TD 3.8 39 a IFS 10 - -

PEL Fel 6.9 69

TBL 6.2 6.1 6.1

HEL 6.2 6.3 672

DLP (1) 22, | ow)

DLP (11) 3.4 3:2 3.4

DLP (111) 3.8 3.8 3.7

DLP (iv) 4.2 4] 4.2

DLP (v) 3.6 35 3.6

TAL 39.4 36.5 34.7

TBW 3.8 3,5 3.4

TBD 3.1 vA, Sie

than the length of digit IV of the manus (OD 100.3% of

DLM IV); supraocular ridges not prominent; ear opening

very small (EL 6.0% of HL), deep, taller than wide, larger

than nostrils; single row of scales separates orbit from

supralabials; interorbital distance is narrow (IO 72.1% of

ES), shorter than head length (IO 33.0% of HL); eye to

Amphib. Reptile Conserv.

nostril distance slightly greater than eye to ear distance

(EN 102.1% of EE).

Dorsal surface of the trunk with smooth, small

homogeneous granules, 122 paravertebral granules;

117 smooth midventral scales; 87 midbody scales; 5/5

well developed tubercles on flanks; ventrolateral scales

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New species of Cnemaspis from Sri Lanka

slightly enlarged; granules on snout smooth and flattened,

larger than those on interorbital and occipital regions;

canthus rostralis not pronounced, 9/10 smooth oval

scales from eye to nostril; scales of interorbital region

oval and smooth; 2/2 weakly developed tubercles present

on sides of neck and around ear; ear opening vertically

oval, slanting from anterodorsal to posteroventral, 18/18

scales between anterior margin of the ear opening and

posterior margin of eye. Supralabials 8/8 and infralabials

9/8, becoming smaller towards the gape. Rostral scale

wider than long, partially divided (90%) by a median

groove and in contact with first supralabial. Nostrils

separated by 2/2 enlarged supranasals with one internasal

and 1/1 postnasal; no enlarged scales behind supranasals.

Nostrils oval, dorsolaterally oriented, not in contact with

first supralabials.

Mental subrhomboidal, as wide as long, posteriorly

in contact with three enlarged postmentals (smaller

than mental, and larger than chin scales); postmentals

in contact and bordered posteriorly by five smooth chin

scales (larger than nostrils), contact with the 1‘ and 2"4

infralabials; ventral scales smaller than chin scales,

and larger than nostrils. Smooth, rounded, juxtaposed

granule-like scales on chin and gular region; pectoral

and abdominal scales smooth, subimbricate towards

precloacal region, abdominal scales larger than dorsals;

21 belly scales across venter; smooth, subimbricate scales

around vent and base of tail; 15/14 femoral pores; 10

unpored interfemoral scales; 3/2 small posterior femoral

scales. Original tail of holotype longer than snout-vent

length (TAL 114.5% of SVL); hemipenial bulge greatly

swollen (TBW 3.8 mm), homogeneous scales on dorsal

aspect of tail directed posteriorly, 1/1 spine-like tubercles

present at base of tail, subcaudals very smooth; tail with

3-4 enlarged flattened obtuse scales forming whorls;

absence of post-cloacal spur on each side; smooth

subcaudals arranged into a median series of clearly

enlarged, hexagonal or subhexagonal scales.

Forelimbs moderately short, slender (LAL 15.1%

of SVL, UAL 14.8% of SVL) lower arm longer than

upper arm; hind limbs moderately long, tibia shorter

than femur (TBL 18.1% of SVL, FEL 20.5% of SVL).

Dorsal, anterior, ventral, and posterior surfaces of upper

arm with smooth scales, those on anterior surface twice

as large as those on other faces of limb; dorsal, anterior,

ventral, and posterior surfaces of lower arm with smooth

scales, those on posterior surface twice as large as

those of other parts; scales on dorsal surface of femur

smooth and granular, less imbricate scales on anterior,

posterior and ventral surfaces, scales on anterior surface

are twice the size of those of other aspects. All surfaces

of tibia with smooth scales; both anterior and posterior

surfaces of limbs bearing smooth granules, scales of the

ventral surface twice as large as those of other aspects.

Dorsal and ventral scales on the manus and the pes

smooth, granular; dorsal surfaces of digits with granular

scales. Digits elongate and slender with inflected distal

Amphib. Reptile Conserv.

phalanges, all bearing slightly recurved claws. Subdigital

lamellae entire (except divided at first interphalangial

joint), unnotched; total lamellae on manus (left/right):

digit I (11/11), digit IT (12/12), digit III (14/13), digit

IV (17/17), digit V (13/13); total lamellae on pes (left/

right): digit 1 (9/9), digit (13/12), digit IM (18/18), digit

IV (21/21), digit V (16/16); interdigital webbing absent;

length order of digits of manus (left): I (1.4 mm), V (2.5

mm), II (2.8 mm), II (3.1 mm), IV (3.3 mm); length

order of digits of pes (left): I (2.2 mm), II (3.4 mm), V

(3.6 mm), II (3.8 mm), IV (4.2 mm).

Variation of the type series. The SVL of adult speci-

mens in the type series (n = 3) and others (n = 5) ranges

from 30.3 to 34.4 mm, TAL ranges from 34.7—39.4 mm,

and TRL ranges from 12.0—14.6 mm; number of supral-

abials 8—9, and infralabials 8—9 (Table 1); spines on flank

3-5; interorbital scales 26—29; supraciliaries 9-11; can-

thal scales 8-10; scales from eye to tympanum 17-19;

total lamellae on digits of manus: digit I (10-11), digit

II (11-13), digit I (13-14), digit ITV (16-17), digit V

(12-13); total lamellae on digits of pes: digit I (12-13),

digit HI (17-19), digit [V (20-21), digit V (15-16); ven-

tral scales 111-117, midbody scales 87-91; paravertebral

granules 118—122; belly scales 19-21; unpored interfem-

oral scales 9-11 in males; femoral pores in males 14—15,

and unpored posterior femoral scales in males 2-3.

Color of living specimens. The body color on the dorsal

side is reddish brown; the dorsal head is randomly

scattered with black and white dots; a yellowish oval

patch on occiput, and a straight black middorsal dash

over midpoint of neck (Fig. 4); faded yellow patches

along vertebral midline; indistinct dark canthal stripe

extends through eye and above ear, terminating anterior

to forelimb insertion; the pupil of eye is circular and

black with the surrounding being golden brown; a series

of 4-5 mottled, irregular, dark brown transverse bands

with gray margins on dorsum of body; dorsum of tail

with 13—15 cinnamon brown blotches separating 12-14

faded dark brown bands; lateral view of labials and neck

consists of thin black dots in bright yellow background

like a zigzag mark; small dark spots (like eyes) present

on back side of femur; chin and gular with bright yellow,

vent and femur completely dirty yellow color.

Color of preserved specimens. Dorsum is light brown;

dorsum of head is randomly scattered with brown and

cream dots; an oval cream color patch on occiput, and

a straight dark brown middorsal dash over midpoint of

neck; a white post-orbital stripe present; labials with

black and cream spots; venter is completely dirty white;

tail with scattered markings on dorsal side.

Etymology. The specific epithet is an eponym Latinized

(anslemi) in the masculine genitive singular, honoring

the veteran Sri Lankan herpetologist Kongahage Anslem

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Karunarathna and Ukuwela

me i Fe oa See bd :

Fig. 4. Cnemaspis anslemi sp. nov. male holotype

(NMSL.2019.14.01) in life in-situ. (A) Dorsal view of the full

body displaying the typical color pattern and a straight black

middorsal dash over midpoint of neck, (B) Ventral aspect show-

ing gular and femoral colorations, (C) lateral view showing la-

bial coloration and zigzag pattern, (D) dorsal view of the full

body of female paratype (NMSL.2019.14.02) in life in-situ

from Udamaliboda, Samanala Nature Reserve, Sri Lanka. Pho-

tos: Kanishka Ukuwela and Suranjan Karunarathna.

Lawrence de Silva (the father of modern herpetology in

Sri Lanka) for his valuable contributions to Sri Lankan

herpetology and for inspiring the next generation of

herpetologists, including the authors.

Natural history. The lower Samanala Nature Reserve

area (along with Udamaliboda) comprises home gardens,

and tropical evergreen rainforests (Gunatileke and Guna-

tileke 1990) mixed with tea and rubber plantations. The

area comprises the Ratnapura and Kegalle districts and

lies between 6.759172° and 6.889842°N and 80.436194°

and 80.487717°E, at an elevation of 350-850 m. The

mean annual rainfall varies between 3,500 and 4,500

mm, received mostly via the southwest monsoon (May—

September). The mean annual temperature of the area

is 26.4—27.9 °C. Cnemaspis anslemi sp. nov. 1s a quite

rare species as six (+ 0.1) geckos per survey-hour were

found after covering a total area of 20 ha. This species

was restricted to tall straight trees with smooth bark and

thick canopy cover, and houses with tall clay walls with

crevices. These geckos could climb up to 7 m on vertical

surfaces of trees (Fig. 5). They were active during the day

Amphib. Reptile Conserv.

ees :

oki ae

maliboda, Samanala Nature Reserve, Kegalle District, Sri Lan-

ka. (A) Complete view of the forest hill, (B) shady forest with

thick leaf litter, (C) hundred years old house made using clay

and bricks, also with wattle and daub, (D) communal egg lay-

ing site on a clay wall. Photos: Madhava Botejue and Suranjan

Karunarathna.

time (08.00—17.00 h) and, when disturbed, sought ref-

uge in tree tops with crevices. The new species was sym-

patric (at local habitat scale) with several other geckos

(Cnemaspis samanalensis, Cnemaspis sp., Cyrtodactylus

triedrus, Cyrtodactylus sp., Gehyra mutilata, Hemidac-

tylus depressus, H. pieresii, H. frenatus, H. parvimacula-

tus, and Hemiphyllodactylus typus). The eggs were pure

white in color and almost spherical in shape (~5 mm),

with a slightly flattened side that attached to the clay-wall

substrate. This species has also been recorded from the

Lihinihela, Borangamuwa, and Warnagala areas in lower

Samanala Nature Reserve.

Conservation status. Application of the IUCN Red List

criteria indicates that C. anslemi sp. nov. is Critically

Endangered (CR) due to having an area of occupancy

(AOO) < 10 km? (six locations, 0.2 km? in total, assum-

ing a 100 m radius around the georeferenced locations)

and an extent of occurrence (EOO) < 100 km? (96.7 km?)

in the lower elevations of Central Province [Applicable

criteria are B2-b (i11)].

Remarks. Cnemaspis anslemi sp. nov. most closely

resembles C. gemunu, C. godagedarai, C. phillipsi, and

C. scalpensis. The type localities of these species are

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New species of Cnemaspis from Sri Lanka

Table 2. Key characters and identification features of 12 species belonging to the podihuna clade (scalpensis group), Sri Lanka. For

these species, the dorsal scales are homogeneous, ventral scales are smooth, and subcaudal scales are clearly enlarged, hexagonal,

or subhexagonal. Abbreviations: SVL—maximum snout to vent length in mm; SUP-supralabials; INF—infralabials; VEN—ventral

scales; BEL—belly scales; FSP—spines on the flank; MBO-midbody scales; PVT—paravertebral scales; UPF—unpored interfemoral

scales; FEP—femoral pores; LF4—lamellae on 4" finger; and LT4—lamellae on 4" toe.

FSP MBO PVT UPF FEP LF4 LT4

4-5 71-78 89-97 18-19 7-9 15-17-1721

3-4 87-9] 118-122 9-11 14-15 16-17 20-21

7-8 74-87 79-93 10-12 11-14 =15-17—- 18-19

5-6 98-102 101-106 8-9 12-13. 17-18 20-21

4-5 96-99 143-149 24-26 5-10 18-19 21-22

7-8 81-88 150-159 24-25 6-9 21-22 23-25

3-4 71-78 179-187 14-15 7-9 17-18 17-18

4-6 76-91 86-93 11-14 15-16 16-19 17-19

11-13 71-78 83-91 25-27 5-7 17-18 17-23

5-6 69-74 81-85 20-22 7-8 16-20 19-22

4-5 119-131 94-96 19-24 14-16 22-23 22-23

9-11 81-89 102-112 8-12 13-15 17-18 19-21

Species SVL SUP INF VEN BEL

C. alwisi 40.4 8-10 7-9 145-153 27-31

C. anslemi 34.4 8-9 8-9 111-117 19-21

C. gemunu 34.0 8-10 7-8 112-118 =13-16

C. godagedarai 35.5 7-8 7-8 133-137. 21-23

C. hitihami 41.7 8-9 7-9 132-135 21-22

C. kohukumburai 34.5 8-9 7-8 131-134 22-23

C. nilgala 32.9 7-8 6-7 122-129 17-19

C. phillipsi 36.6 8-9 8-9 128-143 =: 18-25

C. punctata 39.9 7-10 7-9 129-137 20-29

C. rajakarunai 40.2 8-9 9-1] 146-186 26-29

C. rammalensis 53.8 8-10 8-9 186-207 25-28

C. scalpensis 36.6 7-9 7-8 120-131 17-19

separated by ~38 km (Haggala in Nuwara Eliya), ~55

km (Ensalwatte in Deniyaya), ~83 km (Gammaduwa

in Matale), and ~47 km (Gannoruwa in Kandy) airline

distances, respectively, from Udamaliboda in Kegalle

(Fig. 1). Further, the new species can be distinguished

from C. gemunu, C. godagedarai, C. phillipsi, and C.

scalpensis by morphometric and meristic characters

(Table 2). We believe Cnemaspis cf. gemunu (AMB

7507, now in NMSL) collected from Borangamuwa in

Ratnapura District (6.742778°N, 80.707778°E; elevation

about 800 m) would most likely represent C. anslemi sp.

nov. according to the currently known distribution pattern

(see Agarwal et al. 2017). The records of Cnemaspis

scalpensis from Udamaliboda forest and vicinity by

Peabotuwage et al. (2012) also represent Cnemaspis

anslemi sp. nov.

Discussion

The discovery and description of a novel species here

adds yet another member to this speciose genus, increas-

ing the known diversity of Cnemaspis in Sri Lanka to 33

species, all of which are endemic to the island. Several

new descriptions during the last decade (e.g., Bauer et

al. 2007; Manamendra-Arachchi et al. 2007; Wickrama-

singhe and Munindradasa 2007; Vidanapathirana et al.

2014; Wickramasinghe et al. 2016; Batuwita and Udu-

gampala 2017; Batuwita et al. 2019; de Silva et al. 2019;

Karunarathna et al. 2019a,b) have greatly advanced our

knowledge on the diversity of these diminutive day geck-

Os, Increasing the total diversity from just four species.

This trend most likely suggests that the diversity of Sri

Lankan Cnemaspis is still underestimated, and further

studies would most likely reveal more species from the

varied natural and semi-natural habitats of Sri Lanka.

Although Sri Lankan Cnemaspis are likely derived from

the Indian radiation, the current diversity of this genus 1s

Amphib. Reptile Conserv.

probably the result of multiple colonization events (poly-

phyletic origin) as opposed to a single in situ radiation

(monophyly); however, these phylogenetic and biogeo-

graphic affinities have yet to be confirmed (Agarwal et

al. 2017; Bauer et al. 2007; Karunarathna et al. 2019b).

We tentatively assign this species to the podihuna clade

on the basis of clearly enlarged, hexagonal, or subhex-

agonal subcaudal scales (Fig. 6).

The preliminary studies reported here indicate that

this novel species is frequently found in home garden

habitats, as opposed to natural forest habitats, in the

Udamaliboda region (Samanala Nature Reserve). Dur-

ing the survey from 2006 to 2019, only five specimens

were found in the natural forest habitats of the Samanala

Nature Reserve. The home gardens in which the new

Species was observed are heavily shaded and humid. The

low encounter rates in natural forests could also be due to

the low visibility conditions caused by the dense canopy,

and this species may possibly occupy higher perches on

the tree trunks, thus avoiding detection. However, fur-

ther studies are necessary to ascertain this fact. Like most

of the Sri Lankan Cnemaspis known so far (Bauer et al.

2007; Agarwal et al. 2017), the new species has a very

small range, most likely due to the narrow ecological

niche of this species (Slatyer et al. 2013). Its small size

may reduce both dispersal ability as well as ecological

tolerance levels. The Udamaliboda trail of Samanala Na-

ture Reserve 1s inhabited by 11 species of geckos, includ-

ing two undescribed species, two Critically Endangered,

two Endangered, and two Vulnerable species (Peabotu-

wage et al. 2012). Overall, 65 reptile species have already

being recorded from the type locality, indicating that the

Udamaliboda 1s a local hotspot of reptile diversity.

The type locality of this species and the surrounding

areas are currently subjected to encroachment via tea cul-

tivation and mini-hydropower projects. Such activities

will certainly reduce the crucial natural and semi-natural

September 2019 | Volume 13 | Number 2 | e187

Karunarathna and Ukuwela

Fig. 6. Morphological characters that differentiate the species of podihuna and kandiana clades. (A) Scales and pores around vent

in podihuna clade, (B) scales and pores around vent in kandiana clade, (C) keeled and imbricate belly scales, (D) smooth and imbri-

cate belly scales, (E) heterogeneous keeled dorsal granules, (F) homogeneous smooth dorsal granules, (G) smooth sub-hexagonal

subcaudals in podihuna clade, (H) smooth, solid hexagonal subcaudals in podihuna clade, (1) smooth, small, and irregular subcau-

dals in kandiana clade, (J) keeled, small, and irregular subcaudals in kandiana clade (yellow arrows: unpored posterior femoral

scales in males; blue arrows: femoral pores in males; green arrows: unpored anterior femoral scales in males; red arrows: precloacal

pores in males; white arrows: unpored interfemoral scales in males).

habitats of this range-restricted species. Thus, authorities rare or even undescribed species is minimized.

should carefully consider new proposals for mini-hydro-

power plants near or within natural habitats like these,in Acknowledgements.—We thank Chandana Sooriyaban-

order to ensure that loss of the habitats occupied by such _—_ dara (The Director General of DWC), Laxman Peiris

Amphib. Reptile Conserv. 23 September 2019 | Volume 13 | Number 2 | e187

New species of Cnemaspis from Sri Lanka

(Deputy Director — Research division of DWC), the re-

search committee, and the field staff of the Department

of Wildlife (WL/3/2/1/14/12 and WL/3/2/42/18 a and b)

and Conservator General of Forest Department (FRC/5,

and FRC/6) for granting permission and the field staff

for assisting during the field surveys. Nanda Wickrama-

singhe, Sanuja Kasthuriarachchi, Lankani Somaratne,

Chandrika Munasinghe, Rasika Dasanayake, Ravindra

Wickramanayake, and P. Gunasiri at NMSL assisted

while we were examining collections under their care.

Anslem de Silva, Aaron Bauer, Thilina Surasinghe, Bud-

dhika Madurapperuma, Chamara Amarasinghe, Tharaka

Kusuminda, Indika Peabotuwage, Nirmala Perera, Mad-

hava Botejue, Dinesh Gabadage, Hasantha Wiyethunga,

D.M. Karunarathna, Kawmini Karunarathna, Rashmini

Karunarathna, Thesanya Karunarathna, and Niranjan

Karunarathna provided valuable assistance in numerous

stages of this study. This work was mainly supported by a

Nagao Natural Environment Foundation (2018-20) grant

to SK. Finally, we would like to thank the anonymous

reviewers for constructive comments that helped to im-

prove the manuscript.

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pattern. Ecology Letters 16: 1,104—1,114.

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ka: a Colour Guide with Field Keys. Chimaira, Frank-

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Vidanapathirana DR, Rajeev MDG, Wickramasinghe

N, Fernando SS, Wickramasinghe LJM. 2014. Cne-

maspis rammalensis sp. nov., Sri Lanka’s largest day-

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malakanda Man and Biosphere Reserve in southern

Sri Lanka. Zootaxa 3755: 273-286.

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view of the genus Cnemaspis Strauch, 1887 (Sauria:

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new species. Zootaxa 1490: 1-63.

Wickramasinghe LJM, Vidanapathirana DR, Rathnayake

RMGBP. 2016. Cnemaspis rajakarunai sp. nov., a rock

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pis) from Salgala, an unprotected lowland rainforest

in Sri Lanka. Zootaxa 4168: 92-108.

Suranjan Karunarathna obtained his Masters in Environmental Management at the University

of Colombo, Sri Lanka. His scientific exploration of biodiversity began with the Young Zoologists’

Association of Sri Lanka (YZA) in early 2000, and he served as president of YZA in 2007. As a

wildlife researcher, Suranjan conducts research on herpetofaunal ecology and taxonomy, and he

also promotes the scientific basis for raising awareness on the importance of biodiversity and its

conservation among the Sri Lankan community. He is an active member of many specialist groups

in the IUCN/SSC, and an expert committee member for herpetofauna in the National Red List

development programs, Sri Lanka, since 2004. Photo: Lark Hayes.

Kanishka D.B. Ukuwela is currently a Senior Lecturer in Zoology at the Rajarata University

of Sri Lanka. He holds a B.S. (Hons.) degree in Zoology from the University of Peradeniya, Sri

Lanka and a Ph.D. in Evolutionary Biology from the University of Adelaide, Australia. His current

research is focused on the origins, evolution, systematics, and conservation of the South Asian

herpetofauna. Photo: [suri Jayawardena.

September 2019 | Volume 13 | Number 2 | e187

New species of Cnemaspis from Sri Lanka

Appendix 1.

Comparative material:

Cnemaspis alwisi. NMSL 2004.09.01 (holotype), NMSL 2004.09.02 (paratype), NMSL 2004.09.03 (paratype), WHT

5918, WHT 6518, WHT 6519, WHT 7336, WHT 7337, WHT 7338, WHT 7343, WHT 7344, WHT 7345, WHT 7346.

C. amith: BMNH 63.3.19.1066A (holotype), BMNH 63.3.19.1066B (paratype), BMNH 63.3.19.1066C (paratype).

C. butewai: NMSL 2019.07.01 (holotype), NMSL 2019.07.02 (paratype), NMSL 2019.07.03 (paratype).

C. gemunu: AMB 7495 (holotype), AMB 7507 (paratype?), WHT 7221, WHT 7347, WHT 7348, NMSL 2006.11.01,

NMSL 2006.11.02, NMSL 2006.11.03, NMSL 2006.11.04.

C. godagedarai: NMSL 2019.09.01 (holotype), NMSL 2019.16.01 (paratype), NMSL 2019.16.02 (paratype).

C. gotaimbarai: NMSL 2019.04.01 (holotype), NMSL 2019.04.02 (paratype), NMSL 2019.04.03 (paratype).

C. hitihami. NMSL 2019.06.01 (holotype), NMSL 2019.06.02 (paratype), NMSL 2019.06.03 (paratype).

C. ingerorum. WHT 7332 (holotype), WHT 7330 (paratype), WHT 7331 (paratype).

C. kallima: WHT 7245 (holotype), WHT 7222 (paratype), WHT 7227 (paratype), WHT 7228 (paratype), WHT 7229

(paratype), WHT 7230(paratype), WHT 7239 (paratype), WHT 7249 (paratype), WHT 7251 (paratype), WHT 7252

(paratype), WHT 7253 (paratype), WHT 7254 (paratype), WHT 7255 (paratype).

C. kandambyi. WHT 9466 (holotype), WHT 9467 (paratype).

C. kandiana: BMNH 53.4.1.1 (lectotype), BMNH 80.2.2.119A (paralectotype), BMNH 80.2.2.119B (paralectotype),

BMNH 80.2.2.119C (paralectotype), WHT 7212, WHT 7213, WHT 7267, WHT 7305, WHT 7307, WHT 7308, WHT

7310, WHT 7313, WHT 7319, WHT 7322.

C. kivulegedarai: NMSL 2019.08.01 (holotype), NMSL 2019.08.02 (paratype), NMSL 2019.08.03 (paratype).

C. kohukumburai: NMSL 2019.05.01 (holotype), NMSL 2019.05.02 (paratype), NMSL 2019.05.03 (paratype).

C. kumarasinghei: NMSL 2006.13.01 (holotype), NMSL 2006.13.02 (paratype).

C. latha: WHT 7214 (holotype).

C. menikay: WHT 7219 (holotype), WHT 7218 (paratype), WHT 7349 (paratype).

C. molligodai: NMSL 2006.14.01 (holotype), NMSL 2006.14.02 (paratype), NMSL 2006.14.03 (paratype), NMSL

2006.14.04 (paratype), NMSL 2006.14.05 (paratype).

C. nandimithrai: NMSL 2019.01.01 (holotype), NMSL 2019.01.02 (paratype), NMSL 2019.01.03 (paratype).

C. nilgala: NMSL 2018.07.01 (holotype), NMSL 2018.06.01 (paratype), NMSL 2018.06.02 (paratype), NMSL

2018.06.03 (paratype).

C. pava: WHT 7286 (holotype), WHT 7281 (paratype), WHT 7282 (paratype), WHT 7283 (paratype), WHT 7285

(paratype), WHT 7288 (paratype), WHT 7289 (paratype), WHT 7290 (paratype), WHT 7291 (paratype), WHT 7292

(paratype), WHT 7293 (paratype), WHT 7294 (paratype), WHT 7295 (paratype), WHT 7296 (paratype), WHT 7297

(paratype), WHT 7298 (paratype), WHT 7299 (paratype), WHT 7300 (paratype), WHT 7301 (paratype), WHT 7302

(paratype).

C. phillipsi. WHT 7248 (holotype), WHT 7236 (paratype), WHT 7237 (paratype), WHT 7238 (paratype).

Amphib. Reptile Conserv. 26 September 2019 | Volume 13 | Number 2 | e187

Karunarathna and Ukuwela

C. podihuna. BMNH 1946.8.1.20 (holotype), NMSL 2006.10.02, NMSL 2006.10.03, NMSL 2006.10.04.

C. pulchra: WHT 7023 (holotype), WHT 1573a (paratype), WHT 7011 (paratype), WHT 7021 (paratype), WHT 7022

(paratype).

C. punctata. WHT 7256 (holotype), WHT 7223 (paratype), WHT 7226 (paratype), WHT 7243 (paratype), WHT 7244

(paratype).

C. rajakarunai: NMSL 2016.07.01 (holotype), DWC 2016.05.01 (paratype), DWC 2016.05.02 (paratype).

C. rammalensis: NMSL 2013.25.01 (holotype), DWC 2013.05.001.

C. retigalensis: NMSL 2006.12.01 (holotype), NMSL 2006.12.02 (paratype), NMSL 2006.12.03 (paratype), NMSL

2006.12.04 (paratype).

C. samanalensis: NMSL 2006.15.01 (holotype), NMSL 2006.15.02 (paratype), NMSL 2006.15.03 (paratype), NMSL

2006.15.04 (paratype), NMSL 2006.15.05 (paratype).

C. scalpensis: NMSL 2004.01.01 (neotype), NMSL 2004.02.01, NMSL 2004.03.01, NMSL 2004.04.01, WHT 7265,

WHT 7268, WHT 7269, WHT 7274, WHT 7275, WHT 7276, WHT 7320.

C. silvula: WHT 7208 (holotype), WHT 7206 (paratype), WHT 7207 (paratype), WHT 7209 (paratype), WHT 7210

(paratype), WHT 7216 (paratype), WHT 7217 (paratype), WHT 7018, WHT 7027, WHT 7202, WHT 7203, WHT

7220, WHT 7354, WHT 7333.

C. tropidogater. BMNH 71.12.14.49 (lectotype), NMSL 5152, NMSL 5151, NMSL 5159, NMSL 5157, NMSL 5970,

NMSL 5974.

C. upendrai: WHT 7189 (holotype), WHT 7184 (paratype), WHT 7187 (paratype), WHT 7188 (paratype), WHT 7181

(paratype), WHT 7182 (paratype), WHT 7183 (paratype), WHT 7185 (paratype), WHT 7190 (paratype), WHT 7191

(paratype), WHT 7192 (paratype), WHT 7193 (paratype), WHT 7194 (paratype), WHT 7195 (paratype), WHT 7196

(paratype), WHT 7197 (paratype), WHT 7260 (paratype).

Amphib. Reptile Conserv. 27 September 2019 | Volume 13 | Number 2 | e187

Official journal website:

amphibian-reptile-conservation.org

Amphibian & Reptile Conservation

13(2) [General Section]: 28-30 (e188).

Book Review

How Snakes Work: Structure, Function

and Behavior of the World’s Snakes

Brian I. Crother

Department of Biological Sciences, Southeastern Louisiana University, Hammond, Louisiana, USA

Keywords. Reptilia, Squamata, Ophidia, anatomy, locomotion, physiology, reproduction, thermoregulation

Citation: Crother BI. 2019. Book Review—How Snakes Work: Structure, Function and Behavior of the World’s Snakes. Amphibian & Reptile

Conservation 13(2) [General Section]: 28—30 (e188).

ternational (CC BY 4.0): https://creativecommons.org/licenses/by/4.0/], which permits unrestricted use, distribution, and reproduction in any medium,

provided the original author and source are credited. The official and authorized publication credit sources, which will be duly enforced, are as follows:

Official journal title Amphibian & Reptile Conservation; official journal website: amphibian-reptile-conservation.org.

Received: 26 August 2019; Accepted: 29 August 2019; Published: 12 September 2019

“They are magnificent. They are splendid creations

ingeniously constructed after eons of gradual

change.” Kauffeld, Snakes and Snake Hunting, 1957

There are many books about snakes for every level of

understanding, but rarely, if ever, has there been a book

about snakes that matches Harvey Lillywhite’s offering.

As I went through the volume, I kept thinking how I wish

I had such a book when I was in high school/college, or

even as akid. I wrote a term paper in high school on how

snakes move and this book has an entire chapter devoted

to it! I honestly got goosebumps and bursts of adrenaline

as I read it, because it brought me back to my younger

days of discovery about the biology of snakes. How

fascinating they were/are! Lillywhite’s volume details all

the reasons why snake are magnificent organisms.

The volume is in a comfortable 8 1/2 by 11-inch

format with 241 pages, glossary, index, and references at

the end of each chapter. Rick Shine wrote the Foreword

and Lillywhite explains in a short Preface his personal

reasons and goals for the book. Lillywhite hoped that he

“might bring a unique perspective to the subject matter,

and to cover a range of topics...that will interest a wide

group of readers.” I think he is completely successful

in reaching those goals. Perhaps the reason he is so

successful—and why this is such a terrific book—is

because the author is an insider, a professional research

scientist who has studied how snakes work for his entire

career. We are all lucky that Lillywhite is sharing his

expertise and passion with us in such a volume.

There are nine chapters: 1) Evolutionary History and

Classification, 2) Feeding, Digestion, and Water Balance,

3) Locomotion: How Snakes Move, 4) Temperature

Correspondence. bcrother@southeastern.edu

Amphib. Reptile Conserv.

and Ectothermy, 5) Structure and Function of Skin,

6) Internal Transport: Circulation and Respiration,

7) Perceiving the Snake’s World: Structure and

Function of Sense Organs, 8) Sound Production, and 9)

Courtship and Reproduction. There is obviously a lot of

information here and it is as up to date as it could be.

Each of the chapters is wonderfully adorned with terrific

photos and illustrations that help the reader grasp and/or

visualize the text descriptions.

Even in a book on how snakes work, a chapter on

evolution and classification is practically obligatory

because it puts the rest of the book’s information into

context. Maybe the most important nugget of knowledge

given in this chapter is “The latter [snakes] are essentially

highly derived lizards.” It is critical to point out that

snakes are not related to lizards, but are lizards. That

realization makes all the novel evolutionary changes

relative to lizards that are described in this book even

more remarkable. With that fact written in the book, it

is somewhat unfortunate that an accompanying figure

shows snakes and lizards as separate branches on a

phylogeny of the tetrapods. The natural history comments

in the “tip toe through the taxa” section of the chapter are

a welcome addition.

An example of how having a volume written by a

professional research scientist enhances the content

is near the beginning of Chapter 2. Lillywhite wrote,

“The following discussion 1s essentially speculative,

but it provides a conceptual basis from which to launch

considerations of feeding and digestion in snakes.”

Because of his long history of work in the field, his

speculations are valuable and make you feel like you

just asked an expert a question and he is giving you

September 2019 | Volume 13 | Number 2 | e188

Crother

his unique insight. I consider it a bonus. Beyond the

speculation, this chapter 1s a trove of information on the

diversity of feeding adaptations among snakes, such as

the physiology required to handle digestion of various

food types, and the myriad aspects of how snakes

physiologically deal with water.

So, you think you know how snakes move? In my

high school term paper that I previously mentioned, I

described three modes: lateral, concertina, and rectilinear

motion. I was woefully incomplete. The illustrations in

Harvey’s book, from anatomy to movement, are excellent

and useful. The final section of this chapter, titled Snakes

as Robots, is a treat.

Of course, we recognize snakes as ectotherms, but

as Lillywhite reveals in Chapter 4 on “Temperature

and Ectothermy,” being an ectotherm is an extremely

complicated deal for snakes. The chapter goes through

these many complications and even provides examples

of endothermy in snakes, which makes the label

“ectotherm” require an even more broad interpretation.

Lillywhite does a great job pointing out how much is not

known about thermoregulation, especially with regard

to trade-offs or cost-benefits when thermoregulation is

coupled with any of the myriad of other physiological

processes of the organism.

There is something magical about seeing snake skin

that is either as glossy as glass, like mud snakes (Farancia

abacura), or spectacularly iridescent, like rainbow boas

(Epicrates cenchria). Well the chapter on skin covers it

all and tells you everything you need (or wanted) to know

about snake skin, from cultural references to histology

and micrographs to mimicry and color/pattern variation.

As usual for this book, the photos and illustrations are

terrific!

The concluding paragraph of the chapter on Internal

Transport opens with: “Few persons probably give

much thought to how the blood circulates within a snake

and how the lung functions to assist in the delivery of

oxygen.” I have to agree with Lillywhite here, but I also

have to add that after reading the chapter one can’t help

but learn things, even if you think you already know

everything there is to know about circulation and gas

exchange in snakes.

One of the most amazing aspects of snake biology

is the way they perceive the environment around them.

Snakes are famous for their chemosensory abilities, and

some groups of snakes are well-known for their thermal

and infrared radiation detection abilities. The chapter

on how snakes perceive the world covers those topics

in excellent fashion. Even though hearing is less of an

obvious pathway for snakes in sensing the environment,

the coverage on that is well done here too. When I teach

herpetology, I love to talk about snake eyes because of all

the novel structures (and of the typical structures that are

absent), because it seems fairly well-established that in

the evolution of snakes the eyes were radically reduced if

not essentially lost.

Amphib. Reptile Conserv.

STRUCTURE, FUNCTION

and BEHAVIOR of the

WORLD'S SNAKES

Title: How Snakes Work: Structure, Function and Behavior

of the World’s Snakes

Author: Harvey B. Lillywhite

ISBN: 978-0-19-538037-8

Publisher: Oxford University Press

Pages: xiii + 241; Price: $61 (USD)

Thus, I was disappointed to see that all those

interesting things were not pointed out in the discussion

of snake eyes in the book. The illustration of the eye

was simplistic and sort of generic. For example, there

are no muscles associated with the ciliary body in snake

eyes, and the conus, sphincter, and dilatator all have

mesodermal origins in snakes, but are ectodermal (!) in

lizards. Those are big differences.

Snake vision received the same generic treatment. For

example, regarding pupil shape, while it's true that a slit

pupil can close more completely than a round pupil, which

the author attributes to crepuscular and nocturnal activity,

that's not the primary function. There are lots of diurnal

snakes with slit pupils. The real function 1s that all animals

with a slit pupil also have a multifocal lens. The slit allows

a reduction in light while still allowing all of that light to

pass through all peripheral layers of the lens (which have

different refractive indices or color filters), rather than just

through the lens center. No doubt, almost every aspect

of snake vision is poorly known, but what is known is

spectacular and its coverage in greater detail would have

fit perfectly well with the rest of this excellent book.

September 2019 | Volume 13 | Number 2 | e188

Book Review: How Snakes Work

When people think of snakes, how many first

think of sound production? Perhaps folks who mainly

encounter rattlesnakes during their lives may associate

snakes with sounds, but typically snakes are associated

with their stealth, with their silence. And that is what

makes the chapter on sound so enjoyable. I want to

hear a gopher snake bellow! I want to hear a cloacal

pop or a king cobra growl! Like vision, there is much

that is not known about snake sound production and

communication, and the author does a nice job bringing

up those questions.

The beginning and end of lineages is intimately tied

to successful reproduction, and the book closes with

a chapter on the subject. It is another very well-done

Amphib. Reptile Conserv.

chapter, with cultural and biological information as well

as bits of expert-based speculation included. I greatly

suspect that for many non-experts this chapter will hold

a number of surprises about snakes, which really, in a

nutshell, is what is great about this book. I very much

like the encouraging, upbeat style of writing that gets the

reader excited about the topics.

I think this is a terrific book for readers with a variety

of knowledge levels. I really believe elementary school

aged kids can get excited by this book and grow with the

book, appreciating it more and more as they learn and

progress through school. I also think the book fits well at

the university level and would be a welcome addition to

any biologist’s library, including herpetologists!

Brian I. Crother is the Schlieder Foundation Professor of Biological Sciences

and interim Department Head of Computer Sciences at Southeastern Louisiana

University in Hammond, Louisiana, USA. Brian earned his B.S. from California

State University at Dominguez Hills, his Ph.D. from the University of Miami,

Florida, USA, and he conducted post-doctoral research at the University of

Texas, Austin. He has well over 100 publications on a broad range of topics,

including edited books on Caribbean Amphibians and Reptiles, Ecology and

Evolution in the Tropics: A Herpetological Perspective and Assumptions that

Inhibit Progress in Comparative Biology. Brian was the chair and coauthor

of the 5" through 8" editions of the Scientific and Standard English Names of

Amphibians and Reptiles of North America North of Mexico. He is active in

several professional organizations and is an ex-president of both the Society for

the Study of Amphibians and Reptiles and the American Society of Ichthyologists

and Herpetologists. Brian’s research interests are broad, but have in common

that they cover amphibians, reptiles, and/or evolution (empirical, theoretical,

and philosophical).

September 2019 | Volume 13 | Number 2 | e188

Official journal website:

amphibian-reptile-conservation.org

Amphibian & Reptile Conservation

13(2) [General Section]: 31-94 (e189).

The herpetofauna of Coahuila, Mexico: composition,

distribution, and conservation status

‘David Lazcano, ‘Manuel Nevarez-de los Reyes, *Eli Garcia-Padilla, "Jerry D. Johnson,

3Vicente Mata-Silva, 7Dominic L. DeSantis, and *°*Larry David Wilson

‘Universidad Autonoma de Nuevo Leon, Facultad de Ciencias Biolégicas, Laboratorio de Herpetologia, Apartado Postal 157, San Nicolas de los

Garza, Nuevo Leon, C.P. 66450, MEXICO ?Oaxaca de Juarez, Oaxaca 68023, MEXICO *Department of Biological Sciences, The University of

Texas at El Paso, El Paso, Texas 79968-0500, USA “Centro Zamorano de Biodiversidad, Escuela Agricola Panamericana Zamorano, Departamento

de Francisco Morazadn, HONDURAS °1350 Pelican Court, Homestead, Florida 33035, USA

Abstract.—The herpetofauna of Coahuila, Mexico, is comprised of 143 species, including 20 anurans, four

caudates, 106 squamates, and 13 turtles. The number of species documented among the 10 physiographic

regions recognized ranges from 38 in the Laguna de Mayran to 91 in the Sierras y Llanuras Coahuilenses. The

individual species occupy from one to 10 regions (xX = 3.5). The numbers of species that occupy individual

regions range from 23 in the Sierras y Llanuras Coahuilenses to only one in each of three different regions. A

Coefficient of Biogeographic Resemblance (CBR) matrix indicates numbers of shared species among the 10

physiographic regions ranging from 20 between Llanuras de Coahuila y Nuevo Leon and Gran Sierra Plegada

to 45 between Serranias del Burro and Sierras y Llanuras Coahuilenses. A similarity dendrogram based on the

Unweighted Pair Group Method with Arithmetic Averages (UPGMA) reveals that the Llanuras de Coahuila y

Nuevo Leon region is most dissimilar when compared to the other nine regions in Coahuila (48.0 % similarity);

all nine other regions cluster together at 57.0% and the highest similarity is 92.0% between Laguna de Mayran

and Sierra de la Paila. The distribution patterns concerning numbers of shared species reflect higher similarity

between regions that share geographic contact with each other and have comparable ecological parameters.

The percentage of species restricted to one or two physiographic regions is 63.6%, indicating a moderately

narrow distribution for many species. The largest number of species is placed in the non-endemic category

(100), followed by the country endemics (31), state endemics (nine), and non-natives (three). The principal

environmental threats to the herpetofauna are urban development, industrial pollution, deforestation, road

effects, mining and energy projects, natural gas fracking, wind turbines, elimination due to cultural beliefs

and practices, commercial trade, and forest fires. The conservation status of the native species is assessed

by using the SEMARNAT (NOM-59), IUCN, and EVS systems, of which the EVS system proved to be the most

useful. The EVS rankings also were used to determine how species in the IUCN categories of Not Evaluated (NE)

and Least Concern (LC) might be evaluated more informatively. Using the Relative Herpetofaunal Priority (RHP)

methodology, we determined that the most significant herpetofaunas are those of the Gran Sierra Plegada and

the Sierras y Llanuras Coahuilenses. Nineteen protected areas are established in Coahuila and we predict

that 119 of the 143 species in the state occur in them, based on their respective physiographic distributions.

Finally, a set of conclusions and recommendations for the future protection of the Coahuilan herpetofauna is

presented.

Key words. Amphibians, anurans, caudates, physiographic regions, protected areas, protection recommendation,

reptiles, squamates, turtles

Resumen.—La herpetofauna de Coahuila, México consiste de 143 especies, incluyendo 20 anuros, cuatro

caudados, 106 escamosos, y 13 tortugas. El numero de especies documentadas entre las 10 regiones

fisiograficas reconocidas va de 38 en la Laguna de Mayran, a 91 en Sierras y Llanuras Coahuilenses. Las especies

individuales ocupan de una a 10 regiones (Xx = 3.5). El mayor numero de especies en una sola region va de 23 en

la Sierras y Llanuras Coahuilenses a una en cada una de las tres regiones. Una matriz de coeficiente de similitud

biogeografica (CSB) indica que el numero de especies compartidas entre las 10 regiones fisiograficas va de

20 entre Llanuras de Coahuila y Nuevo Leon y la Gran Sierra Plegada a 45 entre Serranias del Burro y Sierras

y Llanuras Coahuilenses. Un dendrograma de similitud basado en el Método por Agrupamiento de Pares no

Ponderado con Media Aritmetica (MAPMA) revela que sobre una base jerarquica, Llanuras de Coahuila y Nuevo

Leon es la mas desigual cuando se le compara con las otras nueve regiones en Coahuila (48.0% similitud);

todas las nueve regiones se agrupan en 57.0% y la mayor similitud es 92,0% entre Laguna de Mayran y Sierra

de la Paila. Los patrones de distribucion con respecto al numero de especies compartidas reflejan una mayor

similitud entre las regiones en contacto geografico y con parametros ecologicos comparables. El porcentaje

Correspondence. imantodes52@hotmail.com (DL), digitostigma@gmail.com (MNR), eligarcia_18@hotmail.com (EGP), jjohnson@utep.edu

(JDJ), vmata@utep.edu (VMS), didesantis@miners.utep.edu (DLD), bufodoc@aol.com (*LDW)

Amphib. Reptile Conserv. 31 October 2019 | Volume 13 | Number 2 | e189

The herpetofauna of Coahuila, Mexico

de especies restringidas a una o dos regiones fisiograficas es de 63.6%, indicando una distribucion moderada

para muchas especies. El mayor numero de especies esta ubicado en la categoria de no endemica (100),

seguido de endémicas al pais (31), endémicas al estado (nueve), y no nativas (tres). Las principales amenazas

ambientales a la herpetofauna son el desarrollo urbano, contaminacion industrial, deforestacion, efectos de

carreteras, actividad minera, actividad petrolera (fracking), turbinas edlicas, matanza por falta de educacion y

para uso medicinal, colecta y comercio, e incendios forestales. Calculamos el estatus de conservacion de las

especies nativas usando los sistemas de SEMARNAT (NOM-059), UICN, y el EVS, de los cuales el EVS resulto

ser mas util. También usamos los rangos de EVS para determinar como las especies en las categorias de No

Evaluada (NE) y de Preocupacion Menor (PM) de la UICN podrian ser evaluadas de una forma mas informativa.

Asimismo, usando el método de Prioridad Herpetofaunistica Relativa (PHR), determinamos que la herpetofauna

mas significativa es la de Gran Sierra Plegada y la de Sierras y Llanuras Coahuilenses. Diecinueve areas

protegidas han sido establecidas en Coahuila y predecimos que 119 de las 143 especies que ocurren en el

estado seran encontradas en estas areas, basado en su distribuciOn geografica. Finalmente, incluimos un

grupo de conclusiones y recomendaciones para la futura proteccion de la herpetofauna de Coahuila.

Palabras claves. Anfibios, anuros, caudados, regiones fisiograficas, areas protegidas, recomendaciones de proteccion,

reptiles, escamosos, tortugas

Citation: Lazcano D, Nevarez-de los Reyes M, Garcia-Padilla E, Johnson JD, Mata-Silva V, DeSantis DL, Wilson LD. 2019. The herpetofauna of

Coahuila, Mexico: composition, distribution, and conservation status. Amphibian & Reptile Conservation 13(2) [General Section]: 31-94 (e189).

4.0 International (CC BY 4.0): https://creativecommons.org/licenses/by/4.0/], which permits unrestricted use, distribution, and reproduction in any

medium, provided the original author and source are credited. The official and authorized publication credit sources, which will be duly enforced, are

as follows: official journal title Amphibian & Reptile Conservation; official journal website: amphibian-reptile-conservation.org.

Received: 26 January 2019; Accepted: 20 July 2019; Published: 9 October 2019

“Like it or not, and prepared or not, we are the mind Coahuila y Nuevo Leon region and the southeastern

and stewards of the living world. Our own ultimate future corner in which is found a small portion of the

depends upon that understanding.” Gran Sierra Plegada region (Fig. 1). In Mexico, the

Chihuahuan Desert also encompasses “a large portion

E. O. WILSon (2016) of the state of Chihuahua..., northeastern Durango, the

extreme northern part of Zacatecas, and small western

Introduction portions of Nuevo Leon” (https://www.worldwildlife.

org/ecoregions/nal303; accessed 24 December 2017).

Coahuila is the third largest state of Mexico after The purpose of this paper, similar to that of the others

Chihuahua and Sonora, all of which border the United — in the Mexican Conservation Series (see below), is to

States of America. Coahuila is bounded to the north by document the composition, physiographic distribution,

the US state of Texas, to the west by the Mexican states —_ and conservation status of the herpetofauna of Coahuila.

of Chihuahua and Durango, to the south by Zacatecas _In general, the format of the earlier papers in this series

and a small portion of San Louis Potosi, and to the east —_1s followed here.

by Nuevo Leon. Coahuila encompasses 151,595 km?

and falls in size between the US states of Illinois and Materials and Methods

Georgia (http://cuentame.inegi.gob.mx/monografias/

informacion/coah/default.aspx?tema=me&e=05; Our Taxonomic Position

accessed 26 August 2017). Coahuila’s population was

2,954,915 in 2015, which is 2.5% of the same-year In this paper, we follow the same taxonomic position

estimate for the entire country of Mexico (119,530,753). as explained in previous works on other portions of

Coahuila is one of the six least densely populated states | Mesoamerica (Johnson et al. 2015a,b; Mata-Silva et al.

in Mexico (the others are Chihuahua, Sonora, Campeche, 2015). Johnson (2015b) can be consulted for a statement

Durango, and Baja California Sur), each of which has __ of this position, with special reference to the subspecies

fewer than 20 inhabitants per square kilometer; the — concept.

figure for Coahuila is 19.5 (http://cuentame.ineg1.

gob.mx/monografias/informacion/coah/default. | Updating the Herpetofaunal List

aspx?tema=meé&e=05; accessed 26 August 2017). The

capital and largest city in Coahuila is Saltillo, located in = Several recent works on the herpetofauna of Coahuila

the southeastern portion of the state. are available. Lemos-Espinal and Smith (2007) created

Much of Coahuila lies within the borders of the a bilingual (Spanish and English) treatment of the state

Chihuahuan Desert (Lazcano et al. 2017), except for § herpetofauna, in which they recognized 129 species.

the northeastern portion located within the Llanuras de —__ Eight years later, Lemos-Espinal et al. (2015) compiled

Amphib. Reptile Conserv. 32 October 2019 | Volume 13 | Number 2 | e189

Lazcano et al.

28°30'N

27°0'N

25°30'N

') BDM:

m™ GSP:

mI LCN:

mm LGM:

mi LSV:

mm SLC:

mi SLP:

STR:

m= SDB:

mu PSP:

TOPOGRAPHIC ASPECTS

Roads

Municipality boundaries

L_] State boundaries

4102700

PHYSIOGRAPHIC REGIONS

BOLSON DE MAPIMIi

GRAN SIERRA PLEGADA

LLANURAS DE COAHUILA Y NUEVO LEON

LAGUNA DE MAYRAN

LLANURAS Y SIERRAS VOLCANICAS

SIERRAS Y LLANURAS COAHUILENSES

SIERRA DE LA PAILA

SIERRAS TRANSVERSALES

SERRANIAS DEL BURRO

PLIEGUES SALTILLO - PARRAS

Fig. 1. Physiographic regions of Coahuila, Mexico. Abbreviations are as follows: BDM = Bolson de Mapimi; GSP = Gran Sierra

Plegada; LCN = Llanuras de Coahuila y Nuevo Leon; LGM = Laguna de Mayran; LSV = Llanuras y Sierras Volcanicas; PSP =

Pliegues Saltillo-Parras; SLC = Sierras y Llanuras Coahuilenses; SLP = Sierra de la Paila; STR = Sierras Transversales.

a two-volume bilingual compendium of the herpetofauna

of three Mexican states (Chihuahua, Coahuila, and

Sonora), in which they recorded 131 species for

Coahuila. In the same year, Lemos-Espinal (2015)

edited a book on the herpetofauna found in the states

along the Mexico-US border, and recorded 133 species

for Coahuila. Finally, Lemos-Espinal and Smith (2016)

produced a checklist of the Coahuila herpetofauna, in

which they again reported 133 species for the state. The

name usages indicated in the Taxonomic List located at

Amphib. Reptile Conserv.

the Mesoamerican Herpetology website (http://www.

mesoamericanherpetology.com; accessed 8 March 2018)

are followed here.

System for Determining Distributional Status

The system developed by Alvarado-Diaz et al. (2013) for

the herpetofauna of Michoacan is employed to ascertain

the distributional status of members of the herpetofauna

of Coahuila. Mata-Silva et al. (2015), Johnson et al.

October 2019 | Volume 13 | Number 2 | e189

The herpetofauna of Coahuila, Mexico

Fig. 2. Bolsén de Mapimi. Vegetation in the Bolsoén de Mapimi, in the municipality of the same name in the neighboring state of

Durango. Photo by Gabriel Viesca Ramos.

(2015a), Teran-Juarez et al. (2016), Woolrich-Pifia et al.

(2016, 2017), Nevarez-de los Reyes et al. (2016), Cruz-

Saenz et al. (2017), and Gonzalez-Sanchez et al. (2017)

used this system, which consists of the following four

categories: SE = endemic to Coahuila; CE = endemic to

Mexico; NE = not endemic to Mexico; NN = non-native

in Mexico.

Systems for Determining Conservation Status

Assessment of the conservation status of the herpetofauna

of Coahuila, employed the same systems (.e.,

SEMARNAT, IUCN, and EVS) used by Alvarado-Diaz

et al. (2013), Mata-Silva et al. (2015, 2019), Johnson et

al. (2015a,b), Teran-Juarez et al. (2016), Woolrich-Pifia

et al. (2016, 2017), Nevarez-de los Reyes et al. (2016),

Cruz-Saenz et al. (2017), Gonzalez-Sanchez et al. (2017),

and DeSantis et al. (2018). Detailed descriptions of these

three systems appear in earlier papers in this series.

The Mexican Conservation Series

The Mexican Conservation Series (MCS) was initiated

in 2013, with a study of the herpetofauna of Michoacan

(Alvarado-Diaz et al. 2013), as a part of a set of five papers

designated as the Special Mexico Issue published in

Amphibian & Reptile Conservation. The basic format of

the entries in the MCS was established in that paper, 1.e.,

to examine the composition, physiographic distribution,

and conservation status of the herpetofauna of a given

Mexican state or group of states. Two years later, the

MCS was continued with papers on the herpetofauna of

Oaxaca (Mata-Silva et al. 2015) and Chiapas (Johnson

Amphib. Reptile Conserv.

et al. 2015a). In the ensuing year, three entries in the

MCS appeared, those on Tamaulipas (Teran-Juarez et al.

2016), Nayarit (Woolrich-Pifia et al. 2016), and Nuevo

Leon (Nevarez-de los Reyes et al. 2016). Finally, three

entries on Jalisco (Cruz-Saenz et al. 2017), the Mexican

Yucatan Peninsula (Gonzalez-Sanchez et al. 2017), and

Puebla (Woolrich-Pifia et al. 2017) appeared. Thus, this

paper on the herpetofauna of Coahuila is the 10" entry in

this series.

Physiography and Climate

Physiographic Regions

The classification system of physiographic regions

(= subprovinces) developed by INEGI in 2004 was

used to analyze the distribution of the herpetofauna of

Coahuila. This system consists of 10 regions (Fig. 1),

which are briefly described below (see INEGI, http://

www. inegi.org.mx/est/contenidos/proyectos/ce/ce2004/

presentacion.aspx).

Bolsén de Mapimi (BDM). This region, which

encompasses 4,715 km? (3.1% of the state area), is

entirely confined within Mexican territory and runs along

the Sierra Madre Occidental, eventually expanding to the

east in the Mapimi zone. Plains and bajadas dominate

the landscape, although small sierras and lomerios facing

north-south are also found there. The sierras and lomerios

located to the north are composed predominantly of

volcanic rocks and are found associated with faults on

their flanks; to the south limestone is the most abundant

rock. The northern portion of the region is transected by

October 2019 | Volume 13 | Number 2 | e189

Lazcano et al.

Fig. 3. Gran Sierra Plegada. Vegetation in the vicinity of Monterreal in the municipality of Arteaga. Photo by Eli Garcia-Padilla.

the Rio Florido and its effluents, and tributaries of the

Rio Conchos; the southern portion is crossed by the Rio

Nazas. Superficial water sources, however, are scarce.

The Bolson de Mapimi is a flat region at elevations

around 1,200 m, located between Sierra del Diablo, Sierra

Mojada, and irrigation district no. 17. The latter region,

also known as the Comarca Lagunera or La Laguna, used

to be inundated every summer by the waters of the Rio

Nazas until the construction of the Reservoir Francisco

Zarco in the state of Durango. The arid plains of Mapimi

are interrupted by low geomorphological features such

as sand dunes in the northeastern portion. Deep soils of

alluvial or lacustrine origin are well-represented in the

plains. Most of the original vegetation around Torreon,

Matamoros, and San Pedro de las Colonias has been

replaced by agricultural fields. A section of the region

in Laguna del Rey, however, contains microphyll desert

scrub; and a similar habitat is found in the ridges of

Mojada and Montafia del Rey. The middle of this region

contains sand dunes, and the vegetation consists primarily

of Gobernadora/Creosote Bush (Larrea tridentata) and

huizaches (Vachellia [Acacia] spp.), providing minimum

surface cover. The landscapes at La Laguna and El Guaje

are represented by halophytic vegetation, and of less

importance are small areas of grassland and scrubland

where the main plant species is L. tridentata.

Llanuras y Sierras Volcanicas (LSV). This region covers

approximately 14,000 km?(9.2%) of the state of Coahuila,

with elevations ranging between 600 m and 1,200 m. The

largest area of this territory 1s represented by plains or

bajadas; these flat surfaces are more prominent and less

Amphib. Reptile Conserv.

disrupted at Llano de los Ranchos, south of the mountain

ranges that rise along the Rio Bravo/Grande and at

Bolson de los Lipanes to the north of Sierra Mojada.

This region includes small sierras of volcanic rock, such

as Sierra el Mulato, Ocotillo, and Hechicero, located to

the southeast of Ojinaga and the banks of the Rio Bravo.

Small mountains of limestone arise at the southern edge

border with Bolson de Mapimi, as in La Mojada and El

Diablo. Some streams originating in this region feed the

Rio Bravo, and some accumulate water for short periods

of time, but the climate regime 1s that of a desert.

The landscape in this region is dominated by shrubs,

which are generally shorter than two m. Microphyll desert

scrub is found with some variations in its components,

with plains and slopes mostly vegetated by Gobernadora

(L. tridentata), Viscid Acacia/Huizache (Vachellia

vernicosa), Ocotillo (Fouquieria splendens), and mesquite

(Prosopis spp.), there are also other vegetation elements,

such as gatufio/Mimosa (Mimosa spp.), Purple Prickly-

pear (Opuntia macrocentra), and a minor proportion

of huizaches reaching heights of less than 1.5 m. The

components of the lower strata are Cenizo (Leucophyllum

jrutescens), Mariola (Parthenium incanum), Hierba del

Burro/Zinnia (Zinnia acerosa), and Plumed Crinklemat

(Tiquilia greggii). Arborescent yuccas (Yucca spp.) and

Viscid Acacia (V. vernicosa) also are found, reaching

heights of more than four m. This community is widely

distributed in the region, in all the bajadas that feed the

lagoon El Guaje and the western plains.

Laguna de Mayran (LDM). Laguna de Mayran covers

7,804 km? (5.1%) of the state, and is mostly represented

October 2019 | Volume 13 | Number 2 | e189

The herpetofauna of Coahuila, Mexico

No. 1. Anaxyrus speciosus (Girard, 1854). The Texas Toad is distributed from “southeastern New Mexico and western Oklahoma

(USA) south throughout central and West Texas to central Tamaulipas, northern Nuevo Leon, northern and eastern Coahuila, and

northeastern Chihuahua” in Mexico (Frost 2018). This individual came from Allende, in the municipality of Allende. Wilson et al.

(2013b) calculated its EVS as 12, placing it in the upper portion of the medium vulnerability category. Its conservation status has

been considered as Least Concern by IUCN, but this species is not listed by SEMARNAT. Photo by Michael S. Price.

No. 2. Craugastor augusti (Duges, 1879). The Common Barking Frog occurs from “Arizona to Texas in the United States, and

in Mexico from Sonora to Oaxaca, and from Chihuahua, Coahuila, Nuevo Leon, and Tamaulipas to Puebla” (Lemos-Espinal and

Dixon 2013: 42). This individual was found at Cuatrociénegas in the municipality of Cuatrociénegas de Carranza. Wilson et al.

(2013b) ascertained its EVS as 8, placing it in the upper portion of the low vulnerability category. Its conservation status has been

evaluated as Least Concern by IUCN; this species is not listed by SEMARNAT. Photo by Michael S. Price.

Amphib. Reptile Conserv. 36 October 2019 | Volume 13 | Number 2 | e189

Lazcano et al.

No. 3. Lithobates berlandieri (Baird, 1859). The Rio Grande Leopard Frog ranges from “central and western Texas and southern

New Mexico (USA) through eastern Chihuahua to central Veracruz and Hidalgo, Mexico; introduced into the lower Colorado River

and lower Gila River drainages of Sonora and Baja California del Norte, Mexico, and California and Arizona, USA.” (Frost 2018).

This individual was found at El Oso, in the municipality of Cuatrociénegas de Carranza. Wilson et al. (2013b) calculated its EVS

as 7, placing it at the middle portion of the low vulnerability category. Its conservation status has been considered as Least Concern

by IUCN, and as a species of special protection (Pr) by SEMARNAT. Photo by Michael S. Price.

_ el

No. 4. Barisia ciliaris (Smith, 1942). The Sierra Alligator Lizard is a Mexican endemic distributed “along the Sierra Madre Oriental

from Nuevo Leon and southeastern Coahuila southward to at least Guanajuato, and northward along the Sierra Madre Occidental

to extreme southern Chihuahua” (Lemos-Espinal and Dixon 2013: 97). Pictured here is an individual encountered near Monterreal,

in the municipality of Arteaga. Wilson et al. (2013a) determined its EVS to be 15, placing it in the lower portion of the high

vulnerability category. Its conservation status has not been assessed by IUCN, and this species is not listed by SEMARNAT. Photo

by Eli Garcta-Padilla.

Amphib. Reptile Conserv. 37 October 2019 | Volume 13 | Number 2 | e189

The herpetofauna of Coahuila, Mexico

by grassland and some hills found in the municipalities of

Francisco I. Madero, General Cepeda, Parras, San Pedro,

and Viesca. This region includes endorheic terminal

basins of the Nazas and Aguanaval rivers. These rivers

emerge in the Sierra Madre Occidental province, and

flow northward through the province of Mesa del Centro

to the Mayran lagoon. The region is mostly represented

by two bodies of water, Laguna de Mayran, formerly fed

by the Nazas and Viesca rivers, and Laguna de Viesca,

a smaller water body fed by the Aguanaval River. Both

lagoons are located at an elevation of ca. 1,400 m, with

a west-east orientation, and are separated from each

other by a phalanx of the Sierra Madre Oriental. Until

recently, these deposits used to store a significant amount

of water for most of the year, but currently their nearly

level surfaces have turned into desert plains, with saline

areas at the center of the Mayran lagoon and almost

the entire Viesca lagoon. Their disappearance as lakes

and their final passage to a desert regime are the result

of reservoirs and canal systems built on the Nazas and

Aguanaval rivers for irrigation of the Laguna district;

activities that, on the other hand, have significantly

increased productivity in other areas.

Halophytic vegetation is the dominant element of

the landscape, as the soils contain high concentrations

of salts. In fact, roughly from San Pedro de las Colonias

to the boundaries of the Mayran lagoon and throughout

the region, there is no other component than halophytic

vegetation. The area around Mesa Albardienta is

completely devoid of vegetation due to hypersaline

conditions in the soil. Halophytic vegetation in this region

is represented by Saladillo/Salt Bushes (Azriplex spp.),

Seepweeds (Suaeda spp.), and Dropseed (Sporobolus

spp). Besides these types of vegetation, there are small

areas with microphyll desert scrub and rosette scrub in

the Sierrita San Lorenzo.

Sierras y Llanuras Coahuilenses (SLC). This region

covers 43,937 km? (29.0%) of the state. It includes

the municipalities of Abasolo, Frontera, Lamadrid,

Nadadores, Sacramento, and San Buenaventura, as well

as parts of Acufia, Candela, Castafios, Cuatrociénegas,

Escobedo, Monclova, Muzquiz, Ocampo, Progreso, and

Ramos Arizpe, and very small portions of San Juan de

Sabinas and Zaragoza. It consists of folded limestone

mountains ranges, oriented northwest to southeast,

mostly with steep small folds. Most of the mountains lie

between elevations of 1,000 to 2,000 m, although peaks

with elevations over 2,000 m can be found only in Sierra

El Carmen and Sierra de San Antonio. The region has

mostly internal drainage, so its runoff contributions to

the Rio Bravo are minimal.

Vegetation types present in the region are submontane

scrub, chaparral, microphyll desert scrub, and rosette

scrub. The chaparral is generally a dense shrub

community, which is distributed in the transition zone

between the arid scrubland and forests. In this region, it

Amphib. Reptile Conserv.

represents an intermediate layer between the submontane

shrubs and forest, and is also frequently found mixed as

chaparral and desert scrub. It is composed of shrubby

oaks (Quercus spp.) and shrub species in the genera

Cercocarpus and Vauquelinia, among others. Other

shrub components such as sotols (Dasylirion spp.) and

yuccas (Yucca spp.) are also present.

Serranias del Burro (SDB). This region encompasses

13,234 km? (8.7%) of the state, and includes parts of the

municipalities of Acufia, Guerrero, Muzquiz, Sabinas,

Villa Union, and Zaragoza, as well as very small portions

of Juarez, Morelos, and San Juan de Sabinas. The

Serranias del Burro has a normal fault on its northwestern

flank. It is rugged in its central part, which includes a

radial system of narrow valleys, but lies much more

towards the east and southeast, where the sierra becomes

narrow and descends into the hilly zone of Peyotes. A

series of igneous intrusions crosses the Serranias del

Burro from east to west near Villa Acufia and Cerro El

Colorado, with the latter representing the highest peak

of the mountain range, with an elevation of 1,400 m.

The region has few major streams, although it has slopes

toward the Rio Bravo. Soils in the topoform systems

(sierras) constituting this region are mainly represented

by shallow lithosols, while xerosols, phaeozems, and

regosols cover the small systems of hills (lomerios).

Soils on slopes (bajadas) have colluvial or colluvial-

alluvial sources, and the edaphic landscapes in the

intermontane valleys of the Serranias del Burro are very

similar to those in the other regions mentioned above.

The vegetation types found in this region are typical of

the arid zones of Mexico. The hilly zones (lomerios)

that border the mountains of Colorado, del Burro, and

La Babia are covered with rosette scrub. Among the

main components of this vegetation are Lechuguilla

(Agave lechuguilla), Texas Sotol (Dasylirion texanum),

and short yuccas (Yucca spp.). Submontane shrubs on

the Serranias del Burro also ascend the eastern slopes of

the range, with the most distinctive components being

Cenizo/Purple Sage (Leucophyllum frutescens), Tenazas

(Havardia pallens), Hoja Ancha/Tarbush (Flourensia

laurifolia), and Coyotillo/Buckthorn (Karwinskia sp.).

Another vegetation formation of great importance in the

region is chaparral, which grows below the oak forest

on the Sierras del Burro, del Carmen, and La Babia;

this vegetation also grows on the small hills located

immediately to the north of these mountain ranges.

There is also a large number of pine forests in this

region, numerically dominated by Pino Pifionero (Pinus

cembroides). Additionally, there are large extensions of

grassland in the intermontane valleys immediately below

the chaparral.

Finally, in the lower portion of the sierras, adjacent to

the Great Plains of North America, there is Tamaulipecan

thorn scrub. This vegetation is located on the hills

to the south of the region bordering the rosette scrub

October 2019 | Volume 13 | Number 2 | e189

Lazcano et al.

found on the hills of Peyotes. The main components

of this vegetation formation are Palo Verde (Cercidium

texanum), Chaparro } Amargoso/Indian Paintbrush

(Castela texana), Cenizo (Leucophyllum frutescens), and

mesquite trees (Prosopis spp. ).

Sierra de la Paila (SLP). This region includes sierras,

large bolsons with internal drainage, and bajadas. It covers

ca. 19,230 km? (12.7%) of the state. Valle Buenavista

bolson is located in the western portion, bordered to

the west by Sierra de Tlahualilo and to the east by the

highlands of Albardienta, which reach an elevation of

1,800 m. Sierra de La Paila is located to the east, and the

bolsons El Sobaco, El Hundido, San Marcos, and Los

Pinos are located to the north, with the first three at less

than 1,000 m elevation. As in the other regions, the soils

and biodiversity in the Sierra de La Paila is influenced by

climate, which is semiarid at high elevations and very arid

in the grasslands and bolsons. Although topographically

rugged, the sierras have relatively small portions covered

with soils. There are, however, deep soils of primarily

alluvial origin in the lower sections of the bolsons that

have high concentrations of salts. Also, there are sandy

soils of eolic origin that form dunes.

The vegetation communities in this region are mostly

the same as those in Sierras Transversales and Pliegues

Saltillo-Parras. Microphyllous and rosette scrub is

closely associated with the terrain, and dry and semidry

climatic conditions are present in the bajadas and valleys.

Rosette scrub is widely distributed in all sierras such as

La Fragua, La Mesa Albardienta, and La Paila. Chaparral

is a community of shrubs primarily represented by oaks

(Quercus spp.), Chapote/Texas Persimmon (Diospyros

texana), and some elements of rosette scrub, like

yuccas, Sotol, and Lechuguilla. Submontane shrub is

a community composed of shrubs, primarily Chapote

(D. texana), Tenazas (H. pallens), Guajillo (Acacia

berlandieri), and Chaparro Prieto (Acacia amentacea).

These shrubs are also found in La Paila and La Fragua at

the same elevations as chaparral.

Pliegues Saltillo—Parras (PSP). This region covers 9,195

km? (6.1%) of the state of Coahuila. The landscape is

represented by a set of valleys extending from east to

west, situated at an elevation of approximately 1,600 m

and bordered to the north and south by eroded flanks and

valleys. The region also includes the Sierra de Parras,

on which peaks can reach more than 3,000 m, and

includes a succession of truncated large flanks toward

the south. This region in Coahuila includes parts of

the municipalities of Parras, Cepeda, Saltillo, Arteaga,

Ramos Arizpe, Castafios, Candela, and Monclova.

Microphyll desert scrub and rosette scrub are

the dominant vegetation types in this area. Rosette

scrub is distributed on mountain ranges, slopes, and

hills, especially in shallow soils, and alternating with

microphyll scrub in flatter areas, in deep and alluvial soils.

Amphib. Reptile Conserv.

To the north of San Martin de las Varas, rosette scrub

contains some representatives of Pinus cembroides, but

they do not modify the physiognomy of this vegetation

community. Pine forests are also found in the southern

part of the region, on the bajadas of the Sierra El Jabali,

where their density increases with elevation. Between

the scrub and forest, there are two types of natural and

introduced grassland areas. The first area is located south

of General Cepeda, and to the south of Saltillo on the

hills next to Estacion Agua Nueva, and contains grasses

of the genera Bouteloua and Sporobolus. Introduced

grasslands are found to the east of Saltillo, through a

substantial geographic extension of grasses belonging to

the genera Bouteloua and Aristida.

Sierras Transversales (STR). This region extends

throughout the southern section of the state, in the

municipalities of Cuatro Ciénegas, Ocampo, and Sierra

Mojada, encompassing a surface area of 14,077 km? (9.3%

of the state area). Over half of this region consists of sierras

with shallow light-colored soils. The main vegetation

is represented by rosette scrub and microphyll desert

scrubland. Rosette scrub vegetation is distributed on all

the mountains, slopes, and small hills located at elevations

between 2,000 and 2,400 m, with large portions of this

vegetation located on the sierras El Numero, Candelaria,

Parras, and other smaller sierras. The main components of

this vegetation are Huizache (Acacia farnesiana), Chapotes

(D. texana), Texas Sotol (D. texanum), Lechuguilla (A.

lechuguilla), and Gatufio (Acacia roemeriana). In the

lower parts of the sierras Playa Madero and del Laurel,

the same type of vegetation is also found, where the

numerically dominant Yucca thompsoniana gives the

appearance of an Izotal (Yucca tree forest), in addition to

Sotol, Lechuguilla, Chaparro Prieto (Acacia rigidula), and

Fresno (Fraxinus greggii), among others. This association

also is present on the bajadas of the sierras.

The abundance of microphyll shrubs is noteworthy

on flat areas, especially in the southeastern part of the

state, with thorny elements present, such as mesquites

(Prosopis spp.) and huizaches (Acacia spp.), as well as

Gobermadora (L. tridentata) and Hojasén (Fluorencia

cernua). Additionally, relatively small areas of grassland

are found on the sierras of GOmez Farias and Jabali,

with species of the genera Bouteloua, Muhlenbergia,

Andropogon, and Aristida. The southeastern section of

the state also has flat areas with halophytic vegetation

on saline soils, with species of saladillo (Suwaeda spp.)

and saltbushes (Atriplex spp.). On the other hand,

chaparral, pine-oak forest, pine forest, and small areas

with submontane scrub are found in the less arid sierras,

such as the Sierras de Jimulco, Parras, and Jabali. Among

components of the chaparral are small oaks (Quercus

spp.) and Pino Pifionero (P. cembroides).

Gran Sierra Plegada (GSP). This region covers 2,178

km? (1.4%) of the state. It includes a major portion of the

October 2019 | Volume 13 | Number 2 | e189

The herpetofauna of Coahuila, Mexico

Table 1. Monthly minimum, mean (in parentheses), maximum, and annual temperature data (in °C) for the physiographic regions of Coahuila, Mexico. The locality and

elevation for each region are: Bolson de Mapimi

Laboratorio del Desierto, Tlahualilo, Durango (1,160 m); Llanuras y Sierras Volcanicas

Laguna de Mayran—Viesca (1,100 m); Sierras y Llanuras Coahuilenses—San Francisco Nadadores (500 m); Serrania del Burro—Agua Nueva (370 m); Sierra La Paila

Sierra Mojada (1,256 m);

Hipolito (1,150 m); Pliegues Saltillo-Parras—General Cepeda (1,400 m); Sierras Transversales—La Ventura (1,867 m); Gran Sierra Plegada—San Antonio de las Alazanas

(2,300 m); and Llanuras de Coahuila y Nuevo Le6n—Presa Venustiano Carranza (272 m). Data from: http://www. smn1.conagua.gob.mx/climatologia/normales/estacion/

EstacionesClimatologicas.kmz; accessed 11 November 2017.

Physiographic Region Jan Feb Mar Apr May

Bolson de Mapimi 3.0 5.0 8.2 12.4 16.5

(11.7) (14.0) (17.4) (21.5) (25.3)

20.3 23.0 26.7 30.6 34.1

Llanuras y Sierras 4.3 De 8.2 12.0 15.0

Volcanicas (10.6) (12.2) (15.6) (19.5) = (22.6)

16.8 18.9 23.0 27.0 30.3

Laguna de Mayran 40 6.2 9.0 13.0 16.7

(14.0) (16.4) (19.6) (23.3) (26.7)

24.0 26.7 30.3 33.6 36.6

Sierras y Llanuras 2.8 47 7.5 1 Ui 16.3

Coahuilenses (11.0) (14.0) (17.4) (21.5) ~~ (25.7)

19.3 233 212 31.6 35.2

Serrania del Burro 46 5.0 7.5 123: 15.7

(11.7) (12.7) (15.8) (20.7) (23.8)

18.9 20.5 24.0 29,2, 31.9

Sierra La Paila 9.6 10.7 12.8 14.5 17.6

(15.0) (16.0) (17.9) (20.2) (23.0)

20.3 21:3 23.1 25:9 28.3

Pliegues Saltillo-Parras 5.3 6.5 9.4 12.9 15.8

(12.8) (14.4) (17.9) (21.2) (24.0)

20.2 223 263 29.5 32.1

Sierras Transversales 19 em a3 Le 10.9

(11.7) (13.1) = (16.0) (18.5) (21.6)

21.5 23-1 26.8 29.4 32.3

Gran Sierra Plegada 42 5.0 6.2 7.8 9.0

(12.2) (13.2) (14.6) (16.4) (17.5)

20.2 21.3 22.9 25. 25.9

Llanuras de Coahuila y 49 7.0 10.7 15.1 19.0

Nuevo Leon (11.9) (14.5) (18.5) (22.8) ~—_ (26.0)

18.9 22-0 26.2 30.5 33.1

municipality of Arteaga (95%) and a very small fraction of

Saltillo. This region begins east of Saltillo, Coahuila, but also

includes sections in Nuevo Leon, Tamaulipas, and San Luis

Potosi, and is dominated by folded layers of limestone. A

great reverse geological fault lies on the eastern edges of the

Gran Sierra Plegada, while smaller ones extend relatively

parallel to it and its structural axes. The elevational range in

the region is between 2,000 and 3,750 m.

The topography is predominantly mountainous, but

also contains plateaus and valleys. This region 1s located

somewhat parallel to the Gulf of Mexico and represents an

orographic barrier that favors the deposition of moisture

on the eastern slopes, which prevents the westward

movement of moist winds. Heavy rainfall has led to the

dissolution of limestone rocks in the area, resulting in

a karstic environment. These processes have led to the

formation of vast cavern systems and springs at the foot

of the mountains. A broad elevational gradient is present

in this area. The soils are dominated by lithosols, which

are associated with rendzinas and calcaric regosols.

Amphib. Reptile Conserv.

Jun Jul Aug Set Oct Nov Dec

19.1 19.2 18.6 16.7 12.7 TS 33

(27.2) (26.5) (25.8) (23.8) (20.6) (15.8) (12.0)

35.3 33,9 33.0 31.0 28.5 24.2 20.6

16.7 16.4 16.0 14.1 11.0 7.0 49

(24.3) (23:5) (22:9) (20.8) (17.9) (14.1) (11.1)

31.8 30.5 29-7, 215 24.9 971 Uh | 17.4

19.5 20.2 195 18.0 13.4 7.7 46

(28.3) (28.1) (27.6) (26.0) (22.4) (17.8) (14.4)

37.1 36.0 35.7 33.9 31.5 27.8 24.2

19:2 19,2 20%) 17.0 12:5 Tee 41

(28.0) (27.8) (28.3) (24.8) (20.7) (15.8) (12.1)

36.7 36.3 36.5 Sap 28.8 24.3 20.2

18.4 18.8 18.9 16.3 a Tad 44

(25.8) (25.8) (25.1) (22.8) (18.6) (14.5) (10.6)

33.3 32.8 33 292 25:1 212 16.7

19.5 19.5 19.4 17.6 15.9 13.1 11.1

(25.1) (25.2) (24.8) (22.6) (21.0) (18.4) (16.1)

30.8 30.9 30.2 27.6 26.1 23.6 211

17.5 17.4 16.9 15.0 12.2 8.4 6.2

(25.0) (24.6) (24.0) (21.8) (19.5) (15.9) (13.5)

32.5 31.8 31.0 28.6 26.7 23.4 20.8

(bees) 13.3 12.6 12.2 8.8 5.6 2.1

(22.5) (22.3) (21.7) (21.5) (19.4) (15.5) (12.0)

32.6 31.3 30.8 30.8 29.9 255 21.9

9:5 9.6 9.1 8.6 2 5.6 46

(17.5) (17.3) (16.9) (16.6) (15.6) (14.2) (12.7)

25.5 24.9 24.4 24.6 23.7 22:9 20.7

21.7 22.6 po) 20.3 15:7 9.8 5.6

(28.7) (29.5) (29.3) (26.7) (22.3) (16.8) (12.8)

35:7, 36.5 36.1 33.1 28.9 23.8 19.9

Calcic and haplic xerosols are also found within the

region.

In general, two fundamental forms of plant landscapes

are present in the region: forests and scrublands. Pines

dominate the forested area, and desert rosette scrub,

piedmont scrub, and chaparral dominate the rest of the

region. Other types of vegetation in the Gran Sierra

Plegada occur as small patches of grassland, halophytic

vegetation, or alpine prairie, but they have minimal

influence in shaping the overall landscape.

Llanuras de Coahuila y Nuevo Leon (LCN). This region

encompasses 25,666 km? (16.9% of the state area),

including the municipalities of Hidalgo, Nava, Piedras

Negras, and Jiménez, and parts of Guerrero, Villa Union,

Morelos, Allende, Progreso, Escobedo, Sabinas, San

Juan de Sabinas, Nueva Rosita, Muzquiz, Zaragoza,

and Acufia. The region is characterized by the presence

of plains interrupted with scattered low hills that are

composed of conglomerates, at elevations ranging from

40 October 2019 | Volume 13 | Number 2 | e189

Annual

11.8

(20.1)

28.4

10.9

(17.9)

24.9

12.7

(22.1)

31.5

11.9

(20.6)

29.3

11.8

(19.0)

26.2

15.1

(20.4)

25.8

12.0

(19.6)

27.1

8.0

(18.0)

28.0

(15.4)

23.5

14.6

(21.7)

28.7

Lazcano et al.

Fig. 4. Llanuras de Coahuila y Nuevo Leon. Tamaulipas thorn

scrub in the municipality of Allende. Photo by Manuel Nevarez

de los Reyes.

Fig. 6. Llanuras y Sierras Volcdnicas. Vegetation near Heér-

cules, in the municipality of Sierra Mojada. Photo by Daniel

Solorio Estrada.

75 to about 500 m. One of the most extensive plains

extends from Anahuac, Nuevo Leon, to Nueva Rosita,

Coahuila, at an average elevation of 500 m.

Tamaulipan thornscrub and mesquites (Prosopis spp.)

are the most characteristic vegetation types in this region.

Tamaulipan thornscrub is distributed at elevations from 80

to 340 m, witha physiognomy of thornscrub in areas of low

relief and of semi-thorn scrubland on the lower sections

of areas with higher relief. Large patches of Cenizo (L.

frutescens) are present in some areas, indicative of a high

degree of disturbance to the native scrub vegetation, as

this species numerically dominates the sympatric native

species that are found in low frequency and are small in

size. Mesquites dominate at elevations from 75 to 400 m.

Piedmont scrub or Tamaulipan thornscrub predominate

in some middle sections, and a prevalence of halophytic

elements is present in the lower areas. Some deciduous

thornscrub and deciduous hardwood forests are found in

the region as well, and oak and pine-oak forests occur at

higher elevations. Halophytic vegetation is found within

small areas of the plains and valleys, where high salt

concentrations are present in the soils. Natural grassland

Amphib. Reptile Conserv.

Fig. 5. Laguna de Mayrdn. Vegetation on Cerro de la Virgen,

in the municipality of Parras. Photo by José Flores Ventura.

SE UAW Dee eg tte ee ae

Fig. 7. Pliegues Saltillo-Parras. Vegetation near Parras de la

Fuente, in the municipality of Parras. Photo by Manuel Nevdrez

de los Reyes.

occurs in some areas of the plains at elevations from 135

to 290 m. The introduced grasses on the plains and valleys

are composed primarily of Buffelgrass (Pennisetum

ciliare), which 1s distributed at elevations from 190 to

270 m and covers small hilly areas and alluvial plains.

Climate

Temperature. The minimum, mean, and maximum

temperatures for one locality in each of the 10

physiographic regions in Coahuila are shown in Table 1.

The elevations for these 10 regions range from 272 m in

the Llanuras de Coahuila y Nuevo Leon to 2,300 m in the

Gran Sierra Plegada.

The mean annual temperature (MAT) of these regions

ranges from a low of 15.4 °C, in the Gran Sierra Plegada

at 2,300 m in the southeastern portion of the state, to a high

of 22.1 °C, in the Laguna de Mayran at 1,100 m in the

southern portion of the state. The MAT of these regions in

Coahulia are unusual tn that they do not gradually decrease

with increasing elevation. The MAT lie below 20 °C in

four additional regions, including the Llanuras y Sierras

October 2019 | Volume 13 | Number 2 | e189

The herpetofauna of Coahuila, Mexico

Table 2. Monthly and annual precipitation data (in mm) for the physiographic regions of Coahuila, Mexico. The locality and elevation

for each region are: Bolson de Mapimi—Laboratorio del Desierto, Tlahualilo, Durango (1,160 m); Llanuras y Sierras Volcanicas—

Sierra Mojada (1,256 m); Laguna de Mayran—Viesca (1,100 m); Sierras y Llanuras Coahuilenses—San Francisco Nadadores (500

m); Serrania del Burro—A gua Nueva (370 m); Sierra La Paila—Hipolito (1,150 m); Pliegues Saltillo-Parras—General Cepeda (1,400

m); Sierras Transversales—La Ventura (1,867 m); Gran Sierra Plegada—San Antonio de las Alazanas (2,300 m); Sierras y Llanuras

Occidentales—Carbonera, Galeana, Nuevo Leon (2,035 m); and Llanuras de Coahuila y Nuevo Leon—Presa Venustiano Carranza

(272 m). The shaded area indicates the months of the rainy season. Data from: http://www. smn1.conagua.gob.mx/climatologia/

normales/estacion/EstacionesClimatologicas.kmz; accessed 11 November 2017.

PBoiséndeMapimi | 95 [40 | 42 | 89 | 152 | 362 | 466 | 548] a4 [208] 91 | 69 | 2586 |

[sierataPaila ——-| 90 [91 | 8 | 82 | 168 | aso [291 | 23 | an7 | 27 | se | 86 | i681 |

Volcanicas at 1,256 m (17.9 °C), the Serranias del Burro

at 370 m (19.0 °C), the Pliegues Saltillo-Parras at 1,400 m

(19.6 °C), and the Sierra Transversales at 1,867 m (18.0°C).

In contrast, the MAT in the remaining five regions lie

above 20 °C, including the Bolson de Mapimi at 1,160 m

(20.1 °C), the Laguna de Mayran at 1,100 m (22.1°C), the

Sierra de Paila at 1,150 m (20.4 °C), the Sierras y Llanuras

Coahuilenses at 500 m (20.6 °C), and the Llanuras de

Coahuila y Nuevo Leon at 272 m (21.7 °C).

The minimum annual temperature ranges from 7.2

°C in the Gran Sierra Plegada to 15.1 °C in the Sierra

La Paila (Table 1). The maximum annual temperature

varies from 23.5 °C in the Gran Sierra Plegada to 31.5

°C in the Laguna de Mayran. The minimum annual

temperature is 10.7—20.0 °C lower than the maximum

annual temperature among the 10 physiographic regions

of the state (Table 1). Mean monthly temperatures peak

at some point from May to August, usually June, and

reach a low point sometime during December or January,

usually January (Table 1).

Precipitation. As expected, precipitation in Coahuila is

generally highest during the rainy season from June to

October, and lowest from November to May, during the

dry season. The data in Table 2 indicate that 56.8—-77.6%

(x = 69.4%) of the rainfall occurs during the rainy season.

Table 3. Composition of the native and non-native herpetofauna

of Coahuila, Mexico.

The month with the greatest amount of precipitation,

depending on the location, is June, July, August, or

September, usually August (Table 2). The month with

the least amount of precipitation, again depending on

the location, is December, February, or March, usually

March (Table 2). The annual precipitation ranges from

96.9 mm in the Serranias del Burro to 384.3 mm in the

Llanuras de Coahuila y Nuevo Leon.

Composition of the Herpetofauna

Families

The herpetofaunal species of Coahuila are placed

in 28 families, including seven for anurans, two for

salamanders, 15 for squamates (one of which contains

only a single non-native species), and four for turtles

(Table 3). The seven anuran families comprise 63.6% of

the 11 families with representatives in Mexico. The two

salamander families constitute 50.0% of the four families

represented in the country. The fifteen squamate families

make up 48.4% of the 31 Mexican families containing

native species. Finally, the four Coahuilan turtle families

encompass 40.0% of the 10 families in Mexico. The total

of 28 families includes 47.5% of the 59 herpetofaunal

families that are represented in this country (Johnson et

al. 2017). There are no caecilian or crocodylian families

with representatives in Coahuila.

Orders Families Genera Species Genera

Anura 7 13 20

Caudata 2 3 4 Sixty-six herpetofaunal genera are represented

Subtotals 9 16 74 in Coahuila, including 13 for anurans, three for

Squamata 15 44 106 salamanders, 44 for squamates, and six for turtles (Table

Testudines 4 6 13 3). The 13 anuran genera constitute 35.1% of the 37

Subtotals 19 50 119 with representatives in Mexico. The three salamander

Totals 28 66 143 genera make up 15.8% of the 19 found in Mexico. Of the

Amphib. Reptile Conserv.

October 2019 | Volume 13 | Number 2 | e189

Lazcano et al.

rm ¥ 7. valu. # “et fy ae ;

No. 5. Gerrhonotus infernalis Baird, 1859. The Texas Alligator Lizard ranges “from central Texas westward to the area of Big Bend,

in the United States, and in Mexico east of the Sierra Madre Oriental to southern San Luis Potosi and perhaps extreme southeastern

Durango” (Lemos- Espinal and Dixon 2013: 98). This individual was found in Sierra La Concordia, in the municipality of General

Cepeda. Wilson et al. (2013a) calculated its EVS as 13, placing it at the upper end of the medium vulnerability category. Its

conservation status has been gauged as Least Concern by IUCN, and this species is not listed by SEMARNAT. Photo by Michael

S. Price.

a. lal _ . S

= A

es - >

b Soap --'* . ¢ : a &

* ; ; ,

7 J : «fT - . * p- : . 7 ~

4, - * «

w eb . -— : : oe ba

No. 6. Gerrhonotus lugoi McCoy, 1970. Lugo’s Alligator Lizard is a Mexican endemic species restricted to the Cuatro Ciénegas

region (Lemos-Espinal et al. 2015). This individual was found at Cuatrociénegas, in the municipality of Cuatrociénegas de Carranza.

Wilson et al. (2013a) judged its EVS as 17, placing it in the middle portion of the high vulnerability category. Its conservation status

is evaluated as Least Concern by IUCN, and as threatened (A) by SEMARNAT. Photo by Michael S. Price.

Amphib. Reptile Conserv. 43 October 2019 | Volume 13 | Number 2 | e189

The herpetofauna of Coahuila, Mexico

a\, & vs Pa Se

2 See Ps

No. 7. Gerrhonotus mccoyi Garcia-Vazquez, Contreras-Arquieta, Trujano-O

ega, and Nieto-Montes de Oca, 2018. McCoy’s

Alligator Lizard is a Mexican endemic species restricted to the Cuatro Ciénegas region (Garcia-V a4zquez et al. 2018). This individual,

a male paratype of the species, was encountered at Pozas Azules, Rancho Pronatura in the municipality of Cuatro Ciénegas. The EVS

of this species can be calculated as 6+8+3=17, placing it in the middle portion of the high vulnerability category. Its conservation

status has not been evaluated by IUCN, and this species is not listed by SEMARNAT. Photo by Uri Garcta-Vdzque:.

7-3 4 ae ~~? :

and Solis in extreme western Coahuila” (Lemos-Espinal et al. 2015: 173). This individual was encountered at ca. 8 km SW from the

locality of San Antonio del Coyote, in the municipality of Matamoros. Wilson et al. (2013a) determined its EVS as 16, placing it in

the middle portion of the high vulnerability category. Its conservation status has been evaluated as Endangered by IUCN, but it has

not been listed by SEMARNAT. Photo by Marco Antonio Bazan-Tellez.

Amphib. Reptile Conserv. 44 October 2019 | Volume 13 | Number 2 | e189

Lazcano et al.

oe a aa

: * pita fn

= Pane

by Daniel Garza-

Fig. 8. Serranias del Burro. Panoramic view of the Serranias del Burro, in the municipality of Zaragoza. Photo

Tobon.

138 genera of squamates in Mexico, the 44 represented Ambystoma velasci. Although Lemos-Espinal and Smith

in Coahuila amount to 31.9%. The six turtle genera (2016) reported A. mavortium in Coahuila, we consider

comprise 33.3% of the 18 genera with representatives in populations of Ambystoma in southeastern Coahuila to

Mexico. The total of 66 genera encompasses 30.6% of — belong to the same species that occurs in nearby Nuevo

the 216 found in Mexico (Johnson et al. 2017). Leon, which was identified as A. velasci by Nevarez-de

los Reyes et al. (2016).

Species

Gerrhonotus mccoyi. Recently, Garcia-Vazquez et al.

The herpetofauna of Coahuila represents 143 species, (2018) described this species from the Cuatro Ciénegas

including 20 anurans, four salamanders, 106 squamates, — Basin in the Sierras y Llanuras Coahuilenses region.

and 13 turtles (Table 3). The 20 anuran species comprise

8.1% of the 247 distributed in Mexico. The four Gerrhonotus parvus. Until recently, this species was

salamander species constitute 2.6% of the 151 found in —_ considered endemic to Nuevo Leon (Lemos-Espinal et

Mexico. The 106 squamate species make up 12.3% ofthe al. 2016). Banda-Leal et al. (2018), however, reported

863 located in Mexico. The 13 turtle species amount to it from Sierra de Zapalinamé, Coahuila. Therefore,

25.5% of the 51 species occurring in Mexico. The total — this species presumably has a continuous distribution

of 143 species comprises 10.8% of the 1,318 species throughout the Gran Sierra Plegada region between

making up the Mexican herpetofauna (Johnson, unpub.). —' the type locality (Galeana, Nuevo Leon) and Sierra

Zapalinameé, Coahuila.

Comments on the Species List

Sceloporus bimaculosus. A former subspecies of S.

Some comments on our list of recognized species are = magister (elevated by Schulte et al. 2006) was returned

necessary, especially as compared to that in Lemos- to the synonymy of S. magister by Leaché and Mulcahy

Espinal and Smith (2016), as follows: (2007). Curiously, a few sources since then (including

Lemos-Espinal and Smith’s (2016) most recent checklist

Eleutherodactylus marnockii. Although Lemos-Espinal on the herpetofauna of Coahuila and the Reptile Database

and Smith (2016) listed this frog as aresident of Coahuila, — website) referenced the former publication and continued

they provided no evidence based on voucher specimens. _ to recognize S. bimaculosus as a full species despite a short

In addition, neither Dodd (2013) nor Frost (2018) list this discussion of that issue by Wilson et al. (2015). In any event,

species as occurring 1n Mexico. Thus, we do not include _at this point, we herein recognize S. magister as the species

this species in our analysis.

Amphib. Reptile Conserv. 45 October 2019 | Volume 13 | Number 2 | e189

The herpetofauna of Coahuila, Mexico

of the S. magister species group occurring in Coahuila.

Sceloporus cowlesi. Lemos-Espinal and Smith (2016) in

their recent checklist reported Sceloporus consobrinus

for the Coahuila herpetofauna. In a study of the molecular

phylogenetics of the Sceloporus undulatus species

group, however, Leaché (2009) restricted the distribution

of S. consobrinus to the United States and indicated the

member of the wndulatus group found within Nuevo

Leon to be S. cowlesi, which is the name we use here for

the Coahuilan populations.

Sceloporus gadsdeni. Diaz-Cardenas et al. (2017)

recently described this species from Sierra de San

Lorenzo, near Torreon, in the Bolson de Mapimi region.

Sceloporus ornatus. Herein we regard Sceloporus

oberon as a synonym of Sceloporus ornatus, based on

Martinez-Méndez and Méndez-de la Cruz (2007).

Sceloporus marmoratus. Unlike Lemos-Espinal and

Smith (2016), we do not list S. variabilis as occurring in

Coahuila, but based on Mendoza-Quiyano et al. (1998)

we do recognize S. marmoratus as occurring in the

Llanuras de Coahuila y Nuevo Leon region of Coahuila,

instead of S. variabilis.

Lampropeltis annulata. Lampropeltis annulata was

considered an evolutionary species separate from L.

triangulum by Ruane et al. (2014), who elevated a

number of subspecies of L. triangulum to full species

and synonymized others. Subspecies synonymized with

L. annulata included L. t. dixoni.

Lampropeltis gentilis. The first record for this species

from Mexico and Coahuila was reported by Baeza-Tarin

et al. (2018a).

Lampropeltis splendida. Lampropeltis splendida was

elevated to a full species separate from L. getula by

Pyron and Burbrink (2009).

Salvadora deserticola. Nevarez de los Reyes et al. (2018)

first reported this species from Coahuila.

Tantilla cucullata. Baeza-Tarin et al. (2018b) first

reported this species for Coahuila.

Trimorphodon vilkinsonii. Baeza-Tarin et al. (2018c)

first reported this species for Coahuila.

Crotalus ornatus. Anderson and Greenbaum (2012)

resurrected Crotalus ornatus from the synonymy of

C. molossus. Crotalus ornatus is found in most parts

of Coahuila, whereas C. molossus is restricted to the

extreme southern parts of the state.

Amphib. Reptile Conserv.

Apalone atra. Apalone atra has been either regarded as a

subspecies of A. spinifera or as a species endemic to the

Valley of Cuatro Ciénegas, where A. spinifera has gained

access to some areas through irrigation channels, thereby

allowing some genetic introgression to take place, and

driving A. atra to a level of being critically susceptible

to extinction. Smith and Smith (1979) had already

considered A. atra to be extinct. Recently, however, pure

individuals of A. atra have been found, as well as hybrids

between the two species (Cerda-Ardura et al. 2008).

See Wilson and Johnson (2010) for a discussion on this

issue. Until updated data indicate otherwise, we regard

A. atra as having viable populations in the Valley of

Cuatro Ciénegas, highlighting its need for conservation

assessment and action.

Patterns of Physiographic Distribution

Herein 10 physiographic regions are recognized in

Coahuila (Fig. 1), and the occurrence of the members of

the herpetofauna among these 10 regions are shown in

Table 4 and summarized in Table 5.

The total number of species in each region ranges

from a low of 38 in the Laguna de Mayran to a high

of 91 in the Sierras y Llanuras Coahuilenses (Table

5). The number of species in each of the other regions

is as follows, in ascending order: 40 (Sierra de la

Paila); 44 (Llanuras y Sierras Volcanicas); 45 (Bolson

de Mapimi); 45 (Serranias del Burro); 47 (Sierra

Transversales); 49 (Pliegues Saltillo Parras); 51 (Gran

Sierra Plegada); and 53 (Llanuras de Coahuila y Nuevo

Leon). The lowest value of 38 in the Laguna de Mayran

is 41.8% of the highest value of 91 in the Sierras y

Llanuras Coahuilenses. The latter region is the largest

in the state, but the former region is not the smallest (the

smallest region is the Gran Sierra Plegada).

As expected, the largest absolute and relative numbers

of the component herpetofaunal groups are found in the

Sierras y Llanuras Coahuilenses, including 14 of 24

species of amphibians (58.3%), 68 of 106 species of

squamates (64.2%), and nine of 13 species of turtles

(69.2%).

Members of the Coahuilan herpetofauna inhabit from

one to all 10 of the 10 physiographic regions, as follows:

one (66 of 143 species: 46.2%); two (25; 17.5%); three

(11; 7.7%); four (five; 3.5%); five (0; 0%); six (0; 0%);

seven (two; 1.4%); eight (five; 3.5%); nine (11; 7.7%);

and 10 (18; 12.6%). The most broadly distributed species

(occupying all 10 regions) are the anurans Anaxyrus

debilis, A. punctatus, Lithobates berlandieri, Scaphiopus

couchii, and Spea multiplicata;, the lizards Crotaphytus

collaris, Coleonyx brevis, Hemidactylus turcicus (non-

native), Phrynosoma cornutum, Sceloporus grammicus,

S. poinsetti, and Aspidoscelis gularis, and the snakes

Lampropeltis splendida, Masticophis flagellum,

Pantherophis emoryi, Pituophis catenifer, Rhinocheilus

lecontei, and Thamnophis marcianus. Given that Coahuila

October 2019 | Volume 13 | Number 2 | e189

Lazcano et al.

Table 4. Distribution of the amphibians, squamates, and turtles of Coahuila, Mexico, by physiographic region. Abbreviations are as

follows: BDM = Bolson de Mapimi; LSV = Llanuras y Sierras Volcanicas; LDM = Laguna de Mayran; SLC = Sierras y Llanuras

Coahuilenses; SDB = Serranias del Burro; SLP = Sierra de la Paila; PSP = Pliegues Saltillo Parras; STR = Sierras Transversales;

GSP = Gran Sierra Plegada; and LCN = Llanuras de Coahuila y Nuevo Leon. * = species endemic to Mexico; ** = species endemic

to Coahuila; and *** = non-native species. See text for detailed descriptions of these regions.

Physiographic Regions of Coahuila Number

Taxa of Regions

| Bufonidae(7speciesy | | EE

ee ee a a

Anasymis debits a oe eae ae ee io |

ce a i Co

ee a a (i ee QR a a

| Anaxyruswoodhousii_ | + OL + TT

| Rhinellahorribilis | Et TE

| Craugastoridae(I species) | | EE

| Craugastoraugusti | Tt

| Eleutherodactylidae(3speciesy) | | | TT

| Eleutherodactylus cystignathoides_ | |_| ST ST CE TE

| Eleutherodactylus gutilatus | | TE + TOE CE vt TT

| Eleutherodactyluslongipes* | | ST TE Et Tt |

| Hylidae(4speciesy | |

| Acris blanchardi TT tT

| Dryophytesarenicolor | TE Et TCE

| Rheohylamiotympanum® | LE

| Smiliscabaudinii | Et

| Microhylidae(1 species) | | TE

| Gastrophryne olivacea | + | + | + [| + | + P+] + To + | +t TT

| Ranidae(2speciesy) | |

| Lithobates catesbeianus*** | + [Tt Tt 8

| Scaphiopodidae(2 species) | | | EE

| Caudata(4speciesy |

| Ambystomatidae(I species) | | | TE EE

| Ambystomavelasci* | TE Et

| Plethodontidae(3speciesy | | TE EE

| Aquiloewryceagaleanae* | TT tT

| Aquiloeurycea scandens* | TT tT

| Chiropterotritonpriscus* | TE tT

| Squamata(106 species) | | TE

| Anguidae(Sspeciesy | | TE

Be ee i a a ee ee ee eee ee

| Gerrhonotusinfornalis, | ee eee +t ee

| Gerrhonostugoi** | Tt

| Gerrhonotusmecoyi** | Tt TE

| Gerrhonotusparvus* | TE

| Crotaphytidae(4 species) | |

| Crotaphytusantiquis** | Tt

| Crotaphytusreticulatus | LE

| Gambeliawistizeni | Lt TE

| Eublepharidae(2species) | | | CE

| Coleonyxreticulas | Tt TE

| Gekkonidae(I species) | | EC CC CE

Amphib. Reptile Conserv. 47 October 2019 | Volume 13 | Number 2 | e189

The herpetofauna of Coahuila, Mexico

Table 4 (continued). Distribution of the amphibians, squamates, and turtles of Coahuila, Mexico, by physiographic region.

Abbreviations are as follows: BDM = Bolson de Mapimi; LSV = Llanuras y Sierras Volcanicas; LDM = Laguna de Mayran; SLC =

Sierras y Llanuras Coahuilenses; SDB = Serranias del Burro; SLP = Sierra de la Paila; PSP = Pliegues Saltillo Parras; STR = Sierras

Transversales; GSP = Gran Sierra Plegada; and LCN = Llanuras de Coahuila y Nuevo Leon. * = species endemic to Mexico; ** =

Species endemic to Coahuila; and *** = non-native species. See text for detailed descriptions of these regions.

Physiographic Regions of Coahuila Number

Taxa of Regions

[Phrynosomatidae @9 species) |_| _ =r

ee ee SS

| Holbrookia approximans* |_| + | + | + | + [+ }]+] +] + | | 9 |

| Holbrookialacerata | | | CCT CCT CT Cs Tt

| Phrynosoma modestum ss |_+ | + | + | + | + [+] +] +] + | | 9 |

| Sceloporuscautus* | tT TT CT CT

| Sceloporuscouchii* | | dT dE + TUT + T+ T+} TUT

[| Sceloporuscowlesi | + | + | + | + | + f+ t+] +] + | [| 9 |

| Sceloporuscyanogenys |_| | dT + | UT CT

| Sceloporuscyanosticius* |_| | CT CT CT C+ T+ TUT

| Sceloporusgadsdeni** | + | LT dT CT CT

| Sceloporusgoldmani* | | | CT CCT CT CT Tt

| Sceloporus grammicus | + | + | + | + | + |+ | +] +] + | + | 0

[| Sceloporusmagister | | *# [| TE + | oT + TUT CUT CCU

| Sceloporusmarmoraus {|_| | | CCE CT CT

[| Sceloporusmerriami | tT dT dE + TOUT + | + Ts |

| Sceloporusminor* | tT TT CT CT

| Sceloporusolivaceus | | | dT + T+ TUT CU

| Sceloporusornatus* | TT TE CT CT CT} TU

| Sceloporusparvus* | tT TE TO

| Sceloporuspoinsetii | + | + | + | + | + P+ t+ P+ | + | + |

| Sceloporussamcolemani* |_| | dT CT CT CT Tt

A EO ee

Uma exsul**

a a | a! a a a a (ar ay

| Urosaurusornaus | + TL dE +hT + TUT

| Uta stansburiana | +} TE + | +! T+ |] T+ T+ T+! T+! TUT 8

| Plestiodondicei* | | CT CT CT CT

| Plestiodon obsolems | + | + | + | + | + | +> +] +] t+] 9 |

| Plestiodon tetragrammus {|_| | dL + T+ TUT CU

| Sphenomorphidae(3 species) {|__| | S| CT CT CT

| Scincellakikaapoa** | | dT Cd + TUT CT

| Scincellalateralis | | CT CE + TUT CT

| Scincellasivicola* | | dT CL CCT CT Tt

| Teiidae(4speciesy | | CT CT CCT CC

| Aspidoscelis gularis | + | + | + | + | + |+ | +] +] + | + | 0 |

| Aspidoscelis inornata | + | + | + | + | + |+ t+] +] + | | 9

| Aspidoscelis marmorata | _ + | + | + | + | + [+ t]+t] +t] | | 8 |

| Aspidoscelis tesselaa | | + | | + | tT TC

| Xantusiaextorris* | ET

| Colubridae(29speciesy | | CE | CE CE

| Arizonaelegans | +} Tc *® | +! T+ T+ t+ P+] oT hut +h TL 8

| Bogertophis subocularis | + | + | + | + | + [+ ]+] | | | 7 |

| Coluber conswrictor | dT dT CT + TUT CT CO

NPENTNP Re fee _wepTy

Amphib. Reptile Conserv. 48 October 2019 | Volume 13 | Number 2 | e189

Lazcano et al.

Table 4 (continued). Distribution of the amphibians, squamates, and turtles of Coahuila, Mexico, by physiographic region.

Abbreviations are as follows: BDM = Bolson de Mapimi; LSV = Llanuras y Sierras Volcanicas; LDM = Laguna de Mayran; SLC =

Sierras y Llanuras Coahuilenses; SDB = Serranias del Burro; SLP = Sierra de la Paila; PSP = Pliegues Saltillo Parras; STR = Sierras

Transversales; GSP = Gran Sierra Plegada; and LCN = Llanuras de Coahuila y Nuevo Leon. * = species endemic to Mexico; ** =

Species endemic to Coahuila; and *** = non-native species. See text for detailed descriptions of these regions.

Physiographic Regions of Coahuila Number

Taxa of Regions

LCN Occupied

LDM T

Lampropeltis annulata

Lampropeltis gentilis

Lampropeltis leonis*

Lampropeltis splendida

—

Masticophis flagellum

Masticophis schotti

Masticophis taeniatus

Opheodrys aestivus

Pantherophis bairdi

—

co)

Pantherophis emoryi

—

Pituophis catenifer

Pituophis deppei*

—

Rhinocheilus lecontei

Salvadora deserticola

Salvadora grahamiae

Sonora episcopa

Tantilla atriceps

Tantilla cucullata

Tantilla gracilis

Tantilla hobartsmithi

Tantilla nigriceps

Tantilla wilcoxi

Trimorphodon vilkinsonii

Dipsadidae (4 species)

Diadophis punctatus

Heterodon kennerlyi

Hypsiglena jani

Leptodeira septentrionalis

Elapidae (1 species)

Micrurus tener

Leptotyphlopidae (3 species)

Rena dissecta

Rena dulcis

Rena segrega

Natricidae (7 species)

Nerodia erythrogaster

Nerodia rhombifer

Storeria hidalgoensis*

Thamnophis cyrtopsis

Thamnophis exsul*

Thamnophis marcianus

Thamnophis proximus

+) 4+]+ + |+ + + +/+ ]+ + ee iat [ete + +] +/+ + +/+) +) +] + +/+ areal |

+] + + + +] +] + + + +] + +] + =

+ + +] + + + +] + +|+ oe

ine]

in|

+] + + +])4+]+ + + + +] + +|+ + w

+ + + +] + + +/+]/+]+ +|+

Crotalus molossus

Amphib. Reptile Conserv. 49 October 2019 | Volume 13 | Number 2 | e189

The herpetofauna of Coahuila, Mexico

Table 4 (continued). Distribution of the amphibians, squamates, and turtles of Coahuila, Mexico, by physiographic region.

Abbreviations are as follows: BDM = Bolson de Mapimi; LS V = Llanuras y Sierras Volcanicas; LDM = Laguna de Mayran; SLC =

Sierras y Llanuras Coahuilenses; SDB = Serranias del Burro; SLP = Sierra de la Paila; PSP = Pliegues Saltillo Parras; STR = Sierras

Transversales; GSP = Gran Sierra Plegada; and LCN = Llanuras de Coahuila y Nuevo Leon. * = species endemic to Mexico; ** =

Species endemic to Coahuila; and *** = non-native species. See text for detailed descriptions of these regions.

Physiographic Regions of Coahuila

Taxa

Crotalus pricei

Crotalus scutulatus

Crotalus viridis

Sistrurus tergeminus

Testudines (13 species)

Emydidae (6 species)

Pseudemys gorzugi

Terrapene coahuila**

Terrapene ornata

Trachemys gaigeae

Trachemys scripta***

Trachemys taylori**

Kinosternidae (3 species)

Kinosternon durangoense*

Kinosternon flavescens

Kinosternon hirtipes

Testudinidae (2 species)

Gopherus berlandieri

Apalone spinifera

borders the US state of Texas, it is not surprising that

all 18 of these species, including the introduced species

Hemidactylus turcicus, also are distributed in the USA.

Of the 143 species comprising the Coahuilan

herpetofauna, 91 (63.6%) are found in only one or two

physiographic regions, which is of great conservation

significance (see below). The mean regional occupancy

is 3.5.

Single-region species: Limited distribution increases

conservation concern. The number of species found ina

single region varies from one (in the Laguna de Mayran,

Llanuras y Sierras Volcanicas, and Pliegues Saltillo-

Parras) to 23 (in the Sierras y Llanurus Coahuilenses). No

single-region species are found in the Serranias del Burro

region. On the following lists, * = endemic to Mexico,

but found in more than one state; and ** = endemic to

Coahuila.

Of the 23 single-region species in the Sierras y

Llanuras Coahuilenses listed here by taxonomic order, 17

are Mexican non-endemics and the other six are endemic

only within the boundaries of Coahuila.

Craugastor augusti

Dryophytes arenicolor

Gerrhonotus lugoi**

Amphib. Reptile Conserv.

Number

of Regions

Occupied

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Gerrhonotus mccoyi**

Gambelia wislizenii

Coleonyx reticulatus

Scincella kikaapoa**

Scincella lateralis

Coluber constrictor

Opheodrys aestivus

Pantherophis bairdi

Salvadora grahamiae

Tantilla atriceps

Tantilla hobartsmithi

Leptodeira septentrionalis

Rena dissecta

Thamnophis cyrtopsis

Crotalus ornatus

Crotalus viridis

Sistrurus tergeminus

Terrapene coahuila**

Trachemys taylori**

Apalone atra**

Of the 15 single-region species in the Gran Sierra

Plegada listed here, 13 are country endemics and the

other two are Mexican non-endemics.

Rheohyla miotympanum*

October 2019 | Volume 13 | Number 2 | e189

Lazcano et al.

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October 2019 | Volume 13 | Number 2 | e189

Amphib. Reptile Conserv.

The herpetofauna of Coahuila, Mexico

Smilisca baudinii

Aquiloeurycea galeanae*

Aquiloeurycea scandens*

Chiropterotriton priscus*

Barisia imbricata*

Gerrhonotus parvus*

Sceloporus minor*

Plestiodon dicei*

Scincella silvicola*

Lampropeltis leonis*

Storeria hidalgoensis*

Thamnophis exsul*

Crotalus morulus*

Crotalus pricei

All of the 12 single-region species in the Llanuras de

Coahuila y Nuevo Leon listed here are non-endemic to

Mexico:

Eleutherodactylus cystignathoides

Acris blanchardi

Crotaphytus reticulatus

Holbrookia lacerata

Sceloporus marmoratus

Masticophis schotti

Salvadora deserticola

Tantilla cucullata

Tantilla gracilis

Tantilla nigriceps

Trimorphodon vilkinsonii

Nerodia rhombifer

Six of the seven single-region species in the Sierras

Transversales listed here are country endemics and the

other is a non-endemic:

Ambystoma velasci*

Sceloporus cautus*

Sceloporus goldmani*

Sceloporus samcolemani*

Sceloporus spinosus*

Xantusia extorris*

Kinosternon hirtipes

Three of the five single-region species in the Bolson

de Mapimi are country endemics, and the other two are

state endemics:

Sceloporus gadsdeni**

Uma exsul**

Uma paraphygas*

Kinosternon durangoense*

Gopherus flavomarginatus*

The single-region species in the Laguna de Mayran is

a state endemic lizard:

Amphib. Reptile Conserv.

Crotaphytus antiquus

The one single-region species in the Llanuras y Sierras

Volcanicas is a non-endemic turtle:

Terrapene ornata

The one single-region species in the Pliegues Saltillo-

Parras is a non-endemic snake:

Tantilla wilcoxi

Examination of the above-listed species indicates that

of the 65 single-region species in Coahuila, 22 are country

endemics and nine are state endemics. The remaining 34

are non-endemic species that are also distributed in the

USA.

Coefficient of Biogeographic Resemblance. A

Coefficient of Biogeographic Resemblance (CBR)

matrix was created for examining herpetofaunal

relationships among the 10 physiographic regions of

Coahuila (Table 6) and these data were used to produce a

UPGMA dendrogram (Fig. 12). As mentioned above, the

numbers of species within the 10 physiographic regions

of Coahuila range from a high of 91 species within the

Sierras y Llanuras Coahuilenses (SLC) to a low of 38

within Laguna de Mayran (LDM). The mean species

richness number for all 10 regions is 50.3. The numbers

of species shared between regions range from 20 to 45.

The lowest value of 20 is found between only one pair of

regions, the Llanuras de Coahuila y Nuevo Leon (LCN)

and Gran Sierra Plegada (GSP). The highest number is

also shared between only one pair of regions, the SLC

and Serranias del Burro (SDB). The mean number of

shared species among the 45 regional pairings 1s 32.9.

The lowest number of 20 shared species between LCN

and GSP makes biogeographic sense because these two

regions are situated at opposite ends of the state, are not

connected geographically, and are environmentally quite

different: LCN contains subhumid lowland plains and

hills versus GSP with semihumid to subhumid highland

mountainous areas carved by deep valleys. Also, GSP has

a much smaller area in Coahuila than does LCN. On the

other hand, the two regions with the highest number of

AS shared species are SLC and SDB. These two regions

share part of their borders and both contain similar

ecological regimes. Unlike the situation in Tamaulipas

(Teran-Juarez et al. 2016), the higher numbers of species

in the regional pairings in Coahuila, with the exception

of SLC (91 species), do not necessarily equate to higher

numbers of shared species, which is more similar to the

patterns shown in adjacent Nuevo Leon (Navarez-de

los Reyes et al. 2016). This discrepancy is most likely

due to the larger number of included physiographic

regions, and the lower number of shared species from

more distant regions. Reflecting this trend, the following

October 2019 | Volume 13 | Number 2 | e189

Lazcano et al.

Table 6. Pair-wise comparison matrix of Coefficient of Biogeographic Resemblance (CBR) data of herpetofaunal relationships for

the 10 physiographic regions in Coahuila, Mexico. Underlined values = number of species in each region; upper triangular matrix

values = species in common between two regions; and lower triangular matrix values = CBR values. The formula for this algorithm

is CBR = 2C/N, + N, (Duellman 1990), where C is the number of species in common to both regions, N, is the number of species

in the first region, and N, is the number of species in the second region. See Table 4 for explanation of abbreviations and Fig. 12 for

the UPGMA dendrogram produced from the CBR data.

BDM LSV LDM SLC

BDM 45 38 af 38

LSV 0.85 44 37 42

LDM 0.89 0.90 33 ey

SLC 0.56 0.62 0.57 91

SDB 0.76 0.81 0.82 0.66

SLP 0.85 0.88 0.92 0.61

PSP 0.79 0.80 0.83 0.63

STR 0.72 0.70 0.75 0.49

GSP 0.60 0.61 0.65 0.44

LCN 0.45 0.45 0.48 0.56

SDB SLP PSP STR GSP LCN

34 36 37 33 29 22

36 37 36 32 29 22

34 36 36 32 I) 22

45 40 4 34 31 40

45 35 37 30 27 28

0.74 40 38 34 29 24

0.79 0.85 49 35 32 24

0.65 0.78 0773 47 31 21

0.56 0.64 0.64 0.63 Si 20

0.57 0.52 0.47 0.42 0.38 53

pairwise comparisons of regions are aligned in order of

highest to lowest species richness (underlined values)

and their corresponding numbers of shared species (in

parentheses) with all other regions; see text and map for

discussions on characteristics and sizes of the regions:

SLC 91: SDB (45), SLP (40), PSP (44), STR (34),

GSP (31), LCN (40), LDM (32), LSV (42), BDM (38).

LCN 53: GSP (20), STR (21), PSP (24), SLP (24),

SDB (28), SLC (40), LDM (22), LSV (23), BDM (22).

GSP 51: LCN (20), STR (31), PSP (32), SLP (29),

SDB (27), SLC (31), LDM (29), LSV (29), BDM (29).

PSP 49: STR (35), GSP (32), LCN (24), SLP (38),

SDB (37), SLC (44), LDM (36), LSV (36), BDM (37).

STR 47: GSP (31), LCN (21), PSP (35), SLP (34),

SDB (30), SLC (34), LDM (32), LSV (32), BDM (33).

SDB 45: SLP (35), PSP (27), STR (30), GSP (27), LCN

(28); SLC (45), LDM (34), LSV (36), BDM (34).

BDM 45: LSV (38), LDM (37), SLC (38), SDB (34),

SLP (36), PSP (37), STR (33), GSP (29), LCN (22).

LSV 44: BDM (38), LDM (37), SLC (42), SDB (36),

SLP (37), PSP (36), STR (32), GSP (29), LCN (22).

SLP 40: BDM (36), LSV (37), LDM (36), SLC (40),

SDB (35), PSP (38), STR (34), GSP (29), LCN (24).

LDM 38: BDM (37), SLP (36), PSP (36), STR (32),

GSP (29), LCN (22), SLC (37), SDB (34), LSV (34).

SLC, with its 91 species, is the largest physiographic

region in Coahuila that shares borders to variable extents

with five of the nine other regions in the state (LCN,

SDB, PSP, LSV, SLP), including the 2™ and 4" most

speciose regions (LCN, PSP). The 91 species in SLC

reveals a large discrepancy between it and all nine other

regions in the state. Ninety-one species is 38 more than

found in LCN, the second most species-rich region with

53 species, whereas the total difference for all nine of the

Amphib. Reptile Conserv.

other regions is only 15 species between the 53 species in

LCN and the 38 species in LDM. LCN is a lowland region

next to the Rio Grande with few montane landscapes, but

it contains several generalist herpetofaunal species that

also exist in adjacent montane regions of Coahuila at

lower elevations. PSP is mostly separated from LCN by

two other regions to its north (LDM, SLP); however, it

shares a geographic connection through a northwestern

extension of PSP in Nuevo Leon (Nevarez-de los Reyes

et al. 2016).

The following data show ranges and mean numbers

of shared species for each of the 10 regions listed above

that are arranged according to increasing mean numbers

(bold in parentheses) with underlined values referring to

species richness in each region:

Llanuras de Coahuila y Nuevo Leon (LCN) (53): 20-—

40 (24.7)

Gran Sierra Plegada - GSP (51): 20-32 (28.5)

Sierras Transversales - STR (47): 21-35 (31.3)

Laguna de Mayran - LGM (38): 22-37 (33.3)

Bolson de Mapimi - BDM (45): 22—38 (33.8)

Serranias del Burro - SDB (45): 27-45 (34.0)

Llanuras y Sierras Volcanicas - LSV (44): 22-42 (34.3)

Sierra de la Paila - SLP (40): 2440 (34.3)

Pliegues de Saltillo Parras - PSP (49): 24-38 (35.4)

Sierras y Llanuras Coahuilenses - SLC (91): 31-45 (39.0)

With the exception of SLC and PSP (1* and 4" highest

in species richness, and 1%‘ and 2" highest mean numbers

of shared species, respectively), the mean number of

pairwise species comparisons between all other regions

indicate that higher species richness in a region does not

necessarily translate into a higher mean number of shared

species when all regions are totaled. Apparent extreme

examples of this are: LCN, GSP, and STR, respectively,

having the 1%, 2" and 3™ highest numbers of species

and lowest mean numbers of shared species. It makes

sense that LCN would share fewer species with other

regions in Coahuila because of its ecological uniqueness

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The herpetofauna of Coahuila, Mexico

Fig. 9. Sierras y Llanuras Coahuilenses. Vegetation in the Val-

ley of Cuatrociénegas, in the municipality of Cuatrociénegas de

Carranza. Photo by Eli Garcia-Padilla.

Fig. 11. Sierras Transversales. Paso de Carneros, Matorral ro-

setophilous vegetation, with Chocha (Yucca carnerosana) and

Lechuguilla (Agave lechuguilla), at Paso de Carneros, in the

municipality of Saltillo. Photo by Manuel Nevarez de los Reyes.

associated with lower elevations and general differences

in vegetation formations and topography, as well as

herpetofaunal affinities to the United States northward

across the Rio Grande. GSP has the smallest area of all

regions in the state, but is much more extensive when

considering it also exists in Nuevo Leon and Tamaulipas

(Nevarez-de los Reyes et al. 2016; Teran-Juarez et al.

2016). STR is a slender montane region with high species

richness positioned primarily within Coahuila across its

entire southern border.

UPGMA Dendrogram. Based on the data in Table 6, a

UPGMA dendrogram (Fig. 12) was created to illustrate

the herpetofaunal resemblance patterns in a hierarchical

fashion among the 10 physiographic regions of Coahuila

(Fig. 1). The patterns are different when compared to

those shown in the two other northern Mexico states

bordering Texas that were covered in previous MCS

publications: Nuevo Leon (Nevarez-de los Reyes et al.

2016) and Tamaulipas (Teran-Juarez et al. 2016). The

Coahuila dendrogram shows the similarity relationships

in descending order from the most similar regions,

SLP clustering with LDM at a value of 0.92, down to

the lowest value, where LCN clusters with all the other

regions at a value of 0.48. In other words, there are no

Amphib. Reptile Conserv.

Fig. 10. Sierra de la Paila. Vegetation in the Sierra de la Paila,

in the municipality of Ramos Arizpe. Photo by Bernardo Ma-

rino (http://gransierraplegada.org).

distinct subgroupings within the dendrogram, which

indicates on a biogeographic scale that there are no

distinct subgroups composed of distributional units that

share more closely related herpetofaunas. On the other

hand, neighboring Nuevo Leon with seven regions

has two distinct biogeographic subgroups, a southern

unit containing two regions and a more northern unit

containing five regions that clusters with the southern

unit at the 0.37 similarity level. Tamaulipas, also with

seven recognized regions, has the most complex pattern

of herpetological similarity of these three states. Two

biogeographic subgroups are found in what can be

considered the northern and eastern sections of the

state. One of those subgroups contains three regions that

make up the majority of the state’s area, while the other

subgroup is comprised of two small disjunct highland

regions that cluster together at the 0.55 level; and both

are nested within one of the other subgroup’s regions.

Those two subgroups cluster with each other at the 0.46

similarity level. One of the two remaining regions, which

make up the extreme southwestern sector of Tamaulipas,

clusters independently with the other two biogeographic

subgroups at the 0.44 level of herpetofaunal similarity.

The last region, which is the southwestern-most

section of the state, 1s the most distinctive of the seven,

thereby clustering with the others at the 0.23 level of

herpetological similarity.

In summary, the UPMGA dendrogram for Coahuila

shows that the lowland non-montane region (LCN)

bordering the Rio Grande and Texas is the most

distinctive region in Coahuila as far as herpetological

similarity goes, based on numbers of shared species.

It also shows a pattern of similarity among regions in

close proximity to each other that also share ecological

parameters either within the state or through areas of the

same physiographic region outside Coahuila.

Distribution Status Categorizations

The discussion of the distribution status of Coahuilan

herpetofauna members uses the system developed by

Alvarado-Diaz et al. (2013), and employed in all the

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Lazcano et al.

SLC GSP

LCN STR SDB

PSP

BDM LSV SLP LDM

Fig. 12. AUPGMA generated dendrogram illustrating the similarity relationships of species richness among the herpetofauna in the

10 physiographic regions of Coahuila (based on the data in Table 6). We calculated the similarity values using Duellman’s (1990)

Coefficient of Biogeographic Resemblance (CBR).

other entries in the Mexican Conservation Series. The

categories in the system are non-endemic, country

endemic, state endemic, and non-native (Tables 7 and 8).

Given the 512-km-long border shared between

Coahuila and Texas (http://wikipedia.org; accessed

11 August 2018), it is not surprising that the largest

component of the herpetofauna falls into the non-

endemic category. Of the 143 species comprising the

Coahuilan herpetofauna, 100 species (69.9% of the total)

belong to this category. Almost half (49) of the non-

endemic species are snakes, and this number is only five

fewer than the total number of snake species in the entire

herpetofauna (Table 8). The 76 non-endemic squamate

species are 71.7% of the total of 106 species for the state.

In addition, a large portion of the state’s anurans (17 of

20; 85.0%) are also non-endemic species. On the other

hand, slightly more than half the turtle species (seven of

13; 53.8%) are non-endemic to Coahuila (Table 8).

The next largest component is the 31 (21.7%) country

endemic species, most of which are squamates, including

18 lizards and five snakes (74.2%). The remainder

are amphibians (six species; 19.4%) and turtles (two

species; 6.5%). Almost half of the country endemics are

phrynosomatid lizards (14 of 31; 45.2%).

Only nine of the species (6.3%) in Coahuila are state

endemics. Six of these are lizards (Gerrhonotus lugoi,

G. mccoyi, Crotaphytus antiquus, Sceloporus gadsdeni,

Uma exsul, and Scincella kikaapoa) and three are turtles

(Terrapene coahuila, Trachemys taylori, and Apalone

atra).

The number of non-native species in Coahuila is

only three, the ranid frog Lithobates catesbeianus, the

Amphib. Reptile Conserv.

gekkonid lizard Hemidactylus turcicus, and the emydid

turtle Trachemys scripta. These three species also were

reported as introduced into Nuevo Leon (Nevarez-de los

Reyes et al. 2016).

The number of endemic species in Coahuila (country

and state endemics combined) is 40, which is 4.9% of

the total number of endemic species for Mexico (811;

Johnson unpub.). The number of non-endemic species is

100, which is 19.7% of the total of such species in the

entirety of Mexico (508; Johnson unpub. ).

Principal Environmental Threats

In this section we examine the 12 problems we think are

the most significant in affecting the sustainability of the

populations of Coahuila’s amphibians and reptiles.

Urban development. As of 2015, the population of

Coahuila was 2,954,915, making it the 16" most densely

populated state in Mexico. The Municipality of Saltillo is

the most populated of the 38 municipalities in the state,

with a population of 807,537 people, followed by the

municipalities of Torredn with 679,288, Monclova with

231,107, and Piedras Negras with 163,595 (see INEGI,

http://www. beta.inegi.org.mx/temas/estructura/). Data

from this same website indicate that 90% of the state

population is located in urban areas, with the remainder

in rural areas. The current annual percentage growth

rate 1s 1.5%, which portends a doubling rate of about 47

years. Most of this growth is expected to occur within the

most heavily-populated municipalities.

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The herpetofauna of Coahuila, Mexico

Table 7. Distributional and conservation status measures for members of the herpetofauna of Coahuila, Mexico. Distributional

Status: SE = endemic to state of Coahuila; CE = endemic to country of Mexico; NE = not endemic to state or country; and NN

= non-native. Environmental Vulnerability Score (taken from Wilson et al. 2013a,b): low (L) vulnerability species (EVS of 3-9);

medium (M) vulnerability species (EVS of 10—13); and high (H) vulnerability species (EVS of 14—20). IUCN Categorization: CR =

Critically Endangered; EN = Endangered; VU = Vulnerable; NT = Near Threatened; LC = Least Concern; DD = Data Deficient; NE

= Not Evaluated. SEMARNAT Status: A = Threatened; P = Endangered; Pr = Special Protection; and NS = No Status. * = species

endemic to Mexico; ** = species endemic to Coahuila; *** = non-native species. See text for explanations of the EVS, IUCN, and

SEMARNAT rating systems.

Datributionale ||| oo ronmental IUCN SEMARNAT

Vulnerability aecid

Status Category (Score) Categorization Status

| Lithobates catesbeianus***_ | NNT OS

| Hemidactylus turcicus*** | NN

Amphib. Reptile Conserv. 56 October 2019 | Volume 13 | Number 2 | e189

Lazcano et al.

Table 7 (continued). Distributional and conservation status measures for members of the herpetofauna of Coahuila, Mexico.

Distributional Status: SE = endemic to state of Coahuila; CE = endemic to country of Mexico; NE = not endemic to state or country;

and NN = non-native. Environmental Vulnerability Score (taken from Wilson et al. 2013a,b): low (L) vulnerability species (EVS of

3-9); medium (M) vulnerability species (EVS of 10-13); and high (H) vulnerability species (EVS of 14—20). IUCN Categorization:

CR = Critically Endangered; EN = Endangered; VU = Vulnerable; NT = Near Threatened; LC = Least Concern; DD = Data

Deficient; NE = Not Evaluated. SEMARNAT Status: A= Threatened; P = Endangered; Pr = Special Protection; and NS = No Status.

* = species endemic to Mexico; ** = species endemic to Coahuila; *** = non-native species. See text for explanations of the EVS,

IUCN, and SEMARNAT rating systems.

Distributional || vironmental IUCN SEMARNAT

Vulnerability ae

Status Category (Score) Categorization Status

| Umaexsul** TSE HS) | ENT |

| Umaparaphygas* | CE) | NTP

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The herpetofauna of Coahuila, Mexico

Table 7 (continued). Distributional and conservation status measures for members of the herpetofauna of Coahuila, Mexico.

Distributional Status: SE = endemic to state of Coahuila; CE = endemic to country of Mexico; NE = not endemic to state or country;

and NN = non-native. Environmental Vulnerability Score (taken from Wilson et al. 2013a,b): low (L) vulnerability species (EVS of

3-9); medium (M) vulnerability species (EVS of 10—13); and high (H) vulnerability species (EVS of 14—20). IUCN Categorization:

CR = Critically Endangered; EN = Endangered; VU = Vulnerable; NT = Near Threatened; LC = Least Concern; DD = Data

Deficient; NE = Not Evaluated. SEMARNAT Status: A = Threatened; P = Endangered; Pr = Special Protection; and NS = No Status.

* = species endemic to Mexico; ** = species endemic to Coahuila; *** = non-native species. See text for explanations of the EVS,

IUCN, and SEMARNAT rating systems.

Taxa Status Sy HEE AUILY, Categorization Status

Category (Score) 8

| Trachemys scripta*** | NNT

| Kinosternon durangoense* | CCE OT CO) S|

Amphib. Reptile Conserv. 58 October 2019 | Volume 13 | Number 2 | e189

N Ls

= |

SS)

Lazcano et al.

Fig. 13. Urban Development. Urban growth near Saltillo,

reaching the limit of the Natural Protected Area “Sierra de

Zapalinamé,” in the municipality of Saltillo. Photo by Manuel

Nevarez de los Reyes.

eee oe ee

Fig. 15. Deforestation for Agricultural Purposes. Monoculture

of grapes near Parras de la Fuente, in the municipality of Parras.

Photo by Manuel Nevarez de los Reyes.

Urban pollution. One outcome of urban growth that

is of environmental significance is the accumulation of

garbage resulting from the inefficient use of resources

by populations in urban areas. An extensive recent study

acknowledged that “garbage is the major environmental

problem facing Mexico, involving the generation of more

than 100 million tons of wastes per year that are not

handled in an adequate manner” (http://www.estosdias.

com.mx/blog/archivos/226; accessed 18 September

2018). This site points out that the Federal District and

its metropolitan area has the largest garbage dump in the

world, located in Ciudad Nezahualcoyotl in the State of

México. The useful life of this site has been extended

despite the lack of additional space being available,

which points out the difficulty of finding other sites to

deposit the thousands of tons of garbage produced.

With reference to Coahuila, in an article published in

Torreon on 14 August 2018 by Noticias—El Sol de la

Laguna, the Secretary of the Environment, Eglantina

Canales Gutiérrez, reported that 3,000 tons of garbage

are produced in the state of Coahuila on a daily basis,

or almost one kilogram of trash produced per person

Amphib. Reptile Conserv.

Fig. 14. Industrial Pollution. Factories polluting the air in the

vicinity of Monclova, in the muncipality of Monclova. Photo

by Michael Price.

Fig. 16. Deforestation for Agricultural Purposes. Monocul-

ture of cotton at San Pedro de las Colonias, in the municipality

of the same name, Provincia de Laguna de Mayran. Photo by

Manuel Nevarez de los Reyes.

per day. Secretary Canales stated that the burial of such

trash in landfills, which are available in 85% of Coahuila,

represents the best solution to date, but that in the future

efforts to recycle products should be implemented. She

decried that often garbage does not end up tn landfills,

but rather is left out in the open. This environmental

problem can be expected to grow commensurate with the

rate of human population growth in Coahuila.

Several instances of the direct impact of accumulated

garbage on members of the Mexican herpetofauna have

been documented. Lazcano et al. (2006) reported the

death of several Texas Horned Lizards (Phrynosoma

cornutum), that were trapped inside a discarded tire in

an illegal dump site in the neighboring state of Nuevo

Leon. The lizards, presumably seeking shelter, died

after they were unable to escape due to the intense daily

temperatures at this locality that can rise to 45 °C in the

shade. On another occasion in Nuevo Leon, Chavez-

Cisneros et al. (2010) reported finding a Greater Earless

Lizard (Cophosaurus texanus) that apparently died after

ingesting a deflated balloon left in a pile of litter. The full

environmental impact of discarded trash on the native

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The herpetofauna of Coahuila, Mexico

Table 8. Summary of the distributional status of herpetofaunal families in Coahuila, Mexico.

Distributional Status

Number of

Families ;

Species

Hylidae

Microhylidae

Ranidae

Scaphiopodidae

Subtotals

Plethodontidae

Sphenomorphidae

Teiidae i

Xantusiidae

Dipsadidae

Elapidae

Leptotyphlopidae

Natricidae

Viperidae

Subtotals

Emydidae

Kinosternidae

Sum Totals

herpetofauna of Coahuila, and elsewhere in Mexico, is

still unknown, so it 1s imperative that actions be taken to

diminish this overtly intentional behavior by an uncaring

populace.

Industrial pollution. The municipalities of Acufia,

Monclova, Piedras Negras, Ramos Arizpe, Saltillo, and

Torreon have a Vehicle Verification Program (PVV) to

address vehicular air pollution, which is supervised

under municipal authority (Anon 2017).

Coahuila produces 16% of the electric power and

23% of the steel in Mexico. The industry dedicated

to the extraction and commercialization of coal, in

the coal region around Nava and Piedras Negras, has

Amphib. Reptile Conserv.

Non-endemic

Country

Endemic

(CE)

State Non-native

Endemic (SE)

been named as the one that most intensively pollutes

the air (Journalistic note of May 4, 2016, http://www.

vanguardia.com.mx/articulo/empresas-carboneras-las-

que-mas-contaminan-el-aire-en-la-carbonifera-y-norte-

de-coahuila).

Deforestation for agricultural and ranching purposes.

One of the most notorious cases of massive deforestation

carried out in the state of Coahuila happened in 2001.

It occurred in the Valle del Hundido, adjacent to the

Cuatro Ciénegas Valley, where hundreds of hectares

were cleared for the establishment of alfalfa crops to

feed dairy and beef cattle (http://www.vanguardia.com.

mx/columnas-elhundido-1728357.html). Another case

October 2019 | Volume 13 | Number 2 | e189

Lazcano et al.

== : 7 a ee | = ~ = ‘\ : —e a a

No. 9. Phrynosoma orbiculare (Linnaeus, 1758). The Mountain Horned Lizard is a Mexican endemic species that occurs “from

eastern Sonora and western Chihuahua southward through the mountains of Durango, Zacatecas, Aguascalientes, Jalisco, and

Michoacan, and from the mountains of southern Nuevo Leon southward through San Luis Potosi, Querétaro, Hidalgo, Veracruz,

and westward through Puebla, Tlaxcala, Mexico, the Distrito Federal, and Morelos” (Lemos-Espinal and Dixon 2013: 122). Bryson

et al. (2011) noted, however, that this species is probably comprised of several distinct lineages, of which some appear to have

small distributions and long independent evolutionary histories, and that some of these lineages merit additional consideration for

protection. This individual was found near Monterreal, in the municipality of Arteaga. Wilson et al. (2013a) calculated its EVS as

12, placing it in the upper half of the medium vulnerability category. Its conservation status has been reported as Least Concern by

IUCN, and as threatened (A) by SEMARNAT. Photo by Eli Garcia-Padilla.

~ Rat r o /. »

.- = fe tom a” ‘. = s Ae > * a

No. 10. Sceloporus cautus Smith, 1938. The Shy Spiny Lizard is an endemic Mexican lizard distributed from “the western slopes

of the Sierra Madre Oriental in Tamaulipas, southeastern Coahuila, and central Nuevo Leon southward through much of San Luis

Potosi and the northern half of Zacatecas” (Lemos-Espinal and Dixon 2013: 124). This individual was found at Cafion el Chorro, in

the municipality of Arteaga. Wilson et al. (2013a) estimated its EVS as 15, placing it in the lower portion of the high vulnerability

category. Its conservation status has been gauged as Least Concern by IUCN, but it is not listed by SEMARNAT. Photo by Michael

S. Price.

Amphib. Reptile Conserv. 61 October 2019 | Volume 13 | Number 2 | e189

The herpetofauna of Coahuila, Mexico

No. 11. Sceloporus olivaceus Smith, 1934. The Texas Spiny Lizard is found from “northern central Texas southward through

the Gulf of Mexico coastal plain to southern Tamaulipas, westward nearly to the Big Bend area of Texas and eastern Coahuila”

(Lemos-Espinal et al. 2015: 239). This individual was encountered at Cafion el Chorro, in the municipality of Arteaga. Wilson et al.

(2013a) calculated its EVS as 13, placing it at the upper limit of the medium vulnerability category. Its conservation status has been

considered as Least Concern by IUCN; this species is not listed by SEMARNAT. Photo by Michael S. Price.

b a ie ys was e wa \e her a : A) ae Bt be > ‘

No. 12. Sceloporus ornatus Baird, 1859. The Ornate Spiny Lizard is a Mexican endemic species distributed in “southern and central

Coahuila” (Lemos-Espinal and Smith 2007: 303). This individual was found at ca. 8 km SW from Ejido Mayran, in the municipality

of San Pedro. Wilson et al. (2013a) determined its EVS as 16, placing it in the middle portion of the high vulnerability category.

Its conservation status has been calculated as Near Threatened by IUCN and as threatened (A) by SEMARNAT. Photo by Marco

Antonio Bazdn-Tellez.

Amphib. Reptile Conserv. 62 October 2019 | Volume 13 | Number 2 | e189

Lazcano et al.

involved members of the Mennonite community in

Coahuila who were implicated in the clearing of 2,300

hectares of forest vegetation without authorization in the

municipality of Sierra Mojada; however, they managed

to escape legal sanction (https://www.eluniversal.com.

mx/estados/menonitas-de-coahuila-ganan-amparos-la-

profepa-por-2-mil-hectareas-de-predios).

Effect of roads. The state of Coahuila has an extensive

network of roads and highways, with a total of seven

federal highways. The total length of the road network

in the state is 8,336 km (Servicio Geolégico Mexicano

2017). During the period from 1980 to 2015, the

number of motor vehicles traveling within Coahuila

went from 149,242 to 741,515, an increase of 592,273

(or almost 400%!) in 35 years. The effect of roads on

the herpetofauna of the state remains to be studied, but

we assume that major highways might disrupt general

dispersion patterns and seasonal migration of some local

populations, and vehicles on municipal roads probably

simply run over many animals in large numbers.

Mining and energy projects. The history of Coahuila

is closely related to mining. It began as a leading

commercial activity in the colonial era, with the founding

of the city of Monclova and Minas de la Trinidad in 1577,

and later continued with the discovery and exploitation of

coal starting in 1828; copper in the Panuco mine in 1870;

zinc oxide, silver, and lead in Sierra Moyjada in 1879; and

silver, lead, and zinc in Reforma-Santa Teresa in 1890.

Most recently, the discovery and exploitation of fluorite,

celestite, sodium-magnesium salts, gypsum, barite, and

dolomite has been undertaken. Coahuila has an extensive

mining-metallurgical infrastructure, highlighted by the

metal foundry in Torreon, the iron foundry in Monclova,

coal plants in Nava, and several related processing plants

in various locations. The state of Coahuila contributed

3.1% of the value of national mining production in 2015,

occupying the first place in the production of iron, coal,

celestite, magnesium sulfate, sodium sulfate, bismuth,

and cadmium, second place in fluorite and silica, third

in barite and dolomite, fifth in stone aggregates, and in

smaller proportions plaster, sulfur, clays, gravel, sand,

limestone, gold, and silver. In addition, a large number

of unexplored geological areas have been reported in

Coahuila (Servicio Geolégico Mexicano 2017), for

example, the Hercules mine in the northwestern portion

of the state (http://www.thediggings.com/mines/2 1966;

accessed 20 July 2019), and those places are expected to

be sites for future development.

Natural gas production in the Burgos Basin. The

Burgos Basin is a shale deposit located in the northeastern

portion of Coahuila, directly south of the Rio Grande. It

is considered a great prospect for natural gas production

and covers a total area of 62,677 km’. The infrastructure

required to develop the production of natural gas is

Amphib. Reptile Conserv.

scheduled to be built in a timely manner during successive

10-year periods, and involves extensive extraction,

processing, and distribution facilities. It is expected that

the previous and on-going infrastructure projects will

have negative effects on the integrity of the habitat that

is used by the members of the regional herpetofauna

(http://energiaadebate.com/gas-de-lutitas-en-la-cuenca-

de-burgos/).

Wind turbines. In 2016, a wind farm consisting of 95

wind turbines was being built near Ejido Hipolito in the

Municipality of Ramos Arizpe, and another with 100

wind towers near Acufia, as well as a Solar Energy Park

in the Municipality of Viesca, which will be the largest in

Latin America and is planned to include more than two

million solar panels (http://coahuila.gob.mx/noticias/

index/avanza-parque-eolico-de-hipolito-24-07-16). The

Ramos Arizpe project covers an area of 4,754 hectares.

This requires, among other actions, the construction of

more than 50 km of access roads, an aerial transmission

line almost five km long, and an electrical substation

(http://proyectoeolicadecoahuila.com).

Elimination of herpetofauna due to cultural beliefs

and practices. Traditional beliefs and practices that

affect the herpetofauna of Coahuila are the same as

those previously reported for Nuevo Leon (Nevarez-de

los Reyes et al. 2016), likely due to their geographic

proximity and similar cultural backgrounds. Examples

include snakes being slaughtered due to fear and

superstition, misconceptions that many non-venomous

herpetofaunal species are venomous, and rattlesnakes

being consumed either for food or the belief that their

meat will prevent or cure cancer. It is obvious that

education is the key for reducing the needless killing of

these ecologically important denizens of Coahuila.

Use of pesticides. Agricultural activities in the state of

Coahuila are most intense in the arid region known as

Comarca Lagunera, where they are only possible due

to heavy irrigation using water from the Rio Nazas.

Although there is little information regarding this

problem, pesticides are commonly applied on Cantaloupe

(Cucumis melo) fields, one of the most important fruits

cultivated in the region (Vargas-Gonzalez et al. 2016).

That study found that in the agricultural cycle of 2010,

50 different active ingredients were used in the region;

26% of which were not authorized for use on cantaloupes

by COFEPRIS, and 46% of which were considered as

highly toxic to human health and the environment.

For example, six of them (carbofuran, endosulfan,

clorotalonil, mancozeb, imidacloprid, and metamidofos)

are regarded as among the most toxic to humans and the

environment.

Collecting and commercial trade. Currently, the

herpetofauna of Coahuila faces problems with unlawful

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The herpetofauna of Coahuila, Mexico

collecting and commercial trade that are similar to those

reported in Tamaulipas and Nuevo Leon (Teran-Juarez et

al. 2016; Nevarez-de los Reyes et al. 2016). Unfortunately,

these illegal activities have increased during the last few

years through social media. For example, based on our

personal observations, when photographs of rare species

are posted, it is common for collectors to be immediately

contacted by animal traffickers or pet owners ready to

purchase their specimens, or asked for specific localities.

Unfortunately, monitoring these activities remains

difficult, as does determining their true impact on the

local herpetofauna.

Fires involving natural habitats. Fires create a serious

threat to natural habitats in Coahuila, especially those

involving forested ecosystems. In 2017, 153 fires were

documented in the state beginning on 2 January and

ending on 23 December; thus, on average, a reported fire

occurred somewhere in the state every 2.4 days. Of the

38 municipalities in Coahuila, fires were recorded in 22

(57.9%) of them. The municipalities most often involved

were Arteaga (37 fires), Cuatro Ciénegas (12), Muzquiz

(11), Ramos Arizpe (23), and Saltillo (24). Three of these

municipalities (Arteaga, Ramos Arizpe, and Saltillo) are

located in the extreme southeastern portion of the state

where the state capital is located and where a majority of

the fires have occurred (84 of 153 fires, or 54.9%). The

number of ha involved in each of these 153 fires ranged

from 0.02 to 3,132. The largest of these fires took place

in the Municipality of San Buenaventura, at a locality

called, interestingly enough, El Quemado (“burned

by fire’). The total area burned in 2017 amounted to

10,289.6 ha, which is approximately 0.07% the total area

of Coahuila (15,159,500 ha).

At the time of this writing, information is also available

for the first 4.5 months of 2018 (10 January to 13 May)

(CONAFOR, Comisién Nacional Forestal Aspectos de

Incendios Forestales, —https://www.gob.mx/conafor).

During this period of time, 46 fires were reported, or one

fire in Coahuila every 2.9 days. Fires were registered

in 14 of the 38 municipalities in the state (36.8%).

These fires occurred most often in the Municipalities

of Muzquiz (15 fires), Cuatro Ciénegas (four), Arteaga

(three), Nadadores (three), Ocampo (three), and Saltillo

(three). Four of these six municipalities (Muzquiz,

Cuatro Ciénegas, Arteaga, and Saltillo) represent areas

with high fire occurrences during 2017 (see above). The

number of hectares burned in each fire during this 2018

period varied from 0.5 to 1,500. The largest fire took

place in Eutimias, in the Municipality of Ocampo, due to

lightning strikes. In general, the principal causes of these

fires, when known, were human negligence and lightning.

During this period, the total area burned was 4,198.3 ha

(about 0.03% of the state, or 31.3 ha/ day). This figure

compares to 28.2 ha/day in 2017. It appears likely that

the total number of hectares burned in Coahuila in 2018

should compare to those burned in 2017 (see above). The

Amphib. Reptile Conserv.

threat posed by fires will almost assuredly increase into

the future, given the human population growth in the

state of 7.5% from 2010 to 2015, which is higher than

the rate in Mexico as a whole (6.8%; http://wikipedia.

org; accessed 5 July 2018).

Lazcano et al. (2006) reported finding a Wiegmann’s

Alligator Lizard (Gerrhonotus liocephalus) killed by a

forest fire in the neighboring state of Nuevo Leon and

advised that only additional investigations would be

able to determine the demographic consequences of this

sort of mortality. Banda-Leal et al. (2018) documented

for the first time the distribution of Gerrhonotus parvus

in Coahuila, and mentioned finding some specimens of

this species in the Sierra Zapalinamé that had died from

forest fires.

Conservation Status

The discussion of the conservation status of members

of the Coahuilan herpetofauna follows the same three

systems of conservation assessment as used in the other

entries 1n the Mexican Conservation Series. These

systems are those found in SEMARNAT (2010), the

IUCN Red List (http://tucnredlist.org), and the EVS

(Wilson et al. 2013a,b). The assessments from these three

systems were updated as needed.

The SEMARNAT System. SEMARNAT (Secretaria

del Medio Ambiente y Recursos Naturales) is the

environmental ministry of Mexico, and it is “charged with

the mission of protecting, restoring, and conserving the

ecosystems, natural resources, assets and environmental

services of Mexico with the goal of fostering sustainable

development’ (http://www.semarnat.gob.mx/conocenos/

quienessomos; accessed 7 January 2019). In 2010, this

agency published the Norma Oficial Mexicana (Official

Mexican Standard)-059, which deals with the protection

of the native members of the Mexican flora and fauna and

establishes categories of risk. This system is commonly

used by Mexican herpetologists to discuss various

segments of the country’s herpetofauna. Its utility for work

on the herpetofauna of Coahuila has also been assessed.

The SEMARNAT system comprises three categories,

including Endangered (P), Threatened (A), and Special

Protection (Pr). For a Mexican species not evaluated

using one of these three categories, it is designated here

as having no status (NS). The SEMARNAT evaluations

are shown in Table 7 and summarized in Table 9.

Unfortunately, the SEMARNAT assessments are

not very useful here, as most species in Coahuila remain

unevaluated (90 of 140 native species; 64.3%). Thus,

evaluations of conservation status are available for only

50 species (35.7%). Of these 50 species, 23 (46.0% of

the total) are judged as species of Special Protection (Pr):

Anaxyrus debilis, Gastrophryne olivacea, Lithobates

berlandieri, Ambystoma velasci*, Aquiloeurycea

scandens*, Chiropterotriton priscus*, Gerrhonotus

October 2019 | Volume 13 | Number 2 | e189

Lazcano et al.

—— ‘a og = a pen . +e — ae < : :

ee ee “ id ee oe -.

vicinity of Estacién Marte, in the municipality of General

Cepeda. Photo by Manuel Nevarez de los Reyes.

Fig. 19. Impact of Roads. Masticophis flagellum dead on the

road near El Mimbre, in the municipality of Parras. Photo by

Manuel Nevarez de los Reyes.

Fig. 21. Mining Projects. Mining of materials for construction

near Paso de Carneros, in the municipality of Saltillo. Photo by

Manuel Nevarez de los Reyes.

parvus, Gambelia wislizenii, Coleonyx brevis, Coleonyx

reticulatus, Sceloporus grammicus, S. maculosus*,

Scincella lateralis, Crotalus atrox, C. lepidus,C. molossus,

C. pricei, C. scutulatus, C. viridis, Sistrurus tergeminus,

Terrapene ornata, Kinosternon hirtipes, and Apalone

spinifera. These 23 species include four that are endemic

to Mexico (indicated by asterisks); and all of the 19 non-

endemic species are shared with the United States. The

Amphib. Reptile Conserv.

= =

Fig. 18. Deforestation for Ranching Purposes. Cattle in the mu-

nicipality of Francisco I. Madero. Photo by Manuel Nevarez de

los Reyes.

Fig. 20. Mining Projects. Mineral charcoal mining exploitation,

in the municipality of Nava. Photo by Manuel Nevarez de los

Reyes.

Fig. 22. Energy Projects. Wind generation of electricity near

Hipolito, in the municipality of Ramos Arizpe. Photo by Manu-

el Nevarez de los Reyes.

remaining 27 species are assessed as either Endangered

(P) or Threatened (A). The Endangered species

amount to four: Uma exsul, U. paraphygas, Gopherus

flavomarginatus, and Apalone atra (Table 7); three are

endemic to Coahuila and one is endemic to Mexico.

Twenty-three species are considered as Threatened:

Aquiloeurycea galeanae, Crotaphytus collaris, C.

reticulatus, Cophosaurus texanus, Holbrookia lacerata,

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The herpetofauna of Coahuila, Mexico

Table 9. SEMARNAT categorizations for herpetofaunal species in Coahuila, Mexico, arranged by families. Non-native species are

not included.

Number SEMARNAT Categorizations

Families of

. Special No status

species Endangered (P) | Threatened (A) protection (Pr) (NS)

(ee eee

Craugastoridae eee

Eleutherodactylidae a | Sy

Hylidae ee ee

Microhylidae ee

Ranidae Ee

Scaphiopodidae a. ac | a — as

Subtotals es

Ambystomatidae SSS Se

Plethodontidae SS a |

Subtotals ESS Sees

Totals Se ee eee

Anguidae _—EE ET Se

Crotaphytidae | Ss ee

Eublepharidae ——S SS

Phrynosomatidae

Scincidae (Se SS

Sphenomorphidae EE a ae

Teiidae ES ae

Xantusiidae a a

Subtotals a a kas)

Colubridae EE EEE

Dipsadidae a Ss |e ee

Elapidae aS a

Leptotyphlopidae ee ee

Natricidae SS ae

Viperidae ee

Subtotals ee (es

Emydidae ES ae

Kinosternidae =

Testudinidae

—)

a ha

a a ae

PS)

Phrynosoma_ orbiculare, Sceloporus ornatus, Uta The IUCN System. The system of conservation

stansburiana, Scincella silvicola, Coluber constrictor, categorization developed by the International Union for

Lampropeltis alterna, Masticophis flagellum, Pituophis — the Conservation of Nature (IUCN) is applied globally,

deppei, Tantilla atriceps, T. gracilis, Trimorphodon _ theoretically to all organisms, and consists of a set of

vilkinsonii, Nerodia erythrogaster, Thamnophis exsul, T; nine categories, divided into four general categories,

marcianus, T: proximus, Pseudemys gorzugi, Terrapene including: Extinct (Extinct and Extinct in the Wild),

coahuila, and Gopherus berlandieri. This group of 23. Threatened (Critically Endangered, Endangered, and

species includes a curious mixture of seven country and ~—- Vulnerable), Lower Risk (Near Threatened and Least

state endemic species and 16 relatively broadly ranging Concern), and other categories (Data Deficient and Not

non-endemic species, some with extensive ranges inthe — Evaluated). Those evaluations applying to the Coahuilan

United States. herpetofauna are shown in Table 7 and summarized in

Until such time as SEMARNAT provides conservation — Table 10.

evaluations for all Mexican herpetofaunal species, this Only 14 of 140 native species (10.0%) are judged to

system will continue to have only limited utility for | be Threatened. No species are considered to be Critically

conservation purposes. Endangered; seven are assessed as Endangered; and seven

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Lazcano et al.

Table 10. IUCN Red List categorizations for herpetofaunal families in Coahuila, Mexico. Non-native species are excluded. The shaded

columns to the left are the “threat categories,” and those to the right are the categories for which either too little information on conservation

status exists to allow the taxa to be placed in any IUCN category, or they have not been evaluated.

Families

| Subtotals | 19

| Ambystomatidae |

| Plethodontidae | 3

Number IUCN Red List categorizations

]

117

140

others as Vulnerable. The seven Endangered species

are: Gerrhonotus parvus*, Crotaphytus antiquus*”,

Sceloporus cyanostictus*, S. goldmani*, Uma exsul**,

Terrapene coahuila**, and Trachemys taylori**. Three

of these species are country endemics and four are state

endemics. The seven Vulnerable species (VU) are:

Eleutherodactylus longipes*, Aquiloeurycea scandens*,

Crotaphytus reticulatus, Sceloporus maculosus*, Storeria

hidalgoensis*, Trachemys gaigeae, and Gopherus

flavomarginatus*. Five of the seven VU species are

country endemics; the remainder are non-endemics. The

Lower Risk species amount to 102 species, constituting

72.9% of the native species in the state (Table 10). Of

these 102 species, eight are Near Threatened and 94

Amphib. Reptile Conserv.

are Least Concern. As has been demonstrated clearly in

other studies in the Mexican Conservation Series, the

allocation of such a large number of species to the Least

Concern category seems unjustified and the conservation

status of these 94 “Least Concern” species is examined in

greater detail in the following section.

Only a single Coahuilan species is considered to be

Data Deficient (Table 10): Kinosternon durangoense

(Table 7). This turtle is a country endemic that was

recognized as a distinct species in 2001 (Serb et al. 2001)

by being elevated from a subspecies of K. flavescens, but

very little information on its biology and conservation

status 1s available to date. Therefore, this appears to be a

sound assessment.

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The herpetofauna of Coahuila, Mexico

No. 13. Sceloporus samcolemani Smith and Hall, 1974. Coleman’s Bunch Grass Lizard is a Mexican endemic species that ranges

from “extreme southeastern Coahuila and southern central Nuevo Leon” (Watkins-Colwell et al. 1998: 675.2). This individual

was seen at Monterreal, in the municipality of Arteaga. Wilson et al. (2013a) gauged its EVS as 15, placing it in the lower portion

of the high vulnerability category. Its conservation status has been determined as Least Concern, but this lizard is not listed by

SEMARNAT. Photo by Michael S. Price.

No. 14. Uma exsul Schmidt and Bogert (1947). The Fringe-toed Sand Lizard is a Mexican endemic species occurring in extreme

southwestern and south-central Coahuila (Lemos-Espinal and Smith 2007). This individual was located at ca. 2 km SW from Ejido

Alejandria, in the municipality of San Pedro. Wilson et al. (2013a) assessed its EVS as 16, placing it in the middle portion of the

high vulnerability category. Its conservation status is evaluated as Endangered by IUCN and as endangered (P) by SEMARNAT.

Photo by Marco Antonio Bazdn-Tellez.

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Lazcano et al.

No. 15. Plestiodon dicei (Ruthven my Ge 1933). Bick: S Rho noel Skink j is a ars Meanie occurring from central

Nuevo Leon eastward to central and southern Tamaulipas (Feria-Ortiz et al. 2011). This individual was found in Monterreal, in the

municipality of Arteaga. Wilson et al. (2013a) calculated its EVS as 12, placing it in the upper portion of the medium vulnerability

category. Its conservation status has not been evaluated by IUCN and this species is not listed by SEMARNAT. Photo by Eli Garcia

Padilla.

No. 16. Bogertophis subocularis (Brown, 1901). The Trans-Pecos Rat Snake is distributed “in southern New Mexico, southwestern

Texas, and in northeastern Mexico, from Chihuahua through Coahuila and into Nuevo Leon and through Durango down to its border

with Zacatecas” (Lemos-Espinal et al. 2015: 318). This individual came from Cuatrociénegas, in the municipality of Cuatrociénegas

de Carranza. Wilson et al. (2013a) judged its EVS as 14, placing it at the lower edge of the high vulnerability category. Its conservation

status is evaluated as Least Concern by IUCN, but it is not listed by SEMARNAT. Photo by Michael S. Price.

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The herpetofauna of Coahuila, Mexico

Twenty-three species (16.4%) in the native Coahuilan

herpetofauna remain unevaluated (Table 10). The status

of these species is examined further using the EVS

system in the following section.

The EVS System. The EVS system, developed

originally for use in the conservation assessment of the

Honduran herpetofauna (Wilson and McCranie 2004),

has been applied subsequently to the entirety of Mexico

(Wilson et al. 2013a,b) and to Central America (Johnson

et al. 2015b), as well as to various states and groups

of states in Mexico (Alvarado-Diaz et al. 2013; Mata-

Silva et al. 2015; Johnson et al. 2015a; Teran-Juarez et

al. 2016; Woolrich-Pifia et al. 2016, 2017; Nevarez-de

los Reyes et al. 2016; Cruz-Saenz et al. 2017; Gonzalez-

Sanchez et al. 2017). Wilson et al. (2013a,b) described

the use of this system for the herpetofauna of Mexico;

and it is employed here to assess the conservation status

of the herpetofauna of Coahuila (see data in Table 7,

summarized in Table 11).

The total range of EVS values for the herpetofauna of

Coahuila spans the entire theoretical EVS range (3—20).

The most frequent values (more than 10 species) are nine

(10), 10 (10), 11 (12), 12 (17), 13 (17), 14 (14), 15 (13),

and 16 (12). These eight scores were applied to a total

of 104 species, or 74.3% of the total number of native

species. The average EVS value is 12.1 (1,693/140). When

allocated to the three summary categories of low (3-9),

medium (10-13), and high (14—20), the species counts for

these categories are 32, 56, and 52, respectively.

The lowest EVS value of 3 was applied to three

species of anurans (Rhinella horribilis, Smilisca baudinii,

and Scaphiopus couchii), which are geographically and

ecologically widespread and have the most widespread

reproductive mode (eggs and tadpoles in still water).

At the other extreme, one species (the trionychid turtle

Apalone atra) was provided a value of 20, because of

its narrow geographic and ecological distribution and its

high level of human persecution.

Comparing the results of the IUCN categorization

with those of the EVS system in Table 12, indicates that

14 of the 52 high vulnerability species (26.9%) are judged

to occupy one of two IUCN threat categories (EN or VU,

note that no species are allocated to the CR category).

Seven species (five lizards and two turtles) are evaluated

as EN, and seven as VU (one anuran, one salamander,

two lizards, one snake, and two turtles). These 14

species comprise 10.0% of the 140 native herpetofaunal

species in Coahuila. At the opposite extreme, the 32

low vulnerability species comprise 35.1% of the 93

LC species. As in other Mexican Conservation Series

studies, the results of the applications of the IUCN and

EVS systems do not complement one another.

Twenty-four species remain unassessed using the

IUCN system (allocated to the NE category in Table 7).

Four of these species are state endemics (Gerrhonotus

mcecoyi, Sceloporus gadsdeni, Scincella_ kikaapoa,

Amphib. Reptile Conserv.

and Apalone atra), five are country endemics (Barisia

imbricata, Holbrookia approximans, Plestiodon dicei,

Lampropeltis leonis, and Crotalus morulus), and the

remainder are non-endemics. The range of EVS values

for these 24 species is 6-20, which places some of them

into each of the three summary categories (Table 13).

Four have low EVS scores, ten have medium scores,

and ten have high scores. Until such time as IUCN

evaluations are available for these species, we suggest

that the high EVS species should be placed in one of

the three threat categories, perhaps as follows: CR—

Gerrhonotus mccoyi, Sceloporus gadsdeni, Scincella

kikaapoa, Lampropeltis leonis, Crotalus morulus,

Apalone atra; EN—Barisia imbricata, Holbrookia

approximans, VU—Aspidoscelis marmorata, Salvadora

deserticola. We also suggest that the species with EVS of

12 or 13 be placed in the NT category. The remainder of

the species with EVS of 6-11 can be allocated to the LC

category (Table 13).

As with other studies in the Mexican Conservation

Series, this study found that a significantly large number

of the Coahuilan herpetofauna members have been

allocated by the IUCN to the Least Concern category.

The number of such species amounts to 93 (66.4% of the

total of 140 species). Given that almost seven of every

ten herpetofaunal species in Coahuila is judged Least

Concern, it might appear that the state herpetofauna is in

relatively good shape with respect to conservation status.

In order to ascertain whether such an optimistic view is

the case, the 93 species in Table 14 were placed along

with the calculations for their respective EVS values.

Although one might expect that the LC species would

most likely be non-endemic to Mexico, this analysis

found that 12 are country endemics, including one

salamander, nine lizards, and two snakes, and one lizard

is a state endemic (Table 14). The range of EVS values

for these 93 species is 3—18, or only slightly less than the

entire theoretical range for EVS (3-20). The allocation of

the EVS values for the 93 species into the three summary

categories demonstrates the following: low (3—9)—27;

medium (10-13)—45; and high (14—-20)—22. Based

on these allocations, we suggest that a more realistic

assessment would place the 22 high vulnerability

species in one of the three IUCN threat categories, as

follows: CR (Gerrhonotus lugoi, Thamnophis exsul,

and Gopherus berlandieri),; EN (Coleonyx reticulatus,

Sceloporus cautus, S. couchii, S. parvus, S. samcolemani,

Xantusia extorris, Lampropeltis mexicana, Pantherophis

bairdi, and Apalone spinifera); and VU (Coleonyx brevis,

Cophosaurus texanus, Sceloporus minor, Aspidoscelis

inornata, A. tesselata, Bogertophis subocularis,

Lampropeltis alterna, Pituophis deppei, Agkistrodon

laticinctus, and Crotalus pricei). We also suggest that

the 43 medium vulnerability species probably should

be placed in the NT category, and that the 27 low

vulnerability species could remain in the LC category.

October 2019 | Volume 13 | Number 2 | e189

Lazcano et al.

PEE EEDELE DEEL EELLTERED EDEL EL [Ee

PAPEETE EEDA EEDE EERE EDEL

PEEELED EDEL EESTELELL

CUEEEE TELE D LED PELL

SD EDO:

EDODOUD CHUE

wilt

aie

ale

OOOOH OEE

OOOUODOND

OOOUOUOUOUEDOND

DUODODODODODODOODOROD

m= | O ois N

a) =

“recePeeeree TTT

rr)

ars

i=)

i—|

aie

Number

Species

Families

Leptotyphlopidae

Category Totals

Natricidae

Sphenomorphidae

Viperidae

Ambystomatidae

Plethodontidae

Subtotals

Kinosternidae

Testudinidae

Trionychidae

Subtotals

Totals

Sum Totals

Scincidae

Xantustidae

Subtotals

Colubridae

Dipsadidae

Elapidae

Subtotals

Emydidae

Table 11. Environmental Vulnerability Scores (EVS) for herpetofaunal species in Coahuila, Mexico, arranged by family. Shaded area to the left encompasses low vulnerability scores, and the

Subtotals

one to the right high vulnerability scores. Non-native species are excluded.

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The herpetofauna of Coahuila, Mexico

Table 12. Comparison of Environmental Vulnerability Scores (EVS) and applicable IUCN categorizations for members of the

herpetofauna of Coahuila, Mexico. Non-native species are excluded. No species are allocated to the CR IUCN categories. Shaded

area at the top encompasses low vulnerability category scores, and the one at the bottom high vulnerability category scores.

IUCN Categories

Mi Near

Vulnerable ‘Tivventétied

Endangered

—

]

Table 13. Environmental Vulnerability Scores (EVS) for members of the herpetofauna of Coahuila, Mexico, currently not evaluated

(NE) by the IUCN. Non-native taxa are not included. * = country endemic species; ** = state endemic species.

Environmental Vulnerability Score

Geographic Ecological Heprouucliye

Ps OE ST Ae Mode/Degree of Total Score

Distribution Distribution ‘

Persecution

Acris blanchardi

Barisia ciliaris*

Gerrhonotus mccoyi**

Holbrookia approximans*

Sceloporus cowlesi

Sceloporus cyanogenys

Sceloporus gadsdeni**

Sceloporus marmoratus

Plestiodon dicei*

Scincella kikaapoa**

Aspidoscelis marmorata

Lampropeltis annulata

Lampropeltis gentilis

Lampropeltis leonis*

Lampropeltis splendida

Salvadora deserticola

Sonora episcopa

Heterodon kennerlyi

Hypsiglena jani

Leptodeira septentrionalis

Rena segrega

Crotalus morulus

Crotalus ornatus

Apalone atra**

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Lazcano et al.

Table 14. Environmental Vulnerability Scores (EVS) for members of the herpetofauna of Coahuila, Mexico, assigned to the IUCN

Least Concern category. Non-native taxa are not included. * = country endemic species; ** = state endemic species.

Environmental Vulnerability Score

: : Reproductive

ae Geographic Eeoloeicg! Mode/Degree of Total Score

Distribution Distribution :

Persecution

5 5

Anaxyrus cognatus 1

Anaxyrus debilis

Anaxyrus punctatus

—

Anaxyrus Speciosus

—

Anaxyrus woodhousii

Incilius nebulifer

Rhinella horribilis

Craugastor augusti

—

Eleutherodactylus cystignathoides

—

Eleutherodactylus guttilatus

Dryophytes arenicolor

Smilisca baudinii

Gastrophryne olivacea

Lithobates berlandieri

Scaphiopus couchii

Spea multiplicata

—

is)

Ambystoma velasci*

—

ies)

Gerrhonotus infernalis

—

Gerrhonotus lugoi**

—

ies)

Crotaphytus collaris

—

Crotaphytus wislizenii

—

Coleonyx brevis

—

Coleonyx reticulatus

—

™~

Cophosaurus texanus

Phrynosoma cornutum

Phrynosoma modestum

—

Phrynosoma orbiculare*

—

Sceloporus cautus*

—

Sceloporus couchii*

Sceloporus grammicus

Sceloporus magister

—

Sceloporus merriami

—_

—

Sceloporus minor*

—

ies)

Sceloporus olivaceus

—

Sceloporus parvus*

—

Sceloporus poinsettii

—

Sceloporus samcolemani*

—

Sceloporus spinosus*

—

Urosaurus ornatus

Uta stansburiana

Plestiodon obsoletus

—

Plestiodon tetragrammus

—

ies)

Scincella lateralis

—

Scincella silvicola*

Aspidoscelis gularis

—

Aspidoscelis inornata

—

™~

Aspidoscelis tesselata

—

Xantusia extorris*

Arizona elegans

—

Bogertophis subocularis

—

os)

Coluber constrictor

TEER THETEPRRER HHT {tHE

HHA RTH TRE

TREE

TEPEEPPTTCPPEEE RTP {TTP

Amphib. Reptile Conserv. 73 October 2019 | Volume 13 | Number 2 | e189

The herpetofauna of Coahuila, Mexico

Table 14 (continued). Environmental Vulnerability Scores (EVS) for members of the herpetofauna of Coahuila, Mexico, assigned

to the IUCN Least Concern category. Non-native taxa are not included. * = country endemic species; ** = state endemic species.

Geographic

Distribution

Diadophis punctatus

Nerodia erythrogaster

Tantilla nigriceps

Tantilla wilcoxi

Nerodia rhombifer

Trimorphodon vilkinsonii

Apalone spinifera

Relative Herpetofaunal Priority

Johnson et al. (2015a) developed the concept of Relative

Herpetofaunal Priority (RHP), a simple means for

measuring the relative importance of the herpetofaunal

species found in any geographic entity (e.g., a state or

a physiographic region). Determining the RHP involves

the use of two methods, i.e., (1) calculating the proportion

Amphib. Reptile Conserv.

Environmental Vulnerability Score

Reproductive

Mode/Degree of

Persecution

Ecological

Distribution

Total Score

of state and country endemics relative to the entire

physiographic regional herpetofauna, and (2) computing

the absolute number of EVS high category species in

each physiographic regional herpetofauna.

Here, two tables have been constructed to ascertain

the RHP values for the Coahuilan herpetofauna, one for

the endemicity values (Table 15) and the other for the

high category EVS values (Table 16). The data in Table

74 October 2019 | Volume 13 | Number 2 | e189

Lazcano et al.

Table 15. Numbers of herpetofaunal species in four distributional categories among the 10 physiographic provinces of Coahuila,

Mexico. Rank determined by adding state and country endemics.

Physiographic Province Non- Country State Non-

ae endemics endemics endemics natives

5 2 2

Llanuras de Coahuila y Nuevo Leon

]

ir ee

= a a a a

rhe ee eS ae

15 demonstrate that the highest amount of endemicity is

found in the Gran Sierra Plegada, only a small portion

of which is located in Coahuila (Fig. 1); thus, this region

occupies rank number 1 using this measure. Of the 51

species recorded in this region, 18 are country endemics

(35.3%). Nevarez-de los Reyes et al. (2016) found a

similar situation prevailing in the neighboring state

of Nuevo Leon, in which 33 of 87 species (37.9%) in

this same region consist of country and state endemics.

Rank number 2 is occupied by the Sierras Transversales

located along the southern border of Coahuila, in which

13 of 46 species (28.3%) are country endemics. The

remaining eight physiographic regions occupy ranks 3

through 7, given that several of these regions share the

same rank with other regions (Table 15). One region

(Sierras y Llanuras Coahuilenses) occupies rank 3, and

contains eight country and state endemics. Only one

region occupies rank 4, which is the Bolson de Mapimi,

with five country endemics and two state endemics. One

region (Pliegues Saltillo Parras) lies at rank 5, with six

country endemics. Two regions (Laguna de Mayran and

Sierra de la Paila) occupy rank 6, with two endemics

each. Finally, there are three regions (Llanuras y Sierras

Volcanicas, Serranias del Burro, and Llanuras de

Coahuila y Nuevo Leon) that occupy rank 7, each with a

single endemic species.

The numbers of species for each of the 10

physiographic regions are placed into the three EVS

—tRee

CO | LW

categories (low, medium, and high) in Table 16.

These data indicate that the greatest number of high

vulnerability species (22 of 88, 25.0%) is found in the

Sierras y Llanuras Coahuilenses in the central portion of

the state (Fig. 1), thus this region occupies rank number

1. The next highest number (17 of 50 species, or 34.0%)

is found in the Gran Sierra Plegada, which is located in

the southeastern corner of the state (Fig. 1) and occupies

rank number 2. The 3 rank is occupied by the Sierras

Transversales, with 14 high-vulnerability species out

of 46 (30.4%). The numbers of high EVS species in the

remaining seven regions range from seven to 13.

The rankings obtained by using these two RHP

methods are not identical, but at least the regions

occupying ranks | through 3 are the same three regions 1n

each case, as follows (endemic species ranking followed

by high vulnerability ranking):

Gran Sierra Plegada (1, 2)

Sierras Transversales (2, 3)

Sierras y Llanuras Coahuilenses (3, 1)

The results of this RHP analysis clearly show that

the most important region in the state is the Gran Sierra

Plegada, occupying rank 1 for endemic species and

rank 2 for high vulnerability species. This finding is

interesting, given the small amount of this physiographic

region situated in Coahuila (2,178 km”, as noted above),

Table 16. Number of herpetofaunal species in the three EVS categories among the 10 physiographic regions of Coahuila, Mexico.

Rank determined by the relative number of high EVS species. Non-native species are excluded.

Rank

Order

Amphib. Reptile Conserv.

October 2019 | Volume 13 | Number 2 | e189

The herpetofauna of Coahuila, Mexico

a . ’ Cs . = 7 ees lee” a

r ee ee . a eh ome aii ae oe

~ tlP ra ~~

mt, xs ae any ad . } Ros < “ue ae ee ; 4 he

et Laces ee a . PD ee Sou ment

-¥'

No. 17. Rhinocheilus lecontei Baird and Girard, 1853. The Long-nosed Snake occurs from “California to Kansas, excluding much

of the Great Basin and the Rocky Mountains, southward to Baja California and Nayarit and, east of the Sierra Madre [Occidental],

to the southern limited of the Chihuahua Desert” (Lemos-Espinal et al. 2015: 371). This individual was located at ca. 4 km east of

Nava, in the municipality of Nava. Wilson et al. (2013a) assessed its EVS as 8, placing it in the upper portion of the low vulnerability

category. Its conservation status is judged as Least Concern by IUCN, but it is not listed by SEMARNAT. Photo by Marco Antonio

Bazdn-Tellez.

a ee aes Se = -«< .a 4 ™

No. 18. Zantilla atriceps (Gunther, 1895). The Mexican Black-headed Snake is distributed from “extreme southern Texas southward

through central Coahuila to extreme northeastern Durango and northern Tamaulipas and San Luis Potosi” (Lemos-Espinal et al.

2015: 382). This individual came from Rancho La Boca, on the border of the municipalities of Bustamante and Mina. Wilson et al.

(2013a) calculated its EVS as 11, placing it in the lower portion of the medium vulnerability category. Its conservation status has

been considered as Least Concern by IUCN, and as a threatened species (A) by SEMARNAT. Photo by Michael S. Price.

Amphib. Reptile Conserv. 76 October 2019 | Volume 13 | Number 2 | e189

Lazcano et al.

No. 19. Micrurus tener (Baird and Girard, 1853). The Texas Coralsnake occurs “from the Mississippi River westward into Texas,

in the United States, and in Mexico, from Tamaulipas south to Veracruz...” (Lemos-Espinal and Dixon 2013: 240). This individual

came from ca. 16 km east of Nava, in the municipality of Nava. Wilson et al. (2013a) calculated its EVS as 11, placing it in the

middle of the medium vulnerability category. Its conservation status has been determined as Least Concern by IUCN, and this

species is not listed by SEMARNAT. Photo by Marco Antonio Bazan-Tellez.

(ied a

- Wide ie

es s

a -* .

~~ “< 8, — =

_ —- P

F ~~ 2S

- A,

. — na — - 2 —— = = =

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. -—

- ~~

No. 20. Crotalus atrox Baird and Girard, 1853. The Western Diamondback Rattlesnake occupies “much of the southwestern USA

south to northeastern Baja California, Sonora and northern Sinaloa, and east of the Sierra Madre Occidental from Chihuahua east to

Tamaulipas and south to Hidalgo and Veracruz” (Rorabaugh and Lemos-Espinal 2016: 573). This individual was found at ca. 1 km

NW from Ejido Mieleras, in the municipality of Viesca. Wilson et al. (2013a) determined its EVS as 9, placing it at the upper limit

of the low vulnerability category. Its conservation status is judged as Least Concern by IUCN and as a species of special protection

(Pr) by SEMARNAT. Photo by Marco Antonio Bazdn-Tellez.

Amphib. Reptile Conserv. 77 October 2019 | Volume 13 | Number 2 | e189

The herpetofauna of Coahuila, Mexico

although it has considerably more area outside the

state. The two other regions indicated above (Sierras

Transversales and Sierras y Llanuras Coahuilenses) are

considerably larger. The Gran Sierra Plegada supports 18

country endemics (Table 15) and 17 high vulnerability

species (Table 16). The Sierras Transversales contains 13

country endemics (Table 15) and 14 high vulnerability

species (Table 16); the respective figures for the Sierras

y Llanuras Coahuilenses are eight endemic species and

22 high vulnerability species. The level of protection

provided for these various species is indicated in the

following section on protected areas.

Protected Areas in Coahuila

A system of formally protected areas is integral to any

effort to protect any portion of the planetary biota from

the principal anthropogenic impacts brought about by

habitat degradation and destruction. Ostensibly, such a

system should incorporate as much of the environmental

diversity that exists within the target region (in this

case, the state of Coahuila) and as large of a portion of

that patrimony as is feasible within existing economic

confines. In an effort to determine how these concerns

have been addressed in Coahuila, information on

the protected areas in the state has been collated and

presented in Table 17.

The data in Table 17 indicate that of the 19 protected

areas listed, eight are federal reserves, four are federal/

private reserves, three are state reserves, three are state/

private reserves, and one is a municipal reserve. The

eight areas administered at the federal level include one

biosphere reserve, one national park, three floral and

faunal protection areas, two resource protection areas,

and one national monument.

These 19 areas in Coahuila have been established over

the 100 years from 1915 to 2015. Thirteen of these 19

areas have been in existence only since the turn of the

century or thereafter, while three were established in the

decade of the 1990s, two in the decade of the 1940s, and

one in 1915. Thus, it remains to be seen in the analysis

of the remainder of the data in Table 17 exactly what has

been accomplished in these areas, especially since 68.4%

of them (13/19) have existed for fewer than 17 years.

The areas of coverage of these protected areas range

broadly from as low as 38 ha to as high as 1,519,385 ha.

Interestingly, the largest of the 19 areas is the Area de

Proteccion de Recursos Naturales Cuenca Alimentadora

del Distrito Nacional de Riego 04 Don Martin,

established in 1915, which 1s the one with the longest

existence. The total coverage of these areas is 2,717,443

ha or approximately 27,174 km?, which is 17.9% of the

area of Coahuila. Portions of these 19 areas are located in

23 of the 38 municipalities (60.5%).

This system of protected areas in the state contains

representatives of eight of the ten physiographic regions,

including the Bols6n de Mapimi (one of 20 areas),

Amphib. Reptile Conserv.

Llanuras y Sierras Volcanicas (two), Laguna de Mayran

(one), Sierras y Llanuras Coahuilenses (eight), Serranias

del Burro (two), Sierra Transversales (four), Gran Sierra

Plegada (four), and Llanuras de Coahuila y Nuevo

Leon (two). The two regions with no representation are

the Sierra de la Paila and the Pliegues Saltillo Parras.

It is no doubt fortuitous that the three physiographic

regions with the greatest amount of representation in

the protected areas system are the regions having the

great herpetofaunal significance, 1.e., the Gran Sierra

Plegada, Sierras Transversales, and Sierras y Llanuras

Coahuilenses. Unfortunately, however, of the protected

areas for which such information is available, all are

known to be occupied to some extent by landowners.

Management plans are known to be available for only

five of the federal protected areas, which points out the

great need for completion of such plans for the remainder

of the areas. Even less well represented are herpetofaunal

surveys, of which there are only three completed and

three in the process of completion. Completion of the

remainder of the surveys needs to be undertaken as soon

as possible.

In general, systems of protected areas are established

without consideration of the conservation needs of the

herpetofauna, so it 1s not surprising that this need is

still outstanding in Coahuila. The good news is that the

physiographic regions that are herpetofaunally most

important, based on our RHP analyses, are those best

supplied with protected areas. The bad news is that sizable

proportions of those areas are occupied by landowners,

have no management plans designed, and have been

subject to no herpetofaunal surveys. Thus, at present,

there is no chance of answering any of the more pressing

questions about the state of population sustainability

of the component species of Coahuila’s herpetofauna.

So, redressing these inadequacies in the design and

implementation of the protected areas system has to be

undertaken so that the ability of the current system of

protected areas to provide for perpetual protection of the

state’s herpetofauna can be assessed and strengthened.

Here, the expected herpetofaunal content of the 19

protected areas in Coahuila is catalogued by compiling

the species known to occupy the physiographic regions

represented in each of these regions. We employed this

means since too few herpetofaunal surveys have been

undertaken in these areas to date. Thus, ground-based

field surveys will be necessary to provide the empirical

data required to substantiate the actual herpetofaunal

content of the state’s protected areas. Using this approach,

the results are shown in Table 18, and summarized in

Table 19.

Of the 143 species recorded in Coahuila, 120 (83.9%)

are expected to be found in one or more of the 19 protected

areas (Table 19). The number of species expected to

occur in each area ranges from three in the Parque

Estatal Bosque Urbano Ejército Mexicano to 84 in the

Area de Proteccién de Flora y Fauna Cuatro Ciénegas.

October 2019 | Volume 13 | Number 2 | e189

Lazcano et al.

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Amphib. Reptile Conserv.

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The herpetofauna of Coahuila

October 2019 | Volume 13 | Number 2 | e189

Amphib. Reptile Conserv.

Lazcano et al.

Table 18. Distribution of herpetofaunal species in Natural Protected Areas of Coahuila, Mexico, based on estimated inclusions. Abbreviations are as

follows: * = species endemic to Mexico; ** = species endemic to Coahuila; and *** = non-native species.

Natural Protected Area

Taxa ‘ : :

[Anora G@specie)

[Butonitae 7 specie

Angas cognats

[Amcgras debs

Angra puncahs

[Ancyras spcios

[énciyas woodkous

nce neler

Rhinela horrible

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Eleutherodactylidae (2 species)

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Eleuthera

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Hylidae (1 species)

Dryophytes arenicolor

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PRankse @ pects)

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Seaphiopodine @ pecs)

Seaphioms cowh

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Gerrononus mecoy™®

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Amphib. Reptile Conserv. 81 October 2019 | Volume 13 | Number 2 | e189

The herpetofauna of Coahuila, Mexico

Table 18 (continued). Distribution of herpetofaunal species in Natural Protected Areas of Coahuila, Mexico, based on estimated inclusions.

Abbreviations are as follows: * = species endemic to Mexico; ** = species endemic to Coahuila; and *** = non-native species.

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October 2019 | Volume 13 | Number 2 | e189

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Lazcano et al.

Table 18 (continued). Distribution of herpetofaunal species in Natural Protected Areas of Coahuila, Mexico, based on estimated inclusions.

Abbreviations are as follows: *

= non-native species.

‘and ***

= species endemic to Coahuila

Natural Protected Area

oR OK

= species endemic to Mexico;

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ona elegans

Colubridae (26 species)

Ariz

Coluber constrictor

Gyalopion canum

Lampropeltis leonis*

Lampropeltis splendida

Masticophis flagellum

Opheodrys aestivus

Tantilla atriceps

Tantilla gracilis

Tantilla hobartsmithi

Tantilla nigriceps

Masticophis taeniatus

Rhinocheilus lecontei

Scincella lateralis

Teiidae (4 species)

Aspidoscelis gularis

Aspidoscelis inornata

Aspidoscelis marmorata

Aspidoscelis tesselata

Xantusiidae (1 species)

Xantusia extorris*

Bogertophis subocularis

Drymarchon melanurus

Lampropeltis alterna

Lampropeltis annulata

Masticophis schotti

Pantherophis bairdi

Pantherophis emoryi

Pituophis catenifer

Pituophis deppei*

Salvadora deserticola

Salvadora grahamiae

Sonora episcopa

October 2019 | Volume 13 | Number 2 | e189

Amphib. Reptile Conserv.

The herpetofauna of Coahuila, Mexico

Table 18 (continued). Distribution of herpetofaunal species in Natural Protected Areas of Coahuila, Mexico, based on estimated inclusions.

Abbreviations are as follows: * = species endemic to Mexico; ** = species endemic to Coahuila; and *** = non-native species.

Natural Protected Area

Dipsadidae (4 species)

Diadophis punctatus

Heterodon kennerlyi

Hypsiglena jani

Leptodeira septentrionalis

Elapidae (1 species)

Micrurus tener

Leptotyphlopidae (3 species)

Rena dissecta

Rena dulcis

Rena segrega

Natricidae (7 species)

Nerodia erythrogaster

Nerodia rhombifer

Storeria hidalgoensis*

Thamnophis cyrtopsis

Thamnophis exsul*

Thamnophis marcianus

Thamnophis proximus

Viperidae (9 species)

Agkistrodon contortrix

Crotalus atrox

Crotalus lepidus

Crotalus molossus

Crotalus morulus*

Crotalus ornatus

Crotalus pricei

Crotalus scutulatus

Sistrurus tergeminus

Testudines (11 species)

Emydidae (5 species)

Pseudemys gorzugi

Terrapene coahuila**

Trachemys gaigeae

Trachemys scripta***

Amphib. Reptile Conserv.

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84 October 2019 | Volume 13 | Number 2 | e189

Lazcano et al.

Table 18 (continued). Distribution of herpetofaunal species in Natural Protected Areas of Coahuila, Mexico, based on estimated inclusions.

Abbreviations are as follows: * = species endemic to Mexico; ** = species endemic to Coahuila; and *** = non-native species.

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Trionychidae (2 species)

Apalone atra**

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Of the 120 species included in Table 18, 26 (21.0%) are

endemic species, including seven (5.8%) state endemics

(Gerrhonotus mccoyi, Crotaphytus antiquus, Uma exsul,

Scincella kikaapoa, Terrapene coahuila, Trachemys

taylori, and Apalone atra). Ninety-one of the 120 species

(75.8%) are non-endemics, and three (2.5%) are non-

natives (all of the three that occur in the state). Naturally,

it is not desirable to have the non-native species within

protected areas, but, fortunately, only one of the three

(Hemidactylus turcicus) 1s expected to be found in more

than three of the areas (and it is expected in all of them).

Of the 23 species that are not expected to be found

within the 19 existing protected areas, 12 are country

endemics:

Eleutherodactylus longipes

Rheohyla miotympanum

Ambystoma velasci

Aquiloeurycea galeanae

Aquiloeurycea scandens

Chiropterotriton priscus

Barisia imbricata

Phrynosoma orbiculare

Sceloporus cyanostictus

Sceloporus goldmani

Sceloporus minor

Scincella silvicola

One of the 23 species (Sceloporus gadsdeni) is a state

endemic and 10 are non-endemics (Acris blanchardi,

Smilisca baudinii, Holbrookia lacerata, Sceloporus

Amphib. Reptile Conserv.

marmoratus, Lampropeltis gentilis, Tantilla cucullata,

Trimorphodon vilkinsonii, Crotalus viridis, Terrapene

ornata, and Kinosternon hirtipes).

The principal herpetofaunal conservation goal for

Coahuila, at this point, is to conduct herpetofaunal

surveys in all currently-established conservation areas to

determine which species are now actually represented.

Based on this analysis, we predict that relatively few

Species are expected to be absent from all of these 19

areas, So a subsidiary goal is to ascertain whether this 1s

the case and, if so, what other areas could be established

to contain them.

Conclusions and Recommendations

Conclusions

A. At the present time, the herpetofauna of Coahuila

comprises 143 species, including 20 anurans, four

salamanders, 106 squamates, and 13 turtles; three species

are non-natives.

B. The number of herpetofaunal species distributed

among the 10 physiographic regions we recognize in

Coahuila varies from 38 in the Laguna de Mayran region

to 91 in the Sierras y Llanuras Coahuilenses.

C. The level of endemism of the herpetofauna of

Coahuila is relatively low. Of the 143 species recorded

from the state, 40 are endemic to Mexico, including nine

limited to Coahuila. Thus, the percentage of endemism

October 2019 | Volume 13 | Number 2 | e189

The herpetofauna of Coahuila, Mexico

Table 19. Summary of the distributional status of herpetofaunal species in protected areas in Coahuila, Mexico. Totals = total

number of species recorded in all of the listed protected areas.

Protected Areas Number

Los Novillos

Cuenca Alimentadora del Distrito

Nacional de Riego 04 Don Martin

Distrito Nacional de Riego 026 Bajo

Rio San Juan.

Rancho Media Luna

Rancho La Puerta

Tierra Silvestre Cafion del Diablo

is 28.6%. The 40 endemic species amount to 4.9% of the

811 endemic species in Mexico.

D. The distributional status of the Coahuilan herpetofauna

is as follows (in order of the size of the categories): non-

endemics (100, 69.9%); country endemics (31, 21.7%);

state endemics (nine, 6.3%); and non-natives (three,

2.1%).

E. The principal environmental threats in Coahuila are

urban development, industrial pollution, deforestation

for agricultural and ranching purposes, road effects,

mining and energy projects, natural gas fracking, wind

turbines, elimination due to cultural beliefs and practices,

collecting and commercial trade, and forest fires.

F. The SEMARNAT, IUCN, and EVS systems were used

to evaluate the conservation status of the herpetofauna

of Coahuila. As demonstrated in previous MCS studies,

the SEMARNAT system proved to be of little value,

inasmuch as only 35.7% of the native herpetofauna

has been evaluated to date. Of these 50 species, four

are placed in the endangered category (P), 23 in the

threatened category (A), and 23 in the special protection

category (Pr).

G. The IUCN system was also applied to assess the native

Coahuilan herpetofauna, and the results (by category

Amphib. Reptile Conserv.

Distributional Status

(NE) Endemic (CE) (SE) (NN)

|RioBravodelNorte | 76 | 8 3

| MaderasdelCarmen | 79TH

and proportion) are: CR (0 of 140 species; 0%); EN (7;

5.0%); VU (7; 5.0%); NT (8; 5.7%); LC (94; 67.1%); DD

(1; 0.7%); and NE (23; 16.4%).

H. In addition, the EVS system was applied to the 140

native Coahuilan species. It placed them in the low,

medium, and high vulnerability categories, and the

values increased from low (33; 23.6%) to medium (55;

39.3%) and then slightly decreased in the high category

(52; 37.1%).

I. The IUCN and EVS conservation status allocations

ascertained that only 26.9% of the EVS high vulnerability

species have been placed in two of the three IUCN

threat categories (EN and VU; while no species are

allocated to the CR category) and only 35.1% of the EVS

low vulnerability species have been placed in the LC

category. As such, the results of the application of these

two systems do not correspond well to one another.

J. An analysis of the conservation status of the 118

species placed in the IUCN DD, NE, and LC categories

demonstrates that many of them have been evaluated

inappropriately compared to their respective EVS values.

We opine that these species need to be reassessed to

better reflect their prospects for survival.

K. The Relative Herpetofaunal Priority (RHP) measure

October 2019 | Volume 13 | Number 2 | e189

Lazcano et al.

Fig. 23. Forest Fires. The scene after a forest fire in the vicinity of Arteaga, in the municipality of Arteaga. Photo by Manuel

Nevarez de los Reyes.

was applied to establish the conservation significance

of the ten regional herpetofaunas in Coahuila, which

indicates that the most significant herpetofauna 1s that

of the Gran Sierra Plegada, as it contains the greatest

number of country endemics and the second greatest

number of high vulnerability species. The other nine

physiographic regions are arranged in decreasing order

of significance on the basis of their number of endemic

species, as follows: Sierra Transversales; Sierras y

Llanuras Coahuilenses; Bolson de Mapimi; Pliegues

Saltillo Parras; Laguna de Mayran and Sierra de la Paila;

Llanuras y Sierras Volcanicas, Serranias del Burro, and

Llanuras de Coahuila y Nuevo Leon. On the basis of their

numbers of high vulnerability species, the ranking is as

follows: Sierras y Llanuras Coahuilenses; Gran Sierra

Plegada; Sierras Transversales; Pliegues Saltillo Parras;

Bolson de Mapimi; Llanuras y Sierras Volcanicas and

Serrania del Burro; Laguna de Mayran, Sierra de la Paila,

and Llanuras de Coahuila y Nuevo Leon.

L. Nineteen protected areas have been established in

Coahuila; eight federal reserves, four federal/private

reserves, three state reserves, three state/private reserves,

and one municipal reserve. The representation of these

19 areas among the ten physiographic areas is weighted

in favor of the Sierras y Llanuras Coahuilenses (eight

areas), which ranked 3“ in endemic species and 1* in high

vulnerability species. The Gran Sierra Plegada is the next

best represented (in four areas). Unfortunately, all of the

19 protected areas for which information is available are

occupied by landowners. In addition, few areas have the

Amphib. Reptile Conserv.

benefit of herpetofaunal surveys or management plans.

M. Our analyses predict that 120 of 143 total species are

expected to be found in the 19 protected areas (83.9%).

These 120 species include 91 non-endemics, 19 country

endemics, seven state endemics, and three non-natives.

The non-native species should not be included the

protected areas system.

N. Future conservation efforts should be directed

toward conducting thorough herpetofaunal surveys in

all components of the protected areas system, as well as

determining what additional areas might be required to

provide protection for all of Coahuila’s herpetofaunal

species.

Recommendations

A. Our principal interest in writing this paper has

been to assess the conservation status of the 140

native species presently recorded from the state of

Coahuila, and to suggest what steps need to be taken

to protect all of these species over the long term.

We have undertaken this assessment using the EVS

methodology, as we have in the previous entries in the

Mexican Conservation Series, which demonstrated

that 33 species are allocated to the low vulnerability

category, 55 to the medium vulnerability category, and

52 to the high vulnerability category. We also employed

the Relative Herpetofaunal Priority methodology to

determine which of the physiographic regions in the

October 2019 | Volume 13 | Number 2 | e189

The herpetofauna of Coahuila, Mexico

No. 21. eis Tees (Kennicott, a The ae Rattlesnake is distributed “in the United States in eeouihesstern Arizona,

southern New Mexico, and western Texas” and “in Mexico...in Sonora, Chihuahua, Durango, Sinaloa, Nayarit, Jalisco, Zacatecas,

Aguascalientes, Coahuila, Nuevo Leon, and San Luis Potosi” (Prival and Porter 2016: 444). This individual came from Jimenez,

in the municipality of Jimenez. Wilson et al. (2013a) evaluated its EVS as 12, placing it in the upper portion of the medium

vulnerability category. Its conservation status 1s considered as Least Concern by IUCN and as a species of special protection (Pr) by

SEMARNAT. Photo by Michael S. Price.

NG IAI

No. 22. eee pricei Van Seine: 1895. “The Twin- etn es ranges “from ee ean in the United

States (Chiricahua, Huachuca, Pinaleno, Dos Cabezas, and Santa Rita mountains) southward in Mexico through the Sierra Madre

Occidental to northeastern Sonora, western Chihuahua, and Durango, and in the Sierra Madre Oriental of southeastern Coahuila,

southern Nuevo Leon, southwestern Tamaulipas, and north-central San Luis Potosi, and in Aguascalientes” (Hammerson et al.

2007). This individual was found at Monterreal, in the municipality of Arteaga. Wilson et al. (2013a) assessed its EVS as 14, placing

it at the lower limit of the high vulnerability category. Its conservation status is judged as Least Concern by IUCN and as a species

of special protection (Pr) by SEMARNAT. Photo by Eli Garcia Padilla.

Amphib. Reptile Conserv. 88 October 2019 | Volume 13 | Number 2 | e189

Lazcano et al.

No. 23, Sa eels (Say, 1823). The weston WMastesaea occurs in ihe USA (Texas, Otisnona) and Wiexen (central

and northeastern Coahuila, southern Nuevo Leon; possibly in Tamaulipas, northern Chihuahua, and northeastern Sonora). This

individual came from 6 km south of La Piedra Parada, in the municipality of Guerrero. The EVS value of this rattlesnake is 13. Its

conservation status has not been assessed by IUCN and it is not listed by SEMARNAT. Photo by Manuel Nevarez de los Reyes.

No. 24. Terrapene coahuila Schmidt a Owens, 1944. The Cent BOs Turtle i isa Mena Son species restricted to “the

Cuatro Ciénegas Bolson of Coahuila” (Lemos-Espinal et al. 2015: 122). This individual was located at Cuatrociénegas in the

municipality of Cuatrociénegas de Carranza. Wilson et al. (2013a) calculated its EVS as 19, placing it in the upper portion of the

high vulnerability category. Its conservation status is determined as Endangered by IUCN and as threatened (A) by SEMARNAT.

Photo by Michael S. Price.

Amphib. Reptile Conserv. 89 October 2019 | Volume 13 | Number 2 | e189

The herpetofauna of Coahuila, Mexico

«a

ls te

i oa a oe ee Reo A coos eg Lo x | =

No. 25. Zrachemys taylori (Legler, 1960). The Cuatro Ciénegas Slider is an endemic Mexican species restricted in distribution to the

Cuatro Ciénegas Basin (Lemos-Espinal et al. 2015). This individual came from Cuatrociénegas in the municipality of Cuatrociénegas

de Carranza. Wilson et al. (2013a) determined its EVS as 19, placing it in the upper portion of the high vulnerability category. Its

conservation status is calculated as Endangered by IUCN, but this species is not listed by SEMARNAT. Photo by Michael S. Price.

state support the most significant herpetofaunas based

on the relative numbers of country endemics and high

vulnerability species. Three such areas were identified:

the Gran Sierra Plegada, the Sierras Transversales, and

the Sierras y Llanuras Coahuilenses. Fortunately, these

three regions support the greatest numbers of protected

areas among the 19 that are currently established, four,

four, and eight, respectively. The herpetofaunal content

of these protected areas, however, is very poorly known;

as a result, the major conservation goal with respect to

the herpetofauna of Coahuila is to carefully document

the species inhabiting the protected areas of the state in

order to test the predictions made here about their content

and to draw up adequate management plans for their

perpetual protection.

B. Thus, it will only be after the species inhabiting

the existing protected areas have been identified that

additional conservation goals can be addressed. These

goals include (1) determining what other protected areas

might need to be established to protect the remainder of

the herpetofauna not found within the existing areas, (2)

monitoring of the health of the populations of species

within the protected areas, and (3) assessing the well-

being of the ecosystems on which these species depend.

C. It is imperative that this work advance as rapidly

as possible, especially given that efforts to protect

Amphib. Reptile Conserv.

the Coahuilan herpetofauna lag behind those of the

other Mexican states examined thus far in the Mexican

Conservation Series.

“We have come a very long way through the barbaric

period in which we still live, and now I believe we have

learned enough to adopt a transcendent moral precept

concerning the rest of life. It is simple and easy to say:

Do no further harm to the biosphere.” —E.O. Wilson

(2016)

Acknowledgments.—We are very thankful to those

individuals who allowed us to use their outstanding

photographic images of many of the amphibians,

reptiles, ecosystems and environmental issues illustrated

in this paper, including: Michael S. Price; Marco Antonio

Bazan-Tellez; Uri Garcia- Vazquez; Daniel Garza Tobon;

Bernardo Marino (http://gransierraplegada.org); Gabriel

Viesca Ramos; José Flores Ventura; and Daniel Solorio

Estrada. We are indebted to Dr. José Juan Flores from

Especies, Sociedad y Habitat A. C., for constructing the

physiographic map.

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Gonzalez-Sanchez VH, Johnson JD, Garcia-Padilla E,

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Johnson JD, Mata-Silva V, Garcia-Padilla E, Wilson

LD. 2015a. The herpetofauna of Chiapas, Mexico:

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Hernandez R, Navarro-Velazquez B, Wilson LD,

Powell GL, Russell AP. 2017. Texas Horned Lizards

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JD, Wilson LD. 2019. The endemic herpetofauna of

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2014. Coalescent species delimitation in milksnakes

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Life. Liveright Publishing Corporation, New York,

New York, USA. 259 p.

Wilson LD, McCranie JR. 2004. The conservation

status of the herpetofauna of Honduras. Amphibian &

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conservation reassessment of the reptiles of Mexico

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Conservation 7(1): 97-127 (e69).

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Padilla E, Wilson LD. 2016. The herpetofauna of

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herpetofauna of Puebla, Mexico: composition,

distribution, and conservation status. Mesoamerican

Herpetology 4: 790-884.

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Amphib. Reptile Conserv.

Lazcano et al.

David Lazcano is a herpetologist who earned a bachelor’s degree in chemical science in

1980, and a bachelor’s degree in biology in 1982. In 1999, David earned a master’s degree

in wildlife management, and a doctoral degree in biological sciences with a specialty

in wildlife management (2005), all from the Facultad de Ciencias Bioldgicas of the

Universidad Autonoma de Nuevo Leén (UANL). Currently, he is a full-time professor

j at the same institution, where he teaches courses in animal behavior, biogeography,

biology of chordates, and wildlife management. David is also the head of Laboratorio

de Herpetologia and Coordinacion de Intercambio Académico de la Facultad de Ciencias

Biolégicas at UANL. Since 1979, he has been teaching and providing assistance in

both undergraduate and graduate programs. David’s research interests include the

, herpetofaunal diversity of northeastern Mexico, as well as ecology, herpetology, biology

of the chordates, biogeography, animal behavior, and population maintenance techniques

of montane herpetofauna.

Manuel Nevarez-de los Reyes is a biologist who graduated from the Universidad

Autonoma de Nuevo Leon (UANL), Facultad de Ciencias Biologicas in San Nicolas

de los Garza, México. Manuel’s initial interest was in the study of amphibians and

reptiles, but his professional life led him to investigate other areas, such as environmental

impacts and the study of cacti. From 1997 to 2007, he served as head of Environmental

Protection in the Residencia Regional de Construccion Noreste of the Federal Electricity

Commission. Manuel has been involved with numerous workshops and conferences, and

has authored both popular science and peer-reviewed articles on herpetology and cacti.

Among his accomplishments, he discovered and co-authored the original description of

a new genus and species of cactus, Digitostigma caput-medusae. The following year he

created “Proyecto Digitostigma,” a nursery dedicated to the commercial propagation of

various cacti, which contributes to their knowledge and conservation. Manuel recently

obtained his Ph.D. in Wildlife Management and Sustainable Development at the UANL,

with a thesis entitled “Ecological distribution of the herpetofauna of the Sierra de Gomas

in northern Nuevo Leon,” under a grant from the National Council of Science and

Technology. He is now part of the herpetological group that has documented the many

herpetological activities in northeastern México (Coahuila, Nuevo Léon, and Estado de

México).

Eli Garcia-Padilla is a herpetologist primarily focused on the ecology and natural history

of the Mexican herpetofauna. His research efforts have centered on the Mexican states

of Baja California, Tamaulipas, Chiapas, and Oaxaca. His first experience in the field

was studying the ecology of the insular endemic populations of the rattlesnakes Crotalus

catalinensis, C. muertensis (C. pyrrhus), and C. tortugensis (C. atrox) in the Gulf of

California. Eli’s Bachelor’s thesis was on the ecology of C. muertensis (C. pyrrhus) on

Isla El Muerto, Baja California, Mexico. To date, he has authored or co-authored over

100 contributions to science. Eli is currently a formal Curator of Amphibians and Reptiles

from Mexico in the electronic platform “Naturalista” of the Comision Nacional para el

Uso y Conocimiento de la Biodiversidad (CONABIO-inaturalist; http://www.naturalista.

mx). One of his main passions is environmental education, and for several years he has

worked on various projects that include the use of photography and audiovisual media as a

powerful tool for reaching large audiences and promoting the knowledge, protection, and

conservation of Mexican biodiversity. Eli’s interests include wildlife and conservation

photography, and his art has been published in several scientific, artistic, and educational

books, magazines, and websites. Presently, he is collaborating on an evaluation of the

jaguar (Panthera onca) as an umbrella species for the conservation of the herpetofauna of

Nuclear Central America.

Jerry D. Johnson is Professor of Biological Sciences at The University of Texas at El

Paso (UTEP), and has been investigating the systematics, ecology, and conservation of

the herpetofauna of Middle America since 1970, especially that of southern Mexico.

Jerry is also the Director of UTEP’s 40,000-acre Indio Mountains Research Station in

the Chihuahuan Desert of Trans-Pecos, Texas. He has authored or co-authored over 120

peer-reviewed papers, and was co-editor or contributor to several major Mesoamerican

herpetology books: Conservation of Mesoamerican Amphibians and_ Reptiles,

Mesoamerican Herpetology: Systematics, Zoogeography, and Conservation, and Middle

American Herpetology: A Bibliographic Checklist. One species, Tantilla johnsoni, was

named in his honor. Presently, Jerry is an Associate Editor and Co-chair of the Taxonomic

Board of the Mesoamerican Herpetology website.

93 October 2019 | Volume 13 | Number 2 | e189

Amphib. Reptile Conserv.

The herpetofauna of Coahuila, Mexico

Vicente Mata-Silva is a herpetologist originally from Rio Grande, Oaxaca, Mexico.

His interests include ecology, conservation, natural history, and biogeography of the

herpetofaunas of Mexico, Central America, and the southwestern United States. Vicente

received his B.S. degree from the Universidad Nacional Autonoma de México (UNAM),

and his M.S. and Ph.D. degrees from the University of Texas at El Paso (UTEP). Vicente

is an Assistant Professor of Biological Sciences at UTEP in the Ecology and Evolutionary

Biology Program, and Assistant Director of UTEP’s 40,000 acre Indio Mountains Research

Station, located in the Chihuahuan Desert of Trans-Pecos, Texas. To date, Vicente has

authored or co-authored over 100 peer-reviewed scientific publications. He also was the

Distribution Notes Section Editor for the journal Mesoamerican Herpetology.

Dominic L. DeSantis is currently a Ph.D. candidate and National Science Foundation

Graduate Research Fellow at the University of Texas at El Paso. He received his Bachelor’s

degree at Texas State University, where he also completed multiple research projects on the

antipredator behavior of the critically endangered Barton Springs Salamander (Eurycea

sosorum). Dominic’s ongoing dissertation research integrates multiple field monitoring

technologies to study snake movement and behavioral ecology. Dominic accompanied

Vicente Mata-Silva, Eli Garcia-Padilla, and Larry David Wilson on survey and collecting

trips to Oaxaca in 2015, 2016, and 2017, and he is a co-author on numerous natural history

publications produced from those visits.

Larry David Wilson is a herpetologist with lengthy experience in Mesoamerica. He was

born in Taylorville, Illinois, USA, and received his university education at the University

of Illinois at Champaign-Urbana (B.S. degree) and at Louisiana State University in

Baton Rouge (M.S. and Ph.D. degrees). He has authored or co-authored over 410 peer-

reviewed papers and books on herpetology. Larry was the senior editor or author of several

books, including Conservation of Mesoamerican Amphibians and Reptiles, The Snakes of

Honduras, Middle American Herpetology, The Amphibians of Honduras, Amphibians &

Reptiles of the Bay Islands and Cayos Cochinos, Honduras, The Amphibians and Reptiles

of the Honduran Mosquitia, and Guide to the Amphibians & Reptiles of Cusuco National

Park, Honduras. To date, he has authored or co-authored the descriptions of 72 currently

recognized herpetofaunal species, and seven species have been named in his honor,

including the anuran Craugastor lauraster, the lizard Norops wilsoni, and the snakes

Oxybelis wilsoni, Myriopholis wilsoni, and Cerrophidion wilsoni. Larry previously served

an Associate Editor and is presently Co-chair of the Taxonomic Board for the journal

Mesoamerican Herpetology.

94 October 2019 | Volume 13 | Number 2 | e189

Official journal website:

amphibian-reptile-conservation.org

Amphibian & Reptile Conservation

13(2) [General Section]: 95-101 (e190).

Unpublished population data of Dendrobates azureus

Hoogmoed 1969 obtained in 1968 and 1970, and its historical

and current taxonomic status

Marinus S. Hoogmoed

Museu Paraense Emilio Goeldi, Caixa Postal 399, 66017—-970 Belém, Pard, BRAZIL

Abstract.—During a herpetological inventory in the Sipaliwini area in southern Suriname in 1968, and again

during a second expedition to the area in 1970, anecdotal population data on Dendrobates azureus Hoogmoed,

1969 were obtained. As of now, some 50 years later, these data have not been published, yet they may be useful

for the evaluation of the status of this taxon at the present time and the evolution of its populations over the

period since 1968. Visits to the Sipaliwini savanna to observe or collect this taxon over the past 50 years have

been few and far between. An overview of the population data available in the publications about these visits

is provided.

Keywords. Dendrobates tinctorius, isolated populations, Sipaliwini savanna, Suriname, conservation, threats.

Citation: Hoogmoed MS. 2019. Unpublished population data of Dendrobates azureus Hoogmoed 1969 obtained in 1968 and 1970, and its historical

and current taxonomic status. Amphibian & Reptile Conservation 13(2) [General Section]: 95-101 (e190).

4.0 International (CC BY 4.0): https://creativecommons.org/licenses/by/4.0/], which permits unrestricted use, distribution, and reproduction in any

medium, provided the original author and source are credited. The official and authorized publication credit sources, which will be duly enforced, are

as follows: official journal title Amphibian & Reptile Conservation; official journal website: amphibian-reptile-conservation.org.

Received: 14 March 2018; Accepted: 10 September 2019; Published: 9 October 2019.

Introduction it seems timely to publish the basic population data of

this taxon, even though they were collected anecdotally.

During a herpetological inventory of the Sipaliwini area Polder (1974), based on his observations of the taxon

in 1968 Hoogmoed (1969a,b, 1971a,b, 1972) discovered __in captivity, expressed some doubt on the specific status

a blue Dendrobatid frog in forest islands in the Sipaliwinit of D. azureus, but this had no direct consequences as

savanna, near the Vier Gebroeders Mountain close to the — Silverstone (1975) considered it a species. Wollenberg

Suriname-Brazilian border, that he named Dendrobates et al. (2006), followed by Wollenberg (2007), on the

azureus. In the description (Hoogmoed 1969b),a number basis of morphological data and genetic analysis,

of ecological and behavioral data were provided, but synonymized D. azureus with D. tinctorius (Cuvier,

data that give an impression about the abundance of the 1797). This assessment was supported by Noonan

population were not presented, because they were not and Gaucher (2006), who found that specimens of D.

collected in a systematic way, but rather anecdotally. azureus (from the type locality or near to it, see below,

The author paid a second visit to the Sipaliwini area and Noonan, pers. comm.) shared the same haplotype with

the habitat of D. azureus in 1970 (Hoogmoed 1971a,b, _ two “nearby” (from the continuous rainforest, see below,

1972) and more data were obtained on the abundance = Noonan, pers. comm.) populations of D. tinctorius and

of the frog in the type locality (forest island on West — concluded that this signified the two taxa were identical.

slope Vier Gebroeders Mountain) and some other forest Grant et al. (2006) still treated D. azureus as a species

islands nearby, and about forest island occupation by this —_ but on the basis of molecular results were inclined to

taxon. These population data are available inthe author’s _— follow the synonymy suggested by Wollenberg et al.

field books (archived in the former Rijksmuseum van = (2006). Gaucher and McCulloch (2010) and Frost (2018)

Natuurlijke Historie, now Naturalis Biodiversity Center accepted the synonymization and considered D. azureus

in Leiden [RMNH], in the Netherlands) and in his private — a synonym of D. tinctorius. Ouboter and Jairam (2012)

diary notes, and they can provide basic data about the considered D. azureus a subspecies of D. tinctorius,

population status of the taxon at the time of its discovery. —_ without providing arguments, but this was not accepted

This taxon is considered vulnerable [VU: D2] (Stuart et by Frost (2018), who treated D. tinctorius as monotypic.

al. 2006), and as a member of the genus Dendrobates it is However, Avila-Pires et al. (2010) and Hoogmoed

on CITES Appendix II, so in Suriname it officially cannot —_ (2013) did not accept the synonymization by Wollenberg

be traded (Hoogmoed 2013). However, since it still has et al. (2006) and pointed out that this publication suffered

been subject to illegal capture and export of specimens, several shortcomings. Here I add to those critiques the

Correspondence. marinus@museu-goeldi.br

Amphib. Reptile Conserv. 95 October 2019 | Volume 13 | Number 2 | e190

Dendrobates azureus in Suriname 1968-1970

use of tissue samples of captive bred material of dubious

origin (although stated to be from the type locality) and

the fact that among dendrobatid breeders D. azureus has

been confused with, and interbred with, a blue morph of D.

tinctorius that occurs in southern Guyana, northwestern

Para in Brazil (south to Porto Trombetas) and possibly

in extreme southwestern Suriname (in a contested area

between Suriname and Guyana). See Avila-Pires et al

(2010: Fig. 23) and Lotters et al. (2007: Fig. 707) for

color pictures of this blue D. tinctorius morph. Hoogmoed

(1969, 2013: Fig. 4), Eisenberg (2004), and Lotters et al.

(2007: Fig. 708 [as D. tinctorius|) provide pictures of D.

azureus. Noonan and Gaucher (2006) mention specimens

of D. tinctorius and D. azureus from slightly different

localities (Table 1) and their samples of D. azureus are

from the area of Vier Gebroeders Mountain (see Noonan,

pers. comm., below as well). In Wollenberg et al. (2008) it

seems that the blue morph of D. tinctorius has incorrectly

been considered as D. azureus (see Fig. 4 Haplotype 2,

the second figure from above and the lower figure; no

D. azureus can be found in this figure). Unfortunately,

only the fancy names of hobbyists have been used for

the “Sipaliwini” material and no vouchers have been

indicated.

In order to remove any doubt regarding which

population I am discussing, and avoid upsetting the

present accepted nomenclature, below I use the name

Dendrobates “azureus” in the sense of the population of

D. tinctorius described in 1969 as D. azureus and only

known from isolated forest islands in the Sipaliwini

savanna in southern Suriname.

History of (the herpetological) exploration of

the Sipaliwini Savanna and inventories of D.

“azureus” populations

1935-1938: Border expedition (van Lynden 1939)

to establish the border between Suriname and Brazil

(= watershed). Although Van Lynden stayed on the

Sipaliwini savanna for an extended period (4 October

1935 to 10 March 1936), and made general observations

about animals (mostly mammals and birds), he did not

mention “blue frogs,” so we may conclude he did not

observe them, or that he did not deem them worthy of

mentioning. His camp III (on the western base of Vier

Gebroeders Mountain, from where he wrote his diary

on 12 October 1935) actually was close to the later

type locality of D. “azureus,” but it was probably in the

savanna itself, not in the forest island. The map published

by van Lynden (1939) unfortunately does not show the

location of the forest islands.

1961: During Operation Grasshopper, the Sipaliwini

airstrip was constructed on a small savanna about 3.5

km west of the western border of the Sipaliwini savanna.

Apparently, the forest islands in the savanna were not

visited.

1968: Hoogmoed (1969a,b) discovered populations of

D. “azureus” in four forest islands in the middle of the

Sipaliwini savanna near the Vier Gebroeders Mountain

Amphib. Reptile Conserv.

Fig. 1. Dendrobates “azureus” (= tinctorius).

when making a herpetological inventory of the area in

1968 (22 August to 7 October), concentrating on the

area between Sipaliwini airstrip and Vier Gebroeders

Mountain. He collected a total of 37 specimens and

two tadpoles of D. “azureus” (Table 1), which were all

preserved and form the type material of the description of

D. azureus Hoogmoed, 1969. The location and shape of

the forest islands as shown in Hoogmoed (1969b: Fig. 3)

were taken from a topographical map (Centraal Bureau

Luchtkartering, Paramaribo, Suriname) of the area based

on aerial photographs.

1968-1969: After Hoogmoed’s departure from the area,

the Sipaliwini expedition (Hoogmoed 1969a) continued

working in the Sipaliwini savanna and the participants

(botanists and a geologist) moved N of the Vier

Gebroeders area to the Morro Grande Mountain area.

Hoogmoed (1969b: Fig. 3) marked four northern forest

islands as possible localities for D. “azureus” because

the botanist J.P. Schulz reported having seen “blue frogs”

there. Considering later observations (see Gagliardo 2004

a,b; Fouquet et al. 2015), these might well have been D.

tinctorius with a yellow semicircular mark on the snout.

1970: Between 13 January and 13 February Hoogmoed

again visited the Sipaliwini savanna, this time

concentrating on the part south of the Vier Gebroeders

Mountain and the Brazilian border. During this trip one

specimen of D. “azureus” was collected and preserved,

and 10 additional specimens were collected at the type

locality and transported alive to the Netherlands, where

they were bred in captivity by Polder (1973a-—c, 1974).

1981: According to Wevers (2007), a “group of

Dutchmen” brought live D. “azureus” from the Sipaliwini

savanna (most likely an illegal operation), making no

mention of the names of participants, numbers of frogs

brought back, or from which forest island(s).

1988: An illegal import of D. “azureus” (apparently

three specimens) was confiscated in the Netherlands and

transferred to Blijdorp Zoo, Rotterdam (Wevers 2007).

1996: Cover (1996, 1997) reported on an expedition

in June 1996 (sponsored by the National Aquarium in

Baltimore, USA) to attempt a population survey of D.

October 2019 | Volume 13 | Number 2 | e190

Hoogmoed

Table 1. Detailed data for specimens of D. “azureus” collected/observed by M.S. Hoogmoed in 1968 and 1970. Number of

specimens accounts for both collected and observed specimens, numbers between brackets in the first column refer to the numbers

of forest islands as used in the text. The asterisk (*) indicates that 10 specimens were observed in about 10 minutes.

Forest

island Number of specimens | Time spent in field

(see text)

Person-minutes Number person-

spent in field minutes per specimen

1968

5 (+ 2 spol

au GUN BE) — ——

23 Sep 10 ——— coll, RMN a 00-11:00 h

=a Mean or total

1970

A CT ——

Soa eae

Total 1968 + 1970

82 (38 preserved, 10

live for ex situ breeding

colony)

“azureus.” Three staff members of the NAIB and three _— (and finally exported) in a forest island northeast of the

field workers of Conservation International Suriname Sipaliwini airstrip. One day before departure from the

participated. Fifty-four adults and two juveniles were = savanna permission was obtained to collect and export

observed during a limited number of days. They surveyed 20 specimens of D. “azureus,” so they had to be collected

six forest islands and found specimens in three of them at arush and were meant to establish an ex-situ breeding

“two on the slopes of Vier Gebroeders and one ina valley —_ population (Eiben 2005). No mention was made of which

floor forest just north of the mountains” (Cover 1997; forest island(s) these specimens were collected from or

Eiben 2005). Most likely Cover referred to forest island |= how much time it took to collect them.

nos. 1 and 4 on the slopes of Vier Gebroeders Mountain

and to forest island no. 2 north of that mountain (see 2003: B.P. Noonan (pers. comm.; Eiben 2005) visited

below). No material was collected, and the position of — the area of the Vier Gebroeders Mountain from 23—26

the other three forest islands was not mentioned. May 2003. He flew in using Mamiya airstrip (= “Myers’

airstrip” in Hoogmoed (1969) and in the present text,

1997: Gagliardo (2004a,b) reported on a new expedition Wapaisana Anotato on Google Earth) on the border

(14 August-19 September 1997) by Cover and three of Suriname and Brazil, SE of the Vier Gebroeders

other zoo curators in order to collect specimens to Mountain, and he left via Sipaliwini airstrip, W of the

establish a breeding population in the USA. “Nearly 60 — savanna. He was not allowed to collect specimens of D.

specimens” and an unknown number of tadpoles were “azureus,” but was allowed to make toe clips from 10

observed in two forest islands that were not the type — specimens for molecular studies (Noonan and Gaucher

locality. Furthermore, three pairs of D. tinctorius (witha 2006). He found three specimens on 24 May, four

yellow semi-circular mark on the snout) were collected = specimens on 25 May, and another three on 26 May. No

Amphib. Reptile Conserv. 97 October 2019 | Volume 13 | Number 2 | e190

Dendrobates azureus in Suriname 1968-1970

data on time spent finding specimens are available, but

Noonan writes: “....my experience was that neither of

these populations was terribly dense. While I did not

keep detailed notes on abundance, I am comfortable

saying that I did not observe more than one individual per

hour of searching (on average).” Noonan also collected

tissue from three Dendrobates specimens found in the

continuous rainforest NE of the airstrip Sipaliwini, about

100 m from the savanna edge, that were identified as D.

tinctorius (Noonan and Gaucher 2006).

2007: According to Wevers (2007) several frog fanciers

visited the Sipaliwini savanna and “observed respectively

9, 15 [probably Wevers himself] and 20 specimens

(mostly the same specimens).” Wevers (2007) visited the

Sipaliwini savanna for five days in February and during

those days observed 15 specimens and four larvae. He

at least visited the type locality on the western slope of

the Vier Gebroeders Mountain and the forest island on

the northeastern slope from where he reported juvenile

specimens. He reported that his guide who lived on

Mamiya airstrip (= “Myers’ airstrip”) on the frontier of

Suriname and Brazil, never had seen more than 25 D.

“azureus”’ in one day. Based on his own observations

(five days and a limited number of forest islands visited)

and information from his Indian guides, he estimated

the size of the total wild population to be between 1,000

and 1,500 specimens, but this does not seem to be a very

reliable figure.

2014: Fouquet et al. (2015) visited the Sipaliwini area

between 15 and 28 April, but did not visit the forest

islands where D. “azureus” occurs. They reported D.

tinctorius (with a yellow semi-circular mark on the

snout) from a mountain 10 km N of Sipaliwini airstrip in

the area of continuous rainforest.

By no means is this overview intended to be an

exhaustive listing of all visits to the D. “azureus” habitat

or nearby areas. It is known that Suriname scientists

with a license to study the frogs and personnel of the

Forestry Service flew into Sipaliwini airstrip, but were

not allowed to travel from there to the Vier Gebroeders

Mountain and they were confined to the airstrip. Some

scientist may have paid unregistered (and unpublished)

visits to the area. Illegal collectors (animal dealers) and

terrarium keepers apparently have visited the area at

least several times, but because of the nature of these

trips, they have not been documented publicly. It also is

possible that native and Brazilian Indian collectors have

provided animal dealers with specimens that may have

left Suriname directly or via Brazil. No numerical data

are available, but Suriname animal dealers exporting

reptiles and amphibians to the USA and Europe have

long-standing commercial contacts (since the early

1970’s) with the Indians of the villages of Alalapadu and

Kwamalasemutu.

Material and Methods

During the 1968 herpetological inventory of the Sipaliwini

savanna in southern Suriname (Hoogmoed 1969a,b), five

forest islands near the Vier Gebroeders Mountain were

Amphib. Reptile Conserv.

searched. In 1970, Hoogmoed surveyed five forest islands

in the southern part of the savanna. During the fieldwork

in the area around Vier Gebroeders Mountain no formal

population surveys were made, but notes were kept about

how many specimens were observed/collected during

the time spent along transects in the forest islands. Frogs

were observed/collected while traversing forest islands

following creek beds, either downhill or ascending the

creek, generally searching an area of five m at each side

of the stream. The time period during which frogs were

collected was noted, and based on this the abundance

was expressed in specimens per person-minutes. In 1968

observations/collections were made by one person, and

in 1970 by two people. All specimens were either simply

observed, or collected by hand. Specimens collected were

killed with MS222, fixed and preserved in 70% ethanol

(thus, no formaldehyde was used and the type specimens

could still be used for DNA analysis). Live specimens were

transported in plastic bags with leaf litter, and termites

were provided as food.

Data on specimens collected in 1968 were provided

by Hoogmoed (1969b) in general terms. The coordinates

of forest islands where D. “azureus’” was found were

calculated in 1968 on the basis of a topographic map of

the area, but they now can be provided more precisely,

based on localization with Google maps. Only slight

differences can be noticed.

In 1968, the area of the Sipaliwini savanna where

D. “azureus” was obtained was visited between 11

September and 1 October, a total of 21 days. During

this period, several forest islands on and near the Vier

Gebroeders Mountain (as well as the intervening savanna

area) were searched for herpetofauna. Coordinates for

the center of the forest islands are given as in Hoogmoed

(1969b) and corrected according to Google Earth 2018,

datum W84. Forest islands inhabited by D. “azureus” are

indicated with asterisks (*).

1. *Forest island W flank Vier Gebroeders Mountain

(Base Bivouac), type locality of D. azureus,

2°N, 55°58’W (corrected to 2°00’21.24°N,

55°58°10.85”W)

2. *Forest island (J-shaped [the eastern narrow

extension is not rainforest but gallery forest of

Manritia palms]) 1.5 km NE of Vier Gebroeders

Mountain, 2°01’N, 55°57.30’W (corrected to

2°00’59.30”N, 55°57’ 26.03” W)

3. *Forest-island 2 km (note this distance differs

from that in the description of D. azureus) N of

Vier Gebroeders Mountain, long and narrow,

directed W—-E, 2°01’N, 55°58’W (corrected to

2°01°25.78"N, 55°57’°34.22”W)

4. *Forest island on NE slope Vier Gebroeders

Mountain, 2°N, 55°57’30"W (corrected to:

2°00’24.92”N, 55°57’ 22.03” W)

5. Forest island (small) on N slope Vier Gebroeders

Mountain (Google Earth 2°007°52.49"N,

55°58’°04.71”W)

In 1970, the Sipaliwini savanna was visited again (13

January—13 February), this time mostly in a part further

south from the area visited in 1968, with a stay of only

October 2019 | Volume 13 | Number 2 | e190

Hoogmoed

two days in forest island no. 1 (see above). During this

period the following forest islands, that turned out not to

be inhabited by D. “azureus,” were searched (coordinates

based on Google Earth 2018):

6. Small forest island on northernmost part of Lange

Dijk, 2°00’09.36”N, 55°55’°58.78”W, Suriname,

27 January 1970

Small forest island on ridge of Lange Dijk,

1°59°28.01”"N, 55°55715.81”W, Brazil, 27

January 1970

Elongate forest island on SW slope Lange Dijk,

1°59°18.74"N, 55°55’20.73”W, Brazil, 27

January 1970

Westernmost small forest island of two, E

of Myers’ farm, about 8 km WSW of Vier

Gebroeders Mountain W flank, 1°59’°09.10’N,

56°02’35.11”W, Suriname, 4 February 1970

Easternmost (630 m E of and twice as large as

no. 9) forest island, E. of Myers’ farm, about 8

km WSW of Vier Gebroeders Mountain W flank,

1°59°11.78"N, 56°02713.63”W, Suriname, 4

February 1970.

10.

Results

Forest Islands

The shapes and sizes of eight forest islands in 1968 were

based on a topographical map and aerial photographs

that formed the basis for Figure 3 in Hoogmoed (1969b).

These were all compared with Google Earth images of 31

December 1969 and 17 November 2004 (the most recent

freely available large-scale images on the Internet), and

all the forest islands still exist and no notable changes

in shape or size were observed. Cover (1997) noted that

concern had been expressed that the anthropogenic fires

which ravage the savanna yearly might damage the forest

islands. In 1970, Hoogmoed observed that savanna fire

had destroyed the narrow band of forest between the

1968 Vier Gebroeders Bivouac (type locality) and the

savanna on the SW edge of the campground, but also

that the forest island interior, probably because it 1s

rather moist, had not suffered any damage. D. “azureus”

was still regularly present in the former, open camp

ground. Gagliardi (2004a,b) stated that the fires did enter

the forest islands, but he did not mention the extent of

damage. Wevers (2007) wrote that fire had gnawed at the

edges of the forest islands and expressed fear that in an El

Nifio year fire might reach the interior of the forest islands

and thus threaten their integrity. Cover (1997), however,

reported that the fires apparently did not damage the

forest islands, but thought that they might be the reason

that the forest islands did not expand into the savanna.

These last observations are confirmed by Hoogmoed’s

1970 observations (Hoogmoed 1972) and by the Google

Earth images of 2004. The Map for Environment (2018)

shows that there has been only limited tree loss in the

Sipaliwini savanna between 2000 and 2014, although

tree loss near Sipaliwini airstrip has been significant. The

forest islands themselves do not show any noticeable

changes.

Amphib. Reptile Conserv.

The most recent Google Earth images (2004) show

that in the Brazilian part of the savanna (Paru savanna)

south and east of the Sipaliwini savanna there are six

large forest islands that would be worth investigating

for the presence of D. “azureus.” However, as this area

is a Brazilian Indian Territory, conducting biological

research there is very difficult, because of the need for

special permits and its remoteness. Just south of the Paru

savanna in Brazil is another, isolated, more or less oval

savanna with a large, elongate forest island in the middle

(160 km SSW of Sipaliwini airstrip). This forest island

was inventoried by Avila-Pires et al. (2010: ESEC Grao

Para Centro) and they did not find any Dendrobates

species there.

Population Data

11 September—1 October 1968. For this period of 21

days spent in Vier Gebroeders Bivouac, general herpe-

tological collecting was conducted in the savanna and

forest islands on and near Vier Gebroeders Mountain.

Only some parts of the days were spent in forest islands

searching for D. “azureus.”

Apart from the four forest islands where specimens

were observed and collected, one small forest island on

the N slope of Vier Gebroeders Mountain was searched

for D. “azureus,” but no specimens were found. The four

northernmost forest islands in Hoogmoed’s (1969) map

were not visited during this time. Data on time spent ob-

serving/collecting D. “azureus” and population density

are summarized in Table 1.

Between 1968 and 1970. The population density of D.

“azureus’ in forest island no. 1 on the W slope of the Vier

Gebroeders Mountain seems to have diminished consid-

erably (remembering that 23 specimens and two tadpoles

were removed in 1968, which might have had a negative

influence on the population), viz. one specimen per 4.7

person-minutes in 1968 (one observer only), versus one

specimen per 19.2 person-minutes in 1970 (two observ-

ers).

In 1970 an additional 11 specimens were removed

from this same forest island, one for the RMNH collec-

tion, and ten live specimens to establish an ex-situ breed-

ing colony in the Netherlands.

It should be mentioned that specimens were not even-

ly distributed throughout the forest islands. They might

be absent in certain stretches and be numerous in other

parts (generally near creeks and/or in areas with large

boulders).

No comparative data for the other forest-islands are

available for the period 1968-1970. Cover (1996, 1997)

does not provide data in a comparable way, but he ap-

parently collected data in three of the forest islands men-

tioned by Hoogmoed (1969), but unfortunately the lo-

cations of these have not been published. Noonan (pers.

comm. 2017) reported that he did not see more than one

specimen per hour. Already in 1968, the population den-

October 2019 | Volume 13 | Number 2 | e190

Dendrobates azureus in Suriname 1968-1970

sity in the other forest islands seemed to be less (one

specimen per 18—30 person-minutes) than in the largest

forest island (no. 1) on the W slope of Vier Gebroeders

Mountain (one specimen per 4.7 person-minutes). This

could be related to the size of the forest islands, but this

is Just an impression that is not based on firm facts. Also,

we have to take into account that more time was spent in

forest island no 1, because it was the location of the camp

(both in 1968 and 1970).

Collections in 1968 were made during the second part

of September, during the dry season, and those in 1970

were made in early February, during the beginning of the

wet season—when rainfall is about twice that in the dry

season, and about half that of May and June, the wettest

months (see Hoogmoed 1969).

Conclusions

At the time of the discovery of D. “azureus” in 1968

it was clear that not all forest islands inhabited by this

taxon had populations of the same density. Since 1968,

although several expeditions have visited the distribution

area of D. “azureus” in the Sipaliwini savanna, no

data on population densities have been published that

could be directly compared with those presented here.

However, the anecdotal data available (see the History

of ... inventories of D. “azureus” populations section

above) give the strong impression that the numbers of

D. “azureus” in its restricted habitat have considerably

diminished since 1968. This impression should be

confirmed by systematic population studies that might

serve in situ and ex situ management programs for this

unique population of brilliant blue poison frogs. At the

moment we do not even have an idea about the size of

the population in the wild, but it might run only into the

hundreds. Eiben (2005) and Stuart et al. (2008) described

the successful ex situ breeding program in the National

Aquarium in Baltimore (Maryland, USA) based on 20

specimens collected in 1997 (see above) and some

additional exchanged specimens. This program should

be continued and fortified with the help of the Suriname

authorities and several nature conservation interest

groups, such as WWE, The Nature Conservancy, and

Conservation International, that are already active in

Suriname.

Although the habitat of this taxon 1s completely within

a Suriname Nature Reserve, the area is easily accessible

from Brazil and the border is not patrolled. Illegal visits

by collectors cannot be ignored, and should be taken into

account when making an in situ management plan. Stuart

et al. (2008: 228) optimistically assumed that interest in

wild collected specimens would diminish with successful

breeding in captivity, but this is a naive assumption (e.g.,

IUCN 2015).

Acknowledgements.—Trips to the Sipaliwini savanna

were made in close cooperation with the Suriname

Forest Service (Dienst’s Landsbosbeheer, F. Bubberman,

J.P. Schulz), which supplied laborers and equipment.

Fieldwork in 1968 was funded by a grant (WR 956—

2) from WOTRO (Netherlands Foundation for the

Amphib. Reptile Conserv.

Advancement of Tropical Research). Fieldwork in 1970

was supported by grants from the Foundation Jan Joost

ter Pelkwijk and the legacy of Miss A.M. Buitendijk,

both administrated by the Rijksmuseum van Natuurlijke

Historie, Leiden, Netherlands.

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het Palmblad 10: 14—23.

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Marinus Steven Hoogmoed was curator of Herpetology at the Dutch

National Museum of Natural History (RMNH) in Leiden from 1966

to 2004, and Head of its Department of Vertebrates from 1991 to 2001.

» Marinus obtained his doctorate degree in Mathematics and Natural

~ Sciences at Leiden University, Netherlands, in 1973 based on a monograph

2 of the lizards and amphisbaenians of Suriname. He worked mainly on

systematics, taxonomy, and biogeography of Amazonian and Guianan

® herpetofauna; and he has done fieldwork in all Amazonian countries, except

Guyana. Marinus spent a total of three years in the field in Suriname. After

his retirement, Marinus continued his research in the Amazon area as a

volunteer at the Museu Paraense Emilio Goeldi (MPEG) in Belém, Para,

Brazil, where he is still active. Between 1975 and 2004 he was involved in

CITES as a representative of the Netherlands, and between 2000 and 2002

Marinus was chair of the Animals Committee of CITES.

October 2019 | Volume 13 | Number 2 | e190

Official journal website:

amphibian-reptile-conservation.org

Amphibian & Reptile Conservation

13(2) [General Section]: 102-114 (e191).

Vulnerability of Northern Pine Snakes (Pituophis

melanoleucus Daudin, 1803) during fall den ingress

in New Jersey, USA

Joanna Burger

Division of Life Sciences, Rutgers University, 604 Allison Road, Piscataway, New Jersey 08854 USA

Abstract.—The management of threatened and endangered species often falls to various state agencies which

may have different and conflicting goals. The Pine Barrens of New Jersey are managed for different objectives,

including fire management, tree cutting, recreational activities (hiking, hunting, off-road-vehicle use), wildlife

protection, and conservation. Managing competing claims requires ecological information on critical issues and

vulnerabilities for determining the impacts of each claim. The Northern Pine Snake (Pituophis melanoleucus),

an iconic Pine Barrens species that is threatened in New Jersey, is normally dispersed during spring and

summer, but the snakes converge in the fall to communal hibernacula, where they spend the winter and leave

in the spring. Here, the activity of Northern Pine Snakes near hibernacula in the fall is described to examine

their vulnerability to various competing claims, such as fire or off-road vehicle use. Two hypotheses are tested:

(i) that snakes enter the hibernaculum once (and stay), and (ii) that the total period of ingress for all Northern

Pine Snakes is limited to just a few weeks in the fall. Activity of PIT-tagged snakes at hibernacula entrances

was monitored with a passive, continuously-recording AVID TracKer and temperatures were monitored with a

continuously recording thermometer placed at the soil surface. The behavior of marked snakes (18 in 2017, 25

in 2018), indicated that the period of activity around the hibernaculum entrance was: 1) longer than expected

(i.e., over two months), 2) involved multiple ingress and egress of individual snakes, and 3) sometimes involved

movement between two or among multiple nearby hibernacula. Northern Pine Snakes generally did not move in

or out of hibernacula when temperatures were below 9° C. Daytime high and nighttime low temperatures greatly

influenced movement. Although the daily high and low temperatures when snakes moved were correlated (r=

0.54 in 2017; 0.51 in 2018, P< 0.0001), the daily high and low temperatures were more highly correlated (r= 0.71

in 2017; 0.79 in 2018), indicating factors other than temperature influence snake activities. Most snakes entered

and exited between 1000 and 1800 h, although some moved as late as 0030 h. These data can inform science-

based decisions about when to allow tree cutting, fire management, and off-road vehicle races (e.g., increased

human activity). Most snakes are concentrated around hibernacula (but not necessarily near the entrances)

from early October until early December (or the end of December for two hatchlings). Therefore, a significant

proportion of snakes are vulnerable to disturbances that could impact their population viability. Vulnerabilities

are discussed in terms of competing claims and conservation.

Keywords. Behavior, competing claims, hibernation, reptiles, Serpentes, Squamata, wildlife management

Citation: Burger J. 2019. Vulnerability of Northern Pine Snakes (Pituophis melanoleucus Daudin, 1803) during fall den ingress in New Jersey, USA.

Amphibian & Reptile Conservation 13(2) [General Section]: 102-114 (e191).

ternational (CC BY 4.0): https://creativecommons.org/licenses/by/4.0/], which permits unrestricted use, distribution, and reproduction in any medium,

provided the original author and source are credited. The official and authorized publication credit sources, which will be duly enforced, are as follows:

official journal title Amphibian & Reptile Conservation; official journal website: amphibian-reptile-conservation.org.

Received: 14 January 2019; Accepted: 22 September 2019; Published: 6 November 2019

Introduction

Managers are often required to make environmental re-

source decisions with incomplete knowledge, with little

time, and often under conditions of competing claims for

resources and associated habitat. Human alteration of

natural lands is a key driver of global biodiversity loss

(Pimm and Raven 2000; Wilcove et al. 2000). Claims

for land can come from those who want to either use the

Correspondence. burger@dls.rutgers.edu

Amphib. Reptile Conserv.

resource itself (e.g., hunting, fishing, wildlife collect-

ing), use important components of the habitat (e.g., log-

ging), or improve the habitat for people (e.g., trail or road

building, fire suppression). Indeed, managers of differ-

ent resources (e.g., timber, wildlife) are often in conflict,

asserting competing claims for the same resource or hab-

itat. These need to be carefully considered and resolved

in a manner that reduces the risks to the habitat and

wildlife, while enhancing human benefits. For example,

November 2019 | Volume 13 | Number 2 | e191

Burger

managers of forests must balance cutting (or harvesting)

trees against re-forestation, the adverse effects of cutting

trees against the benefits of cutting them, and the relative

importance of the different benefits and costs (McLeod

and Gates 1998; Todd and Andrews 2008). Similarly, fire

suppression has some benefits (e.g., reduced potential for

fire damage to nearby communities and industries), as

well as costs if it does not occur (e.g., allowing dry debris

to build-up, creating the potential for a very hot canopy

fire when it does occur). Each option (e.g., logging, fire

suppression) has both benefits and costs to the plants and

animals living in the forests (Roth and Franklin 2018;

Steen et al. 2010). In addition, most animals face one or

more challenges to their survival, including predators,

competitors, poachers, and resource users, as well as

threats to their habitat from development, wildfire, or re-

source use. What may be good for one animal group may

not be good for another. For example, creating gaps in

forests may be good for deer management and for snakes

requiring open nesting areas, but is not good for interior-

nesting forest birds (Blouin-Demers and Weatherhead

2001; Gerald et al. 2006; MWPARC 2009). The complex

situations forest managers face can only be resolved by

examining the ecological and societal benefits and costs

of different options.

Setting priorities for conservation is a challenging

and necessary effort (Pimm et al. 2001). The biodiver-

sity crisis facing the Earth suggests that the conservation

needs of threatened and endangered species should be

considered first when managing habitats and ecosystems

(Wilson et al. 2009; Gaiarsa et al. 2015). Economic, soci-

etal, and political issues also play important roles in con-

servation decisions (Polasky 2008; Wilson et al. 2011).

However, it is equally important to understand the roles

of species vulnerability (UCN 2009), habitat loss (Wil-

son 1992: Pimm et al. 1995: Gibbons et al. 2000), habitat

fragmentation and patch size (Forman and Godron 1986;

Hilton-Taylor 2000; Kjoss and Litvaitis 2001; Sander-

son et al. 2002), restricted range or habitats (Segura et

al. 2007; Cardillo et al. 2008), human disturbances (Par-

ent and Weatherhead 2000), human infrastructures (e.g.,

roads, Andrews et al. 2008), and environmental stochas-

ticity (Tanentzap et al. 2012), among others (Gaiarsa et

al. 2015). Non-random distributions in time and space are

important aspects of vulnerability for animals, particular-

ly those that are slow moving or temperature-dependent

(Croak et al. 2013), including ectothermic vertebrates

(Kapfer et al. 2010). Understanding temporal and spatial

use of core areas 1s critical for conservation and manage-

ment (Semlitsch and Bodie 2003). All of these factors be-

come more important for understanding life histories and

conservation when they are considered within a frame-

work of human-related activities and impacts (e.g., fire

management, logging, development; Kapfer et al. 2010).

One of the iconic species of the New Jersey Pine Bar-

rens is the Northern Pine Snake (Pituophis melanoleucus

Daudin, 1803), a top-level predator that can reach 2 m in

Amphib. Reptile Conserv.

length. The Northern Pine Snake is listed as a threatened

species in New Jersey and as threatened or endangered

in other parts of its range in the southern United States

(Burger et al. 2017, 2018). New Jersey appears to have

the most stable population of this species (thus, a more

global responsibility for its conservation, Golden et al.

2009; Burger and Zappalorti 2016; Burger et al. 2017).

This paper examines the behavior of individually PIT-

tagged Northern Pine Snakes during their fall ingress

into hibernation sites (hibernacula or winter dens) in re-

lation to conservation of the species with respect to forest

management (e.g., logging, fire suppression) and other

human activities (e.g., poaching, off-road vehicle [ORV]

races, and traffic). There are competing claims for the

habitat (e.g., snake use and ORV use), for habitat man-

agement (e.g., fire management and deer management),

and for the snakes themselves (e.g., population stability

and poaching, Burger and Zappalorti 2016). The activi-

ties of snakes in the fall were monitored around several

hibernacula using new passive PIT-tag recording devices

located at the hibernaculum entrances. This technical ap-

plication is described with the intent of illustrating its use

for other such studies. The overall goal was to test the

null hypothesis (H,) that Northern Pine Snakes return to

their hibernacula and enter only once, remaining there

for the duration of the winter hibernation period. If H,

is rejected, this could indicate increased vulnerability in

time and space for a snake that is already threatened in

New Jersey.

Hibernation behavior in this species has been studied

previously in terms of hibernaculum site selection, use

and fidelity, structure of hibernacula, and the defensive

behavior of snakes disturbed during hibernation (Burger

et al. 1988, 2000; Burger and Zappalorti 2016, 2017).

Hibernaculum sites of other species of pine snakes (P.

ruthveni and P. melanoleucus lodingi) were studied in

Mississippi where the former used burrows of small

mammals, and the latter used decayed pine stumps and

roots for hibernation (Rudolph et al. 2007). Dispersal

rates around hibernation sites were also examined in

Gopher Snakes (Pituophis catenifer deserticola) in Brit-

ish Columbia (Williams et al. 2012). Some studies on

hibernation in other snake species include hibernation

site selection (Harvey and Weatherhead 2006), body

temperatures while hibernating (Costanzo 1986; Hein

and Guyer 2009), cold tolerance (Joy and Crews 1987),

factors affecting spring emergence (Todd et al. 2009),

spring emergence patterns (Hirth et al. 1969; Gregory

1974; Shine et al. 2006), and gene flow among snakes in

different hibernacula (Clark et al. 2008; Anderson 2010).

However, without individually marked snakes, and the

availability of the recent remote recording ability of the

Avid Power TracKer VIII, it was previously impossible

to determine the activity of individual snakes near hiber-

nation sites, the final entry temperatures for fall ingress,

or the final dates of entry into hibernacula, each of which

were incorporated into the present study.

November 2019 | Volume 13 | Number 2 | e191

Pituophis melanoleucus vulnerability during fall ingress

Temp

Reader

Den Entrance

PiTTag

Snake Cement Reader

Den Block (buried

Tunnel under soil)

Ss ya Data

Logger

Battery

12 Volts

Fig. 1. Schematic of deployment of the Tracker device at the entrance of a hibernaculum.

Methods

Northern Pine Snakes have been studied in the Pine Bar-

rens of New Jersey for over 35 years, including marking

each snake with an individual PIT tag (Burger and Zap-

palorti 2011). They are large constrictors that reach the

northern limit of their range in southern New Jersey. The

New Jersey population is separated from other popula-

tions by several hundred km (Golden et al. 2009; Burger

and Zappalorti 2011, 2016). They dig their own nest bur-

rows, and dig or modify hibernacula (Burger and Zappal-

orti 2011). They hibernate communally, along with other

snake species.

Northern Pine Snake studies have examined breed-

ing and hibernation biology, risks and threats Northern

Pine Snakes face, habitat selection, movement and home

ranges, and contaminant exposure. While movement has

been studied throughout the year with the use of radio-

tracking, these previous studies did not provide detailed

movement of the community of snakes using hibernacula

(Burger and Zappalorti 2011). During this period North-

ern Pine Snakes were studied in Burlington, Cumber-

land, and Ocean counties, however, the exact locations

of the studies were not disclosed because of the very high

risk of poaching of Northern Pine Snakes (Burger et al.

2017, 2018).

The present study used passive continuous recording

devices on snakes as they left and entered the hibernac-

ula. In 2017 one hibernaculum was monitored to test the

feasibility of using this tracking method, and in 2018, five

hibernacula were monitored (four units monitored snakes

for the entire year in Bass River State Forest, and one

additional site was only monitored in the fall in Berkeley

Township, Ocean County, known as “Davenport Den’).

Any tagged snake passing by, entering, or leaving one

of these monitored hibernacula was recorded. Data were

generally down-loaded every 2—3 weeks throughout the

Amphib. Reptile Conserv.

year. A recording device was placed at the entrance of

each hibernaculum, and buried so it was not visible (Fig.

1). The device used was the AVID Power TracKer VIII,

a multi-mode reader with memory for PIT tags, made by

AVID Identification Systems, Inc., in Norco, California.

A TracKer unit was placed at each hibernaculum entrance

and covered with a | cm layer of sand to prevent vandal-

ism. The unit has a 6-inch coil reader with leads that can

be up to 4 m long and lead to a device that can record and

store up to 2,500 events, recording the PIT Tag, the time

of day, and the date for each event. The power source was

a 12-volt marine battery. The recorder and battery were

placed in a plastic box, covered with a board for stability

(and to prevent collapse if someone stepped on it by ac-

cident), buried beneath 10 cm of soil, and covered with

leaves and twigs for camouflage (Fig. 1). None of the

equipment was visible on the surface to prevent injury of

the snakes, theft, or vandalism. The technology requires

that snakes are fitted with PIT tags. Although the record-

ers were operating all year, this report only examines

snake activity from 15 October to 31 December 2017 (at

one hibernaculum) and from 1 October to 31 December

2018 (at five hibernacula).

The soil surface temperatures were recorded continu-

ously, all year, near one of the hibernaculum entrances at

Bass River State Forest using an Elitech RC-SUSB Tem-

perature Data Logger. The device was placed in a plastic

bag and covered with a 1 cm layer of sand and moss to

disguise its location. In previous studies, this small re-

corder has worked for well over a year on its original

battery.

This study was only possible because: 1) Many North-

ern Pine Snakes use the same traditional hibernation and

nesting sites (Burger and Zappalorti 2011; Burger et al.

2012; Zappalorti et al. 2014), 2) Gravid female Northern

Pine Snake use the same hibernating and nesting sites

(Burger and Zappalorti 1992, 2015), 3) Hatchlings can

November 2019 | Volume 13 | Number 2 | e191

Burger

2017

All activity

Den 1

Temperature °C

“Oi, “OO, Oe

Date

Temperature °C

Reader Down

012007546

841005587

020795281

840890092

012261123

020585572

021079007

021285039

0213009019

021381340

021836516

014534620

021542803

021608772

021631878

841019086

014534324

020891883

Fig. 2. All activity of snakes at den 1 (Bass River State Forest) in 2017 as a function of date and soil surface temperature. The

colored markers indicated by 9-digit numbers in the legend represent individually tagged snakes. Data for snakes during the period

from 29 October to 11 November (red line) were not recorded because the maximum number of data points the receiver could store

was reached.

be easily found at nesting areas, fitted with PIT tags, and

then followed at the hibernacula, and 4) Therefore, most

of the individual snakes using a given hibernation site are

PIT-tagged. Further, the sexes and ages of all snakes were

known because they had been followed since they were

hatchlings or two-to-three years old. The hibernacula in

this study had been studied for over 30 years, and were

well-known to the snakes and the researchers (Burger

and Zappalorti 2011; Burger et al. 2012). For each snake,

the first reading hit of the season was assumed to be its

initial entry.

Analyses included calculating frequencies, percent-

ages, means, and standard deviations for various be-

havior parameters of the Northern Pine Snake around

hibernation sites in the fall. Data were analyzed using

standard SAS software (Statistical Analysis Systems,

Cary, North Carolina, USA), including Kruskal-Wallis

One-Way Analysis of Variance (ANOVA), with 95%

confidence intervals. Kendall tau was used to determine

correlations among ambient soil surface temperatures

and the temperatures at which snakes exhibited activity.

The best models for explaining variations in snake activ-

ity as a function of temperature and date were developed

using SAS (ProcGLM) procedures. Variables included

were den, year, age, sex, date (only for the temperature

model), and maximum and minimum daily sand surface

temperatures.

Amphib. Reptile Conserv.

Results

Date and temperature: The two primary factors that

might account for entry of Northern Pine Snakes into hi-

bernacula in the fall are date and ambient temperature.

The factors entering the best model (F = 34, P < 0.0001,

r’ = 76) for Julian date of activity (e.g., entering/leaving)

were maximum daytime sand temperature (P < 0.001),

den number (P < 0.04), and perhaps age (P < 0.08). The

factors entering the best model (F' = 122, P < 0.0001, r?

= 90) explaining variation in the temperature of snake

activity (1.e., soil surface temperature when a snake en-

tered, left, or passed by the hibernaculum) were maxi-

mum daytime sand temperature (P < 0.001), minimum

nighttime temperature (P < 0.0001), and sex (P < 0.04).

These factors are explored in greater detail below.

Seasonal activity patterns: While there was virtually

no activity around the hibernacula in August or Sep-

tember, by early October snakes returned to the vicinity

of the dens and began passing by, entering, and leaving

hibernacula. In 2017, the equipment was only deployed

in mid-October (when activity was expected to begin) at

Bass River State Forest, but many snakes were already

active around den | (Fig. 2). In the first year the reader

was initially placed 0.3 m down the tunnel to the hiber-

naculum, and this resulted in one snake sitting in the en-

trance, and running the recorder until it reached the max-

105 November 2019 | Volume 13 | Number 2 | e191

Pituophis melanoleucus vulnerability during fall ingress

Temperature °C

Temp

021804062

845090777

012261123

012077546

021608772

841017858

20795281

841004858

021626044

013091070

014534620

011564817

021080090

021542803

021791327

841007010

014534324

021280836

014843117

= 5 i} | | I }

FEA

“Q. “B. ‘0. ‘o “By. “ y “D. “y. “D. “y. her Le 47, CL aa oh tin oh, td, CL for 014263843

fy. % a f; 4, iG So Ry Yo O- g “> v4 fp 8s? — ? . ce

i, <7) : A) ») “3 by 2, pce we &) E> ty Sy 7) Qy “3 Je S. ») 4 > “ a) 25 7) 845090803

~ ~ ~~ ~ ~ “f/f + ~ ~ ~ ~ ka “4 “fy ~ ~ ool C mel

“p tp “ip “Oz. “Oz, ~Op, “Op, “Or, “Or, “Op, “Gp, “tp “dp “Up “Oz, “Op, “Oy, “Op, “Oz, “Op, “Op, “tp

Date

021285039

021381340

836521818

845091087

Fig. 3. All activity of snakes at four dens in Bass River State Forest in 2018 as a function of date and soil surface temperature.

imum number of data points it could store (n = 2,500).

Figure 2 also indicates the period when the recorder was

not recording. Because of this, the recorder was moved

up to the entrance on 9 November 2017 (so that even if a

snake was in the tunnel, watching the outside world with

its head at the entrance, it would not record the activity

more than a few times). During this down period from 29

October to 9 November some snakes entered (and may

have left) without being recorded. Even so, the pattern

clearly shows 18 different snakes (ages 0 [hatchling]

to 16 years) entering, leaving, and re-entering from 16

October to 17 November 2017. The recorder continued

monitoring through December but due to freezing sur-

face temperatures, there was no more activity.

In 2018, the activity of 25 Northern Pine Snakes be-

gan on 2 October and continued to 7 December at Bass

River State Forest (Fig. 3). The pattern was similar to that

in 2017 in that there was daily temperature variation, and

the snakes entered and left numerous times. It is, how-

ever, important to acknowledge these patterns because

they show that activity is rather constant. Thus, the first

hypothesis of a restricted time period of activity around

the hibernacula was rejected.

Additionally, the Davenport Den (Ocean County) was

monitored in the Fall of 2018. In winter of 2017-2018

only one two-year old Northern Pine Snake and two Corn

Snakes Elaphe guttata (now Pantherophis guttatus) used

this hibernaculum. In the fall of 2018, it was used by two

hatchlings and the same two-year old. There were thus

Amphib. Reptile Conserv.

106

no large snakes that might influence the activity of the

small Northern Pine Snakes; and the hatchlings were ex-

tremely active. The activity at this hibernaculum started

on 1 November and ended on 28 December 2018. Since

there were so few snakes at this hibernaculum, the in-

dividual activity patterns of the hatchlings are given in

greater detail below.

Time of Day: As might be expected for ectothermic spe-

cies, snakes were most active during the day and more

so on warm sunny days. Most activity occurred between

1000 and 1700 h in both years (Fig. 4). In 2017, 77% of

the activity occurred between 1000 and 1600 h; in 2018,

84% of the activity occurred in this same time period.

However, in 2017 one snake left at 2000 h, and in 2018,

one entered at 2045 h, and another entered at 0100 h at

night. These rarely observed nocturnal activities occurred

during relatively high temperatures (> 14° C).

Temperature effects: The activity of the snakes was

plotted against the soil surface temperatures for 2017 and

2018 as a function of date (Figs. 2 and 3). In both years,

the October surface temperatures were high, and they

generally decreased throughout the fall. Snakes did not

enter or leave at the lowest daily temperature (at night),

but sometimes entered or left at the highest temperatures

for the day. Most of the activity occurred at temperatures

of 10° C or above. In both years there was little activity

when the surface temperature fell below 8° C at night.

November 2019 | Volume 13 | Number 2 | e191

Burger

Bass River State Forest

Fall 2017 (1 den)

aa 4

é aé

7 o* tye ma 1

eo e

reg é A e

10 @ Final Winter Entry

e@ Entering

4 Leaving

/ Z / f

a7) “Op *ay °0 ®a “Og “Op

@ Final Winter Entry

Surface Soil Temperature (Celsius)

Pr e Entering

25 me ~" 5% aos 4 Leaving

a ¢

20 a « Passed Entrance

e 4 @a

15, @ as é a ~ f

‘4 ren Ke 7

a P| ny aan

10 .45 a

= 8

5 r=0.1

Ga, Ba, “Oy 2a, “Lo, ‘6, op 18. Op <0. Qn <2. Op Vy,

Time of Day

Fig. 4. Fall activity of snakes in Fall 2017 and Fall 2018 as a

function of time of day and surface soil temperature. Activity

type 1s noted by each symbol.

However, it is noteworthy that for both years, snakes en-

tered or left the hibernacula, even after prolonged periods

of daily low temperatures that reached 0° C in 2017, and

even -5° C in 2018 (Figs. 2 and 3).

Although the daily high and low temperatures and

snake movements were correlated (7 = 0.54 in 2017; 0.51

in 2018; both P < 0.0001), the daily high and low tempera-

tures for each day were more highly correlated (r = 0.71

in 2017; 0.79 in 2018, Fig. 5). Figure 5 indicates when

snakes either entered or left a hibernaculum, or made their

final entry for the winter. Note that some snakes entered at

the same temperature point, and so there are fewer points

than snakes. There are fewer points in 2017 because only

one hibernaculum was monitored; while the 2018 data re-

fer to all four hibernacula at Bass River State Forest. Final

entries were usually at lower temperatures than other ac-

tivities (Fig. 5).

Individual behavior: Figures 2—3 indicate frequent ac-

tivity at hibernacula; individual snakes typically entered

and left more than once (rejecting the initial hypothesis).

The activity of individual snakes was examined only for

2018; individual activity in 2017 was not examined be-

cause equipment failure for a short period made it impos-

sible to know whether any snakes left or entered during

that period. Some snakes entered a den and remained for

the winter (32%), but most did not (68%). At the Bass

River study site, some snakes visited all four of the moni-

tored hibernacula on the same day, often returning to the

first one they entered. Snakes entered or left hibernacula

Amphib. Reptile Conserv.

Out

Final

Entry

Previous Night Low Temperature (Celsius)

Early Sept

Ne 4 a -

Out

xe)

Final

Entry

Previous Night Low Temperature (Celsius)

Pass

“10 15 20 25 30 35

Max Daytime Temperature (Celsius)

Fig. 5. Activity of snakes in Fall 2017 and Fall 2018 as a func-

tion of the maximum daytime temperature and the previous

night’s low temperature.

an average of 5.6 + 0.7 times, switched dens an average

of 1.4 + 0.3 times, and visited 1.8 + 0.2 dens at Bass

River State Forest in 2018. Movement was a function of

age: older snakes moved more often than younger ones

(Table 1). At Bass River State Forest, hatchlings moved

an average of only 2.3 + 1.3 times (Table 1).

However, at the Davenport den, where there were only

two hatchlings and one 2-year old, the movement pattern

was very different. Hatchlings used the hibernaculum as

a home base, and went in and out many times before final

entry. Sometimes they remained near the entrance, but

they mainly moved a few meters away (e.g., the hatch-

lings were not immediately located). The seasonal pat-

terns of the two hatchlings are shown in Fig. 6. The two

hatchlings moved 48 and 66 times, while the 2-year old

moved only 16 times. The lack of older, larger Northern

Pine Snakes at the den may have allowed the hatchlings

to move more freely.

Activity around each den (entering, leaving) varied

significantly (X? = 98, P < 0.0001), and the percentage of

hits at each den varied: den 1 = 25%, den 2 = 59%, den 4

= 12% and den 5 = 4% of total activity at Bass River. The

total activity around the four dens at Bass River State

Forest was 139 hits for 25 snakes. At the Davenport den

November 2019 | Volume 13 | Number 2 | e191

Pituophis melanoleucus vulnerability during fall ingress

Table 1. Movement of Northern Pine Snakes among four monitored hibernacula at Bass River State Forest, New Jersey, USA, in 2018.

Age of Snake (yr)

0-1 2-3 5-7 Over 7 ».G

Any Reading

mean 2.3 1.3 2.8+0.9 6.8+2.5 7140.8 8.7 (0.03)

min/max 1/6 1/5 1/11 Srl

Den Switches

mean 0.3403 0.5+0.5 135207 2.0+0.4 8.0 (0.05)

min/max 0/1 0/2 0/3 0/5

Dens Visited

mean 1.3403 1.3+40.3 1840.3 220.2 7.9 (0.05)

min/max 1/2 1/2 1/2 1/4

845*090* 639 ——— Temperature C

—*e— Snake Activity

Temperature °C

845*090*603

Temperature °C

a Lae “Le. Ly, Pg 2s LR; 1) Ones Ons Ss

24 gy, 2b, 20, 04,07 “07, gb, 20, °0, 0, yO

PILE OP PR FE Ly PN a EEE GE Me tee

Date

Fig. 6. Activity of two hatchlings (tag numbers 845090639 and 845090603) in the fall of 2018 at den (Davenport) as a function of

date and soil surface temperature.

Amphib. Reptile Conserv. 108 November 2019 | Volume 13 | Number 2 | e191

Burger

HIBERNATE

(leave in April)

Rest, Bask Forage Reproduction

Seek Mate

Mate

ge ci bees

oe Female

Dig Nest

¢o coe? Lay Eggs

60 days |

Eggs Hatch

Forage

Hatchlings

Emerge

Enter Hibernacula

(Oct-Dec)

Fig. 7. Schematic of life cycle of Northern Pine Snakes, in-

dicating periods of high vulnerability to human disturbances,

such as fire, off-road vehicles, hunting, and poaching.

it was 120 hits for only three snakes, and the two hatch-

lings accounted for most of this activity.

Discussion

Methodological issues and using the Power TracKer

VIII: Any study of animals in the wild is fraught with

variability and uncertainties in the methods used, in en-

vironmental variation, and in the behavior and ecology

of the species. Data from 2017 indicated that the PIT-tag

recording devices could be used in the field with 12-volt

marine batteries, since they operated properly, and the

data could be retrieved. However, the main problem en-

countered initially related to placement of the receiver

— when it was partway down the hibernaculum entrance,

it recorded continuously as some snakes simply rested in

the tunnel, peering out of the entrance and filling all the

available data points. When the coil was moved to the

front of the entrance, this problem did not exist, but then

it was difficult to determine if a snake merely passed by

the entrance, or entered. This issue could be partly man-

aged by seeing where the individually marked Northern

Pine Snake turned up next. The main difficulty with the

Power TracKer was that the batteries need to be changed

to allow charging every 2—3 weeks depending upon tem-

perature (battery life was shorter at cold temperatures).

Batteries need to be charged on the “slow setting” rather

than the “rapid method;” as the former provided a longer-

lasting charge in the field. Bad weather, heavy rains and

snow, and downed trees from severe storms made getting

to the study site to change the (20 kg!) batteries every

couple of weeks very challenging.

Poaching of Northern Pine Snakes is known to be a

major threat (Burger and Zappalorti 2016), so I opted not

Amphib. Reptile Conserv.

to use solar power or place cameras that might call atten-

tion to the den entrance. Visible solar panels would alert

poachers or vandals to the exact location of hibernacula

entrances, and would also encourage theft. Protecting

the equipment is important since each set-up costs about

$3,000 for the TracKer, leads, batteries, plastic case for

the recorder and batteries, and a wood cover to prevent

excessive rain from entering the plastic case. The con-

tinuously recording thermometers were only $25, and

the manufacturer’s battery lasted at least a year. Lastly,

the tracker should be put in place when the snakes are

underground to ensure that they leave a scent trail when

they leave so that other snakes (particularly hatchlings)

can find the entrance.

With any field study there are weather-related and

other environmental variables that can influence the

behavior of the snakes. Exceptionally warm weather in

2018 resulted in an extended period of ingress into the

hibernacula. No snake entered or left den 1 in 2017 (the

only den monitored that year) after 17 November, but in

2018 snakes continued to move in and out of the five

dens monitored into late December.

Finally, there are uncertainties that relate to the behav-

ior and ecology of the snakes. These uncertainties were

related to age, sex, and individual responses. Age clearly

entered as a factor in explaining the observed Northern

Pine Snake behavior, but this would not have been clear

if the snakes were not of known ages. The behavior of

hatchlings varied depending upon the composition of

the hibernaculum community (see below). Hatchlings

moved very little when they were part of a community

that included snakes of different ages (and sizes), but

hatchlings moved often when there were no larger (older)

snakes present.

Activity patterns around hibernacula in the fall:

Northern Pine Snakes in the present study were very ac-

tive around the hibernacula for over two months. At Bass

River State Forest, snakes moved in and out an average

of six times, often switching dens. The hypotheses that

activity around a hibernaculum was restricted in time,

and that snakes entered a hibernaculum and stayed were

both rejected. Individuals moved from den to den over a

few days or a few weeks, partly dependent upon weather.

That Northern Pine Snakes left a given den and did not

enter another den for several days, but then returned, in-

dicates that there are some other suitable places to shelter

when the temperatures drop at night. Nonetheless, the

snakes came back to one of their original dens when the

weather warmed up enough to move.

The factors that played a role in activity were sea-

son (date), temperature (daytime high and nighttime

low), age, and hibernaculum number. The Northern Pine

Snakes appeared to prefer two of the dens over the oth-

ers. However, even snakes that first entered one of the

“preferred dens” moved to other dens before returning.

One might predict that older snakes, aware of the advan-

tages and disadvantages of one den over another, might

November 2019 | Volume 13 | Number 2 | e191

Pituophis melanoleucus vulnerability during fall ingress

move less than young snakes that are less familiar with

their environment and den options. This, however, was

not the case. The reason for increased switching with in-

creasing age 1s unclear. However, in the absence of larger

(older) snakes, the two hatchlings at the Davenport den

(an isolated hibernaculum) used it as a home base, and

moved in and out frequently, depending upon tempera-

ture. These two hatchlings did not finally enter for the

winter until 28 December.

Factors affecting entry into hibernacula: Clearly

there are seasonal and temperature effects; Northern

Pine Snakes enter hibernacula to avoid freezing winter

temperatures, as do other snakes in northern climates.

Although other studies have examined the temperatures

of snakes during hibernation (Costanzo 1986; Hein and

Guyer 2009), or emergence in the spring (Todd et al.

2009), little is known about the temperatures at which

snakes enter hibernacula in the fall. To study fall behav-

ior requires: 1) individually marked snakes, 2) a method

of recording each snake’s entrance in the fall (date, time

of day), and 3) devices to continuously record the soil

surface temperature to capture the temperature when

snakes enter. A long-term study was required for the first

criterion, and recent technological developments were

required for the latter two. The development of this new

technology makes it possible to more accurately examine

both the seasonal and temperature influences on snakes

entering and emerging from hibernacula, as well as indi-

cating the degree to which snakes move in and out dur-

ing both entry and emergence, before finally dispersing

in the spring to forage, mate, and nest.

In this study, date and sand surface temperature de-

termined when snakes entered and left hibernacula, and

snakes moved an average of about six times before set-

tling in for the winter. Three other factors seemed to also

affect movement: den number, age of the snakes, and for

hatchlings, the presence of older (larger) snakes. Snakes

clearly preferred two of the monitored dens over the oth-

er two, and both of the preferred dens were deeper than

the other two, and they were older in terms of usage his-

tory (Burger and Zappalorti 2011).

The age of the snakes influenced their movement;

older snakes moved more often, switched dens more

often, and visited more dens than did younger snakes.

This was unexpected since younger snakes might be ex-

pected to explore a range of different sites before settling

down. Hatchlings using the four hibernacula at the Bass

River State Forest showed significantly less activity than

older snakes (an average of only two movements/snake).

However, the two hatchlings that used the Davenport hi-

bernaculum showed activity 48 and 66 times. They not

only entered and left (and were not visible around the

hibernaculum or in the surrounding area), but sometimes

basked very near the entrance, moving swiftly down the

entrance when approached by the researchers. They ap-

peared to be using the hibernaculum as a refuge and an

Amphib. Reptile Conserv.

overnight site for nearly two months before remaining for

the winter (refer to Fig. 6). This difference in hatchling

behavior at the two sites may relate to the relative risk

posed by much larger snakes using the same hibernacu-

lum. If large, heavy (up to 1,350 g) adult snakes are en-

tering and leaving, they pose a risk to hatchlings (30—50

g), and adults could injure them while both are moving

through the tunnels. The only dead snakes found tn hiber-

nacula over the years (with one exception of small mam-

mal predation) were those squashed flat by older, larger

snakes lying on top of them for long periods of time.

Vulnerability, risk, and competing claims: Northern

Pine Snakes are most vulnerable when they are roaming

above ground (even though they are partially fossorial),

and when they are concentrated in one small area. They

are above ground at intermediate temperatures; 1n the hot

summer they spend a great deal of time in hollow fallen

logs, under leaves and needles, or underground; in the

winter they hibernate 1—2 m below ground. Behaviorally

they are vulnerable when they are mating (spatially scat-

tered), nesting (females, spatially clumped), and entering

or leaving hibernacula (clumped around hibernacula).

The vulnerability of Northern Pine Snakes is greatest

when these two features (above ground and clumped)

overlap, which occurs when entering hibernacula for the

winter. This situation occurs when entering hibernacula

for all Northern Pine Snakes, and for females when they

are nesting (Burger and Zappalorti 1992, 2011, 2016;

Burger et al. 2017, 2018, Fig. 7). The data presented in

this paper clearly show that the period of ingress into hi-

bernacula is at least two months in duration, spanning

both October and November, and can extend through

December if the weather is not too cold. The data also

show that there is frequent activity, not just one entry into

the hibernaculum by each snake. The dens at Bass River

State Forest that were studied are about 30-120 m from

each other. That snakes come and go indicates that the

spatial area of activity is greater than just around the 1m-

mediate entrance to a hibernaculum.

Only once was a Northern Pine Snake seen above

ground in the fall, although later analysis of the recorded

data indicated that just minutes before or after our pres-

ence, snakes entered or left the hibernaculum. In one

case, a snake came up five minutes after we finished

downloading the data, and I only saw it because I went

back to pick up a piece of equipment. It was lying in the

tunnel, with its head about 2 cm from the entrance. This

observation emphasizes the importance of having con-

tinuously recording equipment; observation alone would

not yield this key information.

The major risks that Northern Pine Snakes face are

natural predators (hawks, mammals, and other ophi-

ophagus snake species), commensal predators (dogs,

raccoons), poachers, loss of habitat, and human distur-

bance (direct and indirect). Human disturbance can take

the form of people disrupting snake behavior (e.g., dur-

November 2019 | Volume 13 | Number 2 | e191

Burger

ing snake copulation, nesting, or entering/leaving hiber-

nacula), disrupting snake habitat (e.g., off-road vehicles,

ORV races through the Pine Barrens, prescribed burns,

tree-cutting), or a combination of these activities. Many

of these activities represent competing claims for the

same Pine Barrens habitat (either quantity or quality of

the habitat).

The resolution of competing claims for Pine Barrens

habitat, or management of that habitat, 1s partly a societal

decision. However, the specific needs of wildlife, and of

specific species such as the Northern Pine Snake, cannot

be considered unless there are data to show what those

needs are, when (and what) their vulnerabilities are, and

when interference will jeopardize their populations. In

many cases, a situation can be resolved by bringing to-

gether the relevant people and agencies, and determining

the best course of action. Clearly, debris and dry leaves

that will result in a hot fire that could destroy local hu-

man communities or businesses (as well as wildlife) need

to be reduced, and fire management is a reasonable op-

tion. Likewise, hunting, ORV races, and other human ac-

tivities are reasonable uses of public forests such as the

Pine Barrens. Protection of endangered and threatened

species is another public goal (as well as being a legal

one) that must be considered. Even accepting the latter

as an important public goal does not completely solve the

problem, however, because different species may have

different requirements and vulnerable periods. For each

species, relative abundance and total distribution need to

be considered, with species which have very restricted

ranges getting priority treatment. A consensus needs to

be reached both about the specific habitat and ecologi-

cal requirements of different endangered, threatened, or

otherwise vulnerable species, and about the specific re-

quirements of other groups with competing claims (e.g.,

foresters, recreationists, fire managers). Armed with this

knowledge, managers can make science-based, societal-

ly-based, and cost-effective decisions about managing

the habitat and the associated wildlife species that occur

there.

Conclusions

Plants and animals in the Pine Barrens, and everywhere

else, face competing claims to their habitat along with

of the risks of disruption or disturbances from other ani-

mals, poachers, recreationists, foresters, resource manag-

ers, firemen, developers, and the general public. Resolv-

ing competing claims requires having knowledge of the

specific needs of vulnerable species and habitats, as well

as the needs of the people and managers. One high risk

vulnerable period for Northern Pine Snakes is when they

enter or leave their winter hibernation sites. This paper

provides data showing that the fall ingress period to hi-

bernacula is prolonged (over two months), and involves

frequent snake activity above ground. During October

and November in the Pine Barrens, Northern Pine Snakes

Amphib. Reptile Conserv.

are moving toward hibernacula, concentrating there, and

entering and leaving frequently until they eventually stay

underground for the winter. Thus, this is a highly vulner-

able period when snakes are concentrated, and any dis-

ruptions (such as fires or off-road vehicle races) have the

potential to injure or kill Northern Pine Snakes that are

threatened in New Jersey, and threatened or endangered

throughout most of their range. New Jersey has perhaps

the largest, and most stable population of Northern Pine

Snakes throughout the range of the species (Golden et al.

2009; Burger et al. 2016, 2017), therefore state wildlife

agencies have a special responsibility to ensure its con-

tinued survival.

Acknowledgments.—I especially thank R.T. Zappalorti

and M. Gochfeld who have been part of these Northern

Pine Snake studies since the beginning, and E. DeVito,

who quickly joined us. I thank the many agencies and in-

dividuals who have helped study and preserve Northern

Pine Snakes in the New Jersey Pine Barrens, especially

Kris Schantz, David Jenkins, and Dave Golden of the

Endangered and Nongame Species Program, and Cyn-

thia Coritz of the Division of Parks and Forestry of the

New Jersey Department of Environmental Protection,

New Jersey Conservation Foundation, and Nature Con-

servancy. Over the years many Rutgers University ecol-

ogy graduate students and personnel of Herpetological

Associates have assisted in these studies. I particularly

thank Christian Jeitner, Kelly Ng, Matt McCort, David

Schneider, Mike Torocco, Dave Burkett, Ryan Fitzger-

ald, and Taryn Pittfield. This research was performed

under Rutgers University Protocol number E6—017, and

appropriate state permits. Funding has included sup-

port from Rutgers University, Herpetological Associ-

ates, New Jersey Conservation Foundation, and the Tiko

Fund, and I gratefully thank the many volunteers who

have cheerfully helped us throughout the 30+ years of the

study, including the teenagers who grew up to continue

helping, bringing their own children.

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Amphib. Reptile Conserv.

Pituophis melanoleucus vulnerability during fall ingress

Joanna Burger is a Distinguished Professor of Biology at Rutgers University, as well

as a member of the School of Public Health, Institute for Marine and Coastal Sciences,

the Biodiversity Center, and the Environmental and Occupational Health Sciences

Institute. Joanna received her B.S. in Biology from the State University of New York at

Albany, her M.S. in Zoology and Science Education from Cornell University, her Ph.D.

in Ecology and Behavioral Biology at the University of Minnesota in Minneapolis,

Minnesota, and an honorary Ph.D. from University of Alaska. She is an ecologist,

human ecologist, behavioral biologist, and ecotoxicologist who has worked with several

species, including Pine Snakes, lizards, turtles, and sea turtles, for over 40 years in

many parts of the world. Joanna’s primary research has focused on behavioral ecology,

ecotoxicology, risk assessment, and biomonitoring. Additional research involves public

perceptions and attitudes, inclusion of stakeholders in solving environmental problems,

and the efficacy of conducting stakeholder-driven and stakeholder-collaborative

research. She has been a member of the Endangered and Nongame Species Council

since the mid-1970s, and has served on several National Academy of Sciences boards

and committees. Joanna has published extensively in the peer-reviewed literature, and

has written or edited over 25 books, including The Northern Pine Snake: Its Life History,

Behavior, and Conservation with R.T. Zappalorti.

114 November 2019 | Volume 13 | Number 2 | e191

Official journal website:

amphibian-reptile-conservation.org

Amphibian & Reptile Conservation

13(2) [General Section]: 115-125 (e192).

Teptile-cons™

From incidental findings to systematic discovery:

locating and monitoring a new population of the

endangered Harlequin Toad

1*Andrés Jiménez-Monge, ‘”*Felipe Montoya-Greenheck, ‘Federico Bolafos,

and °*Gilbert Alvarado

'Faculty of Environmental Studies, York University, Toronto, CANADA *Escuela de Antropologia, Universidad de Costa Rica, San José, COSTA

RICA 3Observatorio del Desarrollo, Universidad de Costa Rica, San José, COSTA RICA *Escuela de Biologia, Universidad de Costa Rica, San

Pedro Montes de Oca, San José, COSTA RICA *Laboratorio de Patologia Comparada de Animais Selvagens, Universidade de SGo Paulo, Sdo

Paulo, BRAZIL °Laboratorio de Patologia Experimental y Comparativa (LAPECOM), Escuela de Biologia, Universidad de Costa Rica, San José,

COSTA RICA

Abstract.—The scientific and conservation communities recognize toads of genus Atelopus as among the

most vulnerable of all amphibian groups, with over 75% of the species assessed as “Critically Endangered” by

the International Union for the Conservation of Nature (IUCN) Red List. Atelopus varius, known as the Harlequin

Toad, has been thought to be extinct in Costa Rica since the mid-1990s. There have been four rediscovered

populations of the species since 2004. This report presents the fifth reappearance of A. varius, this time in the

Alexander Skutch Biological Corridor (ASBC) in the Pacific Slope foothills of La Amistad International Park,

Costa Rica, which represents a new location. Previously, the pattern of reappearance of this species has been

unclear. In this study, the discovery of a new population of A. varius allows us to evaluate the presence of Bd

infection and offer critical natural history remarks. In total, 25 different individuals were identified. All samples

analyzed for Bd diagnosis were negative. In contrast to other A. varius populations, this one was mostly found

high above the riverbed, often in the foliage, tree trunks, and bromeliads, from 1-6 m above the water both

during day and night. The absence of Bd infection in these Harlequin Toads, a highly susceptible species, in

an area identified as having a high probability of Bd occurrence, suggests that this behavior could have helped

this population survive by reducing infection risk. Moreover, the distribution of A. varius may have changed

in the last 50 years, by penetrating higher in the montane regions of the Talamanca mountains, a change in

distribution that might also help its survival of some environmental stressors. With the discovery of a new

locality for A. varius, this study offers an animal behavior argument to account for species recovery in general,

as well as a possible expansion of what has been accepted as the historical distribution of this species.

Keywords. Ate/opus varius, natural history, citizen science, endangered species, chytrid fungus, Alexander Skutch

Biological Corridor, Costa Rica

Resumen.—Los sapos del géenero Atelopus son reconocidos como uno de los grupos de anfibios mas

vulnerables, con mas del 75 por ciento de las especies de este géenero evaluadas como “En Peligro Critico”

por la Lista Roja de la Union Internacional para la Conservacion de la Naturaleza. Se pensaba que Atelopus

varius, conocido como la rana Arlequin, se habia extinguido en Costa Rica desde mediados de los anos

noventa. Después de 2004 ha habido cuatro redescubrimientos de la especie. Este informe presenta la quinta

reaparicion de A. varius, esta vez en el Corredor Biologico Alexander Skutch (ASBC) en las estribaciones de la

vertiente del Pacifico del Parque Internacional La Amistad, Costa Rica como una nueva ubicacion. Hasta ahora,

el patron de reaparicion de esta especie no ha sido claro. En este estudio, con el descubrimiento de una nueva

poblacion de A. varius, ofrecemos importantes observaciones de historia natural y evaluamos la presencia

de la infeccion por Bd. En total, se identificaron 25 individuos diferentes. Todas las muestras analizadas para

el diagnostico de Bd fueron negativas. En contraste con otras poblaciones de A. varius, en nuestro caso se

encontro la mayoria de los individuos alto sobre el lecho del rio, a menudo en el follaje, troncos de arboles y

bromelias, entre 1-6 m sobre el agua, tanto de dia como de noche. La ausencia de infeccion por Bd en estas

ranas arlequin, una especie altamente susceptible, en un area identificada como con una alta probabilidad de

ocurrencia de Bd, sugiere que los sapos que pasen menos tiempo cerca del rio y mas tiempo en areas abiertas,

podria haber ayudado a esta poblacion a sobrevivir mediante la reduccion al riesgo de infeccion. Ademas,

hipotetizamos que la distribucion de A. varius pudo haber cambiado en los ultimos 50 anos, penetrando mas

alto en las regiones montanas de la cordillera de Talamanca, un cambio en la distribucion que también podria

Correspondence. ':* andresjmo@gmail.com ORCID: 0000-0002-8787-0820; !?° fmontoya@yorku.ca ORCID: 0000-0002-8273-5515;

‘federico. bolanos@ucr.ac.cr ORCID: 0000-0002-7935-64 18; °° gilbert.alba@gmail.com ORCID: 0000-0001-8418-043X

Amphib. Reptile Conserv. 115 October 2019 | Volume 13 | Number 2 | e192

New Atelopus varius population in Costa Rica

estar ayudando con su supervivencia a ciertos factores ambientales. Con el descubrimiento de una nueva

localidad para A. varius, nuestro estudio ofrece un argumento de comportamiento animal para explicar la

recuperacion de especies, asi como una posible expansion de lo que se ha aceptado como la distribucion

historica de esta especie.

Palabras clave. Ate/opus varius, historia natural, ciencia citudadana, reaparicion de especies amenazadas, rana arle-

quin, hongo quitrido, Corredor Biolégico Alexander Skutch, Costa Rica

Citation: Jiménez-Monge A, Montoya-Greenheck F, Bolafios F, Alvarado G. 2019. From incidental findings to systematic discovery: locating and

monitoring a new population of the endangered Harlequin Toad. Amphibian & Reptile Conservation 13(2): [General Section]: 115-125 (e192).

tribution 4.0 International (CC BY 4.0): https://creativecommons.org/licenses/by/4.0/], which permits unrestricted use, distribution, and reproduction in

any medium, provided the original author and source are credited. The official and authorized publication credit sources, which will be duly enforced,

are as follows: official journal title Amphibian & Reptile Conservation; official journal website: amphibian-reptile-conservation.org.

Received: 14 September 2018; Accepted: 16 July 2019; Published: 23 October 2019

Introduction

Over 75% of the species in genus Ate/opus are assessed

as “Critically Endangered” by the International Union

for the Conservation of Nature Red List (IUCN 2015).

Within this genus, the Harlequin Toad (Ate/opus varius,

in Spanish as “Rana Harlequin”), 1s the last remaining

species in Costa Rica. The other three representatives are

believed to be extinct (Barrio-Amoros and Abarca 2016).

This moderately-sized toad 1s associated with small,

fast-moving streams, where it is found along the banks

and sitting on rocks in the stream. Endemic to Costa

Rica and Panama, at one point there were over 100

known populations in both the Atlantic and Pacific Slope

versants of the mountain ranges in Costa Rica and western

Panama, reaching up to 2,000 m asl (Savage 2002). In

the mid-1990s, A. varius was believed to be extirpated

from Costa Rica, following a drastic decline that began

in Monteverde in 1988, where it remains absent (Pounds

and Crump 1994). Apart from habitat loss, two other

leading possible explanations for the Harlequin Toad’s

disappearance are climate variations and fungal disease.

Research so far suggests the possibility that the decline

of A. varius has a multifactor explanation (Pounds et

al. 2010; Berger et al. 1998) combining both the effect

of climate stress and the appearance of the fungus

Batrachochytrium dendrobatidis (Bd).

Since its loss during the 1990s, four known re-

discoveries of the species have been documented, each

in private properties in the Pacific slope in Puntarenas

Province, Costa Rica. In 2003, a population was

discovered at Fila Chonta, Quepos (Pounds et al. 2010;

Ryan et al. 2005). Nine years later, to the South, a new

breeding population was found near Las Tablas, near

the Panamanian border, at 1,300 m asl (Gonzalez-Maya

et al. 2013). One individual of A. varius was reported

near Buenos Aires, at an elevation of 840 m asl (Solano-

Cascante et al. 2014). Moreover, there was a recent

rediscovery of A. varius by Barrio-Amoros and Abarca

(2016) at 400 m asl in a rocky stream located in the Uvita

Region. Those authors observed nine adult males, one of

which was dead. They tested two live individuals and the

one dead specimen for Bd, and the dead specimen was

the only one to test positive for Bd (Barrio-Amoros and

Abarca 2016).

The pattern of previous reappearances of this species

has been unclear. In some cases, it appeared in small

Amphib. Reptile Conserv.

secluded creeks (Barrio-Amoros and Abarca 2016), and

in others, it appeared in large, open, and fast flowing

rivers (Gonzalez-Maya et al. 2013; Solano-Cascante et

al. 2014), and at altitudinal ranges from almost sea level

to as high as 1,500 m asl. The sites identified thus far

are all on the Pacific slope, amid degraded landscapes

in unprotected land and fragmented, isolated conditions.

This pattern of reappearance makes these populations

highly vulnerable to habitat loss and environmental risks,

such as the population at one of the known localities in

Costa Rica which is under serious threat of landslides

(Pounds et al. 2010).

Here a newly discovered population of A. varius is

presented, found in the Alexander Skutch Biological

Corridor (ASBC), in the Pacific Slope foothills of La

Amistad International Park. We present crucial natural

history remarks and evaluate the presence of Bd infection

in this population. The natural history of amphibian

species can hold important clues for their survival, given

that the effect of fungal infection can be modified by the

host’s ecology, behavior, and life history (Woodhams

et al. 2008). The exact location of the site 1s withheld,

given that the protection of such sensitive information

is of utmost importance in the case of A. varius in Costa

Rica. All the re-discovered populations are outside of

national protected areas and hence are highly susceptible

to commercial trade, collection, and the irresponsible

handling that can lead to a decrease in population

or increased risk of disease. The IUCN guidelines

acknowledge this risk regarding sensitive data access and

the publishing of data that identify specific geographic

locations IUCN 2012).

Materials and Methods

The ASBC extends over 6,012.60 ha (Fig. 1) within the

canton of Pérez Zeledon in the province of San José. The

principal communities of the ASBC include Quizarra,

Santa Elena, Montecarlo, San Francisco, San Ignacio,

Santa Marta, Santa Maria, and Trinidad. The ASBC

lies between 650 and 1,650 m asl and contains the first

remnant population of A. varius outside the province of

Puntarenas.

Note that the findings documented by community

members and students of the Faculty of Environmental

Studies of York University were essential in locating this

population of A. varius. The incidental sightings came

from at least three separate locations along two different

October 2019 | Volume 13 | Number 2 | e192

Jimenez-Monge et al.

GHIRRIPO PAGIFICO

9.400

Key

[) Chirripé National Park

(9) Las Nubes Biological Reserve

GH Alexander Skutch Biological Corridor

GB Los Cusingos Bird Sanctuary

MM Core Corridor

— Rivers

® Towns

Elevation

Ml 500

m™) 1000

1500

2000

Me 2500

QUEBRADA\GHANGHOS

-83.600

Fig. 1 Location and boundaries of the Alexander Skutch Biological Corridor (ASBC) in the province of San José, Costa Rica.

rivers. The rivers are at least 2.3 km apart in a straight

line and separated by mountainous formations. Some

observers had seen toads along the same river, but the

sightings were 300 m apart. In-depth interviews were

conducted with the observers of the Harlequin Toad

incidental sightings to locate the population. After the

interviews, 18 survey days followed, during which over

65 hrs were spent locating, observing, and sampling A.

varius in the previously identified locations.

The interviews aimed to determine as accurately as

possible the location, date, time, available GPS data

(geo-tracking in mobile devices), weather, and relevant

information of each sighting. In April and June 2016,

two visits to the narrowed-down sites were conducted

and systematic searches were performed. These included

six independent diurnal explorations ranging from 3-5

h each, mostly starting at 7:00 AM, with night surveys

also conducted to try to find individuals sleeping in the

foliage. These visits were not successful in locating any

individuals of the population. However, in February

2017, another search successfully located the area

where A. varius resides. After the first individual was

encountered, a transect of 1.1 km was established to

monitor the identified locality systematically.

The transect was surveyed continuously for six days

in February 2017, nine days in June 2017, and three

days in January 2018. These months were selected to

account for seasonal variation, in order to increase the

chances of an encounter. Systematic surveys started early

in the morning and lasted for an average of four h, with

additional surveys carried out sporadically at night to

identify additional locations and sites. The transect was

hiked starting on opposite ends alternatively to account for

possible time variations of activity of the toads. The search

process usually involved 4-6 h of daily effort, walking

along the river and searching in caves, rocks, foliage, and

Amphib. Reptile Conserv.

vegetation above the river. Upon encountering the toads,

they were swabbed for Bd samples, and individuals were

measured and photographed both dorsally and ventrally.

Upon the conclusion of these procedures, individuals

were released in the same location, and each was

handled with a separate set of gloves. Fourteen of these

individuals were sampled for Bd, 10 during the February

2017 surveys and four during June 2017, two of which

were juveniles.

Sex and age were also recorded, along with substrate,

activity (Rest/sleep, Hide, Bask/Splash, Walk/Climb/

Feed), and distance from the river upon first encounter.

For simplicity, the behavior labelled as “splash” refers to

moisture control, as toads absorb water from wet surfaces

in the stream's splash zone (Pounds and Crump 1994).

For better data analysis, the substrate Vegetation was

classified as either trees, bromeliads, branches, or tree

trunks; Soil/Rock refers to either bare ground, crevices,

boulders, or mossy regions on top of the bare rock. The

distance from the river was recorded with horizontal and

vertical distances measured from the water’s edge to the

perch where the toad was found.

The skin swab and storage protocol by Whitfield et

al. (2017) was used, which involves rubbing a sterilized

swab (MW-100 cotton-tipped swab) on the dorsum,

the venter, the sides, and the limbs. Nucleic acids were

extracted from the swabs using Prepman Ultra and Real-

Time PCR protocols (Boyle et al. 2004; Kriger et al. 2006)

in the Laboratory of Experimental and Comparative

Pathology, School of Biology, University of Costa Rica.

Positive and negative controls were run in triplicate on

each 96-well PCR plate. Bd primers and probes (Boyle

et al. 2004) were used in a TaqMan® Gene Expression

Assay (Applied Biosystems, Carlsbad, California).

Samples were run in an Applied BioSystems Prism 7500

Sequence Detection System in Centro de Investigacion

October 2019 | Volume 13 | Number 2 | e192

New Atelopus varius population in Costa Rica

en Biologia Celular y Molecular (CIBCM), University of

Costa Rica. Samples corresponding to 14 animals from

the study site are described.

Results

Open-ended Interviews of Incidental Sightings

In 2015, two independent incidental sightings of the

Harlequin Toad were documented, photographed, and

reported by local community members of the ASBC to

York University’s LNP Director (FM). In March 2016,

a York University student spotted and photographed

another A. varius specimen in the ASBC. Subsequently,

in August 2016, new sightings of the Harlequin Toad

were further documented by students and community

members. Additional sightings of individuals were also

reported by the Tropical Science Center Los Cusingos

Administrator Mario Mejia for May, August, and

September of 2016. During these two years of incidental

sightings, the authors engaged in open-ended interviews

with the incidental observers to expand on_ their

information. These interviews provided precious data for

reducing the search area and narrowing the times of the

day for targeted searches.

Transect Observations

In February 2017, 13 individuals in a rocky stream

within the identified locations in the ASBC were

photographed and identifying body patterns were noted.

Eleven were males and two were females—no juveniles,

eges, or tadpoles were observed. In June 2017, six new

individuals, identified by their unique body patterns,

were recorded in the same study area—three females

and three juveniles. In August 2017, at least one other

individual was observed. In total, with the transect

observations and the photographed incidental sightings,

at least 25 different individuals have been identified in

these locations. The sites where this population resides

are along a wide, open, and fast flowing river, with a

young riparian forest and some old tall trees, surrounded

by a disturbed landscape, with only 10 to 20 m of riparian

forest habitat in some areas.

Males had an average snout-vent length of 25.7 mm

and females 34.3 mm. Male dorsal patterns appear to

include more yellow and green tones, and females have

a stronger orange and red coloration (see Fig. 2). This

coloration is consistent with the remarks of Savage

(1972) on Atelopus populations of Panama and Costa

Rica. Juveniles, on the other hand, have no red and tend

towards lime green, with patterns that are much more

speckled (Fig. 3C,E). All individuals appeared to be in

good health, and no external lesions were present; in

most cases, the skin was colorful, vibrant, and healthy.

Interestingly, the females had very wrinkly cloaca,

probably due to oviposition; no males exhibited any

similar skin condition. One female was molting (Fig. 2E),

from whom skin samples were collected and observed

under the microscope, which detected no presence of Bd.

In total, 14 individuals were tested for Bd, all of which

tested negative for Bd diagnosis.

Amphib. Reptile Conserv.

Just one-quarter of the identified adults were females,

and males were sometimes observed in groups. There

was no indication of reproduction in February, while

in June, juveniles were located, all of them at least 2 m

above the river and in the foliage. In contrast to other

populations of A. varius, the toads in the ASBC were

mostly located above the riverbed, often found in the

foliage, tree trunks, and bromeliads between 1-6 m

above the water, both during day and night. Of the 37

recorded observations, only 11 individuals were seen

close to the river or the splash area, and 10 of them were

seen in vegetation between 3-6 m high.

Natural History Notes

During the surveys, there was ample opportunity to

observe the natural history of this species, and these

observations can help other scientists better locate and

identify new sites in the field, especially when there are

previously existing reports from community members.

Despite the Harlequin Toad’s bright coloration, this

species can be challenging to locate in the field. For

example, during our systematic exploration, the first

individuals were found after almost 25 h of searching in

the same location where it was later documented.

The start of diurnal activities varied among the

seasons. During the dry season, toads were easily located

very early in the morning; but during the wet season,

toads were less frequent, and surveys needed to start

much later when the day had warmed up. Nineteen

sightings were obtained during January and February, but

only six individuals in June, even though more time and

effort was spent during this month. Overall, the sightings

of females were less abundant, and for both sexes, there

were more individuals seen during the dry season. There

was a change in sex ratio between the two seasons (Fig.

4,G=4.216, 1 df, P=0.040), with proportionately more

males detected during the dry season than the wet season.

Regarding the substrate, females were generally

absent from the leaf litter and mostly found on the

vegetation and river boulders, while males were found

on the leaf litter (Fig. 5, G = 9.035, 2 df, P = 0.011).

Previous records suggest that this toad is most often

found along the banks, sitting on the rocks near the

splash zone (Crump and Pounds 1985; Pounds and

Crump 1994; Savage 2002). However, in this case, adults

were typically found in the vegetation or on the ground,

basking on the rocks or foraging along the river bank,

and rarely seen in the splash zone (Fig. 6). Juveniles had

a stronger preference for vegetation (Fig. 6, G= 8.365, 2

df, P =0.015), which explains why they were so difficult

to locate (Fig. 3E).

The toads were observed basking more during the wet

season, on the rocks near the openings in the canopy or

the exposed vegetation (Fig. 2E-G). For example, the

three females in Fig. 2 are pictured as found: the first was

basking, exposed on the vegetation in an open part of the

river around 9:00 AM during the dry season; the second

and third females were basking on the river rocks during

the wet season. Hiding and basking were activities found

to be significantly associated with the dry season, while

hiding was never seen during the wet season (Fig. 7, G

October 2019 | Volume 13 | Number 2 | e192

Jimenez-Monge et al.

ZA ‘ 1A #

. i f !

' i: "oh

i — "ear. he

’ i ‘

o XL. _— E F ‘ =

Fig. 2 Detail of Atelopus varius individuals found during February and June 2017 in the ASBC. The left column includes males;

the right column includes females. Males (B) and (C) were photographed as found, as were females (E), (G), and (H). Note the

spread-out basking position of female (G).

Amphib. Reptile Conserv. October 2019 | Volume 13 | Number 2 | e192

New Atelopus varius population in Costa Rica

Fig. 3 Males, females, and juveniles of Ate/opus varius found in the Alexander Skutch Biological Corridor. (A) Male found during

cm ag ™

F ho % bing

a r

. a ‘ - =

ee - sg

February surveys. (B) Sleeping male in the leaf litter in the same location of a male in Fig. 2(A). (C) and (E) are juveniles high

above the river bank, at least 3-4 m high in the vegetation of the understory, and juvenile (C) is sleeping. (D) A female Harlequin

Toad sleeping on the vegetation five m above the river.

= 13.159, 1 df, P = 0.004). When rains were abundant,

the toads were found sleeping or resting high on the

vegetation.

A difference in activity patterns was also observed

between males and females of this species (Fig. 8, G

= 15.247, 1 df, P = 0.002). During this study, females

were mostly seen basking on river boulders, or on top

of vegetation above the river; while males were more

passive, with significantly predominant activity in the

leaf litter and crevices of the riverbed, hiding, or resting

(Fig. 3A-B). On the other hand, the activity for juveniles

of this species mostly involved sleeping or active

movement for feeding. Juveniles were never observed

basking or hiding like the adults (Fig. 9, G= 9.934, 1 df,

P=0.019).

The Harlequin Toads studied here seemed to be

very faithful to their sleeping grounds. On five separate

occasions, three individuals, including a juvenile (Fig.

3C), were seen sleeping on the same leaf during the night.

Besides the observed nocturnal feeding behavior, almost

Amphib. Reptile Conserv.

all the individuals seen during the night were sleeping

in the vegetation high above the river. Activity started

before dawn. One of these individuals was monitored

during the night, and its activity started before 4:30 AM.

Toads sleeping high in the vegetation (Fig. 3 C,D) made

their way down to lower levels of the ground as the

morning warmed up, with some of them remaining on

the vegetation during the day (Fig. 2E).

Discussion

Based on Savage’s (1972; 2002) reviews of A. varius

distribution in Costa Rica and Panama, it is clear that

most of the recently re-discovered populations have been

found in areas of their historical distribution. The report

by Ryan et al. (2005) for Fila Chonta, 10 km northwest of

Quepos, matches the premontane distribution suggested

for southwestern Costa Rica, where the nearest site

identified by Savage (1972) is Bart. The account by

Barrio-Amoros and Abarca (2016) for the Uvita region

October 2019 | Volume 13 | Number 2 | e192

Jiménez-Monge et al.

O Female

GO Male

Individuals

Dry Wet

Season

Individuals

Fig. 4 Occurrence of males and females of Atelopus varius according

to season.

DAdult

DJuvenile

2 10

are

= 6

Soil/rock Vegetation Leaflitter

Substrate

Fig. 6 Substrate preference of Ate/opus varius according to age.

Ol Female

8 G Male

& 5

= 4

Rest/sleep Hide Bask/Splash Walk/Climb/

Feed

Activity 7

Fig. 8 Activity pattern of Ate/opus varius according to sex.

matches with that reported by Savage (1972); and finally,

the rediscovery in Las Tablas by Gonzalez-Maya et al.

(2013), brings hope for the survival of the toads near

Coton, also previously identified by Savage (1972).

These findings confirm that some populations in the

Pacific versant have managed to escape the decline while

persisting in the same localities, at least so far. However,

evidence from our research, with the discovery of a new

locality, offers augmented hope for species recovery. We

hypothesize that the behavior of this population could

have helped to reduce the risk of exposure to Bd infection

and also to allow a possible change in the historical

distribution of this species.

These results, and the similarity between both

the scientific and community accounts discussed

below, suggest that A. varius distribution could have

Amphib. Reptile Conserv.

Individuals

OrRFNW HU DN WO WO

Individuals

121

O Female

G Male

Soil/rock Vegetation Leaflitter

Substrate

Fig. 5 Substrate preference of Ate/opus varius according to sex.

pany

io)

O Dry

O Wet

Rest/sleep Hide Bask/Splash Walk/Climb/

Activity Feed

Fig. 7 Activity pattern of Atelopus varius according to season.

O Adult

GB Juvenile

Rest/sleep Hide Bask/Splash Walk/Climb/

Activity eat

Fig. 9 Activity pattern of Atelopus varius according to age.

changed to penetrate higher in the montane regions of

the Talamanca Mountain Range in the Pacific Slope.

Despite the lack of specimens for the area and the lack

of a collection archive, previous evidence has recorded

A. varius, specifically this ecomorph, in the San Isidro

del General region at 704 m asl (Savage 1972), with no

other accounts found by the authors for the ASBC area

(from 1,100—1,500 m asl). Furthermore, Savage (1972)

refers to this toad as a predominantly premontane frog

with penetration into lower montane zones only at six

localities in Costa Rica, none of which includes the

area of this study—the closest area identified is Pérez

Zeledon at 704 m asl (Savage 1972). Taking Savage’s

(1972) account into consideration, the discovery of this

population in montane areas of the ASBC suggests this

is an undocumented locality. This scientific account is

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New Atelopus varius population in Costa Rica

congruent with interviews of highly experienced locals

who do not report or remember this species being present

at elevations higher than 600—700 m asl.

Long-time residents of what is now the Alexander

Skutch Biological Corridor remembered that the

Harlequin Toad had been abundant in the area some

50 years ago, but that this toad had not been reported

again for decades, until now. During initial interviews,

local observers showed good competency to immediately

identify Harlequin Toads. One of them acknowledged the

remarkable altitudinal change for this species over the

years, “this frog was very common at 600 m over twenty

years ago, now it is over 1,000 m. It was never here, and

it has come higher in the mountain. It’s the first time

I see it again in all these years” (Ramon Mora). This

citizen testimony along with Savage’s (1972) account

could point to a possible geographic displacement. When

considering the possible displacement, we acknowledge

that reduced biodiversity exploration in the area could

explain how this population went under the scientific radar

for so long; yet, with no valid reason, this assumption

would disregard the confidence, skills, experience, and

traditional knowledge of the inhabitants that are quite

confident the frog was not present over 700 m asl in the

past. If this species were not so easily identified, with

such a reduced risk of misidentification in the field, we

would consider this possibility. This possible change in

distribution might be helping to facilitate its continuing

survival through some environmental stressors, such as

climate change and habitat loss. It might also increase the

risk of Bd exposure, and clearly more research is needed

to corroborate this possible explanation, exploring the

trade-offs between adaptation to environmental changes

and Bd exposure.

The reappearance of A. varius in this region could

be the result of several possible factors. One hypothesis

is that establishing the biological corridor in 2005, and

subsequent efforts to enhance ecological connectivity

between forest patches, have allowed for the recovery

of habitats that previously served as niches for relict

populations on the verge of extinction, allowing those

populations to recover and relocate, possibly pushed by

environmental changes. Another potential explanation

has to do with the possible emergence of resistance

among these threatened relict populations of A. varius

to the previously devastating Bd fungal disease (Perez

et al. 2014).

To expand on the emergence of resistance to the Bd

infection hypothesis, we present two critical remarks that

contribute to the natural history of the species and might

be related to this population’s survival. First, the toads

in the area are seen less often on the splash region of

the river, and more often on the vegetation, regardless

of seasonality. Second, the typical sedentary behavior

in the mossy rocks near the splash zone associated with

this species (Pounds and Crump 1994) is not as typical in

this location. The toads are not so sedentary, and basking

seems to be more important here than capturing moisture

in the splash zone. Many authors, including Pounds and

Crump (1994), recognize that this species depends on

the moisture of the splash zone and individuals tend to

ageregate in the waterfall splash with the progression

Amphib. Reptile Conserv.

of the dry season. The experiences of working with this

population complement this information. Here, toads

are regularly seen in the foliage, tree trunks, and the leaf

litter, and not on the splash zone, only seven of the 37

registered encounters were found on the river rocks or

crevices. The basking behavior 1s as important, or perhaps

more so, than moisture-seeking behavior. The toads

appear to move to open areas on the river bank or the

exposed vegetation where there is more solar radiation;

and the selection of such areas was more frequent during

the warmest time of the morning, especially on colder

days. This selection could also explain the difference of

activity during dry and wet seasons, where during the

wet season the toads were “lazier” in the early hours of

the morning when it was cold and they were found closer

to the river when the day was warmest. These individuals

were much less sedentary than previously reported,

engaging in significant daily movements. Anurans

sleeping on the vegetation 3-4 m above the river move

during the morning down to the river bank; explaining

why this species has been hard to locate in the area, as

researchers might focus solely on crevices, rocks, and

roots as reported by most of the literature. Understanding

this was the key to finding the juveniles that were far

from the river and much higher than eye-level.

In other words, the natural history remarks noted

above suggest that A. varius spends less time closer to

the river and more time in open areas basking; which

in turn might be linked to available solar radiation and

moisture control. Temperature (Woodhams et al. 2003)

and moisture (Johnson et al. 2003) have been suggested

as two important environmental factors influencing the

growth and survival of B. dendrobatidis. So this behavior

could explain why this toad survived in an area cataloged

as highly likely for the existence of Bd (Puschendorf et

al. 2009). Furthermore, the negative results of the Bd test

support this theory. The documented absence of infection

in these Harlequin Toads, a highly susceptible species in

an area identified with a high probability for occurrence

of Bd, allows us to hypothesize that spending less time

closer to the river and more time in open areas basking,

could help the toads of this population survive by

reducing infection risk. The findings of Woodhams and

colleagues (2003) support our hypothesis; they present

evidence that short periods of high body temperature can

eliminate the pathogen from its hosts, with experiments

suggesting that normal thermoregulation can clear frogs

of chytrid infection (Woodhams et al. 2003). We hope

this account can help provide a better understanding of

the behavior of Ate/opus, as this behavior might hold

clues for the frog’s resistance to Bd or climatic changes,

and can also help to locate new populations.

Conclusions

Garcia-Rodriguez et al. (2012) ask “Where are the

survivors [of] relict populations of endangered frogs

in Costa Rica?” In this research, both community

members and scientists have contributed to answering

this question. Moreover, with A. varius being among the

most endangered of all amphibian species (La Marca et

al. 2005), it is certain that the discovery of the Harlequin

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Jimenez-Monge et al.

Toad in the Alexander Skutch Biological Corridor makes

the conservation efforts in this biological corridor of

critical importance. This population is_ particularly

relevant because of ongoing attempts by private

companies to obtain permits for building a hydroelectric

plant that would dam the two major streams in the ASBC,

destroying the habitats successfully recovered during

a decade of conservation efforts in the corridor. The

reappearance of the Harlequin Toad, currently classified

as Critically Endangered (IUCN 2015), lends supporting

evidence to the effectiveness of this habitat recovery.

The active involvement of citizens in the discovery

of A. varius in the ASBC points to the importance of

strengthening the citizen science component to further

the knowledge base regarding this and other species in the

area. The historical role of citizen science in ecological

research has been generally overlooked, despite its

significant contributions (Miller-Rushing et al. 2012;

Dickinson et al. 2012). The opportunistic and widespread

nature of these sightings suggests that strengthening

citizen science approaches can help to maximize

resources and opportunities for encountering this and

other rare, endangered species. Citizen science creates a

nexus between science and education that expands the

frontiers of ecological research and public engagement

(Newman et al. 2012). After all, without the enthusiastic

and curlosity-filled reports from the community, the re-

emergence of this rare and endangered species might have

gone completely undetected by the scientific community.

Moreover, this participatory process can generate new

meanings and values for herpetofauna in the community,

leading to improved human behaviors directed at the

protection of these and other vulnerable species.

In addition to continuing to monitor for the presence of

A. varius in the ASBC, the community initiative that has

already documented the reappearance of the Harlequin

Toad in the biological corridor will also be useful in

providing further data to help answer pending questions

regarding the impacts of climate change, fungal disease,

and pesticides on these fragile populations.

Acknowledgments.—We thank community members

Hans Homberger, Ramon Mora (“Monchito”), Mario

Mejia, Byron Valverde, Walter Arias, and Christian Arias

for their citizen contributions; and Luis Angel Rojas

for logistical support in the field. We are also grateful

to York University Faculty of Environmental Studies

graduate students Stephanie Butera and Carmen Umafia

for sharing their documented sightings; and to the Fisher

Fund for Neotropical Conservation, for financial support

to the Las Nubes Project. We also thank the private,

anonymous donors that supported the research conducted

by AJM and his graduate degree: your support helped

make this possible. Finally, we would like to recognize

the support of Gerardo (Cachi) Chaves for the creation of

the Alexander Skutch Biological Corridor map presented

in this publication.

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EE, Vargas Quesada J, Sandi Méndez D. 2014. Nota

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Woodhams DC, Alford RA, Marantelli G. 2003.

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October 2019 | Volume 13 | Number 2 | e192

Amphib. Reptile Conserv.

Jimenez-Monge et al.

Andrés Jiménez Monge is a biologist from the Universidad de Costa Rica (San

José, Costa Rica) with a Master’s degree in Environmental Studies, and a Diploma

in Business and Sustainability from York University (Toronto, Ontario, Canada).

Currently, Andrés is working with Bird Studies Canada as Urban Program Coordinator.

His research fields include organizational behavior, change management, conservation,

and herpetology. Andrés strives for creating connections between people and the

planet; find more of his work at http://www.razaverde.com and http://www.udemy.

com/birdwatching.

Felipe Montoya-Greenheck has a B.S. in biology from the University of New Mexico

(Albuquerque, New Mexico, USA), a Master’s degree in tropical ecology from the

Universidad de Costa Rica (San José, Costa Rica), and a Ph.D. from the University

of New Mexico. Currently, Felipe is the director of the Observatorio del Desarrollo,

a research center of the University of Costa Rica, and the director of the Las Nubes

program of the Faculty of Environmental Studies at York University (Toronto, Ontario,

Canada). His research fields include environmental anthropology, rural development,

conservation, and wellbeing.

Gilbert Alvarado is a Costa Rican biologist of the Universidad de Costa Rica (UCR).

Gilbert completed his studies as a veterinarian from Universidad Nacional de Costa

Rica (UNA), and his master's studies in the Regional Postgraduate Program in Biology

of the UCR. He is currently developing his doctoral studies in the Experimental and

Comparative Pathology Program of the Faculty of Veterinary Medicine and Animal

Sciences, University of Sao Paulo, Brazil, in the Laboratory of Wildlife Comparative

Pathology, sponsored by the Office of International Affairs and the UCR. Gilbert was

a professor of Pathology and Anatomy at the UNA, and he has been a professor of the

Section of Zoology and researcher of the School of Biology of the UCR. He has been a

researcher at the Research Center for Microscopic Structures from 2013-2014; as well

as the veterinary adviser of the Biological Testing Laboratory in 2015; both academic

units of the UCR. As principal investigator and collaborator, Gilbert has developed

different projects within his fields of interest (comparative pathology of wildlife, host-

pathogen relationships, amphibian diseases, and conservation of critically endangered

Species), and served as regent and veterinary adviser in herpetofauna centers. He is

founder and coordinator of the Laboratory of Experimental and Comparative Pathology

of the School of Biology, UCR.

Federico Bolafios has done master’s studies at the Postgraduate Regional Program

in Biology in Universidad de Costa Rica. Currently, Federico is a professor and

researcher at the Biology School in the same institution and curator of the Herpetology

Collection of the Zoology Museum. Federico’s research is in the fields of conservation,

systematics, ecology, and behavior of amphibians and reptiles, but his publications deal

mainly with the first topic.

125 October 2019 | Volume 13 | Number 2 | e192

Official journal website:

amphibian-reptile-conservation.org

Amphibian & Reptile Conservation

13(2) [General Section]: 126-132 (e193).

Thorius narismagnus (Amphibia: Plethodontidae):

rediscovery at the type locality and detection of a

new population

‘José L. Aguilar-Lopez, ?*Paulina Garcia-Banuelos, *Eduardo Pineda, and *Sean M. Rovito

'23Red de Biologia y Conservacion de Vertebrados, Instituto de Ecologia, A.C., Carretera antigua a Coatepec 351, El Haya, Xalapa, Veracruz,

MEXICO *Unidad de Genémica Avanzada (Langebio), Centro de Investigacion y de Estudios Avanzados del Instituto Politécnico Nacional, km 9.6

Libramiento Norte Carretera Irapuato-Leon, Irapuato, Guanajuato CP 36824, MEXICO

Abstract.—Of the 42 Critically Endangered species of plethodontid salamanders that occur in Mexico, thirteen

have not been reported in more than ten years. Given the lack of reports since 1976, the minute plethodontid

salamander Thorius narismagnus is widely considered as missing. However, this report describes the

rediscovery of this minute salamander at the type locality (Volcan San Martin), as well as a new locality on Volcan

Santa Marta, 28 km southeast of its previously known distribution, both in the Los Tuxtlas region of Veracruz,

Mexico. The localities where T. narismagnus has been found are mature forests in a community reserve on

Volcan San Martin and a private reserve on Volcan Santa Marta. The presence of maxillary teeth, generally

absent in Thorius, are reported here in some T. narismagnus females. Two efforts which may contribute to the

conservation of Thorius narismagnus are the preservation of the cloud forests where this species persists, as

well as the determination of the presence and possible effect of chytrid fungus in these populations.

Keywords. Conservation, ecological reserve, Los Tuxtlas, missing species, minute salamander, molecular analysis,

Mexico

Citation: Aguilar-L6pez JL, Garcia-Bafuelos P, Pineda E, Rovito SM. 2019. Thorius narismagnus (Amphibia: Plethodontidae): rediscovery at the type

locality and detection of a new population. Amphibian & Reptile Conservation 13(2) [General Section]: 126-132 (e193).

tribution 4.0 International (CC BY 4.0): https://creativecommons.org/licenses/by/4.0/], which permits unrestricted use, distribution, and reproduction in

any medium, provided the original author and source are credited. The official and authorized publication credit sources, which will be duly enforced,

are as follows: official journal title Amphibian & Reptile Conservation; official journal website: amphibian-reptile-conservation.org.

Received: 1 October 2018; Accepted: 4 August 2019; Published: 8 November 2019.

Introduction

In Mexico, 132 species of plethodontid salamanders

have been recorded (AmphibiaWeb 2019), 42 of which

are Critically Endangered (CR) according to the Inter-

national Union for the Conservation of Nature (IUCN

2019). This conservation status may be assigned due to

various combinations of biological characteristics (e.g.,

restricted distribution or specific environmental require-

ments) and risk factors (e.g., habitat modification or

climate change; Stuart et al. 2008). In recent decades,

population declines have been observed or estimated for

several species (Frias-Alvarez et al. 2010) and in some

cases, despite intensive search efforts, finding them has

not been possible.

According to the IUCN (2019), 13 Critically Endan-

gered plethodontid species in Mexico have not been re-

ported in more than 10 years. Since 2010, six of these

species have been rediscovered: [sthmura naucampate-

pet! (Naturalista 2019), Chiropterotriton magnipes, C.

mosaueri, Pseudoeurycea ahuitzotl, P. tlahcuiloh, and

Thorius munificus (AmphibiaWeb 2019). However, so

far there have been no subsequent records of the remain-

ing seven species, including Thorius narismagnus. The

unknown status of these species highlights the need to

carry out sampling efforts in their historical localities

and to explore those areas with favorable environmental

conditions, in order to locate new populations (Sandoval-

Comte et al. 2012).

Thorius narismagnus (Shannon and Werler 1955)

is a minute salamander with a known distribution that

comprises only four localities, not more than 8 km apart,

on Volcan San Martin in the Los Tuxtlas region, Veracruz,

Mexico (Fig. 1), in an elevational range between 890

and 1,200 m asl (Hanken and Wake 1998). Thorius

narismagnus is considered as CR because the extent of

its occurrence is < 17 km? with reductions in the extent

and quality of its habitat, and due to a continuing decline

in the number of mature individuals (IUCN 2016).

The historical records of 7? narismagnus include 55

specimens collected between 1953 and 1976, and since

Correspondence. ! jal. herp@gmail.com, »-* paulinabanuelos@hotmail.es, *eduardo.pineda@inecol.mx;

+ sean.rovito@cinvestav.mx

Amphib. Reptile Conserv.

November 2019 | Volume 13 | Number 2 | e193

Aguilar-Lopez et al.

United States of America

a P|

gine og es

aay

Gulf of Mexico a

18°20'N 18°30'N 18°40'N

18°10'N

95°20'W 95°10'W

Mexico

Elevation (m)

[my 0-300

[i 301-600

fm 601-900

fy 901-1200

By 1201-1500

[>] 1501-1800

95°W 94°50'W 94°40'W

Fig. 1. Locations of historical collection localities and the new localities of Thorius narismagnus in Los Tuxtlas region, Mexico.

then this species has not been reported (IUCN 2016).

Rediscovery

As part of a study on the diversity and conservation

of amphibians in Veracruz, fieldwork was carried out

in a cloud forest in the community reserve Ejido Ruiz

Cortines, San Andrés Tuxtla, Veracruz (18°32'53"N,

95°09'16" W; 1,136 m asl) on Volcan San Martin (Fig. 1;

Fig. 2A) and in the private Ecological Reserve "La Otra

Opcion" Catemaco, Veracruz (18°22'32"N, 94°55'28"W;

1,075 m asl) on Volcan Santa Marta (Fig. 1; Fig. 2B).

At both sites searches for amphibians (08:00—12:00

and 20:00—00:00 h) were conducted in all terrestrial

microhabitats commonly used by these organisms

(Crump and Scott 1994).

In the Ejido Ruiz Cortines community reserve,

with a cumulative search effort of 72 person-hours in

September 2012, three individuals of the genus Thorius

were detected, two of which were collected (from which

a tissue sample was taken and the individuals were

subsequently preserved) and deposited in the Coleccion

de Anfibios y Reptiles del Instituto de Ecologia A.C.

(CARIE 0857, 1137; Fig. 2C). A sampling effort of 24

person-hours was carried out in July 2015 at this locality,

but there were no sightings of any minute salamanders.

The two collected specimens measured 17.6 and 11.4

Amphib. Reptile Conserv.

mm in SVL, the ratios between the length and width of

the nostrils were 1.14 and 1.09, with four and five free

intercostal grooves separating the adpressed fore and

hind limbs, respectively.

In the private ecological reserve La Otra Opci6on, with

cumulative search efforts of 96 person-hours in July 2015

and 66 person-hours in July 2017, three individuals of

Thorius were detected in well-preserved forests in each

year. The three individuals detected in 2015 were captured

(each one was measured and a sample of tail tissue was

taken) and subsequently released; these specimens had

SVL measurements of 15.4, 15.9, and 19.4 mm. The

three individuals found in 2017 were collected (CARIE

1251, 1258, 1259; Fig. 2D) and a sample of tissue was

taken from each; they measured 19.9, 22.5, and 21.2 mm

SVL, respectively. The proportions between the length

and width of the nostrils were 1, 1, and 1.15, respectively,

with 5.5 free intercostal grooves separating adpressed

limbs. Two females from La Otra Opcion (CARIE 1251

and 1259) had eight and five maxillary teeth, respectively,

while the adult male (CARIE 1258) lacked maxillary

teeth.

Identification of Specimens

The coloration in life of the specimens collected in the

Volcan San Martin and Volcan Santa Marta localities

November 2019 | Volume 13 | Number 2 | e193

Thorius narismagnus rediscovery in Mexico

Fig. 2. Habitat in the Volcan San Martin locality (A) and a specimen (C) collected from it in life (CARIE 0857). Habitat in the

—

. x, -

oe, ~ - . .

: _ * “he? -s - 1E os + & er .

Volcan Santa Marta locality (B) and a specimen (D) collected from it in life (CARIE 1251).

was light brown with a dark brown spike pattern in the

dorsum, dark brown color on the sides and a dark venter

with small white spots (Fig. 2C, 2D). With the exception

of the presence of maxillary teeth in two females, the

morphological characters and the coloration coincide

with the diagnosis proposed by Shannon and Werler

(1955) and Rovito et al. (2013) for Thorius narismagnus.

Although maxillary teeth are absent in most species of

Thorius, they are present in several species, including

T! smithi, which is relatively closely related to 7

narismagnus. Furthermore, at least two species (7.

grandis and T. omiltemi) have maxillary teeth present

only in females, and maxillary teeth are more common

in females of 7? minydemus than in males, which rarely

have them (Hanken and Wake 1998; Hanken et al. 1999).

Maxillary teeth were absent in a total of 18 specimens

from the type locality of 7) narismagnus on Volcan San

Martin as reported by Shannon and Werler (1955) and

Hanken and Wake (1998), suggesting that they do not

occur in either sex at that locality.

In order to confirm that both populations belong to

Thorius narismagnus, DNA was extracted from liver

tissue of one specimen from Volcan San Martin (the type

locality of TZ. narismagnus) and two specimens from

Volcan Santa Marta using a salt extraction protocol. A

Amphib. Reptile Conserv.

784 bp fragment of the cytochrome b gene was amplified

using primers MVZ15 and MVZ16 (Moritz et al. 1992).

PCR consisted of an initial denaturation step of 94 °C for

2 min, followed by 35 cycles of denaturation at 94 °C

for 30 sec, annealing at 48 °C for 1 min, and extension

at 72 °C for 1 min, with a final extension at 72 °C for 7

min. PCR products were purified using ExoSAP IT (USB

Corporation, Cleveland, Ohio, USA) and sequenced

using the BigDye v3.1 terminator cycle sequencing kit

(Applied Biosystems, Foster City, California, USA) on

an ABI 3730 capillary sequencer. Sequences were edited

using Geneious v8.1.8 (BioMatters, Auckland, New

Zealand), and sequences used in analysis were 750 bp

long after removing low-quality bases. Sequences for

other species of Thorius were obtained from GenBank

and sequences were aligned using Muscle v.3.8 (Edgar

2004). The pairwise GTR distance between sequences

from the two populations was calculated using PAUP

v4.165 (Swofford 2003). Sequences are deposited in

GenBank (Table 1).

The average pairwise divergence for cytochrome b

(cytb) between the two populations was 1.4% (Table

1). This level of divergence is comparable to, or lower

than, that seen between conspecific populations of

various species of Thorius. Several species of Thorius

November 2019 | Volume 13 | Number 2 | e193

Aguilar-Lopez et al.

Webi. 5

accession

mczai48745 | kcssaovs | 9.8 | 10.6] 12.0| 16.0| 73 | 160] 5.0 [19.5] 9.0 {1621 18.6] 13.7] 23.2| 19.0] 17.7] 17.7] 177] 12.5] 65 [103] 173] 59 | 68 |iss{ix0]i90] 66 | 51 fisstiz3| - | |

onus

Sonus

sicaudus

T. insperatus

T. maxillabrochus

T. minutissimus

T. lon

T. pennatulus

T. dubitus

T. lunaris

T. tlaxiacus

coed Mhoen!

Amphib. Reptile Conserv. 129 November 2019 | Volume 13 | Number 2 | e193

3 | T. pulmonaris

eae

Table 1. Percentage of average pairwise divergence for cytochrome b between the populations of Thorius species in GenBank.

Thorius narismagnus rediscovery in Mexico

from Oaxaca, including 7’ boreas, T. macdougalli, and

T. narisovalis, have substantially higher divergence

than that seen between the two populations from Los

Tuxtlas that were sequenced here. For example, two

populations of Thorius boreas separated by only 17 km

are 5% divergent for cytb (Rovito et al. 2013). The close

genetic similarity between the populations from Volcan

Santa Marta and from the type locality of Volcan San

Martin strongly suggests that these two populations are

conspecific.

Conservation Implications

The record from the Ejyido Ruiz Cortines community

reserve locality represents the rediscovery of T.

narismagnus, 36 years after the last reported record

(Hanken 1976: MVZ183028—-183035) at the type

locality on Volcan San Martin (Hanken and Wake 1998).

Additionally, the record from Volcan Santa Marta extends

the distribution range of this species 28 km southeast of

the closest known locality (Fig. 1). Because the specimens

reported here were found in primary vegetation, and

Diaz-Garcia et al. (2017) recorded 7. narismagnus

in mature forest but not in restoration areas and cattle

pasture in “La Otra Opcion” this species is probably not

able to survive in disturbed forest or even in moderately

disturbed forest. Of the other missing species that the

authors have found recently, only one (7) munificus)

was found in small, highly disturbed forest fragments

and within San Juan del Monte state reserve (Juarez-

Ramirez et al. 2016), while Chiropterotriton magnipes

and C. mosaueri were found in a cave in a national park

with only light to moderate habitat disturbance, and

Pseudoeurycea ahuitzotl and P. tlahcuiloh were found in

intact montane forest (AmphibiaWeb 2019).

To more fully understand the conservation status of

T. narismagnus, an exhaustive sampling effort through

time is needed to determine how the encounter rate of

this species varies throughout the year and whether it is

currently an uncommon species, and to obtain a more

accurate estimate of the population size. Extensive

fieldwork is also necessary in those areas with favorable

environmental conditions on both volcanoes to determine

the full distribution of this species. The extent of well-

preserved forest on Volcan San Martin is ~100 km?, while

on Volcan Santa Marta it is ~185 km? (INEGI 2016).

In addition, determining the presence of the chytrid

fungus Batrachochytrium dendrobatidis (Bd) and its

possible effect on the survival of these 7? narismagnus

populations is critical, because the presence of Bd in Los

Tuxtlas region has been confirmed (Mendoza-Almeralla

et al. 2015) and this pathogen is suspected of being linked

to the population declines of this species (IUCN 2016).

The presence of Bd has been reported in geographically

close species from Central Veracruz, such as Bolitoglossa

rufescens, Aquiloeurycea cephalica, and P. firscheini

(Van Roo et al. 2011), as well as P. nigromaculata

Amphib. Reptile Conserv.

and Thorius pennatulus, for which the presence of the

pathogen has been associated with declines in their

populations (Cheng et al. 2011).

Finally, the finding of two populations of a Critically

Endangered salamander species that has gone unrecorded

for almost four decades highlights the importance of

community and private reserves for harboring species in

imminent danger of extinction. Although both reserves

are relatively small (less than 200 ha) compared to

the Los Tuxtlas Biosphere Reserve in which they are

located, most of the land of these reserves is conserved

forest surrounded by modified environments. In that

sense, this work shows the complementarity between

the governmental and non-governmental reserves for the

protection of species at risk of extinction (see Garcia-

Bafiuelos et al. 2019), particularly those species with

restricted distributions and sensitivity to environmental

disturbances.

Acknowledgements——We are grateful to Ricardo

Luria, Luis Carrillo, Aristides Garcia, Juan Diaz, David

Gonzalez, and Rogelio Agapito for fieldwork support; and

to Ismael Guzman for initial molecular analysis. Wesley

Dattilo provided helpful suggestions that improved this

manuscript. Scientific collection permits were issued by

Secretaria del Medio Ambiente y Recursos Naturales

(SGPA/DGVS/03665/06 and SGPA/DGVS/03444/15).

Literature Cited

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Washington, DC, USA. 384 p.

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decline trends of Mexican amphibians. Biodiversity

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Thorius narismagnus rediscovery in Mexico

José L. Aguilar-L6pez was born in Mexico, and obtained his Bachelor’s degree at

Benemeérita Universidad Autonoma de Puebla (BUAP), and his M.Sc. degree at the

Instituto de Ecologia, A.C. (INECOL), both in Mexico. José recently obtained his Ph.D.

degree at INECOL, and he is interested in the diversity, ecology, and conservation of

amphibians and reptiles in tropical environments.

Paulina Garcia-Bafiuelos is a biologist, and recently obtained her Ph.D. degree at the

Instituto de Ecologia, A.C. (INECOL) in Xalapa, Veracruz, Mexico. Paulina obtained her

M.Sc. degree at the Instituto de Neuroetologia at the Universidad Veracruzana, the same

university where she obtained her Bachelor’s degree. She is interested in the current status

and conservation of the plethodontid salamanders of Mexico.

Eduardo Pineda obtained his Bachelor’s degree at the School of Sciences at the

Universidad Autonoma del Estado de México (UAEM) and his doctoral degree at the

Instituto de Ecologia, A.C. (INECOL), in Xalapa, Mexico. Eduardo’s research in the

INECOL is focused on understanding the relationship between the transformation of

tropical forests and biodiversity at different spatial scales, recognizing the importance of

conserved areas and modified habitats for maintaining the diversity of amphibians, and

assessing the current status through fieldwork of amphibian species that are in imminent

danger of extinction. Currently, Eduardo has several undergraduate and graduate students

addressing topics on the ecology and conservation of amphibians and reptiles in Mexico.

Sean Rovito is a professor at the National Laboratory of Genomics for Biodiversity

(Langebio, Cinvestav) in Irapuato, Mexico. Sean’s research focuses on diversification,

genomics, and conservation of Neotropical salamanders, particularly the plethodontid

salamanders of Mexico.

132 November 2019 | Volume 13 | Number 2 | e193

Amphibian & Reptile Conservation

13(2) [General Section]: 133-144 (e194).

Official journal website:

amphibian-reptile-conservation.org

urn:lsid:zoobank.org:pub:F6397C2F-00F 4-4885-810A-54D478A5A184

A new glassfrog (Centrolenidae: Hyalinobatrachium)

from the Topo River Basin, Amazonian slopes of the

Andes of Ecuador

14.* Juan M. Guayasamin, '*José Vieira, *Richard E. Glor, and *Carl R. Hutter

'Universidad San Francisco de Quito USFO, Colegio de Ciencias Biologicas y Ambientales COCIBA, Instituto BIOSFERA-USFO, Laboratorio de

Biologia Evolutiva, Campus Cumbaya, Casilla Postal 17—1200-841, Quito 170901, ECUADOR *Tropical Herping, Quito, ECUADOR *Department

of Ecology and Evolutionary Biology and Biodiversity Institute, University of Kansas, Lawrence, Kansas, USA *Department of Biology, University

of North Carolina, Chapel Hill, North Carolina, USA

Abstract.—A new species of glassfrog (Centrolenidae) is described from the San Jacinto River, an affluent

of the Topo River, on the Amazonian slopes of the Ecuadorian Andes. The new species, Hyalinobatrachium

adespinosai sp. nov., can be differentiated from all other centrolenids by the combination of its coloration

(transparent peritoneum and pericardium) and vocalization (call duration = 0.38-0.44 s, with 9-13 pulses per

call; dominant frequency = 4,645—5,001 Hz). However, H. adespinosai sp. nov. is morphologically cryptic with

H. anachoretus, H. esmeralda, and H. pellucidum, from which it differs by call traits (in H. anachoretus: call

duration = 0.32—0.37 s, with 5 or 6 pulses per call, dominant frequency = 4,670—4,800 Hz; in H. esmeralda: call

duration = 0.218—0.257 s, tonal call, dominant frequency = 4,739—5,580 Hz; in H. pellucidum: call duration = 0.112—

0.140 s, tonal, dominant frequency = 5,000—5,710 Hz). Biogeographically, the new species is separated from H.

anachoretus by a considerable distance and, also, the Maranon valley. Finally, following IUCN conservation

criteria, the status of the new species is considered as Data Deficient.

Keywords. Amphibia, Anura, Ecuador, Pastaza basin, phylogeny, Tungurahua Province

Resumen.—Describimos una nueva especie de rana de cristal (Centrolenidae) del rio San Jacinto, afluente del

rio Topo, en la vertiente amazonica de los Andes del Ecuador. La especie nueva, Hyalinobatrachium adespinosai

sp. nov., se diferencia de todos los centrolénidos por la combinacion de su coloracion ventral (peritoneo

y pericardio transparentes) y las caracteristicas de su canto (duracion del canto = 0.382-—0.430 s, con 9-13

pulsos por canto; frecuencia dominante = 4,645—5,001 Hz). Sin embargo, es morfologicamente criptica con H.

anachoretus, H. esmeralda y H. pellucidum, especies de las cuales difiere por su canto (en H. anachoretus:

duracion del canto = 0.32-—0.37 s, con 5 or 6 pulsos por canto, frecuencia dominante = 4,670—4,800 Hz; en H.

esmeralda: duracion del canto = 0.218-—0.257 s, tonal, frecuencia dominante = 4,739-—5,580 Hz; en H. pellucidum:

duracion del canto = 0.112-0.140 s, tonal, frecuencia dominante = 5,000—5,710 Hz). Finalmente, siguiendo los

criterios de la UICN, sugerimos que Hyalinobatrachium adespinosai sp. nov. sea ubicada en la categoria de

Datos Insuficientes.

Palabras clave. Amphibia, Anura, Cuenca del Pastaza, Ecuador, filogenia, Tungurahua Province

Citation: Guayasamin JM, Vieira J, Glor RE, Hutter CR. 2019. A new glassfrog (Centrolenidae: Hyalinobatrachium) from the Topo River Basin,

Amazonian slopes of the Andes of Ecuador. Amphibian & Reptile Conservation 13(2) [General Section]: 133-144 (e194).

tion 4.0 International (CC BY 4.0): https://creativecommons.org/licenses/by/4.0/], which permits unrestricted use, distribution, and reproduction in any

medium, provided the original author and source are credited. The official and authorized publication credit sources, which will be duly enforced, are

as follows: official journal title Amphibian & Reptile Conservation; official journal website: amphibian-reptile-conservation.org.

Received: 7 June 2019; Accepted: 12 October 2019; Published: 10 November 2019

Introduction traits. One of the most striking traits of this genus is

its complete ventral transparency, produced by having

The glassfrog genus Hyalinobatrachium (sensu Ruiz- a transparent ventral peritoneum (Ruiz-Carranza and

Carranza and Lynch 1991, as modified by Guayasamin = Lynch 1991; Cisneros-Heredia and McDiarmid, 2007;

et al. 2009) is one of the most charismatic anuran groups § Guayasamin et al. 2009). Males of all species in this

because of its peculiar morphological and behavioral genus also exhibit extended parental care towards

Correspondence. * jmguayasamin@usfq.edu.ec

Amphib. Reptile Conserv. 133 November 2019 | Volume 13 | Number 2 | e194

A new species of Hyalinobatrachium from Ecuador

fertilized egg clutches deposited on the leaves of trees

(see Delia et al. 2017). Extended male parental care, a

derived trait that has evolved at least twice in glassfrogs,

is coupled with egg deposition on the underside of leaves

in all Hyalinobatrachium and some Centrolene species

(Ruiz-Carranza and Lynch 1991; Guayasamin et al.

2009; Delia et al. 2017; Salgado and Guayasamin 2018).

Although assigning species to Hyalinobatrachium

is straightforward, distinguishing among members

of this genus is more complicated because species of

Hyalinobatrachium tend to be remarkably similar,

both morphologically and ecologically. In recent years,

species discovery in frogs has relied heavily on molecular

and acoustic traits (Castroviejo-Fisher et al. 2009, 2011;

Kubicki et al. 2015; Guayasamin et al. 2017). Calls, in

particular, are especially useful for distinguishing among

cryptic species since they function as efficient prezygotic

mating recognition signals (Narins and Capranica 1976;

Duellman and Trueb 1994; Zakon and Wilczynski 1988:

Wilczynski and Ryan 1999; Wells 2007). As previously

demonstrated in glassfrogs, the acoustic differences

between species are often more pronounced than

morphological differences (Escalona-Sulbaran et al.

2018).

We describe a new species of Hyalinobatrachium that

is morphologically cryptic with H. anachoretus Twomey,

Delia, and Castroviejo-Fisher 2014, H. esmeralda Ruiz-

Carranza and Lynch 1998, and H. pellucidum (Lynch and

Duellman 1973). The new species, which is known from

a single locality in the Topo River basin, is differentiated

from these two species by its call and genetics.

Materials and Methods

Ethics statement. Research was conducted under

permits NoMAE-DNB-CM-2015-2017, 019-2018-IC-

FAU-DNB/MAE, and 018-2017-IC-FAU-DNB/MAE,

issued by the Ministerio del Ambiente del Ecuador. The

study was carried out in accordance with the guidelines

for use of live amphibians and reptiles in field research

(Beaupre et al. 2004), compiled by the American

Society of Ichthyologists and Herpetologists (ASIH),

the Herpetologists’ League (HL), and the Society for the

Study of Amphibians and Reptiles (SSAR).

Taxonomy and species concept. Glassfrog generic

and family names follow the taxonomy proposed by

Guayasamin et al. (2009). Species are considered

as segments of separately evolving metapopulation

lineages, following the conceptual framework developed

by Simpson (1951, 1961), Wiley (1978), and de Queiroz

(2007). Assessing whether a given population is an

independent lineage is a non-trivial task, especially

when working with closely related taxa. In such cases,

analyzing many different sets of characters provides

tools for supporting species hypotheses (Dayrat 2005; de

Queiroz 2007; Padial et al. 2009).

Morphological data. Lynch and Duellman (1973) and

Cisneros-Heredia and McDiarmid (2007) are followed

for the diagnosis and description of the new species. The

webbing formula follows Savage and Heyer (1967), as

Amphib. Reptile Conserv.

modified by Guayasamin et al. (2006). The taxonomy of

centrolenid frogs follows the proposal by Guayasamin

et al. (2009). Comparisons were made between various

Hyalinobatrachium specimens (see Appendix 1)

housed at the following collections: Instituto de Ciencia

Naturales, Universidad Nacional de Colombia, Bogota,

Colombia (ICN); University of Kansas, Museum of

Natural History, Division of Herpetology, Lawrence,

Kansas, USA (KU); Museo de Zoologia, Universidad

Tecnologica Indoamérica, Quito, Ecuador (MZUTI);

National Museum of Natural History, Smithsonian

Institution, Washington, DC, USA (USNM):; and Museo

de Zoologia, Universidad San Francisco de Quito, Quito,

Ecuador (ZSFQ). Morphological measurements were

taken with a Mitutoyo® digital caliper to the nearest

0.1 mm, as described by Guayasamin and Bonaccorso

(2004), except when noted, and are as follows: (1)

snout—vent length (SVL); (2) tibia length; (3) foot

length; (4) head length; (5) head width; (6) interorbital

distance (IOD); (7) upper eyelid width; (8) internarial

distance; (9) eye-to-snout distance; (10) eye diameter;

(11) tympanum diameter; (12) radioulna length; (13)

hand length; (14) Finger I length; (15) Finger II length

= distance from outer margin of palmar tubercle to tip

of Finger II; and (16) width of disc of Finger III. Sexual

maturity was determined by the presence of vocal slits

and calling activity in males.

Bioacoustics. Sound recordings were made with an

Olympus LS-10 Linear PCM Field Recorder and a

Sennheiser K6—ME 66 unidirectional microphone. The

calls were recorded in WAV format with a sampling rate

of 44.1 kHz/s with 16 bits/sample. Measurements and

definitions of acoustic variables follow Kohler et al.

(2017). Notes were divided into two classes—“pulsed”

and “tonal”—based upon the distinct waveforms on the

oscillogram (see Hutter and Guayasamin 2012). Pulsed

(also termed peaked) notes are defined as those with one

or more clear amplitude peaks and amplitude modulation

(i.e., visible increases and decreases in amplitude on the

oscillogram throughout the call). In contrast, tonal notes

are defined as those with no clear amplitude peak (Dautel

et al. 2011). In this study the call of Hyalinobatrachium

pellucidum (Lynch and Duellman 1973) is also described

from an individual (USNM 286708) recorded at the type

locality of the species (Rio Azuela, 0.1167°S, 77.617°W,

1,740 m, Napo province, Ecuador) by Roy McDiarmid;

the recording is deposited at the Cornell University

Macaulay Library (ML Catalogue No. 202401).

Evolutionary relationships. Mitochondrial sequences

(16S) were generated for four individuals (ZSFQ 1647—

48, 1650-51) of the new species of Hyalinobatrachium.

Extraction, amplification, and sequencing protocols were

as described in Guayasamin et al. (2008). The sequences

obtained were compared with all those available for other

species of glassfrogs (family Centrolenidae) and its sister

taxon Allophrynidae (see Austin et al. 2002; Guayasamin

et al. 2018). These sequences were downloaded from

GenBank (https://www.ncbi.nlm.nih.gov/genbank/), and

were generated mostly by Guayasamin et al. (2008),

Castroviejo-Fisher et al. (2014), and Twomey et al.

November 2019 | Volume 13 | Number 2 | e194

Guayasamin et al.

(2014), but also included data from the newly described

H. yaku Guayasamin et al. 2017 and H. muiraquitan de

Oliveira and Hernandez-Ruz 2017. Sequences were

aligned using MAFFT v.7 (Multiple Alignment Program

for Amino Acid or Nucleotide Sequences: http://mafft.

cbre.jp/alignment/software/), with the Q-INS-1 strategy.

MacClade 4.07 (Maddison and Maddison 2005) was

used to visualize the alignment (no modifications were

necessary). Maximum likelihood (ML) was run in the

IQ-TREE 1.5.5 software (Nguyen et al. 2015). The best-

fitting nucleotide substitution model was implemented

using ModelFinder within IQ- TREE (Kalyaanamoorthy et

al. 2017), which groups partitions with the same model

and similar rates, and simultaneously searches the model

and tree space. Node support was assessed via 1,000 ultra-

fast bootstrap replicates, a method that shows less bias

than other support estimates (Minh et al. 2013). Ultra-fast

bootstrapping also leads to straightforward interpretation

of the support values (e.g., support of > 95 bootstrap

should be interpreted as significant; Minh et al. 2013).

Results

Phylogenetic relationships. Based on the Bayesian

Information Criterion, the best-fit model for this dataset

was GTR+F+R5. Rate parameters were estimated as

follows: A—C: 2.690, A-—G: 9.067, A-T: 3.078, C-—G:

0.346, C—T: 23.835, and G—T: 1.000. Base frequencies

were: A = 0.346, C = 0.239, G=0.181, and T = 0.234.

The phylogeny (Fig. 1) confirms the placement of

the new species within the genus Hyalinobatrachium.

The new species, described below, is inferred as part of

a clade formed by four species that have very similar

morphologies: the new species (described below) + H.

anachoretus + H. pellucidum + H. yaku.

Species description

Hyalinobatrachium adespinosai new species

urn:lsid:zoobank.org:act:46C6CBB4-ECE3-470E-A 15C-4BB36A3301DE

Suggested English name: Adela’s Glassfrog

Suggested Spanish name: Rana de Cristal de Adela

Holotype. ZSFQ 1648 (JMG 583, Fig. 2), adult male

from riverine vegetation along the San Jacinto River

(1.3447°S, 78.1814°W; 1,795 m asl), Tungurahua

Province, Ecuador, collected by CRH, REG, and KC on

4 August 2017.

Paratypes. ZSFQ 1650-52, 1647, adult males with same

data as holotype.

Generic placement. The new species is placed in the

genus Hyalinobatrachium (Ruiz-Carranza and Lynch,

1991, as modified by Guayasamin et al. 2009) on the

basis of morphological and molecular data. The main

diagnostic phenotypic traits of Hyalinobatrachium are:

(1) ventral parietal peritoneum completely transparent;

(2) digestive tract and bulbous liver covered by

iridophores; (3) humeral spines absent; (4) white bones

Amphib. Reptile Conserv.

in life; (5) males call from the underside of leaves;

(6) females place the eggs on the underside of leaves;

and (7) males provide extended parental care. All the

aforementioned characteristics are shared by the new

species. Additionally, analyses of the mitochondrial

16S gene place the new species as a close relative of

other Hyalinobatrachium species (Fig. 1); thus, generic

placement in Hyalinobatrachium is unambiguous.

Diagnosis. Within the genus HAyalinobatrachium,

the new species is diagnosable mainly by having a

transparent pericardium. However, the new species

is morphologically cryptic with three closely related

taxa (H. anachoretus, H. pellucidum, H. esmeralda).

Based on comparisons with specimens examined (see

Appendix 1), all these species display a similar size and

color pattern (pale green dorsum with yellow dots and

a transparent venter and pericardium; red heart visible

ventrally). However, calls between species diverge

noticeably; the major difference is the structure of the

call, with two species (H. adespinosai sp. nov. and H.

anachoretus) having pulsed calls and the others having

tonal vocalizations (Fig. 3; Table 1). The call of #7.

adespinosai sp. nov. is further differentiated from that

of H. anachoretus by being longer, having more pulses

per note, and being produced at a higher rate (Table

1). Toe webbing (Toe IV) is less extensive in the new

species (2'3 IV 2°) than in H. anachoretus (1* IV 1°;

Twomey et al. 2014). Additionally, the new species and

H. anachoretus are separated by considerable distance

(airline distance = 473 km), including one of the most

important biogeographic barriers in South America, the

Marafion valley (see Duellman 1999; Winger and Bater

2015 and references therein). Uncorrected p genetic

distances for the mitochondrial gene 16S between

H. adespinosai sp. nov. and its closest relatives are

summarized in Table 2.

Characterization. The following combination of

characters is found in Hyalinobatrachium adespinosai

sp. nov.: (1) dentigerous process of the vomer lacking

teeth; (2) snout truncate in dorsal and lateral views:

(3) tympanum barely visible, hidden under skin, with

coloration similar to that of surrounding skin; (4)

dorsal skin shagreen; (5) ventral skin areolate; cloacal

ornamentation absent, paired round tubercles below vent

absent; (6) parietal peritoneum transparent; pericardium

with thin layer of iridophores (in life, a red heart is mostly

visible ventrally); liver, viscera, and testes covered

by iridophores; (7) liver bulbous; (8) humeral spines

absent; (9) hand webbing formula: I (2-3) — (2—2*) IT

(1-1*) —3'° III (2-2*) — (2—2) IV; (10) foot webbing

moderate; webbing formula: I 1— (17°-2-) I (1-1-) —

(2—2"3) IIT (1-1*) —(2*-2!%) IV 2*— (1*-1"%) V; (11)

fingers and toes with thin lateral fringes; ulnar and tarsal

folds present, but low and difficult to distinguish, with

thin layer of iridophores that extends to ventrolateral

edges of Finger IV and Toe V; (12) nuptial excrescence

present as a small pad on Finger I (Type V), prepollex

not enlarged; prepollical spine not projecting (spine not

exposed); (13) when appressed, Finger I longer than II;

(14) diameter of eye about two times wider than disc on

November 2019 | Volume 13 | Number 2 | e194

A new species of Hyalinobatrachium from Ecuador

Celsiella revocata (EU663019)

Celsiella vozmedianoi (EU663025)

H. cappellei (EU663040)

H. iaspidiense (EU663047)

H. tricolor (EU663027)

H. taylori (EU663056)

H. mondolfii_ (EU663050)

H. munozorum (EU663034)

H. pallidum (€U663052)

H. carlesvilai (KM068270)

H. carlesvilai (KM068271)

H. carlesvilai (EU663030)

H. fleischmanni (DQ283453)

H. fleischmanni (Jx564869)

H. fleischmanni (EU663045)

H. fleischmanni (EU663044)

H. tatayoi (EU663055)

H. muiraquitan (KY310571)

H. muiraquitan (KY310570)

H. kawense (EU663029)

H. duranti (EU663041)

H, ibama (EU663048)

Hyalinobatrachium sp (EU447290)

H. orientale (EU447289)

H. guairarepanense (KF534363)

H. orocostale (EU447284)

H. fragile (€U447286)

H. chirripoi (KF604294)

H. chirripoi (EU663037)

H. chirripoi (EU663038)

H. aff. colymbiphylium (kM068297)

H., aff. colymbiphyllum (KF604300)

H. aff. colymbiphyllum (FJ784471)

H. colymbiphyllum (FJ784346)

H. colymbiphyllum (FJ784475)

H. colymbiphyllum (FJ784527)

H. colymbiphyllum (EU663039)

H. anachoretus (KM068300)

H. anachoretus (KM068268)

H. adespinosai sp. nov. (ZSFQ-1648: MN604036)

H. adespinosai sp. nov. (ZSFQ-1651: MN604037)

H. adespinosai sp. nov. (ZSFQ-1647: MN604038)

H. adespinosai sp. nov. (ZSFQ-1650: MN604039)

H. pellucidum (GQ142065)

H. pellucidum (KM068252)

H, yaku (MF002066)

: H. yaku (MF002067)

H. aff. esmeralda (EU663036)

H. aff. esmeralda (KP149361)

H. bergeri (EU663033)

——<$$<———. Hyalinobatrachium sp (KM068298)

100

———— Hyalinobatrachium sp (KM068299)

H. vireovittatum (KF604303)

H. talamancae (EU663054)

0.02 substitutions/site

H. aff. bergeri (EU663026)

H. aureoguttatum (EU663025)

H. valerioi (EU663058)

Fig. 1. Phylogenetic relationships of Hyalinobatrachium inferred from the 16S mitochondrial gene under ML criteria. All sequences

were downloaded from GenBank, except for those of the new species. GenBank codes are listed next to each terminal. Associated

locality data is available at GenBank, as well as in Guayasamin et al. (2008), Castroviejo-Fisher et al. (2014), and Twomey et al.

(2014),

Finger III; (15) coloration in life: dorsal surfaces pale

yellowish green with small pale yellow spots and minute

gray to black melanophores; bones white; (16) coloration

in preservative: dorsal surfaces pale cream with minute

melanophores; (17) iris coloration in life: white with pale

yellow hue and numerous minute lavender spots; (18)

melanophores absent from most fingers and toes, but

present on Finger IV and Toes IV and V; (19) males call

Amphib. Reptile Conserv.

from underside of leaves; advertisement call consisting

of single note, distinctly pulsed (9-13 pulses per call),

with duration of 0.382—0.430 s, and dominant frequency

at 4,645-5,001 Hz; (20) males attend egg clutches

located on the underside of leaves overhanging streams;

clutch size of 22 embryos (n = 1); (21) SVL in adult

males 20.5—22.2 mm (n = 3), females unknown; and (22)

enameled tubercles absent from sides of head.

November 2019 | Volume 13 | Number 2 | e194

Guayasamin et al.

Fig. 2. Hyalinobatrachium adespinosai sp. nov. in life,

holotype.

Description of the holotype. ZSFQ 1648, adult male

with SVL 22.2 mm. Head wider than long (head width

38% of SVL; head length 77% of head width). Snout

truncate in dorsal and lateral views. Loreal region

flat and nearly vertical, nostrils slightly protuberant,

elliptical; internarial region concave anterodorsally;

canthus rostralis well defined. Eyes small, directed

anterolaterally, eyes about 45° relative to midline (where

Amphib. Reptile Conserv.

Amplitude (kU)

Frequency (kHz)

1:03.4 1:03.6 1:03.8

Time (m:s)

Fig. 3. Call of Hyalinobatrachium adespinosai sp. nov.,

holotype, recorded in field conditions at the type locality. Air

temperature: 18 °C.

anterior-facing eyes would be 90° relative to midline).

Tympanum annulus barely visible through the skin;

tympanic membrane differentiated and pigmented as

surrounding skin. Dentigerous processes on vomers

absent; choanae large, oval, separated widely (distance

about the same as between nostrils); tongue round, white

in preservative, anterior 4/5 attached to mouth; vocal slits

present, extending along floor of mouth lateral to tongue;

enameled glands absent from lower part of upper jaw.

Ulnar fold present, with a thin layer of iridophores, and

continuing along the external edge of Finger IV; humeral

spine absent. Relative lengths of fingers: I < II < IV <

II; finger discs rounded, about the same size as discs on

toes, disc on Finger II 54% of eye width; finger webbing

reduced between Fingers I-III, moderate between

Fingers III and IV, with formula I 3—2* II 1*—3'° I]

2 — 2 IV. Prepollex concealed; subarticular tubercles

round, faint; few small supernumerary tubercles present,

palmar tubercle round and small, thenar tubercle ovoid;

nuptial excrescences present as a small pad on external

edge of Finger I (Type V). Hind limbs slender, tibia

length 58% of SVL; tarsal fold present, with thin layer of

iridophores; discs of toes round, inner metatarsal tubercle

small; outer metatarsal tubercle round, but very difficult

to distinguish. Webbing formula of feet: I 1— 2> II 1—2'"

I 1’—2'71V—2*— 1V. In preservative, dorsal skin

peppered with small dark lavender melanophores; dorsal

skin shagreen; skin on venter areolate; cloacal opening

at level of upper thighs, cloacal ornamentation present

as a lightly enameled cloacal fold and small tubercles.

Parietal peritoneum transparent; pericardium with a very

thin layer of iridophores that, in life, exposes a red heart:

urinary bladder lacking iridophores; liver, viscera, and

testes fully covered by iridophores. Kidneys rounded,

approximately bean-shaped; liver bulbous.

Coloration in life. Dorsal surfaces apple green to

yellowish green with diffuse yellow spots and minute

gray to black melanophores. Melanophores absent

from fingers and toes, except Finger IV and Toes IV

and V. Ventrally, parietal peritoneum and pericardium

transparent, with a red heart always visible, even

November 2019 | Volume 13 | Number 2 | e194

A new species of Hyalinobatrachium from Ecuador

when a very thin layer of iridophores is present on the

aS 2 = : : Erne I :

3 5 % g pericardium of some individuals. Visceral peritoneum

S < E S at ic 2 fi C of gall bladder and urinary bladder transparent; hepatic

S} ae |e & a. and visceral peritonea white; ventral vein red. Iris pale

Sm Sa a a = ellowish white, with numerous minute lavender spots.

uae, y ;

om Bones white.

So] 8 bs Dies

~ clELR 3 Of SO o eae 3 Op . : :

wo less a 4-8 2 PAS 28 Coloration in preservative. Dorsal surfaces cream

& Els oS a= a= > > H o : : i

2 SEES E ‘ap Fa = S90 8 dotted with minute dark lavender melanophores; venter

a8 | 8 is oS ae ob ar~ A § ; ; i ee

oe erg = Fw uniform cream; visceral peritoneum lacking iridophores;

x dD & - ee 2 ; i ; cme Nig

0 © pericardium with a very thin layer of iridophores. Iris

£e = T em T T silvery white with minute lavender melanophores.

BS a

acol]ees Cor) 2 =o ws wes

S$s|223. |82 22 a eat M3

—~S)EeePe PAH + "i ao oH Measurements. Measurements of the type series are

oSijegee valley S at wy H vay ; :

Sania ee 0 & S 2s 38 d & shown in Table 3.

Sr |fie [as + oe 6S BAe

23 - < i el % Le es Ete red Ae

3 5 Variation. Variation in hand webbing is as follows: I

nN —| aj

26 Zs Poe Bee (2-3) — (2-2") LICL IR) —3"3 TI] (2-2") — (2-2) IV.

vQ|35 Z Steg dente Lie Foot webbing variation is as follows: I 1— (17°-2-) II

SY a n = =

SS \E2 ae RE (1-1-) —(2—2!) IIT (1-1*) —(2*-2!) IV 2*— (1*-1'*)

gs V.

for}

ae o™~ o™~

a2] = S a = lows ya

zselea On - = ae am Vocalizations (Fig. 3). The description is based on

ao Oo é = wy ia = on oH oo 4H H = : ih

ee ee T 2% A 7S silts. it. recordings from nine individuals (Codes LBE-C: 048—

pate | ae = Ga Pe say 0 he call of Hyalinobatrachium adespinosai

o/s ole a uv WU 57). The call of Hyalinobatrachium adespinosai sp.

al ae we cs re] by nov. has a striking resemblance to the chirp of a cricket,

See and was often confused for one in the field. Each call is

8/3 +H = ¥ = composed of a single and high-pitched pulsed note, and

on he Oo : Se ‘

3 = 12 mee . 4 aus + has a duration of 0.38-0.44 s (xX = 0.38 + 0.017). Time

n = so N | oe Oe . —_

as lg ajo |S D TIS f between calls varied from 2.0-11.0 s (X = 4.58 + 2.3).

G = & — ae ef The fundamental frequency, the same as the dominant

Seni, frequency, is at 4,645—5,203 Hz (X = 4,855 + 152). There

= | , (is

Sal’ is no frequency modulation. The first harmonic is at

Ow S : *

24 = 3 3 = = = 9,336—9,754 Hz and the second harmonic is at 14,159-—

a n nA fon i= S

Rep le = = e e 2 14.444 Hz

_ Oo @ & & am ome cH > ;

Fels

s = Ecology. All individuals of the new species were found

S 5/5 on the underside of leaves of riverine vegetation along the

gS eR 2. " ' * ' :

Ss e/8 3 x - ¥. = = San Jacinto River. The section of river was fast-flowing

oe) =) ae ‘ : A

S&|5 and had visible rapids. Although the population is locally

Ss + 7

8 8 abundant (as heard from numerous advertisement calls

eS Sack .

ron le oe eats a mH es uh individuals are very difficult to observe because they are

SS(EPFESISRS FS SRF S8S Sag lly found at th level (4-16 mab d

Aes es 2.8 Siok S SAY cst cs8a usually found at the canopy level ( m above groun

oS /Sts ely Cs a eres lcs ye ; . :

5 §/seess|S Le & Rupe eS NO? level). The type series consists of males exclusively; they

a. RS Crm vi a a were calling in the months of July and August. One male

3% <a (ZSFQ 1648) was apparently guarding an egg clutch

a containing 22 embryos; both the adult male and the egg

Csletss IE Si = 2 =

SBlEsts |& = = = S clutch were on the same leaf most of the time, but the

Ooh ler ay ‘ male also moved to nearby leaves (Fig. 4).

o.5 A

aes

ae) eae :

Ss 3 g 3 « : 30 os g& Distribution. Hyalinobatrachium adespinosai_ sp.

Ss a 4 6 . ‘ :

ane le § Ex sao sg, BEI § nov. is only known from the type locality: San Jacinto

[a0] — N a) . A i) :

he = eas soc E 88s ee%, FS River (1.3447°S, 78.1814°W; 1,795 m asl), Tungurahua

+2 2 Ge | 2s San Ct en : ‘

on be Ros “So seat BES kg Province, Ecuador (Fig. 5).

& 16 gee poo, Cree SN Ss 3§

ay eet eee EES ASE

== ae i . “es AS Evolutionary relationships. The phylogenetic

28 analyses recover Hyalinobatrachium adespinosai sp.

= = ¢. nov. haplotypes as sister to haplotypes sampled from

pan ae i in

5 8 5 s . S H. anachoretus and nested within other members of

> S S Ss = ;

= 8 & s S iS a monophyletic clade comprised of all other sampled

n 7.) : s : F - :

2 s z 8. S 5 3 $ species of Hyalinobatrachium (Fig. 1). The most closely

iS 3 & a ~ ~ = ~ related species to H. adespinosai sp. nov. share several

Amphib. Reptile Conserv. 138 November 2019 | Volume 13 | Number 2 | e194

Guayasamin et al.

\ f

~~ .

- >

< \ >

ag ay

egg clutch; other males were observed on the same leaf as the egg clutch. (B) Close-up of the egg clutch. (C) Spider predation on

an unattended egg clutch.

morphological traits, including a red heart exposed

ventrally (H. adespinosai + H. anachoretus + H.

pellucidum + H. yaku).

Etymology. The specific epithet adespinosai honors

Adela Espinosa, an Ecuadorian conservationist and

board member of the Jocotoco Foundation (http://

www.jocotoco.org). Adela’s work has focused on the

conservation of species and ecosystems. The new

glassfrog described here is found only within the limits

of a natural reserve owned by Adela and her husband,

Antonio Paez. We are delighted to recognize Adela’s

devotion to nature with this marvelous species.

Conservation status. Available information is

insufficient to fully assess the conservation status of

Hyalinobatrachium adespinosai sp. nov. Therefore,

following IUCN criteria, this species is considered as

Data Deficient. The herpetological museums that house

specimens collected near the type locality (Topo basin)

were consulted, but there were no additional specimens

of the new species. Although this might suggest a

conservation category other than Data Deficient, we

actually prefer to maintain this status because the new

Species 1s very difficult to find (1.e., a canopy specialist).

Therefore, in this case, absence in nearby localities

where herpetological surveys have been carried out does

not necessarily indicate a true absence of the species.

Discussion

Morphological stasis is expected in species under similar

ecological conditions, whereas traits associated with

social signaling tend to evolve more rapidly (Winger

and Bater 2015; Arnegard et al. 2010; Safran et al.

2013; Escalona-Sulbaran et al. 2018). Species in the

glassfrog genus Hyalinobatrachium exhibit a striking

morphological homogeneity (see Ruiz-Carranza and

Table 2. Genetic distances (uncorrected p matrix for 16S, 813 base pairs) between Hyalinobatrachium adespinosai sp. nov. and

closely related species.

H. adespinosai H. anachoretus

H. adespinosai 0.000-0.001

HZ. anachoretus 0.010-0.011 0.000

H.. esmeralda 0.0254—0.0272 0.0272

H. pellucidum 0.032-0.034 0.029-0.037

HA. yaku 0.036—0.037 0.033—0.041

Amphib. Reptile Conserv. 139

H. esmeralda H. pellucidum HA. yaku

0.000

0.034—0.040 0.007—0.009

0.038—0.040 0.025—0.030 0.000—0.001

November 2019 | Volume 13 | Number 2 | e194

A new species of Hyalinobatrachium from Ecuador

ok ft A -76°

Colombia

Fig. 5. Distribution of Hyalinobatrachium adespinosai sp. nov. in Ecuador.

Lynch 1998; Guayasamin et al. 2009; Castroviejo-Fisher

et al. 2011), perhaps because of the constraints associated

not only with their similar ecology, but also with their

derived reproductive strategy (prolonged parental care

on the underside of leaves). The obvious consequence is

that traditional morphological trait-based criteria provide

an underestimation of the true biological diversity of the

genus. In contrast, call traits in centrolenids have shown

more variation that morphology (Escalona-Sulbaran et

al. 2018). Acoustic signals can diverge because of the

effects of multiple mechanisms, including drift (e.,

isolation-by-distance), natural selection (1.e., adaptation

to local ecological conditions, reinforcement, character

displacement), and/or sexual selection (1.e., sensory

exploitation, divergent female choice; reviewed by

Wilczynski and Ryan 1999; Wells 2007; Prum 2017;

Kohler et al. 2017). However, this study represents

another example of how vocalizations can be extremely

useful for species discovery.

Given the lack of information for Hyalinobatrachium

adespinosai sp. nov., we consider the species as Data

Deficient, following the IUCN criteria. The species is

Amphib. Reptile Conserv.

locally abundant at the type locality and as currently

known has a restricted distribution. However, given that

the species is usually found at the canopy level, it is

extremely difficult to locate individuals of this species,

So we cannot infer its true distribution based solely on the

lack of prior collection.

Establishing clear biogeographic patterns in

groups where new species are often being described

is challenging. However, Hyalinobatrachium species

are generally found in the lowlands while Centrolene

and Nymphargus species are predominantly Andean

(Guayasamin et al. 2009; Hutter et al. 2017). In contrast

to these general patterns, the clade formed by H.

adespinosai sp. nov. + H. anachoretus + H. pellucidum

+ H. yaku + H. esmeralda is mostly found on the

Amazonian slopes of the Andes (except H. yaku). Since

tropical species tend to have narrow thermal niches (Shah

et al. 2017, Polato et al. 2018), the linearity of the Andean

mountain range might promote speciation by reducing

contact and gene flow among parapatric populations (see

Fig. 6), as suggested by Graves (1988). Similar patterns

(i.e., closely related species along the same slope of

November 2019 | Volume 13 | Number 2 | e194

Guayasamin et al.

Lineary of the Andes:

Loss of connectivity because of random local extinction

Lineary of the Andes:

Loss of connectivity because of valleys (ecological barriers)

3 Species

Species C

eee SSS

Fig. 6. Schematic graph illustrating how the linearity of the Andes facilitates the speciation process.

the Andes) have also been observed in other glassfrogs

(e.g., Nymphargus; Guayasamin et al. 2019) and birds

(Bonaccorso 2009; Benham et al. 2015; Cadena et al.

Table 3. Meristic variation of Hyalinobatrachium adespinosai

sp. nov. (in mm).

2019). ZSFQ ZSFQ ZSFQ

1647 1651 1647

Acknowledgments.—Two reviewers (Alejandro Arteaga (holotype)

and an anonymous reviewer) provided comments that ae Male Male Rigie

greatly improved this article. Research permits were

issued by the Ministerio de Ambiente del Ecuador SVL 22.2 20.7 20.5

(MAE-DNB-CM-2015-0017, 019-2018-IC-FAU-DNB/ Femur 12.8 12.0 12.1

USNM for providing access to specimens housed at their

collections. This study was supported by the Universidad Foot m0 sue 10P

San Francisco de Quito (Equipamiento del Laboratorio Head length 6.5 6.4 6.2

de Biologia Evolutiva, project ID 5467; Ranas de Cristal: Head width 84 rg PO

Taxonomia, Evolucion y Conservacion, project ID 5466), IOD 28 25 6

and the Programa Inédita from Secretaria de Educacion

Superior, Ciencia, Tecnologia e Innovacion (Project: Upper eyelid 1.6 1.4 1.6

Respuestas a la crisis de biodiversidad: la descripcion Internarinal 1.8 1.7 1.7

de especies como herramienta de conservacion). Finally, distance

our most warm thanks to Walt and Linda Jennings for Eye diameter 16 23 23

supporting the wonderful conservation efforts in Ecuador. EeePONt 39 28 3.0

, . distance

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November 2019 | Volume 13 | Number 2 | e194

A new species of Hyalinobatrachium from Ecuador

Appendix 1. Examined Specimens

Hyalinobatrachium esmeralda: Colombia: Boyaca Department: Municipio de Pajarito, Inspeccién Policia Corinto,

finca'El Descanso’, quebrada'La Limonita', 1,600—1,650 m, ICN 9592-94, 9596, 9602-03 (type series of H. esmeralda).

Hyalinobatrachium pellucidum: Ecuador: Morona Santiago Province: Nueva Alianza, Finca Santa Catalina

(78.1335°W, 2.100°S; 1,305 m), Limite del Parque Nacional Sangay, MEPN 14706. Quebrada del Rio Napinaza

(78.4070°W, 2.9266°S, 1,100 m), QCAZ 42000; km 6.6 on the Limon-Macas road (ca. 2.92816°S, 78.344°W; 1,013

m), QCAZ 29438; 6 km N of Limon, QCAZ 25950. Sucumbios Province: Rio Azuela (0.1167°S, 77.6167°W, 1,740

m), Quito-Lago Agrio road; KU 164691 (holotype), USNM 286708—10; Rio Reventador, USNM 286711-—12. Zamora

Chinchipe Province: Cordillera del Condor, Miazi Alto (4.25044°S, 78.61356°W; 1,282 m), QCAZ 41560-61, 41648.

Hyalinobatrachium munozorum: Ecuador: Sucumbios Province: Santa Cecilia (00°03'N, 76°58'W; 340 m), KU 118054

(holotype), 105251, 123225, 150620 (paratypes), 152488—-89, 155493-96, 175504. Orellana Province: Tiputini

Biodiversity Station, ZSFQ DFCH-USFQ D105. Colombia: Meta Department: Meta, ICN 5031-34, 39503. Amazonas

Department: Leticia, ICN (field number JMR 4119).

Hyalinobatrachium yaku: Ecuador: Pastaza Province: stream affluent of the Kallana river (1.4696°S, 77.2784°W;

325 m), MZUTI 5001 (holotype), 5002 (paratype). Orellana Province: Timburi-Cocha Research Station (0.4800°S,

77.2829°W; 300 m) near San José de Payamino, QCAZ 55628, QCAZ 53352, 53354, 56664. Napo Province: Ahuano

(1.0632°S, 77.5265°W; 360 m), ZSFQ 02322.

Juan M. Guayasamin is a professor at Universidad San Francisco de Quito, Ecuador, and

codirector of the Laboratory of Evolutionary Biology. Juan obtained his Master’s and Ph.D.

degrees in ecology and evolutionary biology from the University of Kansas (Lawrence,

Kansas, USA) under the supervision of Dr. Linda Trueb. He is member of the Ecuadorian

Academy of Sciences and has published more than 80 scientific papers on evolution,

systematics, biogeography, and conservation of Neotropical animals, mainly amphibians.

Jose Vieira is a field biologist, wildlife photographer, and tour leader from Venezuela. From

a young age, Jose became passionate about nature, particularly amphibians and reptiles. This

passion led him to participate in countless field expeditions in his native country, and from

them Jose has contributed many herpetological specimens to the Museo de Historia Natural

La Salle. Currently, his contributions to science continue in Ecuador with the rediscovery of

the critically endangered Ate/opus bomolochos and A. nepiozomus, and his expeditions to

remote areas of the country to work on various herpetological projects of Tropical Herping

and Universidad San Francisco de Quito.

Richard E. Glor is a Professor in the Department of Ecology and Evolutionary Biology,

and a Curator in the Biodiversity Institute, at the University of Kansas (Lawrence, Kansas,

USA). Richard studies the evolution of species diversity, primarily through work on West

Indian anole lizards. He received his Bachelor’s degree from Cornell University (Ithaca,

New York, USA), his doctorate in Ecology and Evolutionary Biology from Washington

University (St. Louis, Missouri, USA), and conducted postdoctoral research in the Center

for Population Biology at University of California, Davis.

Carl R. Hutter recently obtained his Ph.D. from the University of Kansas (Lawrence,

Kansas, USA). Carl is interested in amphibians and has done field research in Madagascar and

Ecuador. He is currently focusing on the evolution of advertisement calls in frogs, especially

seeking to understand how environmental influences lead to the evolution of distinct calls.

Carl is also interested in the phylogenomics of frogs, and is working to understand anuran

phylogenetic relationships at the Order level, as well as within several families.

Amphib. Reptile Conserv. 144 November 2019 | Volume 13 | Number 2 | e194

Official journal website:

amphibian-reptile-conservation.org

Amphibian & Reptile Conservation

13(2) [General Section]: 145-151 (e196).

New records and distribution extension of the rare glassfrog

Hyalinobatrachium chirripoi (Anura: Centrolenidae)

throughout the Choco-Magdalena region in Colombia

1.2.3.*Angela M. Mendoza-Henao, ‘Roberto Marquez, °Claudia Molina-Zuluaga,

SDaniel Mejia-Vargas, and °Pablo Palacios-Rodriguez

‘Departamento de Zoologia, Instituto de Biologia, Universidad Nacional Autonoma de México, PO 70-153, 04510 Mexico City, MEXICO *Posgrado

en Ciencias Biologicas, Universidad Nacional Autonoma de México, PO 70-153, C.P. 04510, Mexico City, MEXICO *Grupo de Investigacion en

Ecologia y Conservacion Neotropical, 760046, Cali, COLOMBIA *Department of Ecology and Evolution, University of Chicago. 1101 East 57th

St. Chicago, Illinois 60637, USA °Grupo Herpetolégico de Antioquia, Instituto de Biologia, Universidad de Antioquia, A. A. 1226, Medellin,

COLOMBIA °Department of Biological Sciences, Universidad de los Andes, A.A. 4976, Bogota, COLOMBIA

Keywords. Amphibia, Andes Mountains, DNA barcoding, South America, rainforest, range extension

Citation: Mendoza-Henao AM, Marquez R, Molina-Zuluaga C, Mejia-Vargas D, Palacios-Rodriguez P. 2019. New records and distribution extension

of the rare glassfrog Hyalinobatrachium chirripoi (Anura: Centrolenidae) throughout the Chocé-Magdalena region in Colombia. Amphibian & Reptile

Conservation 13(2) [General Section]: 145-151 (e196).

tribution 4.0 International (CC BY 4.0): https://creativecommons.org/licenses/by/4.0/], which permits unrestricted use, distribution, and reproduction in

any medium, provided the original author and source are credited. The official and authorized publication credit sources, which will be duly enforced,

are as follows: official journal title Amphibian & Reptile Conservation; official journal website: amphibian-reptile-conservation.org.

Received: 9 January 2019; Accepted: 16 August 2019; Published: 16 December 2019

Hyalinobatrachium is the most diverse glassfrog genus

(family Centrolenidae) with 32 species described to

date, ranging from Mexico to Argentina (Guayasamin

et al. 2009; Frost 2019). Given the low levels of

morphological differentiation within this genus, species

identification is sometimes difficult, and requires the

use of alternative sources of evidence such as molecular

phylogenetics and DNA barcoding (Castroviejo-Fisher

et al. 2009). Hyalinobatrachium chirripoi (Taylor 1958)

is a seldom observed species found in forests under 600

m elevation from Honduras, along the Choco-Darien

to the Esmeraldas Province in north Ecuador (Kubicki

2007; Guayasamin et al. 2016). Here new records of

H. chirripoi are reported which extend the distribution

of this species into the Andean foothills of the central

Choco and, for the first time, into the Magdalena Valley

of Colombia. An overview of the known distribution

H. chirripoi 1s presented, including previous museum

records and the new data.

Specimens examined. One individual was collected in

2010 at Vereda El Porton, in San Francisco, Antioquia,

Colombia (5.9015, -74.96925, 589 m asl; Fig. 1), in the

Magdalena River drainage. The individual was found

at night, calling on the underside of a Heliconia leaf

overhanging a small stream in a secondary forest, and

was euthanized with an overdose of 2% Roxicaine and

fixed in 10% formalin. A liver sample was preserved in

99% ethanol. The specimen was deposited in the Museo

de Herpetologia, Universidad de Antioquia, Colombia

Correspondence. * am.mendozah@gmail.com

Amphib. Reptile Conserv.

(voucher MHUA-A 06650; originally misidentified

as H. fleischmanni). In 2016, two other individuals

were collected during nocturnal surveys performed

at 20 km SW of Condoto, Choco, Colombia, on the

western versant of the Cordillera Occidental (5.02078,

-76.51633, 423 m asl; Fig. 1A). They were found calling

from the upper side of leaves in a tree hanging above a

small river (Fig. 1B). They were captured and euthanized

with an overdose of topical lidocaine hydrochloride

(Xylocaine). Muscle samples were stored in 100%

ethanol. Specimens were then fixed with 100% ethanol

and deposited in the herpetology collection of the Natural

History Museum at Universidad de los Andes, Colombia

(vouchers ANDES-A3738 and ANDES-A3739). Other

individuals at the same locality were observed on leaves

and fronds of Araceae, Musaceae (Heliconia), and ferns

(Polypodiaceae), perched ~3—8 m off the ground. Both

localities (Vereda El Porton at the Magdalena River

drainage, and 20 km SW of Condoto in the Choco) are

classified as tropical wet forest biome (bh-T, Holdridge

1964).

Morphological and molecular identification. These

samples were identified as H. chirripoi based on light

dorsal spots, significant webbing between Fingers II and

II, clear parietal peritoneum, bare heart condition (.e.,

iridophores covering all visceral peritonea except for the

urinary bladder and pericardium), tympanum visible,

and a truncate snout in sagittal view (Taylor 1958; Ruiz-

Carranza and Lynch 1998; Savage 2002, Fig. 2). To

December 2019 | Volume 13 | Number 2 | e196

Hyalinobatrachium chirripoi in Colombia

0 125 250 375 500

Museum records

\ re Ki

& New Records

Fig. 1. (A) Localities of museum specimens (black dots) and new records (red dots) for Hyalinobatrachium chirripoi. Coordinates

for specimen IAVH-A-4311 (gray dot) were approximated to the urban area of Rio Guapi, Cauca, since precise coordinates of the

collection site were lacking. (B) Habitat where the ANDES-A individuals were encountered.

further corroborate morphological diagnoses, mtDNA

barcoding was used. DNA was extracted following

Ivanova et al. (2006) for specimens ANDES-A3738

and ANDES-A3739, or using the Thermo Scientific

DNA extraction kit for specimen MHUA-A 06650.

Amplification of 16S (567 bp) and COI (609 bp) loci

was as described by Guayasamin et al. (2008), and

Mendoza et al. (2016; primers from Meyer et al. 2005),

respectively. Purified products were Sanger-sequenced

in both directions. Sequences obtained are deposited in

GenBank under accession numbers MH129045—49.

Sequences of both genes were blasted against the

GenBank non-redundant database using megaBLAST.

COI sequences were also used as input for the BOLD

DNA barcoding system (Ratnasingham and Hebert 2007).

In addition, Kimura-two-parameter (K2P; Kimura 1980)

pairwise distances between sequences of closely related

Hyalinobatrachium species available in GenBank (Table

1) were calculated using MEGA7 (Kumar et al. 2016) and

maximum likelihood and Bayesian mtDNA genealogies

were built with RAxML v.8.2.10 (Stamatakis 2006,

2014), and MrBayes 3.2.2 (Ronquist and Huelsenbeck

2003, Ronquist et al. 2012), respectively.

Maximum likelihood searches used the rapid _hill-

Amphib. Reptile Conserv.

climbing algorithm and 10,000 rapid bootstrap pseudo-

replicates to assess nodal support. In MrBayes two

independent 2,000,000 generation analyses were run,

sampling every 1,000 generations, and with 20% burn-in.

The best models for molecular evolution for the 16S and

for each codon position of the COI gene were selected

using PartitionFinder 2 (Lanfear et al. 2016).

Mitochondrial sequences unambiguously confirmed

the identity of the specimens as H. chirripoi. All

BLAST searches against GenBank returned H. chirripoi

sequences as the top hit, with 99% identity and e-values of

zero. Online BOLD identification searches matched the

COI sequences to H. chirripoi with 99.3—99.5% identity.

On the other hand, H. fleischmanni sequences matched

the query sequences with 83.8% similarity (BOLD) and

83.6% identity (GenBank). Maximum likelihood and

Bayesian trees corroborated these results, with the query

sequences nested within a well-supported clade that

includes all the other H. chirripoi (Fig. 3). Finally, K2P

distances among H. chirripoi samples averaged 0.006

(range = 0.002-0.009) for 16S and 0.027 (O—0.039) for

COI, while the mean distance with H. colymbiphyllum,

its sister species, was 0.021 (0.018—0.024) for 16S and

0.081 (0.059-0.093) for COI.

December 2019 | Volume 13 | Number 2 | e196

Mendoza-Henao et al.

Fig. 2. Dorsal (a) and ventral (b) view of live specimen from Chocé-Darien (ANDES-A-3738). Dorsal view (c) of specimen collected

a =

in Madgalena basin (MHUA-A-6650). Details of hand webbing of specimens collected in Choc6-Darien (d) and Madgalena basin (e).

Distribution and conservation implications. The

records of H. chirripoi since its rediscovery by Kubicki

(2004) are very scarce. In Colombia there have been very

few isolated records of the species (Hayes and Starret

1980; Romero-Martinez et al. 2008; Ruiz-Carranza and

Lynch 1998). Most previous records for the species in

Colombia are restricted to the Northwest Choco-Darien

region close to Panama, in Nuqui (MHUA-A 5150-53),

and in Bahia Solano (ICN 40270-314) (Fig. 1). One

additional specimen was collected in 1987 further south,

near Rio Guapi, Cauca (specimen [AVH-Am-4311) with

no georeferenced locality (gray circle in Fig. 1) and the

southernmost specimens were collected from Esmeraldas

Provinces in Ecuador (QCAZ-A 48271, QCAZ-A 66603,

Guayasamin et al. 2016). The new records reported

here fill the gap in the Choco-Darien between the Bahia

Solano and Rio Guapi records, extending the distribution

of this species 70 km into the Chocoan mainland and into

the foothills of the Western Andes (400 m asl).

These records also extend the distribution of H.

chirripoi into the Magdalena basin, across the Andes

from all previous records of this species. Previously, the

closest record of H. chirripoi to the Magdalena basin

was from the Cerro Murrucucu in Tierralta, Cordoba,

Amphib. Reptile Conserv.

within the Parque Nacional Natural Paramillo (ICN

39129-30), an intermediate zone between Choco-Darien

rainforests and Magdalena basin. This region is included

in the Sinu-San Jorge District, characterized by a biota

with common elements, including several amphibian

species of the Chocoan, Amazonian, and Magdalenian

regions (Henao-Sarmiento et al. 2008; Hernandez-

Camacho et al. 1992a; Marquez et al. 2017; Romero-

Martinez et al. 2008; Vasquez and Serrano 2009), and

is considered as a transition zone between the Choco-

Darien, Caribbean, and Magdalena bioregions (Romero-

Martinez et al. 2008). Congruently, the Magdalena basin

record reported here lies within the Nechi District, for

which the biological elements have affinity with those

from the upper Sint and high San Jorge drainages, as

well as the Choco-Darien region (Hernandez-Camacho

étal. 9926).

With these new records of H. chirripoi, the

Magdalena basin and Choco-Darién regions in Colombia

share a total of five species of the genus (including H.

fleischmanni, H. colymbiphyllum, H. aureoguttatum, and

H. valerioi). Hyalinobatrachium aureoguttatum and H.

valerioi are easily differentiable by the dorsal coloration

(large yellow round spots on a green background), but

147 December 2019 | Volume 13 | Number 2 | e196

Hyalinobatrachium chirripoi in Colombia

H. bergeri MHNC 5676

H. pellucidum MNCN 45955

H. aff. pellucidum MAR 2195

1/100

H. esmeralda LSB 384

1/100

H. anachoretus CORBIDI 10462

1/100

H. anachoretus CORBIDI 10472

0.6/10 1/40

0.86/50

1/100

0.02

H. colymbiphyllum KRL 0756

H. colymbiphyllum CH 6822

1/100

H. colymbiphyllum CH 6829

ita H. chirripoi CRAC1005 CR

H. chirripoi CRAC1013 CR

H. chirripoi UCR 17424 CR

D7 /80

H. chirripoi USNM 538586 HN

H. chirripoi AJC 1841 PA

0.8/100 H. chirripoi MHUA A 6650 CO

H. chirripoi ANDES A3738 CO

0.9

-H. chirripot ANDES A3739 CO

Fig. 3. Phylogenetic positions of three Hyalinobatrachium chirripoi samples from Choco and Antioquia, Colombia, in a Bayesian

mtDNA tree inferred from 16S rRNA and COI sequences. The chosen models of evolution using Partition finder were: 16S: GTR+I,

COI position 1: GTR+I, position 2: SYM+G, and position 3: F81+I. Samples in bold are from this study, while the others are

from GenBank. Posterior probability and bootstrap support values (from a maximum likelihood analysis) are indicated in front

of the corresponding node as PP/Bootstrap. Two letter country codes provided for H. chirripoi samples follow the International

Organization for Standardization: CO = Colombia, PA = Panama, HN=Honduras, CR = Costa Rica.

misidentification is common for the other three species

(Kubicki 2004). The most relevant external feature for

differentiating H. chirripoi is the extensive webbing

between Fingers II-III] (H. colymbiphyllum and_ H.

fleischmanni have little webbing between Fingers

II-III); additionally H. fleischmanni has _iridophores

covering the pericardium, while H. chirripoi and H.

colymbiphyllum \ack iridiophores in the pericardial

peritonea (Savage 2002; Starret and Savage 1973, but

check Cisneros-Heredia and McDiarmid 2007). After a

detailed revision of the H. fleischmanni specimens for the

Magdalena basin stored in the Museo de Herpetologia

of Universidad de Antioquia, no additional misidentified

H. chirripoi were found. However, this work highlights

the importance of carefully inspecting museum

specimens of Hyalinobatrachium (and other taxa with

Amphib. Reptile Conserv.

low morphological differentiation between species)

when using such specimens for biogeographic and

conservation work, in order to avoid errors associated

with misidentification.

A shortage of information still remains on the

amphibian diversity in Choco-Darien rainforest and

Magdalena basin, both of which are increasingly

threatened by human activities such as mining, habitat

loss, fragmentation, and other forms of landscape

transformation (Etter and van Wyngaarden 2000; Rangel

2004). Indeed, according to the IUCN Red List, certain

populations of H. chirripoi in Panama and Colombia are

threatened by habitat loss, due to increasing agricultural

activity and logging (Solis et al. 2008). The new records

presented here provide additional information about

the distribution of this rare species, and highlight the

December 2019 | Volume 13 | Number 2 | e196

Mendoza-Henao et al.

Table 1. Sequences for mitochondrial regions 16S and COI of

Hyalinobatrachium chirripoi and related species used in this

study.

Species Voucher 16S COI

H. chirripoi ANDES-A3738 MH129045 MH129047

H. chirripoi ANDES-A3739 MH129046 MH129048

H. chirripoi MHUA-A-6640 MH129049 NA

H. chirripoi UCR 17424 EU663037 NA

H. chirripoi USNM 538586 EU663038 NA

H. chirripoi AJC 1841 KF604299 KF604294

H. chirripoi CRAC1005 NA KJ703 104

H. chirripoi CRACI013 NA KJ703105

H. bergeri MHNC 5676 EU663033 NA

H. pellucidum MNCN 45955 KM068262 NA

H. esmeralda LSB 384 KP149361 KP149161

H. colymbiphyllum KRL 0756 FJ784359 NA

H. colymbiphyllum CH 6829 KR863254 KR862999

H. colymbiphyllum CH 6822 KR863256 KR863001

H. anachoretus CORBIDI 10462 KM068268 NA

H. anachoretus CORBIDI 10472 KM068300 NA

H. aff. pellucidum MAR-2195 KM068296 NA

importance of using an integrative taxonomic approach

at the junction between these two bioregions in terms

of biodiversity conservation, as well as the need for

continued documentation of their biological richness.

Acknowledgements.—This research _is___ partially

supported by UNAM PAPIIT: 203617, by National

Geographic Society Young Explorer’s grant (No. 9786-

15), and by the Rufford Foundation (Rufford Small

Grant reference 18423-1). AMM was supported by

a scholarship from Consejo Nacional de Ciencia y

Tecnologia (CONACyT, Mexico), through Posgrado de

Ciencias Biologicas, Universidad Nacional Autonoma de

Mexico (UNAM). Collections were authorized by permit

No. IBD0359-Res 1117-2014 from the Colombian

Environmental Licensing Authority (ANLA). Fieldwork

in the Magdalena was part of the wildlife characterization

inside the Environmental Impact Assessment (EIA) of

the Samana Norte Hydroelectric exploitation project,

funded by Integral Ingenieria de Consulta S.A and

the specimen collection was conducted under permit

No. 134-0074 granted by the Corporacién Autonoma

Regional de las Cuencas de los rios Negro y Nare -

CORNARE. We thank Mailyn A. Gonzalez and Eduardo

Tovar Luque from Instituto Alexander von Humboldt

(Bogota, Colombia) for access to laboratory facilities;

Andrew J. Crawford, Yiselle Cano, and Luis Alberto

Farfan (Universidad de los Andes, Colombia) facilitated

specimen deposition and access at the ANDES museum;

Mauricio Rivera-Correa provided the images of the

hands of the specimen from Magdalena basin; and Henry

Amphib. Reptile Conserv.

Agudelo-Zamora provided the images from specimens

deposited at Instituto de Ciencias Naturales, Facultad

de Ciencias, Universidad Nacional de Colombia. We

thank Celsa Sefiaris and Jesse Delia for their invaluable

comments to earlier versions of this manuscript, and

Juan M. Daza (Universidad de Antioquia, Colombia)

for access to Museo de Herpetologia Universidad de

Antioquia (MHUA) and his invaluable support in the

development of this manuscript.

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December 2019 | Volume 13 | Number 2 | e196

Amphib. Reptile Conserv.

Mendoza-Henao et al.

Angela M. Mendoza-Henao is a biologist from Universidad del Valle (Colombia) with an M.S.

in Biological Sciences, and a current Ph.D. student at the Universidad Nacional Autonoma de

México (UNAM). Angela’s work has focused mainly in the application of molecular tools for

solving questions in ecology and conservation, with an emphasis on terrestrial vertebrates, mainly

neotropical amphibians.

Roberto Marquez is a Ph.D. candidate in Ecology and Evolution at the University of Chicago

(Chicago, Illinois, USA). Roberto’s research is mostly focused on evolutionary genetics,

systematics, development, and behavior of Dendrobatid poison frogs. He earned B.Sc. and M.Sc.

degrees from Universidad de los Andes in Bogota, Colombia.

Claudia Molina-Zuluaga is a biologist from Universidad de Antioquia (Colombia), with an MLS. in

Biological Science from Universidad Nacional Autonoma de México (UNAM). Claudia’s research

is mostly focused on the population ecology of amphibians and reptiles, using a demographic

methods approach to population dynamics in order to establish long-term trends and conservation

status.

Daniel Mejia-Vargas is a biologist doing independent research on the systematics, biogeography,

and behavior of poison frogs. Daniel is also involved in ethnobiology research and studies diversity

in plants, fishes, amphibians, and birds.

Pablo Palacios-Rodriguez is a biologist working on his Ph.D. in Biological Sciences at Universidad

de los Andes (Colombia). Pablo is interested in the evolution of toxicity, the discovery of new

species, and the physiology of the personality of frogs.

151 December 2019 | Volume 13 | Number 2 | e196

Official journal website:

amphibian-reptile-conservation.org

Amphibian & Reptile Conservation

13(2) [General Section]: 152-159 (e198).

Natural history notes on three sympatric frogs, Amolops

formosus (Gunther 1875), Nanorana liebigii (Gunther 1860),

and Ombrana sikimensis (Jerdon 1870), from Manaslu

Conservation Area, Nepal

'*Biraj Shrestha and 7Min Bahadur Gurung

'SAVE THE FROGS!, 1968 South Coast Hwy Suite 622, Laguna Beach, California 92651, USA *Small Mammals Conservation and Research

Foundation, Lalitpur, NEPAL

Abstract.—Three stream dwelling mountain frogs, Amolops formosus, Nanorana liebigii, and Ombrana

sikimensis are sympatric species, native to Asia and distributed much across Nepal. Here, a brief natural

history account of the three species is provided that enhances the existing knowledge of these understudied

frogs. Altogether 21 adults (eight Amolops formosus, six Nanorana liebigii, and seven Ombrana sikimensis)

were collected from the streams of Sirdibas, Chumchet, and Bihi villages in April and May 2016 and in March

2017. Since the survey time coincided with breeding season, egg clutches and tadpoles of Nanorana liebigii

were observed. Basic morphometric features of the adults (snout-vent length, head length, head width,

femur length, and tibia length) and tadpoles (total length, body length, body width, and tail muscle width)

were measured with a Mitutoyo digital Vernier caliper to the nearest 0.1 mm. Environmental parameters of

the habitat were also noted, including air temperature, water temperature, relative humidity, and pH of the

water body. A review of the conservation status of these sympatric frogs highlights the threats they face from

unchecked harvesting in Manaslu and across the entire mountain villages of Nepal. Other potential threats

include declining stream habitats through water use management decisions such as dams and diversions,

pollution, and forest degradation. The field observation data collected will help to fill in the knowledge gaps for

these species, in order to prioritize conservation action and aid future research.

Keywords. Amphibia, Anura, Asia, habitat degradation, morphometrics, threats

Citation: Shrestha B, Gurung MB. 2019. Natural history notes on three sympatric frogs, Amolops formosus (Gunther 1875), Nanorana liebigii

(Gunther 1860), and Ombrana sikimensis (Jerdon 1870), from Manaslu Conservation Area, Nepal. Amphibian & Reptile Conservation 13(2) [General

Section]: 152-159 (e198).

tribution 4.0 International (CC BY 4.0): https://creativecommons.org/licenses/by/4.0/], which permits unrestricted use, distribution, and reproduction in

any medium, provided the original author and source are credited. The official and authorized publication credit sources, which will be duly enforced,

are as follows: official journal title Amphibian & Reptile Conservation; official journal website: amphibian-reptile-conservation.org.

Received: 16 August 2018; Accepted: 25 July 2019; Published: 11 November 2019

Introduction

Amolops formosus, Nanorana liebigii, and Ombrana

sikimensis are sympatric species that are largely dependent

upon mountain brooks and associated riparian habitats

characterized by coniferous or oak forests (Schleich and

Kastle 2002). They are native to Asia, found across many

of the mountains of Nepal, and also recorded in India,

China, Bangladesh, and Bhutan (Bordoloi et al. 2004;

Liang et al. 2004). In Nepal, they are distributed within

an altitudinal range of 1,190-3,360 m asl (Schleich

and K4astle 2002). All three species were previously

placed in the genus Rana (Boulenger 1920), but later

revised into distinctive genera of Amolops, Nanorana,

and Ombrana (Chen et al. 2005; Dubois 1974; Frost

et al. 2006). The earliest first-hand records related to

morphometrics, life history, and habitat notes of the three

species (Boulenger 1882; Giinther 1860; Jerdon 1870)

are not readily accessible at the present time. While the

recent publication of Schleich and Kastle (2002) is rather

comprehensive, it is still unavailable to many readers due

to the high price of the book (Zug 2004). Shah and Tiwari

(2004) provided little information on the associated

habitats of these sympatric amphibians, like surrounding

vegetation and environmental parameters, but their report

lacks data on egg deposition and tadpole stages. The

IUCN Red List Assessment 2004 has further emphasized

the need for research on the taxonomy, population size,

distribution, trends, ecology, and life history of these

frogs to prioritize conservation actions (Bordoloi et al.

2004; Liang et al. 2004). Therefore, any readily-available

publication on the natural history of these frogs is of great

importance to the scientific community, conservationists,

natural resource managers, and decision makers.

Stream-dwelling frogs serve as good indicators of

the stream ecosystem health, since they are philopatric

Correspondence. * thepristinewoods@gmail.com, biraj@savethefrogs.com; 7 tamumin23@gmail.com

Pp Ss Y S S

Amphib. Reptile Conserv.

November 2019 | Volume 13 | Number 2 | e198

Shrestha and Gurung

Location Map of Manaslu Conservation Area

28°30'0"N 28°40'0"N 28°50'0"N

28°20'0"N

4:1,500,000. ~

84°30'0"E

y 2 > ¥ - ‘ & = =

1:580,000 - pt, a

Coordinate System: GCS WGS 1984 . ;

Datum: WGS 1984 ‘

Units: Degree

as ee

34°40°0"E 84°50'0"E 85°10°0"E

Legend = Village Boundary ---- Road Network z= Manaslu Conservation Area Boundary

Fig. 1. Study sites in Manaslu Conservation Area, Nepal, with villages indicated in blue text.

in nature and found in steady populations (Welsh and

Ollivier 1998). Studying such stream frogs with respect

to morphology, life history, and habitat conditions will

help to further understanding of their ecological niches

(Ningombam 2009). This knowledge is vital for devising

efficient conservation strategies when one-third of the

total amphibian species of the world are being threatened

with extinction (Baillie et al. 2004). This study presents

the natural history notes of these three sympatric frogs.

This information will help to aid in further research and

monitoring, while providing background support for

good decision making regarding their conservation in the

future.

Materials and Methods

Surveys. Manaslu Conservation Area is one of the

protected areas in Nepal, located at the upper north

area of Gorkha district, province number 4 (Gandaki

Pradesh). Surveys were conducted in five major villages

of the Manaslu Conservation Area, namely Sirdibas,

Bihi, Chumchet, Prok, and Samagaun, excluding Lho

and Chhekampar (Fig. 1). The entire survey spanned 49

days during the day-time in April-May 2016 and March

2017.

A distance of 279 km was covered on foot throughout

the survey and a transect of 200 m was walked in each

site of the 14 streams (Table 1). Time-constrained

searches were conducted for 2 h with two people at

a time, for a total of four person-h per search. Live

specimens of Amolops formosus, Nanorana liebigii, and

Ombrana sikimensis were collected for morphological

Amphib. Reptile Conserv.

examination. They were released in-situ after recording

observational notes and taking photographs with a Canon

EOS 700D (18-135 mm) kit lens DSLR camera (Fig. 2).

Egg clutches and tadpoles of different sizes were found

in a few streams, and they were closely observed while

causing minimal disturbance. The distribution of these

sympatric frogs in Manaslu is restricted to Sirdibas, Bihi,

and Chumchet (Fig. 3).

Measurements. The snout-vent length (SVL), head

length (HL), head width (HW), femur length (FL), and

tibia length (TL) were the morphological parameters

measured following Fei et al. (2009) for the adult frogs.

The morphometric keys for tadpoles were total length

(TL), body length (BL), body width (BW), and tail muscle

width (TMW), and followed Mitchell et al. (2012). All

the measurements were taken using a Mitutoyo digital

Vernier caliper to the nearest 0.1 mm.

Air and water temperature measurements were taken

using a digital thermometer. The humidity was measured

using a Hygrometer and the pH of the water was

recorded with a digital pH meter. Geographic coordinates

and altitude were recorded with a Garmin eTrex 10

GPS. Species identification and additional information

followed Boulenger (1882, 1920), Chen et al. (2005),

Frost et al. (2006), Gunther (1860), Ningombam (2009),

Schleich and Kastle (2002), and Shah and Tiwari (2004).

Results and Discussion

Morphometrics. The morphological notes of the

three sympatric frogs correspond well with the earlier

November 2019 | Volume 13 | Number 2 | e198

Three sympatric frogs in Manaslu Conservation Area, Nepal

Table 1. Sampling locations and observations of frogs (adults, egg clutches, and tadpoles) in the survey.

Sampling Site

ies)

on nN N

Village

Sirdibas

Chumchet

Samagaun

Prok

Bihi

Location

Yuwang Khola*

Ghatte Khola

Myarchwang

Khola

Gyanak Khola

Sipchet Ripchet

Chumling

Gumlung Khola

Sardi Khola

Lokpa

Phujung Khola

Birendra Tal

Namrung Khola

Bihi Khola

Dyang Khola

Altitude (m asl) Survey Time

1,622

2,425

1,629

2,294

2,473

2,485

2,482

1,938

1,887

1,931

3,700

2,462

2,189

1,838

*K hola refers to stream

descriptions provided by Giinther (1860, 1875) and

Jerdon (1870) quoted in Schleich and Ka4stle (2002).

Nanorana liebigii has the largest mean body size and body

weight followed by Ombrana sikimensis and Amolops

formosus. The body weight measurements of these

frogs found in Nepal did not set new records. In Bhutan,

Wangchuk (2017) documented the average weight of

Nanorana liebigii as exceptionally higher (males 500—

750 g and females 350-500 g) than the present findings

for Nepal. The head lengths (HL) of Amolops formosus

and Nanorana liebigii were smaller than the head widths

(HW), however, Ombrana_ sikimensis was different,

with HL greater than HW (Table 2). On the contrary,

Boulenger (1920) has described broader HW than HL in

Ombrana sikimensis.

Frogs in general tend to exhibit sexual dimorphism,

with females mostly being larger in body size than males

(Monnet and Cherry 2002). The results here agree for

Amolops formosus, where females were larger in size

than males; however, the males of Nanorana liebigii were

Observations

Day Amolops formosus (2 2, 1 4); Nanorana

liebigii (5 tadpoles, egg clutch)

Day None

Day Nanorana liebigii (1 4); Ombrana

sikimensis (7 individuals, unidentified

sex)

Dawn Amolops formosus (3 9); Nanorana

liebigii (1 3)

Day Nanorana liebigii (1 3)

Dawn Nanorana liebigii (1 )

Day None

Dawn Nanorana liebigii (tadpole with

metamorphosed legs)

Day Nanorana liebigii (egg clutch)

Day Nanorana liebigii (1 3)

Day None

Day Amolops formosus (1 9°); Nanorana

liebigii (1 3)

Night Amolops formosus (1 9)

larger in size than their female counterparts. The adult

males of Amolops formosus had a nuptial pad on the 1*

finger of the forelimb and also partly turquoise-colored

hind limbs on the ventral side (Fig. 4A), as documented

by Schleich and Kastle (2002). Likewise, males of the

Nanorana liebigii had strongly hypertrophied forelimbs

with a nuptial pad on the 1* finger. Further, numerous

black horny spines were present on the 1°, 2" and 3”

fingers on both the arms and extending along the pectoral

region (Fig. 4C). It was difficult to identify the sexes of

Ombrana sikimensis based only on SVL measurements,

since nuptial spines are not present in Ombrana sikimensis

(Boulenger 1920).

Egg deposition and larval stages. Spring (March—

May) is the season of breeding for Nanorana liebigii

as egg clutches were found in slowly drifting Yuwang

Khola in Sirdibas village to fast flowing streams in

Lokpa, Chumchet village. The eggs were attached to the

undersides of stones, totally submerged, and white in

Table 2. Morphological parameters (SVL, HL, HW, FL, and TL) of the three species of adult sympatric frogs (mm) and BW (g). Min

= Minimum value, Max = Maximum value, M = Average value (Mean), SD = Standard Deviation, and n = number of individuals.

Amolops formosus

Morphometric keys (n= 8)

Min M+SD Max Min

Snout-vent length(SVL) 67.3 74.14+3.8 81.5 78.8

Head length (HL) 230. “2a hr 26-1 25.8

Head width (HW) 246 258410 27.9 274

Femur length (FL) 399 429419 461 43.2

Tibia length (TL) 452 475412 49.1 47.6

Body weight (BW) 35 46.9+6.2 55 60

Amphib. Reptile Conserv. 154

Nanorana liebigii Ombrana sikimensis

(n= 6) (n=7)

M+SD Max Min M+SD Max

87.74£7.6 99.6 67.1 81.24 11.6 92.1

27. 9+.2:0 31.5 21.6 24.6°£:2:3 27.1

29.1+1.8 32.4 20.5 233-4592 25.8

50.6 + 3.7 53.1 39.3 44.0+3.9 47.9

54-7439 58.7 42.3 46.44 3.4 50.2

82.84 148 100 50 70.1 + 14.6 85

November 2019 | Volume 13 | Number 2 | e198

Shrestha and Gurung

ite? "ee Ae

Fig. 2. Dorsal view of live adults: (A) Ombrana sikimensis, (B) Amolops formosus, and (C) Nanorana liebigii. (D) Dorso-lateral

view of Nanorana liebigii. Photos: Biraj Shrestha and Min Bahadur Gurung.

color inside the gelatinous ball that had a honeycomb-

like appearance (Fig. 5A—B). The clutch size consisted

of about 80-140 eggs although no adults were seen

nearby, which is noteworthy since males of Nanorana

liebigii are reported to guard the eggs as a form of

parental care (Rai 2003). Some gradually developing

egg clutches were observed that had embryos with eyes

and were surrounded by jelly of a “liver-like” color

(Fig. 5C).

Five tadpoles of Nanorana liebigii were observed at

the Yuwang Khola and one metamorphosed tadpole with

hind limbs was evident in the shallow pools of a rapidly

flowing stream in Sardi Khola. The metamorphosed

tadpole had a well-developed oral disc (Fig. 5D), with the

structure of the upper and lower lips forming an atrium

feature (Kastle et al. 2013). The tails of the tadpoles were

nearly twice the length of the body (Schleich and Kastle

2002), while the body lengths were significantly greater

than widths (Table 3).

Tadpoles of Nanorana liebigii were found sympatric

with the tadpoles of Duttaphrynus himlayanus. However,

no egg clutches of Amolops formosus were observed

during the study. Published information on egg deposition

by Amolops formosus is limited, though Nidup et al.

(2016) reported an egg clutch of Amolops himalayanus

attached underneath of rocks and clear white, from a

gentle flowing stream in Bhutan.

Habitat notes. The general habitat ofall the three sympatric

frogs studied here is mountain streams above 1,100 m asl

Amphib. Reptile Conserv.

elevation and of varied intensity. In addition, Nanorana

liebigii was also found to inhabit other water bodies, such

as the puddles in a bamboo forest and irrigation ditches

near the cropland where Karu (a type of naked barley)

was grown (Fig. 6). Amolops formosus preferred fast

flowing streams, typically cascades, attaching themselves

to the steep slopes of rocks and resting on fissures, partly

covered in moss and ferns, while Ombrana_ sikimensis

were typically hiding in clusters underneath rocks in

shallow streams (Fig. 6C). The nearby riparian vegetation

included Nepalese Alder (Alnus nepalensis), Broom

Grass (Thysanolaena maxima), Himalayan Blue Bamboo

(Himalayacalamus hookerianus), Himalayan Silver

Birch (Betula utilis), Tree Rhododendron (Rhododendron

arboreum), Chir Pine (Pinus roxburghii), Walnut (Juglans

Table 3. Morphological parameters (TL, BL, BW, and TMW)

of the tadpoles of Nanorana liebigii (mm). Min = Minimum

value, Max = Maximum value, M = Average value (Mean), SD

= Standard Deviation, and n = number of individuals.

Nanorana liebigii

Morphometric keys Tadpole (n = 6)

Min M+SD Max

Tail length (TL) 414 491+6.7 577

Body length (BL) 13.3 193445 245

Body width (BW) So. GMEG 5 14.7

Tail muscle width (TMW) one Fpeie asl) 1g bs;

November 2019 | Volume 13 | Number 2 | e198

Three sympatric frogs in Manaslu Conservation Area, Nepal

Amphibian Occurrence and Distribution in Manaslu Conservation Area, Gorkha District, Nepal

Samagaun | a

Tee t _

Clq SSere-k Sam

28°39'0"N

28°32'0"N

pani Peak (Lidanai Pefk)

Luksui

1:300,000

Apu aQhung ae

aye ree

f aoe pBaibhuk

( Cy Asyarangee”enakhar,

Chumchet \

Seyjae ee aes ¢ (

car i

a Pied ( \ f

Rs ‘

~ gs ae Y Pp x

Chhilungkholagaun

28°25'0"N

Ghyachok

® Amolops formosus @ Nanoranaliebigii @

Legend % Duttaphrynus himalayanus

84°30'°0"E

28°18'0"N

_Sirdibas.,.

ae e aatuaees S

84°50°0"E

Ores .)

Masing Now

eWlenore’f

Nagjet 2

sa, Siribas’ my

Local Village Point 4 Metamorphosed Tadpole

Transect

85°10°0"E

Fig. 3. Frog observation sites and distances travelled throughout the survey.

regia), and others.

Amphibians are often mentioned as the bio-indicators

of water quality because of their permeable skin.

Their eggs and larval stages are generally much more

vulnerable to any type of pollution in the water bodies due

to their lack of protective covering, and direct connection

with the water for survival and growth. The optimal

environmental conditions of their habitat are crucial to

allow for metamorphosis and for habitat management

strategies. Cold water is favorable for the growth of

different life stages with neutral to slightly alkaline water

pH. This range is desirable for much of the aquatic fauna,

including stream dwelling frogs, as lower acidic pH

conditions can impede amphibian growth and inhibit the

development of eggs and embryos (Ningombam 2009).

Conservation status. All the three species of frogs are

listed in the Least Concern (LC) category in the IUCN

Red List Assessment from 2004 (15 years prior to this

writing) based upon the presumption of their large

populations, wider distributions, and with no prospects

of immediate decline. However, a reassessment 1s

desperately needed, as the current population trends for

all three species are going down due to declining stream

habitats from various causes, such as water diversion

and dams to deforestation and pollution (Bordoloi et

al. 2004; Liang et al. 2004). In India, all three of these

species are protected under the national legislation, while

no similar effort by the Nepalese government has been

undertaken to provide any legal measures for amphibian

conservation. This remains the case today (in 2019),

despite the recommendation by the Biodiversity Profiles

Amphib. Reptile Conserv.

Project (BPP, 1995) for nine species of endemic Nepalese

amphibians to be included in the Schedule I of National

Parks and Wildlife Conservation (NPWC) Act 1973.

Anurans of genera Amolops, Nanorana, and Ombrana

are called ‘Paha’ frogs in Nepal and have ethnozoological

relationships with the communities living mostly in hills

and mountains (Shah and Tiwari 2004; Shrestha 2018).

People often harvest paha frogs as a delicacy and for

their apparent therapeutic benefits. Every indigenous

community in the mountains of Nepal either has

experience in paha hunting or at least knows about its

use. As a result, paha hunting is popular in villages from

the east to western part, all across the nation. The hunting

usually takes place at night during pre- and post-monsoon

seasons, when the water flow is minimum. There is

no limit for harvested quantities from the streams, and

people usually collect as many as they can find during

their searches. In Manaslu, Gurung communities in

Sirdibas village typically collect 51-100 individuals on

Table 4. Physico-chemical characteristics of water quality in

the survey sites and altitudinal range of the detected frogs. Min

= Minimum value, Max = Maximum value, M = Average value

(Mean), and SD = Standard Deviation.

Abiotic factors Min M+ SD Max

Air Temperature (°C) 8 20.0 + 6.5 26.5

Water Temperature (°C) 4 132-22. 8 16.3

Relative Humidity (%) 25 ASS e117 55

pH cs 8.0+0.3 8.6

Altitude (m asl) 1,591 1,880.5+439.4 2,480

November 2019 | Volume 13 | Number 2 | e198

Shrestha and Gurung

and (D) female of Nanorana liebigii. Photos: Biraj Shrestha.

average in one season and they trade locally in the price

range of USD 0.45—2.26 (Shrestha and Gurung 2019).

Nanorana liebigii is the most popular paha frog across

the country, followed by Ombrana sikimensis which

is highly sought after as its meat serves as a delicacy

and nutritional purposes (Shah and Tiwari 2004). In

addition, the meats of Nanorana liebigii and Amolops

formosus are traditionally assumed to have medicinal

properties that cure fever, cough, cold, dysentery, and

stomach ache; while their skin secretions have antiseptic

properties (Shrestha and Gurung 2019). In recent years,

paha hunting 1s largely practiced to enjoy its meat and for

recreational purposes in the villages, since the nutritional

requirements are often met by poultry and livestock, and

medical supplies are readily available thanks to improved

road access for most villages these days. However, the

continued paha hunting practice has depleted its numbers

as reported by the local communities across the country,

including Manaslu, and has led to the recommendation

for some form of legal conservation protection.

The diminishing populations of Amolops formosus,

Nanorana liebigii, and Ombrana_ sikimensis can be

averted by developing species specific conservation

priority plans. Habitat conservation planning, population

study and monitoring, hunting regulation policies, and

effective outreach programs are some of the key action

steps. The brief natural history notes presented here on

morphometrics, sexual dimorphism, egg deposition,

larval stages, and habitat conditions will be helpful in

this regard. But since the identification of congeneric

amphibians can be tricky, the use of molecular phylogeny

Amphib. Reptile Conserv.

and call identification coupled with morphometrics is

strongly recommended for accurate species identification.

Acknowledgements.—We are thankful to the Rufford

Foundation, UK for funding this research in the first

place. Then, we acknowledge the following institutions

and individuals; SAVE THE FROGS!, Friends of

Nature (FON) Nepal, Department of National Parks and

Wildlife Conservation (DNPWC), National Trust for

Nature Conservation (NTNC) Manaslu Conservation

Area Project (MCAP) Office, Gorkha and Philim, The

Pollination Project (TPP) USA for their technical input

and facilitating this study, approval of the research

permits and support through additional funding. We also

thank Saney Pd Suwal and Mano) Konga who assisted

with the field works, local community of Manaslu for

embracing our mission to protect the paha frogs, Bishnu

Maharjan for producing the GIS maps and Kiran Lohani

for the media exposure. Finally, we value the feedback

received from all the anonymous reviewers and the

journal editorial team (ARC).

Literature Cited

Baillie JEM, Hilton TC, Stuart SN. 2004. JUCN Red List

of Threatened Species. A Global Species Assessment.

International Union for Conservation of Nature, Gland,

Switzerland and Cambridge, United Kingdom. 191 p.

Bordoloi S, Ohler A, Shrestha TK. 2004. Ombrana

sikimensis. The IUCN Red List of Threatened Species

2004: e.T58246A 11757068.

November 2019 | Volume 13 | Number 2 | e198

Three sympatric frogs in Manaslu Conservation Area, Nepal

Fig. 5. Life sta es of Nanorana liebigii: (A) eggs deposition underneath a stone, (B) eg: clutch, (C) embryo development, and (D)

Fig. 6. Habitat varieties of the three sympatric frogs: (A) rapidly flowing stream, (B) series of waterfalls inhabited by Amolops

formosus, (C) slow flowing shallow stream, and (D) irrigation ditch. Photos: Biraj Shrestha.

Amphib. Reptile Conserv. 158 November 2019 | Volume 13 | Number 2 | e198

Shrestha and Gurung

Bordoloi S, Ohler A, Shrestha TK, Ahmed MF. 2004.

Amolops formosus. The IUCN Red List of Threatened

Species 2004: e.T58206A 11746654.

Boulenger GA. 1882. Catalogue of the Batrachia

Salientia s. Ecaudata in the Collection of the British

Museum. Second Edition. Taylor and Francis, London,

United Kingdom. 503 p., 30 pls.

Boulenger GA. 1920. A Monograph of the South Asian,

Papuan, Melanesian and Australian Frogs of the

Genus Rana. Records of the Indian Museum, Volume

20. Zoological Survey of India, Calcutta, India. 226 p.

BPP [Biodiversity Profiles Project]. 1995. Red Data Book

of the Fauna of Nepal Biodiversity. Profiles Project

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Parks and Wildlife Conservation, Ministry of Forests

and Soil Conservation, His Majesty’s Government of

Nepal, Kathmandu, Nepal.

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Ho T, Somorjai ILM. 2005. Taxonomic chaos in

Asian ranid frogs: An initial phylogenetic resolution.

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au Nepal. Bulletin du Museum National d Histoire

Naturelle 213: 341-411.

Fei L, Hu SQ, Ye CY, Huang YZ. 2009. Fauna Sinica:

Amphibia. Science Press, Beijing, China. 957 p.

Frost DR, Grant T, Fatvovich J, Bain RH, Haas A,

Haddad CFB, de Sa RO, Channing A, Wilkinson M,

Donnellan SC, et al. 2006. The amphibian tree of life.

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297. 1-291.

Gunther ACLG. 1860. Contribution to the knowledge of

the reptiles of the Himalaya Mountains. Proceedings

of the Zoological Society of London 1860: 148-175.

Jerdon TC. 1870. Notes on Indian herpetology.

Proceedings of the Asiatic Society of Bengal 1870:

66-85.

Kastle W, Rai K, Schleich H. 2013. Field Guide to

Amphibians and Reptiles of Nepal. ARCO-Nepal,

Munich, Germany. 609 p.

Liang F, Lau MWN, Dutta S, Shrestha TK, Borah MM.

2004. Nanorana liebigii. The IUCN Red List of

Threatened Species 2004, e.T58428A 11780058.

Mitchell T, Alton LA, White CR, Franklin CE. 2012.

Relations between conspecific density and effects of

ultraviolet-B radiation on tadpole size in the Striped

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Marsh Frog: UVBR and conspecific-density effects

on amphibians. Conservation Biology 26(6): 1,112-

Lal)

Monnet JM, Cherry MI. 2002. Sexual size dimorphism in

anurans. Proceedings of the Royal Society of London

B Biological Sciences. 269: 2,301—2,307

Nidup T, Gyeltshen D, Dorji S, Pearch MJ. 2016. The

first record of Amolops himalayanus (Anura: Ranidae)

from Bhutan. The Herpetological Bulletin 136: 13-18.

Ningombam B. 2009. Amphibian fauna in and around

Loktak Lake, Manipur, India with reference to the

genus Amolops Gunther. Ph.D. Dissertation, Gauhati

University, Jalukbari, Guwahati, Assam, India.

Rai KR. 2003. Environmental impacts, systematics,

and distribution of herpetofauna from east Nepal.

Ph.D. Dissertation, Tribhuvan University, Kirtipur,

Kathmandu, Nepal.

Schleich H, Kastle W. 2002. Amphibians and Reptiles

of Nepal: Biology, Systematics, Field Guide. First

Edition. A.R.G. Ganter Verlag, Ruggell, (Liechtenstein.

k201ep.

Shah KB, Tiwari S. 2004. Herpetofauna of Nepal: A

Conservation Companion. IUCN Nepal, Kathmandu,

Nepal. 237 p.

Shrestha B. 2018. Amphibian Conservation: Brief

Introduction in the Context of Nepal. The Rufford

Foundation, London, United Kingdom; SAVE THE

FROGS!, Laguna Beach, California, USA; and

Resources Himalaya Foundation, Kathmandu, Nepal.

20 p.

Shrestha B, Gurung MB. 2019. Ethnoherpetological

notes regarding the Paha Frogs and conservation

implications in Manaslu Conservation Area, Gorkha

District, Nepal. Journal of Ethnobiology and

Ethnomedicine 15(1): 23.

Wangchuk S. 2017. Morphometric Study of Mon-Paa

Frog: A Case Study of Dopuchen, Dumtoe and Tendruk

Gewog under Samtse Dzongkhag. Dzongkhag

Forestry Sector, Samtse Dzongkhag, Bhutan. 24 p.

Welsh HH, Ollivier LM. 1998. Stream amphibians as

indicators of ecosystem stress: A case study from

California’s redwoods. Ecological Applications 8(4):

1,118—1,132.

Zug GR. 2004. Book Review: Amphibians and Reptiles

of Nepal: Biology, Systematics, Field Guide.

Herpetological Review 35(1): 88-90.

Biraj Shrestha has been affiliated with the California-based amphibian conservation non-profit,

SAVE THE FROGS! since 2013. Biraj obtained a Master’s degree in Environmental Science from

Khwopa College, Tribhuvan University (Kathmandu, Nepal) in 2013. He is deeply interested in

the systematics, evolution, phylogenetics, ecology, ethnobiology, and conservation science of

amphibians. Currently, Biraj is pursuing a Master of Science degree (M.S.) in the Coastal Science

and Policy (CSP) program at the University of California, Santa Cruz, California, USA.

Min Bahadur Gurung is a free-lance researcher and life member of the Small Mammals

Conservation and Research Foundation, Nepal. Min has a Bachelor’s degree from Birendra

Multiple Campus and a Master's degree in Zoology from the Central Department of Zoology,

Tribhuvan University, Kathmandu, Nepal. His research interests include distribution, diversity, and

conservation of amphibians, reptiles, birds, and mammals.

November 2019 | Volume 13 | Number 2 | e198

Official journal website:

amphibian-reptile-conservation.org

Amphibian & Reptile Conservation

13(2) [General Section]: 160-171 (e200).

Checklist of the amphibians and reptiles of the

Lely Mountains, eastern Suriname

Rawien Jairam

National Zoological Collection of Suriname, Anton de Kom University, Paramaribo, SURINAME

Abstract.—The Lely Mountains in Suriname have been surveyed only a few times by various herpetologists

since 1973, and most recently in June 2016 by three people during six days. The total number of species

recorded from the Lely Mountains is 102, including 46 species of amphibians and 41 species of reptiles in the

2016 surveys, with 15 additional species from the previous survey. Pluviometric conditions were not favorable

during the survey, so more species of amphibians likely remained undetected. The use of only active searches

probably only allowed detection of a portion of the reptile fauna of the massif. Unfavorable weather and the

relatively small area sampled indicate that the diversity of the herpetofauna of the Lely Mountains is probably

far from being completely documented. The presence of roads established for illegal gold mining and other

human structures, such as a communication tower, have caused significant forest degradation.

Keywords. Anura, conservation, herpetofauna, Lacertilia, laterite landscape, new records, Serpentes, South America,

Testudines

Citation: Jairam R. 2019. Checklist of the amphibians and reptiles of the Lely Mountains, eastern Suriname. Amphibian & Reptile Conservation 13(2)

[General Section]: 160—171 (e200).

ternational (CC BY 4.0): https://creativecommons.org/licenses/by/4.0/], which permits unrestricted use, distribution, and reproduction in any medium,

provided the original author and source are credited. The official and authorized publication credit sources, which will be duly enforced, are as follows:

official journal title Amphibian & Reptile Conservation; official journal website: amphibian-reptile-conservation.org.

Received: 18 October 2018; Accepted: 15 May 2019; Published: 14 November 2019

Introduction

Presently the herpetofauna of Suriname consists of

approximately 303 species (Ouboter 2017). Although

over 90% of Suriname is still covered by natural

vegetation, only a few studies have assessed the

herpetofaunal diversity of Suriname throughout the

entire territory (Hoogmoed 1973; Ouboter and Jairam

2012) leaving many areas still unexplored. Herpetofaunal

surveys in specific areas have been conducted by several

researchers (Ouboter et al. 2007, 2011; Nielsen et al.

2013; Fouquet et al. 2015a,b). New country records are

still being documented, e.g., Amapasaurus tetradactylus

(Jairam and Jairam-Doerga 2015), and range extensions

continue to be reported, e.g., Ptvchoglossus brevifrontalis

(Jairam and Jairam-Doerga 2016).

The Lely Mountains are located in the eastern part

of Suriname (4°25’-4°45°N, 54°39’-54°55’W) and

together with the Brownsberg, Nassau, Winti Wai, Hok-

a-Hin, Stonbroekoe, and Majordam Mountains they

form a system of laterite-bauxite plateaus in northeastern

Suriname. These small plateaus form a unique type of

landscape (H. ter Steege, pers. comm.) with different

vegetation types, including high forest, savannah forest,

and rocky creek beds (Alonso and Mol 2007). The Lely

Correspondence. rawien_2000@yahoo.com

Amphib. Reptile Conserv.

Mountains together with the Brownsberg and the Nassau

Mountains comprise a bauxite concession of the Suriname

Aluminum company (Suralco), which formerly operated

in Suriname (Mol et al. 2007). Although Suralco explored

the Lely Mountains for bauxite deposits, the company

did not proceed with the mining exploitation. The Lely

Mountains consist of several plateaus with a maximum

height of approximately 700 m (Alonso and Mol 2007).

Due to the absence of established roads for cars, the Lely

Mountains are mainly accessible only by air. A trail that

is used by gold miners going up the Lely Mountains by

means of all-terrain vehicles is long and tedious, and

seldom without dangers. These circumstances have

led to relatively few surveys (Hoogmoed 1974, 1975,

unpub.; Watling and Ngadino 2007) at this location when

compared to other nearby bauxite mountains, such as the

Brownsberg Mountain. The remoteness of this location

may also favor the occurrence of microendemics and/or

rare species such those as in Anomaloglossus (Vacher et

al. 2017). However, information about the herpetofauna

of the Lely Mountains 1s still scant. This report documents

new findings from the herpetofauna surveyed in the Lely

Mountains in June 2016, and presents an overview of the

herpetofaunal diversity observed and an updated list for

this location.

November 2019 | Volume 13 | Number 2 | e200

Rawien Jairam

pueINs) YOUdI4

Legend

— Large creeks

r Rivers

‘(ig Brokopondo Lake

/ ALTITUDE

| j|0

[| 50

100

250

500

750

1000

Fig. 1. Map of the Lely Mountains plateau showing the area surveyed in the red square. The inset image shows the location of the

Lely Mountains in Suriname, as indicated by the red arrow. In the right upper side of the figure the Nassau Mountains are visible.

Materials and Methods

Habitat description. The vegetation and water

drainages in the Lely Mountains are presently heavily

disturbed locally by illegal small-scale gold miners.

Some of the areas sampled were the airstrip, which was

covered with grass, and small ditches located on one

side for water drainage. The area around the airstrip was

surrounded by patches of pristine forest interspersed

with disturbed forests. The vegetation in the disturbed

areas included mostly trees that were approximately

two m high. The cause of this disturbance could have

been the establishment of the airstrip by Suralco. The

Amphib. Reptile Conserv.

bauxite company also established roads that extend from

the plateau to the northern part of the Lely Mountains.

The aforementioned roads were usually dominated by

large water filled potholes bordered by small shrubs and

bushes. Also sampled was a small creek on the plateau,

which was heavily disturbed by artisanal gold mining.

Survey methods. Three people opportunistically

surveyed the forest of the Lely Mountains using available

roads and trails for six days. Trails and roads were surveyed

radiating in different directions from the airstrip (Fig. 1).

The surveyors also walked through the forest away from

trails to increase the chance of finding amphibians and

November 2019 | Volume 13 | Number 2 | e200

Herpetofauna of the Lely Mountains, Suriname

reptiles. Night surveys started at dusk, at approximately

19.00 h, and usually lasted up until 24.00 h into the night.

Amphibians and reptiles were actively searched for and

were collected by hand when encountered. Call records

were made for calling amphibians using a Marantz

recorder with an external microphone. All recorded

calls were analyzed using the software Audacity (http://

audacityteam.org) and compared against currently known

anuran call databases. Pools on the road and in other

potential locations were checked for tadpoles. Searches

for amphibians and reptiles were conducted during

the day. All collected specimens were photographed,

and afterwards were sedated and sacrificed in the field

using Lidocaine®. Specimens were preserved in 10%

formalin (one part full-strength formaldehyde and nine

parts water) and after a few days they were transferred

to 70% ethanol for storage at the National Zoological

Collection of Suriname (NZCS). The identities of some

of the amphibian specimens collected were verified by

molecular analyses, as indicated in Table 1. Additionally,

some data are provided below on the habitats where the

species were collected, and whether they are documented

here for the first time for the Lely Mountains.

Historical survey data. To complete the present list of

species known from the Lely Mountains searches were

performed in the online databases from Naturalis (http://

bioportal.naturalis.nl) and GBIF (https://www.gbif.org/

species/search) to check for species from this location that

were not formally mentioned in the published literature.

Figure 2 gives an overview of the total number of species

documented in the three herpetological surveys at the Lely

Mountains (Hoogmoed in 1974/5, Watling and Ngadino in

2006, and the fieldwork reported here in 2016).

Results

A total of 28 species of amphibians, representing

seven families, and 18 species of reptiles in 11

families, were collected (Figs. 3-8). All specimens

were unambiguously identified to the species level.

Previously, 19 anuran species were recorded for the

Lely Mountains (Watling and Ngadino 2007). The two

lists combined yield a total of 29 species, eight of which

were never before documented for the Lely Mountains.

For the Lely Mountains, the RAP (Rapid Assessment

Program) reported a total of 18 reptile species (Watling

and Ngadino 2007); the present survey also collected

a total of 18 species, seven of which were new records

for this location. Database searches of Naturalis (http://

bioportal.naturalis.nl) and GBIF (https://www.gbif.org/

species/search) provided a total of 34 anuran species, 17

of which were not documented during either the RAP or

the survey held in June 2016. For reptiles, Naturalis and

GBIF yielded a total of 25 species, 13 of which were not

documented in the surveys. Taking into account all of

the surveys and database reports for the Lely Mountains

yields a total of 46 anurans and 41 reptiles identified to

the species level (Table 1).

Some noteworthy observations were made for some of

the amphibian and reptile species collected in June 2016.

* Anomaloglossus stepheni was collected for the first

time in the Lely Mountains. Specimens were collect-

ed in the leaf litter in a patch of dry pristine forest

approximately one km north of the airstrip.

¢ Boana xerophylla is formally reported for the first time

for the Lely Mountains, and was collected during the

Table 1. Amphibian and reptile species reported for Mount Lely, Suriname. Data are included from three main sources: the Rapid

Assessment Program (“RAP 2007”), the current survey conducted in June 2016 (“Current 2016”), and all others from literature

surveys and online sources (“Other”). The latter include literature references, when available, and asterisks indicate the original

collector(s), when known. For species only reported from one of these three sources, the table cells are color coded (yellow, green,

or blue), which emphasizes the inherently “incomplete” nature of any individual survey. The last column (“Seq”) indicates whether

sequences are available.

Hisar taxa eit Current Original collector/literature

8 P se a sen __

[Amphibiamara

a ae)

[Atomobatdee | Allobaresfemoratis | | x | _X | Woogmosa 7s" | X_

Bufonidae Rhinella margaritifera species

complex

Atelopus hoogmoedi — SF Ouboter and Jairam 2012 =

Amphib. Reptile Conserv. 162 November 2019 | Volume 13 | Number 2 | e200

Rawien Jairam

Table 1 (continued). Amphibian and reptile species reported for Mount Lely, Suriname. Data are included from three main sources:

the Rapid Assessment Program (“RAP 2007”), the current survey conducted in June 2016 (“Current 2016”), and all others from

literature surveys and online sources (“Other”). The latter include literature references, when available, and asterisks indicate the

original collector(s), when known. For species only reported from one of these three sources, the table cells are color coded (yellow,

green, or blue), which emphasizes the inherently “incomplete” nature of any individual survey. The last column (“Seq’’) indicates

whether sequences are available.

igher taxa epee 2007 | 2016 citation ad

a —— | (a | Cy Cac)

a a

Caste [Beane serpinla |

isiae | Denton excoptens [|X| | Hoge 175

C ivse | Dendopsops mina [| |x| Hoopmecs 1975"

Hylidae Dendropsophus melanargyreus yi 4 Hoogmoed and Avila Pires [|

1991

tiyiase | Dendropsophas gauchers | | [MN Fougueterai 27 [| _

[ivliae | Oseocephas oophagus | | Ia

Ttivliae | Oseocephatusaurims |X | [| _X | Hoogmoed 75" | _

Tfivlidae | Osteocephats teprewt | | [RIN Oubover and Jairam 2072 |

Ttivlidae | Oseocephats tence | | [RIE oogmoed and Poder 1975" [

[Phyttomedusiae | Puhecopus pochondriats [|X | x | _Myersio7s | _

[Phyltomedusidae | Callnedusatomopnerna | |X | _X | Hoogmoea i975" | __

[Phyliomedusidee | Plyliomedisa vain | | [IN Hoogmoca 75" [__

a So =

Trias | Sein protoscidens | | MN Hoogmocao7s® |

Triylidae | Sein boesemari____| | ___[N——_Hoogmoea 975" | __

a — aaa

Leptodactylidae Adenomera heyeri —- and es 1957:

SSS et al. 2011

js i a

Tceptodaeylidae [Leprodacpiustongirstis | _x [x | | sd

Tceptodactylidae [ Leprodacyius mstaceus | x [x _| _X | Hoognoed and Poder 1975" [__

Tceptodaetylidae | Leprodacyius pentadacyus | x [x |x | Hoogmoed 1975*_ | __

TKeprodacyldae [Lepradacytus senodema | | JRE Hoozmoed ana Myers 1775" |

Tceprodacylidae [Leprodacytus guianensis | [|X |X | Hoogmoed 975 |X

Tceprodacyldae | Leprodactusrhodomystax | | x | _X_| Mooginoed and Poker 1973" |_x_

TLeprodacylidae [Leprodacytus peers | | REN)—Ouborer and Jaram 2012 |

a TY po

[Strabomantiae [Prsimanis ngunatis |__| [RIN Outer and ara 019 |

TStrabomanidae [Prisimanns marmorans |_| __[J———Myers 1975]

[Swabomantiae [Pristmanissps _|_x [x |] Ss

Amphib. Reptile Conserv. 163 November 2019 | Volume 13 | Number 2 | e200

Herpetofauna of the Lely Mountains, Suriname

Table 1 (continued). Amphibian and reptile species reported for Mount Lely, Suriname. Data are included from three main sources:

the Rapid Assessment Program (“RAP 2007”), the current survey conducted in June 2016 (“Current 2016”), and all others from

literature surveys and online sources (“Other”). The latter include literature references, when available, and asterisks indicate the

original collector(s), when known. For species only reported from one of these three sources, the table cells are color coded (yellow,

green, or blue), which emphasizes the inherently “incomplete” nature of any individual survey. The last column (“Seq”) indicates

whether sequences are available.

Hivher taxa Riteiaa Current Original collector/literature

8 P a a seen __

| Amphibia: Anura | Anura

ee! Gee

TStrbomantidee [Pristimants guturais | | __f]—Outover and Jira 2012 [__

Pe A

Amphibia:

eunnonueng

Microcaecilia grandis | Wilkinson et al. 2009 et al. 2009

Total amphibians 49 species 19 28 33

[ReptarSawria [SC SSE Crd dTTCCSCSdr

[Phyllodciyliae | Thecadachs rapid = a ae

TGekkonidee | Lepiodacttus ngubris | _ I

TGekkonidae | Hemidacyus mabouia [___ a

[sphaerodacylidae | Gonatodes hamerais _[_X [x | _X | Hoogmoes 7 [

[Gymnnoptaimidae | Loxopholisganense [x [x | _X | Hoogmoea ior _[

[Gyinoptaimidae | Nenscuras bcurinarus [| x _| _X__| Hoogioed and Polder 1975" | __

[Gymnoptaimidae [.Nensticurasruds | _X [| X | Hoogmoea v7" _[

Gymnopthalmidae Arthrosaura reticulata Hoogmoed and Avila-Pires

1992

Gymnopthalmidae Arthrosaura kocki xX xX Hoogmoed and Avila-Pires

1992

[Daciyloitae | Anotischrwoteps | x | | _x_ | Hoogmoed io75" | _

TDactyloitee | Anotispunctans | | [RI Hoogmoea 975" | _

[Potyehrotae | Pobyohras marmoranns | |

[ Seinciae | Copecglossum nigropuncranm | x |__| X | Hoogmoed 75" | __

[Gyinnophaimidae [iphisactegans |_| [RIN Hoogmoea 975° | __

TGyinnophthamidae | Treoscinensagiis | | [MN —_Hoogmoea 975" | __

TSpacrodaciylidae | Chuogetto amaconicus | | [MN] Hoogmoea 975" [| _

teidee —___[Ameivaamena | x |X | _X_ | Hoogmoea i975" | _

[reine | Papinambisegutsin | x |x [| ———s+ds

[tropiauridae | Plcaplica __—————~it x ‘|| x | GkMesm | _

[tropiduidae [Plea umbra | | | x | Hoogmeedi975* | __

Repti: Crocodyia | Ci CT TT Cd

TAllgatordae [Caiman crocodius | | [NN] —_Hoognoeaionse | __

Repti serpents | PT CSC

[ Leptotyphopidae | Stagonodon capmensts | | [IN Hoognocaio77 | __

[ceptotyphopidae [Apia enetta =i So —

Amphib. Reptile Conserv. 164 November 2019 | Volume 13 | Number 2 | e200

Rawien Jairam

Table 1 (continued). Amphibian and reptile species reported for Mount Lely, Suriname. Data are included from three main sources:

the Rapid Assessment Program (“RAP 2007”), the current survey conducted in June 2016 (“Current 2016”), and all others from

literature surveys and online sources (“Other”). The latter include literature references, when available, and asterisks indicate the

original collector(s), when known. For species only reported from one of these three sources, the table cells are color coded (yellow,

green, or blue), which emphasizes the inherently “incomplete” nature of any individual survey. The last column (“Seq”) indicates

whether sequences are available.

Thieher taxa Sanins Current Original collector/literature

8 P site a Stren _

Repti Semenes

Sa a | a ee

poids | Corals canis | | EN GEMees 97" | __

Tcoutridae | Atractus badius | | [IN Hoogmoca i975" [| __

[Colutridae | Mastigodryas Boddoer | sc

[Cowbridae | Chironus carinaus | a

Tcowbridae | Chironiusfwcs i? | Orv |

eS a a ie

[Vipers __[Bortropsarox +t x ‘|x [| | sSsSsSsSsd—sS

Vipers [Bothrops brass ————~| | i tndeman os" |

—

PReptitiasTetuaines | YC TCS

[ Cheldse | Platenpplaycephala | x | x |X | Woogoeaio7® | _

Total reptiles 42 species

June 2016 survey. All specimens observed were found ground in a habitat of undisturbed forest.

around the houses on water tanks and rain gauges at the

airstrip. Although this species was observed and possi- ¢ Pristimantis sp. 4 is noted for the first time for the

bly collected by the members of the RAP, the specimens Lely Mountains.

were misidentified as an undescribed Pristimantis spe-

cies. For example, a misidentified specimen is depicted

° Thecadactylus rapicauda was found on some of the

on the cover of the RAP report (Alonso and Mol 2007).

buildings at the airstrip, apparently having moved to

these locations from the surrounding forests.

* Osteocephalus oophagus was heard calling while

surveyors were walking at night in a patch of un-

disturbed forest south of the airfield in the Lely

Mountains.

¢ Lepidodactylus lugubris is an introduced SE Asian

gecko (Hoogmoed and Avila-Pires 2015), and was

most probably introduced into the Lely Mountains

by the frequent flights which supply the small-scale

time for the Lely Mountains. Specimens were ob-

served on the large water holding tanks found around

yn ¢ Hemidactylus mabouia represents the first record for

the houses on the airstrip.

yet another introduced species in the Lely Moun-

tains, and was frequently observed on the building

¢ A specimen of Chironius carinatus was collected where the surveyors stayed.

near one of the buildings at the airstrip on the tap of

one of the water tanks. ¢ Polychrus marmoratus was collected for the first

time for this location, near the edge of the airstrip.

*A specimen of Trachycephalus typhonius was col-

lected during a night survey while surveyors were

walking on a Suralco road. The collected individual

was sitting on a leaf approximately 1.5 m above the

¢ Two specimens of Epictia tenella were collected in

the morning around 8 AM, while the surveyors were

walking around the edges of the airstrip. Both speci-

Amphib. Reptile Conserv. 165 November 2019 | Volume 13 | Number 2 | e200

Herpetofauna of the Lely Mountains, Suriname

mens were near the grassy edges of the airstrip and

were active when observed.

° Mastigodryas boddaerti was observed whilst lifting

a corrugated plate left behind in one of the aban-

@ Amphibians

ages doned small-scale miner camps.

*The pristine forests around the airstrip contained

relatively large, water-filled depressions which were

attractive to species such as Chiasmocleis shudika-

i Hes PANEL OUEL LON rensis and Platemys platycephala.

Fig. 2. Total number of species per group collected during

each of the major surveys in the Lely Mountains, Suriname. * Roads on the plateau with large water-filled potholes

“RAP” is the Rapid Assessment Program survey (Alonso and

Mol 2007); “Jun-16” is the current survey; “Other collectors”

includes all other available data.

were an ideal habitat for Phyllomedusa bicolor, Cal-

limedusa tomopterna, Pithecopus hypochondrialis,

and other Hylidae.

aE .< ie

;

Fig. 3. (A) Anomaloglossus stepheni; (B) Allobates femoralis;, (C) Allobates granti; (D) Rhinella martyi;, (E) Boana xerophylla,

and (F) Scinax ruber.

Amphib. Reptile Conserv. 166 November 2019 | Volume 13 | Number 2 | e200

Rawien Jairam

ove . : "

Recs; ]

“ :

; “oy ei .

. .

S. 3

*N ~ ec? ‘ }

SS ; ‘, mes - — —-

‘es!

, am °

a wu a

Fig. 4, (A) Trachycephalus typhonius, (B) Leptodactylus knudseni (juv.); (C) Leptodactylus pentadactylus (juv.); (D) Leptodactyl us

mystaceus, (E) Leptodactylus rhodomystax (juv.); and (F) Ameerega trivittata.

Discussion

Some problems were encountered with two of the species

collected during the RAP, namely Anomaloglossus

beebei and Anomaloglossus degranvillei. The authors of

the RAP survey confused A. beebei with Allobates granti,

which occurs in the Lely Mountains whilst the former is

only found at Kaieteur National Park, Guyana (Kok et

al. 2006). The Anomaloglossus degranvillei reportedly

collected during the RAP might have been confused with

A. stepheni or A. surinamensis. A study by Fouquet et

al. (2018) has shown that A. degranvillei is restricted to

a small area in French Guiana and also proved that A.

surinamensis probably occurs in the Lely Mountains.

Specimens of A. stepheni were collected during the 2016

survey. Anomaloglossus surinamensis was first described

Amphib. Reptile Conserv.

by Ouboter and Jairam (2012) who had previously also

identified specimens from the nearby Nassau Mountains

as A. degranvillei (Ouboter et al. 2007).

Some of the Pristimantis species documented during

the RAP still remain to be identified, so chances are that

their eventual identification might change the number

of amphibian species for the Lely Mountains. Since the

2016 survey was held in a period with very little rainfall,

very few frogs were calling. Therefore, we believe that

additional surveys during the rainy season would be

interesting and would definitely increase the number of

known amphibians for this location.

The increasing human presence in the Lely

Mountains might have resulted in an extension of the

distribution ranges for Boana sclerophylla, Scinax ruber,

Trachycephallus typhonius, Lepidodactylus lugubris, and

November 2019 | Volume 13 | Number 2 | e200

Herpetofauna of the Lely Mountains, Suriname

Hemidactylus mabouia; species that were not recorded

during the earlier surveys. Although most of the days

spent on the Lely Mountains were quite dry not many

reptile species were collected. Snakes, for example, were

the least represented group, and just four specimens

belonging to three species were found. Small-scale gold

miners were very active in the Lely Mountains, a factor

which might have contributed to the disturbance in the

forest and the only creek that was found and sampled.

The number of species added to the list published during

the RAP and the prospects for organizing another survey

to this location validate the value of this checklist. We

would strongly recommend that the Lely Mountains

be spared from further destruction by illegal mining

activities, and additional surveys involving other groups

should be conducted to establish a satisfactory list

Amphib. Reptile Conserv.

Fig. 5. (A) Leptodactylus longirostris, (B) Boana boans; (C)

Callimedusa tomopterna, (D) Phyllomedusa bicolor, and (E)

Pithecopus hypochondrialis. Photos by D. Baeta.

of species present and to save this Mountain from the

complete destruction of its habitats.

Conclusions

The compiled results presented herein show that the

amphibian and reptile communities on the Lely Mountains

are much more diverse than previously reported. Though

five separate surveys have been conducted at this locality,

chances are that the known species diversity will continue

to increase as different areas on the plateau are surveyed.

The presence of illegal small-scale gold miners on the

plateau has resulted in the rapid conversion of forested

areas into unsuitable habitats which may translate into a

loss of species diversity.

Acknowledgements.—| would like to thank the Nature

Conservation Department that kindly provided the

permits which allowed the fieldwork in the Lely

Mountains. D. Baéta and T. Gazoni were valuable

companions in the field, and contributed significantly

to the number of species documented during the survey

in June 2016. A first draft of this article was reviewed

by A. Fouquet whose comments greatly improved the

manuscript.

November 2019 | Volume 13 | Number 2 | e200

Rawien Jairam

eet eT dies ata

RS ~ Peay ects, OF RE ee

we >: SS

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Fouquet A, Noonan BP, Blanc M, Orrico VGD. 2011.

Phylogenetic position of Dendropsophus gaucheri

(Lescure and Marty 2000) highlights the need

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relationships of Dendropsophus (Anura: Hylidae).

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Fouquet A, Orrico VGD, Ernst R, Blanc M, Martinez Q,

Amphib. Reptile Conserv.

Fig. 6. (A) Adenomera heyeri; (B) Dendropsophus leucophyllatus, (C) Chiasmocleis shudikarensis; and (D) Pristimantis sp. 4.

Vacher JP, Rodriques MT, Ouboter P, Jairam R, Ron S.

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Hylidae) of the parviceps group from the lowlands of

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Hoogmoed MS. 1973. Notes on the Herpetofauna of

Surinam IV: The Lizards and Amphisbaenians of

Surinam. Biogeographica 4. W. Junk Publishers, The

Hague, Netherlands. 419 p.

Hoogmoed MS, Avila-Pires TCS. 2015. Lepidodactylus

lugubris (Duméril and Bibron 1836) (Reptilia:

Gekkonidae), an introduced lizard new for Brazil,

with remarks on and correction of its distribution in

the New World. Zootaxa 4000(1): 90-110.

Jairam R, Jairam-Doerga S. 2016. Range extension and

some morphological characteristics of Ptychoglossus

brevifrontalis —_ Boulenger, 1912 (Squamata:

Alopoglossidae) in Suriname. Amphibian & Reptile

Conservation 10(2) [General Section]: 30-33 (e127).

Jairam R, Jairam-Doerga S. 2015. First record of

Amapasaurus tetradactylus Cunha, 1970 (Squamata:

Gymnopthalmidae) in Suriname. Check List 11: 1-4.

Jairam R, d'Orgeix CA, d'Orgeix CH, Harris A. 2016.

Range extension and distribution of the invasive

Moreau’s Tropical House Gecko, Hemidactylus

mabouia (Moreau de Jonnes, 1818) (Squamata:

Gekkonidae), in Suriname. Check List 12(5): 1-5.

Kok PJR, Sambhu H, Roopsind I, Lenglet GL, Bourne

GR. 2006. A new species of Colostethus (Anura:

Dendrobatidae) with maternal care from Kaieteur

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Herpetofauna of the Lely Mountains, Suriname

;

Says .,

4 ‘ “f

ae } m

ae! mn

Fig. 7. (A) Arthrosaura kocki; (B) Lox

umbra.

National Park, Guyana. Zootaxa 1238: 35-61.

Mol JH, Wan Tong You K, Vrede I, Flynn A, Ouboter

P, van der Lugt F. 2007. Fishes of Lely and Nassau

Mountains, Suriname. Pp. 107-118 In: A Rapid

Assessment of the Lely and Nassau Plateaus, Suriname

(with Additional Information on the Brownsberg

Plateau). RAP Bulletin of Biological Assessment

43. Editors, Alonso LE, Mol JH. Conservation

International, Center for Applied Biodiversity Science

(CABS), Arlington, Virginia, USA. 276 p.

Nielsen S, Jairam R, Ouboter P, Noonan B. 2013. A

Amphib. Reptile Conserv.

opholis guianense; (C) Neusticurus bicarinatus;, (D) Polychrus marmoratus; and (E) Plica

herpetofaunal survey of the Grensgebergte and

Kasikasima regions, Suriname. Pp. 131-144 In: 4

Rapid Biological Assessment of the Upper Palumeu

Watershed (Grensgebergte and Kasikasima) of

Southeastern Suriname. RAP Bulletin of Biological

Assessment 67. Editors, Alonso LE, Larsen TH.

Conservation International, Arlington, Virginia, USA.

176 p.

Noonan BP, Gaucher P. 2005. Phylogeography and

demography of Guianan harlequin toads (Afel/opus):

diversification within a refuge. Molecular Ecology

November 2019 | Volume 13 | Number 2 | e200

Rawien Jairam

Fig. 8. (A) Chironius fuscus; and (B) Epictia tenella.

14(10): 3,017-3,031.

Ouboter P. 2017. Amphibians and reptiles. Pp. 256-303 In:

Natural History and Ecology of Suriname. Editor, de

Din B. LM Publishers, Volendam, Netherlands. 480 p.

Ouboter PE, Jairam R, Kasanpawiro C. 2011. A rapid

assessment of the amphibians and reptiles of the

Applied Biodiversity Science (CABS), Arlington,

Virginia, USA. 276 p.

Powell R, Crombie RI, Boos HE. 1998. Hemidactylus

mabouia. Catalogue of American Amphibians and

Reptiles 674: 1-11.

Vacher JP, Kok PJ, Rodrigues MT, Lima JD, Lorenzini A,

Kwamalasamutu region (Kutari/lower Sipaliwini

Rivers), Suriname. Pp. 124-130 In: A _ Rapid

Biological Assessment of the Kwamalasamutu Region,

Southwestern Suriname. RAP Bulletin of Biological

Assessment 63. Editors, O’Shea BJ, Alonso LE,

Larsen TH. Conservation International, Arlington,

Virginia, USA. 156 p.

Ouboter PE, Jairam R. 2012. Amphibians of Suriname.

Brill Academic, Leiden, Netherlands. 376 p.

Ouboter PE, Jairam R, Wan Tong You K. 2007. Additional

records of amphibians from Nassau Mountains,

Suriname. Pp. 126-129 In: A Rapid Assessment of the

Lely and Nassau Plateaus, Suriname (with Additional

Information on the Brownsberg Plateau). RAP

Bulletin of Biological Assessment 43. Editors, Alonso

LE, Mol JH. Conservation International, Center for

Martinez Q, Fallet M, Courtois EA, Blanc M, Gaucher

P, et al. 2017. Cryptic diversity in Amazonian frogs:

integrative taxonomy of the genus Anomaloglossus

(Amphibia: Anura: Aromobatidae) reveals a unique

case of diversification within the Guiana Shield.

Molecular Phylogenetics and Evolution 112: 158-173.

Watling JI, Ngadino LF. 2007. A preliminary survey

of amphibians and reptiles on Nassau and Lely

Mountains, eastern Suriname. Pp. 119-125 In: A

Rapid Biological Assessment of the Lely and Nassau

Plateaus, Suriname (with Additional Information

on the Brownsberg Plateau). RAP Bulletin of

Biological Assessment 43. Editors, Alonso LE, Mol

JH. Conservation International, Center for Applied

Biodiversity Science (CABS), Arlington, Virginia,

USA. 276 p.

Amphib. Reptile Conserv.

Fig. 9. Overview of the airstrip in the Lely Mountains with parts of the forest visible where the June 2016 surveys were conducted.

Photo by T. Gazoni.

Rawien Jairam is an associate researcher working at the National Zoological Collection of

Suriname. Rawien has an M.Sc. in conservation biology and has been interested in the herpetofauna

of Suriname for many years. In addition to herpetology in general, he is specifically interested in

taxonomy, species descriptions, and distribution.

November 2019 | Volume 13 | Number 2 | e200

Official journal website:

amphibian-reptile-conservation.org

Amphibian & Reptile Conservation

13(2) [General Section]: 172-173 (e201).

Book Review

Islands and Snakes: Isolation and Adaptive Evolution

Bayard H. Brattstrom

Horned Lizard Ranch, P.O. Box 166, Wikieup, Arizona 85360, USA

Keywords. Behavior, biogeography, ecology, reproduction, reptiles, Serpentes, Squamata

Citation: Brattstrom BH. 2019. Book review—Islands and Snakes: Isolation and Adaptive Evolution. Amphibian & Reptile Conservation 13(2) [General

Section]: 172-173 (e201).

4.0 International (CC BY 4.0): https://creativecommons.org/licenses/by/4.0/], which permits unrestricted use, distribution, and reproduction in any

medium, provided the original author and source are credited. The official and authorized publication credit sources, which will be duly enforced, are

as follows: official journal title Amphibian & Reptile Conservation; official journal website: amphibian-reptile-conservation.org.

Received: 26 September 2019; Accepted: 26 September 2019; Published: 16 November 2019

Snakes on islands, what could make a herpetologist

happier? Islands and Snakes, edited by Harvey B.

Lillywhite and Marcio Martins, is a fun and important | hh ‘

book, with something new and fascinating in every | NSS ae BB By a, e

chapter: A tropical island with Sea Kraits coming ashore i fa aaa

to drink fresh water and to lay their eggs; a sandy Florida

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Cottonmouth Moccasins wait for falling baby birds or for

the dropped fish that the parent birds had brought to their

young. That and so much more is here in this exciting

book!

Islands are fascinating, as each has its own ecology.

Isolated and oceanic islands have depauperate faunas

due to distance and dispersal. Continental islands have

fairly normal ecosystems, though some species may be

missing and others becoming dominant. The authors of

the chapters in this book show how interesting snakes on

islands have become.

The introductory chapter by Marcio Martins and

Harvey Lillywhite discusses the geology, geography,

and evolution of islands and their snake fauna, followed

by Harold Heatwole’s chapter on the biogeography of

Sea Kraits, and then the chapter by Xavier Bonnet and

Francois Brischoux on Sea Krait behavior, distribution,

and abundance. Fun facts: Sea Kraits can pick up one-

fifth of their oxygen through their skin, they eat mostly

eels, they must go ashore on islands to lay their eggs, they Edited by

form big mating balls, and many have predatory ticks. Harvey B. Lillywhite

In Chapter 4, by Ming-Chung Tu and Harvey ad Marcio Martins

Lillywhite, we learn more about the diving responses of

Sea Kraits and the fact that after a rain, Kraits can drink

from the tiny layer of freshwater that has fallen on the Title: Islands and Snakes: Isolation and Adaptive Evolution

ocean surface...and you will also learn more about the

mating balls.

In Chapter 5, by Marcio Martins, Ricardo J. Sawaya, - Copyright: 2019

Selma Almeida-Santos, and Otavio A.V. Marqués, we JS BN: 978-0-19-067641-4

learn about the ecology of the Lancehead, Bothrops

Editors: Harvey B. Lillywhite and Marcio Martins

Publisher: Oxford University Press

Correspondence. bayard@hughes.net Pages: xii + 343; Price: USD $120

Amphib. Reptile Conserv. 172 November 2019 | Volume 13 | Number 2 | e201

Brattstrom

insularis, on a Brazilian Island; followed by Chapter 6

by Fabien Aubret on the elapid Tiger Snake, Notechis

scutatus, on one of the islands between Australia and

Tasmania, which is the breeding site of several species

of sea birds. With all this available food (shearwaters,

petrels, gulls, cormorants, skinks, and mice), these

snakes get to be as large as 1.5 m and become a hazard

for the scientists that are studying the birds!

In Chapter 7 by Robert Henderson we learn about the

Tree Boa, Corallus grenadensis, followed in Chapter

8 by a study of the ecology and variation in the Milos

Viper, Macrovipera schwizeri, by Goran Nilson.

Chapter 9, by Harvey Lillywhite and Coleman Sheehy

III, continues the important studies on Cottonmouth

Moccasins, Agkistrodon piscivorus, including their eating

baby birds and dropped fish on an island off the coast of

Florida, USA. Richard B. King and Kristin M. Stanford

bring us up to date in Chapter 11 on the decades-long

studies on Water Snakes, Nerodia sipedon insularum,

their ecology, and evolution on Lake Erie islands between

Amphib. Reptile Conserv.

173

the USA and Canada, showing both positive and negative

human impacts on the snakes.

On Catalina Island in the Gulf of California, there

is a rattlesnake that has no rattle, Crotalus catalinensis,

and Chapter 10 by Gustavo Arnaud and Marcio Martins

covers this snake’s behavior, ecology, and conservation;

and suggests that a native Night Snake, Hypsiglena

catalinae, might be a color mimic of this rattle-less

rattlesnake. Chapter 12, by Akira Mori, H. Ota, and

Koichi Hirate discusses the impact of snakes eating

baby sea turtles that are on their way from the nest to the

sea. Since there are “islands” of habitat (deserts, ponds,

areas between lava flows or between rivers), D. Bruce

Means and César Barrio-Amoroés discuss in Chapter

13 the snakes on the South American Sky Islands—the

Tepuis—with interesting results.

Bottom line: The book 1s well written by all the authors,

the pictures are for the most part quite good, and the

information is fun and exciting. BUY IT!

Bayard H. Brattstrom is Professor of Zoology, Emeritus, California State

University, Fullerton. Bayard is the author of over 300 scientific publications,

600 environmental and consulting reports, and nine books. He has been a Visiting

Professor at several Australian Universities and even studied snakes on Clarion

Island, Islas Revillagigedos, Mexico. Bayard currently lives in a solar-based

straw-bale house on top of a hill, south of Wikieup, Arizona.

November 2019 | Volume 13 | Number 2 | e201

Official journal website:

amphibian-reptile-conservation.org

Amphibian & Reptile Conservation

13(2) [General Section]: 174-180 (e202).

Possible hybridization between East Pacific Green

Chelonia mydas and Olive Ridley Lepidochelys olivacea

sea turtles in northwest Mexico

12.*Catherine E. Hart, '2°Cesar P. Ley-Quiionez, ‘F. Alberto Abreu-Grobois, °Luis Javier Plata-Rosas,

$Israel Llamas-Gonzalez, ‘Delia Karen E. Oceguera-Camacho, and '?7Alan A. Zavala-Norzagaray

'Grupo Tortuguero de las Californias A.C., La Paz, Baja California Sur, MEXICO *Investigacion, Capacitacion y Soluciones Ambientales y Sociales

A.C. (ICSAS), Tepic, Nayarit, MEXICO +Instituto Politécnico Nacional, CIIDIR Unidad Sinaloa, Guasave, Sinaloa, MEXICO *Laboratorio de

Genética y Banco de Informacion sobre Tortugas Marinas (BITMAR) Unidad Académica Mazatlan Instituto de Ciencias del Mar y Limnologia

UNAM, Mazatlan, Sinaloa, MEXICO *Centro Universitario de la Costa, Universidad de Guadalajara, Puerto Vallarta, Jalisco, MEXICO °Eco

Mayto A.C. Playa Mayto, Cabo Corrientes, Jalisco, MEXICO

Abstract.—Photographic records of sea turtle neonates and embryos which show characteristics of both East

Pacific Green Sea Turtles (Chelonia mydas) and Olive Ridley Sea Turtles (Lepidochelys olivacea) are presented.

These turtles were produced from nests laid by Olive Ridley females in the states of Nayarit, Jalisco, and Baja

California Sur. Their discovery further suggests the occurrence of hybridization between these two species,

and potential implications for conservation are discussed.

Keywords. Black sea turtle, Gulf of California, morphology, conservation, hybrid, Testudines

Citation: Hart CE, Ley-Qufonez CP, Abreu-Grobois FA, Plata-Rosas LJ, Llamas-Gonzalez |, Oceguera-Camacho DKE, Zavala-Norzagaray AA.

2019. Possible hybridization between East Pacific Green Chelonia mydas and Olive Ridley Lepidochelys olivacea sea turtles in northwest Mexico.

Amphibian & Reptile Conservation 13(2) [General Section]: 174-180 (e202).

International (CC BY 4.0): https://creativecommons.org/licenses/by/4.0/], which permits unrestricted use, distribution, and reproduction in any me-

dium, provided the original author and source are credited. The official and authorized publication credit sources, which will be duly enforced, are as

follows: official journal title Amphibian & Reptile Conservation; official journal website: amphibian-reptile-conservation.org.

Received: 24 April 2017; Accepted: 13 July 2018; Published: 19 November 2019

Introduction

Sea turtles are widely distributed throughout tropical

and subtropical oceans. They nest on sandy beaches

throughout their range, and while different species can

be found sharing habitats worldwide each has adapted to

take advantage of different ecological niches (Tomas et

al. 2001; Ballorain et al. 2010; Jones et al. 2012). Despite

this, where species of the same lineage coincide in time and

space mating and inter-specific hybridization sometimes

occurs naturally, a process which influences 25% of

plant and 10% of animal species (Arnold 1997). When

hybridization involves threatened or endangered species

it is considered to be of conservation concern (Allendorf

et al. 2001), and there is a need to determine whether

these events are a result of anthropogenic factors before

implementing management strategies (Genovart 2009).

Of the seven extant species of sea turtles, hybridization

has been reported between Loggerheads (Caretta caretta)

and Kemp’s (Lepidochelys kempii) [Barber et al. 2003];

Loggerheads and Hawksbills (Eretmochelys imbricata)

[Lara-Ruiz et al. 2006]; Greens (Chelonia mydas) and

Loggerheads (James et al. 2004); Greens and Hawksbills

(Seminoff et al. 2003; Kelez et al. 2016); and Hawksbills

and Olive Ridleys (LZ. olivacea) [Lara-Ruiz et al. 2006].

Thus, the Carettini and Chelonini tribes to which they

belong are thought to be one of the oldest vertebrate

lineages that is known to hybridize in nature (Karl et al.

1995), sharing a common ancestor more than 50 million

yr ago (Bowen et al. 1993; Dutton et al. 1996).

Along the Pacific coast of Mexico, nesting occurs

for four species of sea turtles: Leatherback, Hawksbill,

Green, and Olive Ridley, with the latter being by far the

most abundant. East Pacific Green Turtles are the second

most abundant turtle to nest on Mexico’s Pacific beaches,

and they often nest alongside Olive Ridleys. These two

species have overlapping breeding seasons with Olive

Ridleys nesting from May to December (Garcia et al.

2003) and Greens from September to January (Alvarado-

Diaz et al. 2003). Currently both species exhibit incipient

recovery and nearly year-round nesting as a result of the

population increases following decades of conservation

activities. Extensive hunting and egg collection, which

reached industrial levels in the 1970s and 1980s, led

to drastic population declines (Chassin-Noria et al.

2004; Rodriguez-Zarate et al. 2013). These declines

Correspondence. cehart03@gmail.com (*CEH), cleyg@ipn.mx (CPLQ), alberto.abreu@ola.icmyl.unam.mx (FAAG), ljplata@yahoo.com

(LJPR), israel_llamas@hotmail.com (ILG), karen@grupotortuguero.org (DKEOC), anorzaga@ipn.mx (AAZN)

Amphib. Reptile Conserv.

November 2019 | Volume 13 | Number 2 | e202

Hart et al.

Gulf of

California

20°N

Pacific Ocean

110°W

Sinaloa

MEXICO

Nayarit

Islas

Marias |

e Playa Chila

Playa El Naranjo

Playa San Francisco

@ Playa El Salado

Playa Mayto

Playa Careyeros

Jalisco

105°W

Fig. 1. Northwest Mexico. Circles denote nesting beaches where suspected hybrid hatchlings have been observed.

were reversed with a total ban on sea turtle use starting

in 1990 (Marquez et al. 1998) and the proliferation of

conservation programs. Improvements in the conditions

of these species have resulted in both being moved from

the Endangered to Vulnerable classifications on the

IUCN Red List, although both species remain listed in

Appendix I of CITES (2007).

Here, the possible hybridization of East Pacific Green

Sea Turtles and Olive Ridley Turtles is reported in

northwest Mexico based on hatchling characteristics.

Materials and Methods

During 2010-2012, hatchlings and embryos from

12 nests laid by different Olive Ridley females were

observed with characteristics (see Table 1) typically

associated with East Pacific Green Turtles. Conservation

biologists collaborating with two NGOs in north-west

Mexico, Red Tortuguera A.C. and Grupo Tortuguero de

las Californias A.C., were requested to report embryos

and neonates that were atypically pigmented, or those

that presented scute or morphological patterns associated

Amphib. Reptile Conserv.

with a species different than that of the female which had

nested. Following the subsequent reports, photographic

records of the neonates were made prior to their release.

Results

All of the hatchling sea turtles that presented atypical

characteristics hatched from nests verified to be laid

by Olive Ridley females. Figures 2A and 2B show the

contrasting morphology and coloration of the carapace

and plastron for Olive Ridley and East Pacific Green

Turtle hatchlings, respectively. Embryos (Fig. 3)

and hatchlings (Figs. 4-5) were occasionally found

presenting coloration typical of East Pacific green turtle

hatchlings. Interestingly these hatchlings were often

the only abnormal hatchlings from an otherwise typical

Olive Ridley clutch, with their siblings presenting typical

Olive Ridley coloration.

Olive Ridley hatchlings presenting a white border to

the marginal scutes and white edges to the flippers were

frequently reported (Fig. 6). We consider this to be a

normal characteristic, and not a sign of hybridization, that

November 2019 | Volume 13 | Number 2 | e202

Possible hybridization of Chelonia mydas and Lepidochelys olivacea

1 =

FS a Se Pes ee 3:

~ Ps)

Fig. 2. Carapace (A) and plastron (B) of

White

plastron Black

carapace

Fig. 3. Embryo from an Olive Ridley nest, clearly displaying East Pacific Green Turtle coloration on both (A) plastron and flippers,

and (B) carapace. Photos by C.E. Hart.

Table 1. Morphological features of putative hybrid neonate turtles compared to those usually reported for Lepidochelys olivacea

and Chelonia mydas.

Morphological feature L. olivacea C. mydas Putative hybrids

Prefrontal scales 4 2 2 or 4

Post orbital scales 3 =, 3

Marginal scutes 12 11 11-12

Supracaudal scutes 2 2 2

Intergular scute Yes

Postanal scute No

Nuchal scute Yes Yes NES

Lateral scutes 6-9* 4 4-8

Vertebral scutes 6-9 5 6-7

Inframarginal scutes

Keels Yes No Yes

Beak Triangular Rounded, large Triangular or rounded

Anterior claws 2 1 1-2

Carapace color Gray Black Dark gray or black

Plastron color Gray White White

*occasionally five

Amphib. Reptile Conserv. 176 November 2019 | Volume 13 | Number 2 | e202

Hart et al.

Keels on carapace

White border to

marginals and flippers

Fig. 4. Deceased hatchling presenting (A) white coloration to the carapace and flipper border, and (B) the white plastron characteristic

of East Pacific Green Turtles, while presenting (C) a typical Olive Ridley carapace and head. Photos by C.E. Hart.

| eC

¥ LA

‘

P aot -

Fig. 5. Healthy neonate turtles presenting characteristics of both East Pacific Green and Olive Ridley Turtles photographed before

release. Photos by C.E. Hart (A) and F- Sanchez (B).

is present in some but not all Olive Ridley neonates and Turtle neonates, and they have not been described as

is found within all participating hatcheries in southern —_ pigmentation for Olive Ridley neonates. Genetic studies

Nayarit and north Jalisco. However, a white border are needed to clarify whether these neonates are pure

on the marginal scutes and white edges to the flippers | Olive Ridleys or the result of hybridization.

are characteristics associated with East Pacific Green

Amphib. Reptile Conserv. 177 November 2019 | Volume 13 | Number 2 | e202

Possible hybridization of Chelonia mydas and Lepidochelys olivacea

Discussion

Olive Ridley turtles are the most abundant of all sea

turtle species, and this is particularly evident in the East

Pacific, with an estimated 1.39 million large juvenile and

adult Olive Ridley turtles in the Tropical East Pacific

(Eguchi et al. 2007). Because the vast differences in

abundance would greatly increase the opportunities for

hybridization of Olive Ridleys with East Pacific Green

Turtles, there is concern that if these become widespread

they could jeopardize the recovery of the much smaller

East Pacific Green Turtle population. For example,

hybrids could be sterile and so reduce overall fertility.

Hawksbill-Loggerhead hybrids have been found to be

fertile in Brazil where introgression (breeding of hybrids

with one or both parental taxa) has become significant

(Lara-Ruiz et al. 2006). Furthermore, Soares et al. (2017)

found that Loggerhead-Hawksbill hybrids were at no

reproductive disadvantage relative to the pure Hawksbills

among which they nested, suggesting that these hybrids

are likely to persist there.

Many studies report either sterility or low fitness in

hybrids (Allendorf et al. 2001). However, reports from

conservation projects in northwest Mexico suggest that

putative Olive Ridley-Green Turtle hybrid hatchlings are

Amphib. Reptile Conserv.

Fig. 6. Olive Ridley neonate from

Nayarit with the commonly found

coloration of fine white border to

carapace and fore flippers. This

coloration is not reported in the

literature for this species. Photos by

C.E. Hart.

more fit (being first to the water on release) than the Olive

Ridley turtles that hatch from the same and/or different

nests. However, the report of a more aggressive behavior

by some of these hybrid hatchlings is also worrying.

We recommend using these illustrations to aid a

national program aimed at compiling information on

the frequency and locations of occurrence of atypical

hatchlings within conservation projects, in order to

gauge the importance of this phenomenon, coupled with

genetic analyses to definitively confirm hybridization.

If confirmed hybridization is found to be abundant,

possible impacts on the regional populations are a

cause for concern. We hope this information will open

the conversation on the issue of hybridization between

sea turtle species in Mexico and highlight the need

for appropriate management guidelines to advise

conservation projects on the action to take (if any) when

these neonates occur.

Acknowledgments —CEH would like to thank the

Consejo Nacional de Ciencia y Tecnologia (CONACYT)

Mexico for support through a Ph.D. Scholarship

(number 310163). We would also like to thank the many

Tortugueros who have been keeping an eye out for

“strange hatchlings.”

November 2019 | Volume 13 | Number 2 | e202

Hart et al.

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Dutton PH, Davis SK, Guerra T, Owens D. 1996.

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PH. 2007. At-sea density and abundance estimates of

the Olive Ridley Turtle Lepidochelys olivacea in the

eastern tropical Pacific. Endangered Species Research

3: 191-203.

Espinosa-Carreon TL, Valez-Holguin JE. 2007. Gulf

of California interannual chlorophyll variability.

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Garcia A, Ceballos G, Adaya R. 2003. Intensive beach

management as an improved sea turtle conservation

strategy in Mexico. Biological Conservation 111:

253-261.

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Genovart M. 2009. Natural hybridization and

conservation. Biodiversity Conservation 18: 1,435—

1,439.

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between a Green Turtle, Chelonia mydas, and

Loggerhead Turtle, Caretta caretta, and the first

record of a Green Turtle in Atlantic Canada. The

Canadian Field-Naturalist 118: 579-582.

Jones MEH, Werneburg I, Curtis N, Penrose R, O’ Higgins

P, Fagan MJ, Evans SE. 2012. The head and neck

anatomy of sea turtles (Cryptodira: Chelonioidea) and

skull shape in Testudines. PLoS One 7: e47852.

Karl SA, Bowen BW, Avise JC. 1995. Hybridization

among the ancient mariners: characterization of

marine turtle hybrids with molecular genetic assays.

Journal of Heredity 86: 262-268.

Kelez, S, Velez-Zuazo X, Pacheco AS. 2016. First record

of hybridization between Green Chelonia mydas and

Hawksbill Eretmochelys imbricata sea turtles in the

Southeast Pacific. PeerJ 4: e1712.

Lara-Ruiz P, Lopez GG, Santos FR, Soares LS. 2006.

Extensive hybridization in Hawksbill Turtles

(Eretmochelys imbricata) nesting in Brazil revealed

by mtDNA analyses. Conservation Genetics 7(5):

773-781.

Marquez R, Jiménez MC, Carrasco MA, Villanueva

NA. 1998. Comentarios acerca de las tendencias

poblacionales de las tortugas marinas del género

Lepidochelys sp. Después de la veda total de 1990.

Ocednides 13(1): 41-62.

Seminoff JA, Karl SA, Schwartz T, Resendiz A. 2003.

Hybridization of the Green Turtle (Che/lonia mydas)

and Hawksbill Turtle (Eretmochelys imbricata) in the

Pacific Ocean: indication of an absence of gender bias

in the directionality of crosses. Bulletin of Marine

Science 73: 643-652.

Soares LS, Bolten AB, Wayne ML, Vilaca ST,

Santos FR, dei Marcovaldi MAG, Byjorndal KA.

2017. Comparison of reproductive output of hybrid

sea turtles and parental species. Marine Biology 164:

Tomas J, Aznar FJ, Raga JA. 2001. Feeding ecology of

the Loggerhead Turtle Caretta caretta in the western

Mediterranean. Journal of Zoology 255: 525-532.

Wyneken J. 2001. The Anatomy of Sea Turtles. NOAA

Technical Memorandum NMFS-SEFSC-470. United

States Department of Commerce, Washington, DC,

USA. 172 p.

November 2019 | Volume 13 | Number 2 | e202

Possible hybridization of Chelonia mydas and Lepidochelys olivacea

Amphib. Reptile Conserv.

Catherine E. Hart is a marine biologist currently completing her Ph.D. at the Centro Universitario de

la Costa, Universidad de Guadalajara, Mexico. Catherine received her B.S. from Instituto Tecnoloégico

de Bahia de Banderas, and her M.Sc. in Conservation and Biodiversity from the University of Exeter

(United Kingdom). She currently coordinates the Red Tortuguera A.C. and is an adviser on sea turtle

nesting for Grupo Tortuguero de las Californias A.C. Catherine’s current interests include the effects of

temperature and incubation techniques on sea turtle embryos and neonates, and the role that community

groups play in the conservation of sea turtles in northwest Mexico. Photo by C.P. Ley-Quifionez.

César P. Ley-Quifiénez is a Research Scientist and Technician at the National Polytechnic Institute

CIIDIR-Sinaloa, Mexico. César’s research focuses on ecotoxicology and wildlife conservation. He

holds a Ph.D. degree from Universidad Autonoma de Sinaloa, Mexico. Photo by C.P. Ley-Quifionez.

Alberto Abreu-Grobois is a Research Scientist at the Instituto de Ciencias del Mar y Limnologia

(ICML), Universidad Nacional Autonoma de México. Alberto received his Ph.D. in Population

Genetics from University College Swansea, Wales, United Kingdom, in 1983. Since then he has worked

at the Mazatlan Research Station of the ICML where he heads the Genetics Laboratory. Alberto’s

research focuses on the conservation genetics of sea turtle populations and on the temperature effects on

morphometrics and fitness of sea turtle hatchlings. Photo by Raquel Brisefio.

Luis Javier Plata-Rosas has a Ph.D. in Coastal Oceanography and is a lecturer at the University of

Guadalajara, Mexico. Luis has authored more than 600 popular science articles and several books,

including: The Physics of the Coyote and the Road Runner (2016), Myths of the XXI Century:

Charlatans, Gurus and Pseudoscience (2013), Myths of Science: Millions of People Can be Wrong

(2012), Butterflies in the Brain: Forty Flutters about Science (2006), A Scientist at the Museum of

Modern Art (2011), and The Ugly Duckling Theorem (2013). In 2014, Luis was awarded the Jalisco

State Prize for Science, Technology, and Innovation, in the category of popular science writing.

Israel Llamas-Gonzalez studied Biology at Centro Universitario de Ciencias Biol6gico Agropecuarias,

Universidad de Guadalajara, Mexico, and took a course in Zoology at the Universidad de Costa Rica

in 2003. In 2005, Israel co-founded the sea turtle conservation project in Mayto, Jalisco, Mexico,

and in 2009 he founded the NGO Eco Mayto A.C. Currently, Israel is the Director of the Sea Turtle

Conservation Program in Mayto, and collaborates with the Environmental Ministry of Panama in

studying the Hawksbill Sea Turtle population in Coiba National Park, Panama. Photo by Alexander

Gaos.

Delia Karen E. Oceguera-Camacho graduated with a Marine Biology degree in 2002, and a Master’s

degree in 2004, from Universidad Autonoma de Baja California Sur (UABCS), Mexico. Karen worked

on a sea turtle research project with the Brazilian NGO TAMAR in the states of Bahia, Sergipe, and

Espiritu Santo, from January to April 2006, where she participated in the training of sea turtle monitoring

techniques, environmental education, and community integration. Karen founded and currently heads

the sea turtle protection and nesting conservation project in San Juan de Los Planes, Baja California Sur,

Mexico, which is celebrating its 14" year of conservation efforts. She is presently focusing on projects

which offer activities that improve the social and environmental aspects within local communities

through involving local community shareholders. In 2009, Karen joined the Mexican NGO Grupo

Tortuguero de las Californias A.C., where she coordinated multiple sea turtle nesting sites until 2013

when she transitioned into the role of Executive Director. Photo by K. Oceguera.

Alan Zavala-Norzagaray is a biologist who received his Ph.D. from the Universidad de Autonoma

de Sinaloa, Mexico. Alan received his B.S. from Universidad de Occidente (Mexico) and his M.S. in

Natural Resources and Environmental from the National Polytechnic Institute CIIDIR-Sinaloa, Mexico,

where he is currently a Research Professor, Coordinator of the Wildlife Program, and an adviser on

sea turtle foraging areas for Grupo Tortuguero de las Californias A.C. His current interests include the

ecology and conservation medicine of sea turtles in foraging areas, and the role that community groups

play in the conservation of sea turtles in northwest Mexico. Photo by A.A. Zavala-Norzagaray.

180 November 2019 | Volume 13 | Number 2 | e202

Official journal website:

amphibian-reptile-conservation.org

Amphibian & Reptile Conservation

13(2) [General Section]: 181-202 (e204).

Diversity and conservation of terrestrial, freshwater, and

marine reptiles and amphibians in Saudi Arabia

‘Abdulhadi A. Aloufi, 7Zuhair S. Amr, 7Mohammad A. Abu Baker, and *Nashat Hamidan

‘Department of Biology, Taibah University, Al-Madinah Al-Munawwarah, KINGDOM OF SAUDI ARABIA *Department of Biology, Jordan

University of Science and Technology, Irbid, JORDAN *Department of Biology, The University of Jordan, Amman, JORDAN *Royal Society for the

Conservation of Nature, Amman, JORDAN

Abstract.—This review describes the diversity of the freshwater, marine, and terrestrial herpetofauna of the

Kingdom of Saudi Arabia that consists of 128 extant species and subspecies; 121 species and subspecies of

reptiles and seven species of amphibians according to current taxonomic systems. Four main categories of

threats affecting amphibians and reptiles were identified as habitat loss and degradation, water issues, human

disturbance and related activities, and legislation and public awareness; and supportive examples for each

category are provided. Key species that require urgent protection are: Chalcides levitoni, Platyceps insulanus,

Dasypeltis scabra, Hemidactylus alfarraji, Hemidactylus asirensis, Hemidactylus mindiae, Lytorhynchus

gasperetti, Pelomedusa barbata, Phoenicolacerta kulzeri ssp., Tropiocolotes wolfgangboehmei, and Varanus

yemenensis, due to their limited distribution, as well as Uromastyx aegyptia due to over-harvesting and trade.

According to the IUCN Red List, eight of these species are Data Deficient, four are Vulnerable, one Critically

Endangered, and one Near Threatened. The status of herpetofauna in Saudi Arabia is still far from being

completely understood. Nevertheless, the lack of formal conservation measures and low public concern makes

amphibians and reptiles extremely vulnerable in the near future.

Keywords. Anura, endemic, Sauria, Squamata, Testudines, threats

Citation: AloufiAA, Amr ZS, Abu Baker MA, Hamidan N. 2019. Diversity and conservation of terrestrial, freshwater, and marine reptiles and amphibians

in Saudi Arabia. Amphibian & Reptile Conservation 13(2) [General Section]: 181-202 (e204).

4.0 International (CC BY 4.0): https://creativecommons.org/licenses/by/4.0/], which permits unrestricted use, distribution, and reproduction in any

medium, provided the original author and source are credited. The official and authorized publication credit sources, which will be duly enforced, are

as follows: official journal title Amphibian & Reptile Conservation; official journal website: amphibian-reptile-conservation.org.

Received: 1 February 2019; Accepted: 8 May 2019; Published: 6 December 2019

Introduction

Over the past century, several major publications on the

herpetofauna of Saudi Arabia have appeared (Schmidt

1941; Haas 1957; Haas and Werner 1969; Al-Wailly

and Al-Uthman 1971; Farag and Banaja 1980; Hillenius

and Gasperetti 1984; Balletto et al. 1985; Arnold 1986;

Al-Sadoon 1988, 1989; Gasperetti 1988; Al-Sadoon

et al. 1991; Leviton et al. 1992; Gasperetti et al. 1993;

Schatti and Gasperetti 1994; Al-Johany 1995). Many

additional publications have provided distributional

data, taxonomic reviews, descriptions of new species,

and new records (Hussein and Darwish 2001; Wilms and

Bohme 2007; Al-Sadoon 2010; Cunningham 2010; Al-

Shammari 2012; Smid et al. 2013, 2016; Al-J ohany et al.

2014; Aloufi and Amr 2015; Alshammari and Ibrahimm

2015; Al-Sadoon et al. 2016; Alshammari et al. 2017;

Algahtani 2018; Sindaco et al. 2018).

Some of these papers presented erroneous records that

should be interpreted with caution. For example, Schatti

and Gasperetti (1994) suggested that records of Zarentola

muritanica [sic] and Tarentola annularis by Farag and

Banaja (1980) should be referred to as Hemidactylus

flaviviridis. Also, the record of Mabuya quinquetaeniata

from date gardens north of Umluy by Farag and Banaja

(1980) certainly refers to Trachylepis brevicollis.

Dekinesh (1991) included records of Mabuya vittata,

Stenodactylus petrii, Trapelus savignyi, and Sphenops

sepsoides from Faid Hema, which later proved to be

misidentifications. Some of the contributions of Masood

(2012) and Masood and Asiry (2012) to the herpetofauna

of the Asir region comprise obvious misidentifications

and erroneous records that call for amendments. For

example, Masood and Asiry (2012) reported 7° annularis,

T. mauritanica, Tropiocolotes tripolitanus, and

Trachylepis vittata from the Asir region. Considering the

known distribution ranges of the aforementioned species

(e.g., see Schleich et al. 1996), these 7arentola records

must be doubted as well. Other doubtful records include

those of Stenodactylus sthenodactylus, Trapelus pallidus,

and Trapelus mutabillis (Masood and Asiry 2012).

The conservation status of the terrestrial reptiles

Correspondence. ! aaroufi@taibahu.edu.sa, ? amrz@just.edu.jo, > Ma.Abubaker@ju.edu.jo, *nashat@rscn.org.jo

Amphib. Reptile Conserv.

December 2019 | Volume 13 | Number 2 | e204

Diversity and conservation of reptiles and amphibians in Saudi Arabia

eo aBN,

HARRAT ~4

AL HARRAH “Ay

SAKAKAH e

HAFR ALBATIN,, . ‘

AL RIYA

ALLAT NAJD PLATEAU

AL SARAWAT

MOUNTAINS

of the Arabian Peninsula was assessed by Cox et al.

(2012). Some more recent works have focused on

individual countries. Carranza et al. (2018) discussed

the diversity, distribution, and conservation of the

reptiles of Oman, including records for 101 species of

terrestrial reptiles. The distribution of reptiles in Qatar

was presented by Cogalniceanu et al. (2014). Gardner

(2013) compiled distribution data for the amphibians

and reptiles of Oman and the United Arab Emirates.

Disi et al. (2014) gave a comprehensive account of

the diversity, conservation, and major threats for the

herpetofauna of Jordan.

Since the publications of Arnold (1986) on the lizards

of Arabia, Gasperetti (1988) on the snakes of Arabia,

and Leviton et al. (1992) on herpetofauna of the Middle

East, no updated lists covering Saudi Arabia have been

published. Many reptilian and amphibian species have

been subjected to critical reviews on the molecular and

morphological levels which resulted in the adoption of

new names. In addition to documenting and updating the

herpetofauna, based on current taxonomic understanding,

Amphib. Reptile Conserv.

Fig. 1. Map of Saudi Arabia showing main geographic landmarks (after Al-Nafie 2018).

3000 m

2000

1000

SAND

SABKHAH

EDGE —

PF as

VALLEY - =“

INTERNATIONAL == = = ==

§0 55 BORDERS

it is important to examine the impacts of continuous

drastic changes in habitats and some practices in Saudi

Arabia, as the reptiles and amphibians are being subjected

to several forms of threats that have caused declines in

some species.

In this study, the diversity, conservation status, and

major threats affecting the herpetofauna of Saudi Arabia

are identified, and an updated list, including the up-to-

date taxonomic names, is provided.

Materials and Methods

The Approach

Scientific names of the reptilian species mostly follow

Uetz et al. (2018). The taxonomic treatment of the genus

Phrynocephalus by Melnikov et al. (2014) was adopted.

The systematic position of Trapelus ruderatus (formerly

Trapelus persicus) was followed after Ananjeva et

al. (2013). For amphibians, the genera listed in the

Amphibian Species of the World (http://research.amnh.

December 2019 | Volume 13 | Number 2 | e204

Aloufi et al.

Fig. 2. Ad Disah mountains, southwest Tabuk. Photo by S. Al Jathii.

org/vz/herpetology/amphibia/) were adopted. The

conservation status for species follow Cox et al. (2012).

Data on the threats affecting the reptiles and amphibians

were compiled from field observations mainly made

by the first author, A. Aloufi. Distributional limits

and localities were checked according to Sindaco and

Jereméenko (2008) for lizards, and Sindaco et al. (2013)

for snakes.

Geographical Setting

Saudi Arabia is a vast country occupying 2,026,213 km?

with diverse habitats that range from extreme arid and

basalt deserts, to mountain ranges and highlands, sand

and sandstone deserts, marine and freshwater ecosystems,

and numerous wadi systems and oases (Figs. 1-3).

Below is a general description of the geography of the

Kingdom of Saudi Arabia, including the Coastal Plains,

the Western Highlands, various plateaus, and sand dunes.

A. Coastal Plains

In Saudi Arabia, two strips of coastal plains extend along

the Red Sea and the Arabian Gulf.

The coastal plain of the Red Sea (Tihammah Plains)

lies between the Red Sea coast to the west and the western

highlands to the east. It is a narrow transitional area

between the Red Sea shelf and the high shelf mountains,

which becomes wider in the south; reaching up to 40

km wide near Jazan. To the north, it becomes narrower

until it disappears near 26°N latitude, south of Al Wajah.

With the exception of some parts near the northern tip

of the Red Sea, it 1s characterized by an abundance of

capes. The northern half of the plain is characterized

by a multitude of marine crusts, forming small marine

Amphib. Reptile Conserv.

clefts that deepen inland by water flow descending from

the coastal mountains such as Rabigh and Yanba’. The

southern half is characterized by the spread of salt flats

(Sabkha), especially along the coast, in addition to some

sandy settlements near the coast, and the spread of some

small black lava areas.

The coastal plain of the Arabian Gulf is limited to the

gulf coast to the east, and the As-Summan plateau to the

west. It is a flat plain largely covered by sand and salt flats

(Sabkha), especially near the coast and along its side.

This plain is devoid of wadi systems, with numerous sea

extensions and capes. The lowest point in Saudi Arabia

(24 m asl) lies within its southern part, near Al Homor

Sabkha.

B. Western Highlands

The western highlands consist of a mountain chain that

extends along the coast of the Gulf of Aqaba and the Red

Sea, stretching from Jordan to the north and reaching the

Republic of Yemen to the south. These mountains are a

refractive ladder-shaped formation, and its western slopes

descend precipitously towards the Red Sea, while inland

they descend gradually eastward. The altitude increases

towards the south, reaching as high as 3,015 m asl at the

mountain of Al-Sida, northwest of Abha. The western

highlands are divided into three main mountain series: Al

Sarawat mountains in the south, Al Hijaz mountains in

the middle, and Madyan mountains in the north (Fig. 2).

C. Plateaus

The hills or plateaus located to the east of the western

highlands cover large areas, and they generally descend

to the east and north-east.

December 2019 | Volume 13 | Number 2 | e204

Diversity and conservation of reptiles and amphibians in Saudi Arabia

1. Western Plateaus

Najran and Asir plateaus are located to the east of the

Sarawat mountains (Figs. 3A—B), between the Kingdom's

borders and the Republic of Yemen to the south, and the

drainage system of Wadi Al-Dawasir valley to the north. In

this area of overlap they form a transitional area between the

Sarawat Mountains in the west and the Empty Quarter in

the east. Their altitudes range between 900 and 1,700 m asl.

The Hijaz Plateau is located to the east of the Hiyaz

Mountains, and is bounded by the Hijaz mountain to the

west, the Najd Plateau to the east, the plateaus of Najran

and Asir to the south, and the Hisma Plateau to the north.

Its southern part descends to the north and west, and its

northern part descends to the east and north-east. The

altitude reaches up to 1,200 m asl. Some of its stretches

include the black lava desert of Khaybar.

Hisam Plateau is located to the north-west of the Kingdom,

to the east of the mountains of Medyan, and to the west

and north-west of Tabuk. It is confined between the border

with Jordan to the north, and Hijaz Plateau to the south.

It descends eastwards, and ranges from 800—1,700 m asl.

Black lava deserts cover some of its southern parts (Fig. 2).

2. Central Plateaus

The Central Plateau is represented by An-Nafud or Najd

plateau, a large plateau located east of the Hijaz Plateau.

It is bordered to the west by Hijaz Plateau, to the east

by the sands of Ad-Dahna, to the south by the Wadi al-

Dawasir, and by the Greater An-Nafud to the north. The

An-Nafud near Tayma lies within the Arabian shield and

is called Najd High Plateau, while the eastern part, known

as Lower Najd Plateau, lies within the Arabian shelf. It

gradually descends towards the north-east in the north, to

the east in the middle section, and to the south-east in the

south, with an average altitude of 500—1,000 m asl. The

high plateau is characterized by igneous and metamorphic

rocks. In contrast, the lower Najd Plateau is characterized

by sand and rocky edges that extend from the north near

Zulfi to the center of the southwestern extremities of the

Empty Quarter, with a length of about 1,200 km.

3. Eastern Plateaus

The Eastern Plateaus are located in the eastern part of

the Kingdom, and the extent from its center to its north-

eastern edges is all within the Arabian shelf. The Eastern

Plateaus include four main plateaus.

As-Summan Plateau is located to the east of Najd

Plateau, where it is separated from Najd by Nofud Al

Dahna, and extends from the north to the south. It is

inserted between Ad-Dahna in the west, the coastal plains

of the Arabian Gulf in the east, and from Yibreen to the

Amphib. Reptile Conserv.

south of Wadi al-Sahba in the south. This plateau is wide

and semi-flat, descending slightly towards the north-east

and east, with altitudes ranging from 50-400 m asl.

Al Hajarh Plateau is located north-east of the Kingdom,

between As-Summan Plateau in the south and Al-Hammad

Plateau in the north. It is confined between the course of

Wadi Al-Batin in the south, the Valley of Aba Al-Qor in the

north, between the Nafud Al Dahna and the Greater Dahna

in the west, and the border of the Kingdom of Saudi Arabia

with Iraq in the east. A section of Al Hajarh also extends

north of the Greater An-Nafud towards the west, reaching

Al-Jawf, and it then descends towards the east and north-

east, with altitudes ranging from 400-600 m asl.

Al Hammad Plateau is located in the north-east of

the Kingdom, and it is confined between the Al Hajarh

plateau in the south, the borders of the Kingdom with

Iraq in the north, and between Harrat Al Harrah in the

west (Fig. 3C), and Al Wedyan Plateau in the east. It

descends to the northeast with altitudes ranging from

750-850 m asl.

Al Wedyan Plateau is located in the far northeast of the

country, bordering Al Hammad plateau in the east, and

it is considered as an extension of Al Hammad Plateau,

descending towards the north-east, with altitudes ranging

from 500—750 m asl. It is crossed by several wadi systems

that descend the Al Hammad plateau and drain rainwater

to Iraq during the rainy season.

D. Sand Dunes

The sand dunes cover a large proportion of the Kingdom's

area, about 677,715 km”, or about 33.8% of its total area.

The sands of the Empty Quarter, Greater Nafud (Fig.

3E-F), and Al Jaforah are the largest sandy seas, together

representing about 90% of the sand dunes in the country.

The sand dunes are concentrated in the eastern part of

the Kingdom, while small sandy gatherings occur in

the Arabian shield to the west, in addition to small and

scattered dunes along the Red Sea coast, formed from the

presence of sediment sources, watercourses, and wind.

Biogeographical regions of Saudi Arabia

Al-Nafie (2008) defined four main phytogeographical

regions in the Kingdom of Saudi Arabia (Fig. 4). The

Saharo-Arabian region occupies the greatest part of Saudi

Arabia, extending from the north, through central Arabia.

It includes As-Summan, Al Hammad, Al Hajarh, Al

Wedyan, and Najd plateaus, An-Nafud, Ad-Dahna, and

the Empty Quarter sand dunes. The Afromontane region

has mountains higher than 1,800 m asl, is dominated

by Juniperus procera and other evergreen shrubs, and

covers a narrow strip extending along Asir and Sarawat

mountains. The Sudanian region stretches over a narrow

December 2019 | Volume 13 | Number 2 | e204

Aloufi et al.

Fig. 3. Landscapes and habitats in Saudi Arabia. (A) Juniperus procera forests in Raydah reserve. (B) Juniperus procera forests

in Asir mountains. (C) Harrat Al Harrah. (D) Harat Ewardh. (E) Sand dunes in the Greater Nofoud. (F) Sand dunes in the Empty

Quarter. (G) Elephant mountain in Al-’Ula. (H) Sharaan sand stones mountains in Al- Ula. Photos by K. Al Shamari (A), O.

Llewellyn (B), and A. Aloufi (C—H).

strip along the Red Sea coast as well as the Arabian Gulf

coast. Finally, the Sudanian-Zambian region surrounds

the Afromontane region, with overlaps with the Sudanian

region along the southwestern portions.

Results

Amphibians

Balletto et al. (1985) gave the most comprehensive

treatment to date of the amphibians of Arabia. Additional

distribution data were presented for central (Al-Johany

Amphib. Reptile Conserv.

2014) and southwestern Saudi Arabia (Al-Qahtani and

Al-Johany 2018), by Schatti and Gasperetti (1994) for

southwest Arabia, and by Alshammari and Ibnrahim

(2018) for Hai’l. Recent studies replaced Hyla savignyi

with the newly described taxon Ayla felixarabica

(Gvozik et al. 2010). Previous records of Bufotes viridis

are now considered as Bufotes boulengeri (see http://

research.amnh.org/vz/herpetology/amphibia/). In total,

seven species belonging to four families (Bufonidae,

Ranidae, Hylidae, and Dicroglossidae) are known from

the Kingdom of Saudi Arabia (Fig. 5, Table 1).

December 2019 | Volume 13 | Number 2 | e204

Diversity and conservation of reptiles and amphibians in Saudi Arabia

Table 1. List of amphibians and reptiles of Saudi Arabia, their IUCN conservation status, and levels of threats. IUCN status:

Critically Endangered (CR); Data Deficient (DD); Endangered (EN); Least Concern (LC); Near Threatened (NT); Not Evaluated

(NE); Vulnerable (VU). Threats: 1 = Deforestation, 2 = Destruction of the natural vegetation in the desert, 3 =Agricultural expansion,

4 = Overgrazing, 5 = Water extraction and climate change, 6 = Pollution and marine debris, 7 = Recreational activities and tourism,

8 = Direct persecution, 9 = Trade and commercial collection, 10 = Hunting and poaching. Levels of threats: L = Low, M = Medium,

H = High.

Tt Treats

Ampbia TCrCUdT CrP PT dT PTC TCT YC

Framity Bufonidwe Sir SCT CCT PET PT TT

[ Bufnes boulengeri Laas 187) +f we | ne | | 1 |i] |] i]

[Dunaphsmus digerensis Parker, 53) | 1c | ne |_| | |[w] | | [2[ _

[Sopris er een“ [ue [we [PT

Amphib. Reptile Conserv. 186 December 2019 | Volume 13 | Number 2 | e204

Aloufi et al.

Table 1 (continued). List of amphibians and reptiles of Saudi Arabia, their IUCN conservation status, and levels of threats. IUCN

status: Critically Endangered (CR); Data Deficient (DD); Endangered (EN); Least Concern (LC); Near Threatened (NT); Not

Evaluated (NE); Vulnerable (VU). Threats: 1 = Deforestation, 2 = Destruction of the natural vegetation in the desert, 3 = Agricultural

expansion, 4 = Overgrazing, 5 = Water extraction and climate change, 6 = Pollution and marine debris, 7 = Recreational activities

and tourism, 8 = Direct persecution, 9 = Trade and commercial collection, 10 = Hunting and poaching. Levels of threats: L = Low,

M = Medium, H = High.

a 0

tromaspmcegpia Forskal775) _|_vo | w | [a[ [al | [ep [ala

[ tromasy orata Heyden, 1827 _____| uc | 1c | |r] [xf | [et] |

FFamiyGekkonine Sid SSCS PE PT PT TT TC

[ Bunopus nbercdans Binford ira | ce | ie | | iz] |. 11.11

T Grtopodion scabram Weyden, 1827) | tc | 1c | || [z- | jefe] | _

Ee RT SD

Pepa ea es] I

Stenodactylus doriae (Blanford, 1874)

Stenodactylus grandiceps Haas, 1952

Stenodactylus slevini Haas, 1957

Stenodactylus yemenensis Arnold, 1980

Tropiocolotes nattereri Steindachner, 1901

Trigonodactylus arabicus (Haas, 1957)

Tropiocolotes wolfgangboehmei Wilms et al. 2010

ily

Acanthodactylus gongrorhynchatus Leviton and

Anderson, 1967

Family Phyllodactylidae

ERBEA Es EER RES SESE eee

ERE Be Gi eae eees sesso

BREESE BEES EBB SEE ES Eee

Pa SIPS SE) a Yl |

(cz i

if.

rd] 9 a A af a es |

Fre

—e

ims etal-2010 | DD

—

Amphib. Reptile Conserv. 187 December 2019 | Volume 13 | Number 2 | e204

Diversity and conservation of reptiles and amphibians in Saudi Arabia

Table 1 (continued). List of amphibians and reptiles of Saudi Arabia, their IUCN conservation status, and levels of threats. IUCN

status: Critically Endangered (CR); Data Deficient (DD); Endangered (EN); Least Concern (LC); Near Threatened (NT); Not

Evaluated (NE); Vulnerable (VU). Threats: 1 = Deforestation, 2 = Destruction of the natural vegetation in the desert, 3 = Agricultural

expansion, 4 = Overgrazing, 5 = Water extraction and climate change, 6 = Pollution and marine debris, 7 = Recreational activities

and tourism, 8 = Direct persecution, 9 = Trade and commercial collection, 10 = Hunting and poaching. Levels of threats: L = Low,

M = Medium, H = High.

esdina arnold Sniaooeral,2018 | ve | ne | || f1i(24[,

T wesalina adramtana (Boulenger197) | 1¢ | uc | [cfz]| |] [et 7]

[ wesalina Bernoulli (Schenkel,190)___| Ne | ne | [t[x]| || [x] |]

[ Mesalina brevrostis Banford, 1874 | 1c | uc | [rfx]| |] [2] |]

Family Varanidae

Varanus griseus (Daudin, 1803)

Varanus yemenensis Bohme et al. 1989

Atractaspis engaddensis Haas, 1950

ily

Trachylepis septemtaeniatus (Reuss, 1834)

Family Trogonophidae

Diplometopon zarudnyi Nikolsky, 1907

Family Atractaspididae

Atractaspis andersonii Boulenger, 1905

Amphib. Reptile Conserv. 188 December 2019 | Volume 13 | Number 2 | e204

Aloufi et al.

Table 1 (continued). List of amphibians and reptiles of Saudi Arabia, their IUCN conservation status, and levels of threats. IUCN

status: Critically Endangered (CR); Data Deficient (DD); Endangered (EN); Least Concern (LC); Near Threatened (NT); Not

Evaluated (NE); Vulnerable (VU). Threats: 1 = Deforestation, 2 = Destruction of the natural vegetation in the desert, 3 = Agricultural

expansion, 4 = Overgrazing, 5 = Water extraction and climate change, 6 = Pollution and marine debris, 7 = Recreational activities

and tourism, 8 = Direct persecution, 9 = Trade and commercial collection, 10 = Hunting and poaching. Levels of threats: L = Low,

M = Medium, H = High.

Platyceps saharicus Schatti and McCarthy, 2004 LC IF ecera i” kale (ietale elle Wi wile ole ell

Se ST

Calle

SITE ARES Sse Eee Seri

IME | ST eral a

itt tt tt tt et

eA De TEE TAY ale aT as

(eo

Echis coloratus Gunther, 1878

Pseudocerastes fieldi Schmidt, 1930

Reptiles

Twenty-one families in two orders, Testudines

(Cheloniidae, Dermochelyidae, Geoemydidae,

and Pelomedusidae) and Squamata (Agamidae,

Atractaspididae, Boidae, Chamaeleonidae, Colubridae,

Gekkonidae, Elapidae, Lacertidae, Leptotyphlopidae,

Phyllodactylidae, Psammophiidae, Scincidae,

Sphaerodactylidae, | Trogonophidae, Typhlopidae,

Varanidae, and Viperidae), including 55 genera and 121

Amphib. Reptile Conserv.

mien

pater

Ete

Peers

[16

Lom

incom

Lie 2

Ete

ie)

[Halternnesa morgan (Mocquard, 1905) | VU

Pie"

eros

——

—

nom

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SE SUSE Oe Eee eb Sele aa ies

AEE EEE EEE EEE

species, have been recorded from Saudi Arabia (Table 1).

Testudines

Two species of freshwater turtles belonging to two

families have been recorded from Saudi Arabia:

Pelomedusa barbata (Pelomedusidae, Petzold et al.

2014) and Mauremys caspica (Geoemydidae, Gasperetti

et al. 1993; Vamberger et al. 2013). Algqahtani (2017)

published a detailed account on the status and distribution

189 December 2019 | Volume 13 | Number 2 | e204

Diversity and conservation of reptiles and amphibians in Saudi Arabia

Fig. 4. Phytogeographical regions of Saudi Arabia (after Al-Nafie 2008).

of P. subrufa (= Pelomedusa barbata) in Saudi Arabia.

Five species of marine turtles in two families

(Cheloniidae and Dermochelyidae) are known to occur

in the Red Sea (Ross 1985; Gasperetti et al. 1993; Al-

Merghani et al. 2000; Pilcher et al. 2014; Mancini

et al. 2015). The Green Turtle, Chelonia mydas, and

the Hawksbill Turtle, Eretmochelys imbricata, are

considered as the most common species in the Red Sea

(Fig. 6), where they feed and nest (PERSGA 2004).

Squamata

Family Chamaeleonidae

Hillenius and Gasperetti (1984) listed two taxa of

chameleons in Saudi Arabia: Chamaeleo calyptratus

calcarifer Peters 1871 and Chamaeleo chamaeleon

orientalis Parker 1938. Both species are confined to the

coastal mountains of the Red Sea, reaching up to 2,500

m asl.

Family Agamidae (Fig. 7)

Fifteen species belonging to six genera (Acanthocercus,

Phrynocephalus, Pseudotrapelus, Stellagama, Trapelus,

and Uromastyx) have been reported from Saudi Arabia.

Amphib. Reptile Conserv.

The status of species of the genus Phrynocephalus were

subjected to revisions based on the morphological and

genetic differences. Melnikov et al. (2014) differentiated

between the members of the Phrynocephalus arabicus

Anderson, 1984 complex. Their study revealed that Ph.

arabicus sensu Stricto is distributed in southern Arabia

(Yemen, Oman, southern Saudi Arabia), and Ph. nejdensis

in north-western Arabia (southern Jordan, northern and

central Saudi Arabia), while Ph. macropeltis is known in

the eastern coastal Arabia (eastern Saudi Arabia, United

Arab Emirates).

Rastegar-Pouyani (2000) published a controversial

opinion on the identity of Trapelus persicus. He stated

that the specific name “ruderatus’ (Olivier, 1804)

antedates “persicus’ (Blanford 1804), so the new

taxonomic combination is Trapelus ruderatus ruderatus

(Olivier, 1804) (= the former 7! p. persicus), and the

western subspecies 7’ p. fieldi has a new taxonomic

combination. Therefore, the name “persicus” is no

longer available and comes under the synonymy of

“ruderatus.” More recently, Ananjeva et al. (2013)

clarified the systematic position of 7rapelus ruderatus

in relation to 7? persicus, and for constancy and stability

of the taxonomy of this group, the International Code of

Zoological Nomenclature ruled that these two species

are separate. In addition, Pseudotrapelus aqabensis was

December 2019 | Volume 13 | Number 2 | e204

Aloufi et al.

Ne Fe

T. Papenfuss.

Kw)

Fig. 5. Amphibians of Saudi Arabia. (A) Euphiyctis ehrenbergii. (B) Sclerophrys tihamica. Photos by

BSG

us. (B) Stellagama stellio. (C) Phrynocephalus nejdensis. (D)

Fig. 7. Agamids of Saudi Arabia. (A) Trapelus flavi

Pseudotrapelus sinaitus. Photos by A. Aloufi.

added recently to the herpetofauna of Saudi Arabia by = Cyrtopodion, Hemidactylus, _Pseudoceramodactylus,

Aloufi and Amr (2015) and Tamar et al. (2016b). Stenodactylus, Trigonodactylus, and Tropiocolotes), in-

cluding 18 species. The genus Stenodactylus was revised

Family Gekkonidae by Arnold (1980), Metallinou et al. (2012), and Nazarov

et al. (2018), and it now includes four species. Smid et al.

This family is represented by seven genera (Bunopus, (2013) revised the genus Hemidactylus. They considered

Amphib. Reptile Conserv. 191 December 2019 | Volume 13 | Number 2 | e204

Diversity and conservation of reptiles and amphibians in Saudi Arabia

Photos by A. Aloufi.

all previous records of Hemidactylus turcicus from Saudi

Arabia as Hemidactylus granosus. Recently Smid et al.

(2016) described two endemic species for Saudi Arabia:

Hemidactylus alfarraji from Najran area and Hemidacty-

lus asirensis from Asir Province. Hemidactylus mindiae

is a new addition to the geckos of Saudi Arabia (Aloufi

and Amr 2015). Therefore, the number of Hemidacty-

lus species known from Saudi Arabia is now eight. One

record of Hemidactylus sinaitus Boulenger, 1885 from

Seir Farasan Kebir should be considered with care (see

Sindaco et al. 2014). Tropiocolotes wolfgangboehmei

is known from a single locality in central Saudi Arabia

(Wilms et al. 2010).

Family Phyllodactylidae

This family is represented by one genus and one spe-

cies, Ptyodactylus hasselquistii. Recent molecular stud-

ies revealed that the genus Ptyodactylus in the Arabian

Peninsula consists of two species complexes for P. has-

selquistii, as eastern and western clades (Metallinou et

al. 2015). A new species, Ptyodactylus ananjevae, was

described from Al Mudawwarah, southern Jordan, very

close to Tabuk Province, so it is most likely to also occur

in Saudi Arabia (Nazarov et al. 2013).

Family Sphaerodactylidae

The Semaphore geckos of Saudi Arabia are represented

by five species. Their taxonomic status was discussed by

Arnold (2009). Recently, the status of Pristurus rupestris

was evaluated on a molecular basis, which showed that

P. rupestris 1s restricted to eastern Oman, while a western

clade, Pristurus sp. 1, 1s distributed from central coastal

Oman, through Yemen, Saudi Arabia, and north to

southern Jordan (Badiane et al. 2014). The northwestern

population in Saudi Arabia may be assigned as Pristurus

Amphib. Reptile Conserv.

Fig. 8. Scincids of Saudi Arabia. (A) E. urvlepis taeniolatus. (B) rachylepis brevicollis. (C) Chalcides ocellatus (D) Scincus scincus.

guweirensis, however, we still prefer to continue

referring to Pristurus sp. 1 as Pristurus rupestris until

further studies validate their separation.

Family Scincidae (Fig. 8)

Ten species of skinks have been reported from Saudi Ara-

bia, and they are represented by six genera (Ablepharus,

Chalcides, Eumeces, Eurylepis, Scincus, and Trachyl-

epis). The three species of the genus Scincus are strictly

sand-dwelling species. Chalcides levitoni is known from

only one locality in southwestern Saudi Arabia (Pasteur

1978). Panaspis wahlbergi was erroneously reported

from Saudi Arabia by Al-Jumaily (1984).

Family Lacertidae

Twenty species of lacertids occur in Saudi Arabia. They

belong to five genera (Acanthodactylus, Mesalina,

Ophisops, Philochortus, and Phoenicolacerta). Species

of the genus Acanthodactylus were extensively studied at

the molecular level (Tamar et al. 2016a), and constitute

the highest number of species, followed by species of

the genus Mesalina. Acanthodactylus tilburyi is known

only from Saudi Arabia and southern Jordan (Sindaco

and Jereméenko 2008). Among lizards of the genus

Mesalina, Sindaco et al. (2018) described Mesalina

arnoldi from southwestern Saudi Arabia and Yemen, and

Mesalina saudiarabica was described from Mahazat as-

Sayd, near Makkah (Smid et al. 2017). By now, Mesalina

brevirostris 1s known to be distributed in eastern Saudi

Arabia, while Mesalina bernoullii is known from north-

eastern Saudi Arabia (Smid et al. 2017). Al-Sadoon

et al. (2016) recorded Acanthodactylus orientalis and

Acanthodactylus robustus for the first time from Turaif

region. Phoenicolacerta kulzeri ssp. was recently

recorded from Al Konah, Tabuk (Aloufi and Amr 2015).

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Aloufi et al.

Family Trogonophidae (Fig. 9)

This family is represented by a single species in Saudi

Arabia. The Zarudny Worm Lizard, Diplometopon

zarudnyi, was reported from several localities mostly in

eastern Saudi Arabia.

Family Varanidae

Two species of the family Varanidae were reported

to occur in Saudi Arabia. Varanus griseus is widely

distributed across Saudi Arabia, while Varanus

yemenensis is confined to extreme southwestern Saudi OSEAN PAE EE

Arabia. Fig. 9. The Zarudnyi Worm Lizard, Diplometop

Photos by A. Aloufi.

ae py

on zarudnyi.

Fig. 10. Snakes of Saudi Arabia. (A) Eryx jayakari (B) Atractaspis engaddensis. (C) Echis coloratus. (D) Cerastes cerastes. (E)

Naja arabica. (F) Walterinnesia aegyptia. Photos by A. Al Salman (A-B, F), M. Al Sulimi (C), A. Aloufi (D), and M. Al Mesheni (E).

Amphib. Reptile Conserv. 193 December 2019 | Volume 13 | Number 2 | e204

Diversity and conservation of reptiles and amphibians in Saudi Arabia

Fig. 11. Snakes of Saudi Arabia. (A) Platyceps elagantissimus. (B) Telescopus dhara.

wo BS Oeeees

, vs a ss > 70% . $26

oS .

schokari. Photos by M. Al Sulimi (A), A. Al Salman (B-C), and A. Aloufi (D).

Family Leptotyphlopidae

Two species of the genus Myriopholis, Myriopholis

macrorhyncha and Myriopholis nursii, have been

reported from Saudi Arabia (Egan 2007).

Family Typhlopidae

This family is represented by a single species, /ndotyph-

lops braminus. This Asian species is widespread and has

become almost cosmopolitan in distribution. It 1s be-

lieved to have been introduced into Arabia through im-

ported plant pots (Egan 2007).

Family Atractaspididae (Fig. 10B)

Mole vipers in Saudi Arabia are exemplified by

two species: Atractaspis andersonii distributed in

southwestern Saudi Arabia, and Atractaspis engaddensis

in the northwestern and central parts of the country.

Family Boidae (Fig. 10A)

This family is represented by two species. Eryx jaculus

is known from eastern Saudi Arabia, while Eryx jayakari

is more common and widespread throughout the country.

Family Colubridae (Fig. 11)

Amphib. Reptile Conserv.

This family includes 13 species and _ subspecies

in six genera (Dasypeltis, Eirenis, Lytorhynchus,

Platyceps, Spalerosophis, and Telescopus). Three

species are confined to the extreme southwest of Saudi

Arabia (Platyceps insulanus, Dasypeltis scabra, and

Lytorhynchus gasperetti). Eirenis coronella coronella

is known from eastern Saudi Arabia, while FEirenis

coronella fennelli is known from the western part of

the country. Schatti and McCarthy (2004) confirmed

the occurrence of Platyceps saharicus in the Arabian

Peninsula. We came across a photographed specimen

of Rhynchocalamus melanocephalus from Al Konah,

near Tabuk and another one from Jabal Al Ward west of

Al-' Ula (data not shown). However, the validity of its

occurrence in Saudi Arabia requires further specimens.

Family Psammophiidae (Fig. 11D)

This family includes two species in two genera

(Psammophis and Rhagerhis). Both species are desert

adapted species with wide ranging distributions across

the Kingdom (Gasperetti 1988).

Family Elapidae (Fig. 10E and F)

Three species of terrestrial elapids belonging to two

genera (Naja and Walterinnesia) are known in Saudi

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Aloufi et al.

Arabia. Nilson and Rastegar-Pouyani (2007) considered

the eastern population of Walterinnesia as Walterinnesia

morgani and the western population as Walterinnesia

aegyptia. Naja arabica is distributed to the west of the

country.

Nine species of marine snakes belonging to the genus

Hydrophis have been reported from the waters of the

Arabian Gulf. The nomenclature of sea snakes in Table 1

follow Rezaie-Atagholipour et al. (2016).

Family Viperidae (Fig. 10C and D)

Six species in four genera (Bitis, Cerastes, Echis, and

Pseudocerastes) of vipers are known in Saudi Arabia.

Bitis arietans, Cerastes cerastes, and Echis borkini, are

confined to the southwest, while Pseudocerastes fieldi

occurs to the extreme north on the border with Jordan

within Harat Al Harah. Cerastes gasperetti is the most

common viper, especially in sandy areas.

Relict species

Pelomedusa barbata represents a relict population and is

known from southwestern Saudi Arabia (Gasperetti et al.

1993). This African species may have either migrated to

southern Arabia or originated in Arabia (Gasperetti et al.

1993; Vargas-Ramirez et al. 2016). The Caspian Turtle,

Mauremys caspica, reaches the most southeastern range

of its distribution around Al Qatif, Al Hufhuf, and Al

Ugayr (Gasperetti et al. 1993; Fritz et al. 2008), and these

populations are considered as relicts. The population

of Phoenicolacerta kulzeri ssp. from Al Konah, in the

northwest of Saudi Arabia, represents a relict in the

sandstone formation of Hisma (Aloufi and Amr 2015).

Endemic species

Seven species are strictly endemic to the Kingdom of

Saudi Arabia. 7ropiocolotes wolfgangboehmei is known

from one locality in the proximity of Ath-Thumamah,

central Saudi Arabia (Wilms et al. 2010). Chalcides

levitoni was recorded only from Khasawyah, near Jizan

(Pasteur 1978). Two snakes, Platyceps insulanus, known

only from Sarso Island, Farasan Archipelago (Mertens

1965; Masseti 2014), and Lytorhynchus gasperetti,

known from two localities in southwestern Saudi Arabia

(Leviton 1977), are considered as endemic species.

Recently, Hemidactylus alfarraji and Hemidactylus

asirensis were described from southwestern Saudi

Arabia (Smid et al. 2016), and Mesalina saudiarabica

was described from Mahazat as-Sayd (Smid et al. 2017).

Other species can be considered as endemic at the

level of the Arabian Peninsula. Four amphibians (Dut-

taphrynus dhufarensis, Euphlyctis ehrenbergii, Scleroph-

rys arabica, and Sclerophrys tihamica) are confined to

the Arabian Peninsula. Euphlyctis ehrenbergii is distrib-

uted in southwest Saudi Arabia, and the Riyadh locality

Amphib. Reptile Conserv.

may represent a release or escaped specimens from King

Saud University Campus (Al-Johany et al. 2014). Among

the reptiles, 20 species are restricted to the Arabian Pen-

insula excluding Socotra: Acanthocercus adramitanus,

Acanthocercus yemenensis, Acanthodactylus gongro-

rhynchatus, Acanthodactylus haasi, Atractaspis ander-

sonil, Chamaeleo calyptratus calcarifer, Echis borkini,

Mesalina arnoldi, Mesalina adramitana, Naja arabica,

Platyceps variabilis, Pristurus carteri, Pristurus mini-

mus, Pristurus popovi, Scincus hemprichii, Stenodacty-

lus arabicus, Stenodactylus yemenensis, Trapelus flavi-

maculatus, Trapelus jayakari, and Varanus yemenensis.

According to the distribution maps given by Sindaco

and Jereméenko (2008), 50 out of 97 lizard species

reported from the Arabian Peninsula are considered as

endemic to the Peninsula. Twenty-five species of these

reptile species are confined to the Arabian Hotspots

Areas as outlined by Mallon (2011).

Discussion

The Conservation Status of the Reptiles and

Amphibians of Saudi Arabia

Cox et al. (2012) revised the conservation status of the

reptiles of the Arabian Peninsula, excluding the marine

snakes and turtles. Among them, 101 species are listed

as Least Concern, eight are Data Deficient, four are

Vulnerable, one is Critically Endangered, and one 1s Near

Threatened (Table 1). However, some species listed as

Least Concern, such as Echis borkini, Bitis arietans, and

Naja arabica, are under threats due to human practices

and their status assessments require revision. Other

species that are listed under Data Deficient, such as

Acanthodactylus gongrorhynchatus, Chalcides levitoni,

Lytorhynchus gasperetti, Ophisops elbaensis, Platyceps

insulanus, Tropiocolotes wolfgangboehmei, and Varanus

yemenensis, have very restricted and very narrow ranges

of distribution and so they should be assigned with

conservation priorities.

Threats Affecting the Herpetofauna in Saudi Arabia

Four main categories of threats affect the amphibians and

reptiles of Saudi Arabia. Some of these threats are very

critical for a particular species, while other species may

be impacted by more than one type of threat that may

lead to population decline.

1. Habitat Loss and Degradation

The human population in Saudi Arabia has increased

more than seven-fold in the last 50 years, now reaching

up to about 33 million. This rapid growth resulted in the

expansion of cities and urban centers at the expense of basic

natural resources, especially wildlife and their habitats.

December 2019 | Volume 13 | Number 2 | e204

Diversity and conservation of reptiles and amphibians in Saudi Arabia

Fig. 12. Habitat disturbance due to farming. (A) Fodder farms at Al Jawf. (B) Fodder farm in Tabuk. (C) Vegetable farms in Tabuk

area. (D) Farmland in Al-' Ula. Photos by A. Al Rabdi (B-C), and A. Aloufi (D).

Deforestation. It is estimated that forests and woodland

cover about 27,000 km? of Saudi Arabia. These

woodlands are mostly scattered along the Sarawat

mountain in the southwestern region and are dominated

by Juniperus procera. They have been subjected

to misuse through exhaustive wood cutting and

overgrazing activities, as well as uncontrolled forest

fire and urbanization (El-Juhany 2009). Dieback of J.

procera forests in the Raydah reserve of southwestern

Saudi Arabia was attributed to climate change and the

scarcity of rainfall, in addition to extensive farming

projects around the area (Fisher 1997).

The southwestern mountains are the home of

some rare, endemic, and perhaps endangered species

such as Acanthocercus adramitanus, Acanthocercus

yemenensis, Chalcides levitoni, Chamaeleo calyptratus

calcarifer, Euphlyctis ehrenbergii, — Lytorhynchus

gasperetti, and P. barbata. The conservation status

of these species should be assessed in relation to the

current state of vegetation cover.

Destruction of the natural vegetation in the desert.

As a result of agricultural expansion in the sand deserts

and wadi beds, the clearing of natural vegetation such

as Acacia raddiana, Acacia tortilis, Moringa peregrine,

Nitraria retusa, Ziziphus spina-christi, Retama reaetam,

and Haloxylon persicum, and other chenopods has

occurred. This clearance had direct effects on the lizards

that either use these plants for shading or feed on them.

Some species, such as TJrapelus persicus, take refuge

among Nitraria retusa shrubs. Uromastyx aegyptia is an

herbivorous lizard that feeds on a variety of desert plants

such as Pennisetum divisum and Stipagrostis plumose,

while Haloxylon salicornicum, Polygala_ erioptera,

Amphib. Reptile Conserv.

and Aerva javanica are consumed to a lesser extent

(Cunningham 2000).

Agricultural expansion. Over the past 30 years, large

parts of the deserts of Saudi Arabia were dedicated to

agricultural projects for the production of wheat, and were

converted recently for the production of green fodder and

other crops (Fig. 12). Most of these projects are located

around Tabuk, Tubarjal, Al Jawf, Hail, Buraydah, Sar,

Al Ula, and Wadi Al Dwasir, and were established since

the early 1980s. It is estimated that an area of 694,549

ha was cultivated with fodder, wheat, and other crops

in 2013. These projects were established in sandy areas

and around wadi courses, due to the availability of

groundwater. Alshammari and Ibrahim (2015) found

that the smallest numbers of reptiles were collected from

cultivated areas in Faid Hema, Ha'il region.

Much destruction of the natural habitats of sand-

dwelling species takes place due to the plowing and

construction of secondary roads in the desert. The

most heavily affected species include Acanthodactylus

schmidti, Cerastes gasperetti, Diplometopon zarudnyi,

Eryx jaculus, Lytorhynchus diadema, Phrynocephalus

nejdensis, Scincus hemprichii, Scincus mitranus, Scincus

scincus, Stenodactylus doriae, Varanus griseus, and

Uromastyx aegyptia.

Overgrazing. The nomadic lifestyle is still practiced in

many parts of Saudi Arabia. Sheep, goats, and camels are

among the domestic animals roaming the deserts and the

mountains during spring for grazing. This affects reptiles

in many ways, including direct disturbance, lowering

the vegetation cover, and actually abolishing entire plant

communities. Species such as Trapelus persicus are af-

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Aloufi et al.

fected by grazing and are displaced from their natural

habitats due to the year-round camel grazing on Nitraria

retusa shrubs.

2. Water Issues

Water extraction and climate change. Intensive farm-

ing in many parts of the Saudi deserts for the production

of wheat and fodder over the past 30 years has caused

drastic changes in the water table. This has led to a dra-

matic decrease in natural vegetation cover and drying of

natural ponds in the desert. The annual rainfall has shown

a significant decrease (47.8 mm per decade), with a rela-

tively high inter-annual variability, while temperatures

(maximum, mean, and minimum) have increased sig-

nificantly at rates of 0.71, 0.60, and 0.48 °C, respectively

(Almazroui et al. 2012). Eventually, the water re-charge

of aquifers will be affected for years to come. Accord-

ing to Alqahtani (2017), Pelomedusa barbata is facing

threats in southwestern Saudi Arabia as a result of rainfall

scarcity in recent years. In Al Hasa region, the number of

breeding sites of Mauremys caspica was reduced from

159 in the early 1970s to about 19 in 2009, due to de-

struction of natural springs and construction of cemented

canals, along with agricultural expansion (Aloufi 2009).

Pollution and marine debris. Cement dust pollution

has affected the Green Turtle, Chelonia mydas, in Ras

Baridi, reducing hatchling emergence success to only

40% due to the formation of hard domes above the nests

which prevent emergence and cause mortality (Pilcher

1999). As a result of urban sewage discharge in open

waters of the Red Sea, algal bloom formations have been

associated with fibropapillomatosis disease, which is

considered deadly to sea turtles (PERSGA/GEF 2004).

Marine pollution is also widely believed to affect other

sea turtles and snakes.

3. Human Disturbance and Related Activities

Recreational activities and tourism. Camping and

driving in the open areas are on the increase in Saudi

Arabia. Large vehicles and desert dirt bikes are widely

used for racing on the sand dunes. This will certainly

affect many sand-dwelling lizard and snake species, and

cause disturbance (Table 1). At the same time, many

shrubs and plants are destroyed during vehicle movement

across the terrain. Tourism causes disturbance to fragile

sensitive ecosystems, especially in places where relict or

endangered species exist.

Direct persecution. Reptiles in general, and snakes in

particular, are widely disliked animals. Many photos can

be found posted on social media which show snakes that

have been killed. Among them are venomous species,

Atractaspis andersonii, Atractaspis engaddensis,

Cerastes gasperetti, Echis coloratus, Naja arabica, and

Amphib. Reptile Conserv.

Walterinnesia aegyptia. However, non-venomous snakes,

such as Spalerosophis diadema cliffordii, Psammophis

schokari, and Platyceps rhodarchis, are also often

killed instantly even when they are encountered in the

wilderness. In Saudi Arabia, all forms of geckos are killed

since it is widely believed that they transmit leprosy.

Trade and commercial collection. We observed five

species of snakes that were traded in the animal markets

in Jeddah, Tabuk, Taif, and Al Madinah Al Monawarh:

Eryx jaculus, Naja arabica, Psammophis_ schokari,

Platyceps rhodorachis, and Spalerosophis diadema

cliffordii. These snakes were cramped in plastic bottles

and directly exposed to the sun. Lizards that were traded

in the animal markets included Chamaeleo chamaeleon,

Chamaeleo_ calyptratus, Scincus mitranus, Scincus

scincus, Uromastyx aegyptia, Uromastyx ornatus,

Varanus yemenensis, and Varanus griseus.

The Hawksbill Turtle, Evetmochelys imbricata, 1s

collected from the Red Sea and sold in animal markets

in Tabuk (Aloufi and Eid 2014). A less common item of

trade is the Western Caspian Turtle, Mauremys caspica,

but it can also be found in animal markets and pet shops

(Aloufi and Eid 2014) and sold in Riyadh and Al Hasa

Province. The population density of Pelomedusa barbata

is under stress and has decreased greatly over the last

few years in southwestern Saudi Arabia. However, it

is commonly taken from its freshwater habitats in the

southwest for trade (Alqahtani 2017).

The Spiny-tailed Lizard is sold for its meat, while

skinks (Scincus mitranus and Scincus scincus) are sold

as dried preparations that are prescribed in folk medicine

as aphrodisiacs. For animal exhibits and shows, snakes

such as Eryx jaculus, Naja arabica, and Psammophis

schokari are in demand. Most of these animals are sim-

ply disposed of after the shows, and it has been estimated

that over 100 Arabian cobras were killed in one season.

For teaching purposes, university students and high

schools purchase various amphibians, desert monitors,

and spiny-tailed lizards.

Hunting and pouching. The consumption of eggs

and meat of marine turtles has been reported in Saudi

Arabia (Miller 1989), however, this practice is not very

common. However, the major problem of hunting and

direct harvesting of the Egyptian Spiny-tailed Lizard,

Uromastyx aegyptia, is alarming. Thousands of these

animals are captured for human consumption and trade.

The locals relish both the meat and the eggs. Truckloads

of dead and slaughtered dabbs are posted on the hunter

internet sites as a sign of pride (Fig. 13). Gravid females

are in high demand due to their eggs. We counted over 17

females killed in one hunting trip, and females typically

lay 17-41 eggs in one clutch during May or June

(Bouskila 1984). This excessive harvest will certainly

affect the population of U. aegyptia, whereas large

numbers of females are killed in the eggs that are not

December 2019 | Volume 13 | Number 2 | e204

Diversity and conservation of reptiles and amphibians in Saudi Arabia

Fig. 13. The Egyptian Spiny-t

meat and eggs.

allowed to hatch. Both vitellogenic and oviductal eggs

were observed among the females that had been killed

(Fig. 13).

4. Legislative and Public Awareness

Enforcement. Although all wild animals in Saudi Arabia

are protected by law, enforcement is still far behind in

protecting the animals, and particularly reptiles. In fact,

the Spiny-tailed Lizard, U. aegyptia, is included among

the animals that are allowed to be hunted, together with

birds. The hunting season for the Spiny-tailed Lizard

is open from the beginning of August to the end of

September outside of the protected areas, with no bag

limit. This ambiguity in the number of allowed animals

has led to a massive scale of hunting and killing of

this vulnerable species. Until recently, the concept of

conserving diversity has remained obscure to many

decision makers in Saudi Arabia. The broad spectrum

of biological diversity in the country requires trained

individuals to reveal its importance for the country with

Amphib. Reptile Conserv.

ailed Lizard, Uromastyx aegyptia, 1s hunted and killed by the hundreds and sold either alive or for its

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ats

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respect to various aspects, including socio-economic,

ecotourism, scenic, and ethical perspectives.

Public awareness. As stated earlier, reptiles are disliked

by most of the locals, and surrounded by mystery and

superstitions. The general public attitude towards reptiles

is a mix of aversion and fear that stems from deep-rooted

traditional culture. Therefore, public awareness remains

one of the most important issues for introducing the

importance of reptiles to the general public; and public

awareness of the importance of conservation is a high

priority issue. The role of non-governmental organizations

is to introduce wildlife conservation to various sectors of

the community, mainly children and youngsters, as well

as to rally support from decision makers.

The environmental public awareness towards animals

in the Kingdom of Saudi Arabia remains limited, and

is almost totally lacking for reptiles. The availability of

books for providing scientific information to the public

or at the level of high school education remains very

limited. The lack of understanding in the general public is

clearly illustrated by social media being so full of videos

December 2019 | Volume 13 | Number 2 | e204

Aloufi et al.

and images of animal persecution and overhunting; with

expression of great pride in such actions.

Acknowledgments. We wish to thank Theodore J.

Papenfuss (Museum of Vertebrate Zoology, University

of California, USA), Othman Llewellyn and Khalaf Al

Shamari (Saudi Wildlife Authority), Ahmed Al Mansi

(Ministry of Environment, Water and Agriculture, Saudi

Arabia), Mohammed AI Mesheni (Oman), Abdusslam Al

Salman, Mushrief Al Sulimi, Abdulah. Al Rabadi, and

Saud Al Jathli (Saudi Arabia) for providing images for

some reptiles and the landscapes.

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Abdulhadi A. Aloufi is an Assistant Professor of Ecology, Faculty of Science, Taibah

University. Abdulhadi received his Ph.D. from King Saud University, Saudi Arabia, and his

(UAE), and Qatar.

Amphib. Reptile Conserv.

main interests are biodiversity and conservation. He has published about 20 articles and two

books on the biodiversity of the North Western region of Saudi Arabia and seabirds in Tabuk.

Abdulhadi has also written five books for children about animal welfare published by The

International Fund for Animal Welfare (IFAW).

Zuhair Amr is a Professor of Zoology and Animal Ecology in the Department of Biology,

Jordan University of Science and Technology, Jordan. Zuhair received his Ph.D. in Zoology

from the University of Rhode Island, Kingston, Rhode Island, USA. He has published over

150 papers and 10 books on various aspects of the ecology and systematics of mammals and

reptiles in Jordan, Lebanon, Syria, the Palestinian Territories, and Saudi Arabia. Zuhair serves

as the scientific authority for CITES in Jordan.

Mohammad Abu Baker is an Assistant Professor of Vertebrate Biology at The University of

Jordan. Mohammad received his Ph.D. in Biology from The University of Illinois, Chicago,

where he studied habitat selection and coexistence of African small mammals. He seeks to

understand local and regional factors that influence biodiversity and species distribution using

several empirical tools such as mark-recapture, camera trapping, foraging at experimental food

patches, wildlife tracking, and GIS. Thus far, Mohammad has over 40 publications on the

ecology, behavior, and natural history of mammals and reptiles in Jordan, Qatar, South Africa,

Egypt, and the USA. (Photo by Bruce Patterson).

Nashat Hamidan is the director of the Conservation and Monitoring Center at the Royal

Society for the Conservation of Nature, Jordan. Nashat received his Ph.D. from the University

of Bournemouth, United Kingdom. He has extensive experience in nature conservation and

reserves, and he has served in projects in Jordan, Saudi Arabia, Oman, United Arab Emirates

December 2019 | Volume 13 | Number 2 | e204

Official journal website:

amphibian-reptile-conservation.org

Amphibian & Reptile Conservation

13(2) [General Section]: 203-211 (e205).

Phylogenetic analysis of the Common Krait (Bungarus

caeruleus) in Pakistan based on mitochondrial and nuclear

protein coding genes

‘Muhammad Rizwan Ashraf, **Asif Nadeem, 7Eric Nelson Smith, ‘Maryam Javed,

2Utpal Smart, ‘Tahir Yaqub, ‘Abu Saeed Hashmi, and 7Panupong Thammachoti

‘Institute of Biochemistry and Biotechnology, University of Veterinary and Animal Sciences, Lahore, PAKISTAN *Amphibian and Reptile Diversity

Research Center and Department of Biology, University of Texas at Arlington, Arlington, Texas 76019, USA

Abstract.—Pakistan has more than 40 species of venomous snakes. One of them, the Common Krait

(Bungarus caeruleus), is responsible for most of the reported snake bites followed by Russel’s Viper, Saw-

scaled Viper, and Black Cobra. Molecular studies not only help in correctly identifying organisms but also in

finding the phylogenetic relationships and diversity among and between them. Morphological studies can be

supplemented with confirmatory molecular data to make them more authentic and accurate. This study is the

first to characterize the genetic diversity and phylogenetic relationships of Common Kraits from Pakistan,

which will help in developing effective strategies for managing snake bites through effective antivenom

development. Tail tip biopsies of 25 Common Kraits were collected from different cities in Pakistan. The whole

DNA was extracted. Four mitochondrial (ND4, Cytochrome b, 12S rRNA, and 16S rRNA) and three nuclear

protein coding (C-mos, RAG-1, and NT3) gene fragments were amplified using specific PCR primers. The

amplified DNA was sequenced by Sanger di-deoxy sequencing. Forward and reverse sequences were cleaned

and contiged using Sequencher 5.0 software. DNA data were aligned and concatenated using MEGA 6.0 and

SequenceMatrix software, respectively. Partition Finder software was used for obtaining the best partitioning

scheme and evolutionary models. Concatenated maximum likelihood and Bayesian phylogenetic trees were

constructed using RaxML and MrBayes software. The same alignments were used to perform DNA polymorphism

analysis using DnaSP 5.0 software. A percent identity matrix was created for all sequences using the online

bioinformatics tool, MUSCLE. Homology was presented in tabular form, showing the similarity among different

species of genus Bungarus. All Bungarus species were differentiated into four groups. Common Krait (B.

caeruleus) from Pakistan showed close relationships with B. sindanus and B. ceylonicus, as one monophyletic

group. The first clade included B. candidus (Indonesia, Thailand, Vietnam, and Laos), B. multicinctus (China,

Taiwan, and Burma), and B. niger (Nepal). The second clade included B. sindanus and B. caeruleus (Pakistan),

and B. ceylonicus (Sri Lanka). The third clade included B. fasciatus (Thailand and Indonesia), while the fourth

clade included B. bungroides (China) and B. flaviceps (Malaysia and Indonesia). This study traces the diversity

and phylogenetic relationships of the Pakistani elapid, Common Krait, showing the considerable inter- and

intra-specific variations from different geographical regions of the world.

Keywords. Asia, Elapidae, PCR, polymorphism, Serpentes, venomous snakes

Citation: Ashraf MR, Nadeem A, Smith EN, Javed M, Smart U, Yaqub T, Hashmi AS, Thammachoti P. 2019. Phylogenetic analysis of the Common

Krait (Bungarus caeruleus) in Pakistan based on mitochondrial and nuclear protein coding genes. Amphibian & Reptile Conservation 13(2) [General

Section]: 203-211 (e205).

4.0 International (CC BY 4.0): https://creativecommons.org/licenses/by/4.0/], which permits unrestricted use, distribution, and reproduction in any

medium, provided the original author and source are credited. The official and authorized publication credit sources, which will be duly enforced, are

as follows: official journal title Amphibian & Reptile Conservation; official journal website: amphibian-reptile-conservation.org.

Received: 25 January 2019; Accepted: 24 September 2019; Published: 17 December 2019

Introduction

Snakes are legless carnivorous reptiles of suborder

Serpentes, and their lack of eyelids and external ears

distinguish them from legless lizards (Reeder et al. 2015).

Except Antarctica and some other large islands, like

Ireland, Iceland, Greenland, the Hawaiian archipelago,

Correspondence. * asifnadeem@uvas.edu.pk

Amphib. Reptile Conserv.

and New Zealand islands, snakes are found everywhere

in the world (Roland 1994). South Asia is the region

most affected with venomous snake bites. For example,

the World Health Organization reports 35,000 to 50,000

deaths annually in India (Chippaux 1998; Pyron et al.

2013), and Pakistan reports 40,000 snake bites every

year that result in 8,200 fatalities (Pyron et al. 2013).

December 2019 | Volume 13 | Number 2 | e205

Bungarus caeruleus in Pakistan

Sie Mth ee A ipl Bieareeeed,

Daéytundi

AFGHANISTAN

Kondahér

Karachi «sac!

+ \ieor

Kunduz

aaa (STINK IAN

a) So

| :

UMEDECONTIOL)

Pua

—_—

. (Tt

iis 2

Chandigarh ;

ero

Daath |

“Ll Neuw Dethi

aie

/alpur

= Algae Po cr

Fig. 1. Sample collection sites in Pakistan for Common Krait (Bungarus caeruleus).

Venomous snakes in Southeast Asia belong to families

Elapidae (cobras and kraits) and Viperidae (typical vipers

and pit vipers). A study of hospital-admitted snakebite

cases in Pakistan revealed less than 5% neurotoxic

snakebites, and the rest were viper bites (Nisar et al. 2009).

Kraits, genus Bungarus, are identified by alternating

black and white cross-bands across the body, and are found

in all South Asian countries except the Philippines. Members

of genus Bungarus are moderate to large sized elapids

distributed in Pakistan and eastward through southern Asia

to Indonesia (Smith 1943). Currently, 12 species of kraits are

recognized (Yulin et al. 2018), including three in Pakistan.

In Pakistan, Common Kraits (Bungarus caeruleus) are

reported throughout Punjab, Khyber Pakhtoonkhwa

(KPK), Azad Kashmir, Sindh, and Southern Balochistan.

Common in the Indus valley, this 1s the only species of

kraits found in Rawalpindi and Islamabad (Khan 2002a;

Oh et al. 2019). Sindhi Krait (B. sindanus) is prevalent in

Tharparkar, Bahawalnagar, and Bahawalpur. Northern

Punjab Krait (B. razai) is reported from Mianwali (Khan

2002b). Determining the relationships among the members

of Elapidae can help in understanding their distribution and

diversity. Many studies have focused on the evolutionary

relationships of kraits.

Before the widespread use of DNA sequencing,

systematics and taxonomy were used to infer phylogenies

among species in order to explain their relationships.

Now many fields in biology are using phylogenies for a

wide variety of purposes, such as examining paralogous

relationships, population histories, dynamics of pathogens

with respect to their evolution and epidemiology (Zhou

et al. 2018; Blanquart 2019), the ontogeny of body

cells during development, and the differentiation of

tumors (Kester and van Oudenaarden 2018). Variations

in nucleotide sequences can construct phylogenies for

Amphib. Reptile Conserv.

inferring relationships among the compared sequences.

The topology of phylogeny gives some estimates about

the mutation rates, time-scales of evolutionary events,

and prehistoric movement among different geographical

regions (Soroka and Burzyfski 2018). Phylogenetics

shows relationships among organisms and genes

(Friberg et al. 2019), and can give a clearer picture of

the biodiversity, biogeography, and evolution of many

characters in related groups (Pilfold et al. 2019; Grismer

and Davis 2018; Silva et al. 2019).

The use of mitochondrial DNA data for studying

animal evolution has become a powerful tool in

the last decade. Molecular biology has helped in

these mitochondrial DNA studies to give insights

into population structure, gene flow, hybridization,

biogeography, and phylogenetics (Chandrasekaran et

al. 2019; Soroka and Burzynski 2018). Evolutionary

studies give comparisons of mitochondrial genome

organization and function while molecular studies

help to improve these evolutionary studies (Ng et al.

2019). Nuclear encoded genes seem to be a strong

source of phylogenetic information. They can be

more useful for showing the divergence of those

genes whose multiple substitutions may obscure

clear phylogenetic signals.

This is the first study from Pakistan focusing on

genetic characterization, biodiversity, and molecular

phylogenetics of the Common Krait (Bungarus

caeruleus).

Materials and Methods

A collection of 25 Common Kraits (Bungarus caeruleus)

was obtained from reptile breeders in different cities in

Pakistan (see Fig. 1, Table 1). Scalation patterns and

December 2019 | Volume 13 | Number 2 | e205

Ashraf et al.

Table 1. Common Krait (Bungarus caeruleus) samples used in this study, and their location information.

Sample ID

BC-1

BC-2

BC-3

BC-4

BC-5

BC-6

BC-7

BC-8

BC-9

BC-10

BC-11

BC-12

BC-13

BC-14

BC-15

BC-16

BC-17

BC-18

BC-19

BC-20

BC-21

BC-22

BC-23

BC-24

BC-25

tail tip biopsies were obtained from each specimen.

After DNA extraction (Sambrook and Russel 2001),

the Polymerase Chain Reaction (PCR) primers of

representative mitochondrial genes (ND4, Cytochrome

b, 12S rRNA, and 16S rRNA) and nuclear genes

Locality

Jallo Park, Lahore, Punjab, Pakistan

Balochanwali, Bahawalpur, Punjab, Pakistan

Qila Ram Qaur, Hafiz Abad, Punjab, Pakistan

Yazman Housing Society, Yazman, Punjab, Pakistan

Changa Manga Forest, Kasur, Punjab, Pakistan

Lal Suhanra National Park, Bahawalpur, Punjab, Pakistan

Rahim Yar Khan Zoo, Rahim Yar Khan, Punjab, Pakistan

Chak Risalwala, Faisalabad, Punjab, Pakistan

Qila Ram Qaur, Hafizabad, Punjab, Pakistan

New City Housing Society, Jaranwala, Punjab, Pakistan

Chak 126 GB Pind Janjua, Jaranwala, Punjab, Pakistan

Rahim Yar Khan Zoo, Rahim Yar Khan, Punjab, Pakistan

Yazman Housing Scheme, Yazman, Punjab, Pakistan

Ayub National Park, Jehlam Road, Punjab, Pakistan

Tibbi Balochan, Sadiqabad, Punjab, Pakistan

Maraghzar Colony, Lahore, Punjab, Pakistan

Lahore Zoo, Punjab, Pakistan

Raza Garden Phase 1, Sargodha, Punjab, Pakistan

Pir wala, Jhang, Punjab, Pakistan

Noor Garden, Okara, Punjab, Pakistan

Chenab Park, Multan, Punjab, Pakistan

Kalarwala, Chiniot, Punjab, Pakistan

Qadir Abad Tiba, Sadiqabad, Punjab, Pakistan

Chenab Park, Gujranwala, Punjab, Pakistan

Latitude

31°34'17.29"N

29°28'56.90"N

32°4'57.12"N

29°7'3.00"N

31°4'54.19"N

29°19'1.36"N

28°24'14.30"N

31°22'4.90"N

32°4'57.12"N

31°19'16.60"N

31°21'38.48"N

28°24'14.30"N

29°6'54.39"N

33°34'19.00"N

28°16'35.01"N

31°30'8.71"N

31°33'23.78"N

32°2'51.23"N

31°1'42.61"N

30°48'48.38"N

30°4'29.90"N

31°28'26.21"N

28°16'54.33"N

30°4'29.90"N

Longitude

74°28'36.78"E

71°59'41.86"E

73°40'49.02"E

71°45'6.73"E

73°59'53.49"E

71°54'16.43"E

70°15'32.63°E

73°1'24.80"E

73°40'49.02"E

73°23'21.44"E

73°25'28.74"E

70°15'32,63"E

71°45'17.40"E

73°4'59.00"E

70°8'6.58"E

74°14'55.48"E

74°19'33.73"E

72°37'31.68"E

72°16'45.51"E

73°28'38.33"E

71°18'51.93"E

72°33'56A1"E

70°7'45.48"E

71°18'51.93"E

(C-mos, RAGI, and NT3) from previous studies were

used for the amplification of selected regions through

PCR (see Table 2). After amplification, amplicons were

sequenced bi-directionally by Big DyeTM Terminator

on an ABI 3130XL Genetic analyzer. Forward and

Table 2. Mitochondrial and nuclear protein coding gene primers for Common Krait (Bungarus caeruleus).

Sr. No

Amphib. Reptile Conserv. 205

Gene Name Primer Sequence Source

Cyt.b 5'-TGACTTGAARAACCAYCGTTG-3' Palumbi 1996

5'-TGAGAAGTTTTCYGGGTCRTT-3' Parkinson et al. 2002

16S rRNA 5’-CGCCTGTTTAYCAAAAACAT-3 Vences et al. 2005

5’-CCGGTCTGAACTCAGATCACGT-3’ Vences et al. 2005

12S rRNA 5’>GTACACTTACCTTGTTACGACTT 3’ Knight and Mindell 1993

5’ AAACTGGGATTAGATACCCCACTAT3?’ Knight and Mindell 1993

ND4 5’-CATTACTTTTACTTGGATTTGCACCA-3’ Arevalo 1994

5’-CACCTATGACTACCAAAAGCTCATGTAAGC-3’ Arevalo 1994

RAG-1 5’ AGCTGCAGYCARTAYCAYAARATGTA3’ Chiari et al. 2004

S’AACTCAGCTGCATTKCCAATRTCA3’ Chiari et al. 2004

NT3 S’ATATTTCTGGCTTTTCTCTGTGGC3’ Townsend et al. 2008

5’GCGTTTCATAAAAATATTGTTTGACC3’ Townsend et al. 2008

C-mos 5' CATGGACTGGGATCAGTTATG 3' Lawson et al. 2005

5'CCTTGGGTGTGATTTTCTCACCT 3' Lawson et al. 2005

December 2019 | Volume 13 | Number 2 | e205

Bungarus caeruleus in Pakistan

100

Bungarus_multicinetus Bm] China

100

Bungarus_moulticinetes_ Bm9204 Taiwan

100 Bungarus_candidus_UR_BT1_Thailand

100

- Bungarvs_candidus Feba_ Indonesia

4 T

3 Bungarus_candidus FMNH_235260 Laos

Bungarus_candidus Emnam Vietnam

OK)

Bungarus_ multicineis CAS 221526 Burma

100 Bungarus_niger Bniz Nepal

Bungarus_sindanus Bein] Palostan

100 Bungarus_cevlonicus_F8 135

oo B caerulevs_4 Yarman Mandi Ponjab Palastan

100

B_caeruleus_6 Baharalpur_Ponjab_Palastan

Bcaerilens_ 1] Jaramrala_Ponjab_Palastan

B caerclevs_12 Rahim Yar Khan Ponjab Palostan

Bungarus_caervlens_UK_H? Palastan

Bungarus_fasciatus_URB24 Java_Indonesia

100

Bungarus_ fasciatus BiasT Thailani

Bungarus_bungaroides KIV9SR0196 China

Bungarvs_flaviceps_ JAM0946 Whlaysia Perak

100

Bungarus_flaviceps_MNHN_Indonesia_ Sumatra

Naja_naja_8 Thatta Thatta District Sind Palastan

0.2

Fig. 2. Mitochondrial and nuclear genes (ND4, Cyt. b, COI, 12S rRNA, 16S rRNA, C-mos, RAG-1, NT3, and BDNF) based

Maximum Likelihood phylogeny for Common Krait (Bungarus caeruleus).

reverse sequences were assembled through Sequencher

5.0 software. The resulting contigs (sequences) were

given specific identities. These contigs were then aligned

with other reported sequences obtained from the NCBI

database through MEGA (v 6.0, Tamura et al. 2013)

using the ClustalW tool for further data analyses. The

nucleotide data for each gene were concatenated using

SequenceMatrix (v1.7.8, Vaidya et al. 2011) software.

The concatenated data were partitioned through

PartitionFinder (v1.1.1, Lanfear et al. 2012) to give

the best partition scheme and evolutionary models for

phylogenetic analyses.

Two types of phylogenetic analyses, 1.e., Maximum

Likelihood (ML) and Bayesian inference (BI), were

performed through RaxML (v8.0, Stamatakis 2014)

and MrBayes (v3.2, Ronquist and Huelsenbeck 2012)

software. The resulting phylogenetic trees were

visualized and saved using Figtree (v1.4.3, http://tree.

bio.ed.ac.uk/software/figtree/) software. DnaSP (v5.0,

Librado and Rozas 2009) was used for analyzing

polymorphic sites and DNA polymorphism, to determine

the variation and genetic biodiversity in Common Krait

(Bungarus caeruleus) in relation to other species of the

genus Bungarus. Percent identity matrices were also

constructed by comparing different species of every

snake genus using online tool MUSCLE (available from

the European Molecular Biology Laboratory, https://

www.ebi.ac.uk/Tools/msa/muscle/).

Results

DnaSP software was used for analyzing the polymorphism

of the mitochondrial and nuclear genes as shown in Table

3. The ribosomal RNA coding genes showed the least

Amphib. Reptile Conserv.

variation, with lower numbers of variable sites, mutations,

and parsimony informative sites. The polymorphism

data show the variations in different mitochondrial and

nuclear genes with their conservation among Common

Krait and other species of genus Bungarus. In addition,

no significant variations were found among Common

Kraits from different cities in Pakistan.

The online MUSCLE tool was used to find relationships

among Bungarus species on the basis of homology in the

mitochondrial and nuclear genes (Table 4). This table

shows the conservation patterns in mitochondrial and

nuclear protein coding genes of various Bungarus species.

Phylogenetic analysis of Common Krait (Bungarus

caeruleus) from Pakistan was conducted using

mitochondrial and nuclear protein coding genes. In

this study, Black Cobra (Naja naja) from Thatta Sindh

was used as the outgroup for constructing maximum

likelihood and Bayesian phylogenies. The best partition

scheme and evolutionary models were used to infer the

phylogenetic relationships of Common Krait in Pakistan

with other members of genus Bungarus around the

world. Concatenated Maximum likelihood and Bayesian

Inference results gave very similar phylogenies (Figs.

2-3). All Bungarus species were divided into four main

clades. The first clade included B. candidus (Indonesia,

Thailand, Vietnam, and Laos), B. multicinctus (China,

Taiwan, and Burma), and B. niger (Nepal). The second

clade included B. sindanus and B. caeruleus (Pakistan),

and B. ceylonicus (Sri Lanka). The third clade included

B. fasciatus (Thailand and Indonesia), while the fourth

clade included B. bungroides (China) and B. flaviceps

(Malaysia and Indonesia). The first and second clades

showed a sister clade relationship with strong support

(ML BS = 100, BI PP = 1). Bungarus candidus and

December 2019 | Volume 13 | Number 2 | e205

Ashraf et al.

B caervlevs 4 Yaemnan Mandi Punjab Palastan

B_caervleus_6 Baharalpur_Penjab Palastan

0.6108

B_eaervlevs_1]_ Jaramvala_Ponjab Pelastan

0.9893 “a. : !

1 B ecaerulevs_12 Rahim Yar Khan Ponjab Palastan

Bungarus_caervlevs UK_H? Palastan

Bungarus_ceylonicus_RS 155

Bungarvs_sindanus Bein] Palastan

Bungares_candidus Beba_ Indonesia

0.9995 7 : ,

Bungarus_candidus UFR_BT1 Thailand

0.8295

Bungarus_candidus_ Bmnam Vietnam

0.9997

1+ Bungarus_candidus FMONH 253260 Lane

Bungarus multicinetus Bm] China

0.900]

Bungarus_multicinetes_Bm S204 Taiwan

0.998

Bungarvs_multicincs CAS 221326 Burma

0.9902 had ae

Bungarus_nizer Bniz Nepal

Bungarvus_fasciatus_BiasT Thailand

Bungarus_fasciatus_UR B24 Javea_Indonesia

Bungarus_bungarcides RIZ9SR0186 China

0.9475

0.9649

Bungarus_flaviceps_MMNHM_Indonesia_ Sumatra

Naja_naja_§ Thatta _Thatta_District_Sind_Palastan

0.08

Bungarus_flavieeps JAMI946 Malaysia Perak

Fig. 3. Mitochondrial and nuclear genes (ND4, Cyt b, COI, 12S rRNA, 16S rRNA, C-mos, RAG-1, and NT3) Bayesian phylogeny

for Common Krait (Bungarus caeruleus).

B. multicinctus probably diverged as separate species

only recently. Bungarus candidus, B. multicinctus, and

B. niger showed highly supported sister relationships

with a complex pattern of divergence (BI PP = 1.0

ML BS = 90). In the second clade B. sindanus and B.

caeruleus have been reported from Pakistan, thus they

are sympatric species, while B. ceylonicus (from Sri

Lanka) also showed a significant difference with strong

support through Maximum likelihood and Bayesian

inference phylogenies (PP = 1.0 and BS = 100). Pyron et

al. (2012) also revealed the same relationships between

B. caeruleus, B. sindanus, and B. ceylonicus.

In addition to the molecular data obtained, examination

of the 25 B. caeruleus specimens showed varying numbers

of ventral scales (207—218), 15 rows of mid-body scales,

and average numbers of subcaudals of 41-47.

Discussion

Elapids comprise 300 of the 2,500 known species

of snakes (Leviton et al. 2018). The Southern Asian

elapids include cobras (Naja and Ophiophagus), kraits

(Bungarus), long-glanded snakes (Maticora), and Asian

coral snakes (Calliophis) [Sanz et al. 2019]. The uncertain

phylogenetics of elapids has been a major factor for the

varying numbers of identified species of elapids in the

past (Mirtschin et al. 2017).

This study aimed to characterize the genetic

biodiversity and phylogenetic relationships of Common

Krait (Bungarus caeruleus) as there is a great deal of

unpublished data on this species. Here, mitochondrial

and nuclear protein coding genes were used to construct

the phylogeny of Common Krait from Pakistan along

with some morphological characterization. One variable

character is the number of ventral scales that ranges from

207-218 among the 25 specimens in this study. Khan

(1985) wrote a note on the taxonomic status of Common

Krait and Sindh Krait (B. sindanus), and by comparing

46 specimens, Khan noted an almost similar range of

207-218 ventral scales. The B. caeruleus in this study

had 15 rows of mid-body scales which is the same as

described by Khan (1985). Bungarus caeruleus and B.

Table 3. Polymorphism in mitochondrial and nuclear protein coding genes of Common Krait (Bungarus caeruleus).

Parameters ND4 Cytochrome b

Total number of sites 619 702

Variable number of sites 302 325

Number of mutations 302 202

Singleton variable sites 43 51

Parsimony informative sites 196 151

Segregating sites 159 151

Synonymous changes 176 156

Number of haplotypes 17 16

Haplotype diversity 0.866 0.615

Nucleotide diversity 0.08243 0.09528

Amphib. Reptile Conserv.

128 rRNA 16S rRNA C-mos RAG-1 NT3

650 520 586 802 425

270 19] 419 665 325

63 47 12 20 32

20 26 09 19 03

43 21 03 01 28

00 00 12 19 31

00 00 06 03 23

08 10 04 03 05

0.686 0.521 0.333 0.145 0.754

0.03617 0.03451 0.00288 0.00226 0.03389

207 December 2019 | Volume 13 | Number 2 | e205

Bungarus caeruleus in Pakistan

Table 4. Percent homology of mitochondrial and nuclear genes for Common Krait (Bungarus caeruleus) from Pakistan among

various Bungarus species and countries, sorted by decreasing Cytb homology. The asterisk (*) indicates the sequence from this

study, all others are based on GenBank sequences.

Homology percentages of nine representative genes

Species Country Cytb ND4 128

B. caeruleus* Pakistan 100 100 100

B. caeruleus Pakistan 99.84 99.51 100

B. ceylonicus NA 89.21 87.46

B. candidus Indonesia 89.19 84.53

B. niger Nepal 86.31 84.36

B. sindanus Pakistan 86.31 85.5

B. candidus Thailand 85.35 83.88

B. multicinctus Burma 85.19 84.34

B. multicinctus China 85.19 85.18

B. candidus Vietnam 85.19 85.67

B. multicinctus Taiwan 85.02 85.5

B. fasciatus Thailand 83.74 83.55

B. fasciatus Indonesia 83.57 83.39

sindanus had 15 and 17 rows of mid-body scales, with

the central larger row being hexagonal and white in color.

Boulenger (1897) also observed 15 rows of mid-body

scales in Indo-Pakistan Common Krait while 17 mid-

body scale rows were reported in B. sindanus from Indus

Basin. The average number of sub-caudals observed here

is 41-47 which is within the range observed by Khan

(1985): 40-54 in males and 30—54 in females. Boulenger

(1897) reported small eyes with round pupil which is

similar to those observed in this study.

There are also reports about the distribution of B.

caeruleus in Indo-Pakistan subcontinent. Eastward

it is found in Assam and Bengal (Jamal et al. 2018;

Ganesh and Vogel 2018); westward to the Pakistan-

Iran Border; Shockley (1949) and Kral (1969) reported

it in Afghanistan; Smith (1943) reported it southward

in Peninsular India and the Andaman Islands; and de

Silva (1981) also reported it in Sri Lanka. This study is

one attempt to infer the phylogenetics of B. caeruleus

in Pakistan, but suggests more studies from the above-

mentioned parts of the B. caeruleus distribution are

needed, as there are almost no studies from other parts of

of the Indo-Pakistan subcontinent on the phylogenetics

of the Common Krait B. caeruleus.

Determining the relationships among the members

of Elapidae can help in understanding the distribution

and diversity of elapids. Many studies have focused

on evolutionary relationships of elapids, and this study

examined the molecular phylogenetics of B. caeruleus

from Pakistan. The second clade (Figs. 2—3) includes B.

sindanus and B. caeruleus (Pakistan), and B. ceylonicus

(Sri Lanka). Bungarus sindanus and B. caeruleus are

sympatric species, while B. ceylonicus also showed a

significant difference with strong support through the

Maximum likelihood and Bayesian inference phylogenies

Amphib. Reptile Conserv.

93.68

90.57

i Li 2)

90.79

91.38

16S COI C-mos RAG-1 NT3 BDNF

100 100

100 99.99

96.62 100 99.4

95.08

85.65

90.09 86.11

91.91

95.5 85.03 99.38 99 62 96.63 99.76

(PP= 1.0 and BS = 100). Pyron et al. (2012) also revealed

the same relationships between B. caeruleus, B. sindanus,

and B. ceylonicus. They presented a large-scale phylogeny

of squamate reptiles for future comparative studies, and

a revised classification of squamates at the family and

subfamily levels so that taxonomy might be brought

in a line with data from the new phylogenetic studies.

Their phylogeny shows the same relationship (ML BS =

100, BI = 1) between B. caeruleus, B. sindanus, and B.

ceylonicus as is shown in this study through Maximum

Likelihood and Bayesian phylogenies.

Conclusions

Most of the currently recognized krait species

(genus Bungarus) are poorly understood. This study

characterized the genetic biodiversity and phylogenetic

relationships of Common Krait (Bungarus caeruleus)

from Pakistan showing inter- and intra-specific variations

among different geographical regions of the world. More

diverse sampling and a larger number of samples with

more genomic data could help to further resolve the

taxonomic status of the Bungarus species in Pakistan.

This study also provides guidance for the correct

identification of these snakes with authentication using

molecular biology tools which will be helpful in the

development of effective and region-specific antivenoms

for such venomous snakes.

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Muhammad Rizwan Ashraf is a scholar at the University of Veterinary and Animal Sciences,

Lahore, Pakistan. Muhammad has an M.Sc. in Biochemistry and Ph.D. in Molecular Biology and

Biotechnology. His research interest is reptilian (especially snake) molecular phylogeny and genetic

biodiversity. Muhammad visited University of Texas at Arlington, Texas, USA as a research scholar

under the International Research Support Initiative Program, funded by the Government of Pakistan,

and his Ph.D. work involved four of the most venomous snakes of Pakistan: Black Cobra, Common

Krait, Saw-scaled Viper, and Russell’s Viper. He is seeking further challenges to expand his skills

within a progressive and fast-growing environment that uses state-of-art technologies while enhancing

his proven abilities and dedication to team motivation.

Asif Nadeem is an Associate Professor at the University of Veterinary and Animal Sciences, Lahore,

Pakistan. Asif had the privilege of receiving research training from the University of Wisconsin—

Madison, Wisconsin, USA, and he has maintained a vigorous research program in the genetics of

animals of agricultural importance. His research has generated many publications in peer-reviewed

journals, and he has presented his research work at various national and international forums. Dr.

Nadeem’s efforts were recognized by his receipt of the Research Productivity Award from the Pakistan

Commission of Science and Technology. He is an internationally known researcher in the field of

animal genetics and genomics, and has served as an editor and reviewer for many different scientific

December 2019 | Volume 13 | Number 2 | e205

Amphib. Reptile Conserv.

Ashraf et al.

Eric Nelson Smith is a Professor at University of Texas at Arlington, Texas, USA. Eric’s current

research interest focuses on the Exploration and Speciation in the Volcanoes of the Indonesian Ring

of Fire: a Large-Scale Inventory of the Herpetofauna of the Highlands of Sumatra and Java. As a

benefit to the scientific community, this project is producing modern specimen repositories in the two

participating countries and web-based resources for identification and conservation, as well as for

genetic and biodiversity work. Many of these efforts have been directed toward the systematics and

taxonomy of venomous snakes. Eric has participated in the Threatened Amphibians of the World project

(https://portals.iucn.org/library/node/9186) and the International Union for the Conservation of Nature

(IUCN) Red List Assessment of Reptile Species, particularly with species of Asian coral snakes.

Maryam Javed is an Assistant Professor at the University of Veterinary and Animal Sciences, Lahore,

Pakistan. She has received many academic and scientific awards, including a gold medal in D.V.M.., Star

Laureate Award, Nestle Award, Tufail Muhammad Award, Best Student in Academics Award from the

Chancellor, Governor of Punjab, and the all Pakistan Quid Talent Award. Maryam’s current research

interest focuses on the identification of genes of economic importance in dairy animals.

Utpal Smart hails from Pondicherry, a sleepy coastal town (of Life of Pi fame) in southern India. Utpal’s

formal training in biology began with an M.Sc. in Ecology from Pondicherry University, India, in 2008,

followed by a Ph.D. in Quantitative Biology from the University of Texas at Arlington, USA, in 2016. His

doctoral training primarily involved using computational methods to investigate questions in molecular

ecology using a combination of macro- and micro-evolutionary approaches. As a postdoctoral research

associate at University of North Texas Center for Human Identification (UNTCHI), Utpal is helping to

create the Mitochondrial Mixture Database and Interpretation Tool (MMDIT)—a bioinformatic pipeline

for deconvoluting mitochondrial DNA mixtures and using computational phylogenetic and population

genetic methods on human microbiome data to leverage them as forensic tools.

Tahir Yaqub is a Senior Member of the University of Veterinary and Animal Sciences (UVAS), Lahore,

Pakistan, and has over 25 years of scientific experience in controlling infectious diseases of livestock,

including those caused by influenza viruses. Tahir did a postdoc at the Institute of Public Health, United

Kingdom, and his research interests include investigating the biological health risks of various agents in

public and animal health. His laboratory has postgraduate students with expertise in molecular biology

and serves as a hub for training. Currently, Tahir is investigating the prevalence and control of various

diseases of public health importance.

Abu Saeed Hashmi, Ph.D., is a well-known academician, researcher, and mentor in the field of

Biochemistry. Abu’s major area of research is in the bioconversion of agricultural/industrial waste to

value added products. Abu has supervised many postgraduate students and has contributed significantly

to this field. His work on bioconversion, Alfa toxins, bio-wastes, and production of biomass has been

published and widely cited in many prominent research journals.

Panupong Thammachoti graduated from the University of Texas at Arlington, USA, in the Quantitative

Biology Program (Ecology and Systematics). Panupong’s work used multiple approaches including

morphology, molecular phylogeny, and ecology, for solving taxonomic problems. As a lecturer at

Chulalongkorn University, Thailand, he is interested in several research topics focusing on the diversity

of amphibians and reptiles. Panupong has several scientific recognitions, such as a scholarship from the

Human Resource Development in Science Project (Science Achievement Scholarship of Thailand, SAST);

International Training Course-New Trends and Methodology in Animal Ecology and Conservation

Biology; International Society of Zoological Sciences, Beijing, China; The Professor Dr. Tab Nilanidhi

Foundation Award for outstanding academics, Faculty of Science, Chulalongkorn University; and the

best oral presentation award, JSPS CORE-TO-CORE PROGRAM at the 5th International Symposium

on Asian Vertebrate Species Diversity, Thailand.

211 December 2019 | Volume 13 | Number 2 | e205

Official journal website:

amphibian-reptile-conservation.org

Amphibian & Reptile Conservation

13(2) [General Section]: 212-216 (e206).

Daboia russelii (Reptilia: Squamata) in remote parts of Gujjar

Village Miandam, Swat, Khyber Pakhtunkhwa, Pakistan

12,*Wali Khan and 2Bashir Ahmad

'Department of Zoology, University of Malakand lower Dir, Khyber Pakhtunkhwa, PAKISTAN *Department of Zoology, Hazara University,

Mansehra, PAKISTAN

Abstract.—Snakes are widely perceived with fear by the general public in Pakistan, and they are often killed

on sight. The present study examines the range extension of Daboia russelii in remote parts of Gujjar village

Miandam Swat, Pakistan. Seven snakes were collected, including three which were attacked and injured by

the local men, and four others observed in the natural habitats in four localities: Karoo, Kalandori, Chharr, and

Dhop, from June to September in both 2016 and 2017. Morphometric analysis, details of the coloration, and

photographs of the snakes are provided. Russell’s Vipers were seen frequently in grasslands, cultivated fields,

and areas near human residences. These snakes were mostly seen after sunset. This species has also been

reported from other parts of Pakistan, but the present records represent a new locality.

Keywords. Russell’s Viper, Viperidae, morphometric analysis, range extension, snake, venomous

2 2 2

Citation: Khan W, Ahmad B. 2019. Daboia russelii (Reptilia: Squamata) in remote parts of Gujjar Village Miandam, Swat, Khyber Pakhtunkhwa,

Pakistan. Amphibian & Reptile Conservation 13(2) [General Section]: 212-216 (e206).

tion 4.0 International (CC BY 4.0): https://creativecommons.org/licenses/by/4.0/], which permits unrestricted use, distribution, and reproduction in any

medium, provided the original author and source are credited. The official and authorized publication credit sources, which will be duly enforced, are

as follows: official journal title Amphibian & Reptile Conservation; official journal website: amphibian-reptile-conservation.org.

Received: 3 August 2018; Accepted: 25 March 2019; Published: 6 December 2019

Snakes are the most reviled of vertebrates, widely

perceived by the public as dangerous and harmful. The

2,700 species of snakes known to science include 375—

400 venomous species, of which approximately 200

species are considered life-threatening to man and other

animals. Snakes of the families Elapidae, Crotalidae,

and Viperidae are venomous (Durand 2004; Vidal et al.

2009).

Russell’s Viper is named as Daboia russelii in honor

of Patrick Russell (1726-1825). Daboia russelii is an

infamous venomous snake of the Old World, found in

Bangladesh, Cambodia, China, Indonesia, Myanmar,

Nepal, Pakistan, Sri Lanka, Taiwan, and Thailand

(McDiarmid et al. 1999). In Pakistan it occurs from the

Indus Valley to the Kashmir, and east to Bengal. This

snake is frequently found in Thatta District, Sindh, and

at low elevations in Punjab, but no published report is yet

available from the northwestern parts of Pakistan.

Relatively few studies have been conducted on the

fauna of Swat, and even fewer studies are associated

with the reptilian fauna of the region (Smith 1943;

Minton 1962, 1965; Mertens 1969, 1970; Khan 1982,

1984, 1985). More than fifty species of terrestrial snakes

are known from Pakistan (Khan 1980, 1982, 1997).

Khan (2002) conducted an extensive survey of different

climatic zones in Pakistan for herpetofaunal diversity.

Correspondence. * walikhan.pk@gmail.com

Amphib. Reptile Conserv.

The present study reports the existence of Russell’s

Vipers in the hilly areas of Swat Valley, Pakistan.

Village council Miandam is located at 35°03'12"N,

72°33'39"E, about 57 km from Saidu Sharif, District Swat,

Khyber Pakhtunkhwa, at an elevation of approximately

1,918 m asl. The study area (Fig. 1) falls under moist

temperate forest, thus receiving summer monsoon and

winter snow fall. Because of its cool climate and green

hillsides, the area is frequented by tourists (Forest

Working Plan 2013).

In village council Miandam, there are two sub-villages,

namely Gujjar village and Swati village. Four sites of the

Gujjar village were surveyed from June to September in

2016 and 2017 (Fig. 2). The snake specimens (Table 1)

were either collected dead following their attack by the

local people, or recorded with visual observations using

the method of Campbell and Christman (1982).

The photographs of specimens shown here (Fig.

3) were taken using a Nikon Coolpex L330 camera.

Morphometric analysis of the snake specimens collected

dead were recorded using a digital caliper (Precision

145). The specimens observed were identified with the

help of keys provided by Khan (2002). The general

characteristics of Daboia russelii specimens from the

four localities of Gujjar village Miandam, Swat (2016-

2017) are as follows:

December 2019 | Volume 13 | Number 2 | e206

Khan and Ahmad

Fig. 1. Map of Khyber Pakhtunkhwa, red circle shows the study area in the District Swat within the province.

Table 1. Records of the seven specimens observed by month and village.

2016 2017

Dhop Chharr Karoo Kalandori Dhop Chharr Karoo Kalandori

January

February

March

April

May

June live

July dead

August dead live live dead

September live

October

November

December

Amphib. Reptile Conserv. 213 December 2019 | Volume 13 | Number 2 | e206

Fig. 2. Colle

Daboia russelii in northwestern Pakistan

a Re

ction sites

Head: Longer rather than broad, distinctly wider

than neck.

Body: Stout, flattened dorsoventrally, tapering

evenly both posteriorly and anteriorly.

Snout: Bluntly pointed, snout-vent length 1,022—

1,075 mm, tail 215-223 mm.

Rostral: About twice as high as wide.

Nostril: Large, crescent shaped in large nasal

scale.

Supraocular scale: Entire, not divided.

Supralabials: 12, separated from eye by three or

four rows of small scales.

Infralabials: 14.

Anterior chin shield: Short and wide, posterior

not well differentiated from surrounding scales.

Dorsal scales: Keeled except for lowest row, 29—

31 rows at mid-body, reduction posteriorly to 23

or 21 rows, usually an anterior reduction of two

or four rows.

Ventrals: 165-173.

Total body length: 76.2 cm, tail length 15.2 cm

(16% of total body length).

Coloration:

¢ Dorsal ground color light tan to sandy.

¢ Chest net spots with black or dark brown

Amphib. Reptile Conserv.

of Daboia russelii in Guar village Miandam, Swat, KP, Pakistan. (A) Karoo, 35°3'32"N 72°33'11"E: (B)

Kaalandori, 35°3'31"N 72°32'21"E; (C) Chhar 35°3'34"N 72°33'12"E; (D) Doop, 35°3'23"N 72°32'58"E.

borders and edge creamy, these spots fused

to a greater or lesser extent, lateral series of

similar but smaller spots below which are

scattered dark flecks with light edges.

¢ Two large dark spots at base of head.

¢ A light V-shaped mark with its apex on top

of snout.

¢ Labial sides of snout mottled with brown and

cream.

¢ Belly whitish with black semilunar spots.

¢ Chin or throat white.

¢ Many scales topped with black.

The fauna and flora of Pakistan is Oriental, Palearctic,

Ethiopian, and Central Asian in nature, with many

endemic forms (Smith 1931; Khan 1980). In Pakistan

the complex of habitats is diverse, including oceans,

swamps, rivers, lakes, flood plains, arid plains, sand and

rocky deserts; tropical thorn, tropical dry deciduous,

subtropical dry, subtropical arid, subtropical pine, dry and

moist temperate subalpine forests; grassy tundra and cold

deserts. Moreover, most of the habitats are now heavily

influenced by anthropogenic activities which negatively

affect the fauna and flora of the country (Baig 1975).

The present study describes seven specimens of

214 December 2019 | Volume 13 | Number 2 | e206

Khan and Ahmad

Russell’s Vipers from four localities in the study area,

including two specimens each from Dhoop, Chhar, and

Karoo, and one specimen from Kalandori Gujjar Village

Miandam, Swat, Pakistan. Russell’s Viper is one of the

most widespread of Asiatic venomous snakes. While

surveying the literature no published records for this

species are available in Swat Valley, Pakistan. Therefore,

the present study documents the presence of this species

in the hilly areas of Swat Valley.

Literature Cited

Baig AR. 1975. Wild life habitats of Pakistan. Biological

Science Research Desert, Punjab, Pakistan. Journal of

the Bombay Natural History Society 82: 144-148.

Campbell HW, Christman SP. 1982. Field techniques for

herpetofaunal community analysis. Wildlife Research

Amphib. Reptile Conserv.

oat

Fig. 3. Photos of the dead Daboia russelii specimens from Gujjar village Miandam, Swat, KP, Pakistan. (A, B) Dorsal views, (C,

D) Fangs, (E) Black spots on ventral side, (F) Anal orifice with tail showing a zip-like structure.

Report 13: 193-200.

Durand JF. 2004. The origin of snakes. Pp. 187

In: Geoscience Africa 2004. University of the

Witwatersrand, Johannesburg, South Africa.

Forest Working Plan. 2013-2014. Forestry Sector

Project, Village Plan, Miandam, Pakistan. pp. 7-13.

Khan MS. 1980. Affinities and zoogeography of herptiles

of Pakistan. Biologia 26: 113-171.

Khan MS. 1982. An annotated checklist and key to the

reptiles of Pakistan. Part III: Serpentes (Ophidia).

Biologia 28(2): 215-254.

Khan MS. 1984. Validity of the natricine taxon

Xenochrophis sanctijohannis Boulenger. Journal of

Herpetology 18: 198-200.

Khan MS. 1985. An Interesting Collection of Amphibians

and Reptiles from Cholistan Division. Bulletin

Number 5. Botany Department, Pakistan Forest

December 2019 | Volume 13 | Number 2 | e206

Daboia russelii in northwestern Pakistan

Institute, Peshawar, Pakistan.

Khan MS. 1997. A new toad of genus Bufo from the foot

of Siachin Glacier, Baltistan, northeastern Pakistan.

Pakistan Journal of Zoology 29: 43-48.

Khan MS. 2002. A Guide to the Snakes of Pakistan.

Edition Chimaira, Frankfurt am Main, Germany. 265

Mertens R. 1969. Die Amphibien und Reptilien West-

Pakistans. Stuttgarter Beitrdge zur Naturkunde 197:

1-96.

Mertens R. 1970. Die Amphibien und Reptilien West-

Pakistans. Stuttgarter Beitrdge zur Naturkunde 216:

1-5.

populations.

Amphib. Reptile Conserv.

216

Minton SA, Anderson S. 1965. A new dwarf gecko

(Tropiocolotes) from Balochistan. Herpetological

Bulletin 21: 59-61.

Minton SA. 1962. An annotated key to the amphibians

and reptiles of Sind and Las Bela, West Pakistan.

American Museum Novitates 81: 1-21.

Smith MA. 1931. The Fauna of British India, including

Ceylon and Burma. Reptilia and Amphibia. Volume I.

Taylor and Francis, London, United Kingdom. 85 p.

Smith MA. 1943. The Fauna of British India, Ceylon and

Burma. Reptilia and Amphibia. Volume III. Taylor

and Francis, London, United Kingdom. 583 p.

Vidal N, Rage JC, Couloux A, Hedges SB. 2009. Snakes

(serpents). The Time Tree of Life 23: 390-397.

Wali Khan currently works as Assistant Professor in the Department of Zoology, University

of Malakand, Lower Dir, Khyber Pakhtunkhwa, Pakistan. Wali is interested in understanding

the helminth parasite fauna as a factor threatening the conservation of amphibian and reptile

Bashir Ahmad is currently a research scholar in the Department of Zoology, University of Hazara,

Mansehra, Pakistan, in collaboration with the Laboratory of Parasitology, Department of Zoology,

University of Malakand, Lower Dir, Khyber Pakhtunkhwa, Pakistan. Bashir is interested in

understanding the host-parasite relationships of vertebrates as a factor threatening conservation.

December 2019 | Volume 13 | Number 2 | e206

Official journal website:

amphibian-reptile-conservation.org

Amphibian & Reptile Conservation

13(2) [General Section]: 217-226 (e207).

ptile-con*

Native anuran species as prey of invasive American Bullfrog,

Lithobates catesbeianus, in Brazil: a review with new

predation records

12,*Fabricio H. Oda, *Vinicius Guerra, “Eduardo Grou, *Lucas D. de Lima,

5SHelen C. Proenga, °Priscilla G. Gambale, **Ricardo M. Takemoto, ‘Caué P. Teixeira,

‘Karla M. Campiao, and *Jean Carlo G. Ortega

'Departamento de Quimica Biolégica, Programa de Pés-graduacgdo em Bioprospec¢do Molecular, Universidade Regional do Cariri, Campus

Pimenta, 63105-000, Crato, Ceard, BRAZIL *Departamento de Quimica Bioldgica, Laboratorio de Zoologia, Universidade Regional do Cariri,

Campus Pimenta, Crato, Ceard, BRAZIL *Departamento de Ecologia, Laboratorio de Herpetologia e Comportamento Animal, Instituto de

Ciéncias Biologicas, Universidade Federal de Goids, Campus Samambaia, Goidnia, Goids, BRAZIL *Centro de Ciéncias Bioldgicas, Nucleo de

Pesquisas em Limnologia, Ictiologia e Aquicultura, Laboratorio de Ictioparasitologia, Universidade Estadual de Maringa, Maringa, Parana,

BRAZIL *Centro de Ciéncias Biologicas, Programa de Poés-graduacdo em Biologia Comparada, Universidade Estadual de Maringd, Parana,

BRAZIL ‘Universidade Estadual de Mato Grosso do Sul, Dourados, Mato Grosso do Sul, BRAZIL ‘Departamento de Zoologia, Laboratorio

de Ecologia de Interagées Antagonistas, Universidade Federal do Parana, Centro Politécnico, Curitiba, Paranda, BRAZIL *Departamento de

Ecologia, Programa de Pos-graduacgdo em Ecologia e Evolugdo, Instituto de Ciéncias Bioldgicas, Universidade Federal de Goids, Campus

Samambaia, Goidnia, Goids, BRAZIL

Abstract.—The American Bullfrog (Lithobates catesbeianus) is widely distributed throughout the world as an

invasive species, and causes negative impacts on the fauna resulting from its voracious predatory activity.

This study documents two new predation reports and reviews the previous predation reports of the American

Bullfrog on native Brazilian anurans. Twenty-one species of native anurans were recorded as American Bullfrog

prey in Brazil. A positive correlation was found between the number of native anurans preyed on by American

Bullfrog and the respective family or number of species per genus. Most of the prey species are small or

medium-sized, and the results suggest that the generalist diet and intraguild predation may have favored the

widespread establishment of the American Bullfrog.

Keywords. Amphibia, Atlantic Forest, biological invasion, conservation, global change, intraguild predation, exotic

species

Citation: Oda FH, Guerra V, Grou E, de Lima LD, Proenga HC, Gambale PG, Takemoto RM, Teixeira CP, Campiao KM, Ortega JCG. 2019. Native

anuran species as prey of invasive American Bullfrog, Lithobates catesbeianus, in Brazil: a review with new predation records. Amphibian & Reptile

Conservation 13(2) [General Section]: 217-226 (e207).

International (CC BY 4.0): https://creativecommons.org/licenses/by/4.0/], which permits unrestricted use, distribution, and reproduction in any me-

dium, provided the original author and source are credited. The official and authorized publication credit sources, which will be duly enforced, are as

follows: official journal title Amphibian & Reptile Conservation; official journal website: amphibian-reptile-conservation.org.

Received: 8 February 2017; Accepted: 23 June 2019; Published: 20 December 2019

Introduction

Biological invasions represent a major threat to natural

ecosystems and their respective biodiversity, human

health, and food security (UCN 2012). In this context,

the American Bullfrog, Lithobates catesbeianus (Shaw,

1802), is a globally widespread introduced species (Lowe

et al. 2004). It is native in North America, occurring

from eastern Canada and the central and eastern United

States to northeastern Mexico (Quiroga et al. 2015).

The introduction of L. catesbeianus in non-native

environments has direct (e.g., predation and competition)

and indirect (e.g., parasites, disease introduction, and

biotic homogenization) impacts on biodiversity (Batista

2002; Batista et al. 2015; Kiesecker and Blaustein 1998;

Kraus 2009).

Lithobates catesbeianus 1s a voracious predator whose

diet includes a wide variety of prey (Boelter and Cechin

2007; Boelter et al. 2012; Silva et al. 2009). Juveniles

feed mainly on insects (Silva et al. 2009), whereas adults

prey upon invertebrates and small vertebrates, such as

fish, reptiles, birds, and mammals (Quiroga et al. 2015).

Correspondence. ‘fabricio_oda@hotmail.com (FHO), vinicius.guerrabatista@gmail.com (VG), eduardogrou@hotmail.com (EG), lucasdu-

artelima@hotmail.com (LDL), helencassia23@hotmail.com (HCP), priscillagambale@gmail.com (PGG), takemotorm@nupelia.uem. br (RMT),

caue.cpt@gmail.com (CPT), karla_mcamp@yahoo.com. br (KMC), ortegajean@gmail.com (JCGO)

Amphib. Reptile Conserv.

December 2019 | Volume 13 | Number 2 | e207

Impact of bullfrogs on native anurans in Brazil

American Bullfrogs are considered opportunistic feeders,

also preying on amphibians, including conspecifics and

other species (Silva et al. 2011; Toledo et al. 2007).

The American Bullfrog is now established in

nearly 40 countries around the world (Frost 2019;

Kraus 2009). In Brazil, the first specimens were

introduced in 1935 for commercial exploitation at

the municipality of Itaguai, Rio de Janeiro state

(Vizotto 1984). The introduction of the American

Bullfrog for commercial frog farming was due to its

fast reproduction and greater development in captivity

compared to native species. It occurs mainly in the

southern and southeastern Brazilian states because of

its easy adaptation to the climatic conditions (Vizotto

1984). Approximately 2,000 commercial frog farms

were active in the early 1990s in Brazil, but many

closed their activities because of low profitability

(Lima and Agostinho 1988), which led to American

Bullfrog specimens being abandoned or released into

the natural environments, and consequently several

accidental invasions have occured in Brazil (Both et

al. 2011).

Populations of Lithobates catesbeianus are now

known to be present in 155 Brazilian municipalities

(Both et al. 2011; Instituto Horus 2016), a context in

which many studies have revealed the localized impacts

of its predatory activity on native anuran fauna (Batista

et al. 2015; Boelter and Cechin 2007; Boelter et al.

2012; Leivas et al. 2012; Silva et al. 2011). In addition,

global-scale studies have demonstrated trophic niche-

width shifts in bullfrog populations from both native and

invaded areas (Bissattini and Vignoli 2017), as well as the

effects of the interactions between bullfrogs and crayfish

on native amphibians (Bissattini et al. 2018, 2019; Liu

et al. 2018). However, studies summarizing data on the

predation of native anurans by American Bullfrogs have

yet to be presented; therefore, knowledge on the impact

and the native anuran species preyed upon by such an

invasive frog may benefit our understanding of their

predator-prey relationships.

Herein, the predation of Boana raniceps and

Phyllomedusa_ distincta by males of Lithobates

catesbeianus are reported, and the available literature

on the predation of native anurans by the invasive frog

L. catesbeianus in Brazil is reviewed. An overview on

the number and identities of native species reported as

prey and the potential impact of the American Bullfrog

on native anurans are provided.

Material and Methods

Bibliographic Review

An extensive literature review was conducted to find

scientific articles, natural history notes, and theses which

contain reports on the predation of native anurans by the

Amphib. Reptile Conserv.

invasive American Bullfrog Lithobates catesbeianus in

Brazil. The sources included articles or natural history

notes published in Herpetological Review (1967-2018),

Herpetological Bulletin (2008-2018), Herpetology

Notes (2008-2018), and South American Journal of

Herpetology (2006-2018). Searches were also conducted

in Web of Science using the following query: (“Rana

catesbeiana”’ OR “Lithobates catesbeianus”) AND

(“diet” OR “feeding biology” OR “predation”, applied in

the field “topic” on 30 December 2018, without applying

any filters for year or other parameters. Considering

that predation attempts would not necessarily result in a

predation event (Toledo et al. 2007), reports of predation

attempts in the field, laboratory experiments, or captivity

were not included. Masters and doctoral papers in digital

format were obtained from the library databases of

Brazilian universities (especially Universidade Estadual

Paulista and Universidade Regional de Blumenau) by

using the search terms mentioned above in the Google

search engine.

The Web of Science query resulted in 159 studies,

three of which met the criteria and were included in the

study. Eight additional predation records were selected for

inclusion in the study by searching the selected journals

(six studies) and the library databases of Brazilian

universities (two studies). Information was extracted

from each diet analysis (1.e., the diet was described

through the analyses of stomach contents or predation

records), study location, anuran prey species, geographic

range, and body size. The geographic range follows the

list of anuran species for each Brazilian federal state

and the biomes proposed in Toledo and Batista (2012).

The body sizes of anuran species follow the size values

available in Uetanabaro et al. (2008) and Haddad et

al. (2013). The spatial distribution map of Lithobates

catesbeianus invasive populations and predation reports

were generated with 155 occurrence points for American

Bullfrog in Brazil, obtained from Both et al. (2011) and

Instituto Horus (2016).

Data Analysis

The relation between the number of native anuran

Species preyed upon by the American Bullfrog and the

number of native anurans per family or genus was tested

with a Pearson correlation analysis. The numbers of

native anuran prey species per family and genus were

compiled following the Frost (2019) database. Toledo et

al. (2007) stated that a positive correlation between the

number of predation events and taxonomic richness may

be a proxy for search representativeness, by reasoning

that taxa with more species would be more frequently

predated by chance (1.e., a sampling effect). Such a

correlation could indicate the possible mechanisms of L.

catesbeianus impacts on native biota apart from search

representativeness.

December 2019 | Volume 13 | Number 2 | e207

Oda et al.

: : j

4 a Z -— |

Fig. 1. Adult Lithobates catesbeianus swallowing an adult

Boana raniceps in an artificial permanent pond within pasture

area in southern Brazil.

Results

Field Observations and New Predation Records

An adult Lithobates catesbeianus swallowing an adult

Boana raniceps (Fig. 1) was recorded on 11 October

2014 at 2100 h, in an artificial permanent pond inside a

pasture area (23°20’38”S, 51°52’07”W), in the northern

region of Parana state, southern Brazil. Although the

specimens escaped, voucher specimens of the native

anuran species and L. catesbeianus had been previously

collected by Affonso et al. (2014) and stored at the

Amphibian Collection from the Zoology and Botany

Department, Bioscience Institute, Universidade Estadual

Paulista, Rio Claro, Sao Paulo, Brazil.

A second predation event recorded a male adult

specimen of Bullfrog swallowing a treefrog (Fig. 2A).

The specimen was collected during an L. catesbeianus

survey on 22 January 2019, at 2200 h, in an artificial

permanent pond in a rural property at Iporanga, southern

Sao Paulo state, southeastern Brazil (24°35’01.2”S,

48°36’00.4”W). The L. catesbeianus specimen was taken

to the laboratory where the anuran prey was removed and

identified as an adult Phyllomedusa distincta (Fig. 2B).

removed from the oral cavity of L. catesbeianus.

Amphib. Reptile Conserv.

Exploratory Analysis

Overall, 11 publications reported predation events,

corresponding to 41 records of native anurans as prey

of L. catesbeianus (Table 1). Nine of the publications

discussed the diet in a broader sense, and two were

natural history notes reporting predation events. Most

of the records occurred in Minas Gerais state (39%),

followed by Rio Grande do Sul (~32%), Parana (~12%),

Sao Paulo (~12%), and Santa Catarina (~5%), at sites

inside the Atlantic Forest, in addition to another site in

a transition zone between Cerrado and Atlantic Forest

(Fig. 3, Table 1).

This survey accounted for 21 anuran species as prey

of L. catesbeianus, all widely distributed and possibly

coexisting with American Bullfrogs in their breeding

sites. The anuran family Hylidae had the highest number

of species (11 species), followed by Leptodactylidae (four

species), Bufonidae and Microhylidae with two species

each, and Odontophrynidae and Phyllomedusidae with

one species each. Lithobates catesbeianus often preyed

on medium-sized species, but small-sized species were

also preyed upon (Table 1).

A positive correlation was found between the number

of native anuran species preyed on by American Bullfrog

and genus richness (r = 0.71, P = 0.01), whereas at the

family level no relationship was found (r = 0.53, P =

0.22). Thus, considering the studies analyzed, genera

with higher numbers of species presented more potential

prey for American Bullfrogs in Brazil.

Discussion

Most of the predation records found in this review came

from a few studies which assessed the overall dietary

composition of L. catesbeianus, and revealed that the

diet of these invasive frog populations is represented

by a wide variety of native anuran species (Boelter and

Cechin 2007; Silva et al. 2009, 2010, 2011). Only two

predation records of L. catesbeianus and native anurans

in the field were found, probably due to some difficulty

cal

Fig. 2. (A) Predation of an adult Phyllomedusa distincta by Lithobates catesbeianus, (B) Adult P. distincta partially digested,

December 2019 | Volume 13 | Number 2 | e207

Impact of bullfrogs on native anurans in Brazil

0°

10° S$

20°S

Biomes

[ Amazon

[ Caatinga

[| Cerrado

Atlantic Forest

| Pampa

[| Pantanal

30° S

70° W 60° W

50° W 40° W

Fig. 3. Spatial distribution of Lithobates catesbeianus invasive populations and predation reports of native anurans in Brazil. White

circles: American Bullfrog populations in Brazil (Both et al. 2011; Instituto Horus 2016); yellow stars: predation reports of adult

Boana raniceps and adult Phyllomedusa distincta in southern and southeastern Brazil; light green circles: locations of 41 published

predation records.

in recording and quantifying these events in the field

(Pombal Jr. 2007).

The predicted potential occurrence of L. catesbeianus

in Brazil represents its current distribution in the southern

and southeastern regions in the Atlantic Forest, with

potential areas for colonization remaining in the central

and northeastern regions (Giovanelli et al. 2008; Both et

al. 2011). The results showed that all predation records

occurred at sites in southern and southeastern Brazil,

regions with higher numbers of research centers, thus

contributing a disproportionately greater number of field

studies.

Native anurans recorded as prey of American

Bullfrogs share the same breeding sites. Silva et al.

(2011) had found a spatial overlap in microhabitat use

between native species and American Bullfrogs during

the reproductive season. American Bullfrogs may also

overlap with native amphibians in diet composition

Amphib. Reptile Conserv.

(Bissattini et al. 2019). This may lead to a potential

competition, and may have a direct influence on

community composition patterns since the intrinsic

ecological properties of organisms determine the niche

overlap between species in the communities (Vignoli

and Luiselli 2012; Vignoli et al. 2017). Additionally, the

predation on other anuran species by L. catesbeianus can

represent an example of intraguild predation (Polis et al.

1989), a process that may facilitate the establishment of

the American Bullfrog (Bissattini et al. 2018), as found

in other disparate introduced taxa, such as ladybird

beetles (Snyder et al. 2004) and fish (Pereira et al. 2015).

Intraguild predation can benefit the establishment of L.

catesbeianus by reducing the competitive pressure by

direct predation of the other anuran species.

The number of prey species had a positive correlation

with the number of species per genus, in which the

family Hylidae had the highest number of species as

December 2019 | Volume 13 | Number 2 | e207

Oda et al.

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December 2019 | Volume 13 | Number 2 | e207

222

Amphib. Reptile Conserv.

Oda et al.

prey of L. catesbeianus. Boelter et al. (2012) found

that 60% of the prey records corresponded to Hylidae

Species, suggesting that this group suffers higher

predation pressure. At least two non-mutually exclusive

hypotheses may explain these patterns by relating family

(i.e., richness, abundance) and species traits (1.e., body

size) to L. catesbeianus predation rates, involving

species traits that are often phylogenetically correlated

(Martins and Hansen 1997). Firstly, it is possible that

Hylidae species are often preyed upon due to their higher

species richness in comparison to other families, which

is a plausible hypothesis if we assume that predation

rates can be proportional to prey abundance or richness

(i.e., higher predation rates in higher resource availability

conditions; Jacobsen et al. 2014; Madahi et al. 2015).

Secondly, Hylidae species may have a higher predation

rate because of their smaller size relative to species from

other families (e.g., Bufonidae). Predators that feed

on whole animals, such as the American Bullfrog, are

limited by the prey’s body size. Experimental evidence

indicates that larger specimens of L. catesbeianus feed

preferentially on smaller, rather than on large-sized,

native anurans (Wang et al. 2007), suggesting a size-

based selection of prey species. Partially related to this

hypothesis, our results suggest a higher amount of small

and medium-sized species as American Bullfrog prey.

Therefore, the preference for prey of a certain body size

may be proportionally related to the body size of the

predators, as observed in previous studies (Quiroga et al.

2015; Silva et al. 2011, 2009; Wang et al. 2007).

This survey found that all anuran species preyed upon

by L. catesbeianus have large geographic distributions,

occurring in various Brazilian states and biomes (Frost

2019; Toledo and Batista 2012). Both et al. (2014)

found that American Bullfrog abundance had a positive

relationship with communities that consisted of generalist

species (e.g., Physalaemus cuvieri, Dendropsophus

minutus), that were anthropogenically adapted and

broadly distributed in South America. Native anurans

with large geographic ranges (e.g., Rhinella diptycha,

Dendropsophus minutus, Boana faber, B. raniceps,

Scinax fuscovarius, and Physalaemus cuvieri) have also

been found in sympatry with Lithobates catesbeianus

elsewhere (Affonso et al. 2014).

Conclusions

This study indicated L. catesbeianus preys on at least

21 native anuran species in Brazil. Predation is one of

the major negative effects of Invasive species on native

communities. The quality of being a generalist feeder,

preying on many anuran species, has benefited the

successful colonization, establishment, and permanence

of the American Bullfrog in Brazil (and even worldwide;

e.g., Li et al. 2011; Monello et al. 2006; Quiroga et al.

2015). Native species of the family Hylidae may be more

susceptible to American Bullfrog predation because of

Amphib. Reptile Conserv.

their higher abundance and richness, and/or due to a

higher representation of small- to medium-sized species

relative to other anuran families. Knowledge on the

species most vulnerable to predation by the American

Bullfrog can enable better prediction of the negative

impacts of such an invasive species on native anuran

communities.

Acknowledgements.—The authors would like to thank

Centro Universitario de Maringa for having granted us

access to the Fazenda Escola UniCesumar, and Bruno

Barreto for assisting with Fig. 3. Fabricio H. Oda received

a postdoctoral fellowship from Funda¢ao Cearense de

Apoio ao Desenvolvimento Cientifico e Tecnoldgico/

Coordenacéo de Aperfeicoamento Pessoal de Nivel

Superior — CAPES (Grant number 88887.162751/2018-

00). Vinicius Guerra received fellowships from CAPES,

and Helen C. Proenca and Jean Carlo G. Ortega received

fellowships from Conselho Nacional de Desenvolvimento

Cientifico e Tecnologico — CNPq. Ricardo M. Takemoto

received a CNPq grant of research productivity

fellowship as well. Finally, the authors would like to

thank the Instituto Chico Mendes de Conservacao da

Biodiversidade/Sistema de Autorizacéo e Informacao

em Biodiversidade for providing us with the collecting

permit (process #23866-1).

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Fabricio Hiroiuki Oda is a postdoctoral research fellow in the Programa de Pos-graduacéo em

Bioprospec¢ao Molecular (URCA — Universidade Regional do Cariri, Brazil) and research collaborator

in the Laboratorio de Ictioparasitologia do Nucleo de Pesquisas em Limnologia, Ictiologia e Aquicultura

(UEM — Universidade Estadual de Maringa, Brazil). His primary areas of interest are the natural history,

ecology, and parasitology of fish, amphibians, and reptiles.

Vinicius Guerra is a postdoctoral researcher in the Laboratorio de Herpetologia e Comportamento Animal

(UFG — Universidade Federal de Goias, Brazil). His primary areas of interest are the study of community

ecology, animal behavior, natural history, and bioacoustics, with a particular focus on amphibians.

Eduardo Grou is a volunteer researcher in the Laboratorio de Ictioparasitologia do Nucleo de Pesquisas em

Limnologia, Ictiologia e Aquicultura (UEM — Universidade Estadual de Maringa, Brazil). His primary areas

of interest are the ecology and parasitology of chelonians.

December 2019 | Volume 13 | Number 2 | e207

Impact of bullfrogs on native anurans in Brazil

Lucas Duarte de Lima is a Master’s student fellow in the Programa de Pos-gradua¢ao em Biologia Comparada

(UEM — Universidade Estadual de Maringa, Brazil). His primary area of interest is amphibian diversity.

Helen Cassia Proenca is a Doctoral student fellow in the Programa de Pos-graduacao em Biologia Comparada

(UEM — Universidade Estadual de Maringa, Brazil). Her primary area of interest is snake diversity.

Priscilla Guedes Gambale is a researcher in the Departamento de Biologia (UEMS —Universidade Estadual

de Mato Grosso do Sul, Brazil). Her primary area of interest is the ecology of amphibians, with a particular

focus on bioacoustics.

Ricardo Massato Takemoto is a researcher in the Laboratorio de Ictioparasitologia do Nucleo de Pesquisas

em Limnologia, Ictiologia e Aquicultura (UEM — Universidade Estadual de Maringa, Brazil) and advisor in

the Programa de Pés-gradua¢4o em Ecologia de Ambientes Aquaticos Continentais and the Programa de Pos-

gradua¢ao em Biologia Comparada (UEM — Universidade Estadual de Maringa, Brazil). His primary area of

interest is the study of parasites of aquatic organisms, including fish and amphibians.

Caué Pinheiro Teixeira is Master’s researcher in the Laboratorio de Ecologia de Interagées Antagonistas

(UFPR — Universidade Federal do Parana, Brazil). His primary areas of interest are ecology, herpetology,

parasitology, and biological invasion.

Karla Magalhaes Campiao is a researcher and coordinator of the Laboratorio de Ecologia de Interagdes

Antagonistas (UFPR — Universidade Federal do Parana, Brazil), and she supervises graduate students in

Ecology and Zoology. Her primary areas of interest are amphibian parasites and disease ecology.

Jean Carlo Goncalves Ortega is a postdoctoral researcher in the Programa de Pos-graduacéo em Ecologia

e Evolugaéo (UFG — Universidade Federal de Goias, Brazil). His primary areas of interest are community

ecology and biological invasions.

Amphib. Reptile Conserv. 226 December 2019 | Volume 13 | Number 2 | e207

Official journal website:

amphibian-reptile-conservation.org

Amphibian & Reptile Conservation

13(2) [General Section]: 227-238 (e208).

Biology of snakes of the genus Tretanorhinus:

an integrative review

12.*Mlarco D. Barquero and ‘*Viviana Arguedas

'Asociacion para la Conservacion y el Estudio de la Biodiversidad (ACEBIO), San José, COSTA RICA *Sede del Caribe, Universidad de Costa

Rica, Montes de Oca, San José, 2060, COSTA RICA +Sede de Occidente, Universidad de Costa Rica, Montes de Oca, San José, 2060, COSTA RICA

Abstract.—Many aspects of the biology of various snake species remain unknown, and the extent of this lack

of information is not always clear. As new research usually depends upon previous findings, the gaps in our

knowledge and the accuracy of published information are of major importance. Therefore, an analysis of all

available information on snakes of the genus Tretanorhinus, from both the literature and museum specimens,

is presented here to illuminate existing knowledge gaps. The database compiled from 87 documents referring

to snakes of this genus and 755 specimens held in scientific collections revealed major gaps and contradictory

information for all four species of this genus. Data on morphology, ecology, and natural history are completely

absent for 7. mocquardi and T. taeniatus, whereas confusing distribution reports exist for T. nigroluteus. The

potential consequences of these problems were determined, and some suggestions for correcting them are

addressed. Specifically, we consider that focused efforts on the validation of current species and subspecies,

field and lab studies of ecology and behavior, and estimations of population dynamics, are necessary.

Keywords. Colubridae, Dipsadidae, ecology, literature review, morphology, museum specimens, natural history, Rep-

tilia, Serpentes, taxonomy

Citation: Barquero MD, Arguedas V. 2019. Biology of snakes of the genus Tretanorhinus: an integrative review. Amphibian & Reptile Conservation

13(2) [General Section]: 227-238 (e208).

[Attribution 4.0 International (CC BY 4.0): https://creativecommons.org/licenses/by/4.0/], which permits unrestricted use, distribution, and reproduction

in any medium, provided the original author and source are credited. The official and authorized publication credit sources, which will be duly enforced,

are as follows: official journal title Amphibian & Reptile Conservation; official journal website: amphibian-reptile-conservation.org.

Received: 24 June 2018; Accepted: 6 December 2018; Published: 10 December 2019

Introduction

Fieldwork with snakes usually poses several challenges

and more attention has focused on snake species from

temperate areas than those from the tropics (Avila et al.

2006), despite the latter having higher diversity (Greene

1997). This bias has resulted in a lack of key information

on essential aspects of the ecology, natural history, and

behavior of many species. The collection of such data

is a time-consuming and difficult task, as many snake

species have cryptic habits and occur in low densities

(Greene 1997). However, the lack of basic information

for many snake species is a major concern, because this

information is crucial for making general interspecific

comparisons, establishing proper phylogenetic

relationships, determining population trends and

dynamics, and recognizing differences in behavior.

Therefore, reviewing the current state of knowledge

for a given species is imperative to identify the areas

that require more attention and to guide research and

conservation efforts in the proper direction.

Snakes of the genus Tretanorhinus are nocturnal,

aquatic species that have intrigued biologists for more

than a century (Cope 1861; Dumeéril et al. 1854). Their

habits and secretive behavior make them difficult to

study, so their biology remains largely unknown (Savage

2002; Schwartz and Henderson 1991). Currently, four

species are recognized in the genus: 7’ mocquardi

(Bocourt 1891), 7. nigroluteus (Cope 1861), 7: taeniatus

(Boulenger 1903), and 7: variabilis (Duméril et al.

1854). Although a handful of studies have increased

our knowledge about these species (e.g., Barquero et al.

2005; Dunn 1939; Henderson and Hoevers 1979; Villa

1970), the genus remains poorly studied.

Here we summarize and integrate all available

published information, highlight the gaps in our

knowledge of the biology of Tretanorhinus species,

and make suggestions to direct future research in

order to fill in these gaps. To achieve this, searches

were conducted to find all published material referring

directly (1.e., studies focused specifically on one or more

Tretanorhinus species) or indirectly (1.e., studies focused

on many taxa that mentioned one or more 7retanorhinus

Species as part of the topic) to snakes of this genus and

a database of key information was compiled (Appendix

1). The information mentioned in each study was

used to determine missing data for each species. Each

publication was classified using the following categories:

Correspondence. !** marco.barquero_a@ucr.ac.cr, | viviarguedas@gmail.com

Amphib. Reptile Conserv.

December 2019 | Volume 13 | Number 2 | e208

Biology of Tretanorhinus species

—o mocquardi

=H nigroluteus

—®- taeniatus

—i-variabilis

Number of individuals

T T T T T T T T T ® T

Jan Feb Mar Apr May Jun Jul Aug Sep Oct Nov Dec

Fig. 1. Number of specimens of the four species of Tretanorhinus

according to (A) decade and (B) month of collection.

natural history, morphology, taxonomy, systematics,

ecology, distribution, biogeography, and reproduction.

In addition, data were obtained for specimens held

in scientific collections (Appendix 2) from the

HerpNet2 data portal (http://www.herpnet.org), Global

Biodiversity Information Facility (http://www. gbif.org),

Data Research Warehouse Information Network (http://

darwin.naturalsciences.be), and by directly contacting

collection curators. When available, the following data

were extracted: taxonomic classification, type status,

sex and age classes, country and locality of the point

of capture, latitude and longitude of the collection site,

date of collection, and remarks about the individual

collected.

Literature Review and Scientific Collections

The search for publications referring to 7retanorhinus

produced a total of 87 documents, although only 16

focused directly on species of the genus (Appendix

1). Most of the documents focused on general aspects

of natural history (31%), taxonomy (21.8%), and

biogeography (20.7%), and the vast majority referred

to T. nigroluteus or T: variabilis, with only a handful of

studies mentioning 7. mocquardi (n = 12) or T: taeniatus

(n= 8).

A database with information for a total of 755

specimens of Tretanorhinus held in 31 scientific

collections was compiled (Appendix 2). Most records

corresponded to 7: nigroluteus (n = 350) or T: variabilis

(n= 357), and only a few were available for 7. mocquardi

(n= 25) or 7. taeniatus (n = 9). In addition, eight records

Amphib. Reptile Conserv.

were identified only to the genus level, and six were

mistakenly identified as Tretanorhinus agassizi. Most of

the collections occurred between the 1940s and 1970s

(Fig. 1A) and seasonally between June and August (Fig.

1B), although many records did not include either the

year or month of collection.

Distribution and Habitat

Tretanorhinus 1s a genus exclusive to Middle and South

America, and the West Indies. 7retanorhinus variabilis

is the only one of the species with an unambiguous

distribution, inhabiting Cuba, Isla de la Juventud (Isle of

Youth), and the Cayman Islands (Fig. 2). Tretanorhinus

nigroluteus has been frequently recorded along the

Atlantic coast from southern Mexico to Colombia,

including some islands such as the Bay Islands of

Honduras and Great Corn Island of Nicaragua (Fig.

2). However, records of this species in Colombia are

controversial. Alarcon-Pardo (1978) reported finding 7.

nigroluteus on the Atlantic coast of Colombia, but we

were unable to locate any specimens of this species in

the scientific collections consulted which matched the

locality mentioned by that author. We found only one

specimen of 7! nigroluteus from the Pacific coast of

Colombia as a disjunct point of the species range (Table

1). The distributions of 7. mocquardi and T. taeniatus

are the least clear among the species of the genus.

Tretanorhinus mocquardi ranges from the Canal Zone

in Panama through the Pacific coast of Colombia and

Ecuador, although it has been reported from only two

locations in Colombia and one in Ecuador (Fig. 2, Table

1). Tretanorhinus taeniatus could be endemic to Ecuador

(Table 1), despite previous reports of this species in

Colombia (Castafio-M. et al. 2004; Daniel 1949). Based

on these distributions, 7? mocquardi and T. taeniatus

could be sympatric in Esmeraldas province, northwestern

Ecuador, whereas 7? mocquardi and T: nigroluteus seem

to be sympatric in Choco department on the Pacific coast

of Colombia (Fig. 2).

Most information on the preferred habitat of the

genus comes from studies conducted on 7! nigroluteus

and T. variabilis. Tretanorhinus 1s a fully aquatic genus,

inhabiting all kinds of fresh and brackish water bodies

such as rivers, streams, lagoons, estuaries, mangroves

(Cisneros-Heredia 2005; Neill 1958; Villa 1970), and

even cow wells (Seidel and Franz 1994). The species

require a muddy or rocky bottom with aquatic vegetation

where they can hide and rest (Neill 1965; Villa 1970).

Although there are reports of individuals found out of

the water (e.g., crossing roads), these events occur after

flooding that forces the snakes to search for another water

body (Barquero et al. 2005; Villa 1970). The distributions

of 7. nigroluteus and T. variabilis also confirm the ability

of these snakes to disperse and survive in salt water

(Barbour and Amaral 1924: Neill 1958), since they have

colonized several islands. In that regard, we observed

December 2019 | Volume 13 | Number 2 | e208

Barquero and Arguedas

Species

nigroluteus

@ faeniatus

variabiis

QO 200 400 800 1,200 1,600

a es KT

Fig. 2. Map showing the current distribution of the four species of Zretanorhinus based on specimens from scientific collections.

Table 1. Number of specimens of Tretanorhinus species collected from different countries and held in several scientific collections.

Specimens with a doubtful or missing location were excluded (n = 38).

Country T. mocquardi T. nigroluteus T. taeniatus T. variabilis Total

West Indies

Cayman Islands - - - 58 58

Cuba - - - 280 280

Middle America

Belize - 52 - - 52

Costa Rica - 11 - - 11

Guatemala - 11 - - 11

Honduras - 157 - - 157

Mexico - 30 - - 30

Nicaragua - 65 - - 65

Panama 14 19 - - 33

South America

Colombia 4 1 - - 5

Ecuador 6 - 9 - 15

Total 24 346 9 338 717

Amphib. Reptile Conserv. 229 December 2019 | Volume 13 | Number 2 | e208

Biology of Tretanorhinus species

Table 2. Morphological information available in the literature for each species of Tretanorhinus.

Trait T. mocquardi T. nigroluteus T. taeniatus T. variabilis

SVL (mm)

Adults - 242-656 440 500-800

Juveniles - 140-168 - 143-145

Tail length (mm)

Adults - 142-206 130 145-160

Juveniles - 51-77 ~ -

No. of loreals 1 1-2 1 1-3

No. of prefrontals 1-2 2 3 )

No. of preoculars 2 2-3 2 1-3

No. of postoculars - 2 2 2

No. of temporals - 1+2 BE? 243 1+2

No. of upper labials - 7-9 8 8-9

No. of lower labials - 9-12 4—5 9-11

No. of ventrals 166-177 127-151 168-175 152-168

No. of caudals 69-85 56-81 74-81 48-81

Posterior chin-shields In contact In contact Separated Separated

No. of dorsal rows 19 19, 21 21 1D: 2)

Ventral color Fuliginous yellow rane. eee yellow, - ae dats

Ventral pattern -

Dorsal color Ss

3 longitudinal

Dorsal pattern ;

stripes

an individual of 7) nigroluteus resting on a beach on

the Caribbean coast of Costa Rica after a heavy rain,

suggesting that the snake had been washed out to sea,

surviving until it was returned to land by the tide.

Morphology

Morphological information is limited and incomplete

for TZ] mocquardi and T: taeniatus, despite both species

being described more than 110 yrs ago by Bocourt

(1891) and Boulenger (1903), respectively. Table 2

provides a summary of some morphological features

that were extracted from the literature review. Overall,

Tretanorhinus are relatively small snakes; 7. variabilis is

the largest species with an SVL of up to 800 mm. All four

species in the genus exhibit a grayish dorsal coloration

with stripes, spots, or bands, and a yellow, orange, or

gray ventral coloration. Juveniles have been found in

the wild only infrequently, and the sex of most collected

individuals was not identified (Table 3). Juveniles of

each species resemble the adults in pattern and coloration

(Barquero et al. 2005; Petzold 1967).

Amphib. Reptile Conserv.

With or without dots or spots

Olive, grayish brown, light

brown, black

With or without dark spots

3 dark stripes Light dots or spots

Grayish olive, dark

Grayish olive Beaten

3 longitudinal

: Blackish cross bands

stripes

Previous attempts to produce a key to the species

of the genus have never included all four species.

Therefore, the following key is presented to unify the

previous efforts (Bocourt 1891; Boulenger 1893; Dunn

1939; Peters and Orejas-Miranda 1970; Kohler 2008)

and incorporate more recent data.

Key to the species of 7retanorhinus:

la. Dorsum without stripes, more than one loreal can be

present. «7.2. 8.4.. BONE Shley ie

1b. Dorsum: with three longitudinal stipes. only one Joreal

2a. Posterior chin-shields in contact, ventrals fewer than

152. Focad ae nigroluteus

2b. Posionet chinshields ‘Separated ‘wenthals 152 or

more. He. ots eke, VALLADALS

3a. Less ‘than three ptefrontals. posterior chin- shields in

contact.. ay : ..mocquardi

3b. Three preftontals, posterior chin-shields separated

..taeniatus

December 2019 | Volume 13 | Number 2 | e208

Barquero and Arguedas

Taxonomy and Systematics

The evolutionary relationships of the genus are enigmatic

and controversial, so that 7retanorhinus has been placed

within different former subfamilies of Colubridae (e.g.,

Natricinae, Xenodontinae, and Dipsadinae) by various

authors (Crother 1999; Dowling et al. 1996; Pinou and

Dowling 1994; Villa 1969). The general consensus 1s that

the genus should be placed within Dipsadidae (formerly

Dipsadinae, Grazziotin et al. 2012; Pinou and Dowling

1994) and it is commonly referred to as a xenodontine

(Cadle 1985; Minton 1976; Vidal et al. 2000). Moreover,

the phylogenetic relationships with other genera are still

ambiguous. Crother (1999) placed Tretanorhinus as the

sister taxon of Sibon or the Sibon-Manolepis clade. In

two studies that included more taxa, Grazziotin et al.

(2012) noted that Tretanorhinus could be the sister taxon

of Hypsiglena, and both genera were placed in a clade

containing 7rimetopon, Geophis, and Atractus, whereas

Pyron et al. (2013) placed Tretanorhinus as the sister

taxon of the Leptodeira-Imantodes clade. However, in all

these studies the relationships of Tretanorhinus with other

taxa were poorly supported and the genus is considered as

Dipsadidae incertae sedis (Grazziotin et al. 2012).

The affinities of each species within Tretanorhinus

are also enigmatic, and this is caused by the lack of

information on the basic biological aspects of some

species. Detailed morphological information is only

available for 7. nigroluteus and T. variabilis (Table 2). No

author has given a complete morphological description

of 7’ mocquardi, such that even basic phenotypic traits

(e.g., most head scutellation and dorsal coloration)

are missing (Table 2). In the case of 7 taeniatus,

morphological information has only been reported for

females, as males have never been deposited or identified

as such in scientific collections (Table 3). Despite these

deficiencies, morphological information compiled from

the literature review here suggests that 7. taeniatus and 7:

mocquardi are more closely related to each other than to

7. nigroluteus and T: variabilis.

Problems have also arisen with the subspecies

described for both TZ nigroluteus (dichromaticus,

lateralis, mertensi, nigroluteus, and obscurus) and T.

variabilis (binghami, insulaepinorum, lewisi, variabilis,

and wag/eri). The morphological variables used to define

the subspecies have been chosen arbitrarily (Wilson

and Hahn 1973). For example, differences in coloration

have been used by some authors to differentiate among

subspecies with only superficial descriptions (Schwartz

and Ogren 1956; Smith 1965; Villa 1969). In addition,

no genetic analyses have yet been reported to confirm the

validity of these subspecies.

Ecology and Natural History

Information on the ecology and natural history of 7.

mocquardi and T. taeniatus is virtually non-existent

Amphib. Reptile Conserv.

Table 3. Number of specimens of Tretanorhinus held in

scientific collections that have been identified as female, male,

or juvenile.

Species Female Male Juvenile Total

T. mocquardi 2 1 - S)

T. nigroluteus 14 16 2 32

T. taeniatus 2 - - 2

T: variabilis 19 17 4 40

Total 37 34 6 77

and they remain poorly studied for 7. nigroluteus and T.-

variabilis. Snakes of Tretanorhinus are nocturnal and seem

to hide during the day in water bodies amongst roots and

rock crevices (Barbour and Ramsden 1919; Stuart 1937;

Villa 1970). Some unusual features have been identified

for the genus. For example, individuals of Tretanorhinus

feed upon fishes, tadpoles, and frogs by either actively

chasing prey or remaining motionless with the tail and

body attached to a supporting surface (e.g., branch or rock)

and striking at passing prey (Barquero et al. 2005; Neill

1965; Petzold 1967; Wilson and Hahn 1973). In addition,

these snakes demonstrate shy behavior, such as fleeing to

the bottom of water bodies when disturbed (Stuart 1937)

and rolling up the body like a ball when caught (Petzold

1967; Seidel and Franz 1994). Known natural predators

of Zretanorhinus include, but are probably not limited

to, turtles (e.g., Kinosternon [Villa 1973]) and wading

birds (e.g., Tigrissoma and Cochlearius [Villa 1970]).

A specimen from Costa Rica was collected from a crab,

thus confirming the assumption of Villa (1970) that some

species of crabs can be predators of Tretanorhinus.

Information on reproduction is scarce for all four

species of Tretanorhinus, although observations in

captivity demonstrate that 7’ nigroluteus and T: variabilis

are oviparous, laying 6—9 adherent eggs out of water

(Barquero et al. 2005; Petzold 1967). Tretanorhinus

variabilis lays larger eggs (35[L] x 16.75[W] mm on

average) that hatch earlier (35 d) than 7) nigroluteus

(21.5[L] x 10[W] mm, 42 d). Villa (1970) found gravid

females of 7. nigroluteus during both the dry and wet

seasons, suggesting that reproduction could occur

year-round in this species. Most gravid females of 7°

variabilis have been found during the wet season in July

and August (Petzold 1967; Seidel and Franz 1994). Both

T! nigroluteus and T. variabilis are sexually dimorphic,

with females being larger than males and only males

possessing tubercles on scales of the head (Henderson

and Hoevers 1979; Petzold 1967).

The capacity to survive in salt water likely contributed

to the colonization of Caribbean islands and northeastern

South America from a Central American ancestor

(Cisneros-Heredia 2005; Hedges 1996) and allowed 7.

nigroluteus and T! variabilis to become fairly abundant in

some parts of their ranges (Henderson and Hoevers 1977;

Schwartz and Henderson 1991). Henderson and Hoevers

(1977) reported that 7. nigroluteus was more frequently

December 2019 | Volume 13 | Number 2 | e208

Biology of Tretanorhinus species

found during the dry season (December to May) than

during the wet season (June to November) of Belize,

a difference explained by the overflowing of the river

system studied. However, historical collections of this

species show that more individuals have been collected

during the wet season, a pattern shared with T. variabilis

(Fig. 1), and in accordance with reports by Wilson and

Hahn (1973) for Roatan Island, Honduras. 7retanorhinus

mocquardi and T: taeniatus are less abundant than the

other two species and no pattern of variation in abundance

can be elucidated from available data.

General Considerations and Future Research

This study has summarized information about the

species of the genus Tretanorhinus published from

1854 (Dumeril et al. 1854) to the present (Estrella-

Morales and Piedra-Castro 2018). Information was also

incorporated on all collected specimens of this genus that

could be identified as preserved in scientific collections

throughout the world. This integrative approach allowed

the identification of gaps in our knowledge about these

snakes. For example, it is surprising that (1) a complete

morphological description is not available in the literature

for 7’ mocquardi, (2) only a few specimens have been

collected for 77] mocquardi and T. taeniatus, and (3) most

natural history and ecological information simply has

never been reported for any of the species. In addition

to the lack of key data, much available information is

contradictory, such as the reported occurrence of 7

nigroluteus and T: taeniatus in Colombia, which can

cause several problems. Therefore, one can ask how a

reliable identification of individuals in the field can

be made when such basic data are missing. This is

particularly problematic for sympatric species, such as

7. nigroluteus and T: mocquardi in Panama, and for 7:

mocquardi and T: taeniatus in Ecuador.

In order to fill in the gaps in our knowledge of

Tretanorhinus, future research should focus on at least

three areas. First, validation of the currently accepted

species and subspecies 1s absolutely urgent. Barbour

and Amaral (1924) have questioned the validity of 7.

mocquardi (although see Dunn 1939), while Wilson and

Hahn (1973) refused to recognize 7: n. dichromaticus.

Therefore, a comparative study of all species and

subspecies that includes both morphological and genetic

data and produces a phylogeny of the genus is necessary.

Previous attempts have failed to include all species or

have used only morphological or genetic data. Second,

both field and lab studies are needed to increase our

knowledge about these secretive and, in some areas,

elusive species. Tretanorhinus mocquardi and T.

taeniatus require extensive work on morphological

variation, ecological habits, distribution patterns, and

natural history traits. Breeding behavior is completely

unknown for these two species and males of 7) taeniatus

have yet to be measured and described. Although

Amphib. Reptile Conserv.

there is significantly more information available for 7:

nigroluteus and 7: variabilis, nothing is known about

their courtship, sexual selection, development, and

many other aspects of basic biology. Third, demographic

variation and population dynamics need to be quantified

to understand the movement of individuals among

populations, sex ratios, and population sizes. These types

of data are essential for determining the conservation

status of species. Some efforts have already been made

to identify areas of high and low abundance across the

ranges of 7: nigroluteus and T: variabilis. However, long-

term studies which monitor changes in populations are

yet to be done.

The problems mentioned above are not restricted only

to Tretanorhinus species, as they also apply to many other

snakes (Dorcas and Willson 2009), and it is alarming

that we rely so heavily on unconfirmed or erroneous

information. Snakes in particular require special attention

due to the intrinsic difficulties in generating accurate

information. These difficulties arise from certain aspects

of their biology, including low densities, great mobility,

and cryptic habits. Integrative studies, such as this one,

are important for identifying the gaps in our knowledge

of different taxa and guiding future efforts in the right

direction.

Acknowledgements.—We are grateful to all the curators

at institutions that kindly provided us with the information

on specimens of 7retanorhinus. We are also thankful to

Pablo Allen, Guido Saborio, and Rowan McGinley for

their helpful comments on the manuscript.

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Biology of Tretanorhinus species

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Marco D. Barquero is a Costa Rican biologist with a Doctorate degree from Macquarie

University, Sydney, Australia. Marco has dedicated the last 18 years to the study of amphibians

and reptiles in different parts of the world, but especially in Costa Rica.

Viviana Arguedas is a Costa Rican biologist with a Master’s degree from the University of Costa

Rica. Viviana has studied different taxa over the last 14 years, with an emphasis on herpetofauna.

Appendix 1. List of studies referring to 7retanorhinus species.

1. Alarcon-Pardo H. 1978. Primer registro de T7retanorhinus nigroluteus nigroluteus Cope (Reptilia: Serpentes:

Colubridae) para Colombia. Lozania 27: 1-4.

2. Auth DL. 1994. Checklist and bibliography of the amphibians and reptiles of Panama. Smithsonian Herpetological

Information Service 98: 1-59.

3. Barbour T. 1914. A contribution to the zoogeography of the West Indies, with special reference to amphibians and

reptiles. Memoirs of the Museum of Comparative Zoélogy at Harvard College 44: 207-359.

4. Barbour T. 1916. The reptiles and amphibians of the Isle of Pines. Annals of Carnegie Museum 10: 207-308.

5. Barbour T; Amaral AD. 1924. Notes on some Central American snakes. Occasional Papers of the Boston Society of

Natural History 5: 129-132.

6. Barbour T; Ramsden CT. 1916. Catalogo de los reptiles y anfibios de la Isla de Cuba. Memorias de la Sociedad

Cubana de Historia Natural “Felipe Poey”’ Habana 2: 124-143.

7. Barbour T; Ramsden CT. 1919. The herpetology of Cuba. Memoirs of the Museum of Comparative Zoélogy at

Harvard College 47: 69-213.

8. Barquero MD; Campos-Chinchilla P; Valverde R. 2005. Tretanorhinus nigroluteus: biology and captive maintenance

of the Orangebelly Swamp Snake. Reptilia 42: 47-52.

9. Bocourt MF. 1891. Sur quelques ophidiens de |’Amerique intertropicale: appartenant au genre 7retanorhinus. Le

Naturaliste 101: 121-122.

10. Boulenger GA. 1893. Catalogue of the Snakes in the British Museum (Natural History). Volume I. Taylor and

Francis, London, United Kingdom. 504 p.

11. Boulenger GA. 1903. Descriptions of new snakes in the collection of the British Museum. Annals and Magazine of

Amphib. Reptile Conserv. 234 December 2019 | Volume 13 | Number 2 | e208

12,

13.

4].

42.

Barquero and Arguedas

Natural History: Zoology, Botany, and Geology Series 7, 12: 350-354.

Cadle JE. 1984. Molecular systematics of Neotropical xenodontine snakes: H. Central American xenodontines.

Herpetologica 40: 21-30.

Cadle JE. 1985. The Neotropical colubrid snake fauna (Serpentes: Colubridae): Lineage components and

biogeography. Systematic Zoology 34: 1—20.

. Campbell JA. 1999. Amphibians and Reptiles of Northern Guatemala, the Yucatan, and Belize. University of

Oklahoma Press, Norman, Oklahoma, USA. 400 p.

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Amphib. Reptile Conserv. 237 December 2019 | Volume 13 | Number 2 | e208

Appendix 2. Number of specimens of 7retanorhinus held in each scientific collection from which data were obtained.

Specimens Abbrev

7 ANSP

143 AMNH

1 BYU

59 CAS

12 CM

8, CHP

12 FMNH

82 FLMNH

6 FHGO

12 INHS

5 IBUNAM

8 IAvH

35 LACM

64 LSUMZ

45 MPM

15 MNHN

60 MCZ

3 MVZ

By USNM

1] BMNH

5 RBINS

3 ROM

3 TCWC

vs MHUA

12 MZUCR

2 UVC

1 UTCH

2 UCM

77 KU

10 UMMZ

4 UTA

7355 Total

Amphib. Reptile Conserv.

Biology of Tretanorhinus species

Institution name

Academy of Natural Sciences, Philadelphia

American Museum of Natural History

Bingham Young University, Monte L. Bean Life Science Museum

California Academy of Sciences

Carnegie Museum of Natural History

Circulo Herpetologico de Panama

Field Museum of Natural History

Florida Museum of Natural History

Fundacion Herpetologica Gustavo Orcés

Illinois Natural History Survey, University of Illinois (formerly University of

Illinois Museum of Natural History [UIMNH])

Instituto de Biologia, Universidad Nacional Autonoma de México

Instituto de Investigacion de Recursos Biologicos Alexander von Humboldt

Los Angeles County Museum of Natural History

Louisiana Museum of Natural History (formerly Louisiana State University,

Museum of Zoology)

Milwaukee Public Museum

Muséum National d’ Histoire Naturelle

Museum of Comparative Zoology, Harvard University

Museum of Vertebrate Zoology, University of California at Berkeley

National Museum of Natural History, Smithsonian Institution (formerly United

States National Museum)

Natural History Museum (formerly British Museum of Natural History)

Royal Belgian Institute of Natural Sciences

Royal Ontario Museum

Texas Cooperative Wildlife Collection

Universidad de Antioquia, Museo de Herpetologia

Universidad de Costa Rica, Museo de Zoologia

Universidad del Valle

Universidad Tecnologica del Choco

University of Colorado, Museum of Natural History

University of Kansas, Biodiversity Institute

University of Michigan, Museum of Zoology

University of Texas at Arlington

238 December 2019 | Volume 13 | Number 2 | e208

Official journal website:

amphibian-reptile-conservation.org

Amphibian & Reptile Conservation

13(2) [General Section]: 239-258 (e209).

urn:lsid:zoobank.org:pub:7751CF14-97D4-4874-88CE-5567ED7B72AF

Three new species of Hemidactylus Oken, 1817

(Squamata, Gekkonidae) from Iran

'2:3Farhang Torki

'Razi Drug Research Center, Iran University of Medical Sciences, Tehran, IRAN *FTEHCR (Farhang Torki Ecology and Herpetology Center for

Research), 68319-16589, P. O. Box: 68315-139 Nourabad City, Lorestan Province, IRAN *Biomatical Center for Researches (BMCR), Khalifa,

Nourabad, Lorestan, IRAN

Abstract.—Based on morphological characters, three new species of the genus Hemidactylus are described,

one from the Zagros Mountains (Khuzestan Province) and two from the coastal Persian Gulf (Bushehr Province)

of Iran. The three new species can be differentiated from all other Hemidactylus inhabitants of Iran and adjacent

area congeners by distinct morphometric, meristic, and color characters. Comparisons with other species of

Hemidactylus are presented and a key to the genus is provided. Some information about the ecology, biology,

and conservation of the three new species is provided. Existing data suggest these geckos are point endemics.

Some additional historical information about the Hemidactylus inhabitants of Iran is discussed, particularly H.

parkeri.

Keywords. Bushehr, Hemidactylus achaemenidicus sp.n., H. pseudoromeshkanicus sp.n., H. sassanidianus sp.n., H.

parkeri, Khuzestan, Sauria, Reptilia

Citation: Torki F. 2019. Three new species of Hemidactylus Oken, 1817 (Squamata, Gekkonidae) from Iran. Amphibian & Reptile Conservation 13(2)

[General Section]: 239-258 (e209).

ternational (CC BY 4.0): https://creativecommons.org/licenses/by/4.0/], which permits unrestricted use, distribution, and reproduction in any medium,

provided the original author and source are credited. The official and authorized publication credit sources, which will be duly enforced, are as follows:

official journal title Amphibian & Reptile Conservation; official journal website: amphibian-reptile-conservation.org.

Received: 4 February 2019; Accepted: 20 June 2019; Published: 14 December 2019

Introduction

Globally, the gekkonid genus Hemidactylus Oken, 1817

currently consists of 154 species distributed across all

tropical and subtropical continental landmasses, including

intervening oceanic and continental islands (Carranza

and Arnold 2012; Smid et al. 2013a,b, 2015; Uetz 2019).

Four families and 70 species of geckos occur in Iran:

50 species of Gekkonidae, 10 species Phyllodactylidae,

seven species of Sphaerodactylidae, and three species

of Eublepharidae (Uetz 2019). The four Hemidactylus

species reported so far from Iran are: H. flaviviridis, H.

persicus, H. robustus, and H. romeshkanicus (Anderson

1999; Bauer et al. 2006a; Rastegar-Pouyani et al. 2006;

Torki et al. 2011; Kamali 2013; Smid et al. 2014). Only

one of them is endemic to Iran (H. romeshkanicus).

During a 2007-2010 collection program in south-

western Iran, from the Zagros Mountains and coastal

Persian Gulf, several geckos were collected which, upon

laboratory examination, were found to differ in important

characters from Iranian geckos already known.

In this article, they are described morphologically

and compared to the previously known Hemidactylus

species from Iran, as well as those from neighboring

Correspondence. torki,f@iums.ac.ir, torkifarhang@yahoo.com

Amphib. Reptile Conserv.

regions. Additionally, two notes regarding H. parkeri are

presented, and comments on the conservation of geckos

in Iran are provided.

Materials and Methods

During several field trips in the Iranian plateau, three

new Hemidactylus species were collected from this

region (Fig. 1): (a) Kangan region, near the coastal

Persian Gulf, Bushehr province, (b) Tangestan region,

Bushehr province, and (c) Kole-Saat, Khuzestan

province. All specimens of the three new species were

assigned catalog numbers for the ZFMK (Zoologisches

Forschungsmuseum Alexander Koenig, Bonn,

Germany); and FTHM (Farhang Torki Herpetological

Museum, Nourabad City, Iran), with the latter deposited

in Farhang Torki Ecology and Herpetology Center for

Research (FTEHCR).

The taxonomic characters of Hemidactylus species

from Iran are not well defined. For most species, no

museum specimens were available for comparison.

Rather, published descriptions of geckos known from

Iran were compared to the morphological characters of

the newly collected material (e.g., Moravec et al. 2011;

December 2019 | Volume 13 | Number 2 | e209

Three new Hemidactylus species from Iran

Andimeshk

Fig. 1. Type localities of three new geckos in Iran.

Carranza and Amold 2012; Smid et al. 2013a, 2015).

For comparison with H. romeshkanicus ZMB 75020

from the Museum fiir Naturkunde, Leibniz Institut fur

Biodiversitats- und Evolutionsforschung zu _ Berlin

(formerly Zoologisches Museum Berlin, Germany) was

used. For comparison with Hemidactylus spp. distributed

outside Iran, original descriptions or other publications

containing morphological analyses of Hemidactylus

species were used (e.g., Anderson 1999; Giri et al. 2003;

Baha el Din 2003, 2005; Bauer et al. 2006a,b, 2007;

Sindaco et al. 2007; Giri and Bauer 2008; Giri 2008;

Mahony 2010; Agarwal et al. 2011; Busais and Joger

2011; Moravec et al. 2011; Torki et al. 2011; Mirza and

Rajesh 2014; Vasconcelos and Carranza 2014; Carranza

and Arnold 2012; Smid et al. 2013a, 2015; Safaei-

Mahroo et al. 2017).

Characters were selected to optimize comparisons

with data reported by Moravec et al. (2011), Carranza

and Arnold (2012), Wagner et al. (2014), Vasconcelos

and Carranza (2014), and Smid et al. (2013a, 2015).

Measurements were taken using a dial caliper with 0.01

mm precision. Additionally, other characters important

for the taxonomy of Hemidactylus were used, such

as nasals in contact and 1“ postmental in contact with

2™ lower labial (e.g., Moravec et al. 2011; Smid et

al. 2013). Characters used to describe the three new

Hemidactylus are as follows: SVL: snout-vent length;

TRL: trunk length; TL: tail length; Rl: TL/SVL; HL:

head length; HW: head width; HH: head height; R2: HL/

Amphib. Reptile Conserv.

(H. pseudoromeshkanicus sp.n.)

, lange stan (H. sassanidianus sp.n.)

Kangan (H. achaemenidicus sp.n.)

SVL; R3: HW/HL; R4: HH/HL; OD: orbital diameter;

NE: nares to eye distance; IN: internarial distance; IOI:

anterior interorbital distance; [02: posterior interorbital

distance; TB: longitudinal tubercle rows; PAP: number

of precloacal pores; SL (L/R): number of supralabials; IL

(L/R): number of infralabial scales; LP1 (L/R): number

of lamellae under the first finger of the pes; LP4 (L/R):

number of lamellae under the fourth finger of the pes; FP:

femoral pores; and PM: postmentals. Abbreviations used

in tables are as follows: M: male; F: female; T: total; A:

ANOVA test; F: one-way ANOVA F value; dF: degrees

of freedom; P: probability; DM: Difference of means;

and DD: Direction of difference.

Because of the absence of sexual size dimorphism

in the arid clade of Hemidactylus (Carranza and Arnold

2012), both sexes were analyzed together. Statistical

procedures used to test for differences between the sexes

included one-way ANOVA (at 95% confidence level [P <

0.05]) and Principal Component Analysis (PCA).

Taxonomy

Hemidactylus achaemenidicus sp.n. (Figs. 2—5)

Hemidactylus turcicus - Torki et al. 2011

Hemidactylus persicus - Carranza and Arnold 2012

Hemidactylus persicus - Smid et al. 2013

urn: sid: zoobank.org:act:40139EE0-898A-4E3B-B9B7-32C73 FE 16377

December 2019 | Volume 13 | Number 2 | e209

Fig. 2. Dorsal and ventral views of (a, b) holotype and (ce, d)

paratype specimens of Hemidactylus achaemenidicus sp.n.

Holotype

ZFMK 98567, adult male, collected at the end of the southern

Zagros Mountains, Kangan, Bushehr Province, Southern

Tran, on 10 May 2008 (27°18’N, 52°42’E, 50-221 m asl).

Paratypes

ZFMK 97750-97753; ZFMK 98568—-73; and FTHM

005110, six adult male specimens (ZFMK 97750-97752;

ZFMK 98569-—70; FTHM 005110), and four adult female

specimens (ZFMK 97753; ZFMK 98568, 71, 72), same

data as for holotype.

Diagnosis

A small sized Hemidactylus, maximum snout-vent

length 39.8 mm; tubercles distributed over the entire

dorsum (except for forelimbs); granules cover head and

extend to neck; tubercle rugosity dimorphism occurs

between males and females over dorsal body, limbs, and

tail (males have more rugose tubercles than females):

proximal portion of tail (ventral view) covered by small

scales without femoral pores; precloacal pores present;

six tubercles on most whorls of tail; two postmentals; low

number of lamellae under pes; subcaudal scales started

more distally (approximately after proximal one-third of

tail), only a few subcaudals (plate-like) in original tail

(O—22), that started so far as anal; proximal dorsal tail

covered by regular whorls of tubercles (keeled in male

and plate-like in female); ventral scales not imbricate; the

ends of ventral scales are denticulated; enlarged scansors

beneath fingers, scansors are mostly divided, terminal

scansor is single; dorsal color pattern shows much

variability (regular or irregular crossbars, longitudinal

bands, large or small spots), and this is true for the tail

(regular or irregular bars, large and small spots), venters

of all specimens are without spots (uniform).

Description of Holotype (Figs. 2—3)

Measurements (in mm): body size: 39.8; tail length: 40.5;

interlimbs: 18.3; head width: 7.3; head length: 11.7; head

depth: 4.9; eye-eye: 4.7; ear opening: 0.82; eye diameter:

3.0; forelimb length: 12.3; hindlimb length: 15.5.

Amphib. Reptile Conserv.

Fig. 3. (a) Postmentals and (b) precloacal pores in holotype of

Hemidactylus achaemenidicus sp.n.

Body depressed, tail more or less flattened; head

triangular-shaped; two postmentals, 1 postmentals

enlarged and in contact, 2" postmentals behind the first

enlarged postmentals, the 1“ postmentals in contact

with the 1% infralabials, the 2" left postmental distinct

from infralabials by one series of scales, the 2" right

postmentals in contact with the 2™ infralabials (and

weakly with the 1*), four scales between 2™ postmentals;

Infralabials: eight; supralabials: nine; nostril surrounded

by five scales (the 1“ supralabial, rostral, three small on

posterior); nasals not in contact and separated by one

scale; ear openings more or less falcate-shaped, and

horizontal; 14 scales between nostril and eye; 24 scales

between eye and ear; rostrum covered by large granules;

space between eyes covered by 27 small granules, and

10 small simple tubercles distributed among them; upper

head covered by smallest granules and many small simple

tubercles distributed among them; tubercles on upper ears

and behind eyes are simple; tubercles on occiput mostly

simple and less pointed; tubercles on neck are pointed

and keeled (heterogeneous); from rostrum to neck body

covered by granules; tubercles distributed on dorsum,

head, and limbs; tubercles not found on arm; most body

tubercles are keeled; dorsal tubercles are strongly keeled,

December 2019 | Volume 13 | Number 2 | e209

Three new Hemidactylus species from Iran

Fig. 4. Subcaudal of tail of (a) ZFMK97753 and (b) ZFMK 98567 of He

between (c) holotype of H. achaemenidicus sp.n. and (d) lectotype of H. robustus. Photo from Smid et al. 2015.

tubercles on proximal of back surrounded by 11-12

scales (middle: 12-13, distal: 13), dorsal tubercles do

not show regular form and abnormalities occur in a few

points (intermixed with some small simple tubercles);

enlarged, trihedral, and strongly keeled tubercles

distributed on distal part of dorsum (between hindlimbs)

as well as nearest to tail; tubercles on forearm are simple;

tubercles on femur heterogeneous (simple, pointed, and

keeled); foreleg tubercles heterogeneous in size and

shape (pointed and keeled); size of the tubercles on limbs

is different and is as follows: foreleg > femur > forearm;

scales on palm and sole are granule-like; 17 rows (mostly

regular) of tubercles on back; 21 tubercles between

interlimbs.

Tail is original; first part of tail (one-third) covered

by small scales, subcaudal plates cover following third,

less than 12 scales (moderate size: 50% of tail width, not

imbricate) on subcaudal, last part; distal one-third of tail

is without subcaudals (covered by small scales); without

crossbars on dorsum of tail, small irregular spots present

in first half of tail; tubercle whorls only found on first

half of tail, 1‘ to 6" whorls more or less irregular and

separated by one scale, includes six large, trihedral, and

strongly keeled tubercles, after them real whorls start:

six tubercles in 1* to 3 whorl, four for 4" to 7", first

whorl separated from secondary by two scales, four

scales between 2"*-3" and 34". six scales between 4°—

5". five between 5-6" and 6"—7"| after them tubercles

converted to scales; seven (3+1+3) precloacal pores; no

femoral pores; enlarged scansors are plate-like; terminal

scansor is single; lamellae on fingers as follows: 1*: five,

2": seven, 3™: seven, 4": seven, 5": eight; lamellae on

pes as follows: 1%: six, 2": eight, 3: nine, 4": 10, 5%:

eight; claws in front of scansors. Palm and sole covered

by granule-like scales.

Coloration of upper head is covered by longitudinal

discontinuous rows that extended to neck, and are

Amphib. Reptile Conserv.

irregular onto dorsal body; dorsum without bars; few

irregular bars and small spots cover dorsum of tail;

one bar between nostril-eye-ear; venter of body, limbs,

and tail uniformly without pattern; pattern in preserved

specimen is similar to the live specimen and all spots and

bars are obvious; the preserved specimen is colorless.

Variation (Fig. 4a—b)

Some variation among paratypes 1s described as follows:

tubercles distributed all over dorsum (except arms);

granules cover head and extend to neck. Tubercle

rugosity differed between males and females on overall

dorsal body, limbs, and tail (males with more strongly

rugose tubercles than females), females have wide

(approximately flattened shape) dorsal tubercles and

males have extended trihedral tubercles. Proximal tail

in most specimens is cycloid and ventral view covered

by small scales (same as dorsum); proximal tail (dorsal

view) covered by 4-6 irregular whorls of tubercles

(strongly keeled) separated by one scale, followed by

regular whorls of six tubercles in each whorl, started and

separated by 2-6 scales, more than six regular whorls are

obvious in all specimens (first half) and do not continue

to posterior half of tail (tubercles converted to scales).

Number of precloacal pores is variable as follows: six (five

Specimens), seven (holotype), and eight (ZFMK97751).

Most specimens have two postmentals, postmentals in all

specimens are not uniform and variability is as follows:

one specimen (ZFMK 98570) has five postmentals

(left+right), two anterior, two posterior, and one large

scale between anteriors; anteriors not in contact with

one another and in contact with 1% and 2" infralabials;

seven specimens have normal postmentals and anterior

in all specimens in contact with 1* and 2" infralabials; in

one specimen (FTHM005110) 2" postmentals (left and

right) separated from infralabials by one series of scales,

and 1‘ postmentals in contact with 1* infralabial; left 2"

December 2019 | Volume 13 | Number 2 | e209

midactylus achaemenidicus sp.n. Comparison of dorsal body

Torki

Fig. 5. Type locality of Hemidactylus achaemenidicus sp.n.,

Kangan, Bushehr, southern Iran.

postmental in ZFMK97751 separated from infralabials

by one series of scales, and 1‘ postmentals are in contact

with 1* infralabials; finally left 1‘ postmental of ZFMK

98571 contacts 1* infralabials. Subcaudal scales begin

approximately after first third of tail, a lesser number

of subcaudals (plate like) in original tail (0 to 22); first

half of dorsal tail covered by regular whorls of tubercles

(strongly keeled in males and plate-like in females).

Dorsal color pattern is variable (regular or irregular cross

bars, longitudinal band, large or small spots), this is true

for tail (regular or irregular bars, large and small spots),

venter of all specimens is uniform, without spots; venter

in live specimens is white and tail is yellowish or dark,

in preserved specimens ventral is yellowish and ventral

of tail is darkish. More data on the variation are shown

in Table 1.

Sexual dimorphism is evident. In general, males show

larger body size and head size than females (Table 1).

Amphib. Reptile Conserv.

Based on statistical analysis three characters, TRL, [O2,

and LPAR, are significantly different between the sexes

as follows: males have significantly (P = 0.03, f = 6.21)

larger trunk length than females (16.9 + 0.37 vs. 14.3 +

1.28); this is true for IO2 and in males (4.70 + 0.1) is

significantly (P = 0.03, f = 6.09) larger than in females

(4.22 + 0.18). In contrast, number of lamellae under 4"

pes (right side) in females (10 + 0.0) is significantly (P

= 0.01, f = 8.18) greater than in males (9.28 + 0.18).

Five characters (SVL, TRL, TL, HW, HL) in females

show much more variability than in males; in contrast,

three characters (OD, NE, IN) in males are much more

variable than in females. All females have 16 dorsal

tubercle rows, and in males they number 16 or 17 (16.4 +

0.2). Lamellar variability under 1‘ and 4" finger of pes in

females is zero and in males is one (except ILL: female

is one and males are zero). More data on the dimorphism

are shown in Table 1.

Habitat and Ecology (Fig. 5)

Hemidactylus achaemenidicus sp.n. are distributed in the

eastern part of Bushehr Province (edge of Hormozgan

Province), in Kangan, Assaloye City. The habitat of H.

achaemenidicus sp.n. is flat land covered by Jujube trees

(Ziziphus jujuba). The type locality is located in the

northern part of the Persian Gulf. A few lizard and snake

Species were observed at the type locality: 7rapelus

agilis, Laudakia nupta, and Echis carinatus.

Distribution

So far, the species is only known from the type locality.

Etymology

The species name “achaemenidicus” refers to “The

Achaemenid Empire,” also called the First Persian

Empire. It was an empire based in Western Asia, founded

by Cyrus the Great, and notable for including various

civilizations and becoming the largest empire at that

time.

Comparisons

Based on a phylogenetic study of one paratype specimen

(FTHM 005100 is erroneous and FTHM 005110 is the

true code; also the locality cited in the phylogeny section

must be changed to the type locality of the new species)

H. achaemenidicus sp.n. is completely distinct from

H. robustus, H. turcicus, and other recently described

species inhabiting Oman (see phylogram of Carranza

and Arnold 2012; Smid et al. 2015). Hemidactylus

achaemenidicus sp.n. was compared with the re-

description of H. robustus Smid et al. (2015) [see Table

2]. Hemidactylus achaemenidicus sp.n. is different from

H. robustus by smaller body size in males (36.5 + 0.9 mm

vs. 41.8 + 2.3) and females (33.1 + 2.0 mm vs. 43.6 +

4.7), more longitudinal tubercle rows (16.2 + 0.1 vs. 14.8

+ 1.2), and keeled (vs. weakly keeled and posteriorly

pointed) as well as rugosity dimorphism (quite distinct

December 2019 | Volume 13 | Number 2 | e209

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December 2019 | Volume 13 | Number 2 | e209

244

Amphib. Reptile Conserv.

Table 2. Comparison three new Hemidactylus species with other Hemidactylus which occur in Iran. Data from: (1): Carranza and Arnold

Torki

2012; (2): Torki et al. 2011; (3): FTHM collections. Abbreviation are as given in Materials and Methods and Table 1 header.

Characters § H. achaemenidicus sp.n. H. sassanidianus sp.n. —_H.. pseudoromeshkanicus spn. _—_-H.. persicus (1) H. romeshkanicus (2) H. flaviviridis (3)

SVL 28-39 48-63 74-75 36-67 70 59-79

TL 24-41 66-79 88 55-77 83 60-97

SL 9-10 10-13 11 10-13 15 12-17

IL 7-8 8-10 9 8-11 9 11-12

HL 8.7-12.2 13.5-20.6 22.8-23.4 9.1-16.8 23.2 17-23

HW 5.4-7.5 9.7-13 14.6-15.2 7.1-14.4 14.5 12.5-18.5

HH 3.7-5.3 6.1-8.9 8.7-9.1 4.9-9.6 9.1 7.9-10.9

HL/SVL 0.27-0.31 0.28-0.31 0.30-0.31 0.21-0.28 0.33 0.28-0.31

HW/HL 0.60—-0.68 0.60-0.71 0.64—0.65 0.67-0.92 0.62 0.65-0.80

HH/HL 0.39-0.47 0.35-0.45 0.37-0.40 0.42-0.60 0.39 0.40-0.48

DTR 16-17 14-16 16-17 14-16 16 -

PAP 6-8 6-8 12 8-11 12 -

LPI 5-6 6-9 11 8-9 8 8-9

LP4 9-10 9-14 15 13-14 12 11-13

PM 2 2-4 2 2 3 2

for new Hemidactylus species), subcaudal scales (scale

like and/or enlarged vs. enlarged), less head width/head

length (0.62 vs. 0.74), internarial distance (0.97 + 0.04

vs. 1.5 + 0.08), lower number of lamellae under the 1“

pes (5.7 + 0.1 vs. 6.1 + 0.5), internarial distance (0.97 vs.

1.5), and nasal in contact % (0% vs. 22%) [Carranza and

Arnold 2012; Smid et al. 2015]. Based on photograph of

lectotype of H. robustus (Figs. 4-9 in Smid et al. 2015;

as a female specimen), females of H. robustus have

approximately full rugosity (lectotype of H. robustus is

female) and it is more than in male H. achaemenidicus

sp.n. (males of H. achaemenidicus sp.n. have much

greater rugosity than females); dorsal tubercle density

(especially on proximal part) in H. robustus 1s more than

H. achaemenidicus sp.n. dorsal, and dorsolaterals of

H. robustus have maximum uniformity; in contrast the

dorsum of H. achaemenidicus sp.n. has heterogeneity

of dorsal and dorsolateral tubercles; also shape and size

of tubercles on dorsolateral of H. achaemenidicus sp.n.

is different from mid-dorsum, in contrast to H. robustus

(Fig. 4c—d); photographic comparison: limbs (especially

hind limbs) in H. achaemenidicus sp.n. are smaller than

H. robustus, additional differences are: longer head for

H. robustus; smaller interlimbs for H. robustus, base

of tail in H. robustus is much more flattened and in H.

achaemenidicus sp.n. is approximately cylindrical.

Differs from H. flaviviridis, H. persicus, and H.

romeshkanicus by smaller body size. More comparisons

with Hemidactylus inhabiting Iran are shown in Table 2.

Differs from H. turcicus by smaller body size (36.5 + 0.9

mm vs. 46.0 + 5.8 in males, 33.1 + 2.0 mm vs. 49.2 +

5.1 in females), short tail relative to SVL (TL 0.98 vs.

112.8% of SVL), more longitudinal tubercle rows (16.2

+ 0.1 vs. 13.8 + 0.7), nasal in contact % (0% vs. 13.3%),

1s' and 2" postmentals in contact with 2" infralabials

(81.8% vs. 12.9%), lower number of lamellae under the

1* pes (5.7 vs. 6.6), supralabials (9.5 + 0.1 vs. 8.3 + 0.5),

infralabials (7.8 + 0.1 vs. 6.8 + 0.4), number of precloacal

pores (6.42 vs. 7.2), less head width/head length (0.62 vs.

Amphib. Reptile Conserv.

0.77) [Moravec et al. 2011; Smid et al. 2013]. Different

from H. persicus in body size, tail length, head shape

and ratio, dorsal tubercle rows, precloacal pores, and

number of lamellae under the 1% and 4" pes (see Table

2). Different from H. romeshkanicus in body size, tail

length, head shape, precloacal pores, and number of

lamellae under the 1‘t and 4" pes (see Table 2).

In this section H. achaemenidicus sp.n. is briefly

compared with other Hemidactylus spp. from Iran. Different

from H. adensis, H. awashensis, H. lavadeserticus, H.

mandebensis, H. ulii, and H. jumailiae by more longitudinal

tubercle rows (16.27 vs. 14, 14, 14, 13.3, 14.1, and 14)

[Smid et al. 2013a, 2015]. Different from H. dawudazraqi

by more dorsal tubercle rows (16-17 vs. 12-15). Different

from H. alfarraji by precloacal pores (6-8 vs. 4) [Smid et

al. 2016]. Different from H. kurdicus by postmentals (2 vs.

1) [Safaei-Mahroo et al. 2017]. Different from H. foudaii

by precloacal pores (6-8 vs. 9) and well developed dorsal

and tail tubercles (vs. less developed and protuberant

dorsal and particularly tail tubercles). Different from

H. mindiae (Jordan) and H. asirensis by smaller body

size (36.5 mm vs. 49.3, 43-48.5 in males, 33.1 mm vs.

49.8, 38-51 in females, respectively) [Baha el Din 2005,

Moravec et al. 2011; Smid et al 2017]. Different from H.

saba, H. granosus, H. yerburii, H. montanus, H. minutus,

H. homoeolepis, and H. mindiae (Egypt population) by

number of precloacal pores (6.42 vs. 8, 5.6, 13.7, 11.2, 5.8,

4.3, 12.8, and 4) [Baha el Din 2005; Carranza and Arnold

2012: Smid et al. 2013a, 2016; Vasconcelos and Carranza

2014], respectively. Different from H. endophis by

lacking femoral pores. Different from H. shihraensis, H.

hajarensis, H. luqueorum, H. festivus, and H. alkiyumii by

smaller body size. Significantly different from H. mindiae,

H. lavadeserticus, H. dawudazragi, H. shugraensis, and

H. sinaitus by small body size and more dorsal tubercle

rows. Different from H. leschenaultii, H. homoeolepis,

H.. paucituberculatus, H. inexpectatus, H. masirahensis,

and H. /emurinus by having large and keeled tubercles on

dorsal body.

December 2019 | Volume 13 | Number 2 | e209

Three new Hemidactylus species from Iran

Fig. 6. Dorsal tubercles of (a) holotype and (b) paratype of

Hemidactylus sassanidianus sp.n.

Based on recent a molecular study on Hemidactylus

(Maximum-likelihood tree inferred using 350 bp of

the 12S gene, Appendix HI, by Carranza and Arnold

2012; Smid et al. 2013b, 2015), H. achaemenidicus

sp.n. (FTHMO005110 is the accurate specimen number)

is significantly different from: H. /uqueorum, H.

hajarensis, H. lemurinus, H. yerburii, H. montanus,

H. jumailiae, H. alkiyumii, H. robustus, H. sinaitus, H.

saba, H. shihraensis, H. festivus, H. paucituberculatus,

H. masirahensis, H. inexpectatus, and H. homoeolepis.

Hemidactylus sassanidianus sp.n. (Figs. 6—9)

Hemidactylus persicus Torki et al. (2011)

urn:Isid:zoobank.org:act:61 CDBB8A-CE1 F-4219-8F66-9DB6275C577E

Holotype

ZFMK 98573, adult male, collected at the southern end

of Zagros Mountains, Khaiiz, Tangestan City, Bushehr

Province, Southern Iran, on 4 May 2008 (28°43’N,

51°31’E, 525 m asl).

Paratypes

ZFMK 97754—-56, ZFMK 98574—77, FTHM 005029;

four adult male specimens (ZFMK 97756, ZFMK

98575-77), and four adult female specimens (ZFMK

Amphib. Reptile Conserv.

97754—55, ZFMK 98574, FTHM 005029), same data as

for holotype.

Diagnosis

A small-sized Hemidactylus, snout-vent length at

least 48.3 mm; tubercles distributed all over dorsum,

except for arm; back with enlarged keeled tubercles;

heterogeneity of dorsal tubercles occurred in all

specimens (a few parts or most of dorsal body);

dorsal scales in a few places converted into granules;

granules cover snout, between eyes, upper head, neck,

and in some specimens onto middle of dorsum and

dorsolaterals; 2—4 postmentals; 4-8 whorls of tubercles

on first half of dorsum of tail, distal part of tail without

tubercles; without femoral pores; precloacal pores

present; more lamellae under fingers; subcaudal

scales enlarged; ventral scales not imbricate; enlarged

scansors beneath fingers, scansors mostly divided,

terminal scansor single; limbs without color pattern

and uniform, dorsolaterals without any pattern and

uniform, pattern only present on middle part of dorsum

(longitudinal) of all specimens, various patterns on

dorsum such as: spotty (small or large), bars (irregular

and regular); ventrum without pattern.

Description of Holotype (Fig. 6)

Measurements (in mm): body size: 54.2; tail length:

79.3; interlimbs: 21.6; head width: 10.6; head length:

16.4; head depth: 6.4; eye-eye: 6.2; ear opening: 1.9; eye

diameter: 4.3; forelimbs length: 18.3; hind limbs length:

24.8.

Body depressed; body, as well head are flattened; tail

flattened; head triangular-shaped; two postmentals, the

first postmentals are enlarged and are widely in contact

together, the 2"! postmentals one behind the first enlarged

postmentals, the 1‘ postmentals are in contact with the

1st infralabials, the 2™ postmentals are in contact with

the 2"! infralabials, four scales between 2™ postmentals;

infralabials: nine; supralabials: left: 11, right: 12; nostril

surrounded by five scales (the 1% supralabial, rostral,

internasal scale and two postlabials); nasals not in contact

and separated by one small scale; ear openings are falcate-

shaped, and horizontal; 14 scales between nostril and eye;

26 scales between eye and ear; 31 scales between eyes;

rostrum covered by large granules and a few tubercles

distributed in distal part; between eyes covered by small

granules, and nine small smooth and simple tubercles

distributed among them; upper head covered by smallest

granules and small tubercles distributed among them;

tubercles on upper ears simple and pointed; tubercles

on occipital are mostly pointed; tubercles on neck are

simple, pointed and keeled (heterogeneous); granules

cover rostrum to neck body; tubercles distributed on

dorsum, head, and limbs; tubercles extend to in front of

eyes; tubercles not found on arm; most body tubercles are

keeled; dorsal tubercles are keeled, a few areas of mid-

dorsum covered by abnormal tubercles (heterogeneous in

December 2019 | Volume 13 | Number 2 | e209

Torki

Fig. Te Postmental variation in Hemidactylus SETTER

sp.n. (a) ZFMK 98573; (b) FTHM 005029; (c) ZFMK 97756;

(d) ZFMK 98575.

shape and type) and granules; dorsolateral tubercles are

keeled and wide; forearm tubercles are small and simple;

size of the forearm tubercles are smaller than hindlimb

tubercles; number of tubercles on femur (pointed and

keeled) are less than foreleg (mostly keeled); scales on

palm and sole are granular; 16 regular rows of tubercles

on back; 6—8 small simple tubercles between interorbits,

32 scales between interorbits (mid-part); 22 enlarged

tubercles between fore- and hindlimbs; 12-14 scales

surround each mid-dorsal tubercle (11-12 proximally,

12-13 distally); 3-4 scales between each dorsal tubercle.

Tail is original; 52 enlarged imbricate subcaudal

scales; last part of tail cycloid-shape and covered

by raised scales; proximal of tail covered by several

continuous indistinct bars, 13 crossbars on dorsum of

tail, tubercle whorls only found in anterior part of tail,

5—6 scales between each whorl, six tubercles in first

whorl, six tubercles in the second, six in third, five in

fourth, five in fifth, six in sixth, and six tubercles in the

seventh whorl, after the seventh whorl tubercles become

very small (six in each whorl) and converted into scales;

ventral scales (mostly oval shape) are not imbricate and

their size in the middle part of the body are larger than

other regions; eight (4+4) precloacal pores; without

femoral pores; enlarged scansors are plate-like, terminal

scansor 1S unique (not paired); lamellae on fingers as

follows: 1‘: nine (1-3 undivided), 2": 10 (1 undivided),

3 10 (1 undivided), 4: 11, 5%: 11(1-2 undivided);

lamellae on pes as follows: 1%: nine (1-2 undivided),

2™: 11 (1 undivided), 3": 12 (1 undivided), 4": 14 (1-2

undivided), 5": 13; palm and sole covered by granule-

like scales.

Coloration: upper head, neck, and middle part of

dorsum covered by smallest spots and few large paled

spots (background view), don’t form bar; without spots

or bars on dorsolaterals and limbs; one narrow stripe

between nostril-eye and eye-ear; three moderate spots

on snout; a paled and irregular bar on occipital and neck;

Amphib. Reptile Conserv.

he y f ‘iit “Y J

ey wot 9 Le ee #35 is

ety ‘ } ie 2, tp ry a

Fig. 8. (a) Precloacal pores (ZFMK 98573), and (b) dorsal

tubercles (ZFMK 98573) of Hemidactylus sassanidianus sp.n.

color of venter is uniform white; palm of digits (hindlimbs

and forelimbs) more or less white; pattern of preserved

Specimen is similar to the live specimen, but has lost color.

Variation (Figs. 7-8)

Heterogeneity of dorsal tubercles occurs in a few areas,

mostly on the dorsal body; tubercles converted to simple

(not-keeled) and have abnormal shape (e.g., rounded,

width, semi), in these parts most dorsal scales converted to

granules (small or large); granules cover snout and extend

to upper head and neck (all specimens), or onto proximal

dorsum (ZFMK 97754 and 28) or onto mid-dorsum

(ZFMK 97755), lateral sides of neck strongly covered by

granules and tubercles (ZFMK 97756); most specimens

have 1—2 tubercles in front of ear, or 4-5 (ZFMK 98573

and ZFMK 97756) or lack tubercles (ZFMK 98577);

internasals in four specimens are in contact (ZFMK

98573, ZFMK 98575, ZFMK 97754—55) and in others

separated by one (ZFMK 97756, FTHM005029), two

(ZFMK 98577) or three (ZFMK 98574) scales; number

of postmentals is variable (usually two) between 2-4,

asymmetry occurs in some of them; ZFMK 98575 have

four PM as follows: 1‘ PM is large and in contact with

1st and 2™ infralabials, 2" PM on posterior of 1** PM and

in contact with 2™ infralabials, 3" PM behind 2"? PM

and separated from infralabials by one series of scales;

4‘t PM in contact with 1‘ and 2™ postmentals; ZFMK

97756 has three postmentals as follows: 1* larger and in

contact with 1* and 2™ infralabials, 2" PM is behind 1*

PM and in contact with 2" infralabials, 3" PM is behind

2™ PM and separated from infralabials by one series of

scales, 10 scales between 2" postmentals; FTHM005029

has three PM on left and two on right; ZFMK 98574

has three PM on left and two on right; 4-8 whorls of

tubercles on proximal half of dorsum of tail (usually

six), without tubercles on distal part of tail; limbs and

December 2019 | Volume 13 | Number 2 | e209

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December 2019 | Volume 13 | Number 2 | e209

oe)

Amphib. Reptile Conserv.

dorsolaterals are uniformly without pattern, pattern only

present on middle part of dorsum (longitudinal) of all

specimens, various patterns are visible on dorsum such

as: spots (small or large), bars (irregular and regular);

ZFMK 98574: 1% bar is full and wide, 2"! as well as 3%

are X-shaped, and usually 4" bar as well; ZFMK 98575:

narrow longitudinal stripe on middle part of dorsum,

two bars (usually X-shaped) on neck; FTHM005029:

six bars on dorsum, five regular and one irregular, one

irregular bar on neck; FTHM005029: one longitudinal

stripe from neck to tail; upper head and between eyes of

most specimens covered by small spots. More data on the

variations are shown in Table 3.

Sexual dimorphism is evident. In general, 12

characters are larger in females and 11 characters larger

in males. Females (26.5 + 0.74) have significantly (P =

0.01, f = 9.59) larger trunk length than males (22.8 +

0.87), as the result of fecundity selection (e.g., Andersson

1994; Torki 2012), and females have a larger trunk for

the development of two large eggs. The lamellae in

females (1%: 9; 4%: 13.7) number more than in males

(1s: 8; 4%: 12.2), which may be the result of natural

selection, because during development of the two large

eggs females must have more ability to move (Torki

2012); also, females have minimal variability of number

of lamellae (1%: 0; 4: 1) in contrast to males (1: 3; 4":

5); greater number lamellae and minimal variability in

females are important positive results of natural selection

for survival of H. sassanidianus sp.n. under natural

conditions (personal assumption of author). In general,

all characters (except dorsal tubercles rows), especially

body size (range: males: 15; females: 2.9) have more

variability in males. More data on the dimorphism and

variability are shown in Table 3.

Habitat and Ecology (Fig. 9)

The Tangestan region is at the end of the southern part of

the Zagros Mountains, and has palm trees. Hemidactylus

Sassanidianus sp.n. is distributed in a mountainous

area. This mountain is one of the Zagros Mountains

and its structure is sedimentary. Shelter sites of the

new Hemidactylus species are limited to the clefts and

Amphib. Reptile Conserv.

Fig. 9. Type locality of Hemidactylus sassanidianus Sp.n., Tangestan, Bushehr, southern Iran.

en.

meet

a :

gre

caves in this mountain, with many specimens found and

collected in one cave in this locality. This cave is deep—

the author was able to reach a depth of more than 50 m,

though the depth of this cave is said to be even more

than 200 m. This cave is an important habitat for this

new Hemidactylus and the largest population was seen

only in this cave. Other species of gecko were also seen

in this cave, such as Asaccus tangestanensis. Of the three

new species described here, only H. sassanidianus sp.n.

was seen in this cave, but H. sassanidianus sp.n. was not

seen outside of the cave or elsewhere in the entire region.

This cave probably opens into other regions, and further

investigation of this cave is needed. This cave is dark

during both day and night. Conditions inside the cave are

moist, in contrast to the conditions outside the cave.

Distribution

Hemidactylus sassanidianus sp.n. is distributed only

at the type locality, in Khaiiz, Tangestan City, Bushehr

Province, southern Iran. The type locality is situated at

the end of the southern Zagros Mountains, approximately

150 km from the Persian Gulf.

Sympatric Lizards and Snakes

Several lizard and snake species were observed in

the type locality, including Asaccus tangestanensis,

Laudakia nupta, Trapelus agilis, Tropiocolotes persicus,

Coluber (sensu lato) sp., Macrovipera lebetina, and

Echis carinatus.

Etymology

The species name “sassanidianus” refers to “The

Sasanian Empire,” also known as Sassanian, Sasanid,

Sassanid or Neo-Persian Empire, which was known to its

inhabitants as Eranshahr in the Middle Persian language.

Comparisons

Hemidactylus sassanidianus sp.n. differs from H.

persicus (based on original description by Anderon

1872) by: (1) Dorsal tubercles in H. sassanidianus sp.n.

are not strongly keeled and in some parts tubercles are

not keeled, in contrast they are strongly keeled in H.

December 2019 | Volume 13 | Number 2 | e209

Three new Hemidactylus species from Iran

Table 4. Sexual dimorphism among Hemidactylus persicus, H. sassanidianus sp.n., and H. achaemenidicus sp.n. Abbreviations:

HL: head length; HW: head width; HH: head height; IO1: anterior interorbital distance; [O02: posterior interorbital distance; SL:

number of supralabial; IL: number of infralabial scales (all data are means). F: female, M: male, sig: significant (Carranza and

AE:

rae ar

ae Ce

001

ee ee ce

0.29

Arnold 2012).

H. persicus

H. sassanidianus sp.n.

H. achaemenidicus sp.n.

persicus. (2) Heterogeneity of dorsal tubercles occurred

in all specimens of H. sassanidianus sp.n., in contrast to

original description of H. persicus. (3) Size of tubercles

in H. sassanidianus sp.n. is smaller than H. persicus,

about 0.4 of ear opening vs. 0.5 ear opening. (4) Five or

six tubercles in each row of the tail in H. sassanidianus

sp.n., and in contrast H. persicus have seven tubercles in

each row of the tail. (5) Dorsal body of H. sassanidianus

sp.n. covered by spots (not bars), in contrast dorsal body

in H. persicus is covered by transverse narrow band.

More differences between H. sassanidianus sp.n. and

H. persicus (based on original description and Anderson

1999): (6) H. persicus only has two postmentals (in all

populations; there are no records in the literature), in

contrast H. sassanidianus sp.n. has 2—4 postmentals.

(7) In Anderson’s work on H. persicus inhabiting Iran,

he reported 9-11 preanal pores, which is clearly more

than H. sassanidianus sp.n. (6-8). (8) Tail sharp in

H. sassanidianus sp.n., and not sharp in H. persicus.

Additional differences with H. persicus include:

number of postmentals (2-4 vs. 2), mental trihedral (vs.

pentagonal); relatively fewer precloacal pores in males

(6-8 vs. 9-11); number of lamellae under the first digit

of the pes (6—9 vs. 8-9); body size of H. sassanidianus

sp.n. males (54.7) smaller than females (56.4), this is in

contrast to H. persicus (males: 59; females: 51.4); head

longer (HL/SVL: 0.3 vs. 0.24), elongated (HW/HL:

0.64 vs. 0.8), and more flattened (HH/HL: 0.4 vs. 0.49)

[Carranza and Arnold 2012]; sexual dimorphism in head

size (HL, HW, and HD) occurs for H. persicus (males

significantly larger than females), this is in contrast to H.

sassanidianus sp.n., and this is true for more characters

(Table 4). Easily differentiated from H. romeshkanicus by

number of precloacal pores (6-8 vs. 12), other differences:

smaller body size, number of supralabials, and dorsal

tubercle shape (not trihedral vs. enlarged trihedral). It is

different from H. robustus by larger body size in both

sexes combined (48-63 vs. 32—50) and in males (54.7 vs.

41.8) and females (56.4 vs. 43.6); more lamellae under

Amphib. Reptile Conserv.

rie | as

Pras [3s

029 |

ee ee M

the 1% (8.5 vs. 6.1) and 4" (12.8 vs. 10.1) digits of the

pes; more supralabials (11.8 vs. 9.4); and greater number

of precloacal pores (7.4 vs. 6.1) [Carranza and Arnold

2012: Smid et al. 2013a, 2015] (Table 2). Different from

H. achaemenidicus sp.n. by larger body size (48-63 vs.

28-39), tail, dorsal tubercle rows, number of lamellae

under digits of pes, labials, and postmentals (see Table

2). Different from H. flaviviridis by presence of dorsal

tubercles and without femoral pores. More comparisons

are shown in Table 2

In this section H. sassanidianus sp.n. is_ briefly

compared with other Hemidactylus species outside of

Iran. Different from H. dawudazragi and H. shihraensis

by body size (48-63 vs. 40-49 and less than 49,

respectively). Different from H. asirensis by larger body

size (48.3-63.3 mm vs. 43-48.5 in males, 54.5—57.4

mm vs. 38.3—51.1 in females) and HL/SVL (28-31%

vs. 23-28%). Different from H. alfarraji by precloacal

pores (6-8 vs. 4) [Smid et al. 2016]. Different from H.

kurdicus by postmentals (24 vs. 1) [Safaei-Mahroo et

al. 2017]. Different from H. lavadeserticus by enlarged

keeled tubercles on back (vs. not so enlarged). Different

from H. foudaii by precloacal pores (6-8 vs. 9) and well

developed dorsal and tail tubercles (vs. less developed

and protuberant dorsal, and particularly, tail tubercles).

Different from H. homoeolepis, H. masirahensis, and H.

paucituberculatus by having keeled tubercles on dorsum

(vs. without tubercles on dorsum). Different from H.

inexpectatus, H. endophis, H. hajarensis, H. yerburii,

H. shugraensis, H. yerburii yerburii, H. montanus, H.

awashensis H. minutus, H. homoeolepis, H. mindiae,

H. lemurinus, and H. granosus by number of precloacal

pores (6-8 vs. 4, 14, 4-6, 12.8, 5, 13.7, 11.2, 4.5, 5.8, 4.3,

4, 6, 5.6, respectively) [Smid et al. 2013a, 2015, 2016;

Vasconcelos and Carranza 2014; Carranza and Arnold

2012]. Different from H. /uqueorum and H. homoeolepis

by body size (55.4 vs. 76.8, 31.8) [Carranza and Arnold

2012]. Different from H. turcicus by postmentals (2-4

vs. 2), more longitudinal tubercles on dorsum (15.5 vs.

December 2019 | Volume 13 | Number 2 | e209

c [ek er i

pbs aay aye ts Per id aae fest Raper * > i

ee a aS Le &

Fig. 10. Dorsal and ventral view of (a, b) holoty

13.8), more lamellae under the 1% (8.5 vs. 6.5) and 4"

(12.8 vs. 9.7) digits of the pes, and more supralabials (11.8

vs. 8.2) and infralabials (8.6 vs. 6.7) [Smid et al. 2013a].

Different from H. sinaitus by larger body size in males

(54.7 vs. 39.5) and females (56.4 vs. 45.6), more lamellae

under the 1% (8.5 vs. 5.7) and 4" (12.8 vs. 9.7) digits of

the pes, and more supralabials (11.8 vs. 8.7) [Carranza

and Arnold 2012]. Different from H. jumailiae by more

supralabials (11.8 vs. 9.8), more lamellae under the 1 (8.5

vs. 6.9) and 4" (12.8 vs. 10.9) digits of the pes (Smid et

al. 2013a). Different from H. festivus, H. alkiyumii, and

H. saba by more longitudinal tubercles on dorsum (15.5

vs. 13.3, 12.9, 14) [Smid et al. 2013a; Carranza and

Armold 2012]. Different from H. ulii, H. mandebensis,

and H. adensis by larger body size in males (54.7 vs. 38.6,

41.5, 34) and females (56.4 vs. 40.1, 35, 36.7), and more

longitudinal tubercles on dorsum (15.5 vs. 14.1, 13.3,

aes

Amphib. Reptile Conserv.

ZFMK 97757

aes fies ss

i oa,

: i ere ee a

pe and (c,d) paratype sp cimens of H. pseudoromeshkanicus sp.n.

14) [Smid et al. 2013]. Different from H. /emurinus, H.

masirahensis, H. inexpectatus, H. paucituberculatus,

H. homoeolepis, H. leschenaultii, and H. flaviviridis by

having numerous enlarged tubercles on upper surface of

body (vs. no enlarged tubercles on upper surface of body).

Hemidactylus pseudoromeshkanicus sp.n. (Figs. 10—

12)

urn: Isid:zoobank.org:act: ACECB18C-9C39-4270-A404-BCD88DFCAA52

Holotype

ZFMK 98578, adult male, collected on the western

slope of central Zagros Mountains, Kole-Saat region

Andimeshk, Khuzestan Province, western Iran on 14

June 2010 (32°52’N, 48°43’E, altitude 607 m asl) by

Farhang Torki.

December 2019 | Volume 13 | Number 2 | e209

Three new Hemidactylus species from Iran

Fig. 11. Comparison of postmentals (PM). (a) Three well developed PM in H. romeshkanicus (Holotype, ZMB 75020) and (b) two

postmentals of H. pseudoromeshkanicus sp.n. (Paratype, ZFMK 97757).

Paratype

ZFMK 97757, adult female, same data as for holotype.

Diagnosis

A medium sized Hemidactylus, snout-vent length at

least 74 mm; tubercles distributed all over the dorsum

(except for arms); back with enlarged trihedral keeled

tubercles; granules (rather than scales) cover head and

extend to neck, and rarely to forelimbs; without femoral

pores; precloacal pores present; tubercular heterogeneity

present on dorsum (proximal and distal parts), limbs,

neck, head, and dorsolateral; six tubercles in all whorls

of tail; two postmentals; more lamellae under fingers;

subcaudal scale enlarged; ventral scales not imbricate,

and the ends of ventral scales are simple (cycloid at mid-

part; weakly denticulate at distal and proximal parts of

ventral); enlarged scansors beneath fingers, scansors are

mostly divided, terminal scansor is single; intermixed

color pattern on dorsal body; sexual dichromatism (in

both dorsal and ventral body) occurs between male

(holotype) and female (paratype).

Description of Holotype (Fig. 10a—b)

Measurements (in mm): body size: 75.2; tail length:

88.7; interlimbs: 30.8; head width: 15.2; head length:

23.4: head depth: 8.7; eye-eye: 8.8; ear opening: 3.2; eye

diameter: 5.4; forelimbs length: 29.9; hind limbs length:

33:3.

Body depressed; body, as well as head flattened:

tail more or less flattened; head triangular-shaped; two

postmentals, the first postmentals are enlarged and are

in contact, the 2" postmentals behind the 1‘ enlarged

postmentals; the 1 postmentals are in contact with the

1*t infralabials, the 2"¢ postmentals are in contact with

the 1* and the 2" infralabials, nine scales between 2™

postmentals; infralabials: nine; supralabials: 11; nostril

surrounded by five scales (the 1* infralabial, rostral,

three postnasals); nasals not in contact and separated

by one small scale; ear openings are falcate-shaped,

Amphib. Reptile Conserv.

and horizontal; 19 scales between nostril and eye; 20

scales between eye and ear; rostrum covered by large

granules; between eyes covered by small granules, and

small tubercles (simple and rarely pointed) distributed

among them; upper head covered by smallest granules

and small pointed tubercles distributed among them;

tubercles above ears pointed; tubercles on occipital

mostly pointed and less keeled (heterogeneous);

tubercles on neck pointed and keeled (heterogeneous);

from rostrum to neck covered by granules; tubercles

distributed on dorsum, head, and limbs; tubercles not

found on arms; most body tubercles are keeled; dorsal

tubercles are enlarged, mostly trihedral and strongly

keeled, some of them pointed especially between limbs

(cross view); keeled tubercles between hindlimbs (cross

view: proximal dorsum) intermixed with small and

moderate pointed tubercles, tubercles heterogeneous

(small, large, keeled, pointed, simple) obvious on distal

dorsum (near tail); tubercles on femur mostly trihedral

and keeled (mostly scale-like, different from tubercles on

dorsum); tubercles on forearm are keeled (scale-shape,

different shape from tubercles on dorsum), pointed

and simple (heterogeneous in size and shape); size of

the forearm tubercles smaller than hindlimb tubercles;

scales on palm and sole are granule-like; 16 regular rows

of tubercles on back; 11—13 small tubercles (simple or

pointed) between interorbits; 23 enlarged interlimb

tubercles; 16-17 scales surround each dorsal tubercle;

4—S scales between dorsal tubercles; tail is original; 53

enlarged imbricate scales on subcaudal; last part of tail

cycloid-shape and covered by raised scales; 22 crossbars

on dorsum of tail, 1-3 crossbars are irregular and other

crossbars are regular; tubercle whorls only found in first

part of tail, five scales between each whorl, six tubercles

in 1% whorl, six tubercles in the 2", six tubercles in 3",

and six tubercles in the 4" whorl, after the 5“ whorl

tubercles become very small (six in each whorl); ventral

scales are not imbricate and their size at mid-body are

larger than in other regions, the ends of ventral scales are

December 2019 | Volume 13 | Number 2 | e209

Torki

Fig. 12. Type locality of H. pseudoromeshkanicus sp.n. in

Kole-Saat, Andimeshk, Khuzestan province.

simple (mostly cycloid, not denticulate); 12 precloacal

pores; without any femoral pores; enlarged scansors are

plate-like, terminal scansor is unique (not paired); 1‘

scansor in most fingers 1s unique; lamellae on fingers as

follows: 1s: 11 (1-3 undivided), 2": 11, 3°: 12, 4: 13,

5: 13 (1-3 undivided); lamellae on pes as follows: 1*:

11 (1-3 undivided), 24: 12, 3": 13, 4%: 15, 5": 15; claws

in front of scansors; palm and sole covered by granule-

like scales.

Coloration: irregular grayish pattern covers most of

dorsum extending onto dorsolaterals; occipital covered

by one spotted-bar that extends into eyes; snout is light

grayish; neck region covered by one great grayish spotted-

bar; forearm covered by small gray spots; hindlimbs

covered by light irregular bars that are in contact with

one another; proximal tail covered by irregular bars that

are in contact together, black bars cover distal tail; arm

is without spots; dorsal view of hindlimb digits darker

than forelimb digits; chin is yellowish and light red; color

of ventrum more or less yellowish, without any spots or

bars; palms of digits (hindlimbs and forelimbs) are ashy.

Pattern 1s similar to the live specimen and all spots and

bars are obvious in preserved specimens; the preserved

specimen is colorless.

Description of Paratype (Figs. 2c—d, 11b)

Measurements (in mm): body size: 74.2; tail length: not

original; interlimbs: 31.7; head width: 14.6; head length:

22.8: head depth: 9.1; eye-eye: 9.8; ear opening: 3.2; eye

diameter: 5.1; forelimbs length: 27.4; hind limbs length:

32.4.

Most data are similar to holotype, but some small

differences as follows: 11 scales between 2™ postmentals;

between eyes covered by small granules, and small

tubercles (simple, pointed, and rarely keeled) distributed

among them; upper head covered by smallest granules

and small pointed (rarely keeled) tubercles distributed

among them; tubercles on neck are less pointed and

mostly keeled (heterogeneous); 17 regular rows of

tubercles on back; 18 enlarged tubercles between fore-

and hindlimbs; 16-18 scales surround each dorsum

Amphib. Reptile Conserv.

tubercle; tail 1s missing (most part), zigzag form (without

any crossbars), tubercle whorls only found in first part of

tail, six tubercles in all whorls, 6—7 scales between each

whorl, whorl tubercles distinct by 1-3 scales; ventral

scales are not imbricate and their sizes at mid-body are

larger than in other regions, the ends of ventral scales

are simple (cycloid at mid-part; weakly denticulate at

distal and proximal parts of ventral); without precloacal

pores; without any femoral pores; enlarged scansors are

plate-like, terminal scansor is unique (not paired); first

scansors of most fingers are unique; lamellae on fingers

as follows: 1 and 24: 11, 3": 12, 4 and 5": 13; lamellae

on pes as follows: 15: 11, 2": 13, 3%: 14, 4" and 5%: 15.

Color pattern: intermixed irregular (in contact)

black and grayish pattern covers most parts of dorsum

that extend onto dorsolaterals; bar and inter-bar cover

proximal and distal dorsum; an irregular black stripe

extends to eyes; neck region covered by one great black

bar; one narrow black stripe between eyes and nostrils;

one wide black stripe between eye and ear which extends

to occipital region; forearm covered by small gray spots,

hindlimbs covered by irregular bars that are in contact

with one another; tail covered by irregular bars that are

in contact (without crossbar on tail); arm is without

spots; in dorsal view hindlimb digits strongly darker

than forelimb digits; chin is yellowish, color of ventrum

is light, without any spots or bars; palms of digits

(hindlimbs and forelimbs) are white or less ashy. Pattern

is similar to the live specimen and all spots and bands are

obvious in preserved specimen. The preserved specimen

is colorless.

Habitat and Ecology (Fig. 12)

Specimens belonging to Hemidactylus pseudo-

romeshkanicus sp.n. were collected from the Kol-e-Saat

region, Andimeshk, Khuzestan province. Kol-e-Saat

Region is located between Lorestan-Khuzestan Provinces

and has warm climatic conditions; it is located between

the central Zagros Mountains and Khuzestan Plain. Oak

(Quercus brantii) forest is distributed in the mountains

of this region. The new Hemidactylus specimens show

nocturnal activity, and feed on small insects and insect

larvae occurring in the habitat. Individuals of the new

species actively climb on rocks, and specimens were

collected on rocks during the middle of the night.

Distribution

Presently, this new species is only recorded from

the type locality at Kol-e-Saat region, Andimeshk,

Khuzestan Province, Iran. In spite of several field trips

to areas adjacent to the type locality, no specimens

belonging to this new taxon were found. But based on

geomorphological patterns of the folded mountains of the

western slope of Zagros Mountains, the main distribution

of H. pseudoromeshkanicus sp.n. is expected to extend

towards the mountains of northern Khuzestan province.

December 2019 | Volume 13 | Number 2 | e209

Three new Hemidactylus species from Iran

Sympatric Lizards and Snakes

From the type locality the following additional reptile

species were recorded: Asaccus nasrullahi, Cyrtopodion

scabrum, and Pseudocerastes fieldi.

Etymology

The name “pseudoromeshkanicus” is an allusion to its

similarity to H. romeshkanicus. The color pattern of

this new species appears similar to H. romeshkanicus,

but morphological characters do not match this species,

therefore the prefix “pseudo” is used for the new species.

Comparison with Hemidactylus romeshkanicus

Hemidactylus pseudoromeshkanicus sp.n. 1s significantly

different from H. romeshkanicus by several characters as

follows: two postmentals (instead of three developed in

H. romeshkanicus, Fig. 11); H. pseudoromeshkanicus

sp.n. has more lamellae under 4" digit of pes (13 instead

of nine), 1° digit (11 instead of eight), and 4" digit (15

instead of 12) than H. romeshkanicus (which is slightly

true for other fingers); whorl tubercles on tail (number,

size, and arrangement) as follows: number of tubercles

in each whorl in H. pseudoromeshkanicus sp.n. from 1*

to 4" is unique (6-6-6-6), in contrast in H. romeshkanicus

decreasing number of tubercles from 1 ‘to 4" whorl (7-6-5-

4). scales between each whorl in H. pseudoromeshkanicus

sp.n. more than H. romeshkanicus (5—7 instead of

four); supralabials in H. pseudoromeshkanicus sp.n.

significantly less than H. romeshkanicus (11 instead of

15); tubercle rugosity (in general) on dorsum of body of H.

romeshkanicus is stronger than H. pseudoromeshkanicus

sp.n. (one significant example: three views of trihedral

tubercles show rugosity, that rarely occurs for H.

pseudoromeshkanicus sp.n.), tubercular heterogeneity

(small and large trihedral, pointed) occurs on proximal

and distal part of dorsum of H. pseudoromeshkanicus

sp.n., in contrast to H. romeshkanicus. Nasals separated

by one small scale in H. pseudoromeshkanicus sp.n.,

in contrast, one large scale separates nasals in H.

romeshkanicus.

Comparisons with other Hemidactylus

In general, H. pseudoromeshkanicus sp.n. 1s significantly

different from H. robustus, H. persicus, H. sassanidianus

sp.n., and H. achaemenidicus sp.n. by having mostly

enlarged trihedral tubercles on dorsal body. Differs from

H. robustus in body size (than less 50 vs. at least 74

mm) and tail with more precloacal pores (12 vs. 6—8),

tail tuberculation (keeled and raised instead pointed),

and different dorsal color patterns (irregular bands vs.

spotted). Differs from H. persicus by larger body size

and stronger tubercle rugosity on entire dorsal body and

limbs, head shape and size, and dorsal tubercle rows

(Table 2). Differs from H. flaviviridis by having enlarged

tubercles on dorsum, and without femoral pores. For more

comparisons see Table 2. Differs from H. sassanidianus

sp.n. and H. achaemenidicus sp.n. by having more

Amphib. Reptile Conserv.

precloacal pores (12 vs. 6-8, 6—8, respectively), larger

body size, tail with more dorsal tubercle rows, dorsal

tubercle shape and size (more rugosity and larger in size

for H. pseudoromeshkanicus sp.n.), and more lamellae

under fingers (Table 2).

Brief comparisons show differences of H.

pseudoromeshkanicus sp.n. from other Hemidactylus

spp. outside of Iran. Differs from H. dawudazraqdi,

H. hajarensis, H. homoeolepis, H. jumailiae, H.

Shihraensis, H. alfarraji, H. asirensis, and H. foudaii

by precloacal pores (12 vs. 6-8, 4-6, 3-6, 6-9, 6, 4,

6, 8-10, respectively). Differs from H. kurdicus by

postmentals (2 vs. 1) and precloacal pores (12 vs. 10)

[Safaei-Mahroo et al. 2017]. Differs from H. montanus

by more lamellae beneath 4" digit of pes (15 vs. 9-12).

Differs from H. endophis by large body size (74-75

vs. 59), strongly keeled dorsal tubercles (vs. relatively

weakly keeled), and without femoral pores (vs. 14

pores). Differs from H. /emurinus by presence of well-

developed dorsal tubercles (vs. none). Differs from

H. luqueorum, H. festivus, H. paucituberculatus, H.

lavadeserticus, H. masirahensis, and H. inexpectatus

by more precloacal pores (12 vs. 5-6, 6, 6, 6, 4, and 4,

respectively). Differs from H. turcicus by larger body

size and tail, more lamellae beneath 4" digit of pes (13

vs. 8-11), more precloacal pores (12 vs. 6—10), stronger

tubercular rugosity, and different body color patterns.

Differs from H. mindiae, H. granosus, H. mandebensis,

H. awashensis, H. adensis, H. minutus, H. ulii, H. saba,

H. jumailiae, and H. yerburii, by having larger body

size. Differs from H. alkiyumii by having more rows of

tubercles (16-17 vs. 11-14), more lamellae under the 4"

digit of pes (15 vs. 10-12), and more precloacal pores

(12 vs. 6-10). Body size in H. pseudoromeshkanicus

sp.n. is smaller than in H. aaronbaueri, dorsal tubercles

in H. pseudoromeshkanicus sp.n. are much larger than in

H. aaronbaueri, also, color pattern is different from H.

aaronbaueri. By having enlarged, trihedral, and regular

dorsal tubercles H. pseudoromeshkanicus sp.n. is easily

distinguished from several species of Hemidactylus

including: H. aaronbaueri, H. bowringii, H. brookii, H.

flaviviridis, H. garnotii, H. karenorum, H. leschenaultii,

H. maculatus, H. persicus, H. prashad, H. subtriedrus,

and H. triedrus. Digits are relatively slender in H.

scabriceps, but in H. pseudoromeshkanicus sp.n. they

are broadly dilated. H. sinaitus (from Sudan to Northern

Somalia, and Arabia) has smaller and more widely

separated dorsal tubercles, but H. pseudoromeshkanicus

sp.n. has mostly trihedral tubercles.

Note on Hemidactylus Inhabitants from Iran

Hemidactylus inhabitants of the Iranian plateau have a

complicated history. Anderson (1999) reported three

Hemidactylus (H. flaviviridis, H. persicus, and H.

turcicus) from Iran. Anderson (1974) had recorded H.

garnotii in the fauna of Iran, but in 1999 he excluded

254 December 2019 | Volume 13 | Number 2 | e209

Torki

it from Iran due to incomplete data from I. Darevsky;

and he then diagnosed this species as H. flaviviridis

(Anderson 1999). Anderson collected some Hemidactylus

sp. specimens from southwest Iran that do not to match

H. flaviviridis, H. persicus, or H. turcicus (Anderson

1999). Anderson was concerned that H. brookii might

be distributed in southern Iran, but this species has

not been collected inside Iran. Therefore, based on

Anderson’s studies (1999), four species occur in Iran: H.

flaviviridis, H. persicus, H. turcicus, and Hemidactylus

sp. A molecular study (Bauer et al. 2006a) confirmed the

distribution of H. robustus in southwestern Iran; and,

little difference exists between H. robustus from Iran

on the one hand and from the United Arab Emirates and

Egypt on the other. Firouz (2000) has cited H. flaviviridis,

H. persicus, and H. turcicus for the fauna of Iran. Torki

et al. (2011) showed five Hemidactylus species to occur

in Iran, viz: H. flaviviridis, H. persicus, H. turcicus, H.

robustus, and H. romeshkanicus. Due to this author’s

revision of the gecko fauna of Iran (2016-2020 FTE

program), one previous occurrence of Hemidactylus was

identified as H. turcicus (FTHM005100-5110 in Torki et

al. 2011); however, new morphological evidence shows

that it is completely different from H. turcicus as well as

from H. robustus. As described here, this population (H.

achaemenidicus sp.n.) shows differences in important

taxonomic characters from other Hemidactylus species

both inside and outside of Iran (as well as the arid clade).

Hosseinzadeh et al. (2014) worked on the morphology

of Hemidactylus species of Iran, and their work showed

four Hemidactylus species from Iran: H. flaviviridis,

H. persicus, H. robustus, and H. romeshkanicus, as

they rejected H. turcicus from the Iranian gecko fauna.

Based on recent phylogenetic studies on Hemidactylus,

particularly H. turcicus and H. robustus (Carranza and

Arnold 2012; Smid et al. 2013b, 2015), I suggest that

the H. robustus specimens which were examined by

Hosseinzadeh et al. (2014) do match with both H.

turcicus and H. robustus. They do not show the important

taxonomical characters that are important for diagnosis

of H. turcicus and H. robustus from several of those

populations.

Based on recent molecular studies (Carranza and

Arnold 2012; Smid et al. 2013b, 2015), H. persicus

from Iran shows characteristics of being a separate clade

from Arabian Hemidactylus. This clade shows three

distinguishable species, and one of them (FTHM005110)

is the new Hemidactylus achaemenidicus sp.n. described

here. The locality of FTHM005110 cited in_ that

phylogenetic study is incorrect and must be changed to the

type locality of H. achaemenidicus sp.n. given here. On

the other hand, three specimens of H. persicus (JS103—

5) among the Iranian persicus clade (Smid et al. 2013)

showed more differences from other H. persicus, but the

localities of these specimens were not cited in that paper,

and are nearest to the type locality of H. sassanidianus

sp.n. (see Fig. 4 in Smid et al. 2013). On the other hand,

Amphib. Reptile Conserv.

H. robustus from the coastal Persian Gulf (Bandar-e-

Lenge) is a match with the Arabian H. robustus clade

(Smid et al. 2013b, 2015). The oldest reported dispersal

from Arabia occurred 13.1 Ma, when the ancestor of

H. persicus colonized Iran (Smid et al. 2013b). This

time-frame (13.1 Ma) is perfect for speciation among

the Hemidactylus inhabiting the Iranian plateau as well

as the Zagros Fold-Thrust Belt. A few collections from

the southern part of Iran (mostly coastal Persian Gulf)

show three clades in the phylogenetic tree of Smid et al.

(2013b). Based on the distribution of Hemidactylus inside

the Iranian plateau, here I suggest that Hemidactylus has

several monophyletic clades as well as more species

which remain unknown.

Although some works exclude H. turcicus (e.g.,

Hosseinzadeh et al. 2014, Smid et al. 2014) from the

fauna of Iran, Smid et al. (2014) did not explicitly

reject H. turcicus from Iran (see Map 46), and Smid et

al. concluded that H. turcicus 1s not distributed in Iran.

I disagree with those assessments, and do not exclude

this widespread species from the fauna of Iran until more

comprehensive data about the Hemidactylus inhabiting

Iran (especially from phylogenetic studies) are available.

One important reason supporting the acceptability of H.

turcicus for the fauna of the Iranian Plateau is its wide

distribution in adjacent areas to the west (e.g., Turkey)

and east (e.g., Pakistan) of Iran (e.g., Turgay and Atat

1994; Khan 2006).

Bauer et al. (2006a) identified all populations of

small Hemidactylus as a H. robustus. Some authors (e.g.,

Gholamifard and Rastegar-Pouyani 2011; Hosseinzadeh

et al. 2014) followed that assessment. Based on

phylogenetic analysis, H. achaemenidicus sp.n. 1s

completely distinguishable from H. robustus (e.g., Smid

et al. 2013, 2015). Therefore, there are at least three

distinct species of small Hemidactylus in Iran including:

H. robustus, H. turcicus, and H. achaemenidicus sp.n.

Based on all the studies cited above, all Hemidactylus

species of Iran (except H. flaviviridis) show much

complexity and I classify them here in three groups as

follows: H. persicus-complex (including H. persicus, H.

sassanidianus sp.n., and H. achaemenidicus sp.n.); H.

robustus-complex; and H. romeshkanicus-complex (H.

romeshkanicus and H. pseudoromeshkanicus sp.n.).

In summary, at least eight species of Hemidactylus

are distributed on the Iranian Plateau: H. flaviviridis, H.

persicus, H. robustus, H. turcicus, H. romeshkanicus, H.

sassanidianus sp.n., H. achaemenidicus sp.n., and H.

pseudoromeshkanicus sp.n.

Note on Hemidactylus parkeri Loveridge 1936

H. parkeri was described by Loveridge (1936), but this

Species was downgraded or rejected from subsequent

species lists of Hemidactylus (e.g., Arnold 1980; Smid

et al. 2015) and replaced by H. turcicus and H. robustus.

Based on the following reasons, I do not agree with

December 2019 | Volume 13 | Number 2 | e209

Three new Hemidactylus species from Iran

this decision. (i) Type locality: The type locality of H.

parkeri is very far from the type localities of H. turcicus

(Asiatic Turkey, by Moravec et al. 2011) and H. robustus

(“Egypten, Arabien, und Abyssinien” restricted to “the

Red Sea coast of the State of Eritrea” by Smid et al. 2015).

(ii) Ecology and climate: Loveridge (1936) described his

new species in Zanzibar Island (Tanzania), and this island

may have an important role in the speciation of these

geckos. Additionally, Zanzibar Island is located near the

equator, with special ecological and climatic conditions;

and the ecological and climatic conditions of the type

locality of H. parkeri are completely different from the

type localities of H. turcicus and H. robustus. (111) New

methods and insights: Based on phylogenetic studies,

most Hemidactylus species described long ago have been

split into several species, such as H. persicus, H. yerburii,

H. turcicus, and H. robustus (e.g., Carranza and Arnold

2012; Smid et al. 2013, 2015). Therefore, additional

phylogenetic studies on the equatorial Hemidactylus

species are necessary to resolve this problem. (iv) Six

species of Hemidactylus are distributed in Tanzania, and

H. parkeri is not synonymous with all of them (Uetz

2019). On the other hand, only one species is endemic to

Tanzania (H. tanganicus). Based on the above reasons,

there is not a logical and scientific basis for the rejection

of H. parkeri. Therefore, in this study I am in agreement

with Lazza (1978, 1983) on the validity of H. parkeri.

Note on Gecko Conservation in Iran

Based on observations during 20 years, two main threats

for the geckos of Iran are apparent: (1) Rumor: People

in this region believe that geckos are poisonous and fear

them, especially in cities and less so in villages. This

rumor applies to all geckos inhabiting human homes. (11)

Trade: Among geckos, the fat-tailed gecko (Eublepharis)

is an important species that is sold. Eublepharis 1s

considered attractive and some people find it interesting

as a pet. During recent years, trade of this gecko has

increased among the Iranian people. Although 47% of

geckos inhabiting Iran belong to the Red List, the IUCN

category (http://www.iucn.org/) of most is LC (or Least

Concern). The geckos in Iran have the best conservation

situation compared to other amphibians and reptiles, and

their nocturnal activity may have an important role.

Key to Hemidactylus Species Distributed in Iran

la: Dorsal tubercles absent, femoral pores present

LEN ore ne en PL ee ve ee H. flaviviridis

1b: Dorsal tubercles present, femoral pores absent....2

2b: 9-11 precloacal pores...........................H. persicus

We Mls preclOacals POLES cat een tate cick Ca Raat 5

3a: 2-4 postmentals, not small Hemidactylus (body

Amphib. Reptile Conserv.

size is more than 48 mm)...... H. sassanidianus sp.n.

3b: Two postmentals, small Hemidactylus (less than 50

4a: Females have less rugosity than males, subcaudals

covered by small scales and/or plate-like scales (few

in number: 0—22)............... H. achaemenidicus sp.n.

4b: Sexual rugosity does not occur (females have

approximately full rugosity), subcaudal scales enlarged

(nopsimalll-seales),, .) 2s. 0/ boda dege o® H. robustus

5a: Two postmentals.....H. pseudoromeshkanicus sp.n.

Sbe Three; postmentals:....of Ree: H.. romeshkanicus

Acknowledgments.—1 wish to thank Professor Steven

Anderson (California, USA) for editing and improving

my manuscript, and Craig Hassapakis (Utah, USA) and

Michael L. Grieneisen (California, USA) for improving

my manuscript. I wish to thank Professor W. Bohme

and his collaborators in ZFMK (ZFMK, Germany) for

marked type specimens.

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Amphib. Reptile Conserv.

Farhang Torki earned his B.Sc. degree in Animal Biology from Lorestan University,

Iran, and his M.Sc. degree in Animal Biosystematics from Razi University in Iran.

During his B.Sc. studies, Farhang worked on histological and embryological methods,

particularly as applied to the spermatogenesis and oogenesis of reptiles, and the herpe-

tofauna of Lorestan Province. During his M.Sc. studies he worked on the systematics

of amphibians and reptiles of the southern and western Iranian Plateau, and continued

his developmental biology work in herpetology. Following his graduate work, Farhang

established (2006) the Farhang Torki Ecology and Herpetology Center for Research

(FTEHCR), the Farhang Torki Herpetology Museum (FTHM), and recently Biomatical

Center for Researches (BMCR) from 2018. Currently, Farhang is studying evolution

and developmental biology, based on mathematical methods. Iran University of Medi-

cal Sciences supported his research during 2018.

258 December 2019 | Volume 13 | Number 2 | e209

Official journal website:

amphibian-reptile-conservation.org

Amphibian & Reptile Conservation

13(2) [General Section]: 259-264 (e210).

The Blue Dyeing Poison-Dart Frog, Dendrobates tinctorius

(Dendrobates azureus, Hoogmoed 1969): extant in Suriname

based on a rapid survey

'*Christian A. d’Orgeix, 'De’Jah Hardy, ‘Sarah Melissa Witiak, 7Laren R. Robinson, and *Rawien Jairam

'Department of Biology, Virginia State University, Petersburg, Virginia, 23806, USA *Department of Agriculture and Ecology, Virginia State

University, Petersburg, Virginia 23806, USA *National Zoological Collection of Suriname, Anton de Kom Universiteit, Paramaribo, SURINAME

Abstract.—The blue color morph of Dendrobates tinctorius, originally described as D. azureus, has only been

reported from a few forest islands surrounded by the Sipaliwini savanna in Suriname, South America. The last

published survey of these populations occurred in 1996. The threats of emergent diseases, illegal collecting,

climate change, and habitat destruction through anthropogenic fires toward these populations are unknown.

This report presents the results of a rapid survey of the three forest islands where D. tinctorius historically

occurred to assess its current status. One 50 x 50 m survey plot was established in each of these three forest

islands. A total of 23 frogs were recorded, with some individuals found in each of the forest island surveyed.

These results indicate that D. tinctorius populations are still present at all three historical localities surveyed

by previous researchers in 1968 and again in 1996, although the current surveys found fewer frogs. During the

surveys there was no evidence of illegal collecting or habitat degradation. These observations provide baseline

data that can be used for future monitoring and protection of one of the most geographically restricted and

unique color morphs of D. tinctorius.

Keywords. Amphibia, Dendrobatidae, conservation, Sipaliwini savanna, population decline, habitat fragmentation,

climate change

Citation: d’Orgeix CA, Hardy D, Witiak SM, Robinson LR, Jairam R. 2019. The Blue Dyeing Poison-Dart Frog, Dendrobates tinctorius (Dendrobates

azureus, Hoogmoed 1969): extant in Suriname based on a rapid survey. Amphibian & Reptile Conservation 13(2) [General Section]: 259-264 (e210).

4.0 International (CC BY 4.0): https://creativecommons.org/licenses/by/4.0/], which permits unrestricted use, distribution, and reproduction in any

medium, provided the original author and source are credited. The official and authorized publication credit sources, which will be duly enforced, are

as follows: official journal title Amphibian & Reptile Conservation; official journal website: amphibian-reptile-conservation.org.

Received: 10 May 2018; Accepted: 21 February 2019; Published: 18 December 2019

Introduction

The Blue Dyeing Poison-dart Frog was described a

half-century ago as Dendrobates azureus (Hoogmoed,

1969) and subsequently synonymized with D. tinctorius

(Wollenberg et al. 2006). The inclusion of this species

as a color morph of D. tinctorius was also confirmed by

Noonan and Gaucher (2006). Ouboter and Jairam (2012)

considered this species to be a subspecies of D. tincto-

rius due to the geographically isolated occurrence of this

particular morph. Its classification as a subspecies was

refuted by Frost (2017) who maintained this species as

D. tinctorius. Other blue morphs of D. tinctorius were

documented by Silverstone (1975) from Shudikar-wau,

Guyana and by Avila-Pires et al. (2010) from Esta¢ao

Ecolégica Grao-Para North, Brazil. However, specimens

from Suriname do not display the typical light D. tincto-

rius pattern on the dorsum, whereas specimens from the

other mentioned locations still have this pattern.

The predominantly blue color morph (“azureus”) found

in Suriname is restricted to a few isolated “forest islands”

Correspondence. *cdorgeix@vsu.edu

Amphib. Reptile Conserv.

in the Sipaliwini savanna in southern Suriname (Hoog-

moed 1969; Riezebos 1979; Cover 1997; Ouboter and Jai-

ram 2012). This area was mapped in July 1935 when an

expedition by Baron van Lynden was charged with estab-

lishing the southern border of Suriname (Lynden 1939).

The Sipaliwini savanna ts part of a larger savanna complex

covering portions of Suriname and northern Para, Brazil

(Paru savanna). The current vegetation primarily consists

of grasses, sedges, scattered shrubs, trees, and Mauritia

palms (Oldenburger et al. 1973). The sharp demarcation

between the savanna and forest islands may be due to geo-

logical factors as suggested by Hoogmoed (1973), while

the anthropogenic influence of fires set during the dry sea-

son by the indigenous people might have been a reason

that the savanna remained open (Oldenburger et al. 1973;

Ouboter and Jairam 2012) [Fig. 1].

A number of factors potentially threaten the continued

existence of these populations of D. tinctorious in the wild.

Their extremely restricted range and small population

size are factors that increase the risk of extinction (Hall

et al. 2008). Anthropogenic fires could potentially reduce

December 2019 | Volume 13 | Number 2 | e210

The Blue Dyeing Poison-Dart Frog extant in Suriname based on a rapid survey

Fig. 1. Map of Suriname, South America (upper left inset) with approximate location of Sipaliwini savannah indicated by the black

box. Forest Islands are outlined in red and identified by numbers corresponding to those of Hoogmoed (1969, 2019). Arrows indicate

locations of the 50 x 50 m plots surveyed in Forest Islands 1, 2, and 4 during 16—18 June 2015. Forest Island 3 was not surveyed. In

this 2004 Google image, savanna vegetation surrounding the forest islands had been recently burned by indigenous hunters.

the size of the forest islands, and charred trees on the

periphery of the forest islands were noted by Cover (1997).

The emerging infectious disease Batrachochytrium

dendrobatidis (Bd) may put these populations at risk

(Courtois et al. 2012, 2015; James et al. 2015). Although

Bd has not been documented in Suriname, it has been

documented in D. tinctorius in French Guiana (Courtois et

al. 2015) which is east of Suriname. In addition, the blue

color morph of D. tinctorius occurring in the Sipaliwini

savanna is potentially vulnerable to rarity-fueled

exploitation for the pet trade (Hall et al. 2008).

Despite having been discovered almost 50 years ago

(Hoogmoed 1969), the only reported subsequent attempt

to ascertain the status of the blue morph of D. tinctorius in

the Sipaliwini savanna occurred in 1996 (Cover 1997). At

that time Cover (1997) found frogs but reported potential

evidence of illegal collection. Thus, the primary objective

of the current study was to determine the current status

(presence or absence) of D. tinctorius, in the Sipaliwini

savanna. If present, the plan was to establish survey plots

and conduct preliminary surveys to serve as benchmarks

for future population studies.

Materials and Methods

Survey plots. A survey plot measuring 50 x 50 m was

established in each of three forest islands which were

Amphib. Reptile Conserv.

separated by approximately 350-750 m of tall grass

savanna. Forest islands were identified using the same

numbering scheme as Hoogmoed (pers. comm.) [Fig.

1]. Survey plots within the forest islands measured

approximately 4% of 6 ha (Forest Island 1), 12.5% of

2 ha (Forest Island 2), and 6% of 4 ha (Forest Island

4) [Fig. 1]. Forest Island 3, which lies ~300 m north of

Forest Island 2, was not surveyed due to time constraints.

Plot placements were determined by entering each of the

forest islands and visually searching until a D. tinctorius

was encountered (Fig. 2). The nearest stream was then

used as one side of the 50 x 50 m sample plot, based

on previous research by Hoogmoed (1969) and Cover

(1997) citing stream preference by the frogs. Plot corners

were marked with GPS coordinates using a Garmin

60CSx. All three forest islands were subsequently

digitized using images from Google Earth Pro, which

were further edited in ArcMap, version 10.2, to show

the locations where the surveys were conducted. To do

this, Google Earth imagery was acquired and added as

a layer into ArcMap. Then Arc toolbox's KML layer

feature was used to transfer the digitized forest islands

created in Google Earth to ArcMap. The resulting image

emphasizes the presumed isolation by habitat of the D.

tinctorius populations from one another (Fig. 1).

Survey plots 1 and 4 were bordered on three sides by

small clear water streams. Survey plot 2 was characterized

December 2019 | Volume 13 | Number 2 | e210

d’Orgeix et al.

- 2: F ; :

Fig. 2. Dendrobates tinctorius from Sipaliwini savanna,

Suriname.

by having a small clear water stream at the lowest part

of the plot. Survey plot 1 was located on a north-facing

Slope that rose from 315-340 m in elevation. The area

was under a fully closed canopy forest with dense

understory vegetation interspersed with lianas. The forest

floor was covered by leaf litter, decomposing fallen

trees, and boulders of about 2—3 m in height and 3-6 m

in diameter (Fig. 3). Survey plot 2 was on a southeast

facing slope that rose from 315-325 m in elevation. The

plot was under a closed canopy with an open understory

and a forest floor covered by leaf litter. There were no

boulders in this plot. Survey plot 4 was on a west-facing

slope that rose from 300-350 m in elevation. The area

was under a fully closed canopy with dense understory

vegetation. One-third of the plot on the southwest side

had a dense understory of Phenacospermum sp. and

Poaceae bamboo species. The forest floor was covered

by leaf litter interspersed with 10-15 clumped boulders

that were about 1—3 m high and 1-5 m in diameter.

Survey methodology. Each plot was visually surveyed

once, for varying amounts of time, from 1030-1300 h.

Surveys occurred on 16—18 June 2015, three days after

the last rainfall on 13 June 2015. To survey a plot, nine

individuals were spaced approximately 1.5 m alongside

each other forming a line. Starting at one corner of the

plot, the surveyors walked 50 m to the opposite side of

the plot. Upon reaching the far side the surveyors again

spaced laterally starting 1.5 m from the person on the

farthest end to cover the next section, and walked back

towards the opposite side. This process was repeated until

the whole plot was surveyed. Survey participants wore

sterile disposable gloves which were changed between

the captures of each individual. Capture locations of

individual frogs were recorded as GPS coordinates using

a Garmin 60CSx. Frogs were then placed in individually

labeled Zip-Lock bags. Snout-vent length (SVL) was

measured to nearest 0.1 mm with digital calipers. Adults

were not sexed. After measurements were taken, each

frog was released at its original point of capture.

To identify potential changes in perimeter sizes of

the forest islands due to environmental or anthropogenic

Amphib. Reptile Conserv.

Fig. 3. Stream and boulder habitat in Survey Plot 1, Sipaliwini

savanna, Suriname.

factors, the most historical and most recent available

Google Earth images from 31 December 1969 and 17

November 2004 were used. These images were visually

compared by juxtaposing the forest islands outlined in

red, from 2004 (Fig. 4A) onto the 1969 image (Fig. 4B).

Statistical methods. A General linear model ANOVA

was used to compare SVL values between specimens

from each forest island. A f-test was used to compare

SVL of frogs measured in the current survey with those

reported by Hoogmoed (1969: Table I). SVL are reported

to nearest 0.1 mm + 1 SD. All analyses were conducted

using Minitab 17 statistical software (Minitab Inc., State

College, Pennsylvania, USA).

Results

Twenty-two adults and one juvenile were found in the

three forest island plots surveyed. Sixteen frogs were

found during the surveys, and seven were found adjacent

to the survey plots. Two frogs were found in Forest

Island 1 in 1 h (270 person-min/frog). Four frogs were

found adjacent to the survey plot in Forest Island 1 in

0.5 h (67.5 person-min/frog). Four frogs were found in

Forest Island 2 in 0.5 h (67.5 person-min/frog). Ten frogs

were found during the survey of Forest Island 4 in 1.5 h

(81 person-min/frog). Three frogs were found adjacent to

the survey plot in Forest Island 4 in 0.5 h (90 person-min/

frog). The overall mean yield of encounters was 115.2

person-min/frog. Variations in time spent conducting a

survey often reflected the difficulty in traversing the plot

based on the physical obstructions encountered (e.g.,

boulders and density of herbaceous undergrowth).

Adult SVL values ranged from 36.7-46.4 mm, with

a mean of 42.0 + 3.1 mm. No significant differences

between SVL of adult frogs from different forest islands

were found (one-way ANOVA: F,,,, = 0.23, P = 0.795).

No significant difference was found in a comparison

of SVL values recorded by Hoogmoed (1969) and the

current survey (t-test: t= 1.49, df= 44, P = 0.144). The

single juvenile recorded here measured 18.9 mm SVL.

A comparison of the vegetative perimeters of Forest

December 2019 | Volume 13 | Number 2 | e210

The Blue Dyeing Poison-Dart Frog extant in Suriname based on a rapid survey

ae rs

oa oie ae

\ he OE ee , 2 ae

eo te, a

Fig. 4. Forest Islands 1, 2, and 4, extant vegetation com

ze = kh

parison between Google Earth images for 2004 (A) and 1969 (B). Red

outlines delineating the three forest islands were juxtaposed from the 2004 image onto the 1969 image.

Islands 1, 2, and 4 appear almost identical in the two

Google Earth images taken in 2004 (Fig. 4A) and 1969

(Fig. 4B).

Discussion

Survey findings. The survey sites represented three of the

four forest islands where frogs were found in the surveys

by Hoogmoed (1969) and Cover (1997). In the current

surveys only adults were found, with the exception of

a single juvenile. No males transporting tadpoles were

found. Hoogmoed (1969) reported a male carrying two

tadpoles on his back on 30 September 1968, while Cover

(1997) who surveyed in June, did not note this behavior,

although he reported newly metamorphosed individuals

and tadpoles at two sites.

This survey recorded fewer frogs (23) than either

Hoogmoed (1969) or Cover (1997). Cover (1997, pers.

comm.) reported 56 frogs from Forest Island sites 1, 2,

and 4. Hoogmoed (1969, 2019) reported 82 frogs from

Forest Islands 1-4, during 11 different dates in 1968 and

1970. Differences in methodologies, season, and weather

limit most direct comparisons between these studies.

Both Cover (pers. comm.) and Hoogmoed (1969, 2019)

concentrated their searches in suitable habitat along

stream courses, while in the current survey the 50 x

50 m plots were centered around the spot where a frog

was first encountered. Weather events, such as rainfall,

presumably influence frog activity. For example, Cover

noted that after heavy rains at one site, the numbers of

frogs visible on the forest floor increased dramatically

(J. Cover, pers. comm.). A similar situation was also

reported by Wevers (2007). The current surveys took

place 3-5 days after the last rains. Hoogmoed (1969,

2019) reported 82 frogs during herpetological inventories

conducted on 11 different days in September and October

1968 and February 1970. In comparing the time required

to encounter a frog, Hoogmoed (2019) averaged 10.1

person-min/frog while the average in the current survey

was 115.2 person-min/frog. This would suggest a

Amphib. Reptile Conserv.

significant population decline if all other factors were

equal.

The blue morph of D. tinctorius in the Sipaliwini

savanna forest islands appears to be both geographically

and genetically isolated from other color morphs of D.

tinctorius 1n Suriname. The nearest known populations

of D. tinctorius, which are black and yellow dorsally, are

approximately 23 km north of the closest forest islands

surveyed (Fouquet et al. 2015). Blue morphs of D.

tinctorius in Sipaliwini are more than 300 km away from

the blue morphs reported by Silverstone (1975), and

approximately 315 km from the blue morphs reported by

Avila-Pires et al. (2010). In addition, the frogs in each of

the forest islands surveyed may be genetically isolated

from each other. Exploration of other remote forest

islands in the Sipaliwini savanna region may reveal

additional isolated populations of blue D. tinctorius.

Future molecular analyses could elucidate the genetic

divergence in these forest island populations since their

presumed isolation approximately 10,000 years ago

(Riezebos 1979).

Conservation considerations and future threats.

Batrachochytrium dendrobatidis (Bd) has been recorded

in French Guiana (Courtois et al. 2012, 2015) and Brazil

(Becker et al. 2016) which border Suriname to the east

and south, respectively. During this rapid survey no

dead frogs or frogs displaying visual symptoms of Bd

were found, although seemingly healthy specimens are

still known to be carriers of this disease (Coutinho et al.

2015).

While no evidence was found of illegal collecting for

the pet trade, that lack of evidence is not proof that it does

not occur or will not occur in the future. The indigenous

Trio peoples control both access to the transportation to

the savanna (canoes) and permission to visit the forest

islands where the frogs are found. Trio culture recognizes

the uniqueness of these frogs. Permission from the

village chief must be granted to visit the sites where frogs

may be observed, but collection 1s not allowed. This is

December 2019 | Volume 13 | Number 2 | e210

d’Orgeix et al.

enforced by the accompanying Trio guides. Presently,

the Trio people’s stewardship practices provide a large

measure of protection from collecting for the pet trade.

Sustainable programs that benefit the Trio community

should be implemented to reinforce their stewardship

of the habitat and wild populations of this unique color

morph.

Visual comparison of the perimeter size of these

forest islands between the 2004 and 1969 images do

not appear to indicate a significant change in size,

despite the history of anthropogenic fires (Fig. 4A,B).

Cover (1997) reported charred wood on the periphery

of the forest islands. However, when he observed a

fire in the savanna, it stopped when it reached the lush

vegetation on the perimeter of a forest island (J. Cover,

pers. comm.). Although the forest islands appear to be

stable in size, due to their small area and isolation, any

reduction in the water sources either through drought

or climate change could threaten both the forest island

vegetation and associated stream habitat. An increase in

xeric conditions could increase the risk of forest island

vegetation becoming drier on the periphery and thus more

susceptible to anthropogenic fires resulting in the edges

of these islands moving inwards, leading to a decrease in

their size or even complete destruction.

Barring direct human collection, climate change may

be the most serious threat to this unique blue morph of D.

tinctorius. There 1s a growing body of evidence that tropical

species will need to undergo elevational or latitudinal

range shifts to remain in analogous climatic conditions in

the future (Colwell et al. 2008; Nowakowski et al. 2016).

Given the isolated nature of these forest islands, their lack

of connectivity with either cooler forests or altitudinal

refugia, any change to the forest island habitat may be the

biggest threat to the continued survival of this unique D.

tinctorius blue morph in the wild.

Acknowledgements.—We would like to thank Paul

Ouboter from the National Zoological Collection of

Suriname for support and advice during the time spent

in Suriname. Dallas Davidson, Akira N. Harris, Ahnaia

White, Christian H. d’Orgeix, Sabria Greiner, and Alyssia

Velez participated in surveying the frogs. The Trio

people granted us permission to work in the Sipaliwini

savanna and provided lodging, transportation, and guides

to the sites. Marinus S. Hoogmoed and Jack Cover made

insightful comments to drafts of this manuscript, and

shared unpublished data and information. Tom Mathies

and Paul Kaseloo also provided helpful suggestions.

Permits for the fieldwork were granted by the Nature

Conservation Department of Suriname. This work was

supported in part by a grant to C. d’Orgeix through the

HBCU-UP of the National Science Foundation under

NSF Cooperative Agreement No. HRD-1036286. Any

opinions, findings, and conclusions or recommendations

expressed in this material are those of the author(s) and

do not necessarily reflect those of the National Science

Amphib. Reptile Conserv.

Foundation.

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Christian d’Orgeix is a behavioral ecologist and conservation biologist at Virginia State University.

Christian’s research focuses on the mating systems and conservation biology of reptiles and amphibians.

One of his current projects examines the probability of extinction of high- and low-elevation populations

of lizards. In Suriname, Christian and his students collaborate with Dr. Paul Ouboter and Rawien Jairam

from Anton de Kom Universiteit, Suriname, on studies on the behavior and conservation of frogs and

Rawien Jairam works at the National Zoological Collection of Suriname and is co-author of the book

Amphibians of Suriname. Rawien has an M.Sc. in Conservation Biology and has been interested in the

herpetofauna of Suriname for many years. Apart from general herpetology, he is specifically interested

in taxonomy and species distributions.

De'Jah T. Hardy is a junior Biology major at Virginia State University. De’Jah traveled to Suriname,

South America, to conduct research on frogs and ants. She also traveled to China to take Clinical

Medicine classes at Jinan University in Guangzhou. De’Jah’s goal is to explore as many opportunities as

she can as an undergraduate, and then go straight for her Master’s in Psychology. She plans to travel the

world someday, helping children as a traveling Occupational Therapist.

Sarah Melissa Witiak is a plant biologist who received her Ph.D. from Pennsylvania State University

(University Park, Pennsylvania, USA) and currently teaches at Virginia State University. Sarah is pri-

marily interested in the “pretty” parts of biology, including poison-arrow frogs, insect galls, and flower

evolution and ecology. Her current projects include studies of plant volatiles and a survey of insect gall

Laren Robinson worked as a GIS specialist at Virginia State University. She is currently pursuing

career options in GIS applications. (No photo available)

Amphib. Reptile Conserv.

December 2019 | Volume 13 | Number 2 | e210

Official journal website:

amphibian-reptile-conservation.org

Amphibian & Reptile Conservation

13(2) [General Section]: 265-266 (e211).

Book Review

Night Lizards: Field Memoirs and a

Summary of the Xantusiidae

Howard O. Clark, Jr.

Colibri Ecological Consulting, LLC, 9493 North Fort Washington Road, Suite 108, Fresno, California 93730, USA

Keywords. Behavior, biogeography, ecology, reproduction, reptiles, Scincomorpha, Squamata

Citation: Clark HO Jr. 2019. Book review—Night Lizards: Field Memoirs and a Summary of the Xantusiidae. Amphibian & Reptile Conservation 13(2)

[General Section]: 265-266 (e211).

ternational (CC BY 4.0): https://creativecommons.org/licenses/by/4.0/], which permits unrestricted use, distribution, and reproduction in any medium,

provided the original author and source are credited. The official and authorized publication credit sources, which will be duly enforced, are as follows:

official journal title Amphibian & Reptile Conservation; official journal website: amphibian-reptile-conservation.org.

Received: 11 December 2019; Accepted: 11 December 2019; Published: 12 December 2019

Herpetologist Robert L. Bezy has produced a fascinating

memorr of his life and the lives of night lizards. When

reading about or researching night lizards, Bezy’s name

comes up often. When Bezy began his career there were

only a few night lizards known to science; now there are

35 living species in the family Xantustidae. The book

begins with a brief autobiography, detailing what mo-

tivated Bezy to pursue herpetology and what became a

long and winding road full of exciting discoveries and

working with some of the best herpetologists in the field.

Some of my favorite sections are where Bezy divides his

narrative into “locational highlights” and tells stories of

some experiences that had significant impacts on his life.

These stories are rich in landscape descriptions, wildlife

encounters, and how the adventure—planned or not—in-

fluenced the way he views life. These sorts of reflections

are priceless and allow the reader to reflect on his or her

own life. As I read through these stories I found myself

frequently reminiscing about my various encounters with

night lizards and other wildlife.

Following the autobiography and locational high-

lights, Bezy delves into the historical perspectives re-

garding night lizards. A “must read” is the account of

Janos Xantus and Spencer Fullerton Baird. Baird named

the night lizard genus (Yantusia) and family (Xantusi-

idae) after Xantus and interesting enough Xantus later

wrote to Baird that he didn’t even remember collecting

the small lizard. Other important perspectives include

those of John Van Denburgh, Edward H. Taylor, Hobart

M. Smith, Jay M. Savage, Robert G. Webb, Richard G.

Zweifel, and Charles H. Lowe. All of these men contrib-

uted to the natural history of the night lizards and Bezy

does a splendid job recapping their contributions.

Correspondence. howard.clark.jr@gmail.com

Amphib. Reptile Conserv.

NIGHT LIZARDS

Field Memoirs and a

Summary of the Xantusiidae

Robert L. Bezy

Title: Night Lizards: Field Memoirs and a Summary of the

Xantusiidae

Authors: Tell Hicks (Artist), Robert Bezy (Author)

ISBN-10: 1938850599; ISBN-13: 978-1938850592

Publisher: ECO Herpetological Publishing

Pages: ii + 220; Price: USD $24.95

December 2019 | Volume 13 | Number 2 | e211

Night Lizards: Field Memoirs and a Summary of the Xantusiidae

Fig. 1. Nantusia aes nen the ear Desert, ieee Angeles

County, California. Photo by Howard O. Clark, Jr.

Another highlight of the book is the “questions” sec-

tion. Here, Bezy addresses a variety of natural history

aspects of the night lizard— undoubtedly questions that

he had over the years and now the reader has the oppor-

tunity to read the answers with Bezy as the messenger.

Topics include rock crevice ecology of the Xantusia and

Lepidophyma; night lizards in caves; island gigantism;

the species concept; the idea of unisexuals; night lizard

ecology; are night lizards nocturnal?; reproduction; so-

ciobiology; diet and predators; helminth parasites; ther-

mal and water ecology; movement, home range, and

population density; and conservation status.

The last half of the book provides a detailed discussion

about the night lizard family, Xantusiidae, followed by

the night lizard species accounts. Originally, the family

only had one species, Xantusia vigilis (Fig. 1), the small

lizard Janos Xantus collected at Fort Tejon, California.

But, eventually two other genera were added, Cricosaura

(Cuban night lizards), and Lepidophyma (tropical night

lizards). Bezy provides a detailed map that shows the dis-

tribution of the three genera and a diagram showing the

phylogenetic relationships. From there, each species has

its own detailed account, which generally includes these

sections: identification; chromosomes; size; distribution

and habitat; life history; sex ratio; etymology; conserva-

tion status; and discussion. Each account has a color pho-

to of the lizard, a colored distribution map, and photos

of representative habitat. With nearly 250 total figures

and photos throughout the book, the reader is treated to

a photo library that is unbeatable. Following the species

accounts is a night lizard species key—complete with

diagrams and photos. Also included are scale features for

differentiating night lizard species. The book ends with a

literature cited section which likely includes all the key

papers ever written on night lizards.

Overall, Bezy’s book is a must read for anyone inter-

ested in the story behind the night lizard, or in tales of

herpetological discovery and adventure in general. The

storytelling alone is reason enough to buy the book; the

photos, species accounts, range maps, etc., are a super

bonus and make the book the best resource currently on

this topic.

Howard O. Clark, Jr. has more than 20 years of professional wildlife and research experience. Howard

is certified by the Ecological Society of America as an ecologist and by The Wildlife Society (TWS) as a

Certified Wildlife Biologist®. His work as an ecological consultant has focused on the fauna and ecosystems

of California and has included extensive baseline inventories, surveys for rare animals, and habitat

assessments. He has conducted dozens of inventories, surveys, and assessments for Blunt-nosed Leopard

Lizard, Western Burrowing Owl, San Joaquin Kit Fox, Giant Kangaroo Rat, and Mohave Ground Squirrel

among many others. Howard developed his consulting skills while working for H. T. Harvey & Associates

(Los Gatos, California) for 10 years and Garcia and Associates (Auburn, California) for three years. He

currently works for Colibri Ecological Consulting, LLC, as a Senior Scientist in Fresno, California. Prior

to working as a consultant, Howard spent seven years as a wildlife biologist with the Endangered Species

Recovery Program (California State University, Stanislaus Foundation, Turlock, California). He completed

his Master’s degree at CSU, Fresno in 2001. His thesis addressed the interactions between the endangered

San Joaquin Kit Fox and the non-native Red Fox in Kern County, California. Howard is an instructor for

TWS kit fox and small mammal workshops, and the Western Section of TWS awarded him the Raymond

F. Dasmann Award for Professional of the Year in 2015. He is the Layout Editor for the Western Section’s

journal, Western Wildlife, as well as three herpetological journals: Amphibian & Reptile Conservation,

Sonoran Herpetologist, and Herpetological Conservation and Biology. During leisure time, Howard enjoys

hiking, geocaching, and visiting places of historical interest with his daughter.

Amphib. Reptile Conserv.

December 2019 | Volume 13 | Number 2 | e211

Official journal website:

amphibian-reptile-conservation.org

Amphibian & Reptile Conservation

13(2) [General Section]: 267-275 (e212).

Impacts of a highway on the population genetic structure of a

threatened freshwater turtle (G/lyptemys insculpta)

12.*Alexander J. Robillard, ?Sean Robinson, 7Elizabeth Bastiaans, and 7Donna Vogler

'Chesapeake Biological Laboratory, University of Maryland Center for Environmental Science, Solomons, Maryland, USA *Department of Biology,

State University of New York College at Oneonta, Oneonta, New York, USA

Abstract.—Genetic partitions for members of the family Emydidae often correspond with both natural and

anthropogenic landforms. For semi-terrestrial turtles, clear negative impacts are associated with habitat

fragmentation via roadways, such as loss of breeding individuals, increased inbreeding, and decreased migration.

The Wood Turtle (Glyptemys insculpta) is a Species of Special Concern in New York and native to the central

portion of the state, where Interstate Highway 88 was constructed in the 1970s. To examine possible impacts of

the highway on local populations, a museum collection of Wood Turtles that predates road construction was used.

Specifically, microsatellite markers were used to compare historic (n = 38) and contemporary (n = 26) Wood Turtle

DNA from opposite sides of the highway. The measured parameters were inbreeding (F,.), differentiation (F.,),

number of breeding individuals (N.), migration (m), and overall population genetic structure. The populations on

either side of the highway were predicted to have become more differentiated and inbred over time, and migration

was predicted to decrease over time. Overall, populations on either side of the interstate were historically a single

population, had a greater number of breeding individuals, and were less differentiated. No change in inbreeding

was found across time. These findings suggest there is more migration, running north to south between the two

populations, likely attributable to the directionality of the flow associated with local creeks. Further research

examining these two separate populations within the context of the entire state is necessary to determine whether

they should be treated as separate Conservation Units.

Keywords. Emydidae, habitat fragmentation, roadways, microsatellite, population structure

Citation: Robillard AJ, Robinson S, Bastiaans E, Vogler D. 2019. Impacts of a highway on the population genetic structure of a threatened freshwater

turtle (Glyptemys insculpta). Amphibian & Reptile Conservation 13(2) [General Section]: 267-275 (e212).

tion 4.0 International (CC BY 4.0): https://creativecommons.org/licenses/by/4.0/], which permits unrestricted use, distribution, and reproduction in any

medium, provided the original author and source are credited. The official and authorized publication credit sources, which will be duly enforced, are

as follows: official journal title Amphibian & Reptile Conservation; official journal website: amphibian-reptile-conservation.org.

Received: 22 February 2018; Accepted: 24 April 2019; Published: 23 December 2019

Introduction al. 2000). In North America, several case studies have

suggested that anthropogenic disturbances, particularly

On a global scale, amphibians and reptiles are in

decline due to pressures which include climate

change, unsustainable harvest, habitat loss, and habitat

degradation (Gibbons et al. 2000). Among all reptile and

amphibian species, members of the order Testudines are

particularly vulnerable to increased decline when faced

with increasing anthropogenic disturbances, such as road

mortality and illegal harvesting (Lieberman 1994; Garber

and Burger 1995; Wood and Herlands 1997; Williams

1999: Levell 2000; Gibbons et al. 2000; Gibbs and

Shriver 2002; Steen and Gibbs 2004; Gibbs and Steen

2005; Steen et al. 2006; USFWS 2015). Nearly half of

all turtle species are currently categorized as threatened

or endangered (Rhodin et al. 2011). Terrestrial turtles

are generally perceived as poor long-distance dispersers.

Limitations to dispersal enable habitat fragmentation,

which can put populations at risk of extinction due to

demographic and genetic diversity loss (Gibbons et

Correspondence. “ajrobill@umd.edu

Amphib. Reptile Conserv.

roadways, have direct negative impacts on freshwater

turtles by skewing sex ratios and increasing the mortality

of migrating individuals (Buhlman and Gibbons 1997;

Wood and Herlands 1997; Williams 1999; Levell 2000;

Gibbons et al. 2000; Gibbs and Shriver 2002; Steen and

Gibbs 2004; Gibbs and Steen 2005; Steen et al. 2006;

USFWS 2015).

When assessing the negative impacts of fragmented

populations, genetic markers can identify dispersal

pathways and population diversity (Lamb et al. 1989;

Galbraith et al. 1995). For example, studies focusing

on turtle populations have found river drainages and

intermontane basins to be barriers to gene flow (Gibbs

and Amato 2000). Similarly, several studies have

identified relatively high allelic diversity in Wood

Turtle populations when compared to other species

(Gibbs 1993; Tessier et al. 2005; Amato et al. 2008;

Castellano et al. 2009; Spradling et al. 2010). Given

December 2019 | Volume 13 | Number 2 | e212

Population genetics of Glyptemys insculpta in New York

Otsego (North) an d Delaware (South)

Counthes bisected by Interstate £3 and

the Susquehanna River

Fig. 1. Study area. Interstate Highway 88 (I-88) and the Susquehanna River (Susq.) bisect Otsego and Delaware Counties, New

York, USA.

their poor long-distance dispersal ability, populations of

freshwater turtles could be at increased risk from habitat

fragmentation via natural or anthropogenic barriers,

which may result in loss of genetic and demographic

connectivity (Gibbons et al. 2000; Gibbs and Amato

2000). Specifically, turtles which do attempt to move

long distances within a fragmented landscape, as females

often do for nesting, may be at greater risk of dispersal

related mortality, e.g., roadkill (Gibbs and Shriver 2002:

Steen and Gibbs 2004). Road mortality driven by habitat

fragmentation is believed to be the culprit for the gradual

skewing of sex ratios and the general decline of female

turtles among many freshwater turtle species throughout

the United States (Steen and Gibbs 2004; Gibbs and

Steen 2005; Steen et al. 2006).

Known to disperse both long and short distances

(Harding and Bloomer 1979) throughout its range, the

Wood Turtle (G/yptemys insculpta) is a Species of Special

Concern in New York State, which may be at risk from

the demographic and genetic impediments associated

with habitat fragmentation (Gibbons et al. 2000; Breisch

and Behler 2002; Tuttle and Carroll 2003; Arvisais et al.

2004; Sweeten 2008). Limitations to dispersal within

fragmented landscapes are believed to contribute to

the decline of wetland-dependent turtles (Gibbs 1993;

Tessier et al. 2005; Amato et al. 2008; Castellano et al.

2009; Spradling et al. 2010). Although previous studies

have examined Wood Turtle population genetics, none

have focused on the potential impacts that anthropogenic

Amphib. Reptile Conserv.

habitat fragmentation could have on regional populations

(Tessier et al. 2005; Amato et al. 2008; Castellano et al.

2009; Spradling et al. 2010).

Between 1974 and 1980, Interstate Highway 88 (I-

88, 193 km) was built across eight Central New York

counties (Fig. 1), including Otsego and Delaware

counties (Associated Press 1986; Edwardsen 1989).

Prior to this, from 1958-1968, Dr. John New collected

and dry-preserved Wood Turtles (n = 300) from across

New York state, including sites north and south of

I-88. Using this historic data set in conjunction with

contemporary data, this study examines the potential

impacts of building a large interstate highway on a

vulnerable turtle species over a 60-year period. Over

that same 60-year period, New York State has become

more populated, according to U.S Bureau of Census

data for 1960-2010. Given that wetland habitat size

and connectivity degrade with increased human activity

(Gibbs 2000), and that such disruption of wetland

mosaics can have dramatic negative impacts on semi-

terrestrial turtle populations (Gibbs and Shriver 2002),

declines are expected to be observable on the genetic

scale. Specifically, an increase in genetic differentiation

between populations (F,,), a decrease in the effective

breeding population size (N,) between sampling sites

on either side of Interstate 88, and limited gene flow

between populations on either side of the highway are

expected. Here, microsatellite data are used to examine

these critical genetic parameters.

December 2019 | Volume 13 | Number 2 | e212

Robillard et al.

Materials and Methods

Study sites: Contemporary (n = 26) and historic (n

= 38) data were collected from two streams in Otsego

County and two streams in Delaware County, New York.

The furthest sections of the two Otsego County streams

sampled are rectilinearly 11 km and 28 km north of

Interstate 88. The furthest sections of the two Delaware

county streams sampled are rectilinearly 3 km and 9 km

south of Interstate 88. All streams sampled are part of the

Susquehanna watershed, and terminate on the southern

side of Interstate 88 (Fig. 1). Contemporary sites were

sampled during the spring/early summer and late fall

active periods of 2015 and 2016 using the Regional

Conservation Needs protocol, which involves sampling

in 1 mi increments (Jones et al. 2015).

Samples: Blood samples were used for contemporary

data, and | mm tail tips were harvested from dried

historic specimens. Blood samples (0.1—0.5 ml) were

collected from the dorsal coccygeal vein using a sterile

1.0 ml 25-gauge syringe (Jones et al. 2015). Blood

was transferred into test tubes immediately upon

return from the field and stored in 1:1 1 x PBS buffer

in a -20 °C freezer. Genomic DNA was extracted from

each sample using the QIAGEN DNeasy Blood and

Tissue Kit (Qiagen, Inc., Valencia, California, USA).

Tail tips were digested in Proteinase K for 36 h. Each

extracted sample was stored in a -20 °C freezer. Seven

microsatellite loci were examined (GmuD16 [Genbank

accession number: AF516235], GmuD40 [AF517244],

GmuD51 [AF517239], GmuD87 [AF517244], GmuD88

[AF517245], GmuD93 [AF517248], and GmuD95

[AF517249]) using primers initially designed for a close

relative of the Wood Turtle, Glyptemys muhlenbergii

(King and Julian 2004). Samples were amplified

using the QIAGEN Multiplex PCR kit (Qiagen, Inc.,

Valencia, California, USA) and a modified version of

the PCR protocol (Castellano et al. 2009). The length

of the extension step from this protocol was doubled to

optimize historic sample amplification due to the highly-

fragmented nature of this DNA. PCR products were

analyzed at the Cornell Biotech Institute in Ithaca, New

York, and visualized using GENEMARKER version

2.6.7 (Hulce et al. 2011).

Statistical analysis: MICROCHECKER version 2.2.3

was used to test each locus for the presence of null alleles,

scoring errors, and large allele dropout (Van Oosterhout

et al. 2004). Clustering was used to assign individuals

to populations using the program STRUCTURE version

2.2 (Pritchard et al. 2000; Falush et al. 2003). Data

were analyzed for all contemporary and historic turtles

from each of the sites north (Otsego County) and south

(Delaware County) of Interstate 88. For each analysis,

three runs were used for each value of K (number of

assumed populations) ranging between one and nine.

Amphib. Reptile Conserv.

A 106 burn-in period was used, and 106 Markov Chain

Monte Carlo (MCMC) iterations were used in the default

“admixture model” of ancestry and correlated allele

frequencies. Population origin data (north and south)

were provided for each individual. Mean log likelihood

and DK values were used to assign individuals to

populations [K] (Evanno et al. 2005).

Deviations from the Hardy-Weinberg (HWE)

expectation among pairs of loci were tested along

with mean heterozygosity, allelic richness, numbers

of private alleles, inbreeding coefficient (F,.), genetic

differentiation (F..), and effective population size (N,)

using GenAIEx version 6.5 (Peakall and Smouse 2006,

2012) for the populations north (Otsego) and south

(Delaware) of Interstate 88. The comparison of historic

and contemporary loci F,. was made using a student’s

t-test. Evidence of a bottleneck on the contemporary

data was tested using the program BOTTLENECK

(version 1.2.02, Cornuet and Luikart 1996) with an

infinite allele model (IAM) and the two-phase model

(TPM) recommended by Luikart et al. (1998) over

10,000 iterations. The significance of Wilcoxon test

score output (a = 0.05) and mode shift were both used

as evidence of a bottleneck (Cornuet and Luikart 1996;

Luikart et al. 1998; Chiucchi and Gibbs 2010). Short-

term migration (m) between the “last few generations”

was estimated using Bayesian inference software

BAYESASS (version 3.03, Wilson and Rannala

2003) using 3 x 107 iterations with two long-chains

sampling every 2,000 iterations, and this included

a burn-in of 107. Specifically, a time span reaching

back < 5 generations, or 25-125 years (Chuicchi and

Gibbs 2010) was used since the estimated generation

time for Wood Turtles 1s 25 years (Farrell and Graham

1991; Galois and Bonin 1999). Multiple independent

model runs were made using a random seed, with final

selection made based on Maximum Likelihood (e.g.,

Chuicchi and Gibbs 2010).

Results

Six of the seven microsatellites amplified consistently.

The exception was GmuD51, which was removed

from further analyses. Historic specimens had a high

allele dropout rate (46%), which is expected for highly

fragmented antique DNA (Mills et al. 2000; Sefc et

al. 2003). No evidence of genotyping error or null

alleles was found for those that amplified. Sample size,

effective population size, heterozygosity, and overall

differentiation between north and south populations, for

both historic and contemporary data, are summarized

in Table 1. F,, values for the contemporary populations

was 0.166, while historically they were estimated at

0.081 (Table 2). Heterozygosity estimates per locus are

summarized in Table 2. Northern contemporary and

historic data each showed three of six loci out of HWE.

Southern contemporary data displayed four of six loci

December 2019 | Volume 13 | Number 2 | e212

Population genetics of Glyptemys insculpta in New York

North

0.869

(SD: 0.054)

Confidence Interval

(0.761 - 0.969)

a=I11

——-

m = 0.044 (SD: 0.040)

Confidence Interval

(0001 - 0.150)

ai = 0.241 (SD: 0.037)

Confideoce Interval

(0.166 - 0.302)

aaa

South

0.707

(SD: 0.034)

Confidence Interval

(0.668 - 0.786)

n= 15

Fig. 2. Estimate of short-term gene flow among populations

north and south of Interstate Highway 88 (gray bar) and the

Susquehanna River (dashed line) shown with 95% confidence

intervals. Circle size reflects relative sample size. Values inside

of circles represent the contribution of gene flow from within

populations.

out of HWE, while the southern historic data displayed

a single locus out of HWE. Comparison of historic and

contemporary inbreeding (F,.) indicated no difference

between estimations (P = 0.30). Fixation indexes for

each locus are summarized in Table 3.

Contemporary samples clustered into two populations

(K = 2, Ln P(D) = -659.5, Var [LnP(D)] = 74.5), with

a clear distinction between north and south samples.

Historic samples consistently clustered into a single

population (K = 1, LnP(D) = -787.9, Var [LnP(D)]

= 33.0). There was a deficiency of heterozygosity (P

0.04) for the northern contemporary population

under the Wilcoxon rank sign test in the TPM model.

Specifically, none of the northern loci were in mutation-

drift equilibrium, as five of six loci showed signs

of heterozygosity deficiency with the final locus in

excess under the IAM model. Southern contemporary

populations showed no sign of a genetic bottleneck.

Short-term migration (m) was conservatively estimated

to be higher going from north to south (24%) than south

to north (4.4%) [Fig. 2].

Discussion

Genetic changes and trends in turtle populations may be

difficult to detect due to their naturally long generation

times and long lives (Gibbs and Amato 2000). Using

the genetic material available from historic samples

allowed the successful detection of changes between

the populations across a relatively brief period of

time. Although this study used a limited number of

microsatellites (six of seven), the polymorphic nature

of the markers and highly differentiated level of the

populations suggest that the results capture an adequate

amount of information for comparative genetic analysis

(Kalinowski 2002, 2005; Arthofer et al. 2018), especially

given the sample sizes for the historic and contemporary

populations (Hale et al. 2012). Specifically, the results

indicated that these local populations have likely become

genetically fragmented over the last 60 years. This may

indicate that certain freshwater turtle populations are

more vulnerable, in terms of the rate of change, to shifts

in genetic structure than previously thought.

Structural analysis of the contemporary data revealed

that Wood Turtle populations clustered into two

populations, where historically, they were likely a single

interconnected unit. In addition, the same samples revealed

that local populations have become more differentiated

over time as an increase of F , was observed from 0.081

in historical to 0.166 in contemporary populations. This

shift from moderate differentiation (> 0.05) to great

differentiation (> 0.1) over an evolutionarily short period

of time would appear to be aberrant when compared to

previous studies examining Wood Turtle differentiation

(Hartl and Clark 1997). However, Tessier et al. (2005)

sampled Wood Turtles in a similar semi-montane habitat,

and found that some of their populations, which had a

proximity between sites comparable to those in the

current study (~15—50 km), had similar differentiation

as the historic samples studied here. Conversely, the

contemporary sample differentiation found here is more

similar to that of their sites which were much further apart

Table 1. Summarized outputs of population parameters from GenAIEx v 6.5. Parameters displayed are sample size (n), effective

population size (N.), observed (H,) and expected (H,) heterozygosity, and overall differentiation (F,,) between North and South

collecting sites.

Historic (1955-1965)

n N, (SE) Mean # alleles (SE)

North 20 6.9 (1.5) 10.0 (1.5)

South 18 513°C.) 7.8 (1.6)

Total 38 122

Contemporary (2015-2016)

North 11 4.0 (0.4) 6.3 (0.5)

South 15 7.0 (0.8) 11.0 (0.8)

Total 26

Amphib. Reptile Conserv. 270

H, (SE) H, (SE) Private alleles soe

0.61 (0.11) 0.82 (0.04) 40

0.65 (0.14) 0.77 (0.04) 27 0.081

0.67 (0.04) 0.74 (0.02) 12

0.81 (0.04) 0.85 (0.02) 19 0.166

December 2019 | Volume 13 | Number 2 | e212

Robillard et al.

Table 2. Summarized contemporary observed (H,) and expected (H,) heterozygosity.

Locus Size range (bp) # of alleles

GmuD16 165-296 24

GmuD87 238-394 24

GmuD88 114-264 30

GmuD93 125-389 20

GmuD95 122-266 24

GmuD40 157-280 21

(> 60 km) than the sites in this analysis (Tessier et al.

2005). Overall, this suggests the populations examined

here, which are relatively close to one another in terms

of physical distance, possess genetic differentiation

that is more akin to areas further apart, implying that

there is a barrier preventing genetic exchange between

them. In flatter areas along the coastal plains of the

northeastern United States, differentiation among Wood

Turtle populations is essentially non-existent, enabling

populations to be panmictic across separation distances

greater than 40 km (Castellano et al. 2009). This leads

us to believe that dispersal limitation is due to some

environmental factor, and not simply life history.

A study on another terrestrial emydid turtle, Jerrapene

ornata (Ornate Box Turtle), in Texas found that a major

highway built in 1937 was likely the cause of significant

differentiation between populations on either side, but not

the cause of a change in overall structure (Richtsmeier

et al. 2008; Cureton et al. 2014). Similarly, Tessier et

al. (2005) found that the St. Lawrence River acts as a

barrier between Wood Turtles on either side of its shores,

separating them structurally. It is possible that the

observed structural separation between the populations

studied here has been compounded by the combination

of the intertwining bisection of Interstate 88, and the

Susquehanna River (Fig. 1).

Despite the findings of Tessier et al. (2005), the

migration analysis output in this study (Fig. 2) may

suggest that flooding events are allowing at least

some unidirectional gene flow to persist between the

populations on either side of Interstate 88. In short, local

hydrology from lower order streams at both the north

North South

H, H, H, H,

0.700 0.830 0.857 0.829

0.700 0.840 0.857 0.768

0.733 0.849 0.867 0.838

0.333 0.736 0.714 0.885

0.778 0.833 0.667 0.871

0.778 0.747 0.917 0.892

and south sampling sites terminate at the Susquehanna

River south of Interstate 88. Research by Jones and

Sievert (2009) indicates that flooding events, which have

dramatic impacts on Wood Turtles, may play a vital role

in connectivity. Specifically, flood events can displace a

substantial (40%) portion of Wood Turtle subpopulations

by long distances (1.4—-16.8 km) downstream (Jones and

Sievert 2009), which may explain the unidirectionality of

the migration estimates found here (Fig. 2).

This possibility seems even more likely when

considering the local footprint of Interstate 88, much of

which is built on steep and sometimes craggy mounds

protruding from the stream and forest surface. These

mounds, which span four total lanes and occasionally

split with a center depression or open fall at the median

strip, most likely make terrestrial genetic exchange near

impossible between the north and south populations.

Moreover, the Susquehanna in its entirety is substantially

narrower than the St. Lawrence River (Kammerer 2005),

which may make survival of flooding events more

likely. Additionally, we observed Wood Turtles using the

Susquehanna’s embankments and flood plains with some

regularity, so although its flow may prevent and influence

movement, it should not be considered an insurmountable

genetic barrier like the much larger St. Lawrence (Tessier

et al. 2005). As such, it is likely that Wood Turtle

movement is influenced by the directionality of the flow

in creeks and rivers, and may explain the unidirectional

migration observed here (Fig. 2).

Although some streams and creeks in Delaware

County run north to south as they percolate down from

the Catskill mountains, the northern Delaware drainages

Table 3. Microsatellite fixation indexes for both historic and contemporary populations.

Historic

Locus K ",

GmU16 0.245 0.308

GmU87 -0.246 -0.045

GmU88 0.253 0.282

GmU93 0.956 0.961

GmU95 0.136 0.171

GmU40 -0.028 0.024

Mean 0.220 0.284

SE 0.166 0.147

Amphib. Reptile Conserv.

Contemporary

st F, it st

0.084 0.093 0.226 0.146

0.161 -0.056 0.102 0.150

0.038 0.030 0.157 0.131

0.110 0.117 0.367 0.283

0.040 0.166 0.270 0.125

0.051 -0.048 0.117 0.158

0.081 0.050 0.206 0.166

0.020 0.037 0.041 0.024

271 December 2019 | Volume 13 | Number 2 | e212

Population genetics of Glyptemys insculpta in New York

near the sampling sites used in this study flow south

to north, terminating into the Susquehanna. Previous

research by Brown et al. (2016) indicated that Wood

Turtles become more terrestrial as the thermoregulatory

benefits of returning to the water at night diminish

during the summer, but they never seem to stray too

far from flowing water. Furthermore, a species of turtle

that divides its time between land and water (Kaufmann

1992) is expected to use smaller rivers as corridors like

many other turtle species (Gibbs and Amato 2000). If

local Wood Turtles are using the Susquehanna River as

a corridor at least in part, with strong flow and flooding

events acting as a migration regulator, further genetic

research should yield an F., gradient and not complete

differentiation. In other words, central New York’s

populations should be progressively more differentiated

from populations further south along the Susquehanna

River, but not completely differentiated altogether.

Therefore, further research should investigate the

potential of large rivers, namely the Susquehanna River,

to act as turtle barriers or corridors.

If the Susquehanna is acting as a unidirectional

barrier, long-term declines could prove problematic for

the local populations. Specifically, the loss of only a few

individuals may appear to be minimal in terms of allelic

diversity, but a negative change in the effective breeding

population (N,), as was observed, could prove to have

adverse conservation consequences. For example, similar

rates of reduction in Bog Turtle populations have been

identified as substantially increasing the likelihood of

extirpation (Shoemaker 2011). Certain life histories are

also known to be susceptible to such impacts (Jonsson and

Ebenman 2001). Specifically, for Bog Turtle populations,

the loss of only a few breeding adult individuals can have

greater impacts on populations than even dramatic short-

term increases in juvenile mortality (Shoemaker 2011).

Similar losses for Wood Turtles, the closest known

relative of the Bog Turtle, could prove to be equally

problematic. This vulnerability, again, would be due to

their long generation times, high juvenile mortality rate,

and reliance on adult survival to bolster the populations

(Gibbs and Amato 2000). This situation presents a suite

of unique conservation issues which are likely to also be

applicable to other freshwater turtle species.

One noteworthy observation here is that the bottleneck

analysis presented identified both heterozygosity

deficiency and excess in the northern populations.

Typically, heterozygosity deficiency is associated with

a founder effect (Cornuet and Luikart 1996) or possibly

the existence of a subpopulation structure within the

sample, known as the Wahlund effect (Wahlund 1928).

However, in rare situations when allelic diversity is high,

as itis with the Wood Turtles in this study, heterozygosity

deficiency can be the result of post-bottleneck changes,

such as mutation or population expansion, which fill the

allelic gaps left by limited random selection (Cornuet and

Amphib. Reptile Conserv.

Luikart 1996; Maruyama and Fuerst 1985). For freshwater

turtle populations, which typically have a small number

of long-lived adults possessing the majority of effective

alleles (Crouse and Frazer 1995; Gibbs and Amato 2000),

the loss and sequential replacement of these few valuable

reproductive individuals appear to enable this particular

scenario. Considering that none of the loci for the

northern population were in mutation-drift equilibrium,

and they showed evidence of deficiency and excess, it

seems clear that something has impacted the northern

population allelic ratios. Further research into Otsego

County’s populations north of Interstate 88 is required

to determine the source of this irregularity. Additionally,

we recommend that future management plans for Wood

Turtle populations in central New York and other regions

with montane-riverine mosaics consider the potential

genetic complications associated with anthropogenic

habitat fragmentation. To mitigate these potential

impacts, the installation of appropriately sized culverts,

drift nets, and turtle-crossing signs (Aresco 2005; Woltz

et al. 2008), in high-density areas (Gunson and Schueler

2012), is necessary.

Conclusions

Consistent with other research (Steen and Gibbs 2004;

Gibbs and Steen 2005; Steen et al. 2006) the observed

division and reduction in N, in the local populations

studied here, the potential recent genetic bottleneck,

the increased differentiation, and the overall change in

population structure are most likely attributable to the

additional fragmentation of the local montane/riverine

habitat by the bisecting interstate highway. A clear

north to south directionality of gene flow was observed

from the short-term (25—125 years) migration estimate.

The full implications of this dichotomy, in the context

of potential isolation due to fragmentation, have yet to

be determined. As a species of conservation concern,

understanding the genetic landscape at the local and

regional levels is vital for planning future management.

In terms of conservation, it is possible New York’s central

populations hold unique alleles as they are surrounded by

three major highways and two large mountain ranges. In

turn, this may require that management efforts treat these

isolated populations as demographically independent

units, should they yield unique genetic variation. As

such, we recommend that policy and management

reflect the impacts that bisecting highways can have

on populations within a local region, and not just those

adjacent to a thoroughfare. Additionally, we recommend

that policy and management efforts reflect the evidence,

which suggests that hydrology may dictate Wood Turtle

gene flow. Furthermore, research focused on determining

where central New York’s populations fit within the

context of the entire region’s genetic landscape will be

particularly useful.

December 2019 | Volume 13 | Number 2 | e212

Robillard et al.

Acknowledgments.—This research was funded by

the State University of New York College at Oneonta

Sponsored Programs research grant, the Western New

York Herpetological Society’s Marv Aures and Bob

Krantz research grant, the Huyck Preserve research grant,

and the State Wildlife Grant (SWG) Program. Special

thanks to E. Clifton, S. Fontaine, W. O’Connell, and S.R.

Talley for providing field and lab assistance. Wood Turtle

handling and tissue collection was permitted by the New

York State Department of Environmental Conservation

(NYSDEC; Science Permit #139) and was approved by

the SUNY-Oneonta Institutional Animal Care and Use

Committee. We are appreciative of the initiated fieldwork

and consultation of Dr. Tom Akre, Dr. Glenn Johnson,

Dr. Mike Jones, Dr. Angelena Ross, Bill Hoffman, and

Michael Musnick.

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Alex Robillard received a B.S. in Conservation Biology from State University of New York, College of

Environmental Science and Forestry (SUNY-ESF, Syracuse, New York, USA), and an MLS. in Biology

from SUNY-Oneonta. Currently a predoctoral fellow at the Smithsonian Data Science Lab and National

Zoo, and a Ph.D. student in the Marine-Estuarine and Environmental Science Program at the University

of Maryland, Alex’s dissertation focus is on the conservation and population genetics of the Eastern

Pacific Hawksbill Sea Turtle. His past research had focused on the ecology of the Bog Turtle, Wood

Turtle, and Eastern Massasauga Rattlesnake in New York State. Alex is also researching how deep

machine learning and computer vision can be used to combat the poaching of sea turtles.

Sean Robinson completed a B.A. at Hartwick College (Oneonta, New York, USA), an M.S. at SUNY-

ESF, and a Ph.D. at the University at Albany, New York. Dr. Robinson joined the SUN Y-Oneonta

faculty in 2010 where his research is focused on understanding how the mode of reproduction in plants,

particularly bryophytes, affects colonization of new habitats, range expansions, and the exchange of

alleles both within and between populations. Additionally, Dr. Robinson conducts research focused on

vegetation dynamics on alpine summits, using molecular techniques to identify population structuring.

Elizabeth Bastiaans completed a B.A. at the University of Chicago, a Ph.D. at the University of

California, Santa Cruz, and a postdoctoral fellowship at the University of Minnesota, Twin Cities.

Elizabeth joined the SUNY-Oneonta faculty in 2015. Her previous research focused on sexual signal

evolution in Mexican montane lizards and life history evolution in tropical crickets. At SUN Y-Oneonta,

Dr. Bastiaans has started to focus her research on the reproduction and physiology of the Wood Turtle

across New York State, while maintaining her previous collaborations with colleagues in Mexico.

Donna Vogler was born and educated in the Midwestern United States, with a B.S. from The Ohio

State University, and an M.S. from Iowa State University, before working for the U.S. Fish and Wildlife

Service in Washington, DC. Donna earned a Ph.D. from Penn State University in Botany, and was a post-

doctoral researcher at the University of Pittsburgh before joining the SUNY-Oneonta faculty in 2000. Dr.

Vogler’s recent research topics include demographic studies of invasive plant species (e.g., Marsh Thistle,

Cirsium palustre), floral mechanisms related to self vs. outcross pollination, and using Wood Turtle habitat

communities and vegetation management at regional airports to reduce wildlife hazards.

Amphib. Reptile Conserv.

December 2019 | Volume 13 | Number 2 | e212

Official journal website:

amphibian-reptile-conservation.org

Amphibian & Reptile Conservation

13(2) [General Section]: 276-298 (e213).

Modelling the distribution of the Ocellated Lizard in France:

implications for conservation

1*Pierre Jorcin, 7Laurent Barthe, *Matthieu Berroneau, ‘Florian Dore, °Philippe Geniez, °Pierre

Grillet, “Benjamin Kabouche, *Alexandre Movia, °Babak Naimi, ‘°Gilles Pottier, ‘Jean-Marc Thirion,

and ‘*Marc Cheylan

'Naturalia-Environnement, Site Agroparc, rue Lawrence Durrell, 84911 Avignon, FRANCE *!°Nature En Occitanie, Maison régionale de

l’Environnement, 14 rue de Tivoli, 31000 Toulouse, FRANCE ?Cistude-Nature, Chemin du Moulinat, 33185 Le Haillan, FRANCE +3 Chemin de

Saint-Jacques, Faugerit, 79120, Chey, FRANCE »*'*Laboratoire Biogéographie et Ecologie des Vertébrés — CNRS, PSL Research University, EPHE,

UM, SupAgro, IRD, INRA, UMR 5175 CEFE, 1919 route de Mende, Montpellier, FRANCE °10 rue de la Sayette, 79340 Vasles, FRANCE ‘Ligue

pour la Protection des Oiseaux Provence-Alpes-Cote d’Azur (LPO PACA), 6 avenue Jean Jaurés, 83400 Hyéres, FRANCE *Ligue pour la Protection

des Oiseaux Dréme (LPO Droéme), 18 place Génissieu, 26120 Chabreuil, FRANCE °Department of Geosciences and Geography, University of

Helsinki, 00014, PO Box 64, Helsinki, FINLAND ''Objectifs Biodiversité, 22 rue du Dr. Gilbert, 17250 Pont-l’Abbé-d’Arnoult, FRANCE

Abstract—The Ocellated Lizard, Timon lepidus (Daudin 1802) occupies the Mediterranean regions of

southwestern Europe (Portugal, Spain, France, and the extreme northwest of Italy). Over the last decades, a

marked decline in its population has been observed, particularly on the northern edge of its distribution. As a

result, it is currently considered a threatened species, especially in France and Italy. In France, a national action

plan for its conservation has been put in place. In this study, ecological niche modelling (ENM) was carried out

over the entire area of France in order to evaluate the species’ potential distribution, more accurately define its

ecological niche, guide future surveys, and inform land use planning so this species can be better taken into

consideration. The modelling used data representing 2,757 observation points spread over the known range

of the species, and 34 ecogeographical variables (climate, topography, and vegetation cover) were evaluated.

After removing correlated variables, models were fitted with several combinations of variables using eight

species distribution model (SDM) algorithms, and then their performance was assessed using three model

accuracy metrics. Iterative trials changing the input variables were used to obtain the best model. The optimized

model included nine determining variables. The results indicate the presence of this species is linked primarily

to three climate variables: precipitation in the driest month, precipitation seasonality, and mean temperature

in the driest quarter. The model was checked by a sample dataset that was not used to fit the model, and this

validation dataset represented 25% of the overall field observations. Of the known occurrence locations kept

aside to check the results, 94% fell within the presence area predicted by the modelled map with a presence

probability greater than 0.7, and 90% fell within the area with a presence probability ranging from 0.8 to 1, which

represents a very high predictive value. These results indicate that the models closely matched the observed

distribution, suggesting a low impact of either geographical factors (barriers to dispersal), historical factors

(dispersal process), or ecological factors (e.g., competition, trophic resources). The overlap between the

predicted distribution and protected areas for this species reveals that less than 1% of the potential distribution

area is protected by strong regulatory measures (e.g., national parks and natural reserves). The knowledge

obtained in this study allows us to recommend some guidelines that would favor the conservation of this

species.

Résumé.—Le lezard ocellé, Timon lepidus (Daudin 1802), occupe les regions méditerranéennes du sud-ouest

de l’Europe (Portugal, Espagne, France, et extréme nord-ouest de I’Italie). Au cours des derniéres décennies,

un fort declin des populations a été observe, particulierement aux marges nord de sa distribution. Il est donc

considéreé comme une espece menacée, spécialement en France et en Italie. En France, il bénéficie d’un

plan national d’actions en faveur de sa preservation. La modélisation de sa distribution a ete conduite sur

l'ensemble du territoire national en vue d’estimer sa distribution potentielle, préciser sa niche écologique,

orienter les prospections futures et permettre une meilleure prise en compte de Il’espece dans l’aménagement

du territoire. Le travail de modelisation repose sur 2757 points d’observation répartis sur l’?ensemble de la

distribution connue de l’espéce, confrontes a 34 variables climatiques, topographiques, et de couvert vegetal.

Apres suppression des variables autocorrélées, plusieurs combinaisons de variables ont éte testees, et leur

performances évaluées a partir de huit algorithmes SDM. Le meilleur modele retient neuf variables, déterminees

par l’algorithme ayant la meilleure performance. Les modeles montrent que la presence de l’espéce est

Correspondence. ':*p.jorcin@naturalia-environnement.fr, ?1.barthe@natureo.org, * matthieu.berroneau@cistude.org,

*florian.dore@gmail.com, ° Philippe.geniez@cefe.cnrs.fr, ° p.grille(@wanadoo fr, ’ benjamin.kabouche@lpo.fr, *alexandre.movia@lpo.fr,

*naimi.b@gmail.com, '° g.pottier@natureo.org, |! thirion.jean-marc@sfrfr, '* marc.cheylan@cefe.cnrs.fr

Amphib. Reptile Conserv. 276 December 2019 | Volume 13 | Number 2 | e213

Jorcin et al.

principalement déterminée par la sécheresse et la température estivale: précipitations au cours du mois le

mois le plus sec, saisonnalite des précipitations et temperature moyenne des trois mois les plus chauds. La

validation du modele sur la base d’un échantillon totalisant 25 % du total des observations, non inclus dans le

modele, montre que 94 % des données de validation se placent dans I’aire potentielle au seuil de probabilite de

0,7, et 90 % pour une probabilité comprise entre 0,8 et 1. Ceci donne une valeur predictive tres elevee au modeéle

retenu. On constate une étroite concordance entre la distribution potentielle et la distribution réalisée, ce qui

suggere une faible influence des facteurs géographiques (obstacles a la dispersion), historiques (processus

de dispersion) ou ecologiques (competition, ressources trophiques, etc.). Le croisement cartographique entre

l’aire potentielle de ’espece et les espaces protéegés montre que moins de 1 % de Il’aire potentielle est couverte

par des mesures réglementaires fortes (parcs nationaux et reserves naturelles). En conclusion, le travail donne

des orientations pour ameéliorer la connaissance de la distribution de l’espece et des pistes de réflexion en

faveur de sa conservation.

Keywords. Timon lepidus, species distribution models, ecological niche, Europe, Reptilia, Sauria, Squamata

Citation: Jorcin P, Barthe L, Berroneau M, Doré F, Geniez P, Grillet P, Kabouche B, Movia A, Naimi B, Pottier G, Thirion J-M, Cheylan M. 2019.

Modelling the distribution of the Ocellated Lizard in France: implications for conservation. Amphibian & Reptile Conservation 13(2) [General Section]:

276-298 (e213).

4.0 International (CC BY 4.0): https://creativecommons.org/licenses/by/4.0/], which permits unrestricted use, distribution, and reproduction in any

medium, provided the original author and source are credited. The official and authorized publication credit sources, which will be duly enforced, are

as follows: official journal title Amphibian & Reptile Conservation; official journal website: amphibian-reptile-conservation.org.

Received: 11 December 2018; Accepted: 23 August 2019; Published: 22 December 2019

Introduction

The success of conservation programs in protecting

threatened species depends largely on the quality of

the information related to the environmental condi-

tions favorable to (or sought by) the species (Griffith et

al. 1989; Souter et al. 2007; Fourcade et al. 2018). For

this reason, ecological (or environmental) niche mod-

els (ENMs) [Sillero 2011], also known as species dis-

tribution models (SDMs) or habitat distribution models

(HDMs), are increasingly used to inform conservation

measures (Ferrier 2002; Graham et al. 2004; Araujo and

Segurado 2004; Santos et al. 2009; Elith and Leathwick

2009; Lyet et al. 2013; Jiang et al. 2014; Wan et al. 2016:

Tanella et al. 2018). These models allow researchers to

identify the factors which explain the distribution of a

species (Austin et al. 1990; Vetaas 2002; Guisan and

Hofer 2003; Jiang et al. 2014; Ferreira et al. 2013), to

orient research toward zones where the species has not

yet been identified (Engler 2004; Raxworthy et al. 2003;

Lyet et al. 2013; GHRA-LPO Rhone-Alpes 2015; Ryberg

et al. 2017), to identify the most favorable zones for the

conservation of the species (Brito et al. 1996; Barbosa

et al. 2003; Anderson and Martinez-Meyer 2004; Mufioz

et al. 2005; Guisan et al. 2013; Lyet et al. 2013; Wang

et al. 2016; Moradi et al. 2019; Sohrab et al. 2019), to

evaluate the potential dispersion and gene flow between

population patches (Guisan and Thuiller 2005), to model

future changes in distribution (for example, based on cli-

mate change) [Franklin 1998; Guisan and Hofer 2003;

Araujo et al. 2006; Carvalho et al. 2011; Cheaib et al.

2012; Ianella et al. 2018; Renwick et al. 2018], as well as

to project the distribution into past scenarios (Sillero and

Carretero 2013).

Species distribution models also allow comparisons

Amphib. Reptile Conserv.

between potential and actual distributions, two very use-

ful concepts to consider in conservation biology. Put sim-

ply (see Pulliam 2000 for a more detailed explanation),

a potential niche corresponds to areas in which the cli-

matic, terrain, and habitat conditions are theoretically fa-

vorable to the species in the current conditions, whereas

the realized niche takes into account historical and bi-

otic factors that may explain the absence of the species

within the ecological area defined by the fundamental

niche. Comparing these two niches provides informa-

tion about the historical processes that led to the cur-

rent distribution, the dispersion capacity of the species,

and the obstacles to its dispersion. This can illuminate

the ecological factors that negatively influence the pres-

ence of the species, such as the presence of competitors

or predators, unsuitable habitats, insufficient trophic re-

sources, a lack of host species, and others (see Guissan

and Thuiller 2005). When used to model the distribution

of a species in decline, SDMs can also provide informa-

tion about the causes of decline, particularly by helping

to differentiate the proportions due to global factors as

opposed to regional or local factors (Jiang et al. 2014). In

some cases, SDMs can even allow the extent of decline

to be measured, by comparing the potential niche with

the observed niche (Lyet et al. 2013; Ryberg et al. 2017).

The Ocellated Lizard, Timon lepidus (Daudin 1802),

is a good case study for this type of analysis. This species

occupies the Mediterranean regions of southwestern Eu-

rope (Portugal, Spain, France, and the extreme northwest

of Italy) [Figs. 1 and 2]. At the edges of its distribution,

it has faced a marked population decline over the last de-

cades and is now considered a threatened species, espe-

cially in France and Italy (Salvidio et al. 2004; Cheylan

and Grillet 2005; Cheylan 2016).

This species is closely linked to the Mediterranean

December 2019 | Volume 13 | Number 2 | e213

Distribution of Timon lepidus in France

Jean Nicolas.

climate and specific biotopes (Doré et al. 2015). More-

over, due to its large size and thermal requirements for

reproduction, it 1s particularly demanding regarding cli-

matic conditions (Mateo 2011). The incubation period

for its eggs is long, about 100 days, making use of the

full period of warmer temperatures. The eggs hatch late,

generally at the end of September or the beginning of

October (Bischoff et al. 1984; Doré et al. 2015). Hence,

two main factors drive the distribution of this species: its

ecophysiology (thermal requirements linked to the indi-

vidual’s bulk and to the incubation of the eggs) and its

habitat requirements (dry habitats with little tree cover).

In view of these needs, the Ocellated Lizard should theo-

retically benefit from the warming temperatures recorded

in Europe over the last 30 years (see Prodon et al. 2017).

Yet observations show a rather widespread decline of this

species, particularly on the northern edge of its distribu-

tion range, which is inconsistent with the expected situa-

tion in warming conditions (Salvidio et al. 2004; Cheylan

and Grillet 2005; Doré et al. 2015). This raises questions

regarding the causes of the decline of this species and, at

first glance, suggests a hypothesis that local effects pre-

dominate over global effects.

The use of SDMs allows the study of interesting bio-

geographical questions about this species. Native to the

Iberian Peninsula, the Ocellated Lizard colonized France

and the extreme west of Italy along the Mediterranean

coast (Sillero et al. 2014; Doré et al. 2015). This colo-

nization involved overcoming considerable physical ob-

stacles, including rivers, mountains, and forests. Niche

modelling can provide information about constraints that

limit dispersion; that 1s, whether the current distribution

boundaries of the species are of a climatic nature (thus

ecophysiological) or a physical nature (due to obstacles

to dispersion). The same question applies to the species’

colonization of the French Atlantic coast: Are the isolat-

ed populations along this coast the result of a process of

decline linked to the progressive degradation of habitat

or to climatic constraints? The responses to these ques-

tions can be found by comparing the expected distribu-

Amphib. Reptile Conserv.

Fig. 2. Timon lepidus, juvenile, Hérault, France. Photo by Jean

Nicolas.

tion with the observed distribution; that is, by comparing

the potential niche with the realized niche.

This study used species distribution modelling to 1n-

vestigate the following questions: (1) Does the observed

distribution of the Ocellated Lizard match its potential

distribution? If not, why not? (2) Which variables best

explain the distribution of this species: climate, terrain,

land use, or other factors? (3) Why is this species retreat-

ing at the edges of its distribution range, in contrast to

what might be expected based on climatic changes? (4)

Are the distribution boundaries of this species conditional

on either climate, physical barriers, or ecological causes?

(5) Which zones are potentially the most favorable for

the conservation of this species? (6) Where should future

surveys be carried out to improve our understanding of

the distribution range? (7) Based on these findings, what

conservation strategy should be implemented for the

conservation of the species?

Materials and Methods

Data

Ocellated Lizard dataset. This study used observations

(presence data only, Brotons et al. 2004) from an exhaus-

tive database that includes most of the occurrences ob-

served in France between 1970 and 2016. As the objec-

tive was essentially practical, 1.e., to identify the areas

of the potential presence of the species with the aim of

its conservation, this study did not consider taking into

account the entire distribution of the species as relevant

to building the model. This would have led to further

complications, such as the need to consider markedly di-

vergent genetic lines and, as a result, ecophysiological

adaptations specific to the regions that host these genetic

lines.

The data were collected by a number of organizations

and individuals over a period of 46 years. Before inte-

grating the data into the models, the records were verified

and cross-validated, keeping only precisely georefer-

December 2019 | Volume 13 | Number 2 | e213

Jorcin et al.

02550 100km

Fig. 3. Localization of the total presence data for the Ocellated Lizard Timon lepidus collected in Fran

2016.

enced locations (either data captured by GPS or observa-

tions recorded with a spatial positioning error of less than

100 m). This resulted in presence data for a total of 5,521

locations spread over southern France. From this, a sam-

ple dataset was extracted for modelling purposes. First,

only occurrences for the period that corresponded to the

vegetation variable used in the model were selected, tak-

ing into account the development and continuity of the

vegetation cover over the years. To fit this requirement,

the presence data were narrowed down to a sourcing pe-

riod of 16 years, which included 4,282 observations from

2000 to 2016. The distribution of occurrence data for this

period covers the whole area of study, though data prior

to the year 2000 were not included (Fig. 3).

Secondly, the dataset was filtered to avoid spatial bias

due to oversampling at particular locations, as field in-

vestigations conducted for environmental impact assess-

ments and other monitoring programs led to a higher con-

centration of data at certain sites. Therefore, the density

of points per km? was evaluated to identify zones subject

to sampling bias (Fig. 1 in Supplementary Materials),

using kernel density calculation. Through this analysis,

zones where the point density ranged from 2—50 points

per km? were determined, and for these zones, a single

record was retained per km?. After filtering the data by

density, a total of 2,757 occurrences were retained, pro-

viding presence data that was well distributed over the

study area (Fig. 4). Finally, of these 2,757 valid records,

75% were randomly selected for modelling, with the re-

maining 25% serving as an independent source to check

(validate) the results (Hirzel and Guisan 2002) [Fig. 4].

During the modelling procedure, the database consisting

Amphib. Reptile Conserv.

ane) a, HERE Dy

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@ Presence data collected 1970-1999 (1,239 points)

® Presence data collected 2000-2016 (4,282 points)

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ce during the period 1970 to

of 75% of the occurrences was itself divided into training

data (80%) and testing data (20%).

Environmental data. A set of ecogeographical variables

was used to model species distribution, taking into ac-

count the ecological requirements of the species (Guisan

and Thuiller 2005). A group of 34 variables was evalu-

ated through several iterations in order to identify and

include the most relevant variables (Table 1).

As a first step, environmental variables were obtained

from available sources at the appropriate spatial and the-

matic resolutions. This study used the CHELSA data-

base, version 1.1 (Climatologies at High Resolution for

the Earth’s Land Surface Areas, Karger et al. 2017). This

database includes a set of bioclimatic variables, with

monthly mean temperature and precipitation patterns,

for the time period 1979-2013, which corresponds to the

time range of the species occurrence data. The CHEL-

SA database is an alternative source to the widely used

WorldClim global climate database, as both derive their

bioclimatic variables from the monthly minimum, maxi-

mum, and mean temperatures, as well as precipitation

values. However, as described by Karger et al. (2017),

the CHELSA variables include additional corrections,

such as monthly mean and station bias, wind effect and

valley exposition, as well as correction for orographic

effects on precipitation. Additionally, as CHELSA is a

recently released product, this study allowed evalua-

tion of its potential for species distribution modelling.

In a recent study, Karger et al. (2017) highlighted differ-

ences observed at a large scale between WorldClim and

CHELSA models, with the latter leading to a significant

December 2019 | Volume 13 | Number 2 | e213

Distribution of Timon lepidus in France

‘ ¢

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Fig. 4. Localization of occurrence data for the Ocellated Lizard Timon lepidus, showing the points in the dataset used for the model

(red) and those used to check the model (blue).

improvement in SDM performance. The CHELSA bio-

clim data grid has a 30-arc-second pixel resolution, cor-

responding to a spatial resolution of 0.00833 decimal de-

grees at the equator. Applied to the study area in southern

France, the bioclimatic variables have a spatial resolution

of less than 1 km’, with each grid roughly covering an

area of 700 x 900 m.

As the CHELSA database does not include variables

reflecting solar radiation and its impact on climatic hu-

midity or aridity, data produced by the Consultative

Group on International Agricultural Research (CGIAR)

were used (Fick and Hiymans 2017). Data available at the

same resolution as the CHELSA dataset included mean

annual solar radiation, potential evapotranspiration, and

a global aridity index calculated from the ratio of mean

annual precipitation to mean annual potential evapo-

transpiration.

During the process of selecting and consolidating the

ecogeographical variables, another climate data source

was used to compare the results and identify the best input

that would optimize performance. This climate dataset

(Joly et al. 2010) is based on data from Meétéo France’s

weather stations and gathers a number of indicators that

are not provided in the CHELSA dataset, such as the

number of days with a temperature above 30°C or below

-5 °C, as well as other parameters related to temperature

ranges and seasonal variations over the year.

To increase model performance, topographic data

were also included. The elevation was obtained from the

EU-DEM v1.1 dataset, produced in the framework of

the European Commission’s Copernicus program (Bash-

field and Keim 2011), which includes a digital elevation

Amphib. Reptile Conserv.

model captured in 2011 and projected in ETRS89-LAEA

(EPSG code 3035), with a spatial resolution of 25 m.

This allowed the modelling to take altitude into account,

as well as slope and aspect, calculated in projected coor-

dinate systems (all in meters).

A variable representing vegetation cover is also re-

quired, in order to identify natural habitats suitable for

the Ocellated Lizard. The vegetation cover indicator se-

lected was the normalized difference vegetation index

(NDVI) generated from the MODIS (Moderate resolu-

tion Imaging Spectro-radiometer) Terra satellite sensors

(Huete et al. 2010), which has a spatial resolution of 250

m. Data captured in the first week of July were used, as

this corresponds to the optimal season for identifying

the permanent vegetation cover of interest in this study.

Vegetation indices in summer can highlight chlorophyll

produced by permanent vegetation, whereas vegetation

indices calculated in spring are influenced by the annual

growth of the herbaceous stratum.

Considering the possibility that land cover has

changed over time, the importance of changes in veg-

etation cover over the years was evaluated. The NDVI

values were compared for each year from 2000 to 2016,

calculating the variance and the standard deviation of

the mean NDVI value for this 16-year period (Fig. 5).

The results indicated that the change in permanent veg-

etation cover in the area of interest has been negligible,

especially in areas where the Ocellated Lizard has been

observed. Over 16 years, for the month of July, the stan-

dard deviation of the yearly NDVI values relative to the

NDVI mean value was below 0.075 for 98.9% of the en-

tire studied area, below 0.05 for 88.9% of the area, and

280 December 2019 | Volume 13 | Number 2 | e213

Jorcin et al.

Table 1. Description of the 34 variables evaluated.

Variable Description Source Period Resolution

Biol Annual mean temperature CHELSA v1.1 1979-2013 ~1 km?

Bio3 Isothermality (BIO2/BIO7) (* 100) CHELSA v1.1 1979-2013 ~1 km?’

Bio4 Temperature seasonality (standard deviation *100) CHELSA v1.1 1979-2013 ~1 km?

Bio5 Max temperature in warmest month CHELSA v1.1 1979-2013 ~1 km?’

Bios Mean temperature in wettest quarter CHELSA v1.1 1979-2013 ~1 km?

Bio9 Mean temperature in driest quarter CHELSA v1.1 1979-2013 ~1 km?’

Biol0 Mean temperature in warmest quarter CHELSA v1.1 1979-2013 ~1 km?

Biol3 Precipitation in wettest month CHELSA v1.1 1979-2013 ~1 km?

Biol4 Precipitation in driest month CHELSA v1.1 1979-2013 ~1 km?’

Biol5 Precipitation seasonality (coefficient of variation) CHELSA v1.1 1979-2013 ~1 km?

Biol6 Precipitation in wettest quarter CHELSA v1.1 1979-2013 ~1 km?’

Biol7 Precipitation in driest quarter CHELSA v1.1 1979-2013 ~1 km?

Bio 18 Precipitation in warmest quarter CHELSA v1.1 1979-2013 ~1 km?’

Biol9 Precipitation in coldest quarter CHELSA v1.1 1979-2013 ~1 km?’

EU-DEM v1.0 European

Alt Elevation Commission Copernicus program 2011 25m

Slope Slope Calculated from EU-DEM v1.0 2011 25m

Aspect Slope orientation Calculated from EU-DEM v1.0 2011 25m

NDVI Normalized vegetation index MODIS TERRA (NASA) July 2012 250m

TCD Tree cover density Copernicus 2012 20m

srad Mean annual solar radiation WORLDCLIM v.2 1970-2000 ~1 km?’

PET Global potential evapotranspiration CGIAR 1950-2000 ~1 km?

Global aridity index (mean annual precipitation / mean

Aridity annual PET) CGIAR 1950-2000 ~1 km?

TMO Annual mean temperature ThéMA - CNRS-UMR 6049 1971—2000 250m

TMN Number of days with temperature below -5 °C ThéMA - CNRS-UMR 6049 1971-2000 250m

TMX Number of days with temperature above 30 °C ThéMA - CNRS-UMR 6049 1971-2000 250 m

TAM Annual temperature range ThéMA - CNRS-UMR 6049 1971—2000 250m

TEH Inter-annual temperature variability in January ThéMA - CNRS-UMR 6049 1971—2000 250 m

TEE Inter-annual temperature variability in July ThéMA - CNRS-UMR 6049 1971—2000 250 m

Variance between January precipitation and monthly mean

PDH precipitation ThéMA - CNRS-UMR 6049 1971-2000 250 m

Variance between July precipitation and monthly mean

PDE precipitation ThéMA - CNRS-UMR 6049 1971-2000 250 m

PJH Number of rainy days in January ThéMA - CNRS-UMR 6049 1971—2000 250m

PEH Inter-annual precipitation variability in January ThéMA - CNRS-UMR 6049 1971—2000 250 m

PEE Inter-annual precipitation variability in July ThéMA - CNRS-UMR 6049 1971—2000 250 m

Variation between autumn (September and October) and

PRA July precipitation ThéMA - CNRS-UMR 6049 1971-2000 250 m

only reached a maximum of 0.2 for 1.1% of the area. An

assessment of the changes in vegetation cover within the

Methodology

study area allowed the selection of the NDVI mean value

for the 16-year NDVI dataset (from 2000 to 2016).

All the selected variables were aggregated to a spa-

tial resolution of ~1 km, using bilinear resampling tech-

niques.

Amphib. Reptile Conserv. 281

The performance of models was explored based on dif-

ferent combinations of ecogeographical variables using

an iterative approach (Heikkinen et al. 2006). To do this,

a model was first fitted with a selected set of variables

(Bucklin et al. 2014) and its performance was measured.

December 2019 | Volume 13 | Number 2 | e213

Distribution of Timon lepidus in France

Table 2. Input variables for the six models. See Table 1 for descriptions of variables.

Code Model description

M1 six selected bioclimatic variables

M2 eight random bioclimatic variables (blind test)

M3 12 climatic variables from Météo France

M4 M1 variables + solar radiation + PET + aridity

M5 M1 variables + altitude, slope, aspect

M6 M1 variables + altitude, aspect + vegetation

The inputs were then optimized by testing the model with

other sets of variables, using a stepwise procedure to 1n-

clude or exclude variables one by one. Every iteration

was evaluated and outcomes associated with the chosen

parameters were recorded. This resulted in six models

based on different combinations of variables (Table 2).

While a comparison of bioclimatic data sources in

terms of usefulness for species distribution modelling is

beyond the scope of this study, a deliberate choice was

made to use various sources in the model; specifically,

bioclimatic data from entirely different inputs. This itera-

tive and multiple-source approach increases the chances

of obtaining a successful model by allowing the selection

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Source: The standard deviation of yearly NDVI relative to the mean NDVI

for the month of July was calculated from MODIS data captured on the first

week of July every year from 2000 to 2016.

Variables

Bio4, Bio9, Biol0, Biol4, Biol5, Biol6

Bio3, Bio4, Bio5, Bio8, Bio9, Biol3, Biol7, Biol9

TMO, TMN, TMX, TAM, TEH, TEE, PDH, PDE, PJH,

PEH, PEE, PRA

[Biol, Bio4, Bio9Bio15, Biol6] + srad + PET + Aridity

[Bio4, Bio9, Biol0, Biol4, Biol5, Biol6] + Alt, Slope,

Aspect

[Bio4, Bio9, Biol0, Biol4, Biol5, Biol16] + [AIt, Aspect]

+ NDVI

of the most accurate and relevant variables. It also helps

to validate overall model performance, by providing keys

for analyzing the suitability of the model in terms of bio-

climatic variables favorable to the species.

Model descriptions

Model 1: Variables selected from the CHELSA climate

dataset based on ecological assumptions. The first

modelling trial used a set of ecogeographical variables

identified based on expert knowledge of the biology

and behavior of the species. As the Ocellated Lizard is a

Mediterranean species that prefers long periods of warm

Vegetation cover variation from 2000 to 2016

Standard deviation of yearly NDVI relative to the

mean NDVI for the month of July

[JO - 0.01

[10.01 - 0.05

0.05 - 0.075

0.075 - 0.1

M0.1-0.2

- Occurence data from 2000 to 2016

Fig. 5. Change in vegetation cover from 2000 to 2016: standard deviation of the yearly NDVI values relative to the NDVI mean

value for the month of July.

Amphib. Reptile Conserv.

December 2019 | Volume 13 | Number 2 | e213

Table 3. Variable correlation values for Model 1.

Jorcin et al.

bio4 R bio9 R biol0_R biol4 R biol5_R biol6 R

bio4 R 1.00000000 -0.07440553 0.3081616 -0.2788248 0.1672821 -0.2605788

bio9_R -0.07440553 1.00000000 0.5115825 -0.4025089 0.3664137 -0.0604624

biolO_R 0.30816159 0.51158248 1.0000000 -0.6782349 0.6210210 -0.1046921

biol4 R -0.27882481 -0.40250891 -0.6782349 1.0000000 -0.6896546 0.2434781

biolS_R 0.16728213 0.36641372 0.6210210 -0.6896546 1.0000000 0.2159100

biol6_R -0.26057884 -0.06046240 -0.1046921 0.2434781 0.2159100 1.0000000

temperatures (for ecological reasons linked to its repro-

ductive cycle), this model considered the bioclimatic

variables that would best represent this need. Variables

with a recognized influence on a species’ ecology are

generally expected to lead to more accurate predictions

in SDMs (Austin 2002). This initial SDM incorporated

the following variables from the CHELSA database:

Bio4, Bio9, Biol0, Biol4, Biol5, and Biol6 (Table 3).

The Biol16 variable was selected as it represents precipi-

tation in the wettest quarter, so it would be a good in-

dicator of the aridity of the environment. This variable

differentiates regions according to precipitation patterns

by indicating rainfall occurring during the wettest period

of the year, thus allowing locations with lower rainfall

to be identified. After this a priori selection, the correla-

tions between variables were measured (Table 3). As the

variables were not correlated, this choice was validated.

Model 2: Non-correlated variables selected from the

overall CHELSA climate dataset. For the next model, the

correlations between the 19 variables obtained from the

original CHELSA database for all presence-data loca-

tions were calculated, removing one of each pair of high-

ly correlated variables (those with a correlation coeffi-

cient greater than 0.75) [Doorman 2012]. As 11 of the 19

input variables were correlated, only the remaining eight

were retained, with no consideration of any presumed

ecological significance. Thus, this trial was considered

a blind test, run on a statistical basis only rather than on

prior knowledge of the input variables. Problems of col-

linearity between variables were identified and dealt with

using variance inflation factors (VIF) with the R usdm

package (Naimi et al. 2014). A VIF was calculated for

each explanatory variable, and those with a VIF greater

than 10 were removed. The correlation coefficients of

the remaining variables ranged between -0.015 and 0.75

(Table 1 in Supplementary Materials).

Model 3: Non-correlated variables from the Meétéo

France climate dataset. As an alternative to the CHEL-

SA climate database, this model used a climate dataset

obtained from Météo France weather stations (Joly et al.

2010). Correlation tests showed that out of 14 variables

from the Météo France dataset, only two were highly

correlated, with a correlation coefficient greater than

0.75. Therefore, the 12 non-correlated variables were in-

cluded in the model, with each having a potential effect

on model performance.

Amphib. Reptile Conserv.

Model 4: The variables for Model I with the addition

of solar radiation, evapotranspiration, and aridity.

This model included the six climate variables selected

for Model | in addition to climatic parameters that are

potentially important to the ecology of the species. To

reflect discriminating factors related to the Mediterra-

nean climate, this model used mean annual solar radia-

tion (obtained from WorldClim), as well as the Global

Potential Evapotranspiration and Global Aridity Index

(obtained from CGIAR) [Fick and Hijmans 2017]. The

Global Aridity Index consists of mean annual precipita-

tion divided by mean annual potential evapotranspiration

(Zomer et al. 2008). An assessment of whether the per-

formance of the model increased with these additional

variables was made.

Model 5: The variables for Model I with the addition

of topographic variables. This model included the six

climate variables selected for Model 1 along with three

additional topographic variables: elevation, slope, and

aspect (orientation). Topographic parameters were ex-

pected to improve model performance (Humboldt and

Bonpland 1805).

Model 6: The variables for Model I with the addition

of selected topographic variables and a vegetation

variable. The final model included the six climate vari-

ables selected for Model 1 along with two additional

topographic variables (elevation and aspect) as well as

NDVI. The two topographic parameters were retained

because of the gain in performance obtained by add-

ing them to the bioclimatic parameters. The addition of

the NDVI helped to account for vegetation cover as a

contributing variable in the model, as this is a valuable

parameter in identifying the natural habitat of the spe-

cies. Assuming the distribution of the Ocellated Lizard

is linked to this species’ preferences in terms of land

cover, vegetation density is expected to help differenti-

ate areas of occurrence from areas of absence (Wilson

etal 2013),

SDM methods

To maximize SDM accuracy, all six models were run

with eight statistical algorithms (Bucklin et al. 2014),

regression-based machine learning, and classification

methods. With each algorithm resulting in different pre-

December 2019 | Volume 13 | Number 2 | e213

Distribution of Timon lepidus in France

Table 4. Algorithms used in the species distribution modelling.

Code Description

BRT Boosted Regression Trees

CART Seek rae Regression Trees for

GAM Generalized Additive Model

GLM Generalized Linear Model

MARS Multivariate Adaptive Regression Spline

MAXLIKE Maximum Likelihood

RF Random Forests

SVM Support Vector Machine

dictions, the objective was to identify the method that

achieved the best accuracy (Elith et al. 2006). Testing

eight algorithms also allowed the evaluation of the over-

all modelling approach (Table 4). As well as analyzing

discrepancies between models in terms of performance,

model congruence was examined to consolidate the

conceptual approach (Li and Wang 2012). The model-

ling methods used were Generalized Linear Modelling

(GLM, Guisan and Zimmerman 2000), Generalized Ad-

ditive Modelling (GAM, Guisan and Zimmerman 2000),

Multivariate Adaptive Regression Spline (MARS, Elith

and Leathwick 2007), Maximum Likelihood (MAX-

LIKE), Classification and Regression Trees for Machine

Learning (CART, Breiman et al. 1984), Boosted Regres-

sion Trees (BRT, Elith et al. 2008), Support Vector Ma-

chine (SVM, Drake et al. 2006), and Random Forests

(RF, Breiman 2001).

The presence-absence models were used with the ob-

jective of predicting the presence probability of the Ocel-

lated Lizard and mapping its distribution accordingly.

Lacking absence data, pseudo-absences were used to

run the models. Pseudo-absence data was generated with

the R sdm package (Naimi and Araujo 2016), which has

the advantage of providing a pseudo-absence selection

process calibrated to SDM performance by considering

presence data. Other studies have found that randomly

selected pseudo-absences yield the most reliable models

(Barbet-Massin et al. 2012). The models were fit by as-

signing a number of pseudo-absences weighted to pres-

ences (Barbet-Massin et al. 2012), with an equal number

of presences and absences.

Each type of SDM methodology has particular strengths

and limitations in the way it accommodates the responses

to predictors, as well as how it deals with missing observa-

tions. For example, linear regression fits linear functions

relating a response variable to one or more predictor vari-

ables, where this relationship can be approximated by a

straight line (Ferrier et al. 2002), whereas machine learn-

ing offers more complex classification algorithms that

accommodate non-linear variable interactions (Salas et

al. 2017). All algorithms were tested with replicated sub-

sampling of 20% of the occurrence dataset.

Amphib. Reptile Conserv.

Model evaluation methods

The contribution of SDMs in understanding the

geographical distribution and abundance of a species

depends on the level of reliability offered by the model

(Barry and Elith 2006). The prediction accuracy must

be assessed to determine the model’s suitability (Liu

et al. 2009). Models can be judged on their capacity to

discriminate presence from absence, which is measured

by the number of false positive and false negative

predictions. Several statistical indicators can be used as

metrics to evaluate model performance (Fielding and

Bell 1997). To assess the results here, the area under the

Receiver Operating Characteristic (ROC) curve (AUC)

value was used, as well as the correlation coefficient

(COR) and the True Skill Statistics (TSS) value (Bradley

1997). The AUC value provides a single measure of

model performance, showing the model’s ability to

rank a randomly chosen presence observation higher

than a randomly chosen absence observation (Liu et

al. 2009). These values can range between 0 and 1; and

models producing AUC values of 0.75 are regarded as

reliable, 0.8 as good, and 0.9 to 1 as having excellent

discriminating ability (Franklin 2009). The TSS is

presented as an improved measure of model accuracy,

defining the average of the net prediction success rates

for presence sites and for absence sites (Allouche et al.

2006). The COR value allows another performance index

comparison between models, and helps to validate the

results obtained by the AUC and TSS methods (Elith et

al. 2006). This study also ran null models, which make

predictions in the absence of a particular ecological

mechanism (Harvey et al. 2003), to assess the random

probability hypothesis. The AUC values from the null

models ranged from 0.51 to 0.53, thus corresponding to

what would be expected by chance (Raes and ter Steege

2007). The distribution generated with a null model

significantly differed from the other modelling results,

with a predicted presence spread smoothly over most of

the study area (Fig. 2 in Supplementary Materials).

As well as evaluating model performance with

statistical indicators, the results were checked against

the validation dataset (Anderson et al. 2003), consisting

of the 25% of available presence data set aside for

validation. The data distribution showed that 76%

of the observation records within the presence range

correspond to a threshold value of 0.9 to 1, and 86% were

above a threshold of 0.8 (Table 5). Of the 690 occurrence

locations set aside to check the results, 92% fell within

the area predicted by the modelled map, in line with a

presence probability threshold of 0.70.

Predictive maps generated by SDMs provide the oc-

currence probability of the species on a 0 to 1 scale.

Threshold determination is a key step in transforming 1n-

dices of suitability to binary predictions of species pres-

ence or absence (Nenzen and Araujo 2011). Threshold

definition can be subjective or objective (Manel et al.

December 2019 | Volume 13 | Number 2 | e213

Jorcin et al.

Table 5. Correspondence between observations and presence

probability for the validation dataset. Values above a threshold

of 0.7 are shown in bold.

Probability threshold Sonate -

0.9 to 1 2,219 80.4

0.8 to 0.9 266 9.7

0.7 to 0.8 114 4.1

0.6 to 0.7 80 20

0.5 to 0.6 43 ne

0.4 to 0.5 13 a

0 to 0.4 22 ue

1999), and in many methods, the point at which sensitiv-

ity (true positive rate) and specificity (true negative rate)

are equal can be chosen to determine the threshold. In

this case, a value of 0.7 was chosen.

Results

All SDMs for the differing sets of variables performed

well, demonstrating high predictive power (Table 6). The

overall mean AUC value was 0.86, the mean TSS was

0.67, and the mean COR was 0.61. These results con-

firmed the hypothesis that SDMs based on bioclimatic

variables could provide valuable results for the Ocel-

lated Lizard. Moreover, the agreement between results

validates the overall modelling methodology, including

the quality of the sampling dataset and the geographical

extent of the study. Overall, the initial trial made with

Model 1 showed that it was a satisfactory model, with

an AUC value of 0.91, a TSS value of 0.72, and a COR

value of 0.78. The subsequent iterative trials carried out

with different combinations of variables further increased

model accuracy, validating the most useful variables and

helping to rank their contributions.

A comparison of the overall results led to the selection

of Model 6 generated with the RF method as the most

accurate model. The predictors of this model, based on

selected bioclimatic variables with additional topograph-

ic and vegetation parameters, achieved the best perfor-

mance using virtually all modelling methods, with maxi-

mum performance obtained from the RF method. This

combination of variables and modelling method resulted

inan AUC value of 0.98, a TSS value of 0.85, and a COR

value of 0.88. An analysis of variance (ANOVA) was

performed to confirm the validity of the chosen combina-

tion of variables. ANOVA results on Model 6 showed the

lowest p-values for all variables, with a minimum value

of 2.74e-14 and a maximum value of 0.020393 (Table 7).

In contrast, models 1 to 5 each included a variable with

a p-value > 0.05. As a study by Wood et al. (2016) men-

tioned that the Akaike information criterion (AIC) works

reasonably well for model selection, the AIC between

models were also compared here, and this comparison

showed that Model 6 had the lowest AIC values (Table 2

in Supplementary Materials).

Contribution of Variables

The contributions of each variable in Model 6 were

ranked, identifying four critical parameters, as well as

one secondary factor (Table 8). The four main factors

that most influenced model performance were (according

to their relative importance): precipitation in the driest

month, temperature seasonality, mean temperature 1n the

driest quarter, and NDVI. Precipitation seasonality was

Table 6. Comparison of model results based on different modelling methods and assessments of model performance. The values in

bold, for Model 6 and RF, indicate the results with the highest accuracy.

x [os [aie

P06

a A

ws [oso [oss | os | 0s [oor [oi | os | 07 | o7 | oss [057 | 05

[ms [082 [oss | oa [ 085 [06s | oe | oor | oo | o6 | oss | 057 [ost

[auc [cor [1s [auc | cor [ 18s | auc | cor | 1s | auc [ cor] 155

Twi [oss [or [oss [082 | os0 | oss | oor | o7 | om | oss [on | 06s

To [oss [068 [oss | oss [oo | oso | om | os | o7s | os [075 [om

Twa_[ os [or | 046 | om [os4 | 032 | 09 | 07s | o73 | ome | 075 [on

Twa | oss | 069 | 06s | 079 [oss | oas | oo | 079 | om | ome | om | om

Tos [or | oar | oas | oss | 049 | oss | oa | o76 | oa | om | om

| Me | 088 | o65 | 059 | 082 | 056 | 048 | oss | 087 | 08s | 09 | 07 | 067 |

December 2019 | Volume 13 | Number 2 | e213

Amphib. Reptile Conserv. 285

Distribution of Timon lepidus in France

A 0 50 100km

N mater

A? 50 100 km

A 0 50 100km

Presence

probability

i o-o

[_]0-0.05

[_] 0.05 - 0.1

[J 0.1 - 0.15

[0.15 - 0.2

[9 0.2 - 0.25

[) 0.25 - 0.3

fl 0.3 - 0.35

[i 0.35 - 0.4

I) 0.4 - 0.45

i 0.45-0.5

Ml 0.5 - 0.55

Ml 0.55-0.6

MM 0.6 -0.65

Hl 0.65 - 0.7

i 0.7 - 0.75

MW 0.75-08

MH o8-0.85

MM 085-09

MM 0.9-0.95

MM 0.95-1

MODEL 4

A 0 50 100 km

—

Fig. 6. Predictive modelling maps showing presence probability of the Ocellated Lizard (Timon lepidus) in the study area.

also a contributing variable, but only to a minor extent.

Climatic factors associated with dry and warm weather

conditions seem to play a determining role in the spatial

distribution of the Ocellated Lizard, with precipitation in

the driest month being the primary contributing factor.

The contributions of these variables can be interpreted as

a reflection of the species’ ecological needs, especially as

related to its reproductive cycle.

Predictive Habitat Suitability Maps

The predictive maps (Fig. 6) show very slight differences

between the six models. Oléron Island, where the most

northerly currently known population of this species is

found, is included in all models, but with different prob-

ability ranges. Its presence is predicted on the whole is-

land in models 1, 2, 4, and 5, but only on part of the island

in models 3 and 6. All models show a clear link between

the ‘Mediterranean population’ and the ‘Lot population’

(in a region lying northwest of the Mediterranean), with

minor variations in the continuity of the species distri-

bution between these two ‘populations.’ In the different

maps, the penetration of the species into the Rhéne Val-

ley appears more or less extended, and the fragmentation

of the “Lot population’ is more or less pronounced. Apart

from these details, the maps based on the six models are

extremely consistent.

Amphib. Reptile Conserv.

On the map generated by Model 6 (Fig. 7), most of

the locations historically occupied by the species (1.e.,

for which evidence exists of its disappearance) are along

the Atlantic coast (in the Nouvelle-Aquitaine region), the

area where the distribution of the species 1s the most lim-

ited and fragmented (Fig. 8). The disappearances of the

three Mediterranean populations correspond to very spe-

cific cases: two islands (Ratonneau and Porquerolles is-

lands), where the species is likely to have disappeared as

a result of the introduction of predators (Cheylan 2016),

and a population in the Rhdéne delta in the Camargue,

where the decline of rabbits has transformed the envi-

ronment, leading to the disappearance of the Ocellated

Lizard in this area (Doré et al. 2015).

Identification of Knowledge Gaps

The potential distribution predicted by Model 6 indi-

cates that knowledge gaps regarding the observed pres-

ence of this species are considerable, not only in the

Mediterranean distribution range, but also in its pe-

riphery (Fig. 9). Allowing a buffer zone with a 5-km

radius around each observation location, only 74% of

the area of predicted presence (based on Model 6 with

a presence probability threshold of 0.70) is confirmed

by actual observations; observation data is lacking for

26% of the area. In the core of the distribution range,

December 2019 | Volume 13 | Number 2 | e213

Jorcin et al.

0 2550 100km

7 elicomeh

e Presence data collected 2000-2016

® Predicted distribution of Ocellated Lizard

Bi, WG, RAL NPS, RAIA, depots KIM Kaede NL. Cecnance Sucany, Ean depen

} Sapereparemn, iB ipe aD 00 a aved Tee GIS Lege Con

Fig. 7. The predicted distribution map generated by Model 6 (in pink) with specific locations of presence data (from observations)

from the dataset (black dots).

information is missing in several areas of the regions

of Provence and in a few areas of Languedoc-Roussil-

lon. This is particularly the case for several noteworthy

zones.

¢ Var department (e.g., around Fayence, Draguig-

nan, Bargemon, Seillons-source-d’Argens, and

Saint-Maximin-la-Sainte-Baume)

¢ Southeast of Alpes-de-Haute-Provence (e.g., the

Valensole plateau, the Asse-Puimichel valley,

and the lower Bléone valley)

¢ Southeast and northwest of the Vaucluse depart-

ment (e.g., Pertuis, La Tour-d’Aigues, Carpen-

tras, Uchaux, Sainte-Cécile-les-Vignes, and

Valréas)

¢ Dréme department (Valence plain, Monteéli-

mar plain, Tricastin, and Baronnies Proven-

cales)

¢ Gard department (e.g., Saint-Quentin-la-Poterie,

Bagnols-sur-Ceze, Ales, and La-Grand-Combe)

¢ Southeast of Ardeche (e.g., Saint-Thome, Vil-

leuneuve-de-Berg, and Saint-Marcel-d’ Ardeéche)

¢ South and far northwest of the Lozere (e.g.,

Aysseries, Faveyrolles, La-Bastide-Solages, Sé-

brassac, Decazeville, and Claunhac)

In western Languedoc-Roussillon, it would seem rel-

evant to look for the Ocellated Lizard in several zones

of the Aude department (e.g., Carcassonne, Labécede-

Lauragais, Rouffiac-des-Corbieres, and Palairac),

and in several areas of the Pyrénées-Orientales (e.g.,

Saint-Paul-de-Fenouillet, and La Trinité) and the

Amphib. Reptile Conserv.

Herault (e.g., Pézenas and Aigues-Vives). Outside of

the Mediterranean region, areas where this species

would merit further survey efforts are more numerous,

notably in the Aveyron, Tarn, Haute-Garonne, Tarn-et-

Garonne, Lot, and in Dordogne, where the potential

distribution area is very fragmented and the natural

habitats small (Berroneau 2012; Pottier et al. 2017).

To address this, surveys could be carried out in several

areas of the Tarn and the Lot (e.g., Mazamet, Roque-

courbe, Larroque, Lacapelle-Marival, Prayssac, Gour-

don, and Martel), in southwest Correze (e.g., Tulle,

Taurisson, and Saint-Aulaire), in southeast Dordogne

(e.g., Carsac-Aillac, Hautefort, and Rouffignac-Saint-

Cernin-de-Reilhac), and in eastern Tarn-et-Garonne

(e.g., Caylus). All of these areas possess a high pres-

ence probability of this species according to the model

results, so it 1s likely that inadequate surveying ex-

Table 7. ANOVA produced by the Generalized Additive Model

for Model 6. Approximate significance of smooth terms.

Edf Ref.df Chi.sq p-value

s(bio4_R) 7.928 8.263 84.87 2.74e-14

s(bio9_R) 7.445 7353 38.47 5.88e-06

s(biol10_R) 6.469 6.779 42.75 7.37e-07

s(biol4_R) 7.608 8.248 18.61 0.020393

s(biol5_R) 6.071 7.297 18.72 0.008308

s(biol16_R) 7.944 8.688 18.79 0.018410

s(alt250_R) 6.279 6.819 34.27 9.87e-06

s(orient250_R) 3.392 4.241 20.47. ~~ 0.000507

s(median MO- 2.734 3.462 40.17 3.36¢e-08

DIS2)

December 2019 | Volume 13 | Number 2 | e213

Distribution of Timon lepidus in France

Pays _de la Loire

<— La Rochelle

Atianticfes, ff ~~ & oe epee :

coast ' Ce.

Nouvelle-Aquitaine

F ¥ ee

yCancnms

Guara

Hue

Centre-Val de Loire Bourgogne-Franche*

ie Montlugon

tong i -

Catalanes a ta

Sources: Esm! HERE, DeLorme, Intermap, increment P Corp., GEB

METI, Esri

Comtéx,

Lyon PRR af

Bourgoin iy ddacail &

falieu at all

ibeéry

Auvergne-Rhéne-Alpes

tee

A - : bie ade

U7 Camargues .

' Ratonneau<®

island

Porquerolles

i island

e@ Disappeared populations

[| Regions boundaries

NN) Predicted distribution

Fig. 8. Location of populations that have disappeared in relation to the predicted presence map generated by Model 6 (blue dots).

plains the current absence of data in these locations.

Contribution of Protected Areas to the Conservation

of the Species

A comparison of the potential niche (Model 6 with a

presence probability of > 0.70) and the main national

protected areas allowed an assessment of the contribu-

tion of the current network of natural reserves to the con-

servation of the species (Table 9).

The map in Fig. 10 shows the protected areas that

contribute most to the conservation of the species are the

regional nature parks (representing 22% of its predicted

niche) and the two Natura 2000 zones (15% and 16%).

The other types of protected areas contribute to a much

more limited extent, covering between only 0.1% and 2%

of the potential distribution area of 35,805 km?. As these

protected areas often overlap (regional nature parks of-

ten include nature reserves, and are generally also part of

the Natura 2000 network), it 1s difficult to calculate the

surface areas to evaluate the total contribution of all the

protected sites. It should be noted that the protected areas

with the strongest regulatory protection (national parks,

national and regional nature reserves, and National For-

est Agency ecological reserves) cover less than 1% of the

potential niche of this species.

Amphib. Reptile Conserv.

Discussion

Model Performance

Nine variables, out of the selection of 28 climate vari-

ables, three topographic variables, and three vegetation

variables, were found through iteration to contribute

most effectively to the quality of the models. The tests

of six models with eight statistical algorithms led to very

sound results, confirming the performance of the models.

The consistency between the models and the statistical

soundness of the modelling are largely due to the good

spatial coverage of the source data, its geographical pre-

cision, and the size of the dataset (Araujo and Guisan

2006).

Given the exhaustive nature of the occurrence data, we

feel confident that the predictive model of the ecological

niche gives a fairly good picture of the potential distribu-

tion of the species, and can help to map its actual current

distribution. Other studies have demonstrated that SDMs

can accurately determine the natural distribution of a spe-

cies, contributing to the more complete knowledge of its

current range (Elith and Leathwick 2009).

In this study, the RF method resulted in the most accu-

rate SDMs, performing well with all the sets of variables.

This method uses decision trees based on random group-

ing of the covariates, modelling both the interactions be-

December 2019 | Volume 13 | Number 2 | e213

Jorcin et al.

Table 8. Variable importance index generated by the best SDM, sorted by rank of importance.

Variable Description

biol4 Precipitation in the driest month

bio4 Temperature seasonality (std* 100)

bio9 Mean temperature in the driest quarter

ndvi Normalized Vegetation Index

biol5 Precipitation seasonality (coefficient of variation)

biol0 Mean temperature in the warmest quarter

biol6 Precipitation in the wettest quarter

altitude Elevation

aspect Slope orientation

tween the variables and their nonlinear relationships, and

it uses bootstrapping to fit individual trees (Salas et al.

2017). Cutler (2007) demonstrated that the advantages

of RF include very high classification accuracy, ability

to model complex interactions between predictor vari-

ables, and an algorithm for imputing missing values.

This gives RF the flexibility to perform several types of

statistical data analysis, including regression, classifica-

tion, survival analysis, and unsupervised learning (Cutler

2007). Rangel and Loyola (2012) also demonstrated that

machine learning methods such as RF have high statisti-

cal precision and predictive power for determining the

species distribution of well-known populations.

While the models here showed high accuracy, certain

improvements could be made by integrating additional

4 Soret Ga

a wit

CHARENTE-MARITIME 'CHARENTE

Angoulém:

GIRONDE"*

LANDES... 4.

Marsan

PYRENEES-ATLANTIQUES E

mile & HAUTE-GARONNE

Tarbes

/ woe HAUTES-PYRENEES = :

Va N A, era UY ARIEGE

Pyeaies ?

0 25.50 100km

hor

"a Lf aes ea ra N. Sierra

METL Een chin a (Hong Kong), swisstopo

Cor_Test AUC test Rank

0.1794 0.0330 1

0.1172 0.0216 2

0.1012 0.0168 3

0.0801 0.0152 4

0.0335 0.0060 6

0.0394 0.0018 7

0.0575 0.0012 8

0.0219 0.0004 9

datasets not included in this study, such as soil maps, the

distribution of tree species, or detailed vegetation maps

of France (Leguédois et al. 2011). While land cover

parameters could contribute to identifying habitat suit-

ability, their integration in SDMs remains a challenge

in terms of accuracy and validity (e.g., in terms of how

current they are). The challenges of incorporating land

cover data are due to the fact that they are categorical

variables with limitations imposed by spatial resolution,

date of production, and thematic classification. Studies

have shown that continuous remotely sensed predictor

variables offer many advantages over categorical vari-

ables and can be used effectively in species distribution

modelling (Wilson et al. 2013). Models based on biocli-

matic variables have proven their efficiency in numerous

Ferrand

PUY-DE-DOME

SAVOIE

Nato ralde

= HAUTE-LOIRE

PNR fi i *

ARDECHE ~

- LOZERE

eee : AS

Piece

[ __] Department borders

i Predicted distribution without any

observations within 5 km

Fe Re Narr oa aera

al Predicted distribution with

observations within 5 km

Fig. 9. Areas within the potential niche of the Ocellated Lizard (in orange) for which there are no observation data (in red), based

on Model 6 with its presence probability threshold of 0.70 combined with presence data from observations including a buffer zone

with a 5-km radius.

Amphib. Reptile Conserv. 289 December 2019 | Volume 13 | Number 2 | e213

Distribution of Timon lepidus in France

Table 9. The contributions of the different types of protected areas to the conservation of the Ocellated Lizard in France. The surface

area favorable to the presence of the species (“potential niche” in header of second column) is given in km? according to Model

M6. The percentage of the protected area relative to the total surface area of the predicted distribution range is shown on the third

column.

Surface area under protection within

Percentage of the potential

ype OUpnovectvares the total potential niche (km?) niche*

Regional Nature Park 8,121 22.68

Special Area of Conservation (Natura 2000) 5,927 16.55

Special Protection Areas (Natura 2000) 52985 15.60

National Park (park peripheral zone) 792 221

National Nature Reserve 170 0.47

National Park (park core area) 92 0.26

Ecological Reserve (National Forest Agency) 43 0.12

Regional Nature Reserve 20 0.06

* Total surface area predicted with a probability threshold of 0.7 = 35,805 km’.

studies, while one study has shown that the addition of

land cover variables to pure bioclimatic models does not

necessarily improve the predictive accuracy of the result-

ing SDM (Thuiller et al. 2004).

Consistency between Potential and Realized Distribu-

tions

The results obtained here showed strong agreement be-

tween the predictive models and the observed distribu-

tion of the Ocellated Lizard on a macro-geographic scale,

giving rise to several conclusions. The first is that climat-

ic predictors prevail over all the other predictors for this

species. The current boundaries of the distribution range

Parc’

ional

Pamplona/lruna re

National des

Saach 400 km Renae! ‘

ae Andorra = > Me

of the Ocellated Lizard in France are essentially defined

by climatic factors, which aligns with many studies on

reptiles (Guisan and Hofer 2003; Santos et al. 2009; Bri-

to et al. 2011). This suggests an ancient presence of the

Species in France, given the barriers to dispersion such as

rivers and mountains in its territory, and the time neces-

sary to colonize the entire potential bioclimatic niche. The

fragmentation of the populations at the edges of the dis-

tribution, as well as the historical information regarding

the loss of populations (Cheylan and Grillet 2005; Doré

et al. 2015), support this idea and suggest the existence

in the past of a larger and, above all, a less fragmented

range. Unfortunately, zooarchaeological information on

this subject is limited. The species is known to have been

Saint

‘Ss

tc aE aco

La Predicted distribution within

protected areas

HERE, DeLorme, Interma

METI. Esti China (Hong Kong), naa Mapi P re d icted d ist ri b utio n

outside of protected areas

Figure 10: Contribution of protected nature areas to the conservation of Timon lepidus. Predicted presence within protected areas

(dark purple) and predicted distribution range (light purple).

Amphib. Reptile Conserv.

290

December 2019 | Volume 13 | Number 2 | e213

Jorcin et al.

present in France in the Middle Pleistocene (~700,000

to 150,000 yr ago), from remains in the Lazaret cave in

Nice (Bailon 2012), and remains from the Holocene have

also been found (Mateo 2011). However, the lack of fos-

sil remains from the last interglacial optimum (between

125,000 and 11,000 yr ago) does not definitively prove

the retreat of the species to the Iberian Peninsula dur-

ing this period. The presence of isolated populations on

the northern edges of the current distribution, as well as

its presence in Liguria, would have required overcoming

major obstacles (the Rhéne, Var, and La Roya rivers),

which points to an ancient occupation of the territory.

Given these factors, the hypothesis that the species re-

mained during the interglacial period seems possible, at

least in the far south of France.

The strong concurrence between the models and the

observed distribution also indicates that the process of

decline in this species is moderate, as all the areas favor-

able to the species are still occupied, apart from a few

exceptions. Even at a lower spatial resolution, the bound-

aries of the distribution range are primarily due to cli-

matic factors, and only secondarily to ecological factors

(i.e., presence of favorable habitat). This is particularly

true at the edges of its distribution range in the valleys

that open onto the Mediterranean coast (1.e., those of the

Aude, Rhéne, Durance, and Var), where the extent of the

penetration of the species coincides with the boundary

of the Mediterranean climate and vegetation, as there

are no physical obstacles preventing a deeper advance in

these valleys (Deso et al. 2011, 2015). Notably, the mod-

el clearly differentiates areas favorable to the Ocellated

Lizard in zones of rugged terrain. This is particularly the

case in the region of the Causses (in the southern part of

Aveyron), which is characterized by limestone plateaus

that would potentially be favorable to the species but

where it 1s not present, and by deep gorges (the valleys of

the Tarn and the Jonte) where the species has long been

observed. Rather surprisingly, the model distinguished

between these two zones (plateaus and gorges) despite

any notable climatic difference between them.

On the other hand, several areas not predicted by the

model have the proven presence of the species (e.g.,

the foothills of the Pyrénées in Ari¢ge, the mountain-

ous zones of Ardéche, and northern Dordogne). This is

likely explained by the resolution of the model, which is

ill-adapted to predicting very small areas within a land-

scape and climate matrix that is generally unfavorable to

the species. These known populations live in very small

micro-habitats (a few dozen ha at most) with unique

botanical characteristics distinct from the surrounding

landscapes. An analysis that takes into account a finer

landscape scale, particularly in terms of vegetation,

would produce a model with a closer fit. Equally, sub-

strate characteristics, which were not taken into account

in the model, play an important role in the presence of

the Ocellated Lizard when the climatic environment is

unfavorable. In this case, it seeks out terrain that is rather

Amphib. Reptile Conserv.

steep, rocky, or well drained to avoid environments that

are too wet.

Which Variables Best Explain the Distribution of the

Species?

At the macro-geographic scale, the variables that best ex-

plain the distribution of the species are related to climate

and, to a lesser extent, vegetation and topography. This

indicates the primacy, over all other variables, of a hot

and dry summer, as well as a strong seasonal contrast;

two key characteristics of the Mediterranean climate

(Blondel et al. 2010). The importance of temperature and

aridity in the summer Is certainly due to the reproduction

requirements of this species. In France, female Ocellated

Lizards are known to typically lay their eggs at the end

of May or the beginning of June (Cheylan and Grillet

2004), and the eggs hatch the third week of September

or the first week of October (Bischoff et al. 1984; Doré

et al. 2015). In Provence, this corresponds to an incuba-

tion period of about 100 days (Cheylan and Grillet 2004).

Hence, the entire summer period is used for reproduc-

tion. The late hatching period requires mild temperatures

at the end of summer and beginning of autumn, allow-

ing the hatchlings to feed before the hibernation period,

which begins around 15 November in most of the French

regions where this species is present (Doré et al. 2015).

At the local scale, the presence of the species 1s pri-

marily influenced by the aridity of the habitat; the Ocel-

lated Lizard prefers a rocky or well-drained substrate

that is well exposed to the sun. Dense vegetation cover

is very unfavorable for this species, as shown in a study

by Santos and Cheylan (2013) in Provence. In the future,

gaining a better understanding of the importance of each

of these habitat variables would be useful, drawing upon

the resources available on the subject.

Why is this Species Retreating at the Edges of its

Distribution Range, in Contrast to Climatic Expecta-

tions?

The proven extinction of several Ocellated Lizard popu-

lations over the last 150 years, mainly on the northern

border of its distribution range (Cheylan and Grillet

2005; Grillet et al. 2006) runs counter to what might be

expected with the warming of the climate, the effects

of which have been clearly demonstrated on Mediterra-

nean reptiles in the south of France (Prodon et al. 2017).

Given its high thermal requirements, this species should

in fact benefit from climatic warming, particularly at the

northern edge of its distribution. However, the opposite

is observed, which suggests the predominance of local

over global factors. Studies carried out to investigate this

issue have shown that several local factors explain the

decline (or even the disappearance) of local populations

of this species. Those factors include the introduction

of predators in the case of island populations (Cheylan

December 2019 | Volume 13 | Number 2 | e213

Distribution of Timon lepidus in France

2016), the disappearance of the European rabbit and the

resulting changes to the landscape (Grillet et al. 2010),

the impact of pest control on entomofauna prey (Doré

et al. 2015), and the abandonment of agricultural land

and the resulting progression of woodland (Grillet et al.

2006; Pottier et al. 2017). Thus, the expected effects on

reptiles of the changes caused by warming in Europe

(Araujo et al. 2006) are not borne out in the case of the

Ocellated Lizard.

Which Zones should be Surveyed in the Future to Im-

prove Our Knowledge of this Species’ Distribution?

The recent discovery of a population in Vendée (Cédric

Baudran, pers. comm. 2018), beyond the known bound-

ary on the Atlantic coast, shows that new populations

remain to be discovered, especially at the edges of the

distribution range. A priority would be to seek confirma-

tion of the true disappearance of the species in selected

sites where it was known in the past, based on the pres-

ence predictions generated by the SDMs. Secondly, an

attempt to confirm the existence of connections between

population clusters that are considered to be separated

would be interesting. This would be particularly useful

for populations located in the mountainous zones of the

Alpes-Maritimes (Deso et al. 2015), in the upper Durance

valley (Deso et al. 2011), and in the Rhdéne valley (Doré

et al. 2015), as well as the fragmented populations in the

Lozere, Aveyron, Tarn, Tarn-et-Garonne, Lot, Dordogne,

Correze, and Cantal (Geniez and Cheylan 2012; Pottier

et al. 2017). The coastal populations of the Atlantic cur-

rently seem rather well-defined (Berroneau 2012); how-

ever, this does not exclude the possibility of discovering

new populations there.

What Conservation Strategy should be Adopted to

Protect this Species?

The predictive distribution models generated in this

study provide interesting leads for defining a conserva-

tion strategy for this species. First, the current network of

protected areas in France can be considered to rather sat-

isfactorily cover the distribution range of the Ocellated

Lizard and its different population clusters. However, a

deeper analysis reveals that only a very small proportion

of the area potentially favorable to the species benefits

from strong protection regulations. The areas of land

with the strictest protection (national and regional nature

reserves, National Forest Agency ecological reserves,

and national parks) only represent 1.2% of the potential

niche of this species in France. In terms of national parks,

the Cévennes National Park clearly bears the most re-

sponsibility in terms of the conservation of this species,

followed at some distance by the Calanques National

Park (respectively, 40 and 95 km? of favorable habitats

for the species). There are 15 national nature reserves

with the presence of the species, and in this category of

Amphib. Reptile Conserv.

protected area, the reserves of Coussouls de Crau and the

Maures plain have the largest known populations (re-

spectively, 74 km? and 52 km? of favorable habitat). Of

National Forest Agency ecological reserves, 16 include

land where the lizard 1s found, with the largest being the

Maures reserve (18 km? of favorable habitat) and the Pe-

tit Luberon reserve (16 km? of favorable habitat). Some

240 Natura 2000 sites have conditions that are potentially

favorable to the species; 21 protect areas of land that con-

tribute to the conservation of the species of more than 90

km, for a total contribution of about 3,430 km? for this

category of protected area.

Given the rather dense network of protected areas,

both in terms of spatial extent and altitudinal range, a

strategy based on anticipating climate change (Salas et al.

2017) is not necessarily the best choice. As stated above,

this species is in decline at the northern edge of its dis-

tribution, which runs counter to the expected effects of a

warming climate (which is predicted for the region in the

future). Moreover, the refuge habitat for this species (and

where it originated) is located in the southern half of the

Iberian Peninsula (Miraldo et al. 2011), so the effects of

climatic warming are unlikely to be harmful to the spe-

cies at the northern edge of its distribution. In support

of this hypothesis, it has been argued that the increase

in wildfires due to climatic warming will significantly

increase the density of Ocellated Lizard populations in

the Mediterranean region, by transforming woodland

into open landscapes (Santos and Cheylan 2013). A more

important consideration than climatic warming for the

conservation of the species is that its spread relies on the

existence (or not) of favorable environments, and its de-

mographic capacity to colonize new territories. Unfor-

tunately, studies of the isolated populations at the edges

of the distribution range (Grillet et al. 2006; Deso et al.

2015; Pottier et al. 2017) show that these two parameters

are rarely present, and that these populations are, in the

more or less long term, undergoing a process of extinc-

tion (Salvidio et al. 2004; Cheylan and Grillet 2005).

From a strategic point of view, therefore, the core of

the distribution range should be prioritized for conser-

vation efforts in the long term, without neglecting cer-

tain peripheral populations in the shorter term (e.g., the

populations in the valleys of the Durance, Rhdéne, and

Var rivers, and the sandy habitats of the Atlantic coast).

Our SDM-generated maps indicate that the isolated pop-

ulations of the Atlantic coast, as well as the population

clusters west of the Massif Central (in the departments

of Aveyron, Tarn, Tarn-et-Garonne, Lot, and Dordogne),

offer climatic and topographical conditions that are very

favorable to this species. These populations, while isolat-

ed from the Mediterranean population, should be given

careful attention. They may even harbor specific genetic

compositions that warrant further consideration.

The strong dependence of the Ocellated Lizard on

the European rabbit in soft soils (Grillet et al. 2010) also

suggests the value of taking concerted conservation mea-

December 2019 | Volume 13 | Number 2 | e213

Jorcin et al.

sures that equally protect this mammal. As the demo-

graphic trends of rabbit populations in the Mediterranean

region are very negative (Ward 2005; Delibes-Mateos

et al. 2008; Poitevin et al. 2010), this could result in a

domino effect on Ocellated Lizard populations.

Conclusions

This analysis, carried out at the scale of France, reveals

that the distribution of the Ocellated Lizard is primarily

conditional on climatic factors, in particular the length of

the arid summer period. Further study at a smaller scale

would help to provide a more detailed understanding of

the ecological preferences of this species. Such a study

could consider two finer, overlapping spatial scales: the

Mediterranean coast and the region around Montpellier.

Focusing on the Mediterranean coastal plains would al-

low climatic and topographic variables to be separated

out, at least partially, to better bring to light the roles of

factors linked to land use. Including the region of Mont-

pellier would more completely isolate climatic and topo-

graphic variables, allowing a focus on habitat variables

directly linked to the ecology of the species: soil type,

crop or natural vegetation type, level of urbanization, and

the presence and density of European rabbits.

Acknowledgements.—We would like to thank all the

observers and organizations that collected the data used

in this study: the nature conservation NGOs Cistude-Na-

ture, the League for the Protection of Birds in Provence-

Alpes-Céte d’Azur (LPO PACA) and the Dréme (LPO

Dréme), Nature en Occitanie, Méridionalis, and the pub-

lic platform for naturalist data, SILENE Provence-Alpes-

Cote dAzur. We would also like to thank Elise Bradbury

for reviewing the English text.

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Laurent Barthe is responsible for biodiversity at the NGO Nature en Occitanie in Toulouse, France. Lau-

rent has been president of the French Herpetological Society since 2017. Passionate about reptiles since

childhood, he is particularly interested in snakes and the European Pond Turtle. Two of his job roles are to

broaden knowledge about reptile and amphibian distribution and promote their conservation.

Matthieu Berroneau is a herpetologist at the NGO Cistude Nature in Bordeaux, France (http://www.

cistude.org), where he specializes in the herpetofauna of southwest France, with a particular emphasis on

conservation, education, and raising awareness. A lifelong interest in amphibians and reptiles also drives

Matthieu’s work as a professional photographer (http://www.matthieu-berroneau.fr), leading him to travel

the world to take pictures of a variety of species.

Marc Cheylan is a lecturer in Conservation Biology at the Ecole Pratique des Hautes Etudes (EPHE),

an institute within the PSL Research University (Paris Sciences et Lettres). Marc joined the EPHE’s bio-

geography and ecology research laboratory after a post as the associate curator at the Natural History

Museum of Aix-en-Provence. He currently works at the French National Centre for Scientific Research

(CNRS: http://www.cefe.cnrs.fr/fr/), Centre for Functional and Evolutionary Ecology (CEFE), based at the

University of Montpellier. Author of some 200 publications and four books in the fields of ecology, bio-

geography, and the conservation of amphibians and reptiles around the Mediterranean, Marc is a member

of several nature conservation organizations, including the International Union for the Conservation of

Nature (IUCN), and also sits on various scientific advisory bodies for natural parks and reserves.

Florian Doré is a naturalist who currently works at the NGO Deux-Sevres Nature Environnement in

Deux-Seévres, western France. Florian leads monitoring surveys and conservation studies on entomofauna

and herpetofauna, and has contributed to studies of the Ocellated Lizard since 2007. He co-authored the

National Action Plan for the Ocellated Lizard in France for the NGO OBIOS (Objectifs Biodiversités)

and co-authored a monograph on the species with Pierre Grillet and Marc Cheylan that was published by

Biotope Editions. Photo by Marc Cheylan.

Philippe Geniez is a research engineer in the Vertebrate Biogeography and Ecology lab at the Ecole

Pratique des Hautes Etudes (EPHE), an institute within the PSL Research University (Paris Sciences

et Lettres). The lab is part of the French National Centre for Scientific Research (CNRS: http://www.

cefe.cnrs.fr/fr/), Centre for Functional and Evolutionary Ecology (CEFE). Author of 236 publications,

Philippe is a specialist in Western Palearctic amphibians and reptiles. His research focuses on biological

systematics, phylogeny, ecology, and the distribution of plants and animals, particularly amphibians and

reptiles.

Pierre Grillet is a naturalist who, since 1995, has been studying the Ocellated Lizard at the edge of its

distribution, particularly on the island of Oléron, the last island population of the species in France. Pierre

has written several scientific articles and co-authored two books on the Ocellated Lizard. He regularly

organizes herpetological training for the French Agency for Biodiversity. Photo by Marc Cheylan.

Amphib. Reptile Conserv. 297 December 2019 | Volume 13 | Number 2 | e213

Distribution of Timon lepidus in France

Pierre Jorcin conducts research and leads projects on biodiversity conservation, with a focus on geospatial

modelling. Pierre’s areas of interest are species distribution, ecological niche modelling, and wildlife cor-

ridor mapping. He has been involved in sustainable development programmes in South Asia for 14 years.

Pierre is currently working on flora and fauna database management for ecological ranking and environ-

mental impact assessment studies in southern France.

Benjamin Kabouche is the director of the environmental NGO LPO PACA (Ligue pour la Protection

des Oiseaux, a member of BirdLife International) in the region of Provence-Alpes-Céte d’ Azur in France

(https://paca.lpo.fr/protection). One of Benjamin’s roles is to coordinate studies and nature conservation

programs. He has contributed to several naturalist publications on the subjects of terrestrial wildlife and

biogeography.

Alexandre Movia works for the environmental NGO LPO Dréme (Ligue pour la Protection des Oiseaux),

where he is an ecological corridor specialist. Alexandre acts as an advisor on herpetology to the French

department of the Dréme, and recently conducted a study on the distribution of the Ocellated Lizard in

this region.

Babak Naimi is a researcher at the University of Helsinki, Finland, with a research focus on modelling

species distribution and biodiversity under climate change and land use change scenarios. Babak is inter-

ested in developing a quantitative understanding of ecosystem dynamics (e.g., through remote sensing and

geoinformatics tools) and uncovering the complexities behind ecosystem behavior.

Gilles Pottier has been a professional field herpetologist for some 20 years, and currently works mainly

in the Pyrenees and the Massif Central for the NGO Nature en Occitanie. A member of the French Herpe-

tological Society, Gilles runs training events in herpetology in southwest France and has written numer-

ous papers and books about the local herpetofauna, including a work on the reptiles of the Pyrenees (Les

Reptiles des Pyrénées), published in 2016 by the French Natural History Museum. Photo by J.P. Vacher.

Jean-Marc Thirion is an ecologist and the director of the NGO OBIOS (Objectifs Biodiversités) in south-

west France. Jean-Marc leads conservation projects to protect natural areas and conducts population moni-

toring of amphibians and reptiles to promote conservation initiatives. He has participated in many natural-

ist surveys of flora and fauna.

Amphib. Reptile Conserv. 298 December 2019 | Volume 13 | Number 2 | e213

Official journal website:

amphibian-reptile-conservation.org

Amphibian & Reptile Conservation

13(2) [General Section]: 299-303 (e214).

First field report of Trimerodytes percarinatus (Boulenger,

1899) (Reptilia: Squamata: Natricidae) from India with notes

on its natural history

1*Ashok Kumar Mallik, 2Subhadeep Chowdhury, *Bharat Bhushan Bhatt, and *Ashok Captain

‘Centre for Ecological Sciences, Indian Institute of Science, Bangalore 560012, Karnataka, INDIA *Krishnachak, Dhurkhali, Howrah 711410, West

Bengal, INDIA *Department of Environment & Forest, P-Sector, Itanagar 791111, Arunachal Pradesh, INDIA *B-2, La Shanz Apartments, 3/1 Boat

Club Road, Pune 411001, Maharashtra, INDIA

Keywords. Arunachal Pradesh, distribution, Natricinae, new country record, Sinonatrix pericarnata, water snake

Citation: Mallik AK, Chowdhury S, Bhatt BB, Captain A. 2019. First field report of Trimerodytes percarinatus (Boulenger, 1899) (Reptilia: Squamata:

Natricidae) from India with notes on its natural history. Amphibian & Reptile Conservation 13(2) [General Section]: 299-303 (e214).

4.0 International (CC BY 4.0): https://creativecommons.org/licenses/by/4.0/], which permits unrestricted use, distribution, and reproduction in any

medium, provided the original author and source are credited. The official and authorized publication credit sources, which will be duly enforced, are

as follows: official journal title Amphibian & Reptile Conservation; official journal website: amphibian-reptile-conservation.org.

Received: 6 November 2018; Accepted: 5 August 2019; Published: 24 December 2019

The Asian snake genus Trimerodytes Cope, 1895

belongs to the family Natricidae and consists of four

species, namely Trimerodytes annularis (Hallowell,

1856), Trimerodytes percarinatus (Boulenger, 1899),

Trimerodytes aequifasciatus (Barbour, 1908), and

Trimerodytes yunnanensis (Rao and Yang, 1998).

Trimerodytes percarinatus (Boulenger, 1899) is a

nocturnal snake commonly known as the Eastern

Water Snake or Chinese Keelback Water Snake.

This species inhabits water passages in forested hilly

country (100—2,000 m asl) and generally feeds on fish,

crayfish, crustaceans, frogs, and their larvae (Pope 1935;

Smith 1943). Pope (1935) also recorded its presence

in irrigated fields near a forest in Kuatun, China.

Currently, 7rimerodytes percarinatus has two recognized

subspecies, Trimerodytes percarinatus percarinatus

(Boulenger, 1899) and Trimerodytes percarinatus suriki

(Maki, 1931).

Trimerodytes percarinatus was originally described as

Tropidonotus percarinatus by Boulenger in 1899, from

Kuatun, Foochow, in the north-west of the Province of

Fokien (=Fujian), China at an altitude of 3,000-—4,000

ft or more (Zhao and Adler 1993). Subsequently, it was

placed within the genus Natrix by Mell (1931) and later

assigned to the genus Sinonatrix by Rossman and Eberle

(1977). Based on a recent phylogenetic study, it 1s placed

within the genus 7rimerodytes (Ren et al. 2019).

This species is distributed in north-eastern India

(Arunachal Pradesh), Myanmar, Thailand, Laos,

Vietnam, south-eastern China, and Taiwan at elevations

ranging from 90-—2,000 m (Pope 1935; Smith 1943;

Taylor 1965; Zhao and Adler 1993; Stuart and Heatwole

2008; Nguyen et al. 2009; Boundy et al. 2014). Captain

and Patel (1998) first reported the existence of the genus

Trimerodytes in India, represented by this species,

7. percarinatus, based on an uncatalogued museum

specimen housed previously at Deban Forest Camp and

now at the Namdapha Tiger Reserve Field Museum in

Miao, India. For the next two decades since then, though

studies documenting the herpetofauna of Arunachal

Pradesh have been conducted, this species has not

been recorded (Athreya et al. 1998; Borang et al. 2005;

David and Mathew 2005; Agarwal et al. 2010). Here, we

report the first field record of 7rimerodytes percarinatus

(Boulenger, 1899) from India with notes on its natural

history.

This species was recorded on two occasions in

Namdapha Tiger Reserve, Arunachal Pradesh, India.

This national park harbors a rich biodiversity and is

part of one of the world’s biodiversity hotspots (Indo-

Myanmar). The first individual (Fig. 1) was encountered

at 27°28’58”N, 96°24’ 14”E and an elevation of 515 masl,

at approximately 2200 h on 17 June 2011. It was found

inside a small ditch filled with water (~1 ft deep) near

the road edge, with its head out of the water and it was

foraging actively. The second individual was recorded at

approximately 1730 h on 18 June 2011. It was foraging

in a water passage near the edge of a road. A Fowlea cf.

piscator was also seen foraging in the same water body.

The first individual was captured for morphological

measurements and photographs, and was later released

into the wild. A detailed description of the specimen is

given in Table 1, and follows the methods from Vogel et

al. (2004). Comparisons between the left and right sides

of the head, and associated habitat where the snake was

encountered, are also presented (Figs. 2-4).

Correspondence. ':*ashokgene@gmail.com, *isuvodee mail.com, * sangobarta@gmail.com, * ashokcaptain@hotmail.com

i 8! PUES if 8! [p

Amphib. Reptile Conserv.

December 2019 | Volume 13 | Number 2 | e214

Trimerodytes percarinatus in India

Fig. 1 Full body picture of Trimerodytes percarinatus from

Namdapha, India.

Fig. 3 Habitat of Trimerodytes percarinatus at Namdapha,

India.

Morphometric data for this specimen fall within

the range of Trimerodytes percarinatus as defined

in the available literature (Pope 1935; Smith 1943).

These observations provide two additional records of

Trimerodytes percarinatus from India and the first field

report from the country, though the sightings were in

more or less the same place (“Deban’”) associated with

the earlier specimen (see Captain and Patel 1998).

This work, as well as other literature on this genus

including new range records of other congeners from

Indochina (Vogel et al. 2004; Pauwels et al. 2009; Le

et al. 2015), points out our incomplete understanding of

the distribution of this genus as a whole. Further work

is required to determine the actual distribution range of

this species in India, to understand its morphological and

genetic variation across populations, and to add to our

knowledge of its natural history.

Acknowledgements.—We would like to thank the State

Forest Department of Arunachal Pradesh for providing

permission (No. CWL/G/13(17)/06-07/PT/3838-46)

to Kartik Shanker and Ashok Kumar Mallik, Centre

for Ecological Sciences, Indian Institute of Science,

Bangalore, to carry out the fieldwork and collect samples

Amphib. Reptile Conserv.

Fig. 2 Comparison between right and left sides of the head of

Trimerodytes percarinatus from Namdapha, India.

Fig. 4 Location of first field sighting of Trimerodytes percari-

natus in Namdapha Tiger Reserve, Arunachal Pradesh, India.

in protected areas of Arunachal Pradesh. We thank

Kartik Shanker for his valuable input to our manuscript,

and for providing financial and logistic support during

our fieldwork. Also, many thanks to the Field Director,

Assistant Field Director Dr. Aporesh Gupta-Choudhury,

and other forest staff from Deban guest house, Namdapha

Tiger Reserve, for their hospitality during the fieldwork.

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Pradesh, India: Notes on Some of the More Interesting

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PT. 2005. Checklist of the snakes of Arunachal

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Captain A, Patel A. 1998. Sinonatrix, a new genus for

India. Hamadryad 22(2): 114-115.

Cope ED. 1895. On a collection of Batrachia and

Reptilia from the Island of Hainan. Proceedings of the

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in Vietnam with a new country record of Sinonatrix

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169.

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Table 1. Characteristics of one specimen of 7rimerodytes percarinatus from Namdapha, Arunachal Pradesh, India, compared with

descriptions in Pope (1935) and Smith (1943).

Characters This study Pope (1935) Smith (1943)

Dorsal scale rows 19:19:17, keeled 19 19

Ventrals 153 138-143 133-157

Subcaudals 69/70 70—79 (males), 67-73 (females) 68-85

Anal Divided Divided -

vara 433 (male) 515-567 (males); 620-730 (females) 720 (male): 940 (female)

Tail length (mm) 131 130 190 (male); 270 (female)

Head length (mm) 22.1 — —

Horizontal eye 39 a =

diameter (mm)

Vertical eye diameter

3.9 w ,

(mm)

Eye to nasal distance 40 i 7

(mm)

Eye-snout distance

6.8 = -

(mm)

Inter-nasal distance 48 7 7

mm ;

Prefrontal (mm) 2.6 - —

Parietal (mm) 7.3 — —

Length of supraocular 53 _ 7

(mm)

Width of supraocular

27 a Ls

(mm)

Amphib. Reptile Conserv. 301

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Trimerodytes percarinatus in India

Table 1 (continued). Characteristics of one specimen of 7rimerodytes percarinatus from Namdapha, Arunachal Pradesh, India,

compared with descriptions in Pope (1935) and Smith (1943).

Characters This study Pope (1935) Smith (1943)

8 (left), 4 touching eye

1 th th 1

Supralabials /9 (right), 4" and 5 9 (rarely 8 or 10), 4 and 5" entering 9, 4" and 5” touching eye

the eye

touching eye

10 (on left and right);

1‘—5" touch anterior

genials (chin shields); 5 lower labials in contact with anterior

Infralabials 5 and 6" touch chin shields that are shorter than —

posterior genials — posterior

which are longer than

anterior genials

Preoculars 1 (+1 presubocular)/ 1 single 1

3 (+1 postsubocular/ 3

Postoculars (+2 postsuboculars) 4 (occasionally 3 or 5) 7

Supraocular 1/1 - _

1 (2 fused scales) + 3/ 1 3 (occasionally 2 or 4)/ 3

Temporals (2 fused scales) + 3 (occasionally 2, rarely 4) af oeratcly Ste)

Cross bands (on 30 = ls

body)

Nostril Directed upward — Directed slightly upward

Nasal Partially divided Completely divided —

Narrowed anteriorly, Natvowed anterior: donper than Distinctly narrowed anteriorly,

Internasal longer than the usually longer than the

broad, and longer than prefrontal

prefrontal prefrontal

Prefrontal Broader than long Shorter than internasal -

; As long as broad, as long as its

Frontal Long and pointed distance from the end of snout, shorter —

posteriorly than parietals

Young: dorsum grey or dark

Dorsum greyish brown

Olive green, color descending on

with uniform cross-bars,

doisolatcrlipontion _ Dorsum greyish olive, sides with the dorsolateral side as V shaped

lichtsyellowancolor light-edged black vertical bars; venter _bars, the interval between bars

Cuibeion otter Shieh ae uniform yellowish white anteriorly, and lower portion yellowish;

iaiformaparedand spotted and speckled with blackish Adults: dorsum greyish or

anpaned Pareles posteriorly; lower surface of tails dark —_ olivaceous with uniform or with

Withtlisht yellowish grey in color along with black spots dark reticulation or dark cross-

Sane eral bars, venter whitish, with or

without dark cross-bars

Amphib. Reptile Conserv. 302 December 2019 | Volume 13 | Number 2 | e214

Amphib. Reptile Conserv.

Mallik et al.

Ashok Kumar Mallik received his Doctorate degree in 2018 from the Centre for Ecological

Sciences, Indian Institute of Science, Bangalore, India. Ashok’s research interests include

systematics, taxonomy, hybrid zones and speciation, population genomics, and evolutionary

ecology of reptiles and amphibians. He is working on the systematics and biogeography of a few

genera of colubrid and viperid snakes in Peninsular India.

Subhadeep Chowdhury received his Bachelor’s degree in Zoology from Midnapore College,

India, in 2014 and his Master’s degree in Marine Biotechnology in 2016 from the Goa University,

India. Currently, Subhadeep is working independently on the herpetofauna of West Bengal

state. His current research interests include the natural history, biogeography, and systematics of

amphibians and reptiles of India.

Bharat Bhushan Bhatt received his Doctorate degree in 2004 from the Department of Zoology,

Guwahati University, Assam, India. Bharat is presently serving as Senior Scientific Officer in the

Department of Environment and Forest at the Office of the PCCF (Wildlife and Biodiversity),

Government of Arunachal Pradesh, India. He was one of the pioneers in pursuing an interest

in the field of herpetology in the state, and in North India as a whole. Bharat has more than 30

years of field experience in various wildlife subject matters. He has contributed to 13 published

scientific papers, and compiled many survey reports, case histories, and other contributions to the

state fauna series.

Ashok Captain is a renowned Indian herpetologist who is interested in and dabbles with the

traditional taxonomy of snake species that occur in India.

303 December 2019 | Volume 13 | Number 2 | e214

Official journal website:

amphibian-reptile-conservation.org

Amphibian & Reptile Conservation

13(2) [Special Section]: 304-322 (e215).

The most frog-diverse place in Middle America, with notes

on the conservation status of eight threatened species of

amphibians

12.* José Andrés Salazar-Zuniga, '?°Wagner Chaves-Acuna, Gerardo Chaves, ‘Alejandro Acuna,

12Juan Ignacio Abarca-Odio, '*Javier Lobon-Rovira, '?7Edwin Gomez-Méndez, ‘Ana Cecilia

Gutiérrez-Vannucchi, and 7Federico Bolafos

'Veragua Foundation for Rainforest Research, Limén, COSTA RICA *Escuela de Biologia, Universidad de Costa Rica, San Pedro, 11501-2060

San José, COSTA RICA Division Herpetologia, Museo Argentino de Ciencias Naturales “Bernardino Rivadavia”-CONICET, C1405DJR, Buenos

Aires, ARGENTINA ‘*CIBIO Research Centre in Biodiversity and Genetic Resources, InBIO, Universidade do Porto, Campus Agrario de Vairdo,

Rua Padre Armando Quintas 7, 4485-661 Vairdo, Vila do Conde, PORTUGAL

Abstract.—Regarding amphibians, Costa Rica exhibits the greatest species richness per unit area in Middle

America, with a total of 215 species reported to date. However, this number is likely an underestimate due to the

presence of many unexplored areas that are difficult to access. Between 2012 and 2017, a monitoring survey

of amphibians was conducted in the Central Caribbean of Costa Rica, on the northern edge of the Matama

mountains in the Talamanca mountain range, to study the distribution patterns and natural history of species

across this region, particularly those considered as endangered by the International Union for Conservation of

Nature. The results show the highest amphibian species richness among Middle America lowland evergreen

forests, with a notable anuran representation of 64 species. The greatest diversity in the study area occurred in

the mature forest on the basal belt. Of the 68 amphibian species found, seven (10%) are endemic to the Atlantic

versant and eight (11.6%) are threatened. This survey includes the first record of Gastrotheca cornuta in Costa

Rica since it was last reported 21 years ago. New populations of Agalychnis lemur (Critically Endangered)

and Duellmanohyla uranochroa (Endangered) are reported, and Ecnomiohyla veraguensis (Endangered) is

reported for the first time in Costa Rica. These findings show that this locality is a high priority conservation

area for a large number of amphibian species, which are often threatened by habitat loss and fragmentation.

Keywords. Biodiversity, Costa Rica, Endangered, Limon province, patterns of distribution, Tropical Wet Forest

Resumen.—En anfibios, Costa Rica exhibe la mayor riqueza de especies por unidad de area en América

Meridional con un total 215 especies documentadas a la fecha. Sin embargo, es probable que este numero este

subestimado debido a la presencia de areas inexploradas con dificil acceso. Entre 2012 y 2017, realizamos un

monitoreo de anfibios en el Caribe Central de Costa Rica, en el borde norte de la Fila Matama en la Cordillera

de Talamanca, para estudiar los patrones de distribucion y la historia natural de las especies en esta region,

particularmente aquellas consideradas en peligro por la Union Internacional para la Conservacion de la

Naturaleza (UICN). Nuestros resultados muestran la mayor riqueza de especies de anfibios en los bosques

perennes de tierras bajas de América Meridional, con una notable representacion de anuros de 64 especies.

La mayor diversidad en el area de estudio se encontro en el bosque maduro en el piso basal. Del total de

especies, siete (10%) son endemicas de la vertiente Atlantico y ocho (11,6%) estan amenazadas. Este es el

primer registro de Gastrotheca cornuta en Costa Rica después de 21 anos desde que se registro por ultima

vez. Descubrimos nuevas poblaciones de Agalychnis lemur (en Peligro Critico), Duellmanohyla uranochroa

(en Peligro), y reportamos por primera vez Ecnomiohyla veraguensis (en Peligro) en Costa Rica. Nuestros

resultados muestran que esta localidad es un area de alta prioridad para la conservacion de una gran cantidad

de especies de anfibios, a menudo amenazadas por la fragmentacion y la pérdida de habitat.

Palabras clave. Biodiversidad, Costa Rica, amenazado, provincia de Limon, patrones de distribucion, Bosque Trop1-

cal Humedo

Citation: Salazar-Zuniga JA, Chaves-Acuha W, Chaves G, Acuna A, Abarca-Odio JI, Lobon-Rovira J, Gdmez-Méndez E, Gutiérrez-Vannucchi AC,

Bolanos F. 2019. The most frog-diverse place in Middle America, with notes on the conservation status of eight threatened species of amphibians.

Amphibian & Reptile Conservation 13(2) [General Section]: 304-322 (e215).

tribution 4.0 International (CC BY 4.0): https://creativecommons.org/licenses/by/4.0/], which permits unrestricted use, distribution, and reproduction in

any medium, provided the original author and source are credited. The official and authorized publication credit sources, which will be duly enforced,

are as follows: official journal title Amphibian & Reptile Conservation; official journal website: amphibian-reptile-conservation.org.

Received: 21 March 2019; Accepted: 22 December 2019; Published: 30 December 2019

Correspondence. *jsalazar@veraguarainforest.com (JASZ), wchaves512@gmail.com (WCA), cachil3@gmail.com (GC),

alejandro2 lucr@gmail.com (AA), jiao24@gmail.com (JIAO), j.lobon.rovira@hotmail.com (JLR), gedgome@gmail.com (EGM),

anagv04@gmail.com (ACGV), federico. bolanos@ucr.ac.cr (FB)

Amphib. Reptile Conserv. 304 December 2019 | Volume 13 | Number 2 | e215

Salazar-Zuniga et al.

Introduction

Currently, more than 8,000 species of amphibians have

been described worldwide, with the greatest diversity

occurring in the Neotropics (Duellman 1999a; Frost

2019), where lower Central America stands out as a

region with a substantial number of species (Campbell

1999; Duellman 2001; Savage 2002; Kubicki 2007).

However, there has been an increase in the numbers

of species designated as endangered throughout this

region since the late 1980s due to habitat deforestation

(Young et al. 2001; Stuart et al. 2004; Becker et al. 2007),

climate change (Pounds et al. 1999; Hof et al. 2011), and

infectious diseases (Lips et al. 2003; Pounds et al. 2006;

Wake and Vredenburg 2008).

Tropical forests harbor a considerable number

of amphibian species across distinct microhabitats

that are often related to water-dependent sites such

as ponds, temporary swamps, streams, tree holes,

and bromeliad axils (Duellman 1970; Savage 2002;

Lehtinen et al. 2004; Haddad and Prado 2005).

However, many other amphibians exhibit reproductive

modes that are totally independent from water bodies

(Savage 2002). For instance, frogs of the genera

Gastrotheca (Hemiphractidae), Eleutherodactylus

(Eleutherodactylidae), /ncilius (formerly Crepidophryne:

Bufonidae), Craugastor, Pristimantis, and Strabomantis

(Craugastoridae) lay encapsulated eggs out of water, in

which embryos undergo direct development and hatch

out as small adults (Savage 2002; Gray and Bland 2016).

Costa Rica is known to possess considerable

amphibian species richness per unit area (Savage 2002;

Sasa et al. 2010; Bolafios et al. 2011). However, this

great richness is likely to be underestimated due to the

presence of undiscovered and undescribed species in

areas that tend to be not easily accessible. To date, several

previous works have reported countrywide amphibian

checklists (Savage 2002; Bolafios et al. 2011; Leenders

2016). In particular, an increasing interest has focused

on documenting species occurrence and the population

status of threatened species in Costa Rica’s montane

ecosystems (Hayes et al. 1989; Abarca 2012; Acosta-

Chaves et al. 2015; Rovito et al. 2015) and tropical forests

of varying altitudinal gradients in the Pacific Slope

throughout its South (McDiarmid and Savage 2005),

Central (Laurencio and Malone 2009), and Northern

regions (Sasa and Solorzano 1995). In contrast, the

amphibian diversity of the Costa Rican Atlantic has been

broadly documented almost exclusively in its Northern

region (Donnelly and Guyer 1994; Guyer and Donnelly

2005; Whitfield et al. 2007). Only recently, Kubicki

(2008) compiled the first list of species in premontane

moist forests of the Costa Rican central-Caribbean.

Although that inventory showed the relevant diversity of

amphibians, little is known about the current population

status and distribution of amphibian species across other

areas of the mid-Caribbean.

Amphib. Reptile Conserv.

Along this region, the Talamanca mountain range

stands out as a predicted high priority conservation

area for amphibians (Garcia-Rodriguez et al. 2011).

Considering that long-term monitoring can accurately

assess population conditions (La Marca et al. 2005),

species inventories are key to covering gaps in the

distribution and natural history of endangered species in

order to determine appropriate conservation strategies

(Peloso 2010; Verdade et al. 2012). This study assessed

the local richness and species distribution of amphibians

in Veragua Rainforest Eco Research and Adventure Park

and its surroundings on the northern edge of the Matama

mountains in the Talamanca mountain range. Diversity

analysis was conducted for different types of altitudinal

belts, forests, and microhabitats found across the

sampling area, and the population status of the threatened

Species in the study area are discussed.

Materials and Methods

Study area. The study was conducted in the Central

Caribbean of Costa Rica between Las Brisas de Veragua

town (9°57°07"°N, 83°12711”E; 233 m asl) and Platano

peak (9°51’50”N, 83°14°10”E; 1,000 m asl) on the

northern edge of the Matama mountains in the Talamanca

mountain range, including Chimu peak (9°52’48’N,

83°14°13”E; 741 m asl) and Veragua Rainforest Park

(VRP; 9°55’30’N, 83°11’28”E; 420 m asl; Fig. 1). This

private reserve covers 3,200 ha of protected land ranging

from 200-420 m asl, and it comprises mature forest,

secondary vegetation at different stages of regeneration,

open areas, and dirt roads. The study site lies adjacent to

Victoria (9°55’21.73”N, 83°10’2.43”E; 410 m asl) on the

Victoria river basin and the Matama mountains. This area

is the closest point of the Talamanca mountain range to

the Caribbean Sea and it forms part of the buffer zones

of La Amistad International Park (an UNESCO World

Heritage Site), the Banano river basin protected area, and

the influence zones of the Zent, Peje, and Chirrip6 rivers,

as well as the Bajo Chirrip6 indigenous reserve (SINAC

2018). Sampling was carried out along the elevation

range 200—1,000 m asl, where two types of forest are

located according to Holdridge (1967): Basal Tropical

Wet Forest (200-600 m asl) and Premontane Tropical

Wet Forest (601—1,000 m asl). Only these altitudinal

belts are recorded and reported here, because they both

represent the Tropical Wet Forest.

Data collection. Data were collected between January

2012 and December 2017. To record the species

richness, samplings were standardized through diurnal

and nocturnal visual and acoustic recognition searches

(Crump and Scott 1994) into three transects: two in

VRP, covering approximately 4 km each (Transects A

and B), and one carried out along an 11 km trail between

the reserve and Platano peak (Transect C), including

Chimu peak halfway along the route. On each of these

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Extreme frog diversity in Costa Rica

i on NICARAGUA

EES National Park “Barbilla”

/] National Park “La Amistad”

Indigeous Reserve “Chirripoi”

tT Indigeous Reserve “Bajo Chirripoi”

Fig. 1. Location of the study area (51 km?) in the Limon Province in the Central Caribbean area of Costa Rica. The colored points

represent the main localities of the study area: yellow (Chimt Peak), red (Platano Peak), blue (Veragua Rainforest Research and

Adventure Park), green (Las Brisas), and purple (Victoria).

transects, surveyors walked side-by-side at a constant

speed to record amphibian diversity on both sides of

the trail (Seber 1986), covering up to 10 m from each

side towards the forest. Transect A (300—420 m asl) was

located along a dirt road inside the reserve in a secondary

forest edge that included a 30 m wide natural pond and

open areas. Transect B (200-400 m asl) covered forest

trails and riparian environments within a mature forest.

Transect C (400—1,000 m asl) comprised an old wood

road (4 km) in a secondary forest and an indigenous

trail (7 km) within a pristine environment that included

natural ponds and riparian habitats along the trail.

From January 2012 to December 2012, Transect A

or B was sampled weekly during the day (6:00—11:00

h) and at night (18:00—22:00 h), totaling 27 field days

with four person hours (ph) per transect for a total search

effort of 1,728 ph. Once a year, between 2012 and 2017,

six expeditions at Transect C were conducted, totaling

13 field days of diurnal and nocturnal monitoring and a

search effort of 1,560 ph. A leaf litter plot survey (Scott

1976) was used to sample ten plots (8 x 8 m) in Chimt

peak (2014, 2017) and Platano peak (2013, 2015) on an

annual basis, for a total of 40 sampled plots.

The following information was recorded for the species

detected during monitoring: 1. Holdridge altitudinal belts:

basal (b) or premontane (p); 2. Type of forest: mature

(M) or disturbed (D; includes secondary forest and open

areas), and 3. Habitat association: riparian (R), forest (F),

or swamp (S; including temporary or permanent ponds).

If possible, one specimen per species was collected on

each Holdridge life zone. The collected specimens were

anesthetized and euthanized with lidocaine, fixed in a

10% buffered formalin solution, and later preserved in

70% ethanol solution. For all specimens, tissue samples

Amphib. Reptile Conserv.

of muscle and liver were collected and fixed in 95%

ethanol. Voucher specimens and tissue samples were

deposited at the Museo de Zoologia of the Universidad

de Costa Rica (UCR). Some specimens were collected by

third parties or other VRP researchers through occasional

encounters in random field trips.

The species list includes the information obtained from

this monitoring effort and UCR records of the Victoria

locality, covering a study area of 51 km/?, hereafter

referred to as Veragua. Additional photographic material

from collaborations with specialists in this area was

evaluated. The taxonomic nomenclature follows Frost

(2019), except for hylids in which Faivovich et al. (2018)

was followed. The conservation status of each species

was categorized according to the Red List of Threatened

Species of the International Union for Conservation of

Nature (IUCN 2019) and registered observations on the

natural history of threatened species.

Data analysis and permission. The Jaccard index (],) was

used to determine the similarity in species composition

between altitudinal belts, forest types, and habitat associa-

tion. A species accumulation curve was performed to ac-

count for species richness. Sampling was conducted under

research permit SINAC-ACLAC-PIME-VS-R-024-2016,

granted by Sistema Nacional de Areas de Conservacion

(National System of Conservation Areas, SINAC).

Results

Overall Results

The surveys recorded a total of 68 species of

amphibians, including 64 anurans distributed in 11

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Salazar-Zuniga et al.

Number of species

2010 2011 2012 2013 2014 2015 2016 2017

Year

Fig. 2. Amphibian species accumulation curve for the 2010-

2017 period in the study area.

families and 31 genera, three salamanders of the family

Plethodontidae in two genera, and one caecilian in the

family Caeciliidae (Table 1; Plates I-V). The most

speciose families were Hylidae with 22 species (32.4%),

followed by Craugastoridae with 15 species (21.7%) and

Centrolenidae with 10 species (14.5%) [Table 1]. Six

of the 68 species (8.7%) were endemic to the Atlantic

slope of Costa Rica: Bolitoglossa alvaradoi, Oedipina

berlini, Craugastor persimilis, Diasporus amirae,

Hyalinobatrachium dianae, and Ecnomiohyla sukia. The

species accumulation curve reached an asymptotic phase

at the end of the sampling period (Fig. 2).

Low species similarity was obtained between basal

and premontane belts (i = 0.37) and the majority

of premontane species were found in the basal belt,

except for C. persimilis, D. amirae, and Pristimantis

caryophyllaceus (Table 1; Fig. 3a). Mature forests and

disturbed areas were found to share slightly more than

half of the species (I, = 0.52). A total of 19 species were

only present in mature forests, and 14 species were

detected only in disturbed areas (Table 1; Fig. 3b). The

most diverse habitats were the forest (43 sp.) and riparian

(31 sp.) environments, while 20 species were associated

with swamps (Fig. 3c; Table 1). The results show that

44 species were only found in one type of habitat; out

of these, the forest (19 sp.) was the most diverse habitat,

followed by riparian environments (16 sp.) and swamps

(9 sp.; Table 1). A medium-low similarity was found in

the composition of species between the riparian and the

forest (15 sp.; I; = 0.25), as well as between the swamps

and the forest (11 sp.; I; =0.21), although the data indicated

only a minimal similarity when comparing rivers and

swamps (2 sp.; I, = 0.04; Table 1). The only species that

were found in all three habitats were Agalychnis spurrelli

and Rhinella horribilis.

According to the IUCN conservation status, one

species is categorized as Data Deficient (DD), 54 as

Least Concern (LC), and eight in the various threatened

categories. Pristimantis altae and P. caryophyllaceus

are categorized as Near Threatened (NT); Craugastor

persimilis as Vulnerable (VU); Duellmanohyla

uranochroa, Ecnomiohyla_ veraguensis, Gastrotheca

cornuta, and Bolitoglossa alvaradoi as Endangered

Amphib. Reptile Conserv.

Number of species

es)

ro)

Basal Premontane

Altitudinal Belt

Lv)

Number of species

Mature Disturbed

Forest Type

Number of species

Forest

Habitat

Fig. 3. Number of species registered according to the altitudinal

belt (A), forest type (B), and habitat (C) in the study area.

Riparian Swamp

(EN); and Agalychnis lemur as Critically Endangered

(CR; Table 1). The species Hyalinobatrachium dianae,

D. amirae, C. sylviae, E. sukia, Ecnomiohyla bailarina,

and O. berlini remain uncategorized (Table 1).

Regarding the uncategorized species populations,

few populations of H. dianae were observed in isolated

streams within the basal mature forest. Populations with

several individuals of Diasporus amirae were detected

at the premontane belt. The species C. sy/viae was found

during 2011—2012 in only a few places within the forest.

Generally, the adults were observed inside tree holes

up to 3 m high. However, between 2012 and 2017, the

species was more commonly observed reproducing

throughout the year in VRP, near small artificial ponds

(length 200 cm, width 150 cm, depth 50 cm) located

within the forest. These ponds were created in 2012 by

a project of the Veragua Foundation for the Rainforest

Research “Veragua Foundation” (NGO) that aims to

establish in situ breeding sites for the conservancy and

study of the native amphibians. Ecnomiohyla bailarina

and E. sukia were detected calling from the canopy in

basal and premontane pristine forest. Oedipina berlini

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Extreme frog diversity in Costa Rica

Table 1. Checklist of amphibians of the Veragua Rainforest Eco Research and Adventure Park and its surroundings, with information

on the voucher ID (UCR), IUCN status, altitudinal belt (basal [b] / premontane [p]), forest type (mature [M] / disturbed [D]), and

habitat association (forest [F] / swamp [S] / riparian [R]).

Taxa UCR IUCNstatus Altiudinal Belts Forest Types Habitats

Aromobatidae (1)

Allobates talamancae (Cope, 1875) 21593 LC bp MD FS

Bufonidae (4)

Incilius coniferus (Cope, 1862) 21164 Le b MD FS

Incilius melanochlorus (Cope, 1877) 21983 Le bp M RF

Rhaebo haematiticus Cope, 1862 21139 Ee bp MD RF

Rhinella horribilis (Wiegmann, 1833) 21148 be b D RFS

Centrolenidae (10)

Cochranella granulosa (Taylor, 1949) 23183 Le b MD R

Espadarana prosoblepon (Boettger, 1892) Le b D R

Hyalinobatrachium chirripoi (Taylor, 1958) 21431 Le bp MD R

Hyalinobatrachium dianae Kubicki, Salazar, and 22035 b M R

Puschendorf, 2015

Hyalinobatrachium fleischmanni (Boettger, 1893) 23182 LC b MD R

Hyalinobatrachium talamancae (Taylor, 1952) 21157 LC bp M R

Hyalinobatrachium valerioi (Dunn, 1931) 21140 LG. b M R

Sachatamia albomaculata (Taylor, 1949) 21114 Le bp MD R

Teratohyla pulverata (Peters, 1873) 21153 LC b MD R

Teratohyla spinosa (Taylor, 1949) 21126 Le b MD R

Craugastoridae (15)

Craugastor brandsfordi (Cope, “1885,” 1886) 21149 LC b D F

Craugastor crassidigitus (Taylor, 1952) 21120 Le bp MD RF

Craugastor fitzingeri (Schmidt, 1857) 21150 Le bp MD F.

Craugastor gollmeri (Peters, 1863) 22550 LC bp M F

Craugastor megacephalus (Cope, 1875) ie b M E

Craugastor mimus (Taylor, 1955) 21414 Le b MD FP

Craugastor noblei (Barbour and Dunn, 1921) 21156 LC b MD F

Craugastor persimilis (Barbour, 1926) 22529 VU b M F

Craugastor polyptychus (Cope, 1886) 21121 LC bp MD F

Craugastor talamancae (Dunn, 1931) Le b D RF

Pristimantis altae (Dunn, 1942) 21145 NT bp MD RF

Pristimantis caryophyllaceus (Barbour, 1928) 21844 NT p M E

Pristimantis cerasinus (Cope, 1875) 21127 Le bp MD F

Pristimantis cruentus (Peters, 1873) 21170 Le bp M RF

Pristimantis ridens (Cope, 1866) 21096 LC bp MD RF

Dendrobatidae (4)

Dendrobates auratus (Girard, 1855) 21128 LC b MD F

Oophaga pumilio (Schmidt, 1857) 21106 Le bp MD RF

Phyllobates lugubris (Schmidt, 1857) 21143 Le b MD RF

Silverstoneia flotator (Dunn, 1931) 21986 Le bp MD RF

Eleutherodactylidae (2)

Diasporus diastema (Cope, 1875) 21415 Le bp MD RF

Diasporus amirae Arias, Chaves, Salazar, Salazar- 22010 p M F

Zufiga, and Garcia-Rodriguez, 2019

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Salazar-Zuniga et al.

Table 1 (continued). Checklist of amphibians of the Veragua Rainforest Eco Research and Adventure Park and its surroundings,

with information on the voucher ID (UCR), IUCN status, altitudinal belt (basal [b] / premontane [p]), forest type (mature [M] /

disturbed [D]), and habitat association (forest [F] / swamp [S] / riparian [R]).

Taxa UCR IUCN status Altiudinal Belts Forest Types Habitats

Hemiphractidae (1)

Gastrotheca cornuta (Boulenger, 1898) 21017 EN bp M R

Hylidae (23)

Agalychnis callidryas (Cope, 1862) 21098 1, & bp MD FS

Agalychnis lemur (Boulenger, 1882) 21104 CR bp MD S

Agalychnis saltator Taylor, 1955 21429 1 @ b MD FS

Agalychnis spurrelli (Boulenger, 1913) 21119 Le b MD RFS

Boana rufitela (Fouquette, 1961) Le b M S

Cruziohyla sylviae Gray, 2018 21422 b MD FS

Dendropsophus ebraccatus (Cope, 1886) 21103 Le b MD

Dendropsophus phlebodes (Stejneger, 1906) 21432 | D

Duellmanohyla rufioculis (Taylor, 1952) Le bp M R

Duellmanohyla uranochroa (Cope, 1875) 22002 EN bp M RF

Ecnomiohyla bailarina Batista, Hertz, Mebert, 22287 b M F

Kohler, Lotzkat, Ponce, and Vesely, 2014

Ecnomiohyla sukia Savage and Kubicki, 2010 22940 bp M

Ecnomiohyla veraguensis Batista, Hertz, Mebert, 21941 EN bp

Kohler, Lotzkat, Ponce, and Vesely, 2014

Hyloscirtus palmeri (Boulenger, 1908) 21995 LG b MD R

Isthmohyla lancasteri (Barbour, 1928) 21994 LC b M R

Scinax boulengeri (Cope, 1887) Le b D S

Scinax elaeochroa (Cope, 1875) 21151 LC b D FS

Smilisca manisorum (Taylor, 1954) Le b D S

Smilisca phaeota (Cope, 1862) 21113 Le b MD S

Smilisca puma (Cope, 1885) LC b D S

Smilisca sordida (Peters, 1863) 21099 Le b MD R

Tlalocohyla loquax (Gaige and Stuart, 1934) 21097 Le b MD S

Leptodactylidae (2)

Leptodactylus melanonotus (Hallowell, 1861) 21518 Le b D FS

Leptodactylus savagei Heyer, 2005 20107 Le b MD FS

Microhylidae (1)

Hypopachus pictiventris (Cope, 1886) Le b D SF

Ranidae (2)

Lithobates vaillanti (Brocchi, 1877) 21101 LC b D R

Lithobates warszewitschii (Schmidt, 1857) 21102 LE b MD RF

Plethodontidae (3)

Bolitoglossa alvaradoi Taylor, 1952 22048 EN b M

Bolitoglossa colonnea (Dunn, 1924) 21178 LE bp MD

Oedipina berlini Kubicki, 2016 22882 b D F

Caeciliidae (1)

Caecilia volcani Taylor, 1969 DD b D F

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Extreme frog diversity in Costa Rica

ie ee VE

Plate I. Photos in life o

bag

aie

f amphibians recorded in the sampling area: (A) A//obates talamancae; (B) Incilius coniferus, (C) I.

melanochlorus, (D) Rhaebo haematiticus, (E) Rhinella horribilis, (F) Cochranella granulosa, (G) Espadarana prosoblepon, (H)

Hyalinobatrachium chirripoi; (1) H. dianae; (J) H. fleischmanni, (IX) H. talamancae; (L) H. valerioi; (M) Sachatamia albomaculata;

(N) Teratohyla pulverata, and (O) T: spinosa. Photos by Victor Acosta-Chaves (C, G, N); Javier Lobon-Rovira (A, F, L, M, O); José

Andrés Salazar-Zuniga (B, D, E, H, J, K); Andréi Solis (1).

was observed once in the secondary basal forest, as an

individual was on the leaf litter on one of the trails of the

Veragua Rainforest Park at night.

Observations on the Threatened Species

Pristimantis altae (NT; Plate II-K) was seen and heard in

mature and secondary forests (Table 1). At the basal belt,

Amphib. Reptile Conserv.

isolated males were recorded on riversides and inside

the forest; nevertheless, at the premontane belt, groups

of at least six calling males were registered, found very

close to each other (2-3 m apart). At the premontane

belt, P. caryophyllaceus (NT; Plate H-L) was commonly

observed perched on leaves in the understory (100-150

cm high). On one occasion, a female was found in

brooding position over a fully developed clutch with 26

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Salazar-Zuniga et al.

Plate II. Photos in life of amphibians recorded in the sampling area: (A) Craugastor brandsfordi; (B) C. crassidigitus, (C)

C. fitzingeri, (D) C. gollmeri;, (E) C. megacephalus, (F) C. mimus; (G) C. noblei; (H) C. persimilis; (I) C. polyptychus, (J) C.

talamancae, (KK) Pristimantis altae, (L) P. caryophyllaceus, (M) P. cerasinus; (N) P. cruentus, (O) P. ridens. Photos by Victor

Acosta-Chaves (C, F); Javier Lobon-Rovira (B, K, O); José Andrés Salazar-Zurtiga (A, D, E, G-J, L—-N).

eggs inside a partially rolled leaf at 1 m high. The female

and her eggs were collected and placed in a plastic bag,

and the next day all the eggs had hatched inside the bag.

Craugastor persimilis (VU; Plate II-H) was observed

several times during the plot survey, hidden in the leaf

litter in the premontane belt.

Duellmanohyla uranochroa (EN; Plate IV-E) was

detected in a few streams in pristine forest (Table

1). Males were observed calling from the streamside

Amphib. Reptile Conserv.

vegetation. Also, a male chorus was heard inside the forest

at a distance of at least 100 m from the nearest stream.

Some of these individuals were located in between the

aerial roots of a walking palm (Socratea exorrhiza).

Ecnomiohyla veraguensis (EN; Plate 1V-H) was observed

once calling from the canopy in the basal mature forest.

Gastrotheca cornuta (EN; Plate III-G) was uncommon in

the survey samplings. Only two populations are known

in the study area; one of them was last reported in 1996

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Extreme frog diversity in Costa Rica

| \ By, ha . ee 5 }

Plate III. Photos in life of amphibians recorded in the sampling area (unless otherwise specified, all photographs refer to specimens

detected in Veragua): (A) Dendrobates auratus, (B) Oophaga pumilio, (C) Phyllobates lugubris, (D) Silverstoneia flotator, (E)

Diasporus diastema, (F) Diasporus amirae; (G) Gastrotheca cornuta (Veraguas, Panama); (H) Agalychnis callidryas, (1) A. lemur;

(J) A. saltator;, (IK) A. spurrelli, (L) Boana rufitela. Photos by Abel Batista (G); Javier Lobon-Rovira (A-C, E, H, I, K, L); José

Andrés Salazar-Zutiga (D, F, J).

at 200 m asl in the Victoria river basin (Solorzano et al.

1998), and the subsequent record was published 16 years

later, in the streamside vegetation in a deep canyon of the

Zent River at 550 m asl (Salazar 2015).

Three populations of the Critically Endangered

Agalychnis lemur (Plate III-[1) were observed throughout

Veragua. One of the populations was located in a pond

(width 35 m) in a mature premontane forest, where

many males were observed calling from the vegetation

at 50-150 cm high. The other two populations were

found in secondary forest at the basal belt, including a

population that is on the border of the VRP (Table 1).

Outside the reserve, this species was observed in small

ponds and flooding banks next to a wood extraction

road. Thus, given this immediate threat, small artificial

ponds (length 200 cm, width 150 cm, depth 50 cm)

were created by the Veragua Foundation during 2015

Amphib. Reptile Conserv.

to protect this population. As of 2016, it was easier to

observe individuals throughout the year during high-

humidity nights near these reproductive sites.

Discussion

With 215 species, Costa Rica is the 19™ richest country

in the world for amphibians, and exhibits the highest

richness per unit area in Middle America (Kubicki 2008;

Frost 2019). In this region, amphibian species density

seems to be greater towards the South, specifically in

Costa Rica and Panama (Campbell 1999). The results

presented here show that Veragua exhibits the highest

known species richness among Middle American lowland

evergreen forests. With a notable anuran representation of

64 species in 51 km? surveyed (Table 1; Plates I-V), these

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Salazar-Zuniga et al.

Plate IV. Photos in life of amphibians recorded in the sampling area (unless otherwise specified, all photographs refer to specimens

detected in Veragua): (A) Cruziohyla sylviae; (B) Dendropsophus ebraccatus, (C) D. phlebodes; (D) Duellmanohyla rufioculis; (E)

D. uranochroa,; (F) Ecnomiohyla bailarina (Darien, Panama); (G) E. sukia; (H) E. veraguensis (Veraguas, Panama); (1) Hy/oscirtus

palmeri; (J) Isthmohyla lancasteri;, (KK) Scinax boulengeri;, (L) S. elaeochroa, (M) Smilisca manisorum, (N) S. phaeota; (O) S.

puma, (P) S. sordida; (Q) Tlalocohyla loquax. Photos by Victor Acosta-Chaves (M); Abel Batista (F), Edwin Gomez-Méndez (O);

Daniel Hernandez (C); Andreas Hertz (H); Javier Lob6n-Rovira (A, B, J, N); José Andrés Salazar-Zutiga (D, E, G, I, K, L, P, Q).

Amphib. Reptile Conserv. 313 December 2019 | Volume 13 | Number 2 | e215

Extreme frog diversity in Costa Rica

Plate V. Photos in life of amphibians recorded in

ald -

melanonotus: (B) ib. savagei: (C) Hypopachus

pictiventris; (D) Lithobates vaillanti;, (E) L. warszewitschii; (F) Bolitoglossa alvaradoi, (G) B. colonnea, (H) Oedipina berlini; (1)

Caecilia volcani. Photos by Victor Acosta-Chaves (C, D, F); Esmeralda Arévalo (1); Javier Lobon-Rovira (E); José Andrés Salazar-

Zuniga (A, B, G, H).

surveys reveal one of the highest numbers of amphibian

species reported per unit area in the Neotropics (Savage

2002; Boza-Oviedo et al. 2012; Barrio-Amoros et al.

2011; Hertz et al. 2012; Arias and Bolafios 2014; Ferreira

et al. 2017).

In comparison with the most important diversity

hot spots from South America, the richest region of

amphibian species worldwide (Ron et al. 2018; Frost

2019), Veragua is also one of the most diverse localities

in the Neotropics with 68 species. Only certain sites

across the Amazon lowlands exhibit a greater richness

of amphibians than these Veragua sites (Barrio-Amor0os

et al. 2011; Ferreira et al. 2017). For example, amphibian

richness in Brazil ranged from 18 species (Alter do Chao,

Para) to as many as 78 species along a small section of

the Jurua river (Zimmermann and Rodrigues 1990; Lima

2008; Queiroz et al. 2011; Pereira-Junior et al. 2013;

Araujo and Costa-Campos 2014; Alves-Binicio and

Dias-Lima 2017; Ferreira et al. 2017; Lima et al. 2017),

with the extreme exception of 109 amphibian species in

the middle of the Xingu River (Vaz-Silvia et al. 2015;

Ferreira et al. 2017). In Peru, the most diverse sites are

in Bajo Rio Llullapichis (74 sp.; Schluter et al. 2004),

Parque Nacional Manu (68 sp.; Morales and Mcdiarmid

1996), and Cuzco Amazonico (64 sp.; Duellman 2005;

Barrio-Amoros et al. 2011). In Colombia, the highest

diversity was found in Leticia 97 species (Lynch 2005),

Amphib. Reptile Conserv.

and the two most important inventories reported in

Ecuador are from the village of Santa Cecilia (87 sp.;

Duellman 1978) and Parque Nacional Yasuni (135 sp.),

currently the most diverse amphibian site in the world

(Ron et al. 2018).

In Costa Rica, the amphibian richness is concentrated

in the southern lowlands of the country and in the

northeastern Atlantic versant (Campbell 1999;

McDiarmid and Savage 2005; Santos-Barrera et al.

2008). Compared to the three other major inventories

in these areas, Veragua is more species-rich than either

the South Pacific locality of Rincon (47 sp., Anura [42]/

Caudata [4]/Gymnophiona [1]; McDiarmid and Savage

2005), the Atlantic versant sites of La Selva Biological

Station (52 sp., 47/3/1; Guyer and Donnelly 2005),

or Guayacan (66 sp., 58/6/2; Kubicki 2008). Other

important inventories were reported in a transitional wet-

dry forest in the locality of Carara (39 sp.; Laurencio and

Malone 2009), and among the richest sites in dry forests

is Finca Taboba, at the northern edge of the country (21

sp.; Campbell 1999).

The low diversity of salamanders found in Veragua

could be due to fact that the sampling area was below

1,100 m asl, and that only one monitoring was conducted

per year in the premontane belt. It is also possible that

the sampling method was not inclusive enough to cover a

broader diversity in the basal belt. The number of frog and

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Salazar-Zuniga et al.

salamander species distributed along specific elevations

in Middle America follows a pattern previously noted in

Guatemala and Belize, 1.e., a moderate number of species

in the lowlands that progressively increases as it reaches

moderate or intermediate elevations, and then declines

precipitously at higher elevations (Campbell and Vannini

1989; Wells 2007). Salamanders partly show this pattern

as they exhibit a more dramatic increase in species in the

highlands (Campbell 1999; Wells 2007). We presume

that some other species, such as Bolitoglossa striatula,

Nototriton matama, Oedipina carablanca, and Oedipina

gracilis, which occur in the Atlantic Versant at a similar

altitudinal belt and close to the Talamanca mountain range

(Savage 2002; Kubicki 2008; Leenders 2016), could

also occur in Veragua. Among caecilids, only Caecilia

volcani was found. The apparent low diversity in this

group is probably caused by its fossorial habits (Peloso

2010; Ferreira et al. 2017) which undermine effective

sampling. We think that at least one common caecilid

species in the Atlantic Versant (Gymnophis multiplicate)

could be present in Veragua (Leenders 2016).

In this study, the basal belt was found to be more

diverse than the premontane (Fig. 3; Table 1). This is a

generalized pattern of distribution among anurans (Savage

2002) and should account for the vast representation of

frogs and toads in this study, which represent 94% of the

total amphibian species. The latter species distribution is

similar to that of the wet slopes of the Andes, the region

with the highest diversity of anuran species in the world

(Duellman 1999b; Wells 2007). The higher number

of anuran species in old mature forests compared with

disturbed areas might be explained by the presence of

more microhabitats in pristine environments (Table 1;

Savage 2002; Acosta-Chaves et al. 2015). Nevertheless,

one of the main factors shown to influence high local

and regional diversity is the variety of habitats with

numerous vegetation types, ranging from forests to open

grasslands, that occur side-by-side in the landscape, each

of them harboring a different array of species (Colli et al.

2002; Nogueira et al. 2009; Lopes-Santos et al. 2014).

The greatest species diversity and the highest level

of endemism for amphibians in Middle America occur

along the windward mesic slopes of major mountain

ranges between elevations of 800 and 2,800 m asl,

which in Costa Rica include the Guanacaste, Tilaran,

and Talamanca mountain ranges (Campbell 1999). The

Talamanca mountain range is recognized as a site of

speciation and a dispersion center for several species with

a high degree of endemism (Arias and Bolafios 2014).

Among amphibians and reptiles, 27% of the species in

Costa Rica are endemic to this region (Campbell 1999;

Savage 2002; Chaves et al. 2009; Streicher et al. 2009;

Boza-Oviedo et al. 2012; Arias and Bolafios 2014). In

this study, 8.7% of the amphibians are endemic to the

Atlantic Versant of Costa Rica (Campbell 1999; Leenders

2016; Frost 2019), including species of the genera

Duellmanohyla and Isthmohyla, which are endemic to

Amphib. Reptile Conserv.

Middle America (Faivovich et al. 2018).

A high number of species was found along the forest

and the riparian habitats (Fig. 3; Table 1). This association

with a type of forest might be due to the great variety of

microhabitats found throughout these environments that

result from irregular topography (Wells 2007; Kubicki

2008). Leaf litter is an important habitat for anurans,

especially among species with terrestrial reproduction

(although it is not restricted to them; Wells 2007). The

high moisture levels found in the forest floor allow

terrestrial species to forage and call during either the day

or night (Wells 2007). The species richness of leaf litter

frogs and toads is positively correlated with the number

of wet months and the litter mass depth (Wells 2007;

Whitfield et al. 2007).

The distribution along the riparian habitats was found

to be non-uniform, as it could rely on vegetation coverage

and the physical characteristics of the environment. The

high number of amphibian species found along riparian

habitats in this study could be due to the numerous springs,

streams, and torrents found throughout the sampling area

(Fig. 1), which generate several microhabitats along this

environment (McDiarmid and Savage 2005; Kubicki

2007; SINAC 2018). Previous studies found a similar

distribution pattern in Guayacan, where 35% of the

Species are associated with lotic environments (Kubicki

2008), while glass frogs represent more than 16% of the

amphibian diversity in La Selva (Guyer and Donnelly

2005) and Rincon (McDiarmid and Savage 2005). In the

mountainous regions of Middle America, the permanent

or temporary ponds required for amphibian breeding

are often scarce (Wells 2007). This pattern is also seen

at Veragua and Guayacan, where the highly irregular

topography of these localities causes permanent ponds

to be a much more limited resource for reproduction

(Kubicki 2008). Nevertheless, pond-breeders account

for a notable representation of anuran species in Veragua

(32, 4%) and Guayacan (30%; Kubicki 2008).

The species accumulation curve reached an asymptote,

meaning that the sampling effort to detect species

produced a number near the maximum expected value

(Fig. 2). Nevertheless, there may be some cryptic species

groups with high variation, and this fact may obscure the

estimates given here, as different species could be hidden

under a single name (Funk et al. 2012; Alves-Binicio

and Dias-Lima 2017). However, the clarification of such

unknown diversity requires further integrative taxonomic

studies. Likewise, we suggest a larger survey effort in the

premontane belt, since the difficult access to sampling

areas did not allow for a continuous sampling.

Comments on Threatened Species

This study registered P altae (NT), which has been

previously reported in very few places on the Atlantic

Slope of Costa Rica and northwestern Panama (Leenders

2016). Overall, there is only limited information about

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Extreme frog diversity in Costa Rica

its natural history, population trends, and conservation

needs (Leenders 2016). Historically, P. altae has been

associated with undisturbed areas (Savage 2002; Pounds

et al. 2008a). Even though this species was observed in

mature forests, it was also commonly detected close to

streams within secondary forests. According to Savage

(2002), this species is mute; however, during high

humidity dark nights it is common to hear the species

emitting a short two note call “clock-clock,” similar

to the sound that two glass marbles emit when they hit

each other twice very rapidly. In basal secondary forests,

isolated males were heard calling in the forest. However,

on the premontane belt, this species is more abundant,

and several males were heard calling at a close distance

from each other. Unfortunately, the calls are emitted

sporadically, and have not yet been recorded.

In this study, P. caryophyllaceus (NT) was one of

the most common frogs in the mature premontane belt.

During the mid-1980s, when many populations in Costa

Rica declined (Leenders 2016), P. caryophyllaceus

disappeared from most lowlands; however, it persisted

at higher elevations (Leenders 2016). This pattern is

rare among Neotropical anurans, considering that more

pronounced declines generally occur at mid- and high

elevations; in Panama, populations declined dramatically

in some highlands, but in Costa Rica they seem to be

recovering in areas above 800 masl (Savage 2002; Pounds

et al. 2008b; Leenders 2016). A female was found inside

a rolled leaf at 100 cm above the forest floor attending

a mass of 29 eggs. This same behavior was previously

reported in Panamanian populations by Myers (1969).

Recent research showed that Craugastor persimilis (VU)

is susceptible to habitat fragmentation and it is often

absent in open pasture lands and pineapple plantations

(Bolafios et al. 2008). This observation 1s consistent with

the observations reported here, as this species was only

observed in the mature premontane forest.

Duellmanohyla uranochroa (EN) was a historically

common species across humid lowland and mountain

forests (Savage 2002). However, it has declined

precipitously since the late 1980s. By 2002, D. uranochroa

had experienced a significant decline across several

populations in Costa Rica (Duellman 2001; Savage 2002;

Leenders 2016; IUCN 2019). However, since 2007 some

populations have reappeared in Monteverde, the Matama

mountains, and Tuis de Turrialba (NatureServe and IUCN

2013), as well as in western Panama (Hertz et al. 2012).

In this monitoring effort, populations were observed

at the premontane and basal belt of the mature forest,

sometimes close to riparian environments. Generally,

males were found on the forest floor or on walking palm

roots, and up to 2 m high. Similar behaviors have been

reported in other populations of this species (Duellman

2001; Savage 2002; NatureServe and IUCN 2013).

Before this report, Ecnomiohyla veraguensis was

only known from two small Panamanian populations in

Santa Fé National Park, where it 1s highly threatened by

Amphib. Reptile Conserv.

ongoing habitat modification due to forest clearance for

agriculture and open pit mining (IUCN SSC Amphibian

Specialist Group 2019). Ecnomiohyla veraguensis 1s

differentiated here from £. miliaria, another congener

from the Caribbean foothills, based on the presence of

scalloped fleshy fringes and the absence of heel tubercles

in the former (Batista et al. 2014); from E. bailarina,

considering that E. veraguensis has a finely tuberculate

dorsum (strongly tuberculate in EF. bailarina) with

scattered minute keratin tipped tubercles on the posterior

part of the body and 6—8 widely spaced, keratinized black

spines bordering the outer side of the thumb (two clusters

of numerous, small nuptial spines in EF. bailarina; Batista

et al. 2014). The most similar species to E. veraguensis is

E. sukia,; however, the latter lacks nuptial spines in adult

males (Batista et al. 2014).

Gastrotheca cornuta (EN) is considered a rare species

in Colombia and Costa Rica, while it has declined in

Ecuador and Panama (Coloma et al. 2008; AmphibiaWeb

2009). In Costa Rica, this species is known from only

three localities in the Limon Province (Coloma et al.

2008). The first specimen was collected during 1984 in

the northwest of Nimaso peak in the Talamanca Mountain

range at 700 m asl (Solorzano et al. 1998; Savage 2002).

The other two localities were reported in Veragua, also

at basal mature forests (Solorzano et al. 1998; Salazar

2015).

Bolitoglossa alvaradoi (EN) was only observed once

in the mature forest. This endemic species has only

been reported in undisturbed areas and it is considered

endangered because its extent of occurrence is less than

5,000 km? (Bolafios et al. 2008). This salamander is a

rare species, mostly due to its secretive arboreal habitats

(Bolafios et al. 2008). In the current survey, this species

was found during the day near a small stream on a leaf at

100 cm. Some other studies reported individuals inside

bromeliads and leaf axils during the day (Wake 1987;

Savage 2002).

Agalychnis lemur (CR) occurs in Costa Rica, Panama,

and marginally in Colombia (Solis et al. 2008). It inhabits

basal and premontane humid forests and has been

historically associated with pristine areas (Duellman

2001; Savage 2002). This species has always been fairly

uncommon throughout its range; however, it was listed

as Critically Endangered because of ongoing drastic

population declines, estimated to be more than 80% over

a ten-year period (Solis et al. 2008). This survey found

three separate natural breeding populations in the study

area. One of these populations was already reported in

Costa Rica and was considered the only remnant wild

breeding population (Solis et al. 2008). Another small

population was reported in Guayacan (Kubicki 2008;

Solis et al. 2008). All other previously known Costa

Rican populations of this species have disappeared,

including those in Monteverde, San Ramon, Braulio

Carrillo, and Tapanti (Solis et al. 2008).

The main threats reported for A. /emur are habitat

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Salazar-Zuniga et al.

destruction and chytridiomycosis (Solis et al. 2008). In

one of the reported populations in this study, Whitfield

et al. (2017) found a low infection prevalence (<10%,

n = 20) of Batrachochytrium dendrobatidis (Bd) and

a low infection intensity among infected individuals.

Some studies demonstrated that highly Bd-susceptible

amphibians persist in environments hostile to Bd, even

when &d 1s still present (Puschendorf et al. 2011). The

samplings reported here never registered a sick animal.

Nonetheless, in some places where it was common to see

the species during the sampling surveys, A. /emur had

disappeared after the intensification of wood extraction

during 2013. Agalychnis lemur appears to be highly

susceptible to habitat loss, and the lack of natural

reproductive sites in the forest promotes the use of the

flooding banks or small ponds at the forest edge (JASZ,

pers. obs.), a condition that we consider makes this

species extremely vulnerable to habitat fragmentation.

This study shows that the threatened species reported

here are associated with mature forest. These species

may be sensitive to changes in their environment and

might therefore exhibit a low tolerance to human impact

(Dixo and Martins 2008; Lopes-Santos et al. 2018). The

main biodiversity threats observed while conducting

this study were: 1. Habitat destruction (legal or illegal)

due to population growth, pastures, and extraction

labors for wood and stone; 2. Monocultures of extensive

plantations (e.g., by banana and pineapple corporations

in the nearby lowlands), that also create substantial soil

erosion and use numerous agrochemicals to maintain

the crops, producing substantial amounts of pollution

residues (Castillo et al. 1997; Castillo and Ruepert 2001;

Sasa et al. 2010); 3. Illegal wildlife extraction; 4. Little

control by the responsible authorities; and 5. Low levels

of environmental education.

Conclusions

This survey shows that Veragua is a high priority

conservation area with 11.7% of its amphibian diversity

under the IUCN threatened categories, out of which

five species are cataloged as Endangered or Critically

Endangered (Table 1; IUCN 2019). In addition, this

study reports E. veraguensis in Costa Rica for the first

time and represents the only locality known for E.

bailarina (Kubicki and Salazar 2015). The diversity

analysis reveals one of the most important amphibian hot

spots in the Neotropics, with evidence of recent sightings

of several species after concerning declines (like those

of Duellmanohyla uranochroa and Agalychnis lemur),

and contrasts with the decimated diversity in several

other important locations in Costa Rica (e.g., La Selva,

Rincon de Osa, Cerro Chompipe, Monteverde, Cerro de

la Muerte, Tapanti, Volcan Cacao, Palmar Norte, and Las

Tablas) that have declined or disappeared since the late

1980s (Whitfield et al. 2007; Sasa et al. 2010; Ryan et al.

2015). Based on these findings, we suggest a long-term

Amphib. Reptile Conserv.

monitoring of the biodiversity in order to have control

over population fluctuations, and we highly recommend

natural history and behavioral studies to improve

conservation actions across this biodiversity hot spot.

According to the international conservation agreements,

as well as Costa Rica’s laws and executive decrees, the

information provided in this article should help to protect

the area from invasive activities that may negatively

affect the biodiversity or major river basins (Sasa et al.

2010).

Acknowledgements.—This research was possible thanks

to the invaluable help of the following group of researchers

and field assistants: José Brenes-Andrade, Andrés Royas-

Valle, Erick Arias, Diana Salazar, Iria Chacon, Victor

Acosta-Chaves, Adrian Garcia-Rodriguez, Marcelo

Elizondo, Rolando Ramirez, Julissa Gutiérrez-Figueroa,

Irene Ossenbach, Alejandro Quesada-Murillo, Melissa

Diaz-Morales, and Diego Salas. For photographic

material, we are thankful for the collaboration of Victor

Acosta-Chaves, Esmeralda Arévalo-Huezo, Abel Batista,

Daniel Hernandez, Andreas Hertz, and Andréi Solis. We

also appreciate the help of Tim Bray and Cesar Barrios-

Amoros as external reviewers, and Steve McCormack

and Cindy Chaves as English reviewers. It is important

to highlight the participation of Luis Angel Mejia

Gonzales (Pecas), Stanley Salazar, and Julian Solano

Salazar for their great efforts in searching for and finding

the species Gastrotheca cornuta and the three species

of Ecnomiohyla; to Johnny Hernandez and the sisters

Mariana and Isabella Jiménez for finding the species

Oedipina berlini, and to Esmeralda Arévalo-Huezo

and the zoology class (2018) of the Universidad Latina

(Costa Rica) for finding the only caecilid in the survey

(C. volcani). We are very grateful for the participation

of the local people of the communities of Las Brisas,

El Peje, and the Cabecar Ethnic Group, especially to

the indigenous leader Ruperto Lopez Camacho, who

shared his knowledge and guided us into the pristine

forest. We appreciate the logistic support provided by

the School of Biology and the Museum of Zoology of

the University of Costa Rica, as well as the great help

from the employees of Veragua Rainforest Research and

Adventure Park to conduct this project. Finally, we thank

The Veragua Foundation for the Rainforest Research and

its president, José Marti Jiménez-Figueres, for financing,

believing, and supporting this important research for the

conservancy.

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José Andrés Salazar-Zufiiga is a biologist and M.Sc. student in the Department of Ecology at

Universidad Autonoma de Madrid and Universidad Complutense de Madrid, Spain. He is a

professor of herpetology and an active researcher of the Herpetology Department of the University

of Costa Rica, and research coordinator of the Veragua Foundation, Limon, Costa Rica (NGO).

José’s research interests include animal behavior, natural history, and conservation of amphibians

and reptiles. He has participated in several conservation workshops organized by the Amphibian

a > Specialist Group and the University of Costa Rica, including the workshops to review the IUCN

a *% . | | ii \

Red List of amphibians of Costa Rica and for elaborating the amphibian conservation strategy for

Mesoamerica. José is an active conservationist who participates in different environmental education

programs in rural communities, and he is currently developing different research and conservation

projects with various species of the Centrolenidae, Dendrobatidae, and Hylidae families.

Wagner Chaves-Acufia is a Ph.D. student in the Department of Biodiversity and Experimental

Biology at Universidad de Buenos Aires, Argentina, and a fellow of Consejo Nacional de

Investigaciones Cientificas y Técnicas (CONICET) at the Museo Argentino de Ciencias Naturales

“Bernardino Rivadavia,’” Argentina (MACN). Wagner is also an associated researcher at Veragua

Foundation, where he has conducted research on bioacoustics and behavior of dendrobatids and

centrolenids. His current research interests include systematics, taxonomy, and evolution of hylids,

as well as conservation projects with critically endangered species of anurans.

Gerardo Chaves is a biologist from the University of Costa Rica. Gerardo’s degree thesis focused

on the arrivals of the Olive Ridley Sea Turtles, but most of his professional work has focused on

the ecology and taxonomy of the Costa Rican herpetofauna. Since 1992, his research activity has

focused on understanding the decline of amphibian populations in Mesoamerica and on filling the

herpetofauna inventory gaps in several areas of Costa Rica, mainly across the Talamanca Mountain

Range. Since 1997, Gerardo has worked in the Museum of Zoology of the University of Costa Rica

in the herpetofauna collection. He has published several journal articles related to the ecology and

taxonomy of Neotropical herpetofauna. His conservation efforts are related to the sustainable use of

the sea turtle eggs project on “arribadas” and collaboration with IUCN in the evaluation of the Red

List for Costa Rica and Mesoamerica, for both reptiles and amphibians. Gerardo is currently chair

of the IUCN Amphibian Specialist Group in Costa Rica.

Amphib. Reptile Conserv. 224 December 2019 | Volume 13 | Number 2 | e215

Amphib. Reptile Conserv.

Extreme frog diversity in Costa Rica

Alejandro Acufia is a professional in Ecotourism Management, and he has worked as a coordinator

of biodiversity projects in several national parks with the National System of Conservancy Areas

(SINAC). Alejandro is a naturalist guide at Veragua Rainforest and research assistant of the Veragua

Foundation.

Juan Ignacio Abarca-Odio is a biologist from the University of Costa Rica, where he collaborates

as a researcher in the Aquatic Experimentation Laboratory (CIMAR) and the Laboratory for

Experimental and Comparative Pathology (LAPECOM). Juan’s main interests are on the effects

of climate change on the ecology and physiology of vulnerable organisms such as anthozoans,

arthropods, amphibians, and reptiles. He also has great interest in data science.

Javier Lobon-Rovira is a Wildlife Photographer and a Ph.D. student at the CIBIO-InBio institution

in Portugal. Javier has assisted as an Animal Care Volunteer at Wildlife Rescue Association

(Vancouver, British Columbia, Canada) rehabilitating wildlife and promoting the welfare of wild

animals in the urban environment. He has worked as a field assistant with Moose and Wolves in

Utah, and sampling fishes using electric-fishing techniques. For his Master's thesis, Javier identified

a “lost population” of Iberian Lynx by anecdotal occurrence data and molecular scatology, which

formed a major part of his M.Sc. degree. Furthermore, he has collaborated on many different

herpetology and conservation projects. Currently, his Ph.D. project focuses on the systematics and

evolution of geckonids from Southern Africa, which includes the descriptions of species and the

identification of several evolutionary hypotheses within this group.

Edwin Gémez-Méndez is biologist from the University of Costa Rica. Edwin has work as an

Environmental Manager in several important national projects, such as the Reventazon Hydroelectric

Project, expansion of the National Route 32, and the Costa Rica-Panama Binational Bridge. Edwin

is a Biology professor at Florencio del Castillo University, Costa Rica. His research interests concern

the conservation of amphibians and reptiles.

Ana Cecilia Gutiérrez-Vannucchi is a biologist and M.Sc. student at the School of Biology of

the University of Costa Rica, where she is part of the Laboratory of Urban Ecology and Animal

Communication (LEUCA). Ana’s research interests are in ecology, bioacoustics, and animal

behavior. Her most recent projects have focused on studying the possible effects of urban noise on

the acoustic communication of anurans.

Federico Bolaiios is a professor of Herpetology at the School of Biology of the University of Costa

Rica, curator of the Herpetology collection at Museo de Zoologia, and a member of the International

Union for Conservation and Nature (Amphibian, Conservation Breeding, and Viper Specialist

Groups). Federico’s M.Sc. dissertation focused on the natural history and population ecology of

Oophaga granulifera. His primary interest involves the behavioral ecology of amphibians, but he

has also participated in taxonomic studies, including the description of nine species of amphibians.

Federico became a professor when amphibian declines were first being detected and has since

dedicated most of his research efforts to this topic. He has mentored more than 35 graduate students

in Biology at UCR. He has authored more than 70 publications, including book chapters and peer

reviewed papers in scientific journals, and has served as a book editor.

322 December 2019 | Volume 13 | Number 2 | e215

Official journal website:

amphibian-reptile-conservation.org

Amphibian & Reptile Conservation

13(2) [General Section]: 323-354 (e216).

urn:lsid:zoobank.org:pub:E0578CD4-6843-42A9-9E23-25B8C23FC1DA

Three new species of day geckos (Reptilia: Gekkonidae:

Cnemaspis Strauch, 1887) from isolated granite cave

habitats in Sri Lanka

1*Suranjan Karunarathna, 7Anslem de Silva, **Madhava Botejue, *Dinesh Gabadage, ®Lankani

Somaratna, ‘*Angelo Hettige, ‘Nimantha Aberathna, °®Majintha Madawala, **Gayan Edirisinghe,

‘Nirmala Perera, ‘*Sulakshana Wickramaarachchi, °Thilina Surasinghe, '*Niranjan Karunarathna,

‘°Mlendis Wickramasinghe, ‘Kanishka D.B. Ukuwela, and ‘*Aaron M. Bauer

'Nature Explorations and Education Team (NEET), No: B-1 / G-6, De Soysapura Flats, Moratuwa 10400, SRI LANKA Amphibia and Reptile

Research Organization of Sri Lanka (ARROS), 15/1, Dolosbage Road, Gampola 20500, SRI LANKA °Biodiversity Conservation Society (BCS),

150/6, Stanly Thilakaratne Mawatha, Nugegoda 10250, SRI LANKA *Central Environmental Authority (CEA), 104, Denzil Kobbekaduwa Mawatha,

Battaramulla 10120, SRI LANKA °Zoology Division, Department of National Museums, Colombo 07, SRI LANKA °Young Zoologist Association of

Sri Lanka (YZA), Department of National Zological Gardens, Dehiwala 10350, SRI LANKA ‘Youth Exploration Society of Sri Lanka (YES), Royal

Botanical Garden, Peradeniya 20400, SRI LANKA 8Victorian Herpetological Society (VHS), P.O. box 4208, Ringwood, VIC 3134, AUSTRALIA

°Department of Biological Sciences, Bridgewater State University, Bridgewater, MA 02325, USA '°Herpetological Foundation of Sri Lanka (HFS),

31/5, Alwis Town, Hendala, Wattala 11300, SRI LANKA ''Department of Biological Sciences, Faculty of Applied Sciences, Rajarata University of

Sri Lanka, Mihintale 50300, SRI LANKA '*Department of Biology, Center for Biodiversity and Ecosystem Stewardship, Villanova University, 800

Lancaster Avenue, Villanova, Pennsylvania 19085, USA

Abstract.—Three new day gecko species of the genus Cnemaspis Strauch, 1887 are described from three

isolated granite cave habitats with rock walls in Bambaragala (Ratnapura District), Dimbulagala (Polonnaruwa

District), and Mandaramnuwara (Nuwara-Eliya District) in Sri Lanka based on morphometric and meristic

characters. All of these new species are assigned to the kandiana clade based on morphology. These species

are small (28-35 mm SVL) in size and may be differentiated from all other Sri Lankan congeners by a suite

of distinct morphometric and meristic characters. Each of these species described herein are categorized

as Critically Endangered (CR) under IUCN Red List criteria. At the microhabitat scale, they are restricted to

wet, cool, and shady granite caves and rock outcrops in isolated forested areas with limited anthropogenic

disturbance. Further, these habitats are located in all three main bioclimatic zones (wet, intermediate, dry)

and all three geographic peneplains (first, second, third) of Sri Lanka. Due to their restricted distributions

(as point endemics), the habitats of these specialist species are vulnerable to fragmentation, edge effects,

and anthropogenic activities. Therefore, these isolated forest patches in Sri Lanka are in need of special

conservation attention and management.

Keywords. Climate condition, endangered species, habitat specialist, isolated forest, point endemic, range restriction,

systematics, taxonomy

Citation: Karunarathna S, de Silva A, Botejue M, Gabadage D, Somaratna L, Hettige A, Aberathna N, Madawala M, Edirisinghe G, Perera N,

Wickramaarachchi S, Surasinghe T, Karunarathna N, Wickramasinghe M, Ukuwela KDB, Bauer AM. 2019. Three new species of day geckos (Reptilia:

Gekkonidae: Cnemaspis Strauch, 1887) from isolated granite cave habitats in Sri Lanka. Amphibian & Reptile Conservation 13(2) [General Section]:

323-354 (e216).

tribution 4.0 International (CC BY 4.0): https://creativecommons.org/licenses/by/4.0/], which permits unrestricted use, distribution, and reproduction in

any medium, provided the original author and source are credited. The official and authorized publication credit sources, which will be duly enforced,

are as follows: official journal title Amphibian & Reptile Conservation; official journal website: amphibian-reptile-conservation.org.

Received: 28 May 2019; Accepted: 4 December 2019; Published: 31 December 2019

Introduction sequently, Cnemaspis ranks as the second most diverse

gecko genus in the world, next to Cyrtodactylus (Gris-

Taxonomic descriptions and phylogenetic revisions in

the past decade have rapidly increased the number of

day gecko species recognized in the genus Cnemaspis,

bringing the global species richness to more than 155

(Karunarathna et al. 2019a,b; Uetz et al. 2019a). Con-

Correspondence. *suranjan.karu@gmail.com

Amphib. Reptile Conserv.

mer et al. 2014; Uetz et al. 2019a). However, extensive

molecular phylogenetic analyses have questioned the

monophyly of Cnemaspis which is represented by three

geographically disjunct groups from South Asia, Tropi-

cal Africa, and Southeast Asia (Gamble et al. 2012; Py-

December 2019 | Volume 13 | Number 2 | e216

Three new species of Cnemaspis from Sri Lanka

ron et al. 2013a; Zheng and Wiens 2016). Cnemaspis

geckos are diminutive, slender-bodied geckos that pos-

sess prominent forward and upwardly-directed eyes with

round pupils, broad flattened heads, and elongate slender

digits that are bent at an angle with entire subdigital la-

mellae (Vidanapathirana et al. 2014; Wood et al. 2017;

Karunarathna et al. 2019a). These geckos are adapted for

a scansorial mode of life, with most being rupicolous,

while a few are arboreal or ground-dwelling with crepus-

cular behavior (Das 2005). They appear to be microhabi-

tat specialists with occupancy limited to shaded surfaces

of rocks, caves, trees, abandoned buildings, buildings as-

sociated with caves, wattle and daub houses, and rock

walls within suitable habitats where the cryptic morphol-

ogy and body coloration help them camouflage with their

surroundings (Smith 1935; Karunarathna et al. 2019b).

Much like continental south Asia, as well as the Indo-

Malayan realm, the species richness of Cnemaspis in Sri

Lanka has grown rapidly by at least eight-fold, from four

to 33 species (Deraniyagala 1953; Manamendra-Arach-

chi et al. 2007; Wicramasinghe and Munindradasa 2007;

Karunaratha and Ukuwela 2019). As such, Cnemaspis

has become the most diverse gecko genus on the island,

with 100% endemism. Through molecular phylogenetic

analyses of mitochondrial and nuclear DNA, Agarwal et

al. (2017) demonstrated the presence of two distinct Cne-

maspis Clades in Sri Lanka (kandiana and podihuna), and

indicated the presence of cryptic diversity within four

species (C. alwisi Wickramasinghe and Munidradasa

2007; C. kumarasinghei Wickramasinghe and Munidra-

dasa 2007; C. latha Manamendra-Arachchi, Batuwita,

and Pethiyagoda 2007, and C. podihuna Deraniyagala

1944). The aforementioned studies emphasized the need

for detailed studies on Cnemaspis taxonomy using a

combination of both morphological characteristics and

molecular phylogenetics. As indicated by recent stud-

ies in Sri Lanka, the faunistic surveys of under-explored

habitats followed by rigorous phylogenetic analyses will

further augment the species richness of Cnemaspis (Bau-

er et al. 2007; Agarwal et al. 2017; Karunarathna et al.

2019b). In light of this, we conducted field excursions in

various isolated localities in Sri Lanka. Here, we describe

three new Cnemaspis species (based on morphometric

and meristic characters) discovered from three such sites

which span all three bioclimatic zones and geological pe-

neplains of Sri Lanka.

Materials and Methods

Field sampling and specimens. Field surveys across 122

different locations in Sri Lanka covered several geograph-

ic areas (e.g., dry zone, intermediate zone, and wet zone).

At each location, gecko species found were surveyed and

documented with special attention on the focal genus. On

average, 12 surveyor-hours per location were devoted to

the survey. Museum acronyms follow Sabaj (2016) and

Uetz et al. (2019b). The type materials discussed in this

Amphib. Reptile Conserv.

paper are deposited in the National Museum of Sri Lanka

(NMSL), Colombo, Sri Lanka. Specimens were caught

by hand and were photographed in life. They were eutha-

nized using halothane and fixed in 10% formaldehyde for

two days, washed in water, and then transferred to 70%

ethanol for long-term storage. Tail tips were collected as

tissue samples before fixation and were stored in 95%

ethanol under relatively cool conditions (20—25 °C). For

comparison, 424 Cnemaspis specimens (catalogued and

uncatalogued) representing all recognized Sri Lankan

species were examined, including all type specimens

housed at the National Museum, Sri Lanka (NMSL), The

Natural History Museum, London (BMNH), and in the

private collections of Anslem de Silva (ADS) and Aaron

Bauer (AMB), which had been deposited in the NMSL.

Specimens that formerly belonged to the Wildlife Heri-

tage Trust (WHT) collection and bear WHT numbers are

currently deposited in the NMSL, catalogued under their

original numbers. Specimens in this study were collected

during a survey of lizards in Sri Lanka under permit num-

bers WL/3/2/1/14/12 and WL/3/2/42/18 (a and b), issued

by the Department of Wildlife Conservation, and under

permit numbers FRC/5 and FRC/6, issued by the Forest

Department of Sri Lanka. Additional information on the

morphology and natural history of Sri Lankan Cnemaspis

species was extracted from the relevant literature (Bauer

et al. 2007; Manamendra-Arachchi et al. 2007; Wick-

ramasinghe and Munindradasa 2007; Vidanapathirana

et al. 2014; Wickramasinghe et al. 2016; Batuwita and

Udugampala 2017; Agarwal et al. 2017; Batuwita et al.

2019; Karunarathna et al. 2019a,b; de Silva et al. 2019).

Assignment of unidentified specimens to the three new

Species was based on their morphometric and meristic

characters (Tables 1-9), color patterns, and geographic

isolation (Fig. 1; Table 10). The new species described in

the present paper are completely new and have not been

included in previous phylogenies of the genus (see A gar-

wal et al. 2017; Karunarathna et al. 2019b).

Morphometric characters. Forty morphometric mea-

surements were taken using a Mitutoyo digital Vernier

calliper (to the nearest 0.1 mm), and detailed observa-

tions of scales and other structures were made through

Leica Wild M3Z and Leica EZ4 dissecting microscopes.

The following symmetrical morphometric characters

were taken on the left side of the body: eye diameter

(ED), horizontal diameter of eye ball; orbital diameter

(OD), greatest diameter of orbit; eye to nostril length

(EN), distance between anteriormost point of orbit and

posterior border of nostril; snout length (ES), distance

between anteriormost point of orbit and tip of snout;

snout to nostril length (SN), distance between tip of

snout and anteriormost point of nostril; nostril width

(NW), maximum horizontal width of nostrils; eye to ear

distance (EE), distance between posterior border of eye

and anteriormost point of ear opening; snout to axilla

distance (SA), distance between axilla and tip of snout;

December 2019 | Volume 13 | Number 2 | e216

Karunarathna et al.

Contour 600 m

Climate zone boundary

Dry zone

_ Intermediate zone

, Wet zone

;

L " Kilometers

WeGe 1984 UTM Zone 44h

Fig. 1. Currently known distribution of Cnemaspis dissanay-

akai sp. nov. (square) from Dimbulagala; Cnemaspis kawmin-

iae sp. nov. (circle) from Mandaramnuwara; and Cnemaspis

kotagamai sp. nov. (triangle) from Bambaragala, Sri Lanka.

ear length (EL), maximum length of ear opening; interor-

bital width (IO), shortest distance between left and right

supraciliary scale rows; inter-ear distance (IE), distance

across head between the two ear openings; head length

(HL), distance between posterior edge of mandible and

tip of snout; head width (HW), maximum width of head

between the ears and the orbits; head depth (HD), maxi-

mum height of head at the level of the eye; jaw length

(JL), distance between tip of snout and corner of mouth;

internarial distance (IN), smallest distance between in-

ner margins of nostrils; snout to ear distance (SED), dis-

tance between tip of snout and anteriormost point of ear;

upper-arm length (UAL), distance between axilla and

the angle of the elbow; lower-arm length (LAL), dis-

tance from elbow to wrist with palm flexed; palm length

(PAL), distance between wrist (carpus) and tip of longest

finger excluding the claw; length of digits I-V of manus

(DLM), distance between juncture of the basal phalanx

with the adjacent digit and the tip of the digit, excluding

the claw; snout-vent length (SVL) distance between tip

of snout and anterior margin of vent; trunk length (TRL),

distance between axilla and groin; trunk width (TW),

Amphib. Reptile Conserv.

maximum width of body; trunk depth (TD), maximum

depth of body; femur length (FEL), distance between

groin and knee; tibia length (TBL), distance from knee to

heel with ankle dorsiflexed; heel length (HEL), distance

between ankle (tarsus) and tip of longest toe (excluding

the claw) with both foot and tibia flexed; length of pedal

digits I-V (DLP), distance between juncture of the basal

phalanx with the adjacent digit and the digit tip, exclud-

ing the claw; tail length (TAL), distance between anterior

margin of the vent and tail tip; tail base depth (TBD),

maximum height of tail base; tail base width (TBW),

widest point of tail base.

Meristic characters. Thirty discrete characters were ob-

served and recorded using Leica Wild M3Z and Leica

EZ4 dissecting microscopes on both the left (L) and the

right (R) sides of the body (reported in the form L/R):

number of supralabials (SUP) and infralabials (INF) be-

tween the first labial scale and corner of mouth; number of

interorbital scales (INOS), between left and right supra-

ciliary scale rows; number of postmentals (PM) bounded

by chin scales, 1“ infralabial on the left and right and

the mental; number of chin scales (CHS), scales touching

medial edge of infralabials and mental between juncture

of 1‘ and 2" infralabials on the left and right; number of

supranasal (SUN) scales between nares; presence of the

postnasal (PON) scales posterior to naris; presence of the

internasal (INT) scale between supranasals; number of

supraciliary scales (SUS) above eye; number of scales

between eye and tympanum (BET) from posteriormost

point of orbit to anteriormost point of tympanum; num-

ber of canthal scales (CAS), number of scales from pos-

teriormost point of naris to anteriormost point of orbit;

total lamellae on manus I-V (SLM) counted from first

proximal enlarged scansor greater than twice width of the

largest palm scale, to distalmost lamella at tip of digits;

number of dorsal paravertebral granules (PG) between

pelvic and pectoral limb insertion points along a straight

line immediately left of vertebral column; number of

midbody scales (MBS) from the center of mid-dorsal

row diagonally towards the ventral scales; number of

midventral scales (MVS) from the first scale posterior to

the mental to last scale anterior to vent; number of belly

scales (BLS) across venter between the lowest rows of

granular dorsal scales; total lamellae on pes I-V (SLP),

counted from first proximal enlarged scansor greater than

twice the width of the largest heel scale, to distalmost

lamella at tip of digits; number of precloacal pores (PCP)

anterior to the cloaca; number of femoral pores (FP)

present on femur; numbers of non-pored proximal femo-

ral scales (PFS) counted from proximal ends of femo-

ral pore rows to precloacal pores; numbers of non-pored

distal femoral scales (DFS) counted from distal ends of

femoral pore rows to knee; interfemoral scales (IFS)

number of non-pored scales between femoral pores on

both femurs. Additional evaluations included the texture

[keeled (KD) or smooth (SM)] of the ventral scales, the

December 2019 | Volume 13 | Number 2 | e216

Three new species of Cnemaspis from Sri Lanka

texture [heterogeneous (HET) or homogeneous (HOM)|

of the dorsal scales, the number of spinous scales on the

flanks (FLSP), and characteristics such as appearance of

the caudal scales (except in specimens with regenerated

tails). Coloration was determined from digital images of

living specimens and also from direct observations in the

field.

Distribution and natural history. The new species de-

scribed herein were collected during field surveys con-

ducted in various habitats (e.g., dry mixed semi-evergreen

forest and tropical wet-evergreen) of Sri Lanka (Fig. 1;

Table 10). During these surveys, behavioral and other

aspects of natural history of the focal species were ob-

served through opportunistic and non-systematic means.

The ambient and substrate temperatures were measured

using a standard thermometer and an N19 Q1370 infrared

thermometer (Dick Smith Electronics, Shanghai, China),

respectively. The relative humidity and light intensity

were measured with a QM 1594 multifunction environ-

ment meter (Digitek Instruments Co., Ltd., Hong Kong,

China). To record elevation and georeference species lo-

cations, an eTrex® 10 GPS (Garmin) was used. Sex was

determined by the presence in males (M) or absence in

female (F) of hemipenal bulges, and precloacal and fem-

oral pores. The conservation status of each new species

was evaluated using the 2001 IUCN Red List Categories

and Criteria version 3.1 (IUCN 2012).

Statistical analyses of morphometric characteristics.

Principal Component Analyses (PCA) were performed

with the conventional singular value decomposition

method using variance-covariance stricture as the cross-

products matrix to extract 10 principal components

(package: PCAMethods, function:pca; Stacklies et al.

2007). The species used for these analyses are: Cnemas-

pis dissanayakai sp. nov., Cnemaspis ingerorum, Cne-

maspis kallima, Cnemaspis kawminiae sp. nov., Cne-

maspis kotagamai sp. nov., Cnemaspis kumarasinghei,

Cnemaspis latha, and Cnemaspis gotaimbarai groups

due to their close resemblances. All morphometric mea-

surements of the three new species were normalized

to the snout-vent length (SVL). The matrix containing

Table 1. Principal component scores and corresponding species.

original morphometric variables were Pareto scaled

(square-root unit variance) and centered. Subsequently,

pairwise ordination plots were generated for the first

four principal components (PC), which explained nearly

80% of the cumulative variance, where each individual

PC accounted for at least 8% of the overall variance. To

visualize species separation in the ordination space, con-

vex hulls were placed around PC scores corresponding to

each species (Wickham 2016). In addition, PC loadings

were examined against each of the selected PC axes to

understand the relationships between the original mor-

phometric variables and the PC axes. This examination

also revealed which morphometric variables were most

distinct among the different species. In addition to the

collective analyses of the eight aforementioned conge-

nerics, three separate PCAs were run which focused on

three species groups based on their close morphological

resemblances: (1) Cnemaspis kotagamai sp. nov., Cne-

maspis ingerorum, and Cnemaspis kallima; (2) Cnemas-

pis dissanayakai sp. nov., Cnemaspis latha, and Cne-

maspis kumarasinghei;, and (3) Cnemaspis kawminiae

sp. nov., Cnemaspis gotaimbarai, and Cnemaspis kuma-

rasinghei. The aforementioned analyses used statistical

program R (R Core Team 2019) and RStudio integrated

development environment (R Studio Team 2018). Ordi-

nation plots were produced using the following statisti-

cal applications and R packages: PAST version 3.14 and

geplot2 (Hammer et al. 2001).

Results

Analyses of morphometric data for all eight species.

The PCA resulted in 10 PCs that accounted for 98% of

the variability of the original morphometric variables;

among these, PC 1-4 (35.64%, 19.48%, 14.35%, and

8.70%, respectively) cumulatively explained 78% of the

overall variability (Fig. 2). Trunk and upper-arm lengths

had greater loadings on PC1 while tail length, orbital di-

ameter, snout-to-nostril length, and the lengths of the 4"

and 5" pestal digits had greater loadings on PC2. Heel

length, snout-to-axilla length, eye diameter, inner-ear dis-

tance, eye-to-nostril distance, trunk width, and length of

the 4" finger had higher loadings on PC3; whereas trunk

Average scores for principal components

Species

PCl1 PC2

C. kotagamai 0.31 0.52

C. dissanayakai -0.50 0.20

C. kawminiae 0.40 -0.05

C. kumarasinghei 0.12 -0.28

C. gotaimbarai 0.44 -0.47

C. ingerorum 0.15 0.24

C. latha -0.60 -0.03

C. kallima -0.33 -0.13

Amphib. Reptile Conserv.

326

PC3 PC4 PCS PC6

0.31 0.12 0.16 0.00

0.17 -0.22 0.01 -0.10

-0.23 -0.27 0.13 0.34

0.09 -0.02 0.25 -0.21

0.20 0.06 -0.20 -0.06

-0.63 0.10 -0.09 -0.19

0.025 0.37 -0.03 0.17

0.07 -0.14 -0.23 0.03

December 2019 | Volume 13 | Number 2 | e216

Karunarathna et al.

A C B oO

3 & 0.25-

& 05 7

5 a : is

> = 0.00-

c) ok, 6 f

= a =

4 O.0- t RE 4 mal a 0.25"

a oO Zz

ow on “0.50

oO oO

a -05 a

“0 5 4 i ‘ ‘

“1.0 “0.5 0.0 05 1.0 “0.5 0.0 os

PC1 35.64 % of vanance PC1 35.64 % of vanance

x rr)

i AS D 7

oily o = ‘ =

a 35 - =

Ee WP _ — 5 al 3 oF es)

oS ; oO C oa CS

a sa en a ee se

oo -D.2 =

s f = A

3 “0.25 on 050°

a : a

-0.50- : he ; ; “0.75: ; : Mv

1.0 “0.5 00 0.5 0.5 0.0 05

PC1 35.64 % of variance PG2 19.48% of vanance

E ona F He

om ow

e 0.25 i & 0.25- 4

5 . xi Gl oO Fa - Wy r 4 a)

peal ST = 7 a, = —4">

2 0.00- ‘ * -L + : 2 0.00- + <i

[ri] Lea] a a

Lia] Lewy

oo o

“0.25 - “0.25 -

tT TT

O Oo

a a

0.50" — 0.50 , ; _ & :

“0.5 0.0 05 0.75 0.50 0.25 0.00 0.25

PC2 19.48 % of variance PC3 14.35 % of variance

species |) kotagamai |) dissanayakai a kawminiae “BS kumarasinghei ) gotaimbarai GF ingerorum | latha ) kallima

. H 084 or |

sie 0.08-+ soa | o

! [ *, | 9.0504

- ~ Oe ce ee ee se i = ee | = L e !

3” ; z |. =. =2 = -_| 20,0254 * |

= = Li eS na Sie | 2 ' i

‘i = a | @ ee ee 2 ew as eae |

cL ‘a | | [as 1 !

= 0.00 & 0.00- r-- | # 0.000-+ |

m9 Ho bees £2 cee Eee mcpuae sea

line! te] Go } H " aa! Se }

~ 0.04 Romatick ec cada oe Sec ees es | =0.0255 |

2 dae Ribs na ade Sack 12 | es |

| |

b- ----- rh ----- <b ------r------ bh.

0.08 Pen sukioe ela oe Sepia Ee] |

D084 --- ---p 2-2-2 52-22 ep a-a- ee | i }

; [Pe ntl A ee Sewn Oe Ne ered ee oat a | | ry |

“0.16 “0.08 0.00 0.08 O16 0.16 -0.08 0.00 0.08 0.16 “0.16 -0.08 0.00 0.08 0.16

PC1 (49.3% explained var.) PC1 (42.5% explained var.) PC1 (64.5% explained var.)

Fig. 2. (A-F) PCA Ordination plots for multivariate morphometric analyses (all pairwise ordination plots for PC1 through PC4

are shown), (G) Scatter plot of PCA between Cnemaspis kotagamai sp. nov. (filled circles), Cnemaspis ingerorum (triangles),

and Cnemaspis kallima (filled diamonds), (H) Scatter plot of PCA between Cnemaspis dissanayakai sp. nov. (circles), Cnemas-

pis latha (filled squares), and Cnemaspis kumarasinghei (stars), (I) Scatter plot of PCA between Cnemaspis kawminiae sp. nov.

(squares), Cnemaspis gotaimbarai (filled triangles), and Cnemaspis kumarasinghei (stars).

and tail lengths had higher loadings on PC4 (Tables 1—2).

The ordination for PC1 and PC2 provided the strongest

evidence for morphometric-based species separation—

expect for Cnemaspis kawminiae, C. kumarasinghei, C.

kallima, and C. latha, whose separation was most evident

in ordination between PC1 and PC4 (Fig. 2). In addition,

PC1 and PC3 supported separation of species based on

morphometrics relatively well.

Amphib. Reptile Conserv.

327

Analyses of morphometric data for separate species

groups. PCA of morphometric measurements of Cne-

maspis kotagamai sp. nov., Cnemaspis ingerorum, and

Cnemaspis kallima indicated the presence of three well

separated species (Fig. 2). Cnemaspis kotagamai sp. nov.

and Cnemaspis kallima were clearly separated from the

PC2 axis, while Cnemaspis ingerorum and Cnemaspis

kallima were seprated along PC1. The PC1 and PC2 axes

December 2019 | Volume 13 | Number 2 | e216

Three new species of Cnemaspis from Sri Lanka

Table 2. Morphometric variables and corresponding PC load- Table 3. Morphometric data of holotype and two paratypes of

ings. Cnemaspis kotagamai sp. nov. from Bambaragala, Ratnapura

Measurements PCL PC2 PC3 PCA District, Sri Lanka.

ED -0.14 0.02 0.15 0.00 NMSL NMSL NMSL

2019.15.01 2019.15.02 2019.15.03

OD -0.22 0.21 0.06 -0.03 Measurenents —_—_—_———————————————————

Holotype Paratype Paratype

EN 0.01 -0.14 0.16 -0.04 (Male) (Male) (Female)

ES -0.08 -0.05 -0.12 -0.19 SVL 29 8 31.1 39-6

SN -0.13 0.18 0.03 0.06 TRL 12.6 123 12.5

NW 0.02 0.04 -0.07 — -0.04 TW 5.4 53 54

EE 0.10 -0.05 0.03 -0.10 TD 3.4 3.4 3.4

SA 0.15 -0.40 0.18 -0.15 TAL 33.5 33.8 31.1

EL -0.08 0.14 G20 SEIS TBW 3.3 31 2.9

IO -0.22 0.05 0.07 0.06 TBD 2.9 2.9 2.7

IE 0.20 0.02 0.15 -0.03 ED 1.9 1.9 1.8

HL 0.18 -018 0.05 -0.27 OD 3.2 3.3 3.1

HW 016 “011 <013» 0.04 EN 2.8 2.8 27

HD 0.08 -014 -0.24 -0.07 ES 3.6 3.5 ay

JL 0.00, SO.108 <0:17 0.06 on me a LS

NW 0.2 03 0.2

IN -0.16 0.11 0.05 0.03

EE 2.5 2.5 23

SED -0.23 0.08 -0.01 0.12

SA 12.9 12.7 11.8

UAL O29 -=6:07 0.03 0.15 me es a ws

LAL O15 -0.26 -0.03 0.18 in se Ae ae

PAL -0.04 0.07 -0.28 -0.13 - 38 69 38

DLM -1 -0.06 -0.02 -0.03 -0,27 HL 83 83 82

DLM-2 -0.01 — -0.06 0.00 -0.22 Hw 45 Aa 45

DLM-3 -0.10 -0.03 0.08 -0.17 HD 28 26 OF

DLM-4 -0.15 -0.06 0.14 -0.18 JIL 49 48 49

DLM-5 -0.17 0.02 0.12 -0.06 IN 1.6 1.6 1.4

TRL 0.22 -0O.13 -0.25 -0.48 SED 8.7 8.6 8.7

TW -0.22 0.11 0.14 -0.15 UAL 3.8 34) 39

TD -0.16 0.11 0.10 -0.09 LAL 3.4 3:3 Ee

FEL -0.16 -0.07 0.05 -0.13 PAL 3.2 3.2 3.2

TBL 1020 «003 ST DLM (i) 1.4 1.4 1.3

HEL 0.01 0.06 -048 0.22 DLM (it) 1.9 18 18

DLP-1 0.04 007 -031 -0.14 oe 2.5 25 2.6

DLP-2 0.24 013 -020 -0.10 DEM) ay uh

DLM (vy) phe 23 23

DLP-3 O19 -0.03 -0.15 -0O11

FEL 5.8 5.8 5.7

DLP-4 -0.19 0.16 -0.24 0.04

TBL 52 5.1 49

DLP-5 4) 3 0.21 -0.20 -0.02

HEL 48 45 47

TAL O15 -0.62 -0.20 0.24 DLP (i) a i Vs

TBW -0.14 -0.03 -0.01 0.22 DLP (ii) ae ro a4

TBD -O.11 0.05 0.01 0.21 DLP (iii) 9 28 07

DLP (iv) 3.8 3.9 3.7

DLP (v) 3.5 33 3.5

Amphib. Reptile Conserv. 328 December 2019 | Volume 13 | Number 2 | e216

Karunarathna et al.

= i

: ;

fh te

Ri saaras r >

ee ee es eg

ae Seah: a

i, eel

Fig. 3. Holotype male of Cnemaspis kotagamai sp. nov. (NMSL 2018.15.01). (A) Dorsal head, (B) lateral head, (C) ventral head,

(D) heterogeneous dorsal scales, (E) scales on lateral surface of trunk, (F) smooth ventral scales, (G) cloacal characters with pre-

cloacal pores and femoral pores, (H) lamellae on manus, (I) lamellae on pes, (J) keeled dorsal scalation of tail, (IX) lateral side of

tail, and (L) very small smooth subcaudals. Photos: Suranjan Karunarathna.

explained 49.5% and 27.3% of the observed variation,

respectively. Analysis of morphometric measurements of

Cnemaspis dissanayakai sp. nov., Chemaspis latha, and

Cnemaspis kumarasinghei similarly indicated the pres-

ence of three well separated species (Fig. 2). Cnemas-

pis dissanayakai sp. nov. and Cnemaspis kumarasinghei

Amphib. Reptile Conserv.

were well separated in the PC1 axis while the former was

clearly separated from Cnemaspis latha in the PC2 axis.

The PC1 and PC2 axes explained 42.5% and 32.5% of

the observed variation, respectively. PCA analysis of the

morphometric measurements of Cnemaspis kawminiae

sp. nov., Cnemaspis gotaimbarai, and Cnemaspis kuma-

December 2019 | Volume 13 | Number 2 | e216

Three new species of Cnemaspis from Sri Lanka

— ee *.

® a.

Se.

uw oo [Pa

Fig. 4. Holotype male of Cnemaspis kotagamai sp. nov. (NMSL 2018.07.01) in life in-situ in Bambaragala. (A) Dorsal view of the

full body, and (B) ventral view with scattered yellow coloration. Photos: Suranjan Karunarathna.

rasinghei indicated the presence of three well separated

species (Fig. 2). Cnemaspis kawminiae sp. nov. was

clearly separated from Cnemaspis gotaimbarai along

the PC2 axis, while it was distinguished from C. kuma-

rasinghei also in the PC2 axis. The PCI and PC2 axes

explained 64.5% and 17.1% of the observed variation,

respectively.

Systematics

Cnemaspis kotagamai sp. nov. Karunarathna, de Sil-

va, Boteyue, Surasinghe, Wickramasinghe, Ukuwela &

Bauer

Kotagama’s Day Gecko (English)

Kotagamage Diva-seri Hoona (Sinhala)

Kotagamavin Pahalpalli (Tamil)

Figs. 3-5; Tables 3-4

urn:lsid:zoobank.org:act:35801F43-7148-4590-BE38-4214C8905646

Holotype. NMSL 2019.15.01, adult male, 29.8 mm SVL

(Fig. 3), collected from a granite cave Bambaragala, Pal-

Amphib. Reptile Conserv.

lebedda, Ratnapura District, Sabaragamu Province, Sri

Lanka (6.512978°N, 80.750306°E, WGS1984; elevation

127 m; around 1100 hrs) on 18 January 2019 by Suranjan

Karunarathna and Anslem de Silva.

Paratypes. NMSL 2019.15.02, adult male, 31.1 mm

SVL, and NMSL 2019.15.03, adult female, 32.6 mm

SVL, collected from a granite cave in Bambaragala,

Pallebedda, Ratnapura District, Sabaragamuwa Prov-

ince, Sri Lanka (6.517261°N, 80.752692°E, WGS1984;

elevation 132 m; around 1200 hrs) on 18 January 2019

by Suranjan Karunarathna and Anslem de Silva.

Diagnosis. Cnemaspis kotagamai sp. nov. may be read-

ily distinguished from its Sri Lankan congeners by a

combination of the following morphological and mer-

istic characteristics as well as color patterns: maximum

SVL 32.6 mm; dorsum with heterogeneous, smooth in-

termixed with weakly keeled granular scales; 2/2 supra-

nasals, one internasal, 2/2 postnasals; 3—4 enlarged post-

mentals; postmentals bounded by 5-6 chin scales; chin,

gular, pectoral, and abdominal scales smooth, subimbri-

cate; 21—22 belly scales across midbody; 6—7 well-devel-

December 2019 | Volume 13 | Number 2 | e216

Karunarathna et al.

Table 4. Meristic data of holotype and two paratypes of Cne-

maspis kotagamai sp. nov. from Bambaragala, Ratnapura Dis-

trict, Sri Lanka.

NMSL NMSL NMSL

2019.15.01 2019.15.02 2019.15.03

Counts Se a ee ee

Holotype Paratype Paratype

(Male) (Male) (Female)

FLSP (L/R) 6/7 6/6 7/7

SUP (L/R) 8/8 8/8 7/8

INF (L/R) FEL 8/7 7/7

INOS 31 29 31

PM 4 4 3

CHS 6 5 6

SUN (L/R) 2/2 2/2 2/2

PON (L/R) 2/2 2/2 2/2

INT l 1 l

SUS (L/R) 12/12 14/13 12/12

BET (L/R) 22122 21/19 21/22

CAS (L/R) 11/10 11/10 10/10

TLM (i) (L/R) 9/9 10/9 10/10

TLM (ii) (L/R) 12/12 12/11 12/12

TLM (iii) (L/R) 14/14 14/13 13/13

TLM (iv) (L/R) 15/15 14/14 14/13

TLM (v) (L/R) 12/12 11/12 12/12

PG 114 119 116

MBS 84 79 81

MVS 134 137 131

BLS 21 21 22

TLP (1) (L/R) 9/9 8/9 8/8

TLP (ii) (L/R) 14/14 13/14 14/14

TLP (iii) (L/R) 16/16 16/16 15/16

TLP (iv) (L/R) 17/17 17/18 18/17

TLP (v) (L/R) 16/16 15/15 16/15

PCP. 1 1 -

FP (L/R) 5/5 4/4 -

PFS (L/R) 12/13 11/12 -

DFS (L/R) 2/2 4/6 -

oped tubercles on posterior flank; 114—119 paravertebral

granules linearly arranged; one precloacal pore, 4-5 fem-

oral pores in males, separated by 11-13 unpored proxi-

mal femoral scales, 2—6 unpored distal femoral scales;

131-137 ventral scales; 79-84 midbody scales; subcau-

dals smooth, median row comprising an irregular series

of diamond-shaped, small scales; 7-8 supralabials; 7—8

infralabials; 13-15 total lamellae on 4" digit of manus,

and 17-18 total lamellae on 4" digit of pes.

Comparisons with other Sri Lankan species. Among

species of the C. kandiana clade sensu Agarwal et al.

(2017), Cnemaspis kotagamai sp. nov. differs by hav-

ing heterogeneous (versus homogeneous) dorsal scales

Amphib. Reptile Conserv.

from C. amith Manamendra-Arachchi et al. 2007, C.

gotaimbarai Karunarathna et al. 2019b, C. kumarasing-

hei Wickramasinghe and Munindradasa 2007, C. latha

Manamendra-Arachchi et al. 2007, and C. nandimithrai

Karunarathna et al. 2019b; it can also be diagnosed from

C. butewai Karunarathna et al. 2019b, C. kandiana (Ke-

laart, 1852), C. kivulegedarai Karunarathna et al. 201 9b,

C. menikay Manamendra-Arachchi et al. 2007, C. pava

Manamendra-Arachchi et al. 2007, C. pulchra Manamen-

dra-Arachchi et al. 2007, C. retigalensis Wickramasing-

he and Munindradasa 2007, C. samanalensis Wickrama-

singhe and Munindradasa 2007, C. si/vula Manamendra-

Arachchi et al. 2007, C. tropidogaster (Boulenger, 1885),

and C. upendrai Manamendra-Arachchi et al. 2007 by

having smooth (versus keeled) pectoral scales. The new

species differs from C. kallima Manamendra-Arachchi et

al. 2007 by having more midbody scales (79-84 versus

67-74), presence of more paravertebral granules (114—

119 versus 99-107), by having fewer precloacal pores

(1 versus 3-4), and having fewer tubercles on the poste-

rior flank (6—7 versus 12-15). It differs from C. ingero-

rum Batuwita et al. 2019 by having more ventral scales

(131-137 versus 88—95) and more paravertebral granules

(114-119 versus 93-101).

Among species of the C. podihuna clade sensu Agar-

wal et al. (2017), Cnemaspis kotagamai sp. nov. differs

by the absence of clearly enlarged, hexagonal or subhex-

agonal subcaudal scales from the following species: C.

alwisi Wickramasinghe and Munindradasa 2007, C. ans-

/emi Karunarathna and Ukuwela 2019, C. gemunu Bauer

et al. 2007, C. godagedarai de Silva et al. 2019, C. hiti-

hami Karunarathna et al. 2019b, C. kandambyi Batuwita

and Udugampala 2017, C. kohukumburai Karunarathna

et al. 2019b, C. molligodai Wickramasinghe and Munin-

dradasa 2007, C. nilgala Karunarathna et al. 2019, C.

phillipsi Manamendra-Arachchi et al. 2007, C. podihuna

Deraniyagala, 1944, C. punctata Manamendra-Arachchi

et al. 2007, C. rajakarunai Wickramasinghe et al. 2016,

C. rammalensis Vidanapathirana et al. 2014, and C. scal-

pensis (Ferguson 1877).

Description of Holotype (NMSL 2019.15.01). An adult

male, 29.8 mm SVL and 33.5 mm TAL. Body slender,

relatively long (TRL/SVL ratio 42.2%). Head relatively

small (HL/SVL ratio 28.0% and HL/TRL ratio 66.2%),

narrow (HW/SVL ratio 15.3% and HW/HL ratio 54.6%),

depressed (HD/SVL ratio 9.5% and HD/HL ratio 33.9%),

and distinct from neck. Snout relatively long (ES/HW ra-

tio 78.4% and ES/HL ratio 42.8%), less than twice eye

diameter (ED/ES ratio 52.5%), more than half length of

jaw (ES/JL ratio 72.1%), snout slightly concave in lateral

view; eye relatively small (ED/HL ratio 22.5%), larger

than the ear (EL/ED ratio 44.4%), pupil rounded; orbit

length greater than eye to ear distance (OD/EE ratio

127.8%) and length of IV digit of manus (OD/DLM IV

ratio 111.8%); supraocular ridges moderately developed;

ear opening small (EL/HL ratio 10.0%), deep, taller than

December 2019 | Volume 13 | Number 2 | e216

Three new species of Cnemaspis from Sri Lanka

wide, larger than nostrils; two rows of scales separate or-

bit from supralabials; interorbital distance is greater than

snout length (IO/ES ratio 101.7%), shorter than head

length (IO/HL ratio 43.5%); eye to nostril distance great-

er than the eye to ear distance (EN/EE ratio 109.5%).

Dorsal surface of the trunk with smooth scales inter-

mixed with weakly keeled heterogeneous granules, 114

paravertebral granules; 134 midventral scales, smooth; 84

midbody scales; 6/7 weakly developed tubercles on the

flanks; ventrolateral scales irregular, enlarged; granules

on snout smooth and raised, larger than those on interor-

bital and occipital regions; canthus rostralis nearly absent,

11/10 smooth oval scales from eye to nostril; scales of the

interorbital region circular and smooth; tubercles present

both on the sides of the neck and around the ear; ear open-

ing vertically oval, slanting from anterodorsal to postero-

ventral, 22/22 scales between anterior margin of the ear

opening and the posterior margin of the eye. Supralabials

8/8, infralabials 7/7, becoming smaller towards the gape.

Rostral scale wider than long, partially divided (80%) by

a median groove, contact with first supralabial. Nostrils

separated by 2/2 enlarged supranasals with one internasal;

no enlarged scales behind the supranasals. Nostrils oval,

dorsolaterally orientated, not in contact with first supra-

labials; 2/2 postnasals, smooth, larger than nostrils, par-

tially in contact with first supralabial.

Amphib. Reptile Conserv.

Wark

Fig. 5. General habitat of Cnemaspis kotagamai sp. nov. at Bambaragala isolated forest hill, Ratnapura District, Sri Lanka. (A) Rock

outcrop habitat, (B) abandoned cave building, and (C) deep and tall granite tunnel. Photos: Madhava Botejue.

Mental sub-rhomboid in shape, as wide as long, poste-

riorly in contact with four enlarged postmentals (smaller

than mental, and larger than chin scales); postmentals in

contact and bordered posteriorly by six unkeeled chin

scales (smaller than nostrils), in contact with the 1* in-

fralabial; ventral scales smaller than chin scales. Smooth,

rounded, juxtaposed scales on the chin and the gular re-

gion; pectoral and abdominal scales smooth, subimbri-

cate to imbricate towards precloacal region, abdominal

scales slightly larger than dorsals; 21 belly scales across

venter; smooth scales around vent and base of tail,

subimbricate; one precloacal pore; 5/5 femoral pores;

12/13 unpored proximal femoral scales on each side; 2/2

enlarged distal femoral scales. Regenerated tail of holo-

type a little longer than the snout-vent length (TAL/SVL

ratio 112.7%); hemipenal bulge greatly swollen (TBW

3.3 mm), heterogeneous scales on the dorsal aspect of

the tail directed backwards, spine-like tubercles present

at the base of tail; tail with 4—5 enlarged flattened obtuse

scales forming whorls; a large, blunt post-cloacal spur

on each side, dorsoventrally flattened and narrow; sub-

caudals smooth and small, subrhomboidal, arranged in a

single median series.

Forelimbs very short, slender (LAL/SVL ratio 11.4%

and UAL/SVL ratio 12.7%); hind limbs long, tibia shorter

than femur (TBL/SVL ratio 17.3% and FEL/SVL ratio

December 2019 | Volume 13 | Number 2 | e216

Karunarathna et al.

19.4%). Anterior surface of upper arm with keeled and

less imbricate scales; dorsal, posterior, and ventral sur-

face smooth, scales of the anterior surface twice as large

as those of the other surfaces; anterior and dorsal surfaces

of lower arm with keeled and less imbricate scales, ven-

tral and posterior surfaces with unkeeled imbricate scales,

scales on the anterior surface of upper arm and lower arm

twice the size of those of other aspects. Scales on dorsal

and ventral surfaces of femur smooth, those on anterior

and posterior surfaces keeled, scales on the ventral surface

twice the size of those of other aspects. Dorsal, anterior,

and posterior surfaces of tibia with keeled and weakly im-

bricate scales, ventral surface with smooth, subimbricate

scales, scales of the ventral surface twice as large as those

on other aspects. Dorsal and ventral surfaces of manus and

pes with keeled granules; dorsal surfaces of digits with

granular scales. Digits elongate and slender with inflected

distal phalanges, all bearing slightly recurved claws. Sub-

digital lamellae entire (except divided at first interphalan-

gial joint), unnotched; total lamellae on manus (left/right):

digit I (9/9), digit II (12/12), digit IM (14/14), digit IV

(15/15), digit V (12/12); total lamellae on pes (left/right):

digit I (9/9), digit II (14/14), digit II (16/16), digit IV

(17/17), digit V (16/16); interdigital webbing absent; rela-

tive length of left manual digits: I (1.4 mm), II (1.9 mm),

V (2.3 mm), II (2.5 mm), IV (2.9 mm); relative length of

left pedal digits: I (1.2 mm), I (2.1 mm), HI (2.9 mm), V

(3.5 mm), IV (3.8 mm).

Variation of the type series. The SVL of adult speci-

mens in the type series of Cnemaspis kotagamai sp. nov.

(n = 3) ranges from 29.8 to 32.6 mm; interorbital scales

29-31; supraciliaries above the eye 12—14; scales from

eye to tympanum 19-22; canthal scales 10—11; tubercles

on posterior flank 6—7; chin scales 5—6; ventral scales

131-137 (Tables 3-4); midbody scales 79-84; paraverte-

bral granules 114—119; belly scales across venter 21—22:

femoral pores in males 4—5:; unpored proximal femoral

scales in males 11-13; unpored distal femoral scales in

males 2—6; total lamellae on digit of the manus: digit I

(9-10), digit IT (11-12), digit HI (13-14), digit IV (13-

15), digit V (11-12); total lamellae on digit of the pes:

digit I (8-9), digit IT (13-14), digit HI (15-16), digit IV

(17-18), digit V (15-16).

Color of living specimens. Dorsum of head, body, and

limbs generally brown; one broad, yellow vertebral

stripe running form occiput to tail (Fig. 4); five irregular

blackish-brown paravertebral blotches present; occipital

area with a ‘W’-shaped dark marking. Tail dark brown

dorsally, with 11 faded black cross-bands; pupil circu-

lar and black with the surrounding margins yellow and

orange, supraciliaries yellowish; two black postorbital

stripes on each side; an oblique black line between the

eye and nostril on either side; supralabials and infralabi-

als yellowish with tiny black spots; chin and gular scales

dirty white, without dark spots; pectoral, abdominal, clo-

acal, and subcaudal scales immaculate cream; dorsum of

Amphib. Reptile Conserv.

limbs with faded black patches; manus and pes alternat-

ing black and cream-white crossbands.

Color of preserved specimens. Dorsally blackish-brown

with five distinct dark, irregular brown blotches (Fig. 3);

supralabials and infralabials dirty white; chin and gular

scales grey; ventral surface uniformly dirty white in col-

or, with some scales on thigh, tail base, and arms with

dark brown margins.

Etymology. The specific epithet is an eponym Latinized

(kotagamai) in the masculine genitive singular, honoring

prominent Sri Lankan scientist (ornithologist), Sarath Wi-

malabandara Kotagama (Emeritus Professor of the Uni-

versity of Colombo) for his valuable contributions towards

biodiversity conservation and management in Sri Lanka.

Distribution and naturalhistory. The type locality, Bam-

baragala forest (6.509086—6.522369°N and 80.742731—

80.759386°E; Ratnapura District, Sabaragamuwa Prov-

ince), is located in the lowland (southern intermediate

bioclimatic zone) where tropical moist semi-evergreen

forests comprise the dominant vegetation type (Guna-

tileke and Gunatileke 1990). The forest acreage is ~50 ha

and relatively isolated by anthropogenically-altered flat

lands. Bambaragala lies at an elevation of 110—178 m asl.

The mean annual rainfall of 1,500—2,000 mm Is received

mainly during the southwest monsoon (May—Septem-

ber), while the mean annual temperature 1s 27.8—29.6 °C.

Bambaragala is rich in granite rock caves with over 30

identified caves. Cnemaspis kotagamai sp. nov. appeared

to be a very rare species in Bambaragala, as only five

individuals were recorded during the survey. This species

was located in a granite cave on vertical surfaces, 4 m in

height, within the forested area (Fig. 5). The microhabitat

of C. kotagamai sp. nov. was poorly illuminated (light in-

tensity: 385-469 Lux), relatively moist (relative humid-

ity: 71-88%), canopy-shaded (canopy cover: 65—80%),

and relatively cool (ambient temperature: 29.8—31.3 °C

and substrate temperature: 27.8—28.6 °C). The new spe-

cles was sympatric with several other gecko species: Ge-

hyra mutilata, Hemidactylus depressus, H. frenatus, and

H. parvimaculatus. No eggs were observed.

Conservation status. Application of the IUCN Red List

criteria indicates that C. kotagamai sp. nov. is Critically

Endangered (CR), due to having an area of occupancy

(AOO) <10 km? (four locations, 0.13 km? in total assum-

ing a 100 m radius around the georeferenced location) and

an extent of occurrence (EOO) <100 km? (0.37 km/’) in

Sabaragamuwa Province [Applicable criteria B2-b (111)].

Remarks. Cnemaspis kotagamai sp. nov. most closely

resembles C. ingerorum (southern dry zone, ~85 m asl)

and C. kallima (northern wet zone, ~600 m asl) morpho-

logically, the type localities of these species are separated

by ~63 km (Sandagala in Tissamaharamaya) and ~115

km (Gammaduwa in Matale) straight line distances from

December 2019 | Volume 13 | Number 2 | e216

Three new species of Cnemaspis from Sri Lanka

Fig. 6. Holotype male of Cnemaspis dissanayakai sp. nov. (NMSL 2018.20.01). (A) Dorsal head, (B) lateral head, (C) ventral

head, (D) homogeneous dorsal scales, (E) scales on lateral surface of trunk, (F) smooth ventral scales, (G) cloacal characters with

precloacal pores and femoral pores, (H) lamellae on manus, (I) lamellae on pes, (J) smooth dorsal scalation of tail, (IX) lateral side

of tail, and (L) very small smooth subcaudals. Photos: Suranjan Karunarathna.

Bambaragala in Pallebedda (Fig. 1). Also see the com- —- Dissanayakage Diva-seri Hoona (Sinhala)

parison with other species for more details. Dissanayakavin Pahalpalli (Tamil)

Figs. 6-8; Tables 5—6

Cnemaspis dissanayakai sp. nov. Karunarathna, de

Silva, Madawala, Karunarathna, Wickramasinghe, urn:|sid:zoobank.org:act:7A BF9B28-7D04-4296-BDB5-ECEA07B1 F965

Ukuwela & Bauer

Dissanayaka’s Day Gecko (English) Holotype. NMSL 2019.20.01, adult male, 28.6 mm SVL

Amphib. Reptile Conserv. 334 December 2019 | Volume 13 | Number 2 | e216

Karunarathna et al.

Table 5. Morphometric data of holotype and two paratypes of

Cnemaspis dissanayakai sp. nov. from Dimbulagala, Polonna-

ruwa District, Sri Lanka.

NMSL NMSL NMSL

2019.20.01 2019.20.02 2019.20.03

Measurements Se ee

Holotype Paratype Paratype

(Male) (Male) (Female)

SVL 28.6 28.2 29.4

TRL 11.1 11.2 11.0

TW 55 5.4 oer

TD 3.6 3.4 33

TAL 31.1 31.2 34.4

TBW 25 2.7 2.8

TBD Di 23 Die

ED 1.9 19 1.9

OD 3.3 3.1 3.1

EN 2.4 2.4 2:3

ES 3.6 a7 3.6

SN 1.3 1.4 1.4

NW 0.2 O72 0.2

EE 2.5 oa 2,5

SA esa 12.9 12.8

EL 0.8 0.8 0.9

IO 3.6 3.6 mie

IE 3.8 3:7 se,

HL 9.0 8.9 8.9

HW 43 45 4.4

HD 2.4 25 2

JL 5D 5.6 5.6

IN 1.6 1.6 1.6

SED 8.1 8.2 8.2

UAL 4.6 4.6 4.6

LAL 4.2 4.2 4.2

PAL 3.2, Bee 33

DLM (1) 1.6 1.6 ies

DLM (ii) 1.8 1.8 1.9

DLM (iit) 2.8 2 Dal

DLM (iv) 33 Se 3.4

DLM (v) 25 2.4 2.6

FEL 5.6 5.6 36

TBL Saf Sl 5.5

HEL 3.9 3.8 3.8

DLP (1) 1.3 15 Ls

DLP (11) a2 3.2 343

DLP (iii) 3.6 3.6 SF

DLP (iv) 4 4l 4.2

DLP (v) are 3.8 3,7

Amphib. Reptile Conserv.

Table 6. Meristic data of holotype and two paratypes of Cne-

maspis dissanayakai sp. nov. from Dimbulagala, Polonnaruwa

District, Sri Lanka.

NMSL NMSL NMSL

2019.20.01 2019.20.02 2019.20.03

Counts

Holotype Paratype Paratype

(Male) (Male) (Female)

FLSP (L/R) 7/6 7/7 6/6

SUP (L/R) 7/7 7/7 7/7

INF (L/R) 7/7 7/7 7/7

INOS 31 29 29

PM 3 3 3

CHS ) 6 6

SUN (L/R) 212 2/2 2/2

PON (L/R) 1/1 1/1 1/1

INT 1 1 l

SUS (L/R) 16/16 16/17 16/15

BET (L/R) 22/23 21/21 21/22

CAS (L/R) 11/12 11/11 11/11

TLM (i) (L/R) 10/10 10/10 10/10

TLM (ii) (L/R) 13/12 1272 12/12

TLM (iit) (L/R) 13/13 12/13 12/12

TLM (iv) (L/R) a2i24 21/21 21/21

TLM (v) (L/R) 14/14 13/14 14/13

PG 105 107 105

MBS 98 94 95

MVS 118 120 119

BLS 17 LF 19

TLP (i) (L/R) 8/8 8/9 8/8

TLP (it) (L/R) 13/14 13/13 13/13

TLP (ii) (L/R) 16/16 16/15 16/16

TLP (iv) (L/R) 22/21 21-21 21/21

TLP (v) (L/R) 17/16 17/17 17/17

PCP 2 2 -

FP (L/R) 5/4 4/4 -

PFS (L/R) 10/10 11/10 -

DFS (L/R) 7/5 FT -

(Fig. 6), collected from a large granite cave in the shaded

forest of Dimbulagala, Polonnaruwa District, North-

Central Province, Sri Lanka (7.872931°N, 81.135569°E,

WGS1984; elevation 129 m; around 1600 hrs) on 12 July

2018 by Suranjan Karunarathna and Anslem de Silva.

Paratypes. NMSL 2019.20.02, adult female, 29.4 mm

SVL, and NMSL 2019.20.03, adult male, 28.2 mm SVL,

collected from moss covered granite cave in Dimbula-

gala, Polonnaruwa District, North-Central Province, Sri

Lanka (7.851358°N, 81.141675°E, WGS1984; elevation

135 m; around 1200 hrs) on 12 July 2018 by Suranjan

Karunarathna and Anslem de Silva.

December 2019 | Volume 13 | Number 2 | e216

Three new species of Cnemaspis from Sri Lanka

Diagnosis. Cnemaspis dissanayakai sp. nov., may be

readily distinguished from its Sri Lankan congeners by

a combination of the following morphological and mer-

istic characteristics: maximum SVL 29.4 mm; dorsum

with homogeneous, subconical granular scales; one in-

ternasal, 2/2 supranasals, 1/1 postnasals; 29-31 interor-

bital scales; 15—17 supraciliaries, 11-12 canthal scales,

21-23 eye to tympanum scales; three enlarged postmen-

tals; postmentals bounded by 6—7 chin scales; chin with

smooth granules, gular, pectoral, and abdominal scales

smooth, subimbricate; 17 belly scales across the venter;

6—7 well developed tubercles on posterior flank; 105—107

linearly arranged paravertebral granules; two precloacal

pores, 4—5 femoral pores on each side in males separated

by 10-11 unpored proximal femoral scales, 5-7 unpored

distal femoral scales; 118—120 ventral scales; 94—98 mid-

body scales; subcaudals smooth, median row small, in an

irregular series of diamond-shaped scales; 7/7 supralabi-

als; 7/7 infralabials; 21-22 total lamellae on 4" digit of

manus, and 21—22 total lamellae on 4" digit of pes.

Comparisons with other Sri Lankan species. Among

species of the C. kandiana clade sensu Agarwal et al.

(2017), Cnemaspis dissanayakai sp. nov. differs from C.

butewai, C. ingerorum, C. kallima, C. kandiana, C. ki-

vulegedarai, C. kotagamai sp. nov., C. menikay, C. pava,

C. pulchra, C. retigalensis, C. samanalensis, C. silvula,

C. tropidogaster, and C. upendrai by having homoge-

neous (versus heterogeneous) dorsal scales; from C.

amith by having smooth (versus keeled) pectoral scales;

from C. kumarasinghei, C. latha, and C. nandimithrai

by having more paravertebral granules (105-107 ver-

sus 61-68, 72-79, and 95-99, respectively), and from

by having more total lamellae on digit [V of manus and

digit IV of pes (21-22 versus 16-18, 17-18, and 19-20,

respectively); from C. gotaimbarai by having fewer

paravertebral granules (86-92 versus 117-121), fewer

ventral scales (107—114 versus 129-138), and fewer total

lamellae on digit IV of manus and digit IV of pes (15-16

versus 19-20).

Among species of the C. podihuna clade sensu Agar-

wal et al. (2017), Cnemaspis dissanayakai sp. nov. dif-

fers by the absence of clearly enlarged, hexagonal or

subhexagonal subcaudal scales from the following spe-

cies: C. alwisi, C. anslemi, C. gemunu, C. hitihami, C.

kandambyi, C. kohukumburai, C. molligodai, C. nilgala,

C. phillipsi, C. podihuna, C. punctata, C. rajakarunai, C.

rammalensis, and C. scalpensis.

Description of Holotype (NMSL 2019.20.01). An adult

male, 28.6 mm SVL, and 31.1 mm TAL. Body slender,

relatively short (TRL/SVL ratio 38.8%). Head relatively

long (HL/SVL ratio 31.5% and HL/TRL ratio 81.1%),

very narrow (HW/SVL ratio 15.1% and HW/HL ratio

48.0%), depressed (HD/SVL ratio 8.2% and HD/HL ra-

tio 26.2%), and distinct from neck. Snout relatively long

(ES/HW ratio 82.2% and ES/HL ratio 39.5%), less than

Amphib. Reptile Conserv.

twice eye diameter (ED/ES ratio 52.8%), more than half

length of jaw (ES/JL ratio 65.2%), snout slightly con-

cave in lateral view; eye relatively small (ED/HL ratio

20.8%), twice as large as the ear (EL/ED ratio 43.6%),

pupil rounded; orbit length greater than eye to ear dis-

tance (OD/EE ratio 131.0%) and also shorter than length

of IV digit of manus (OD/DLM IV ratio 99.7%); supra-

ocular ridges not prominent; ear opening very small (EL/

HL ratio 9.1%), deep, taller than wide, larger than nos-

trils; two rows of scales separate orbit from supralabials:

interorbital distance slightly shorter than snout length

(IO/ES ratio 99.7%), less than half of head length (1O/

HL ratio 39.4%); eye to nostril distance slightly shorter

than the eye to ear distance (EN/EE ratio 95.2%).

Dorsal surface of trunk with homogeneous, subconi-

cal granules; 105 paravertebral granules; 118 mid-ventral

scales, smooth; 98 midbody scales; 7/6 well developed

tubercles on flanks; ventrolateral scales not enlarged:

granules on snout strongly keeled, larger than those on

interorbital and occipital regions; canthus rostralis nearly

absent, 11/12 smooth round scales from eye to nostril;

scales of the interorbital region oval and smooth; 2/2

small and blunt tubercles present on sides of neck, and

around ear; ear opening vertically oval, backward slant-

ed, 22/23 scales between anterior margin of ear opening

and posterior margin of eye. Supralabials 7/7, infralabi-

als 7/7, becoming smaller towards the gape. Rostral scale

wider than long, partially divided (70%) by a median

groove, in contact with first supralabial. Nostrils sepa-

rated by 2/2 enlarged supranasals with one internasal; no

enlarged scales behind supranasals. Nostrils oval, dorso-

laterally orientated, not in contact with first supralabials;

1/1 postnasals, smooth, larger than nostrils, partially in

contact with first supralabial.

Mental subtriangular, as wide as long, posteriorly in

contact with three enlarged postmentals (smaller than

mental, and larger than chin scales); postmentals in con-

tact and bordered posteriorly by seven smooth chin scales

(smaller than nostrils), in contact only with 1* infralabials;

ventral scales smaller than chin scales; smooth, rounded,

juxtaposed scales on the chin and gular region; pectoral

and abdominal scales smooth, subimbricate to imbricate

towards precloacal region, abdominal scales slightly larger

than dorsals; 17 belly scales across venter; scales around

vent and base of tail smooth, subimbricate; two precloacal

pores; 5/4 femoral pores; 10/10 unpored proximal femo-

ral scales on each side; 7/5 enlarged distal femoral scales.

Original tail of holotype longer than snout-vent length

(TAL/SVL ratio 108.7%); tail base greatly swollen (TBW

2.5 mm), heterogeneous scales on dorsum of the tail di-

rected backwards, spine-like tubercles along tail; tail with

4-6 enlarged flattened obtuse scales forming whorls; a

small, blunt post-cloacal spur on each side, dorsoventrally

flattened and narrow; median series of smooth, irregular,

oval to rhomboid subcaudals.

Forelimbs moderately short, slender (LAL/SVL ra-

tio 14.7% and UAL/SVL ratio 15.9%); hind limbs long,

December 2019 | Volume 13 | Number 2 | e216

Karunarathna et al.

tibia barely longer than the femur (TBL/SVL ratio 19.7%

and FEL/SVL ratio 19.6%). Dorsal, anterior, and pos-

terior surfaces of upper arm and lower arm with keeled

and less imbricate scales than ventrals, ventral surfaces

smooth, less imbricate scales than ventrals, scales of the

anterior surface twice as large as those of the other sur-

faces. Scales on dorsal, posterior, and ventral surfaces of

femur smooth and granular, anterior surface with keeled

subimbricate scales, anterior surface twice as large as

those of the other aspects; dorsal, anterior and posterior

surfaces of tibia with keeled and subimbricate scales,

ventral scales smooth, subimbricate, twice as large as

those of the other limb surfaces. Manus and the pes with

keeled granules dorsally and ventrally; dorsum of dig-

its with granular scales; digits elongate and slender with

inflected distal phalanges, all bearing slightly recurved

claws. Subdigital lamellae entire (except divided at first

interphalangial joint), unnotched; total lamellae on ma-

nus (left/right): digit I (10/10), digit IT (13/12), digit I

(13/13), digit IV (22/21), digit V (14/14); total lamellae

on pes (left/right): digit I (8/8), digit II (13/14), digit II

(16/16), digit IV (22/21), digit V (17/16); interdigital

webbing absent; relative length of digits of left manus: I

(1.6 mm), II (1.8 mm), V (2.5 mm), III (2.8 mm), IV (3.3

‘ai &

mm); relative length of digits of left pes: I (1.5 mm), II

(3.2 mm), III (3.6 mm), V (3.9 mm), IV (4.1 mm).

Variation of the type series. The SVL of adult speci-

mens in the type series of Cnemaspis dissanayakai sp.

nov. (n = 3) ranges from 28.2 to 29.4 mm; interorbital

scales 29-31; supraciliaries above the eye 15—17; scales

from eye to tympanum 21—23; canthal scales 11-12; tu-

bercles on posterior flank 6—7; chin scales 6—7; ventral

scales 118-120; midbody scales 94-98; paravertebral

granules 105-107 (Tables 5-6); belly scales across ven-

ter 17-19; femoral pores 4-5; unpored proximal femor-

als 10-11; unpored distal femoral scales 5—7; total lamel-

lae on digit of the manus: digit I (8-9), digit II (13-14),

digit III (15-16), digit IV (21-22); total lamellae on digit

of the pes: digit I (8-9), digit IT (13-14), digit II (15-16),

digit IV (21-22), digit V (16-17).

Color of living specimens. Dorsum of the head, body,

and limbs generally dull brown, varying from light ma-

roon to light brown, five faded and irregular ‘W’-shaped

brown markings on the trunk; 4—5 cream vertebral blotch-

es (Fig. 7); an oblique black line between eye and nostrils

on either side, two straight, dark brown postorbital stripes

et |

Fig. 7. Holotype male of Cnemaspis dissanayakai sp. nov. (NMSL 2018.20.01) in life in-situ in Dimbulagala. (A) Dorsal view of

the full body, and (B) ventral view with dirty white coloration. Photos: Suranjan Karunarathna.

Amphib. Reptile Conserv.

337

December 2019 | Volume 13 | Number 2 | e216

Three new species of Cnemaspis from Sri Lanka

aes "i a E Pi. f

Fig. 8. General habitat of Cnemaspis dissan

ayakai sp. nov. at Dimbulagala isolated hill forest, Polonnaruwa District, Sri La

, . ai ae

nka.

(A) Complete view of the mountain, (B) abandoned ancient cave building in Kosgahaulpatha, and (C) deep and tall granite tunnel.

Photos: Madhava Botejue and Ashan Geeganage.

extend from eyes posteroventrally, and a faded spot pres-

ent in the occipital area. Tail grey-pink dorsally, with S—7

irregular faded brown cross-bands; pupil is circular and

black with the surrounding orange, with supraciliaries

being light brownish; supralabials dirty whitish dusted

with black; infralabials greyish dusted with black; mid-

gular scales are yellowish; pectoral, abdominal, cloacal,

and subcaudal scales white without markings; dorsum of

limbs with irregular brown patches and lines; manus and

pes with black and cream cross white stripes on dorsum.

Color of preserved specimens. Dorsum dark brown

with grey, faded indistinct irregular brown markings;

vertebral blotches cream. Venter dirty white with some

scales on throat, abdomen, thigh, tail base, and arms with

dark brown margins (Fig. 6).

Etymology. The specific epithet is an eponym Latinized

(dissanayakai) in the masculine genitive singular, honoring

Dissanayaka Mudiyanselage Karunarathna (born in Nilga-

la, Bibila) — father of the first author (Suranjan Karunara-

thna) for his encouragement, financial support for research,

and for allowing SK to pursue his interest in wildlife.

Distribution and natural history. The type local-

ity, Dimbulagala (7.843919-7.876344°N, 81.105603-—

Amphib. Reptile Conserv.

81.156442°E), situated in the Polonnaruwa District, North

Central Province (northeast dry bioclimatic zone) of Sri

Lanka, supports tropical dry-mixed evergreen forests (Gu-

natileke and Gunatileke 1990), and is ~1,000 ha in size.

The mean annual rainfall of 1,500—2,000 mm is received

mainly during the northeast monsoon (November—Febru-

ary). The mean annual temperature of the area is 28.9—30.2

°C, and its elevation range is 120-250 m asl. According

to preliminary investigations, Cnemaspis dissanayakai sp.

nov. appeared to be very rare in Dimbulagala. The survey

of 35 ha recorded two (+ 0.1) geckos per surveyor-hour of

effort. This species was restricted to rocky surfaces and

granite caves in shaded forested areas, and old abandoned

buildings inside the forest (Fig. 8). These microhabitats

were well-shaded (light intensity: 594-648 Lux), rela-

tively humid (relative humidity: 65-90%), and moderately

warm (ambient temperature: 30.2—31.9 °C and substrate

temperature 27.5—28.6 °C). The new species was observed

to occur in sympatry with the following gecko species:

Calodactylodes illingworthorum, Gehyra mutilata, Hemi-

dactylus depressus, H. frenatus, H. hunae, H. parvimac-

ulatus, and H. triedrus. Older and newly laid eggs were

observed in granite rock crevices, usually laid in clusters

of three. The eggs were pure white in color and almost

spherical in shape (mean diameter 4.9 + 0.02 mm), with a

slightly flattened side attached to the rocky substrate.

December 2019 | Volume 13 | Number 2 | e216

Karunarathna et al.

_ - ‘3 A

re r

Fig. 9. Holotype male of Cnem

aspis kawminiae sp. nov. (NMSL 2018.18.01). (A) Dorsal head, (B) lateral head, (C) ventral head,

Haat

(D) homogeneous dorsal scales, (E) scales on lateral surface of trunk, (F) smooth ventral scales, (G) cloacal characters with precloa-

cal pores and femoral pores, (H) lamellae on manus, (I) lamellae on pes, (J) smooth dorsal scalation of tail, (IX) lateral side of tail,

and (L) very small subcaudals. Photos: Suranjan Karunarathna.

Conservation status. Application of the IUCN Red

List criteria indicates that C. dissanayakai sp. nov. is

Critically Endangered (CR) due to having an area of

occupancy (AOO) <10 km? (four locations, 0.13 km? in

total assuming a 100 m radius around the georeferenced

location) and an extent of occurrence (EOO) <100 km?

(4.08 km?) in North Central Province [Applicable crite-

ria B2-b (ii1)].

Amphib. Reptile Conserv.

Remarks. Cnemaspis dissanayakai sp. nov. most closely

resembles C. kumarasinghei (east intermediate zone) and

C. latha (southern intermediate zone) morphologically.

The type localities of these species are separated by ~105

km (Maragala in Monaragala, ~500 m asl) and ~90 km

(Bandarawela in Badulla, ~700 m asl) straight line dis-

tances from Dimbulagala in Polonnaruwa (Fig. 1). Also

see the comparison with other species for more details.

December 2019 | Volume 13 | Number 2 | e216

Table 7. Morphometric data of holotype and two paratypes of

Cnemaspis kawminiae sp. nov. from Mandaramnuwara, Nu-

wara-Eliya District, Sri Lanka.

Measurements

SVL

TRL

TAL

TBW

TBD

SED

UAL

LAL

PAL

DLM (i)

DLM (ii)

DLM (iii)

DLM (iv)

DLM (v)

FEL

TBL

HEL

DLP (i)

DLP (ii)

DLP (iii)

DLP (iv)

DLP (v)

Amphib. Reptile Conserv.

Three new species of Cnemaspis from Sri Lanka

NMSL NMSL NMSL

2019.18.01 2019.18.02 2019.18.03

Holotype Paratype Paratype

(Male) (Male) (Female)

33.7 33.2 5556,

16.4 14.9 16.2

RO) 5.4 5.4

3.4 3.2 3:3

36.1 42.7 38.0

3.4 3.4 3:2

2:9 2.8 2.7

1.5 1.5 1.4

3.1 29 Dd

3.1 SZ. Sl

4.3 45 4.8

2 1.3 1s

0.3 0.3 0.3

2 Sie 3,2

14.9 14.8 14.9

0.9 0.9 0.9

2.9 29) 2)

37 af 3:3

9.9 9.4 10.4

3:5 5.3 a2

3.8 3.6 3.6

5.9 5.8 5.8

1.3 1.3 1.3

8.3 8.2 8.0

45 43 43

45 4.5 4.5

4.6 4.2 4.2

2.1 2.2 2.2

235 2S, 2.6

2.6 27 2.6

3.0 29) 3.0

Ze zie 2.2

6.5 6.3 6.5

6.1 6.1 6.1

5:3 5.8 6.1

2.1 221 2.1

3.0 2 3.0

3.5 3.5 3.5

3.8 3.8 3.8

3.6 3.8 3.6

Table 8. Meristic data of holotype and two paratypes of Cne-

maspis kawminiae sp. nov. from Mandaramnuwara, Nuwara-

Eliya District, Sri Lanka.

NMSL NMSL NMSL

2019.18.01 2019.18.02 2019.18.03

Counts

Holotype Paratype Paratype

(Male) (Male) (Female)

FLSP (L/R) 8/7 77 8/7

SUP (L/R) 8/8 8/8 8/7

INF (L/R) 7/7 8/7 77

INOS eA | 20 22

PM 3 3 3

CHS 5 a 5

SUN (L/R) 2/2 2/2 2/2

PON (L/R) 2/2 2/2 2/2

INT 1 1 |

SUS (L/R) 10/10 10/10 10/9

BET (L/R) 22/20 21/21 22/20

CAS (L/R) 10/11 10/10 10/10

TLM (i) (L/R) 10/9 10/9 10/10

TLM (ii) (L/R) 13/13 12/12 13/12

TLM (iii) (L/R) 14/13 14/14 14/14

TLM (iv) (L/R) 15/14 15/15 14/14

TLM (v) (L/R) 13/14 14/14 14/13

PG 89 92 86

MBS 78 76 76

MVS 107 108 114

BLS 21 17 19

TLP (i) (L/R) 9/9 10/9 10/10

TLP (ii) (L/R) 12/13 12/12 12/12

TLP (iii) (L/R) 16/15 16/16 15/15

TLP (iv) (L/R) 16/16 15/16 15/16

TLP (v) (L/R) 14/14 14/15 15/15

PCP 2 2 -

FP (L/R) 4/4 4/4 2

PFS (L/R) 12/11 12/13 2

ROME __ Gis _ Ee A _t

Cnemaspis kawminiae sp. nov. Karunarathna, de Silva,

Gabadage, Karunarathna, Wickramasinghe, Ukuwela &

Bauer

Kawmini’s Day Gecko (English)

Kawminige Divaseri Hoona (Sinhala)

Kawminivin Pahalpalli (Tamil)

Figs. 9-11; Tables 7-8

urn:lsid:zoobank.org:act: 12E14150-D66F-471C-98B1-E805C5A 9244F

Holotype. NMSL 2019.18.01, adult male, 33.7 mm SVL

(Fig. 9), collected from a moss-covered granite wall in

Mandaramnuwara, bordering Pidurutalagala Mountain

range, Nuwara-Elitya District, Central Province, Sri

340 December 2019 | Volume 13 | Number 2 | e216

Karunarathna et al.

Lanka (7.033558°N, 80.798794°E, WGS1984; elevation

1,600 m asl, around 1100 hrs) on 14 December 2018 by

Suranjan Karunarathna and Anslem de Silva.

Paratypes. NMSL 2019.18.02, adult male, 33.2 mm SVL

and NMSL 2019.18.03, adult female, 35.2 mm SVL, col-

lected from a small granite cave Mandaramnuwara, bor-

dering Pidurutalagala Mountain, Nuwara-Eliya District,

Central Province, Sri Lanka (7.020600°N, 80.788639°E,

WGS1984; elevation 1,658 m asl, around 1400 hrs) col-

lected on 14 December 2018 by Suranjan Karunarathna

and Anslem de Silva.

Diagnosis. Cnemaspis kawminiae sp. nov., may be read-

ily distinguished from its Sri Lankan congeners by a

combination of the following morphological and meristic

characteristics: maximum SVL 35.2 mm; dorsum with

homogeneous flat granular scales; one internasal, 2/2 su-

pranasals and 2/2 postnasals; 20—22 interorbital scales;

9-10 supraciliaries, 10—11 canthal scales, 20-22 eye to

tympanum scales; three enlarged postmentals; postmen-

tals bounded by five chin scales; chin with smooth and

round granules, gular, pectoral, and abdominal scales

smooth, subimbricate; 17—21 belly scales across the ven-

ter; 7-8 weakly developed tubercles on posterior flank:

86-92 linearly arranged paravertebral granules; two pre-

cloacal pores in males, 4/4 femoral pores on each side

in males separated by 11—13 unpored proximal femoral

scales, 6-7 unpored distal femoral scales; 107-114 ven-

tral scales; 76-78 midbody scales; subcaudals smooth,

median row small, in an irregular series of sub-rhomboid

shaped scales; 7-8 supralabials; 7-8 infralabials; 14—15

total lamellae on 4" digit of manus, and 15-16 total la-

mellae on 4" digit of pes.

Comparisons with other Sri Lankan species. Among

species of the C. kandiana clade sensu Agarwal et al.

(2017), Cnemaspis kawminiae sp. nov. differs from C.

butewai, C. ingerorum, C. kallima, C. kandiana, C. ki-

vulegedarai, C. kotagamai sp. nov., C. menikay, C. pava,

C. pulchra, C. retigalensis, C. samanalensis, C. silvula, C.

tropidogaster, and C. upendrai by having homogeneous

(versus heterogeneous) dorsal scales; from C. amith by

having smooth (versus keeled) pectoral scales; from C.

gotaimbarai, C. kumarasinghei, and C. dissanayakai

sp. nov. by having fewer ventral scales (107-114 versus

129-138, 120-134 and 118-120, respectively), and also

from C. kumarasinghei and C. dissanayakai sp. nov. by

having fewer midbody scales (76—78 versus 87-94 and

94-98, respectively), from C. gotaimbarai by having

fewer paravertebral granules (86—92 versus 117-121),

from C. latha by having more paravertebral granules

(86—92 versus 72—79), and more belly scales (17-21 ver-

sus 13-15); from C. nandimithrai by having fewer belly

scales (17-21 versus 25-27) and by having fewer total

lamellae on digit IV of pes (15—16 versus 19-20).

Amphib. Reptile Conserv.

The new species, Cnemaspis kawminiae sp. nov., also

clearly differs from the following species of the C. po-

dihuna clade sensu Agarwal et al. (2017): C. alwisi, C.

anslemi, C. gemunu, C. godagedarai, C. hitihami, C.

kandambyi, C. kohukumburai, C. molligodai, C. nilgala,

C. phillipsi, C. podihuna, C. punctata, C. rajakarunai, C.

rammalensis, and C. scalpensis by the absence (versus

presence) of clearly enlarged, hexagonal or subhexago-

nal subcaudal scales.

Description of Holotype (NMSL 2019.18.01). An adult

male, 33.7 mm SVL, and 36.1 mm TAL. Body slender,

relatively long (TRL/SVL ratio 48.7%). Head relatively

small (HL/SVL ratio 29.4% and HL/TRL ratio 60.4%),

relatively broad (HW/SVL ratio 16.4% and HW/HL

ratio 55.6%), weakly depressed (HD/SVL ratio 11.2%

and HD/HL ratio 38.1%), and distinct from neck. Snout

relatively short (ES/HW ratio 78.4% and ES/HL ratio

43.6%), slightly less than three times eye diameter (ED/

ES ratio 34.4%), more than half length of jaw (ES/JL ra-

tio 73.3%), snout slightly concave in lateral view; eye

very small (ED/HL ratio 15.0%), larger than ear (EL/ED

ratio 59.7%), pupil rounded; orbit length slightly smaller

than eye to ear distance (OD/EE ratio 97.2%) and lon-

ger than length of IV digit of manus (OD/DLM IV ra-

tio 105.4%); supraocular ridges weakly prominent; ear

opening very small (EL/HL ratio 9.0%), deep, taller than

wide, larger than nostrils; two rows of scales separate or-

bit from supralabials; interorbital distance less than snout

length (IO/ES ratio 67.7%), head length three times lon-

ger than interorbital distance ([O/HL ratio 29.5%); eye to

nostril distance subequal to eye to ear distance (EN/EE

ratio 98.1%).

Dorsal surface of trunk with homogeneous, flat granu-

lar and smooth scales; 112 paravertebral granules; 149

midventral scales, keeled; 69 midbody scales; 6/6 well

developed tubercles on flanks; ventrolateral scales ir-

regularly enlarged; granules on snout strongly keeled,

larger than those on interorbital and occipital regions;

canthus rostralis nearly absent, 9/10 smooth round scales

from eye to nostril; scales of the interorbital region oval

and smooth; small and blunt tubercles present on sides of

neck, and around ear; ear opening vertically oval, slant-

ing from anterodorsal to posteroventral, 20/19 scales be-

tween anterior margin of ear opening and the posterior

margin of eye. Supralabials 8/7, infralabials 7/7, becom-

ing smaller towards the gape. Rostral scale wider than

long, partially divided (90%) by a median groove, in con-

tact with first supralabial. Nostrils separated by 2/2 en-

larged supranasals with one internasal, 2/2 postnasals; no

enlarged scales behind supranasals. Nostrils oval, dorso-

laterally orientated, not in contact with first supralabials.

Mental subtriangular, as wide as long, posteriorly in

contact with three enlarged postmentals (smaller than

mental, and lager than chin scales); postmentals in con-

tact and bordered posteriorly by five smooth chin scales

December 2019 | Volume 13 | Number 2 | e216

Three new species of Cnemaspis from Sri Lanka

(smaller than nostrils), in contact only with 1 and 2"4

infralabials; ventral scales smaller than chin scales.

Smooth, oval, juxtaposed scales on the chin and gular

region; pectoral and abdominal scales smooth, subim-

bricate to imbricate towards precloacal region, abdomi-

nal scales slightly larger than dorsals; 21 belly scales

across venter; scales around vent and base of tail smooth,

subimbricate; two precloacal pores; 4/4 femoral pores;

12/11 unpored proximal femoral scales on each side; 7/6

enlarged distal femoral scales. Original tail of holotype

longer than snout-vent length (TAL/SVL ratio 106.9%);

hemipenial bulge greatly swollen (TBW 3.4 mm), homo-

geneous scales on dorsum of the tail directed backwards,

spine-like tubercles along tail; tail with 4—5 enlarged flat-

tened obtuse scales forming whorls; a large, blunt post-

cloacal spur on each side, dorsoventrally flattened and

narrow; a single median series of smooth, irregular, oval

to rhomboid subcaudals.

Forelimbs very short, slender (LAL/SVL ratio 13.3%

and UAL/SVL ratio 13.3%), upper arm and lower arm

equal in size; hind limbs long, tibia slightly shorter

than femur (TBL/SVL ratio 18.0% and FEL/SVL ratio

19.3%). Dorsal, anterior, and posterior surfaces of upper

arm and lower arm with keeled and less imbricate scales

than ventral scales, ventral surfaces with smooth scales,

scales of the anterior surface twice as large as those of the

other aspects. Scales on anterior and posterior surfaces of

femur keeled, dorsal and ventral scales smooth, ventral

scales twice as large as those of the other limb surfaces.

Scales on dorsal, anterior, and posterior surfaces of tibia

keeled, ventral scales smooth, anterior scales twice as

large as those of the other limb surfaces. Manus and pes

with smooth granules dorsally and ventrally; dorsum of

digits with conical granular smooth scales. Digits elon-

gate and slender with inflected distal phalanges, all bear-

ing slightly recurved claws. Subdigital lamellae entire

(except divided at first interphalangial joint), unnotched;

total lamellae on manus (left/right): digit I (10/9), dig-

it It (13/13), digit III (14/13), digit TV (15/14), digit V

(13/14); total lamellae on pes (left/right): digit I (9/9),

digit II (12/13), digit HI (16/15), digit ITV (16/16), digit

V (14/14); interdigital webbing absent; relative length of

left manual digits: I (2.1 mm), V (2.2 mm), II (2.5 mm),

IIT (2.6 mm), IV (3.0 mm); relative length of left pedal

digits: 1 (2.1 mm), II (3.0 mm), III (3.5 mm), V (3.6 mm),

IV (3.8 mm).

Variation of the type series. The SVL of adult speci-

mens in the type series of Cnemaspis kawminiae sp. nov.

(n = 3) ranges from 33.2 to 35.6 mm; interorbital scales

20-22; scales from eye to tympanum 20-22; canthal

scales 10-11; supraciliaries 9-10; tubercles on posterior

flank 7—8; ventral scales 107—114 (Tables 7-8); midbody

scales 76-78, paravertebral granules 86—92, belly scales

across venter 17—21; unpored proximal femorals 11-13

in males, unpored distal femoral scales 6—7 in males; to-

tal lamellae on digit of the manus: digit I (9-10), digit

Amphib. Reptile Conserv.

II (12-13), digit HI (13-14), digit TV (14-15), digit V

(13-14); total lamellae on digit of the pes: digit I (9-10),

digit I (12-13), digit I (15-16), digit IV (15-16), digit

V (14-15).

Color of living specimens. Dorsal body, limb, and tail

generally light grey to brown, with an oblique black line

in the interorbital area, also between eye and nostril; a

wide ‘W’-shaped, black patch on the occipital area with

two median cream-white spots; four scattered, double

“W’-shaped brownish markings on the dorsum of the

trunk with tiny irregular stripes, and ten grey brownish

blotches along the tail (Fig. 10). Lateral side of limbs and

body grey-brown with scattered black spots, and cream

colored lateral conical tubercles on tail and trunk. Three

straight, dark brown postorbital stripes-downwards and

upwards; supraciliaries and nasals greyish brown. Pupil

circular and black with the surrounding scales yellowish

brown, supralabials and infralabials with a median cream

spot. Ventral surfaces of head, body, and limbs beige to

cream, but gular area covered in tiny black spots; ventral

surface of tail cream colored.

Color of preserved specimens. Dorsum cinnamon

brown, with faded double ‘W’-shaped patches on dor-

sum, irregular tiny brown dots on head; faded brown line

between eye and nostrils on both sides, and three brown

postorbital stripes on either side (Fig. 9); venter dirty

white with some scales on throat, abdomen, thigh, tail

base, and arms with dark brown dots.

Etymology. The specific epithet is an eponym Latinized

(kawminiae) in the feminine genitive singular, honoring

Hadunneththi Kawmini Mendis — mother of the first au-

thor (Suranjan Karunarathna) for her unconditional love,

generous support, and financial support for research.

Distribution and natural history. The type local-

ity, Mandaramnuwara (7.020103—7.039953°N and

80.768794—80.807014°E) in the east wet bioclimatic

zone is located at the northern part of Pidurutalagala

mountain range (Fig. 11). This area supports tropical

montane forest vegetation (Gunatilleke and Gunatilleke

1990) with wet evergreen forest. The core study area was

~300 ha in size, at an elevation range ~1,400—1,800 m

asl and annual temperature of 27.4—28.9 °C. Cnemaspis

kawminiae sp. nov. was not abundant in the study area

as only five (+0.1) geckos per surveyor-hour were found

in Mandaramnuwara, This species was found on moss

covered boulders and rock surfaces in forested areas and

well-shaded home gardens with ample woody tree cover

(light intensity: 486-592 Lux); as well as rock walls and

rock crevices along roads. These habitats were very wet

and cool (ambient temperature: 24.2—26.5 °C, substrate

temperature: 26.7—28.3 °C, canopy cover: 70-85% and

relative humidity: 74-92%). The mean annual rainfall of

3,000—4,000 mm is received mainly during the southwest

December 2019 | Volume 13 | Number 2 | e216

Karunarathna et al.

Fig. 10. Holotype male of Cnemaspis poe Sp. nov. . (NMSL 2018, 18.01) in life in-situ. (I Dorsal view oi the full body, and

(B) dorsolateral view with labial coloration. Photos: Madhava Botejue.

monsoon (May—September). A total of 26 females, 11

males, and eight juveniles of this species were observed

from twelve sites in the Mandaramnuwara area. Dur-

ing July to September, hatchlings, juveniles, and gravid

females carrying one or two eggs were observed. Eggs

were pure white (mean diameter 5.2 + 0.02 mm), and al-

most completely round in shape with a slightly flattened

side which was often the side attached to the substrate or

between the eggs.

Conservation status. Application of the IUCN Red

List criteria indicates that C. kawminiae sp. nov. 1s

Critically Endangered (CR) due to having an area of

occupancy (AOO) <10 km? (four locations, 0.13 km? in

total assuming a 100 m radius around the georeferenced

location) and an extent of occurrence (EOO) <100 km?

(2.32 km’) in Central Province [Applicable criteria

B2-b (ii1)].

Amphib. Reptile Conserv.

Remarks. Cnemaspis kawminiae sp. nov. most closely

resembles C. kumarasinghei (east intermediate zone)

and C. gotaimbarai (northeast dry zone) morphological-

ly. The type localities of these species are separated by

~80 km (Maragala in Monaragala, ~500 m asl) and ~44

km (Kokagala in Padiyathalawa, ~300 m asl) straight

line distances from Mandaramnuwara (~1,500 m asl) in

Nuwara-Eliya District (Fig. 1). Also see the comparison

with other species for more details.

Discussion

The recent renaissance in the taxonomy and systematics

of genus Cnemaspis has led to a notable increase in spe-

cies richness, particularly from south and south-eastern

Asia, including the Indo-Malayan mainland as well as

Indian-oceanic and south-pacific islands (Iskandar et al.

2017; Riyanto et al. 2017; Wood et al. 2017; Karunara-

December 2019 | Volume 13 | Number 2 | e216

Three new species of Cnemaspis from Sri Lanka

Fig. 11. General habitats of Cnemaspis

view of the granite hill at roadside, (B) small granite cave close to the stream, and (C) granite rock wall along the road. Photos:

Madhava Botejue.

thna et al. 2019b; Uetz et al. 2019a). With over 160 spe-

cies, Cnemaspis is considered the second-most speciose

gecko genus in the world, after Cyrtodactylus (Grismer

et al. 2014; Present paper). With the inclusion of the three

new species described here, species richness of Cnemas-

pis, the most species-rich reptile genus of Sri Lanka, rises

to 36 (13 species described in year 2019). Similar phy-

logenetic radiations with high degrees of endemism via

complex evolutionary processes have been documented

for snakes, other Gekkonid squamates, and amphib-

ians of Sri Lanka (Bauer et al. 2010; Pyron et al. 2013b;

Meegaskumbura et al. 2019). Our study further bolsters

the notion that Sri Lanka is a hotspot for reptile diver-

sity and endemism (Bossuyt et al. 2004). Since all three

currently described members of this genus are endemic

to the island, Cnemaspis exhibits the greatest degree of

genus-level endemism in Sri Lanka. Sri Lankan Cnemas-

pis species represent two distinct evolutionary lineages,

the kandiana and podihuna clades (Agarwal et al. 2017;

Karunarathna et al. 2019b).

The three new species described in this paper have not

been included in any previous phylogenies of the genus

Amphib. Reptile Conserv.

(Bauer et al. 2007; Agarwal et al. 2017; Karunarathna

et al. 2019b). All of these new species (C. dissanayakai

sp. nov., C. kawminiae sp. nov., and C. kotagamai sp.

nov.) were assigned to the C. kandiana clade based on

the presence of small and irregularly shaped subcaudal

scales (see Karunarathna and Ukuwela 2019). Howev-

er, more in-depth phylogenetic studies are necessary to

confirm the placement of these three new species within

this clade and subgroups (Table 9). Hence, we strongly

recommend broader and more robust molecular phyloge-

netic studies on Cnemaspis species, as well as on other

gecko species, to identify the true richness within the is-

land. Almost all Sri Lankan Cnemaspis species are found

within relatively cool, moist habitats (ambient tempera-

ture: 24.2—32.3 °C; substrate temperature: 25.2—28.7 °C;

relative humidity: 68-92%), with relatively high levels

of canopy cover and high-profile mature trees, and shady

(canopy cover: 60-90%; light intensity: 385-821 Lux)

environments with tall large trees (Karunarathna et al.

2019b). Moreover, all of these new species were found

in granite caves or in association with rocky substrates.

Such aspects of natural history and microhabitat selec-

December 2019 | Volume 13 | Number 2 | e216

Karunarathna et al.

10Ysul

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December 2019 | Volume 13 | Number 2 | e216

345

Amphib. Reptile Conserv.

Three new species of Cnemaspis from Sri Lanka

tion of these new species are com-

parable to other congeners, which

imply niche conservatism among

divergent lineages.

The bulk of these rupicolous

geckos are restricted to cool,

moist, shady granite caves and

rock walls or under granite boul-

ders. According to our findings

these geckos prefer narrow (~3—4

mm), long (~100-400 mm), and

deep (~20—180 mm) crevices as

refugia and oviposition sites. In

several surveyed locations with

granite caves, we were unable to

find Cnemaspis species due to the

lack of tall shady trees and ad-

equately cool temperatures (sub-

strate temperature: 25.2—28.7°C).

A majority (66.7%) of Cnemaspis

Species are restricted to the wet

bioclimatic zones of Sri Lanka

and are point-endemic microhabi-

tat specialists where distribution

ranges are limited to <100 km”.

The restricted distribution could

be an artifact of the limited avail-

ability of caves and similar mi-

croenvironments with granite or

rock-based substrates. The high

species richness in Sri Lanka may

be accounted by the possibility

of multiple, independent coloni-

zation events from the Indian

mainland with subsequent, geo-

graphically-isolated in-situ spe-

ciation. The majority of the Indian

Cnemaspis species have not been

comparatively analyzed alongside

the Sri Lankan species (Agarwal

et al. 2017; Karunarathna et al.

2019b). The three new species

described here are recorded from

isolated locations in wet, interme-

diate, and dry bioclimatic zones of

Sri Lanka. Of these, C. kawminiae

sp. nov. is described from the wet

zone montane region; C. kotaga-

mai sp. nov. 1s described from the

intermediate zone lowland; and C.

dissanayakai sp. nov. is described

from the dry zone lowland (Table

10). The record of C. kawminiae

sp. nov. from Mandaramnuwara

is noteworthy, as this is a high-

altitude location nestled in the

central highlands (1,400—1,800 m

—_~

—}

Group (2) — podihuna

ss oo | se [se [se [one] ve [oe forlorn] + Poe [ef mm f 9

soo [ [se [oy [ons] oe [re [oem] = [ore foes] = Pos [oo [om | os

se for] se [se [oy [one] sn] v9 [war [ sa] ve fem [al es | — [orem | 2m

C. nandim-

C. anslemi

C. hitihami

C. kohukum-

C. nilgala

C. phillipsi

C. punctata

C. rajakarunai

C. rammalen-

C. molligodai

C. podihuna

SUP — Supralabials, INF — Infralabials, PG — Paravertebral granules, IFS — Interfemoral scales, FLSP — Flank spines, PCP — Precloacal pores, FP — Femoral pores, HET — Heterogeneous, HOM

C. gemunu

— Homogeneous, KD — Keeled, SM — Smooth.

Table 9 (continued). Key characters of 36 currently known Cnemaspis species in Sri Lanka. Abbreviations: mm — Millimeters, SVL — Maximum Snout to vent length, SUB — Subcaudals,

Amphib. Reptile Conserv. 346 December 2019 | Volume 13 | Number 2 | e216

Karunarathna et al.

i ‘ £ i

Fe. ' F

1 Poe ah be T

5 1 Bot etal Nag ei aes

; ‘ ra = a - 3 = f

L -. . faire A fae f :

s ‘ ; . as * Sept |

a Ls, re, Ea - : st af

ies it ah. 4} 4 ae.

AE SSS ke

Fig. 12. Threats to the isolated hill forests and Cnemaspis species in study areas in Sri Lanka. (A) Illegal forest clearing, (B) fire-

wood collection for tea factory, (C) granite mining activities, (D) agricultural fields in slopy areas, (E) tea plantation and highly

crowded anthropogenic habitat, and (F) a landslide in mountain areas. Photos: Suranjan Karunarathna and Madhava Botejue.

asl), making this the 4" species in the genus found at an

elevation above 1,000 m asl.

Bambaragala and Dimbulagala are isolated residual

mountains and rock outcrops embedded within a forest

and features granite caves incorporated with historical

Buddhist monasteries, whereas Mandaramnuwara is a

mixture of forested areas and rural human habitation. All

these habitats are susceptible to human-induced habitat

degradation, including clear cutting and timber felling,

forest fragmentation, granite mining, tea cultivation (Fig.

12), rubber cultivation, vegetable farming, invasive spe-

cies, human settlements, road and other infrastructure

development, and waste disposal (see Karunarathna et

Amphib. Reptile Conserv.

al. 2017). Bambaragala, situated in Ratnapura District of

Sabaragamuwa Province, is the most vulnerable habitat

as it is a small forested rock outcrop (~50 ha) located

amidst a rapidly urbanizing landscape; where a part of

the rock outcrop is currently undergoing mining, mak-

ing C. kotagamai sp. nov. the most endangered amongst

these new species. However, many such habitats are

somewhat protected due to the presence of Buddhist

monasteries which serve as refugia for reptiles and oth-

er faunal groups, and it is imperative to conserve these

habitats to protect the island’s unique biodiversity (Ama-

rasinghe et al. 2016; Edirisinghe et al. 2018; Karunara-

thna et al. 2019a). Sri Lanka’s tropical humid wet zone is

December 2019 | Volume 13 | Number 2 | e216

Three new species of Cnemaspis from Sri Lanka

globally recognized for its exceptionally high biodiver-

sity and endemism (Bossuyt et al. 2004). Nonetheless,

the new species reported here and in previous studies on

the same genus continue to illustrate the undocumented

diversity of Cnemaspis that also occurs within the dry

and intermediate bioclimatic zones (Batuwita et al. 2019;

Karunarathna et al. 2019b).

Most of the Cnemaspis species from the dry and in-

termediate climatic zones of Sri Lanka are, however, re-

stricted to small isolated habitats scattered over the low-

lands (Batuwita et al. 2019; Karunarathna et al. 2019a,b).

The presence of granitic caves and the humid forest cover

surrounding the caves seem to serve as ideal refugia for

these geckos with narrow, specialized ecological niches.

It is very likely that future studies on the biogeography of

Cnemaspis in Sri Lanka will highlight the importance of

these isolated habitats in generating and maintaining the

diversity of these unique groups of geckos in the island

(Karunarathna and Amarasinghe 2011; Amarasinghe et

al. 2016). At the same time, it is important to note that the

point endemic species described here, which are highly

sensitive to changes in the habitat, would be severely af-

fected by habitat degradation. Hence, past and present

studies have emphasized the importance of conserving

such isolated habitats throughout the country (Karunara-

thna and Amarasinghe 2013; Gabadage et al. 2018).

Though traditional conservation strategies usually target

extensive natural habitats to maximize biodiversity con-

servation, our studies indicate that these small isolated

habitats also deserve the immediate attention of conser-

vation authorities. Thus, we believe our findings on these

geckos and their granite cave and rock-associated habi-

tats add a new dimension to the biodiversity conservation

of Sri Lanka.

Relative

humidity

Light

intensity

(Lux)

469

385

448

455

648

594

617

625

592

566

486

578

Substrate

temperature

29.8 °C 28.1 °C

Ambient

temperature

30.6 °C 28.6 °C

29.6 °C 27.8 °C

24.2 °C 28.3 °C

temperature

1,500- “

2,000 27.8-29.6 °C

: 28.9-30.2 °C

3,000- i

4,000 27.4—28.9 °C

Rainfall

(mm)

500-

000

Microhabitat

Granite cave

Old building

Granite cave

Old building

Granite wall

Semiever-

evergreen

Wet

evergreen

Acknowledgements.—We thank Chandana Sooriyabandara

(Director General of Department of Wildlife Conserva-

tion), Laxman Peiris (Research Director of Department

of Wildlife Conservation), the research committee, the

field staff of the Department of Wildlife Conservation

(WL/3/2/1/14/12, and WL/3/2/42/18a,b), and the Conser-

vator General of Forests and the staff of Forest Department

(FRC/5, and FRC/6) for granting permission and provid-

ing help during the field surveys; and Nanda Wickramas-

inghe, Sanuja Kasthuriarachchi, Chandrika Munasinghe,

Rasika Dasanayake, Ravindra Wickramanayake, and P.

Gunasiri at NMSL for assisting while examining collec-

tions under their care. Ashan Geeganage (for Fig. 8b), Hat-

angala Medhananda thero, Buddika Madurapperuma (for

the GIS map), Chamara Amarasinghe, Tharaka Kusum-

inda, Hasantha Wijethunga, D.M. Karunarathna, Kawmini

Karunarathna, Rashmini Karunarathna, and Thesanya Ka-

runarathna provided valuable assistance. This work was

mainly supported by Nagao Natural Environment Foun-

dation (2018-20) grant to SK, and United States National

Science Foundation grants DEB 1555968 and EF 1241885

(subaward 13-0632) to AMB. Finally, we would like to

139m

1,600 m

1,592 m

7.020600 80.788639 1,658 m

7.034775 80.783497 1,574 m

§ 2 = fn

= 3 oS S

< A =

-—

ic)

—

‘ae Ee

ANTENITn

aio

OIiNT

SIOATN

nap ora

OTN] co

WmM_WO TT

m~P_Re]e

o;Toy]o

coyTo} co

colraotwo

Ge | oO} co

NIENITOR

NANT mI] oO

as To ITLN

WTO TE”

‘Oo | © | ©

80.746783

81.135569

81.141675

81.114836

81.127831

80.798794

80.773903

Coordinates

Le fe |

6.510536

7.872931

7.851358

7.860200

7.850547

7.033558

7.028314

District

Ratna-

pura

Polon-

Wet zone

Nuwara-

Bioclimatic

Intermedi-

ate zone

C. kotagamai

dissanayakai

C. kawminiae

Table 10. Distribution and ecological data of the three new Cnemaspis species from Sri Lanka. Abbreviations: m — meters; ha — hectares; mm — millimeters; Lux — light intensity; CR — Criti-

cally Endangered.

Amphib. Reptile Conserv. 348 December 2019 | Volume 13 | Number 2 | e216

Karunarathna et al.

thank Kelum Manamendra-Arachchi, Thasun Amarasing-

he, Craig Hassapakis and Michael Grieneisen for various

support, and anonymous reviewers for their constructive

criticism of an earlier draft that helped to significantly im-

prove this paper.

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lost endemic species of Sri Lanka. Oryx 51: 619-626.

December 2019 | Volume 13 | Number 2 | e216

Three new species of Cnemaspis from Sri Lanka

Karunarathna S, Bauer A, de Silva A, Surasinghe T, So-

maratna L, Madawala M, Gabadage D, Botejue M,

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Nilgala Savannah forest, Uva Province of Sri Lanka.

Zootaxa 4545: 389-407.

Karunarathna S, Poyarkov NA, de Silva A, Madawala

M, Boteyue M, Gorin VA, Surasinghe T, Gabadage

D, Ukuwela KDB, Bauer AM. 2019b. Integrative tax-

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genus Cnemaspis Strauch, 1887 (Reptilia: Squamata:

Gekkonidae) from geographically isolated hill forests

in Sri Lanka. Vertebrate Zoology 64: 247-298.

Karunarathna S, Ukuwela K. 2019. A new species of

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logenetics & Evolution 94: 537-547.

Suranjan Karunarathna began his scientific exploration of biodiversity with the Young Zoologists’

Association of Sri Lanka (YZA) in early 2000, and led the society in 2007 as the President. Suranjan

earned his Masters of Environmental Management from University of Colombo, Sri Lanka, in 2017.

As a wildlife researcher, he studies herpetofaunal ecology and taxonomy, and also promotes science-

based conservation awareness on the importance of biodiversity and its conservation among the Sri

Lankan community. Suranjan is an active member of several specialist groups of IUCN/SSC, and

has served as an expert committee member of the IUCN Global and National Red List development

December 2019 | Volume 13 | Number 2 | e216

Amphib. Reptile Conserv.

Karunarathna et al.

Anslem de Silva M.Sc., D.Sc. (University of Peradeniya, Sri Lanka) started keeping reptiles

at the early age of seven, and he has taught herpetology at the Rajarata University of Sri Lanka

and mentored final-year veterinary students at University of Peradeniya. Anselm has conducted

herpetofaunal surveys in most of the important ecosystems in the country, and has published more

than 400 papers, of which nearly 60 are books or book chapters. Anslem had done yeoman service

to the country and the region for more than 50 years. He is the Regional Chairman of the Crocodile

Specialist Group for South Asia and Iran, Co-Chair of the Amphibian Specialist Group IUCN/SSC

Sri Lanka. Anslem received the IUCN/SSC Sir Peter Scott Award for Conservation Merit in October

2019 — the first Sri Lankan to receive this prestigious award.

Madhava Botejue has been engaged in research on the biodiversity, ecology, distribution,

behavior, taxonomy, and conservation of Sri Lankan fauna for the past 14 years, with a main focus

on herpetofauna, avifauna, and mammals. Madhava has contributed to environmental protection

through many community-based awareness programs on the importance of biodiversity and its

conservation. He earned his B.Sc. degree in Natural Sciences from The Open University of Sri

Lanka in 2009. Currently, he serves as an Environmental Officer at the Central Environmental

Authority, Sri Lanka, a member of IUCN/SSC Crocodile Specialist Group, and an expert committee

member of IUCN Global and National Red List development programs.

Dinesh Gabadage is a field biologist who began his wildlife interests in 1990 as a member of the

Young Zoologists Association of Sri Lanka (YZA), and also in 1994 as a member of the Wildlife

Heritage Trust of Sri Lanka (WHT). Dinesh is a dedicated researcher studying the biodiversity

ecology, distribution, behavior, and taxonomy of herpetofauna, avifauna, and mammals in Sri

Lanka; and he has conducted many community-based programs promoting wildlife conservation.

He is also an expert committee member in the IUCN Global and National Red List development

programs, and earned his Diplomas in Palaeo-biodiversity and Zooarchaeology from University of

Kelaniya, Sri Lanka.

Lankani Somaratne, B.Sc. (University of Colombo, Sri Lanka), is a zoologist who started her

career as an Assistant Director in the Zoology Division of the Department of National Museums five

years ago. Lankani has engaged in re-cataloging and updating of the avifaunal, skink, amphibian,

and ichthyological collections at the National Museum for the past five years. She has contributed to

enhancing the knowledge on the museological aspects of Natural Specimen conservation for different

communities. She is a member of the International Community of Museums (ICOM), representing

Sri Lanka. Apart from zoological conservation, she is currently working on conservation project at

the Dutch Museum, Sri Lanka.

Angelo Hettige began his interest in wildlife from a very young age. His interests began to grow

as a member of the Young Zoologists Association of Sri Lanka (YZA) since the early 2000s, from

the junior group continuing up to the senior group. Angelo has contributed to conservation through

community awareness programs on the importance of reptiles and their conservation, and through

numerous snake rescues. Currently, he is working in the snake anti-venom research project at the

University of Peradeniya, Sri Lanka, and he wishes to continue his career studying herpetofauna

and its conservation.

Nimantha Aberathna is a naturalist who began his career and wildlife interests in 2004 as a

naturalist, and as a member of the Youth Exploration Society of Sri Lanka (YES) in 2009. He served

as the President of the Research and Education Committee during 2015-2017. Nimantha holds a

certificate of Wildlife Conservation and Management from the Open University of Sri Lanka. As

a wildlife researcher, he is studying ichthyofauna and orchid ecology and taxonomy. He is also

engaged in a captive breeding program for threatened species, and has been involved in many snake

rescue events. Nimantha worked as a venom extractor for the snake anti-venom research project at

the University of Peradeniya, Sri Lanka.

Majintha Madawala is a naturalist who began his career and wildlife interests in 1995 as a member

of the Young Zoologists Association of Sri Lanka (YZA), and holds a Diploma in Biodiversity

Management and Conservation from the University of Colombo, Sri Lanka. As a conservationist

and a naturalist, he is engaged in numerous habitat restoration, snake rescue programs, and

biodiversity research projects in Sri Lanka. Currently, Majintha is engaged in herpetofaunal research

with the Victorian Herpetological Society in Australia. He is also an active member of the IUCN/

SSC Crocodile Specialist Group and IUCN Global and National Red List development programs.

351 December 2019 | Volume 13 | Number 2 | e216

Amphib. Reptile Conserv.

Three new species of Cnemaspis from Sri Lanka

Gayan Edirisinghe began his studies on wildlife in 2000 when he became a member of the

Young Zoologists Association of Sri Lanka (YZA), which laid a strong foundation for his interest

in mammals. Gayan initiated his research career in 2005 with a study on small mammals. For

the past ten years, he has been involved in many research projects on Sri Lankan fauna, mainly

focusing on the diversity, distribution, ecology, behavior, and conservation of chiropterans. He has

conducted awareness programs to educate the community on the importance of biodiversity and its

conservation, and earned his Diplomas in Palaeo-biodiversity and Zoo-archaeology from University

of Kelaniya, Sri Lanka.

Nirmala Perera has been a member of Young Zoologists’ Association (YZA) since 1999, and he

has conducted several awareness programs on biodiversity conservation through many different

levels of the society. Nirmala also served as the secretary of the action committee of YZA, and

he has worked actively on several environmental issues raised in Sri Lanka. He holds a Diploma

in Biodiversity Management from the University of Colombo and worked as the snake biologist

in the snake venom research project in the Faculty of Medicine, University of Colombo. He also

worked as a Project Coordinator at Udawalawe Human-Elephant Conflict Program of the Born Free

Foundation, Sri Lanka (2011-2014).

Sulakshana Wickramaarachchi is a hardware and software engineer by profession, but began his

studies on wildlife conservation in 2006 as a member of the Young Zoologists’ Association (YZA),

and later served as a committee member of the Research Committee and as the Treasurer during

2011-2012. He is also engaged in captive breeding programs for threatened species and many

snakes rescue missions. He also conducts awareness programs to promote the importance of snake

fauna and its conservation among the Sri Lankan community. Also he worked as an assistant venom

extractor of the snake venom research project in the Faculty of Medicine, University of Colombo.

Thilina Surasinghe is an Assistant Professor in the Department of Biological Sciences in

Bridgewater State University, Mssachusetts, USA, and obtained his Ph.D. in Wildlife Biology

at Clemson University, South Carolina, USA. Thilina is an ecologist; his academic training

encompasses different aspects of biology, ecology, environmental sciences, and natural resources

management. He is experienced in teaching undergraduates in biology, environmental sciences, and

social sciences; and he takes part in projects on landscape-scale biodiversity assessments, Red List

assessments, conservation planning, GIS based threat and GAP analyses, and EPA protocols.

Niranjan Karunarathna is a naturalist who loves traveling, camping, and hiking. He has been a

member of Young Zoologists’ Association (YZA) since 2006, has participated in many herpetological

research projects and also has ongoing funded projects. Niranjan is also conducting wildlife

photography, biodiversity conservation, and educational programs for the Sri Lankan community.

Mendis Wickramasinghe founded the Herpetological Foundation of Sri Lanka (HFS), to further

pursue independent research on the herpetofauna of Sri Lanka, while providing a platform for young

herpetologists to initiate research. With nearly 25 years of field research experience on the herpetofauna

of Sri Lanka, his work has focused on taxonomic identification and biodiversity assessments of

amphibians and reptiles, in an effort to increase awareness on the importance of conserving their

habitats in Sri Lanka. As a result, he has been able to discover and describe 29 new species of

amphibians and reptiles, and participated in the re-discovery of three “extinct” amphibian species.

Kanishka D.B. Ukuwela is currently a Senior Lecturer in Zoology at the Rajarata University of Sri

Lanka. He holds a B.Sc. (Hons.) degree in Zoology from the University of Peradeniya, Sri Lanka

and a Ph.D. in Evolutionary Biology from the University of Adelaide, Australia. His current research

focuses on the origins, evolution, systematics, and conservation of the South Asian herpetofauna.

Aaron Bauer grew up collecting reptiles and amphibians in his native New York. He is the Gerald M.

Lemole Endowed Professor of Integrative Biology at Villanova University in Pennsylvania, USA,

and has been studying reptiles, especially geckos, for more than 35 years. Aaron has worked widely

in Sri Lanka, India, southern Africa, Australia, and the South Pacific; and has described nearly 200

species of reptiles and written more than 750 publications. He is a former Secretary General of the

World Congress of Herpetology, President of the Society for the Study of Amphibians and Reptiles,

President of the Herpetologists’ League, and Chairman of the Herpetological Association of Africa.

352 December 2019 | Volume 13 | Number 2 | e216

Karunarathna et al.

Appendix 1.

Comparative Cnemaspis materials examined from Sri Lanka

Cnemaspis alwisi: NMSL 2004.09.01 (holotype), NMSL 2004.09.02 (paratype), NMSL 2004.09.03 (paratype), WHT 5918, WHT

6518, WHT 6519, WHT 7336, WHT 7337, WHT 7338, WHT 7343, WHT 7344, WHT 7345, WHT 7346.

C. anslemi: NMSL 2019.14.01 (holotype), NMSL 2019.14.02 (paratype), NMSL 2019.14.03 (paratype).

C. amith. BMNH 63.3.19.1066A (holotype), BMNH 63.3.19.1066B (paratype), BMNH 63.3.19.1066C (paratype).

C. butewai: NMSL 2019.07.01 (holotype), NMSL 2019.07.02 (paratype), NMSL 2019.07.03 (paratype).

C. gemunu: AMB 7495 (holotype), AMB 7507 (paratype?), WHT 7221, WHT 7347, WHT 7348, NMSL 2006.11.01, NMSL

2006.11.02, NMSL 2006.11.03, NMSL 2006.11.04.

C. godagedarai: NMSL 2019.09.01 (holotype), NMSL 2019.16.01 (paratype), NMSL 2019.16.02 (paratype).

C. gotaimbarai: NMSL 2019.04.01 (holotype), NMSL 2019.04.02 (paratype), NMSL 2019.04.03 (paratype).

C. hitihami: NMSL 2019.06.01 (holotype), NMSL 2019.06.02 (paratype), NMSL 2019.06.03 (paratype).

C. ingerorum: WHT 7332 (holotype), WHT 7330 (paratype), WHT 7331 (paratype).

C. kallima: WHT 7245 (holotype), WHT 7222 (paratype), WHT 7227 (paratype), WHT 7228 (paratype), WHT 7229 (paratype),

WHT 7230 (paratype), WHT 7239 (paratype), WHT 7249 (paratype), WHT 7251 (paratype), WHT 7252 (paratype), WHT 7253

(paratype), WHT 7254 (paratype), WHT 7255 (paratype).

C. kandambyi: WHT 9466 (holotype), WHT 9467 (paratype).

C. kandiana. BMNH 53.4.1.1 (lectotype), BMNH 80.2.2.119A (paralectotype), BMNH 80.2.2.119B (paralectotype), BMNH

80.2.2.119C (paralectotype), WHT 7212, WHT 7213, WHT 7267, WHT 7305, WHT 7307, WHT 7308, WHT 7310, WHT 7313,

WHT 7319, WHT 7322.

C. kivulegedarai: NMSL 2019.08.01 (holotype), NMSL 2019.08.02 (paratype), NMSL 2019.08.03 (paratype).

C. kohukumburai: NMSL 2019.05.01 (holotype), NMSL 2019.05.02 (paratype), NMSL 2019.05.03 (paratype).

C. kumarasinghei: NMSL 2006.13.01 (holotype), NMSL 2006.13.02 (paratype).

C. latha: WHT 7214 (holotype).

C. menikay: WHT 7219 (holotype), WHT 7218 (paratype), WHT 7349 (paratype).

C. molligodai: NMSL 2006.14.01 (holotype), NMSL 2006.14.02 (paratype), NMSL 2006.14.03 (paratype), NMSL 2006.14.04

(paratype), NMSL 2006.14.05 (paratype).

C. nandimithrai: NMSL 2019.01.01 (holotype), NMSL 2019.01.02 (paratype), NMSL 2019.01.03 (paratype).

C. nilgala: NMSL 2018.07.01 (holotype), NMSL 2018.06.01 (paratype), NMSL 2018.06.02 (paratype), NMSL 2018.06.03 (para-

type).

C. pava. WHT 7286 (holotype), WHT 7281 (paratype), WHT 7282 (paratype), WHT 7283 (paratype), WHT 7285 (paratype), WHT

7288 (paratype), WHT 7289 (paratype), WHT 7290 (paratype), WHT 7291 (paratype), WHT 7292 (paratype), WHT 7293 (para-

type), WHT 7294 (paratype), WHT 7295 (paratype), WHT 7296 (paratype), WHT 7297 (paratype), WHT 7298 (paratype), WHT

7299 (paratype), WHT 7300 (paratype), WHT 7301 (paratype), WHT 7302 (paratype).

Amphib. Reptile Conserv. 353 December 2019 | Volume 13 | Number 2 | e216

Three new species of Cnemaspis from Sri Lanka

C. phillipsi. WHT 7248 (holotype), WHT 7236 (paratype), WHT 7237 (paratype), WHT 7238 (paratype).

C. podihuna: BMNH 1946.8.1.20 (holotype), NMSL 2006.10.02, NMSL 2006.10.03, NMSL 2006.10.04.

C. pulchra. WHT 7023 (holotype), WHT 1573a (paratype), WHT 7011 (paratype), WHT 7021 (paratype), WHT 7022 (paratype).

C. punctata: WHT 7256 (holotype), WHT 7223 (paratype), WHT 7226 (paratype), WHT 7243 (paratype), WHT 7244 (paratype).

C. rajakarunai: NMSL 2016.07.01 (holotype), DWC 2016.05.01 (paratype), DWC 2016.05.02 (paratype).

C. rammalensis: NMSL 2013.25.01 (holotype), DWC 2013.05.001.

C. retigalensis: NMSL 2006.12.01 (holotype), NMSL 2006.12.02 (paratype), NMSL 2006.12.03 (paratype), NMSL 2006.12.04

(paratype).

C. samanalensis: NMSL 2006.15.01 (holotype), NMSL 2006.15.02 (paratype), NMSL 2006.15.03 (paratype), NMSL 2006.15.04

(paratype), NMSL 2006.15.05 (paratype).

C. scalpensis: NMSL 2004.01.01 (neotype), NMSL 2004.02.01, NMSL 2004.03.01, NMSL 2004.04.01, WHT 7265, WHT 7268,

WHT 7269, WHT 7274, WHT 7275, WHT 7276, WHT 7320.

C. silvula: WHT 7208 (holotype), WHT 7206 (paratype), WHT 7207 (paratype), WHT 7209 (paratype), WHT 7210 (paratype),

WHT 7216 (paratype), WHT 7217 (paratype), WHT 7018, WHT 7027, WHT 7202, WHT 7203, WHT 7220, WHT 7354, WHT

7333.

C. tropidogater: BMNH 71.12.14.49 (lectotype), NMSL 5152, NMSL 5151, NMSL 5159, NMSL 5157, NMSL 5970, NMSL 5974.

C. upendrai: WHT 7189 (holotype), WHT 7184 (paratype), WHT 7187 (paratype), WHT 7188 (paratype), WHT 7181 (paratype),

WHT 7182 (paratype), WHT 7183 (paratype), WHT 7185 (paratype), WHT 7190 (paratype), WHT 7191 (paratype), WHT 7192

(paratype), WHT 7193 (paratype), WHT 7194 (paratype), WHT 7195 (paratype), WHT 7196 (paratype), WHT 7197 (paratype),

WHT 7260 (paratype).

Amphib. Reptile Conserv. 354 December 2019 | Volume 13 | Number 2 | e216

Official journal website:

amphibian-reptile-conservation.org

Editorial

Amphibian & Reptile Conservation

13(2): xxx—xxxi (€217).

Manuscript reviewers for Amphibian & Reptile Conservation (2019)

Citation: Hassapakis CL, Grieneisen ML, Conradie W. 2019. Manuscript reviewers for Amphibian & Reptile Conservation (2019). Amphibian & Reptile

Conservation 13(2): xxx—xxxi (e217).

tion 4.0 International (CC BY 4.0): https://creativecommons.org/licenses/by/4.0/], which permits unrestricted use, distribution, and reproduction in any

medium, provided the original author and source are credited. The official and authorized publication credit sources, which will be duly enforced, are

as follows: official journal title Amphibian & Reptile Conservation; official journal website: amphibian-reptile-conservation.org.

Received: 30 December 2019; Accepted: 30 December 2019; Published: 31 December 2019

In 2019, ARC published 49 numbered items, including

45 peer-reviewed articles, 3 book reviews, and a collec-

tion of personal tributes to the late Bill Branch. We are

very grateful to the peer-reviewers listed below for vol-

unteering their time and expertise to help us determine

which of the submitted manuscripts that passed our ini-

tial screening merited publication, and also for providing

suggestions that improved the analysis, interpretation,

and presentation of the data and ideas in each article. The

following individuals have provided either manuscript

reviews during 2019 or prior reviews of manuscripts

that were published in 2019 (Volume 13). The names of

those who reviewed more than one manuscript are given

in bold. We look forward to the continuing generous gift

of time from the reviewers who will help us maintain the

high quality of articles published in ARC in the future.

The primary affiliations of these reviewers include

40 countries: Argentina, Bangladesh, Brazil, Bulgaria,

Canada, China, Colombia, Costa Rica, Czech Republic,

Ecuador, England, Germany, Greece, Guatemala, Hon-

duras, India, Indonesia, Israel, Italy, Japan, Malaysia,

Mexico, Netherlands, New Zealand, Pakistan, Panama,

Peru, Portugal, Romania, Russia, Scotland, Serbia, South

Africa, Spain, Switzerland, Turkey, Ukraine, USA, Ven-

ezuela, and Vietnam.

Stephenson Hallison Formiga Abrantes (Brazil)

Alberto Abreu Grobois (Mexico)

Manuel E. Acevedo (Guatemala)

Bahadir Akman (Turkey)

Muhammed Muassir Ali (Turkey)

Matthew C. Allender (USA)

Ronn Altig (USA)

Steven Anderson (USA)

Manuel Aranda (Mexico)

Adrian Armstrong (South Africa)

Alejandro Arteaga (Ecuador)

César Luis Barrio Amordos (Costa Rica)

Jude Brooke (USA)

Luis Daniel Avila Cabadilla (Mexico)

Ernst Baard (South Africa)

James Barnett (Canada)

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16.

Correspondence. arc.publisher@gmail.com

Amphib. Reptile Conserv.

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. Jelka Crnobrnyja-Isailovic (Serbia)

. Fabio German Cupul Magafia (Mexico)

. Indraneil Das (Malaysia)

. Carlos Delgado-Trejo (Mexico)

. Jesse Delia (USA)

. Leida Dos Santos (New Zealand)

. Lourdes Echevarria (Peru)

. Eduardo G. Etchepare (Argentina)

. Vanda Lucia Ferreira (Brazil)

. Frederico Gustavo R. Franga (Brazil)

. S.R. Ganesh (India)

. Ana Bertha Gatica Colima (Mexico)

. Victor Hugo Gonzalez Sanchez (Mexico)

. Irene Goyenechea (Mexico)

. Eva Gracia (Spain)

. Monica Guerra (Ecuador)

. Amanda Guthrie (USA)

. Vicente Guzman Hernandez (Mexico)

. Md. Kamrul Hasan (Bangladesh)

Abel Antonio Batista Rodriguez (Panama)

Aaron Bauer (USA)

Chris Beirne (USA)

David Blackburn (USA)

Sergé Bogaerts (Netherlands)

Wolfgang Bohme (Germany)

Leo J. Borkin (Russia)

Donald Brown (USA)

Marius Burger (South Africa)

Patricia A. Burrowes (USA)

Henrique Caldeira Costa (Brazil)

Jonathan Campbell (USA)

Carlos Eduardo Costa de Campos (Brazil)

Onur Candan (Turkey)

Fernando Castro Herrera (Colombia)

Luis Ceriaco (Portugal)

Gerardo Chaves (Costa Rica)

Marcio Chaves (Brazil)

Wei Chen (China)

Basundhara Chettri (India)

Kerim Cicek (Turkey)

Ibrahim Hakki Cigerci (Turkey)

Andrea Costa (Italy)

December 2019 | Volume 13 | Number 2 | e217

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Md. Mosharrof Hossain (Bangladesh)

Michael Itgen (USA)

Ulrich Joger (Germany)

Gregor Jongsma (USA)

Mert Karis (Turkey)

M. Monirul H. Khan (Bangladesh)

Akin Kirag (Turkey)

David Kizirian (USA)

Panagiotis Kornilios (Spain)

Brian Kubicki (Costa Rica)

Mirza Kusrini (Indonesia)

Elizabeth Labastida Estrada (Mexico)

James Labisco (England)

William W. Lamar (USA)

Sebastian Lotzkat (Germany)

Vinh Quang Luu (Vietnam)

Fabio Maffei (Brazil)

Stephen Mahony (England)

Anita Malhotra (England)

Ricardo Luria Manzano (Mexico)

Vicente Mata-Silva (USA)

Jose Antonio Mateo Miras (Spain)

Daniel Medina (Brazil)

Shai Meiri (Israel)

+Joseph Mitchell (USA)

Ivelin Mollov (Bulgaria)

Irina T. Morales Castafio (Colombia)

John Mulder (Netherlands)

Edgar E. Neri-Castro (Mexico)

Noorun Nisa (Pakistan)

Cristiano Campos Nogueira (Brazil)

Kei Okamoto (Japan)

Dario Ottonello (Italy)

Andrés Leonardo Ovalle (Colombia)

Cansu Ozbayer (Turkey)

Panayiotis Pafilis (Greece)

Davi Lima Pantoja Leite (Brazil)

Theodore J. Papenfuss (USA)

Gabriela Parra-Olea (Mexico)

Johannes Penner (Germany)

Darren Pietersen (South Africa)

100. Daniel Pincheira-Donoso (England)

101.Juan M Pleguezuelos (Spain)

102. Louis Porras (USA)

Craig Hassapakis, Founder, Publisher, Co-editor

Michael L. Grieneisen, Co-editor

Werner Conradie, Africa Regional Editor

Amphib. Reptile Conserv.

Hassapakis et al.

103.Cynthia P.A. Prado (Brazil)

104. Alex Pyron (USA)

105.Gustavo Ernesto Quintero Diaz (Mexico)

106.Aurelio Ramirez-Bautista (Mexico)

107. Kelsey Reider (USA)

108.Roberto Luna Reyes (Mexico)

109. Mark-Oliver R6del (Germany)

110. Liliana Patricia Saboya Acosta (Colombia)

111. Xavier Santos Santiro (Portugal)

112. Ulrich Schepp (Germany)

113.Gustavo J. Scrocchi (Argentina)

114.Celsi Sefiaris (Venezuela)

115. Shirley Jennifer Serrano Rojas (Scotland)

116. Neftali Sillero (Portugal)

117.Roberto Sindaco (Italy)

118. Ulrich Sinsch (Germany)

119. Brian Slough (Canada)

120. Jiti Smid (Czech Republic)

121.Alexandru Strugariu (Romania)

122. Bryan Stuart (USA)

123.Ben Tapley (England)

124. Krystal Tolley (South Africa)

125.Boris Tuntyev (Russia)

126.Sylvain Ursenbacher (Switzerland)

127.Anna Vassilieva (Russia)

128.Deepak Veerappan (India)

129. Luke Verburgt (South Africa)

130.S.P. Vijayakumar (USA)

131.Jaime Villacampa (Spain)

132.Gernot Vogel (Germany)

133.Rudolf von May (USA)

134. Philipp Wagner (South Africa)

135.Kai Wang (USA)

136. Yanping Wang (China)

137.Steven Whitfield (USA)

138. Larry David Wilson (Honduras)

139.Deniz Yalcinkaya (Turkey)

140.Mehmet Zulfti Yildiz (Turkey)

141.Tonglei Yu (China)

142. Robert T. Zappalorti (USA)

143.Bao Zhang (China)

144.Guangzhou Zhou (China)

145.Oleksandr Zinenko (Ukraine)

146.Marco Alberto Luca Zuffi (Italy)

December 2019 | Volume 13 | Number 2 | e217