Official journal website:

amphibian-reptile-conservation.org

Amphibian & Reptile Conservation

14(1) [General Section]: 1-9 (e218).

The ecology, distribution, status, threats, and conservation

of the Common Water Monitor (Varanus salvator) in the

Dhaleswari River of Assam, India

‘*Muhammed Khairujjaman Mazumder, ?Amir Sohail Choudhury, 7Rofik Ahmed Barbhuiya,

3Himabrata Chakravarty, and *Badruzzaman Barbhuiya

'Department of Zoology, Dhemaji College, Dhemaji, 787057, Assam, INDIA *Department of Ecology and Environmental Science, Assam University,

Silchar 788011, Assam, INDIA *Department of Zoology, Srikishan Sarda College, Hailakandi 788151, Assam, INDIA *District Level Laboratory,

Public Health Engineering Department, Hailakandi 788155, Assam, INDIA

Abstract.—The Common Water Monitor, Varanus salvator (Laurenti, 1768), is a large monitor lizard distributed

in southern and south-east Asia, including India which remains closely associated with water bodies, such

as rivers and lakes. Although IUCN considers it to be ‘Least Concern,’ the Common Monitor Lizard faces

several threats throughout its global distribution range, and the status of the species is decreasing rapidly. The

Dhaleswari River of Assam (India) is one of the most important abodes of this species, where it is locally known

by the names ‘/rong’ and ‘Shanda.’ Geographically, the Dhaleswari River is located in southern Assam (India),

which falls within the Indo-Burma Biodiversity hotspot area. Unfortunately, most of the wildlife of southern

Assam (India) are poorly studied, and this varanid is one of the most ignored species of the region. The present

study was conducted along the Dhaleswari River, Assam, India, to elucidate the distribution, status, ecology,

threats, and conservation of the Common Water Monitor, and is the first report on this species from this river.

The results show that the Dhaleswari River still serves as a habitat of the species, with a viable population.

Further, the species was found to prefer smaller rivers with clayed soil and bushes, and it faces major threats

from habitat destruction, hunting for flesh and oil, and conflicts with humans. Based on our observations, we

discuss recommendations for the conservation of this large varanid.

Keywords. Asia, Barak Valley, Indo-Burma Biodiversity hotspot, Reptilia, Sauria, Varanidae

Citation: Mazumder MK, Choudhury AS, Barbhuiya RA, Chakravarty H, Barbhuiya B. 2020. The ecology, distribution, status, threats, and conservation

of the Common Water Monitor (Varanus salvator) in the Dhaleswari River of Assam, India. Amphibian & Reptile Conservation 14(1) [General Section]:

1-9 (e218).

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 March 2018; Accepted: 23 May 2019; Published: 30 January 2020

Introduction

The Common Water Monitor (Varanus salvator) is the

largest monitor lizard of India, and second largest of

the world, after the Komodo Dragon (V. komodoensis).

It has the widest global distribution among all varanids

(Traeholt 1994). The distribution of the species in the

Indian subcontinent was described by Das (1994), Daniel

(1983), and Koch et al. (2013), while Smith (1935)

reported its occurence and distribution in Northeast

India, of which the state of Assam 1s a part. Anderson

(1982) and Auffenberg (1986) reported the occurence

of this species from Assam (India). Among the four

recognized subspecies of the Common Water Monitor,

V. s. macromaculatus occurs in Assam (Traeholt 1994;

Auffenberg 1986). The Common Water Monitor is

generally found associated with water systems including

Correspondence. *khairujjaman1987@gmail.com

Amphib. Reptile Conserv.

rivers and wetlands (Ahmed et al. 2009; Cota et al. 2009).

It is seldom found far from water (Smith 1935), and only

rarely beyond 200 m from water bodies (Cota et al. 2009).

The Common Water Monitor has a wide trophic niche,

and it consumes a variety of prey species including crabs,

fishes, other lizards, snakes, domestic fowl, and eggs of

other animals, in addition to scavenging (Ahmed et al.

2009).

Some of the important publications on the reptiles

of Northeast India include Ahmed et al. (2009), Das

(2008), and Choudhury (1989, 1992, 1993a,b, 1995,

1996a,b, 1998, 2011). However, none of these articles

provide special treatment of either the Common Water

Monitor, or southern Assam. Southern Assam (India),

is also known as the Barak Valley, and comprises the

districts of Cachar, Hailakandi, and Karimganj. It is

a part of the Indo-Burma Biodiversity hotspot, and

January 2020 | Volume 14 | Number 1 | e218

Varanus salvator in Assam, India

92°27' E 92°40°E

Barak river

24°20'N

te]

Legends

P: Panchgram

K: Kanchanpur

B: Bowerghat

M: Monacherra

L: Lala Town

24°38' N

8: Sahabad

Fig. 1. (A) Map of India, highlighting Assam. (B) Map of

Assam, highlighting Hailakandi district. (C) Map of Hailakandi

district showing the two rivers, Dhaleswari River and Katakhal

River, with distribution of the Common Water Monitor

(Varanus salvator). Dots represent sighting locations during

the present survey; current distribution in the Dhaleswari River

is shown in green. Further downstream, despite no present

records, occurrence in the past (1970s—1980s) was reported by

several interviewees (shown in yellow). No reports on present

occurrence in the Katakhal River (shown in red) could be

found. Map by A.S. Choudhury.

harbors a myriad wildlife assemblage (Myers et al. 2000;

Choudhury 1997, 2013; Mazumder 2014), including a

diverse herpetofauna (Ahmed et al. 2009). Two species

of monitor lizard are reported to inhabit the region,

the Bengal Monitor (Varanus bengalensis) [Das 2008]

and the Common Water Monitor (Varanus salvator)

[Ahmed et al. 2009; Whitaker and Whitaker 1980]. Once

common in most of the rivers of Assam, the habitat of the

Common Water Monitor is decreasing rapidly (Ahmed

et al. 2009), although its IUCN status currently remains

Least Concern (Bennet et al. 2010).

Amphib. Reptile Conserv.

The present study was conducted in the Dhaleswari

River of southern Assam (India) to elucidate the

distribution (past and current), status, ecology (behavior

and habitat), threats, and conservation of the Common

Water Monitor. Based on the findings of this study, we

provide recommendations for the conservation of this

species. The present study is particularly important

since some of the majestic mega-fauna of southern

Assam, including Gaur (Bos gaurus), Asiatic Wild Water

Buffalo (Bubalus arnee), Indian One-horned Rhino

(Rhinoceros unicornis), Sumatran Rhino (Dicerorhinus

sumatrensis), Javan Rhino (R. sondaicus), Royal Bengal

Tiger (Panthera tigris), and notably two mega-reptiles:

the Gharial (Gavialis gangeticus) and Marsh Crocodile

(Crocodylus palustris), have been extirpated in the last

century (Choudhury 1997, 1998, 2013, 2016; Singha

2009), while the Ganges River Dolphin (Platanista

gangetica gangetica) is on the brink of extirpation

(Mazumder et al. 2014).

Materials and Methods

Study site. The present study was conducted in the

Dhaleswari River, located in the district of Hailakandi,

in southern Assam of India. The climate is of the tropical

monsoon type, with average elevation of 21 m asl,

average annual precipitation of 2,400—2,800 mm, and

the temperature varies from 37 °C in the summer to 7

°C in the winter (Choudhury and Choudhury 2017).

The Dhaleswari River originates in the Mizo Hills in

the state Mizoram (India), flows through hilly terrains

towards north, and enters the Hailakandi district. Further

downstream, the river bifurcates at Shahabad (Gainja-

Khauri) of Hailakandi (24°28°51.8°N, 92°34738.3”E),

whereby its water is diverted to an artificial channel

now called Katakhal River with a sluice gate, thereby

eliminating the immediate downstream _ section.

However, further downstream, smaller streams join the

river, thereby enhancing its flow discharge, and it finally

drains into the Barak River at Panchgram, Hailakandi

(24°51°54’”N, 92°36°32”E; Fig. 1). The length of the

river from Shahabad to Panchgram is approximately 110

km, with an elevation ranging from 20-33 m. The river is

almost dry with no flow in the winter season, especially

in the upstream half of its length, while significant flow is

attained in the monsoon season (April to October).

The present study was conducted in a stretch of the

river from Shahabad (24°28751.8"N, 92°34’38.3”E) to

Bowerghat (24°38°37.4"N, 92°32713”E), covering an

approximate length of 56 km. The area is inhabited by

Bengali community people who cultivate rice paddies

(between June and November) in the plain areas, and

vegetables (between November and March) in the plains

as well as on the river banks. The river bank has an

abundance of bushes and bamboo groves, as well as large

trees. Brief surveys were also made in the Katakhal River

(the other river of Hailakandi district) from Sahabad to

January 2020 | Volume 14 | Number 1 | e218

Mazumder et al.

Te eek

Fig. 2. Photographs relevant to the habitat and threats of the Common

a Ee Nee , aS

Water Monitor (Varanus salvator) in the Dhaleswari River,

PE? i =o =

Assam, India. (A) Research team interacting with the locals at Rongpur 5, Hailakandi. (B) Habitat of the Common Water Monitor in

the Dhaleswari River, showing bushes and other features on the banks. (C) The sandy bank of the Katakhal River, prone to erosion

and landslides, is the habitat not preferred by the Common Water Monitor. (D) The sluice gate at the mouth of Dhaleswari River at

Shahabad, which prevents water flow into it and diverts the water to the Katakhal River. (E) Encroachment and conversion of the

Dhaleswari River into fisheries by the locals building dikes at Rongpur 2. Photos by A.S. Choudhury.

its confluence with the Barak River at Katakhalmukh

(24°51°21”N, 92°37°27°E; Fig. 1). The Katakhal River

has greater depth, width, flow rate, and flow discharge,

and it has abrupt banks (with sandy soil) which experience

frequent and severe erosion and landslides.

Survey techniques. Preliminary discussions with the

riparian local people and forest officials were conducted

to locate extant populations in the Dhaleswari and

Katakhal rivers of Hailakandi, southern Assam. The

discussions revealed that the Common Water Monitor

Amphib. Reptile Conserv.

does not occur in the Katakhal River, while it continues

to inhabit the Dhaleswari River between Shahabad and

Bowerghat (Fig. 1). Two researchers independently

carried out an on-foot survey simultaneously along

both banks of the Dhaleswari River, covering a length

of 56 km (of the total river length of ~110 km), from

Shahabad to Bowerghat, to conduct focal counts of the

species and the number of nests available in the river

bank. The burrows made by the lizard were considered to

be nests, while others, including crevices and burrows of

other animals, were considered temporary refuges. The

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Varanus salvator in Assam, India

Table 1. Geo-coordinates of locations along the Dhaleswari River, Assam, India, where live specimens of the Common Water

Monitor (Varanus salvator) were sighted during the survey, including the number of individuals sited at each location.

Geo-coordinates No. of individuals

Site No. Location :

Latitude Longitude sighted

1 Shahbad 24°28'°51.8°N 92°34’38.3"E 1

2 Shahabad 1 24°29°31"N 92°34’33”E 1

3 Rongpur 2 24°30°18.4"N 92°34’39.8"E 1

4 Rongpur 5 24°31°31”"N 92°34’ 36"E 1

5 Rongpur 5 24°31°39.7°N 92°35’°00.7"E 2

6 Rongpur 4 24°31°42.6"N 92°34’°57.7°E 2

ei Abdullahpur 1 24°32’08”"N 92°35°23”E 2

8 Tantoo Road Bridge 24°32°22.1"N 92°35°33.2”E 1

9 Dhanipur 24°33’29"N 92°35’48”"E 2

10 Lala Rural College 24°33°32”N 92°36 12°E 2

11 Lalaghat Nala confluence 24°33°36”N 02°35 21°E 3

12 Kaya Khal confluence 24°34’00”"N 92°34’?24”E 2

13 Bhabanipur 24°34°19"N 92°34’54”°E 2

14 Sarbanandapur 24°34°19"N 92°34’51°E 1

15 Behula 24°34’44"N 92°34’03”E 1

16 Aenakhal Tea Factory 24°35’18"N 92°33’04”E 1

17 Aenakhal Market 24°35°55”N 92°32°352°R 1

18 Monacherra (Lakhinagar) 24°36’45"N 92°32’54”E 3

19 Kukinagar 24°37°26"N 92°32’22”E 2

20 Bowerghat 24°38°37.4"N 92°32713”"E 1

diameter of each nest hole was measured and topology

was identified whenever possible. Different types of nests

were recorded, which were verified by local guides. Upon

sighting of a lizard, the Global Positioning System (GPS)

coordinates were recorded using a digital GPS machine

(eTrex 20X, Garmin, China). The surveyors made keen

observations in the trees for possible occurrences of the

varanid, since this species has been reported to climb

trees for basking (Ahmed et al. 2009). Since no specimens

were found in the Dhaleswari River further downstream

of Bowerghat, or in the Katakhal River, those negative

data are not incorporated into the further analysis. The

total time spent in the field survey, for direct observations

and counting of the lizards, was 118 h for each observer.

In addition, potential threats to the monitor lizard and its

habitat were observed during the survey.

Interviews of local people. Interviews of the locals

(n = 30), generally the older folks, were conducted

regarding the occurrence of the lizards, their nesting

places, food, incidences of hunting, and potential

conflicts (Fig. 2A). In addition to the local people of

the area, many others hailing from different localities in

southern Assam were also interviewed to get an idea of

the past and present occurrences of the monitor lizard

in the Hailakandi district, and in the Dhaleswari River

and Katakhal River in particular. The interviews used

colored photographs, pictorial guides and sketches, and

Amphib. Reptile Conserv.

a closed-ended questionnaire. The total time spent on

the interviews was 55 h.

Results

Distribution and Population Status

Past and current distribution. Interactions with local

people, forest personnel, and elderly people of the

villages revealed that in the 1970s, the whole length of

the Dhaleswari River (~110 km) had viable and abundant

populations of the Common Water Monitor. However, the

population declined gradually, and the species currently

inhabits only the section of the river between Shahabad

and Bowerghat (Fig. 1). The species also occurs in the

tributaries or smaller water channels (mu//ahs) joining the

Dhaleswari River in this river section, most importantly

the Lalaghat Nala, Kaya Khal, and others. In addition,

stray individuals have been reported from Kanchanpur

(~10 km downstream of the current range). It 1s often

found near human habitations, crop fields, ponds, and

lakes near the current main distribution range. Neither

interviews nor surveys revealed the occurrence of this

species in the Katakhal River, and we therefore argue that

the species has been extirpated from this river.

Population status. During the study, 32 individuals were

directly sighted at 20 locations in the 56 km of the river

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Mazumder et al.

section surveyed, for a linear density of 0.57 individuals/

km of the river course (Table 1). The total number of

active nests found during the survey was 215, for a linear

density of 3.84 nests/km of the river section. In addition,

the survey found 63 burrows and crevices which were

used by the lizards as refuges.

Ecology

Habitat and nesting ecology. The current habitat of

the monitor in the Dhaleswari River includes areas with

river channel widths of 1—5 m in the winter months, with

no water present. However, in the monsoon season, the

river width may reach up to 30 m, with significant flow

and discharge. The depth of the river in the study area

varies from 0-10 m. The width of the river bank is 10—30

m, and the banks become flooded with the high waters

of the monsoons. The river banks are inclined, and not

vertically abrupt. The approximate distances of human

habitations from the river bank range from 5—50 m. The

river bank is characterized by the presence of bushes,

secondary tree growth, planted woodlands, bamboo

groves, and occasional crop fields (mainly vegetables),

with or without bamboo fencing (Fig. 2B). However, the

lizard is absent in the Katakhal River, which indicates

that it does not prefer a river where flow and discharge

are high, and banks are abrupt and steep with sandy soil

which are prone to erosion (Fig. 2C).

The lizard made nests in the inclined river banks in

areas where anthropogenic pressure is less intensive.

The diameters of the nest entrance holes were 20-25 cm

(23.59 + 1.14 cm; mean + SD; n = 40). The vicinity of

the nests was very clean, due to movement of the lizards,

while the entries of some nests were half-sealed with

soil. The entries of the nests were horizontal, or inclined

downward or upward; however, the tunnel rose upwards

immediately at an approximate distance of ~0.3—0.6 m

from the entrance hole. Several other micro-habitats

were found, which were natural crevices or burrows of

other animals that were used by the monitors, probably

as refuges. In these larger crevices, the lizards generally

occurred in groups of several individuals.

Behavioral ecology. During winter months, the Common

Water Monitor basks in the sun, and has been found

to climb trees for basking, especially in the morning.

During monsoons, the river water level increased and the

nests were flooded. This factor drove the monitors to hide

in the bushes, forage near human settlements, and shelter

in trees. According to the locals (n = 30), the monitor has

been found to feed on fishes, snakes, small lizards, crabs,

and carcasses of other larger animals which were thrown

into the river. There are several poultry farms near the

river banks, as well as slaughter houses near populated

areas, and people often dispose of carcasses and remnants

of slaughtered animals in the rivers. In such places, the

Common Water Monitor was frequently encountered.

Amphib. Reptile Conserv.

Monitors with newborns were observed in the month of

April, suggesting that eggs hatched in the early monsoon

showers which coincided with the nests becoming

submerged by flood waters. Also, the abundance of fish

and other prey organisms were greater in this season,

thereby making it more suitable for the newborns.

Threats

Like any other aquatic or water body associated species,

and the monitors of other habitats, there are several

threats to the Common Water Monitors in the Dhaleswar1

River, which threaten the survival of the species.

Habitat loss and destruction. Most of the rivers of

Assam (India) are under immense human pressure due to

pollution, overfishing, extraction of water for agricultural

and domestic uses, encroachment, and other uses; and

the Dhaleswari River is no exception. The diversion of

the river water to the Katakhal River by the sluice gate

at Shahabad (Fig. 2D) has led to a decline in available

water in the parent stream, and a loss of biodiversity.

This water diversion also invites cultivation on the river

banks. In several places on the banks, people made

fences of bamboo or steel wire, thereby reducing the

available space for the lizards on the banks. However,

due to decreased river flow, erosion, and land-slides,

many sections of the river could not be studied.

The Dhaleswari River is highly polluted due to

dumping of domestic and agricultural wastes, sewage,

carcasses of animals, and other refuse. All the wastes

which are dumped continue to accumulate until the next

monsoon. This leads to deterioration of water quality,

and thereby causes depletion of aquatic diversity and

prey species, although it does not appear to harm the

lizards directly.

Prey species depletion. The Dhaleswari River does not

maintain water flow in the winter months, and it fully

dries up gradually upstream. In addition, people make

dikes in the banks to catch any remaining fish left in the

river, and these dikes are used for human movement as

well. Unfortunately, the upstream section of the river has

been encroached and converted into fisheries, and flow

in certain areas has been diverted resulting in extremely

narrow channels (Fig. 2E). In this situation, prey species,

mainly fishes and those dependent on the fishes, barely

remain in the river. The severe prey depletion is thus

among the most important threats to the Common Water

Monitor in this river.

Hunting. Unfortunately, there were several instances of

hunting of the Common Water Monitor throughout its

range in the river. It is poached for meat, and also for

oil. The tribal community people of the region, mainly

the Manipuri, Khasi, Mizo, Naga, Tea tribes, and others,

hunt the monitor as a protein source. The oil extracted

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Varanus salvator in Assam, India

Fig. 3. A Common Water Monitor (Varanus salvator) killed for

venturing into a human habitation at Rongpur 6, Hailakandi. It

was subsequently buried. Photo by R.A. Barbhuiya.

from the base of the tail of the lizard, called Shandar

tel (‘Oil of the Monitor lizard’ in the local language), 1s

used as a sexual lubricant by man. Although the Bengali

community people, who are the majority in the present

study area, do not consume the lizard themselves, they

catch any individuals that venture into the settlements

and sell them to the tribes. A moderate sized individual

may cost up to 1,000-1,500 INR (~14-21 USD), while

the oil is sold at the rate of 500 INR/L (~7 USD).

Human-monitor lizard conflicts. During the monsoon

months, the preferred sites of nesting or refuge are

submerged, and thus the lizards venture into human

habitations for foraging, mainly in the afternoon hours.

They often prey on domestic poultry or venture into

artificial ponds and prey on fishes. Thus, the local people

who do not consume the meat of the lizard consider them

to be pests and often kill them. Such killing is an age-

old practice. In addition, many people (mainly children)

believe that the lizard may physically harm them, or even

kill them, and that the bite of the lizard 1s poisonous. Thus,

the local people often harm the Common Water Monitor

whenever it occurs within their reach. Despite occasional

attacks on domestic animals and venturing into ponds,

the lizard was not reported by any of the respondents

to actually attack man. In 2011, one individual was

caught from village Kanchanpur, and killed (Saifur

Rahman Laskar, Kanchanpur, pers. comm.). According

Amphib. Reptile Conserv.

to the interviewees, there were about 25 (range 20-30)

incidences of hunting Common Water Monitors in 2014

in the study area. In the month of August 2015, as many

as six cases of such hunting were recorded from the river

section between Bowerghat and Monacherra, a stretch of

~5 km. On 5 September 2015, one lizard ~1.8 m long

(Fig. 3) was killed from Rongpur 6 village for attacking

poultry, and subsequently buried. However, since the

responses of only 30 interviewees were taken, the exact

number of incidences is estimated to be much higher than

is indicated here.

Lack of research and awareness. Unfortunately, the

Common Water Monitors are the least studied of the

large animals in the region. To the best of our knowledge,

there are no reports on the Common Water Monitors from

this region, save for the few records simply reporting

its occurrence, and no mentions in the literature of its

occurrence in Dhaleswari River. Moreover, no specific

awareness campaign has yet been undertaken to educate

the locals, and the lizard remains among the most ignored

wildlife of the region.

Discussion

The freshwater bodies of the world, both lotic and lentic,

are habitats with highly diverse wildlife assemblages

as well as immense anthropogenic pressures, which

threaten the survival of these species (Dudgeon 2000).

Amidst a myriad of threats, the Common Water Monitor

still survives in habitats that are highly disturbed. In the

Dhaleswari River of Hailakandi (Assam, India), there

is still a good population of the species, inhabiting the

upstream stretch of the river which represents about one-

half of the ~110 km total length of the river course. This

varanid used to be very common throughout the region in

the 1970s. However, today the species 1s encountered only

in the 56 km stretch between Bowerghat and Shahabad

(Fig. 1). During the survey, 32 individual Common Water

Monitors were directly sighted in this river section, for a

linear density of 0.57 individuals/km of the river course

(Table 1). However, the linear density of the active nests

was found to be 3.84/km, which suggests that the actual

number of lizards inhabiting the river section is quite a

bit higher than the number of individuals seen.

No present record of the species in the Katakhal

River could be found. Since this river is much larger,

with greater width, depth, flow rate, and flow discharge

compared to the Dhaleswari, it further supports our

assumption that the Common Water Monitor prefers

smaller river habitats (Fig. 2B). In the Katakhal River,

due to high flow velocity and discharge, the banks are

frequently eroded in the monsoons, and thus monitor

nests would be damaged. Since the species appears to use

its nests repeatedly (for more than one year), larger rivers

with sandy banks and prevalent erosion and landslides

(Fig. 2C) would not serve as good habitats, and thus the

January 2020 | Volume 14 | Number 1 | e218

Mazumder et al.

monitors avoid these rivers. This survey reveals that the

nests are made on high grounds, which is consistent with

earlier reports (Biswas and Kar 1980). The direction of

the nests was found to be turned upwards in the present

study area. Biswas and Kar (1980) also mentioned

that the monitor seals the nest hole after laying eggs

by scraping up soil, and we speculated that the half-

sealed nests found in the present study area are probably

similarly sealed and contain eggs that were laid.

During the present field work, some other fauna

from the river section were recorded, which are prey

of either the lizard or its competitors, or which prey on

the eggs and hatchlings of the Common Water Monitor.

Other lizards recorded from the present survey were

Indian Garden Lizard (Calotes versicolor), Tokay

Gecko (Gekko gecko), Bronze Grass Skink (Eutropis

macularia), and Many-lined Grass Skink (Eutropis

multidasciata), and the snakes included Checkered

Keelback (Xenochrophis piscator), Red-necked Keelback

(Rhabdophis subminiatus), Indian Rat Snake (Pitvas

mucosa), Banded Krait (Bungarus fasciatus), Greater

Black Krait (Bungarus niger), King Cobra (Ophiophagus

hannah), Monocled Cobra (Naja kaouthia), Common

Water Snake (Enhydris enhydris), and Indo-Chinese

Rat Snake (Ptvas korros). The amphibians included

Common Asian Toad (Duttaphrynus melanostictus),

Indian Skipping Frog (Euphlyctis cyanophlyctis), Indian

Bull Frog (Hoplobatrachus tigerinus), and Common

Tree Frog (Polypedates teraiensis). The fishes recorded

from the river were Sperata aor, Sperata seenghala,

Channa punctatus, Notopterus notopterus, Clarias

batrachus, Heteropneustes fossilis, Pethia ticto, and

Anabas_ testudineus. The common fishes cultured

by the local people in the ponds and fisheries in the

adjoining areas (which are often prey of the Common

Water Monitor) were Labeo rohita, Clarias gariepinus,

Ctenopharyngodon idella, Cyprinus carpio, Gibelion

catla, Hypophthalmychthys molitrix, and Labeo calbasu.

The water monitor of Dhaleswari River faces immense

threats, which include anthropogenic pressures in terms

of over-fishing, construction of dikes, conversion of the

river into fisheries (Fig. 2E), cultivation of crops, and

fencing of potential cultivable areas on the bank. Further,

the species is frequently hunted as well as persecuted as

a pest (Fig. 3). Thus, habitat destruction, hunting, and

retaliatory killings are the major conservation issues

in the present study area. These threats, in addition to

others, have extirpated the other majestic aquatic mega-

fauna of the river, including the Ganges River Dolphin

Platanista gangetica gangetica (Mazumder et al. 2014).

In spite of all these potential pressures, the Common

Water Monitor could survive well in the river thanks to

certain attributes, most notably its higher adaptability to

human-modified habitats and wider food niche. In fact,

the Common Water Monitor of Dhaleswari River is more

common in town areas, where the dumping of municipal

wastes provides better provisions for this lizard.

Amphib. Reptile Conserv.

Moreover, the many poultry farms on the river banks

deliberately dispose of dead birds near the nests of the

lizards. In some areas where the monitor 1s deliberately

or incidentally provisioned, it is often concentrated.

The Indian Wildlife (Protection) Act (1972) listed the

species under Schedule I, thereby conferring maximum

legal protection. Although the IUCN regards this species to

be stable and considers it “Least Concerned’ (LC), the status

of the population in southern Assam (India) is decreasing.

Thus, the river section between Shahabad and Bowerghat

requires special care, by limiting anthropogenic pressures.

The Assam Fishery Rules (1953) should be enforced, as

they restrict fishing of brood fishes in the spawning season,

and prohibit fishing using specific methods in specific

seasons, including dewatering, which will enhance fish

stock. The construction of dikes, encroachment, and

pollution of the river should be checked, and sewage

should be treated before discharge. Hunting should be

strictly dealt with and the relevant legislation should be

strictly enforced. The people who consume the meat and

use the oil of the lizards, should be motivated to stop doing

so through education and outreach programs. The ecology,

population status, distribution, and threats of the Common

Water Monitor need to be properly evaluated and a specific

long-term conservation action plan should be devised.

Government and non-government organizations should

come forward to work together for the conservation of this

lizard. Above all, we as human beings and the dwellers

of the Indo-Burma Biodiversity hotspot region should

endeavor to save our pristine wildlife assemblage, or

else the Common Water Monitor will be the next large

reptile, after the Gharial and Marsh Crocodile, to become

extirpated from southern Assam, India.

Acknowledgements.—We sincerely acknowledge some

of our local guides and resource people for sharing

their valuable experiences. To name a few, Abdur

Rahim Mazumder (Ujankupa), Saifur Rahman Laskar

(Kanchanpur), Lukan Uddin Laskar (Monacherra),

Hanna Momtaz Begum Mazumder (Lakshminagar), Ali

Akbar Barbhutya (Krishnapur), Faruk Ahmed Barbhuiya

(Shahabad), Zinna Mohd. Badruzzaman Mazumder

(Ujankupa), Riyazul Azhar Laskar (Bowerghat), Imrana

Begam Choudhury (Ujankupa), Badrun Nehar Laskar

(Ujankupa), Yasmin Choudhury (Hailakandi town),

and Rojob Uddin Laskar (Monacherra) deserve special

thanks. Ruhul Amin Ahmed (Mohanpur, Hailakandji),

Jabed Ahmed (Chiporsangan, Hailakandi), Hanif

Mohd. Choudhury (Shahabad), Arif Ahmed Barbhuiya

(Krishnapur), and Mizazur Rohaman Mazumder

(Hailakandi town W/N-IV, Hailakandi) accompanied

us during field visits. We are especially thankful to Dr.

Anwaruddin Choudhury, an eminent Naturalist and

Conservation Biologist, for his valuable suggestions. We

acknowledge the sincere efforts of the three anonymous

reviewers and the handling editor Halli Boman for

suggesting improvements and corrections.

January 2020 | Volume 14 | Number 1 | e218

Varanus salvator in Assam, India

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January 2020 | Volume 14 | Number 1 | e218

Amphib. Reptile Conserv.

Mazumder et al.

Muhammed Khairujjaman Mazumder is currently working as Assistant Professor in the

Department of Zoology, Dhemaji College, Assam, India. Muhammed obtained his Ph.D. from the

Department of Life Science and Bioinformatics, Assam University, Silchar, India, in the subject

of Neurobiology. He has a keen interest in the natural history of Northeast India, particularly

Assam, and has authored several articles in journals of international repute and book chapters on

the natural history of Assam, as well as Northeast India. Muhammed is particularly interested in

mammals, and both aquatic and water-dependent species, their habitats, ecology, and conservation.

Amir Sohail Choudhury developed a keen interest in wildlife in childhood, being motivated by

Dr. Anwaruddin Choudhury (one of the eminent naturalists of Northeast India). Amir completed

post-graduate work in Ecology and Environmental Sciences with a specialization in Wildlife. His

research interests include studying the population dynamics and habitat ecology of the birds and

mammals of Assam, India. He has authored several articles and book chapters on the natural

history of Northeast India, particularly Assam. Further, Amir has documented reptiles from the

southern part of Assam, with several publications to his credit. He is a freelancer, and is very

interested in wildlife photography.

Rofik Ahmed Barbhuiya is a young naturalist who has been involved in the study and

conservation of wildlife, especially primates, elephants, and birds, since his school years. Rofik is

a post-graduate in Ecology and Environmental Science at Assam University, Silchar, India, and is

currently pursuing his Ph.D. from the same department on the behavioral ecology of the Capped

Langur of Assam (India). He has conducted several awareness programs for the conservation

of Hoolock Gibbon, Phayre’s Langur, Capped Langur, and elephants, and has several articles

published in international journals.

Himabrata Chakravarty is currently serving as Associate Professor and Head, Department of

Zoology, Srikishan Sarda College, Hailakandi, Assam, India. Himabrata was awarded a Ph.D. by

Assam University, Silchar, India, and his research interests include the ecology of birds, mammals,

and reptiles. He has authored several articles and book chapters on the wildlife of Assam (India),

and has been engaged in rescue, rehabilitation, and conservation programs for a long time.

Himabrata also has a keen interest in wildlife photography.

Badruzzaman Barbhuiya is currently working as an Assistant Chemist in the District Level

Laboratory, Public Health Engineering Department, Government of Assam, Hailakandi, Assam,

India. Badruzzaman has a Chemistry background, and a keen interest in studying water quality

and habitat ecology of aquatic species in southern Assam. He has experience in the analysis of

water parameters in the region, and the mitigation of issues related to drinking water quality.

With immense concern for the environment and climatic change, he remains involved in various

awareness activities with NGOs.

) January 2020 | Volume 14 | Number 1 | e218

Official journal website:

amphibian-reptile-conservation.org

Amphibian & Reptile Conservation

14(1) [General Section]: 10—21 (e219).

Climatic niche, natural history, and conservation status of the

Porthole Treefrog, Charadrahyla taeniopus (Gunther, 1901)

(Anura: Hylidae) in Mexico

‘Raciel Cruz-Elizalde, *Itzel Magno-Benitez, *Christian Berriozabal-lslas, 7Raul Ortiz-Pulido,

2*Aurelio Ramirez-Bautista, and “Raquel Hernandez-Austria

'Museo de Zoologia “Alfonso L. Herrera”, Departamento de Biologia Evolutiva, Facultad de Ciencias, Universidad Nacional Autonoma de México

(UNAM). A.P. 70-399, Ciudad de México CP 04510, MEXICO *Laboratorio de Ecologia de Poblaciones, Centro de Investigaciones Biologicas,

Instituto de Ciencias Bdsicas e Ingenieria, Universidad Autonoma del Estado de Hidalgo, Km 4.5 carretera Pachuca-Tulancingo, 42184, Mineral

de La Reforma, Hidalgo, MEXICO +Programa Educativo de Ingenieria en Biotecnologia. Universidad Politécnica de Quintana Roo. Av. Arco

Bicentenario, M 11, Lote 1119-33, Sm 255, 77500 Cancun, Quintana Roo, MEXICO ‘Departamento de Zoologia, Instituto de Biologia, Universidad

Nacional Autonoma de México, Apartado Postal 70-153, 04510 Ciudad de México, MEXICO

Abstract.—Amphibian species of the family Hylidae exhibit a high degree of endemism in Mexico. To better

understand ongoing declines of many amphibian populations, especially for endemic species that are

particularly vulnerable to extinction, information on diverse aspects of their biological makeup is required,

including their ecology. This study provides an analysis of the distribution, natural history, feeding habits,

reproduction, morphology, and conservation status of Charadrahyla taeniopus, a species endemic to central

Mexico. The distribution of this species extends along the Sierra Madre Oriental, primarily in cloud forest.

Based on changes in climatic niche, decreases of 14.14% and 37% of its distributional range are predicted to

occur by the years 2050 and 2070, respectively. An examination of the stomach contents from 31 adults and

two juveniles revealed plant materials and arthropods as major parts of their diet. Charadrahyla taeniopus is

sexually dimorphic in size. Females were larger than males, and after correcting for body size, females had

larger jaws than males. Based on guidelines proposed by national legislation (NOM-059), we propose that this

species should continue to be classified as Threatened. Further studies are necessary to classify it in a high

conservation category by international legislation (IUCN) guidelines, due to the high vulnerability indicated

by the Environmental Vulnerability Score, which is caused by an accelerated loss of habitat. Charadrahyla

taeniopus is a good model for analyzing the conservation status of hylid frogs from temperate areas and in

highly transformed environments, as this species exemplifies the conservation status of endemic amphibians

in central Mexico.

Keywords. Amphibians, Central America, cloud forest, diet, morphology, reproduction

Citation: Cruz-Elizalde R, Magno-Benitez |, Berriozabal-Islas C, Ortiz-Pulido R, Ramirez-Bautista A, Hernandez-Austria R. 2020. Climatic niche,

natural history, and conservation status of the Porthole Treefrog, Charadrahyla taeniopus (Gunther, 1901) (Anura: Hylidae) in Mexico. Amphibian &

Reptile Conservation 14(1) [General Section]: 10-21 (e219).

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: 24 October 2018; Accepted: 16 March 2019; Published: 11 February 2020

Introduction

Scientists are concerned about the worldwide decline of

amphibians associated with habitat degradation (Delia

et al. 2013; Wilson et al. 2013), which is caused by a

variety of factors, including shifts in land use, increased

pollution, and the splitting and fragmentation of habitat

(Becker et al. 2007; Ochoa-Ochoa et al. 2009; Cruz-

Elizalde et al. 2015). As a result, many species are

globally threatened (Lips et al. 2004; Ochoa-Ochoa et

al. 2009) by human activities that are destroying habitat,

leading to population decline, extirpation, or even

species extinction (Delia et al. 2013). The creation of

Correspondence. *ramibautistaa@gmail.com

Amphib. Reptile Conserv.

regulations and laws at national and international levels,

and resources such as the Red List of the International

Union for the Conservation of Nature (IUCN), have been

important tools for the conservation and management of

biodiversity (Wilson et al. 2013).

In Mexico, flora and fauna are protected by the NOM-

059, a regulation that provides a way to evaluate the

threat level or conservation status of species through a

nationally-recognized method: the Método de Evaluacion

del Riesgo de Extincion de las Especies Silvestres en

Mexico (method for evaluation of the extinction risk of

wild species, MER; DOF 2010). MER has been used to

evaluate the conservation status of diverse plant taxa,

February 2020 | Volume 14 | Number 1 | e219

Cruz-Elizalde et al.

om, ae re

Fig. 1. Female individual of Charadrahyla taeniopus (A) in a

cloud forest and (B) at Tenango de Doria, Hidalgo, Mexico.

Photos by Uriel Hernandez-Salinas (A) and Raciel Cruz-

Elizalde (B).

but major groups of animals, including many species of

amphibians, have not yet been evaluated (DOF 2010).

Moreover, the lack of knowledge of the distribution and

natural history of amphibian species populations impedes

the proper application and evaluation of the MER. For

example, Lithobates johni (Blair, 1965) is an endemic

species that is considered endangered, and was reported as

extirpated at the type locality in Palictla, San Luis Potosi,

Mexico (DOF 2010; Campos-Rodriguez et al. 2012).

However, Hernandez-Austria et al. (2015) found several

robust populations of the species in the state of Hidalgo.

According to MER, conservation assessments should

be updated to include current data and to focus on such

critical factors as population density, reproductive period,

and natural history (Wilson et al. 2013). One method for

assessing conservation status that takes these factors into

consideration is the Environmental Vulnerability Score

(EVS). It is based on an algorithm proposed by Wilson

et al. (2013) for amphibian species inhabiting Mexico,

and has been accepted for biodiversity and conservation

studies of this group at a variety of spatial scales (Johnson

et al. 2015; Mata-Silva et al. 2015; Lemos-Espinal et al.

2018a,b). Use of this index has enabled better estimates

of the conservation status of amphibian species at

regional (Cruz-Elizalde et al. 2015, 2016) and state levels

Amphib. Reptile Conserv.

(Johnson et al. 2015; Mata-Silva et al. 2015) by utilizing

information on ecological distribution and reproductive

modes (Wilson et al. 2013).

Approximately 99 species of the family Hylidae

occur in Mexico (Wilson et al. 2013; Parra-Olea et al.

2014; Canseco-Marquez et al. 2017; Johnson et al. 2017;

Jiménez-Arcos et al. 2019), and 68 of these are endemic

to the country (Parra-Olea et al. 2014; Caviedes-Solis

et al. 2015; Canseco-Marquez et al. 2017; Johnson et

al. 2017). Many of the endemic species are distributed

in temperate and tropical environments (Flores- Villela

et al. 2010; Delia et al. 2013). The family Hylidae

exhibits a remarkable species richness and diversity

in montane regions (Duellman 2001; Flores-Villela

et al. 2010), which are at high risk because of a shift

in land use from forests to agroecosystems such as

shade coffee plantations and grazing areas (Ochoa-

Ochoa et al. 2009; Santos-Barrera and Urbina-Cardona

2011; Murrieta-Galindo et al. 2013). Furthermore, the

temperate environments of Mexico are expected to be

affected by climate change, leading to a decrease in

species richness and diversity (Lips et al. 2004; Urbina-

Cardona and Flores-Villela 2010).

The genus Charadrahyla (Faivovich et al. 2005) is

composed of ten species (Frost 2019) which inhabit the

highlands of the Sierra Madre Oriental, Sierra Madre

Occidental, and the sierras of Oaxaca and Chiapas

(Duellman 2001; Campbell et al. 2009; Frost 2019).

Little 1s known about their ecology and natural history,

so most of the species are assigned to categories of

high extinction risk by the IUCN. Four of the species,

C. juanitae (Synder, 1972), C. nephila (Mendelson

and Campbell, 1999), C. pinorum (Taylor, 1937), and

C. taeniopus (Gunther, 1901) are in the Vulnerable

category; C. chaneque (Duellman, 1961) is Endangered;

C. altipotens (Duellman, 1968) and C. trux (Adler

and Dennis, 1972) are Critically Endangered; and C.

sakbah Jiménez-Arcos, Calzada-Arciniega, Alfaro-

Juantorena, Vazquez-Reyes, Blair, and Parra-Olea,

2019, C. esperancensis Canseco-Marquez, Ramirez-

Gonzalez, and Gonzalez-Bernal, 2017, and C. tecuani

Campbell, Blancas-Hernandez, and Smith, 2009 have

not been evaluated (IUCN 2019). According to NOM-

059-SEMARNAT-2010, C. altipotens, C. chaneque,

and C. pinorum are in the Subject to Special Protection

(Pr) category; C. juanitae, C. taeniopus, and C. trux are

Threatened (A); and the remaining four have not been

evaluated (DOF 2010).

Charadrahyla taeniopus (Fig. 1A) inhabits un-

disturbed cloud forests (Fig. 1B) and pine-oak forests in

the Sierra Madre Oriental (Duellman 2001; Kaplan and

Heimes 2015), at elevations from 1,100 to 1,200 m in the

states of Hidalgo, Puebla, and Veracruz (Duellman 2001).

To date, knowledge about the potential distribution,

ecology, reproduction, and natural history of this species

is limited (Duellman 2001; Kaplan and Heimes 2015).

The primary goal of this study is to assess changes in

February 2020 | Volume 14 | Number 1 | e219

Charadrahyla taeniopus in Mexico

the current and future (2050 and 2070) climatic niche

of this species in Mexico. Secondary goals include

characterizing the feeding habits, basic reproductive

parameters, morphological variation, and conservation

status of this species. This information can be used to

develop future conservation strategies in environments

with high species numbers and endemism, such as the

cloud forests of the Sierra Madre Oriental (Ponce-Reyes

et al. 2012).

Materials and Methods

Data Collection

Occurrence data for C. taeniopus were obtained from

(i) the databases of the Global Biodiversity Information

Facility (GBIF), the Comision Nacional para el

Conocimiento y Uso de la Biodiversidad (CONABIO),

and HerpNet; (11) records from publications by Duellman

(2001) and Campbell et al. (2009); and (ii1) specimens

from field work (sporadic collecting in the state of

Hidalgo from 2008 to 2016) deposited in the Coleccion

Herpetologica, Centro de Investigaciones Bioldgicas

at the Universidad Autonoma del Estado de Hidalgo

(Appendix 1).

Climatic Niche Modelling

The 37 unique occurrence records of C. taeniopus were

used to generate climatic niche models. For this study,

climate information was obtained from the 19 current

climate layers available in the WorldClim database version

1.4 (Hijmans et al. 2005). These climate layers contain

the averages of meteorological conditions recorded

from North America, with a spatial resolution of 2.5

arc-min. To avoid overrepresentation of environmental

variables, a bivariate correlation analysis was carried

out with the aim of reducing multicollinearity among the

input variables (Merow et al. 2013; Varela et al. 2014).

For variables that were highly correlated (r > 0.7), the

variable was chosen that exhibited the greatest variation

or that represented the greatest biological meaning for

the actual distribution of the species (e.g., temperature

or precipitation). After this procedure, eight climate

variables were retained: annual mean temperature (BIO

1), temperature seasonality (standard deviation x 100,

BIO 4), maximum temperature of the warmest month

(BIO 5), minimum temperature of the coldest month

(BIO 6), annual temperature range (BIO 5—BIO 6, BIO

7), mean temperature of the wettest quarter (BIO 8),

mean temperature of the coldest quarter (BIO 11), and

precipitation in the driest month (BIO 14; Hijmans et al.

2005). Subsequently, MAXENT was used, the selected

climate variables were projected onto a map of Mexico

and climate change scenarios were estimated (Phillips

and Dudik 2008). Each model was replicated 100 times,

the maximum number of repetitions allowed by available

Amphib. Reptile Conserv.

computing power (Dambach and Rodder 2011). Average

models for the present and for the future years 2050 and

2070 were then obtained. Current and future projections

were estimated using the CCSM-GCM model under

the greenhouse concentration scenario RCP8.5, which

represents a pessimistic scenario (RCP8.5 = +8.5 W/m”).

Finally, to assess the impact of climate change on

habitat suitability for the species, the percent change

between current and future potential distribution areas

was estimated. To determine habitat suitability, the

formula % change = [(S,-S,)/S,]* 100% was used, where

S, 1s the total area that the species occupies in the country

according to the current distribution scenario, and S, is

the total area that the species could occupy in the country

under future climate change conditions (Gutiérrez and

Trejo 2014).

Diet

Stomach contents were removed from 31 adults and

two juveniles (Appendix 1), and a stereomicroscope

was used to classify all identifiable organisms to order,

including plant material. The number of prey items in

each stomach was tallied (n), the number of stomachs

with each prey category (i) was determined (F;,), and the

percentage of stomachs with prey category 1 (YF) was

calculated. Also, the number of prey 1ttems belonging to

each prey category (N) and the numerical percentage of

total abundance (%N) represented by each prey category

were determined. Then the volume of each prey item

(mm*) was calculated using the formula for an ellipsoid

(Selby 1965; Duré and Kehr 2004; Duré et al. 2009): V

= 4/3 n (length/2) (width/2)’. The food importance index

(1) was calculated using the formula of Biavati et al.

(2004), which is I = (F%+N%+V%)/3, where F% is the

percentage of occurrence, N% is numerical percentage,

and V% is volumetric percentage. The trophic niche

amplitude was measured with Levin’s standardized index

using the formula B, = ((1/Xp,7)-1)/n-1, where p, is the

proportion of each prey category with respect to the total

number of prey found in each sex, and 7 is the number of

prey categories in the diet of individuals (Hurlbert 1978).

The overlap in dietary composition between sexes was

analyzed with Pianka’s index (1986): O,, = P,P. a

»P’,,. where p, and P, are the numerical proportions

of prey belonging to the 7" category that was used by

organisms (sexes) j and & (Pianka 1986; Gadsden and

Palacios-Orona 1997). Analyses of amplitude and overlap

for males, females, and juveniles were carried out in the

Ecological Methodology v. 6.1.1. program (http://www.

exetersoftware.com [Accessed: 26 January 2016]).

Reproduction

Thirty-one preserved adults (11 females and 20 males;

Appendix 1) were examined to assess reproductive

parameters. Fat bodies and livers from all individuals,

February 2020 | Volume 14 | Number 1 | e219

Cruz-Elizalde et al.

-100 -99 -98 -97 -96 -100 -99 -98

Gulf of Mexico

Low: 0

02550 100 150 200 “SS

02550 100 150 200

aif ee Km

-97 -96 -100 -99 -98 -97 -96

; NN

Gulf of Mexico Gulf of Mexico

+ =

™ 1 ” 1,

Low: 0 | Low:0 le

~™

02550 100 150 200

= Ps see __ =

Fig. 2. Current (A), and future 2050 (B) and 2070 (C) scenarios of the climatic niche of Charadrahyla taeniopus in Sierra Madre

Oriental province (degraded area). The color scale indicates the probability of occupancy for the species.

and egg mass in females or testes mass in males were

weighed to the nearest 0.0001 g using an analytical

balance. For males, the length and width of testes were

measured to the nearest 0.01 mm with a digital caliper,

and testicular volume was calculated using the formula

for an ellipsoid (Selby 1965; Duré and Kehr 2004). In

females, 10% of the total egg mass was excised from the

oviducts, and the eggs contained therein were counted.

The eggs were counted in a Petri dish with water using

a stereomicroscope. Since the data were not normally

distributed, Spearman’s correlation test was used to

evaluate correlations between body size and either egg

mass or testicular mass. A Mann-Whitney U test was

performed to evaluate the differences in mass, volume,

length, and width of the testes.

Morphology

For all adult specimens (11 females and 20 males),

digital calipers were used to measure the following (each

+ 0.1 mm): snout-vent length (SVL), internarial distance

(IND), eye diameter (ED), interorbital diameter (OD),

tympanum diameter (TD), head length (HL), head width

(HW), head height (HH), jaw length (JL), jaw width (JW),

length from forearm to the fourth finger (LFFT), forearm

length (FOL), length of the humerus (LHU), thigh length

(THL), tibia length (TL), and foot length (FL; Watters et

al. 2016). The normality of the data was checked with a

Kolmogorov-Smirnov test. Since the data were normal,

an analysis of covariance (ANCOVA) was conducted to

analyze sexual dimorphism. The function of the ANCOVA

was to eliminate, through linear regressions, the effect of

SVL (covariate) on the dependent variables (IND, ED,

IOD, TD, HL, HW, HH, JL, JW, LFFT, FOL, LHU, THL,

TL, and FL), and to check whether the regression slopes

were different between the sexes (factor; Zar 2009). The

data are presented as mean + | SE.

Amphib. Reptile Conserv.

Conservation Status

Conservation status was summarized by consulting

Mexican regulations (NOM-059-SEMARNAT-2010;

DOF 2010) and the IUCN (2019) Red List, and an

Environmental Vulnerability Score (EVS; Wilson et

al. 2013) was generated. The EVS recognizes three

categories of risk: low (3-9 points), medium (10-13),

and high (14-19). The score is the result of adding points

assigned to features of a species based on three criteria:

(i) extent of the geographic distribution, (11) extent of

ecological distribution (vegetation types used), and (111)

type of reproductive mode (Wilson et al. 2013).

Results

Areas of Occupation and Exchange Rates of Climate

Availability

The distribution of C. taeniopus was restricted to cloud

forests in the central region of the Sierra Madre Oriental

(Fig. 2A—C) in the states of Hidalgo, Puebla, Veracruz,

and the northern portion of Oaxaca, Mexico. The analysis

of habitat occupancy under current conditions and in

the future (to 2050 and 2070), showed a general loss

of climatic niche in much of the range of C. taeniopus.

This loss occurred in temperate areas, where the current

area of occupation (18,262.23 km’) will decrease to

15,678.45 km? by 2050, which represents a habitat

availability decrease of 14.14% (Fig. 2B); and to an area

of 11,032.93 km? by the year 2070, which represents a

habitat availability decrease of 37.08% (Fig. 2C).

Diet

Fourteen taxa were identified in the stomachs, with 11

taxa present in both males and females (Table 1). The

February 2020 | Volume 14 | Number 1 | e219

Charadrahyla taeniopus in Mexico

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February 2020 | Volume 14 | Number 1 | e219

Amphib. Reptile Conserv.

Cruz-Elizalde et al.

Table 2. Mean values + 1 SE, and range in parentheses, of morphological characteristics of adult Charadrahyla taeniopus (SVL =

snout-vent length, IND = internarial distance, ED = eye diameter, IOD = interorbital diameter, TD = tympanum diameter, HL = head

length, HW = head width, HH = head height, JL = jaw length, JW = jaw width, LFFT = length from forearm to the fourth finger,

FOL = forearm length, LHU = length of the humerus, THL = thigh length, TL = tibia length, FL = foot length). Comparisons were

made with ANCOVA, with SVL as the covariate.

Characteristics Female (” = 11) Male (n = 20) ain

F df P

IND 5.58 + 0.24 (3.87-7.08) 4.94 + 0.18 (2.91-6.72) 1.133 1, 28 0.296

ED 6.22 + 0.23 (5.06-7.91) 5.86 + 0.21 (4.86-8.80) 0.065 1, 28 0.800

IOD 7.02 + 0.36 (4.24-8.74) 5.21 + 0.39 (1.57-8.18) 5.396 1, 28 0.027

TD 3.75 + 0.17 (2.64-4.62) 3.29 + 0.26 (1.99-6.85) 0.457 1, 28 0.504

Ai 19.82 + 0.97 (12.60—24.09) 18.46 + 0.38 (15.81—22.91) 0.002 1, 28 0.966

HW 19.99 + 0.70 (13.94—22.51) 16.45 + 0.32 (14.01-19.41) 24.521 1, 28 <0.001

HH 8.14 + 0.36 (5.56-9.19) 6.83 + 0.32 (4.95-9.08) 3.553 1, 28 0.069

JL 17.28 + 0.66 (12.33—20.68) 15.33 + 0.26 (13.57-17.45) 6.433 1, 28 0.017

JW 19.91 + 0.80 (13.05—22.99) 17.22 + 0.31 (15.20—20.87) 19.166 1, 28 <0.001

LFFT 16.83 + 1.00 (12.03—21.05) 15.60 + 0.53 (11.58-20.37) 0.003 1, 28 0.959

FOL 13.78 + 0.65 (9.44-16.91) 13.12 + 0.29 (11.08-15.58) 1.387 1, 28 0.249

LHU 16.38 + 0.84 (10.77—20.80) 16.07 + 0.51 (10.48-19.26) 2.365 1, 28 0.135

THL 31.32 + 1.44 (20.21-38.05) 30.17 + 0.65 (25.19-37.96) 0.927 1, 28 0.344

su, 32.68 + 1.24 (22.02—37.08) 30.68 + 0.54 (27.84—37.26) 0.016 28 0.900

FL 43.06 + 1.77 (30.21—51.14) 37.92 + 2.17 (34.38-49.26) 0.035 1,27 0.853

most important prey categories, according to the values

of food importance for the species and for each sex, were

orthopterans, plant material (leaves), and ants (Table 1).

The overlap in diet between the sexes was high (O,, =

0.822; 63.21—-100%), with males presenting a slightly

higher value (B = 0.526) of diet niche breadth than

females (B = 0.504).

Reproduction

The mean number of eggs was 722 + 277.53 (range 426—

1,138, n= 11). There was no correlation between female

SVL and either number of eggs (7, = 0.09, P = 0.79, n

= 11) or egg mass in females (r, = 0.09, P = 0.79, n =

11). There were no differences among weights, lengths,

widths, or volumes of the testes (P > 0.05 in all cases).

The average weight, length, width, and volume of the

right testis was 0.198 g, 14.78 mm, 6.05 mm, and 305.57

mm*°, respectively; and for the left testis the averages

were 0.204 g, 14.63 mm, 6.0 mm, and 293.25 mm’,

respectively. There was no correlation between SVL and

testicular volume (7, = 0.31, P = 0.17, n = 20), but there

was a positive correlation between SVL and testicular

mass in males (7, = 0.49, P = 0.02, n = 20).

Morphology

Five of the 15 characteristics measured exhibited sexual

dimorphism, with females higher than males in SVL,

Amphib. Reptile Conserv.

IOD, HW, JL, and JW (Table 2). Females (mean SVL

= 63.94 + 2.35 mm; range 45.27-74.31, n = 11) were

larger than males (mean SVL = 59.70 + 1.09 mm, range

52.90-71.05, n= 20; U= 60, P = 0.04).

Conservation Status

Charadrahyla taeniopus is listed in conservation

standards (DOF 2010; IUCN 2019) as being in high

risk categories. According to the Mexican Standard

NOM-059-SEMARNAT-2010 (DOF 2010), the species

is considered to be Threatened. The IUCN Red List of

Threatened Species places the species in the Vulnerable

category, with a distributional area less than 20,000

km? in fragmented environments and with declining

populations (status Blab[ii]; IUCN 2019). In the EVS, it

was classified as a medium environmental vulnerability

species, with a value of 13 points. This EVS category

was calculated from: (1) its distribution in Mexico,

but not exclusive to the type locality (5 points), (1) its

occurrence in two vegetation types (pine-oak and cloud

forest, 7 points), and (111) a reproductive mode with egg

laying in lentic or lotic water bodies (1 point; Wilson et

al. 2013).

Discussion

Conserving native populations of tree frogs at a local scale

requires information on their ecological distribution,

February 2020 | Volume 14 | Number 1 | e219

Charadrahyla taeniopus in Mexico

feeding habits, reproduction, and morphology (Delia et

al. 2013; Toledo et al. 2014). The analyses reported here

suggest that the distribution of C. taeniopus will potentially

decrease during the next 50 years. Microhabitats in

currently occupied habitats (montane environments) are

subject to change because of temperature and moisture

shifts, and also because of changes in vegetation cover

associated with high deforestation rates (Kaplan and

Heimes 2015) and potential climate change, including

shifts in temperature and moisture (Ponce-Reyes et al.

2012). For example, several authors including Pineda

and Halffter (2004), Pineda et al. (2005), and Murrieta-

Galindo et al. (2013), have suggested that the existence

of abundant vegetation and native shrub cover provide

appropriate humidity and temperature conditions for the

permanence of hylid frogs in temperate environments

such as cloud forests. If the abiotic and biotic conditions

change in the forests inhabited by C. taeniopus, this

species could be negatively affected. Loss of climatic

niche in our models is consistent with that reported by

Roxburgh et al. (2004). These authors mentioned that the

expected changes could generate ecological scenarios

that will delimit the overall distribution of arboreal

species from cloud forests (Roxburgh et al. 2004; Pineda

et al. 2005), and therefore could affect their associations

with their environment (Urbina-Cardona and Flores-

Villela 2010; Ponce-Reyes et al. 2012).

In addition to the above considerations, the thermal

tolerances of anurans in high elevation or low temperature

environments can determine the presence and distribution

of their populations (Wells 2007). The hylid frogs are an

example of this, as their limits of distribution are in high

latitude regions such as the arid and semi-arid climates

of northern Mexico (Wiens et al. 2006). This may be the

result of the thermal tolerances that hylid species show in

temperate environments, which are different from those

of species that occur in tropical environments (Navas

2006; Wells 2007). To date, there are no studies of

thermal tolerances or maximum/minimum temperature

limits for C. taeniopus; therefore, it is very difficult to

know the behavior of individuals and/or populations of

this species in their distribution area. Future field studies,

and in situ and laboratory experiments on thermal

preferences are therefore necessary for this species.

They could complement the results obtained in the

potential distribution model of the species, enabling the

analysis of variables that could be interacting to a greater

degree with the biology of the organism, and improving

determinations of the distribution range of the species

(Gross and Price 2000; Wiens et al. 2006).

Ochoa-Ochoa et al. (2009), stated that in addition to

the loss of vegetation, climate change is a determining

factor in the loss of amphibian species in conserved

environments, mainly in sites outside of natural

protected areas (NPAs). The Sierra Madre Oriental

Corridor occupies large areas of cloud forest, a type of

environment that is highly threatened by the effects of

Amphib. Reptile Conserv.

climate change (Ponce-Reyes et al. 2012), and in which

the known distribution of the species is not included in

any NPA (IUCN 2019). This shows the importance of

evaluating the distribution of highly vulnerable hylid

frogs throughout the potential distribution range based

on climatic niche models and climate change scenarios.

The results are worrisome, because despite the fact that

amphibian richness in Mexico is high (Johnson et al.

2015, 2017), more than 50% of the species are listed in

high vulnerability categories by the IUCN (Delia et al.

2013; Caviedes-Solis et al. 2015; IUCN 2019; Johnson

et al. 2017). For example, recent studies have found that

some mountain hylid species have not been recorded

over prolonged periods of time (Delia et al. 2013;

Caviedes-Solis et al. 2015). Due to multiple factors,

such as vegetation loss, pollution, and in particular

climate change, populations of these species tend to

occur in highly vulnerable sites (Lips et al. 2004; Stuart

et al. 2004). Therefore, the species that inhabit this

type of environment (cloud forest, pine-oak) are highly

threatened (Ochoa-Ochoa et al. 2009; Caviedes-Solis et

al. 2015).

Inaddition to habitat fragmentation and climate change,

the presence of the pathogenic fungus Batrachochytrium

dendrobatidis Longcore, Pessier, and Nichols, 1999 (Bd)

has contributed to amphibian population and species

losses in Mexico (Mendoza-Almeralla et al. 2015, 2016)

and other regions of the world (Lips et al. 2003; Fisher et

al. 2009). However, Bd has not been detected thus far in

C. taeniopus (Murrieta-Galindo et al. 2014; Hernandez-

Austria 2017). Therefore, further studies are needed to

examine the potential presence of Bd in C. taeniopus

populations through their distribution area (Hernandez-

Austria 2017).

The lack of information on the natural history of this

species inhibits the development of strategies for its

conservation (Toledo et al. 2014). The data presented

here on diet provide valuable information on the basic

ecology of C. taeniopus. The diet of this species consists

of orthopterans, plant material, and ants, and there is a

high degree of overlap in diet between the sexes. In C.

taeniopus, plant material is the second most important

food item. This is particularly notable since the diet

of most anuran species in Mexico consists primarily

of arthropods (Ramirez-Bautista and Lemos-Espinal

2004; Suazo-Ortufio et al. 2007), and the ingestion of

plant material, such as leaves and flowers, is usually

considered to be accidental (Evans and Lampo 1996).

In the case of C. taeniopus, further studies are necessary

to determine if consumption of plant material (leaves) is

accidental or part of their diet, which would be unusual,

but not unprecedented. For example, some species of

tree frogs, such as Ptychohyla zophodes Campbell and

Duellman, 2000 (Luria-Manzano 2012) and Xenohyla

truncata (Izecksohn, 1959) do consume large quantities

of plant material, and the latter (XY. truncata) has been

reported as entirely omnivorous, consuming fruits, seeds,

February 2020 | Volume 14 | Number 1 | e219

Cruz-Elizalde et al.

and flowers (da Silva and Britto-Pereira 2006).

Egg number and the relative sizes of eggs vary

greatly in amphibians (Vitt and Caldwell 2009), and they

are often related to female body size (Jorgensen 1992;

Hartmann et al. 2010). The data presented here show

that egg number is not related to female body size in C.

taeniopus. This may be due to the fact that its reproductive

period may have a longer duration, and the sample size

obtained from the collections only reflects the behavior

of the females in the first part of the year (March-April),

not in the entire reproductive period. Females with eggs

were found throughout the year, and aggregations of

individuals of both sexes and amplexus were observed

in the field in August. This seasonal variation in the

correlation between egg size and size of females has been

reported for other anuran species such as Leptodactylus

fuscus (Schneider, 1799), L. podicipinus (Cope, 1862),

and Dendropsophus nanus (Boulenger, 1889) [Prado

and Haddad 2005]. Furthermore, testicular mass, but not

testicular volume, increases with larger SVL. These data

suggest that larger males invest more energy in sperm

production to have greater reproductive success (Byrne

et al. 2002).

Most species of frogs (nearly 90%) are sexually

dimorphic, with females being larger than males (Wells

2007), and C. taeniopus is no exception. The larger size

of females compared to males is presumably associated

with the potential to produce more eggs. However, no

correlation was found between SVL of females and egg

number. Another explanation for sexual size dimorphism

could be differences in growth rates (Kupfer 2007), in

which the growth rate of males is faster than that of

females in order to reach sexual maturity at a smaller size

and compete with other males for access to calling sites,

thereby maximizing the number of matings (Kupfer 2007;

Wells 2007). Also, considering the ecological hypothesis

to explain the sexual dimorphism, the larger jaw size in

females compared to the males might indicate a larger

gape in females, which could allow for partitioning of

food resources in terms of prey size (Luria-Manzano

2012). However, additional studies on microhabitat

use, behavior, and reproduction are required before the

ecological significance of the sexual dimorphism in C.

taeniopus can be determined.

Based on the information about climatic niche,

feeding habits, reproduction, and morphology, C.

taeniopus is highly threatened because it is distributed

in environments (i.e., cloud, oak, and pine-oak forests)

that are currently being dismantled by fragmentation and

climate change (Ponce-Reyes et al. 2012). As with other

hylid frogs (Caviedes-Solis et al. 2015), C. taeniopus

could face a rapid rate of population decline, as has

occurred in other species inhabiting the temperate areas

of cloud forest in Oaxaca (Delia et al. 2013; Mata-Silva

et al. 2015), Chiapas (Johnson et al. 2015), and areas of

the Sierra Madre Oriental (Flores-Villela et al. 2010). To

add to the information presented in this study, additional

Amphib. Reptile Conserv.

studies on demography, ecology, physiological tolerances

to temperature, length of the reproductive period, effect of

fragmentation on populations, and population dynamics

of this species should be conducted in order to devise

efficient conservation strategies for C. taeniopus, and

other species of anurans that inhabit the temperate and

tropical montane environments of central and southern

Mexico (Delia et al. 2013; Caviedes-Solis et al. 2015).

Acknowledgments —We thank Abraham Lozano, J.

Daniel Lara Tufifio, Diego Juarez Escamilla, Concepcion

Puga, Uriel Hernandez Salinas, and Luis M. Badillo

Saldafia for their help in the field and laboratory. We

thank three anonymous reviewers for comments that

greatly improved the manuscript. We thank Margaret

Schroeder for her revision of the English. Thanks to

Irene Goyenechea Mayer Goyenechea for providing the

numbers of specimens from Colecci6n Herpetologica

CIB-UAEH. This study was supported by the CONABIO

JMO001, Fomix-CONACyT-191908 Biodiversidad del

Estado de Hidalgo-3a, UAEH-DI-ICBI-BI-SF-008

and ICBIPAI-29 projects. Specimens were collected

under SEMARNAT-SGPA/DGVS/02726/10 and SGPA/

DGVS/11746/13 permits issued by SEMARNAT and

assessed by Mexican environmental protection laws.

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Appendix 1. Voucher numbers of Charadrahyla taeniopus specimens analyzed in this study from Coleccion

Herpetologica del Centro de Investigaciones Biologicas, Universidad Autonoma del Estado de Hidalgo, México.

Year 2008, September: CIB 5422-5425 (males), December: CIB 5426 (female); 2009, April: CIB 5427 (female), June:

CIB 5428 (male), CIB 5429-5430 (females), July: CIB 5430 (female), August CIB 5431 (male), CIB 5432 (female),

October: CIB 5433-5435 (males), CIB 5436-5437 (juveniles, females), November CIB 5438 (female), CIB 5439

(juvenile, female); 2010, June: CIB 5440 (male); 2011, April CIB 5441 (female), CIB 5442-5443 (males), CIB 5444

(female), CIB 5445 (juvenile, female), CIB 5446 (male), May CIB 5447-5449 (males), June CIB 5450 (male); 2012,

March CIB 5451-5452 (females), CIB 5453 (male); 2015, April CIB 5454-5545 (males), CIB 5456 (female).

Raciel Cruz-Elizalde is a Mexican herpetologist who received his B.Sc. in Biology, M.Sc.

in Biodiversity and Conservation, and Ph.D. in Biodiversity and Conservation from the

Autonomous University of Hidalgo State, Mexico. He is currently conducting a postdoctoral

stay at the Department of Evolutionary Biology, Faculty of Science, National Autonomous

University of Mexico (UNAM). Raciel is interested in the ecology, life history evolution,

diversity, and conservation of amphibians and reptiles in Mexico. His current research includes

life history evolution of diverse lizard species of the genus Sceloporus, conservation issues in

natural protected areas, and analysis of ecological and morphological traits in the composition

of amphibian and reptile species assemblages.

Itzel Magno-Benitez is a Mexican herpetologist. Itzel recetved her B.Sc. in Biology from the

Autonomous University of Hidalgo State, Mexico, and she is interested in the ecology and

conservation of amphibians and reptiles in Mexico. Her current research interests include the

anthropic effects on conservation and ecology (feeding habits, reproduction, and morphological

variation) of amphibians and reptiles in the temperate and arid environments of Mexico.

Christian Berriozabal Islas received a Ph.D. from the Autonomous University of Hidalgo

State, Mexico. His research focuses on the diversity of amphibians and reptiles, conservation,

and the effects of climate change on tropical lizards and kinosternid turtles.

Amphib. Reptile Conserv. 20 February 2020 | Volume 14 | Number 1 | e219

Amphib. Reptile Conserv.

Cruz-Elizalde et al.

Raul Ortiz-Pulido is a Mexican scientist, amateur astronomer, scientific communicator, and

artist (landscape photographer). Ratil received a B.Sc. degree from the Universidad Veracruzana

(1994) and a doctorate in sciences from the Instituto de Ecologia, A.C. (2000). He has been

working with animal ecology since 1991, particularly the relationship between food abundance

and individuals of different species. In 1997, Raul started the Huitzil Mexican Journal of

Ornithology and in 2002 the astronomical society of his current university. In 2005, he was

elected president of CIPAMEX, a Mexican association of scientists. Raul has written more than

200 scientific papers and given more than 250 talks, many of them popularizing science. He is a

reviewer for 27 indexed scientific journals and associate editor of four of them. Currently, Raul

is associated with the Population Ecology Laboratory of the Universidad Autonoma del Estado

de Hidalgo, where he has worked since 2001.

Aurelio Ramirez-Bautista began his herpetological career conducting research as an

undergraduate student at the Los Tuxtlas Biological Field Station, Veracruz, Mexico. Aurelio

received his B.Sc. in Biology from Universidad Veracruzana in Veracruz, Mexico, his M.Sc.

and Doctorate degrees from the Universidad Nacional Autonoma de México (UNAM), and a

postdoctoral appointment at the University of Oklahoma, Norman, Oklahoma, USA. Aurelio’s

main research involves studies on ecology, demography, reproduction, conservation, and

life history evolution, using amphibians and reptiles of Mexico as models. He was president

of the Sociedad Herpetologica Mexicana and is currently associate editor of Mesoamerican

Herpetology. Aurelio is a professor at Universidad Autonoma del Estado de Hidalgo (UAEH),

teaching population ecology, herpetology, and natural history of amphibians and reptiles.

Raquel Hernandez-Austria is a biologist with an M.Sc. in Biodiversity and Conservation

Sciences from the Universidad Autonoma del Estado de Hidalgo, Mexico. Raquel’s interest is

the study of amphibian and reptile diversity and conservation. For her undergraduate degree, she

compared the food habits of two syntopic species of Lithobates, and for her Master’s degree she

evaluated the presence of Batrachochytrium dendrobatidis in anuran species of Hidalgo State.

Raquel has participated in various Mexican congresses and is an author and co-author of many

scientific papers related to amphibians and reptiles.

21 February 2020 | Volume 14 | Number 1 | e219

Official journal website:

amphibian-reptile-conservation.org

Amphibian & Reptile Conservation

14(1) [General Section]: 22—28 (e220).

“eptile-conse

Detection of Ophidiomyces ophiodiicola at two mid-Atlantic

natural areas in Anne Arundel County, Maryland and Fairfax

County, Virginia, USA

‘*Lauren D. Fuchs, ?Todd A. Tupper, *Robert Aguilar, ‘Eva B. Lorentz, ?Christine A. Bozarth,

2David J. Fernandez, and *David M. Lawlor

'George Mason University, Department of Systems Biology, 10900 University Blvd, Manassas, Virginia 20110 USA *Northern Virginia Community

College, Division of Math, Science, Technologies and Business, 5000 Dawes Avenue Alexandria, Virginia 2231 USA °*Smithsonian Environmental

Research Center, Fish and Invertebrate Ecology Lab, 647 Contees Wharf Road, Edgewater, Maryland 21037 USA *Huntley Meadows Park, Natural

Resource Management Division, 3701 Lockheed Blvd Alexandria, Virginia 22306 USA

Abstract.—Since the early 2000s, ophidiomycosis has been reported with increasing frequency and associated

with widespread morbidity in numerous North American snake species. Ophidiomyces ophiodiicola (Oo), the

etiologic agent of ophidiomycosis, has been detected in over 30 species throughout most of the eastern United

States, as well as in Europe and Australia; however, it is suspected that the distribution of this pathogen may

be underestimated due to a lack of standardized inventories. To contribute to the existing but limited data on

ophidiomycosis in the mid-Atlantic United States, snakes were sampled for Oo at two natural areas in this

region—one in Anne Arundel County, Maryland and one in Fairfax County, Virginia. Ophidiomyces ophiodiicola

was detected at both study sites. Thirty-four of 61 (55.7%) samples across eight species tested positive

for the pathogen, with the highest detection rates occurring in Nerodia sipedon (73.1%) and Pantherophis

alleghaniensis (70%). Ophidiomyces ophiodiicola was detected in snakes with (71.4%) and without (34.6%)

Clinical signs of ophidiomycosis. These results support the need for both increased Oo monitoring throughout

the region, and implementation of more standardized and unbiased sampling protocols.

Keywords. Colubridae, ophidiomycosis, population decline, Reptilia, Serpentes, snake fungal disease

Citation: Fuchs LD, Tupper TA, Aguilar R, Lorentz EJ, Bozarth CA, Fernandez DJ, Lawlor DM. 2020. Detection of Ophidiomyces ophiodiicola at two

mid-Atlantic natural areas in Anne Arundel County, Maryland and Fairfax County, Virginia, USA. Amphibian & Reptile Conservation 14(1) [General

Section]: 22—28 (e220).

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 April 2019; Accepted: 5 January 2020; Published: 12 February 2020

Introduction

Ophidiomycosis has emerged as a growing threat to

snakes throughout much of North America (Dolinski et

al. 2014; Allender et al. 2015; Lorch et al. 2016; Paré and

Sigler 2016) and has been associated with widespread

morbidity in numerous species (Guthrie et al. 2016:

Lorch et al. 2016; Stengle 2018). The disease is attributed

to Ophidiomyces ophiodiicola (Oo), a mycotic pathogen

that is only known to infect snakes (Allender et al.

2015; Lorch et al. 2016; Paré and Sigler 2016). Clinical

manifestations of infection (see Fig. 1) typically include

scabs, crusty scales, superficial pustules, subcutaneous

nodules, and dysecdysis (Dolinski et al. 2014; McBride

et al. 2015; Tetzlaff et al. 2015). Ophidiomycosis

infections are generally chronic, but mild; however,

severe infections with high mortality have been reported

in several viperid species (Allender et al. 2013; Sigler et

Correspondence. */fuchs@masonlive.gmu.edu

Amphib. Reptile Conserv.

al. 2013; Sleeman 2013; Lorch et al. 2015, 2016; Stengle

et al. 2018). The precise mechanisms that influence lethal

outcomes of the disease are still unclear, but are likely

multifaceted (Lorch et al. 2015; Guthrie et al. 2016).

Since 2006, ophidiomycosis has been increasingly

documented, with cases of infection reported in at least

20 states, including Maryland and Virginia (Allender et

al. 2015; Guthrie et al. 2016; Tupper et al. 2015, 2018,

2019). Despite growing reports of ophidiomycosis

throughout the mid-Atlantic, systematic studies

designed to assess its prevalence using non-incidental

sampling methods are limited. The rising incidence of

ophidiomycosis, coupled with habitat loss, pollution, and

other anthropogenic stressors, poses an added challenge

for snake conservation, underscoring the significance

of ongoing disease monitoring (Franklinos et al. 2017;

Kucherenko et al. 2018). The objective of this study was

to assess the presence and prevalence of Oo at two mid-

February 2020 | Volume 14 | Number 1 | e220

Fuchs et al.

Fig. 1. The ventral scales of a symptomatic Northern Black

Racer (Coluber constrictor) infected with Ophidiomyces

ophiodiicola. This 41 g male was captured and swabbed on

20 May 2018 at Huntley Meadows Park. Its total length was

123.2 cm and snout-to-vent length was 94.6 cm. Photo by Eva

Lorentz.

Atlantic natural areas located in Maryland and Virginia,

USA. The results obtained contribute toward an improved

understanding of the distribution and prevalence of

ophidiomycosis in the region.

Materials and Methods

Area-constrained visual encounter searches (Crump

and Scott 1994) were used to sample for Oo in snakes

from Huntley Meadows Park (HMP; 38°45’36.57” N,

77°05’44.13” W; Fig. 2) in Fairfax County, Virginia,

and at the Smithsonian Environmental Research Center

(SERC; 38°53717.41”N, 76°33’ 15.52” W; Fig. 3) in Anne

Arundel County, Maryland, between 22 April 2018 and

9 October 2018. Snakes were hand-captured (wearing

sterile nitrile gloves) and visually inspected for clinical

signs of ophidiomycosis (Allender et al. 2011; Clark et

al. 2011). Then, using a modified protocol developed

by Allender et al. (2016), snake skins were sampled

with sterile dry swabs (no. MW113, Medical Wire and

Equipment Company, Durham, North Carolina, USA)

from all craniofacial scales and along the entire ventral

length of the body separately, swabbing each region five

times, taking care to swab any lesions, pustules, nodules,

or displaced scales on snakes which showed signs of

infection (Allender et al. 2011, 2016). Swabs were stored

in sterile 1.5 mL microcentrifuge tubes and kept frozen

until molecular analysis. Prior to release, each snake

was measured, weighed, and photographed to help in

differentiating conspecifics. Aseptic techniques were

employed and appropriate biosecurity protocols were

followed (see Rzadkowska et al. 2016; VHS 2016) to

limit the transmission of Oo.

For the Oo assay, DNA was eluted from the swabs

using the Purification of Total DNA from Animal

Tissues Protocol (Qiagen®, Valencia, California, USA).

To ensure samples were not contaminated during the

Amphib. Reptile Conserv.

Oe 05.0% | 2

a es Kilometers

Fig. 2. Location of Huntley Meadows Park (HMP) snake

capture locations. Black markers = all samples positive; white

markers = all samples negative; gray markers = samples either

positive or negative.

extraction process, a negative control was used, which

included all elements of the extraction mixture other than

DNA. Following methods described by Allender et al.

(2015), 2.5 uL of eluted DNA was combined with 12.5

uL Sso Advanced™ universal probes supermix (Bio-

Rad, Hercules, California, USA), 1.25 uL of a combined

target-specific primer (OphioITS-F and OphiolTS-R)-

probe, and water, creating a 25 uL reaction mixture. The

DNA was amplified via GPCR using a CFX96 Touch™

Real-Time PCR Detection System (Bio-Rad, Hercules,

California, USA), with the following cycling parameters:

1 cycle at 50 °C for 2 min, 1 cycle at 95 °C for 10 min, 40

cycles of 95 °C for 15 sec and 60 °C for 60 sec, followed

by a final cycle at 72 °C for 10 min.

For each round of qPCR, a positive control was

included by adding 2.5 uL of a plasmid containing Oo

(obtained from the Wildlife Epidemiology Laboratory at

Illinois University at Urbana-Champaign, Illinois, USA)

to a designated well containing the 22.5 uL mixture of

primer-probe, and water (as described above). A well was

also included for the negative control, which contained

only the 22.5 uL mixture, but no DNA. These controls

were used to determine whether the reaction mixture

was prepared accurately, and to ensure that samples

were not contaminated during qPCR preparation. Up to

five rounds of qPCR were performed for each sample. A

sample was considered positive if at least three rounds

(per sample) had a lower cycle threshold (C,) than the

February 2020 | Volume 14 | Number 1 | e220

Ophidiomycosis in the mid-Atlantic USA

: 2

es Kilometers

Fig. 3. Location of Smithsonian Environmental Research

Center (SERC) snake capture locations. Black markers = all

samples positive; white markers = all samples negative; gray

markers = samples either positive or negative.

lowest detected standard dilution for the positive control

(Allender et al. 2016).

Sampling sites within the study areas (indicating

locations of positive and negative samples) were plotted

with ESRI ArcMap (version 10.6). Snake nomenclature

corresponds with Crother et al. (2017). Tables and

descriptive statistics were completed with Microsoft

Excel for Office 365 (Microsoft Corporation, Redmond,

Washington, USA).

Results

Sixty snakes (35 from HMP and 25 from SERC) across

nine species were captured and swabbed (Table 1).

Northern Watersnake (Nerodia sipedon) comprised the

largest proportion (n = 26; 43.3%) of the captures. Eastern

Ratsnake (Pantherophis alleghaniensis, n= 10), Common

Ribbonsnake (Thamnophis sauritus, n = 9), and Eastern

Wormsnake (Carphophis amoenus, n = 7) were also well-

represented, comprising 16.7%, 15%, and 11% of the total

snake sample, respectively. Northern Black Racer (Coluber

constrictor, n = 2), Eastern Kingsnake (Lampropeltis

getula; n = 1), Northern Ring-necked Snake (Diadophis

punctatus, n = 1), Eastern Gartersnake (Thamnophis

sirtalis, n= 2), and Dekay’s Brownsnake (Storeria dekayi;

n = 2) were all sparsely represented. Ophidiomyces

ophiodiicola was detected in 33 snakes and in a shed skin

of a Northern Black Racer, yielding an overall detection

rate of 55.7%. More than half of the positive samples

(55.9%) were from a single species—Northern Watersnake.

Of the nine species sampled, Northern Watersnake had

the highest detection rate (73.1%), followed closely by

Eastern Ratsnake (70%). Northern Black Racer, Eastern

Wormsnake, and Common Ribbonsnake were positive in

66.7%, 28.6%, and 11.1% of samples, respectively. Only

one Eastern Kingsnake and one Northern Ring-necked

snake were sampled, and both were positive. Dekay’s

Brownsnake was positive in one of two samples and

Eastern Gartersnake was the only species that did not test

positive for Oo. Twenty-five of the 35 (71.4%) snakes

showing clinical signs tested positive for Oo, and nine of

the 26 (34.6%) without clinical signs were Oo positive

(Table 1). Prevalence varied between study locations,

with 34.6% of snakes testing positive at HMP and 84.6%

at SERC. Of the 34 snakes testing positive, Oo was

detected in both swabs in 18 snakes (52.9%) and in only

one of two swabs (nine from the craniofacial swab only,

seven from the body swab only) in 16 snakes (47.1%).

Table 1. Prevalence by species. S/+ = positive with clinical signs, A/+ = positive without clinical signs.

Species

Eastern Wormsnake (Carphophis amoenus amoenus)

Northern Black Racer (Coluber constrictor constrictor)

Northern Ring-Necked Snake (Diadophis punctatus

edwardsii)

Eastern Kingsnake (Lampropeltis getula)

Northern Watersnake (Nerodia sipedon sipedon)

Eastern Ratsnake (Pantherophis alleghaniensis)

Dekay's Brownsnake (Storeria dekayi)

Common Ribbonsnake (Thamnophis saurita saurita)

Eastern Gartersnake (Thamnophis sirtalis sirtalis)

Total or overall prevalence

Amphib. Reptile Conserv.

Prevalence (%)

N_ Positive Forspecies Overall S/+ A/+

7 2 28.6 59 0 es

3 2 66.7 5.9 2 0

1 1 100 2.9 0 1

1 1 100 2.9 1 0

26 19 73.1 559 15 4

10 7 70 20.6 > 2

2 1 50 2.9 1 0

9 1 11.1 2.9 1 0

2 0 0 0 0 0

61 34 - 55.7 25 9

February 2020 | Volume 14 | Number 1 | e220

Fuchs et al.

Discussion

Although Oo has previously been documented in

Maryland and Virginia (Guthrie et al. 2016; Tupper et al.

2018), this work is one of only two studies (see Guthrie

et al. 2016) to investigate Oo in these states. In Maryland,

observations of fungal dermatitis have been reported

from the Smithsonian Environmental Research Center

(SERC) since 2014 (Tupper et al. 2015), with Oo recently

being confirmed as the etiological agent of a dermal

infection in Northern Watersnake (Tupper et al. 2018).

These results add four new species (Eastern Wormsnake,

Northern Black Racer, Northern Ring-necked Snake, and

Eastern Ratsnake) to the documented host range of this

pathogen in Maryland, which previously included only

Northern Watersnake (Tupper et al. 2018) and Timber

Rattlesnake (Crotalus horridus;, Tupper et al. 2019). In

eastern Virginia, Guthrie et al. (2016) documented Oo in

four species (all with clinical signs): Northern Watersnake

(n= 3), Rainbow Snake (Farancia erytrogramma; n= 1),

Northern Black Racer (n = 2), and Brown Watersnake

(Nerodia taxispilota, n = 2). This study adds four new

hosts to the list of Oo positive species occurring in

Virginia: Eastern Kingsnake, Eastern Ratsnake, Dekay’s

Brownsnake, and Common Ribbonsnake.

The overall detection rate of 57.4% is among the

highest reported (except see McKenzie et al. 2018)

across the eastern and midwestern United States (Smeenk

et al. 2016; Allender et al. 2016). The prevalence of

Oo throughout these regions appears to be highly

variable, with detection rates as low as 0% and 4.9% in

Ohio and Michigan, respectively (Smeenk et al. 2016;

Allender et al. 2016), and up to nearly 62% in eastern

Kentucky (McKenzie et al. 2018). We interpret these

rates cautiously, however, taking into consideration the

variation in species sampled between studies. It is still

unclear how susceptibility and severity of infection differ

between species (Grisnik et al. 2018), but the composition

of species sampled in this study may partly explain the

overall prevalence and the relatively high proportion of

Oo positive snakes that did not show clinical signs of the

disease.

In this study, Oo was detected in eight of the nine

species sampled, which was not surprising given that

each of these species has previously tested positive for

the pathogen in the eastern and mid-western United

States (Lorch et al. 2016; Persons et al. 2017; Grisnik

et al. 2018; McKenzie et al. 2018). However, the small

sample sizes in certain species made it impossible to

assess how each of these species actually influence the

overall detection rate. Ophidiomyces ophiodiicola was

found to be most prevalent in Northern Watersnake, with

a detection rate of 73%. This species represented nearly

43% of the total sample and thus had a strong influence

on overall prevalence (55.7%). Prior studies with similar

proportions of aquatic species have also demonstrated

relatively high Oo detection rates among Northern

Amphib. Reptile Conserv.

Watersnakes and other species with aquatic affiliations.

However, this trend in detection may partially reflect the

habitat preferences of the pathogen (Lorch et al. 2016;

McKenzie et al. 2018), rather than an inherent biological

susceptibility to the pathogen. Additional work is needed

to better understand susceptibility to the disease.

Variability in sampling methods between studies

should also be considered when interpreting results

(McCoy et al. 2017; Grisnik et al. 2018; Hileman et al.

2018; McKenzie et al. 2018). For instance, the number

of sterile dry swab applicators used per snake has been

shown to influence detectability of Oo, with the use of

only one applicator greatly increasing the probability

of obtaining false-negatives (Hileman et al. 2018). The

results obtained here support this concept, with 47.1% of

snakes testing positive for Oo in only one of two swabs.

Underestimation of the prevalence of Oo may also

occur when diagnostic tests are limited only to snakes that

present clinical manifestation of infection (see Guthrie et

al. 2016). While clinical signs have been associated with

a higher probability of PCR-positive results (Allender et

al. 2016), studies have also demonstrated that anywhere

from 6% (Bohuski et al. 2015) to 38% (Hileman et al.

2018) of snakes without clinical signs test positive for Oo.

The data reported here support these studies, with 26.5%

of Oo positive snakes in this sample showing no signs of

infection. One possible explanation is that clinical signs

may be subtle and overlooked during inspection, because a

snake is either in the early stages of infection or effectively

clearing the infection through repeated sheds (Lorch et al.

2016; Grisnik et al. 2018; Hileman et al. 2018). Detection

without clinical signs may also reflect the absence

of infection in a specimen altogether. Ophidiomyces

ophiodiicola can persist as a saprobe in the soil, which

can facilitate transmission and increase the likelihood of a

snake encountering Oo (Allender et al. 2015; Lorch et al.

2016). The presence of Oo on the skin, however, does not

necessarily indicate infection. Therefore, while swabbing

can be an effective, low-cost, and minimally invasive

method for detecting the pathogen, it cannot be used to

infer or imply infection status.

Results from this study confirm that Oo is present and

relatively prevalent in both Maryland and Virginia, and

that the presence of Oo is more often accompanied by

clinical manifestations consistent with ophidiomycosis

than not. The geographic distribution and host range

of the pathogen are still largely unknown (Burbrink

et al. 2017), and ophidiomycosis may be more widely

distributed than documented cases suggest (USGS 2018).

Some have proposed that biased sampling methods

may result in underestimations of prevalence within a

population (Grisnik et al. 2018; Hileman et al. 2018). This

potential for inaccurate assessments highlights the need

for more standardized sampling efforts and diagnostic

protocols. Based on the increasing number of reports of

ophidiomycosis throughout the eastern United States,

we suggest increased efforts to identify and monitor

February 2020 | Volume 14 | Number 1 | e220

Ophidiomycosis in the mid-Atlantic USA

Oo throughout the mid-Atlantic region. Additionally,

enhanced biosecurity protocols should be implemented

to limit disease transmission throughout the region.

Acknowledgements.—This work was funded by a

NOVA Foundation Innovation Grant. Valerie Czach,

Amanda Lee, and Ashely Davis assisted with field and

lab work. Maggie Emblom-Callahan, Susan Williams,

Tatiana Stantcheva, and the NOVA Police Department

provided logistical support. This work was approved

by the Institutional Animal Care and Use Committee at

George Mason University (reference #0396), the Virginia

Department of Game and Inland Fisheries (permit #

062364), and the Maryland Department of Natural

Resources (permit #57025).

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Amphib. Reptile Conserv.

Lauren Fuchs is a Ph.D. student in Environmental Science and Policy at George Mason

University (Fairfax, Virginia, USA). Lauren is also a graduate teaching assistant, and volunteer

at the Smithsonian Environmental Research Center (Edgewater, Maryland, USA). Her main

area of interest is in the diseases of herpetofauna.

Todd Tupper is a Professor of Biology at Northern Virginia Community College (Alexandria,

Virginia, USA), an affiliate faculty member at George Mason University (Fairfax, Virginia,

USA), and a Visiting Researcher at the Smithsonian Environmental Research Center (Edgewater,

Maryland, USA). Todd teaches general biology, zoology, and biostatistics. His areas of interest

are biology education, and amphibian and reptile monitoring and conservation.

Robert Aguilar is a biologist at the Smithsonian Environmental Research Center (Edgewater,

Maryland, USA). His areas of interest are quite varied and include fish and invertebrate ecology

and phylogenetics, herpetological diseases, wildlife inventory and monitoring, and science

education. Robert helps with the Maryland Biodiversity Project and supervises undergraduate

researchers at the Smithsonian Environmental Research Center.

27 February 2020 | Volume 14 | Number 1 | e220

Amphib. Reptile Conserv.

Ophidiomycosis in the mid-Atlantic USA

Eva Lorentz is an undergraduate senior in Biology at George Mason University (Fairfax, Virginia,

USA) and a volunteer for the Smithsonian Environmental Research Center (Edgewater, Maryland,

USA).

Christine Bozarth is an Associate Professor of Environmental Science at Northern Virginia

Community College (Alexandria, Virginia, USA). Her areas of interest are in population genetics,

wildlife inventory and monitoring, and science education.

David Fernandez is a Professor of Biology at Northern Virginia Community College (Alexandria,

Virginia, USA). David teaches cell biology, general biology, and anatomy and physiology. His

areas of interest are in hepatic cancers and wildlife disease. His areas of interest are in hepatocyte

biology, tumor biology, and herpetological diseases.

David Lawlor is a biologist at Huntley Meadows Park in Alexandria, Virginia, USA. His areas

of interest are in natural history, natural resource management, and science education. David also

coordinates and oversees the wildlife research that occurs at Huntley Meadows.

28 February 2020 | Volume 14 | Number 1 | e220

Official journal website:

amphibian-reptile-conservation.org

Amphibian & Reptile Conservation

14(1) [General Section]: 29-42 (e221).

AEs

le-conse*

Insights into the natural history of the endemic Harlequin

Toad, Atelopus laetissimus Ruiz-Carranza, Ardila-Robayo,

and Hernandez-Camacho, 1994 (Anura: Bufonidae), in the

Sierra Nevada de Santa Marta, Colombia

'*Hernan D. Granda-Rodriguez, 7Andrés Camilo Montes-Correa, *Juan David Jiménez-Bolafo,

‘Alberto J. Alaniz, Pedro E. Cattan, and °Patricio Hernaez

'Programa de Ingenieria Ambiental, Facultad de Ciencias Agropecuarias, Universidad de Cundinamarca, Facatativa, COLOMBIA *%Grupo de

Investigacion en Manejo y Conservacion de Fauna, Flora y Ecosistemas Estratégicos Neotropicales (MIKU), Universidad del Magdalena, Santa Marta,

COLOMBIA ‘Centro de Estudios en Ecologia Espacial y Medio Ambiente, Ecogeografia, Santiago, CHILE °Facultad de Ciencias Veterinarias y Pecuarias,

Universidad de Chile, Santiago, CHILE °Centro de Estudios Marinos y Limnologicos, Facultad de Ciencias, Universidad de Tarapaca, Arica, CHILE

Abstract.—Atelopus laetissimus is a bufonid toad that inhabits the mountainous areas of the Sierra Nevada de

Santa Marta (SNSM), Colombia. This species is endemic and endangered, so information about its ecology and

distribution are crucial for the conservation of this toad. Here, the relative abundance, habitat and microhabitat

uses, and vocalization of A. /laetissimus are described from the San Lorenzo creek in the SNSM, as well as its

potential distribution in the SNSM. To this end, 447 individuals were analyzed during several sampling trips

from 2010 to 2012. Against expectations, population density was significantly higher in the stream than in the

riparian forest. Overall, A. laetissimus used seven different diurnal microhabitats, with a high preference for

leaf litter substrates and rocks. The rate of recaptures decreased linearly across the survey nights. Two types

of vocalizations related to the advertisement call of A. latissimus were recorded: a series of pulsed calls like

a buzz and another short call, lacking pulses or partially pulsed. According to this analysis, the areas with

higher habitat suitability for A. laetissimus were located principally in the northern and northwestern regions

of the SNSM, in agreement with literature. Moreover, the data modeling indicated a significant increase in

habitat loss from 2013 to 2017. The information presented here should be considered as a starting point for the

conservation of this species.

Keywords. Advertisement call, amphibian decline, conservation, ecology, habitat loss, habitat suitability, home range,

microhabitat selection, nocturnal site fidelity

Resumen.—Atelopus laetissimus es un sapo de la familia Bufonidae que habita las zonas montanosas de la

Sierra Nevada de Santa Marta (SNSM), Colombia. Para esta especie endemica y en peligro, la informacion

ecologica y de distribucion es crucial para su conservacion. En el presente trabajo describimos la abundancia

relativa, usos de habitat y microhabitat, y las vocalizaciones de A. laetissimus, asi como su distribucion

potencial en la SNSM. Para esto, analizamos 477 individuos durante varios muestreos entre 2010 y 2012.

Contra las expectativas, la densidad poblacional fue significativamente mayor en el lecho de la quebrada que

en el bosque ribereno adyacente. En general, A. laetissimus utilizo siete microhabitats diurnos, con una alta

preferencia por los sustratos de hojarasca y rocosos. La tasa de capturas decrecio linealmente a lo largo de

los muestreos nocturnos. Registramos dos tipos de vocalizaciones relacionadas con el llamado de anuncio

de A. laetissimus. Una serie de Ilamados pulsados como zumbidos y otros mas cortos, con pulsos ausentes

oO parcialmente pulsados. De acuerdo con nuestros analisis, las areas con mayor idoneidad de habitat se

localizan en los sectores septentrionales y noroccidentales de la SNSM, lo que es concordante con la literatura.

Ademas, el modelo construido indica un incremento significativo de la pérdida de habitat entre 2013-2017.

Esta informacion debe ser considerada como punto de partida para la conservacion de esta especie.

Palabras clave. Ambito doméstico (home range), conservacion, declive de los anfibios, ecologia, fidelidad de percha

nocturna, idoneidad de habitat, llamado de anuncio, pérdida de habitat, seleccion de microhabitat

Citation: Granda-Rodriguez HD, Montes-Correa AC, Jiménez-Bolafio JD, Alaniz AJ, Cattan PE, Hernaez P. 2020. Insights into the natural history of

the endemic Harlequin Toad, Atelopus /aetissimus Ruiz-Carranza, Ardila-Robayo, and Hernandez-Camacho, 1994 (Anura: Bufonidae), in the Sierra

Nevada de Santa Marta, Colombia. Amphibian & Reptile Conservation 14(1) [General Section]: 29-42 (e221).

[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: 3 April 2019; Accepted: 31 October 2019; Published: 17 February 2020

Correspondence. !*hernangrandar@gmail.com;, *andresc.montes@gmail.com, * herpetos4@gmail.com; * alberto.alaniz@ug.uchile.cl,

° pahernaez@gmail.com

Amphib. Reptile Conserv. 29 February 2020 | Volume 14 | Number 1 | e221

Natural history of Harlequin Toad, Atelopus /aetissimus in Colombia

Introduction

The harlequin toads (Bufonidae: Ate/opus) are small

amphibians (<10 cm) which have aposematic coloration

and predominantly diurnal activity periods (Lotters

1996). With a worldwide diversity of 96 described species

(Frost 2019), the conservation of these amphibians has

been seriously affected during the last decades due to the

population decreases detected in a majority of species

and the extinction of others, and currently 97.92% of

the species of this genus are included in the IUCN Red

List of Threatened Species, with one extinct species

(Young et al. 2001; La Marca et al. 2005; Gascon et al.

2007; Tapia et al. 2017; IUCN 2019). Among the main

risk factors for the decline of these populations are

the loss of habitat, the introduction of exotic species

which are potential predators and competitors, as well

as deaths by pathogens (mainly by the chytrid fungus

Batrachochytrium dendrobatidis [Bd]|) and _ climate

change (Lotters 2007; Catenazzi 2015; Barrio-Amoros

and Abarca 2016; Valenzuela-Sanchez et al. 2017).

In general, information about the population biology

and ecology of the different Ate/opus species 1s relatively

scarce. For example, the males of the species Ate/opus

cruciger (Lichtenstein and Martens, 1856) are known to

remain longer in the streams than the females (Sexton

1958). But, on the other hand, studies conducted on

Atelopus carbonerensis (Rivero, 1974 “1972”) in

Venezuela, found that they remain almost all year in

their habitat, except in the dry season when individuals

migrate towards the streams for reproduction (Dole and

Durant 1974). In the Variable Harlequin Toad of Costa

Rica, Atelopus varius (Lichtenstein and Martens, 1856),

both males and females are territorial and have fidelity

for their reproduction sites. This indicates that even

temporary alterations of the aggregation patterns of

individuals between dry and rainy seasons, when they are

more dispersed due to the increase in humidity (Crump

1988; Pounds and Crump 1989), results in an apparent

decrease in detection (Gonzalez-Maya et al. 2013).

In Panama, the Golden Toad, Ate/opus zeteki (Dunn,

1933) 1s concentrated in streams at the beginning of the

breeding season, which occurs only during the transition

between the rainy season and the dry season (Karraker et

al. 2006). Another study determined site fidelity, habitat

utilization, and range of households in the Suriname

Toad, Atelopus hoogmoedi (Lescure, 1974) during the

rainy season (Luger et al. 2009).

The Sierra Nevada of Santa Marta (SNSM) is

a mountain massif located in the Caribbean region

of Colombia. The particular conditions of isolation

and vegetation of this mountainous system, which

is not connected to the Andes, have led to a series of

speciation processes in several groups of vertebrates,

such as amphibians and reptiles (Ruthven 1922; Bernal-

Carlo 1991; Sanchez-Pacheco et al. 2017), mammals

(Alberico et al. 2000), and birds (Strewe and Navarro

Amphib. Reptile Conserv.

2004). This applies to the harlequin toads (Bufonidae:

Atelopus Dumeéril and Bibron, 1841), whose current

diversity includes five endemic species for this region:

Atelopus arsyecue Rueda-Almonacid, 1994, Atelopus

carrikeri Ruthven, 1916, Atelopus laetissimus Ruiz-

Carranza, Ardila-Robayo, and Hernandez-Camacho,

1994, Atelopus nahumae Ruiz-Carranza, Ardila-Robayo,

and Hernandez-Camacho, 1994, and Atelopus walkeri

Rivero, 1963. These species can be found between 800

and 4,500 m asl, and from the tropical moist forests to the

paramos of the SNSM (Ruthven 1916; Rueda-Almonacid

1994; Ruiz-Carranza et al. 1994; Rueda-Almonacid et al.

2005).

This study examines some aspects of the natural

history and ecology of the Harlequin Toad, A. /aetissimus.

Previous studies have shown that A. /aetissimus inhabits

streams and rivers in the mountainous areas of the

northwestern sector of the SNSM (Granda-Rodriguez

et al. 2008, 2012; Rueda-Solano et al. 2016a), and it is

classified as Endangered (EN) by the IUCN (Granda et

al. 2008). A recent study reported individuals infected by

the chytrid fungus Bd (Flechas et al. 2017), the pathogen

that has led to the decline and disappearance of many

amphibian populations globally (Young et al. 2001;

Catenazzi 2015). This study also includes an estimation

of relative abundance, population density, microhabitat

preference, spatial dynamics, and vocalization, in a

population of A. /aetissimus in the sector of San Lorenzo,

Santa Marta, Colombia. Additionally, the potential

distribution and patterns of habitat loss for this species

during the 21" century were modeled, recognizing the

great influence of habitat loss in recent decades on the

extinction of many Neotropical species (Young et al.

2001; Marca et al. 2005; Lotters 2007).

Materials and Methods

Study Area

The study area corresponds to the San Lorenzo creek

(11°6’56.21” N, 74°3’0.18” W, 2,100 m asl), an affluent

of the upper basin of Gaira river, northwestern sector of

the SNSM, Santa Marta district, Magdalena department

(= state), Colombian Caribbean. The principal vegetation

unit of this area corresponds to lower mountain humid

forest (sensu Espinal and Montenegro 1963). According

to Granda-Rodriguez et al. (2012), the annual averages

of rainfall and temperature are 2,622 mm and 13.6 °C,

respectively, and the climatic regime is unimodal bi-

seasonal, with a dry period from December to March,

and a rainy period from April to November.

Estimations of Relative Abundance and Population

Density

Relative abundance (RA) was estimated from data col-

lected in seven field campaigns conducted from 2010 to

February 2020 | Volume 14 | Number 1 | e221

Granda-Rodriguez et al.

2012 (October 2010; April, June, and December 2011;

January and February 2012). Each fieldtrip had a duration

of 12 days, with seven hours of daily work (0800-1200 h,

1800-2100 h). Individuals were detected through Visual

Encounter Survey (Crump and Scott 1994), where two

observers performed random walks. The sampling effort

was 84 h per observer for each field campaign, reach-

ing 558 h per observer in total. Relative abundance was

calculated as the number of individuals/(h < observers),

or ind/[h x obs] (Lips 1999). Sex was assigned by the

size of individuals, assuming that females had a snout-

vent length (SVL) > 40 mm and juveniles < 35 mm, and

specimens within this range were considered as potential

males. Sexual determination also considered the pres-

ence of eggs in the corporeal cavity noted through skin,

calling behavior, and amplectant couples. Sex was deter-

mined in this way because the sexually dimorphic char-

acters typically useful for population studies (La Marca

et al. 1990 “1989;” Lampo et al. 2017) have not been

established for A. /aetissimus.

Population density was estimated through 40 perma-

nent transects of 20 x 4 m (Jaeger 1994), with 20 located

in the riparian forest and 20 in the stream. These tran-

sects were positioned parallel to the stream, separated

by at least 20 m. Two observers walked along the tran-

sect counting the individuals only once. The medians of

density obtained in each riparian forest and stream were

compared using the Wilcoxon test for independent sam-

ples (W).

Habitat Selection and Dispersal Patterns

Testing the microhabitat preferences followed the con-

cept of the third and fourth levels of habitat selection

according to Johnson (1980), that indicate which com-

ponents of the habitat are used and their proportions of

use. For diurnal microhabitat, the substrate occupied by

each individual was recorded according to the seven cat-

egories proposed by Granda-Rodriguez et al. (2008b): (1)

rocks, (II) leaf litter, (III) fallen trunks, (IV) ferns, (V)

leaves, (VI) bare floor, and (VII) others. The proportion

of area for each microhabitat category was measured in

15 random plots of 4 m? located at the side of the stream.

A Chi-squared test (y) was performed, where the expect-

ed frequency was the number of individuals by substrate

(N), while the observed frequency was calculated as the

total number of individual (N) per area proportion of the

substrate (%). Microhabitat selection was assumed when

the proportion of substrate used was different from its

availability, following the method of Molina-Zuluaga

and Gutiérrez-Cardenas (2007). To determine which

substrates were selected by individuals, this analysis was

repeated after deleting the categories most used or those

that seemed to be used disproportionally to their avail-

ability (Molina-Zuluaga and Gutiérrez-Cardenas 2007).

In cases where significant differences in the second anal-

Amphib. Reptile Conserv.

ysis were not found, the deleted category was considered

to be preferred by the species.

To determine the nocturnal site fidelity, 60 speci-

mens were marked with sub-epidermal alphanumeric

tags (Visible Implant Alpha Tags, Northwestern Marine

Technology Inc., 1.5 x 2.5. mm), and detected at night

with a fluorescence lantern (Courtois et al. 2013). Dur-

ing 13 continuous nights (1900-2200 h, 36 h x obs), two

observers looked for marked individuals to determine if

they stayed in the same sites. The date, hour, location,

and distance from the previous capture site were record-

ed for each recapture. The potential relationship between

the number of individuals recaptured and the number of

nights of survey was explored using a linear regression.

To describe the patterns of diurnal horizontal move-

ment, each marked and recaptured specimen was spa-

tially located in a Cartesian diagram consisting of a 50 m

transect along the stream delimited by a reference point

every 5 m (y axis), and the perpendicular distance of the

specimen to the transect (x axis). Then, the distance from

diurnal to nocturnal microhabitat was measured when

possible. This was carried out in two sampling sessions of

three days, with a three-day interval between them. The

sampling times on the first day were 0800-1100 h and

1400-1600 h, on the second at 0600-0900 h and 1300-—

1500 h, and on the third at 0900-1200 h and 1500-1700

h. This sequence was repeated successively. Aggressive

behavior observed during the survey was described fol-

lowing the terminology of Crump (1988).

Advertisement Call

On 30 January 2012, 270 seconds (s) of the advertisement

call (sensu Wells 2007) of a male of A. /aetissimus was

recorded in San Lorenzo creek, at 1732 h, using a Sony

(ICD-PX312) digital recorder. Although the advertisement

call is not easy to define, we consider that the recorded calls

belong to this functional category because they were emit-

ted regularly by a solitary male in situ (without manipula-

tion), who had no interactions with individuals of the same

sex (which might indicate aggressive calls) or the opposite

sex (which might indicate courtship calls). Air tempera-

ture and relative humidity at the recording moment were

12.3 °C and 75%, respectively. The traits of advertisement

calls were quantified using the software PRAAT 6.0.13 for

Windows (Broesma and Weenik 2007). The parameters

of the advertisement call measured were: call duration (in

seconds: s), number of pulses per call, pulse duration (s),

interpulse interval (s), rate of pulses per second (pulses/s),

frequency range (Hz), dominant frequency (Hz), and vis-

ible harmonics (Hz). Means and standard deviations (SD)

were calculated for each call parameter. The terminology

proposed and revised by Cocroft et al. (1990) was used for

call types and by Kohler et al. (2017) for call parameters.

Spectrograms and oscillograms were generated using the

Seewave package (Sueur et al. 2008) in R environment (R

Core Team 2018).

February 2020 | Volume 14 | Number 1 | e221

Natural history of Harlequin Toad, Atelopus /aetissimus in Colombia

Relative abundance (ind/[h x obs])

0.0-

2010-VIl 2014-IIl 2041-VI 2011-XI 2011-XII 2012-1 2012-II

Oy Pe

0.00-

Forest Stream

Fig. 1. (A) Temporal variation of relative abundance (ind/[h x obs]). (B) Population density (m7) per habitat of Atelopus laetissimus.

Roman numerals represent the months of the surveys.

Potential Distribution and Habitat Loss

A species distribution model was performed, which pre-

dicts the habitat suitability with predictive algorithms

integrated from environmental data and museum re-

cords (Phillips et al. 2017). The available records of A.

laetissimus were compiled from Global Biodiversity

Information Facility (GBIF), and the authors’ own data,

considering all known records from 1969 to 2017. Nine-

teen bioclimatic layers from Worldclim 2.0. (Fick and

Hijymans 2017) plus altitude, human footprint (Venter

et al. 2016), and solar radiation were used as predictor

variables. First, data were explored through a prelimi-

nary model including all variables, with the technique

of maximum entropy (MaxEnt software, 3.4.1, Phillips

et al. 2017), considering that this algorithm is not hin-

dered by a minimum number of occurrences. Variables

with correlation indexes > 0.7 and lower contributions to

the exploratory model were removed. To reduce the over-

fitting of the model, collinearity was determined with a

Spearman correlation test. According to van Proosdjj et

al. (2016), the size of the background was considered in

relation to the prevalence of the species to be modeled

(< 25 localities), since this criterion generates acceptable

results for species with restricted distributions.

A model of seven-fold bootstrap technique was per-

formed, using 65% of data for training and 35% of data

for testing (Puschendorf et al. 2008), considering the

small number of locations that could be used (Elith et

al. 2011). The average and standard deviation of the pre-

dicted suitability were used as a final model, and as a

spatially explicit measure of the reliability of the predic-

tion, respectively. The accuracy of the model was esti-

mated using the metric of the area under the curve of the

receiver operating characteristic (AUC, Elith et al. 2011).

Amphib. Reptile Conserv.

Additionally, the distribution extent was calculated us-

ing the IUCN methodology (2019), based on the area of

occupation (AOO) and extent of occurrence (EOO). The

AOO was calculated as the intersection of the species oc-

currence with a square grid of 2 x 2 km, while EOO cor-

responded to the minimum convex polygon drawn on the

peripheral localities of the distribution area. Both AOO

and EOO were calculated using only records after 2010.

To determine habitat loss, the resulting map from the dis-

tribution model was overlapped with forest cover loss maps

from 2000 to 2017. These forest cover loss maps were gen-

erated by Hansen et al. (2013), who monitored the changes

of forest cover annually with a spatial resolution of 30 m.

This product has shown important benefits in terms of its

feasibility for evaluating the loss and fragmentation of habi-

tat for forest specialist species (Alaniz et al. 2018; Carva-

jal et al. 2018). The cumulative and annual habitat losses

were calculated for the potential distribution, AOO, and

EOO. For the annual habitat loss, fourth order polynomial

regressions were performed to test the trends of the multian-

nual habitat loss. The AOO and EOO estimations, and their

respective trends, allowed a suggested threat classification

based on criterion B of IUCN (2019) Red List of Species.

Results

Relative Abundance and Population Density

A total of 447 individuals of A. /aetissimus were record-

ed, distributed potentially among 428 males, 16 females,

and three juveniles. The numbers of individuals per

survey fluctuated from 32 to 90 (median + interquartile

range, 78 + 44.5 individuals), with a general RA of 0.38

ind/(h x obs), and 0.21 to 0.54 ind/(h x obs) for each sur-

vey (Fig. la). Atelopus laetissimus showed a population

32 February 2020 | Volume 14 | Number 1 | e221

Granda-Rodriguez et al.

30-

Recaptures (n)

5 10

Nights of survey

Transect along the stream (m)

e 5 tae

30- 4 Individuals

#| A01

S| A3B6

4 sily a| Aa

0 —

20; [a | aaa

[a | ast

3 2 lel

0- 479

0.4 0.8 1.2 1.6

Distance to stream (m)

Fig. 2. Temporal variation of the number of recaptures (A) and movement patterns (B) of Ate/opus laetissimus.

density from 0 to 0.13 ind/m? (0.04 + 0.06 ind/m7). Popu-

lation density was significantly higher (Wilcoxon test, W

= 277.5, p = 0.036, Fig. 1b) in the stream (O—0.12 ind/

m’, 0.05 + 0.07 ind/m7) than in the riparian forest (O-0.11

ind/m?, 0.03 + 0.05 ind/m?). The corresponding author

will provide tables of raw data for individual specimens

on request.

Habitat Selection and Dispersal Patterns

Atelopus laetissimus used seven different diurnal mi-

crohabitats, which were also used differentially regard-

ing their availability (y?7 = 120.121, df = 6, p < 0.001).

Although the leaf litter and rocks were the most used

substrates (Table 1), significant differences in the use of

microhabitat were still evident when these were removed

from the analysis (7 = 471.991, df = 4, p < 0.001), sug-

gesting that there is no preference for these substrates.

From 60 marked specimens, three (5%) to 31 (52%)

were recaptured per night of survey. The recapture rate

was higher than 30% until the fifth night, while after the

ninth night it was reduced to less than 7%, showing a lin-

ear decrease across the sampling nights (7° = 0.86, F_,,

= 69.99, p < 0.001, Fig. 2a). The height of the noctur-

nal microhabitat ranged between 10 and 168 cm (mean

+ SD, 73.03 + 48.41 cm). The nocturnal site fidelity of

A. laetissimus did not appear to be related to its height

or to the SVL of specimens. Only six of the 60 tagged

individuals were recaptured more than four times. These

specimens showed an average home range of 0.35 +

0.21 m? (0.1-0.59 m’, Table 2, Fig. 2b), with an aver-

age horizontal displacement of 1.92 + 0.82 m (0.8-3 m)

relative to the nocturnal site. Most of the specimens were

separated from each other by at least 5 m, but specimens

Al and A4 were very close to each other, so aggressive

behavior between them could be observed. Specimen

Amphib. Reptile Conserv.

A4 pounced and squashed specimen A1, then they be-

gan actively “wrestling.” This situation lasted about 120

seconds, until Al fled. The “winner” male (A4) did not

chase the male who left. The males did not emit vocaliza-

tions during the event.

Advertisement Call

Two types of vocalizations were recorded in a male A.

laetissimus. The first call type corresponded to a short

series of pulses like a buzz (mean + SD, range, N; 27 +

5.63 pulses, 7-33 pulses, 26 calls, Fig. 3a), with a dura-

tion of 0.41 + 0.112 s (0.111—0.805 s, 26 calls). These

pulsed calls showed modulated amplitude, where the

amplitude increased along the call and decreased again

at the last pulse. The pulse duration was 0.009 + 0.006

s (0.001— 0.099 s, 705 pulses), emitted at a rate of 67.55

+ 9.428 pulses/s (32.298—-76.167 pulses/s, 26 calls). In

most of these calls, the last pulse had a longer duration.

The interpulse interval duration was 0.006 + 0.004 s

(0.0001—0.097 s, 652 interpulse intervals). These calls

showed an ascending modulated frequency, although in

some cases the frequency decreased notably at the last

pulse. The frequency range was 1,287—8,558 Hz, while

the dominant frequency was 1,921.433 + 114.391 Hz

(1,480.95—2,155.55 Hz, 631 pulses). In addition, the

pulsed call of A. /aetissimus showed three harmonics:

first at 2,640.54-4,923 Hz, second at 3,661.56—6,644 Hz,

and third at 5,771.95-8,558 Hz. The second type was

a short call (Fig. 3b), with a duration of 0.06 + 0.006

s (0.05—0.07, 12 calls). The short calls showed a vari-

able structure, either unpulsed (Fig. 3b), partially pulsed

(Fig. 3c), or pulsed (Fig. 3d). The short calls showed a

low dominant frequency (1,649.59 + 32.715, 1,584.2-

1,705.99, 12 calls) and were produced irregularly, from

a variable series, and alternated among vocalizations of

February 2020 | Volume 14 | Number 1 | e221

Natural history of Harlequin Toad, Atelopus /aetissimus in Colombia

Table 1. Microhabitat selection by Ate/opus laetissimus and y? values for each substrate.

Substrate Proportion of area Individuals () ee Ia WRG REA x?

Rocks 0.59 49 66.22 4.48

Leaf litter 0.28 28 31319 0.33

Others 0.03 2 355 0.59

Ferns 0.01 3 0.75 6.81

Leaves 0.05 9 5.65 1.99

Bare floor 0.01 12 13 104.56

Fallen trunks 0.04 ih 4.52 1.36

N 1 113 h3 120.12

the first type (Fig. 3e), or within vocalizations of the first

type (Fig. 3f).

Potential Distribution and Habitat Loss

The areas with higher habitat suitability for A. /aetissi-

mus are located mainly in the northern and northwestern

sectors of the SNSM, which agrees with the occurrence

localities. The model reached an AUC of 0.971 (+ 0.011,

Fig. 4). The explanatory variables with the highest con-

tributions to the suitability prediction were the average

temperature of the coldest trimester (Bio 11), range of an-

nual temperature (Bio 7), elevation, and human footprint.

Atelopus laetissimus shows a maximum of suitability at

120 mm of rainfall at the coldest trimester; for annual

temperature, it shows high suitability at middle ranges,

rapidly decreasing at under 12 °C. The suitability shows

a Gaussian trend regarding elevation, with a maximum at

2,000-—3,000 m asl. Habitat suitability of A. /aetissimus is

inversely related with human footprint (Fig. 4).

The potential distribution area is 1,740.95 km’, which

corresponds to a continuous area in the northwestern sec-

tor of the SNSM, and a smaller and fragmented area in the

northern sector. The AOO calculated by 13 plots was 54

km’, while the EOO was 1,074.47 km?. The habitat loss

from 2000 to 2017 was larger in the southern sector of

the SNSM, but was smaller in the northern and northwest-

ern sectors of the massif. Ate/opus laetissimus lost 1.48%

of its habitat based on the potential distribution, 1.16%

of its AOO, and 2.51% of its EOO (Fig. 5). A significant

increasing trend in habitat loss was detected from 2013

to 2017, where the last year showed the greatest loss of

potential habitat for the species. As A. /aetissimus shows

an EOO smaller than 5,000 km? (criteria B1b[i], B1 b[ii],

and B1bfiti]) and an AOO smaller than 500 km? (criteria

B2b[i], B2b[11], and B2b/i11]), these data reinforce its clas-

sification in the Endangered (EN) category.

Discussion

Relative Abundance and Density

The results of these surveys showed that Ate/opus laetis-

simus 1S an easily detectable species in the northwestern

Frequency (kHz)

Amplitude

(dB)

Amplitude

0.0

0.06 0.08

Time (s)

Frequency (kHz)

Amplitude

Time (s)

Figure 3. Acoustic repertoire of the advertisement call of Ate/opus laetissimus. Conventional pulsed call (A), unpulsed short call

(B), partially pulsed short call (C), pulsed short call (D), partially pulsed short call before pulsed call (E), and partially pulsed short

call within pulsed call (F). The corresponding author will provide tables of raw data for individual specimens on request.

Amphib. Reptile Conserv.

February 2020 | Volume 14 | Number 1 | e221

Granda-Rodriguez et al.

Table 2. Movement patterns of six Ate/opus laetissimus individuals.

Specimen Home range (m7?)

A4 Onn

A44 0.47

A49 0.33

Al 0.54

A6l 0.1

A36 0.59

Mean 0.36

SE 0.21

Minimum 0.1

Maximum 0.59

sector of the SNSM. Several studies in this zone have

reported more than 100 specimens in less than 150 h of

survey effort (Granda-Rodriguez et al. 2012; Rocha-Usu-

ga et al. 2017; Rueda-Solano et al. 2016a). Nevertheless,

at less than 2,100 m of elevation, the relative abundances

recorded for the species were significantly lower (Car-

| ces f ar

Occurrences

a Peet eh, att Piep den

oT Ce eee a kt Fy :

Recaptures (7) Distance to nocturnal sites (m)

i ae

5 2

8 3

> 0.8

10 1.8

b) Pia

6.67 FOZ

207 0.82

5 0.8

10 3

vajalino-Fernandez et al. 2008, 2013; Granda-Rodriguez

et al. 2012). In the model performed in this study, the

most suitable habitat was in the altitudinal range be-

tween 2,000 and 3,000 m. It is possible that changes in

the physical and structural characteristics of the habitat

at lower altitudes contribute to the decrease in either the

Habitat loss

@ Before 2000

Suitability

al Deforestation 2000-2017

IUCN criteria (B)

MM 01-02

MM 02-03

|] 03-04

[ _]04-05

| |05-06

Colombia

i o9-1

Fig. 4. Habitat suitability estimate (upper left panel), minimum convex polygon of extent of occurrence (EEO, upper right panel),

and area of occupation (AOO, lower left panel) of Ate/opus /aetissimus. The total deforested area for the analyzed period and species

occurrence locations are provided in red.

Amphib. Reptile Conserv.

35 February 2020 | Volume 14 | Number 1 | e221

Natural history of Harlequin Toad, Atelopus /aetissimus in Colombia

Habitat loss (Ha) @

2005 2010 2015

Fig. 5. Estimated annual habitat loss for Ate/opus laetissimus

in the last decade in the potential distribution (A), area of

occurrence (AOO, B), and extent of occurrence (EOO, C).

occupation or detection of A. /aetissimus.

Populations of A. /aetissimus appear to be highly dis-

proportionate in males. The first publication that men-

tioned this observation (Rocha-Usuga et al. 2017) did not

describe how the males were differentiated from the fe-

males. A sexual proportion that is biased to males can be

related to differences in the mortality rate by sex. How-

ever, it is also possible that some individuals categorized

as males corresponded to small females, especially since

the sexuality was assumed based on size (e.g., GOmez-

Hoyos et al. 2017). This is an important detail that has

been addressed only rarely in population studies of At-

elopus (Gomez-Hoyos et al. 2014; Gonzalez-Maya et al.

2018), but it is important due to the conservation interest

in the species of this genus.

Regarding population density, there is no previously

published information for A. /aetissimus. Since this study

provides the first estimation of this population attribute,

it is not possible to estimate variations among different

populations of this species. However, comparing the

population density of A. /aetissimus obtained here with

lowland species, the densities obtained with distance-

based models for Atelopus spurrelli Boulenger, 1914 and

Atelopus elegans (Boulenger, 1882) were slightly lower

(0.03 and 0.01 ind/m’, respectively, GOmez-Hoyos et al.

2014, 2017). On the other hand, the observed density of

Atelopus hoogmoedi (0.47 ind/m’, Luger et al. 2009) was

higher than the mean density of A. /aetissimus found in

this study.

Some species, such as Atelopus ignescens (Cornalia,

1849) and A. varius, had dense populations before severe

population declines, with reports of 0.025—0.75 ind/m?

Amphib. Reptile Conserv.

and 0.065—0.755 ind/m?, respectively (Ron et al. 2003;

La Marca et al. 2005). Atelopus cruciger 1s one of the

few species with information on population density after

a decline, which was 0.005—0.057 ind/m* (Lampo et al.

2012). Populations of A. cruciger had a high prevalence

of chytrid fungus, but remained stable because of the

high recruitment rate of healthy individuals in the popu-

lation (Lampo et al. 2017).

Habitat Selection and Movement Patterns

The differential use of several substrates by A. /aetissi-

mus has been previously reported, where leaf-litter and

rocks were mainly used (Granda-Rodriguez et al. 2008b).

The data reported here reinforce these findings, suggest-

ing that this species selects the most available substrates.

The structural complexity of the riparian forest occupied

by A. /aetissimus can influence its differential pattern

of habitat use, as has been described for some anurans

from southeastern Asia (Gillespie et al. 2004). Habitat

selection allows organisms to avoid adverse environmen-

tal conditions, like extremely low temperatures (Navas

1996). Recently, A. /aetissimus has been described as a

thermoconforming species, showing a direct relation-

ship between the temperature of substrate and the activ-

ity temperature (Rueda-Solano et al. 2016b). Therefore,

the differential selection of substrates could be associ-

ated with some thermoregulatory strategy. The results of

these surveys show that A. /aetissimus exhibits relatively

high nocturnal site fidelity, although the recapture rate

was decreasing gradually, probably due to the manipula-

tion of the specimens in each recapture. Recently, Rueda-

Solano and Warketin (2016) reported that A. /aetissimus

use the nocturnal sites for predatory activities, guided by

the vibration of the substrate (leaves and ferns), suggest-

ing that the use of a nocturnal perch is not exclusively

for rest.

Regarding the home range of Afe/opus, some species

such as A. carbonerensis and A. hoogmoedi possess a

mean home range much larger than A. /aetissimus (41 m?

and 38.1 + 17.7 m?, respectively; Dole and Durant 1974;

Luger et al. 2009), which could be the result of seasonal

variation. The results here indicate that individuals of A.

laetissimus can remain, at least for a short period, near to

the stream defending their territories.

Advertisement Call

The pulsed call is the most commonly known vocaliza-

tion in harlequin toads, being present in at least 17 spe-

cies (Asquith and Altig 1989; Cocroft et al. 1990; Ibafiez

et al. 1995; Jaslow 1979; Lescure 1981; Lotters et al.

1999, 2002; this study). The pulsed call of A. /aetissimus

consists of a short series of pulses (7-33 pulses) emitted

rapidly, which is remarkably different from the pulsed

calls of A. barbotini Lescure, 1981 (41-53 pulses per call,

30.35-—33.97 pulses/s, 2,000—3,000 Hz; Lescure 1981),

February 2020 | Volume 14 | Number 1 | e221

Granda-Rodriguez et al.

A. flavescens Dumeril and Bibron, 1841 (45-58 pulses

per call, 29.76—34.78 pulses/s, 2,500—3,000 Hz; Lescure

1981), A. franciscus Lescure, 1974 (31-39 pulses per

call, 22.97—23.78 pulses/s, 2,300-3,000 Hz; Lescure

1981), A. hoogmoedi (40-42 pulses per call, 33.61—35

pulses/s, 2,300—3,000 Hz; Lescure 1981), A. spumarius

Cope, 1871 (20-37 pulses per call, 38.55—45.96 pulses/s,

3,600—-4,400 Hz; Asquith and Altig 1987; Lescure 1981)

or A. reticulatus Lotters, Haas, Schick, and Bohme, 2002

(27-32 pulses per call, 75—76 pulses/s, 3,282 Hz; Lotters

et al. 2002) by having a higher number of pulses repli-

cated more quickly at a lower dominant frequency. Like-

wise, it differs from the pulsed call of A. zeteki Dunn,

1933 (42-52 pulses per call, 115-146 pulses/s, 1,381-

1,510 Hz; Cocroft et al. 1990), by having fewer pulses

replicated more quickly at a higher dominant frequency.

Other species such as A. cruciger (84—99 pulses per

call, 2,400—2,870 Hz; Cocroft et al. 1990), A. limosus

Ibafiez, Jaramillo, and Solis, 1995 (31-45 pulses per

call, 146.4-156.3 pulses/s, 2,600—2,800 Hz; Ibafiez et

al. 1995), and A. varius (43-56 pulses per call, 119-123

pulses/s, 1,750—1,965 Hz; Cocroft et al. 1990) exhibit

pulsed calls with higher numbers of pulses emitted at

considerably faster rates than A. /aetissimus. On the oth-

er hand, the structure of pulsed calls of A. /aetissimus 1s

very similar to the calls of A. chiriquiensis Shreve, 1936

(18—33 pulses per call, 59.5—82.3 pulses/s, 2,000—2,700

Hz, Jaslow 1979), A. exiguus (Boettger, 1892) [19-21

pulses per call, 2,150—2,700 Hz, Coloma et al. 2000],

A. minutulus Ruiz-Carranza, Hernandez-Camacho, and

Ardila-Robayo, 1988, (14—21 pulses per call, 59.5—67.9

pulses/s, 2,700—3,150 Hz, Cocroft et al. 1990), A. nice-

fori Rivero, 1963 (21-24 pulses per call, 53.9-65.7

pulses/s, 2,630—2,871 Hz, Cocroft et al. 1990), A. senex

Taylor, 1952 (30-34 pulses per call, Cocroft et al. 1990),

and A. tricolor Boulenger, 1902 (16-19 pulses per call,

2,970-3,450 Hz, Lotters et al. 1999). Pulsed calls of

these species also consist of shorter calls emitted at faster

rates, but with higher frequencies than A. /aetissimus in

all cases.

The second type of vocalization (short calls) has

been described for 12 species (Carvajalino-Fernandez et

al. 2017; Ibafiez et al. 1995; Jaslow 1979; Lotters et al.

1999, 2002; this study). This call is the more variable

of the two in terms of structure, by the definitions pro-

posed by Cocroft et al. (1990), which includes several

vocalizations emitted in different social contexts. Both

pulsed and short calls of A. /aetissimus corresponded to

the advertisement call context (sensu Wells 2007). This

is probably the same situation for A. varius, whose short

calls were obtained in the field, without apparent interac-

tion among individuals (Cocroft et al. 1990). Short calls

recorded in captivity for A. cruciger and A. spumarius

also can be related to advertisement calls. This call is also

reported in male-female interactions, in an amplectant

couple of A. zeteki. Wells (2007) described this type of

interaction as courtship calls. Encounter calls were re-

Amphib. Reptile Conserv.

corded in a male-male aggressive interaction in A. chiri-

quiensis (Jaslow 1979). Nevertheless, most of the short

calls described were release calls obtained at the moment

of specimen manipulation (A. chiriquiensis, Jaslow 1979;

A. limosus, Ibafiez et al. 1995; A. nahumae, Carvajalino-

Fernandez et al. 2017; A. peruensis and_A. tricolor, Lot-

ters et al. 1999). In addition, short calls of A. tricolor

cannot be included in any of these categories, since the

context of the recording was not described clearly (Lot-

ters et al. 2002). Previously, the role of vocalizations in

the communication of the genus Ate/opus has been dis-

torted by the absence of several elements of the auditory

apparatus (McDiarmid 1971) and the conspicuousness of

its visual communication (Jaslow 1979; Crump 1988).

Nevertheless, the complexity and diversity of vocaliza-

tions described and reviewed here suggests that their

roles in communication may be underestimated.

Potential Distribution and Habitat Loss

According to IUCN SSC Amphibian Specialist Group

(2014), A. laetissimus is a species restricted to the moist

low montane forest life zone, at altitudes between 1,500-—

2,880 and an area of 797 km?. In this study, localities

are reported between 900 and 2,880 m asl and an EOO

of 1,074 km/?, higher than that reported by IUCN. De-

spite this increase in the distribution of the species which

could be associated with new samplings, we recommend

its status of Endangered (EN) be maintained. The niche

model of A. /aetissimus suggests that the potential distri-

bution is restricted to forests in humid zones at the north-

western and northern flanks of the SNSM. The results

show that the AOO of the species may not exceed 52 km”.

The analysis by the model suggested the environmental

layers that most influenced the distribution of the species

are the average temperature of the coldest quarter, the

elevation, and the human footprint. In the case of am-

phibians, important influences of the climate on their dis-

tribution have been described, therefore, factors such as

climate change could significantly alter populations of A.

laetissimus. Among the potential changes are alterations

of the precipitation and temperature regimes, resulting

in an increase in the annual temperature ranges and thus

affecting habitat quality and the availability of specific

resources for A. /aetissimus (Zhang and Yan 2014). On

the other hand, human influence generates a very marked

negative effect on this type of species in terms of degra-

dation and loss of habitat (Grant et al. 2016).

The model performed here represents the first empiri-

cal estimation of the distribution of this species based on

distribution modeling, and it also uses the largest compi-

lation of localities. Additionally, recent samples in areas

with high suitability predicted by the model support the

reliability of the prediction (Rueda and Warkentin 2016).

In any case, the predictions of these models should be in-

terpreted with caution and they should be considered as a

first approximation to the real distribution of the species,

February 2020 | Volume 14 | Number 1 | e221

Natural history of Harlequin Toad, Atelopus /aetissimus in Colombia

helping to focus the sampling efforts in order to further

adjust and refine the predictions of the distribution of this

species in the future.

Although the habitat loss identified does not represent

a high percentage in relation to the total habitat of the

species, the trends showed a significant increase in loss in

recent years (Ribeiro et al. 2018). However, considering

that Hansen et al. (2013) does not differentiate between

types of vegetation (natural and exotic plantations), the

results could vary. This is important since A. /aetissimus

has scarcely been associated with exotic plantations of

Pinus spp., or in streams associated with this type of cov-

erage in the locality of San Lorenzo (Granda-Rodriguez

et al. 2012). In addition, it is also not known how coffee

and avocado plantations can affect this species, since in

the middle- and upper-part of the distribution large areas

of these crops are present (Fundacion Pro-Sierra Nevada

de Santa Marta 2000). It is necessary to carry out studies

at smaller scales in the distribution area of A. /aetissimus,

allowing the identification of the landscape dynamics of

forest patch isolations, connectivity, and the different

elements that may have negative consequences for this

species (Palmeirim et al. 2018). In this sense, remaining

remnants of forest become important for the maintenance

of Atelopus laetissimus, as well as other endemic spe-

cies of the SNSM. However, in the southern sector of the

SNSM there is high fragmentation and habitat degrada-

tion; and this site is (was?) inhabited by A. arsyecue and

A. walker, species that have not been seen in the field for

more than 20 years, providing possible evidence for the

risk of disappearance of this type of toad.

Acknowledgements.—We are grateful to the people from

Minca village and Unidad de Parques Naturales. We

thank Vanesa Pacheco, Adolfo del Portillo, Miguel De

Luque, and Liliana Saboya for their assistance during the

field work. We thank Cristian Estades, Jaime Hernandez,

and the anonymous reviewers for their valuable com-

ments on the manuscript.

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Hernan Granda Rodriguez is a Biologist from the University of Magdalena, Santa Marta, Colombia,

who obtained his Master’s degree in Wild Areas and Nature Conservation from the University of Chile,

Santiago, Chile. Hernan is currently a professor in the environmental engineering program at the University

of Cundinamarca, Cundinamarca, Colombia. His research interests focus on the conservation of amphibians

and reptiles, from the perspectives of both biological issues and social issues, such as the public's perception

of herpetofauna.

. Andrés Camilo Montes-Correa is a Colombian young biologist and herpetologist (in training) at the Uni-

versidad del Magdalena (Santa Marta, Colombia). His research interests include the taxonomy, systematics,

and ecology of the Colombian herpetofauna, especially lizards and freshwater turtles. Andrés has conducted

investigations on habitat use by freshwater turtles of the Colombian Llanos (for his B.Sc. thesis); ecology,

taxonomy, and distribution of small geckos and cryptic dendrobatid frogs; and synecological studies of

herpetofauna. His current project is on the taxonomy, ecology, distribution, and conservation of Lepido-

blepharis miyatai, a small and endangered gecko endemic to the coastal mountains (Sierra Nevada de Santa

“ Marta) of the Colombian Caribbean.

Juan David Jiménez-Bolaiio is a wildlife photographer and has a B.Sc. in Biology from the University of

Magdalena (Santa Marta, Magdalena, Colombia). Juan’s work has focused mainly on the ecological and

diversity patterns of Neotropical herpetofauna. Currently, Juan is interested in studying diversity patterns in

high Andean environments.

Alberto J. Alaniz, Geographer, B.Sc., M.Sc. His work is focused on ecological modelling applied to

biological conservation and epidemiology, and his recent research has been on the development of spatially

explicit infection risk models for Zika virus at global and local scales. Currently, Alberto is a researcher

in the Ecology and Conservation Lab in the Technological Faculty at the University of Chile, and he is a

founding member of the Center for Spatial Ecology and Environment (Ecogeografia), located in the city of

. Santiago, Chile.

Patricio Hernaez is a Marine Biologist by training at the Universidad Arturo Prat (Chile), with a Master’s

degree in Biology from the Universidad de Costa Rica (Costa Rica), and a Ph.D. in Biological Sciences from

the Universidade de Sao Paulo (Brazil). His research focus is divided into three main lines: (1) Taxonomy

of recent groups with an emphasis on Crustacea, (11) Population dynamics, and (ili) Fisheries. Patricio

is currently a researcher associated with the Centro de Investigaciones Marinas y Limnologicas of the

Universidad de Tarapaca, Chile, and the Grupo de Pesquisa em Biologia de Crustaceos of the Universidade

Estadual Paulista ‘Julio de Mesquita,’ Brazil.

Pedro E. Cattan has a D.V.M. and a Ph.D. in Biological Sciences from the University of Chile, and he

was the Chairman of the Department of Biological Sciences of the University of Chile for 10 years. Pedro

has developed two main lines of research, one on the study of populations in wild vertebrates and the other

on disease ecology at the wildlife level. He currently directs projects on the vectors and wild reservoirs of

Chagas disease.

Amphib. Reptile Conserv. 42 February 2020 | Volume 14 | Number 1 | e221

Official journal website:

amphibian-reptile-conservation.org

Amphibian & Reptile Conservation

14(1) [General Section]: 43-54 (e222).

Do growth rate and survival differ between undisturbed and

disturbed environments for Sceloporus spinosus Wiegmann,

1828 (Squamata: Phrynosomatidae) from Oaxaca, Mexico?

‘Carlos A. Torres Barragan, ?Uriel Hernandez Salinas, and **Aurelio Ramirez-Bautista

‘Instituto Politécnico Nacional, Centro Interdisciplinario de Investigacion para el Desarrollo Integral Regional (CIIDIR) Unidad Oaxaca, Hornos

No. 1003, Col. Noche Buena, Santa Cruz Xoxocotlan, Oaxaca, Oaxaca 71230, MEXICO ?Instituto Politécnico Nacional, Centro Interdisciplinario

de Investigacion para el Desarrollo Integral Regional (CIIDIR) Unidad Durango, Calle Sigma 119 Fraccionamiento 20 de Noviembre II, Durango,

Durango 34220, MEXICO ?Laboratorio de Ecologia de Poblaciones, Centro de Investigaciones Bioldgicas, Instituto de Ciencias Bdsicas

e Ingenieria, Universidad Autonoma del Estado de Hidalgo, Km 4.5 carretera Pachuca-Tulancingo, 42184, Mineral de La Reforma, Hidalgo,

MEXICO

Abstract.—Demography is intimately related to the evolution of the life history of a species, since it describes

the patterns of variation in the growth, maturation, reproduction, and survival of an organism through

populations, species, and environments. In this study the growth, survivorship, and population structure were

evaluated for an oviparous lizard, Sceloporus spinosus from two sites, a relatively undisturbed area (UA) and

a disturbed area (DA; zone of land-use change) within the Natural Protected Area Yagul of southern Oaxaca,

Mexico. The results showed different relative densities between seasons (higher during the wet season than

the dry season), but not between populations. Males and females from the UA and DA showed similar growth

rate patterns, and both sexes reached sexual maturity at a similar body size. The highest survival rates and

recapture probabilities were found in the UA; however, males from both populations showed higher survival

rates than females. Overall, this study suggests that land-use changes do not seem to cause wide variation

in the analyzed demographic characteristics of this species. This work describes and quantifies demographic

effects on some life history characteristics of aspecies endemic to Mexico. We argue for the need to analyze and

compare many capture-recapture data for a species between locations in order to obtain a better assessment

of the variation in the life history characteristics analyzed.

Keywords. Age class, Cormack-Jolly-Seber model, demography, density, natural protected area, toe-clipping method

Citation: Torres Barragan CA, Hernandez Salinas U, Ramirez-Bautista A. 2020. Do growth rate and survival differ between undisturbed and disturbed

environments for Sceloporus spinosus Wiegmann, 1828 (Squamata: Phrynosomatidae) from Oaxaca, Mexico? Amphibian & Reptile Conservation

14(1) [General Section]: 43-54 (e222).

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: 7 August 2018; Accepted: 21 October 2019; Published: 18 February 2020

Introduction characteristics of the environments that they inhabit (e.g.,

pristine or fragmented). Therefore, the conservation

The variation in life history characteristics of lizards is _ priority of populations of a widely distributed species, is

considered to be an outcome of phenotypic plasticity

driven by changing environmental conditions (Stearns

1992; Adolph and Porter 1996). Studies investigating

demographic parameters (e.g., density, sex ratio, natality,

growth rate, age classes) and life history (e.g., SVL at

sexual maturity, survival, reproduction, fecundity) in

lizards have shown that different life strategies (e.g.,

growth rate, survival) in these vertebrates have evolved

due to environmental changes caused by habitat loss and

by changes in land use (Dunham 1982; Stearns 1992).

This implies that the populations of any species may

evolve different life history strategies according to the

to assess the tolerance of their life history characteristics

to certain environmental factors (precipitation, humidity,

radiation, pollution, deforestation, and others) that occur

throughout the distribution of the species; therefore, this

is a feasible method to test for changes in their fitness

(Walkup et al. 2017). Evolution within and across species

that inhabit fluctuating environments has resulted in

changes of their life history strategies, such as size and

age at sexual maturity, fecundity (clutch size), growth

rate, and survival (Stearns 1992); and these changes

have been found within different populations of a single

species that is widely distributed (Dunham 1982; Cruz

Correspondence. ! augusto.torres007@gmail.com, ? uhernndez3@gmail.com, **ramibautistaa@gmail.com

Amphib. Reptile Conserv.

February 2020 | Volume 14 | Number 1 | e222

Sceloporus spinosus in Oaxaca, Mexico

et al. 2014; Pérez-Mendoza et al. 2014; Cruz-Elizalde

and Ramirez-Bautista 2016). These variations in life

history characteristics have been documented in several

species that inhabit environments with different degrees

of disturbance (Cruz et al. 2014; Cruz-Elizalde and

Ramirez-Bautista 2016; Walkup et al. 2017). However,

strong fragmentation of habitats by land use change,

pollution, and global warming have adverse effects on

these strategies, that consequently lead to population

decline at the local level (Sinervo et al. 2010).

Due to concerns regarding the effects of environmental

disturbances, some _ researchers have generated

conservation models for various biological groups

(e.g., birds, Escalante et al. 1998; mammals, Ceballos

and Oliva 2005) that include significant amounts

of information on the natural history of individual

species. For example, Sinervo et al. (2010) described

patterns of species decline and extinction in a diverse

assemblage of lizard species of genus Sce/oporus under

thermoregulatory stress induced by global warming.

Chavez (2011) and Calder6n-Mandujano (2011) noted

that land use change is another factor that has resulted

in high levels of population decimation and extinction

among lizards and amphibians. However, each species

responds in different ways according to the pressures of

their local environment (Tews et al. 2004; Suazo-Ortufio

et al. 2007). This pattern has been documented in several

wide-ranging species that occur in relatively pristine

habitats as well as sites that are subject to varying

degrees of disturbance (e.g., Sceloporus grammicus,

Pérez-Mendoza et al. 2014; S. minor, Garcia-Rosales et

al. 2017; and S. variabilis, Cruz-Elizalde and Ramirez-

Bautista 2016). Therefore, considering these factors,

herein, the effects of a pristine and a disturbed habitat

on some life history characteristics were evaluated in

two populations of Sce/oporus spinosus (Eastern Spiny

Lizard) in southeastern Mexico.

Sceloporus spinosus 1s a species endemic to Mexico,

and adults are of medium body size for the genus (120

mm snout-vent length, SVL; Ramirez-Bautista et al.

2014). The scales of the body are strongly keeled and

mucronate. This species feeds on insects and other

invertebrates, and it 1s oviparous with a clutch size of

eight to 31 eggs and a mean of 18.5 (Valdéz-Gonzalez

and Ramirez-Bautista 2002). This lizard is found from

Durango to Oaxaca, and inhabits arboreal and saxicolous

landscapes (Torres-Barragan 2015) in both temperate

and semiarid regions, at an elevation range from 1,900 to

2,700 m (Canseco-Marquez and Gutiérrez-Mayén 2010).

In Yagul Natural Protected Area (NPA), this species is

distributed in a mountain range with elevations from

1,600 to 2,000 m. So far, there is limited information

regarding demographic aspects of this species in pristine

and disturbed areas such as Yagul NPA. In this framework,

the goal of this study was to compare and assess key

demographic characteristics, such as density, growth,

survival, and population structure of S. spinosus in two

Amphib. Reptile Conserv.

contrasting environments, in Oaxaca, Mexico. Therefore,

considering that land use change promotes variation in

demographic characteristics and life histories in various

vertebrate groups (Adolph and Porter 1996; Flatt and

Heyland 2011), these demographic characteristics were

expected to differ as a function of the environments

where each population of Sceloporus spinosus occurred.

Materials and Methods

Study Area

This study was carried out at two sites in Yagul Natural

Protected Area in the municipality of Tlacolula de

Matamoros, Oaxaca, Mexico (Fig. 1). The municipality

encompasses 1,076 ha, ranging in elevation from 600

to 2,500 m. The vegetation at the site is represented by

tropical dry forest; however, much of it has been replaced

by crops and grazing areas. The climate is semi-warm,

with temperatures ranging from 16 °C to 26 °C, and mean

annual precipitation from 400 to 800 mm (INEGI 2005).

Two sites of 1 ha each were chosen for this study.

The first site was considered the undisturbed area (UA;

16.957922 N, -96.429953 W; 1,800 m), with a vegetation

cover of 83% and an arboreal density of 697 individual

trees/ha. This cover includes 80% tropical dry forest, 10%

flood zone, 3% reedbed (Arundo donax), and 2% surface

without vegetation (Torres-Barragan 2015). The second

site was a disturbed area (DA; 16.959617 N, -96.450633

W; 1,652 m; Fig. 1). This site is an open area with agave

plant cultivation and extensive grazing areas; canopy

cover 1s 1% with an arboreal density of eight individual

trees/ha; 50% of the land is used for cultivation of agave

plants (Agave angustifolia), 20% for induced pasture,

10% for living fences (Prosopis, Yucca, Celtis, Acacia,

Opuntia, and Schinus), and 20% of the surface has no

vegetation (exposed floor; Torres-Barragan 2015).

Data Collection

Twelve sampling events were conducted at each site

(UA and DA) from January 2014 to January 2015. Each

sampling event was carried out over a single three-day

period in each month at each of the two sites (for a total

of six sampling days per month), with a sampling effort

of three people from 0900-1800 h.

The method of mark-recapture of Lemos-Espinal

and Ballinger (1995) and Ramirez-Bautista (1995) was

used in this study. This method consisted of ectomization

of phalanges (e.g., toe-clipping), a permanent marking

technique that makes it possible to recognize every

previously marked individual during each subsequent

sampling event. Toe-clipping is commonly used to follow

cohorts of lizard populations (Dunham 1978; Tinkle

1961, 1969). More recently, Guimaraes et al. (2014) and

Olivera-Tlahuel et al. (2017) expressed some concern

when using this method due to observed effects on the

February 2020 | Volume 14 | Number 1 | e222

Torres Barragan et al.

1877000

Tlacolula de

Matamoros

1875000 1876000

1874000

)

Salado River

770000

771000

772000

SYMBOLOGY

MUA

[DA

—— Watercourses

[]} Polygon MNY

1876000

Level curve

1875000

~—-—= Road

LC] Urban area

ee Archaeological Area Yagul

1874000

773000

Scale 1:27804

Fig. 1. Map of the study area. The green polygon depicts Yagul Natural Protected Area, including the two sampling sites (UA =

undisturbed area; DA = disturbed area, land use change).

behavior and health of some lizard species, and therefore,

on survival. Although toe-clipping could affect survival,

lizards at both sites were toe-clipped similarly, so any

negative bias in survival estimates should apply equally

to both sites. The SVL of each lizard was measured with

a digital caliper (to the nearest 0.01 mm), and body mass

with a balance (+ 0.01 g).

Relative Density and Population Structure

Based on the number of captured and recaptured

individuals from both populations during the study,

the relative density of each population was determined

using the equation, N = M/R, where N = number

of unknown individuals in the population; M = the

number of marked individuals; and R = the number

of recaptured individuals/surface area. To determine

the population structure for each site, size classes

(SVL) were determined based on those used by Leyte-

Manrique et al. (2017) with Sceloporus grammicus.

These authors related the SVL of each age category

based on anatomical traits, yielding classification

categories of: offspring (SVL < 48 mm), juveniles

(49-69 mm), and adults (females and males > 70 mm).

Females were considered to be adults if they contained

eggs in the oviduct, which were identified by palpation

of the ventral region (Galan 1997). Whereas males were

considered to be adults when they showed the bulky

tail base indicative of sperm production (Lozano et

al. 2014). Relative densities of lizards were compared

Amphib. Reptile Conserv.

between locations and seasons by means of a Student’s

t-test (Zar 2014).

Growth Rate

Growth rates were assessed for males and females from

each population by considering only those lizards with

recapture intervals greater than 30 and less than 100 days.

Therefore, growth rate was estimated with the formula:

GR = (SVL, — SVL,)/days, where growth rate (GR) is

the difference in recorded SVL between the last recapture

(SVL,) and first capture (SVL,) divided by the number

of days that had elapsed (Dunham 1978; Zamora-A brego

et al. 2012). Then, nonlinear regression models of Von

Bertalanffy, logistic by body size (SVL), and logistic

by body mass were used (Dunham 1978); and growth

rates for both sexes and populations were compared.

The first model (Von Bertalanffy) describes a pattern in

which smaller individuals (in SVL) show faster growth

rates than larger ones (Dunham 1978; Zamora-Abrego

et al. 2012). In contrast, the logistic models predict

that individuals smaller in SVL will grow moderately

faster to reach intermediate sizes, and after reaching

their maximum growth rate, that rate will decrease in

a non-linear direction as size increases (Dunham 1978;

Schoener and Schoener 1978; Zamora-Abrego et al.

2012). The difference between the two models 1s that the

maximum growth rate attained under a logistic by length

(SVL) model is observed at early ages, while maximum

growth rate under a logistic by body mass model will

February 2020 | Volume 14 | Number 1 | e222

Sceloporus spinosus in Oaxaca, Mexico

be observed at later ages (Dunham 1978). Detailed

descriptions of each model can be reviewed in Dunham

(1978) and Schoener and Schoener (1978). Selection of

the best model was based on the best fit to the observed

growth rates for both sexes, chosen by the lowest value

of the residual mean square (RMS) and highest values of

coefficients of determination or correlation (R*?; Dunham

1978; Schoener and Schoener 1978).

Once selected, the models were developed with

confidence intervals following Schoener and Schoener

(1978) for the growth parameter (r), and the asymptotic

(A,) was calculated by the formula:

@,— JRF, ani S} = Bj = 9, + JRF, en S}

where B, is the adjusted parameter j, 0, is the parameter

estimated B., S, 1s the asymptotic standard deviation of

bees _ {9 is the value F _, of a tail with & and N —k

degrees of freedom, N is sample size, and k 1s the number

of adjusted parameters. These confidence intervals

provide maximum reliability for each parameter (r and

A,) and are considered significantly different between

sexes if they do not overlap (Schoener and Schoener

1978). On the other hand, the residuals of the growth rate

(removal of effect size) were used to determine by two-

way ANOVA if there were differences in the patterns of

growth rates between factors (sexes and populations).

The residuals are the result of the relationship between

the SVL and the growth rate under the model with the

best fit (Schoener and Schoener 1978). Finally, based on

the values of the growth parameter (r) and asymptotic

(A,) obtained from the best-fit model, together with the

average values of SVL of offsprings at hatching (LO = 42

mm in SVL), the ages in days were determined for both

males and females at which they reach sexual maturity

(Dunham 1978; Schoener and Schoener 1978; Zamora-

Abrego et al. 2012). The growth models for both sexes

and populations were developed with Statistica program,

version 7.0.

Estimates of the Survival Models

Captures and recaptures of marked individuals

allowed estimates of demographic parameters, such

as survival (@) and recapture (p); and both parameters

were estimated from different models that represent

distinct biological hypotheses of survival (Lebreton et

al. 1992). Both and p can be constant (c) over time

or vary as a function of time (¢) and between sex, and

for their assessment a general model was considered

which allowed the determination of whether survival

rates and recaptures were different between sexes

and populations. This model is: @ (groups [males and

females in two populations = four groups]*time) p

(groups*time), and it calculates the probability that

survival and rate of recapture are different between

groups over different periods of time.

Amphib. Reptile Conserv.

For developing the different models based on encounter

histories of each individual (e.g., 100101), zero represents

sampling when a lizard was not seen in the area, while

1 represents those sampling events when lizards were

marked and recaptured (Lebreton et al. 1992). Encounter

histories were analyzed and modeled with software Mark

6.0 (White and Burnham 1999) using the subprogram

“only recaptures” with the goal of obtaining estimates of

survival and recapture rates grounded in the techniques

of maximum likelihood under the model developed by

Cormack-Jolly-Seber (Lebreton et al. 1992). The model

that showed the best fit to the capture-recapture data

was the one with the lower Akaike information criterion

(AIC) value; however, when there was a difference of 2

between A/C values of the two models, both models were

assumed to have approximately the same fit to the data

(Burnham and Anderson 2002). Survival and recapture

results are represented with confidence intervals of 95%.

Results

Relative Density and Population Structure

In the surveys, 271 individuals were marked across

both sites (UA = 149 and DA = 122): of these, 113 were

recaptured (73 in UA and 40 in DA; Table 1). In some

cases, several individuals were captured as juveniles and

then recaptured as adults; but most of the recaptures were

adult males and females (Table 1). The relative density

between seasons was different (oN = -2.023, P = 0.05;

wet: 6.95 + 0.41 [4.92-10.07]; dry: 9.70 + 1.29 [2.68—

17.21]), but not between populations (¢,,, = -0.116, P

= 0.98; UA: 8.33 + 0.97 [5.68-16.11]; DA: 8.33 + 1.11

[3.28-17.21]). The relative density for UA (both sexes

and all age classes) was 149 individuals/ha, whereas for

DA it was 122 individuals/ha. In UA, offspring emerged

from July to December, but the peak hatching period was

in September; juveniles were recorded from October to

April, but the highest population of this age class was in

December; adults were present throughout the year, but

the highest numbers of captures were in April and May

(Table 1). Offspring from DA were found from August

to November, with peak density in August; juveniles

appeared from October to March, with density peaking in

December; and adults were seen from September to July,

with the greatest densities from April to May (Table 1).

Growth Rate

The length logistic model showed the best fit to the

growth rate data for males and females from UA; in

contrast, the Von Bertalanffy model showed the best fit

to the growth rate data for both males and females of DA

(Fig. 2 and Table 2). The logistic model by length showed

that growth rates for males (r + EE: 0.007 + 0.0005) and

females (r + EE: 0.008 + 0.0008) from UA were similar,

whereas the asymptotic growth curve of females (A, =

February 2020 | Volume 14 | Number 1 | e222

Torres Barragan et al.

Table 1. Numbers of Sce/oporus spinosus individuals in each age class during each study month at Yagul Natural Protected Area.

UA = Undisturbed area, DA = Disturbed area. * Indicates the rainy months. A = adult, J = juvenile.

UA Offspring Juveniles Adults

Samples Males Females Males Females Males Females Recaptures

January 0 0 1 2 1 0 0

February 0 0) is) 4 2 1 19A

March 0 0 2 4 7 2 3GA, 19, 295

April 0 0) 3 3 9 9 ASA, 52,28)

May 0) 0 0) 0) 7 10 AGA, SQA

June* 0) 0) 0) 0) 2 6 IGA, SOA

July* 0) 1 0) 0 7 4 SSA, 29A

August* 3 1 0 0 2 3 29,14)

September* i) 2 0) 0 5 1 30'A, 129A, 23)

October a 1 0) 1 4 3 3GA, 29A, 28, 19)

November 4 0) 3 2 6 0) SSA, 28, 19)

December 1 0) 7 2 2 0) SSA, 3Q9A

Density (#/area) 0.0019 0.0037 0.0093

Total 14 5 19 18 54 39 73

DA Offspring Juveniles Adults

Samples Males Females Males Females Males Females

January 0 l 2 2 2 2 0

February 0) 0 1 0 2 1 0

March 0 0 2 2 5 3 IGA

April 0) 0) 0) 0 10 6 6GA, 32A

May 0) 0 0) 0 11 10 AGA, SQA

June* 0) 0) 0) 0 1 5 29A,1dA

July* 0) 0 0) 0) 4 5 SSA, 29A

August* 3 4 0) 0) 0 0 0

September* 1 2 0 0 1 3 0

October 1 0 2 4 0) 2 19J,29A

November 1 0 4 2 0 2 19A, 3d)

December 0) 0 2) 6 1 1 IGA, 19, 295

Density (#/area) 0.0013 0.0032 0.0077

Total 6 yi 16 16 37 40 40

EE: 105.359 + 3.225 mm) was slightly higher than that

of males (A, + EE: 101.706 + 1.587 mm; Table 2). On

the other hand, for males and females from DA, the Von

Bertalanffy model showed that male (r + EE: 0.005 +

0.0008) and female (r+ EE: 0.005 + 0.0011) growth rates

were similar (Table 2); however, the females reached an

asymptotic size (maximum size) that was slightly larger

(A, + EE: 108.058 + 4.139 mm) than the males (A, =

EE: 103.253 + 3.723 mm; Table 2). Average values of

the residuals of growth rates for males and females from

UA, obtained with both the logistic by length model and

a two-way ANOVA, did not show significant differences

(males: -0.001 + 0.009; females: 0.003 + 0.019) between

dry (F’,, = 0.047, P = 0.8300) and wet seasons (0.001 +

1,29

Amphib. Reptile Conserv.

0.010, -0.004 + 0.011, respectively), between sexes (F, ,,

= 0.003, P = 0.9577), or interactions between factors

(season*sex; F, ,, = 0.116, P = 0.7358). A similar pattern

occurred for DA, where no differences were found in

males (-0.003 + 0.013) and females (-0.007 + 0.015)

between dry and wet seasons (Pag = 0.975, P = 0.3333;

males = 0.021 + 0.009 and females = 0.015 + 0.033),

between sexes (F, ,, = 0.044, P = 0.8358), or interactions

between factors (season*sex; Ff’ ,, = 0.004, P = 0.9472).

On the other hand, there were no differences in the

overall growth rates of males from the two populations

(fF ,, = 0.234, P = 0.6317), in neither dry (UA: -0.001+

0.009; DA: -0.003+0.013) nor wet (UA: 0.001 + 0.010;

DA: 0.021 + 0.009) seasons (F, , = 0.535, P = 0.4699);

131

February 2020 | Volume 14 | Number 1 | e222

Sceloporus spinosus in Oaxaca, Mexico

Growth rate (day/mm)

SVL (mm)

Growth rate (day/mm)

40 50 60 70 380 90 100 110 50 60 70 80 90 100 110

SVL (mm) SVL (mm)

Fig. 2. Growth rate of Sce/oporus spinosus. (A) Undisturbed area (UA) males, (B) Disturbed area (DA) males, (C) UA females, and

(D) DA females. Black circles represent data points for individual lizards. Modeled relationships between growth and body sizes of

males and females: solid lines = Von Bertalanffy, dashed lines = logistic by length, and dotted lines = logistic by mass.

—

= 240

alice

ri] -|

= 160 -

: 120

o |

= 80 |

oO |

om 40

= 0.

x DA UA

€ 0.0120 0.0120 -

O — 0.0100 J 0.0100 -

i % M—_

oD —

o rT 0.0080 0.0080 -

2 = 0.0060 0.0060 - a

g 0.0020 0.0020 -

0 0.0000 0.0000 =

DA UA DA UA

Fig. 3. Means and 95% confidence intervals of the Asymptotic growth (A,) and Characteristic growth (r) parameters obtained by

the Von Bertalanffy and logistic by length models for males and females of Sce/oporus spinosus in both Disturbed area (DA) and

Undisturbed area (UA) populations.

Amphib. Reptile Conserv. 48 February 2020 | Volume 14 | Number 1 | e222

Torres Barragan et al.

therefore, the interaction term (locality*season) was not

significant (F' ,, = 0.401, P = 0.5312). The same pattern

was found in females, as there were no differences

between localities (F’ tog OOS 0.8130), or seasons

(F fap ee Ee 0.7284); and, therefore, the interaction

term (locality*season) was also not significant (F,,, =

0.587, P=0.4519).

Growth parameters (r and A,) were similar for

males and females in UA and DA populations (Fig. 3);

confirming that there is no difference between the sexes.

In UA, the growth rate showed that males reached sexual

maturity at SVL 85 mm at age 210 d (7 months), whereas

females attained sexual maturity at SVL 89 mm at age

280 d (9 months). In DA, males reached sexual maturity

at SVL 81 mm at age 210 d, and females at SVL 85 mm

at age 280 d.

Survival Model Estimation

To analyze survival rate (@) and recapture (p) of S.

spinosus in UA and DA populations, a set of models was

developed (Table 3). The single model that described

survival rate as varying between groups (sexes) and

where the recapture rate was constant [® (sex) p(c)] was

chosen as the best fit for both populations (Table 3A).

Based on this model, the survival rate for males from

UA (0.82) was higher than that of females (0.70), while

the recapture rate was similar for both sexes (0.40; Table

4A). In addition, the survival rate for males from DA

(0.75) was higher than that of females (0.65), and the

probability of recapture was higher for males (0.40) than

for females (0.35; Table 4A). These values are lower than

those found for UA, which suggests a higher probability

of survival and recapture in UA than DA. On the other

hand, survival and recapture rates by season (wet and dry)

in UA and DA populations showed that the model with

the best fit was ® (c) p(season), indicating that survival

rate is constant, and the probability of recapture varies

between seasons (Table 3B). According to this model, the

survival rates in both populations were higher in the dry

(UA: 0.76, DA: 0.92) than the wet (UA: 0.54, DA: 0.49;

Table 4B) season.

Discussion

More lizards were marked at the UA site than the DA

site during this study. However, at both UA and DA the

greatest numbers of adult recaptures were in April and

May, due to the peak in reproductive activity during

these months (Valdéz-Gonzalez and Ramirez-Bautista

2002). According to the recapture data, lizards born in

August-September reached the minimum SVL at sexual

Table 2. Growth parameters for Sce/oporus spinosus males and females from UA and DA populations obtained from each growth

model. RMS = residual mean square, R* = coefficient of determination, A, = asymptotic of growth, r = parameter of growth, + =

standard error.

UA Model RMS

Males (n = 20)

Von Bertalanffy 0.029

Logistic by length 0.019

Logistic by weight 0.025

Females (n = 13)

Von Bertalanffy 0.029

Logistic by length 0.019

Logistic by weight 0.023

DA Model RMS

Males (” = 15)

Von Bertalanffy 0.027

Logistic by length 0.035

Logistic by weight 0.048

Females (n = 13)

Von Bertalanffy 0.030

Logistic by length 0.032

Logistic by weight 0.429

Amphib. Reptile Conserv.

R? A, r

0.730 109.480 + 5.223 0.003 + 0.0003

0.849 101.706 + 1.587 0.007 + 0.0005

0.797 100.081 + 1.114 0.012 + 0.0009

0.595 122.243 + 12.423 0.003 + 0.0007

0.736 105.359 + 3.225 0.008 + 0.0008

0.687 102 482° 2°29 1 0.012 +0.0011

R? A, r

0.809 103.253 + 3.723 0.005 + 0.0008

0.756 99.762 + 2.410 0.0099 + 0.0010

0.664 98.523 + 1.995 0.0141 + 0.0016

0.625 108.058 + 4.139 0.005 + 0.0011

0.601 100.178 + 3.225 0.008 + 0.0012

0.046 99 340 + 3.7378 0.012 + 0.0017

February 2020 | Volume 14 | Number 1 | e222

Sceloporus spinosus in Oaxaca, Mexico

Table 3. Models describing survival rate (@) and recapture (p)

of Sceloporus spinosus males and females in UA and DA with

the Jolly-Saber model using the Mark program. The models are

fitted with the Mark program considering that ® y p (probability

of survival and recapture) can be either constant (c) or varying

between sex (s), season (dry and wet), and population. AIC =

Measurement of the level of adjustment and parsimony of each

model, A = difference of AIC, W, = weight of AIC, K = number

of parameters.

A. Models for estimation of sex and populations.

Model AIC A, W, K

Dee) 231.98 0 0.69 12

® (c) p(c) 234.09 2.11 0.24 13

® (population) p(c) 238.22 “625. =:0,03" 412:

® (population) p(sex) 240.54 857 0.01 13

® (sex) p(c) 241.68 9.71 0.01 3

® (c) p(sex) 242.11 10.13 0 3

® (population) p(population) 243.39 11.42 0 21

® (sex) p(sex) 243.84 11.86 0 4

B. Models for estimation of season (dry and wet) and populations.

Model AIC A. Ww. K

Ui I

® (c) p(season) 224,02 0 0.88 3

® (season) p(c) 231.98 4.96 0.07 12

® (season) p(population) 237.17 JOA6. .0,01 3

® (c) p(c) 240.11 13.1 0 2

O (ft) p(t) 243.39 16.38 0 21

® (population) p(season) 247.87 20.86 0 2

maturity (> 80 mm) in April-May of the following year,

a similar pattern seen in another population of the same

species (Valdéz-Gonzalez and Ramirez-Bautista 2002).

Recruitment of offspring, together with adult males

and females, and a few subadults, results in population

growth with respect to these age structures. These

events are synchronized with an increase in ambient

temperatures (from 20.9 °C in April to 21.2 °C in May)

and precipitation (from 113.3 mm in June to 114.4 mm

in May) in the region, which also coincide with high

production of food in the environment (Dunham 1982;

Ramirez-Bautista and Vitt 1997).

In general, these results revealed that lizard density

was significantly similar between populations, but

not between seasons (higher in the wet season than in

the dry season). The higher density of lizards found

in the wet season could be explained by a high supply

of the food consumed by this species. However, this

assumption brings up additional questions regarding the

feeding habits of other species living in sympatry with

S. spinosus in disturbed and undisturbed environments.

Therefore, it is necessary to investigate whether those

sympatric species have the same possibilities of acquiring

available resources (food and microhabitat), or if they

Amphib. Reptile Conserv.

display different activity schedules that allow them to

obtain resources more efficiently. Such studies would

certainly expand our knowledge on the natural history of

this species, and consequently enable the development of

more effective conservation strategies.

The growth rate models used here showed that males

and females in both populations grow at the same rate

and reach maximum SVL at similar sizes. These results

were found by the logistic model by length (UA) and

Von Bertalanffy (DA), models that are known to fit most

growth analyses for at least some lizard species of the

family Phrynosomatidae (Lemos-Espinal and Ballinger

1995; Zufiga-Vega et al. 2008; Pérez-Mendoza et al.

2014; Ramirez-Bautista et al. 2016). The pattern of low

growth rate variation in both populations observed in this

study could suggest that populations of this species are

able to inhabit areas with certain degrees of disturbance,

as has been shown in other species of lizards analyzed

by D’Cruze and Kumar (2011) in both disturbed and

undisturbed environments.

On the other hand, a homologous pattern in growth

rates for males and females in both UA and DA

populations may be due to the similarities in SVL at birth

and the SVL at sexual maturity. For the former (SVL at

birth), growth rates are likely to be regulated by predation

intensity, acting mainly on offspring and juveniles of

both populations (Schoener 1979; Andrews 1982). This

interpretation could be different 1f both demographic

parameters (survivorship and recaptures) evaluated for

each site had changed based on an increased number

of recaptures. This pattern has also been observed in

populations of S. grammicus from Central Mexico

(Pérez-Mendoza et al. 2013, 2014).

The life history characteristics studied here for

this species could have significant plasticity among

its populations, and therefore, small differences in

temperature, precipitation, and food between UA and DA

would not have apparent effects (Valdez-Gonzalez and

Ramirez-Bautista 2002; Valencia-Limon et al. 2014). A

similar pattern in growth rates also occurs between sexes

and age classes in other lizard genera (e.g., Xenosaurus

spp.; Molina-Zuluaga et al. 2013).

The low variation in SVL at sexual maturity observed

in the growth curve within and between populations is

partially explained by the absence of sexual dimorphism

with respect to SVL (Valdéz-Gonzalez and Ramirez-

Bautista 2002; Ramirez-Bautista et al. 2013). Walkup et

al. (2017) pointed out that Uta stansburiana, Aspidoscelis

marmorata, A. sexlineata, and Sceloporus consobrinus

present generalist habits in microhabitat choice, being

able to inhabit sites with different degrees of disturbance.

Consequently, these species tend to present reduced

variation in some of their demographic characteristics,

as a measure of phenotypic plasticity towards different

degrees of environmental disturbance.

In this study, lizards from UA were numerically more

abundant and showed a slightly higher survival rate

February 2020 | Volume 14 | Number 1 | e222

Torres Barragan et al.

Table 4. Probability of total survival (@) and recapture (p) for both sexes and season for UA and DA populations. SE = standard

error.

A. Survival values and recapture between sexes

Undisturbed area (UA)

Parameters Estimation SE

® (males) 0.822 0.042

Dp 0.402 0.051

® (females) 0.706 0.056

B. Survival values and recapture between seasons

Undisturbed area (UA)

Parameters Estimation SE

® (dry) 0.765 0.036

® (wet) 0.538 0.074

than those from DA. This suggests that relative density

may be a measure of population variation that predicts

the survival value for both sites. However, in some

cases lower survival probability has been observed in

populations with high density than in populations with

low density (Stearns 1992; Zufiga-Vega et al. 2008).

Survival rates and probabilities of recapture obtained

here for males and females were higher compared to

those reported for Anolis nebulosus (Hernandez-Salinas

2014), Xenosaurus grandis (Zufiiga-Vega et al. 2007), and

Sceloporus grammicus (Pérez-Mendoza et al. 2014). The

models used in these studies considered sex, season (dry and

wet), and populations as variables that express the greatest

sources of variation, similar to the survival rate assessed in

other species of the genus Sceloporus (Zufiga-Vega et al.

2008; Pérez-Mendoza et al. 2014). The results for survival

obtained here were similar to those of other studies (Zufiga-

Vega et al. 2008; Molina-Zuluaga et al. 2013; Hernandez-

Salinas 2014) where survival and recapture rates were

found to be higher in males and females in one population

during the dry season. One possible explanation 1s that the

reproductive activity of this species begins during the rainy

season, causing males to search for mates for reproduction,

participating in male-male agonistic competition, and

consequently becoming more susceptible to predation

(Sinervo et al. 1991). In contrast, for females, survival rates

have been observed to decrease dramatically at the end of

the gestation stage and parturition (end of the rainy season),

possibly because pregnant females become heavier and

slower, therefore increasing the risk of predation during the

wet season (Clobert et al. 1998, 2000; Stearns 1992; Zufiiga-

Vega et al. 2008). This pattern also has been observed in

other lizard species, such as Anolis nebulosus from tropical

dry forest (Ramirez-Bautista and Vitt 1997).

Conclusions

This study found little variation regarding growth rate,

survival, and body size at sexual maturity between

Amphib. Reptile Conserv.

Disturbed area (DA)

Parameters Estimation SE

® (males) 0.75 0.019

Dp 0.40 0.024

® (females) 0.65 0.061

Disturbed area (DA)

Parameters Estimation SE

® (dry) 0.927 0.078

® (wet) 0.489 0.072

populations of Sceloporus spinosus. This supports the

assumption that the toe-clipping method did not impair

the performance of individuals in both populations,

allowing the results of this study to be comparable

with future works. For the above, and contrary to our

expectations, this study showed that males and females

in both UA and DA showed similar growth rate patterns,

with both sexes reaching sexual maturity at similar SVL

in both populations. The results obtained here may not be

regarded as the variation typically observed in these life

history attributes (growth, survival, and SVL at maturity),

since the minimal variation observed in these characters

may indicate that they are genetically fixed components,

as has been determined in some species of Xenosaurus

(Zufiiga-Vega et al. 2005, 2007; Zufiiga-Vega 2011)

and A. nebulosus (Ramirez-Bautista and Vitt 1997).

The results of this study should be taken with caution,

since further studies are needed to determine the effects

of temperature, precipitation, competition, and food on

the life-history characteristics of the species living in

both environments. Additional studies will support more

solid conclusions regarding the growth, survival rate,

and size at sexual maturity across the entire distribution

range. In order to improve these results, we also suggest

the development of hypotheses aimed at determining

the relative abundance of predators. Furthermore, it is

necessary to develop elasticity and sensitivity analyses

with larger sets of capture-recapture data to better

comprehend the effects of environmental pressures on

the life history characteristics of a species that is found in

both intact environments and environments with varying

degrees of disturbance.

Acknowledgments.—We thank Barry Stephenson

for reading and reviewing the English version of this

manuscript, and Graciela Gonzalez Pérez and Gladys

Isabel Manzanero Medina (IPN-CIIDIR-OAXACA)

for academic support. CATB conducted this research as

a student of the Maestria en Ciencias en Conservacion

February 2020 | Volume 14 | Number 1 | e222

Sceloporus spinosus in Oaxaca, Mexico

y Aprovechamiento de Recursos Naturales in the IPN-

CHDIR-OAXACA and with support of a graduate

scholarship from CONACYT (grants 300633). We thank

the National Commission of Natural Protected Areas,

especially Gabriel de Jesus Martinez and Pavel Palacios

Chavez for support and required permits; and Eric N.

Smith (The University of Texas at Arlington) for access to

specimens and information. Special thanks are extended

to Lucio Torres Ruiz and Museum of Vertebrate Zoology

(Berkeley, California, USA), and to Joel Alcantara for

assistance in the field. Finally, we thank the anonymous

referees; your observations and suggestions substantially

improved our work.

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in the lizard Uta stansburiana stejnegeri. American

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comparative population studies in the lizard Uta

stansburiana. Pp. 133-160 In: Systematic

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M.S. Thesis. Instituto Politécnico Nacional, Centro

Interdisciplinario de Investigacion para el Desarrollo

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Carlos A. Torres Barragan is a biologist and herpetologist born in Oaxaca, Mexico. Carlos graduated

from Universidad Autonoma “Benito Juarez” de Oaxaca (UABJO) in Mexico, where he worked on

the ecology of the amphibian and reptile communities of Yagul Natural Protected Area in Oaxaca.

He later earned his Master’s degree at Instituto Politécnico Nacional (IPN-CIIDIR-Oaxaca) in 2015,

where he studied demographic parameters of reptiles living in disturbed and undisturbed sites within

Yagul Natural Protected Area. Currently, Carlos works as a consultant with indigenous communities

in different regions of Oaxaca, in areas regarding biodiversity monitoring, sustainable and friendly-

biodiversity productive systems, and development of local technical capabilities.

Uriel Hernandez Salinas is a Mexican herpetologist interested in the richness, diversity, biogeography,

and conservation of amphibians and reptiles in central and northern Mexico. Uriel is a full-time professor

at CIIDIR Durango of the Instituto Politécnico Nacional and curator of the scientific collection of

amphibians and reptiles at that research institution.

Aurelio Ramirez Bautista is a full-time professor at Universidad Autonoma del Estado de Hidalgo.

Currently, Aurelio teaches undergraduate and graduate biology courses, including those on population

ecology, herpetology, life history evolution, and the reproductive biology of amphibians and reptiles.

His research focuses on the study of biodiversity (species richness), biogeography, and conservation of

amphibians and reptiles of Mexico, as well as life history evolution in lizard species from the temperate,

tropical, and desert environments of Mexico.

February 2020 | Volume 14 | Number 1 | e222

Official journal website:

amphibian-reptile-conservation.org

Amphibian & Reptile Conservation

14(1) [General Section]: 55—62 (e223).

urn:lsid:zoobank.org:pub:A27AAA11-D447-4B3D-8DCD-3F98B13F12AC

A new gecko genus from Zagros Mountains, 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.—A new genus and species of gekkonid lizard is described from the Zagros Mountains, western Iran.

The genus Lakigecko gen.n. can be distinguished from other genera of the Middle East by the combination of

the following characters: depressed tail, strongly flattened head and body, eye ellipsoid (more horizontal), and

approximately circular whorls of tubercles (strongly spinose and keeled).

Keywords. Gekkonidae, Lakigecko gen.n., Lakigecko aaronbaueri sp.n., new species, Reptilia, Sauria, Western Iran

Citation: Torki F. 2020. A new gecko genus from Zagros Mountains, Iran. Amphibian & Reptile Conservation 14(1) [General Section]: 55-62 (e223).

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: 21 October 2018; Accepted: 14 December 2019; Published: 23 February 2020

Introduction

Thus far, 49 species and 14 genera of Gekkonidae have

been recorded in Iran (Safaei-Mahroo et al. 2016). Ten

new species of gekkonid lizards have been described

since 2000, including seven new species and a recently

described monotypic gecko genus, Parsigecko, from

southern Iran (Safaei-Mahroo et al. 2016). This study

adds another new gecko genus to the fauna of Iran

based on the species described herein. In addition, this

study includes a discussion of the validity of genus

Carinatogecko Golubev and Szczerbak, 1981.

Materials and Methods

During field trips carried out on the Iranian plateau,

the new taxon was collected from Nourabad, Lorestan

Province, Zagros Mountain, western Iran (Fig. 1).

The holotype and paratype specimens of this new

species have been deposited in the MTD (Senckenberg

Naturhistorische Sammlungen, Museum fir Tierkunde,

Dresden, Germany).

Measurements. The following measurements were taken

with a digital caliper, and interpolated to the nearest

0.1 mm: SVL: snout-vent length (from tip of snout

to anterior margin of cloaca); TaL: tail length (from

posterior margin of cloaca to tip of tail, original or

regenerated); TaL/TL: ratio tail length/total length; TW:

tail width (taken at base of tail); TH: tail height (taken

at base of tail); TW/TH: ratio tail width/tail height; HL:

Correspondence. torkifarhang@yahoo.com

Amphib. Reptile Conserv.

head length (from tip of snout to posterior margin of ear);

HH: maximum head height (from occiput to undersides

of jaws); HW: maximum head width; HL/SVL: ratio

head length/snout-vent length; EDH: eye diameter

horizontal; EDV: eye diameter vertical; IO1: anterior

interorbital distance; IO2: posterior interorbital distance;

EE: distance between posterior margin of eye to posterior

margin of ear opening; EaD: ear diameter vertically; IN:

internostril distance; IL: interlimbs; FL: forelimb length;

and HL: hindlimb length.

Lepidosis. The following scale counts were taken, and

on the right side of body where appropriate: N: nasals

(nasorostrals, supranasals, postnasals, first supralabial);

ISN: intersupranasals (scales between supranasals, in

contact with rostral); SPL: supralabials; IFL: infralabials;

IO: interorbitals (number of scales in a line between

anterior corners of eyes); PO: preorbitals (number of

scales in a line from nostril to anterior corner of the eye);

PM: postmentals; LT: number of subdigital lamellae (on

toes counted from the first broad lamella to the claw);

DC: number of dorsal crossbars; and TC: number of tail

crossbars.

Comparisons. For comparison, representatives of five

other naked-toed gekkonid genera were examined and

compared with the new genus as follows: Cyrtopodion

(C. scabrum: n = 9, FTHM 004600-08:; C. kiabii: n =

8, FTHM 004880-87); Tenuidactylus (T: caspius: n =

11, FTHM 004851-61); Mediodactylus (M. russowii,

n= 2; M. heterocercus, n = 5; M. sagittifer, n = 3; M.

February 2020 | Volume 14 | Number 1 | e223

A new gecko genus from Zagros Mountains, Iran

Type Locality

____— Lorestan Province

Iranian Plateau .

Fig. 1. Map showing the type locality of Lakigecko aaronbaueri

sp.n. in the Garmabe region, Nourabad, Lorestan Province,

Tran.

spinicauda, n = 3), Carinatogecko (C. heteropholis: n=

29; FTHM 003400-003428; C. aspratilis, n= 12, FTHM

003300-003311; C. ilamensis, n = 3), Bunopus (B.

tuberculatus: n= 12, FTHM 003200-11). Other data were

taken from Minton et al. (1970), Anderson (1973, 1999),

Golubev and Szczerbak (1981), Nazarov et al. (2009),

Ahmadzadeh et al. (2011), Fathinia et al. (2011), Torki

(2011), and Safaei-Mahroo et al. (2016). Abbreviations:

TT: Tail tubercle distribution (D: dorsal; L: dorsolaterals;

V: ventral); TS: tail shape (S: squat, 1.e., short and thick:

Q: not squat, i.e., more elongate); LT: Lamella type (M:

smooth; K: keeled); DT: dorsal tubercles in each segment

(C: in contact; D: not in contact); DS: dorsal scales

between whorls (E: elevated; N: not elevated); GS: digits

shape (A: angular; S: sub-angular; W: weakly angular;

N: not angular); FP: femoral pores (+: present; -: absent);

ES: eye shape (G: Globular; E: Ellipsoid); and HA: head

angle (degree of lateral view between lower ear-nostril-

upper eye).

Systematics

Lakigecko gen.n.

urn:lsid:zoobank.org:act:0FA39E43-910A-4F0C-902A-48871 4031507

Type species: Lakigecko aaronbaueri sp.n., herein

described.

Definition: The monotypic genus Lakigecko gen.n.

differs from all other genera of the Family Gekkonidae

by the following combination of characters: (1)

depressed tail; (2) much flattened head and body; (3)

acute head angle (laterally); (4) eye horizontally ellipsoid

(horizontal/vertical ratio in life more than 1.2 and

preserved approximately 1.6); (5) whorls of tubercles

Amphib. Reptile Conserv.

Table 1. Morphometric measurements (in mm) and scalation

of Lakigecko aaronbaueri sp.n., abbreviations as defined in the

text. * end of tail regenerated; ** most part of tail regenerated.

Holotype Paratype

Charatters (MTD 49500) (MTD 49499)

SVL 32.79 32.19

TaL 25.58* 25.75%

TaL/TL 0.78 0.80

TW 3.06 3.24

TH 2.48 1.95

TW/TH 1.23 1.66

HL 8.04 8.10

HH 3.48 3.40

HW 6.45 6.27

HL/SVL 0.24 0.25

EDH 2.95 2.30

EDV 121 1.59

101 3.27 3.23

102 4.38 4.58

EE 2.68 2.78

EaD 0.70 0.63

IN 0.98 1.02

IL 14.80 13.32

FLL 11.81 11.26

HLL 16.90 15.91

N 5 5

ISN 1 1

SPL 9 10

IFL 7 7

IOS 13 12

PO 9 10

PM 3 3

LAT 17 19

approximately circular (extended onto ventrolateral

surface): strongly spinose and keeled caudal tubercles

that extend onto dorsolateral and ventrolateral surfaces,

lateral tubercles larger than mid-dorsal and ventral, mid-

dorsal and ventral tubercles are clearly different from

smaller subcaudal scales, whorls separated by two rows

of keeled scales; (6) spinose tubercles extend along entire

length of tail (tubercle shape consistent throughout); (7)

mid-dorsal tubercle rows separated from each other by

two scales, dorsolateral rows of tubercles (four tubercle

rows on each side) are in contact with one another; (8)

dorsolateral scales strongly elevated, pyramidal, and

keeled; (9) dorsal scales mostly smooth; and (10) without

femoral or precloacal pores; two postcloacal pores

(openings to the postcloacal sacs).

Distribution: Known only from the type locality.

Etymology: The generic nomen Lakigecko is derived

from the word “Laki” which refers to the Lak region near

the type locality.

Comparisons (Fig. 2, Table 2): Differs from

Cyrtopodion Fitzinger, 1843 by having small, spiny,

and strongly keeled subcaudal scales (vs. smooth, plate-

February 2020 | Volume 14 | Number 1 | e223

Fig. 2. Comparisons of (1) head shape, (2) dorsal view, (3) dorsal view of tail, (4) lateral view of tail, (5) ventral view of tail, and

(6) subdigital lamella of hindlimbs, in the new gecko with other genera. (a) Bunopus tuberculatus, (b) Tenuidactylus caspius, (Cc)

Cyrtopodion scabrum, (da) Carinatogecko heteropholis, (e) Mediodactylus russowii, and (f) Lakigecko aaronbaueri sp.n. Photos by

F- Torki (a-d, f) and B. Safaei-Mahroo (e).

like subcaudals); dorsal scales elevated and overlapping (vs. not). In general, the new genus is easily differentiated

(vs. non-elevated or flattened cobble-stone shaped) anda = from = Cyrtopodion, Tenuidactylus, Mediodactylus, and

greatly flattened and elongate head (vs. head oval and more —_ Carinatogecko by its acute head angle (less than 30° vs.

massive); from Mediodactylus Szczerbak and Golubev, 40°, 40°, 35°, and 35°, respectively), strongly flattened

1977 by having keeled, tubercular scales forming the end _ head, depressed tail, more caudal tubercles, spinose caudal

of each caudal segment (vs. in the middle of each segment), tubercles, and elliptical eye shape.

at least 14 sharply keeled tubercles in each caudal whorl Lakigecko gen.n. can be distinguished from Agamura

(vs. a semicircle of six whorls: three on left and three on —_ Blanford, 1874 by its large and strongly keeled caudal

right), caudal tubercles in contact with one another laterally — tubercles (vs. not), limbs robust (vs. slender), enlarged

(vs. not in contact), caudal tubercles forming a relatively — postmentals (vs. not enlarged), and tail broad (vs. slender);

complete ring around the tail (vs. distributed only on the from Bunopus Blanford, 1874, by subdigital lamellae

dorsal half of the tail), and a flattened and elongate head(vs. | completely smooth (vs. subdigital lamellae with free

not); from Carinatogecko Golubev and Szczerbak, 1981 by = margin under magnification); 14 sharply keeled tubercles

smooth subdigital lamellae on the forelimbs and hindlimbs —_in each whorl segment of tail (vs. 4-6 and not sharp and

(vs. keeled transverse subdigital lamellae), and smooth spiny); from Rhinogecko de Witte, 1973 by nasal scales

dorsal scales (vs. strongly keeled); from Yenuidactylus not forming a cylindrical caruncle (vs. nostril at apex of

Szczerbak and Golubev, 1984 by the lack of femoral prominent swollen or cylindrical caruncle formed by the

pores (vs. more than 20 femoral pores), ventral subcaudals _ nasal scales); from Microgecko and Tropiocolotes Peters,

small and strongly keeled (vs. plate-like and smooth), 14 ~—=1880 by having uniform dorsal scales, large scales, and

sharply keeled tubercles in each tail whorl segment (vs. six _ keeled tubercles on the dorsum of body and tail (vs. dorsal

tubercles per whorl); and a flattened and elongated head __ scales heterogeneous, scales small, and without keeled

Table 2. Comparisons of Lakigecko gen.n. with other related genera. For abbreviations see Materials and Methods section.

Characters ar ns nn a TS LT DT DS GS FP ES HA

Cyrtopodion + +- - B M CG N A - G >40°

Tenuidactylus + +- - B M GC N A + G >40°

Bunopus + ah - B K C N N - G >40°

Mediodactylus + lh ~ B M D N S - G ca

Carinatogecko =r: +- - B K D E W - G 235°

Lakigecko gen.n. sty + ats S M @ E S - EB <30°

Amphib. Reptile Conserv. 57 February 2020 | Volume 14 | Number 1 | e223

A new gecko genus from Zagros Mountains, Iran

Fig. 3. (a) Dorsal, (b) ventral, (c) lateral head, and (d) lateral

tail views of the holotype of Lakigecko aaronbaueri sp.n. Red

stars: tubercular whorls; white star: keeled scale interspaces.

tubercles on dorsum and tail); from Hemidactylus by

non-dilated digits (vs. well-defined dilated digital bases);

from Pseudoceramodactylus Haas, 1957, Crossobamon

Boettger, 1888, and Stenodactylus Fitzinger, 1826 and

Teratoscincus by digits without elongate fringes (vs.

digits with well-defined lateral elongate fringes); from

Parsigecko Safaei-Mahroo, Ghaffari, and Anderson,

2016 by large, keeled tubercles on body dorsum,

subcaudal scales small and strongly keeled (vs. without

dorsal tubercles, and subcaudal scales large and smooth).

Contents: At present the new genus includes a single

species, Lakigecko aaronbaueri sp.n.

Lakigecko aaronbaueri sp.n. (Fig. 3-4)

Amphib. Reptile Conserv.

Fig. 4. Dorsal view in life of (a) holotype and (b) paratype of

Lakigecko aaronbaueri sp.n. before preservation.

urn:Isid:zoobank.org:act:9809A55C-652A-4510-B39A-753EA57CC81D

Holotype (Fig. 3): MTD 49500, adult male, collected

on the western slope of the central Zagros Mountains,

Garmabe Region, Nourabad, Lorestan Province, western

Iran on 8 April 2016 at night (34°03’N, 47°28’E,

elevation 1,470 m asl) by Farhang Torki.

Paratype (Fig. 4): Adult male, MTD 49499, same data

as for holotype.

Description of holotype: Body flattened; head flattened;

tail depressed and strongly covered by sharp tubercles,

eye opening clear, pupil vertical.

Dorsal body. Nostril surrounded by five scales (rostral,

nasal, and three behind); nasal nearly completely divided;

large dorsal tubercles strongly keeled, each dorsal tubercle

surrounded by 10 (or 11) scales, 13 dorsal tubercle rows

on dorsal, mid-dorsal tubercles separated from each other

by one or two scales, dorsolateral tubercles (four on each

side of vertebral midline) are in contact with one another,

eight rows (4+4: left + right) extend onto tail, dorsolateral

tubercles mostly in contact with one another, separated

by two small scales in the dorsal midline; most dorsal

scales small and smooth, a few scales large and keeled,

mid-dorsal scales elevated, dorsolateral scales strongly

elevated, keeled and pyramidal in form; keeled dorsal

February 2020 | Volume 14 | Number 1 | e223

Torki

3 es

Fs. » a

_ a .

2 tg Vid od Fs ye) if:

tubercles extend onto neck and occipital region; neck

tubercles pyramidal in form, but dorsal tubercles less

so; middorsal neck scales swollen (similar to granules)

and not overlapping or elevated, dorsolateral neck scales

strongly swollen and keeled (pyramidal) and extending

in front of and behind ear; neck scales are different in

shape compared to dorsum; upper head tubercles (keeled)

smaller than dorsum; nearly all head scales granule-like,

keeled and heterogeneous (small to large, cycloid to

polygonal); interorbital scales keeled and heterogeneous

in size and shape; snout scales keeled; arm covered

by strongly keeled large scales (same size as dorsal

tubercles); elbow scales keeled; tubercles and scales

on forearm strongly keeled; digits covered by strongly

keeled scales; leg covered by large, strongly keeled,

and overlapping scales (similar size to dorsal tubercles),

keeled scales only distributed near knee, lateral surface

of leg covered by small scales; foreleg with strongly

keeled tubercles distributed among keeled scales, back

of hand and digits with strongly keeled scales.

Ventral body. Mental pentagonal; supralabials nine;

infralabials seven; three postmentals (PM), 1‘' PMs in

contact with one another, 2™ PMs separated by two scales,

3 PMs separated by six scales, PMs surrounded by 12

scales, 1‘ PM in contact with 1% infralabials, 2"? PMs

Amphib. Reptile Conserv.

i =

Fig. 5. Habitat of Lakigecko aaronbaueri sp.n. at the type locality in the Garmabe region, Nourabad, Lorestan Province, Iran.

in contact with 2™ infralabials, and 3™ PMs in contact

with 2™ and 3" infralabials; chin scales small, round, and

smooth; scales on neck and between forelimbs smooth and

overlapping; interlimb scales keeled, and less overlapped,

scale rugosity increases from proximal (across forelimbs)

to distal (across hindlimbs) of interlimbs; scales between

hindlimb bases as well as preanal scales large and smooth

(not keeled); arm scales keeled, elevated; forearm scales

keeled and mostly trihedral; forearm scales larger and

more rugose than upper arm; scales on palm granular;

lamella swollen; dorsolateral scales on digits sharp; thigh

as well as foreleg scales keeled and elevated, foreleg

scales more rugose than those on thigh; small scales on

lateral surface of leg; scales on soles granule-like with

some very similar to lamellae, distal lamella smaller;

lamellae swollen; dorsolateral scales on digits sharp,

lamellae on the middle of each finger (at joint) larger than

others; number of lamellae under toes 10:14:15:17:16.

Tail. Mostly original, end of tail regenerated; eight rows

of dorsal tubercles extended onto tail; tail tubercles

strongly keeled, elevated, sharply keeled, and in

contact with each other within each whorl; each whorl

formed by 18 (2+8+6+2) tubercles as follows: two in

the middle and different from pyramidal scales, eight

(4+4) large and elevated and sharp tubercles on dorsum

February 2020 | Volume 14 | Number 1 | e223

A new gecko genus from Zagros Mountains, Iran

and extend ventrolaterally, six small (3+3) ventrolateral

tubercles distinguishable from subcaudal scales, and

finally two small scales that are similar to subcaudal

scales; 14 whorls of tubercles are clear on tail, each

separated from one another dorsally by two small scales

and ventrally by two keeled scales; two rows of small

and strongly keeled scales between each whorl (from 1*

to 12" whorls), three such rows of scales between 13"

and 14" circles.

Color pattern. Background of dorsal body brown, with

dark bars covering dorsum and tail; dorsal bars irregular

and reticulating; nine dark bars on tail, interspaces

yellowish; ventral dirty white; middle of subcaudals

bright grayish, lateral surfaces dark grayish; four

irregular longitudinal bars on occipital region; dorsum

of head covered by small spots; width of bars (wider

than interspaces) covering forelimbs (seven bars) and

hindlimbs (seven bars); limb bars extend onto digits,

five bars on longer digits and four bars on others; dorsal

scales on body, tail, limbs, and head are pigmented;

ventral body scales pigmented, pigmentation more

pronounced laterally than midventrally, two rows

of small precloacal scales exhibit the maximum

pigmentation, large precloacal scales have minimum

pigmentation; scales on venter of leg and foreleg

pigmented; all lamellae of pes are dark brownish, middle

lamellae less dark; lamellae on digits of manus are

light brown, scales on palm are brownish or ashy; chin

scales less pigmented; mental, labials, and postmentals

pigmented. Color pattern in preservative is similar to

life, but paler.

Description of Paratype

Most characters are similar to holotype, some differences

are shown in Table 1 and others are: 13 dorsal tubercle

rows, four lateral tubercles are in contact with each other

and middle tubercles are separated by small scales (same

as holotype); tail regenerated, all dorsal and ventral scales

are small, strongly keeled, and similar to one another,

dorsal scales regular and ventral scales less regular,

strongly elevated and overlapping; number of lamellae

under toes 11:13:18:19:15.

Color pattern. Five dark irregular bars between limbs;

one V-shaped bar on neck; one arcuate bar on occipital;

head covered by small spots; all dorsal body bars

and spots are blackish; background of dorsal body

grayish; tubercles on bars are black and most of those

in interspaces are brownish (some of them are both

black and brownish in color); regular and irregular bars

covering limbs and digits; chin white; ventrals of body

and limbs are dirty white; irregular darkish stripes cover

dorsum of regenerated tail, all dorsal and ventral scales

pigmented; color pattern in preservative is similar to life,

but colors are more pale.

Amphib. Reptile Conserv.

Etymology. The species name “aaronbaueri’ refers

to Professor Aaron M. Bauer (Villanova University,

Villanova, Pennsylvania, USA) for his major contributions

to works on the systematics and morphology of the

geckos of the world.

Habitats and ecology. The specimens of L. aaronbaueri

sp.n. were found between stones in the Garmabe region

(Nourabad, Lorestan Province, Iran). The type locality

is characterized by oak forested hills surrounded by

mountains, and is adjacent to the Giz-e-roo River (Fig. 5).

Specimens were obtained from the top of the mountains.

The new species is syntopic with three reptiles, Laudakia

nupta, Ophisops elegans, and Heremites aurata.

Distribution. Lakigecko aaronbaueri sp.n. is at present

known only from the type locality.

Discussion

Previously Cervenka et al. (2010) suggested that

Carinatogecko belonged in the synonymy of

Mediodactylus. Based on my collections from the

Shorab region near Khorramabad (the locality of

Carinatogecko cf. heteropholis, REPT/IRA/1139),

Cervenka et al. (2010) worked on Mediodactylus sp.

from Lorestan Province, not Carinatogecko. This

mistake occurred due to an erroneous taxonomic

diagnosis based on two criteria. First, the dorsal scalation

type and shape (Cervenka et al. 2010; Torki 2011) of

Carinatogecko cf. heteropholis (REPT/IRA/1139) does

not support this species belonging to Carinatogecko,

as the author has previously discussed (Torki 2011).

Second, the subdigital lamellae of the new collection

do not match the condition of Carinatogecko (smooth

vs. keeled). Therefore, I suggest that Carinatogecko

is valid and distinct from the other genera of the

Mediodactylus-Cyrtopodion-Tenuidactylus group as

well as from Lakigecko gen.n. The number of lamellae,

a character cited by Cervenka et al. (2010) to confirm

that their specimens belonged to Mediodactylus,

is not an important taxonomic character for the

distinguishing of Carinatogecko from Mediodactylus.

Also, the typical cryptic color patterns on the dorsum

and tail in Mediodactylus are not M- or V-shaped dark

transverse bars, as reported by Cervenka et al. (2010).

Based on my personal observations, there is more

variability in the dorsal color patterns in Mediodactylus

sp. and Carinatogecko sp., such as zigzag forms and

completely irregular dorsal patterns (this variability is

clear in urban house geckos). On the other hand, based

on my observations, Mediodactylus sp. are distributed

mostly in urban houses in Iran, as a result of human

activity, rather than in the surrounding habitats. This is

also true for some other geckos, such as Cyrtopodion

and Hemidactylus, and for some hair-like pad geckos

(Torki 2007; Torki et al. 2008). Carinatogecko may

February 2020 | Volume 14 | Number 1 | e223

Torki

be defined on the basis of at least three important

taxonomic characters: (1) all body scales keeled (except

intermaxillaries, nasals, chin shields, and_labials),

(2) subdigital lamellae keeled (not smooth), and (3)

subcaudal scales small and strongly keeled. In any case,

gecko taxonomy in the Zagros Mountains 1s a complex

problem, largely due to incomplete and poor taxonomic

studies to date.

Tail tubercles in Cyrtopodion, Tenuidactylus, Bunopus,

and Lakigecko gen.n. are in contact with one another

within whorls, and this is in contrast to Mediodactylus and

Carinatogecko (where they are separated). This character

is inconsistent with a recent phylogenetic study (Bauer et

al. 2013) that includes some of these taxa. 7enuidactylus,

Bunopus, and Cyrtopodion are more closely related to

each other and collectively have a sister-taxon relationship

with Mediodactylus. Both the highly keeled and ventrally

extending tubercles in Lakigecko gen.n. and the shape

of the tail differ from other genera (7enuidactylus,

Bunopus, Cyrtopodion, Mediodactylus, Carinatogecko).

Additionally, the eye shape (horizontally elliptical) and

head shape (greatly flattened) of Lakigecko gen.n. are

different from those in these genera. Although, I have not

studied the skull anatomy of Lakigecko gen.n. directly,

the external morphology of the head suggests a distinctive

skull anatomy of the new genus as well.

In general, four characters of Lakigecko gen.n. are

completely different from related genera (7enuidactylus,

Bunopus, Cyrtopodion, Mediodactylus, and Carinato-

gecko) including (Table 2): (1) tail tuberculation, (2) tail

shape, (3) eye shape, and (4) head shape. On this basis I

suggest that Lakigecko gen.n. is likely sister to the group

Tenuidactylus-Bunopus-Cyrtopodion + Mediodactylus-

Carinatogecko.

Key to the genera of Gekkonidae in Iran (after Anderson 1999; Safaei-Mahroo et al. 2016)

la“ Bieits-stronely dilated + 8s PROS c-2st Roy E de

Hb? Toieitsmotdilated, 0%. i SRE oR nie Al eA cele

2a: Digits with well-defined lateral fringe of elongate scales

2b: Digits without fringe of elongate scales.........................

3a: Small dorsal scales intermixed with rounded tubercles....

3b: Dorsal scales uniform, not intermixed with tubercles......

4a: Enlarged postmental scales present.......................00.0..

4b: No enlarged postmentals..........0..00 0.0000 c cece cece ee cece

5a: Dorsal scales heterogeneous...............0.0 000.0. .c0 cee cece

5b: Dorsal scales small (granule like), homogeneous..........

phat Brien de Serer Pease ee ehh ce Pseudoceramodactylus Haas, 1957

opts, se dee ak rode rt she Theme ie eee a of te Stenodactylus Fitzinger, 1826

6a: Nostril at apex of prominent swollen or cylindrical caruncle formed by nasal scales, rostral excluded from border of

MOSH. tate, tee cere ae Reeth eet tees ha tet atane eee acta eee enlace ioe ict chars ted ae hee ee Rhinogecko de Witte, 1973

6b: Nasal scales do not form cylindrical caruncle, although they may appear to be swollen around the nostril; rostral normally

foriissparl:ol-bordereOf MOS le = xt eeke sco seers are ery ony hvumy Ohare de. pearson eae ok emanate WR hey play em 5 te mereen: Senate 7

7a: Tail cylindrical, very slender, and of almost uniform diameter from base to tip (tip blunt), no mucronate tubercles or

UATTULL RS, eee a wn BOS PL OE EERE NT Ie eek Rue Seam SL OF Faas ey ey aS Ree ee Agamura Blanford, 1874

8b: Tail depressed, tail tubercles strongly keeled, spinose, in contact together in at least 14 rows in each whorl, whorl tubercles

extended in laterals and ventral of tail; head angle (laterally) less than 30°............... 0.0... 0c cece eee eeees Lakigecko gen.n.

Sbeclail different’ trom: above, shead-angle™(lateralls> more: that 33° ie...) hs. e564 5: 8s Sok A ew noe old BE, wba tee »

Oa SUlbdiGiial ylarneliac. Keele: se). ha: Bika: ba ete bss bi ols whe ole rures 5. df Se carmomy' dern§ 5 sion a3 8 ore eRe By, Ral ok Re be PI OR ad a 10

Di Subaraital lanmellacr SMGOt de eta. Ake ec eee ye Ae ee mee ok oe RN nc ees inte Re Na scaly et ey epee tal. Ae, ae ee 1]

10a: All scales on body except chin shields, supralabials, and infralabials keeled; dorsal tubercles in each caudal segment not in

contact: subcaudalscales. not*denticulate: «2.050. 520.2 e.a. crib Gee. Aes Carinatogecko Golubev and Szczerbak, 1981

10b: All body scales are not keeled; dorsal tubercles in each segment are in contact; subcaudal scales denticu-

TAGS stip chats soe eee theese Lela Oh I ig RUA AUER, Mae, hse ts AE a RR eM MUTI 8 COE wy AP EO SE. Bunopus Blanford, 1874

lla: Digit shape sub-angular, spine-shaped caudal tubercles do not contact each other in the whorls of a

SCOMIENIN Woe AM ek Ua eh eee ee, ey renee Moe, ewe 48 De Mediodactylus Szczerbak and Golubev, 1977

1lb: Digit shape strongly angular, low, or moderately high tail tubercles broadly contact each other across the dorsum

of tail whorls and are surrounded by one to two smaller scales..........00.0...0 cocoon cece ccc ence cette cece eeee ees b2

as Femoral: pores presente 8s 1. cee ce hao denet, elma ila aude ewe hss tree ae At Tenuidactylus Szczerbak and Golubev, 1984

Pbk emaGral S9OTCS AWSOME, ex, Sein soretan ds o-5 anyone ate che ceang Macaig thie Beal emne nd, 20.5 ay ph Pad sees Lae Bu Cyrtopodion Fitzinger, 1843

13a: Subcaudals arranged in single large, broad row, two strongly keeled tubercles on either side of each

ITVS eros sat es. alot a cee oes Teese pa eae ce he a Maint oat senhuge/nc fs Papen Rasta Parsigecko Safaei-Mahroo, Ghaffari, and Anderson, 2016

Ibs Varlkscales unirorn., hOmMeeenOUS:, NOE KEGICd! ...15.My. tpi, SAT! Sons Bhe Berle tne each Sic Rin Bee der Ml eth ery hue eB UR Briel rets LBM 2! 14

jE ne Ae yilaye ikea Les Hp ba Na tied UFeYotal @ ete (er6 | out tm es oh OO Ae A tat, A eA RE cr Re eat Pht re Tropiocolotes Peters, 1880

14b--Subdisitallamel lac smeothy: 2.5 seta turtun tens op bent clad aceigeney nah uses stun Sebo eree nant eee ua oe beeen Microgecko Nikolsky, 1907

Amphib. Reptile Conserv. 61 February 2020 | Volume 14 | Number 1 | e223

A new gecko genus from Zagros Mountains, Iran

Acknowledgments

I wish to thank Professor Aaron M. Bauer (Villanova

University, Villanova, Pennsylvania, USA), Craig

Hassapakis (Utah, USA), and Michael L. Grieneisen

(California, USA) for editing and improving my

manuscript. I wish to thank Barbod Safaei-Mahroo (Pars

Herpetologists Institute, Tehran, Iran) and Nasrin Heidar1

(Divandare, Kurdistan, Iran) for preparing photographs

of Mediodactylus and Carinatogecko ilamensis.

Literature Cited

Ahmadzadeh F, Flecks M, Torki F, Bohme W. 2011. A

new species of angular-toed gecko, genus Cyrtopodion

(Squamata: Gekkonidae), from southern Iran. Zootaxa

2924: 22-32.

Anderson SC. 1973. A new species of Bunopus (Reptilia:

Gekkonidae) from Iran and a key to lizards of the

genus Bunopus. Herpetologica 29: 355-358.

Anderson SC. 1999. The Lizards of Iran. Society for the

Study of Amphibians and Reptiles, Ithaca, New York,

USA. 442 p.

Bauer AM, Masroor R, Titus-Mcquillan J, Heinicke

MP, Daza JD, Jackman TR. 2013. A preliminary

phylogeny of the Palearctic naked-toed geckos

(Reptilia: Squamata: Gekkonidae) with taxonomic

implications. Zootaxa 3599(4): 301-324.

Cervenka J, Kratochvil L, Frynta D. 2010. Phylogenetic

relationships of the gecko genus Carinatogecko

(Reptilia: Gekkonidae). Zootaxa 2636: 59-64.

Fathinia B, Karamiani R, Darvishnia H, Heidari

N, Rastegar-Pouyani N. 2011. A new species of

Carinatogecko (Sauria: Gekkonidae) from Ilam

Province, western Iran. Amphibian & Reptile

Conservation 5(1): 61—74 (e33).

Golubev ML, Szczerbak NN. 1981. Carinatogecko gen.n.

(Reptilia, Gekkonidae) — Novy} rod gekkonovykh

jashcheric iz jugo-zapadnoj Asi. Vestnik Zoologii

1981(5): 34-41.

Minton SA, Anderson SC, Anderson JA. 1970. Remarks

on some geckos from southwest Asia, with description

of three new forms and key to the genus Tropiocolotes.

Proceedings of the California Academy of Sciences

37(9): 333-362.

Nazarov R, Ananjeva N, Radjabizadeh M. 2009. Two

new species of angular-toed geckoes (Squamata:

Gekkonidae) from south Iran. Russian Journal of

Herpetology 16: 311-324.

Safaei-Mahroo B, Ghaffari H, Anderson SC. 2016. A

new genus and species of gekkonid lizard (Squamata:

Gekkota: Gekkonidae) from Hormozgan Province

with a revised key to gekkonid genera of Iran. Zootaxa

4109(4): 428-444.

Torki F. 2007. A note of some ecological and social

aspects of geckos in Iran. Chit-Chat 19: 8-11.

Torki F. 2011. Description of a new species of

Carinatogecko (Squamata: Gekkonidae) from Iran.

Salamandra 47: 103-111.

Torki F, Gharzi A, Nazari-Serenjeh F, Javanmardi S,

Heidari N. 2008. Geckos of the genera Tropiocolotes

and Asaccus in the Zagros Mountains, Iran. Gekko

5(2): 31-43.

Torki F, Ahmadzadeh F, Ilgaz C, Avci A, Kumlutas Y.

2011. Description of four new Asaccus Dixon and

Anderson, 1973 (Reptilia: Phyllodactylidae) from

Iran and Turkey. Amphibia-Reptilia 32: 185-202.

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 herpetofauna 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) in 2018. Currently,

Farhang is studying the evolution and developmental biology, based on mathematical methods.

Iran University of Medical Sciences supported his research during 2018.

Amphib. Reptile Conserv.

February 2020 | Volume 14 | Number 1 | e223

Se * ~ ne

Chiropterotriton chico Garcia-Castillo, Rovito, Wake, and Parra-Olea 2017. El Chico Salamander is a state endemic

species known only from Parque Nacional El Chico, at elevations from 2,400 to 3,050 m in pine-oak forest (Frost

2019). In the original description the authors noted that “it is unlikely to occur more widely, because surrounding areas

have been extensively surveyed” (Garcia-Castillo et al. 2017: 502). Also noted in the original description is that “this

species was previously considered as conspecific with C. multidentatus and occurs in sympatry with C. dimidiatus

and Aquiloeurycea cephalica. Likewise, [sthmura bellii has been collected very near sites where C. chico was once

common..., but it is unknown if the two species occur in syntopy” (Garcia-Castillo et al. 2017: 502). Garcia-Castillo et

al. (2017: 502—503) noted that this salamander was “incredibly abundant” when one of the authors of this paper (David

B. Wake) visited the park in August of 1971, but that by the mid-1970s it had become “uncommon and then rare.”

These authors also indicated (p. 503) that the decline of C. chico apparently is not related to habitat loss or disturbance.

They concluded that this salamander should be judged to be Critically Endangered, based on the IUCN criteria B1

ab(v). Finally, Garcia-Castillo et al. (2017: 503) noted that “Hidalgo includes an unusual region where two of the main

mountain complexes of Mexico meet: the Trans-Mexican Volcanic Belt (TMVB) and Sierra Madre Oriental (SMO).

These ranges are known to have a high degree of topographic, geologic and climatic variability [that] has promoted

a high biodiversity... This is especially true for the herpetofauna from Hidalgo...which is the state with the highest

number of species of Chiropterotriton (6), representing 37.5% of the described species of this genus.” Photo by Sean

Rovito, courtesy of Mirna G. Garcta-Castillo.

Official journal website:

amphibian-reptile-conservation.org

Amphibian & Reptile Conservation

14(1) [General Section]: 63-118 (e224).

The herpetofauna of Hidalgo, Mexico: composition,

distribution, and conservation status

‘Aurelio Ramirez-Bautista, 7Uriel Hernandez-Salinas, ‘Raciel Cruz-Elizalde,

3Christian Berriozabal-Islas, ‘Israel Moreno-Lara, ‘Dominic L. DeSantis, *Jerry D. Johnson,

SEli Garcia-Padilla, °Vicente Mata-Silva, and ’*Larry David Wilson

‘Laboratorio de Ecologia de Poblaciones, Centro de Investigaciones Bioldgicas, Instituto de Ciencias Basicas e Ingenieria, Universidad Aut6noma

del Estado de Hidalgo, Km 4.5 Carretera Pachuca-Tulancingo, 42184 Mineral de La Reforma, Hidalgo, MEXICO ?Instituto Politécnico Nacional,

CIIDIR Unidad Durango, Sigma 119, Fraccionamiento 20 de Noviembre II, Durango 34220, MEXICO *Programa Educativo de Ingenieria en

Biotecnologia. Universidad Politécnica de Quintana Roo. Av. Arco Bicentenario, M 11, Lote 1119-33, Sm 255, 77500 Cancun, Quintana Roo.

MEXICO *Department of Biological and Environmental Sciences, Georgia College and State University, Milledgeville, Georgia 31061, USA

‘Department of Biological Sciences, The University of Texas at El Paso, El Paso, Texas 79968-0500, USA °Oaxaca de Judrez, Oaxaca 68023,

MEXICO ‘Centro Zamorano de Biodiversidad, Escuela Agricola Panamericana Zamorano, Departamento de Francisco Morazan, HONDURAS

817350 Pelican Court, Homestead, Florida 33035-1031, USA

Abstract.—The herpetofauna of Hidalgo, Mexico, is comprised of 203 species, including 42 anurans, 17

caudates, one crocodylian, 137 squamates, and six turtles. Here, the distribution of the herpetofaunal species

are catalogued among the four recognized physiographic regions. The total number of species varies from

77 in the Mexican Plateau to 166 in the Sierra Madre Oriental. The individual species occupy from one to four

regions (mean = 2.1). About 69% of the Hidalgo herpetofauna is found in only one or two of the four regions,

which is of considerable conservation significance. The greatest number of single-region species occupies the

Sierra Madre Oriental (25), followed by the Gulf Coastal Lowlands (15), the Trans-Mexican Volcanic Belt (6), and

the Mexican Plateau (2). The Coefficient of Biogeographic Resemblance (CBR) indicates that the Sierra Madre

Oriental and the Gulf Coastal lowlands share the most species (72), because of their adjacent geographic

position and they contain a significant number of generalist species that occur in the Gulf lowlands of Mexico,

southern USA, Central America, and/or South America. The two largest geographic regions in Hidalgo by area,

Sierra Madre Oriental and Mexican Plateau, reflect opposite patterns in species richness (166 and 77 species,

respectively) due to overall differences in the ecological characteristics between them. A similarity dendrogram

based on the Unweighted Pair Group Method with Arithmetic Averages (UPGMA) depicts two distinct clusters,

one between the Sierra Madre Oriental and Gulf Coastal Lowlands and the other between the Mexican Plateau

and Trans-Mexican Volcanic Belt. The former cluster reflects the two regions sharing a substantial number

of herpetofaunal species that occur on the Gulf lowlands of North America and Central America, as well as

a few that enter South America. The second cluster is due to the two montane regions being adjacent to one

another and their ecological similarities. With respect to the distributional categories, the largest number of

species is that of the country endemics (104 of 203), followed by non-endemics (92), state endemics (four),

and non-natives (three). The principal environmental threats to the Hidalgo herpetofauna are deforestation,

livestock, roads, pollution of water sources, cultural factors, and diseases. The conservation status of each

native species was assessed by means of the SEMARNAT (NOM-059), IUCN, and EVS systems, of which the

EVS system was the most useful. The Relative Herpetofaunal Priority (RHP) method was also used to designate

the rank order significance of the physiographic regions and the highest values were found for the Sierra

Madre Oriental. Most of the five protected areas in Hidalgo are located in the Trans-Mexican Volcanic Belt,

which is only the second most important region from a conservation perspective. In addition, only 78 of the 200

native species found in Hidalgo are recorded in total from the five protected areas. Finally, a set of conclusions

and recommendations are offered for the future protection of the Hidalgo herpetofauna.

Keywords. Anurans, caudates, crocodylians, physiographic regions, protected areas, protection recommendations,

squamates, turtles

Resumen.—La herpetofauna de Hidalgo, Mexico, consiste de 203 especies, incluyendo 42 anuros, 17 caudados,

un cocodrilido, 137 escamosos, y seis tortugas. Catalogamos la distribucion de las especies entre cuatro

regiones fisiograficas aqui reconocidas. El numero total de especies varia de 77 en la Altiplanicie Mexicana,

a 166 en la Sierra Madre Oriental. Las especies individualmente ocupan de una a cuatro regiones (x = 2.1).

Aproximadamente 69% de la herpetofauna de Hidalgo se encuentra en una o dos de las cuatro regiones, lo

Correspondence. ramibautistaa@gmail.com (ARB), uhernndez3@gmail.com (UHS), cruzelizalde@gmail.com (RCE),

christianberriozabal@gmail.com (CBD), izraa.miaral50911@gmail.com (IML), dominic.desantis@gcsu.edu (DLD), jjohnson@utep.edu (JDJ),

eligarcia_18&@hotmail.com (EGP), vmata@utep.edu (VMS), bufodoc@aol.com (LDW)

Amphib. Reptile Conserv. 63 March 2020 | Volume 14 | Number 1 | e224

Herpetofauna of Hidalgo, Mexico

cual es de importancia considerable para su conservacion. El mayor numero de especies que ocupan una sola

region, se encuentra en la Sierra Madre Oriental (25), seguida por las Tierras Costeras del Golfo (15), la Faja

Volcanica Transmexicana (6), y la Altiplanicie Mexicana (2). Una matriz de coeficiente de similitud biogeografica

(CSB), indica que la Sierra Madre Oriental y las Tierras Costeras del Golfo comparten la mayoria de las especies

(72) debido a su proximidad geografica y al numero significativo de especies generalistas presentes en las

Tierras Costeras del Golfo de Mexico, sur de Estados Unidos, Centroameérica, y/o Suramérica. Con respecto a la

superficie, las dos regiones mas grades de Hidalgo, la Sierra Madre Oriental y la Faja Volcanica Transmexicana

reflejan relaciones opuestas sobre la riqueza especifica (166 vs 77 especies, respectivamente) debido a las

caracteristicas ecologicas entre estas. Un dendrograma de similitud basado en el Metodo por Agrupamiento

de Pares no Ponderado con Media Aritmética (MAPMA) revela dos agrupamientos; uno entre la Sierra Madre

Oriental y la Tierras Costeras del Golfo, y el otro entre las dos regiones que comparten la Faja Volcanica

Transmexicana. El primer grupo se debe a que las dos regiones comparten un numero significativo de especies

que ocurren en las tierras costeras del golfo de Norteamérica y Centroameérica, asi como algunas especies que

llegan a Sudamerica. El segundo grupo se debe al contacto de estas dos regiones y sus similitudes ecologicas.

Con respecto a las categorias de distribucion, el mayor numero de especies esta representado por las especies

endemicas a Mexico (104 de 203), seguido por las especies no endémicas (92), endémicas para el estado

(cuatro), y las no nativas (tres). Las principales amenazas ambientales para la herpetofauna de Hidalgo son la

deforestacion, ganaderia, carreteras, contaminacion de fuentes de agua, factores culturales, y enfermedades.

Calculamos el estatus de conservacion de las especies nativas por medio de los sistemas de SEMARNAT (NOM-

059), IUCN, y el EVS, de los cuales el EVS resulto ser el mas util. También utilizamos el metodo de Prioridad

Herpetofaunistica Relativa (PHR) para designar el rango de orden de importancia de las regiones fisiograficas y

determinamos que los valores mas altos pertenecen a la Sierra Madre Oriental. Examinamos las caracteristicas

de las cinco areas protegidas en Hidalgo y determinamos que la mayoria de estas se encuentran en la Faja

Volcanica Transmexicana, que es la segunda region mas importante desde la perspectiva de conservacion.

Tambien determinamos que solamente 78 de las 200 especies nativas registradas en Hidalgo, se encuentran

en estas cinco areas protegidas. Finalmente, establecemos un conjunto de conclusiones y recomendaciones

para la futura proteccion de la herpetofauna de Hidalgo.

Palabras Claves. Anuros, caudados, estatus de conservacion, cocodrilidos, regiones fisiograficas, areas protegidas,

recomendaciones de proteccion, escamosos, tortugas

Citation: Ramirez-Bautista A, Hernandez-Salinas U, Cruz-Elizalde R, Berriozabal-Islas C, Moreno-Lara |, DeSantis DL, Johnson JD, Garcia-Padilla

E, Mata-Silva V, Wilson LD. 2020. The herpetofauna of Hidalgo, Mexico: composition, distribution, and conservation status. Amphibian & Reptile

Conservation 14(1) [General Section]: 63-118 (e224).

[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: 17 March 2019; Accepted: 5 November 2019; Published: 26 March 2020

“One reason we ’ve failed to recognize the damage we’re —_ informacion/hgo/). The 2015 population figure was

doing we’ve assumed it's fine to use our atmosphere as —_ 2,858,359, ranking the state as the 17" most populous

an open sewer.” (http://cuentame.inegi.org.mx/monografias/informacion/

—A] Gore (2017) hgo/). The population density of Hidalgo is 140 people/

km’, ranking 8" in the country (http://cuentame.inegi.

Introduction org.mx/monografias/informacion/hgo/).

The highest elevation in the state of 3,380 m is that of

Hidalgo is an eastern Mexican state located at the | Cerrola Pefiuela, located in the southeastern-most corner of

confluence of four major physiographic regions in the —_ Hidalgo in the municipality of Almoloya (http://cuentame.

country, i.e., the Gulf Coastal Plain, occupying a small _—inegi.org.mx/monografias/informacion/hgo/). Even though

eastern portion of the state; the Sierra Madre Oriental, | much of the state of Hidalgo is mountainous, the heights

extending across the majority of the eastern sector of the — of these mountains are much less imposing than those of

state; the Trans-Mexican Volcanic Belt, traversing the — the neighboring state of Puebla (see Woolrich-Pifia et al.

southern portion of the state; and the Mexican Plateau, 2017). The highest mountain in Puebla, Pico de Orizaba, is

occupying the central and northwestern regions of the 5,747 m in elevation, which is 1.7 times the height of Cerro

state (Fig. 1). Hidalgo 1s bounded to the north by SanLuis _la Pefiuela. Two of the other highest mountains in Mexico

Potosi and Querétaro, to the east by Veracruz and Puebla, _—_ also are partially located in Puebla (Woolrich-Pifia et al.

to the south by Tlaxcala, and to the west by México. 2017). Thus, Hidalgo would be expected to have a smaller

With a surface area of 20,813 km’, Hidalgo is total herpetofauna than Puebla and also support fewer

the 26" largest state in Mexico among the 31 that are | endemic species, both at the national and state levels. An

recognized (http://cuentame.inegi.org.mx/monografias/ examination of these hypotheses is undertaken in this paper.

Amphib. Reptile Conserv. 64 March 2020 | Volume 14 | Number 1 | e224

Ramirez-Bautista et al.

-99.744

-99.19

0510 20 30 40

aa eee Km

Fig. 1. Physiographic regions of Hidalgo, Mexico. Abbreviations: GCL = Gulf Coastal Lowlands; SMO = Sierra Madre Oriental;

MXP = Mexican Plateau; TMV = Trans-Mexican Volcanic Belt.

Materials and Methods

Our Taxonomic Position

We follow the same taxonomic position in this paper

as explained in previous works on other portions of

Mesoamerica (Johnson et al. 2015a,b; Mata-Silva et al.

2015; Teran-Juarez et al. 2016; Woolrich-Pifia et al. 2016,

2017; Nevarez-de los Reyes et al. 2016; Cruz-Saenz

et al. 2016; Gonzalez-Sanchez et al. 2017; Lazcano et

al. 2019). Johnson et al. (2015a) can be consulted for a

detailed statement of this position, with special reference

to the subspecies concept.

System for Determining Distributional Status

The same system developed by Alvarado-Diaz et al.

(2013) for the herpetofauna of Michoacan was employed

here to ascertain the distributional status of members of

the herpetofauna of Hidalgo. Subsequently, 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-Sanchez et al. (2017),

Gonzalez-Sanchez et al. (2017), and Lazcano et al.

(2019) also used this system, which consists of the

following four categories: SE = endemic to Hidalgo; CE

= endemic to Mexico; NE = not endemic to Mexico; NN

= non-native in Mexico.

Amphib. Reptile Conserv.

-98.636

TAsicrrs Madre Oriental =

™ Mexican Plateau

Trans-Mexican Volcanic

Belt R

19.962 20.346

19.579

Systems for Determining Conservation Status

To assess the conservation status of the herpetofauna of

Hidalgo, this survey employed the same systems (_.e.,

SEMARNAT, IUCN, and EVS) as used by 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-

Sanchez et al. (2017), Gonzalez-Sanchez et al. (2017),

and Lazcano et al. (2019). Detailed descriptions of these

three systems appear in the earlier papers of this series

(e.g., Alvarado-Diaz et al. 2013; Johnson et al. 2015a;

Mata-Silva et al. 2015) and do not need to be repeated

here.

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., providing an examination of the composition,

physiographic distribution, and conservation status of

the herpetofauna of a given Mexican state or group of

states. Two years later, the MCS continued with papers

March 2020 | Volume 14 | Number 1 | e224

Herpetofauna of Hidalgo, Mexico

on the herpetofauna of Oaxaca (Mata-Silva et al. 2015)

and Chiapas (Johnson 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 in 2017, and one on Coahuila was

published recently (Lazcano et al. 2019). Thus, this paper

on the herpetofauna of Hidalgo is the eleventh entry in

this series.

Physiography and Climate

Physiographic Regions

The distribution of the herpetofauna of Hidalgo 1s

analyzed using the classification system of physiographic

regions (= physiographic provinces) of INEGI (2000)

and CONABIO (2008). According to these studies, these

consist of four regions, which are briefly described below.

Gulf Coastal Lowlands (GCL). This province belongs

to the Neotropical Region (Morrone 2001) and extends

from the San Fernando River in the state of Tamaulipas

to the Candelaria River located in the Yucatan Peninsula.

This region covers the northern portion of the states of

Quintana Roo, Campeche, Tabasco, and Veracruz, and

small portions of Tamaulipas, eastern San Luis Potosi,

northeastern Hidalgo, northern Puebla, northeastern

Oaxaca, and Campeche. Specifically, in the state of

Hidalgo, this physiographic region is located in the

municipalities of San Felipe Orizatlan and Huejutla

(Sanchez-Rojas and Bravo-Cadena 2017). This region is

located between 25°52’17.02”N, -94°04711.48’°W, and

20°55’36.56’N, -90°18’06.7”W, at elevations spanning

18-1,200 m (Espinosa et al. 2008). Mean annual

precipitation varies between 1,000 and 2,000 mm, while

the average annual temperature for this physiographic

province is 21.2 °C. Dominant vegetation types include

tropical evergreen forest, scrub, subdeciduous forest, and

tropical dry forest (CONABIO 2008).

Sierra Madre Oriental (SMO). This region is located

parallel to the Gulf coastal region of Mexico, which is

connected to the Central Plateau and the Trans-Mexican

Volcanic Belt. The SMO belongs to the Neotropical Realm

and embraces 2.84% of the country (Morrone 2001;

CONABIO 2008). This province is composed mostly of

sedimentary and metamorphic rocks from the Cretaceous

and Jurassic, which makes this province a complex area

from a geological perspective (CONABIO 2008). The

SMO encompasses part of southern Zacatecas, central

and eastern Jalisco, southern Michoacan, Querétaro, and

northeastern Hidalgo (CONABIO 2008). In the state of

Hidalgo, this province extends into the municipalities

Amphib. Reptile Conserv.

of Huehuetla and Tenango de Doria, Calnali, Molango,

Tlanchinol, Lolotla, © Chapulhuacan, Pisaflores,

Tepehuacan, Xochicoatlan, and Eloxochitlan (CONABIO

2008; Ramirez-Bautista et al. 2014; Sanchez-Rojas and

Bravo-Cadena 2017). The northern extent of the SMO lies

at 25°36’23.13”N, -100°17°38.99”"W, and the southern

limit lies at 17°28’45.86”N, -96°04’34.85”W. Elevation

within the SMO in Hidalgo ranges from 100-—3,300 m

(CONABIO 2008; Sanchez-Rojas and Bravo-Cadena

2017). Mean annual precipitation varies considerably,

ranging from 400-800 mm in the montane cloud forests

of the northern region of the municipality of Tlanchinol

(Ramirez-Bautista et al. 2014). The temperature in the

montane environments ranges from 4—28 °C, and from

10-40 °C in the temperate valleys during winter and

summer, respectively. The average annual temperature

is 17.4 °C. On the wet slopes, the dominant vegetation

communities are coniferous forest (28%), oak forest

(26%), and cloud forest (8%); and the vegetation 1s

represented by xerophilous scrub (16%) in the dry region

(CONABIO 2008).

Trans-Mexican Volcanic Belt (TMV). The TMV belongs

to the Neotropical Region (Morrone 2001; CONABIO

2008), and is a volcanic arc located in the central part

of Mexico. The TMV has an east-west orientation,

extending from the state of Veracruz (Gulf of Mexico)

to the state of Nayarit (Pacific Ocean; Ferrusquia-

Villafranca 2007; CONABIO 2008). This belt is formed

by a set of volcanoes of different ages, from Miocene

to Plio-Pleistocene, aligned within 19°31754.81°N,

-98°37°42.45”W and 21°53’40.02”N, -105°36’09.80”"W.

The region occupies 8% of Mexico’s surface area,

ranging in elevation from 1,000—5,700 m. In Hidalgo

the TMV reaches a high point of 2,004 m. Within the

TMV lies the Sierra de Pachuca, which includes the

municipalities of Mineral del Monte, Mineral El Chico,

Huasca de Ocampo, Atotonilco El Grande, and a portion

of Tulancingo (Ramirez-Bautista et al. 2014). The

mean annual precipitation varies from 581—2,236 mm,

and the mean annual temperature is 15.3 °C (Suarez-

Mota et al. 2014). Natural vegetation communities are

represented primarily by coniferous forest (31%) and oak

forest (28%), with the remainder composed of pastures,

subalpine scrub, cloud forest, and farmland. Arid portions

of the region are dominated by xerophilous scrub, while

sub-humid areas contain tropical dry forest.

Mexican Plateau (MXP). This region is within the more

inclusive Nearctic Region (Morrone 2001; CONABIO

2008). This plateau extends through the central zone

of Mexico between 1,700-4,000 m, and it is located

between the Sierra Madre Occidental and Sierra Madre

Oriental. Portions of the MXP fall within the boundaries

of Chihuahua, Coahuila, Durango, Guanajuato,

Hidalgo, Jalisco, Michoacan, Puebla, San Luis Potosi,

Tlaxcala, and Zacatecas. This region is confined within

March 2020 | Volume 14 | Number 1 | e224

Ramirez-Bautista et al.

No. 1. Craugastor decoratus (Taylor 1942). The Adorned

Robber Frog 1s distributed from southern Tamaulipas, eastern

San Luis Potosi, adjacent northern Querétaro, northern

Hidalgo, and adjacent central Veracruz and northern Puebla,

Mexico (Frost 2019). This individual was found at La

Gargantilla, in the municipality of Pisaflores. Wilson et al.

(2013b) calculated its EVS as 15, placing it in the lower portion

of the high vulnerability category. Its conservation status has

been considered as Vulnerable by the IUCN, and as Special

Protection (Pr) by SEMARNAT. Photo by Daniel Lara-Tufiiio.

No. 3. Drvophytes euphorbiaceus (Gunther 1858). The

Southern Highland Treefrog ranges from the “highlands of

southern Mexico (central Veracruz, eastern Hidalgo, Tlaxcala,

and southeastern Puebla to mountains of Oaxaca, including

those south of Oaxaca City)” (Frost 2019). This individual was

encountered at San Mateo, in the municipality of Acaxochitlan.

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 Near Threatened by

the IUCN, but this species is not listed by SEMARNAT. Photo

by Uriel Herndndez-Salinas.

Amphib. Reptile Conserv.

No. 2. Charadrahyla taeniopus (Gunther 1901). The Porthole

Treefrog occurs “on the Atlantic slopes of the Sierra Madre

Oriental from north-eastern Hidalgo southward through

northern Puebla to central Veracruz, Mexico” (Frost 2019).

This individual was located at San Mateo, in the municipality

of Acaxochitlan. Wilson et al. (2013b) assessed its EVS as

13, placing it at the upper limit of the medium vulnerability

category. Its conservation status has been judged as Vulnerable

by the IUCN, and it is placed in the Threatened (A) category by

SEMARNAT. Photo by Uriel Hernandez-Salinas.

No. 4. Dryophytes plicatus (Brocchi 1877). The Ridged

Treefrog is distributed in the Sierra Madre Oriental and the

Cordillera Volcanica along the southern edge of the Mexican

Plateau (Michoacan, Morelos, México, D.F., Tlaxcala, Puebla,

Veracruz, and Hidalgo) [Frost 2019]. This individual was found

at Agua Zarca, in the municipality of Tenango de Doria. Wilson

et al. (2013b) calculated its EVS as 11, placing it in the middle

portion of the medium vulnerability category. Its conservation

status has been considered as Least Concern by the IUCN, and

SEMARNAT lists this treefrog as Threatened (A). Photo by

Uriel Hernandez-Salinas.

March 2020 | Volume 14 | Number 1 | e224

Herpetofauna of Hidalgo, Mexico

24°39’53.31”N, -101°54’04.92”W and 19°31°54.81”N,

-98°37°42.45”W, encompassing a total surface area

of 601,882 m’. The climate is dry, arid, and relatively

cold; the mean annual temperature 1s 18.5 °C, with a

nocturnal mean of -5 °C and a diurnal mean of 25 °C.

Dominant vegetation communities include thorn scrub

and xerophilous scrub. Mean annual precipitation 1s

< 200 mm across all vegetation communities. The

MXP is situated in the central and western portions

of Hidalgo, in the municipalities of Mineral de

la Reforma, Actopan, Ixmiquilpan, Zimapan, and

Huichapan (Ramirez-Bautista et al. 2014).

Climate

Temperature. The monthly minimum, mean, and

maximum temperatures for a single locality for each of

the four recognized physiographic regions in Hidalgo

are shown in Table 1. The elevations for these localities

vary from 420 m at Huehuetla in the GCL to 2,530 m at

Tepeapulco in the TMX.

The mean annual temperature is highest at Huehuetla

(elevation 420 m) in the GCL at 21.2 °C, followed by

Zimapan (elevation 1,763 m) in the MXP at 18.5 °C,

and Tlanchinol (elevation 1,700 m) in the SMO at 17.4

°C, with the lowest mean temperature of 15.3 °C at

Tepeapulco (elevation 2,530 m) in the TMV.

In the four physiographic regions in Hidalgo, the

minimum annual temperatures range from 11.0-16.1

°C lower than the maximum annual temperatures (Table

1). The mean minimum monthly temperatures peak

during May and reach their lowest levels in December or

January. The mean maximum monthly temperatures are

highest in May or July and are lowest in January (Table

1). The mean monthly temperatures are highest in May

and lowest in January (Table 1).

Precipitation. As expected, monthly precipitation is

lowest during the dry season in either December or

February, and highest during the rainy season in either

July or September (Table 2). The data in Table 2 indicate

that 79.2—-94.3% of the annual precipitation occurs during

the rainy season from May to October. Annual rainfall

varies from 537.5 mm on the Mexican Plateau to 1,790.7

mm in the Gulf Coastal Lowlands (Table 2).

Recent Literature on the Hidalgo Herpetofauna

Historically, most information on the herpetofauna of

the state was derived from regional studies or those from

specific localities (see Ramirez-Bautista et al. [2014]

and Lemos-Espinal and Dixon [2016] for a detailed list

of previous studies in the state). In 2010, a checklist

of the state’s herpetofauna (Ramirez-Bautista et al.

2010) included a total of 173 species (54 amphibians

and 119 reptiles), and subsequently Ramirez-Bautista

et al. (2014) published an updated checklist with 183

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Amphib. Reptile Conserv. 68 March 2020 | Volume 14 | Number 1 | e224

Ramirez-Bautista et al.

Table 2. Monthly and annual precipitation data (in mm) for the physiographic regions of Hidalgo, Mexico. Localities for each of

the regions and their elevations are as follows: Gulf Coastal Plain—Huehuetla (420 m); Sierra Madre Oriental—Tlanchinol (1,700

m); Trans Mexican Volcanic Belt—Tepeapulco (2,530 m); Mexican Plateau—Zimapan (1,763 m). Data taken from http://www.

worldclim.org/bioclim (accessed 19 March 2018). The shaded area indicates the months of the rainy season.

Physiographic

Region

Sierra Madre

Oriental

Trans Mexican 103

Volcanic Belt

species (53 amphibians and 130 reptiles), along with

the first comprehensive analysis of the state’s species

richness, diversity, and distribution, and an evaluation

of the conservation status and potential threats to the

persistence of Hidalgo’s herpetofauna. Lemos-Espinal

and Smith (2015) provided an additional checklist of the

herpetofauna of Hidalgo with a total of 175 species, eight

fewer than the number reported by Ramirez-Bautista et

al. (2014). Lemos-Espinal and Dixon (2016) reported the

same figure (175 species). Finally, in 2017 two chapters

on the herpetofauna of the state were published in a book

entitled Biodiversidad del Estado de Hidalgo. The chapter

on amphibians by Goyenechea Mayer-Goyenechea et al.

(2017) listed 53 species, and the chapter on reptiles by

Manriquez-Moran et al. (2017) listed 130 species; both

were the same numbers given by Ramirez-Bautista et al.

(2014). The assessment here provides the most current

species composition for the state following the latest

taxonomic changes.

Gulf Coastal

Lowlands

Composition of the Herpetofauna

Families

The herpetofauna of Hidalgo is represented by 37

families, including nine anuran, three salamander, one

crocodylian, 22 squamate, and two turtle families (Table

3). This total figure is 62.7% of the 59 herpetofaunal

families recorded from Mexico (Wilson et al. 2013a,b).

No caecilians are registered in the state. Of the amphibian

families, 86.4% of the species are placed in the families

Bufonidae, Eleutherodactylidae, Hylidae, Ranidae, and

Plethodontidae (Tables 4—5). Among the reptile families,

75.7% of the species are allocated to the families

z me [oe [oe [

Anguidae, Dactyloidae, Phrynosomatidae, Colubridae,

Dipsadidae, Natricidae, and Viperidae (Tables 4—5).

Genera

Ninety-six herpetofaunal genera are represented in

Hidalgo, including 20 anuran, seven salamander, one

crocodylian, 65 squamate, and three turtle genera

(Table 3). These 96 taxa comprise 45.7% of the 210

genera recorded from Mexico (Wilson et al. 2013a,b).

Among the amphibians, the most species-rich genera

are Craugastor (five species), Eleutherodactylus (five),

Lithobates (five), and Chiropterotriton (nine). Among

the reptile genera, the most speciose are Sce/oporus (13),

Thamnophis (nine), Crotalus (nine), and Norops (five).

Species

The herpetofauna of Hidalgo is composed of 203 species,

including 42 anurans, 17 salamanders, one crocodylian,

137 squamates, and six turtles (Table 3). The current

numbers of native species in these five groups in Mexico

are, respectively, 248, 151, three, 865, and 51 (J. Johnson,

unpub.). The 203 species in Hidalgo comprise 15.4% of

the 1,318 species in the entire Mexican herpetofauna (J.

Johnson, unpub., 26 June 2019).

The one state sharing a common border with Hidalgo

examined thus far in the Mexican Conservation Series is

Puebla (Woolrich-Pifia et al. 2017), the herpetofauna of

which consists of 267 species or 1.3 times the richness of

Hidalgo (203). This comparative figure somewhat resembles

the relative areas of the two states. The surface area of

Puebla is 34,306 km? (Woolrich-Pifia et al. 2017) and that of

Hidalgo, as noted above, is 20,813 km?; thus, Puebla is 1.6

Table 3. Composition of the native and non-native herpetofauna of Hidalgo, Mexico.

Order Families

Anura 9

Caudata 3

Subtotal 12

Crocodylia 1

Squamata 22

Testudines 2

Subtotal 25

Total 37

Amphib. Reptile Conserv.

Genera Species

20 42

7 17

2a, 59

1 1

65 137

3 6

69 144

96 203

March 2020 | Volume 14 | Number 1 | e224

Herpetofauna of Hidalgo, Mexico

No. 5. Rheohyla miotympanum (Cope, 1863). The Small-eared

Treefrog occupies the “highlands of Nuevo Leon and Coahuila

(Sierra Madre Oriental) to Guanajuato (Sierra Santa Rosa),

Hidalgo, and Oaxaca, adjacent Veracruz, and central Chiapas

in eastern and central Mexico” (Frost 2019). This individual

was located at San Mateo, in the municipality of Acaxochitlan.

Wilson et al. (2013b) determined its EVS as 9, placing it at the

upper limit of the low vulnerability category. Its conservation

status has been considered as Near Threatend by the IUCN,

but this species is not listed by SEMARNAT. Photo by Uriel

Herndndez-Salinas.

No. 7. Lithobates johni (Blair, 1965). John’s Frog is distributed

from “southeastern San Luis Potosi, eastern Hidalgo, and

northern Puebla, Mexico” (Frost 2019). This individual was

found at Rio Blanco, in the municipality of Huehuetla. Wilson

et al. (2013b) calculated its EVS as 14, placing it at the lower

limit of the high vulnerability category. Its conservation status

has been considered as Endangered by the IUCN, and it is

placed in the Endangered (P) category by SEMARNAT. Photo

by Christian Berriozabal-Islas.

Amphib. Reptile Conserv.

No. 6. Sarcohyla robertsorum (Taylor 1940). Roberts’ Treefrog

occupies “the Sierra Madre Oriental in eastern Mexico (Puebla

and Hidalgo)” (Frost 2019). This individual was encountered in

Zoquizoquiapan, in the municipality of Metztitlan. Wilson et al.

(2013b) assessed its EVS as 13, placing it at the upper limit of

the medium vulnerability category. Its conservation status has

been considered as Endangered by the IUCN, and it is placed in

the Threatened (A) category by SEMARNAT. Photo by Raquel

Herndndez-A ustria.

No. 8. Lithobates montezumae (Baird 1854). The Montezuma

Leopard Frog ranges from San Luis Potosi, Querétaro, Jalisco,

and eastern Durango south to the southeastern edge of the

Mexican Plateau in Tlaxcala, Puebla, Hidalgo, Ciudad de

México, and Veracruz, Mexico (Frost 2019). This individual was

discovered at E] Huemac, in the municipality of Tezontepec de

Aldama. Wilson et al. (2013b) estimated 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

the IUCN, and it is allocated to the Special Protection (Pr)

category by SEMARNAT. Photo by Christian Berriozabal-

Islas.

March 2020 | Volume 14 | Number 1 | e224

Ramirez-Bautista et al.

Table 4. Distribution of the amphibians, crocodylians, squamates, and turtles of Hidalgo, Mexico, by physiographic region.

Abbreviations: SMO = Sierra Madre Oriental; MXP = Mexican Plateau; TMV = Trans-Mexican Volcanic Belt; and GCL = Gulf

Coastal Lowlands. See text for descriptions of these regions. * = species endemic to Mexico; ** = species endemic to Hidalgo; and

*** = non-native species.

eo or ee

| Anura(42speciesy |

| Bufonidae(6species) | CT

Sarcohyla bistincta*

Sarcohyla charadricola*

Sarcohyla robertsorum*

i |

Scinax staufferi

Smilisca baudinii

Tlalocohyla picta

Trachycephalus vermiculatus

Leptodactylidae (2 species)

Eleutherodactylus longipes*

Amphib. Reptile Conserv. 71 March 2020 | Volume 14 | Number 1 | e224

Herpetofauna of Hidalgo, Mexico

Table 4 (continued). Distribution of the amphibians, crocodylians, squamates, and turtles of Hidalgo, Mexico, by physiographic

region. Abbreviations: SMO = Sierra Madre Oriental; MXP = Mexican Plateau; TMV = Trans-Mexican Volcanic Belt; and GCL =

Gulf Coastal Lowlands. See text for descriptions of these regions. * = species endemic to Mexico; ** = species endemic to Hidalgo;

and *** = non-native species.

po SMO MXP TMV | Gct |

| Ambystomatidae (I species) |

| Ambystomavelasci* | 8

| Plethodontidae (15 species) |

| Aquiloeurycea cephalica* | tT

| Bolitoglossa platydactyla* |

| Chiropterotriton arboreus* | HT

| Chiropterotriton chico** LT

| Chiropterotriton chiropterus* |

| Chiropterotriton chondrostega* | tT

| Chiropterotriton dimidiatus** | #8

| Chiropterotriton terrestris** | dT

|Isthmurabellii* 8

|Isthmuragigantea* |

| Pseudoeuryceaaltamontana*® | TE

| Pseudoeurycealeprosa* | 8

| Salamandridae(I species) |

| Notophthalmus meridionalis_— | dT

| Crocodylia(I species) |

| Crocodylidae(I species) |

| Crocodylusmoreletii_ |

| Squamata (136 species) |

| Anguidae(Sspecies) |

| Abroniataeniata* TT 8

| Barisiaimbricata* CT 8

| Gerrhonotusinfernalis |

| Gerrhonotusliocephalus |

| Gerrhonotus ophiurus* |

| Corytophanidae(3 species) |

| Basiliscusvittatus TC

Corytophanes hernandezii SSS EE

| Laemanctus serratus LT

| Dactyloidae(Sspecies) |

| Noropslaeviventris |

| Noropslemurinus TT

| Noropsnaufragus* |

| Noropspetersii CE UT

| Noropssericeus CT

| Dibamidae(I species) |

| Anelytropsispapillosus* |

| Eublepharidae(1 species) |

| Coleonyxelegans |

| Gekkonidae(I species) |

| Hemidactylus frenatus*** |

| Iguanidae(I species) |

| Clenosauraacanthura LT

| Phrynosomatidae(14 species) |

" +

| Sceloporus bicanthalis* |

| Sceloporuscyanogenys, |

| Sceloporus grammicus dL 8

HTH

EGG

Amphib. Reptile Conserv. 72 March 2020 | Volume 14 | Number 1 | e224

Ramirez-Bautista et al.

Table 4 (continued). Distribution of the amphibians, crocodylians, squamates, and turtles of Hidalgo, Mexico, by physiographic

region. Abbreviations: SMO = Sierra Madre Oriental; MXP = Mexican Plateau; TMV = Trans-Mexican Volcanic Belt; and GCL =

Gulf Coastal Lowlands. See text for descriptions of these regions. * = species endemic to Mexico; ** = species endemic to Hidalgo;

and *** = non-native species.

Xantusiidae (4 species)

Lepidophyma occulor*

Lepidophyma sylvaticum*

Lepidophyma flavimaculatum

Lampropeltis ruthveni*

Leptophis diplotropis*

Leptophis mexicanus

Aspidoscelis gularis

Masticophis flagellum

Conopsis lineata*

Ficimia hardyi*

Xenosauridae (3 species)

Masticophis mentovarius

Amphib. Reptile Conserv.

;

March 2020 | Volume 14 | Number 1 | e224

Herpetofauna of Hidalgo, Mexico

Table 4 (continued). Distribution of the amphibians, crocodylians, squamates, and turtles of Hidalgo, Mexico, by physiographic

region. Abbreviations: SMO = Sierra Madre Oriental; MXP = Mexican Plateau; TMV = Trans-Mexican Volcanic Belt; and GCL =

Gulf Coastal Lowlands. See text for descriptions of these regions. * = species endemic to Mexico; ** = species endemic to Hidalgo;

and *** = non-native species.

Number of

regions occupied

po SMO MXP TMV | Gct |

| Salvadorabairdi* CT

| Salvadora grahamiae_ | UT 8

| Senticolistriaspis LT 8

| Spilotespullatus TT

| Tantillabocourti* CT

| Tantillarubra

| Trimorphodontau* LT

| Dipsadidae(27 species) |

| Adelphicos quadrivirgatum | tT

| Amastridium sapperi_ Lt

| Chersodromus rubriventris* |

| Coniophanesfissidens LT

| Coniophanesimperialis |

| Coniophanespiceivittis |

| Diadophis punctatus | 8

| Geophis latifrontalis* |

| Geophislorancai CL

| Geophis mutitorques* | 8

| Geophis semidoliaus* | 8

| Geophis turbidus*

| Hypsiglenajani CL

| Hypsighenatanzeri* LT

| Imantodescenchoa | UT

| Imantodes gemmistratus |

| Leptodeiramaculata* | UT

| Leptodeira septentrionalis | UT

| Niniadiademata |

| Pliocercuselapoides | UT

| Rhadinaeadecorata |

| Rhadinaeagaigeae* |

| Rhadinaeahesperia® |

| Rhadinaea marcellae* |

| Rhadinaea quinguelineata* |

| Sibonnebulams

| Tropidodipsas sartorii LT

| Elapidae(2species) |

| Micrurusdiastema TT

| Micrurustener CT 8

| Leptotyphlopidae(3 species) |

| Epictiawynni®

|Renadulcis

| Renamyopica* CT

| Natricidae(13 species) |

| Nerodiarhombifer LT

| Storeriadekayi TT

| Storeriahidalgoensis* |

| Storeria storerioides* |

| Thamnophis cyrtopsis dL 8

| Thamnophiseques | U8

| Thamnophis marcianus | HT 8

| Thamnophis melanogaster* | HT

| Thamnophis proximus Lt =

| Thamnophis pulchrilatus* | Tt

| Thamnophis scalaris* | UT

Taxa Physiographic regions of Hidalgo

+)4+]}+]4+]+]4+]+

NP Re l[RIW]wlwoldy

+/+]+]/+]+/+]+]+]4+]+ +]+]+ |+

ALA

+ 2

+]+]4+]4

WIN /|BRlelTelTele Jp tb

+]+]+

NPWPRWPOTNT Rel ely

Amphib. Reptile Conserv. 74 March 2020 | Volume 14 | Number 1 | e224

Ramirez-Bautista et al.

Table 4 (continued). Distribution of the amphibians, crocodylians, squamates, and turtles of Hidalgo, Mexico, by physiographic

region. Abbreviations: SMO = Sierra Madre Oriental; MXP = Mexican Plateau; TMV = Trans-Mexican Volcanic Belt; and GCL =

Gulf Coastal Lowlands. See text for descriptions of these regions. * = species endemic to Mexico; ** = species endemic to Hidalgo;

and *** = non-native species.

i 2 as SES

al:

phlopidae (1 species)

Indotyphlops braminus*** a

Viperidae (13 species)

times the size of Hidalgo. Therefore, the state area/species

richness ratio for Hidalgo is 102.5 compared to 128.5 for

Puebla.

Patterns of Physiographic Distribution

Here, four physiographic regions in Hidalgo are

recognized (Fig. 1), and the occurrence of the members

of the herpetofauna among these four regions are

documented in Table 4 and summarized in Table 5.

The total numbers of species in each of these four

regions vary from a low of 77 in the Mexican Plateau

(MXP) to a high of 166 in the Sierra Madre Oriental

(SMO). The intermediate figures are 85 for the Trans-

Mexican Volcanic Belt and 95 for the Gulf Coastal

Lowlands. Interestingly, the number of species recorded

from the Sierra Madre Oriental is about 1.7 to 2.2

times those in the three other regions in the state. The

herpetofauna of the SMO comprises 81.8% of that of the

entire state (203 species).

As expected, the largest proportions of the species

by broader herpetofaunal groups are found in the SMO

(Table 5), including 36 of 42 anurans (85.7%), 12 of 17

Amphib. Reptile Conserv.

Number of

regions occupied

———]

salamanders (70.6%), 114 of 137 squamates (83.2%),

and four of six turtles (66.7%).

As noted above, the numbers of species in the other

three regions are approximately half of that found in

the SMO (Table 5). Of these three regions, the largest

number of species (95) is found in the Gulf Coastal

Lowlands, including 22 of 42 anurans (52.4%), two

of 17 salamanders (11.8%), one of one crocodylian

(100%), 67 of 137 squamates (48.9%), and three of six

turtles (50.0%). The next largest number of species in

these three regions (85) is found in the Trans-Mexican

Volcanic Belt, including 20 of 42 anurans (47.6%), 14 of

17 salamanders (82.4%), 49 of 137 squamates (35.8%),

and two of six turtles (33.3%). Finally, the smallest

number of species (77) is registered on the Mexican

Plateau, including 18 of 42 anurans (42.9%), seven of 17

salamanders (41.2%), 51 of 137 squamates (37.2%), and

one of six turtles (16.7%).

The members of the Hidalgo herpetofauna inhabit

from one to four of the four physiographic regions, as

follows: one (48; 23.6%); two (93; 46.0%); three (59;

29.2%); and four (three; 1.5%). The average regional

March 2020 | Volume 14 | Number 1 | e224

Herpetofauna of Hidalgo, Mexico

ae ON LORRI RES

No. 9. Ambystoma velasci (Dugés 1888). The Plateau Tiger

Salamander ranges from northwestern Chihuahua south along

the eastern slope of the Sierra Madre Occidental and southern

Nuevo Leon to Hidalgo in the Sierra Madre Oriental, west to

Zacatecas, and south into the Transverse Volcanic range of

central Mexico (Frost 2019). This individual was located in

Parque Nacional El Chico, in the municipality of Mineral del

Chico. Wilson et al. (2013b) determined its EVS as 10, placing

it at the lower limit of the medium vulnerability category. Its

conservation status has been considered as Least Concern by

the IUCN, and it has been placed in the Special Protection (Pr)

category by SEMARNAT. Photo by Christian Berriozabal-

Islas.

No. 11. Bolitoglossa platydactyla (Gray 1831). The Broad-

footed Salamander occurs from “southern Tamaulipas and

eastern San Luis Potosi south through Hidalgo to southern

Veracruz, Puebla, Oaxaca, and extreme northeastern Chiapas,

Mexico” (Frost 2019). This individual was found at Cececamel

in the municipality of San Felipe Orizatlan. Wilson et al. (2013b)

ascertained its EVS as 15, placing it in the lower portion of the

high vulnerability category. Its conservation status had been

judged as Near Threatened by the IUCN, and SEMARNAT

has placed it in the Special Protection (Pr) category. Photo by

Cristian Raul Olvera-Olvera.

Amphib. Reptile Conserv.

No. 10. Aquiloeurycea cephalica (Cope 1865). The Chunky

False Brook Salamander is distributed in “the Transverse

Volcanic Range in Ciudad de México and states of Veracruz,

Hidalgo, México, Puebla, and Morelos” (Frost 2019). This

individual was found in the municipality of Tlanchinol. Wilson

et al. (2013b) assessed its EVS as 14, placing it at the lower

limit of the high vulnerability category. Its conservation status

has been considered as Near Threatened by the IUCN, and as

occupying the Threatened (A) category by SEMARNAT. Photo

by Uriel Hernandez-Salinas.

~ .

~ N.

4 :

s te

Se a eg a nee

No. 12. [sthmura belli (Gray 1850). Bell’s Salamander is found

in “southern Tamaulipas, Tlaxcala, Hidalgo and the Sierra

Madre del Sur of Guerrero, Mexico, and west and north to

southern Nayarit and southern Zacatecas” (Frost 2019). This

individual was encountered at Las Coas, in the municipality

of Tlahuiltepa. 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

Vulnerable by the IUCN, and as Threated (A) by SEMARNAT.

Photo by Christian Berriozabal-Islas.

March 2020 | Volume 14 | Number 1 | e224

Ramirez-Bautista et al.

Table 5. Summary of distribution occurrence of herpetofaunal families in Hidalgo, Mexico, by physiographic province. See Table

4 for explanation of abbreviations.

species

SMO

| Bufonidag

eee eer, eel

| Eleutherodactylidae TS

N}yn

| | [nena |

[Total 8

| Crispi Hi

[STS aN (00 9) [a | a ee

PT ES Se ee Ee ee Ee Ee Ss

| Corytophanidae BZ 8

LDaciviendas 2 | SS eS | ee

|Wibarbidact I eee eee ee

|Eublepharidae

Ce | ae ay [a eed ed (a ee

DEES (A) (QS eg (Eee

Semmeidag?) 1 | ee ea ae ee

I I

ape PR ye | ni Go] nf

|Subtotal

|SumTotal 0G | 8S HT

]

occupancy is 2.1, which means that, on average, each Charadrahyla taeniopus*

individual species occupies only about half of the Scaphiopus couchii

physiographic regions found in the state. Isthmura gigantea*

A sizable proportion of the herpetofauna is distributed Norops laeviventris

in one or two regions (141, or 69.5% of the total). This Coleonyx elegans

proportion if very close to that seen for Puebla (68.5%; Sceloporus cyanogenys

Woolrich-Pifia et al. 2017); in the case of Puebla, Plestiodon tetragrammus

however, there are six regions instead of the four found Xenosaurus mendozai*

in Hidalgo. Xenosaurus newmanorum*

The number of species found in a single region range Xenosaurus tzacualtipantecus*

from three (in the MXP) to 25 (in the SMO). The 25 Masticophis flagellum

single-region species in the SMO are: Pituophis catenifer

Tantilla rubra

Amphib. Reptile Conserv. 77 March 2020 | Volume 14 | Number 1 | e224

Herpetofauna of Hidalgo, Mexico

Table 6. Pair-wise comparison matrix of Coefficient of Biogeographic Resemblance (CBR) data of herpetofaunal relationships for

the four physiographic regions in Hidalgo, 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 Fig. 6 for the UPGMA dendrogram produced from the

CBR data.

Sierra Madre Nietitan Plateau Trans-Mexican Gulf Coastal

Oriental Volcanic Belt Lowlands

Sierra Madre

Oriental

Mexican

Plateau

Trans-Mexican

Volcanic Belt

Gulf Coastal

Lowlands

Geophis latifrontalis*

Geophis lorancai*

Geophis turbidus*

Hypsiglena tanzeri*

Rhadinaea decorata

Rhadinaea marcellae*

Micrurus diastema

Storeria dekayi

Storeria hidalgoensis*

Metlapilcoatlus nummifer*

Ophryacus smaragdinus*

Terrapene mexicana*

Fourteen of the 25 SMO single-region species (56.0%)

are Mexican endemics (indicated by asterisks); the

remainder are non-endemic species.

The 15 single-region species in the GCL are:

Craugastor berkenbuschii*

Scinax staufferi

Bolitoglossa platydactyla*

Crocodylus moreletii

Basiliscus vittatus

Hemidactylus frenatus***

Ctenosaura acanthura

Coluber constrictor

Leptophis diplotropis*

Coniophanes imperialis

Coniophanes piceivittis

Epictia wynni*

Scaphiodontophis annulatus

Trachemys venusta

Kinosternon scorpioides

Four of the 15 GCL single-region species are Mexican

endemics (single asterisk), one is a non-native (triple

asterisk), and the remainder are non-endemic species.

The six single-region species in the TMV are:

Amphib. Reptile Conserv.

Chiropterotriton chico**

Chiropterotriton magnipes*

Pseudoeurycea altamontana*

Sceloporus bicanthalis*

Rhadinaea gaigeae*

Rhadinaea hesperia*

Five of the six TMV single-region species are Mexican

endemics and one is a state endemic.

The two single-region species in the MXP are:

Lampropeltis annulata

Rena dulcis

Both of these species are non-endemics.

In summary, of the 48 single-region species in

Hidalgo, 23 are Mexican endemics, one is a State

endemic, 23 are non-endemics, and one is a non-native.

Of the four physiographic regions, the SMO 1s of the

greatest conservation importance given that it houses

the greatest overall number of species (166), the greatest

number of single-region species (25), and the largest

number of country endemics (14).

A Coefficient of Biogeographic Resemblance (CBR)

matrix was created for studying the herpetofaunal

similarity relationships among the four physiographic

regions in Hidalgo (Table 6) and those data were used

to construct a UPGMA dendrogram. The SMO contains

the greatest species richness (166 species) and the MXP

the least (77 species). The mean species richness value

for all four areas is 105.5. The number of shared species

between each of the regional pairs ranges from a high

of 72 between SMO and GCL to a low of 13 between

TMV and GCL. The mean value of shared species

among all four regions is 47.8. The lowest number

of shared species between the TMV and the GCL (13

Species) was expected because these two regions are

situated on opposite ends of Hidalgo, are not connected

geographically (being completely separated by the

March 2020 | Volume 14 | Number 1 | e224

Ramirez-Bautista et al.

= a

No. 13. /sthmura gigantea (Taylor 1939). The Giant False

Brook Salamander ranges “in the La Joya-Jalapa region of

Veracruz and into northeastern Hidalgo, Mexico” (Frost

2019). This individual was encountered at Chilijapa, in the

municipality of Tepehuacan de Guerrero. Wilson et al. (2013b)

determined its EVS as 16, placing it in the middle portion of

the high vulnerability category. Its conservation status has

been considered as Critically Endangered by the IUCN, but

this species is not listed by SEMARNAT. Photo by Christian

Berriozabal-Islas.

No. 15. Gerrhonotus ophiurus Cope 1867. This alligator lizard

occurs in the Mexican states of Hidalgo, Veraruz, San Luis

Potosi, Querétaro, Michoacan, and Puebla (Ramirez-Bautista

et al. 2014). This individual was encountered at El Demafii,

in the municipality of Tlahuiltepa. Wilson et al. (2013a)

assessed 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 the IUCN, but this species is

not listed by SEMARNAT. Photo by Christian Berriozabal-

Islas.

Amphib. Reptile Conserv.

No. 14. Barisiaimbricata(Wiegmann, 1828). The Transvolcanic

Alligator Lizard ranges in the Trans-Mexican Volcanic Belt

and the Sierra Madre Oriental, in the states of México, Ciudad

de México, Querétaro, Hidalgo, Jalisco, Puebla, Oaxaca,

Michoacan, Morelos, and Tlaxcala. This individual was found

at Puentecillas in the municipality of Singuilucan. Wilson et al.

(2013b) calculated its EVS as 15, placing it in the lower portion

of the high vulnerability category. Its conservation status has

been assessed as Least Concern by the IUCN, and it has been

placed in the Special Protection (Pr) category by SEMARNAT.

Photo by Cristian Raul Olvera-Olvera.

he ?

No. 16. Norops naufragus (Campbell, Hillis, and Lamar 1989).

The Hidalgo Anole is found only in the states of Hidalgo and

Puebla in Mexico (Ramirez-Bautista et al. 2014). This individual

was found at Cuatatlan, in the muncipality of Tlanchinol. Wilson

et al. (2013a) ascertained its EVS as 13, placing it at the upper

limit of the medium vulnerability category. Its conservation

status has been considered as Vulnerable by the IUCN, and it is

placed in the Special Protection (Pr) category by SEMARNAT.

Photo by Christian Berriozabal-Islas.

March 2020 | Volume 14 | Number 1 | e224

Herpetofauna of Hidalgo, Mexico

SMO and the MXP), and are environmentally different

on an elevational scale. The GCL, with an elevational

range from near sea level to 1,200 m, contains tropical

evergreen forest and subhumid formations of scrublands

to tropical dry forests. On the other hand, the TMV

with a limited geographic area within Hidalgo contains

humid, semihumid, and subhumid vegetation in montane

environments at elevations from 1,000 m in large sloping

river valleys to 3,400 m on volcanic peaks. The SMO

and the GCL share the most species (72), which also was

not unexpected because they are directly adjacent to each

other in Hidalgo, and the tropical lowland environments

of the GCL ascend into the mountainous habitats of the

SMO. The pairwise comparisons of regions aligned in

order from highest to lowest species richness (underlined

values) and their corresponding numbers of shared

species (in parentheses) are:

SMO 166: GCL (72), MXP (67), TMV (66)

GCL 95: SMO (72), MXP (16), TMV (13)

TMV 85: SMO (66), MXP (53), GLC (13)

MXP 77: SMO (67), TMV (53), GLC (16)

In general, the pattern indicates how species richness

values within each of the four biogeographic regions

of Hidalgo equate to numbers of shared species among

the other three regions. There is a higher correlation

of species richness values to number of shared species

between regions that are in contact with each other, but

also observed correlations between regions that share

similar ecological parameters. Interestingly, the two

regions that share the most species (72) are a highland

region (SMO) and a lowland region (GCL), which is

probably due to the GCL containing many generalist

species that can endure both montane and non-montane

environments in low to moderate elevations. The fact that

the GCL shares few species with the MXP and the TMV

gives credibility to the premise that regions separated by

ecological barriers will share fewer species than they will

with regions in direct contact.

The following data show ranges and mean numbers of

shared species (bold in parentheses) for each of the four

regions that are arranged according to increasing species

richness (underlined values) in each region:

Sierra Madre Oriental — SMO (166): 66—72 (68.3)

Gulf Coastal Lowlands — GCL (95): 13-72 (33.6)

Trans-Mexican Volcanic Belt — TMV (85): 13-66 (44.0)

Mexican Plateau — MXP (77): 16-67 (45.3)

The mean numbers of shared species compared to the

species richness values in all four regions indicate that

higher species richness in pairwise comparisons does

not translate into higher reciprocal numbers when all

regional pairs are totaled. The most apparently extreme

example of this is the comparison between the SMO

Amphib. Reptile Conserv.

and the GCL—which are 1“ and 2" in species richness,

but 1° and last (4) in mean numbers of shared species,

respectively. The SMO also has higher mean numbers

of shared species with TMV (44.0) and MXP (45.3),

but if GCL (2"4 in species richness, last in mean number

of shared species) 1s removed from the tabulation, the

three montane regions have even higher mean numbers

of shared species. Specifically, the average number of

shared species between the SMO, the TMV, and the MXP

combined is 62.0 (calculated from Table 6).

Regarding area, the two largest geographic regions,

the SMO and the MXP, reflect opposite relationships

in species richness (166 vs. 77 species, respectively).

The SMO contains more tropical, subtropical humid,

and semihumid vegetation formations compared to the

mostly subhumid environments in the MXP, in addition

to being in direct contact with the second most species-

rich region, the GCL, which shares the highest number

of species in Hidalgo with the SMO. The GCL, the

second most speciose region and third smallest region in

the state, contains 10 more species than does the TMV,

the smallest area by far that also contains less humid

and semi-humid environments than does GCL. Also

note that Hidalgo is a relatively small state in area (5"

smallest of the 31 in Mexico), which undoubtedly affects

species richness. As an example, the adjacent state of

Puebla, which is slightly larger and contains two more

physiographic regions than does Hidalgo, contains 267

species of amphibians and reptiles (Woolrich-Pifia et al.

2017).

Based on the data in Table 6, a UPGMA dendrogram

(Fig. 6) was created to depict the herpetofaunal similarity

resemblance patterns in a hierarchical fashion among the

four physiographic regions of Hidalgo (see map, Fig. 1).

The dendrogram is composed of two distinct clusters;

one comprising two montane regions (MXP and TMV)

at the 0.65 level and the other containing one montane

region (SMO) and the lowland region (GCL) at the 0.55

level. The two clusters connect together at the 0.39 level.

Regions within both clusters are adjacent to each other

and depict patterns of ecological similarity; and in the case

of the SMO and the GCL, they share generalist species

that primarily occur on the Gulf-facing side that ascends

from the lowlands (the GCL) into the higher elevations

in the SMO. Fifty-three of the 203 herpetofaunal species

(26.2%) presently known from Hidalgo are shared only

between the SMO and the GLC (Table 4), and many of

them are wide-ranging species along the Gulf versant of

Mexico, some of which also enter the USA, and/or Central

America and South America (Wilson and Johnson 2010).

Those 53 species also represent 73.6% of the 72 species

shared among the SMO, the GCL, and other regions in

Hidalgo. We also predict that other species now restricted

to either the SMO or especially the GCL eventually will

be discovered in both regions. In our opinion, the shared

generalist species within the SMO and the GCL are the

exclusive reason why the SMO clusters with the GCL

instead of with the two other montane regions (MXP and

TMV).

March 2020 | Volume 14 | Number 1 | e224

Ramirez-Bautista et al.

A GV ie

No. 17. Phrynosoma_ orbiculare (Linnaeus 1758). The

Mountain Horned Lizard is known from the states of

Chihuahua, Aguascalientes, Hidalgo, Querétaro, San Luis

Potosi, Michoacan, Ciudad de México, Estado de México,

Jalisco, Morelos, Tlaxcala, and Guanajuato (Ramirez-Bautista

et al. 2014). This individual was located in Parque Nacional

El Chico, in the municipality of Mineral del Chico. Wilson et

al. (2013a) determined 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 the IUCN,

and as Threatened (A) by SEMARNAT. Photo by Christian

Berriozabal-Islas.

No. 19. Sceloporus minor Cope 1885. The Minor Scaly Lizard

ranges into the states of Nuevo Leén, Zacatecas, San Luis

Potosi, Tamaulipas, Querétaro, and Guanajuato (Ramirez-

Bautista et al. 2014). This individual was located at La Mesa,

in the Municipality of Zacualtipan. Wilson et al. (2013a)

calculated its EVS as 14, placing it at the lower limit of the

high vulnerability category. Its conservation status has been

considered as Least Concern by the IUCN, but this species is

not listed by SEMARNAT. Photo by Aaron Garcia-Rosales.

Amphib. Reptile Conserv.

No. 18. Sceloporus bicanthalis Smith 1937. The Transvolcanic

Bunchgrass Lizard is distributed in the states of Hidalgo,

México, Oaxaca, Puebla, and Veracruz (Ramirez-Bautista et al.

2014). This individual was found in the municipality of Mineral

El Chico. 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

the IUCN, but this species is not listed by SEMARNAT. Photo

by Uriel Herndndez-Salinas.

No. 20. Lepidophyma occulor Smith 1942. The Jalpan Tropical

Night Lizard has a restricted distribution in adjacent areas of

Querétaro, San Luis Potosi, and Hidalgo (Ramirez-Bautista

et al. 2014). This individual came from Puerto Oscuro, in the

municipality of Pisaflores. Wilson et al. (2013a) determined its

EVS as 14, placing it at the lower limit of the high vulnerability

category. Its conservation status has been considered as Least

Concern by the IUCN, and it is placed in the Special Protection

(Pr) category by SEMARNAT. Photo by Daniel Lara-Tufifio.

March 2020 | Volume 14 | Number 1 | e224

Herpetofauna of Hidalgo, Mexico

Distribution Status Categorizations

The assessment of the distribution status of the members

of the Hidalgo herpetofauna here uses the system

developed by Alvarado-Diaz et al. (2013) and employed

in all the other entries in the Mexican Conservation

Series (see above). The categories in the system are

The numbers of species in each of the four categories,

in decreasing order of size, are: country endemics, 104

(51.2%); non-endemics, 92 (45.3%); state endemics,

four (2.0%); and non-natives, three (1.5%). As with

the states of Michoacan (Alvarado-Diaz et al. 2013),

Nayarit (Woolrich-Pifia et al. 2016), Jalisco (Cruz-Saenz

et al. 2017), and Puebla (Woolrich-Pifia et al. 2017), the

non-endemic, country endemic, state endemic, and non-

native, and data are presented in Table 7 and summarized

in Table 8.

greatest number of herpetofaunal species in Hidalgo

lies within the country endemic category. The largest

number falls within the non-endemic category in the

Table 7. Distributional and conservation status measures for members of the herpetofauna of Hidalgo, Mexico. Distributional

Status: SE = endemic to Hidalgo; 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 categorizations: 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. See text for

explanations of the EVS, IUCN, and SEMARNAT rating systems.

Environmental

Vulnerability

Category (Score)

IUCN

categorization

SEMARNAT

status

Distributional

status

Amphib. Reptile Conserv. 82 March 2020 | Volume 14 | Number 1 | e224

Ramirez-Bautista et al.

Table 7 (continued). Distributional and conservation status measures for members of the herpetofauna of Hidalgo, Mexico.

Distributional Status: SE = endemic to Hidalgo; 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 categorizations: 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. See text

for explanations of the EVS, IUCN, and SEMARNAT rating systems.

Distributional | =2¥ironmental IUCN SEMARNAT

Vulnerability a fae

Status Category (Score) categorization Status

Lithobates montezumae*

Species

Amphib. Reptile Conserv. 83 March 2020 | Volume 14 | Number 1 | e224

Herpetofauna of Hidalgo, Mexico

Table 7 (continued). Distributional and conservation status measures for members of the herpetofauna of Hidalgo, Mexico.

Distributional Status: SE = endemic to Hidalgo; 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 categorizations: 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. See text

for explanations of the EVS, IUCN, and SEMARNAT rating systems.

Distributional | =2¥ironmental IUCN SEMARNAT

Vulnerability sled.

status Category (Score) categorization Status

Species

Amphib. Reptile Conserv. 84 March 2020 | Volume 14 | Number 1 | e224

Ramirez-Bautista et al.

Table 7 (continued). Distributional and conservation status measures for members of the herpetofauna of Hidalgo, Mexico.

Distributional Status: SE = endemic to Hidalgo; 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 categorizations: 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. See text

for explanations of the EVS, IUCN, and SEMARNAT rating systems.

Distributional | =2¥ironmental IUCN SEMARNAT

Vulnerability ie

status Category (Score) categorization status

Species

Amphib. Reptile Conserv. 85 March 2020 | Volume 14 | Number 1 | e224

Herpetofauna of Hidalgo, Mexico

Table 7 (continued). Distributional and conservation status measures for members of the herpetofauna of Hidalgo, Mexico.

Distributional Status: SE = endemic to Hidalgo; 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 categorizations: 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. See text

for explanations of the EVS, IUCN, and SEMARNAT rating systems.

Environmental

Vulnerability

Category (Score)

[Rhadinaeageigeae® +i ce [a [pp | ___ns__

[Rhadinaea quinguelieaa® | ce | __s)_ [pp |___mr_

IUCN SEMARNAT

categorization status

Distributional

Species statue

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Ramirez-Bautista et al.

Table 7 (continued). Distributional and conservation status measures for members of the herpetofauna of Hidalgo, Mexico.

Distributional Status: SE = endemic to Hidalgo; 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 categorizations: 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. See text

for explanations of the EVS, IUCN, and SEMARNAT rating systems.

Distributional | Environmental IUCN SEMARNAT

Species Vulnerability die A

status categorization status

Category (Score)

Ophryacus smaragdinus*

Terrapene mexicana*

Trachemys venusta

other states surveyed thus far: Oaxaca (Mata-Silva et — Principal Environmental Threats

al. 2015); Tamaulipas (Teran-Juarez et al. 2016); Nuevo

Leon (Nevarez-de los Reyes et al. 2016); and Chiapas __In this section we highlight the problems that we view as

(Johnson et al. 2015a). most significantly affecting the sustainability of Hidalgo’s

In the ten previous individual-state entries inthe MCS, — herpetofauna populations. The major threats include the

the numbers of state endemics varied considerably, from —_ increasing and unregulated clearing of forests for farming

one in Nayarit and Nuevo Leon (Woolrich-Pifia et al. and livestock, construction of roads, the constant and

2016; Nevarez-de los Reyes et al. 2016) toa maximum __ increasing pollution of water bodies, emerging diseases,

of 93 in Oaxaca (Mata-Silva et al. 2015). The number —_and strongly ingrained cultural factors (Ramirez-Bautista

of state endemics in Hidalgo is near the lower end ~ el al. 2014; Cruz-Elizalde et al. 2017).

of that range at four, and all of them are plethodontid

salamanders in the genus Chiropterotriton: C. chico, C. Deforestation

dimidiatus, C. mosaueri, and C. terrestris (Table 7).

As noted in the introduction, we hypothesized that the = The state of Hidalgo encompasses 903,502.5 ha used for

number of endemic species should be greater for the state — livestock and agricultural activities (INEGI 2011). The

of Puebla than for Hidalgo. Woolrich-Pifia et al. (2017) — area utilized for these activities, however, continues to

reported the number of country endemics for Puebla as —_ increase, consequently eliminating ~1,200-—5,102 ha of

162 (60.7% of state total). As noted above, this figure natural vegetation cover per year (SEMARNAT 2012).

for Hidalgo is 104, which is 51.2% of the state total. The | Hidalgo ranks from fifth to seventh among the 31 Mexican

number of state endemics, however, is the same for these __ states in terms of deforestation rates (SEMARNAT 2012).

two states, at four (Woolrich-Pifia et al. 2017), which | Unfortunately, the loss of natural habitats affects both

supports our hypothesis since Puebla has the greater biological communities and human development in the

total number of endemic species (166) than does Hidalgo —_ region, since deforestation accelerates the loss of soils,

(108). increases water runoff, and accelerates the evaporation

Three non-native species occur in Hidalgo: _ rates of water bodies that serve many local communities.

Lithobates catesbeianus, Hemidactylus frenatus, and At the local level, increasing deforestation is driven

Indotyphlops braminus. Two of these three (H. frenatus primarily by agriculture. After farmers clear an area

and J. braminus) are the most widespread of the non- (often on pronounced slopes), they typically only use

native species thus far recorded in the 11 entries in the — this land for one or two years. Once the terrain loses

Mexican Conservation Series (Woolrich-Pifia et al. most of its top soil layers to erosion (Fig. 1), the site is

2017), having been recorded, as of this paper, in 10 and abandoned in favor of clearing of a new area of native

11 states, respectively. vegetation. For instance, in the SMO large trees are

cut down and the lower vegetation is eliminated with

Amphib. Reptile Conserv. 87 March 2020 | Volume 14 | Number 1 | e224

Herpetofauna of Hidalgo, Mexico

oat

No. 21. Lepidophyma sylvaticum Taylor 1939. The Madrean

Tropical Night Lizard is distributed in the Mexican states of

Puebla, Hidalgo, Nuevo Leon, Querétaro, San Luis Potosi,

Tamaulipas, and Veracruz (Ramirez-Bautista et al. 2014).

This individual was located at La Cueva, in the municipality

of Pisaflores. 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 the IUCN, and it is placed in the Special Protection

(Pr) category by SEMARNAT. Photo by Daniel Lara-Tufifio.

> OR -_

No. 23. Xenosaurus newmanorum Taylor 1949. Newman’s

Knob-scaled Lizard ranges from southeastern San Luis Potosi

and extreme northern Hidalgo (Ramirez-Bautista et al. 2014).

This individual was found at La Ameca, in the municipality of

Pisaflores. Wilson et al. (2013a) assessed its EVS as 15, placing

it in the lower portion of the high vulnerability category. Its

conservation status has been considered as Endangered by the

IUCN, and is allocated to the Special Protection (Pr) category

by SEMARNAT. Photo by Christian Berriozabal-Islas.

Amphib. Reptile Conserv.

No. 22. Xenosaurus mendozai Nieto Montes de Oca, Garcia

Vazquez, Zufiga-Vega, and Schmidt-Ballardo 2013. This

individual was found at El Pinalito, in the municipality of

Jacala de Ledezma. Wilson et al. (2013a) calculated its EVS

as 15, placing it in the lower portion of the high vulnerabilty

category. Its conservation status has not been determined by the

IUCN, and this species is not listed by SEMARNAT. Photo by

Christian Berriozabal-Islas.

No. 24. Xenosaurus tzacualtipantecus Woolrich-Pifia and

Smith 2012. The Zacualtipan Knob-scaled Lizard is limited to

the Sierra Madre Oriental in the states of Hidalgo and Veracruz

(Ramirez-Bautista et al. 2014). This individual came from

La Mojonera, in the municipality of Zacualtipan. Wilson et

al. (2013a) calculated its EVS as 17, placing it in the middle

portion of the high vulnerability category. Its conservation

status has been considered as Near Threatened by the IUCN,

but this species is not listed by SEMARNAT. Photo by Lia

Victoria Berriozabal-Varela.

March 2020 | Volume 14 | Number 1 | e224

Ramirez-Bautista et al.

Fig. 2. Gulf Coastal Lowlands. Riparian vegetation in the

vicinity of Achiquihurxtla in the municipality of Atlapexco.

Photo by Cristian Raul Olvera-Olvera.

Fig. 4. Mexican Plateau. Vegetation in the vicinity of Santa

Monica in the municipality of Metztitlan. Photo by Cristian

Raul Olvera-Olvera.

controlled fires (Fig. 2). The removal of the arboreal

layer affects a diversity of species that are dependent

on specific microclimatic parameters. Some examples

of herpetofaunal taxa involved are salamanders of the

genera Aquiloeurycea, Chiropterotriton, and Isthmura,

anurans of the genera Craugastor, Eleutherodactylus,

Charadrahyla, and Plectrohyla, lizards of the genera

Abronia, Norops, | Corytophanes, | Laemanctus,

Lepidophyma, and Xenosaurus, and snakes of the genera

Boa, Spilotes, Metlapilcoatlus, Bothrops, and Ophryacus

(Cruz-Elizalde et al. 2017).

Livestock

Similar to agricultural deforestation, livestock ranching

also involves vegetation removal for short-term

exploitation. Livestock activities are associated with

the destruction of thousands of ha of pristine forest. The

soils in these pastures are prone to erosion and can only

support one or two years of cattle grazing. Ranchers are

then forced to look for new sites to clear at the expense

of the natural ecosystems (Ramirez-Bautista et al. 2014).

As a testimony to this crisis, in regions such as SMO

and GCL, pasturelands have increased dramatically.

Originally, these areas were covered with cloud forests

and tropical forests (Fig. 3). The semiarid region in the

state is not exempt from deforestation either, and goats

are the main concern. Goat herders take their animals

Amphib. Reptile Conserv.

Fig. 3. Sierra Madre Oriental. Panoramic view in the vicinity

of Diego Mateo inside the Parque Nacional El Chico in the

municipality of Mineral del Chico. Photo by Cristian Raul

Olvera-Olvera.

ir,

Fig. 5. Trans-Mexican Volcanic Belt. Sierra de Pachuca.

Xerophilic scrub in Cerro de San Cristobal in the municipality

of Pachuca. Photo by Paola Lazcano-Judrez.

to feed in areas covered with shrubs, destroying the

slow-growth plants such as cacti and agaves, and in turn

leading to erosion of the fragile soil (Ramirez-Bautista et

al. 2014; Magno-Benitez et al. 2016).

Roads

Road infrastructure 1s important for the economic and

social growth in the state. However, as is becoming more

evident, this development brings adverse consequences

to biodiversity (Puc Sanchez et al. 2013). Specifically,

roads act as physical barriers for many amphibian

and reptile species and reduce connectivity between

populations. Vehicle-induced mortality or “roadkill”

is one of the most visible effects of roads, as many

herpetofauna cross busy roadways due to migration or

March 2020 | Volume 14 | Number 1 | e224

Herpetofauna of Hidalgo, Mexico

Fig. 6. A UPGMA generated dendrogram illustrating the

similarity relationships of species richness among. the

herpetofauna in the four physiographic regions of Hidalgo

(based on the data in Table 6). Similarity values were calculated

values using Duellman’s (1990) Coefficient of Biogeographic

Resemblance (CBR).

dispersal, or use paved roads for basking (Figs. 4-5).

Pollution of water bodies

Continuous human population growth in Hidalgo

(SEMARNAT 2012) and the lack of urban development

plans have exacerbated the improper disposal of waste

products and, consequently, have affected water sources

such as rivers in the MXP, SMO, and GCL. Additionally,

many fields used to produce vegetables in the western

region of the state have been irrigated with sewage water,

which, unfortunately, contaminates the soils and local

water sources. The sewage water that ends up in rivers

has also modified the water properties significantly,

causing many frog and turtle populations to disappear

from those sites (Ramirez-Bautista et al. 2014; Magno-

Benitez et al. 2016).

Myths and other cultural factors

Two cultural aspects that contribute to the detriment of

Hidalgo’s herpetofauna are the lack of understanding

regarding the important roles of amphibians and reptiles

in ecosystems, and harmful misconceptions that often

lead to direct persecution (Cruz-Elizalde et al. 2017).

For instance, many people in Hidalgo believe that some

species of salamanders (genera Aquiloeurycea and

Chiropterotriton) and lizards (genera Abroniaand Barisia)

are venomous, while all snakes are indiscriminately

regarded as dangerous, and, therefore, killed on sight.

Additionally, many people believe that the salamanders

Aquiloeurycea_ cephalica, Bolitoglossa_platydactyla,

Chiropterotriton arboreus, C. chondrostega, and the

snake Pituophis deppei somehow impregnate women;

therefore, encounters with these creatures frequently

Amphib. Reptile Conserv.

Fig. 7. Forest fires. A forest fire for land use conversion in the

vicinity of El Naranjal, in the municipality of Pisaflores. Photo

by Christian Berriozabal-Islas.

Fig. 8. Deforestation for ranching purposes. Cattle pasture

in the municipality of Tepehuacan de Guerrero. Photo by

Christian Berriozabal-Islas.

end up with them being killed (Ramirez-Bautista et al.

2014; Cruz-Elizalde et al. 2017). Consumption of most

herpetofaunal members has not been well-documented

in the state. In some rural communities, however, the

salamander Ambystoma velasci is known to be part of

the diet (Fig. 6) and rattlesnakes are used as part of the

folk medicine by some inhabitants (Cruz-Elizalde et al.

201):

Diseases

Globally, many amphibian populations are disappearing

due to chytridiomycosis, caused by the fungus

Batrachochytrium dendrobatidis (Bd, Skerrat et al. 2007).

Unfortunately, this disease was reported recently in

Hidalgo in the anurans Craugastor rhodopis, Lithobates

berlandieri, L. johni, and Rheohyla miotympanum

(Hernandez-Austria 2017). Two factors have been

identified as the main drivers of the successful spread of

this infection in Hidalgo: the exotic American Bullfrog

(L. catesbeianus) and global climate change (Kriger et al.

2006). Monitoring of this infection is necessary in order

to assess its impact on the diverse native frog populations

(Hernandez-Austria 2017).

Under these circumstances, government authorities

March 2020 | Volume 14 | Number 1 | e224

Ramirez-Bautista et al.

Fig. 9. Deforestation for agricultural purposes. Change in land

use for agricultural purposes in the vicinity of San Cristobal in the

municipality of Metztitlan. Photo by Cristian Raul Olvera-Olvera.

by locals for maintenance in captivity as a pet in El Borbollon in

the municipality of Huehuetla. Photo by Christian Berriozabal-

Islas.

at all levels and conservation groups must invest more

effort in the protection of these species and the habitats

where they are found. These efforts are particularly

critical in regions that harbor species and habitats that are

already vulnerable to anthropogenic stressors. Another

critical step is that the respective authorities need to

invest more resources in the continuous education of the

general public on the importance of the herpetofauna of

the state. Otherwise, adequate protection of these species

will always remain an elusive goal.

Conservation Status

The conservation status of the members of the

herpetofauna of Hidalgo is assessed here using the

same three systems of conservation assessment as in the

previous entries in the Mexican Conservation Series (see

above). These systems are those of SEMARNAT (2010),

the IUCN Red List (http://tucnredlist.org), and the EVS

(Wilson et al. 2013a,b), and these three systems have

been updated as necessary.

The SEMARNAT System

The SEMARNAT system is a means of conservation

Amphib. Reptile Conserv.

Fig. 10. Invasive species. Lithobates catesbeianus in the

vicinity of El Naranjal in the municipality of San Felipe

Orizatlan. Photo by Cristian Raul Olvera-Olvera.

~ & :

Fig. 12. Urbanization. Urban growth in the vicinity of Molango

de Escamilla in the municipality of the same name. Photo by

Cristian Raul Olvera-Olvera.

status assessment developed and implemented by the

Secretaria del Medio Ambiente y Recursos Naturales of

the federal government of Mexico (SEMARNAT 2010).

The ratings are available for some of the herpetofaunal

species inhabiting Hidalgo as shown in Table 7 and

summarized in Table 9. Three categories of assessment

exist in the SEMARNAT system: Endangered (P),

Threatened (A), and Under Special Protection (Pr);

and the species remaining unassessed in this system are

assigned a “No Status” (NS) category.

The data in Table 9 indicate that of the 200 native

species in Hidalgo, only 93 (45.8%) have been evaluated

using this system. This leaves 107 (52.7%) without a

conservation assessment based on SEMARNAT.

If one can assume that SEMARNAT personnel have

placed a greater emphasis on species endemic to Mexico

or some portion thereof (1.e., a single state), then that

consideration should be evident from a comparison of

the assignments to both distributional categories and to

SEMARNAT categories. In order to determine whether

this sort of bias is evident, such comparisons are presented

in Table 10. These data demonstrate that the majority of

the non-endemic species (61 of 92; 66.3%) currently

remain unevaluated in the SEMARNAT system. The

comparable figures are 44 of the 104 (42.3%) country

March 2020 | Volume 14 | Number 1 | e224

Herpetofauna of Hidalgo, Mexico

‘ th “¢ >

a , ren »

No. 25. Conopsis lineata (Kennicott 1859). The Lined

Tolucan Ground Snake occurs in the central Mexican states of

Guanajuato, Guerrero, Jalisco, Estado de México, Michoacan,

Morelos, Oaxaca, Puebla, Queretaro, San Luis Potosi, Tlaxcala,

Veracruz, and Ciudad de México (Ramirez-Bautista et al. 2014).

This individual was found at Puentecillas, in the municipality of

Singuilucan. Wilson (2013a) determined its EVS as 13, placing

it at the upper limit of the medium vulnerability category. Its

conservation status is assessed as Least Concern by the IUCN,

but this species is not listed by SEMARNAT. Photo by Cristian

Raul Olvera-Olvera.

yea mdse

No. 27. Pituophis deppei (Duméril 1853). The Mexican Bull

Snake occurs in the states of Aguascalientes, Chihuahua,

Coahuila, Durango, Guanajuato, Hidalgo, Jalisco, México,

Michoacan, Nuevo Leon, Oaxaca, Puebla, San Luis Potosi,

Querétaro, Tlaxcala, Veracruz, Zacatecas, and Ciudad de

México (Ramirez-Bautista et al. 2014). This individual was

encountered in the municipality of Mineral El Chico. Wilson

et al. (2013a) calculated its EVS as 14, placing it at the lower

limit of the high vulnerability category. Its conservation status

has been considered as Least Concern by the IUCN, and it is

placed in the Threatened (A) category by SEMARNAT. Photo

by Uriel Hernandez-Salinas.

Amphib. Reptile Conserv.

2 pte < s

M ager t am Ne; vY

No. 26. Lampropeltis annulata. Kennicott 1861. The Mexican

Milksnake is distributed in Nuevo Leon, Querétaro, and

Tamaulipas, and perhaps Coahuila, eastern San Luis Potosi, and

Hidalgo (Ruane et al. 2014). This individual was encountered

at Venados, in the municipality of Metztitlan. 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 the IUCN, and this species is not

listed by SEMARNAT. Photo by Cristian Raul Olvera-Olvera.

Pg Mie kB eT Be Fe OE onal

No. 28. Chersodromus rubriventris (Taylor 1949). The Redbelly

Earth Runner “is found in the Sierra Madre Oriental in the

States of San Luis Potosi, Querétaro and Hidalgo” (Canseco-

Marquez et al. 2018: 159). This individual was photographed

at Chilijapa, in the municipality of Tepehuacan de Guerrero.

Wilson et al. (2013a) calculated its EVS as 14, placing it at the

lower limit of the high vulnerability category. Its conservation

status has been considered as Endangered by the IUCN, and it is

placed in the Special Protection (Pr) category by SEMARNAT.

Photo by Christian Berriozabal-Islas.

March 2020 | Volume 14 | Number 1 | e224

Ramirez-Bautista et al.

Table 8. Summary of the distributional status of herpetofaunal families in Hidalgo, Mexico.

Family a eaeeie os

Bufonidae

Craugastoridae

Eleutherodactylidae

Hylidae

Leptodactylidae

Microhylidae

Ranidae

Rhinophrynidae

Scaphiopodidae

Subtotal

Ambystomatidae

Plethodontidae

Salamandridae

Subtotal

Total

Crocodylidae

Subtotal

Anguidae

Corytophanidae

Dactyloidae

Dibamidae

Eublepharidae

Gekkonidae

Iguanidae

Phrynosomatidae

Scincidae

Sphenomorphidae

Teiidae

Xantusiidae

Xenosauridae

Subtotal

Boidae

Colubridae

Dipsadidae

Elapidae

Leptotyphlopidae

Natricidae

Sibynophiidae

Typhlopidae

Viperidae

Subtotal

Emydidae

Kinosternidae

Subtotal

Total

Sum Total

a aN

oe)

—

ioe) ~

ios)

oS)

co)

[SSeS

a ee

—

[ee

a a)

—_

endemics and two of the four state endemics (50.0%).

Although these figures do not indicate a clear bias in favor

of the Mexican endemic species, they do demonstrate

that the SEMARNAT system will not be of much use

in assessing the conservation status of the Mexican

herpetofauna, and specifically the Hidalgo herpetofauna,

Amphib. Reptile Conserv.

Distributional status

Non-endemic Country State Endemic

(NE) Endemic (CE) (SE)

4 2

Non-native

(NN)

until all species are included.

The IUCN System

The IUCN system of conservation assessment is

intended to be applicable to all organisms, although most

of its evaluations are applied to vertebrate animals and

March 2020 | Volume 14 | Number 1 | e224

Herpetofauna of Hidalgo, Mexico

Table 9. SEMARNAT categorizations for herpetofaunal species in Hidalgo, Mexico, arranged by family. Non-native species are

excluded.

Special No status

Endangered (P) Threatened (A) protection (Pr) (NS)

Hylidae

Leptodactylidae

Microhylidae

Ranidae

Rhinophrynidae

Scaphiopodidae

Subtotal

| Plethodontidae |S eB 8

GS Lan | © 5 on [27 | TT, | OOS S| <1)

Crocodylidae

Subtotal

Anguidae

Corytophanidae

Dactyloidae

Dibamidae

Eublepharidae

Iguanidae

| Sphenomorphidae |

foe

|Subtotal CE a a

|Boidaes Te ee

}Colubridane | 8

|Dipsadidae CT a

a a a ee ee eee eee ee

| Leptotyphlopidae ee

|Natricidae TB

|Sibynophiidae | ee

|Viperidae OT 3 a

Subtotal ee Sa RN PER Eas

[Emydidae fe

|Kinostenidae | Ae

|Subtotal CE

|Total CT ee TT

| Sum Total 200 tor

flowering plants. For example, of 67,222 animal species __ reptiles of 10,711; thus, 58.6% of the world’s recognized

assessed, 46,092 are vertebrates (68.6%); and of 24,230 _ reptile species have been assessed by the IUCN; while

plant species evaluated, 22,566 (93.1%) are flowering — the figure for amphibians is 84.4% of 7,832 species

plants IUCN Red List version 2017-3: Tables 3a,b in (Amphibian Species of the World, http://research.amnh.

that list). The vertebrate animal assessments include — org/vz/herpetology/amphibia/; accessed 17 April 2018).

6,609 for amphibians and 6,278 for reptiles IUCN Red — Thus, a significantly greater proportion of amphibian

List version 2017-3: Table 3a). The Reptile Database — species have been assessed than reptile species. For the

website (http://www.reptile-database.org/; accessed 17 global herpetofauna, 12,887 (69.5%) of 18,543 total

April 2018) provides a February 2018 total count for species have been assessed.

—"

Amphib. Reptile Conserv. 94 March 2020 | Volume 14 | Number 1 | e224

Ramirez-Bautista et al.

Table 10. Comparison of SEMARNAT and distributional categorizations.

SSS SEMARNAT category

Threatened)

State-endemic (SE)

Table 11. [UCN Red List categorizations for herpetofaunal families in Hidalgo, Mexico. Non-native species are excluded. The shaded

columns to the left are the “threatened” categories, and those to the right are the categories which indicate that available conservation status

data are too limited to allow the species to be placed in any other IUCN category, or the species has not been evaluated.

Number IUCN Red List categorization

Family of Critically Near Least

species Endangered Endangered | Vulnerable Threatened | Concern

Country-endemic (CE)

| Plethodontidac__|__d

| Total |S

Subtotal

—

ies)

Kinosternidae 4

142

200

Category Total 20

—

35 129 36

Amphib. Reptile Conserv. 95 March 2020 | Volume 14 | Number 1 | e224

Mexico

Herpetofauna of Hidalgo

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March 2020 | Volume 14 | Number 1 | e224

Ramirez-Bautista et al.

No. 29. Jmantodes cenchoa (Linnaeus 1758). The Bluntheaded

Tree Snake is broadly distributed “from Tamaulipas and

Oaxaca, Mexico, southward through much of Central America

to Ecuador, on the Pacific versant, and to Paraguay on the

cisandean side of South America” (Lemos-Espinal and Dixon

2013: 191). This individual was found at Cececamel in the

municipality of San Felipe Orizatlan. Wilson et al. (2013a)

assessed its EVS as 6, placing it in the middle of the low

vulnerability category. Its conservation status has not been

evaluated at the IUCN, but its SEMARNAT status is judged as

Special Protection (Pr). Photo by Cristian Rail Olvera-Olvera.

No. 31. Metlapilcoatlus nummifer (Ruppell 1845). The Mexican

Jumping Viper is found “from San Luis Potosi southward

through Hidalgo and west-central Veracruz to northern and

southeastern Oaxaca (Lemos-Espinal and Dixon 2013: 246).

This individual was found in El Pinalito, in the municipality

of Jacala de Ledezma. Wilson et al. (2013a) estimated its EVS

as 13, placing it at the upper limit of the medium vulnerability

category. Its conservation status is indicated as Least Concern

by the IUCN and as Threatened (A) by SEMARNAT. Photo by

Christian Berriozabal-Islas.

Amphib. Reptile Conserv.

No. 30. Micrurus diastema (Duméril, Bibron, and Duméril

1854). The Variable Coral Snake is found “on the Atlantic

versant from northern Veracruz and northern Oaxaca,

Mexico, to northwestern Honduras” (McCranie 2011: 457).

This individual was photographed at Laguna de Atezca, in

the municipality of Molango de Escamilla. Wilson (2013a)

calculated its EVS as 8, placing it in the upper portion of the

low vulnerability category. Its conservation status has been

assessed as Least Concern by the the IUCN, and this elapid is

listed as a species of Special Protection (Pr) by SEMARNAT.

Photo by Christian Berriozabal-Islas.

No. 32. Boiirons asper (Gas 1884), The Terciopelo

is a wide-ranging pit viper occurring “from southwestern

Tamaulipas, Mexico, to coastal Venezuela on the Atlantic

versant, and from Costa Rica to southern Ecuador on the

Pacific versant, with a disjunct population occurring in

southern Chiapas, Mexico, and adjacent Guatemala” (Lemos-

Espinal and Dixon 2013: 247). This individual was found in

La Esperanza II, in the municipality of Huehuetla. Wilson et

al. (2013a) determined its EVS as 12, placing it in the upper

portion of the medium vulnerability category. Its conservation

status has not been determined by the IUCN, and this species

is not listed by SEMARNAT. Photo by Christian Berriozabal-

Islas.

March 2020 | Volume 14 | Number 1 | e224

Herpetofauna of Hidalgo, Mexico

In previous entries in the Mexican Conservation

Series (e.g., Woolrich-Pifia et al. 2017), the IUCN system

of conservation evaluation has been criticized for several

reasons. Nonetheless, the IUCN system is sufficiently

broadly applied that its comparison here to the other

systems is instructive. Thus, the IUCN categorizations

for the members of the Hidalgo herpetofauna are shown

in Table 7 and summarized in Table 11.

Of 200 native members of the herpetofauna, 164

(82.0%) have been assessed by the IUCN system. This

percentage is similar to that found by Woolrich-Pifia

et al. (2017) for the herpetofauna of the adjacent state

of Puebla (79.5%). Of these 164 species, 35 have been

placed in one of the three IUCN “threat categories:” seven

as CR, 14.as EN, and 14 as VU (Table 11). The seven CR

species are Bromeliohyla dendroscarta, Chiropterotriton

arboreus, C. chiropterus, C. magnipes, C. mosaueri, C.

terrestris, and Isthmura gigantea. Five of these species

are country endemics and two are state endemics;

One is an anuran and six are salamanders. The 14 EN

species are Sarcohyla arborescandens, S. charadricola,

S. robertsorum, Lithobates johni, Chiropterotriton

chondrostega, C. dimidiatus, C. multidentatus,

Pseudoeurycea altamontana, Notophthalmus

meridionalis, Xenosaurus newmanorum, Ficimia hardyi,

Chersodromus rubriventris, Rhadinaea marcellae, and

Thamnophis_ melanogaster, and include 12 country

endemics, one state endemic and one non-endemic. Four

of these species are anurans, five are salamanders, one

is a lizard, and four are snakes. The 14 VU species are

Craugastor decoratus, C. rhodopis, Eleutherodactylus

longipes, E. verrucipes, Charadrahyla taeniopus, Isthmura

bellii, Pseudoeurycea leprosa, Abronia taeniata, Norops

naufragus, Sceloporus megalepidurus, Lepidophyma

gaigeae, Storeria hidalgoensis, Thamnophis scaliger, and

Trachemys venusta; and include 13 country endemics and

one non-endemic. Five of these species are anurans, two

are salamanders, four are lizards, two are snakes, and one is

a turtle. In total, of the 35 species in the IUCN “threatened

categories,” 30 are endemic to Mexico or to Hidalgo

(85.7%); 10 species are anurans, 13 are salamanders, five

are lizards, six are snakes, and one 1s a turtle.

Of the 129 species placed in the IUCN “lower risk

categories” (NT and LC), only seven (5.4%) are allocated

to the NT category; the remaining 122 are placed in the

LC category. The seven NT species are Craugastor

berkenbuschii, Dryophytes euphorbiaceus, Rheohyla

miotympanum, Aquiloeurycea cephalica, Bolitoglossa

platydactyla, Lampropeltis ruthveni, and Kinosternon

herrerai. All seven of these species are country endemics;

three are anurans, two are salamanders, one is a snake,

and one is a turtle.

The 122 LC species comprise 61.0% of the 200 native

species in Hidalgo. Whether such a high proportion

of these species are actually of “Least Concern” 1s

questionable; and these allocations are examined in

detail below.

Amphib. Reptile Conserv.

Thirty-six of the members of the native Hidalgo

herpetofauna have not been placed in either the

“threatened categories” or the “lower risk categories,”

including four allocated to the DD category and 32 to

the NE categories. Inasmuch as these 36 species make up

18.0% of the native herpetofauna, they also are examined

in greater detail in the following section.

The EVS System

The EVS (Environmental Vulnerability Score) system

was developed originally for use in evaluating the

conservation status of the Honduran herpetofauna, but

has since been deployed in the assessment of other

components of the Mexican and Central American

herpetofaunas (Wilson et al. 2010, 2013a,b; and all

entries in the Mexican Conservation Series [see above]).

In the present study, the EVS values for the 200 native

Species are given in Table 7 and summarized in Table 12.

The EVS values range from 3 to 19, which is one

less than the entire theoretical range of 3—20. The most

frequent values (applied to 10 or more species) are 6

(16 species), 8 (14), 9 (14), 10 (17), 11 (16), 12 (17), 13

(26), 14 (18), 15 (17), and 16 (11). These ten values are

applied to 166 of the 200 native species (83.0%). The

lowest score of 3 was calculated for three anuran species

(Rhinella horribilis, Smilisca baudinii, and Scaphiopus

couchii) and the highest score of 19 was calculated for

two turtles (7errapene mexicana and Trachemys venusta).

As in prior MCS studies, the EVS are organized

here into three categories of low, medium, and high

vulnerability. As such, the species counts increase from

low vulnerability (66) to medium vulnerability (76), and

then decrease in the high vulnerability (58) category.

Generally, this pattern is typical of state herpetofaunas

that contain more non-endemic species than country and

state endemics, as was found in Chiapas (Johnson et al.

2015), Tamaulipas (Teran-Juarez et al. 2016), Nuevo

Leon (Nevarez-de los Reyes et al. 2016), and Coahuila

(Lazcano et al. 2019).

When the IUCN categories for the Hidalgo

herpetofauna are compared with those from the EVS

system (Table 13), 35 of the 58 high vulnerability

species (60.3%) are allocated to one of the three IUCN

“threat categories.” This relatively high proportion is due

primarily to the number of amphibians evaluated by the

IUCN as CR, EN, or VU; 23 of 59 amphibian species

(39.0%) are anurans (10 species) or salamanders (13),

compared to 12 of 144 reptiles (8.3%). No squamates,

turtles, or crocodylians are assessed as CR, only five

squamates are assessed as EN, and six squamates and

one turtle are assessed as VU. At the other extreme, the

66 low vulnerability species (by EVS) comprise 54.1%

of the 122 LC species (by IUCN). As demonstrated in

previous MCS entries, the results from the IUCN and

EVS systems do not complement one another very well.

As reported in previous MCS studies, the main

March 2020 | Volume 14 | Number 1 | e224

Ramirez-Bautista et al.

Table 13. Comparison of Environmental Vulnerability Scores (EVS) and IUCN categorizations for members of the herpetofauna of

Hidalgo, Mexico. Non-native species are excluded. Shaded area at the top (EVS scores from 3 to 9) encompasses low vulnerability

category scores, and the shaded area at the bottom (EVS scores from 14 to 19) indicates high vulnerability category scores.

IUCN category

Lo rr Se ee eee ee ee ee ee

| Total | 7 ta aT 8200

reason for the poor correspondence between the

IUCN and EVS systems is that a large proportion

(158 of 200, 79.0%) of the species are assigned to the

NE, DD, and LC categories. Interestingly, the four

DD species are all country endemic snakes (Geophis

latifrontalis, Hypsiglena tanzeri, Rhadinaea gaigeae,

and R. quinquelineata). Three of these four species are

categorized as high vulnerability species, and the fourth

(R. gaigeae) has an EVS of 12 putting it in the medium

vulnerability category (Table 14). As a result, we believe

the conservation needs of these four species are ill-served

by leaving them in the DD category of IUCN. Thus, we

think that the two species with an EVS of 15 (Hypsiglena

tanzeri and Rhadinaea quinquelineata) would be more

appropriately placed in the EN category, the one with an

EVS of 14 (Geophis latifrontalis) in the VU category,

and the one with an EVS of 12 (Rhadinaea gaigeae) in

the NT category.

Thirty-two of the 200 native species (16.0%) have

not been evaluated by the IUCN (Table 15). These 32

species comprise an interesting amalgam of country/

state endemics and non-endemic species. Of the 32

species, 11 are species endemic to Hidalgo (one species)

or to Mexico (11 species). One of the criticisms levelled

against the IUCN system of conservation evaluation is

that it is too slow to keep up with taxonomic innovation

(Johnson et al. 2015). Of the 11 endemic species listed

in Table 15, eight have been described, resurrected

from synonymy, or elevated from subspecies to species

level in the present decade (1.e., Chiropterotriton chico,

Holcosus amphigrammus, Xenosaurus mendozai, X.

tzacualtipantecus, Lampropeltis polyzona, Geophis

lorancai, G. turbidus, Epictia wynni, and Ophryacus

smaragdinus). Of the 20 non-endemic species not yet

assessed by the IUCN, all range into the United States,

Central America or both. Clearly, a more rapidly-

applied system of conservation assessment 1s needed,

especially given the rate at which anthropogenic habitat

modification and destruction occur. As with the DD

categorized species, we believe that the EVS provides a

means for allocating the NE species to IUCN categories.

Thus, we suggest that species with an EVS of 17 or 18

should be placed in the CR category (Chiropterotriton

chico, Xenosaurus tzacualtipantecus, and Crotalus

Table 14. Components of the Environmental Vulnerability Scores (EVS) for members of the herpetofauna of Hidalgo, Mexico, that

are allocated to the IUCN Data Deficient category. * = country endemic; ** = state endemic.

Species Geographic

Distribution

Amphib. Reptile Conserv.

Environmental Vulnerability Score (EVS)

Reproductive

Mode/Degree of

Persecution

Ecological

Distribution

March 2020 | Volume 14 | Number 1 | e224

Herpetofauna of

Hidalgo, Mexico

Table 15. Components of the Environmental Vulnerability Scores (EVS) for members of the herpetofauna of Hidalgo, Mexico,

currently classified as Not Evaluated (NE) by the IUCN. Non-native taxa are excluded. * = country endemic; ** = state endemic.

Species

Geographic

Distribution

Norops sericeus

Ctenosaura acanthura

Sceloporus cyanogenys*

Xenosaurus mendozai*

Xenosaurus tzacualtipantecus*

Boa imperator

Ficimia olivacea*

Holcosus amphigrammus*

Lampropeltis annulata

totonacus), those with an EVS of 15 or 16 in the EN

category (Xenosaurus mendozai and Geophis turbidus),

and those with an EVS of 13 or 14 in the VU category

(Sceloporus cyanogenys, Geophis lorancai, Epictia

wynni, and Ophryacus smaragdinus). The three species

with an EVS of 12 perhaps should be allocated to the NT

category (Ctenosaura acanthura, Lampropeltis annulata,

and Bothrops asper). The remaining species with EVS of

3 to 11 probably should be placed in the LC category.

Previous studies in the MCS series have demonstrated

that the largest proportions of the herpetofaunal species

found in any of the regions examined were allocated to

the LC category by the IUCN. Such is also the case in

this study of the Hidalgo herpetofauna. As noted above,

122 of the 200 native species (61.0%) are in this category

(Table 16), 52 (42.6%) of which are country endemics.

We believe it is unlikely that such a large proportion of

these 122 species are really of “Least Concern.” Based

on the same reasoning employed with the DD and NE

species above, our opinion is that the species with an EVS

Lampropeltis polyzona*

Drymobius margaritiferus

Amphib. Reptile Conserv.

100

Environmental Vulnerability Score (EVS)

Reproductive

Mode/Degree of

Persecution

Ecological

Distribution

4 3

of 17 (Agkistrodon taylori) should be allocated to the CR

category, those with an EVS of 15 or 16 (Craugastor

mexicanus, Sceloporus parvus, Lampropeltis mexicana,

Salvadora bairdi, Thamnophis — pulchrilatus, _ T:

sumichrasti, Crotalus aquilus, C. intermedius, C.

polystictus, and C. triseriatus) to the EN category, and

those with an EVS of 13 or 14 (Lithobates montezumae,

L. spectabilis, Crocodylus moreletii, Barisia imbricata,

Gerrhonotus infernalis, Corytophanes hernandezii,

Sceloporus aeneus, §S. bicanthalis, S. minor, S.

mucronatus, Lepidophyma occulor, Conopsis biserialis,

C. lineata, Leptophis diplotropis, Masticophis schotti,

Pantherophis emoryi, Pituophis deppei, Trimorphodon

tau, Geophis mutitorques, G. semidoliatus, Rena dulcis,

R. myopica, Thamnophis scalaris, Metlapilcoatlus

nummifer, and Crotalus ravus) to the VU category. Thus,

these 31 species comprise 25.4% of the 122 LC species,

leaving 91 species that likely should remain in the LC

category, at least until more targeted surveys can be

undertaken.

March 2020 | Volume 14 | Number 1 | e224

Ramirez-Bautista et al.

Table 16. Components of the Environmental Vulnerability Scores (EVS) for members of the herpetofauna of Hidalgo, Mexico,

currently assigned to the [UCN Least Concern (LC) category. Non-native taxa are not included. * = country endemic.

Environmental Vulnerability Score (EVS)

Species Geographic Ecological Reproducuye

Wi tune Fa pay 1 Mode/Degree of

Distribution Distribution :

Persecution

—fa — le —fe ee Le ewe — —

—

eo art land lord mal (O71 — oe foe foes poe oe oe fo ol

oye] NTReIN]o N NInypols 10S) [Sed TN) Uo NO

Amphib. Reptile Conserv. 101 March 2020 | Volume 14 | Number 1 | e224

Herpetofauna of Hidalgo, Mexico

Table 16 (continued). Components of the Environmental Vulnerability Scores (EVS) for members of the herpetofauna of Hidalgo,

Mexico, currently assigned to the [UCN Least Concern (LC) category. Non-native taxa are not included. * = country endemic.

Environmental Vulnerability Score (EVS)

Species Geographic Ecological SED EOCHE LYE Total

Se ie eee tte Mode/Degree of

Distribution Distribution : Score

Persecution

| Conopsislineata* CT SC

| Conopsisnasus* ES

| Drymarchonmelanurus,

| Drymobius chloroticus, 8

| Ficimia streckeri* 8

| Lampropeltismexicana* ES 8S

| Leptophis diplotropis* TE SS 4

| Leptophismexicanus TE

| Masticophis flagellum CT 8

| Masticophis mentovarious, |

| Masticophis schoti CT 4S 4B

| Mastigodryas melanolomus |

| Pantherophisemoryi 4B

| Pituophiscatenifer CL A

| Pituophisdeppei* TS 4

| Pseudelaphe flavirufa,

| Salvadorabairdi* CS

| Salvadoragrahamiae_ CT

| Senticolistriaspis CE 8

| Tantillabocourti*

| Tantillarubra TS

| Trimorphodontau* SKB

| Adelphicos quadrivirgaum | A

| Amastridium sapperi__— CE

| Coniophanes imperialist

| Coniophanespiceivittis 88 T

| Diadophispunctatus,

| Geophis mutitorques* SB

| Geophis semidoliatus* SB

| Leptodeiramaculata 4

| Niniadiademata CT 8

| Pliocercuselapoides CT SO

| Rhadinaeadecorata, CT

| Rhadinaeahesperia

| Tropidodipsas sartorti_ TS

| Micrurusdiastema CS

| Micrurustener CT SS

|Renadulcis CT

| Renamyopica® TS TB

| Nerodiarhombifer CT SK

| Storeriadekayi CT CC KT

| Storeria storerioides* LS

| Thamnophis cyrtopsis

| Thamnophiseques CL

| Thamnophis marcianus

| Thamnophis proximus

| Thamnophis pulchrilatus* |

| Thamnophis scalaris* Sa

| Thamnophis sumichrasti* dE SS

| Scaphiodontophis annulaus | SS

| Crotalusmolossus CT Sas

| Crotalus polystictus* SS

| Crotalusravus* CS 4S

| Crotalus scutulatus aS

| Crotalustriseriatus* SS

| Metlapilcoatlus nummifer* | SSB

| Kinosternon hirtipes_ CE 8

| Kinosternon integrum* CE S88

Amphib. Reptile Conserv. 102 March 2020 | Volume 14 | Number 1 | e224

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Ramirez-Bautista et al.

Table 17. Number of herpetofaunal species in the four distributional status categories among the four physiographic regions of

Hidalgo, Mexico. Rank Order is determined by adding the numbers of Country Endemics and State Endemics.

Physiographic Region

Wey is Rank

Distributional Status Category

Non-endemics Country State Non-natives

Endemics Endemics

Sierra Madre Oriental

e e e

a eo

Trans-Mexican Volcanic Belt

Gulf Coastal Lowlands ae ae |

Mexican Plateau

Relative Herpetofaunal Priority

Johnson et al. (2015a) developed the concept of Relative

Herpetofaunal Priority (RHP), a simple metric used to

measure the relative importance of the herpetofaunal

species found in any geographic entity (e.g., a state or

physiographic region). Determining the RHP involves

the use of two methods: (1) calculation of the proportion

of state and country endemics as related to the entire

physiographic regional herpetofauna, and (2) computation

of the absolute number of high EVS category species in

each physiographic regional herpetofauna. The pertinent

data for these two methods are shown in Tables 17 and

18.

Based on the relative number of country and state

endemic species in each physiographic region and the

rank the regions occupy, the SMO region occupies rank

1, with 91 endemics out of a total of 166 species (54.8%,

Table 17). The other ranks are as follows: second = TMV

(63 of 84; 75.0%); third = MXP (51 of 79; 64.6%); and

fourth = GCL (31 of 93; 33.3%).

The data in Table 18 indicate that the rank ordering

of the four physiographic regions is the same as that

documented in Table 17. Based on the relative numbers

of high vulnerability species, the SMO region again

holds rank 1, with 47 high vulnerability species of a total

of 166 species (28.3%). The other ranks are as follows:

second = TMV (29 of 82; 35.4%); third = MXP (23 of 77;

29.9%); and fourth = GCL (15 of 91; 16.5%).

Based on the RHP analysis (Tables 17 and 18), the

most important physiographic region from a conservation

standpoint is clearly the SMO, because it harbors by far

the largest number of country and state endemics and

the greatest number of high vulnerability species. The

91 endemic species comprise 20 anurans (all country

endemics), 11 salamanders (eight country endemics

and three state endemics), 57 squamates (all country

endemics), and three turtles (all country endemics).

These 91 species are indicated in Table 4 with either

single or double asterisks. The SMO also contains 47

high vulnerability species, including seven anurans, nine

salamanders, 29 squamates, and two turtles. These 47

species and their respective EVS values are as follows:

Craugastor decoratus* (15)

Craugastor rhodopis* (14)

Eleutherodactylus longipes* (15)

Eleutherodactylus verrucipes* (16)

Bromeliohyla dendroscarta* (17)

Sarcohyla charadricola* (14)

Lithobates johni* (14)

Aquiloeurycea cephalica* (14)

Chiropterotriton arboreus* (18)

Chiropterotriton chondrostega* (17)

Chiropterotriton dimidiatus* (17)

Chiropterotriton mosaueri** (18)

Chiropterotriton multidentatus* (15)

Chiropterotriton terrestris* (18)

Isthmura gigantea* (16)

Pseudoeurycea leprosa* (16)

Abronia taeniata* (15)

Barisia imbricata* (14)

Sceloporus megalepidurus* (14)

Sceloporus minor* (14)

Sceloporus parvus* (15)

Lepidophyma occulor* (14)

Xenosaurus mendozai* (16)

Xenosaurus newmanorum®* (15)

Xenosaurus tzacualtipantecus* (16)

Lampropeltis mexicana* (15)

Pituophis deppei* (14)

Salvadora bairdi* (15)

Chersodromus rubriventris* (14)

Geophis latifrontalis* (14)

Geophis lorancai* (14)

Geophis turbidus* (15)

Hypsiglena tanzeri* (15)

Rhadinaea quinquelineata* (15)

Table 18. Number of herpetofaunal species in the three EVS categories among the four physiographic regions of Hidalgo, Mexico.

Rank Order is determined by the relative number of High EVS species. Non-native species are excluded.

j ? ; Medium : Rank

Sierra Madre Oriental

Trans-Mexican Volcanic Belt

Gulf Coastal Lowlands

Amphib. Reptile Conserv. 103 March 2020 | Volume 14 | Number 1 | e224

Herpetofauna of Hidalgo, Mexico

Thamnophis melanogaster* (15)

Thamnophis scalaris* (14)

Thamnophis sumichrasti* (15)

Agkistrodon taylori* (17)

Crotalus aquilus* (16)

Crotalus intermedius* (15)

Crotalus polystictus* (16)

Crotalus ravus* (14)

Crotalus totonacus* (17)

Crotalus triseriatus* (16)

Ophryacus smaragdinus* (14)

Terrapene mexicana* (19)

Kinosternon herrerai* (14)

Of these 47 species, 46 are country endemics and one 1s

a state endemic; their EVS values range from 14 to 19.

The TMV region occupies the second RHP rank,

with 63 country and state endemics (Table 17), including

12 anurans (all country endemics), 14 salamanders (10

country endemics and four state endemics), 36 squamates

(all country endemics), and one turtle (a state endemic;

Table 4). This region also harbors 29 high vulnerability

species (Table 18), including the following two anurans,

12 salamanders, and 15 squamates:

Eleutherodactylus longipes* (15)

Eleutherodactylus verrucipes* (16)

Aquiloeurycea cephalica* (14)

Chiropterotriton arboreus* (18)

Chiropterotriton chico** (18)

Chiropterotriton chiropterus* (16)

Chiropterotriton chondrostega* (17)

Chiropterotriton dimidiatus* (17)

Chiropterotriton magnipes* (16)

Chiropterotriton mosaueri** (18)

Chiropterotriton multidentatus* (15)

Chiropterotriton terrestris* (18)

Pseudoeurycea altamontana* (17)

Pseudoeurycea leprosa* (16)

Abronia taeniata* (15)

Barisia imbricata* (14)

Sceloporus megalepidurus* (14)

Sceloporus minor* (14)

Lampropeltis ruthveni* (16)

Pituophis deppei* (14)

Rhadinaea quinquelineata* (15)

Thamnophis pulchrilatus* (15)

Thamnophis scalaris* (14)

Thamnophis scaliger* (15)

Crotalus aquilus* (16)

Crotalus intermedius* (15)

Crotalus polystictus* (16)

Crotalus ravus* (14)

Crotalus triseriatus* (16)

Twenty-seven of these species are country endemics and

the other two are state endemics, and their EVS vary

from 14 to 18.

Amphib. Reptile Conserv.

The Mexican Plateau occupies rank three, with 51

country and state endemic species (Table 17), including 10

anurans (all country endemics), seven salamanders (five

country endemics and two state endemics), 33 squamates

(all country endemics), and one turtle (a country endemic;

Table 4). The region also contains 23 high vulnerability

species (Table 18), including the following one anuran,

five salamanders, and 17 squamates:

Sarcohyla charadricola* (14)

Chiropterotriton chiropterus* (16)

Chiropterotriton dimidiatus** (17)

Chiropterotriton mosaueri** (18)

Chiropterotriton multidentatus* (15)

Pseudoeurycea leprosa* (16)

Abronia taeniata* (15)

Barisia imbricata* (14)

Sceloporus megalepidurus* (14)

Sceloporus minor* (14)

Sceloporus parvus* (15)

Lampropeltis mexicana* (15)

Lampropeltis ruthveni* (16)

Pituophis deppei* (14)

Salvadora bairdi* (15)

Rhadinaea quinquelineata* (15)

Thamnophis melanogaster* (15)

Thamnophis scaliger* (15)

Thamnophis sumichrasti* (15)

Crotalus aquilus* (16)

Crotalus polystictus* (16)

Crotalus ravus* (14)

Crotalus triseriatus* (16)

Twenty-one of these species are country endemics and

the other two are state endemics, and their EVS vary

from 14 to 18.

The region occupying the fourth rank is the Gulf

Coastal Lowlands, which contains 31 country endemic

species (Table 17), including eight anurans (all country

endemics), one salamander (a country endemic), 22

squamates (all country endemics), and one turtle (a

country endemic; Table 4). This region also harbors

15 high vulnerability species (Table 18), including the

following five anurans, one salamander, seven squamates,

and two turtles:

Craugastor berkenbuschii* (14)

Craugastor decoratus* (15)

Craugastor rhodopis* (14)

Bromeliohyla dendroscarta* (17)

Lithobates johni* (14)

Bolitoglossa platydactyla* (15)

Lepidophyma occulor* (14)

Leptophis diplotropis* (14)

Chersodromus rubriventris* (14)

Rhadinaea quinquelineata* (15)

Thamnophis pulchrilatus* (15)

Agkistrodon taylori* (17)

March 2020 | Volume 14 | Number 1 | e224

Ramirez-Bautista et al.

. PNG Pleo

No. 33. Crotalus aquilus Klauber 1952. The D

is found “from the region of Lake Chapala, Jalisco, eastward

through Michoacan, Guanajuato, Querétaro, central San Luis

Potosi, and southeastward through northern Hidalgo and

northwestern Veracruz” (Lemos-Espinal and Dixon 2013:

249). This individual was encountered near Nopalillo, in the

municipality of Singuilican. Wilson (2013a) ascertained its EVS

as 16, placing it in the middle portion of the high vulnerability

category. It is allocated to the Least Concern category by the

IUCN, and is placed in the Special Protection (Pr) category by

SEMARNAT. Photo by Cristian Raul Olvera-Olvera.

- eG es SS

No. 35. Crotalus intermedius Troschel 1865. The Mexican

Small-headed Rattlesnake is distributed in “several disjunct

populations...in the central and southern highland region of

Mexico” (Campbell and Lamar 2004: 553). This individual

was found in El Encinal in the municipality of Singuilucan.

Wilson et al. (2013b) calculated its EVS as 15, placing it in

the lower portion of the high vulnerability category, the IUCN

has assessed it as Least Concern, and SEMARNAT listed this

rattlesnake as Threatened (A). Photo by Ferdinand Torres-

Angeles.

Amphib. Reptile Conserv.

Diamond-backed Rattlesnake is broadly distributed in the

United States and in Mexico. “In the United States, the

distribution...extends from Arkansas and _ north-central

Oklahoma westward to southeastern California and southward

through parts of Arizona, New Mexico, and much of Texas.

In Mexico, this species ranges from northeastern Baja

California through Sonora and northern Sinaloa, across most of

Chihuahua except for the Sierra Madre Occidental, throughout

Coahuila, Nuevo Leon, and Tamaulipas, and in the northeastern

parts of Durango and Zacatecas. It also occurs in Hidalgo and

Querétaro, and in parts of central and eastern San Luis Potosi,

as well as in extreme northern Veracruz” (Lemos-Espinal and

Dixon 2013: 250). This individual was found at Rancho Alegre,

in the municipality of San Agustin Metzquititlan. Wilson et al.

(2013a) calculated its EVS as 9, placing it at the upper limit of

the low vulnerability category. Its conservation status has been

evaluated as Least Concern by the IUCN, and it is allocated to

the Special Protection (Pr) category by SEMARNAT. Photo by

Cristian Raul Olvera-Olvera.

J ye Roa

No. 36. Crotalus ravus Cope 1865. The Mexican Pygmy

Rattlesnake is distributed in “temperate montane regions of

south-central Mexico” (Heimes 2016: 463). This individual was

found at Cerro Hihuingo in the municipality of Tepeapulco.

Wilson et al. (2013b) calculated its EVS as 14, placing it at

the lower limit of the high vulnerability category, the IUCN

has evaluated it as Least Concern, and SEMARNAT lists this

rattlesnake as Threatened (A). Photo by Christian Berriozabal-

Islas.

March 2020 | Volume 14 | Number 1 | e224

park guards; S = systems of

administrative services; R

facilities for visitors.

Table 19. Characteristics of Natural Protected Areas in Hidalgo, Mexico. Abbreviations for Facilities available are as follows: A

pathways; and V

Herpetofauna of Hidalgo, Mexico

Crotalus totonacus* (17)

Trachemys venusta (19)

Kinosternon herrerai* (14)

Herpetofaunal

survey

completed

Fourteen of these species are country endemics and the

other one is a non-endemic, and their EVS range from

14 to 19.

Fifty-eight members of the Hidalgo herpetofauna are

allocated to the high vulnerability category (Table 12),

thus the proportion of these species recorded in the four

physiographic regions are as follows: SMO (81.0%);

TMV (52.7%); MXP (41.8%); and GCL (27.3%).

These data should figure prominently in conservation

management plans for the state.

Management

plan available

Occupied by

landowners

Protected Areas in Hidalgo

Generally, the purpose of natural protected areas is

to allow for the continued functioning of ecosystem

services that are dependent on the interactions among the

components of the atmosphere, hydrosphere, lithosphere,

and biosphere. Thus, the protected areas that do the best

job of guarding ecosystem services are those left in a state

that is as close to a natural state as possible (Ervin 2003;

Gaston et al. 2008). Unfortunately, in a world overrun

by populations of the principal invasive species, Homo

Sapiens, areas can be maintained in a natural state only if

they are overseen by professional conservation managers

who are assigned to legally constituted protected areas.

In an effort to assess the state of Hidalgo’s protected

areas, a variety of data on these areas was assembled

(Table 19). The number of these protected areas in Hidalgo

is relatively small (five) compared to, for example, the 14

found in the adjacent state of Puebla (Woolrich-Pifia et

al. 2017). These five areas are all administered by the

Mexican federal government and include a biosphere

reserve, an Area of Protection of Natural Resources, and

three national parks (Table 19). The five areas range in

size from 99.5 to 96,042.9 ha. The total area is 164,160.8

ha or 1,641.6 km?, which is 7.9% of the area of the state

(20,813 km/?; http://cuentame.inegi.org.mx/monografias/

informacion/hgo/). They were established during the

period of 1936 to 2000.

The representation of these areas among the four

physiographic regions is skewed heavily toward the

Trans-Mexican Volcanic Belt; inasmuch as four of the

five areas are located in this region, while the other area

is found within the Gulf Coastal Lowlands. Thus, there 1s

no representation within the Mexican Plateau or the Sierra

Madre Oriental. This fact has major consequences for the

protection of the herpetofauna of Hidalgo, especially

since the Sierra Madre Oriental is shown above to be the

most significant region in Hidalgo for the herpetofauna,

due to the presence of high numbers of endemic and high

vulnerability species.

Considering the range of facilities available in these

protected areas, only two of them have a full range (as

Facilities

available

Gulf Coastal

Lowlands

Volcanic Belt

—

Trans-Mexican

Volcanic Belt

Transmexican

Volcanic Belt

Transmexican

Volcanic Belt

Transmexican

Jurisdiction

Mexican

Federal

Government

Mexican

Federal

Government

Mexican

Federal

Government

Mexican

Federal

Government

Mexican

Federal

Government

Municipalities

Acatlan, Atotonilco El

Grande, Eloxochitlan,

Huasca de Ocampo,

Meztitlan, San Agustin

Metzquititlan, Metepec,

Zacualtipan de Angeles,

El Cardonal

Acaxochitlan,

Cuautepec de Hinojosa

Mineral del Chico,

Mineral del Monte y

Pachuca de Soto

Pacula, Jacala de

Ledezma, Zimapan y

Nicolas Flores

96,042.9

42,129.4

Decree (dd/

mm/yyyy)

27-11-2000

20-10-1938

Category

Reserva de

la Biosfera

Barranca de

Metztitlan

Cuenca

Hidrografica

del Rio Necaxa

EI Chico pom 06-07-1982 2,739. 0

Amphib. Reptile Conserv. 106 March 2020 | Volume 14 | Number 1 | e224

Ramirez-Bautista et al.

Table 20. Distribution of herpetofaunal species in Natural Protected Areas of Hidalgo, Mexico, based on herpetofaunal surveys. *

= species endemic to Mexico; ** = species endemic to Hidalgo; and *** = non-native species.

Natural Protected Areas

Barranca de ‘ Cuenca Los

Metztitlan EnGSOEranca del Marmoles

Rio Necaxa

Anura(isspecies) |

Bufonidae species)

[incilius oceidemalis® TT

Jinciliwsvalliceps

[Rhinellahornibitis TT

[Craugastoridae species) |

Craugastorangusti

[Craugastorrhodopis* | TT

[Bleutherodactylidae(I species) [| t

Eleutherodactylus verrueipes* | ene ee

Hylidae (6 species) SSS a et

————<—

Eee

ithobarsspecubiisn | —*+d| id |

EC a

CSC

[Amerstomatonerapeciey | ——*i|—SSi

ES ec a

Ce ee

Ce OS

Eo a

Squamata Seapets) «| SST |

a SG

[Geronotwtocepnane | | |_| | + _

Amphib. Reptile Conserv. 107 March 2020 | Volume 14 | Number 1 | e224

Herpetofauna of Hidalgo, Mexico

Table 20 (continued). Distribution of herpetofaunal species in Natural Protected Areas of Hidalgo, Mexico, based on herpetofaunal

surveys. * = species endemic to Mexico; ** = species endemic to Hidalgo; and *** = non-native species.

Natural Protected Areas

Cuenca

Hidrografica del | El Chico

Rio Necaxa

Los

Marmoles

Barranca de

Metztitlan

[Gerrhonotus ophiurus® TT

[Phrynosomatidae(10species) ||

eS Se

[Seeloporusaeneus® |

Sra bei? |

[Scetoporus mucronams* | HT

[Seeloporuspanus® TE

a a

Sceloporus spinosus*

Sceloporus torquatus*

Sceloporus variabilis

——

Seinchine uwpesesy PC

Er | a a

Spnenomorphicne apace) | «iT —SSSCid SSC Si

Ee nn

Oe (EC

ESC nn

Fe | [T

Ce | Cs [

Bomenionndeme®

a A

a ES ES

ee FT CC A

Fe a

Fr ES

Ce A

iiptaimaanei® p+ fi id

Amphib. Reptile Conserv. 108 March 2020 | Volume 14 | Number 1 | e224

| : | |

Ramirez-Bautista et al.

Table 20 (continued). Distribution of herpetofaunal species in Natural Protected Areas of Hidalgo, Mexico, based on herpetofaunal

surveys. * = species endemic to Mexico; ** = species endemic to Hidalgo; and *** = non-native species.

Natural Protected Areas

Cuenca

Taxa Barranca de Los

Metztitlan

Hidrografica del | El Chico ; Tula

Marmoles

eodnaenmonat ere

————4 Necaxa

ie a Yo a

Etapidae(ispecies) | ET

pMicrurustener Ee

[Leptotyphlopidae(ispeciesy | FE

Renamvopica® Te

[Natricidae(W0species) |

[Nerodiarhombifer |

[Storeriahidaigoensis® |

[Storeriastorerioides® TET

[Thamnophiseyrtopsis |

Thamnophiseqes TT

[Thamnophis proximus |

[Thamnophispuchritatus® |

[Thamnophisscaaris® |

[Thamnophisscaiger® |

[Thamnophissumichrasi® | ET

[Viperidae(Sspeciesy |

Cromalusaguius® TE

Cromiusarox |

[Cromaiusmotossus |

[Cromatustriserias® |

Ophryacussmaragdinus® |

[Testudines (species) |

[Kinosternidae(Ispeciesy | FE

[Kinosternonimegram®™ | ET

[Torat(7sspeciesy | SF | Tt

indicated in Table 19). A major problem with all of these

five areas is that some amount of each is occupied by

landowners. Fortunately, however, management plans

are available for four of the five areas.

Herpetofaunal surveys have been completed for only

two of the five areas, so obviously surveys need to be

completed for the remaining three. Even though surveys

are incomplete for the protected areas in Hidalgo, the

available information on the distribution status of species

known to occur in each of these areas have been collated,

and are shown in Table 20 and summarized in Table 21.

Of the 200 native species that make up the

herpetofauna of Hidalgo, only 78 (39.2%) have been

recorded from the five areas combined. The numbers

of species recorded from these areas range from 13 in

Amphib. Reptile Conserv.

Parque Nacional Tula to 44 in Parque Nacional Los

Marmoles. Of the 103 country endemic species known

from Hidalgo, 51 (49.5%) are found in the five areas

combined. Of the 92 non-endemic species in the country,

only 25 (27.2%) are recorded for these areas. Two of the

four state endemic species (50.0%) are documented for

only two of the five areas (Parques Nacionales El Chico

and Los Marmoles). Finally, on the positive side, none

of the three non-native species has been recorded in

any of the protected areas. These data demonstrate that

completion of herpetofaunal surveys will be a major step

toward assessing the conservation needs of the Hidalgo

herpetofauna.

Of the 122 native species not found in any of the

five protected areas, 53 are country endemics, two are

March 2020 | Volume 14 | Number 1 | e224

Herpetofauna of Hidalgo, Mexico

Table 21. Summary of the distributional status of herpetofaunal species in Protected Areas in Hidalgo, Mexico. Total = total number

of species recorded in all of the listed protected areas.

Distributional status

Non-endemic Country State Endemic Non-native

(NE) Endemic (CE) (SE) (NN)

Number

Protected area of

species

RB Barranca de Metatitlan Ea

Necaxa

a —$—‘--t——

Fait a ee ee eee eee

Total tT

state endemics, and 67 are non-endemics. The 53 country

endemics not recorded in any of the protected areas are:

Incilius marmoreus

Craugastor berkenbuschii

Craugastor decoratus

Craugastor mexicanus

Eleutherodactylus longipes

Eleutherodactylus nitidus

Bromeliohyla dendroscarta

Sarcohyla arborescandens

Sarcohyla bistincta

Sarcohyla charadricola

Sarcohyla robertsorum

Lithobates johni

Lithobates montezumae

Bolitoglossa platydactyla

Chiropterotriton arboreus

Chiropterotriton chiropterus

Chiropterotriton magnipes

Isthmura gigantea

Pseudoeurycea leprosa

Norops naufragus

Anelytropsis papillosus

Sceloporus megalepidurus

Sceloporus scalaris

Scincella silvicola

Holcosus amphigrammus

Lepidophyma sylvaticum

Xenosaurus mendozai

Xenosaurus newmanorum

Xenosaurus tzacualtipantecus

Conopsis nasus

Ficimia olivacea

Lampropeltis mexicana

Lampropeltis polyzona

Lampropeltis ruthveni

Leptophis diplotropis

Tantilla bocourti

Chersodromus rubriventris

Geophis latifrontalis

Geophis lorancai

Geophis turbidus

Rhadinaea hesperia

Rhadinaea marcellae

Amphib. Reptile Conserv.

Rhadinaea quinquelineata

Epictia wynni

Thamnophis melanogaster

Agkistrodon taylori

Crotalus intermedius

Crotalus polystictus

Crotalus ravus

Crotalus totonacus

Metlapilcoatlus nummifer

Terrapene mexicana

Kinosternon herrerai

The two unrecorded state endemics are:

Chiropterotriton chico

Chiropterotriton terrestris

The 67 non-endemics are:

Anaxyrus punctatus

Incilius nebulifer

Eleutherodactylus cystignathoides

Eleutherodactylus guttilatus

Scinax staufferi

Smilisca baudinii

Tlalocohyla picta

Trachycephalus vermiculatus

Leptodactylus fragilis

Leptodactylus melanonotus

Hypopachus variolosus

Rhinophrynus dorsalis

Scaphiopus couchii

Notophthalmus meridionalis

Crocodylus moreletii

Basiliscus vittatus

Corytophanes hernandezii

Laemanctus serratus

Norops laeviventris

Norops lemurinus

Norops petersii

Norops sericeus

Coleonyx elegans

Ctenosaura acanthura

Sceloporus cyanogenys

Sceloporus serrifer

March 2020 | Volume 14 | Number 1 | e224

Ramirez-Bautista et al.

Rattlesnake occurs “from the Mohave Desert to northern

Sonora, and from extreme southern New Mexico and the Big

Bend region of Texas southward across the Mexican Plateau

to its southern edge” (Heimes 2016: 467). This individual

was found San José Atlan, in the municipality of Nopala.

Wilson et al. (2013b) calculated its EVS as 11, placing it in

the lower portion of the medium vulnerability category, the

IUCN assessed it as Least Concern, and SEMARNAT lists this

rattlesnake under the category of Special Protection (Pr). Photo

by Christian Berriozabal-Islas.

No. 39. Ophryacus smaragdinus Grinwald, Jones, Franz-

Chavez, and Ahumada-Carillo 2015. The Emerald Horned Viper

is known from “east-central Hidalgo, west-central Veracruz,

northeastern Puebla, and north-central Oaxaca” (Griinwald et

al. 2015: 398). This individual was located at Santa Catarina,

in the municipality of Tenango de Doria. Woolrich et al. (2017)

indicated its EVS to be 14, placing it at the lower limit of the

high vulnerability category. Its conservation status has not

been determined by the IUCN and this species is not listed by

SEMARNAT. Photo by Ferdinand Torres-Angeles.

Amphib. Reptile Conserv.

Dusky Rattlesnake is distributed in Aguascalientes, Ciudad de

México, Durango, Estado de México, Guanajuato, Guerrero,

Hidalgo, Michoacan, Morelos, Nayarit, Puebla, Querétaro, San

Luis Potosi, Tamaulipas, Tlaxcala, Veracruz, and Zacatecas

(Ramirez-Bautista et al. 2014). This individual was secured at

Los Reyes, in the municipality of Acaxochitlan. Wilson et al.

(2013a) estimated its EVS as 16, placing it in the middle portion

of the high vulnerability category. Its conservation status 1s

evaluated as Least Concern by the IUCN, but this species is not

listed by SEMARNAT. Photo by Ferdinand Torres-Angeles.

No. 40. Kinosternon herrerai (Stejneger, 1925). Herrera’s

Mud Turtle is distributed “in east-central Mexico, in southern

Tamaulipas, eastern San Luis Potosi, northern Veracruz,

Hidalgo, and Puebla” (Lemos-Espinal and Dixon 2013:

84). This individual was found at Laguna de Atezca, in the

municipality of Molango de Escamilla. Wilson et al. (2013a)

calculated its EVS as 14, placing it at the lower limit of the

high vulnerability category. Its conservation status has been

considered as Near Threatened by the IUCN, and is placed in

the Special Protection (Pr) category by SEMARNAT. Photo by

Christian Berriozabal-Islas.

March 2020 | Volume 14 | Number 1 | e224

Herpetofauna of Hidalgo, Mexico

Plestiodon tetragrammus

Lepidophyma flavimaculatum

Boa imperator

Coluber constrictor

Drymobius chloroticus

Drymobius margaritiferus

Ficimia streckeri

Lampropeltis annulata

Leptophis mexicanus

Masticophis flagellum

Masticophis mentovarius

Mastigodryas melanolomus

Oxybelis aeneus

Pituophis catenifer

Pseudoelaphe flavirufa

Salvadora grahamiae

Spilotes pullatus

Tantilla rubra

Adelphicos quadrivirgatum

Amastridium sapperi

Coniophanes imperialis

Coniophanes piceivittis

Hypsiglena jani

Imantodes cenchoa

Imantodes gemmistratus

Leptodeira maculata

Ninia diademata

Pliocercus elapoides

Rhadinaea decorata

Sibon nebulatus

Tropidodipsas sartorii

Micrurus diastema

Rena dulcis

Storeria dekayi

Thamnophis marcianus

Scaphiodontophis annulatus

Bothrops asper

Crotalus scutulatus

Trachemys venusta

Kinosternon hirtipes

Kinosternon scorpioides

Clearly, a major conservation goal regarding the

Hidalgo herpetofauna is to document the occurrence of

these 122 species, which comprise 61.0% of the native

herpetofauna, in one or more of the extant protected areas

in the state, as well as determining what other areas could

be designated 1n order to provide perpetual protection for

the entire herpetofauna. Based on the distributions noted

above, it is likely that such additional areas would need

to be established in the Sierra Madre Oriental and the

Mexican Plateau regions of the state.

Conclusions and Recommendations

Conclusions

A. Currently the herpetofauna of Hidalgo consists of

203 species, including 42 anurans, 17 salamanders, one

Amphib. Reptile Conserv.

crocodylian, 137 squamates (44 lizards and 92 snakes),

and six turtles.

B. The four physiographic regions recognized in Hidalgo

harbor from 77 species in the Mexican Plateau to 166 in

the Sierra Madre Oriental.

C. The number of species that are shared among

physiographic regions varies from 13 between the Trans-

Mexican Volcanic Belt and the Gulf Coastal Lowlands

to 72 between the Sierra Madre Oriental and the Gulf

Coastal Lowlands. The Coefficient of Biogeographic

Resemblance values range from 0.14 between the Trans-

Mexican Volcanic Belt and the Gulf Costal Lowlands to

0.65 between the Mexican Plateau and the Trans-Mexican

Volcanic Belt. The UPGMA dendrogram depicts two

distinct clusters; one between the Mexican Plateau and

Trans-Mexican Volcanic Belt (two montane regions),

and another between the mountainous Sierra Madre

Oriental and Gulf Coastal Lowlands. That the latter two

regions cluster is predicated on them sharing a significant

number of generalist species that typically occur on the

Atlantic/Gulf versant from the southern USA, through

Mexico and Central America, and into northern South

America.

D. The level of herpetofaunal endemism in Hidalgo is

relatively high. Of the 200 species making up the native

herpetofauna, 108 (54.0%) are endemic to either the

country of Mexico or the state of Hidalgo. Most of the

endemic species are country endemics (104 or 96.3%),

while only four are limited to the state of Hidalgo. All

four of the state endemics are plethodontid salamanders

of the genus Chiropterotriton.

E. The distribution status of the 203 species comprising

the Hidalgo herpetofauna is as follows (in order of

decreasing species numbers): country endemics (104;

51.2%); non-endemics (92; 45.3%); state endemics

(four; 2.0%); and non-natives (three; 1.5%).

F. The principal environmental threats are deforestation,

livestock, roads, pollution of water bodies, myths and

other cultural factors, and diseases.

G. The conservation status of the Hidalgo herpetofauna

was assessed using the SEMARNAT, IUCN, and EVS

systems. As in prior MCS papers, the SEMARNAT system

was found to be of minimal value, inasmuch as only 93

(46.5%) of the 200 native species have been evaluated

by this system. Of the 93 species presently assessed,

two are considered Endangered (P), 32 Threatened (A),

and 59 Special Protection (Pr). A comparison of the

SEMARNAT and distributional categorizations indicates

that one of the two Endangered species is a non-endemic

and the other is a country endemic species. Of the 32

Threatened species, nine are non-endemics and 23 are

country endemics. Of the 59 Special Protection species,

March 2020 | Volume 14 | Number 1 | e224

Ramirez-Bautista et al.

Mud Turtle ranges from “central Sonora to the Rio Verde in

Oaxaca, but it also is widespread throughout the central and

southern portion of the Mexican Plateau (Lemos-Espinal and

Dixon 2013: 86-87). This individual was found in the Valle

del Mezquital, in the municipality of Alfajayucan. Wilson et al.

(2013a) determined its EVS as 11, placing it in the lower portion

of the middle vulnerability category. Its conservation status has

been ascertained as Least Concern by the IUCN, and it is placed

in the Special Protection (Pr) category by SEMARNAT. Photo

by Christian Berriozabal-Islas.

21 are non-endemics, 36 are country endemics, and two

are state endemics.

H. The results of the IUCN assessment system (by

category and proportion) are: CR (seven of 200 species;

3.5%); EN (14; 7.0%); VU (14; 7.0%); NT (seven; 3.5%);

LC (122; 61.0%); DD (four; 2.0%); and NE (32; 16.0%).

I. In addition, application of the EVS system of

conservation assessment to the 200 native species of

Hidalgo, showed that the categorical values increase

from low vulnerability (66 species; 33.0%) to medium

(76; 38.0%), and then decrease to high vulnerability (58;

29.0%).

J. Comparing IUCN and EVS conservation status

categorizations to one another showed that 60.3% of

the EVS high vulnerability species are placed in one of

the three IUCN threatened categories (CR, EN, or VU),

and 54.1% of the low vulnerability species are in the LC

category. As noted previously for other areas, the results

of the application of these two conservation assessment

systems to the Hidalgo herpetofauna do not correspond

well with one another.

K. An examination of the conservation status of the

species placed in the [UCN DD, NE, and LC categories

demonstrates that many of these 158 species (79.0% of

the 200 native species) have been evaluated inadequately

compared to their respective EVS values, so we highly

recommend these species be reevaluated to better

indicate their prospects for survival.

L. The Relative Herpetofaunal Priority (RHP) measure

was used to ascertain the conservation significance of

Amphib. Reptile Conserv.

the four regional herpetofaunas in Hidalgo. This analysis

indicates that the most significant regional herpetofauna

is that of the Sierra Madre Oriental, as it contains the

largest numbers of endemic species and high vulnerability

species. The other three physiographic regions arranged

in decreasing order of significance based on numbers of

both endemic and high vulnerability species are: Trans-

Mexican Volcanic Belt; Mexican Plateau; and Gulf

Coastal Lowlands.

M. Only five protected areas are established in Hidalgo,

all administered by the federal government. Collectively,

the size of these areas comprises only 7.9% of the area of

the state. Four of these five areas are located in the Trans-

Mexican Volcanic Belt, which is only the second most

significant physiographic region in the state. None of the

areas 1S located in the Sierra Madre Oriental, which is by

far the most significant region, based on the numbers of

both endemic and high vulnerability species. Only two of

the five areas feature the full array of necessary facilities.

In addition, all five areas are occupied by landowners.

Management plans, however, are available for four of the

five areas. Herpetofaunal surveys are completed for only

two of the five areas.

N. Collated herpetofaunal records for each of the five

protected areas indicate that only 78 of the 203 species

occupying the state have been recorded from the five

areas combined. Of these 78 species, 51 are country

endemics, which is 49.0% of the total of 104 such

species in Hidalgo. Non-endemic species comprise 25

of 92 (27.2%) in the state. Only two of the four state

endemic species (50.0%) are recorded and only in one of

the protected areas. One good sign is that, to date, none

of the three non-native species recorded in Hidalgo are

known from any of the protected areas.

O. Future conservation efforts need to be directed

toward establishing the presence of the remaining 122

herpetofaunal species in the existing system of protected

areas, aS well as determining what other protected

areas could be developed in order to provide perpetual

protection for the entire herpetofauna of Hidalgo.

Presumably, such areas would need to be established in

the Sierra Madre Oriental and the Mexican Plateau.

Recommendations

A. Our purpose for writing this eleventh entry in

the Mexican Conservation Series is to document

the composition, physiographic distribution, and

conservation status of the 200 native species

comprising the herpetofauna of Hidalgo. In examining

the conservation status of these species, the EVS

methodology placed them into low, medium, and high

vulnerability categories in numbers which increased

from 66 (low) to 76 (medium) and then decreased to

March 2020 | Volume 14 | Number 1 | e224

Herpetofauna of Hidalgo, Mexico

58 (high). The Relative Herpetofaunal Priority measure

revealed that the most significant physiographic region

in Hidalgo for conservation is the Sierra Madre Oriental,

as it contains the most endemic species and species of

high vulnerability. Unfortunately, there are no protected

areas located within this region in Hidalgo, so the most

fundamental conservation challenge is to correct this

imbalance in the design of the protected area system in

Hidalgo.

B. The next most important conservation challenge is to

determine the presence in the existing protected areas

of the 122 herpetofaunal species that have not been

previously recorded in any of them and, beyond this

step, to ascertain which additional protected areas could

be established so that the entire native herpetofauna can

be protected in perpetuity. Likely, such additional areas

would need to be established in the Sierra Madre Oriental

and Mexican Plateau regions.

C. After the presence of the entire native herpetofauna

of Hidalgo in the protected areas system has been

ascertained, then the next step will be to establish

monitoring programs to guarantee the long-term survival

of these creatures.

D. Such steps need to be taken with the greatest dispatch,

as Hidalgo is the 17" most populous state in Mexico and

the eighth most densely populated.

“At this point in the fight to solve the climate crisis, there

are only three questions remaining: Must we change?

Can we change? Will we change?”

—AI] Gore (2017)

Acknowledgments.—We thank a variety of students

for their help with data collection, including Daniel

Lara-Tufifo, Luis M. Badillo-Saldafia, Diego Juarez-

Escamilla, Raquel Hernandez-Austria, Itzel Magno-

Benitez, Alejandro Ramirez-Pérez, Roberto Hernandez,

Victor Vite-Silva, Adrian lLeyte-Manrique, Aaron

Garcia-Rosales, Osiel Barrera, Carmen Serrano, Sean

Rovito, Cristian Raul Olvera-Olvera, Ferdinand Torres-

Angeles, Mirna G. Garcia-Castillo, Paola Lazcano-

Juarez, and Gustavo Rivas. We also thank three reviewers

for comments that greatly improved the manuscript.

We also are indebted to the authorities and residents of

the different municipalities of the state for providing

logistical support during the field work, and the curators

of the Herpetological Collection from Universidad

Autonoma del Estado de Hidalgo, National Collection

of Amphibians and Reptiles (CNAR) of the Institute of

Biology, and Amphibian and Reptile Collection of the

Museum of Zoology “Alfonso L. Herrera,” Faculty of

Sciences, both from the National Autonomous University

of Mexico (UNAM), for access to their collections. This

study was supported by projects Diversidad Bioldgica

Amphib. Reptile Conserv.

del Estado de Hidalgo FOMIX-CONACYT 43761,

FOMIX-HGO-2008-95828, FOMIX 2012/191908,

CONABIO JMO01, and Fomix-CONACyT-191908

Biodiversidad del Estado de Hidalgo-3a. Collecting

permits (SEMARNATO08-017-A, HESSX1304811,

SEMARNAT-SGPA/DGVS/02726/10 and SGPA/

DGVS/11746/13) were issued to ARB. We are also

indebted to Lia Berriozabel-Varela, Mirna G. Garcia-

Castillo, Aaron Garcia-Rosales, Raquel Hernandez-

Austria, Daniel Lara-Tufifio, Paola Lazcano-Juarez,

Cristian Raul Olvera-Olvera, Sean Rovito, and Ferdinand

Torres-Angeles, for the use of their photographic images.

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Aurelio Ramirez-Bautista began his herpetological research career as an undergraduate student

at the Los Tuxtlas Biological Field Station, Veracruz, Mexico. Aurelio received his Bachelor’s

in Biology from Universidad Veracruzana in Veracruz, Mexico, while his Master’s in Science

and doctorate were from the Universidad Nacional Autonoma de México (UNAM); and he

received a postdoctoral appointment at the University of Oklahoma, Norman, Oklahoma, USA.

His main research involves studies on ecology, demography, reproduction, conservation, and

life history evolution, using amphibians and reptiles of Mexico as models. Aurelio was president

of the Sociedad Herpetologica Mexicana and is currently Associate Editor of Mesoamerican

Herpetology. Aurelio was a professor at UNAM, and is currently a professor at Universidad

Autonoma del Estado de Hidalgo (UAEH) teaching population ecology, herpetology, and natural

history of amphibians and reptiles. He has authored or co-authored 295 peer-reviewed papers

and books on herpetology, ecology, life history evolution, sexual size dimorphism, reproduction,

global climate change, potential distribution, demography, conservation, behavior, and thermal

ecology. He has graduated 71 students (44 undergraduates, 18 Masters, and seven Ph.D. students);

and served as an external advisor for Ph.D. students at Brigham Young University, University

of Miami, and Eastern Carolina University, USA. Aurelio has received several national (Helia

Bravo Hollis Award by the Technical Council of Scientific Research of UNAM, member of the

National System of Researchers level II), and international awards (Donald Tinkle Award by

Southwestern Association of Naturalists), and he has the profile PRODEP (Programa para el

Desarrollo Profesional Docente) at UAEH.

Amphib. Reptile Conserv.

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Amphib. Reptile Conserv.

Ramirez-Bautista et al.

Uriel Hernandez-Salinas earned his Bachelor’s, Master’s and Ph.D. degrees at the Universidad

Autonoma del Estado de Hidalgo. Uriel is a herpetologist and co-author of three books:

Herpetofauna del Valle de México: Diversidad y Conservacion, Lista Anotada de los Anfibios

y Reptiles del Estado de Hidalgo, México, and Los Anfibios y Reptiles del Estado de Hidalgo:

Diversidad, Biogeografia y Conservacion. He is a full-time professor and curator-in-charge

of the scientific collection of amphibians and reptiles at CIIDIR Durango. Uriel has authored

or co-authored several peer-reviewed papers, and teaches Environmental Management II and

Fauna Management in the master’s and doctoral programs. In 2015, he became a member of

the National System of Researchers, level 1. His main topics of interest are biodiversity, species

richness, biogeography, and evolution of life histories of various species of the amphibians and

reptiles of Mexico.

Raciel Cruz-Elizalde is a Mexican herpetologist who received his B.Sc. in Biology, and M.Sc.

and Ph.D. in Biodiversity and Conservation, from the Universidad Autonoma del Estado de

Hidalgo (UAEH). Raciel is interested in the ecology, life history evolution, diversity, and

conservation of the amphibians and reptiles of Mexico. He has authored or co-authored several

publications including papers, notes, book chapters and books about the ecology, life history

evolution, sexual size dimorphism, reproduction, and conservation of amphibians and reptiles.

Raciel’s current research includes the life history evolution of diverse lizard species of genus

Sceloporus, conservation issues in natural protected areas, and the analysis of ecological and

morphological traits in the composition of amphibian and reptile assemblages.

Christian Berriozabal-Islas earned his bachelor’s degree and his master’s and Ph.D. degrees

in the Biodiversity and Conservation program, all at Universidad Aut6noma del Estado

de Hidalgo. Christian is a herpetologist interested in species diversity, thermal ecology,

functional diversity, climatic change, and distributional patterns using amphibians and

reptiles as biological models. Currently, he is a professor at the Universidad Autonoma del

Estado de Hidalgo. Christian has been involved in projects regarding environmental education

and wildlife conservation in rural communities, was a co-author of the book Los Anfibios y

Reptiles del Estado de Hidalgo, México: Diversidad, Biogeografia y Conservacion (2014),

and has authored or co-authored several papers on diversity, ecology, and climate change. He

has a great interest in the natural history of the turtles of Mexico.

Israel Moreno-Lara is a herpetologist who recently graduated from the Universidad

Autonoma del Estado de Hidalgo (UAEH) after completing a thesis project on the

conservation, protection, and trafficking of arboreal lizards of the genus Abronia (Anguidae)

under the mentorship of Aurelio Ramirez-Bautista and Raciel Cruz-Elizalde. Israel is working

on the conservation of herpetofauna through educational projects using identification cards

for the amphibian and reptile species of Hidalgo, and he is actively involved in scientific

dissemination campaigns at UAEH.

Dominic L. DeSantis is an Assistant Professor of Biology at Georgia College and State

University, Milledgeville, Georgia, USA, in the Department of Biological and Environmental

Sciences. Dominic’s research interests broadly include the behavioral ecology, conservation

biology, and natural history of herpetofauna. Much of his current research focusses on

integrating multiple longitudinal monitoring technologies to study the proximate and ultimate

drivers of spatial strategies and activity patterns in snakes. Dominic accompanied Vicente

Mata-Silva, Eli Garcia-Padilla, and Larry David Wilson on survey and collecting expeditions

to Oaxaca in 2015, 2016, and 2017, and is a co-author on numerous natural history publications

produced from those visits, along with an invited book chapter on the conservation outlook

for herpetofauna in the Sierra Madre del Sur of Oaxaca. Overall, Dominic has authored or

co-authored over 50 peer-reviewed scientific publications.

117 March 2020 | Volume 14 | Number 1 | e224

Amphib. Reptile Conserv.

Herpetofauna of Hidalgo, Mexico

Eli Garcia-Padilla is a herpetologist primarily focused on the ecology and natural history of the

Mexican herpetofauna, particularly the Mexican states of Baja California, Tamaulipas, Chiapas,

and Oaxaca. His first experience in the field was researching the ecology of the insular endemic

populations of the rattlesnakes in the Gulf of California, and his Bachelor’s degree 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 peer-reviewed scientific publications. Eli is currently the

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; http://

www.naturalista.mx). One of his main passions is environmental education, and for several years

he has been using audiovisual media to reach large audiences in promoting the importance of the

knowledge, protection, and conservation of Mexican biodiversity. Eli’s interests include wildlife

and conservation photography, and his art has been published in several recognized scientific,

artistic, and educational books, magazines, and websites. His present research project involves

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, USA,

and he has extensive experience studying the herpetofauna of Mesoamerica, especially southern

Mexico. Jerry is the Director of the 40,000-acre Indio Mountains Research Station, was a co-

editor of Conservation of Mesoamerican Amphibians and Reptiles and co-author of four of its

chapters. Jerry has authored or co-authored over 100 peer-reviewed papers and is the Mesoamerica/

Caribbean editor for the Geographic Distribution section of Herpetological Review. One species,

Tantilla johnsoni, has been named in his honor. Presently, he is an Associate Editor and Co-chair of

the Taxonomic Board for the journal Mesoamerican Herpetology.

Vicente Mata-Silva is a herpetologist originally from Rio Grande, Oaxaca, Mexico, whose 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, USA (UTEP). Vicente is an Assistant Professor of Biological

Sciences at UTEP in the Ecology and Evolutionary Biology Program, and Co-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.

Larry David Wilson is a herpetologist with extensive 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 (MLS. and Ph.D.

degrees). Larry has authored or co-authored more than 420 peer-reviewed papers and books on

herpetology, including 18 papers from 2013-2019 on the EVS measure and the MCS surveys of the

composition, distribution, and conservation status of the herpetofauna of different states in Mexico

and other regions in Central America. Larry is the senior editor of Conservation of Mesoamerican

Amphibians and Reptiles and a co-author of seven of its chapters. His other major books include

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 71 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. In 2005, he was designated a Distinguished Scholar in the Field of Herpetology at the Kendall

Campus of Miami-Dade College. Currently, Larry is a Co-chair of the Taxonomic Board for the

journal Mesoamerican Herpetology.

118 March 2020 | Volume 14 | Number 1 | e224

Official journal website:

amphibian-reptile-conservation.org

Amphibian & Reptile Conservation

14(1) [General Section]: 119-131 (e225).

First record of Raorchestes longchuanensis Yang and LI,

1978 (Anura: Rhacophoridae) from northeastern Bangladesh

suggests wide habitat tolerance

‘Hassan Al-Razi, ‘Marjan Maria, ‘Sabit Hasan, and **Sabir Bin Muzaffar

'Faculty of Life and Earth Science, Department of Zoology, Jagannath University, Dhaka, BANGLADESH *Department of Biology, United Arab

Emirates University, Al Ain, UNITED ARAB EMIRATES

Abstract.—Raorchestes is a genus of small bush frog characterized by an absence of vomerine teeth, direct

development without free swimming larvae, and a transparent gular pouch while calling. During a larger study

on canopy fauna in the northeastern region of Bangladesh, five specimens of a small bush frog were collected

from Satchari National Park in June and July 2017. This species was confirmed as Raorchestes longchuanensis

using both morphometric and genetic analyses. Although this species was originally described from Yunnan,

China, the authors speculated that it may be found in neighboring countries adjacent to the original records,

including northern Myanmar, Thailand, Laos, and Vietnam. However, the current finding suggests that the

species could be more widespread and resilient, spanning westwards through to northeastern India and

Bangladesh. Data are also provided on coloration, habitat, natural history, and vocalizations of this little-known

species. Although the species is designated Least Concern according to IUCN, more comprehensive studies

should be undertaken to better understand its biology and population status to aid in a more comprehensive

global conservation assessment.

Keywords. Amphibian, Asia, bush frog, DNA, range extension, Satchari National Park

Citation: Al-Razi H, Maria M, Hasan §S, Bin Muzaffar S. 2020. First record of Raorchestes longchuanensis Yang and Li, 1978 (Anura: Rhacophoridae)

from northeastern Bangladesh suggests wide habitat tolerance. Amphibian & Reptile Conservation 14(1): 119-131 (e225).

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: 15 February 2019; Accepted: 10 August 2019; Published: 23 March 2020

Introduction Asia have been classified variously into a range of tropical

moist, tropical evergreen, and several other forest types

Amphibians around the world are declining due to a based on the composition of vegetation (Champion and

variety of factors including habitat loss, competition Seth 1968).

with invasive species, emerging infectious diseases, and Recent work using phylogeographic approaches

climate change (Blaustein et al. 2010; Fisher et al. 2009; — suggests that South Asian and Indo-Pacific forest types

Gray et al. 2009; Hayes et al. 2010; Pounds et al. 2006; = exhibit more compositional overlap than previously

Whittaker et al. 2013). Globally, nearly one-third of — imagined (Slik et al. 2018). Incidentally, many of these

known amphibian species are threatened with extinction, areas are just beginning to be adequately explored for

and at least 42% of known amphibian species are amphibian biodiversity and many species of forest-

experiencing population losses (Whittaker et al. 2013). associated amphibians are being described (Biju and

These problems are acute in tropical Asia where large = Bossuyt 2009; Biju et al. 2010a,b, 2011, 2014; Hasan

swaths of forest and wetland areas have already been _ et al. 2014a; Mahony 2011; Mahony et al. 2013, 2018;

replaced with agricultural land or urban environments, §Purkayastha and Matsui 2012). Not surprisingly, many of

while existing patches are under constant pressure from — these newly discovered species are already under some

human expansion. Thus, forest biodiversity within the — form of threat and could be heading towards extinction

tropical and subtropical belts has been the focus of major (IUCN 2015). Bangladesh hosts at least 64 species of

efforts in biodiversity conservation worldwide due to anurans (Khan 2015), with several recently discovered

their recognized importance in climate stabilization, new species (Hasan et al. 2012, 2014a; Howlader 2011;

economic value (e.g., timber production or non-timber | Howlader et al. 2015, 2016) and new country records

forest products), and to a lesser extent ecotourism (Hasan et al. 2010; Khan 2001; Mahony and Reza

potential (Muzaffar et al. 2011). Tropical forests in South 2007a,b, 2008; Reza 2008), and several others are likely

Correspondence. * s_muzaffar@uaeu.ac.ae (ORCID: 0000-0001-9195-1677)

Amphib. Reptile Conserv. 119 March 2020 | Volume 14 | Number 1 | e224

Raorchestes longchuanensis in Bangladesh

to be added to the list in the near future (Khan 2015).

Raorchestes Biju, Shouche, Dubois, Dutta, and

Bossuyt (2010) is a genus of small-sized frogs with

direct development in the Rhacophoridae family. A total

of 62 Raorchestes species are known from India, Nepal,

Bangladesh, Myanmar, Thailand, Laos, Southern China,

Vietnam, Cambodia, and West Malaysia (Frost 2019).

Although four species are reported from Northeast India,

only two species of this genus are found in Bangladesh

(Frost 2019; IUCN 2015). Of the 62 Raorchestes species,

58 are from South Asia and only four species, including

Raorchestes longchuanensis (Yang and Li 1979), occur in

Southeast Asia and southern China (Frost 2019).

In this study, Raorchestes longchuanensis is reported

from the north-eastern region of Bangladesh, based on

observations made during a survey conducted as part

of a larger study on canopy fauna in this region. Forests

in the northeastern regions are considered to be tropical

moist deciduous or tropical semi-evergreen forests

(Champion and Seth 1968) and are located in the south

and southwest of the Indian states of Meghalaya, Assam,

and Nagaland, along with comparable forest types such as

tropical moist deciduous and subtropical broad-leaved hill

forests (Champion and Seth 1968). The objectives of this

study were (1) to diagnose the presence of this frog species

hitherto reported only from southern China; (11) to provide

morphometric, acoustic, and natural history data on this

species; and (iii) to provide comments on its biogeography.

Methods

Study Area

Surveys were conducted in Satchari National Park (SNP),

located in Habiganj District, Sylhet in northeastern

Bangladesh. The park encompasses an area of about 243

ha (Mukul et al. 2010) and is located on Raghunandan

hill, under Paikpara Union, Chunarughat Upazilla.

The topography of this area is undulating with slopes

and hillocks, ranging from 10—150 m in elevation. The

climate is mainly tropical with high rainfall (300-800

mm) concentrated during the monsoon from June to

September, with temperatures ranging from 25-32 °C. In

the remaining dry period of the year, the region experiences

occasional, low rainfall and lower temperatures of 12—25

°C (Mollah et al. 2004). The total annual average rainfall

is 4,162 mm (Mollah et al. 2004). A number of small,

sandy-bedded streams drain the forest during the rainy

season. The maximum and minimum temperatures of the

area are 32 °C and 12 °C, respectively, and the relative

humidity fluctuates between 74% and 90% (Mollah et al.

2004).

Field Survey

Five specimens were collected in June and July 2017.

Only acoustic searches were used to locate the calling

Amphib. Reptile Conserv.

ageregations (Heyer et al. 1994). The temperature,

humidity, habitat, perch height, and behavioral activities

were recorded after encounter. All individuals collected

were males since they were located based on their calls.

No female individuals were found. The location of

each collected specimen was recorded by a GPS device

(Garmin eTrex 10), and temperature and humidity were

recorded with a digital thermometer and hygrometer,

respectively. Video footage of a calling male individual

was also recorded using a DSLR camera (Nikon d7200

with 55—300 mm Vr lens). Five adult male individuals

were collected from two different field surveys, fixed in

formalin, and preserved in 70% ethanol. A small amount

of thigh muscle was collected (as described below)

from four specimens, and these tissue specimens were

preserved in 70% ethanol. Specimens were deposited in

the Museum of the Department of Zoology, Jagannath

University, Dhaka, Bangladesh (reference numbers:

JnUZool-A0117 to JnUZool-A0517).

Morphometrics

The following morphometric measurements of the

specimens were taken before preservation with a digital

caliper (to the nearest 0.1 mm): snout-vent length, from

tip of snout to vent (SVL); head length, distance between

tip of snout and rear of mandible (HL); head width, at

angle of jaw (HW); eye diameter, horizontal diameter of

eye (ED); tympanum diameter, maximum diameter of

tympanum (TD); eye-nostril distance, distance between

anterior canthus of eye and nostril (END); snout length,

from anterior canthus of eye to tip of snout (SL); inter

orbital distance, least distance between proximal edges of

upper eyelids (IOD); internarial distance, least distance

between nostrils (IND); upper eyelid width, maximum

transverse distance of upper eyelid measured from inner

edge to outer edge (UEW); eye-tympanum distance,

least distance between eye and tympanum (ETD); thigh

length, distance from middle of vent to knee (THL);

shank length, distance between knee and heel (SL); foot

length, from base of inner metatarsal tubercle to tip of Toe

IV (FL); hand length, from base of outer palmar tubercle

to tip of Finger [V (HAL); width of 1* to 4" finger disks,

measured at the widest point on the finger disk (FinDW),;

lengths of 1* to 4" fingers, from base of palm to tip of

respective finger (FinL, F-I to F-IV); lengths of 1° to

5" toes, from inner metatarsal tubercle region to tip of

respective toe (ToeL, T-I to T-V); and width of 1% to 5"

toe disks, greatest horizontal distance between edges of

Toe I disks (ToeDW).

DNA Extraction and Amplification

The procedure described in Vences et al. (2012) was

followed for DNA extraction. Approximately 1.5 mm? of

thigh muscle tissue was excised from each specimen for

extraction. The PCR amplification and sequencing of the

March 2020 | Volume 14 | Number 1 | e224

Al-Razi et al.

Fig 1. External features of Raorchestes longchuanensis. (A)

Calling position of an adult male individual where transparent

gular sack is prominent; (B) Ventral aspect of hand with

rounded, disked fingertips; (C) Ventral aspect of foot with

rounded disked, reddish toe tips.

16S rRNA gene were performed following Palumbi et al.

(1991) and Bossuyt et al. (2004), respectively. Primers

of 5’°-GCCTGTTTATCAAAAACAT-3’ (16Sar-L) and

5’-CCGGTCTGAACTCAGATCACGT-3’ (16Sbr-H)

as forward and reverse primers for 16S (Palumbi et al.

1991), were used for this study. PCR amplifications

were performed in a 20 ul reaction volume with the

following cycling conditions: an initial denaturing step

at 95 °C for 3 min; 40 cycles of denaturing at 95 °C

for 30 s, annealing at 50 °C for 30 s, and extending at

72 °C for 45 s; and a final extending step of 72 °C for

5 min. The amplified products were sent to Ist Base

Laboratories, Malaysia, for sequencing. The sequences

were checked manually using the program Chromas lite

2.01 (http://www.technelysium.com.au/chromas_lite.

html). The sequences have been submitted to GenBank

(Accession nos: MH699074, MN193412, MN193413,

and MN193414).

To infer the phylogenetic position of the current

species, homologous sequences for 16S rRNA genes of

24 species were downloaded from the NCBI GenBank

database (Table 1). Sequences were aligned using the

MUSCLE tool in MEGA X (Kumar et al. 2018), and

alignments were checked visually and corrected manually

when necessary. Alignment gaps were treated as missing

data. The best fit model for nucleotide substitution was

selected from 24 models available in MEGA X (Kumar

et al. 2018) based on the minimum Akaike Information

Criterion (AIC) value (Posada and Crandall 2001).

Maximum Likelihood was performed in MEGA X. The

general time reversal nucleotide substitution model with

Amphib. Reptile Conserv.

gamma distribution with Invariant Sites (GTR+G+I),

obtained as a best fit model in the model test (AIC =

3932.32, InL = -1903.84), was used for constructing

the phylogenetic tree based on the maximum likelihood

method by using 1,000 bootstrap replicates in MEGA X

(Kumar et al. 2018). Interspecific and intraspecific mean

uncorrected pairwise distances were computed in MEGA

X (Kumar et al. 2018).

Call Recording and Analysis

The call of a single male individual (JnUZool- A0317)

was recorded with a DSLR camera (Nikon d7200 with

55-300 mm VR lens) on 2 August 2017. The sound

was extracted from the video clip by using Windows

Movie Maker 2012 software. The microphone was

approximately 1—-1.5 m away from the calling male.

Air temperature and humidity were taken with a digital

hygrometer. Call analysis used Raven Pro Ver. 1.5 (Charif

et al. 2010; Bioacoustics Research Program 2011).

Measured parameters were call-group duration, inter-call

group interval, duration of intervals between pulses, call

duration, and call rate of a call bout comprised of nine

call groups.

Results

First Observation

During the surveys, this species was observed for the

first time on 5 June 2017. The day was cloudy and after

a rainfall before dusk (1810 h to 1840 h) the surveyors

heard several tik-tik-tik-tik calls from the bamboo bushes,

not very far from the narrow road of the SNP visitor’s

watch tower. The bamboo bushes were entered and the

search for the frogs commenced. As there was still day

light, the search for the frog was conducted without any

light. After more than 20 min an adult male individual

was found on a bamboo tree. The male was calling

continuously while keeping its anterior part angled

downwards (Fig. 1A). The position of the frog was 1.5 m

above the ground. The individual was observed for a few

minutes, then collected as a sample specimen.

Species Identification

The species was initially identified as Raorchestes

longchuanensis based on the limited morphological

characters described by Yang and Li (1978) in their

original description of the species. Raorchestes

longchuanensis 1s a small species in which adult males

are 22.9 + 0.9 mm (n = 5), one-fourth of the toes are

webbed, and the disk of the third finger is smaller than

the tympanum diameter. This specimen was confirmed

as R. longchuanensis using the BLAST technique with

a sequence similarity of more than 99% (492 of 496

bp). Identification was also confirmed with Maximum

March 2020 | Volume 14 | Number 1 | e224

Raorchestes longchuanensis in Bangladesh

Table 1. 16S rRNA sequences for Raorchestes species and Kurixalus eiffingeri from GenBank, with accession numbers and

localities. SNP = Satchari National Park.

Species GenBank accession no. Locality Source

01 R. longchuanensis MH699074 SNP, Bangladesh This study

02 R. longchuanensis MN193412 SNP, Bangladesh This study

03 R. longchuanensis MN193413 SNP, Bangladesh This study

04 R. longchuanensis MN193414 SNP, Bangladesh This study

05 R. longchuanensis GQ285675 China Yu et al. 2010

06 R. ghatei KF366385 Western Ghats, India Padhye et al. 2013

07 R. gryllus AB933309 Vietnam Nguyen et al. 2014

08 R. shillongensis MG980282 Meghalaya, India Unpublished

09 R. shillongensis MG980283 Meghalaya, India Unpublished

10 R. parvulus MH590204 Gunung Jerai, Malaysia Chan et al. 2018

11 R. menglaensis EU924621 Yunnan, China Yu et al. 2009

12 R. tuberohumerus KP137388 Western Ghats, India Padhye et al. 2015

13 R. bombayensis KF767502 Castlerock village, Karnataka, India Padhye et al. 2013

14 R. bombayensis EU450019 Castlerock village, Karnataka, India Biju and Bossuyt 2009

15 R. indigo KM596557 Western Ghats, India Vijayakumar et al. 2014

16 R. ponmudi EU450026 Ponmudi, Western Ghats, India Biju and Bossuyt 2009

17 R. ponmudi KM596576 Ponmudi, Western Ghats, India Vijayakumar et al. 2014

18 R. aureus KM596540 Western Ghats, India Vijayakumar et al. 2014

19 R. montanus KM596552 Western Ghats, India Vijayakumar et al. 2014

20 R. signatus KM596561 Western Ghats, India Vijayakumar et al. 2014

21 R. marki JX092719 Western Ghats, India Vijayakumar et al. 2014

27 R. charius KU169985 Karnataka, India Buu et al. 2016

23 R. beddomii EU449998 Western Ghats, India Biju and Bossuyt 2009

24 R. chalazodes KJ619641 Western Ghats, India Das 2015

25 R. dubois JX092668 Koadaikanal, India Vijayakumar et al. 2014

26 R. munnarensis JX092655 Western Ghats, India Vijayakumar et al. 2014

27 R. chromasynchysi KM596543 Western Ghats, India Vijayakumar et al. 2014

28 Kurixalus eiffingeri DQ468673

likelihood phylogenetic analyses (Fig. 2) and the intra-

and interspecific genetic p-distances (0.2—0.3% and 5.0—

13.9% average values, respectively, Table 3).

Morphometrics

Snout-vent lengths of the five adult males ranged from

21.4—23.9 mm. Head width was larger than the length

of head (HW = 7.8-9.1 mm, HL = 6.30—7.17 mm). The

snout was pointed in the dorsal view; snout length was

larger than the eye diameter (SL = 2.90-3.12 mm, ED

= 2.50—2.86 mm); nostrils were oval and closer to snout

tip than to eye; vomerine teeth were absent; eyes were

small with a horizontal pupil. Fingers were web-less and

finger tips had disks that were rounded. Relative lengths

of fingers were as follows: I < II < IV < III (Table 2).

The thigh was slightly longer than the tibia (TL = 11.20-

12.10 mm, THL = 11.70-12.50 mm); Toe V was longer

than Toe III; relative lengths of toes were as follows: I <

Il < Ill < V < IV (Table 2). Subarticular tubercles were

prominent and rounded; outer metatarsal tubercles were

absent while inner metatarsal tubercles were present; and

toe tips possessed distinct, rounded disks (Fig. 1B—C).

Amphib. Reptile Conserv.

Okinawa Islands, Japan

Wuet al. 2016

Coloration

Considerable color variation was recorded in this species,

including four different body colors in the specimens that

could change in relation to the environment. A distinct

or faint “)(’ mark was always present on the dorsal

surface of the body (Fig. 3). Sometimes a narrow yellow

mid-dorsal line was present on the dorsal side (in one

of five specimens). Vocal sac was external, single, and

transparent with white spotting. The discs of the fingers

and toes were reddish, orange, or whitish. There was

a pale or prominent brown or black line between the

upper eyelids. The dorsum was of a dark chocolate, dark

brown, or pale brown coloration in the live specimens.

The dorsal coloration and the pale flanks became gray or

light brown to dark brown after preservation. Eyelids and

tympanic fold were blackish or brown (Fig. 3).

Habitat, Natural History, and Vocalization

The specimens of Raorchestes longchuanensis were

found in bushes inside the forest, and sometimes near

road sides or near human habitations. Usually they were

122 March 2020 | Volume 14 | Number 1 | e224

Al-Razi et al.

15 14]

Raorchestes longchuanensis (GQ285675)

Raorchestes longchuanensis (MH699074)

Raorchestes longchuanensis (MN193412) |

Raorchestes longchuanensis (MN193413)

Raorchestes longchuanensis (MN193414

Raorchestes gryllus (AB933309)

100 , Raorchestes shillongensis (MG980282)

Raorchestes shillongensis (MG980283)

Raorchestes parvulus (MH590204)

33“ Raorchestes menglaensis (EU924621)

70 Raorchestes tuberohumerus (KP137388)

Raorchestes bombayensis (KF767502)

33 ' Raorchestes bombayensis (EU450019)

Raorchestes ghatei (KF366385)

Raorchestes indigo (KM596557)

16 Raorchestes ponmudi (EU450026)

33 ' Raorchestes ponmudi (KM596576)

Raorchestes aureus (KM596540)

66 Raorchestes montanus (KM596552)

64+ Raorchestes signatus (KM596561)

? Raorchestes marki (Jx092719)

Raorchestes chanus (KU 169985)

Raorchestes beddomii (EU449998)

Raorchestes chalazodes (KJ619641)

39 Raorchestes duboisi (JX092668)

83 Raorchestes munnarensis (JX092655)

Raorchestes chromasynchysi (KM596543)

—————_—“+

0.050

Kunxalus eiffingen (0Q468673)

Fig 2. Maximum likelihood phylogenetic tree based on 16S rRNA genes, showing the identity of the specimens as Raorchestes

longchuanensis. The red box shows the specimens from the present study.

found living amongst the bamboo bushes and Calamus

palm bushes (Calamus spp.). Individuals usually were

perched on leaves and branches of small trees or bamboo

bushes. All individuals were found less than 2 m above

the ground. Rarochestes longchuanensis became active

with the onset of rain in the month of April. Breeding

males started calling from dusk and continued until 2400

h. During rain and in cloudy environments they became

active in the day time.

Advertisement calls occurred in call groups (Fig. 4).

The duration of the analyzed call was 20 s. The number

of call groups within this call was nine, and the number

of pulses varied from three to six. The duration of each

call group was 0.526 + 0.142 s (n= 9) and it varied with

Amphib. Reptile Conserv.

the number of calls. The interval between call groups was

1.831 + 0.403 s (n= 8). Pulse duration was 0.013-—0.024

s (0.018 + 0.003 s, n = 42 pulses), and the duration of

intervals between pulses was 0.102—0.163 s (0.124 +

0.013 s, n = 33 intervals). The lowest frequency was

1.35 kHz and the highest frequency was 4.07 kHz. The

overall peak frequency of the calls was 2.76 kHz. The air

temperature was 27.4 °C at the time of the call.

Discussion

Several species of frogs have been recorded for the

first time or described from Bangladesh in recent years

(Asmat 2007; Hasan et al. 2012, 2014a,b; Howlader

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Raorchestes longchuanensis in Bangladesh

Table 2. Morphologic measurements (mm) of Raorchestes longchuanensis specimens collected from Satchari National Park,

Bangladesh (n = five males).

Characters Abbreviation JnUZool-A0117 = JnUZool- A0217 JnUZool-A0317 JnUZool- A017 JnUZool-A0418

Snout-vent length SVL 22.60 23.90 21.40 23.89 22:52

Head length HL 6.80 Talk 6.40 7.03 6.80

Head width HW 8.80 9.10 8.69 9.00 8.00

Eye diameter ED 2.75 2.86 2.50 27 2.66

Tympanum diameter TD 1.50 1.56 1.43 1.6 1.49

Eye-nostril distance EN 215 Dodo 1.88 2.20 1.96

Snout length SL 3.12 3.10 2.90 3.07 3.03

Interorbital distance IOD 3.18 3,52 3.28 3.40 3.40

Internarial distance IND 233 2.50 225 2.47 2.34

Upper eyelid width UEW E32 1.61 1.25 1.60 1.28

cite = ETD 0.72 0.82 0.70 0.75 0.71

Thigh length THL 12.34 12.50 11.70 12.41 12.26

Tibia length TL 12.00 12.10 11.20 12.08 12.00

Foot length FL 8.90 9.56 8.83 9.53 9.03

Hand length HAL Sie 6.21 5.69 6.17 6.06

Forearm length FLL 5.8 6.42 5.61 6.33 6.00

Finger I disk width Finl DW 0.50 0.56 0.47 0.56 0.48

Finger II disk width Fin2DW 0.80 0.85 0.79 0.87 0.80

Finger III disk width Fin3DW 1.30 1.47 1.26 1.56 1.31

Finger IV disk width Fin4DW 1.23 1.25 1.18 1.25 1.22

Finger I length FinlL 1.61 1.93 1.6 192 1.59

Finger IT length Fin2L 2.20 2.38 1.92 2.33 2.17

Finger III length Fin3L 4.14 4.36 4.04 43] 4.15

Finger IV length Fin4h 2.50 202 2.35 2:53 2.47

Toe I length ToelL 3,24 3.25 2.87 a2 3.20

Toe II length Toe2L 4.51 4.56 4,52 4.62 4.50

Toe III length Toe3L 6.96 6.99 6.93 7.08 6.95

Toe IV length Toe4L 8.71 8.98 8.69 9.04 8.68

Toe V length ToeS5L 6.35 6.08 6.77 6.96 6.30

Toe I disk width Toel DW 0.52 0.58 0.43 0.53 0.50

Toe II disk width Toe2DW 0.80 0.77 0.74 0.82 0.72

Toe III disk width Toe3 DW 0.88 0.87 0.82 0.90 0.85

Toe IV disk width Toe4DW 1.18 1.26 1.11 1.30 1.21

Toe V disk width ToeS5DW 0.86 0.98 0.76 1.02 0.87

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Al-Razi et al.

i a

ie t i

Fig 3. Color variation in Raorchestes longchuanensis. (A) Pale brown dorsum with faint “)(’ mark; (B) Dark brown dorsum with

prominent “)(” mark; (C) Dark chocolate color dorsum without mid-dorsal line; (D) Dark chocolate color dorsal side with narrow

yellow mid-dorsal line.

2011; Howlader et al. 2015, 2016; Khan 2015; Mahony

and Reza 2007a,b, 2008; Reza 2008; Mahony et al.

2009). This is not surprising since Bangladesh is a small

country situated on the boundary of two major ecological

realms, with the Indomalayan realm encompassing most

of Bangladesh and the beginning of the Palearctic realm

to the north (Udvardy 1975). The position of Bangladesh

on the edge of these biogeographic zones makes it prone

to hosting biota from both realms. These areas have

been also recognized as part of the Indo-Burma global

biodiversity hotspot, with significant abundance of

threatened and regionally endemic amphibians (Sechrest

et al. 2002; Jenkins et al. 2013). Until recently, only 49

species of amphibians were officially recorded from

Bangladesh (Hasan et al. 2014b). The recent rise in new

amphibian records constitutes an almost 65% increase

Amphib. Reptile Conserv.

in the anurans of Bangladesh since the late 1990s,

suggesting that exploration of the region’s amphibians is

just beginning (calculated from IUCN 2015; Khan 2015).

The tree frogs of Bangladesh have been particularly

under-studied. One species in the genus Philautus was

originally recorded from Bangladesh (Hasan and Feeroz

2014). This genus was originally considered to be

distinguishable from others by the absence of vomerine

teeth and its occurrence in South and Southeast Asia

(Bossuyt and Dubois 2001). However, further study

showed that Philautus exhibits diverse morphological

traits and a wide variety of life history strategies

(Bossuyt and Dubois 2001). Philautus from the Western

Ghats could be separated from Pseudophylautus (Sri

Lankan sister clade) and appeared to be distinct from

those in northeastern India. This led to the erection of a

March 2020 | Volume 14 | Number 1 | e224

Raorchestes longchuanensis in Bangladesh

Amplitude (kU)

Nw & oO ~ W

Frequency (KHz)

0.00

Amplitude (kU)

]

Amplitude (kU)

0 0.2 0.4 0.6 0.8 1 0

Time (s)

-10

-20

2 4 6 8 10 12 14 16 18 20

Time (Ss)

10 12 14 16 18 20

Time (s)

5 10 15 20 25

Time (ms)

Fig 4. Advertisement call of Raorchestes longchuanensis at ambient air temperature 27.4 °C. (A) A call consists of five call groups,

numbers on the top indicate pulse numbers in the respective call group. (B) Spectrogram of the call. (C) First call group with four

pulses. (D) A pulse of the 5" call group.

new genus Raorchestes (based on molecular phylogeny),

consisting of small frogs with no vomerine teeth, direct

development without free swimming larvae, and a

transparent gular pouch while calling (Biju et al. 2010).

At least 62 Raorchestes species have been named to

date, but many of these are species complexes and further

study could reveal cryptic species (Frost 2019). Many of

the Philautus species from across the distribution of this

genus have become subsumed into Raorchestes (Ghose

and Bhuiyan 2012; [UCN 2015). This includes the former

P. annandalii (Darjeeling Bush Frog) being changed to R.

annandalii, also recorded from the northeastern regions

of Bangladesh (IUCN 2015; Khan 2015, 2018). The

second species, R. parvulus (Karin Bubble-nest Frog)

reported by Mahony et al. (2009) and later by Ghose and

Bhutyan (2012), is considered to be a possible member

of a species group found in southeastern Bangladesh

(Mahony and Reza 2007a,b; Mahony et al. 2009; Khan

2015; IUCN 2016).

Raorchestes —longchuanensis

was __ previously

Amphib. Reptile Conserv.

126

considered endemic to southern China, restricted to some

prefectures in Yunnan province at 1,150—1,600 m above

sea level, adjacent to northern and northeastern Myanmar

(Yang et al. 2004). However, this species has already

been recorded from Vietnam (Orlov et al. 2002). Yang

et al. (2004) have suggested that the species was likely

to occur in neighboring countries including northern

Myanmar, Thailand, and Laos. The species has escaped

detection in some areas possibly due to its cryptic nature

and the general absence of major exploration in these

regions.

The findings reported here also point to the broader

floral similarities between Myanmar and northeastern

India (including states such as Tripura, Mizoram,

Manipur, and Nagaland) [Slik et al. 2018], which forms

a belt connecting our study site, and northern Myanmar

across into Yunnan, China. This study presents a range

extension of this species of approximately more than

600 km in the western direction from its type locality

(Fig. 5). An earlier study showed a range extension of

March 2020 | Volume 14 | Number 1 | e224

Al-Razi et al.

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March 2020 | Volume 14 | Number 1 | e224

127

Amphib. Reptile Conserv.

Raorchestes longchuanensis in Bangladesh

80°0’0.0”N

is]

te)

20°0'0.0"S

0 100 200 300 400km

/—+—_+—_+—

20°0'0.0”N

(0°0'0.0’W

8 120°0’0.0"W

160°0’0.0”W

Location

* New record

Countries

[1 Bangladesh

G8 Bhutan

8 China

[9 India

(9 Lao PDR

HS) Myanmar

M9 Nepal

HS Thailand

GS Vietnam

160°0’0.0"E

Fig 5. Map showing the previously reported and present records of Raorchestes longchuanensis.

more than 600 km in the east for this species (Orlov et

al. 2002). Thus, we suspect that the species has a much

wider distribution, and further studies in these regions

will confirm its occurrence. We hope that more studies,

especially those targeting arboreal species, will continue

in Bangladesh and in neighboring regions to document

and potentially save these species from the risks posed

by the perpetual exploitation of timber in these regions.

A comprehensive effort needs to be made to at least

safeguard the remaining wilderness areas in Bangladesh

and in neighboring regions.

Conclusions

This study adds Raorchestes longchuanensis as a new

species to the anuran fauna of Bangladesh. Although

currently assessed as Least Concern by the IUCN (Yang

et al. 2004), its geographic distribution and current

conservation status need to be re-evaluated, since its

habitats have further degraded since the last assessment.

The breeding biology, ecology, and habitat characteristics

of the species need to be determined to ensure long-term

conservation of this species in Bangladesh and elsewhere.

Acknowledgements.—We thank the Forest Department

of Bangladesh for their support and facilitation of this

work. The authors are thankful to Professor Dr. Abdul

Amphib. Reptile Conserv.

Alim, Chairman, Department of Zoology, Jagannath

University, for his support. We are thankful to Md.

Anisur Rahman (beat officer) and Mahbub Alam (range

officer) of Satchari National Park for their support during

the study. We are especially thankful to Tanvir Ahmed for

his support during field work.

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Hassan Al-Razi has a B.Sc. in Zoology and an M.Sc. in Wildlife and Biodiversity Conservation

from the Department of Zoology, Jagannath University, Dhaka, Bangladesh. Currently, Hassan is a

researcher at the same university, and he has been working on nocturnal animals for more than four

years. His research fields include the diversity, taxonomy, and breeding biology of herpetofauna,

as well as primate and small carnivore conservation.

Marjan Maria is a B.Sc. student in the Department of Zoology, Jagannath University, Dhaka,

Bangladesh. Marjan is a wildlife researcher interested in the diversity, taxonomy, and ecology of

herpetofauna, as well as nocturnal mammals and birds.

Sabit Hasan is an M.Sc. student in the Department of Zoology, Jagannath University, Dhaka,

Bangladesh. Sabit is currently involved in the study of various nocturnal animals. He is interested

in the ecology and conservation of wildlife.

Sabir Bin Muzaffar is a Professor in the Department of Biology, United Arab Emirates University.

Sabir is an ecologist with broad interests in animal movement, migration, community structure,

March 2020 | Volume 14 | Number 1 | e224

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Amphibian & Reptile Conservation

14(1) [General Section]: 132-139 (e226).

Geographic range extension for the Lobatse Hinge-back

Tortoise, Kinixys lobatsiana (Power, 1927), with first records

from the Soutpansberg region

'*Flora Ihlow, 7Ryan Van Huyssteen, ‘Melita Vamberger, 7Dawn Cory-Toussaint,

4Margaretha D. Hofmeyr, and ‘Uwe Fritz

'Museum of Zoology, Senckenberg Dresden, A.B. Meyer Building, 01109 Dresden, GERMANY *Soutpansberg Centre for Biodiversity and

Conservation, Medike Nature Reserve, Soutpansberg, SOUTH AFRICA *University of Venda Limpopo, SOUTH AFRICA *Chelonian Biodiversity

and Conservation, Department of Biodiversity and Conservation Biology, University of the Western Cape, Bellville 7535, SOUTH AFRICA

(deceased)

Abstract.—The Lobatse Hinge-back Tortoise, Kinixys lobatsiana (Power, 1927), has a small distribution range

in northern South Africa and adjacent Botswana. Local populations have been fragmented by degradation

and destruction of suitable habitat, resulting in this species being listed as Vulnerable by IUCN. Here, the

geographic distribution of K. lobatsiana is updated and several hitherto unpublished occurrences are reported,

which extend its distribution range to the north.

Keywords. Africa, chelonians, geographic distribution, Reptilia, South Africa, Testudinidae

Citation: Ihlow F, Van Huyssteen R, Vamberger M, Cory-Toussaint D, Hofmeyr MD, Fritz U. 2020. Geographic range extension for the Lobatse Hinge-

back Tortoise, Kinixys lobatsiana (Power, 1927), with first records from the Soutpansberg region. Amphibian & Reptile Conservation 14(1): 132-139

(e226).

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 January 2020; Accepted: 11 March 2020; Published: 25 March 2020

The Lobatse Hinge-back Tortoise, Kinixys lobatsiana

(Power, 1927), is near-endemic to northern South Africa.

The putative distribution range of this species covers an

area of approximately 93,000 km? (extent of occurrence),

which mainly falls within the South African province of

Limpopo but also extends into the adjacent North West,

Gauteng, and Mpumalanga provinces as well as into

neighboring south-eastern Botswana (Fig. 1; Hofmeyr

and Boycott 2018; Power 1927; TTWG 2017).

In the north, the distribution range was reported to

reach to south of the Soutpansberg mountain range

(Boycott 2014; Hofmeyr and Boycott 2018), with the sole

specimen recorded further north assumed to represent a

translocated individual (Boycott 2014; Broadley 1993;

Hofmeyr and Boycott 2018; TTWG 2017). The Lobatse

Hinge-back Tortoise was described as being closely

associated with rocky hillsides and outcrops (Boycott

2014; Boycott and Bourquin 2000; Broadley 1989;

Hofmeyr and Boycott 2018) and uses rock crevices and

abandoned animal burrows as hiding places (Bonin et

al. 2006; Broadley 1989). However, in the frame of the

present study the species was also found in open savanna

habitats, with only few rocks or burrows, using low shrubs

(Grewia sp. and Boscia albitrunca) for shelter (Fig. 2).

In some of these sites many living tortoises examined

had extensive burn marks on their shells, indicating that

fires represent a significant threat. While this habitat

type is naturally prone to burning, intentional fire as

a management tool (e.g., set to stimulate new growth

of nutritious vegetation) is often practiced on a higher

frequency than natural fires would occur. Thus, higher

abundances in rocky habitats might result from a higher

survival rate due to the availability of sufficient shelter

rather than a real habitat preference.

The species is considered to be the most arid-adapted

Kinixys (Branch 2008) and was reported from various

vegetation types across the central bushveld bioregion.

These include dense, short bushveld and thornveld, open

tree savannas, Burkea savannas, mixed thornveld, and

Combretum woodlands (Boycott 2014; Branch 2008;

Broadley 1989; Hofmeyr and Boycott 2018). Kinixys

lobatsiana was reported to be absent from highveld

grassland and subtropical lowveld (Boycott 2014;

Branch 2008; Broadley 1989; Hofmeyr and Boycott

2018). However, we also found the species in a mosaic of

highveld grassland and Loskop mountain bushveld in the

Ezemvelo Nature Reserve. Degradation and destruction

of suitable habitat have been identified as the main

Correspondence. *flora.ihlow@senckenberg.de, melita.vamberger@senckenberg.de, uwe.fritz@senckenberg.de, ryanvanhuyssteen@gmail.

com, nycteris.cory2saint@gmail.com

Amphib. Reptile Conserv.

March 2020 | Volume 14 | Number 1 | e226

lhlow et al.

|_| Putative distribution range

100 km

e ¢€

Polokwane

@ Mokopane

% o® @ @ re)

@O if ata O

Johannesburg

Fig. 1. Putative range of the Lobatse Hinge-back Tortoise (Kinixys lobatsiana) according to TTWG (2017), with historic records compiled

from scientific collections. Inset: K. /obatsiana from the Lapalala Wilderness Reserve. Photo by Flora Ihlow.

threats for K. lobatsiana. While land use varies, habitat

transformation is considered particularly severe in north-

eastern South Africa and Botswana, where approximately

20-25% of suitable habitat has been destroyed within

the last four decades, and a loss totalling 35-40% is

expected within the next 30-40 years (Hofmeyr and

Boycott 2018). Intact habitat appears to be fragmented

and is largely restricted to protected areas and private

reserves. Consequently, the IUCN conservation status

of K. /obatsiana has recently been elevated from Least

Concern to Vulnerable (Hofmeyr and Boycott 2018).

Here, the geographic distribution range of the species

is discussed based on genetically verified records, and

several occurrences are reported that considerably extend

its known range. Field research was conducted across most

of the South African distribution range of K. /obatsiana

as well as at selected sites containing suitable habitat

outside of its known range. Study sites were selected by

geo-referencing collection sites of museum specimens

mentioned by Broadley (1989, 1993) and other published

records (TTWG 2017). The dataset was supplemented

with selected observations (i.e., those where photographic

vouchers allowed species identification) from the

ReptileMap database (FitzPatrick Institute of African

Ornithology 2019; http://vmus.adu.org.za). Searches for

K. lobatsiana were carried out during the species’ daily

activity times and suitable weather conditions. GPS

coordinates were collected for each individual. All tortoises

were identified based on established morphological traits

and coloration patterns (Broadley 1993), measured, and

photographed from several angles.

Amphib. Reptile Conserv.

In addition, a small (0.2 ml) blood sample was

drawn from the subcarapacial sinus using a 1 ml syringe

and a 25-G needle for genetic verification of species

identification. Blood samples were either preserved using

Whatman paper (GE Healthcare, Munich, Germany) or

in analytical ethanol, and subsequently processed at the

molecular genetic laboratory of the Museum of Zoology,

Senckenberg Dresden, Germany. Species identification

was genetically verified using mtDNA _ fragments

containing the cyt b gene or the partial ND4 gene plus

adjacent DNA coding for tRNAs following methods

outlined in Kindler et al. (2012). European Nucleotide

Archive (ENA) accession numbers of voucher sequences

for extralimital records are listed in Table 1.

The following hitherto unreported occurrences extend

the range of the species in two regions to the north,

namely in the northern Waterberg region and the western

Soutpansberg area (Fig. 2). Sampling sites inthe Waterberg

region include the Lapalala Wilderness Reserve, covered

by a vegetation type classified as Waterberg mountain

bushveld (Dayaram et al. 2017), and the private Kudu

Canyon Reserve (central sandy bushveld; Dayaram et al.

2017) located in the vicinity of the Mokolo Dam Nature

Reserve. Sites in the western Soutpansberg area comprise

Sigurwana and the neighboring Goro Game Reserve

(Soutpansberg mountain bushveld; Dayaram et al. 2017;

Table 1). The Leshiba Wilderness and the Medike Nature

Reserve neighboring Sigurwana in the east provide

similar high-altitude habitat, but harbor K. spekii instead

of K. lobatsiana. However, additional searches targeting

high elevation habitat in the Medike Nature Reserve will

133 March 2020 | Volume 14 | Number 1 | e226

Kinixys lobatsiana range extension in South Africa

[4@@ kK, lobatsiana

4@@ K. spekii

e Putatively introduced

ie) Putative distribution range

100 km

= @

A e

“f Pretoria

Johannesburg

Polokwane

Mokopane

Fig. 2. Distribution range of the Lobatse Hinge-back Tortoise (Kinixys lobatsiana) according to TTWG (2017), with genetically confirmed

range extensions (dots), not yet processed samples (dots with bold black outline), and recent observations (triangles). Observations

marked with an asterisk refer to collection material in the Ditsong National Museum of Natural History (TM 36366, TM 67909, TM

79431). Right: Characteristic habitats from different parts of the distribution range. Photos by Flora Ihlow.

be required to fully clarify the distributions of K. spekii

and K. lobatsiana in this area. The samples reported here,

published records, and museum specimens consistently

suggest that K. spekii also inhabits the surrounding

lowlands (Schmidt 2002).

Only a few herpetological surveys have been

previously conducted within the Soutpansberg Mountain

Range (e.g., SARCA surveys contributing to Bates et al.

2014; Jacobsen 1989; Kirchhof et al. 2010). Most studies

either targeted areas covered with habitat unsuitable

for K. lobatsiana or focused on taxa with considerably

different activity profiles. As a result, K. /obatsiana has

not been recorded from the area thus far, except for a

single record from north of the Soutpansberg mountains

that was considered to refer to a misplaced individual

(Broadley 1993; Hofmeyr and Boycott 2018; TTWG

Table 1. Records extending the distribution range of the Lobatse Hinge-back Tortoise (Kinixys lobatsiana) to the north. Coordinates

obscured to protect populations against poaching. ENA= European Nucleotide Archive; MTD T = Museum of Zoology, Senckenberg

Dresden (Tissue Collection).

MTDT ENA number Study site Latitude Longitude Elevation

17056 LR746292 Lapalala Wilderness Reserve -23.8 28.3 1,160 m

17062 LR746293 Lapalala Wilderness Reserve -23.8 28.3 1,160 m

17064 LR746294 Lapalala Wilderness Reserve -23.8 28.3 1,100 m

17067 LR746295 Lapalala Wilderness Reserve -23.8 28.3 1,040 m

17071 LR746296 Lapalala Wilderness Reserve -23.7 28.3 1,100 m

17076 LR746297 Lapalala Wilderness Reserve -23.8 252 1,060 m

17078 LR746298 Lapalala Wilderness Reserve -23.9 28.3 1,160 m

17079 LR746299 Lapalala Wilderness Reserve -23.9 28.3 1,140m

20164 LR746300 Kudu Canyon -24.0 Deel 1,009 m

20169 LR746301 Kudu Canyon -24.0 27.7 930 m

20176 LR746302 Goro Game Reserve -22.9 29.4 1,086 m

20188 LR746303 Sigurwana -22.9 29.4 1,380 m

20189 LR746304 Sigurwana -22.9 29.4 1,360 m

20738 LR746305 Sigurwana -22.9 29.4 1,380 m

20739 LR746306 Sigurwana -22.9 29.4 1,440 m

Amphib. Reptile Conserv. 134 March 2020 | Volume 14 | Number 1 | e226

lhlow et al.

Fig. 3. Lobatse Hinge-back Tortoise (Kinixys lobatsiana, TM 36366) collected at Rochdale Farm, Waterpoort, from the Ditsong National

Museum of Natural History. Photo by Adriaan Jordaan.

2017). This record likely corresponds to a museum

specimen housed at the Ditsong National Museum of

Natural History (TM 36366) collected in 1964 by G.

Newlands on Rochdale Farm, Waterpoort. Rochdale

Farm lies on the northern slope of the Soutpansberg,

just east of Waterpoort. A morphological examination

of TM 36366 confirms that it represents K. /obatsiana

(Fig. 3). Broadley (1993) suggested that this specimen

might have been “swept through the gorge by the Sand

River during a flood” to reach its collection site, while

Boycott (2014) added translocation by humans as another

possible explanation. Considering that K. /obatsiana

is confirmed from two other properties on the northern

slope of the Soutpansberg Mountains, 22 km away from

the collection site of TM 36366, that record no longer

appears to be an outlier. The present evidence rather

suggests a continuous distribution of K. lobatsiana along

the northern slope of the western Soutpansberg.

Schmidt et al. (2005) reported K. spekii from the

Blouberg Nature Reserve. Unfortunately, this finding

could not be re-examined because neither access to the

reserve nor photographs of the tortoise could be obtained

so far. However, a specimen in the Ditsong National

Museum of Natural History (TM 67909) collected

from a farm located approximately 6 km west of the

Blouberg Nature Reserve (The Glen, Quarter Degree

Square 2328Bb) morphologically resembles K. spekii.

In addition, two photo vouchers (163852 and 167289),

uploaded to the virtual museum database (http://vmus.

adu.org.za) showing the same individual from the

southern lowlands of the Blouberg Nature Reserve, also

resemble K. spekii. However, another specimen from

Amphib. Reptile Conserv.

the Ditsong National Museum of Natural History (TM

79431) collected at Bochum, Limpopo (QDS 2329AC),

morphologically resembles K. /obatsiana. Thus, further

research is needed to clarify the distributions of these two

species in this area.

The present report shows that the distributions of K.

lobatsiana and K. spekii are still incompletely known.

This contradicts Branch et al. (1995), who concluded that

the distribution of South African tortoises and terrapins

is relatively well-documented. Misidentifications of

these two morphologically challenging species and the

application of an outdated taxonomy compromise the

previous assessments of their distributions. Kinixys

lobatsiana and K. spekii were lumped together with

other species under “K. belliana” before Broadley (1989,

1993) separated these taxa.

While Broadley (1993) stated that for K. lobatsiana

no evidence of sympatry with any other species exists,

later authors considered K. /obatsiana to co-occur with K.

spekii throughout its range (TTWG 2017). However, only

a few areas (Waterberg region, western Soutpansberg)

were found where both species occur in close proximity.

Juveniles and old specimens of both species tend to lack

the characteristic shell shape and color patterns used to

distinguish between the two species (Broadley 1993),

which leads to high misidentification rates, compromises

range estimates, and has implications for conservation

measures that are based on erroneous records and range

estimates (Figs. 4—5; Ihlow et al. 2019).

The most reliable morphological trait to distinguish

challenging adult specimens of K. spekii and K. lobatsiana

is the serrated posterior carapace rim in the latter species

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Kinixys lobatsiana range extension in South Africa

Fig. 4. Top: Juvenile Lobatse Hinge-back Tortoise (Kinixys lobatsiana, SCL = 81 mm) from Sigurwana, western Soutpansberg, Limpopo.

Bottom: Young Speke’s Hinge-back Tortoise (K. spekii, SCL = 106 mm) from Leshiba Wilderness, western Soutpansberg. Scale bar: 1

cm. Species identification of both tortoises was genetically confirmed. Photos by Flora Ihlow.

(Ihlow et al. 2019). However, this character does not

always suffice in juveniles (Fig. 4). To enable later re-

examination, photographs should document the dorsal,

ventral, and lateral aspects of each tortoise. In addition,

molecular genetic confirmation is recommended.

Branch et al. (1995) concluded that most South

African chelonian species are adequately protected

by existing reserves. However, he stressed that K.

lobatsiana was only recorded in three major reserves,

namely the Loskop Dam Nature Reserve (Mpumalanga),

the Ohrigstad Dam Nature Reserve (Mpumalanga), and

the Nylsvley Nature Reserve (Limpopo). The species

could be confirmed from a photo voucher taken by

an employee of the Loskop Dam Nature Reserve in

2017, but records from the other two reserves have to

Amphib. Reptile Conserv.

be considered uncertain. Only a single possible record

exists for the wider Ohrigstad area (Ditsong National

Museum of Natural History, TM 21329, a specimen

closely resembling K. /obatsiana, collected by P. van

Tonder in 1944). However, the Ohrigstad Dam Nature

Reserve itself is located well outside of the putative

range of the species. In the Nylsvley Nature Reserve, “K.

b. belliana” was reported from a single photographed

tortoise (Jacobsen 1977), which was identified as K.

lobatsiana by Broadley (1993), but Jacobsen (2008)

referred to the same record later as “K. belliana spekei

[sic]”, suggesting that K. spekii, and not K. lobatsiana,

occurs in the Nylsvley Nature Reserve. Unfortunately,

no hinge-back tortoise has been recorded there for many

years by the local rangers (pers. comm.) nor are there any

March 2020 | Volume 14 | Number 1 | e226

lhlow et al.

Fig. 5. Speke’s Hinge-back Tortoise (Kin

ixys spekii) from the vicinity of Vaalwater (left) and Lobatse Hinge-back Tortoise (K. /obatsiana)

from the Lapalala Wilderness Reserve (right) with very similar color patterns. Species identification of both tortoises was genetically

confirmed. Photos by Flora Lhlow.

photographs of hinge-back tortoises from this reserve

in any of the databases that were queried. Branch et al.

(1995) further highlighted the Marakele National Park

(former Kransberg National Park) and the Blyde River

Canyon Nature Reserve as important statutory protected

areas where the species has been recorded. However, we

are not aware of any contemporary records from either of

these reserves. In addition to these published localities,

the occurrence of K. Jobatsiana from the following sites

was confirmed:

Gauteng: Ezemvelo Nature Reserve (Bronkhorst-

spruit), Irene, Kalkheuwel, Leopard Lodge (Thiane

Wildlife Sanctuary), Monateng Safari Lodge (East

Lynne).

Limpopo: Bushfellows Lodge (Marble Hall), Goro

Game Reserve (Western Soutpansberg), Inkwe

Private Nature Reserve (Waterberg), Kalkfontein

Farm (Groblersdal), Kudu Canyon Farm (Lephalale),

Lapalala Wilderness Reserve (Waterberg), Lépellé

Lodge (Burgersfort), Sigurwana Lodge (Western

Soutpansberg), Thandabantu Game Lodge (Roos-

senekal), Welgevonden Nature Reserve (Waterberg).

Mpumalanga: Loskop Dam Nature Reserve.

North West: Kgaswane Nature Reserve (Rustenburg),

Koster (Rustenburg), land owned by the Bahurutshe

Ba Ga Lencoe Traditional Council (Moshana village),

Riekersdam Farm (Ramotshere Moiloa), Tswenyane

Safaris (Zeerust), Vaalkop Dam Nature Reserve

(Rustenburg).

While the species appears to be rare in most areas,

larger populations were recorded in the Waterberg area

(Lapalala Wilderness Reserve, the Welgevonden Nature

Reserve, and in the vicinity of the Mokolo Dam Nature

Reserve), in the vicinity of Kalkheuwel, and in the

vicinity of Lobatse, Botswana.

Acknowledgements.—We thank Arnaud Le Reux

and Henning Pienaar (Bela Bela); Jade Vanneste

(Bushfellows Game Lodge); Mark Bing, Liezille

Draper (Ezemvelo Nature Reserve); Dave Dewsnap

Amphib. Reptile Conserv.

(Goro Game Reserve); Debbie Van den Berg and Pieter

Ernst Kruger (Groenfontein Game Lodge); Courtney

Hundermark; Claudia and Arne Hofmann (Inkwe Private

Nature Reserve); Karina, Henk and Hawee Koelman

(Kalkfontein Farm); Jenny Hill, Allen Liebenberg, and

Tyron Clark (Kalkheuwel); Bernard Tabane and Thomas

Maramba (Kgaswane Mountain Reserve); Richard

Burrough, Britta Zawada, Jeremiah Ndlovu, and Piet

Lourens (Krokodilspruit); Anna Margaretha and Hendrik

Jacobus Nel (Kromellenboog Farm); Sandy and Anton

van Niekerk (Kudu Canyon); Annemieke and Hermann

Muller (Lapalala Wilderness Reserve); Barnie Fouché

(Lépélle Lodge); Kathryn and Peter Straughan (Leshiba

Wilderness); Hannes Botha and Jannie Coetzee (Loskop

Dam); Oldrich van Schalkwyk (Medike Nature Reserve,

Endangered Wildlife Trust); Kathrin Monaghan; Allison

Sharp and Terrence Anderson (Monateng Safari Lodge);

Kgosi Edwin Ikalafeng Lencoe and the Bahurutshe

Ba Ga Lencoe Traditional Council (Moshana village);

Karel Gronum Loots (Riekersdam Farm); Julia Poschel,

Anke Muller, and Anja Rauh (Senckenberg Dresden);

Liesel and Niel Wright as well as Daniel Stephanus

Booyens (Sigurwana); Sonja Carougo (Thandabantu

Lodge); Jeremy Thompson; Burger Koekemoer

(Tswenyane Safaris); the management of the Vaalkop

Dam Nature Reserve, Carmen Warmenhove, and Wade

Kilian (Welgevonden Game Reserve); Quintin Kruger

(Wolwefontein, Leopard Lodge, Bulgerivier); Leon

Fourié (Wonderboomhoek); and John Zoran for assistance

during field or laboratory work and for permitting research

on their properties. We are grateful to Anders G.J. Rhodin

for sharing an unpublished dataset of occurrence records

compiled for the latest TTWG checklist. We thank

Lauretta Mahlangu, Lemmy Mashinini, and Adriaan

Jordaan (Ditsong National Museum of Natural History)

for permission to examine specimens and for providing

photographs. Genetic investigations were conducted

in the Molecular Laboratory of Senckenberg Dresden

(SGN-SNSD-Mol-Lab). Ethical clearance was received

from the University of the Western Cape (AR 19/4/1).

Research was performed under national and provincial

collection permits: Limpopo ZA/LP/91608, North West:

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Kinixys lobatsiana range extension in South Africa

NW6124/10/2018, and Gauteng: CPF6-0210. Field

research was partly funded through the Mapula Trust

awarded to Margaretha D. Hofmeyr. Flora Ihlow profited

from a Margarethe Koenig Scholarship of the Zoological

Research Museum Alexander Koenig and is currently

supported by the German Science Foundation (DFG IH

133/1-1).

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lhlow et al.

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 predominantly focused

on the ecology, systematics, and distribution of Southeast Asian chelonians. She has published

numerous scientific papers on the herpetofauna of Southeast Asia. After her graduation from the

Rheinische Friedrich Wilhelms University (Bonn, Germany), Flora joined the phylogeography

department of Senckenberg, Dresden in 2017 to work on the systematics and distribution of

chelonians from southern Africa within the frame of a post-doctoral position. Flora is a member of

the IUCN/SSC Tortoise and Freshwater Turtle Specialist Group (TFTSG).

Ryan Van Huyssteen is a South African field biologist interested in reptile distribution, biogeography,

and ecology. Ryan currently lives in the Soutpansberg, Limpopo, South Africa, where he has been

working on reptiles and conservation for the past five years.

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 capsica 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.

Dawn Cory-Toussaint is currently a final year Ph.D. student at the University of Venda (South

Africa) studying the ecological impacts of opencast diamond mining on bats and the role that bats

can play as biological indicators in Limpopo. Dawn has a broad understanding and an unlimited

interest in the natural world. She has been involved in projects on diverse organisms ranging from

Killer Whales (satellite tagging and biopsy sampling) to beetles (Darkling Beetles of the Bushveld).

Being involved in projects that are broader than her current study, particularly in northern Limpopo,

is invaluable to her role in the management and conservation in the area where she currently resides

and works.

Margaretha D. Hofmeyr was Professor Emeritus at the Biodiversity and Conservation Biology

Department, University of the Western Cape, South Africa. She was 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 published extensively on the ecology and

phylogeography of sub-Saharan tortoises and turtles, and she was closely involved in conservation

projects on threatened tortoises. This work resulted in her being awarded the 2015 Sabin Turtle

Conservation Prize. She was a member, and Regional Vice-Chair for Africa, of the IUCN/SSC

TFTSG and coordinated the 2014 and 2018 Red List Assessment for South African tortoises and

freshwater turtles.

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 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 proceedings and

books, among them the two turtle volumes of the Handbook of Amphibians and Reptiles of Europe.

139 March 2020 | Volume 14 | Number 1 | e226

Official journal website:

amphibian-reptile-conservation.org

Amphibian & Reptile Conservation

14(1) [General Section]: 140-146 (e227).

urn:lsid:zoobank.org:pub: D6FE38AE-C4AF -40BE-AD69-93F197FB91E1

A new species of Petracola (Squamata: Gymnophthalmidae)

from Rio Abiseo National Park, San Martin, Peru

‘*Lily O. Rodriguez and 2?4Luis Mamani

'Museo de Historia Natural, Universidad Nacional Mayor de San Marcos (MUSM), Lima, PERU *Programa de Magister en Ciencias con mencion

en Zoologia, Departamento de Zoologia, Facultad de Ciencias Naturales y Oceanogrdaficas, Universidad de Concepcion, Barrio Universitario S/N,

Casilla 160C, Concepcién, CHILE *Museo de Historia Natural de la Universidad Nacional de San Antonio Abad del Cusco (MHNC), Plaza de

Armas s/n (Paraninfo Universitario), Cusco, PERU ‘4Museo de Biodiversidad del Pert (MUBI), Urbanizacion Mariscal Gamarra A—61, Zona 2,

Cusco, PERU

Abstract—A new species of Petracola is described from Rio Abiseo National Park in northeastern Peru,

where it inhabits grasslands above montane forest at 3,230 m asl. The new species is diagnosed by a unique

combination of morphometric, scalation, and color pattern characteristics, increasing the number of described

Petracola species to five.

Keywords. Andean lizard, Cercosaurini, Gran Pajatén, Huallaga basin, montane forest, Reptilia

Citation: Rodriguez LO, Mamani L. 2020. A new species of Petracola (Squamata: Gymnophthalmidae) from Rio Abiseo National Park, San Martin,

Peru. Amphibian & Reptile Conservation 14(1) [General Section]: 140-146 (e227).

[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 September 2019; Accepted: 23 February 2020; Published: 28 March 2020

Introduction

The genus Petracola Doan and Castoe 2005 is a lineage

of semifossorial lizards distributed in the central and

northern Andes of Peru (Doan and Castoe 2005; Eche-

varria and Venegas 2015; Kizirian et al. 2008; Kohler and

Lehr 2004; Uzzell 1970). These small lizards occur in

cloud forest, montane forest, and wet puna from 1,889 m

to 3,600 m asl (Echevarria and Venegas 2015; Kizirian et

al. 2008; Kohler and Lehr 2004).

The genus Petracola was created to resolve the poly-

phyly of Proctoporus (Doan and Castoe 2005) and was

nested in Cercosaurinae as sister lineage of a lineage

formed by Cercosaura, Dendrosauridion, Potamites,

Proctoporus, Selvasaura, and Wilsonosaura (Torres-Car-

vajal et al. 2016; Moravec et al. 2018; Vasquez-Restrepo

et al. 2019). It currently includes four species: P. angus-

tisoma Echevarria and Venegas 2015, P. waka Kizirian,

Bayefsky-Anand, Eriksson, Le, and Donnelly 2008, P.

labioocularis (Kohler and Lehr 2004), and P. ventrimac-

ulatus (Boulenger 1900). Compared to its close relatives,

P. ventrimaculatus has a relatively extensive geographic

distribution, including both sides of the Marafion River

(Kizirian et al. 2008); furthermore, osteological, mor-

phological, and ecological evidence suggest it could be

a complex of species (Echevarria 2014).

Correspondence. */rodriguez@cima.org.pe

Amphib. Reptile Conserv.

In 1987, during a faunal inventory expedition to the

upper part of the Rio Abiseo National Park, Peru, the first

author (LOR) collected five specimens with characteris-

tics matching the Petracola ventrimaculatus group and

deposited them in the herpetology collection of Museo

de Historia Natural de la Universidad Nacional Mayor

de San Marcos (MUSM). In this paper, this new species

of Petracola is described based on morphological data,

increasing the number of species to five.

Materials and Methods

Specimens and ecological data were collected in Rio

Abiseo National Park in 1987 by the first author (LOR).

Specimens were collected by hand, euthanized, fixed

in 10% formalin, and later transferred to 70% ethanol.

Terminology for diagnostics and descriptions follows

Kizirian (1996) and Goicoechea et al. (2013). Measure-

ments were taken with calipers accurate to 0.1 mm. Data

for other species were taken from the literature (Eche-

varria and Venegas 2015; Kizirian et al. 2008; Kohler

and Lehr 2004) and by examination of specimens de-

posited in the Museo de Historia Natural de la Universi-

dad Nacional de San Marcos (MUSM), Lima, Peru, and

the Museo de Biodiversidad del Peru (MUBI), Cusco,

Peru (Appendix I).

March 2020 | Volume 14 | Number 1 | e227

Rodriguez and Mamani

Results

Generic Assignment

The new species is assigned to Petracola based on the

presence of imbricate and scale-like papillae on the

tongue, head scales without striations or rugosities,

smooth and juxtaposed dorsal scales, and the absence of

prefrontal scales (Doan and Castoe 2005).

Taxonomy

Petracola pajatensis sp. nov.

urn:Isid:zoobank.org:act: E41 CD69F-3F7A-41A F-B756-BF836A536EA8

Holotype. MUSM 3829, adult male (Figs. 1—2) from Los

Chochos, Rio Abiseo National Park, Provincia Mariscal

Caceres, Departamento San Martin, Peru, approximately

3,230 m asl, 18 km airline from Pataz (7°38713’S,

77°28’ 80”"W), collected by Lily O. Rodriguez on 12 July

1987.

Paratypes. Three adult females (MUSM 3830 [Fig. 2],

15986-87), and one subadult male (MUSM 15985),

same data as holotype.

Diagnosis. (1) Frontonasal and frontal scales sub-equal:

(2) nasoloreal suture present, loreal scale not in contact

with supralabials; (3) supraoculars two; (4) superciliaries

two, discontinuous, first expanded onto dorsal surface of

head; (5) postoculars two; (6) palpebral disc divided in

two; (7) three supralabials anterior to the posteroventral

angle of subocular; (8) two pairs of genials in contact;

Amphib. Reptile Conserv.

Fig. 2. Dorsal, lateral, and ventral views of head of the holotype

(MUSM 3829).

March 2020 | Volume 14 | Number 1 | e227

New Petracola species from Peru

Fig. 3. Dorsal and ventral views of the paratype of Petracola pajatensis sp. nov. (MUSM 3830; female, SVL = 65.1, Tail = 64.3 mm).

(9) dorsal body scales quadrangular, smooth, juxtaposed;

(10) transverse dorsal rows 33-35; (11) transversal

ventral rows 20-22: (12) a continuous series of small

lateral scales separates dorsals from ventrals; (13)

posterior cloacal plate scales 2—5; (14) anterior preanal

plate scales paired; (15) femoral pores per hind limb in

males 6—8, in females three; (16) preanal pores absent:

(17) subdigital lamellae on finger IV eight; (18) limbs

not overlapping when adpressed against body on adults;

(19) pentadactyl, digits clawed; (20) coloration in liquid

preservative: in males, dorsum is light-brown with

numerous irregular dark-brown spots and venter is dark-

brown with some small cream spots on their flanks; in

females, dorsum is light-brown with some and irregular

dark-brown spots, venter is brown with cream spots that

form discontinuous transversal bands from the chest to

the anal plate (Fig. 3).

Petracola pajatensis sp. nov. can be distinguished

from P. angustisoma by having a robust body, two

discontinuous superciliaries, 6-8 femoral pores per

hind limb in males, maximum SVL in males 60.5

mm, dorsum is light-brown with irregular dark brown

spots not forming longitudinal stripes, venter is brown

in preservative (gracile body, three discontinuous

superciliaries, nine femoral pores per hind limb in males,

maximum SVL in males 43.6 mm, dorsum 1s brown or

olive with seven discontinuous dark brown longitudinal

stripes, venter is white with black semicircular black

spots on anterior margin of scales); from P. labioocularis

by having two supraoculars, absence of precloacal pores,

and two pairs of genials in contact (three supraoculars,

presence of precloacal pores, and usually three pairs of

genials in contact); from P. ventrimaculatus by having

6-8 femoral pores in males, in preservative the venter

Amphib. Reptile Conserv.

in males is dark brown with small lateral cream spots, in

females it is a combination of cream with brown forming

longitudinal bands, and maximum SVL in males 60 mm

(2-5 femoral pores in males, in males and females the

venter is cream with a bold black transversal band, and

maximum SVL in males 71.05 mm); from P. waka by

having two discontinuous superciliaries, two genitals in

contact, and venter in males is a dark brown with lateral

cream spots, in females the venter is a combination of

cream with brown forming a longitudinal band (four

continuous superciliaries, three genials in contact, and the

venter in males is white to pale yellow with brown spots).

Description of the holotype. Adult male, snout-vent

length (SVL) 60.5 mm, tail length 76.8 mm, head

scales smooth, rounded in lateral and ventral views,

without striations or rugosities; rostral scale wider

than tall, in contact with frontonasal, nasals, and first

supralabials; frontonasal longer that wide, longer

that the frontal scale, widest posteriorly, in contact

with rostral, nasal, first superciliary, and frontal;

prefrontal absent; frontal longer that wide, pentagonal,

in contact with first superciliary, first supraocular,

and frontoparietal; frontoparietal paired, polygonal

(hexagonal), in contact with frontal, supraoculars,

parietals, and interparietals; supraoculars two, the first

separates the first and second superciliaries, in contact

with superciliaries, frontal, frontoparietals, interparietal,

and postoculars; parietals longer than wide, polygonal

(irregular hexagon), in contact with frontoparietals and

supraoculars anteriorly, with interparietal, and temporals

laterally, and with postparietals posteriorly; interparietal

polygonal (hexagonal), in contact with frontoparietals

anteriorly, with parietals laterally, and with postparietal

March 2020 | Volume 14 | Number 1 | e227

Rodriguez and Mamani

posteriorly; postparietals paired, smaller than parietals,

and polygonal. Nasal scale divided, longer than high,

in contact with first supralabials; loreal scale present

on the left side, nasoloreal suture incomplete on the

right side, not in contact with the supralabials; two

superciliaries, discontinuous, and first expanded onto the

dorsal surface of the head; two preoculars; frenocular

fused with the first subocular only on the right side;

palpebral disc transparent and divided in two; three

suboculars, on the right side the first subocular is fused

with the frenocular; two postoculars; temporals smooth,

polygonal; three supralabials anterior to posteroventral

angle of suboculars. Mental wider than long, in contact

with first infralabials and postmental posteriorly:

postmental single, polygonal (irregular heptagonal), in

contact with the first and second infralabials, and the

first pair of genials; 3/2 genials, on right and left sides

respectively, all in contact medially, the first genial on

right side 1s divided; three transversal rows of pregular

scales; six gular scale rows, polygonal, and smooth.

Dorsal scales rectangular, longer that wide, juxtaposed,

smooth, 33 transverse rows; 21 longitudinal dorsal scale

rows at midbody; a continuous series of small lateral

scales; reduced scales at limb insertion region present;

21 transverse ventral scale rows; 10 longitudinal ventral

scale rows at midbody, the lateral scales are slightly

smaller; anterior preanal plate scales paired; three

posterior preanal plate scales, and a small and polygonal

scale lies between the anterior and posterior preanal

plate scales; scales on the tail rectangular, juxtaposed,

and smooth; ventral scales quadrangular, juxtaposed,

and smooth. Limbs pentadactyl; digits clawed; dorsal

brachial scales polygonal, subimbricate, and smooth:

ventral brachial scales rounded, subimbricate, and

smooth; dorsal antebrachial scales polygonal, smooth;

ventral antebrachial scales polygonal, smooth, smaller

than dorsal; dorsal manus scales polygonal, smooth,

subimbricate; palmar scales small, rounded, juxtaposed,

and domelike; dorsal scales on fingers smooth, quad-

Amphib. Reptile Conserv.

rangular, imbricate, three on finger I, five on II, six on III,

five on IV, and four on V; scales on anterodorsal surface

of thigh polygonal, smooth, subimbricate; scales on

posterior surface of thigh small, rounded, and juxtaposed;

scales on ventral surface of thigh small, polygonal and

juxtaposed; six femoral pores on left thigh and seven

on right; scales on anterior surface of crus polygonal,

smooth, juxtaposed, decreasing in size distally; scales

on posterodorsal surface of crus smooth, polygonal,

juxtaposed; scales on ventral surface of crus polygonal,

enlarged, smooth, and subimbricate; scales on dorsal

surface of toes polygonal, smooth, and subimbricate;

scales on ventral surface of toes rounded, small, and

domelike; dorsal scales of toes smooth, imbricate, two

on toe I, five on toe II, seven on toe III, ten on toe IV, and

six on toe V.

Coloration

In preservative. Petracola pajatensis sp. nov. exhibits

a variable coloration in adults of both sexes. The males

(Fig. 1) have a light-brown dorsum with numerous and

irregular dark-brown spots, with a cream continuous

dorsolateral line on both sides of the body that starts

from the back of the eye to the tail. This cream line 1s

bordered on both sides by a continuous dark-brown line;

the venter is dark-brown with some small cream spots

on their flanks. The females (Fig. 3) have a light-brown

dorsum with some irregular dark-brown spots, with a

cream discontinuous dorsolateral line on both sides of the

body that starts from the back of the eye to the tail; the

venter is brown with cream spots that form continuous

longitudinal bands from the chest to the anal plate; the

neck and chin are a combination of cream and brown.

In life. Based on unvouchered specimens, the dorsal

coloration is brown with irregular dark spots distributed

irregularly, the dorsolateral lines are obscure, and the

flanks have more dark spots than the dorsum (Fig. 4).

March 2020 | Volume 14 | Number 1 | e227

New Petracola species from Peru

Fig. 5. Drawings of the right side showing the condition of loreal

scales in Petracola pajatensis sp. nov. A) MUSM 3829, B)

MUSM 3830.

Variation

Considerable variation is evident in the position

and form of the loreal scale. The loreal scale can be

incomplete, tiny, or enlarged; and can be separating, or

not, the first superciliary and nasal scale; in the holotype

(MUSM 3829) the right scale is incomplete and the left

does not separate the nasal and first superciliary, while

in three paratypes (MUSM 3830, 15985-6) the loreal

scale separated the nasal and first superciliary; and in

one paratype (MUSM 15987) the first superciliary and

nasal are in contact on both sides (Figs. 2, 5). Variation

in meristic characters is shown in Table 1. No evidence

of sexual dimorphism exists in the scutellation, except

for the larger number of and better developed femoral

pores in males.

Etymology

The specific epithet is an adjective that recognizes Gran

Pajatén archaeological remains, which, like Petracola

pajatensis sp. nov., occurs in Rio Abiseo National Park.

Habitat, Ecological Notes, and Distribution

The holotype was taken from the ground in grassland, ina

rocky area dominated by bunchgrasses of Calamagrostis

Amphib. Reptile Conserv.

Fig. 6. Type locality of Petracola pajatensis sp. nov. Photo by

Ken R. Young.

81°0'0"O

78°0'0"O 75°0'0"O 72°0'0"O 69°0'0"O

Legend

Altitude Pacific

Value

aa 2 Ocean

HB 200-00

HE 200-1000

GE 1000-1500

[HB 1500-2000

[J] 2000-2800

[EJ 2500-3000

[0 3000-3500

HB 3500-4000

GE 2000-4500

; | > 4500

81°0'0"O.

0 62.5125 250 375 500

ee Kn

78°0'0"O 75°0'0"O 72°0'0"O 69°0'0"O

Fig. 7. Locations of the type localities of the species of Petracola

(P. angustisoma = circle, P. ventrimaculatus = square, P. waka

= diamond, P. /abioocularis = triangle, P. pajatensis sp. nov.

= star). Sources: Echevarria and Venegas (2015), Echevarria

(2014), Kizirian et al. (2008), and Kohler and Lehr (2004).

sp., Festuca sp., Cortaderla sp., and Stipa sp. (Fig.

6). The habitat of Petracola pajatensis sp. nov. was

previously disturbed through overgrazing and 1s

currently undergoing secondary succession, including

scattered shrubs (Baccharis). Multiple nest deposition

sites contained elliptical egg shells (9.5—11.0 mm). The

new species is known only from Los Chochos, a small

valley on the western slopes of the Cordillera Oriental,

south of the Huancabamba Depression, approximately

100 km south of the localities of P. ventrimaculatus

(Fig. 7).

March 2020 | Volume 14 | Number 1 | e227

Rodriguez and Mamani

Table 1. Measurements and scale counts of specimens of Petracola pajatensis sp. nov. and P. ventrimaculatus examined in this

paper. *only an adult male, ** the first right genial is divided, ***the anterior subocular scale is fused with freno ocular.

aca ieieiat | ae

Characters (count or mm)

Supralabial scales to

posteroventral angle of 3.0 3.0 3.0 3—4

subocular

[Gularsialerows [| 60 -+[ 56 | 60 | 60

Longitudinal dorsal scale 17-21 16-20 95295 20-23

rows

Transversal ventral scale

rows

Lamellae under 4" finger

Lamellae under 4" toe 14-15 13-14 12-13 14-15

Acknowledgments.—We are grateful to the Direccion

General de Fauna Silvestre for kindly providing special

collecting permits for the specimens from Abiseo

National Park (N° 18-87-AG-DGFF-SDFF). Cesar

Aguilar (MUSM) and Juan Carlos Chaparro (MUBI)

provided access to herpetological collections. D.

Aguilar and M. Silva assisted LOR in the field. The

Packard Foundation and the Pew Charitable Fund

funded, through APECO, the faunal inventory at the

Abiseo National Park. The BIOLAT program of the

Smithsonian Institution provided support for collection

studies at the National Museum in Washington, DC,

USA, where Tom Fritts kindly provided guidance to

LOR to initiate this description and C. Myers from the

American Museum provided working space for LOR.

Ken R. Young kindly provided us with photos from the

type locality and a live specimen (Figs. 3-4). Finally,

We thank the anonymous reviewers for their valuable

comments and suggestions on the manuscript.

Amphib. Reptile Conserv.

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1900 utilizando evidencias morfoldgicas y ecoldgicas.

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of Petracola (Squamata: Gymnophthalmidae) from

the Utcubamba Basin in the Andes of northern Peru.

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Section]: 26—33 (e107).

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Fisher S, De la Riva I. 2013. A taxonomic revision

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New Petracola species from Peru

of Proctoporus bolivianus Werner (Squamata:

Gymnophthalmidae) with the description of three

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nine new species. Herpetological Monographs 10:

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Donnelly M. 2008. A new Petracola and a re-

description of P ventrimaculatus (Squamata:

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Peru, with the description of three new species and

a key to the Peruvian species. Herpetologica 60(4):

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Cordillera de los Andes.

Appendix 1. Specimens examined.

Petracola waka

Moravec J, Smid J, Stundl J, Lehr E. 2018. Systematics of

Neotropical microteiid lizards (Gymnophthalmidae,

Cercosaurinae), with the description of a new genus

and species from the Andean montane forests.

ZooKeys 774: 105-139.

Torres-Carvajal O, Lobos SE, Venegas PJ, Chavez G,

Aguirre-Pefiafiel V, Zurita D, Echevarria LY. 2016.

Phylogeny and biogeography of the most diverse

clade of South American gymnophthalmid lizards

(Squamata, Gymnophthalmidae, Cercosaurinae).

Molecular Phylogenetics and Evolution 99: 63-75.

Uzzel TM. 1970. Teiid lizards of the genus Proctoporus

from Bolivia and Peru. Postilla 142: 1-39.

Vasquez-Restrepo JD, Ibafiez R, Sanchez-Pacheco

SJ, Daza JM. 2019. Phylogeny, taxonomy, and

distribution of the Neotropical lizard genus

Echinosaura (Squamata: Gymnophthalmidae), with

the recognition of two new genera in Cercosaurinae.

Zoological Journal of the Linnean Society 20: 1—28.

Lily O. Rodriguez is a herpetologist, holding a doctoral degree in Ecology from the Université de

Paris. With extensive fieldwork in the Peruvian Amazon, Lily has a special interest in the diversity and

distribution of herpetofauna across Peru. She is currently working for the Center for Conservation,

Research, and Management of Natural Areas, CIMA-Cordillera Azul and is a Research Associate

to the Museo de Historia Natural of the Universidad Nacional Mayor de San Marcos in Lima, Peru.

Luis Mamani is a biologist who graduated from the Universidad Nacional de San Antonio Abad

del Cusco (Peru) and he obtained his M.Sc. degree from the Universidad de Concepcion (UdeC) in

Chile. Currently, Luis is a researcher at Museo de Biodiversidad del Peru (MUBIJ) and Museo de

Historia Natural de la Universidad Nacional de San Antonio Abad del Cusco (MHNC). His current

research includes systematics, taxonomy, and biogeography of the gymnophthalmid lizards from the

PERU: Cajamarca: Bafios del Inca: MUBI 4721, 4722, 4723; Cajabamba: MUBI 2609-2619, KU 135063-79;

Celendin: El Sauce: MUSM 26246-26248, 26346-26351.

Petracola ventrimaculatus

PERU: Cajamarca: Celendin: MUBI 11114, 11117, 1118; Hualgayoc: MUBI 11402, 11403.

Petracola labioocularis

PERU: Huanuco: Monte Potrero: MUBI 14252, 14253, 14254, 14232, 14251; Acomayo: MUSM 13903, 13904.

Petracola angustisoma

PERU: Amazonas: Bongara: San Carlos: Achupampa: MUBI 11513.

Amphib. Reptile Conserv.

146

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Official journal website:

amphibian-reptile-conservation.org

Amphibian & Reptile Conservation

14(1) [General Section]: 147-155 (e228).

The herpetofauna of Honaz Mountain National Park

(Denizli Province, Turkey) and threatening factors

*Miige Gidig and 2Eyup Baskale

'Kiitahya Dumlupinar University, Faculty of Arts and Science, Department of Biochemistry, Kiitahya, TURKEY ?Pamukkale University, Faculty of

Arts and Science, Department of Biology, Denizli, TURKEY

Abstract.—The aim of this study was to catalog the species of amphibians and reptiles in Honaz District, in

the province of Denizli, Turkey, based on field surveys carried out during March—October of 2015-2018. The

species found include five amphibians (Bufo bufo, Bufotes variabilis, Pelophylax bedriagae, Hyla orientalis,

Rana macrocnemis) and 22 reptiles: two turtles (Wauremys rivulata, Testudo graeca), nine lizards (Stellagama

stellio, Mediodactylus kotschyi, Anatololacerta danfordi, Lacerta trilineata, Ophisops elegans, Ablepharus

kitaibellii, Pseudopodus apodus, Blanus strauchi, Heremites auratus), and 11 snakes (Xerotyphlops

vermicularis, Dolichophis jugularis, D. caspius, Hemorrhois numnifer, Eirenis modestus, Telescopus fallax,

Elaphe sauromates, Natrix natrix, N. tessellata, Eryx jaculus, Montivipera xanthina). In addition, the various

factors threatening these species in this area were determined and proposals for the conservation of these

species are presented.

Keywords. Amphibians, Anura, distribution, habitat destruction, pollution, reptiles, Squamata, Testudines

Citation: Gidis M, Baskale E. 2020. The herpetofauna of Honaz Mountain National Park (Denizli Province, Turkey) and threatening factors. Amphibian

& Reptile Conservation 14(1) [General Section]: 147-155 (e228).

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: 19 April 2019; Accepted: 5 October 2019; Published: 3 April 2020

Introduction

Amphibians and reptiles are an important component

of most ecosystems. In particular, their importance 1s

incontrovertible for the maintenance of food networks.

Their prey includes a variety of insects and their larvae,

fish, shellfish, other reptiles, other amphibians, birds,

and small mammals. They are also a source of food

for fish, birds, reptiles, and mammals. From this point

of view, amphibians and reptiles play an important role

in ensuring the balance within ecosystems (Bohm et

al. 2013; Heatwole and Wilkinson 2015). In addition,

because they are poikilotherms and as they are in close

contact with the air, water, and soil according to the

characteristics of the habitats in which they are located,

they have good characteristics as indicators for food

networks (Duellman and Trueb 1994) and environmental

changes (Demirsoy 1996; Baran and Atattir 1998; Budak

and Go¢men 2008; Baran et al. 2012). In recent years,

studies focusing on the assessment of species diversity

among ecological communities have been instrumental

in developing protection strategies, as well as helping

to clarify the structure and dynamics of biological

communities (Baldwin et al. 2006; Shaney and Marshall

2013; Berriozabal-Islas et al. 2017; Curi¢ et al. 2018).

Correspondence. *mugegidis@gmail.com

Amphib. Reptile Conserv.

Amphibians and reptiles are two of the most

endangered groups compared to other vertebrates

(Hoffmann et al. 2010). The herpetofauna of Turkey

has been studied intensively in the last 50 years,

providing information about the distribution of the

many amphibian and reptile species in Turkey (Sillero

et al. 2014; Yildiz and I&ci 2015; Bulbul et al. 2016;

Go¢men and Karis 2017; Sarikaya et al. 2017; Gul

et al. 2018; Yildiz et al. 2019). However, due to

technological and methodological developments the

taxonomy of reptiles and amphibians has remained

dynamic and unstable. For example, in 2008, only

8,734 reptile species were reported for the world, but as

of July 2018 this figure had increased to 10,793 (Reptile

Database: http://www.reptile-database.org). Amphibian

species are represented by about 7,950 species globally

(AmphibiaWeb: http://www.amphibiaweb.org), and

from 2004 through November 2018, a total of 2,318

new amphibian species were described, including 139

from January 2018 until 14 November 2018. A similar

situation is found in Turkey, where the number of

known amphibian and reptile species was 141 in 2013,

and with the latest developments, the number of species

has grown to 170 (Sindaco et al. 2000; Yildiz and [&ci

2015; Bulbul et al. 2016; Yildiz et al. 2019).

April 2020 | Volume 14 | Number 1 | e228

Herpetofauna of Honaz Mountain National Park, Turkey

The history of studies on the Turkish herpetofauna

traces back to the 19th century (Boettger 1889), and in

recent years many new species have been identified and

new records have been published (Sillero et al. 2014;

Yildiz and [%ci 2015; Bulbul et al. 2016; Wielstra and

Arntzen 2016; Gocmen and Karis 2017; Sarikaya et

al. 2017; Gul et al. 2018; Yildiz et al. 2019). Faunistic

inventory studies in Denizli province were conducted

in 2013 by the Ministry of Forestry and Water Affairs,

General Directorate of GDMP, and the Pamukkale

University Biology Department. However, that study

did not provide much information about the species and

the factors that threaten them in the province of Denizli.

The current survey (conducted from 2015-2018) focused

on the herpetological species of the Honaz Mountain

National Park-Denizli, which is one of Turkey's most

important protected areas, and elucidating the threatening

factors for amphibian and reptile species living in this

national park. The results provide a new opportunity for

revising the conservation status of the Honaz Mountain

National Park-Denizli herpetofauna and a new reference

for distributional data.

Materials and Methods

Honaz Mountain National Park-Denizli, which is located

within the borders of the Honaz district of Denizli

province, was declared a National Park on 21 April

1995 and approved by the General Directorate of Nature

Conservation National Parks on 9 March 2009. The area

of the national park is 9,616 ha and the highest point has

an altitude of 2,571 m. The climate is dominated by the

Mediterranean climate and the park is mostly covered

with Red Pine (Pinus brutia) forest. A map of the Honaz

Mountain National Park and the survey sites is given in

Fig. 1.

Field surveys were conducted between March and

October in each year from 2015 to 2018. Amphibians and

reptiles in this area are active during spring and summer,

which is the best time to study them. Early autumn can

also be considered suitable for observing reptiles before

hibernation. The Visual Encounter Survey (VES, Crump

and Scott 1994) method was used in the field studies to

assess the amphibian and reptile species. In this study,

two teams conducted the field studies along linear paths

according to the habitat characteristics, such as forest

land, stony areas, and bush. The specimens captured

during the surveys were diagnosed, and assessed at the

species or family levels. According to the size of the

habitat, 0.5—1.5 h of fieldwork was performed at each

station. Specimens which could not be identified in the

field were transported to the laboratory, and species were

diagnosed by using an identification key or the current

literature for the relevant species (Baran and Atatiir

1998; Ozeti and Yilmaz 1994; Baran et al. 2012; Budak

and Gocmen 2008).

Amphib. Reptile Conserv.

8@ Honaz Mountain National Park 12@

14@@"?

a 15@

0 1,25 2,5

Kilometers

Fig. 1. Study area and locations of field surveys in Honaz

Mountain National Park, Turkey. The coordinates and elevations

of the numbered sites are given in Appendix 1.

After the species were identified, their population

status was further determined using the following

resources: 1) Convention on the Conservation of

European Wildlife and Natural Habitats = BERN

(Bern 1982), 2) Convention on International Trade in

Endangered Species of Wild Fauna and Flora (CITES

1975), and 3) The International Union for Conservation

of Nature’s Red List of Threatened Species = IUCN Red

List (http://www.redlist.org). Additional information was

gathered on the factors that threaten the populations in

the survey areas.

Results

In this study, a total of 27 amphibian and reptile

species belonging to 13 families were observed in

Honaz Mountain National Park (Fig. 2). The species of

amphibians and reptiles found in the research area, and

their associated habitat types, are given in Tables 1 and 2.

According to the Bern Convention, 12 of the amphibian

and reptile species are listed in Appendix 2, and 15 species

are on the Annex 3 list. According to the IUCN Red List

criteria, one species is listed as VU (Vulnerable), 21 as

LC (Least Concern), and one as DD (Data Deficient)

[Fig. 3A]. Four species have not been evaluated yet (NE),

April 2020 | Volume 14 | Number 1 | e228

Gidis and Baskale

ete 4

Fig. 2. Some of the amphibian and reptilian species in the study

nn * Pe Ra:

wt We

~*~

area: (a) Bufo bufo, (b) Hyla orientalis, (c) Rana macrocnenmis,

(d) Testudo graeca, (e) Mediodactylus kotschyi, (f) Ablepharus kitaibellii, (g) Stellagama stellio, (h) Anatololacerta danfordi, (i)

Blanus strauchi, (j) Eryx jaculus, (k) Natrix natrix, and (1) Montivipera xanthina.

so they are not listed. In addition, according to CITES,

two species are included in the Appendix 2 list and 25

are excluded from the list. The chorotype distributions

of the species indicated that the amphibian and reptile

species of Honaz Mountain National Park mostly

belonged to the Turanian-Mediterranean and Eastern

Mediterranean chorotypes (Vigna Taglianti et al. 1999;

Sindaco et al. 2000) [Fig. 3B]. This study will inform the

conservation of amphibian and reptile species in this area

and contributes to the broader knowledge of Turkey's

herpetofauna.

Discussion

Turkey has a rich flora and fauna due to its geological,

topographical, andclimaticcharacteristics. Herpetological

Amphib. Reptile Conserv.

studies conducted for more than a century show that there

is not enough information available on the herpetofauna

of Turkey. On the one hand, in the last 20 years, new

species and subspecies have been uncovered in Turkey

and new localities have been recorded for the existing

species (Ayaz et al. 2011; G6¢men et al. 2013; Avci et al.

2015; Uztim et al. 2015; Bulbiil et al. 2016). On the other

hand, herpetology research has been limited to certain

areas in Turkey despite the high number of divergent

areas with differences in geographic isolation elements,

and different ecosystems and ecological conditions.

As a result, in the last 20 years, studies determining

the amphibian and reptile species and their ecological

preferences in the previously understudied regions have

accelerated (Ozdemir and Baran 2002; Afsar and Tok

2012; Ozcan and Uztim 2013; Cihan and Tok 2014; Ege

April 2020 | Volume 14 | Number 1 | e228

Herpetofauna of Honaz Mountain National Park, Turkey

Table 1. Amphibian species found in Honaz Mountain National Park and their characteristics.

Habitat IUCN | Population status Chorotype CITES

Forestlands,

waysides,

waterfronts,

rocky terrains,

meadowlands,

croplands, gardens

Bufonidae | Bufo bufo

(Linnaeus, 1758)

Bufonidae | Bufotes variabilis

(Pallas, 1769)

Forestlands,

waterfronts,

meadowlands,

croplands, gardens

Forestlands,

waysides,

waterfronts,

croplands

Ranidae Pelophylax

bedriagae

(Camerano,

1882)

Ranidae Rana

macrocnemis

(Boulenger,

1885)

Hylidae Hyla orientalis

(Bedriaga, 1890)

et al. 2015; Eksilmez et al. 2017; Kumlutas et al. 2017;

Sarikaya et al. 2017).

Most studies on the herpetofauna of Denizli

province were carried out in the individual districts.

For example, 17 reptile and amphibian species were

found in the Babada§g district (Urhan et al. 1999); 24 in

Acipayam-Denizli (Urhan et al. 2003); 26 in Hambat

(Urhan et al. 2004); and 22 around the Cal, Bekilli, and

Baklan Districts (Urhan et al. 2006). Unal et al. (2012)

identified 25 amphibian and reptile species belonging to

15 families in Kale district. In this study, five anurans,

two turtles, nine lizards, and 11 snakes (for a total of

five amphibian and 22 reptile species) were observed

in Honaz Mountain National Park. There are no current

taxonomic controversies among these species. A

previous study conducted about 20 years ago reported

21 amphibian and reptile species from Honaz Mountain

National Park (Katilmis et al. 2002). In this study, Hyla

orientalis, Rana macrocnemis, Mauremys_ rivulata,

Heremites auratus, Pseudopodus apodus, and Telescopus

fallax were recorded for the first time in Honaz Mountain

National Park. In addition, seven amphibian and 26

reptile species have been identified in Denizli province

during 2002—2009 (Urhan et al. 2009). In this respect,

the Honaz Mountain National Park includes 71.4% of the

amphibian diversity and 80.8% of the reptile diversity in

Denizli province.

Of the ~18,000 amphibian and reptile species

distributed around the world, about 35% are on the IUCN

Red List. According to IUCN criteria, among the total of

27 amphibian and reptile species found in the study area:

one (4.76%) 1s Vulnerable (VU), 21 (66.7%) are Least

Concern (LC), one (4.76%) is Data Deficient (DD), and 4

(28.57%) are not evaluated yet IUCN 2018). According

to the IUCN Red List, the populations of 10 of these

Waterfronts,

croplands

Forestlands,

Amphib. Reptile Conserv.

Undetermined/

unknown

L Declining

maquis shrublands,

shrubbery

132-98" 10;

2 > >

17, 18, 19

a _

fa al a

species are stable (37.04%), while the populations of

four species are decreasing (14.81%), and there is no

information on the population status of the remaining 13

species (48.15%). Therefore, it is necessary to determine

the roles of various threats to the ecosystem and the

current population status of the species of amphibians

and reptiles that were found.

Turano-Europeo-

Mediterranean

Turano-Europeo-

Mediterranean

SW-Asiatic

European I

Mediterranean,

8.3%

European,

8.3%

Turano-Europeo-

Mediterranean, 4.2%

NAA SE-Anatolia- SW-Asiatic, 16.7%

erie Endemic, 4.2%

Fig. 3. Distribution of species according to IUCN categories

(A) and their chorotypes (B).

Turano-

Mediterranean,

25.0%

Central Asiatic-

Europeo-

Mediterranean, 4.2%

E. European,

25.0%

April 2020 | Volume 14 | Number 1 | e228

Gidis and Baskale

Table 2. Reptile species found in Honaz Mountain National Park and their characteristics.

Testudinidae Testudo graeca | Forestland, waterfronts, VU Stable Turano- 1, 3, 6, 8,

(Linnaeus, rocky terrain, meadow- Mediterranean Is, £6, TF,

1758) land, maquis shrubland,

cropland, gardens, moor-

land

Anguidae Pseudopus Forestland, maquis shru- Undetermined/ Turano-

apodus (Pallas, | bland, cropland, gardens

1775)

Unknown Mediterranean

Geoemydidae | Mauremys Waterfronts, creeks

rivulata (Va-

lenciennes in

Bory de Saint-

Vincent, 1833)

Agamidae Stellagama Wayside, rocky terrain

stellio (Lin-

naeus, 1758)

Gekkonidae Mediodactylus_ | Forestlands, waysides,

kotschyi (Stein- | open slopes, valley slopes,

dachner, 1870) | rocky terrains, maquis

shrublands, croplands,

gardens, residental areas

Lacertidae Anatololacerta | Forestlands, waterfronts,

danfordi (Giin- | open slopes, valley slopes,

ther, 1876) rocky terrains, meadow-

lands, maquis shrublands,

croplands

Undetermined/ Turano-

Unknown Mediterranean

Stable E.

Mediterranean

Undetermined/ E.

Unknown Mediterranean

Stable SE-Anatolia-

Endemic

Undetermined/ E.

Unknown Mediterranean

Stable E.

Mediterranean

—

Undetermined/ | Mediterranean

Unknown

Lacertidae Ophisops Forestland, rocky terrains,

elegans (Méné- | croplands, bushlands,

tries, 1832) moorlands, semi-desert

areas

Scincidae Ablepharus Forestlands, rocky ter-

kitaibelii (Bi- rains, meadowlands

bron and Bory

St-Vincent,

1833)

Scincidae Heremites Rocky terrains

auratus (Lin-

naeus, 1758)

Amphisbae- Blanus strau- Forestlands, rocky ter-

nidae chi (Bedriaga, | rains, croplands

1884)

Typhlopidae | Xerotyphlops Forestlands, waterfronts,

vermicularis rocky terrains, meadow-

(Merrem, 1820) | lands

Colubridae Dolichophis Waterfronts, open slopes,

Jugularis (Lin- | valley slopes, rocky ter-

naeus, 1758) rains, cropland, gardens

Colubridae Dolichophis Open slopes, rocky ter-

caspius (Gme- | rains, meadowlands,

lin, 1789) croplands, gardens

Colubridae Hemorrhois Open slopes, rocky ter-

nummifer (Re- | rains

uss, 1834)

Colubridae Eirenis modes- | Rocky terrains Stable SW-Asiatic

tus (Martin,

1838)

Amphib. Reptile Conserv. 151 April 2020 | Volume 14 | Number 1 | e228

Undetermined/ Turano-

Unknown Mediterranean

—

Undetermined/ Turano-

Unknown Mediterranean

—

Undetermined/ Turano-

Unknown Mediterranean

Lacertidae Lacerta trilin- | Forestlands, rocky ter- Stable iE.

eata (Bedriaga, | rains, croplands, bush- Mediterranean

1886) lands, moorlands, semi-

desert areas

Herpetofauna of Honaz Mountain National Park, Turkey

Table 2 (continued). Reptile species found in Honaz Mountain National Park and their characteristics.

Colubridae Telescopus Forestlands, bushlands, LC

fallax (Fleis- rocky terrains

chmann, 1831)

Colubridae Elaphe sauro-

mates (Pallas,

1811)

Forestlands, waysides,

open slopes, rocky ter-

rains, meadowlands, crop-

lands, gardens, bushlands,

moorlands

Forestlands, waterfronts,

meadowlands

Colubridae Natrix natrix

(Linnaeus,

1758)

Colubridae Natrix tessel- Waterfronts

lata (Laurenti,

1768)

Boidae Eryx jaculus Forestlands, open slopes,

(Linnaeus, valley slopes, rocky ter-

1758) rains

Viperidae Montivipera Open slopes, valley

xanthina (Gray, | slopes, rocky terrains

1849)

As indicated by field studies, all of the threats to

amphibian and reptile diversity are found to originate from

anthropogenic degradation of the environment (Curic et

al. 2018; Hosseinzadeh et al. 2018). Destruction of habitat

is the major threat for many species. Considering that the

terrestrial frog and reptile species are mainly distributed

in mountain slopes, as well as rocky and stony areas,

mining activities pose a significant threat to biodiversity.

Destruction and restriction of the habitats of amphibian

and reptile species may cause them to become trapped

in certain areas over the long term. The competition of

species having a common niche will cause stress in terms

of food resources and reproduction, and species that are

trapped in certain areas will be more sensitive to changes

in the ecosystem. Two other problems are the agricultural

activities and domestic pollution which affect the species

that are dependent on aquatic habitats. The species

living in irrigation channels in agricultural lands are

directly exposed to pesticides. During our field studies,

packages of agricultural pesticides were found in many

habitats, such as irrigation channels and streams. Another

important threat for the reptiles is that local people do

not recognize the non-venomous reptile species and kill

reptiles indiscriminately due to mis-information. Most

snake species encountered in the field or in different

environments are killed despite being harmless. Even the

European Glass Lizard (Pseudopodus apodus) is known

by the local people as the “iron snake” and it is often

killed due to the belief that it is venomous. As reptiles do

not have mimic muscles and they have scaly structures,

people often consider them to be “cold-faced animals.”

Another problem observed in the survey area is that

amphibians and reptiles are often crushed by road traffic.

Several recommendations for the conservation of

species are given here.

Amphib. Reptile Conserv.

Declining

Undetermined/ Turano-

Unknown

Mediterranean

Undetermined/ Turano-

Unknown

Europeo-

Mediterranean

Undetermined/ | Central Asiatic-

Unknown

Europeo-

Mediterranean

Central Asiatic-

European

Undetermined/ | Mediterranean

Unknown

Stable E.

Mediterranean

¢ Information and training activities should be initi-

ated in the habitat areas which are intensively used

by people, such as recreational areas. In addition,

Natural Life Protection Schools, supported by various

institutions and associations, should be expanded and

brought to a level that will appeal to more people.

¢ Modification or destruction of habitats should be

prevented. If a study on habitat is conducted, at a

minimum it should be carried out under the supervi-

sion of a commission of biologists and should be done

outside of the breeding season.

* Most specimen mortalities occur during the migra-

tion to wetland ecosystems during the reproductive

season. During migration, many adult individuals are

destroyed on the roads due to heavy traffic. For this

reason, barriers should be built around the main arter-

ies in the wetland ecosystems and their channels; and

similar structures should be built to ensure that indi-

viduals migrating to the wetland can pass from safe

areas to the wetlands.

¢ Predator species not found in the natural environ-

ment should not be released to the wetland ecosys-

tems. Numerous studies have shown that species

added to the environment for biological control

are harmful to the ecosystem over time, and often

a struggle ensues to remove these species from the

environment. For this reason, steps should be taken

to ensure that these predators are removed as soon

as possible.

¢ Due to the agricultural activities around the wet-

lands, many chemical pollutants are introduced into

the water bodies. Therefore, when controlling harm-

ful plants and insects, only herbicides and insecticides

approved by the Republic of Turkey Ministry of Ag-

riculture and Forestry should be used, and disposal of

April 2020 | Volume 14 | Number 1 | e228

Gidis and Baskale

their waste materials should be in an isolated environ-

ment away from the water bodies.

¢ The environment is continuously being loaded with

chemical substances released by anthropogenic activi-

ties that may directly or indirectly affect herpetofau-

nal populations. The geno- and ecotoxicological ef-

fects of these substances on living things should be

further investigated.

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Denizli.

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Carpaneto GM, De Biase A, Venchi A, Zapporoli M.

1999. A proposal for a chorotype classification of the

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in Triturus ivanbureschi (Amphibia: Caudata:

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herpetofauna of the Province of Hatay (East

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Conservation 12(2): 197-205.

Miige Gidis is an Assistant Professor in the Department of Biochemistry at the University of

Dumlupinar (Kttahya, Turkey). Miige is interested in the molecular ecology and conservation

biology of reptiles and amphibians. Her recent research projects focus on the comparative

phylogeography of amphibians and reptiles.

Eyup Baskale is a Professor in the Department of Biology at the University of Pamukkale (Denizli,

Turkey). His current research seeks to understand animal ecology and distribution, with special

focus on the conservation of amphibians and reptiles living in Turkey. Eyup currently coordinates

several research projects focusing on threatened species in Turkey.

April 2020 | Volume 14 | Number 1 | e228

Appendix 1. Coordinates and altitudes of the study locations at Honaz Mountain National Park, Turkey.

Amphib. Reptile Conserv.

Locality no.

0 AN HNN FW WH

NO NON NO HHH RFR RR Re Rl

YN F& OO WAN WD nA FW NF CO

Gidis and Baskale

Longitude

29°15.646'E

29°19.080'E

29°15.999'E

29°14.361'E

29°17.545'E

29°14.031'E

29°14.692'E

29°18.739'E

29°18 .828'E

29°17 .338'E

29°16.884'E

29°17 3560'E

29°15.847'E

29°15.865'E

29°13.089'E

29°18.984'E

29°17.642'E

29°13.609'E

29°16.014'E

29°20.141'E

29°16.090'E

29°14.095'E

155

Latitude

37°47.216'N

37°44 .227'N

37°43.358'N

37°41.585'N

37°42.625'N

37°44 044'N

37°39.455'N

37°42.911'N

37°43. 486'N

37°42. 487'N

37°42.254'N

37°44 .199'N

37°41.047'N

37°39.939'N

37°44 .586'N

37°41.988'N

37°40.152'N

37°42.896'N

37°47.440'N

37°43.829'N

37°44 489'N

37°38.874'N

Altitude (m)

376

1,101

1,345

1,156

1,427

1,138

1,200

1,645

1,622

1,402

1,405

785

1,909

1,592

902

1,742

2,025

1,070

362

1,109

808

1,272

April 2020 | Volume 14 | Number 1 | e228

Amphibian & Reptile Conservation

14(1) [General Section]: 156-162 (e229).

Official journal website:

amphibian-reptile-conservation.org

Patterns of growth and natural mortality in Lysapsus

bolivianus (Anura, Hylidae, Pseudae) in an environmental

protection area in the estuary of the Amazon River

1* Julio C. Sa-Oliveira, ?Carlos E. Costa-Campos, *Andréa S. Araujo, and ‘Stephen F. Ferrari

'Research Nucleus in Fisheries and Aquaculture-NEPA, Laboratory of Limnology and Ichthyology, Federal University of Amapad (UNIFAP), Campus

Universitario Marco Zero do Equador, Rod. Juscelino Kubitscheck, Km 02, CEP 68903-419 Macapa, Amapd, BRAZIL *Herpetology Laboratory,

Federal University of Amapa (UNIFAP), Campus Universitario Marco Zero do Equador, Rod. Juscelino Kubitscheck, Km 02, CEP 68903-419

Macapa, Amapa, BRAZIL *Zoology Laboratory, Federal University of Amapd (UNIFAP), Campus Universitario Marco Zero do Equador, Rod.

Juscelino Kubitscheck, Km 02, CEP 68903-419 Macapd, Amapd, BRAZIL *Department of Ecology, Federal University of Sergipe — UFS, Sdo

Crist6vdo, BRAZIL

Abstract.—Recent reviews indicate that about one-third of amphibian species are threatened with extinction.

Many of these species inhabit tropical areas in developing countries where deforestation and the degradation of

natural bodies of water are major threats. Lysapsus bolivianus is a poorly known amphibian found throughout

much of the central Amazon basin between Bolivia and the Amazon estuary, where it is subject to extensive

anthropogenic pressures. The present study was based on samples of this species in an environmental

protection area. The data obtained is important for understanding the population structure with respect to size,

growth parameters (K, Ay 95° L_, 2’, SVL_...): and natural mortality of the species. The results showed a sexual

dimorphism in size, with females being larger. Both sexes presented fast growth rates (K,,,,,= 0.71 year’; K.._ =

0.70 year"), reaching asymptotic sizes (SVL~,,_,.= 21.20 mm; SVL~,,,__,,= 25.60 mm) in less than twelve months,

and longevity of <5 years. The species completes its metamorphosis in 20 days, reaching adult age at one

month. The estimated natural mortality was 0.64 year’ for males and 0.65 year" for females. The precocity of

this species, as well as the frequency of individuals of various ages and sizes during the whole year, suggests

it has developed tactics that allow its survival in this environment with small sizes (average 1.7 cm), which

characterizes it as an r-strategist. Anthropogenic pressures in areas where L. bolivianus lives in Brazil, as in

the area of the present study, make the species vulnerable because they increase its exposure to predators,

reduce its breeding sites, and increase its mortality from agricultural pesticides.

Keywords. Amapa, Amphibia, Brazil, Eastern Amazon, population structure

Citation: Sa-Oliveira JC, Costa-Campos CE, Araujo AS, Ferrari SF. 2020. Patterns of growth and natural mortality in Lysapsus bolivianus (Anura,

Hylidae, Pseudae) in an environmental protection area in the estuary of the Amazon River. Amphibian & Reptile Conservation 14(1) [General Section]:

156-162 (e229).

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: 10 July 2018; Accepted: 2 September 2019; Published: 6 April 2020

Introduction (Measey et al. 2016) or other poikilothermic vertebrates,

such as fishes and reptiles (Kozlowski and Teriokhin

While amphibian research has advanced considerably in

recent decades throughout the world, the ecology of most

species 1s still only poorly understood. This 1s reflected,

for example, in the large number of amphibian species

classified as Data Deficient by the IUCN (2016). The

comprehensive Global Amphibian Assessment (GAA)

estimated that more than 30% of the 7,881 recognized

amphibian species are currently threatened with extinc-

tion, and that several hundred may already be extinct

(Barnovsky et al. 2011; IUCN 2016).

The population dynamics of amphibians is still poorly

known in comparison with the diversity of this group

Correspondence. *ju/iosa@unifap.br

Amphib. Reptile Conserv.

1999; Shine 2010; Camargo et al. 2010; Loyola et al.

2008). This lack of data is a major obstacle to the devel-

opment of effective conservation measures (Stuart et al.

2004).

The life history strategies of a species are reflected in

a characteristic set of biological and demographic traits,

such as age at first sexual maturation, fertility and mor-

tality rates, reproductive patterns, and social organiza-

tion (Steams 1992; Ricklefs 1977). Growth and mortality

rates may be especially important for the understanding

of population structure and dynamics, and the capac-

ity of a species to cope with environmental disturbance

April 2020 | Volume 14 | Number 1 | e229

Sa-Oliveira et al.

(Radtke and Hourigan 1990; Pauly 1998).

In most tropical organisms, the assessment of growth

in rigid structures, such as bones, scales, and woody

stems, is impeded by the relative stability of the climate

and associated ecological variables (Boujard et al. 1991;

Marangoni et al. 2009). In this case, growth parameters

may be estimated indirectly through data on parameters

such as body size, which are more easily obtained from

wild populations. These data can provide insights into

the typical body sizes of different age groups, and the

definition of cohorts (Basson et al. 1988).

A number of non-linear models have been proposed to

describe growth patterns in animals, such as the Brody,

von Bertalanffy, and Gompertz functions, and logistic

procedures. The Bertalanffy model is the most popular

model for analyzing animal population dynamics. It is

based on the assumption that growth can be estimated

from the difference between the anabolic and catabolic

rates of an animal (Bertalanffy 1957; Hota 1994).

The frog Lysapsus bolivianus Gallardo, 1961 is a

semiaquatic anuran with both nocturnal and diurnal hab-

its, which inhabits the water surface in patches of float-

ing aquatic vegetation (Bosch et al. 1996; Garda et al.

2010; Santana et al. 2013). This species is small in size

(mean SVL = 17.6 mm), and is widely distributed in the

Amazon basin, ranging from the mouth of the Amazon

River in Brazil to northern Bolivia (Frost 2018). It is

found in the Rio Curiau Environmental Protection Area

(Rio Curiaii EPA) in Amapa state, northern Brazil, the

location of the present study (Melo-Furtado et al. 2014).

This protected area has been extensively impacted by hu-

man activities, such as deforestation, unregulated fishing,

construction of buildings, the accumulation of domestic

refuse, landfill of floodplains, and the indiscriminate use

of agricultural pesticides in the surrounding areas. All

these processes may impact the local biota, especially the

anurans, such as L. bolivianus.

The present study evaluated the growth and mortal-

ity parameters of the L. bolivianus population of the Rio

Curia EPA, together with estimates of longevity and

growth performance. These data will hopefully contrib-

ute to the development of effective conservation strate-

gies for the study species and other amphibians, as well

as the study area in general.

Materials and Methods

Study Area

The present study was conducted in the Rio Cu-

riau. Environmental Protection Area (00°09'00.7°N,

51°02'18.5”W), or Rio Curia EPA, which lies to the

north of the Amazon River estuary 1n the state of Amapa,

northern Brazil. The Rio Curiau EPA encompasses

21,700 ha, an area dominated by aquatic systems, such as

rivers and seasonal lakes. The local vegetation 1s main-

ly Cerrado savanna and floodplain forest. The region’s

Amphib. Reptile Conserv.

climate is humid equatorial (Am) in the Koppen-Geiger

classification system, with a mean monthly temperature

of 27.6 °C (range: 25.8—29 °C) and mean annual rainfall

of approximately 2,850 mm, with a monsoon period be-

tween February and May when the monthly precipitation

is around 400 mm (Alvares et al. 2013). The number and

sizes of the ponds found within the study area decrease

considerably during the dry season.

Sample Collection

Between January and December 2015, frogs were cap-

tured randomly by hand during the night, using 9 V flash-

lights. Frogs were collected by active searches along five

1 km transects in floating vegetation (Nymphoides indica

[L.] Kuntze and Salvinia auriculata Aubl.). The transects

were separated by a distance of at least 50 m and were

walked by three researchers during each survey (Crump

and Scott 1994). The frogs captured were examined to

determine their sex and age (adult or juvenile), based on

the presence of the nuptial sac in males and the distended

or flaccid abdomen (before or after spawning) of the ma-

ture females, and their positions in amplexus.

The snout-vent length (SVL, in mm) of each speci-

men was measured using a tape measure and calipers,

and the weight (Wz, in g) was recorded using a spring

balance (0.01 g precision). Sample collection was autho-

rized by the Brazilian Environment Institute (BAMA)

and the Information and Authorization System (SISBIO)

of the Chico Mendes Institute for Biodiversity Conser-

vation (ICMBio) through license number 34238-1. After

measurements were taken, all individuals were released

at the capture site.

Statistical Analyses

Deviations in the sex ratio were evaluated using a Chi-

square test with Yeats’ correction. The SVL values were

grouped into classes to permit the visualization of the

differences between adults and juveniles, and between

mature males and females. The difference in the mean

body size (SVL) between males and females was ana-

lyzed using a ¢ test. For this, the assumptions of normal-

ity and homoscedasticity were tested a priori using the

Kolmogorov-Smirnov and Bartlet tests, respectively. A

significance level of 5% was considered in all cases.

The total length-weight relationship was determined

by the Sparre et al. (1989) allometric equation Wt = a*L?,

where Wt = body weight (g), L = SVL (mm), and ‘a’ and

‘b’ = regression constants. Growth parameters were based

on the Von Bertalanffy equation, SVL=SVL, x (I- e **)

[Sparre and Venema 1998], where SVL = total snout-vent

length (mm) at age ¢, SVL, = asymptotic snout-vent length

(mm), K = growth rate (year'), ¢ = the age in years, and £,

= the nominal age at metamorphosis, assumed to be zero.

The constants K and SVL, were estimated by the Ford-

Walford model (Ford 1933; Walford 1946).

April 2020 | Volume 14 | Number 1 | e229

Growth and mortality of Lysapsus bolivianus in Brazil

3 40

o 30 @ Males

a m Females

wo 20

oc 10

13.2 to

14.6

14.6 to

16.0

16.0to 17.4to

17.4 18.8 20.2

Snout-vent length (mm)

18.8to 20.2to 21.6to 23.0to

21.6

(o>)

@ Juveniles

Relative frequency (%)

(jo)

11.6 to 13.0 to 14.4 to 15.8 to

13.00 144 15.8 17.2

Snout-vent length (mm)

23.0 24.4

Fig. 1. Relative frequency of the body size classes (SVL, snout—vent length; mm) recorded in the (A) adult males and females and

(B) juveniles of the Lysapsus bolivianus population from the Rio Curia EPA on the estuary of the Amazon River, in northern Brazil.

Longevity (4,,, or t,.) was calculated using the Tay-

lor (1958) equation, 4,,, = t, + (2.996/K), and natural

mortality (/) was estimated using the Hoenig (1983)

equation which is based on the empirical relationship ob-

served between M and the maximum age described by

the equation LnM = 1.46-1.01[Ln (t,_)], where t= the

maximum age in the population, and / = the natural rate

of mortality. The asymptotic weight (W_) was estimated

by converting L, to the corresponding weight using the

Pauly (1998) formula for the length-weight relationship

(W._, = a* SVL,’). The growth performance (@’) was esti-

mated by the Pauly and Munro (1984) formula: 0'= log

k + 2 log SVL... Juveniles were analyzed separately from

the adults due to their much faster juvenile growth rates

(mean = 16.0 + 3.6 days). As the sex of the juveniles

could not be determined, the data were pooled for this

age class.

Results

A total of 308 mature ZL. bolivianus individuals were ex-

amined, together with 71 individuals classified as juve-

niles. Overall, mature males (7 = 188) were significantly

more abundant than mature females, with nm = 120 (XY?

[with Yates’ correction] = 14.57; df= 1; p=0.001). How-

ever, the females were significantly larger, on average,

than males, with a mean SVL in the females of 19.81

+ 1.35 mm (range = 16.40—23.48 mm) versus 17.60 +

1.13 mm (14.11—20.17 mm) in the males (¢ = 15.26; df

= 306.0; p < 0.0001). Clear peaks in body size were ob-

served in both sexes (Fig. 2), with 79.0% of the adult

males having an SVL of 16.0-18.8 mm, and 78.2% of

the females at 18.8—21.6 mm. The juveniles presented

a mean SVL of 15.42 + 1.07 mm (range: 12.00-16.96

mm), with 81.13% of the specimens lying between 14.4

and 17.2 mm (Fig. 1).

Highly significant coefficients of determination were

recorded for the total length-weight relationships in both

adult males (R* = 0.63; F, ,.,. = 67.29; p < 0.0001) and

females (R? = 0.74; Foun = 521.51; p < 0.0001), indicat-

ing different models for the two sexes. The 5 values of

males and females were both lower than 3, which indi-

cate negative allometric growth (Fig. 2).

The relationship between the mean SVL at age t and

t+ 1 (SVL + 1) was described adequately by Walford’s

equation: SVL,,, = 12.971 + 0.4192 SVL, R* = 0.989 for

males and SVL,,,= 12.149 + 0.5198 SVL, R* = 0.959 for

females (Fig. 4). The intersection between the function

and the diagonal drawn through the origin provides the

value of L,, which was 21.20 mm in males and 25.47 mm

in females. Based on the formula L,~ L_. /0.95 (Taylor

1958), the estimated values of SVL, were 21.17 mm for

males, and 24.65 mm for females, values which are very

close to those derived from the graphs (Fig. 3). Estimated

growth parameters are presented in Table 1. In general,

the parameters were similar between males and females,

Table 1. Growth parameters for Lysapsus bolivianus specimens from the Rio Curiau EPA in Amapa, Brazil. SVL, = maximum length;

SVL,, = asymptotic length; k = growth constant; O' = growth performance; M = mortality; A,,. = longevity; n = sample size.

Parameter Males (n = 188)

K (year') 0.71

SVL, (mm) 21.20

Q' 2.50

Avs 4.20

SVL,,,, (mm) 20;17

0.64

Females (n = 120) Juveniles (n = 71)

0.70 0.81

25.60 59.50

2.65 3.45

4.28 3.70

23.48 16.96

0.65 12.86

M (year')

Amphib. Reptile Conserv.

158

April 2020 | Volume 14 | Number 1 | e229

Sa-Oliveira et al.

Weight (g)

10 12 14 16

Females

y= 0,0017529735

R*=0.74

n=120

Males

y= 0,0023K0 729

R? = 0.63

n=188

SVL (mm)

Fig. 2. Weight-length relationships in adult male and female Lysapsus bolivianus from the Rio Curiau EPA on the estuary of the

Amazon River, in northern Brazil.

although asymptotic size and growth performance varied

between the sexes. Growth rates, longevity, and natural

mortality were equal in the sexes.

Males grew 2.22 mm, on average, from an age of 3

to 6 months, 0.26 mm from 6 to 9 months, and only 0.04

mm from 9 to 12 months (Table 2). In females, growth

over these same intervals was 2.80 mm, 0.30 mm, and

0.10 mm, respectively. This variation was shown in the

Bertalanffy growth curves (Fig. 4), which followed dis-

tinct patterns in the males and females.

Discussion

The predominance of males recorded in the present study

was consistent with the findings of Melo-Furtado et al.

(2014). The male-biased sex ratio in L. bolivianus may

be advantageous for the fertilization of the largest pos-

sible number of eggs. Other studies have related devia-

tions in sex ratio to factors including differential growth

and mortality, as well as fluctuations in the availability

of nutrients and behavioral variations, all of which may

have varying influences on the proportion of the sexes

Table 2. Average snout-vent length at different ages calculated

for adult male and female Lysapsus bolivianus from the Rio

Curia EPA in Amapa, Brazil.

Average SLV (mm) at age indicated

3months 6 months 9 months 12 months

Male 18.68 20.90 21.16 21.20

Female 22.40 25.20 25.50 25.60

Amphib. Reptile Conserv.

at different stages of development (Hamilton and Zuk

1982: Vazzoler 1996; Kraab and Pen 2002; Fawcett et

al. 2011; Booksmythe et al. 2013).

The growth rates of both male and female L. bolivi-

anus were relatively high (> 0.5), which is typical of

species found in highly seasonal habitats, such as that of

the study area, as well as those that suffer high rates of

predation (Lowe-McConnell 1999). These species grow

rapidly, reaching maturity sooner with smaller asymp-

totic body lengths than larger species with slower growth

rates (Pauly 1998).

A reduced asymptotic length, while determined ge-

netically, may also be influenced by variables such as

the food supply and population density (Parker 1983;

Hubbell and Johnson 1987). Pauly (1998) concluded that

low values are typical of tropical species and may be at-

tributed to the combination of a number of factors, par-

ticularly temperature, given that higher temperatures ac-

celerate growth and metabolic rates but tend to decrease

the asymptotic size (Lomolino and Perault 2007). Faster

growth to maturity may also be a strategy to compensate

for predation pressure (Reznick et al. 1996).

The natural mortality rates in both sexes were moder-

ate to high, and varied proportionately with growth rates,

indicating that the principal causes of mortality in this

Species are predation and longevity, related to its rapid

life cycle (Keiber 1932; Pauly 1998). While the L. bolivi-

anus males grow faster than the females, both sexes have

similar longevities, which favors reproductive success in

a highly seasonal environment that is characterized by

long periods of drought.

April 2020 | Volume 14 | Number 1 | e229

Growth and mortality of Lysapsus bolivianus in Brazil

oe in 30

A: Males ww Bifemales. oo Jee

25 a x emales ;

a a SVL,, = 25.60

_ 20 ose SVL = 21.20 SAS 20 ee

E ae . E a

Sears - £ 45 F

se |) (i oie eee EE.

Stet as a (0

a >

n " Zz ;

° a 5 “

Ga 4

0 5 10 15 20 25 30 0 5 10 15 20 25 30

SVL (mm) SVL (mm)

Fig. 3. Plot of the Ford-Walford estimates of growth parameters (SVL, k) of adult (A) male and (B) female Lysapsus bolivianus

from the Rio Curia EPA in Amapa, Brazil. The values were estimated by the linear regressions between SVL and SVL+1 for each

gender, as SVL, = (a/I-b) and K = -log, b.

28 28

Females

c— = SVL = 25.6 [1-exp(-0.70t

oe Males =

S 16 SVL = 21.2 [1-exp(-0.711)] — 16

n >

12 12

8 8

12 3 4 5 6 7 8 9 10 11 12 13 12 3 4 5 6 7 8 9 10 11 12 13

Age (months) Age (months)

Fig. 4. Von Bertalanffy’s growth curves for (A) male and (B) female Lysapsus bolivianus from the Rio Curia EPA in Amapa, Brazil.

Conclusions Barnosky AD, Matzkel N, Tomiya S, Wogan GOU,

Swartz B, Quental TB, Marshall C, McGuire JL, Lind-

All parameters analyzed indicate an r type of life history sey EL, Maguire KC, et al. 2011. Has the Earth’s sixth

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Amphib. Reptile Conserv.

141-147.

Julio C. Sa-Oliveira is a biologist, with a Doctorate in Aquatic Ecology and Fisheries, and is cur-

rently a teacher at the Federal University of Amapa-Brazil, Julio has experience in the area of the

ecology of aquatic environments, with emphasis on bioecology, water quality assessment, and mod-

eling of ecosystems, populations, and communities.

Carlos E. Costa-Campos has a Ph.D. in Psychobiology from the Federal University of Rio Grande

do Norte, Brazil. Carlos has experience in the area of zoology (with an emphasis on amphibians

and reptiles), and is active in research on the natural history, ecology, behavior, and conservation of

herpetofauna. Currently he is a teacher at the Federal University of Amapa, Brazil.

Andrea S. Araujo has a Ph.D. in Psychobiology from the Federal University of Rio Grande do

Norte, Brazil, and is an Adjunct Professor HI of the Federal University of Amapa, also in Brazil.

Andrea has experience in zoology, with an emphasis on vertebrate zoology, working mainly on the

behavior, ethnozoology, and ecology of vertebrates.

Stephen F. Ferrari has a bachelor's degree (University of Durham, 1983) and Ph.D. (University

of London, 1988) in Biological Anthropology. Stephen is currently an Associate Professor I at the

Federal University of Sergipe, Brazil. He is also an ad-hoc consultant for CAPES and ICMBio, and

a member of the IUCN Primate Specialist Group. Stephen has experience in ecology, with emphasis

on primatology, working mainly in ecology, conservation, animal behavior, habitat fragmentation,

and environmental education.

162 April 2020 | Volume 14 | Number 1 | e229

Official journal website:

amphibian-reptile-conservation.org

Amphibian & Reptile Conservation

14(1) [General Section]: 163-173 (e230).

Persistence of snake carcasses on roads and its potential

effect on estimating roadkills in a megadiverse country

Laura Ximena Cabrera-Casas, Lina Marcela Robayo-Palacio, and *Fernando Vargas-Salinas

Grupo de investigacion en Evolucion, Ecologia y Conservacioén EECO, Programa de Biologia, Facultad de Ciencias Basicas y Tecnologias,

Universidad del Quindio, Armenia, COLOMBIA

Abstract.—The persistence of fauna carcasses on roads has been considered one of the most relevant factors

influencing estimates of road mortality in different taxonomic groups. However, there is a lack of information

in this regard in most Neotropical countries. The aim of the present research is to describe and quantify the

persistence of snake carcasses on two Colombian roads and its potential relationships with animal body size and

the frequency of vehicular traffic. Additionally, to illustrate the importance of correcting roadkill rate estimates

for carcass persistence, the roadkill rate of snakes on a secondary road in the study area is recalculated

using the results of carcass persistence time, instead of a previously selected arbitrary value (i.e., 7 days).

To estimate the carcass persistence time, eighty-one snake carcasses of diverse body sizes (mean length =

46.90 cm + (SD) 38.46, range = 10.1-—224 cm) were placed over sampling points distributed equally on two roads

with different traffic frequencies (a primary road with > 2,500 vehicles/day and a secondary road with < 1,000

vehicles/day). Snake carcass degradation was monitored until their disappearance from the road surface. The

median persistence time of carcasses on roads was 7.16 h (5.20 h and 14.16 h on the primary and secondary

roads, respectively). According to field experiments, around 75% of the carcasses can disappear from the road

surface within 30 h after a snake has been killed. The principal cause of the disappearance of carcasses during

the day was degradation due to vehicular traffic. As the carcasses tended to increase in size, the difference

in their persistence between types of roads increased, with lower persistence on the primary road than on

the secondary road. The conclusion from these observations is that excluding carcass persistence time from

snake roadkill rate calculations leads to high underestimates of mortality. Therefore, the problem of snakes that

are roadkill on the road infrastructure in Colombia is much greater than previously considered.

Keywords. Andes, Colombia, mortality, Reptilia, road ecology, Serpentes, Squamata, vehicular traffic

Citation: Cabrera-Casas LX, Robayo-Palacio LM, Vargas-Salinas F. 2020. Persistence of snake carcasses on roads and its potential effect on

estimating roadkills in a megadiverse country. Amphibian & Reptile Conservation 14(1): 163-173 (e230).

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: 6 August 2018; Accepted: 8 December 2019; Published: 14 April 2020

Introduction

Among the ecological effects attributed to the presence

of roads is animal mortality due to collision with

vehicles (Forman et al. 2003; van der Ree et al. 2015).

Fauna roadkill rates have been estimated in thousands

to millions of animals per year (Huiyjser and McGowen

2010; Marsh and Jaeger 2015). For example, in Australia,

it has been estimated that up to 12 million vertebrates

are killed yearly on roads, while in the United States

the estimates are 1 million vertebrates per day (Bennett

1991; Forman and Alexander 1998). However, those

roadkill rates can be severely underestimated (Loss et al.

2014; Ruiz-Capillas et al. 2015; Santos et al. 2016) given

methodological biases and that many carcasses disappear

before being registered by researchers due to factors such

as rain, wind, scavenging activity, or vehicular traffic

(Bafaluy 2000; Ratton et al. 2014; Slater 2002; Taylor

and Goldingay 2004). In fact, roadkill estimates are

generally evaluated based upon time ranges of carcass

persistence on roads that have been arbitrarily selected or

with conservative approximations (Kl6écker et al. 2006).

In Latin America, several studies have documented

wildlife roadkill rates (e.g., Novelli et al. 1988; Pinowsk1

2005; Delgado 2007; Coelho et al. 2008; Barri 2010;

Royas-Chacon 2010; Vargas-Salinas et al. 2011;

Contreras-Moreno et al. 2013; De La Ossa-Nadjar and

De La Ossa 2013; Seijas et al. 2013). Nevertheless,

few studies have tested the factors affecting carcass

persistence on the roads and hence, have not quantified

the discrepancy between the number of carcasses counted

by researches and the real number of animals killed on

the roads (Santos et al. 2011; Santos et al. 2016; Teixeira

et al. 2013). This is concerning because the accurate

Correspondence. cabreracasaslx@gmail.com (LXC), Imrobayo.palacio@gmail.com (LMR), *fvargas@uniquindio.edu.co (FVS)

Amphib. Reptile Conserv.

April 2020 | Volume 14 | Number 1 | e230

Roadkill snake persistence in Colombia

75°40'W

75°39'W

4°42'N

COLOMBIA

4°41'N

“t

4°40'N

75°38'W

: Canon del Rio‘Barbas. ee

BM a ye road."

Reserva Bremen es 4

75°37'W 75°36'W

¢ *

“aah

. 4

Primary” *) |

Fig. 1. Geographic location of the study area. Primary road (Autopista del Café) and the secondary road connecting Autopista

del Café with the town of Filandia in the department of Quindio, Central Andes of Colombia. Adapted from SIG Quindio 2016.

http://190.85.164.56/sigquindioiii/

quantification of roadkill rates is crucial for assessing

the negative impact of roadkills in animal populations,

and for optimizing the allocation of the limited economic

resources for management and conservation (Slater

2002; Fahrig and Rytwinski 2009).

Snakes are one of the most commonly killed

vertebrates on roads, which has been attributed to several

non-exclusive factors (Andrews et al. 2008). For instance,

snakes may actively seek out roads for thermoregulation

(Rosen and Lowe 1994; Ashley and Robinson 1996) and

they are especially vulnerable to being killed because they

tend to freeze when facing an anthropogenic stimulus

such as the artificial light of approaching vehicles

(Andrews and Gibbons 2005). In addition, many snakes

are killed intentionally by drivers because of erroneous

preconceptions of their venomous nature (Secco et al.

2014). Given that many species of snakes exhibit a small

body size, their carcasses are expected to disappear from

the road in less than one or two days, a shorter period than

some protocols for monitoring roadkill (Antworth et al.

2005; Langen et al. 2007; DeGregorio et al. 2011; Santos

et al. 2011). Therefore, the number of road-killed snakes

can be highly underestimated if carcass persistence is not

factored into roadkill rate calculations.

The only national estimate of snake roadkill for

Colombia was published by Lynch (2012), based on

records obtained on a road in Brazil (Monteiro et al. 2011)

and a road in the western Andes of Colombia (Vargas-

Salinas et al. 2011). That author mentions that mortality

in Colombia due to collisions with vehicles could range

from 52,600 to 176,660 snakes/yr in the primary road

network of the country (10,300 km). However, that

estimate is inaccurate because roadkill rates of snakes

Amphib. Reptile Conserv.

vary according to the characteristics of the roads, as well

as features associated with the ecosystem bisected by

the roads (Forman et al. 2003; van der Ree et al. 2015).

Furthermore, estimates of roadkill rates provided by

Lynch (2012) for Colombia are based on a conservative

approximation of carcass persistence on roads (e.g., 3.5

days; Vargas-Salinas et al. 2011).

In this study, an initial estimate was made of the time

that a snake carcass can persist on two roads in Colombia.

Then, the relationships between the carcass persistence

time and both snake body size and traffic frequency

were examined. Finally, the roadkill rate of snakes on

a secondary road in the Central Andes of Colombia

studied by Quintero-Angel et al. (2012) was recalculated

by taking into account the results obtained here about

carcass persistence. This recalculation was made to

illustrate the importance of correcting roadkill rates for

carcass persistence, and also to improve knowledge of

road ecology, specifically, wildlife roadkills in Colombia.

With these analyses, we seek to draw attention to the

ecological impacts of roads on this country's snake fauna.

Materials and Methods

Study Area and Methodology

This study was conducted in a 4.5-km stretch along

two roads located in the department of Quindio, in the

Central Andes of Colombia (Fig. 1). One is a secondary

road that connects the Autopista del Café (a major

highway system) with the town of Filandia, which has an

average width of 8 m and a traffic frequency of <1,000

vehicles/day. The other is a primary road (Autopista del

April 2020 | Volume 14 | Number 1 | e230

Cabrera-Casas et al.

Fig. 2. Images of some species of snakes present in the study area and used in the field experiments. (A) Mastigodryas boddaerti,

(B) Tantilla melanocephala, (C) Dipsas sanctijoannis, (D) Erythrolamprus epinephelus, (E) Leptodeira annulata, (F) Oxyrophus

petolarius. Photos by Lina M. Robayo-Palacio (A), Fernando Vargas-Salinas (B—D), Ana Maria Ospina-L (E), and Wolfgang

Buitrago-Gonzadlez (F).

Café) with a traffic frequency of >2,500 vehicles/day.

This road is bordered by two margins, each with a width

of 8 m. Because the two margins were separated by a

median strip, and the field experiments were performed

in just one of the margins (see below), the carcasses were

assumed to be subjected to the impact of half of the traffic

level (i.e., ~1,250 vehicles/day). The minimum distance

between the two 4.5-km road segments was 1.5 km; and

since the road stretches are close to each other, climatic

conditions were assumed to be similar between them.

The temperature in the study area varies between 12

and 18 °C, with 83% mean annual relative humidity,

and a mean precipitation of 2,515 mm/yr. The rainfall

regime in this area is bimodal, with high precipitation

levels during April-May and October-November (CRQ

2010); however, during the sampling period (August

2015—March 2016) summer environmental conditions

predominated due to the “El Nifio” phenomenon (NOAA

Amphib. Reptile Conserv.

http://www.noaa.gov/understanding-el-nino). The

landscape surrounding both roads consisted of paddocks,

croplands, and some remnants of bamboo (Guadua

angustifolia) and secondary forest (Serna 2012).

Between August 2015 and March 2016, 81 carcasses

of snakes were placed randomly in the two roads under

study. Nine snake carcasses were placed on each road

spaced 500 m apart. The carcasses were placed on the

roads between 0630 and 0700 h, and were monitored

continuously throughout the day until 1800 h. The

monitoring consisted ofdirect observations with binoculars

at a distance of 50 m to avoid interfering with potential

scavenger activity. The time of carcasses persistence and

the causes of their eventual disappearance were recorded.

A carcass was considered to be no longer detected when

it was absent from the asphalt and the ground next to the

road, or when the body was fragmented and degraded

to a level that became unrecognizable from a potential

April 2020 | Volume 14 | Number 1 | e230

Roadkill snake persistence in Colombia

Ln Snake body weight (g)

-2

2 3 4 5 6

Ln Snake body length (cm)

Persistence of carcass on road (h)

i=)

150

100

Primary road @

0 Secondary roadO

0 50 100 150 200

Snake body length (cm)

Fig. 3. (A) Relationship between weight and body length (Ln = natural logarithm) of snake carcasses used in this study. (B)

Relationship between body length of snake carcasses and their persistence time on two roads with different levels of vehicular

traffic.

observer located at the margin of the road. When

carcasses disappeared during night (due to scavengers

or any other factors), they were assumed to be removed

the following morning when the monitoring restarted; in

this way we were conservative in our estimates. Snake

carcasses used in the present study were obtained from

deaths and roadkills collected between April 2015 and

March 2016 (Fig. 2; Appendix 1) in various places in the

department of Quindio, Colombia. This study only used

snakes recently run over with a non-flattened body and

with fresh tissues and blood.

Although the carcasses used were taken from different

locations in the department of Quindio, the species were

locally distributed in the area of monitoring. The carcasses

were frozen until the beginning of the field experiment.

Individual taxonomic identification of the road-killed

snakes was conducted based on prior knowledge of the

species (Quintero-Angel et al. 2012; Vanegas-Guerrero

et al. 2016) and taxonomic descriptions in the literature

(e.g., Lynch and Passos 2010; Peters and Orejas-Miranda

1970).

Data Analysis

A t-test was used to verify that there were no differences

in the total length (hereafter, body size) or mass of the

carcasses used in the field experiments between the

primary and the secondary roads. A high correlation

between the body size and weight of the snake carcasses

was corroborated using a linear regression analysis;

therefore, only the body size of the individual was used

for subsequent analysis.

To achieve the first aim of this study, the median

value of carcass time persistence for each type of road

was estimated. In addition, a Kaplan-Meier estimator

Amphib. Reptile Conserv.

(Therneau and Grambsch 2000) was performed to

calculate the probability of a carcass remaining on the

road surface (1.e., potentially detectable by a researcher)

through time. These probabilities were obtained for the

primary and secondary roads separately. For the second

objective, an analysis of covariance (ANCOVA) was

applied; where the snake body size and type of roadway

(primary, secondary) were the explanatory variables,

while the carcass persistence time was the response

variable. To achieve the third objective, the median

persistence time of snake carcasses obtained for the

secondary road in the study was used to recalculate the

roadkill rate published by Quintero-Angel et al. (2012).

This estimation was reasonable because that study was

made in the same secondary road stretch as the current

experiments. The new roadkill rate estimate (RRE) was

made using the following equation:

Time between

Number of Days of year samplings

carcass (365) (days)

RRE=

Length of surveyed Monitoring Median persistence

road stretch time period time (in days)

(Km) (days) of carcass

Results

There were no differences in the body size and weight of

the carcasses between the primary and secondary roads

(body size: t = 0.283, df = 79, P = 0.77; body weight:

t = 0.451, df = 79, P = 0.653), and there was a strong

positive relationship between the body size and weight

of the snake carcasses (R? = 0.73, F = 225.135, 6 = 0.86,

P < 0.001; Fig. 3A).

In this study, 81 carcasses were used which belonged

to 14 species distributed in three families: Colubridae

(41 individuals), Dipsadidae (39), and Elapidae (1).

April 2020 | Volume 14 | Number 1 | e230

Cabrera-Casas et al.

Survival probability

Secondary road

Primary road

0 30 60 90 120 150

Time (hours)

180 210

Fig. 4. Survival curves (1.e., probability of persistence on the

road) for snake carcasses on primary and secondary roads in the

municipality of Filandia, department of Quindio, Central Andes

of Colombia. Kaplan-Meier estimates based on a sample size N

= 81 snake carcasses.

The median persistence time of the snake carcasses on

the primary road was 5.20 h (range = 0.26—145.0 h; N

= 41 individuals), while on the secondary road it was

14.16 h (range = 0.27—193.90 h; N = 40 individuals).

By combining the data from both types of roadway, the

median persistence time of the carcasses was 7.16 h (range

= 0.26—-193.90 h). The persistence probability dropped

substantially beyond 24-48 h for both the primary and

secondary roads; in other words, after a snake has been

killed, the probability that a researcher would be able

to record the carcass on the road ts less than 0.25 after,

for example, 30 h (Fig. 4). During the daytime, the main

cause of carcass disappearance was degradation due to

vehicular traffic (65 of the 81 carcasses), and only three

carcasses were removed by scavengers (birds). Thirteen

other carcasses disappeared at night so it was not possible

to determine the cause (see Appendix 1).

Compared to carcasses with a smaller body size,

snakes with a larger body size tended to persist longer

on the roads (ANCOVA F = 169.68, df = 1, P < 0.001).

Although the type of road did not influence the carcass

persistence time (F = 0.124, df = 1, P = 0.725), the

analysis of covariance revealed that carcass size/type

of road interaction had a significant effect on carcass

persistence (F = 11.35, df = 1, P = 0.001). If the carcass

was small, it tended to persist for the same time on the

primary road as on the secondary road; however, when

the carcass was large, the difference in persistence

time between road types increased, being lower on the

primary road compared to the secondary road (Fig. 3B).

Using these results of median carcass persistence time

on the secondary road, the roadkill rates of snakes for

the location reported by Quintero-Angel et al. (2012),

Amphib. Reptile Conserv.

increased substantially, from 78.8 individuals/km/year to

934.83 individuals/km/year.

Discussion

The estimate of snake carcass persistence time on roads

followed a general trend found in similar studies. In

North America, South America, and Europe, nearly 50%

of carcass disappearance from the roadway occurs during

the first 8 to 24 h (DeGregorio et al. 2011; Santos et al.

2011).

Snake carcasses with a larger body size were found

to persist longer than carcasses with a smaller body size

on the roads. This result is in agreement with Santos et

al. (2011), but disagree with the findings of Antworth et

al. (2005), DeGregorio et al. (2011), and Hubbard and

Chalfoun (2012). The results here differ from those

observed by Antworth et al. (2005) possibly because a

relatively small range of snake carcass sizes (54.3—136

cm) was used in that study compared with the current

study (range: 10.1—224 cm; see Appendix 1). Regarding

the results obtained by DeGregorio et al. (2011), these

authors placed the carcasses on the roadsides to evaluate

only the effect of scavengers on carcass persistence,

excluding the effect of traffic. In this study, carcass

persistence was mainly determined by the action of

vehicular traffic and not by scavengers. Finally, it is

possible that Hubbard and Chalfoun (2012) did not find

a relationship between carcass body size and persistence

time because they used portions of fishes for their field

experiments, which do not resemble the consistency of

snake carcasses that are covered with scales and include

a bone structure (1.e., spinal column). Characteristics

such as scales, the presence or absence of hair, and spines

on animal bodies have been found to allow corpses to

better withstand the passage of vehicles and last longer

on roadways (Santos et al. 2011).

The lack of statistical differences in the persistence

time of carcasses between the primary and secondary

roads might be attributed to the relatively small difference

in traffic that could potentially hit the carcasses (~1,250

vehicles/day vs. <1,000 vehicles/day, respectively).

Nevertheless, Santos et al. (2011) also found no

significant differences on snake carcass persistence

among four types of roadways (<1,000; 1,000—4,000;

4,000—10,000; and >10,000 vehicles/day). However, this

result could be a sampling artifact, given the relatively

small sizes of the snakes used in that study (15-240

g), which may not have allowed the recording of an

interaction between the level of traffic and the animal’s

body size. The present study used snake carcasses varying

between 0.3 and 1,147.29 g (see Appendix 1) and found

that differences in the persistence of carcasses among

roads with a different frequency of traffic emerged with

particularly large animals. A similar situation may have

occurred in Ratton et al. (2014), where a slight variability

in weight (mean = 39.6 + (SD) 2.1 g) of poultry corpses

April 2020 | Volume 14 | Number 1 | e230

Roadkill snake persistence in Colombia

(Gallus domesticus) may have accounted for the absence

of significant differences in the persistence of carcasses

between a highway and an unpaved roadway with sparse

vehicular traffic.

The recalculation of the snake roadkill rate obtained

by Quintero-Angel et al. (2012) on a secondary road of

Central Andes of Colombia, pointed out that including the

data of carcass persistence time may increase estimates

of snake roadkill significantly; specifically, an 11.86-fold

increase. However, the same level of increase should

not necessarily be found in other localities or with other

organisms. In this regard, recent studies testing previous

estimates of roadkills in countries such as Brazil, Spain,

United States of America, and Wales, showed a high

underestimation in roadkill values (Slater 2002; Loss et

al. 2014; Ruiz-Capillas et al. 2015; Santos et al. 2011;

Santos et al. 2016).

Because roadkill rate and carcass persistence time can

vary significantly between areas with different landscape

configurations, weather regimes, and roads, it was not

possible to estimate snake roadkill at the country level.

The simple extrapolation of the results from the current

study area (which is small in proportion to the size of

the road network in Colombia) is not adequate. This

is especially true for a country like Colombia, which

exhibits an intricate topography that is reflected in a

high diversity of ecosystems and a concomitant spatial

variation of snake diversity (Lynch 2012). Additional

studies examining snakes road-killed in different regions

and ecosystems of Colombia could help to provide a

better estimate of snake mortality for the country (e.g.,

Payan et al. 2013; Castillo-R et al. 2015; De La Ossa-V

and Galvan-Guevara 2015; Rincon-Aranguri et al.

2019); however, in those studies carcass persistence was

not evaluated.

Finally, according to the results obtained here, in

order to accurately estimate the roadkill rate for snakes,

and many other small vertebrates, a daily monitoring

schedule should be adequate (see Santos et al. 2011).

This monitoring frequency is higher than those reported

in previous studies from Colombia and other countries.

Nevertheless, it is important to remember that this

sampling capacity would depend to a large degree on the

economic support available.

Conclusion and Perspectives

This study highlights that the problem of road-killed

snakes on the road infrastructure of Colombia is greater

than previously believed. More data on carcass persistence

time obtained from different localities and different types

of roads (e.g., paved, non-paved) around the country are

necessary for determining an accurate estimate of the

number of snakes killed annually in Colombia. Given

that collisions may be related to a species population

size (Rosen and Lowe 1994; Row et al. 2007; Fahrig

and Rytwinski 2009), these results bear important

Amphib. Reptile Conserv.

implications for visualizing the need for conservation

plans against roadkills. Snakes are important prey and

predators in terrestrial ecosystems (Diller and Johnson

1988; Godley 1982), and for this reason, the impact of

roadways on snake populations might even be reflected

at the ecosystem level (Forman and Alexander 1998).

Acknowledgments.—The authors thank Julian Alberto

Rios-Soto, Alexander Marquez Cuervo, Cristian Gaviria

Londofio, Leidy Fernanda Daza Benavides, Oscar

Eduardo Grayjales Hernandez, Cristian Gonzalez-Acosta,

Maria Camila Basto-Riascos, Carlos Mario Gomez

Lopez, and Larry Alvarez Rodas for their help collecting

the carcasses. Gratitude is also expressed to Oscar Daniel

Medina Barrios, Julian Arango-Lozano, and Wolfgang

Buitrago-Gonzalez for confirming the taxonomic

identity of the snakes that were run over. We also thank

Universidad del Quindio for providing its infrastructure,

equipment, and facilities in the Bengala Farm for our

stay during field work. Ana Maria Ospina-L, Daniela

Layton, Carolina Lopez-Castafieda, Evelyn Zufiga,

Juan Carlos Gonzalez-Vélez, Dina Lucia Rivera-Robles,

Maria Paula Toro, and Juan Jaramillo Fayad improved

previous versions of this manuscript. Finally, thanks to

the journal editors and two anonymous referees for their

invaluable commentaries and suggestions.

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Laura Ximena Cabrera-Casas and Lina Marcela Robayo-Palacio are both graduates

from the Biology program at Universidad del Quindio, Colombia. Their research interests

are focused on studying inter-specific interactions and their effects on the structure and

composition of Neotropical reptile assemblages.

Fernando Vargas-Salinas is an Associate Professor in the Biology program at

Universidad del Quindio in Armenia, Colombia, where he is in charge of the academic

areas of ecology, animal behavior, and herpetology. Fernando’s research interests are

focused on the ecological and behavioral aspects of Neotropical vertebrates and their

relations with evolutionary processes and conservation biology. For this, he mainly

studies acoustic communication and reproductive behavior in anurans, and the diversity of

amphibians and reptiles in environments disturbed by the implementation of agricultural

activities or the presence of roads and urban complexes.

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Cabrera-Casas et al.

Appendix 1. Taxonomic identity and associated information of snake carcasses used in the field experiments.

Tae Collection Place of origin Body length Weight Cause of Period of

date (cm) (g) disappearance disappearance

COLUBRIDAE

Chironius monticola 7/10/2015 Filandia 125 179.5 traffic day

Chironius monticola 27/05/2015 Buenavista 48.9 13.8 traffic day

Dendrophidion bivittatus 27/11/2015 Filandia 20) 2.8 traffic day

Lampropeltis triangulum 4/10/2015 Filandia 69.3 93.33 traffic day

Lampropeltis triangulum 13/10/2015 Filandia 69 76 traffic day

Mastigodryas boddaerti 13/04/2015 Calarca 79.4 86.43 traffic day

Mastigodryas boddaerti 24/03/2016 Quimbaya 140 160.8 traffic day

Spilotes pullatus 4/11/2015 Quimbaya 224 1,147.29 traffic day

Tantilla melanocephala 12/08/2015 Filandia 26 2:6 traffic day

Tantilla melanocephala 26/06/2015 Filandia 29.4 5.16 traffic day

Tantilla melanocephala 10/05/2015 Filandia 43.1 6.4 traffic day

Tantilla melanocephala 20/08/2015 Filandia 39 Ded traffic day

Tantilla melanocephala 15/05/2015 Filandia 27 4.5 traffic day

Tantilla melanocephala 25/08/2015 Autopista del Café 12 0.61 traffic day

Tantilla melanocephala 20/08/2015 Autopista del Café 11.5 0.41 traffic day

Tantilla melanocephala 1/09/2015 Autopista del Café 38.2 6.17 traffic day

Tantilla melanocephala 19/09/2015 Filandia Dy 25 traffic day

Tantilla melanocephala 10/09/2015 Filandia 35.5 6.22 traffic day

Tantilla melanocephala 10/09/2015 Filandia 33 3.96 traffic day

Tantilla melanocephala 10/09/2015 Filandia 24 3.5 traffic day

Tantilla melanocephala 13/02/2016 Filandia 42.4 49 traffic day

Tantilla melanocephala 15/02/2016 Filandia 25 25 traffic day

Tantilla melanocephala 15/02/2016 Filandia 125 0.5 traffic day

Tantilla melanocephala 7/02/2016 Circasia 16 0.7 traffic day

Tantilla melanocephala 7/11/2015 —Autopista del Café D5 24 traffic day

Tantilla melanocephala 7/11/2015 Autopista del Café 28.5 3:1 unknown night

Tantilla melanocephala 7/11/2015 = Autopista del Café 10.7 0.5 traffic day

Tantilla melanocephala 7/11/2015 Autopista del Café 20 25 traffic day

Tantilla melanocephala 10/07/2015 Filandia 39,2 4.19 traffic day

Tantilla melanocephala 10/07/2015 Autopista del Café 10.5 0.35 traffic day

Tantilla melanocephala 5/09/2015 Filandia 30.33 6.6 traffic day

Tantilla melanocephala 5/09/2015 Filandia 25.4 3.87 traffic day

Tantilla melanocephala 2/09/2015 Autopista del Café 10.1 0.31 traffic day

Tantilla melanocephala 2/09/2015 Filandia 40.2 47 traffic day

Tantilla melanocephala 29/10/2015 Filandia 30.34 4.61 traffic day

Tantilla melanocephala 19/10/2015 Filandia 19 17 traffic day

Tantilla melanocephala 6/11/2015 Filandia 12 0.3 traffic day

Tantilla melanocephala 19/10/2015 Filandia | Wie 0.92 traffic day

Tantilla melanocephala 19/10/2015 Filandia 23.4 1.85 traffic day

Tantilla melanocephala 27/02/2016 Filandia 28 2.6 unknown night

Tantilla melanocephala 17/02/2016 Filandia 13.5 0.8 traffic day

Amphib. Reptile Conserv. 171 April 2020 | Volume 14 | Number 1 | e230

Roadkill snake persistence in Colombia

Appendix 1 (continued). Taxonomic identity and associated information of snake carcasses used in the field experiments.

Taxon

DIPSADIDAE

Atractus cf. melanogaster

Atractus sp.

Clelia sp.

Dipsas cf. sanctijoannis

Erythrolamprus

epinephelus

Erythrolamprus

epinephelus

Erythrolamprus

epinephelus

Erythrolamprus

epinephelus

Erythrolamprus

epinephelus

Erythrolamprus

epinephelus

Erythrolamprus

epinephelus

Erythrolamprus

epinephelus

Erythrolamprus

epinephelus

Amphib. Reptile Conserv.

Collection

date

20/08/2015

5/09/2015

15/11/2015

19/08/2015

20/01/2015

5/09/2015

30/02/2016

4/12/2015

7/03/2016

4/09/2015

30/08/2015

23/11/2015

8/11/2015

4/12/2015

17/03/2016

17/02/2016

20/09/2015

28/09/2015

22/07/2015

19/10/2015

11/10/2015

6/11/2015

12/02/2016

5/03/2016

10/09/2015

7/06/2015

21/05/2015

20/08/2015

25/09/2015

10/07/2015

2/09/2015

29/10/2015

7/11/2015

30/11/2015

Place of origin

Filandia

Body length

(cm)

38.8

Autopista del Café 30.6

Filandia

Quimbaya

Filandia

Circasia

Filandia

19:5

27.5

31.4

43.1

75.8

115.5

125.5

154.3

135

80.5

65.1

116.6

50.2

16.3

35.4

44.2

26.8

172

Weight

(g)

4.18

4.24

3.4

mal

Cause of

disappearance

traffic

unknown

traffic

unknown

traffic

unknown

traffic

unknown

traffic

scavenger

traffic

unknown

traffic

unknown

traffic

unknown

traffic

scavenger

traffic

unknown

Period of

disappearance

day

night

day

night

day

night

day

night

day

night

day

night

day

night

day

night

April 2020 | Volume 14 | Number 1 | e230

Cabrera-Casas et al.

Appendix 1 (continued). Taxonomic identity and associated information of snake carcasses used in the field experiments.

Collection nee Body length Weight Cause of Period of

Taxon Place of origin , *

date (cm) (g) disappearance disappearance

Erythrolamprus 27/12/2015 Filandia AO QF traffic day

epinephelus

Oxyrhopus petolarius 12/11/2015 Caicedonia 295 2.09 traffic day

Oxyrophus petolarius 29/10/2015 Filandia 595 24.44 scavenger day

Leptodeira annulata 7/03/2016 Armenia 31.2 N21 unknown night

ELAPIDAE

Micrurus mipartitus 4/04/2015 Quimbaya- DD, 1.12 traffic day

Filandia road

Amphib. Reptile Conserv. 173 April 2020 | Volume 14 | Number 1 | e230

Official journal website:

amphibian-reptile-conservation.org

Amphibian & Reptile Conservation

14(1) [General Section]: 174-182 (e231).

Effect of vegetation and abiotic factors on the abundance and

population structure of Crocodylus acutus (Cuvier, 1806) in

coastal lagoons of Colima, Mexico

'2.5Sergio Aguilar-Olguin, ?°Maria Cruz Rivera-Rodriguez, *Helios Hernandez-Hurtado,

‘Ricardo Gonzalez-Trujillo, and **Maria Magdalena Ramirez-Martinez

‘Universidad de Guadalajara, Centro Universitario de la Costa Sur. Av. Independencia Nacional 151,48900, Autlan de Navarro, Jalisco, MEXICO

2UMA Centro Ecologico de Cuyutlan “El Tortugario,”” Av. Lopez Mateos S/N, Poblado de Cuyutlan 28350, Armeria, Colima, MEXICO Universidad

de Guadalajara, Centro Universitario de la Costa, Av. Universidad 203, Ixtapa, 48280, Puerto Vallarta, Jalisco, MEXICO ‘Centro de Investigacion

para los Recursos Naturales, Antigua Normal Rural de Salaices, C. P. 33941, Lopez, Chihuahua, MEXICO ‘Universidad de Colima, Facultad de

Ciencias Biolégicas y Agropecuarias, Autopista Colima-Manzanillo Km 40, La Estacion, C. P. 28930, Tecoman, Colima, MEXICO

Abstract.—Crocodile populations are affected by their environment, and disturbance of that environment leads to

changes in their physiology and behavior. Using nocturnal spotlight counts, the influences of vegetation type and

several abiotic factors on populations of Crocodylus acutus were evaluated in Colima, Mexico. Six interconnected

lagoons with a known presence of crocodiles were selected, and the largest (Laguna de Cuyutlan) was divided

into four sections. Differences in crocodile density and size classes among these lagoons were determined, and

the effects of abiotic factors and vegetation type on the density and distribution of crocodiles were identified.

Salinity could influence the crocodile populations, since low crocodile densities were observed in lagoons with

high salinity. Average densities of crocodiles of 0.2—8.3 individuals/km and 0—5.9 ind/km were recorded during

the rainy and dry seasons, respectively. The average densities of crocodiles of size classes |, Il, and Ill ranged

from 0.3-1.7 ind/km, whereas those of size classes IV and V ranged from 0.1—1.8 ind/km. Population densities of

crocodiles were associated with factors such as salinity (<1%), and since the hatchlings and juveniles are the

most vulnerable to conditions of high salinity, they are drawn to sites of lower salinity, such as those with aquatic

and mangrove vegetation. This suggests that C. acutus can find refuge and food in the mangrove vegetation and

water at ambient temperatures of 3.9-6.3 °C. Variations observed in both the water and ambient temperatures

probably did not affect the normal thermoregulation processes of the crocodiles, since they can adopt a strategy

of thermoconformity in response to even minor variations in temperature. There were significant differences

(P <0.05) among the lagoons in terms of salinity, aquatic and mangrove vegetation, and water and ambient

temperatures. The coastal lagoons of Colima provided suitable habitats for crocodile distribution, but increased

salinity led to the movement of crocodiles towards areas supplied with freshwater.

Keywords. American Crocodile, depth, habitat, Pacific, salinity, wetlands

Citation: Aguilar-Olguin S, Rivera-Rodriguez MC, Hernandez-Hurtado H, Gonzalez-Trujillo R, Ramirez-Martinez MM. 2020. Effect of vegetation and

abiotic factors on the abundance and population structure of Crocodylus acutus (Cuvier, 1806) in coastal lagoons of Colima, Mexico. Amphibian &

Reptile Conservation 14(1) [General Section]: 174-182 (e231).

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: 24 October 2018; Accepted: 4 May 2019; Published: 18 April 2020

1987; Thorbjarnarson et al. 2006; Mazzotti et al. 2007;

Cupul-Magafia 2012). The Convention on International

Trade in Endangered Species of Wild Fauna and Flora

Introduction

The American Crocodile, Crocodylus acutus, has

the widest species distribution of its genus on the

American continent. It is found from southern Florida

to Colombia on the Atlantic side, and from Mexico to

Peru on the Pacific side of the continent (Brandt et al.

1995). The species inhabits shallow waters and, while it

prefers freshwater, the American Crocodile can tolerate

high salinities and use saltwater routes to move to

reproduction, feeding, nesting, and refuge areas (Lang

(CITES) lists C. acutus in Appendix I, the International

Union for the Conservation of Nature (IUCN) lists the

species as Vulnerable, and the Official Mexican Norm

059-SEMARNAT-2010 considers it a species subject to

special protection (DOF 2010).

Large specimens can be found at great depths in

Costa Rica, and high water salinity levels lead to a

greater dispersion of the animals (Mauger et al. 2012).

Correspondence. '*°zanyya@hotmail.com; ®> mary_cruz@live.com.mx; *hhh0474@hotmail.com; * ecologia.rgt@gmail.com;

ie 81a. TEs

'*maleni.ramirezm@gmail.com

Amphib. Reptile Conserv.

April 2020 | Volume 14 | Number 1 | e231

Aguilar-Olguin et al.

19°20°0°N

19°10'0"N

19°0'0"°N 19°10'0°N

19°0'0"°N

18°50'0"N

18°40°0°N

18°40'°0"°N

18°30'0°N

T T T

104°10°0"W 104°0°0"W 103°S0°O"W

104°20'0"W

Fig. 1. Selected sites in the study area. Acronym definitions and

characteristics of the sites are given in Table 1.

Relationships between low crocodile densities and

increased water temperature, high salinity, and poor

vegetation cover have been reported in Ecuador (Carvajal

et al. 2005). However, Cherkiss et al. (2011) did not find

a relationship between water or air temperature and

crocodile densities in Florida, although these authors

did observe that all crocodile size classes preferred

protected canals and ponds of low salinity surrounded by

vegetation. The distribution of American Crocodiles in

Colombia is influenced mainly by resource availability

(Balaguera and Gonzalez 2008). It is therefore important

to determine the relationships between these variables

and crocodile densities in Mexico, compared to other

populations of this species.

While Garcia-Grajales and Buenrostro-Silva (2014)

did not analyze salinity values on the coast of Oaxaca,

Mexico, these authors determined that high crocodile

densities were associated with mangrove vegetation

since the crocodiles preferred to stay hidden among

the mangrove roots. This finding coincides with

previous reports from Jalisco and Veracruz, where the

density of crocodiles C. acutus (Cupul-Magafia 2012)

and Crocodylus moreletii (Gonzalez-Trujillo et al.

2014) decreased in modified habitats unless a strip of

vegetation was present near the water bodies. In Nayarit,

Mexico, Hernandez-Hurtado et al. (2011) reported high

densities of all sizes of C. acutus, which were associated

with sites of low salinity, depths exceeding 1 m, and six

types of vegetation, conditions in which the crocodiles

presented greater dispersion and benefited from a higher

availability of food and shelter.

The coastal area of Colima, Mexico, has undergone

changes due to extensive construction of infrastructure.

Over a period of 13 years, 145.40 ha of mangrove

have been lost, out of a total area of 494.02 ha, which

Amphib. Reptile Conserv.

has modified the physical and biological structure of

the coastal ecosystems (Jiménez-Ramon et al. 2016).

While it 1s likely that a consequent modification of the

crocodile habitat could have occurred, the biotic and

abiotic variables related to the density and distribution of

American Crocodile populations that inhabit the area are

unknown. Therefore, the objective of the present study

was to evaluate the state of the crocodile populations in

Colima, Mexico, and to determine which habitat variables

influence the abundance, structure, and distribution of

this species. A greater crocodile abundance was expected

at lower salinities, as well as a greater density of adult

crocodiles in areas with higher temperatures, and higher

numbers of hatchlings and juveniles in areas with a

greater presence of aquatic vegetation.

Materials and Methods

Study area. The coastal zone of the state of Colima,

which comprises the study area, presents annual

temperatures ranging between 20 and 28 °C and mean

annual precipitation of 947.8 mm. The rainy season

begins in June, and precipitation events can be torrential,

especially at the end of August and the beginning

of September, while the rains begin to decrease in

November (Arévalo et al. 2016). This region supports

vegetation communities of low elevation dry deciduous

forest, scrub forest, and mangrove forest. Lagoons

with a known presence of crocodiles were selected for

this study, based on information provided by the local

environmental authorities and communities established

close to each lagoon. Six lagoons were selected, with the

largest (Laguna de Cuyutlan) divided into four sections

that communicated directly with each other (see Fig. 1

and Table 1 for lagoon abbreviations). The dominant

vegetation found in the selected lagoons is mangrove

forest comprising the species Rhizophora mangle and

Laguncularia racemosa, with secondary vegetation such

as Acacia sp., and floating vegetation such as Zypha

domingensis and Echhornia crassipes. It should be

noted that some lagoons are permanently connected to

the ocean (JU, C1, C2, C3), while others are connected

for only part of the year (VG, T, CH), and some are

completely disconnected from the ocean (C4 and A).

Sampling data. Night visits were made twice during the

rainy season (once in June 2014 and once in November

2015) and twice during the dry season (once in March

2015 and once in May 2016). The nocturnal spotlight

technique, as proposed by Messel et al. (1981) and

Thorbjarnarson et al. (2000), was used to determine the

relative population density, reported as individuals per

km (ind/km). From a 25 hp outboard motor boat, either

a 500-lumen headlamp or a handheld torch was used to

illuminate the water surface, the lagoon edges, below the

mangrove vegetation, and the emergent vegetation. The

individuals counted were grouped into size classes I to V

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Crocodylus acutus population structure in Colima, Mexico

Table 1. Identification key of study sites and environmental variable value averages for each site during the two seasons. Ta: water

temperature; Tai: ambient temperature; Sal: salinity; P: depth.

; : Ta Tai P

Key Lagoon/Estuary Latitude Longitude Sal (psu) (°C) (°C) (m)

JU Juluapan 19°06’50” 104°24’23” 24.9 28.7 25.9 1.9

VG Valle de las Garzas 19°05°17” 104°18°20” ps 28.9 27 1.3

all Cuyutlan Vaso I 19°02’ 13” 104°19718” 32.5 28.8 25.4 1.6

C2 Cuyutlan Vaso II ROSO 1212 104°15°30” 32.1 28.2 25.4 Lal

C3 Cuyutlan Vaso III 19°00’ 15” 104°12°53” 30.1 31.3 28.3 1.1

C4 Cuyutlan Vaso IV 18°54’05” TLO420 153°" 0.5 30.4 Del 0.8

T Tecuanillo 18°48°49” 103°53’52” 0.5 26.4 25.4 1.4

CH Chupadero 18°45’°01” 103°48’08” 0.6 29.3 24.8 1.4

A Amela 18950? 21" 103°45°49” 0.3 32 27.4 a5)

according to total body length, which was estimated by

calculating the distance from the base of the eyes to the

end of the snout of the crocodile and multiplying by ten

(Platt and Thorbjarnarson 2000).

Temperature and salinity were recorded at every km

traveled onthe lagoons. These parameters were measured

with a multiparametric PCSTestr35 (OAKTON, Vernon

Hills, Hlinois, USA), with a salinity meter accuracy of

+1.0% FS and automatic temperature compensation.

Depth was measured with an aluminum leveling rod 10

m in length, graduated in centimeters. Vegetation type

was the only biotic factor recorded; each lagoon was

visited during the day in order to describe the species of

flora present, which were grouped into four vegetation

types based on the classification by Ramirez-Delgadillo

and Cupul-Magafia (1999). The distance covered by

each vegetation type along the shore of each lagoon

for each km traveled was measured with a GPS

brand Garmin model e-trex 10, in order to determine

a relationship between the vegetation data and the

presence of crocodiles.

Data analysis and statistics. To determine the degree

of association between crocodile density, abiotic factors

(water temperature, ambient temperature, depth, and

salinity), and vegetation, a non-metric multidimensional

scaling analysis (NMDS) was performed in the program

PAST version 3.08 (Hammer et al. 2001), using the

average value of each variable obtained in each selected

lagoon. A dissimilarity matrix was obtained using the

Bray-Curtis coefficient (Hammer et al. 2001). This

analysis creates a stress coefficient, for which a value

below 0.1 indicates that the groups formed differ in

their compositions. To corroborate the groups obtained,

a dendrogram was created using the Bray-Curtis

coefficient (Hammer et al. 2001). This dendrogram

provided a correlation coefficient which indicated the

similarity among the lagoons, with values close to 1

indicating greater similarity.

Differences in crocodile density between seasons,

lagoons, and size classes were estimated with a

Kruskal-Wallis test (P < 0.05). This same analysis

was used to evaluate differences in crocodile densities

between the lagoons that were connected to each other,

and between lagoons that were either always connected

Amphib. Reptile Conserv.

to the ocean, connected for only part of the year, or

completely disconnected from the ocean. Differences

between abiotic factors per season and per lagoon

were evaluated with a Kruskal-Wallis test (P < 0.05),

and a Tukey test (P < 0.05) was used to identify where

differences occurred. This same analysis was used to

evaluate differences in vegetation per type and per

lagoon. All of these analyses were performed using the

software package PAST version 3.08 (Hammer et al.

2001).

Results

The study was conducted in a total of six lagoons that

collectively represented a surface area of approximately

9,545 ha, and were located along 76.6 linear km of the

coastline of Colima state. Significant differences were

found in the crocodile densities in these lagoons in both

seasons (P = 0.0001); the Tukey test indicated that the

differences occurred between the lagoons with greater

crocodile density (A and VG) and those with lower

density (C1, C2, and T). The NMDS analysis and the

dendrogram integrated the data from both seasons and

generated a stress coefficient of 0.06 and a correlation

coefficient of 0.92. There was a separation between the

lagoons connected to the ocean (JU, Cl, C2, and C3)

as one group, and those that were either not connected

to the ocean or only connected for part of the year

(VG, CH, and T) as another group (Fig. 2). However,

the dendrogram showed that lagoon C4 was similar to

the group of lagoons not connected to the ocean, while

lagoon A was dissimilar to all the others (Fig. 3).

Crocodiles were observed at the nine study sites

during both seasons. Relative frequencies ranged from

0.2—8.3 ind/km and 0—5.9 ind/km during the rainy and

dry seasons, respectively (Fig. 4). There were significant

differences in density in the A and CH lagoons during the

two periods (P = 0.0001), but no differences were found

between size classes (P = 0.4).

The density of individuals in the lagoons connected to

the ocean (JU, C1, C2, C3) was lower than in those not

connected to the ocean or connected for only part of the

year (P = 0.006). Densities of 0 to 1 ind/km in size classes

I, II, and II, and of O to 0.4 ind/km in size classes IV and V,

were recorded in the lagoons connected to the ocean for

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Aguilar-Olguin et al.

0.20

0.16

0.12

0.08

0.04

0.00

-0.04

-0.08

-0.12

-0.16

-0.20

-0.40 -0.30 -0.20 -0.10 0.00 0.10 0.20 0.30 0.40 0.50 0.60

Coordinate 1

Coordinate 2

Fig. 2. Non-metric Multidimensional Scaling (NMDS) analysis

showing the formation of two groups, by taking into account

the crocodiles observed, water salinity, temperature, depth,

and the four vegetation types present. Acronym definitions and

characteristics of the sites are given in Table 1.

only part of the year (Table 2). Eight species of flora were

identified on the shores of the sampling sites, all of which

were included in the vegetation classification proposed

by Ramirez-Delgadillo and Cupul-Magafia (1999).

Rhizophora mangle and L. racemosa were classified

as mangrove forest species, 7’ dominguensis and E.

crassipes as aquatic vegetation, Acacia sp. and Guazuma

ulmifolia as secondary vegetation, and Batis maritima

and Cocos nucifera as pasture-agricultural vegetation.

Of the 78.5 linear km traveled, mangrove vegetation

was present in eight of the nine lagoons, representing

an overall presence of 72%. Site A did not present

mangrove vegetation; however, 61% of the banks of this

lagoon presented aquatic vegetation and 39% presented

secondary vegetation. Of the total crocodile population,

55.4% was recorded in the lagoons where mangrove

vegetation occurred, while the remaining crocodiles

were recorded in aquatic vegetation (Table 3). There

were significant differences in crocodile density between

vegetation types (P = 0.003). The Tukey test showed that

the mangrove vegetation differed significantly (P < 0.05)

in this regard from the other vegetation types.

Salinity ranged from 0.2—27% during the rainy season,

and from 0.4—35.3% during the dry season. There were

significant differences in this parameter between the two

seasons (P = 0.001) and between the averages of lagoons

(P = 0.0002); and differences were also detected (P <

0.05) between the lagoons connected to the ocean (JU,

Distance

[e)

Fig. 3. Dendrogram considering the crocodiles observed, water

salinity, temperature, depth, and the four vegetation types

present. Acronym definitions and characteristics of the sites are

given in Table 1.

Cl, C2, C3) and those that were not connected to the

ocean (VG, C4, T, CH, A). Water temperature ranged

from 25.8—32.1 °C during the rainy season, and from

27.9-31.8 °C during the dry season, with a significant

difference found between the two seasons (P = 0.0002).

The ambient temperature ranged from 22.1—29.3 °C

during the rainy season, and from 26.3—28.2 °C during the

dry season, also with a significant difference between the

two seasons (P = 0.0002). The depth range was 0.9-3.5

m during the rainy season and 0.6—3.6 m during the dry

season; but there was no significant difference between

the two seasons (P = 0.7) in this regard, although there

were differences (P = 0.0005) in depth between lagoons.

Lagoon A differed significantly (P < 0.05) from the other

lagoons (JU, VG, C1, C2, C3, C4, T, CH).

Discussion

Density

The crocodile density ranges found in this study (0-8.3

ind/km) during the two seasons in the nine lagoons

can be considered low. This is due to the influences of

salinity, ambient temperature, water temperature, and

Table 2. Crocodile population structure expressed as relative density (individuals/km) and percentages (%) for each lagoon, in the

two study seasons. Only eyes indicates individuals for which the size could not be determined.

Size classes

Lagoon I II Il IV Vv Only eyes

I/km % I/km % I/km % I/km % I/km % I/km %

JU 0 0 0.9 22.4 0.7 15.3 [eal 34.1 0.3 10.6 0.6 17.6

VG 0.5 12 1.8 26.4 Ley 24.1 0.8 12 0.4 6 7 24.1

Cl 0 0 0.1 1 0 0 0 0 0 0 0 0

C2 0.3 35-5 0.2 29.4 0.3 35.3 0 0 0 0 0 0

C3 0.2 7 1 32.8 Ory oA Vie’ 0.6 20.4 0.4 11,3 0.2 -

C4 0.6 pe 0.6 29.1 0.5 OO. 0.3 12.1 0.1 4.3 0.1 5

T 0 0 0 0 0 0 1.8 1 0 0 0 0

CH 1 30.3 0.3 7.9 1 31.6 0.4 13.2 0.4 11.8 0.2 5.3

A 0.9 13 0.4 5.7 1 rs 1.8 26.8 1.5 22 1 17.5

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Crocodylus acutus population structure in Colima, Mexico

Rainy

Bm Dry

Relative density (individuals/km)

OrRPFN wo Psftu DN CO WO

A VG C4 C3 JU CH T C2 C1

Lagoon

Fig. 4. Average relative Crocodylus acutus density in the

studied lagoons during the two (rainy and dry) study seasons.

Acronym definitions and characteristics of the sites are given

in Table 1.

the mangrove vegetation, which varied depending on the

connection with the sea in lagoons JU, C1, C2, and C3.

This density level can be compared to four other similar

studies in different parts of Mexico.

In Chamela-Cuixmala, Jalisco, Garcia et al. (2010)

recorded 20.4—32.5 ind/km, classifying this density as

high and attributing it to variables such as water depth

(0.5 m) and mangrove vegetation cover.

In Boca Negra, Jalisco, Cupul-Magafia et al. (2002)

reported 51.2 ind/km and classified this density as

high, attributing it to the fact that the area provided

the crocodile population with refuge and feeding areas

through the presence of mangrove vegetation.

In Palma Sola, Oaxaca, Garcia-Grajales and Buen-

rostro-Silva (2014) reported a density of 70 ind/km,

classifying this value as high and attributing it to the

presence of the mangrove vegetation which provided an

important refuge area for the crocodiles.

In San Blas, Nayarit, Hernandez-Hurtado et al. (2011)

reported densities of 0.36—4.31 ind/km, which were

within the ranges found in this study, with the highest

densities found in sites with depths greater than 1 m, slow

currents, low salinities (4.92—11.03%), high elevations,

and six vegetation types, including mangrove swamp.

Thorbjarnarson et al. (2006) designated an area

of 22,790 ha as a bioregion of the coast of Colima

and Jalisco. They considered the vegetation type as

an important variable and estimated a population of

between 500 and 1,000 crocodiles. That assessment does

not coincide with the results of this study, since in less

than half of that area (9,945 ha belonging to the coast of

Colima), 506 crocodiles could be observed. However, the

difference may be due to the fact that the previous study

did not include any of the specific lagoons examined in

the present study.

Population Age Structure

The age structure observed in the present study (Table

2) suggests that the populations of crocodiles in Colima

are in good health, since the highest percentage of the

population belonged to classes I, II, and III, which are

classified as juveniles and subadults (Seijas 2011).

According to Calverley and Downs (2014), a population

structure where the largest number of crocodiles is

represented by juveniles and subadults can indicate a

positive trend, whereas a population where the largest

number of crocodiles is adults indicates a population

in decline. In this study, fewer size class HI crocodiles

were found in lagoons JU, VG, C4, and C3 (Table 2),

probably due to dispersion behavior (Cedefio and Pérez

2010; Hernandez-Hurtado et al. 2011; Garcia-Grajales

and Buenrostro-Silva 2014). This suggests that size class

IV and V crocodiles dispersed towards the vegetation

because of their territorial behavior, and remained there

in refuge without moving towards the open areas of the

lagoon (Carvajal et al. 2005; Balaguera and Gonzalez

2008).

Vegetation Types

In this study, mangrove was the predominant vegetation

and it was where the highest percentage of crocodiles

was recorded. This suggests that C. acutus finds refuge

and food in the mangrove vegetation in Veracruz.

Gonzalez-Trujillo et al. (2014) reported the density

Table 3. Linear extents (km) and percentages of vegetation types found in each lagoon.

Lagoon Mangrove Aquatic vegetation

km (“%) km (%)

JU 6 (100) 0

VG 1.5 (50) 0

Cl 6 (100) 0

C2 15.5 (97) 0

C3 16 (100) 0

ct 4 (67) 2 (33)

T 0.564 (100) 0

CH 7 (100) 0

is 0 11 (61)

Amphib. Reptile Conserv.

Secondary vegetation Pasture-agriculture

km (%) km (%)

0) 0)

0) 1.5 (50)

0) 0

0 0.3 (3)

0) 0)

0 0

0 0)

7 (39) 0)

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Aguilar-Olguin et al.

of Crocodylus moreletii was related to vegetation

type, with mangrove hosting the largest number of

crocodiles and attributed this to the vertical structure of

the mangrove, which provides suitable shelter. For C.

acutus in Oaxaca, Garcia-Grajales and Buenrostro-Silva

(2014) recorded the largest number of crocodiles in the

mangrove vegetation and attributed this to the fact that

crocodiles prefer to remain hidden among the roots of

the mangroves. In Ecuador, Carvayal et al. (2005) stated

that, in addition to providing refuge for the crocodiles,

the mangrove vegetation also provides food resources in

the form of invertebrates and vertebrates that also inhabit

the mangrove.

However, in Florida, Cherkiss et al. (2011) report

that mangrove is the habitat used least by C. acutus

crocodiles, since the largest number of individuals was

observed in artificial ponds, canals, and streams, which

offer more protection and this species can easily adapt to

anthropogenically created or transformed habitats. In the

present study, the Amela (A) lagoon is the largest and has

the most abundant aquatic vegetation where the highest

densities of crocodiles were recorded.

Abiotic Factors: Salinity, Connection to Ocean,

Depth, and Temperature

Salinity was an important factor in crocodile abundance

in the different water bodies. The lagoons with the highest

salinity formed one group and those with lowest salinity

formed another group (Table 1; Figs. 2 and 4). Salinity

could negatively affect the crocodile populations,

particularly the young (classes I and II) and juveniles

(class III), because high salinity causes the individuals of

this species to dehydrate, unlike C. porosus in Australia

which has two glands in the palate that allow it to expel

excess salt (Richards et al. 2004). The salinity-sensitivity

of this species was reflected in crocodile densities, since

the lowest values were recorded in the lagoons with high

salinity (Fig. 3). Hernandez-Hurtado et al. (2011) also

reported a decrease in crocodile density when salinity

increased (24.76—-35.85%). An increase in crocodile

density was observed here from the northern (C1) to the

southern (C4) sites sampled within the Cuyutlan Lagoon

(C1, C2, C3, and C4). At the latter site, salinity had

decreased to concentrations of 0.4-0.7% and juvenile

and adult crocodiles (classes I, II, and HI) were dominant

(Table 2). This could be due to the fact that hatchlings

and juveniles are the most vulnerable to high salinity

concentrations, which would prompt them to seek out

sites of lower salinity (Richards et al. 2004).

Dispersion of crocodiles towards different areas in

search of freshwater has been reported (Dunson and

Mazzotti 1989). A preference of crocodiles for areas with

salinity below 1% has also been observed (Espinosa et

al. 2012). The effect of salinity on crocodiles in classes I

and II could be fatal, since hatchlings and juveniles have

little tolerance for high salinity (Lang 1987). While some

Amphib. Reptile Conserv.

of the highest salinities in this study were observed at

lagoon C2, a large number of hatchlings were recorded

there (Fig. 3). However, these animals would die rapidly

due to dehydration if they did not move to sites with lower

salinity; and this was reflected in the relative densities of

classes IV (0 ind/km) and V (0.5 ind/km) [Richards et al.

2004; Hernandez-Hurtado 2010].

While lagoon JU was permanently connected to the

ocean, crocodiles of all classes, other than class I, were

observed there, and they were also observed very close

to the mouth of the Miramar Creek. The crocodiles in

size classes I, I, and III can avoid the high salinity of

the lagoon by finding refuge up this freshwater creek

during the rainy season, where they can reach adult sizes.

Individuals of C. acutus have been reported up this creek

(at elevations up to 1,220 m asl) between Guerrero and

Oaxaca (Casas-Andreu et al. 1990).

Temperature variations ranged from 3-6 °C (Table 1) in

the two study seasons. Carvajal et al. (2005) reported that

high temperatures (40 °C) influenced the distribution of

crocodiles, which preferred lower temperatures; whereas

Espinosa et al. (2012) reported that low temperatures

(14.42 °C) influenced the distribution of crocodiles,

which were observed in areas with higher temperatures.

Lang (1987) reported a positive correlation between

water temperature and crocodile density, as water and

air temperatures influence the body temperature of

crocodiles with implications for their physiological

processes. The relatively minor temperature variations

observed during this study probably did not affect the

normal crocodile thermoregulation processes. This could

be due to the fact that crocodiles living in environments

with little temperature variability adopt a strategy of

thermoconformity. In order to control this physiological

process, the crocodiles spend a large part of the day and

night in the water regulating their body temperature, a

process in which water depth can play an important role

(Campos and Magnusson 2012; Grigg et al. 1998; Lang

1987).

The depths observed in the present study ranged from

0.6—-1.92 m (Fig. 3), with significant differences in depth

among the lagoons. Lagoon A differed significantly from

the others, presenting both the greatest depths and highest

crocodile density. However, the depths recorded in all of

the lagoons were sufficient for the crocodiles to move,

reproduce, and feed (Fujisaki et al. 2009). Calverley and

Downs (2014) stated that variations in depth could affect

the number and distribution of crocodiles. Hernandez-

Hurtado et al. (2011) recorded the greatest number of

crocodiles at depths of over 1 m, which corroborated the

finding by Campbell et al. (2010) of a high correlation

between water depth and the distance traveled by

crocodiles towards feeding areas. Fujisaki et al. (2009)

reported that depth had a direct influence on adults during

the reproductive period, since both males and females

seek water depths of over 1 m in order to mate.

Lagoon A had no similarities with any of the

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Crocodylus acutus population structure in Colima, Mexico

other lagoons during both seasons. This lagoon was

characterized by having mostly aquatic vegetation and

abiotic factors that remained rather constant, with water

temperature variations of 1 °C, air temperature variations

of 2 °C, as well as the greatest depths, and freshwater

available throughout the year. The distribution and state

of health of the crocodiles at this location were dictated

by the low variation in the abiotic factors and aquatic

vegetation that provided refuge and food. Moreover,

in this case, intraspecific relationships had a greater

influence on the population (Lang 1987; Grigg et al.

1998; Balaguera and Gonzalez 2008; Cherkiss et al.

2011).

Conclusions

In the present study, the abiotic and biotic factors

evaluated, including salinity, ambient temperature, water

temperature, and mangrove vegetation, were found to

have close relationships with the population dynamics

of the crocodiles in the State of Colima, and changes in

these variables may influence the distribution of C. acutus

populations. High salinities may restrict the crocodiles to

sites that are supplied with fresh water, either constantly

or for some part of the year. These high salinities can

have direct effects on organisms, particularly in the early

stages of their development. Continued monitoring of

these variables, as well as the populations of crocodiles,

should contribute to our understanding of the behavior of

these populations of crocodiles prior to modification of

their habitat, as well as determining the characteristics of

the habitat they prefer. These are priorities for contributing

to the improved management and conservation of this

species.

Acknowledgments.—We thank the Unidad de Manejo

de Fauna Silvestre CEC, Universidad de Guadalajara,

and CONACyT for the postgraduate scholarship

450956/292358 granted to SAO. Thanks also go to the

team that helped during fieldwork: Jonathan Ceya, Judit

Torres, Mayra A. Machuca, Jaime Thomas, and Francisco

Ifiguez.

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Darwin, Northern — Territory,

Sergio Aguilar-Olguin has a degree in Oceanography and a Master’s degree in Marine Sciences, both

from the School of Marine Sciences, Universidad de Colima (Mexico). Sergio is currently pursuing

a Doctorate in Ecology and Management of Natural Resources at the University Center of the South

Coast, Universidad de Guadalajara (Mexico), and is a professor for the honors degree in Biology in the

Faculty of Biological and Agricultural Sciences, Universidad de Colima. Since 2001, Sergio has worked

as head of the Department of Marine Turtles for the Management Unit, Conservation of Wildlife (UMA)

Ecological Center of Cuyutlan “El Tortugario,” where he coordinates activities on the protection and

conservation of sea turtles and conducts studies on the population and reproductive ecology of sea

turtles and crocodiles.

Maria Cruz Rivera Rodriguez is a graduate of the Faculty of Biological Sciences, Michoacan

University of San Nicolas de Hidalgo, Mexico, with a Doctorate in Animal Science from the

University of Colima, Mexico, with specializations in Ecology and Environmental Impact, as well

as Environmental Education. Maria has served as Technical Manager of the UMA Ecological Center

of Cuyutlan “El Tortugario,” and a professor-researcher of the Faculty of Biological and Agricultural

Sciences, University of Colima. She is a coordinator of research projects on wildlife management and

conservation related to crocodiles, iguanas, sea turtles, snakes, lizards, and birds.

Helios Hernandez-Hurtado is a Biologist who graduated from Universidad de

Guadalajara, and received his Ph.D. from Universidad Autonoma de Nayarit, México.

Helios has provided technical support to Reptilario Cipactli at Centro Universitario de la

Costa, Universidad de Guadalajara (México) and is an active member of the Crocodile

Specialist Group of IUCN. For 22 years, Helios has been working on the research,

management, and conservation of crocodiles in captivity, as well as related projects

on population ecology and management in the wilderness, in collaboration with local

communities.

Ricardo Gonzalez-Trujillo is a Biologist with a Ph.D. in Science in the area of Ecology and Natural

Resource Management. Currently, Ricardo applies population genetics tools to solve problems related

to ecology and the use and management of natural resources.

Maria Magdalena Ramirez-Martinez is currently a professor and researcher at Universidad de

Guadalajara in Jalisco, México. Her research interests concern the ecology and biology of vertebrates,

focusing on population dynamics and conservation, as well as the ecology of zoonotic diseases in

western México. Maria enjoys traveling, long walks, and dance.

182 April 2020 | Volume 14 | Number 1 | e231

Official journal website:

amphibian-reptile-conservation.org

Amphibian & Reptile Conservation

14(1) [General Section]: 183-189 (e232).

More road-killed Caspian Whipsnakes (Dolichophis casptius):

an update on the species distribution along the Danube,

in Romania

*Severus-D. Covaciu-Marcov, Alfred-S. Cicort-Lucaciu, Daniel-R. Pop,

Bogdan I. Lucaci, and Sara Ferenti

University of Oradea, Faculty of Informatics and Sciences, Department of Biology, Universitatii str. 1, 410087 Oradea, ROMANIA

Abstract.—Dolichophis caspius is present in Romania only in the southern regions, which partially represent

its northern distribution limit. In recent years, new data on the distribution of D. caspius in the country have

been collected, with many of these records being road-killed individuals. Conducted during 2013-2019, this

survey further completes the information on this species’ distribution in southern Romania, recording 55

(mostly new) distribution localities. Dolichophis caspius is a constant presence in the Danube meadow, and it

also advances northwards along the Jiu, Olt, and Arges River meadows. Dolichophis caspius was mostly found

to inhabit areas with loess walls, but it was also present in flat areas with agricultural terrains. Most of these

records were road-killed individuals. The high number of new distribution records could be driven by the last

year’s weather conditions, which were unusually warm and dry in the region. Also, the high number of road-

killed snakes clearly reflects the increasing level of the human pressure on D. caspius in a region where natural

habitats are becoming increasingly rare.

Keywords. Climate, Colubridae, habitat, loess walls, Reptilia, road mortality, Serpentes

Citation: Covaciu-Marcov SD, Cicort-Lucaciu AS, Pop DR, Lucaci BL, Ferenti S. 2020. More road-killed Caspian Whipsnakes (Dolichophis caspius):

an update on the species distribution along the Danube, in Romania. Amphibian & Reptile Conservation 14(1) [General Section]: 183-189 (e232).

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: 4 February 2019; Accepted: 20 February 2020; Published: 20 April 2020

Introduction

The Caspian Whipsnake (Dolichophis caspius) is an

Eastern-Mediterranean species (e.g., Tomovic et al.

2014), present in Europe in its south-eastern regions, the

Italian Peninsula, and the Balkan Peninsula (Sillero et al.

2014). A large part of its distribution range was probably

populated shortly after the last glacial maximum (Nagy et

al. 2010). The north-western limit of its distribution range

reaches Hungary and Romania (see Nagy et al. 2010;

Sahlean et al. 2014). In Hungary, it occupies only a very

small territory near the Danube River, where it ascends

farther north than in Romania, and in the last years new

distribution records have been reported (e.g., Korsos et

al. 2002; Bellaagh et al. 2008). Nevertheless, even the

largest population in Hungary seems to be in decline

(Frank et al. 2012). In Romania, D. caspius is present

only in the southern part of the country, along the Danube

River, especially in Dobruja (see Cogalniceanu et al.

2013; Sahlean et al. 2019). Its populations are considered

to be declining in the country, with some regarded

Correspondence. *severcovaciul@gmail.com

Amphib. Reptile Conserv.

as extinct (Iftime 2001-2002). The new distribution

records of the last decades (Strugariu and Gherghel

2007; Covaciu-Marcov and David 2010; Sahlean et al.

2010; Ferenti et al. 2011; Covaciu-Marcov et al. 2012a)

updated the knowledge on the species distribution in

Romania (Cogalniceanu et al. 2013). However, D.

caspius was still recorded in new localities in more recent

years (Iftime and Iftime 2015, 2017; Sahlean et al. 2019),

while large gaps in its known distribution range still

remain (Sahlean et al. 2019). The majority of the recent

distribution records represent road-killed individuals

(e.g., Covaciu-Marcov and David 2010; Ferenti et al.

2011; Covaciu-Marcov et al. 2012a). Although these

data indicate that D. caspius has a broader distribution

range in Romania than was previously considered (Fuhn

and Vancea 1961), they also confirm the pressure under

which it is being subjected (Iftime 2001-2002). In the

light of these facts, records of road-killed whipsnakes are

of particular interest. Therefore, this note is an update on

D. caspius distribution and road mortality intensity along

the Danube in southern Romania.

April 2020 | Volume 14 | Number 1 | e232

Caspian Whipsnake distribution in Romania

Bucharest

Fig. 1. Dolichophis caspius distribution in southern Romania.

Squares: previous records (after Cogalniceanu et al. 2013;

Iftime and Iftime 2016, 2017), red dots: new records.

Materials and Methods

The information on the distribution of D. caspius in

southern Romania was obtained from field studies realized

between 2013 and 2019. In 2016, 2017, and 2018, the

Danube meadow between Drobeta-Turnu Severin and

Fetesti was surveyed once each year, in spring or in autumn.

Additional surveys occurred 2—3 times each year in the

western sector, from Drobeta-Turnu Severin to Bechet,

and 1—2 times each year in the Jiu River meadow, between

Craiova and Bechet. In total, ~29 days were spent in areas

which could shelter D. caspius populations. None of those

days were dedicated specifically to D. caspius, and snakes

were observed by chance during other field work in the

region (e.g., Covaciu-Marcov et al. 2017a,b, 2018). Road-

killed individuals were observed directly from the car by

driving at speeds of 50-60 km/h on roads with low traffic,

a method previously used in the region (Covaciu-Marcov

et al. 2012a). In some cases, direct searches for snakes

were conducted in favorable habitats, and the observed

individuals were recorded and photographed. Road-

killed individuals were also photographed, and those in

better physical condition were collected and deposited

in the zoological collection of the University of Oradea,

Romania.

Results

Dolichophis caspius individuals were recorded from 55

distribution localities in southern Romania (Table 1), most

of which (52) were new distribution records (Fig. 1). The

majority of the new distribution records are located in the

western part of southern Romania, in the historical region

of Oltenia, in Mehedinti and Dolj counties. Overall, 73

individual D. caspius snakes were identified. Five of the

whipsnakes were encountered alive, while the others

were found killed either by humans (two individuals), or

by cars on the roads (Fig. 2). Juveniles and adults of up

Amphib. Reptile Conserv.

R bi MS Meh, eee iret That

Fig. 2. Road killed Dolichophis caspius individual from

Listeva, Dolj County, Romania.

to 1.60 m total length were both found killed on roads.

Dolichophis caspius individuals were mostly found in

spring (April-May) and autumn (September—October).

Discussion

Compared to the previously available data (e.g.,

Cogalniceanu et al. 2013; Iftime and Iftime 2015, 2017;

Sahlean et al. 2019), the results reported here more than

double the number of D. caspius records in the Danube

meadow. This confirms once again that the species

presents a continuous distribution in the region (e.g.,

Ferenti et al. 2011; Covaciu-Marcov et al. 2012a). While

this fact was already well-established in the central

part of the meadow (Sahlean et al. 2010; Ferenti et al.

2011; Cogalniceanu et al. 2013; Iftime and Iftime 2015,

2017), in the western parts the new data completed the

D. caspius distribution range between Drobeta-Turnu

Severin and the Olt River (Fig. 1), where a large gap

has existed until now (Sahlean et al. 2019). These new

records also show a continuous distribution along the

Jiu River between Bechet and Craiova, and confirm the

sporadic presence of this species north of Craiova (e.g.,

Sahlean et al. 2019). Although the three previously known

distribution localities along the Jiu River already indicated

this, Caspian Whipsnakes were rarely recorded in the

region (e.g., Sahlean et al. 2019), despite the presence of

apparently favorable habitats. The whipsnakes are also

continuously distributed between Olt and Jiu Rivers. Based

on only four records, D. caspius seems less common in

the area between Calafat and Bechet. The records from the

Arges River region confirm that the species is currently

well represented, although it was previously considered

extinct from one locality (Iftime 2001—2002).

April 2020 | Volume 14 | Number 1 | e232

Covaciu-Marcov et al.

Table 1. Dolichophis caspius distribution records in southern Romania. Asterisks (*) indicate previous records. County abbreviations:

Mh = Mehedinti County, Gr = Giurgiu County, Cl = Calarasi County, Tr = Teleorman County, Ot = Olt County, Dj = Dolj County,

Il = Ialomita County.

‘ p : Road- Human ‘

Locality Date(s) Geographic coordinates killed killed

aera oe 2 VI 2018 1

1 Mh : 0 ee : ae 44°39’28.90”N, 22°40°01.68"E

every ane 24 V 2015 2

3 VI 2018 E 4 y 3

Den Sees Coes

Pristol / Garla Mare} 21 VI 2016 | 44°13°08.71”N, 22°44°15.95"E] 1 [| - fe

Vanju Mare /Bucura | 10 V 2014 | 44°24’39.12”N, 22°51°52.83"E} 1 | - | - |

Arginesti / Butoiesti_| 28 VI 2015 | 44°34°39.33”N, 23°24°07.66"E} 1 | - | - |

Gruia / Patulele V2013 | 44°18°12.86”N, 22°44°27.07"E]| 1 | - | -

Balta Verde 30 1X 2019 | 44°20°17.70"N, 22°36°03.54"E] 1 | - | - |

Gr/Cl__| Greaca/Cascioarele* | 121V 2014 | 44°07°37.37°N, 26°25°04.89"E] - [| - | 1

13 X 2018, 0 > 0 >»

; nh 13 X 2018, ys 5 mies 09

13 X 2018 . > ° »

oa S124 [esoonrens ee

Amphib. Reptile Conserv. 185 April 2020 | Volume 14 | Number 1 | e232

Caspian Whipsnake distribution in Romania

Table 1 (continued). Dolichophis caspius distribution records in southern Romania. Asterisks (*) indicate previous records. County

abbreviations: Mh = Mehedinti County, Gr = Giurgiu County, Cl = Calarasi County, Tr = Teleorman County, Ot = Olt County, Dj

= Dolj County, I] = Ialomita County.

Road- Human

No. | County Locality Date(s) Geographic coordinates killed killed

ee a

Tas | D) | Bechet w the harbour | 14x 2017 [asrac'on 70". ss7a09E| 1 | - -

Taf pf sao | 2va0 [serie nasemoref 1 [|

ras [pi | titra | axa0is [esos ae nassssiaref 1 | - -

= a

30 VII 43°50’27. SPN 23°50 5737 .E

Zaval 019

53 Il Tandarei 28 VII 44°38°44.96 N, 27°41°06.00”E 1

; 2018

Tandarei / Mihail 25 VII 44°39’11.49”N, 27°42’29.19”E

54 Il ve 1

Kogalniceanu 2019

The new records confirm the association of D.

caspius with the loess walls of the Danube meadow (e.g.,

Ferenti et al. 2011; Covaciu-Marcov et al. 2012a). Most

observations were made in areas with loess walls, both

along the Danube meadow and its main tributaries (Jiu,

Olt, Arges). Along the Danube, loess walls have southern

exposure; while along the Jiu (Fig. 3) and Olt Rivers they

have western exposure, with southern oriented meadows,

and along the Arges River they are exposed to the north.

Only some localities, such as Macesu de Jos, are devoid

of loess walls, with a plain relief and an indistinguishable

boundary between the meadow and its neighboring

terraces. In such cases, agricultural lands surrounding the

roads were used by D. caspius, as is found in other parts

of its distribution area (e.g., Ioannidis and Bousbouras

1997; Arslan et al. 2018; Sahin and Afsar 2018). Thus,

the loess walls are confirmed to be favorable habitats for

D. caspius (e.g., Korsos et al. 2002; Bellaagh et al. 2008),

but some individuals persist even in flat habitats.

The data presented here extend the northern limit of the

distribution of D. caspius along the Danube tributaries.

Whipsnakes are xerophilous, influenced by temperature

and precipitation, and so the future northward distribution

could be facilitated by climate change (see Sahlean et

al. 2014), and even hatching time depends regionally

on temperature (Sahlean and Strugariu 2018). In south-

western Romania, the last few decades were warmer and

drier than longer term averages (Pravalie et al. 2014),

and D. caspius was mostly recorded in the regions of the

country with a drier climate (Matei et al. 2016). The high

Amphib. Reptile Conserv.

number of whipsnakes found could be a consequence of

favorable climatic conditions that enabled an increase in

the population growth rate. Moreover, D. caspius could

already be at an advantage due to climate change, despite

the high number of road-killed individuals. Nevertheless,

until recently the Danube meadow was mostly covered

by wet areas, lakes, or forests (probably except for the

loess walls), which are now drained, deforested, and

replaced with agricultural terrains (e.g., Licurici 2011;

Benecke et al. 2013; Geacu et al. 2018). Thus, agriculture

probably enlarged D. caspius habitats beyond the loess

walls. This scenario 1s supported by previous records of

this species in the agricultural areas of other regions as

well (e.g., loannidis and Bousbouras 1997; Arslan et al.

2018; Sahin and Afsar 2018).

The high number of D. caspius records is not

necessarily an indicator of favorable conditions, but

Fig. 3. Dolichophis caspius habitat inJ iu meadow, Romania.

April 2020 | Volume 14 | Number 1 | e232

Covaciu-Marcov et al.

clearly suggests a great human pressure in which

snakes are disturbed by intensive agriculture and habitat

alteration. In trying to avoid this impact, they move more

often, more frequently falling victim to the increasing

road traffic. Increased traffic intensity is known to

increase road mortality (e.g., Jones et al. 2014; Miranda

et al. 2017; Goncalves et al. 2018), even if D. caspius is

the fastest snake in Romania (Fuhn and Vancea 1961). In

addition, the greater activity of herpetologists may have

increased the number of records and observed road-killed

individuals. Nevertheless, we have no prior information

on the species status before the extensive alteration of

the Danube meadow (e.g., Licurici 2011; Benecke et al.

2013; Geacu et al. 2018). Dolichophis caspius records

based on road-killed individuals are not specific only to

Romania (Covaciu-Marcov and David 2010; Ferenti et al.

2011; Covaciu-Marcov et al. 2012a; Sahlean et al. 2019),

but are also commonly reported in other regions (e.g.,

Korsos et al. 2002; Krémar et al. 2007; Kambourova-

Ivanova et al. 2012; Mollov et al. 2013). Most of the

whipsnakes found were killed on roads situated at the

upper limit of the loess walls, just as previously indicated

(Covaciu-Marcov et al. 2012a). Road-killed individuals

were rarely recorded at the base of the loess walls, or in

regions without loess walls.

In southern Romania, D. caspius has arrived

only recently, at the end of its postglacial expansion

(Nagy et al. 2010). It uses natural and restrictive, but

also modified, habitats. Its protected status (O.U.G

57/2007, Monitorului Oficial al Romaniei, http://www.

monitoruloficial.ro/) increases the importance of the

regional biodiversity, where numerous relicts are present

(e.g., Pascovschi 1967; Ferenti and Covaciu-Marcov

2014; Covaciu-Marcov et al. 2017a, 2018). Dolichophis

caspius is not the only southern species with its northern

distribution limit in this region, but there are also other

amphibian and reptile species in the same category (e.g.,

Covaciu-Marcov et al. 2012b; Cogalniceanu et al. 2013;

Székely et al. 2013).

The records of D. caspius, especially road-killed

individuals, have been increasing in recent years in

Romania (e.g., Strugariu and Gherghel 2007; Covaciu-

Marcov and David 2010; Sahlean et al. 2010; Ferenti

et al. 2011; Covaciu-Marcov et al. 2012a; Iftime and

Iftime 2015, 2017; Sahlean et al. 2019). The finding of

numerous snakes is a consequence of what kills them,

namely the road network development. The impact of

roads on biodiversity is increasingly clear, including in

Romania (e.g., Cicort-Lucaciu et al. 2012, 2016; Ciolan

et al. 2017; Covaciu-Marcov et al. 2017c; Popovici

and Ile 2018; Teodor et al. 2019). The proposal of

speed reductions in regions populated by whipsnakes

(Covaciu-Marcov et al. 2012a) is no longer feasible

because, according to the new distribution records, the

speed limits would need to be applied to hundreds of km

of the roadway. If the protection of relicts strictly related

to natural habitats is relatively facile, depending on the

Amphib. Reptile Conserv.

conservation of the last remnants of initial habitats,

then the conservation of a species (such as D. caspius)

which recently arrived at its postglacial expansion (Nagy

et al. 2010) and is probably influenced by the climatic

changes (Sahlean et al. 2014) or even by some human

impacts, is much more difficult. This difficulty is further

compounded by the fact that this species could enter in

conflict with the exact factors that favor it at the moment.

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Severus-Daniel Covaciu-Marcov, Ph.D., is a Romanian biologist and author of faunistic studies on

herpetofauna, as well as other animal groups, from Romania. Many of his studies were realized in protected

areas. Sever is mainly interested in the biogeography of different animal groups, but also in the anthropic

impacts upon the fauna. In this regard, he has published studies on road mortality, invasive species, and

other issues. Sever is an Associate Professor at the University of Oradea, Faculty of Informatics and

Sciences, Department of Biology, in Oradea, Romania.

Alfred-Stefan Cicort-Lucaciu, Ph.D., is a lecturer at the University of Oradea, Faculty of Informatics

and Sciences, Department of Biology. His research is mainly focused on the biodiversity of wetlands,

while most of his publications concern herpetology. Alfred’s Ph.D. thesis focused on the biology, ecology,

and distribution of salamanders and newts in Romania.

Daniel-Razvan Pop is an associate assistant at the University of Oradea, Faculty of Informatics and

Sciences, Department of Biology, and a 1“ year Ph.D. student in the Doctoral School of Biomedical

Sciences, Domain Biology of the same university. Daniel’s studies are now focusing on railway ecology,

but he has also worked on studies concerning fauna distribution and anthropogenic impacts.

Bogdan-lIonut Lucaci is a 3 year B.Sc. student at the University of Oradea, Faculty of Informatics and

Sciences, Department of Biology. Bogdan is interested in wildlife ecology and biogeography, focusing

especially on certain groups of vertebrates, such as amphibians and reptiles. He is an enthusiastic

participant in different studies within the domains of biodiversity conservation in natural protected areas

Sara Ferenti, Ph.D., is a biologist from Romania, with a special interest in the geographic distribution

and ecology of terrestrial isopods and herpetofauna. She has conducted studies on the fauna of some

natural protected areas and some endemic species from Romania, but also on the anthropogenic impacts

upon the fauna. Sara is an Assistant Professor at the University of Oradea, Faculty of Informatics and

April 2020 | Volume 14 | Number 1 | e232

Amphibian & Reptile Conservation

14(1) [General Section]: 190-202 (e233).

Official journal website:

amphibian-reptile-conservation.org

Feminization tendency of Hawksbill Turtles (Eretmochelys

imbricata) in the western Yucatan Peninsula, Mexico

12*Cynthia Dinorah Flores-Aguirre, *Veronica Diaz-Hernandez, ‘Isaias Hazarmabeth Salgado Ugarte,

Luis Enrique Sosa Caballero, and 2*Fausto R. Méndez de la Cruz

'Posgrado en Ciencias Bioldgicas, Instituto de Biologia, Universidad Nacional Autonoma de México, A.P. 70-153. C.P. 04510, Ciudad de México,

MEXICO 24Instituto de Biologia, Departamento de Zoologia, Universidad Nacional Autonoma de México, Circuito exterior sin numero, Ciudad

Universitaria, Copilco, Coyoacdn, cp 04510, Ciudad de México, MEXICO 3Facultad de Medicina, Departamento de Embriologia, Universidad

Nacional Auténoma de México, Circuito Interior Ciudad Universitaria, Avenida Universidad n. 3000, cp 04510, Ciudad de México, MEXICO

“Facultad de Estudios Superiores Zaragoza, Campus dos, Batalla 5 de mayo s/n esquina Fuerte de Loreto, Colonia Ejército de Oriente, Iztapalapa

c.p. 09230, Ciudad de México, MEXICO

Abstract.—The viability of sea turtle populations during their early stages depends mostly on hatching success

and a balanced sex ratio of the hatchlings. In sea turtles, sex is determined by the temperature at which the

eggs are incubated. Consequently, climate change can play a critical role in population fitness, because

increased temperatures can skew the sex ratio towards females. The Hawksbill Turtle is a Critically Endangered

species and Mexico holds one of the largest populations of nesting females worldwide. Given the importance

of Mexican Hawksbill populations, it is necessary to evaluate their nest temperatures, particularly during the

thermosensitive period, and identify the sex ratio of the hatchlings. Hawksbill Turtle nests were characterized

in three nesting beaches from the western Yucatan Peninsula, Mexico, to determine the temperatures of the

nests in situ and the sex ratio of the hatchlings. The results showed that the incubation temperature was

warmer than the pivotal temperature by 1.255 + 0.18 (SE) °C, and in 82.05% of the nests monitored with only

female hatchlings. This was also confirmed via histology of the gonads of dead hatchlings. The amount of

shade above the nests and rainfall were the most influential factors for decreasing nest temperatures. This

study shows a trend toward the feminization of hatchlings due to the low percentage of shade above nests

which is able to increase the sex ratio bias due to coastal dune deforestation, together with the increase in

environmental temperatures due to climate change. The skewed ratio toward females could have negative

consequences for the maintenance of sea turtle populations on the Yucatan Peninsula, so is necessary to

develop strategies of conservation in order to reduce this trend.

Keywords. Anthropogenic impact, climate change, coastal-dune vegetation, female hatchlings, gonad histology, nest

temperature, pivotal temperature, rain, shade, temperature-dependent sex determination

Citation: Flores-Aguirre CD, Diaz-Hernandez V, Salgado Ugarte IH, Sosa Caballero LE, Méndez de la Cruz FR. 2020. Feminization tendency of

Hawksbill Turtles (Eretmochelys imbricata) in the western Yucatan Peninsula, Mexico. Amphibian & Reptile Conservation 14(1) [General Section]:

190-202 (e233).

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: 15 June 2019; Accepted: 17 February 2020; Published: 25 April 2020

Introduction pivotal temperature (PT), and increasing or decreasing

the temperature relative to the PT shifts the sexual

Temperature-dependent sex determination occurs in a

variety of reptiles, including all sea turtles (Bull and Vogt

1979; Wibbels 2003). In these species, the temperature

that embryos experience during their development

determines the sex of hatchlings. Specifically, sex

determination occurs in the thermosensitive period (TSP),

which 1s a particular time during embryonic development

(the middle third of incubation) when the temperature

affects gonad differentiation (Ciofi and Swingland 1997;

Mrosovsky and Pieau 1991). The TSP provides a balance

between both sexes at a certain temperature, called the

proportion towards one sex (Mrosovsky and Yntema

1980). In sea turtles, the sex ratio is skewed toward

males at temperatures below the PT and toward females

at temperatures above the PT (Salame-Méndez 1998).

Since the PT for all sea turtles is between 28 °C and 30

°C (Hawkes et al. 2009), the increase in temperature due

to global warming of the planet could skew the sex ratio.

A bias towards feminization has been reported in all

species of sea turtles in multiple studies around the world

since 1997 (Hawkes et al. 2009), being less represented

in Hawksbill Turtles (Eretmochelys imbricata, Hawkes

Correspondence. *“dcynthia,fa@gmail.com (CDFA), roveazdih@yahoo.com.mx (VDH); isalgado@unam.mx (IHSU); gigesc@gmail.com

(LESC); *faustor@ib.unam.mx (FRMC)

Amphib. Reptile Conserv.

April 2020 | Volume 14 | Number 1 | e233

Flores-Aguirre et al.

92°0'0O"W 97°0'O"W

Nest sampling month

@ June

A August

Yucatan

Campeche

2) Punta Xen

3) Chenkan

92°0'O"W 91°0'0"W

Kilometers 1:1,250,000

90°0'O"W

Fig. 1. Geographical locations of the monitored nests of Eretmochelys imbricata, as well as the beaches studied and their images.

1) Celestun, 2) Punta Xen, and 3) Chenkan.

et al. 2009; Hays et al. 2017); even though Hawksbills

are considered Critically Endangered (see https://www.

iucnredlist.org/search?taxonomies=1 008 88&searchType

=species). Thus far, no studies have addressed incubation

temperatures on Mexico’s nesting beaches, even though

the Yucatan Peninsula hosts one of the largest populations

of Hawksbill females worldwide and the largest nesting

population of Hawksbill Turtles in the Atlantic (Campbell

2014; Gardufio-Andrade and Guzman 1999; Meylan and

Donnelly 1999; Mortimer and Donnelly 2008).

Sea turtles select microenvironments for nesting,

thereby manipulating the thermal conditions to which

their eggs will be exposed during incubation (Diffenbaugh

and Field 2013). Thus, the nest temperature will be

modified according to its location with respect to several

factors, including vegetation (Kamel 2013; Kamel and

Mrosovsky 2006), distance to the coastline (Rees and

Margaritoulis 2004), sand differences in albedo and grain

size (Hays et al. 2001; Schaetzl and Anderson 2005), and

others. In addition, the thermal properties of the beaches

can provide different incubation temperatures (Hawkes

et al. 2009), and the temperature varies across the nesting

season (Mrosovsky et al. 1984).

Therefore, this study was carried out on three

beaches in the western Yucatan Peninsula with different

degrees of anthropogenic impact (disturbance due to

human infrastructures like roads, building for tourists,

vegetation removal, etc.) as well as variations in the

littoral characteristics (width of the beaches, distribution

and type of natural vegetation). These differences are

expected to promote diverse thermal gradients which

modify the nest temperatures, and, consequently, sex

ratios and hatching success. It is known that the least

Amphib. Reptile Conserv.

preserved beaches (especially in the dune strip) are

those that maintain the highest average temperatures

and decrease the hatching success (Pike 2008, 2009),

particularly for Hawksbill Turtles whose nesting is

closely related to vegetation (Horrocks and Scott 1991;

Kamel 2013). Therefore, the objectives of this study

were to determine the temperatures of Hawksbill Turtle

nests in situ during the TSP and estimate their sex ratios

based on a histological calibration, as well as determine

if there are differences in the sex ratio over the course of

the nesting season and among beaches.

Materials and Methods

Fieldwork was carried out on three index nesting beaches

(historical localities with higher nesting females) of the

Hawksbill Turtle in the western Yucatan Peninsula,

Mexico. The beaches were: Chenkan (19°07’50°N;

90°31°713.15°W) and Punta Xen (19°12’42.57°N;

90°52’07.01”W) in the state of Campeche, and Celestun

(20°59’33.72”N; 90°24’54”W) in the state of Yucatan

(Fig. 1). These beaches were selected as they present

differences in their littoral characteristics (width of the

beach and type of vegetation) and in their conservation

status (degree of disturbance due to human infrastructure).

Celestin is the most well-preserved beach (with wider

beaches, low disturbance in vegetation, limited erosion

and few human infrastructures on most of the beach),

while Chenkan is the least preserved (with high erosion

and deforestation rates, and a road along the beach).

The study was conducted in 2017, during the middle

(June-July) and end (August-September) of the nesting

season. June is the month when the nesting increases

April 2020 | Volume 14 | Number 1 | e233

Feminizaton of Eretmochelys imbricata in Mexico

(Guzman et al. 2008), so it would be more representative

to sample then, while in August the temperature begins

to decrease due to the increase in rainfall (Garcia 2004).

These differences could allow the evaluation of distinct

responses to the temperature during nesting, which would

modify the sex of the hatchlings and would therefore

provide greater variability in the samples.

The permit to handle the Hawksbill Turtles was

provided by SEMARNAT (SGPA/DGVS/03995/17).

The fieldwork consisted of making routes from 2100

to 0500 h, searching for nesting Hawksbill Turtles

randomly but trying to include different thermal zones

(i.e., under high vegetation (HV), under low vegetation

(LV), and without vegetation (WV), at different distances

along each beach). The study was conducted using only

nests in situ (1.e., places that females naturally selected to

lay their eggs), so once the turtles began to oviposit, the

nest location data were taken with a Global Positioning

System (GPS), as the nests were left exactly where the

turtles nested.

The characterization of each nest involved the

following procedure. The temperatures (°C), both

inside and outside the incubation chamber, were taken

with a digital thermometer (Fluke, 51-I[®). The depth

of the nest from the first to the last egg deposited was

measured with a flexometer. Temperature data loggers

(HOBO® pendant) were placed in the middle of the nests

and temperature data were recorded every half-hour

throughout the incubation period. The distances from

the nest to the nearest vegetation and to the coastline

were measured with a flexometer. The type of vegetation

closest to the nest was recorded, and the percentage of

shade above the nest was established when checking the

nest close to noon.

The hatching success was determined according to

Miller (1999), by checking the nests once the hatchlings

had left, after 55 + 5 d incubation (Guzman 2008). At the

hatchling stage, sea turtles cannot be sexed by external

morphology (Merchant 1999; Wibbels 2003). Therefore,

the sex ratios of the Hawksbill hatchlings were estimated

using the temperatures recorded by the data logger

during the TSP, which were analyzed by the TSD

software version 4.0.3 designed by Girondot (1999). In

addition, the temperature used to determine the sex in

the software was calibrated based on the histology of

the dead hatchlings, as this option avoided destructive

sampling of this Critically Endangered species. Any dead

hatchlings found in the monitored nests were collected

to dissect the urogenital complex (mesonephros and

gonads), which was fixed in 4% paraformaldehyde and

transported to the Laboratorio de Embriologia de la

Facultad de Medicina, Universidad Nacional Autonoma

de México. In the laboratory, the samples were dehydrated

by gradual ethanol solutions and maintained in 75%

ethanol. Later, they were embedded in _ histological

paraffin and the gonads were sectioned (between 8 and

12 um thick) using a rotary microtome. Subsequently,

Amphib. Reptile Conserv.

the samples were stained with periodic acid-Schiff’s

reaction and mounted on slides for observation under an

optical microscope. Under the microscope, the ovaries

or testes were identified using the criteria of Yntema and

Mrosovsky (1980) and Wibbels et al. (1999).

Sex ratio results obtained from gonad histology were

used as input for the TSD software to obtain a field

estimate of the PT for this species. Subsequently, this

PT was used as a formula parameter for estimating the

hatchling’s sex ratios, based on the mean incubation

temperature during TSP recorded in each nest. Since the

TSD software was developed for use with data derived

from constant temperature incubations, the average

temperature (and standard deviation) of each nest was

measured during each third of the incubation period,

in order to gauge the temperature variability within the

nests.

Since the shade provided by the HV decreases the

temperature of the nests (Kamel 2013; Kamel and

Mrosovsky 2006), the percentages of nests that would

have temperatures close to the PT (shaded nests) and nests

that would have feminizing temperatures (sun-exposed

nests and those under LV) were estimated according to

the thermal zones (HV, LV, and WV) along each beach.

The thermal zones of the beaches were categorized based

on the width (distance from the coastline to the HV

or limit where turtles could not nest, such as a human

infrastructure) and the dominant vegetation every 2 km

on Chenkan (18 km) and Punta Xen (18 km) beaches, and

every 5 km on Celesttn (22 km). These measurements

of the beaches, together with the geographical locations

recorded for the nests, were used to correlate them with

satellite images and the maximum distance of the nests

from the coastline (according to our results), through

Google images, in each beach per km (in linear m).

To document the degree of anthropogenic impact on

each beach, Google satellite images were used to measure

(in linear m) the disturbance along each beach, and

thereby estimate the percentage of human infrastructures

that would affect the nesting of the turtles per km. To

establish the number of nests per km throughout the

entire nesting period of the Chenkan and Punta Xen

beaches, databases were obtained from the Comision

Nacional de Areas Naturales Protegidas (CONANP)

and Grupo Ecologista Quelonios A.C., respectively.

The precipitation levels on the three beaches during the

entire nesting period were obtained from the Comision

Nacional del Agua (CONAGUA), in order to relate the

dates of both incubation periods with the amount of rain.

Statistical Analysis

The normality and homogeneity of the data were tested

by Kolmogorov-Smirnov and Levene’s tests. Wilcoxon

tests (as the distribution was not normal) were used to

compare the nest temperatures, hatching success, and

the variables that characterize the nests (incubation

April 2020 | Volume 14 | Number 1 | e233

Flores-Aguirre et al.

a) June-July August-September

i, S28 -

~ i A,

jae

- 32 - * J j j

wv s

5 df Locality

@ mR 7 ped Le fe EA ee = ss eed De ap ae Bee Seen [ereerao Weer? * ae J

e #8 a | : || Chenkan

ab] =

F 50 4 .

ao * *

* : Lal Punta Xen

Celestun

b) Pivotal temperature

Females pig

Nests by Locality

(29.45° C)

Fig. 2. (a) Incubation temperatures of Eretmochelys imbricata, during the TSP, on the beaches of Chenkan, Punta Xen, and

Celestun, during two incubation periods (June—July and August-September). The dotted line indicates the pivotal temperature. (b)

Percentages of females in the monitored nests on the nesting beaches of the western Yucatan Peninsula (Mexico). The red asterisks

show where the differences were significant (p < 0.05) among nests, by beach.

chamber temperature, depth, distance to the vegetation

and to the coastline, as well as percentage of shade

above the nests) between the two incubation periods

(June-July and August-September) within each beach.

An ANOVA was performed to compare the different

variables that characterize the nests among the three

monitored beaches. To determine the TSP of each nest,

the days from the beginning to the end of incubation

were calculated, and the middle third of the incubation

period was established. Subsequently, the temperature

during the TSP was compared among the beaches by a

Generalized Linear Model (GLM), with the set of nest

temperatures at each beach as the response variable. To

compare the temperatures during the TSP among the nests

on each beach, a GLM was used, with the temperatures

recorded for each nest as the response variable. In

addition, a time series was used, where nest temperatures

were correlated with the time at which they were taken,

and were compared among all the nests for each locality.

Measuring these correlations established the time

required for shaded nests to absorb heat, in comparison

to sun-exposed nests. Statistical significance was set at

p < 9.05; and SPSS, Stata SE11, and R version 3.5.1 (R

Core Team 2018) were used for statistical analyses.

Results

During the 2017 nesting season, 20 nests were

characterized on Chenkan beach (10 in June and 10 in

August), 20 on Punta Xen beach (10 in June and 10 in

Amphib. Reptile Conserv.

193

August), and 12 on Celesttn beach (eight in June and

four in August). The nests characterized along the three

monitored beaches showed no significant differences in

the temperature of the incubation chamber, distance to

the coastline, or percentage of shade (F’, = 1.614, p =

0.214). However, the distance to the nearest vegetation

differed significantly among beaches, with nests in

Celestun being the farthest from vegetation (F, = -3.514,

p = 0.042). Likewise, nest depth differed significantly

among the beaches, with the nests on Chenkan beach

being the most shallow (F’, = 3.85, p = 0.036; Table 1).

The average temperatures of the monitored nests did

not vary significantly among the three beaches during

June-July (F, = 0.202, p = 0.819), nor in August—

September (F', = 0.462, p = 0.640, Table 2). The average

incubation temperature on Chenkan beach was higher in

August-September (31.89 + 1.36 °C) than in June—July

(30.78 + 1.08 °C, F,,,, = 2.246, p = 0.045). In Punta

Xen, only the average temperature during the TSP was

higher in August-September (31.386 + 1.164 °C) than

in June-July (29.869 + 0.728 °C, F,,,, = 2.857, p =

0.017; Table 2). The incubation temperatures across all

beaches and the entire nesting season were within the

thermal interval that allows embryonic development

(28.5 to 32.9 °C, Table 2, Fig. 2). The nest temperatures

recorded during the TSP of June-July at Chenkan beach

showed significant differences among them, according to

their locations (F, = 34.830, p < 0.001), and the same

was found at Punta Xen beach (F, = 49.911, p < 0.001).

For the nests that were found under the shade, more than

April 2020 | Volume 14 | Number 1 | e233

Feminizaton of Eretmochelys imbricata in Mexico

Table 1. Nest characteristics of Hawksbill Turtles in the western Yucatan Peninsula, Mexico.

Locality

Distance to

vegetation

Distance to

coastline (m)

Percentage of

shade (%)

)

Tavs [2938-194 [26.002254[44752 7.18] 25414851 [0490148 | 15064243 [ 9572 1912

TL aust} 2831.70 [27572115 [46452756] 28654781 [1.784352 | 19780653 [24442 3468

TL august [2878-046 [as9e3 1. 27]4o672 402] 35334751 [2454097 [2048553 | 225% 1061

70% of their total coverage throughout the incubation

period had significantly lower temperatures compared

to sun-exposed nests (p < 0.001, Fig. 2a). On Celestun

beach, the nest temperatures did not show significant

differences between these two conditions of shaded

(with less than 50% of total coverage) and sun-exposed.

The nest temperatures in the TSP of August-September

at Chenkan beach showed significant differences only

in one nest, with lower temperatures compared to the

other nests (F_ = 11.320, p< 0.001), and the same pattern

occurred on Punta Xen beach (F, = 13.493, p < 0.001).

Both of these nests on each beach were sun-exposed,

therefore they remained above the PT (Fig. 2a). The nests

that were found under total shade coverage throughout

the incubation period, exhibited a temperature damping

effect, taking 3.5-4 h longer to heat up to the same

temperature as the sun-exposed nests (p < 0.001).

Although the shaded nests had lower temperatures than

those that were sun-exposed, most of the nests were

exposed; and therefore, in general, the nest temperature

during the TSP was hotter than the PT.

The hatching success for June—July did not differ

significantly between the three beaches: Chenkan (61.9

+ 26.71%), Punta Xen (57.97 + 41.04%), and Celestun

(71.38 + 24.53%, F, = 0.54, p = 0.589). A similar trend

was observed in the period of August-September on

Chenkan (50.89 + 24.36%), Punta Xen (26.02 + 36.3%),

and Celestun (61.95 + 41.94%, F, = 2.30, p = 0.125),

although numerically lower hatching success was

observed in the nests located in Punta Xen (Fig. 3).

During the cleaning of the nests in both nesting

periods, 170 dead hatchlings were collected from 17 nests

(five from Chenkan, five from Punta Xen, and seven from

Celestun). In a nest from Chenkan beach (June—July), a

sample of 31 dead hatchlings was obtained, which were

predated by ghost crabs. In addition, on Punta Xen beach

(August-September), a sample of 65 dead hatchlings

was obtained, which were predated by ants (Solenopsis

germinata and Labidus coecus). From these two nests,

the largest and freshest samples were obtained because

the hatchlings were found less than 12 h after death.

In five nests, the hatchlings required more than 36 h

to emerge from the nests, therefore the tissues of dead

hatchlings were degraded and the sexes of 25 samples

were unidentifiable. In total, 145 gonads from 12 nests

were identified. Histological examination showed that

the ovaries of hatchlings exhibited a thickened cortex

and unorganized medulla, whereas testes presented a

very thin cortex and had an organized medullary region

with developing seminiferous cords (Fig. 4). From the

total sample, 126 females and 19 males were determined.

Only hatchling females were found in the sun-exposed

nests, with temperatures of 30.21 °C and above.

However, both sexes were found in the shaded nests,

with temperatures in the range of 28.55—29.59 °C (Table

3). The temperature variation in the nests during the TSP,

from which the histological samples were obtained, was

0.77 + 0.15 °C. These results were used as the input and

for calibration of the TSD software, which determined the

pivotal temperature as 29.45 (+ 0.147 SE) °C, according

to the maximum-likelihood analysis, with the Richard

model (Bae < 0.001, AIC = 61.54, Fig. 5). In addition,

the transitional range of the temperature was -0.53 +

0.048. According to the TSD software, only 27.02% of

the hatchlings were males in the monitored nests. The

highest percentage of females occurred during the late

nesting season (August-September, 83.33 + 4.01%) and

the lowest percentage occurred during the middle (June—

July, 65.49 + 4.08%, t,,, = -3.72, p = 0.006; Fig. 2b).

In the three studied beaches, the dominant vegetation

was the coastal dune halophyte, which is mainly low

and medium-sized, characterized by herbaceous plants

Table 2. Average nest temperatures during the total incubation period and the TSP of the Hawksbill Turtles for each monitored

beach, between the months indicated. Significant differences (p < 0.05) are indicated by asterisks.

SO Temperature during incubation (°C) Temperature during TSP (°C)

June—July August-September June—July August-September

30.78 + 1.08% 31.9 + 1.36% 30.49 + 0.72* 31.71 £0.89"

30.44 41.44 3136 £1.23 29.87 + 0.62* 31.63 40.68"

30.85 + 1.48 30.440.95 30.29 + 0.83 30.1+0.63

Amphib. Reptile Conserv. 194 April 2020 | Volume 14 | Number 1 | e233

Flores-Aguirre et al.

June-July

100 -

Hatching Percentage

Chenkan Punta Xen ~~‘ Celestuin

Locality

Chenkan

August-September

Locality

Chenkan

= Punta Xen

= Celestun

Punta Xen Celestun

Fig. 3. Hatching success of Eretmochelys imbricata on the beaches of Chenkan, Punta Xen, and Celestin, Yucatan Peninsula

(Mexico).

and shrubs. However, the high vegetation (HV; 1.e., the

vegetation that provides more shade) changed among the

three beaches. In Chenkan, the HV was dominated by

spots of coconut plantations, present only in 22.2% of

the total length of the beach, because natural vegetation

(mangroves) was located on the other side of a road. In

Punta Xen, the dominant HV was the mangroves, present

in 62.5% of the total length of the beach; whereas in

Celesttn, the HV was dominated by tropical dry forest,

which corresponded to 77.3% of the total length of the

beach. Nevertheless, the percentage of shade provided by

the HV to the nests was not equal to the percentage of HV

along the three beaches studied, since it depended on the

distance between the HV, the nests, and the coastline, as

well as the degree of anthropogenic disturbance. Thus,

according to the distance from the coastline to the HV

and the maximum distance between the monitored nests

and the coastline (30 m; Table 1) the percentage of the

shaded nest was calculated. Chenkan beach had the

lowest percentage of shaded nests, with only 2.89%, next

was the Celesttn beach with 21.54%, and the highest

was the Punta Xen beach with 29.59%. The percentage

of shade above the monitored nests on the three beaches

also changed between the periods of June-July and

August-September. In June-July, 50% of the nests on

Celestin were recorded under shrubs, 50% of the nests

in Punta Xen were under the shade of the mangroves, and

40% of the nests in Chenkan were under shade (coconuts

and man-made structures to create shade on the beach,

known as palapas). In contrast, in August-September,

most of the nests were found in less shaded areas: in

Celestun, 0% of the monitored nests were under shade,

in Punta Xen, 20% nests were shaded, and in Chenkan,

10% of nests were shaded.

According to the analysis of anthropogenic impacts,

Chenkan beach exhibited a 100% disturbance, mainly

due to the road along the beach. On average, the distance

was 17.38 + 7.55 m from the coastline to the road in the

most eroded area, which caused the nesting of the turtles

to be interrupted. In contrast, the wider area from the road

to the coastline had an amplitude of 140.57 m, however,

it had high deforestation and human infrastructures, such

Table 3. Numbers of dead hatchlings and percentages of females, according to the histological sex, temperature during the TSP at

which nests were incubated in situ, and the percentage of shade above the nests, by locality and incubation period.

Beach

August—

September

Amphib. Reptile Conserv.

; Temperature

t) it)

Shade (“%) No. dead hatchlings during TSP (°C) Females (%)

pO =e 28.55 + 0.66

PuntaXen [| 90 82884 £0.51

June-July SS as aes es eee

a [a (ee

a a aa aed |

ae | |

ae Se | ne

29.59 + 0.52

30.66 + 0.44

AT +0.

po S847 £0.73

| Pmaxen | /-—— 32.41 + 0.89

April 2020 | Volume 14 | Number 1 | e233

Feminizaton of Eretmochelys imbricata in Mexico

Bases eS Se eee

Fig. 4. Representative histology sections

(100 ym), (b)

Ovary (100 um), (c) Testis (20 um), (d) Millerian ducts of male (20 um), (e) Ovary (20 um), (f) Millerian ducts of female (20 um).

The dotted areas in (a) and (b) indicate the sites of the higher magnifications shown in (c) and (e), where the dotted line indicates

the cortex, M indicates the medulla region, and C indicates the cortex region.

as summer houses, restaurants, palapas, and fishing

areas. In addition, there were breakwaters in the sea.

Punta Xen beach had an average width of 22.86 + 11.55

m. The disturbance represented 14.95% of the length

of the beach, mainly in places which were recreation

areas, aS well as areas with high deforestation rates and

the presence of infrastructures, such as a hotel complex.

Celestun was the most well-preserved beach, presenting

only 6.03% disturbance, mainly due to the presence of

two hotel complexes and some summer houses. The

average width of the beach was 42.59 + 6.69 m.

According to the results, the percentage of shade

above the nests was the main factor that decreased

their temperatures, approaching a PT. Therefore, the

percentage of shaded nests, the degree of conservation of

the beaches, and the number of nests per km were used

to estimate the percentage of nests with temperatures

close to the pitoval for the 2017 nesting season. Chenkan

beach had the greatest feminization among all the nests,

showing only 0.5% of shaded nests. Punta Xen beach

had the highest percentage of HV near the nesting areas,

so it is estimated that 39.54% of the nests would have

temperatures close to the pivotal (shaded nests). For

Celestun beach, the HV was far from the nesting areas,

so only 21.54% of the nests would have been under the

shade.

With respect to the average rainfall, it differed

between the two incubation periods on the beaches of

Punta Xen and Chenkan. For June—July it was 10.61

+ 4.66 mm, while for August-September it was 7.22 +

2.68 mm. In Celestun, precipitation was very low during

both periods of incubation, without even reaching 1 mm

Amphib. Reptile Conserv.

of precipitation, while the relative humidity remained

between 74% and 76% for all incubation months,

increasing to 95% in September.

Discussion

In species with genetic sex determination systems,

Fisher's theory predicts that the sex ratio in a population

tends toward 1:1 (males:females). Given that when either

sex 1s in the majority, the other sex has an advantage

in finding mates, an equal sex ratio represents the

evolutionarily stable strategy. However, in organisms

with temperature-based sex determination this balance

can become biased by warm or cold environments

(Janzen and Phillips 2006).

The beaches have different environmental

temperatures, which provide diverse thermal gradients

to the nests (Diffenbaugh and Field 2013; Hawkes et

al. 2009; Marcovaldi et al. 2014), so differences in

incubation temperatures among nesting beaches would be

expected. In the present study, despite the differences in

littoral characteristics and conservation status among the

three monitored beaches, and the distinct geographical

position of Celestun relative to the other two beaches, no

significant differences in incubation temperatures were

found. This result may be due to the nesting characteristics

of the Hawksbill Turtles, since most nests were found

in the dune zone, which is associated with vegetation

(Horrocks and Scott 1991), and nest temperatures are

influenced by the type of vegetation present in each

beach. Nevertheless, the low vegetation (LV) and the

edge of the high vegetation (HV) do not provide lower

April 2020 | Volume 14 | Number 1 | e233

Flores-Aguirre et al.

Pivotal temperature = 29.45 °C

ras)

ran)

_—

re)

vo oO

i48)]

var

2 8

oa.

v 2

o wv

> oe

rot]

29 30

Oo 90 0@ 00 OOM 00 O@M Oo

31 32 33

Temperature (°C)

Fig. 5. Relationship between temperature and sex ratio. The red dotted line denotes the pivotal temperature according to the TSD

software and calibrated with histology of gonads from dead Hawksbill hatchlings.

temperatures, and only the direct influence of the HV

promotes lower temperatures and therefore more male

hatchlings (Kamel 2013; Kamel and Mrosovsky 2006).

On all three beaches, the vegetation in the nesting areas

was low, which failed to decrease the temperatures. Also,

the HV near the nests was scarce (less than 30% of the

total nests by locality), which caused the majority of the

nests to exhibit similar thermal conditions. However,

the conditions of each beach differed in the factors

that determine the nest temperatures. On Chenkan

beach, the least HV occurred because of the high

level of anthropogenic impact, primarily by the road

that divided the nesting area from the native HV. As a

consequence, nesting was prevented in the areas with

lower temperatures (below the mangrove), which caused

the majority of the nests to be exposed to feminizing

temperatures. In contrast, although Celestin was the

most well-preserved beach, the HV was generally found

inland from the coastal dune, so the distance between

the zone of HV and the coastline was wide. On all three

beaches, the distance from the nests to the coastline was

an average of ~17.83 m, and most turtles nested in the

dunes. As a consequence, few turtles reached the zone of

HV, which was located at an average of ~45.59 m from

the coastline. Therefore, the majority of turtles nested in

areas near LV, so most of the nests would have been in

feminizing temperatures. Punta Xen beach had patches

with HV (mangroves), so it was the beach with the

highest percentage of shaded nests. Thus, the incubation

temperatures among the nests were diverse, generating

the highest percentage of nests close to the PT among

the three monitored beaches. However, in this camp the

nests are routinely translocated to hatcheries, due to high

predation, so it is not known how this action would affect

the sex ratio of the hatchlings. Therefore, despite the

differences in the beaches, the trend toward feminization

of Hawksbill Turtle hatchlings remained. Unfortunately,

in the western Yucatan peninsula, the environmental

temperatures are warmer now than 40 years ago (Sinervo

Amphib. Reptile Conserv.

et al. 2010), and they also are in the studied beaches

(~2.33 °C; CONAGUA). Even though vegetation can

play an important role in reducing the nest temperatures,

as environmental temperatures continue to rise, the trend

toward feminization of hatchlings will increase.

Sea turtles lay several nests over a nesting season, so

the environmental temperatures trigger differences in the

sex ratio, with a higher percentage of female hatchlings

produced in the middle of the nesting period (Mrosovsky

et al. 1984). The seasonality in our study area also

modified the nest temperatures and, therefore, the sex

ratio of the hatchlings. However, contrary to the study of

Mrosovsky et al. (1984), we found higher temperatures

(~31.89 °C) at the end of the nesting season (August—

September) than during the middle (June—July, ~30.78

°C). These lower temperatures were due to the early rains

(June-July) that reduced the nesting temperature and

increased the proportion of male hatchlings (Wyneken

and Lolavar 2015). In addition, the rain causes an increase

in foliage, so most of the nests in June—July were shaded,

which influenced the temperature difference between the

two incubation periods.

Hatching success did not vary significantly among the

beaches and months studied. In contrast, Pike (2008, 2009)

previously found that beaches with greater anthropogenic

influences have reduced hatching success compared to

better preserved beaches. Although we did not observe

a change in the percentage of hatching among beaches,

the degree of conservation did influence the survival

of the nesting females. Particularly at Chenkan beach,

several nesting females (six) were run-over by cars while

trying to cross the road in order to nest. Nest predation

was the main factor that influenced hatching success in

the three studied beaches. As this factor was similar on

all three beaches, the recruitment of hatchlings of both

sexes to the sea did not change. However, on Punta Xen

beach it was possible to observe greater predation of the

nests left in situ, which could be related to the shortest

distance between the mangrove and the beach, as well

April 2020 | Volume 14 | Number 1 | e233

Feminizaton of Eretmochelys imbricata in Mexico

as that beach’s low level of anthropogenic impact, which

allowed natural predators of turtle eggs to gain greater

access to the nests.

According to the temperatures recorded in all the

monitored nests, incubation temperatures did not

approach the thermal limits that prevent embryonic

development (25—34 °C; Howard et al. 2014). The lowest

temperature recorded in the nests was 28.55 °C, while

the highest temperature was 32.85 °C, which produced

hatching successes of 83.84% and 81.54%, respectively.

Therefore, 1n these locations, the incubation temperatures

are still maintained within the optimum temperatures for

the embryonic development of Hawksbill Turtles.

Based on gonad histology and the temperatures

recorded during the TSP of nests in situ, the feminizing

temperatures (30.21 °C and above) were determined, as

well as the temperature range that produces both sexes

(28.55—29.59 °C) for this population. These temperatures

are similar to those reported for other populations of

Hawksbill Turtles, including those obtained under

controlled incubations. This is because the depth of

the nests provides a buffer that prevents environmental

fluctuations (Booth 2006), and the temperatures of nests

in situ show minimal variation (+ 0.07 °C) during the

TSP, remaining close to the temperature variations in

experiments under laboratory conditions (+ 0.05 °C;

Mrosovsky et al. 2009). Therefore, these results suggest

that the use of dead hatchlings from nests in situ is a

reliable way to estimate sex ratios among hatchling sea

turtles.

According to controlled experiments in Australian

Hawksbill populations, temperatures below 28.4 °C

were masculinizing, while temperatures between 28.9

and 29.8 °C produced both sexes, and the temperatures

that only produced females were 30.4 °C and above

(Loop et al. 1995). In this study, no nests were found

with exclusively masculinizing temperatures, since

the lowest temperature registered during the TSP was

28.55 °C, from which most of the samples were males

(83.33%), but some females were still found (16.67%).

Likewise, in 1997, a study in Bahia, Brazil, found the

feminizing temperature was 30.4 °C and above (Godfrey

et al. 1999). However, in 1998, a study in Milman,

Australia, found temperatures of 28 °C and below were

masculinizing, whereas between 29.5 and 31 °C both

sexes were produced, and at 32.5 °C only females were

found (Dobbs et al. 2010). Those studies showed higher

pivotal and feminizing temperatures compared to the

present study. In general, the temperatures recorded in

previous studies were similar to the results here, despite

the geographical distances, as well as the differences in

time.

The pivotal temperature (PT), determined according to

the histological results and the TSD software, was 29.45

°C, which is close to the PT interval of other studies,

even for different species, locations, and years (Hawkes

et al. 2009). Specifically for £. imbricata, in controlled

Amphib. Reptile Conserv.

incubations on Antigua in 1989, the PT was found to be

29.2 °C (Mrosovsky et al. 1992). Similar results were

found on Milman Island, Australia, in 1998 (Dobbs

et al. 2010); and in Bahia, Brazil, in 1997, the PT was

reported as 29.66 °C (Godfrey et al. 1999). Therefore,

it appears that the PT has remained relatively constant

over the years and in different regions, which supports

the hypothesis that this trait is highly conserved in sea

turtles (Hawkes et al. 2009; Mrosovsky et al. 1992). This

constancy makes it unlikely that the PT in sea turtles will

evolve substantially in response to climate change events,

which are occurring very rapidly on an evolutionary

timescale (Hawkes et al. 2009), and are thus leading to

feminization in many populations.

This study highlights the trend towards the feminization

of the Hawksbill Turtles, at least in the western Yucatan

Peninsula during 2017, since 82.05% of the monitored

nests produced only female hatchlings. This trend toward

increased female production was present in all monitored

beaches, most remarkably in Chenkan. This feminization

trend of Hawksbill Turtles has already been reported in

previous studies. For example, from 1991 to 1997, in

Bahia, Brazil, the sex ratio was estimated according to

the incubation duration. Across six nesting seasons, the

percentage was greater than 90% females (Godfrey et

al. 1999). Wibbels et al. (1999) also observed a strong

tendency toward feminization on Buck Island. Based on

gonad histology of dead hatchlings in 51 nests in situ, 49

nests produced only females, while in the two remaining

nests both sexes were found. Likewise, Glen and

Mrosovsky (2004) reported changes in the sex ratio of

Hawksbill hatchlings in Antigua, relative to those found

in the work of Mrosovsky et al. (1992), which found in

1989 that sand temperatures were below the PT. When

repeating this earlier work in 2003, Glen and Mrosovsky

(2004) reported an increase in air temperature which

increased the temperature for most of the nests above

the PT. So, they proposed that in the face of climate

change, the ratio of males will be reduced. In Bahia

and Rio Grande do Norte, Brazil, data from 27 years of

incubation durations found that 89-96% of the hatchlings

were females in both locations (Marcovaldi et al. 2014).

This trend towards feminization has been documented

mainly in hatchlings. However, Hawkes et al. (2013)

made a summary of the sex ratios of Hawksbill Turtles

and found that the bias toward feminization is greater in

hatchlings than in young and adult turtles, suggesting

that 1t may be due to high predation during this stage.

Nevertheless, in a study in Anegada, a site characterized

by an important foraging aggregation of Hawksbill

Turtles from different areas of the Caribbean, Hawkes

et al. (2013) measured the levels of testosterone and

oestradiol in the blood of juvenile Hawksbill Turtles,

and found 2.4- to 7.7-fold more females than males. So,

between 69% and 89% of juvenile Hawksbill Turtles

were females, which also shows a greater feminization

during the juvenile stage. In adults and sub-adults, the sex

April 2020 | Volume 14 | Number 1 | e233

Flores-Aguirre et al.

ratio of Hawksbill Turtles in Cuba in 1985 and 1986, was

77% females (Carrillo et al. 1998). Therefore, according

to the reviewed studies, the feminization trend has been

maintained for all life stages of Hawksbill Turtles in the

Caribbean.

The Hawksbill Turtle population of the Yucatan

Peninsula exhibits high philopatry to the nesting and

breeding areas (Cuevas et al. 2012; Labastida-Estrada

et al. 2019) among both females and males (Gonzalez-

Garza et al. 2015). The bias towards feminization can

thus increase their vulnerability to inbreeding due to

decreased gene flow (Hudson 1998). This trend may be

exacerbated if the connection among the populations is

lost due to habitat loss and fragmentation (Witherington

et al. 2011), which could cause a reduction in population

sizes, decreasing their genetic variability and increasing

the risk of extinction (Gonzalez-Garza et al. 2015;

Spielman et al. 2004).

In the Yucatan Peninsula, two management units

(subpopulations) are established in the Hawksbill Turtles,

according to their haplotype composition. Genetically,

the turtles segregate into two populations: one in the

Gulf of Mexico (west of the Yucatan Peninsula) and one

in Yucatan and Quintana Roo (northeast of the Yucatan

Peninsula; Abreu-Grobois et al. 2003; Labastida-

Estrada et al. 2019). Gonzalez-Garza et al. (2015) found

differences in the rates of multiple paternity between

these two subpopulations of Hawksbill Turtles, with

only the hatchlings from north of the Yucatan Peninsula

exhibiting multiple paternity. The lack of multiple

paternity among hatchlings from the Gulf of Mexico,

may be a consequence of reduced mate availability

(Bowen and Karl 2007; Tedeschi et al. 2014).

Hawksbill populations of the Yucatan Peninsula

contain endemic haplotypes (Labastida-Estrada et al.

2019), which can be explained by historical patterns of

gene flow among populations (Reece et al. 2005). The

population of the Yucatan Peninsula is genetically isolated

from other populations of the Caribbean, because gene

flow decreases across the Caribbean populations towards

the Yucatan, due to the emergence of the Campeche bank

and the Florida Shelf during the Pleistocene (Reece et

al. 2005). At present, in Hawksbill Turtle populations on

the Yucatan Peninsula, a lower genetic variability has

already been documented in the hatchlings of females

that nested for the first time (neophytes) as compared to

the hatchlings of remigrant females, which suggests a

loss of genetic variation through time (Gonzalez-Garza

et al. 2015). This trend may be exacerbated by the bias

toward feminization in this population of Hawksbill

Turtles, as revealed in this study.

The bias towards feminization could also be further

driven by the effect of climate change, since an increase

in temperature even of 1—2 °C, can have considerable

effects on the sex ratio (Janzen 1994). In the region

of Mexico and Central America, it is estimated that

environmental temperatures will exhibit an increase in

thermal anomalies of 1.5 to 5 °C, due to the decrease in

Amphib. Reptile Conserv.

precipitation between 2,000 and 2,100 mm (Christensen

et al. 2007). Therefore, feminization would increase,

limiting the possibility of adaptation and leading

to population declines in species that are already in

danger of extinction (Hamann et al. 2010; Hawkes

et al. 2007; Mitchell and Janzen 2010). In addition,

climate change accelerates beach erosion processes,

which currently have an estimated erosion rate of 1.8

to 6.8 m/yr for the state of Campeche (Botello et al.

2010). This is driven by human infrastructure, which

is the main cause of the destruction of coastal beaches

(Botello et al. 2010), as well as the high deforestation

that exists in the coastal dunes, severely affecting

the nesting and the sex ratio of Hawksbill Turtles, so

measures must be taken to reduce it.

Conservation Implications

The trend toward feminization of Hawksbill Turtles

has now been documented in several studies, including

this one. This bias is predicted to increase in the face of

climate change, severely affecting Hawksbill populations,

which are already Critically Endangered. Therefore,

according to our results, we propose the establishment

of conservation strategies that promote balanced sex

ratios in these turtles. Since the shade provided by the

vegetation is a natural component that decreases the

nest temperatures, and therefore maintains the sex ratio

of Hawksbill hatchlings, priority should be given to

vegetation preservation on the nesting beaches, as well

as the realization of reforestation programs, especially of

the native vegetation that provides greater coverage to

the nests. In addition, vegetation is vital for the survival

of a wide variety of other species and for maintaining the

stability and integrity of the beach. In some turtle camps

in Mexico, as 1s the case of Punta Xen, the conservation

strategy is to move the nests in situ to hatcheries, to reduce

their losses due to predation. Therefore, we recommend

keeping nests in situ, in areas with greater vegetation

cover, and carrying out continuous monitoring to reduce

the predation. Or, where appropriate, we recommend the

use of at least 50% artificial shade in the hatcheries, in

order to provide more equitable recruitment of hatchlings

in terms of their sex ratio. These measures should continue

to promote the recovery of Hawksbill Turtle populations,

and ensure their viability in the face of climate change.

Acknowledgments.—We thank the Posgrado en Ciencias

Biologicas, UNAM, and CONACYyT for the scholarship

awarded to Cynthia Dinorah Flores-Aguirre (CVU

545214); and UNAM PAPIIT/DGAPA (IN201218;

IN210116; IN212119) for financial support; as well

as CONANP, Grupo Ecologista Quelonios A.C., and

PRO NATURA Peninsula de Yucatan A.C., for all their

support for lodging 1n the camps and the use of ATVs. In

addition, thanks to all the personnel of the three camps

for their great collaboration during the fieldwork. We

also thank CONANP and Grupo Ecologista Quelonios

April 2020 | Volume 14 | Number 1 | e233

Feminizaton of Eretmochelys imbricata in Mexico

A.C. for sharing their data of the nesting of Hawksbill

Turtles in 2017, especially to Mr. Javier Cosgalla for all

his support in carrying out this study. As well, we thank

to Coordinacién General del Servicio Meteoroldgico

Nacional (CGSMN) de la Comisién Nacional del Agua

(CONAGUA) for providing us with the environmental

temperature and precipitation data of the study areas.

Likewise, we thank the Biol. M. A. Suastes Jiménez for

the identification of the ants that preyed on the nests.

This article is a requirement for obtaining the degree

of Doctora en Ciencias Biologicas del Posgrado en

Ciencias Bioldgicas, UNAM, for C.D. Flores-Aguirre.

The manuscript was improved by the comments of

the reviewers Labastida-Estrada, V. Guzman and an

anonymous reviewer, to whom we are grateful; especially

to V. Guzman, who helped us obtain relevant information

from the turtle camps in which we had the pleasure of

working. Also, the manuscript has been improved by E.

Bastiaans for a helpful review of the English language.

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Cynthia Dinorah Flores-Aguirre is a Ph.D. candidate in the postgraduate program in Biological Sciences

at the Universidad Nacional Autonoma de México (UNAM). Her main interests are the conservation,

ecology, and behavior of various fauna, as well as their vulnerability to climate change, focusing on the

Ver6nica Diaz-Hernandez is a Biologist who received her Ph.D. in Biomedical Sciences from the

Universidad Nacional Autonoma de México (UNAM) in 2008. Veronica is currently a Full Professor in

the Department of Embryology at UNAM. Her main interests are in sexual determination among reptiles,

and more recently in the vulnerability of reptiles to climate change.

Isaias Hazarmabeth Salgado Ugarte is a senior Full Professor at Laboratorio de Biometria y Biologia

Pesquera, Facultad de Estudios Superiores Zaragoza Campus, Universidad Nacional Autonoma de México

(UNAM). Isaias earned his Bachelor of Science in Biology, and his Master of Science in Biological

Sciences from UNAM; and he obtained a Fisheries Biology Specialty and a Ph.D. in Aquatic Biosciences

(Fisheries Biology) from University of Tokyo, Japan. He has been working on exploratory, confirmatory,

univariate, bivariate, multivariate, computer intensive statistical methods (nonparametric kernel density

and regression smoothing, cross-validation, bootstrapping) to analyze biological data.

Luis Enrique Sosa Caballero is a Biologist who graduated from the Facultad de Ciencias, Universidad

Nacional Autonoma de México in 2015. Luis completed his Master of Science in the Institute of Biology

of the same university, obtaining the degree with honorable mention in 2019. His main topics of interest

are conservation, climate change, and thermoregulation, focusing on groups of amphibians and reptiles,

with special attention on reptile species which have sex-determination by temperature, such as sea turtles

and crocodiles. In addition, he has an interest in the geographical distributions of different groups under

predictive scenarios and climate change.

Fausto R. Méndez de la Cruz is a Full Professor at the Universidad Nacional Autonoma de México

(UNAM) in México City. Fausto obtained his Ph.D. in 1989 from UNAM and did postdoctoral studies

at the University of Florida, Gainesville, Florida, USA, and a sabbatical at Virginia Polytechnic Institute

and State University, Blacksburg, Virginia, USA. His research is focused on the evolution of reproductive

systems in reptiles, mainly on the evolution of viviparity and parthenogenesis, and more recently on the

vulnerability of amphibians and reptiles to climate change.

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