Official journal website:

amphibian-reptile-conservation.org

Amphibian & Reptile Conservation

16(1) [General Section]: 1-13 (e298).

The anuran fauna in a protected West African rainforest and

surrounding agricultural systems

‘Konan Hervé Oussou, '*’N’Guessan Emmanuel Assemian, ‘Atta Léonard Kouadio,

2Manouhin Roland Tiédoué, and *Mark-Oliver R6édel

'Jean Lorougnon Guédé University, Laboratory of Biology and Tropical Ecology, UFR Environnement, BP 150 Daloa, COTE D’IVOIRE ?Ivorian

Office of Parks et Reserves, OIPR, BP 1342 Soubré, COTE D’IVOIRE 3>Museum fiir Naturkunde, Leibniz Institute for Evolution and Biodiversity

Science, Invalidenstrasse 43, 10115 Berlin, GERMANY

Abstract—The conversion of tropical rain forests to agricultural systems is a major threat to tropical

biodiversity. In West Africa, studies investigating the effects of this habitat conversion on biodiversity are

scarce. In this study, we investigated which forest amphibians survive in the agroforestry systems surrounding

West Africa’s largest area of protected rainforest, the Tai National Park (TNP) in south-western Cote d’lvoire.

Species richness was assessed in different habitats types, i.e., a mosaic of coffee and cocoa plantations,

rubber plantations, and rice fields, and compared to data from primary and degraded forests in TNP. The

anuran assemblage composition differed considerably between forest and agroforestry systems, with the

latter comprising only a small subset of generalist forest species and species which usually occur in highly

degraded forest habitats or even savanna. Thus, the agroforestry systems in western Cote d’lvoire seem to be

unsuitable for the maintenance of the rich local and regional diversity of forest amphibians.

Keywords. Agroforestry, amphibians, biodiversity, habitat conversion, Ivory Coast, Upper Guinea forest zone

Citation: Oussou KH, Assemian NE, Kouadio AL, Tiédoué MR, Rédel M-O. 2022. The anuran fauna in a protected West African rainforest and

surrounding agricultural systems. Amphibian & Reptile Conservation 16(1) [General Section]: 1-13 (e298).

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.

Accepted: 10 December 2021; Published: 14 January 2022

Introduction

While tropical rainforests may comprise about half of the

world’s biodiversity, they are shrinking at a very high rate.

A comprehensive investigation on the African continent

revealed that between 1990 and 2000, over 50 million ha

of forest disappeared (FAO 2001, 2006). In West Africa,

the situation is especially alarming as only 20% of the

1.5 million km? of forest present at the beginning of the

20" century persists to date (FAO 2006). The main driver

of forest loss is logging and conversion into plantations

(mainly oil palm, cocoa, coffee, and rubber), and other

forms of agricultural land use.

Cote d’ Ivoire is no exception to this trend of increasing

forest loss. For example, Chatelain et al. (1996)

revealed that in the most forested part of the country,

the far south-west, 79% of forest had been lost in only

20 years. However, one large area of rainforest in that

region prevailed: the Tai National Park (TNP). At about

536,000 ha, TNP represents more than 50% of the total

area of protected West African rainforest (OIPR 2014),

thus constituting the most important area of protected

rainforest in the entirety of West Africa. However, the

Correspondence. “assemanuel@yahoo,fr, *>mo.roedel@mfn. berlin

Amphib. Reptile Conserv.

forests previously surrounding TNP, 1.e., various forest

fragments (Hillers et al. 2008) and classified forests

(Alonso et al. 2005), are now almost completely logged or

converted to small scale agricultural areas and industrial

sized plantations (Rédel et al. 2021; all authors, pers.

obs. ).

The amphibian fauna of TNP and the surrounding

classified and fragmented forests have been the focus of

numerous taxonomic and ecological studies (Perret 1988;

Rodel 1998; Rédel and Ernst 2000, 2001a,b, 2002a,b,

2004; Rodel et al. 2001, 2002a,b, 2003, 2004; Rodel

and Branch 2002; Veith et al. 2004; Rudolf and Rodel

2005, 2007; Sandberger et al. 2010; Kpan et al. 2019,

2021). These studies have revealed that the amphibian

communities show considerable differences between old

growth and previously selectively logged forests (Ernst

and Rodel 2005, 2006). Hillers et al. (2008) showed

that these differences were more strongly associated

with forest degradation rather than fragmentation. Ernst

et al. (2006) and Ernst and Rodel (2008) observed that

different frog groups react differently to logging, and

emphasized that although species richness may remain

similar between logged and pristine sites, functional

January 2022 | Volume 16 | Number 1 | e298

The anuran fauna in a protected West African rainforest

diversity dramatically declines in altered West African

rainforests. A recent study revealed that the recovery of

amphibian assemblages in previously logged parts of

TNP does occur, but it is very slow (Kpan et al. 2021).

All these studies collectively highlight the sensitivity of

forest amphibians to forest degradation, an observation

that was also made in other West African forests (Ofori-

Boateng et al. 2012; Adum et al. 2013).

However, we do not yet know to what extent West

African forest amphibians might be able to persist in

agroforestry systems. These artificial habitats now make

up almost all the areas surrounding TNP, providing

resources to local communities such as income, energy,

Shelter, and food (PNUE 2012). Studies from Asia

have shown that agricultural areas, which comprise

highly disturbed forests, may provide habitat for some

amphibian species and thus could be a component for the

maintenance of at least some part of the local biodiversity

(Wanger et al. 2009; Faruk et al. 2013; Konopik et al.

2015). However, respective studies from West Africa are

lacking so far.

This study aimed to determine the resistance of West

African forest amphibians to the conversions of their

original habitats to agricultural habitats. We examined

anuran species richness and species turnover in three

different habitats types, 1.e., mosaic coffee and cocoa

plantations, rubber plantations, and rice fields, and

compared them to the primary and degraded forest in

the TNP. We hypothesized that (a) species richness

decreases, and (b) species turnover increases, with

increasing disturbance between the habitat types.

Materials and Methods

Study area and study sites. Tai National Park is located

in south-western Cote d’Ivoire, between the Cavally and

Sassandra rivers, and the towns of Guiglo, Buyo, San

Pedro, and Tabou. It extends from 05°08’N to 06°24’N

latitude, and from 06°47’°W to 07°25’W longitude

(OIPR 2015). The climate of TNP is sub-equatorial with

four seasons: a long rainy season from mid-March to

July, a short dry season in August, a short rainy season

from September to October, and a long dry season

from November to mid-March. However, this regular

seasonality has changed to some extent in recent years.

The average annual rainfall is 1,800 mm, and ranges

from 1,700 mm in the north to 2,200 mm in the south

of the park (Chatelain et al. 2001). The average monthly

temperature varies from 24-28 °C (Koné 2004). The

relative air humidity is always high, ranging from 85%

to 90% under the forest cover, and usually reaches 100%

during the night (Bousquet 1978). The West African dry

season wind, called Harmattan, is irregular and has little

impact on the area, usually only extending over one to

two weeks in December or January (Adou et al. 2005).

Tai National Park is the largest prevailing rainforest

area in the Upper Guinea biodiversity hotspot (Myers et

Amphib. Reptile Conserv.

al. 2000; Bakarr et al. 2001). Botanically, TNP is part

of the large Guinean-Congolese floristic region (Dupuy

et al. 1999), and the flora of TNP comprises more than

1,350 species, of which 80 are endemic (Chatelain and

Kadjo 2000). The Park includes approximately 145

mammal species, corresponding to 93% of the mammal

fauna of the western Guinean forest zone (Riezebos et al.

1992), as well as 234 bird species (OIPR 2014), 60 fish

species (Grell et al. 2013), 43 snake species (Rodel and

Mahsberg 2000; Ernst and Rodel 2002), four turtles, two

crocodiles, and 11 lizard species (MOR, unpub. data).

So far 56 amphibian species have been recorded from

TNP (Rodel and Ernst 2004; Ernst et al. 2006), two of

which seem endemic; namely Phrynobatrachus taiensis

(Phrynobatrachidae; Perret 1988), and HAyperolius

nienokouensis (Hyperoliidae; Rodel 1998).

We surveyed the amphibians in different habitat types

in five sectors of TNP: ADK/V6, Soubré, Tai, Djapadj1,

and Dyjouroutou (Fig. 1). These habitat types were either

located inside TNP, along the park’s periphery, or in

agricultural systems surrounding TNP. The habitats

inside TNP consisted of two types: (1) primary forests

characterized by dense forests with a high, closed canopy,

and (relatively) open undergrowth, and (ii) degraded

forests with large canopy gaps and denser understory.

The degraded forests were mainly encountered at the

edge of the TNP, often in direct proximity to crops. All

agricultural sites where in the periphery of the TNP, and

consisted of a mosaic of coffee and cocoa plantations,

rubber plantations, and rice fields, often in close

06°24’N

—_— %

: awa nt

Legend

@ Sites

River te

@ Buyo lake @

Sectors >

MM ADK-V6

MS Djapadii

Djouroutou

MM soubré &

meal

7 al

Tai §

10 0 § 10 km

05°08’N | | |

dein Ww

Fig. 1. Locations of the 32 study plots in the Tai National Park

and surrounding agroforestry systems (see Appendix | for the

plot list and habitat descriptions). Inset figure: position of Tai

National Park in Céte d’ Ivoire.

07°28'W

January 2022 | Volume 16 | Number 1 | e298

Oussou et al.

Fig. 2. The different habitats in and around Tai National Park which were surveyed for amphibians. (A) and (B) near primary forest;

proximity to villages (Fig. 2). Rice fields usually have no

canopy and thus are not regarded as agroforestry habitats.

However, in the West African forest zone (and elsewhere,

compare Ndriantsoa et al. 2017), swamp forests are often

cleared in order to establish rice fields (R6del and Glos

2019). Therefore, we included rice fields here as well.

The geographic positions of all study sites were recorded

with a hand-held Garmin 60 CSx. A list of all sites,

including a brief description of the habitats, is included

in Appendix 1.

Amphib. Reptile Conserv.

Sampling methods. The fieldwork was conducted from

May to October 2018, thus covering the long and short

rainy seasons, as well as the short period of reduced

precipitation in August. A total of 32 plots of 100 x

50 m were established, 20 within the park and 12 in

the agroforestry systems (Figs. 1-2). The sites in TNP

comprised 14 plots in primary forest, and six in degraded

forest. In the TNP periphery, three plots were established

in coffee/cocoa plantations, two in rubber plantations,

and seven in rice fields. Each plot was visited twice (total

January 2022 | Volume 16 | Number 1 | e298

The anuran fauna in a protected West African rainforest

YOQOOQOOO

nQ00002

OOOO” OC

QQ0°

9°Ce

Number of species

I, 3) 5

=i Sobs =f (Chao ? O- Jack 1

DOQOOOODOOOOOO00D000000000 OO00

ee eS

7 9 1113 15 17 19 21 23 25 27 29 31 33 35 3/7 39 41 43 45 47 49 51 53 55 5/7 59 61 63

Sample (days)

Fig. 3. Species accumulation curve (triangles) and estimated amphibian species richness (Chao 2, squares; and Jack-knife 1, circles)

of the Tai National Park and surrounding agroforestry system. The mean values of 500 random runs of the daily species lists are

given. A daily species list comprised the presence/absence records collected during seven hours of sampling (four hours during

daylight, and three hours during night) on one plot by two people for a total of 14 person-hours.

64 days) with a consistent sampling intensity of 7 h/visit

(four hours during daylight at 0700-1100 h, and three

hours during night at 2000-2100 h GMT). Plots were

always investigated by two people (for 14 person-hours

per plot day), so the search effort represents a total of 896

person-hours of sampling. The average time between the

two visits to a plot was three months (see Appendix 1).

Amphibians were located through visual and acoustical

searches in all the different habitats of a plot (Heyer et al.

1994; Rodel and Ernst 2004). Individuals encountered were

identified to the species level following the nomenclature

of Frost (2020). It should be noted that Arthroleptis cf.

poecilonotus (Fig. 4D) comprises a complex of species,

which currently cannot be assigned with certainty to

a valid name (compare Roédel and Bangoura 2004;

Blackburn 2010; Blackburn et al. 2010; Channing and

Rodel 2019). In TNP, two species occur in syntopy, and

they are indistinguishable based on morphology and

habitat preferences but discernible by their calls (Ernst

and Rodel 2005). In this study, we treated them as a single

taxon. Representatives of each species were collected,

anesthetized in a chlorobutanol solution, and thereafter

preserved in 70% ethanol. These voucher specimens were

deposited in the collection of the Hydrobiology Unit of

the Laboratory of Biology and Tropical Ecology at Jean

Lorougnon Guédé University, Cote dIvoire.

Data analysis. As species detection probability seemed

to differ among the species, and numbers of calling

males could not be reliably counted on the plots, we used

qualitative data for the assessments but not quantitative

data. The estimated species richness, and thus the

sampling efficiency, were calculated with the Jack-

knife 1 and Chao 2 estimators using EstimateS software

(Version 9.1.0; Colwell 2013). These two estimators

Amphib. Reptile Conserv.

are incidence based. The presence/absence data of the

daily species lists (64 days of survey work) were used

for all 41 species recorded in the entire study area, as

well as for the different habitat types (see Table 1). To

avoid order effects, 500 random runs of the daily species

lists were conducted. In order to test for similarities or

differences in amphibian species compositions in the

investigated habitat types, the Jaccard’s Similarity Index

was calculated using the software PAST (Version 2.17c;

Hammer et al. 2001).

In order to determine whether a particular species

disappeared after habitat degradation, survived in the

agroforestry systems, or even invaded such habitats, we

classified each species in the following three categories

(bearing in mind that the total numbers may deviate from

the total number of recorded species as not all categories

were known for all species, compare Table 1):

I. Habitat: F = species only occurring in closed forest:

D = species occurring in degraded forest (may occur in

closed forest, absent in open areas); O = species living in

open areas (may occasionally be encountered in degraded

forest); if a species could be observed in more than one

category, it was assigned to the ‘most open’ habitat.

II. Micro-habitat of adults: f = fossorial; a = aquatic; |

= leaf litter; t = arboreal.

III: Micro-habitat of tadpoles: d = direct developers

and non-feeding non-hatching tadpoles, developing

in leaf litter or in the soil; s = stagnant waters (ponds,

puddles, tree holes); f = flowing water.

Results

Species richness and community composition. Overall,

the surveys revealed 41 anuran species from 12 families

January 2022 | Volume 16 | Number 1 | e298

Oussou et al.

Table 1. Amphibian species recorded in the Tai National Park and surrounding agricultural areas, along with recorded and estimated

Species numbers per habitat type and general habitat preferences of adults and tadpoles (see Materials and Methods). Abbreviations:

PF: primary forest; DF: degraded forest; CP: mosaic of coffee and cocoa plantations; RP: rubber plantation; RF: rice field; 1: leaf

litter; t: arboreal; a: aquatic; f: fossorial; F: closed forest; D: degraded forest; O: open habitat; ? : unclear as species identification

was not possible; d: direct development and non-hatching/non-feeding tadpoles; s: tadpoles in stagnant water (ponds, puddles, tree

holes); f: tadpoles in flowing water; *: comprises a species complex (see text); **: the two ‘sub-species’ occur in sympatry in TNP

and may actually represent two valid species; ***: an undescribed species compare Jongsma et al. (2018).

TNP habitat Agricultural areas

PF DF CP RP RF

Sample units (days) 28 12 6 4 14

Search effort (person-h) 392 168 84 56 196

Estimated species richness (Chao 2) 2.02 2 23: 3354, TSC 6 79413 100201

Estimated species richness (Jack-knife 1) 31.64£1.7 263425 16.7418 90+10 10.9409

Number of species recorded 27 23 14 7 10

Habitat Habitat

Family and species PF DF cP RP RE (adult) (tadpole)

Arthroleptidae

Arthroleptis cf. poecilonotus* x x x x Ov d

Astylosternus occidentalis xX Fl f

Cardioglossa occidentalis xX FAL f

Leptopelis occidentalis xX Ft S

Leptopelis spiritusnoctis x xX D,t S

Leptopelis viridis x O,t S

Bufonidae

Sclerophrys maculata be x x x Q,1 S

Sclerophrys regularis x x xX O,1 S

Sclerophrys togoensis x Fl f

Dicroglossidae

Hoplobatrachus occipitalis x x O,a S

Hemisotidae

Hemisus marmoratus* x x Of S

Hyperoliidae

Afrixalus dorsalis x x x. O,t S

Afrixalus nigeriensis % Et S

Hyperolius concolor x x x O,t S

Hyperolius f. fusciventris** x D,t S

Hyperolius f. lamtoensis** x Det S

Hyperolius guttulatus x xX O,t S

Hyperolius picturatus* x x B5 D,t S

Hyperolius sylvaticus x Et S

Kassina sp. x x et s?

Conrauidae

Conraua alleni x Fa ii

Pipidae

Xenopus tropicalis x D,a S

Phrynobatrachidae

Phrynobatrachus alleni ps x xX Foi S

Phrynobatrachus annulatus xX Fl ve

Phrynobatrachus calcaratus* x x O, |

Phrynobatrachus fraterculus xX xX D, 1 ?

Phrynobatrachus gutturosus* x Be O,1 S

Phrynobatrachus latifrons x x xX O, I/a S

Phrynobatrachus liberiensis xX x Fl f

Phrynobatrachus phyllophilus x Fj S

Phrynobatrachus plicatus x x FI S

Phrynobatrachus tokba xX O, | d

Amphib. Reptile Conserv. 5 January 2022 | Volume 16 | Number 1 | e298

The anuran fauna in a protected West African rainforest

Table 1 (continued). Amphibian species recorded in the Tai National Park and surrounding agricultural areas, along with recorded

and estimated species numbers per habitat type and general habitat preferences of adults and tadpoles (see Materials and Methods).

Abbreviations: PF: primary forest; DF: degraded forest; CP: mosaic of coffee and cocoa plantations; RP: rubber plantation; RF:

rice field; |: leaf litter; t: arboreal; a: aquatic; f: fossorial; F: closed forest; D: degraded forest; O: open habitat; ? : unclear as species

identification was not possible; d: direct development and non-hatching/non-feeding tadpoles; s: tadpoles in stagnant water (ponds,

puddles, tree holes); f: tadpoles in flowing water; *: comprises a species complex (see text); **: the two ‘sub-species’ occur in

sympatry in TNP and may actually represent two valid species; ***: an undescribed species compare Jongsma et al. (2018).

Family and species PF DF

Ptychadenidae

Ptychadena cf. aequiplicata*

Ptychadena bibroni

Ptychadena longirostris

Ptychadena mascareniensis*

Ptychadena pumilio

Pyxicephalidae

Aubria subsigillata x

Rhacophoridae

Chiromantis rufescens x

Ranidae

Amnirana aff. albolabris*** b4

Amnirana galamensis* x

and 17 genera (Table 1). Forests comprised more species

(27 in primary, 23 in degraded forest) than agroforestry

systems. Whereas 14 species were found in mixed

coffee/cocoa plantations and 10 species in rice fields,

only seven species were recorded in rubber plantations

(Table 1). One-third of the species only occurred in

forest (14 species, 34.1%), and 10 (24.4%) prevailed in

degraded forests; however, the majority thrived well in

open habitats (16, 39.0%). Only one species (2.4%) was

fossorial, three (9.8%) were aquatic, 12 (29.3%) were

arboreal, and the majority (23 species, 56.1%) occurred

in the leaf litter. Three species had no free-living tadpole

(4.9%), the tadpoles of seven species (17.1%) live in small

rivers, and the majority (29, 70.7%) develop in stagnant

waters of various sizes. A large proportion of frog species

were only found in primary forest (15 species, 36.6%),

but the remaining species either tolerated some habitat

degradation, or only occurred in degraded habitats (13

species, 31.7%; Table 1). The primary forest records of

two species known to usually occur in savanna habitats,

Leptopelis viridis and Ptychadena bibroni, indicates that

our “primary forest’ records also comprise ‘guests’ that do

not normally live in that habitat.

Based on the daily species lists, the overall sampling

efficiency for the entire study area was calculated (Fig.

3). The Jack-knife 1 estimator calculated 42.0 (SD: + 1.0)

amphibian species, and the Chao 2 estimator estimated

41.0 (SD: + 0.1) species for the entire study area, which

represent 97.6% and 100% of the 41 observed species,

respectively. Therefore, we would not expect to find more

species through an enhanced sampling effort. Concerning

the different habitat types, the two estimators showed

similar efficiencies 1n the respective habitats, with values

ranging from 77.8% to 100% of the actual observed

Amphib. Reptile Conserv.

Habitat Habitat

cP RE RE (adult) (tadpole)

F, 1 S

O, | S

D, | S

O, | S

Fa f

D,t Ss

D, 1 f

O, | S

species counts. The lowest value was calculated by Jack-

knife 1 for the rice fields, while the highest estimate was

calculated by Chao 2 for rubber plantations (Table 1).

The results of the Jaccard’s similarity of species

assemblages between the five surveyed habitat types

are presented in Table 2. The amphibian assemblages

in degraded forest and the coffee and cocoa plantations

were most similar with a value of 56.2%, followed by

42.9% similarity of amphibian assemblages in coffee/

cocoa with those from rubber plantations. All other

assemblages shared only a few species, e.g., the primary

and secondary forest assemblages showed only 25.0%

similarity. The lowest similarity was between primary

forest and rubber plantations (6.3%).

Discussion

The results of this study revealed higher anuran species

richness in forested habitats of Tai National Park (TNP)

compared to the surrounding agroforestry systems.

Table 2. Jaccard’s similarity of the anuran assemblages

calculated with presence/absence data in pairwise comparisons

between the five habitat types investigated in Tai National Park

and the surrounding agroforestry systems. The values show the

percentage of shared species between the two habitat types.

The highest similarity in species composition was observed

between degraded forest and coffee/cocoa plantations (in bold).

Habitat type PF DF CP RP RF

Primary forest (PF) = w2o:00872122 e625 -13:33

Degraded forest (DF) - - 56.22 30.43 20.00

Coffee/cocoa plantation (CP) _ - - - 42.85 25.00

Rubber plantation (RP) - - - - 16.67

Rice paddies (RF) - - - - -

January 2022 | Volume 16 | Number 1 | e298

Oussou et al.

Fig. 4. Selected amphibian species from Tai National Park and surrounding agroforestry systems. (A) Conraua alleni inhabits

rainforest streams. (B) Ptychadena cf. aequiplicata is a typical inhabitant of primary forest. (C) Chiromantis rufescens breeds in

puddles along forest roads and stagnant ponds and puddles in the forest. (D) Arthroleptis cf. poecilonotus (a complex of at least two

Species) occurs in primary and degraded forest, and is particularly abundant in agroforestry systems. (E) Amnirana galamensis is a

typical inhabitant of African savannas and was detected for the first time in the Tai area. (F) Ptychadena mascareniensis is a very

abundant frog in rice fields.

Whereas 27 and 24 species were detected in old growth

and degraded forests, respectively, only 14 species were

found in mixed coffee/cocoa plantations, 10 in rice fields,

and seven in rubber plantations. These observations are in

agreement with a study from Southeast Asia. In Sulawesi,

Wanger et al. (2010) observed a decrease in amphibian

species richness along a land-use gradient from closed

Amphib. Reptile Conserv.

primary forest habitats to increasingly more open areas.

In contrast, other studies revealed no significant effect

of agricultural forest exploitation on amphibian species

richness (Faruk et al. 2013; Konopik et al. 2015), with

almost identical anuran species richness 1n forested sites

and oil palm plantations in Malaysia. In fact, Ernst et

al. (2006) also observed identical amphibian species

January 2022 | Volume 16 | Number 1 | e298

The anuran fauna in a protected West African rainforest

richness in old growth forest and previously selectively

logged habitats of TNP. However, the species assemblage

composition was found to change considerably with

forest degradation (Ernst and Rodel 2005, 2006, 2008;

Hillers et al. 2008; Kpan et al. 2021), and functional

diversity was significantly lower in previously logged

forests (Ernst et al. 2006).

The impact of forest degradation on anuran

assemblages was also evident in this study, which found

the highest similarity between the degraded forest

habitats in TNP and the coffee and cocoa plantations.

Thus, the amphibians of the latter habitats clearly

occupied altered environmental conditions, which were

still ‘forest-like’ to some extent. That the presence of

shade trees and suitable breeding habitats may substitute

for forest to some degree was underscored by the

presence of some species that typically occur in proper

forest, 1.e., Phrynobatrachus alleni and Phrynobatrachus

liberiensis. However, other species recorded in those

habitats, such as Phrynobatrachus fraterculus and

Hyperolius picturatus, are typical inhabitants of

farmbush, conforming with the degradation status of

these plantations (Schigtz 1967). Rice fields provided a

home for various frog and toad species. However, all these

anurans are either typical farmbush species or species

that prefer even more open habitats, such as Ptychadena

aff. mascareniensis (compare Ndriantosa et al. 2017;

Rodel and Glos 2019). Lastly, the rubber plantations had

a very reduced amphibian fauna, with the only species

possibly reproducing there being Arthroleptis sp. and

Phrynobatrachus tokba. That these direct developing

frogs (Lamotte and Perret 1963; Barbault and Trefaut-

Rodrigues 1979; Rodel and Ernst 2002a) can benefit

from the degradation of forest has already been reported

by Ernst and Rodel (2005). With the exception of rice

fields, Arthroleptis species were found in all terrestrial

ecosystems of the TNP area. Interestingly, P. tokba was

recorded only from primary forest in this study, which

is in sharp contradiction to previous research from TNP

(e.g., Ernst et al. 2006; Kpan et al. 2021).

Based on our data, it is difficult to state whether the

species suffering from forest degradation had particular

life history traits, such as preferred micro-habitats or

larval habitats (Table 1). This is partly due to the fact that

some species were recorded exclusively in primary (e.g.,

Leptopelis viridis and Ptychadena bibroni) or degraded

forest (Amnirana galamensis) which are known to

usually occur in savanna habitats (Rodel 2000). These

records may either indicate that our plot definition was

not perfect (e.g., other neighboring habitat types with

respective species) or these specimens were ‘guests’ and

only encountered while migrating through the forested

areas. For example, savanna amphibian communities in

TNP are known to have become established on granite

inselbergs within the forest (see Figs. 1.8 and 1.33

in Rodel et al. 2021), and these species may be rarely

encountered in the forest between inselbergs (M.-O.

Amphib. Reptile Conserv.

Rodel, pers. obs. and unpub. data). However, the results

of this study clearly show that the agroforestry systems in

the TNP area could only provide habitat to a small subset

of the regional species pool, namely some generalist

forest amphibians and species that do not normally occur

in forest ecosystems. Typical members of the latter group

are species such as the three mentioned above, as well

as P. aff. mascarenisnsis and Phrynobatrachus latifrons,

which mostly occur in savanna habitats (Lamotte 1967;

Rodel 2000).

In addition to the 56 species reported by Rodel and

Ernst (2004) and Ernst et al. (2006), this study confirmed

the presence of two additional species, Conraua alleni and

Amnirana galamensis, thus raising the number of known

amphibian species for the TNP area to 58. Conraua alleni

(Fig. 4A) has been described from Liberia (Barbour and

Loveridge 1927), and reported in Céte d’ Ivoire to the south

(Rodel and Branch 2002, Haute Dodo Classified Forest)

and north of the TNP (Rodel 2003, Mont Sangbé National

Park), as well as from Mount Nimba (Guibé and Lamotte

1958; Lamotte and Perret 1968; Kanga et al. 2021). This

aquatic species prefers clear streams in forests (R6del and

Branch 2002; Rodel 2003; Rodel and Bangoura 2004).

We also observed this species in permanent streams in

primary forests with a closed canopy and open understory

in the Soubré (05°28’49”N, 07°3’35”W) and Tai sectors

(05°53’52”N, 07°23’57”W). In contrast, A. galamensis

(Fig. 4E) is generally found in savanna habitats, survives

successfully in modified habitats (Rodel 2000), and has

even been reported from urban areas (Kouamé et al. 2015).

This taxon 1s widespread in sub-Saharan A frica (Channing

and Roddel 2019), but comprises several cryptic species

(Jongsma et al. 2018). We found A. galamensis in a highly

polluted area (06°14712.7”N, 07°11704.3”W) close to a

village.

Although the estimators indicated that we could

not expect to find many more species of the local

amphibian assemblages, this seems unlikely. The

overall species richness of our study was similar to

that revealed in other nearby forest regions (see Rédel

and Branch 2002). However, our species list here

lacks a number of frogs which actually typically occur

in swampy (e.g., Phrynobatrachus villiersi, Kassina

lamottei, Hyperolius zonatus) or drier forest sites

(Phrynobatrachus guineensis), along forest streams

(Leptopelis macrotis, Hyperolius chlorosteus, Amnirana

occidentalis), and even in more open sites within forest

(Phlyctimantis boulengeri). Some of these species are

only active through particular parts of the season and/

or under certain climatic conditions, so it is possible that

the number of visits per plot was not sufficient (1.e., rice

fields) or well-suited (forested habitats) to detect them in

this study (compare Veith et al. 2004). The latter is also

indicated by the variations of our estimated sampling

efficiency. These other species which we did not detect

are generally very rare (Sclerophrys taiensis), are only

active through very particular conditions (1.e., the largest

January 2022 | Volume 16 | Number 1 | e298

Oussou et al.

rains of the season: Afrixalus vibekensis), or need rare

and hard-to-find micro-habitats (Acanthixalus sonjae in

huge water-filled tree holes; compare the publications

cited in the Introduction). However, the possibility that

these species were missed in our study plots does not

contradict our general conclusions. As almost all of the

species which we could not detect are forest species or

even forest specialists, the converse relationship between

species richness and habitat degradation might actually

be even larger than that documented in our study.

Conclusions

Our findings on the capacity (1.e., the very limited

capacity) of agroforestry systems for the maintenance of

anuran diversity has important implications for rainforest

conservation in Céte d'Ivoire and the Upper Guinea

forest zone. Our study showed that the conversion of

forest to agroforestry systems significantly reduced

species richness, in particular for the forest specialists.

However, this study also showed that some of the more

“forest-like” agroforestry systems, namely mixed coffee/

cocoa plantations with shade trees, have some capacity to

provide habitat for forest anuran communities, although

largely pauperized. Several agroforestry systems

established in the TNP area, e.g., vegetable gardens, oil

palm and banana plantations, or mixed cultivations, have

not been studied, so further research at such sites may

improve our understanding of the potential for certain

agroforestry systems in the maintenance of biodiversity.

However, our study results, in combination with the

results revealed by Ernst and Rodel (2005) and Kpan

et al. (2021), show that the amphibian assemblages in

formerly logged areas did not match those in old growth

forest even 45 years after selective logging ceased.

This clearly indicates that in order to conserve the huge

diversity of specialist and range-restricted West African

forest anurans, the remaining areas of pristine forests

need to be maintained untouched.

Acknowledgements.—We are grateful to the Ivorian

authorities, particularly the Ministere de la Salubrité, de

l’Environnement et du Développement Durable and the

Office Ivoirien des Parcs et Réserves (OIPR), for their

permission to conduct this research. We acknowledge

the Université Jean Lorougnon Guédeé for supporting this

study, and greatly acknowledge the field assistants for

their work and help. We thank Dr. Jean-Baptiste Oussou

N’Guessan for valuable comments and corrections.

Two anonymous reviewers improved our manuscript

with their constructive criticisms, and this is gratefully

acknowledged!

Literature Cited

Adou CY, Blom EC, Dengueadhé KTS, Van Rompaey

RSAR, N’Guessan EK, Wittebolle G, Bongers F.

Amphib. Reptile Conserv.

2005. Diversité Floristique et Végétation dans le

Parc National de Tai, Céte d'Ivoire. Tropenbos-Céte

dIvoire, Abidjan, Cote dIvoire. 92 p.

Adum GB, Eichhorn MP, Oduro W, Ofori-Boateng C,

Rodel M-O. 2013. Two-stage recovery of amphibian

assemblages following selective logging of tropical

forests. Conservation Biology 27: 354-363.

Alonso LE, Lauginie F, Rondeau G. (Editors). 2005.

Une évaluation biologique de deux foréts classées

du sud-ouest de la Céte d'Ivoire / A Rapid Biological

Assessment of Two Classified Forests in South-

western Céte d'Ivoire. RAP Bulletin of Biological

Assessment 34 / Bulletin RAP d’Evaluation Rapide

34. Conservation International, Washington, DC,

USA. 168 p.

Barbault R, Trefaut-Rodrigues M. 1979. Observations

sur la reproduction et la dynamique des populations

de quelques anoures tropicaux HI. Arthroleptis

poecilonotus. Tropical Ecology 20: 64—77.

Bakarr M, Bailey B, Byler D, Ham R, Olivieri S, Omland

M. 2001. From the Forest to the Sea: Biodiversity

Connections from Guinea to Togo. Conservation

Priority-setting Workshop. Conservation International,

Washington, DC, USA. 78 p.

Barbour T, Loveridge A. 1927. Some undescribed frogs

and a new gecko from Liberia. Proceedings of the

New England Zoological Club 10: 13-18.

Blackburn DC. 2010. A new Squeaker Frog

(Arthroleptidae: Arthroleptis) from Bioko Island,

Equatorial Guinea. Herpetologica 66: 320-334.

Blackburn DC, Gvozdik V, Leaché AD. 2010. A new

Squeaker Frog (Arthroleptidae: Arthroleptis) from the

mountains of Cameroon and Nigeria. Herpetologica

66: 335-348.

Bousquet B. 1978. Un parc de forét dense en Afrique:

le parc national de Tai. Revue Bois et Foréts des

Tropiques 179: 27-46.

Channing A, Rodel M-O. 2019. Field Guide to the

Frogs and other Amphibians of Africa. Struik Nature,

Penguin Random House, Cape Town, South Africa.

A08 p.

Chatelain C, Gautier L, Spichiger R. 1996. A recent

history of forest fragmentation in southwestern Ivory

Coast. Biodiversity and Conservation 5: 783-791.

Chatelain C, Kadjo B. 2000. Relations faune flore dans

le PNT: une étude bibliographique. Sempervira 9:

134-136.

Chatelain C, Kadjo B, Koné I, Refisch J. 2001. Relations

Faune-Flore dans le Parc National de Tai: une Etude

Bibliographique. Tropenbos-Cote dIvoire, Abidjan,

Cote d’Ivoire. 166 p.

Colwell RK. 2013. Statistical Estimation of Species

Richness and Shared Species from Samples. Version

9. User’s Guide and Application. Available: http://

purl.oclc.org/estimates [Accessed: 1 April 2021 ].

Dupuy B, Maitre H-F, Amsallen I. 1999. Techniques de

Gestion des Ecosystémes Forestiers Tropicaux: Etat

January 2022 | Volume 16 | Number 1 | e298

The anuran fauna in a protected West African rainforest

de l’Art. Document de Travail préparé pour la Banque

Mondiale Forest Policy Implementation Review and

Strategy. FAO, CIRAD, Rome, Italy. 146 p.

Ernst R, Rodel M-O. 2002. A new Atheris species

(Serpentes: Viperidae), from Tai National Park, Ivory

Coast. Herpetological Journal 12: 55-61.

Ernst R, Rodel M-O. 2005. Anthropogenically

induced changes of predictability in tropical anuran

assemblages. Ecology 86: 3,111-3,118.

Ernst R, Rodel M-O. 2006. Community assembly and

structure of tropical leaf-litter anurans. Ecotropica 12:

113-129.

Ernst R, R6ddel M-O. 2008. Patterns of community

composition in two tropical tree frog assemblages:

separating spatial structure and environmental effects

in disturbed and undisturbed forests. Journal of

Tropical Ecology 24: 111-120.

Ernst R, Linsenmair KE, Rodel M-O. 2006. Diversity

erosion beyond the species level: dramatic loss

of functional diversity after selective logging in

two tropical amphibian communities. Biological

Conservation 133: 143-155.

FAO. 2001. Global Forest Resources Assessment 2000:

Main Report. FAO Forestry Paper 140. FAO, Rome,

Italy. 479 p.

FAO. 2006. Global Forest Resources Assessment 2005:

Progress Towards Sustainable Forest Management.

FAO Forestry Paper 147. FAO, Rome, Italy. 320 p.

Faruk A, Belabut D, Ahmad N, Knell RJ, Garner TW.

2013. Effects of oil-palm plantations on diversity of

tropical anurans. Conservation Biology 27: 615-624.

Frost DR. 2020. Amphibian Species of the World: an

Online Reference. Version 6 (27 January 2014).

American Museum of Natural History, New

York, USA. Available: http://research.amnh.org/

herpetology/amphibia/index.html [Accessed: 23 June

2020].

Guibé J, Lamotte M. 1958. Batraciens (sauf Arthroleptis,

Phrynobatrachus et Hyperolius), in laréserve naturelle

intégrale du Mont Nimba, XII. Mémoires de I’Institut

Fondamental d’Afrique Noire Série A 53: 241-273.

Hammer 0, Harper DAT, Ryan PD. 2001. Past:

paleontological statistics software package for

education and data analysis. Palaeontologia

Electronica 4(4): 1-9.

Heyer WR, Donnelly MA, McDiarmid RW, Hayek

LAC, Foster MS. 1994. Measuring and Monitoring

Biological Diversity. Standard Methods for

Amphibians. Smithsonian Institution Press, Washing-

ton, DC, USA and London, United Kingdom. 364 p.

Hillers A, Veith M, Ro6del M-O. 2008. Effects of forest

fragmentation and habitat degradation on West

African leaf-litter frogs. Conservation Biology 22:

762-772.

Jongsma GFM, Bare) MF, Barratt CD, Burger M,

Conradie W, Ernst R, Greenbaum E, Hirschfeld M,

Leaché AD, Penner J, et al. 2018. Diversity and

Amphib. Reptile Conserv.

biogeography of frogs in the genus Amnirana (Anura:

Ranidae) across sub-Saharan Africa. Molecular

Phylogenetics and Evolution 120: 274—285.

Kanga KP, Kouamé NG, Zogbassé P, Gongomin

BA-I, Agoh KL, Kouamé AM, Konan JCBYN,

Adepo-Gourene AB, Gourene G, Rodel M-O. 2021.

Amphibian diversity of a West African biodiversity

hotspot: an assessment and commented checklist of

the batrachofauna of the Ivorian part of the Nimba

Mountains. Amphibian & Reptile Conservation 15(1)

[General Section]: 71—107 (e275).

Koné I. 2004. Effets du Braconnage sur quelques

Aspects du Comportement du Colobe bai, Procolobus

(Colobus baduis) baduis (Kerr) et du Cercopithéque

Diane, Cercopithecus diana diana (L.), dans le

Parc National de Tai, Céte-d’Ivoire. Tropenbos

International, Wageningen, Netherlands. 98 p.

Konopik O, Steffan-Dewenter I, Grafe TU. 2015. Effects

of logging and oil palm expansion on stream frog

communities on Borneo, Southeast Asia. Biotropica

47: 636-643.

Kouamé NG, Ofori-Boateng C, Adum GB, Gourene G,

Rodel M-O. 2015. The anuran fauna of a West African

urban area. Amphibian & Reptile Conservation 9(2)

[Special Section]: 1-14 (e106).

Kpan TF, Ernst R, Kouassi P, Rodel M-O. 2019.

Prevalence of endoparasitic mites on four West

African leaf-litter frogs depends on habitat humidity.

Biotropica 51: 432-442.

Kpan TF, Ernst R, Rodel M-O. 2021. Follow the forest:

slow resilience of West African rainforest frog

assemblages after selective logging. Forest Ecology

and Management 497: 119489.

Lamotte M. 1967. Les batraciens de la région de Gpakobo

(Cote d'Ivoire). Bulletin de I’Institut Francais

d’Afrique Noire Série A 29: 218-294.

Lamotte M, Perret J-L. 1963. Contribution a |’étude

des batraciens de l’Afrique de lOuest XV. Le

développement direct de lJ espece Arthroleptis

poecilonotus Peters. Bulletin de I’Institut Frangais

d’Afrique Noire Série A 25: 277-284.

Lamotte M, Perret J-L. 1968. Révision du genre Conraua

Nieden. Bulletin de I’Institut Francais d'Afrique

Noire Série A 30: 1,603—1,644.

Myers N, Mittermeier RA, Mittermeier CG, Da Fonseca

GAB, Kent J. 2000. Biodiversity hotspots for

conservation priorities. Nature 403: 853-858.

Ndriantsoa SH, Riemann JC, Raminosoa N, Ré6del M-O,

Glos JS. 2017. Amphibian diversity in the matrix

of a fragmented landscape around Ranomafana in

Madagascar depends on matrix quality. Tropical

Conservation Science 10: 1-16.

OIPR. 2006. Plan d’Aménagement et de Gestion du Parc

National de Tai. OIPR, Abidjan, Ivory Coast. 99 p.

OIPR. 2014. Plan d’Aménagement et de Gestion du Parc

National de Tai. OIPR, Abidjan, Ivory Coast. 131 p.

OIPR. 2015. Plan d’Aménagement et de Gestion du Parc

January 2022 | Volume 16 | Number 1 | e298

Oussou et al.

National de Tai. OIPR, Abidjan, Ivory Coast. 132 p.

Ofori-Boateng C, Oduro W, Hillers A, Norris K, Oppong

Sk, Adum GB, Rédel M-O. 2012. Differences in the

effects of selective logging on amphibian assemblages

in three West African forest types. Biotropica 45: 94—

101.

Perret J-L. 1988. Les espéces de Phrynobatrachus

(Anura, Ranidae) a éperon palpébral. Archives des

Sciences 41: 275-294.

Riezebos EP, Vooren AP, Guillaumet JL. 1992. Le

Parc National de Tai, Céte d’lvoire; I: Synthése

des Connaissances; II: Bibliographie. Tropenbos,

Wageningen, Netherlands. 182 p.

Rodel M-O. 1998. A new Hyperolius species from Tai

National Park, Ivory Coast (Anura: Hyperoliidae:

Hyperoliinae). Revue Francaise d’Aquariologie

Herpétologie 25: 123-130.

Rodel M-O. 2000. Herpetofauna of West Africa, Volume

I: Amphibians of the West African Savanna. Edition

Chimaira, Frankfurt am Main, Germany. 335 p.

Rodel M-O. 2003. The amphibians of Mont Sangbé

National Park, Ivory Coast. Salamandra 39: 91-110.

Rodel M-O, Bangoura MA. 2004. A conservation

assessment of amphibians in the Forét Classée du

Pic de Fon, Simandou Range, south-eastern Republic

of Guinea, with the description of a new Amnirana

species (Amphibia: Anura: Ranidae). Tropical

Zoology 17: 201-232.

Rodel M-O, Branch WR. 2002. Herpetological survey

of the Haute Dodo and Cavally forests, western Ivory

Coast, Part I: Amphibians. Salamandra 38: 245-268.

Rodel M-O, Ernst R. 2000. Bufo taiensis n. sp., eine neue

Krote aus dem Tai-Nationalpark, Elfenbeinkuste.

herpetofauna 22(125): 9-16.

Rodel M-O, Ernst R. 2001a. Description of the tadpole

of Kassina lamottei Schietz, 1967. Journal of

Herpetology 35: 678-681.

Rodel M-O, Ernst R. 2001b. Redescription of the tadpole

of Phlyctimantis boulengeri Perret, 1986, with

preliminary comments on the biology of the species.

Alytes 18: 178-186.

Rodel M-O, Ernst R. 2002a. A new reproductive mode

for the genus Phrynobatrachus: Phrynobatrachus

alticola has nonfeeding, nonhatching tadpoles.

Journal of Herpetology 36: 121-125.

Rodel M-O, Ernst R. 2002b. A new Phrynobatrachus

from the Upper Guinean rain forest, West Africa,

including a description of a new reproductive mode

for the genus. Journal of Herpetology 36: 561—571.

Roédel M-O, Ernst R. 2004. Measuring and monitoring

amphibian diversity in tropical forests. I. An

evaluation of methods with recommendations for

standardization. Ecotropica 10: 1-14.

Roédel M-O, Glos J. 2019. Herpetological surveys in

two proposed protected areas in Liberia, West Africa.

Zoosystematics and Evolution 95: 15-35.

Rodel M-O, Mahsberg D. 2000. Vorlaufige Liste der

Amphib. Reptile Conserv.

Schlangen des Tai-Nationalparks. Elfenbeinkuste und

angrenzender Gebiete. Salamandra 36: 25-38.

Rodel M-O, Schorr G, Ernst R. 2001. Zur Biologie von

Cardioglossa leucomystax (Boulenger, 1903), im

Tai-Nationalpark, Elfenbeinktste. Salamandra 37:

239-260.

Rodel M-O, Kratz D, Ernst R. 2002a. The tadpole of

Ptychadena aequiplicata (Werner, 1898) with the

description of a new reproductive mode for the genus

(Amphibia, Anura, Ranidae). A/ytes 20: 1-12.

Rodel M-O, Range F, Seppanen J-T, Noé R. 2002b.

Caviar in the rain forest: monkeys as frog spawn

predators in Tai National Park, Ivory Coast. Journal

of Tropical Ecology 18: 289-294.

Rodel M-O, Kosuch J, Veith K, Ernst R. 2003. First

record of the genus Acanthixalus Laurent, 1944 from

the Upper Guinean rain forest, West Africa, with the

description of a new species. Journal of Herpetology

37: 43-52.

Roddel M-O, Adum GB, Aruna E, Assemian NE, Bare}

MF, Bell RC, Burger M, Demare G, Doherty-

Bone T, Doumbia J, et al. 2021. Diversity, threats

and conservation of western and central African

amphibians (Senegal, The Gambia, Guinea Bissau,

Mali, Guinea, Sierra Leone, Liberia, Ivory Coast,

Burkina Faso, Ghana, Togo, Benin, Nigeria, Niger,

Cameroon, Gabon, Sao Tome and Principe, Equatorial

Guinea, Central African Republic, Chad, Republic

of the Congo, Democratic Republic of the Congo,

northern Angola). Pp. 11-101 In: Status and Threats

of Afrotropical Amphibians — Sub-Saharan Africa,

Madagascar, Western Indian Ocean Islands. Amphibian

Biology, Volume 11, Part 7 Status of Conservation and

Decline of Amphibians: Eastern Hemisphere. Frankfurt

Contributions to Natural History, Volume 78. Editors,

Heatwole H, Rodel M-O. Edition Chimaira, Frankfurt

am Main, Germany. 243 p.

Rodel M-O, Rudolf VHW, Frohschammer S, Linsenmair

KE. 2004. Life history of a West African tree-hole

breeding frog, Phrynobatrachus guineensis Guibé and

Lamotte, 1961 (Amphibia: Anura: Petropedetidae).

Pp. 31-44 In: Ecology and Evolution of Phytotelm-

Breeding Anurans. Miscellaneous Publications of the

Museum of Zoology, University of Michigan, Number

193. Editor, Lehtinen RM. University of Michigan,

Ann Arbor, Michigan, USA. 73 p.

Rudolf VHW, Rodel M-O. 2005. Oviposition site

selection in a complex and variable environment: the

role of habitat quality and conspecific cues. Oecologia

142: 316-325.

Rudolf VHW, Rodel M-O. 2007. Phenotypic plasticity

and optimal timing of metamorphosis under uncertain

time constraints. Evolutionary Ecology 21: 121-142.

Sandberger L, Feldhaar H, Lampert KP, Lamatsch DK,

Roédel M-O. 2010. Small, specialized, and highly

mobile? The tree-hole breeding frog, Phrynobatrachus

guineensis, lacks fine-scale population structure.

January 2022 | Volume 16 | Number 1 | e298

The anuran fauna in a protected West African rainforest

African Journal of Herpetology 59: 79-94. standardized transect censing. Ecotropica 10: 85-99.

Schiotz A. 1967. The treefrogs (Rhacophoridae) of | Wanger TC, Saro A, Iskandar DT, Brook BW, Sodhi NS,

West Africa. Spolia Zoologica Musei Haunienses 25: Clough Y, Tscharntke T. 2009. Conservation value

1-346. of cacao agroforestry for amphibians and reptiles

Veith M, Lotters S, Andreone F, Ré6del M-O. 2004. in South-East Asia: combining correlative models

Measuring and monitoring amphibian diversity in with follow-up field experiments. Journal of Applied

tropical forests. II. Estimating species richness from Ecology 46: 823-832.

Amphib. Reptile Conserv.

Konan Hervé Oussou is a Ph.D. student in the Department of Ecology, Biodiversity, and Evolution

at the University Jean Lorougnon Guédé (Daloa, Céte dIvoire). His current research focuses on the

diversity and ecology of anurans in the Tai National Park and its surrounding agroforestry systems in

southwestern Céte d’ Ivoire.

N’Guessan Emmanuel Assemian is a teacher-researcher (Lecturer) in the Department of Ecology,

Biodiversity, and Evolution of the University Jean Lorougnon Guédé (Daloa, Céte d’Ivoire). As an

ecologist and specialist in herpetofauna, he has been scientifically active for more than 15 years in

various studies on amphibian and reptile conservation and on the environmental impact assessments of

several development projects in diverse regions of Cote d’ Ivoire. These studies have concerned wetlands,

forests, savannahs, crops, mining sites, hydroelectric development, freshwater, and lagoon areas.

Atta Léonard Kouadio is a Ph.D. student in the Department of Ecology, Biodiversity, and Evolution

at the University Jean Lorougnon Guédé (Daloa, Céte dIvoire). His current research focuses on the

diversity and ecology of reptiles in the Tai National Park and its surrounding agroforestry systems in

southwestern Céte d’ Ivoire.

Manouhin Roland Tiédoué is a Water and Forestry Engineer who also holds a university graduate degree

in protected area management. He is in charge of ecological monitoring and the Geographic Information

System at the South-West Zone Direction of the Ivorian Office of Parks and Reserves. He implements

activities to evaluate the state of conservation of the Tai National Park and ensures collaboration with

researchers to improve our knowledge of this protected area and better guide management actions.

Mark-Oliver Rédel is the Curator of Herpetology at the Museum of Natural History in Berlin,

Germany. With his team, he is predominately focusing on the effects of changing environments, e.g., due

to logging, hunting, urbanization, climate change, etc., on species and species assemblages, mostly using

amphibians as model systems. Apart from research in Germany, he has been scientifically active for

almost 30 years in many African countries, with his research comprising such diverse fields as ecology,

conservation biology, taxonomy and systematics, biogeography, behavior, and chemical ecology.

12 January 2022 | Volume 16 | Number 1 | e298

Oussou et al.

Appendix 1. List of sampled plots with their positions in the Tai area. For ‘Sectors,’ compare Fig. 1 and GPS position in WGS 84; ‘Status’

gives the habitat status; for ‘Dates,’ all survey dates are in 2018; and ‘Habitats’ gives short habitat descriptions and the habitat type of each

plot (PF: primary forest; DF: degraded forest; CP: mosaic of coffee and cocoa plantations; RP: rubber plantation; RF: rice field).

18 May, 20 August 06°11°45.3” 07°06’07.6” Primary forest, stream eee )

19 May, 21 August 06°09°43” 07°17°59” Primary forest (drier than others; 1.e.,

no swamps) (PF)

National Park 20 May, 22 August 06°06’22” 07°19° 15” Primary forest, stream (PF)

21 May, 23 August 06°14°29” OF 13°32" Degraded forest, lake (DF)

22 May, 24 August 05°S7°30” 06°55°50” Primary forest (drier than others; 1.e.,

no swamps) (PF)

12 May, 10 August }06°11°588" 58.8” | or0e39.6" | SAG. Rice field, river, grassland (RF)

ae 13 May, 10 August 06°12°44.4” 07°03°17.1” Rice field, river, grassland (RF)

14 May, 11 August 05°57°40” 06°35" 36° Cocoa/coffee plantation (CP)

National Park 12 June, 6 October 05°28’22.0” 06°55’06.8” Primary forest (PF)

Djapadji 14 June, 9 October 05°29° 06.2" 06°53’58.6” Cocoa and rubber plantation (CP/RP)

Agricultural

system : ‘ 7 A ;

15 June, 10 } 15 June, 10 October _| 05°10’06.2 06°48’26.1 Rice field, river, grassland (RF)

20 June, 15 October 05°27°41.7” G7°13°03:3" Primary forest, stream (PF)

National Park

22 June, 16 October 05°32’ 10.0” 07°06’41.3” Primary forest, stream (PF)

Djouroutou

’ 25 June, 20 October 05°23’28.8” OT" 15203..9~ Rice field, river, grassland (RF)

Agricultural

system ) PP) 2 PP) Z *

26 June, 21 October 05°15°34.7 07°19°18.3 Rice field, river, grassland (RF)

2 July, 28 September 05°50°48.5” 06°56’33.1” Primary forest, stream (PF)

1 July, 27 September 05°44’53.0” 06°56’52.0” Primary forest, stream (PF)

30 June, 26 September } 05°38°52.0” Ge%sot4a ge _ ||| PFmare forest (drier than‘others, i.e:

no swamps) (PF)

National Park | 9 June, 25 September | 05°49°51.0” Meese sone | le at eteres. (eetnaniginers, es

no swamps) (PF)

Soubré ro} b » fe} 2 ” “

28 June, 24 September 05°53’41.0 07°06’46.0 Primary forest, stream (PF)

27 June, 23 September 05°42’37.0” 06°58’30.0” Primary forest, stream (PF)

26 June, 22 September 05°36’°04.9” OFPO02 S35” Degraded forest, stream (DF)

orioultital 28 May, 2 August 05°42’39.2 06°53714.1 Cocoa/coffee plantation (CP)

system z rs

29 May, 3 August 05°36’45.1 06°56713;1 Rubber plantation (RP)

4 June, 7 September 05°49°59.4” O7°20°32.7" Degraded forest, buildings (DF)

5 June, 9 September 05°50°04.7” 07°17°30.0” Primary forest, stream (PF)

National Park 6 June, 10 September 06°02’39.4” 07°24’46.1” Primary forest, stream (PF)

7 June, 11 September 06°03’40” 07°24’ 12” Primary forest, stream (PF)

8 June, 12 — 06°04'55.9” grcomiags || Puallary-lorseutduier than-others ae

no swamps) (PF)

21 June, 15 } 21 June, 15 September | } 0s°s428.4” 28.4” | omesasa” | 43.4” Rice field, river, grassland (RF)

eae 21 June, 28 September | 05°52°32.23” 07°27°11.26” | Rice field, river, grassland (RF)

15 June, 17 September 06°03’44.9” 07°25°46.8” Cocoa/coffee plantation (CP)

Amphib. Reptile Conserv. 13 January 2022 | Volume 16 | Number 1 | e298

Official journal website:

amphibian-reptile-conservation.org

Amphibian & Reptile Conservation

16(1) [General Section]: 14—24 (e299).

Thermal ecology of the Pygmy Alligator Lizard, Gerrhonotus

parvus Knight and Scudday, 1985 (Squamata: Anguidae), in

Nuevo Léon, Mexico

1*David Lazcano, *Javier Banda-Leal, *Héctor Gadsden-Esparza,

4Gamaliel Castaheda-Gaytan, and ‘Sandra Cecilia Hernandez-Bocardo

‘Universidad Autonoma de Nuevo Leon, Facultad de Ciencias Bioldgicas, Laboratorio de Herpetologia, Apartado Postal 157, San Nicolas de

los Garza, Nuevo Leon, C.P. 66450, MEXICO *Sistemas de Innovacion y Desarrollo Ambiental S.C. Tepeyac No. 159, Colonia Churubusco,

Monterrey, Nuevo Leon, C.P. 674590, MEXICO “Instituto de Ecologia, A.C.-Centro Regional del Bajio, Av. Lazaro Cardenas No. 253, A.P. 386,

C.P. 61600, Patzcuaro, Michoacan, MEXICO *Facultad en Ciencias Biolégicas, Universidad Judrez del Estado de Durango, Avenida Universidad

s/n, Fraccionamiento Filadelfia, C.P. 35070, Gomez Palacio, Durango, MEXICO

Abstract.—Temperature is one of the most important abiotic factors that affect organisms, and is perhaps the

most acute of all. This study investigates the thermal ecology of the Endangered lizard Gerrhonotus parvus

in Nuevo Leon, Mexico. The average body temperatures (T,) of adult males and females (24.72 + 0.79 °C and

24.10 + 1.00 °C, respectively) were not significantly different (F, ,, = 0.21, p = 0.64); and those obtained in the

spring and summer (24.50 + 0.58 °C and 25.59 + 1.38 °C, respectively) were not significantly different (F,,, =

0.66; p = 0.51). The body temperature presented positive and significant relationships with both air temperature

(T,; R? = 0.29, p < 0.05; T, = 0.55 T, + 12.52) and surface temperature (T,; R? = 0.52, p < 0.05; T, = 0.68 T, +

8.07). The slope values in the regressions of T, with air temperature (T.) and substrate temperature (T.) were

0.55 and 0.68, respectively. These results suggest that this small lizard is thigmothermic and depends more

on the temperature of the substrate (T,) than the temperature of the air (T,) to passively regulate its body

temperature. In this way, Gerrhonotus parvus obtains heat by using thermoconformity and thigmothermism,

which is consistent with the patterns presented by other species of anguids.

Keywords. Body temperature, ecophysiology, eurythermy, Reptilia, thermoconformer, thigmothermy

Citation: Lazcano D, Banda-Leal J, Gadsden-Esparza H, Castafeda-Gaytan G, Hernandez-Bocardo SC. 2022. Thermal ecology of the Pygmy

Alligator Lizard, Gerrhonotus parvus Knight and Scudday, 1985 (Squamata: Anguidae), in Nuevo Léon, Mexico. Amphibian & Reptile Conservation

16(1) [General Section]: 14-24 (e299).

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.

Accepted: 30 October 2020; Published: 20 January 2022

Introduction

Temperature is one of the most important abiotic

factors that affect organisms (Allee and Park 1939;

Sinervo et al. 2010), and is perhaps the most acute of

all (Angilletta et al. 2002; Huey et al. 2010) because it

affects all aspects of their physiological performance

(Hutchison and Dupré 1992). It can influence the

distribution and ecology of lizards since certain species

regulate their body temperature within a relatively

narrow range during their activities that corresponds to

the optimum for their metabolism, locomotion, and other

physiological functions (Angilletta et al. 2002; Bowker

and Johnson 1980). These organisms control their

body temperature by a combination of both behavioral

and physiological patterns (Bowker 1984; Hertz et al.

1982; Huey 1982). Thus, the impacts of changes in the

environmental temperature on populations depend on

the acclimatization, thermoregulatory behavior, habitat

selection, and changes in the patterns of daily activity,

in addition to changes in phenology and reproduction

(Deutsch et al. 2008; Huey and Slatkin 1976; Huey et

al. 2009; Kearney et al. 2009), and perhaps, ultimately,

by their ability to follow changes in the thermal niche

(Lara et al. 2015). Therefore, the study of thermal

ecology has become integral to our understanding of the

ecophysiology of these reptiles (Angilletta 2009; Avery

1982; Bartholomew 1982; Sinervo et al. 2010).

Thermoregulatory strategies among reptiles range

on a continuum from thermoconformity to active

thermoregulation (Huey and Slatkin 1976). Some lizard

Species are predominantly thermoconformers; so, for

example, they keep their body temperatures similar to

those of the environment (Hertz et al. 1993; Piantoni

Correspondence. *imantodes52@hotmail.com (DL), javier_banda @hotmail.com (JBL), hgadsden@gmail.com (HE), gamaliel.cg@gmail.com

(GCG), sandra. hernadez. bocardo@gmail.com (SCHB)

Amphib. Reptile Conserv.

January 2022 | Volume 16 | Number 1 | e299

Lazcano et al.

et al. 2016; Ruibal 1961; Rummery et al. 1994). Others

are accurate thermoregulators which can maintain

temperatures close to their preferred body temperature

and above the ambient temperature (Bauwens et al. 1996;

Christian 1998; Gutiérrez et al. 2010; Ibargtiengoytia et

al. 2010; Lara et al. 2015; Sartorius et al. 2002; Valdecanto

et al. 2013). Genera such as Anolis and Liolaemus

present intrageneric variation in their thermoregulatory

strategies, with some species being thermoregulators and

others thermoconformers (Piantoni et al. 2016). Thus,

observations have indicated that a spatial distribution

pattern for thermoregulation exists, such that close

to the equator and at low elevations, the incidence of

thermoconformity increases, leading to a limited capacity

for adapting to climate change (Huey et al. 2003; Sears

et al. 2011). On the other hand, Gerrhonotus species tend

to have a fragmented distribution with low abundance, so

that more information on the thermal ecophysiology of

these populations is urgently required. Such information

will allow a better understanding of the thermoregulatory

strategies that are present in various populations which

occupy different habitats and during different seasons

of the year, and an understanding of these strategies

can assist in the conservation of potentially vulnerable

populations. For this reason, the objective of the

present study is to analyze the basic thermal ecology of

Gerrhonotus parvus in northeastern Mexico.

The genus Gerrhonotus is represented in Mexico

by eight species: G. farri, G. infernalis, G. lazcanoi,

G. liocephalus, G. lugoi, G. mccoyi, G. ophiurus, and

G. parvus. Of these, the most widely distributed are G.

liocephalus in western and southern Mexico and G.

infernalis in central and northern Mexico and southern

Texas (Good 1994). The remaining species are found in

small areas and are known from only a few individuals.

Gerrhonotus ophiurus is distributed in Tamaulipas, Nuevo

Leon, central and southwestern San Luis Potosi, eastern

Querétaro, Hidalgo, Tlaxcala, Puebla, and the mountainous

areas of northern Veracruz (Lemos-Espinal and Dixon

2013; Nevarez de los Reyes et al. 2019); historical G. /ugoi

was isolated in the Basin of Cuatrociénegas, Coahuila

(McCoy 1970), but recently it has been reported in Nuevo

Leon (Garcia- Vazquez et al. 2016; Montoya-Ferrer et al.

2021); G. farri is found near Tula, Tamaulipas (Bryson

and Graham 2010); G. mccoyi is known only from the

shores of several small lagoons, and in the Basin of

Cuatrociénegas, Coahuila (Garcia-Vazquez et al. 2018);

and G. parvus is known only from four localities in Nuevo

Leon and one in Coahuila. In Nuevo Leon, it inhabits the

municipalities of Galeana, Los Rayones, Santiago, and

Santa Catarina (Banda-Leal et al. 2013, 2014b).

The four previously-mentioned small species (G. farri,

G. lazcanoi, G. lugoi, and G. parvus) have restricted

distributions and very little is known about their biology.

For G. farri and G. lazcanoi, only the collecting data

for a single specimen of each are known (Banda-Leal et

al. 2016, 2017); and for G. /ugoi, there is only a report

Amphib. Reptile Conserv.

of reproduction in captivity that describes the courtship

behavior and litter size (Lazcano et al. 1993). For G.

parvus, some details are available about its natural history

based on work that began when it was first described in

1985 (Knight and Scudday 1985; Banda-Leal et al. 2002,

2005, 2013, 2014a,b; Bryson et al. 2003; Conroy et al.

2005; Banda-Leal 2016). In a recent document, Garcia-

Vasquez et al. (2016) mentioned finding G. parvus in the

municipality of Mina, Nuevo Leon. Although efforts have

been made to understand the phylogenetic relationships of

the species in this genus (Good 1988, 1994; Conroy et al.

2005), they remain unclear. The Pygmy Alligator Lizard,

Gerrhonotus parvus, 1s an Endangered species known only

from the Sierra Madre Oriental in the states of Nuevo Leon

and Coahuila, Mexico (Fig. 1). Even though our group

has written many articles on this species during the past

decade, and much of this information was documented by

Banda-Leal (2016), much still remains to be discovered in

our understanding of the biology of G. parvus.

The characteristics of the few localities where G. parvus

has been found can provide some insights regarding its

habitat environments. The type locality of G. parvus is

in a transition zone between pine forest (Pinus arizonica)

and open gypsophyllous scrub, in a locality called Ejido

de Santa Rita. This locality is a flat portion of the ejido (a

communal piece of land), with patches of Texas Mountain

Laurel (Sophora secundiflora), dispersed individuals of St.

Peter’s Palm (Yucca filifera), and some herbaceous plants,

such as grasses and globular cacti (Coryphantha sp.,

Turbinicarpus beguinii, and Mammillaria sp.). There are

some low hillsides with steep slopes, as well as canyons

formed by streams, where limestone and chalky soils are

present. On these slopes, piedmont scrub and rosetophilous

scrub vegetation is found, and the pine community is

composed of Arizona Pine (Pinus arizonica) and Mexican

Pinyon Pine (Pinus cembroides). The elevational gradient

in this area is 1,650—1,850 m.

However, the nature of the microhabitats at the other

G. parvus localities suggest that it has a preference for dry

limestone canyons. The second locality for the species is

Cafion San Isidro, Santiago, Nuevo Leon. This canyon

lies at an elevation of 1,600-—1,750 m, runs east to west,

is characterized by steep limestone walls covered with

agaves (Agave sp.), sotols (Dasylirion sp.), and scrub oaks

(Quercus sp.), and has intermittent pools of water. The

canyon bottom has piles of leaf litter with scattered large

rocks (Banda-Leal et al. 2002; Bryson and Lazcano 2005)

where the specimens have been found. The third locality

of Cafion Mireles, Los Rayones, Nuevo Leon, consists of

piedmont scrub elements with a habitat similar to that of

Cafion San Isidro, but with an elevation of 900 m (Conroy

et al. 2005). The fourth locality is Cafion Reflexiones in the

municipality of Santa Catarina, Nuevo Leon. This narrow

canyon has an elevation of 1,650 m and is composed of

limestone rock, with the presence of rosetophilous and

piedmont scrub elements. An extensive list of the species

found here was documented by Banda-Leal et al. (2014b).

January 2022 | Volume 16 | Number 1 | e299

Thermal ecology of Gerrhonotus parvus

300000 330000

2820000

j = =f tel coment AN. 7

ae 5 ae

a AW Ale a,

canine Inte. PAN

i Wee tae rns,

Let OB fy,

ST nj a Oy, ee

ie Ip cheney AE ie ae pf. mea ry

Hf rar) Cel Slt 4 Peis

.. Coahuila

tte,

2790000

2760000

2730000

300000

330000

360000

390000 420000

360000

Gerrhonotus parvus

sv Santiago

* Galeana

|_| Current distribution

Altitude (meters)

0-300

300-500

500-800

800-1200

1200-2000

> 2000

2820000

2790000

2760000

2730000

390000 420000

Fig. 1. Distribution of Gerrhonotus parvus in northeastern Mexico. The stars indicate the localities of specimens used in this study:

Cafion de San Isidro, Santiago (white star) and Ejido Santa Rita, Galeana (black star). The coordinates are shown around the edges

of the map in the UTM/WGS§84 metric system.

The most recent findings of the species outside of

Nuevo Leon were in the municipalities of Arteaga and

Saltillo in the state of Coahuila in a Natural Protected

Area called Sierra de Zapalinamé. Here, it was found at

elevations from 1,700-—3,100 m, with the vegetation types

including desert scrub, submontane grasslands, gallery

oak, and pine forests, depending on the specific localities

where the specimens were found within the protected area.

For example, the locality of Cafion de San Lorenzo, where

most of the G. parvus were found, has one of the most

diverse floral communities in the Sierra Madre Oriental.

The dominant plant species are Sotols (Dasylirion

cedrosanum), Chaparro Oak (Quercus pringlei), Little

Bird Tree (Lindleya mespiloides), and Evergreen Sumac

(Rhus virens) growing on a rocky substrate with abundant

crevices. In the locality of the Paraje Aguajes area, the

principal vegetation 1s composed mainly of Chaparro Oak,

Mexican Drooping Juniper (Juniperus flaccid), Sotols,

Lechuguilla Agave (Agave lechuguilla), and Apak Palm

(Brahea dulcis). The other locality within the protected

area is called Cerro de las Nieves II, where the main plant

elements are Sotols, Chaparro Oak, Little Bird Tree, and

various grasses (Banda-Leal et al. 2018).

The goal of this study was to gather more information

about the thermal ecology of G. parvus in its natural

Amphib. Reptile Conserv.

habitats. Three years of field surveys were conducted

to locate specimens and obtain thermal measurements

of the lizards themselves, along with data on the air and

substrate temperatures where they were found.

Materials and Methods

Field surveys were conducted from 2012 to 2015 during

the months of March—October. The previously known

localities for G. parvus within the Sierra Madre Oriental

in Nuevo Leon were surveyed. These localities were

visited from 0800-1600 h at Ejido Santa Rita, Galeana,

4.5 km south of the entrance to the town of Galeana, in

the Cafion de Mireles, and Los Rayones, 2.20 km to the

northeast of the municipality capital of the same name,

then in Cafion de San Isidro, Santiago. Two additional

areas in which this lizard might potentially occur were

also surveyed, 1.e., Cafion de Reflexiones, Santa Catarina,

3.6 km south of the locality of Casa Blanca and the area

in the foothills of Sierra Madre Oriental, 4.73 km east of

Casa Blanca, Garcia, Nuevo Leon. A total effort of 1,400

person-hours was expended during these surveys.

The samples were assembled using the Campbell

and Christman (1982) and Dodd (2016) methods, which

consist of locating and capturing specimens on the

January 2022 | Volume 16 | Number 1 | e299

Lazcano et al.

=~!

|] Pps

hy lll lth

o800 0900 1000 1100 1200 1300 1400 1500 1600 1700 1800 1900 2000

Number of indiviuals

hm tu & tn an

T T

ANS

Time (

Fig. 2. Seasonal pattern of daily activity of Gerrhonotus parvus

in Sierra Madre Oriental.

different substrates present, either under or on rocks,

leaf litter, vegetation, and soil. The substrates were

manipulated using hybrid herpetological hooks that are

used for handling snakes (Professional Field Hook 45),

herpetological forceps (Tweezers 24), and a borescope

of 90 cm long (Extech BR200). The habitat structure

used by the species in each locality was characterized

and quantified according to the Brau-Blanquet method,

which consists of establishing the percentages of the

different structures that typify the habitat, such as rocky

areas, leaf litter, vegetation, and soil (Kent and Coker

1992; Greenberg et al. 1994; Jellinek et al. 2004).

Each specimen was captured manually or with

herpetological tweezers. Data recorded included the date,

hour, and the body temperature (T,,) during the first 10 sec

after capture, which was recorded with a thermometer

Raytek (MiniTEMP) with a sensor sensitive to + 0.1 °C

at 5 cm from the dorsal surface. The air temperature (T_)

was measured in the shade at 5 cm above the substrate

where the individual was first observed, and the substrate

temperature (T,) was measured in the shade by touching

the substrate where the individual was first seen. Also

recorded were the SVL (mm), sex, the activity of the

specimen, and location coordinates using the UTM,

WGS84 (Garmin Etrex 10) metric system.

Simple linear regression was used to analyze the

relationships of T, with T, and T_. To determine whether

this species is a thermoregulator or thermoconformer, the

criteria used by Huey and Slatkin (1976) were applied.

According to these criteria, a species is a thermoregulator

when the slope of the linear regression of the T, and the

environmental temperature (T, or T,) is zero or close

to zero, and a species is a thermoconformer when the

slope is one or close to one. In addition, if the correlation

between T, and T_ is greater than the correlation between

T, and T., then the organism is assumed to have a

heliothermic tendency; while the tendency is thought to

be thigmothermic if the opposite correlation 1s found.

Significant differences between T, and either T, or T,

were tested using an Analysis of Variance (ANOVA), and

the significance value used for all statistical tests was P <

0.05. Post-hoc pair-wise comparisons (Tukey Test) were

tested for significance. The tables and results in the text

Amphib. Reptile Conserv.

Soil

Vegetation

11%

Fig. 3. Proportions of microhabitats used by Gerrhonotus

parvus.

show the average + standard error, sample size (N), and

temperature range (Sokal and Rohlf 2000).

Results

General Observations of the Specimens Found, Their

Behaviors, and Habitats

During the course of the fieldwork, 51 active individuals

were observed at two locations: Galeana (5): two males,

two females, one unsexed; and Cafion de San Isidro (46):

28 males, 12 females, four unsexed. The most active

seasons for the species were spring (37): 22 males, 11

females, four unsexed; and summer (14): seven males,

three females, four unsexed.

The majority of the males of this species were

observed in the spring and summer, and most of the

observed specimens were adults. The activity pattern

varied seasonally. During the spring, most of the activity

was recorded between 1100-1900 h but remained

generally constant throughout the day, whereas in the

summer the activity occurred between 1000-1800 h.

Similar to other species of lizards, the pattern of activity

during the spring seems to be unimodal, whereas during

the summer it tends to be bimodal with peaks of activity

in both morning and afternoon (Fig. 2). In general, more

activity was observed after periods of rain.

Individuals were located in cool, moist, and shaded

microhabitats, mainly on rocks (65%), leaf litter (16%),

vegetation (11%), and soil (8%) (Fig. 3). In general, they

seemed to avoid direct exposure to sunlight. It is likely that

abiotic factors (such as temperature and humidity) are the

most important factors that influence the activity patterns

and microhabitat selection (Angert et al. 2002; Pal et

al. 2010). This could explain why the modeling of the

ecological niche indicated that bioclimatic isothermality

was the variable that influenced the distribution G.

parvus in the localities within the Sierra Madre Oriental.

January 2022 | Volume 16 | Number 1 | e299

Thermal ecology of Gerrhonotus parvus

Table 1. Body temperature (T,), air temperature (T,), and substrate temperature (T,) for Gerrhonotus parvus in Sierra Madre

Oriental, Nuevo Leon.

Season N

Body temperature (T,) Spring 37

Summer 14

Total 51

Air temperature (T) Spring 37

Summer 14

Total 51

Substrate temperature (T,) Spring 37

Summer 14

Total 51

These localities fall within the geographic provinces of

Gran Sierra Plegada and Sierra y Llanuras Occidentales

(Band-Leal 2016).

Temperature Measurements

Temperatures by gender. The body temperature (T,)

averages of adult males (24.72 + 0.79 °C, N = 31) and

females (24.10 + 1.00 °C, N = 15) were not significantly

different (F, ,, = 0.21, p = 0.64).

Temperatures by season. Seasonally, the body

temperatures (T,) obtained from 37 specimens in the

spring (average: 24.50 + 0.58 °C, range: 17.80—30.10

°C), and from 14 specimens in summer (average: 25.59

+ 1.38 °C, range: 13.4-32.4 °C) were not significantly

different (F, ,, = 0.66; p = 0.51, Table 1).

The air temperature (T,) averages for G. parvus sites

in the spring for 37 specimens (average: 23.59 + 0.63

°C; range: 13.80—32.00 °C) and in the summer for 14

specimens (average: 24.74 + 1.31 °C, range: 10.60—31.60

°C) were not significantly different (F, ,, = 0.84; p = 0.43,

Table 1).

The substrate temperature (T,) averages for G. parvus

sites in the spring for 37 specimens (average: 21.29 +

0.61°C; range: 15.00-—29.60 °C) and in the summer for 14

specimens (average: 23.81 + 1.18 °C; range: 11.60—30.20

°C) were not significantly different (F, ,, = 2.52; p =0.09,

Table 1).

2,49

Correlations between the temperatures. The ANOVA

results showed differences between T,, T., and T CE. ‘in

11.27; p = 0.004). Pair-wise comparisons (Tukey Test)

showed that the average body temperature was different

than the average substrate temperature (q = 4.544, p <

0.05), the average body temperature was not different

than the average air temperature (¢ = 1.08, p > 0.05),

and the average air temperature was different than the

average substrate temperature (g = 3.46, p < 0.05).

The body temperature presented a positive and

significant relationship with T, (R* = 0.29, p < 0.05;

T, = 0.55T, + 12.52) and with T, (R? = 0.52, p < 0.05;

T, = 0.68T, + 8.07) (see Fig. 4). The slope value for

Amphib. Reptile Conserv.

Mean SE Minimum Maximum

24.50 0.58 17.80 30.10

25.59 1.38 13.40 32.40

24.80 0.56 13.40 32.40

23.59 0.63 13.80 32.00

24.77 1.31 10.60 31.60

23.92 0.58 10.60 32.00

21.29 0.61 15.00 29.60

23.81 1.18 11.60 30.20

21.91 0.56 11.60 30.20

the regression of the body temperature (T,) and air

temperature (T_,) was 0.55, and for the regression of the

body temperature (T,) and substrate temperature (T,) it

was (0.68.

Discussion

The body temperatures recorded for active males

and females of G. parvus (24.72 + 0.79 °C and 24.10

+ 1.00 °C, respectively) are within the range of the

optimum thermal gradient for individuals of the genus

Gerrhonotus, which ranges from 21 to 32 °C (http://

madisonherps.org/guwp/wp-content/uploads/2016/07/

AlligatorLizards. pdf).

The lack of a significant difference between the body

temperatures (T,) of females and males of G. parvus (Fig.

5) was also observed by Fierro-Estrada (2013) for Abronia

taeniata. This feature may be due to the fact that the two

sexes occupy very similar microhabitats. In another

species of the same genus (Gerrhonotus infernalis),

Garcia-Bastida (2013) observed male-female couples

sharing the same place of refuge for several weeks, which

might be the same situation for G. parvus. On the other

hand, G. infernalis has an average body temperature of

22.9 °C in spring and summer (Garcia-Bastida 2013).

This species tends to occur in more shaded microhabits

o y=0.55x + 12.52

R? = 0.29 A °

- T, vs 1, : 0,

A y=0.68x + 8.07 a7¢°

R?=0.52 & y = On 4

T, (°C)

8 13 18

28 33

T, and T, (°C)

Fig. 4. Relationship between body temperature (T,), air

temperature (T_) and substrate temperature (T,) for Gerrhonotus

parvus of Sierra Madre Oriental in Nuevo Leon, Mexico.

January 2022 | Volume 16 | Number 1 | e299

Lazcano et al.

Fig. 5. Specimens of (A) male Gerrhonotus parvus and (B) female Gerrhonotus parvus.

Table 2. Average body temperature (T,) and relationships of body temperature with substrate temperature (T,) and air temperature

(T,) in several species of Anguidae. An asterisk (*) indicates body temperature was taken in the shade.

Species

Barisia imbricata

Abronia taeniata

Gerrhonotus infernalis

Elgaria paucicarinata

Gerrhonotus parvus

Average

T,CO)

26.6

22.4

22:3

20.7

25.4

25.0

Amphib. Reptile Conserv.

Range T,

12-34

11-30

14-30

18-34

20-31

13-32

R’ of

T, VS. T,

0.43

0.18

0.73

0.98

0.84

0.52

R’ of

¥ VS. T,

0.30

0.20

0.76

0.83

0.80

0.29

References Thermoregulatory trends

Lemos-Espinal et al. 1998

Murioz-Brito 2013 Thigmothermic, eurythermic

Fierro-Estrada Thermoconforming

2013 Facultative eurythermic

Garcia-Bastida Thermoconforming

2013 Thigmothermic, eurythermic

Valdez-Villavicencio and Galina- Thermoconforming

Tessaro 2014 Thigmothermic

This study Thermoconforming

Thigmothermic, eurythermic

January 2022 | Volume 16 | Number 1 | e299

Thermal ecology of Gerrhonotus parvus

than G. parvus to avoid direct contact with the sun’s

rays, and it occupies much higher rocky microhabitats

that are more exposed to the sun when it 1s active.

Differences in the average body temperatures measured

for these two species are perhaps due to the differences

in the weather conditions and the elevations of the areas

where they occur. Gerrhonotus parvus inhabits a variety

of plant communities in arid mountainous areas, such as

pine-oak forest, pine forest, and oak forest in transition

(http://www.fcb.uanl.mx/herpetologia); and is more

commonly found at 1,600—1,650 m (Canseco-Marquez

and Mendoza-Quijano 2007) and especially among

rocks and within leaf litter. In contrast, G. infernalis is

most abundant at elevations of 1,360—3,400 m, usually

occupying rock crevice microhabitats, and 1s distributed

from semi-desert regions to rocky pine forests (Good

1988; Lemos-Espinal et al. 2018). As in G. infernalis,

there is an increase in the activity of G. parvus in the

spring and summer months after the rains.

With respect to its trends as a thermoregulator, two

pieces of evidence might indicate a passive temperature

thermoconformism in G. parvus. The first is that the slope

of the regression for T, vs. T, (0.55) was less than the

slope for T, vs. T, (0.68). Based on the criteria proposed

by Huey and Slatkin (1976), this species presents a trend

toward the passive thermoregulator strategy (Garcia-Rico

et al. 2015). Being a thermoconforming lizard has certain

implications. For example, when not exposed to the

sun, it becomes less conspicuous to potential predators

(Huey and Slatkin 1976). On the other hand, having

lower body temperatures than other lizards, as do many

of the members of the genus Sceloporus, gives them the

benefit of losing less water to evaporation (Hertz 1992).

We can also assume that the trend of G. parvus and other

species of anguids (Barisia imbricata, Abronia taeniata,

Gerrhonotus_ infernalis, and Elgaria paucicarinata)

to show activity at low temperatures (Table 2) is an

adaptive strategy that allows them to make the best use

of the resources in the habitat, with less competition for

food and space.

The second piece of evidence that might indicate

passive thermoregulation, according to the criteria of

Huey and Slatkin (1976), is that this species has a low

correlation for T, vs. T, (R? = 0.29), which is lower than

that for T, vs. T, (R* = 0.52). This pattern suggests that

G. parvus is thigmothermic and depends more on the

temperature of the substrate (T,) than the temperature of

the air (T_) to passively regulate its body temperature (T,).

In this way, it obtains its heat by using thermoconformity

and a thigmothermic process, which is consistent with

the patterns presented by other species of anguids, such

as Abronia taeniata (Fierro-Estrada 2013), Elgaria

paucicarinata (Valdez-Villavicencio and Galina-Tessaro

2014), and Gerrhonotus infernalis (Garcia-Bastida

2013). Likewise, the members of the related family

Xenosauridae and its single genus Xenosaurus also tend

to be thermoconforming and thigmothermic species due

Amphib. Reptile Conserv.

to their extremely secretive habits (Woolrich-Pifa et al.

2012).

Other authors have found relationships between

ambient temperatures and the body temperature in other

species of the family Anguidae, such as Gerrhonotus

multicarinatus (Cunningham 1966) and Mesaspis

monticola (Vial 1975). Lemos-Espinal et al. (1998),

however, found no _ correlation between ambient

temperatures and body temperatures of Barisia imbricata.

We must highlight that in this study we found active

specimens of G. parvus at 13.4 °C, which coincides

with data recorded by Fierro-Estrada (2013) for Abronia

taeniata and those published for other species in family

Anguidae. According to Fierro-Estrada (2013), activity

at such low temperatures in the family Anguidae

suggests that these species possess certain physiological

characteristics which allow them to be active at

temperatures below the average T, of many lizards.

Conclusions

Gerrhonotus parvus and the majority of anguid species

that are thermoconformers tend to change their body

temperature as the environmental temperature of their

refuge changes, or with some degree of exposure to the

environment. Likewise, this species and the majority of

members of the family Anguidae are thigmothermic,

and can passively capture heat by conduction from the

surface with which they are in contact. Finally, we must

consider that the wide range of body temperatures of this

lizard and many other anguids gives them the possibility

of exhibiting eurythermy, so they will probably respond

adequately to the consequences of climate change. In fact,

predictive models have determined that the Anguidae

family is least vulnerable to the effects of global warming

(Sinervo et al. 2010).

Acknowledgments.—We would like to thank the San

Antonio Zoological Gardens and Aquarium, Los Angeles

Zoo and Botanical Gardens, Bioclon S.A. de C.V., and

the Universidad Autonoma de Nuevo Leon, Facultad de

Ciencias Biologicas, for financial support for this study;

and SEMARNAT for issuing collecting permits and

providing the most recent permits (Oficio Num.SGPA/

DGVS/0511/12 and 01589/13). We also would like to

thank all the persons who participated in the laboratory

and fieldwork.

Literature Cited

Allee W, Park C. 1939. Concerning ecological principles.

Science 89: 166-169.

Angert AL, Hutchinson D, Glossip D, Losos JB. 2002.

Microhabitat use and thermal biology of the collared

lizard (Crotaphytus collaris collaris) and the fence

lizard (Sceloporus undulatus hyacinthinus) in Missouri

Glades. Journal of Herpetology 36: 23-29.

January 2022 | Volume 16 | Number 1 | e299

Lazcano et al.

Angilletta Jr MJ. 2009. Thermal Adaptation: a Theoretical

and Empirical Synthesis. Oxford University Press,

New York, New York, USA. 289 p.

Angilletta Jr MJ, Niewiarowski PH, Navas CA. 2002. The

evolution of thermal physiology in ectotherms. Journal

of Thermal Biology 27. 249-268.

Avery RA. 1982. Field studies of body temperatures

and thermoregulation. Pp. 93-166 In: Biology of the

Reptilia, Volume 12. Editors, Gans C, Pough FH.

Academic Press, New York, New York, USA. 536 p.

Banda-Leal J. 2016. Taxonomia y ecologia de la Lagartija

Cocodrilo Pigmea Gerrhonotus parvus (Knight and

Scudday, 1985). Ph.D. Dissertation, Universidad

Autonoma de Nuevo Leon, Facultad de Ciencias

Biologicas, San Nicolas de los Garza, Nuevo Leon,

Mexico. 121 p.

Banda-Leal J, Bryson Jr RW, Lazcano D. 2002. New

record of Elgaria parva (Lacertilia: Anguidae) from

Nuevo Leon, Mexico. The Southwestern Naturalist 47:

614-615.

Banda-Leal J, Bryson Jr RW, Lazcano D. 2005.

Gerrhonotus parvus (Pygmy Alligator Lizard).

Maximum size. Herpetological Review 36: 449.

Banda-Leal J, Lazcano D, Nevarez-de los Reyes M. 2013.

Notes on Mexican herpetofauna 19: herpetofauna

sympatric with Gerrhonotus parvus in San Isidro

Canyon, Santiago, Nuevo Leon, Mexico. Bulletin of the

Chicago Herpetological Society 48: 13-19.

Banda-Leal J, Lazcano D, Nevarez-de los Reyes

M, Barriga-Vallejo C. 2014a. Notes on Mexican

herpetofauna 20: potential herpetofaunal predators

of Gerrhonotus parvus in the San Isidro Canyon,

Santiago, Nuevo Leon, Mexico. Bulletin of the Chicago

Herpetological Society 49: 17-23.

Banda-Leal J, Lazcano D, Nevarez-de los Reyes M,

Barriga- Vallejo C. 2014b. Gerrhonotus parvus (Knight

and Scudday, 1985, Squamata: Anguidae): new range

extension and clutch size in the state of Nuevo Leon,

Mexico. Check List 10(3): 950-953.

Banda-Leal J, Nevarez-de los Reyes M, Bryson R Jr. 2017.

A new species of Pigmy Alligator Lizard (Squamata:

Anguidae) from Nuevo Leon, Mexico. Journal of

Herpetology 51: 223-226.

Banda-Leal J, Lazcano D, Barriga-Vallejo C, Nevarez-de

los Reyes M. 2018. New records of Gerrhonotus parvus

Knight and Scudday, 1985 (Squamata, Anguidae) in

the state of Coahuila, Mexico. Check List 14: 523-528.

Bartholomew GA. 1982. Physiological control of

temperature. Pp.167—211 In: Biology of the Reptilia,

Volume 12. Editors, Gans C, Pough FH. Academic

Press, New York, New York, USA. 536 p.

Bauwens D, Hertz PE, Castilla AM. 1996. Thermo-

regulation in a lacertid lizard: the relative contributions

of distinct behavioral mechanisms. Ecology 77: 1,818—

1,830.

Bowker RG. 1984. Precision of thermoregulation of some

African lizards. Physiological Zoology 57. 401-412.

Amphib. Reptile Conserv.

Bowker RG, Johnson OW. 1980. Thermoregulatory

precision in three species of Whiptail Lizards

(Lacertilia: Teiidae). Physiological Zoology 53: 176—

185.

Bryson RW, Graham MR. 2010. A new Alligator Lizard

from northeastern Mexico. Herpetologica 66: 92-98.

Bryson RW, Lazcano D. 2005. The Pygmy Alligator

Lizard of Nuevo Leon, Mexico. Reptilia-The European

Herpetological Magazine 39: 69-72.

Bryson RW, Lazcano D, Banda-Leal J, Castafieda-Gaitan

G, Garcia-de la Pefia C. 2003. Historia natural de la

Lagartija Pigmea (E/garia parva) endémica de Nuevo

Leon, México. Boletin de la Sociedad Herpetoldgica

Mexicana 11: 21-22.

Campbell HW, Christman SP. 1982. Field techniques

for herpetofaunal community analysis. Pp. 193-200

In: Herpetological Communities. Wildlife Research

Report 13. Editor, Scott NJ. United States Fish and

Wildlife Service, Washington, DC, USA. 239 p.

Canseco-Marquez L, Mendoza-Quiyano F. 2007.

Gerrhonotus parvus. The IUCN Red List of Threatened

Species 2007: T63711A 12708470.

Christian KA. 1998. Thermoregulation by the Short-

horned Lizard (Phrynosoma douglassi) at high

elevation. Journal of Thermal Biology 23: 395-399.

Conroy CJ, Bryson R Jr W, Lazcano D, Knight A. 2005.

Phylogenetic placement of the Pygmy Alligator Lizard

based on mitochondrial DNA. Journal of Herpetology

39: 142-147.

Cunningham JD. 1966. Thermal relations of the

Alligator Lizard Gerrhonotus multicarinatus webbi.

Herpetologica 22: \—7.

Deutsch CA, Tewksbury JJ, Huey RB, Sheldon KS,

Ghalambor CK, Haak DC, Martin PR. 2008. Impacts

of climate warming on terrestrial ectotherms across

latitudes. Proceedings of the National Academy of

Sciences of the United States of America 105: 6,668—

6,672.

Dodd CK Jr. 2016. Reptile Ecology and Conservation.

A Handbook of Techniques. Oxford University Press,

Oxford, United Kingdom. 462 p.

Fierro-Estrada N. 2013. Ecologia térmica de Abronia

taeniata (Reptilia: Anguidae) y su susceptibilidad ante

el calentamiento global. Masters Thesis, Universidad

Nacional Autonoma de México, Mexico, DF, Mexico.

Garcia-Bastida M. 2013. Aspectos ecologicos de

Gerrhonotus infernalis (Sauria: Anguidae) en el

“Parque Ecolégico Chipinque,’ San Pedro Garza

Garcia, Nuevo Leon, México. Doctoral Dissertation,

Universidad Autonoma de Nuevo Leon, Facultad de

Ciencias Bioldgicas, San Nicolas de los Garza, Nuevo

Leon, Mexico.

Garcia-Rico J, Diaz de la Vega-Pérez AH, Smith GR,

Lemos-Espinal JA, Woolrich-Pifa GA. 2015. Thermal

ecology, sexual dimorphism, and diet of Xenosaurus

tzacualtipantecus from Hidalgo, Mexico. Western

North American Naturalist 75(2). 209-217.

January 2022 | Volume 16 | Number 1 | e299

Thermal ecology of Gerrhonotus parvus

Garcia-Vazquez UO, Contreras-Arquieta A, Trujano-

Ortega M, Nieto-Montes de Oca A. 2018. A new species

of Gerrhonotus (Squamata: Anguidae) from the Cuatro

Ciénegas Basin, Coahuila, Mexico. Herpetologica 74:

269-278.

Garcia-Vazquez UO, Garcia-Padilla E, Herrera-Enriquez

GJ. 2016. First record of the Alligator Lizard

Gerrhonotus lugoi (Squamata: Anguidae) for the state

of Nuevo Leon, Mexico. Primer registro de la lagartija

Gerrhonotus lugoi (Squamata: Anguidae) para el

estado de Nuevo Leon, México. Revista Mexicana de

Biodiversidad 87. 1,399-1,401.

Good DA. 1988. Phylogenetic relationships among

gerrhonotine lizards, an analysis of external

morphology. University of California Publications in

Zoology 121: 1-139.

Good DA. 1994. Species limits in the genus Gerrhonotus

(Squamata: Anguidae). Herpetological Monographs 8:

180-202.

Greenberg CH, Neary DG, Harris LD. 1994. Effect of high-

intensity wildlife and silvicultural treatments on the

reptile community in a sand-pine scrub. Conservation

Biology 8: 1,047—1,075.

Gutierrez JA, Krenz JD, Ibargtiengoytia NR. 2010. Effect

of altitude on thermal responses of Liolaemus pictus

argentinus in Argentina. Journal of Thermal Biology

35: 332-337.

Hertz PE. 1992. Evaluating thermal resource partitioning

by sympatric lizards Anolis cooki and A. cristatellus:

a field-test using null hypotheses. Oecologia 90: 127—

136.

Hertz PE, Huey RB, Nevo E. 1982. Fight versus flight-

body temperature influences defensive responses of

lizards. Animal Behaviour 30: 676-679.

Hertz PE, Huey RB, Stevenson RD. 1993. Evaluating

temperature regulation by field-active ectotherms: the

fallacy of the inappropriate question. 7he American

Naturalist 142: 796-818.

Huey RB. 1982. Temperature, physiology, and ecology of

reptiles. Pp. 25-91 In: Biology of the Reptilia, Volume

12. Editors, Gans C, Pough FH. Academic Press, New

York, New York, USA. 536 p.

Huey RB, Slatkin M. 1976. Costs and benefits of lizard

thermoregulation. The Quarterly Review of Biology 51:

363-384.

Huey RB, Hertz PE, Sinervo B. 2003. Behavioral drive

versus behavioral tnertia in evolution: a null model

approach. The American Naturalist 161: 357-366.

Huey RB, Deutsch CA, Tewksbury JJ, Vitt LJ, Hertz PE,

Perez HJA, Garland T. 2009. Why are tropical forest

lizards vulnerable to climate warming? Proceedings

of the Royal Society of London B: Biological Sciences

276: 1,939-1,948.

Huey RB, Losos JB, Moritz C. 2010. Are lizards toast?

Science 328: 832-833.

Hutchinson VH, Dupré RK. 1992. Thermoregulation.

Pp. 206-249 In: Environmental Physiology of the

Amphib. Reptile Conserv.

Amphibians. Editors, Feder ME, Burggren WW.

University of Chicago Press, Chicago, Illinois, USA.

646 p.

Ibarguengoytia NR, Medina M, Fernandez JB, Gutiérrez

JA, Tappari F, Scolaro A. 2010. Thermal biology of

the southernmost lizards in the world: Liolaemus

sarmientoi and Liolaemus magellanicus from

Patagonia, Argentina. Journal of Thermal Biology 35:

21-27.

Jellinek S, Driscoll DA, Kirkpatrick JB. 2004.

Environmental and vegetation variables have a

greater influence than fragmentation in structuring

in structuring lizard communities in remnant urban

bushland. Austral Ecology 29: 294-304.

Kearney M, Shine R, Porter WP. 2009. The potential for

behavioral thermoregulation to buffer “cold-blooded”

animals against climate warming. Proceedings of the

National Academy of Sciences of the United States of

America 106: 3,835—3,840.

Kent M, Coker P. 1992. Vegetation Description and

Analysis. A Practical Approach. Belhaven Press,

London, United Kingdom. 414 p.

Knight RA, Scudday JF. 1985 A new Gerrhonotus

(Lacertilia: Anguidae) from the Sierra Madre Oriental,

Nuevo Leon, Mexico. The Southwestern Naturalist 30:

89-94.

Lara-Reséndiz RA, Larrain-Barrios BC, Diaz de la Vega-

Pérez AH, Meéndez-de la Cruz FR. 2014. Calidad

térmica a través de un gradiente altitudinal para una

comunidad de lagartijas en la sierra del Ajusco y el

Pedregal de San Angel, México. Revista Mexicana de

Biodiversidad 85: 885-897.

Lara-Reséndiz RA, Gadsden H, Rosen PC, Sinervo B,

Méndez-de la Cruz FR. 2015. Thermoregulation of two

sympatric species of horned lizards in the Chihuahuan

Desert and their local extinction risk. Journal of

Thermal Biology 48: 1-10.

Lazcano D, Contreras-Arquieta A, Nevarez de los Reyes M.

1993. Notes on Mexican herpetofauna 3: reproductive

biology of Gerrhonotus lugoi, an anguid lizard from

the Cuatro-Cienegas Basin, Coahuila, Mexico. Bulletin

Chicago Herpetological Society 28: 263-265.

Lemos-Espinal JA, Dixon JR. 2013. Amphibians and

Reptiles of San Luis Potosi. Eagle Mountain Publishing,

Eagle Mountain, Utah, USA. 300 p.

Lemos-Espinal JA, Smith GR, Ballinger RE. 1998.

Temperature relationships of the lizard, Barisia

imbricata, from Mexico. Amphibia-Reptilia 19: 95-99.

McCoy CJ. 1970 A new Alligator Lizard (genus

Gerrhonotus) from the Cuatro-Cienegas Basin,

Coahuila, Mexico. Zhe Southwestern Naturalist 15:

37-44.

Montoya-Ferrer D, Lazcano D, Banda-Leal J, de Luna-

Gonzalez JM, Lopez-Villa PE, Wilson LD. 2021. Notes

on the herpetofauna of Mexico 37: range extension for

Lugo’s Alligator Lizard (Gerrhonotus lugoi, McCoy,

1970; Squamata: Anguidae), with commentary on its

January 2022 | Volume 16 | Number 1 | e299

Lazcano et al.

ecological traits. Bulletin of the Chicago Herpetological

Society 56(2): 17-20.

Mufioz-Brito A. 2013. Termorregulacion en Barisia

imbricata (Sauria: Anguidae). Masters Thesis, Instituto

de Biologia, Universidad Nacional Autonoma de

México, Mexico, DF, Mexico.

Nevarez de los Reyes M, Lazcano D, Wilson LD. 2019.

Gerrhonotus ophiurus (Smooth-headed Alligator

Lizard): microhabitat. Herpetological Review 50(3):

SH:

NORMA Oficial Mexicana. 2010. Proteccién Ambiental-

especies Nativas de México de Flora y Fauna

Silvestres-Categorias de Riesgo y Especificaciones

para su Inclusion, Exclusién o Cambio-Lista de Species

en Riesgo. NOM-059-SEMARNAT-2010. Diario Oficial

de la Federacion, Jueves 30 de Diciembre de 2010.

Gobierno de México, Ciudad de México, Mexico.

Pal A, Swain MM, Rath S. 2010. Observations on

microhabitat use and activity patterns in Sitania

ponticeriana (Sauria: Agamidae). Russian Journal of

Herpetology 17(1): 22-30.

Piantoni C, Navas CA, Ibargtiengoytia NR. 2016.

Vulnerability to climate warming of four genera of

New World iguanians based on their thermal ecology.

Animal Conservation 19: 391-400.

Ruibal R. 1961. Thermal relations of five species of

tropical lizards. Evolution 15: 98-111.

Rummery C, Shine R, Houston DL, Thompson MB. 1994.

Thermal biology of the Australian Forest Dragon,

Hypsilurus spinipes (Agamidae). Copeia 1994: 818—

827.

Sartorius SS, Do Amaral JPS, Durtsche RD, Deen CM,

Lutterschmidt WI. 2002. Thermoregulatory accuracy,

precision, and effectiveness in two sand-dwelling

lizards under mild environmental conditions. Canadian

Journal of Zoology 80: 1,966-1,976.

Sears MW, Raskin E, Angilletta MJ. 2011. The world 1s

not flat: defining relevant thermal landscapes in the

context of climate change. /ntegrative and Comparative

Biology 51: 662-665.

Sinervo B, Méndez-de-la-Cruz F, Miles DB, Heulin B,

Bastiaans E, Villagran-Santa Cruz M, Lara-Reséndiz

R, Martinez-Méndez N, Calderon-Espinosa ML, Meza-

Lazaro RN, et al. 2010. Erosion of lizard diversity by

climate change and altered thermal niches. Science

328: 894-899.

Sokal RR, Rohlf FS. 2000. Biometry. Freeman and

Company, New York, New York, USA. 887 p.

Valdecanto S, Martinez V, Lobo F, Cruz FB. 2013.

Thermal biology of Liolaemus lizards from the high

Andes: being efficient despite adversity. Journal of

Thermal Biology 38: 126-134.

Valdez-Villavicencio JH, Galina-Tessaro P. 2014. Elgaria

paucicarinata: field and preferred body temperatures.

Herpetological Review 45: 495.

Vial JL. 1975. Thermal related activity

Mesoamerican Gerrhonotus monticolus.

Journal of Herpetology 55: 491-495.

in the

British

Amphib. Reptile Conserv.

David Lazcano is a Herpetologist with Bachelor’s degrees in Chemical Science (1980) and Biology

(1982), a Master’s degree in Wildlife Management (1999), and a Ph.D. degree in Biological Sciences

with a specialty in Wildlife Management (2005), all from the Facultad de Ciencias Bidlogicas,

Universidad Autonoma de Nuevo Leon (FCB/UANL), Mexico. Now a full-time professor at UANL,

he has taught courses in soil sciences, herpetology, ecology, animal behavior, biogeography, the biology

and diversity of chordates, and wildlife management, and he has been teaching and providing assistance

in both undergraduate and graduate programs since 1979. David has been Head of the Laboratorio de

Herpetologia since 1993 and Coordinacion de Intercambio Académico de la Facultad de Ciencias

Bioldgicas at UANL. He received the Joseph Lazlo Award from the HIS in 2006 for his herpetological

career, and was awarded national recognition by the Asociacion para la Investigacion y Conservacion

de Anfibios y Reptiles (AICAR) in 2017 for his contributions in the ecology and conservation of

herpetofauna in northeastern Mexico (Tamaulipas, Nuevo Leon, and Coahuila). He participated in

developing the Program of Action for the Conservation of the Species (PACE) Rattlesnakes (Crotalus

sp.). His research interests include the herpetofaunal diversity of northeastern Mexico, as well as

the ecology, herpetology, biology, biogeography, behavior, and population maintenance techniques

of montane herpetofauna. He has advised many Bachelor’s, Master’s, and Ph.D. degree students on

projects dealing with the regional and national herpetofauna. David has published more 250 scientific

notes and articles concerning the herpetofauna of the northeastern portion of Mexico in indexed and

general diffusion journals. His students named the species Gerrhonotus lazcanoi in honor of his work.

Javier Banda-Leal is a Biologist who obtained a Ph.D. with an emphasis in Wildlife Management

and Sustainable Development from the Facultad de Ciencias Bioldgicas, Universidad Autonoma de

Nuevo Leon, Mexico (FCB/UANL). Javier has carried out various activities related to wildlife, mainly

in herpetology. He was Curator of the Herpetological Collections of the VANL, a founding member of

the student chapter of wildlife managers (AMAVISI), northern member of the Mexican Herpetological

Society (SHM), as well as a Founding Member and research member of the Coahuilense Association

of Speleology A.C. In connection with the latter, he has explored different underground systems in

Coahuila and other parts of Mexico. He has participated in research, management, and conservation

of reptiles; worked in the Directorate of State Parks and Natural Resources, Directorate of Protected

Natural Areas Metropolitan Area of Monterrey; developed and managed the Herpetario of the Museo

23 January 2022 | Volume 16 | Number 1 | e299

Amphib. Reptile Conserv.

Thermal ecology of Gerrhonotus parvus

del Desierto in Saltillo, Coahuila; was technical manager of the Parque La Casa de los Loros of

Monterrey and coordinator of strategic planning and scientific development in the Directorate of

Tamaulipean Ecoregion and Wetlands, in Pronatura Noreste A.C. Javier has published articles in many

national and international scientific journals, including the description of a new species of Crocodile

Lizard of the genus Gerrhonotus from Nuevo Leon. He is co-author of the book Serpientes de Nuevo

Leon. His projects have involved documenting the herpetofauna Tamaulipas, Nuevo Leon, and

various parks and natural areas; the eradication of exotic fauna and vegetation in the Valley of Cuatro

Ciénegas, Coahuila; searching and documenting the Flat-headed Bat (Myotis planiceps) in Nuevo

Leon, Coahuila, and Zacatecas; and establishing a colony of Mountain Bells for studies in captivity

and the extraction of its toxin. He has been an environmental consultant in various energy projects

that involve the analysis of populations, and the rescue and relocation of wildlife, and currently works

as an environmental consultant at the company Sistemas de Innovacion y Desarrollo Ambiental S.C.

Héctor Gadsden Esparza is a retired Senior Researcher at Instituto de Ecologia, A.C., where he

worked for 32 years, and had been a member of Sistema Nacional de Investigadores (1996-2019).

He completed his Bachelor’s, Master’s, and Ph.D. at the Facultad de Ciencias-Universidad Autondma

Nacional de Mexico (UNAM), finishing in 1988; a Master’s degree in Filosofia de las Ciencias at the

Universidad Autonoma Metropolitana (UAM), finishing in 1987; and a post-doctoral appointment at

the Instituto de Biologia-7 UNAM (1996-1997). He taught on the subjects of Evolution, Population

Genetics, Population Ecology and Taxonomy at the Facultad de Ciencias-UNAM, UAM, and

INECOL,; and was Director of “La Michilia” Biosphere Reserve in Durango (1989-1991) and Director

of INECOL-Centro Regional Chihuahua (1999-2003). Héctor has published 140 papers and scientific

notes, three books, and 14 book chapters. He has coordinated various projects financed by CONACYT

and CONABIO, and was thesis advisor for 54 Bachelor’s, Master’s, and Ph.D. students. His research

has focused on the ecology of reptile populations and assemblages in arid northern México, and the

effect of global climate change on them. Due to his outstanding career and contributions to Mexican

herpetology, Héctor was awarded national recognition by the Asociacion para la Investigacion y

Conservacion de Anfibios y Reptiles (AICAR) in October 2017. For his contributions in the ecology

and conservation of Chihuahuan Desert herpetofauna, a new species of lizard (Sce/oporus gadsdeni)

was dedicated to him in 2017. Finally, in November 2019, the Universidad Juarez del Estado de

Durango (UJED) dedicated its 3“ Congress on Biological Diversity to him, where he received this

special recognition.

José Gamaliel Castaiieda Gaytan is a Biologist from the Escuela de Biologia, Universidad Juarez del

Estado de Durango, México (UJED). He obtained his Ph.D. from the Facultad de Ciencias Bioldgicas,

Universidad Autonoma de Nuevo Leon (FCB/UANL), Mexico, in 2007. He has participated in more

than 30 national and international congresses on various topics, and has authored or co-authored more

than 50 scientific refereed articles published in national and international journals, in addition to other

notes on geographical distribution and natural history and collaborations in books and conference

proceedings. José participated in developing the Program of Action for the Conservation of the Species

(PACE) Rattlesnakes (Crotalus sp.), and has collaborated on seven multidisciplinary research projects

focusing on biodiversity and conservation in protected natural areas of the region. He is a member of

the North American Box Committee for Turtle Conservation, and has been a referee on articles in many

different scientific journals. José was Associate Editor of the Bulletin of the Mexican Herpetological

Society (2008-2009), then Editor-in-Chief of the Bulletin of the Mexican Herpetological Society

(Mexican Journal of Herpetology) (2009-2010). He is currently a full-time professor, teaching courses

in the Ecology track and in the Master’s Degree in Biological Sciences at UANL. He 1s a collaborator

of the Academy for Studies on the Richness and Conservation of Biodiversity of the FCB-UJED, and

a member of Sistema Nacional de Investigadores.

Sandra Cecilia Bocado Hernandez has been a Biologist in the Facultad de Ciencias Biologicas,

Universidad Aut6noma de Nuevo Leon, México (FCB/UANL) since 2019. With five years of

experience as a volunteer assistant in the Herpetology Laboratory of the Faculty of Biological

Sciences, her career has focused on the behavior of reptiles, and she has published three scientific

papers on this topic. In addition, she became interested in the conservation of amphibians and reptiles,

and participated in developing the Program of Action for the Conservation of the Species (PACE)

Rattlesnakes (Crotalus sp.). She currently works as a Technical Assistant in the Tamaulipas Ecoregion

and Wetlands in Pronatura Noreste, where she is involved in various conservation projects.

24 January 2022 | Volume 16 | Number 1 | e299

Amphibian & Reptile Conservation

16(1) [General Section]: 25—34 (e300).

Official journal website:

amphibian-reptile-conservation.org

Temperature-based activity estimation accurately predicts

surface activity, but not microhabitat use, in the Endangered

heliothermic lizard Gambelia sila

12.*Kathleen N. Ivey, ‘Margaret B. Cornwall, ‘Nicole Gaudenti, ‘Paul H. Maier,

3Nargol Ghazian, *Malory Owen, ‘Emmeleia Nix, *Mario Zuliani, ?Christopher J. Lortie,

‘Michael F. Westphal, and ‘Emily N. Taylor

‘Biological Sciences Department, California Polytechnic State University, San Luis Obispo, California 93401-0401, USA *Department of Biology,

University of Texas, Arlington, Texas 76019, USA *Department of Biology, York University, Toronto, Ontario M3J1P3 CANADA ‘US Bureau of

Land Management, Central Coast Field Office, Marina, California 93933, USA

Abstract.—With the existence of many endangered terrestrial ectotherms now being threatened in the face of

climate change, effective tools to aid in the management of their conservation are necessary. Temperature-based

activity estimation (TBAE) is an automated method for predicting surface activity and microhabitat use based

on the temperature of an organism and its habitat, and TBAE may be used to reduce the monitoring effort for

sensitive species. However, its efficacy has not been assessed in heliothermic species. We hypothesized that

heliothermy would facilitate the accurate prediction of surface activity due to the rapid changes in temperature

effected by exposure to solar radiation, but that TBAE would not accurately predict microhabitat use because

heliothermic lizards shuttle too frequently among microhabitats. In this study, we assessed how well ambient

air temperature and lizard physical model temperature predicted surface activity and microhabitat use of a

federally-listed Endangered lizard, Blunt-nosed Leopard Lizard, Gambelia sila, by comparing these variables to

continuously logged active lizard body temperatures in the field. While surface activity was correctly predicted

93% of the time using either ambient or physical model temperatures, the accuracy in predicting microhabitat

use only ranged from 47-72%. Finally, TBAE allowed us to predict the time of morning emergence from burrows

to within approximately 11 minutes. TBAE is a promising means for remotely monitoring surface activity and

morning emergence of heliotherms, however its utility in distinguishing microhabitat use in heliotherms is

limited.

Keywords. Automated data collection, microhabitat selection, Reptilia, surface activity, TBAE

Citation: lvey KN, Cornwall MB, Gaudenti N, Maier PH, Ghazian N, Owen M, Nix E, Zuliani M, Lortie CJ, Westphal MF, Taylor EN. 2022. Temperature-

based activity estimation accurately predicts surface activity, but not microhabitat use, in the Endangered heliothermic lizard Gambelia sila. Amphibian

& Reptile Conservation 16(1) [General Section]: 25-34 (e300).

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.

Accepted: 18 March 2021; Published: 13 February 2022

Introduction

Refugia constitute a major resource for terrestrial

organisms because they provide protection from

predators (Martin and Lopez 2004; Manicom et al. 2008)

and escape from extreme temperatures (Schwarzkopf

and Alford 1996; Polo et al. 2005), and can buffer

animals from extreme aridity and precipitation events

(Bulova 2002; Burda et al. 2007; Ivey et al. 2020).

However, essential behaviors like mate-searching

and feeding typically require surface activity, which

can be problematic for xerophilic animals due to the

especially harsh conditions they often encounter on

the surface (Martin and Pilar 1999; Krause et al. 2000;

Webb and Whiting 2005; Amo et al. 2007; Davis et al.

2008; Munguia et al. 2017). Animals that inhabit arid

environments are at risk of extinction due to the increased

temperatures and longer periods without precipitation

associated with climate change (Archer and Predick

2008; Barrows 2011), and such conditions force these

animals to seek refuge more frequently and potentially

reduce their ability to obtain resources (Buckley et al.

2015; Grimm-Seyfarth et al. 2017). Heliothermic (sun-

basking) lizards are particularly at-risk (Sinervo et al.

2010) because they already thermoregulate at high

temperatures (Cowles and Bogert 1944; Huey 1982), and

further increases in ambient temperatures will force them

into refugia. Heliothermic lizards typically have limited

plasticity in their thermal tolerance (Gunderson and

Stillman 2015). Because they are so adept at behavioral

Correspondence. “ivey.kathleen@gmail.com (Current address: Department of Biology, University of Texas, Arlington, Texas 76019, USA)

Amphib. Reptile Conserv.

February 2022 | Volume 16 | Number 1 | e300

Estimating activity and microhabitat use in Gambelia sila

thermoregulation by shuttling among the sun, shade, and

refugia, they have a low potential for adapting to higher

temperatures (Huey 1982; Huey et al. 2003; Angilletta

2009; Mufioz and Losos 2018). Therefore, heliothermic

lizards are excellent candidates for studying how shifts in

climatic events will impact organisms that rely on their

thermal environment and for understanding how we can

use temperature to model their activity.

A direct approach for studying how climate change

influences vulnerable ectotherms relies on robust methods

for collecting continuous data on body temperature and

microhabitat use, but continuous sampling of small,

heliothermic lizards is logistically challenging. Most of

these studies employ the “grab and jab” technique, in

which a lizard 1s captured, and a point sample of its body

temperature is collected using a cloacal thermometer

(Taylor et al. 2020). However, point-sampling of body

temperature is highly biased in that it provides a small

number of data points which only reflect those time

periods in which animals are active and accessible (Taylor

et al. 2004). Furthermore, tracking small individuals over

time is difficult due to limitations in radio-transmitter

size and battery life. Even in cases where telemetry

is possible, tracking these animals on a regular basis

over time presents financial and logistical challenges.

Researchers might be able to accurately predict activity

and microhabitat use based on body temperature data for

small, heliothermic lizards in arid, hot environments if

those data can be collected continuously and subjected to

robust validation. Temperature-based activity estimation

(TBAE) has been tested in two large-bodied reptiles, a

lizard and a snake (Davis et al. 2008). The use of TBAE

allowed researchers to predict surface activity 96% of

the time in the Gila Monster (Heloderma suspectum),

which forages actively on the surface, but only 66% of

the time in the Western Diamond-backed Rattlesnake

(Crotalus atrox), which tends to hide in the shade and

therefore thermoconforms more than the Gila Monster.

In this study, we investigated whether TBAE could be

used to successfully predict not just surface activity, but

also microhabitat use, in a smaller, heliothermic lizard.

Inthis study, the efficacy of TBAE for estimating surface

activity and microhabitat use was assessed in the Blunt-

nosed Leopard Lizard (Gambelia sila), a federally-listed

Endangered lizard found in a few isolated populations

in the hot and arid San Joaquin Valley and Carrizo Plain

in California, USA (IUCN 2017; Germano and Williams

2005; Germano and Rathbun 2016; Stewart et al. 2019).

Substantial financial resources are invested annually in

studying this species in order to inform management

plans for its protection and recovery. Gambelia sila

may be dramatically impacted by climate change in the

coming years (Ivey et al. 2020), although these lizards

may be able to shift their activity patterns to mitigate

warming (Germano 2019). Nevertheless, documenting

the thermal ecology and activity patterns represents an

essential component in the continued assessment and

Amphib. Reptile Conserv.

management strategy of G. sila as federal managers seek

to understand how rising temperatures, drought, and other

stressors impact lizard behavior, health, and recruitment.

We tested the hypothesis that TBAE can accurately

predict surface activity in a heliotherm, because of the

rapid change in temperature effected by exposure to solar

radiation when the lizards emerge from burrows, but

that it will be less robust in predicting microhabitat use

because heliothermic lizards shuttle frequently among

microhabitats. To test this hypothesis, we evaluated three

key predictions: (1) TBAE accurately predicts whether a

lizard is underground or surface active, (2) the accuracy

of TBAE in distinguishing microhabitat use (open

sun, shade of plant, or inside burrow) is lower than in

identifying surface activity, and (3) TBAE accurately

predicts the time of day that a lizard first emerges from

its overnight refugium.

Materials and Methods

Study Species and Sites

This study took place in the Elkhorn Plain in the Carrizo

Plain National Monument, California, USA, at two

different sites. The first site (“shrubbed”) has sparsely

distributed Ephedra californica shrubs throughout the

terrain (35.117998°, -119.629063°), while the second

site (“shrubless”) lacks Ephedra shrubs or any other

permanent ground cover and is located 6.1 km SW of

the shrubbed site (35.0891800°, -119.5750100°). The

Elkhorn Plain experiences arid summers (average high

30—40 °C) and cool winters (average low 5—9 °C, Raws

USA Climate Archive, https://raws.dri.edu/index.html,

accessed: 13 September 2019). Both sites are dominated

by Giant Kangaroo Rat (Dipodomys ingens) precincts

with extensive burrow networks that provide important

refugia for G. sila (Ivey et al. 2020). TBAE analyses of

surface activity and microhabitat use were performed

using data from the shrubbed site in 2018, and analyses

of the timing of morning emergence were performed

using data from both sites in 2019.

Adult G. sila were captured by a hand-held lasso

in early May 2018 at the shrubbed site (n = 30), and

in late April/early May 2019 at the shrubbed (” = 20)

and shrubless (n = 20) sites. Lizards were fitted with

VHF temperature-sensitive radio-transmitter collars

(Holohil model BD-2T, Holohil Systems Ltd., Carp,

Ontario, Canada) following the methods of Germano

and Rathbun (2016). The collars weighed approximately

1.5 g and never exceeded 5% of a lizard’s body mass.

We recorded standard morphometrics (mass, SVL, sex,

gravidity, and tail length), released lizards at their sites of

capture, and subsequently tracked lizards 1—3 times per

day using a VHF receiver and a Yagi antenna (R-1000

Telemetry Receiver, Communications Specialists, Inc.,

Orange, California, USA). Observations were taken daily

between 0700 and 1900 h, for a total of 147 observations

February 2022 | Volume 16 | Number 1 | e300

Ivey et al.

for all 30 lizards. During each tracking event, behavioral

observations, microhabitat description, GPS coordinates,

and a timestamp were recorded. In mid-July (the end

of their active period), the lizards were recaptured, the

radio-transmitters were removed, and the lizards were

released at their location of capture to allow estivation

for the remainder of the summer.

Body Temperature (T,)

The body temperature (T,,) of each lizard was continually

recorded (every ~5—10 minutes) as the temperature of

the radio-collar via relay to a Telonics TR-5 receiver

with a data acquisition system (Telonics Option 320)

and ~3-m tall omni antenna (Telonics model RA-6B).

The transmitters record surface temperature and not

core T,, so the recorded temperatures will change more

rapidly upon exposure to solar radiation than they would

using surgically implanted transmitters. We programmed

the system to log the interpulse intervals for each

radio-transmitter about every 10 minutes, and used the

manufacturer-provided calibration equations to convert

interpulse intervals to surface T,.

Characterizing the Thermal Habitat: Air Temperature

and Physical Models

The thermal habitat at the shrubbed site was characterized

in 2018 using two methods: air temperatures and the

temperatures of physical models. First, mean hourly data

were downloaded from the RAWS weather station at

Cochora Ranch (station ID: CXXC1), 3.7 km due east of

the shrubbed site, and used as a proxy for air temperature

(T.,,.). Second, physical models were deployed from 1-19

July 2018 (n = 6 in the sun, ” = 6 in the shade of Ephedra

shrubs, and n = 6 in burrows) following the methods of

Ivey et al. (2020). Briefly, the models consisted of 1 inch

(2.5 cm) diameter copper pipes fitted with a Thermochron

iButton (DS1921G-F5) that was programmed to record

the temperature every 10 minutes. The pipes were filled

with water and secured with PVC caps screwed onto the

male copper ends. The total length of the model was 15.3

cm. The models placed above ground were fitted with

“legs” made from copper wiring to prop them onto one

end, mimicking a basking lizard. The models placed in

burrows did not have legs.

Temperature-Based Activity Estimation (TBAE)

First, the difference between T,, and T, was used to

predict when a lizard was surface active or below ground.

When lizards are above ground, their T, often exceeds T.

as they bask in the sun, and this difference equals the

“positive temperature differential.” Positive temperature

differentials of 2, 4, 6, 8, 10, 12, and 14 °C were tested

to determine which differential best predicted when the

lizards were surface active. A solo researcher created a

Amphib. Reptile Conserv.

spreadsheet with the T, of each lizard at each of its radio-

telemetry observations along with data on its activity

(above or below ground). The “IF THEN” function in

Microsoft Excel was used to predict whether the animal

was above or below ground based on the positive

temperature differential. For example, if T, was above

T,,, by 2 °C, then the lizard was predicted to be above

ground; if not, it was predicted to be below ground.

After making the predictions, the predicted and actual

data were merged to examine how the various positive

temperature differentials impacted the accuracy of the

predictions.

Next, the temperatures of the physical models were

used to estimate microhabitat use and surface activity.

We did not use T,, because we expected the temperatures

of the physical models in the various microhabitats to be

much more relevant to these variables (Dzialowski 2005).

The average hourly temperatures of each physical model

(sun, shade, burrow) during the active hours of G. sila

(0700-1900 h) were plotted against each lizard’s T, on

the same day, and a researcher blind to the lizard’s actual

microhabitat predicted its microhabitat based on three

criteria (modified from Davis et al. 2008). (1) Lizards were

predicted to be in the open if their T, was equal to or higher

than the temperature of the models in the open. (2) Lizards

were predicted to be under shrubs if their T, was equal to or

higher than the temperature halfway between those of the

models in burrows and under shrubs, but lower than models

in the open. (3) Lizards were predicted to be in burrows if

their T, was lower than the temperature halfway between

the models in burrows and under shrubs. The predictions

of lizards in the open and under shrubs were combined

to constitute total above-ground predicted activity, and

the predictions of lizards in burrows constituted below-

ground predicted activity. Next, the blind predictions were

compared to the actual observations, and the proportions

correctly predicted were calculated. A two-proportion

Z-test in JMP (SAS Institute, Cary, North Carolina, USA,

version Pro 14) was used to compare the efficacies of the

two methods of TBAE (T,,, versus the physical models) for

predicting above- and below-ground activity.

Predicting Emergence Time

In 2019, new sets of physical models were deployed (n =

4 in the sun, = 4 in burrows) at both the shrubbed and

shrubless sites. The data for G. sila T, and the physical

model temperatures were used to predict the morning

emergence time of lizards at each site. Each day from 23

June to 14 July 2019, two lizards were randomly selected

as focal animals. Before dawn, each of two researchers

radio-tracked one focal animal and waited at least 4 m

away from the lizard’s burrow with binoculars trained on

the burrow entrance. The emergence time was recorded

in two ways: (1) the time of day when the lizard’s head

was first visible emerging from the burrow, and (2) the

time of day when the lizard’s entire body and tail had

February 2022 | Volume 16 | Number 1 | e300

Estimating activity and microhabitat use in Gambelia sila

50 5

| —@ Burrow

45 Asst hh

3 aA Ak

#0 s+se&ee+> Lizard body temperature awe ‘a a hep

35 4 A os iS .

a lee

o 30 4 Ash he debe, eouumoets Bune

Dog we et eet —— 4 ----

x : --"

EI |. Baek

>= _a

£ 204

E154

—_

10 +

0) TCT Erttt tat tt. ttt PrP dt Fett er

0730 0800 0830 0900 0930

Time of Day

Fig. 1. Methodology used to predict morning emergence time

of Gambelia sila. Emergence was predicted as the time of day

immediately preceding a distinct upward slope in the lizard’s

T, (triangles and dotted line) based on the assumption that it

would take several minutes for the radio transmitter to heat

in the sun. The rising T, was also typically associated with

the departure from the burrow physical model temperatures

(diamonds and long-dashed line) and the approach of the open

(sun) physical model temperatures (squares and short-dashed

line). In each case, the predicted time was then compared to the

observed emergence time when the lizard’s head first appeared

outside its burrow. The average difference between observed

and predicted emergence times was 11 minutes and 37 seconds.

emerged from the burrow. At the study sites, the June

and July conditions are extremely hot and arid, and

sometimes the lizards did not emerge from their burrows

at all. If a lizard did not emerge by the time T., reached

29.5 °C (since the optimum temperature for this species’

activity is from 25—35 °C; CDFW 2019), the observation

was abandoned, and that lizard was not included as a data

point. These observations took place at both the shrubbed

(n = 10 lizards) and shrubless (” = 10 lizards) sites. Two

lizards observed at the shrubbed site were too far from

the receiver array for associated T, data to be collected,

so the final sample for TBAE was 18 individual lizards

(with no repeat observations).

To predict emergence time using TBAE, a researcher

blind to a lizard’s actual emergence time plotted the lizard’s

T,, data and the lizard physical model temperatures from

that site for the duration of an emergence observation,

and then predicted the lizard’s emergence time as the time

point immediately preceding a distinct increase in the

slope of T, (Fig. 1). The predicted emergence times were

then compared with the observed emergence times, and the

absolute value of the difference in predicted and observed

emergence times (for each emergence criterion, 1.e., head

and entire body) was calculated. This value (in minutes)

represents how close the predicted emergence time was

to the actual emergence time. Observed emergence times

(minutes after sunrise) of all lizards observed (n = 20,

head only and full body) were compared between the

shrubbed and shrubless sites using a Student’s f-test, and

all data were normally distributed and had homogenous

Amphib. Reptile Conserv.

Proportion of correct prediction

2 4 6 8 10 12 14

X (where T, is at least "X" degrees greater than T,;,)

Fig. 2. Proportions of correct predictions using air temperature

to predict surface activity versus below-ground refuge use of

Gambelia sila. This method resulted in accurate predictions

64—76% of the time among the various temperature differentials

shown on the x-axis. Predictions were maximized (76%

correct) using the criterion that lizards are above ground when

their body temperatures (T,) are at least 6 °C above the air

temperature (T,, ).

variances. The sample size for head emergence was 20

and for full emergence it was 18 (since two lizards failed

to fully emerge from their burrows after 1 h).

Results

Temperature Based Activity Estimation (TBAE)

The proportions of observations of G. sila correctly

predicted to be above ground based on the criteria that

T, is at least X °C (where X = 2, 4, 6, 8, 10, 12, or 14 °C)

above T., ranged from 0.64 (for 2 °C) to 0.76 (for 6 °C).

Thus, surface versus below-ground activity was correctly

predicted 76% of the time when using the criterion that

the lizards are above ground if T, exceeds T,, by at least

6 °C (Fig. 2), and so a 6 °C differential was used in the

subsequent analysis of the efficacy of TBAE based on air

temperature for predicting surface activity.

In using TBAE to predict surface activity versus

burrow occupancy, the calculation based on T,,, (75.7%

correct overall) was superior to the calculation based on

the physical models (60.5% correct overall, Z = 3.43, p=

0.0003; Fig. 3). There was no a significant difference in

the accuracy of above-ground predictions using the two

methods, as observations predicted to be above ground

were correct about 93% of the time for both methods (Z

< 0.001, p = 1.00). A significant difference was evident

in the proportions of successful predictions for below-

ground observations, with T,, (62% correct) significantly

outperforming physical models (51% correct, Z = 1.78,

Pp = 0.037). Predicting activity based on the physical

models overestimated the time above ground specifically

by misidentifying many lizards as being in the open when

they were actually in burrows.

Of the 147 radio-telemetry fixes in 2018, 114 (77.6%)

were in burrows, 19 (12.9%) were under shrubs, and 14

(9.5%) were in the open in the sun. Figure 4 shows the

February 2022 | Volume 16 | Number 1 | e300

Ivey et al.

Above ground Below ground Above ground Below ground

Method: Air temperature Method: Biophysical model

OCorrect prediction Incorrect prediction

Fig. 3. Temperature-based activity estimation resulted in

accurate prediction of above-ground activity by Gambelia sila

more often than accurate prediction of below-ground (burrow)

occupation. Using air temperature (T.,,) was superior to using

physical model temperatures when predicting below-ground

occupation. For both methods, ~93% of observations predicted

to be above ground were correct, whereas 62% (using T,,.) and

51% (using physical models) were correct for below-ground

predictions.

relative success of predicting microhabitat use based on

the physical model data. When lizards were observed in a

given microhabitat, TBAE correctly predicted they were

in that habitat with varying accuracy (79% correct when

in the open, 47% when under shrubs, and 51% when

inside burrows).

Predicting Emergence Time

In summer 2019, lizards began emerging (head out of

burrow) at about 0745 h (with no difference between

shrubbed and shrubless sites in emergence time as

minutes after sunrise: t,, = 1.28, p = 0.22), and were fully

emerged (body and tail out of burrow) by about 0813 h

(lizards at the shrubless site tended to emerge later than

lizards at the shrubbed site: t,, = 2.11, p = 0.05, Fig. 5).

The difference between the predicted emergence and

observed emergence (head out of burrow) was 11:37

+ 01:57 (min:sec). Of the 18 observations, eight were

underestimations of predicted emergence time and

10 were overestimations. The difference between the

predicted emergence and observed full emergence was

27:00 + 02:31 (min:sec). Of the 18 observations, all

predictions underestimated the time of the full emergence

of the lizards.

Discussion

The Use of TBAE

A central goal of radio-telemetry monitoring studies

is to quantify surface activity and microhabitat use

in sensitive species like G. sila (e.g., Westphal et al.

2018), and determine how they are impacted by abiotic

conditions such as weather and biotic variables such as

prey abundance, predator behavior, and others (Germano

and Williams 2005). Here we have shown that TBAE

Amphib. Reptile Conserv.

1.0

30.9

rah

£08

‘9

< 0.7

2 0.6

9 0.5

is)

5 0.4

a 0.3

o 0.2

6 0.1

0.0

In the open Under shrub Inside burrow

OPredicted in open GPredicted under shrub @ Predicted in burrow

Fig. 4. Proportions of correctly predicted observations of

microhabitat use of Gambelia sila using temperature-based

activity estimation based on physical model temperatures.

Lizard microhabitat use was predicted correctly most often

when they were in the open, but overall microhabitat use

was not accurately predicted with TBAE in this heliothermic

lizard.

200

—

ee)

j=)

O First Emerging

SNS

Fully Emerged

Time since sunrise (min)

Shrubless Shrubbed

Site

Fig. 5. Gambelia sila emergence times (minutes after sunrise)

at two field sites (shrubless and shrubbed). Initial emergence

(head out of burrow) time did not differ between shrubbed

and shrubless sites, while lizards at the shrubbed site tended

to fully emerge (whole body and tail) earlier than lizards at

the shrubless site.

correctly estimates surface activity 93% of the time for

G. sila, a value very similar to the 96% accuracy rate

obtained for TBAE of the Gila Monster by Davis et al.

(2008). Both G. sila and the Gila Monster are active

foragers, so they are likely to be exposed to a range of

environmental temperatures as they forage, which can

alter their T, enough in comparison to their underground

refugia to facilitate TBAE. Furthermore, since G. sila

are heliothermic lizards, and because we used external

radio-transmitters, their exposure to solar radiation

should further help distinguish their surface-active T,

from their T, when inside burrows (Stevenson 1985;

Xiang et al. 1996). In contrast, TBAE failed to predict

surface activity as accurately in an ambush-foraging

rattlesnake (66% accuracy, Davis et al. 2008) because its

body temperature in the shade of its ambush site was not

sufficiently distinguishable from its body temperature

inside a refugium. Therefore, TBAE is a potentially

February 2022 | Volume 16 | Number 1 | e300

Estimating activity and microhabitat use in Gambelia sila

valuable method for researchers interested in estimating

surface activity of actively foraging species that are

expected to be exposed to relatively high temperature

variations in their environment.

The value of TBAE lies in its use of T, data that are

collected by an automated system, and therefore it does

not require direct researcher sampling. In other words,

researchers could deploy radio-transmitters on lizards to

radio-track them as needed for the goals of a particular

study, but allow TBAE to collect the data necessary

for estimating surface activity. This could significantly

save on time and resources by reducing personnel

investment in radio-telemetry. An alternate method for

collecting data on animal surface activity uses light level

geolocators, which record the intensity of blue light

(Wilmers et al. 2015) primarily as a means of tracking

migration in birds (Lisovski et al. 2019), but they can also

be externally attached to lizards (Refsnider et al. 2018) or

other terrestrial wildlife to log diel exposure to light. The

choice between TBAE and light level geolocators will

depend on the goals and budget of the study, the secrecy

and recapture rate of individuals of the species under

investigation, and other factors. One advantage of TBAE

over light level geolocators is that T, data are collected

continually by an automated receiver in TBAE, whereas

geolocator tags must be retrieved from the animals

so the data can be downloaded (Lisovski et al. 2019).

Thus, any animals lost (e.g., to predation) represent

lost data. Furthermore, in most studies on rare species

such as G. sila, researchers would probably already be

using radio-telemetry to facilitate repeated observations

of known individuals, so it would generally be simpler

and far less expensive to choose temperature-sensitive

radio-telemetry over light level geolocators. On the other

hand, light level geolocators would work very well for

recording surface activity in systems where they could

be feasibly attached to a large sample of animals with a

high recapture rate.

In this study, TBAE was not as accurate when

predicting below-ground activity. This limitation

was primarily because TBAE misidentified certain

observations as being “in the open” when the lizards

were actually in burrows. Heliothermic lizards like G.

sila maintain their T, within a narrow range, typically

within or near their laboratory-measured preferred T,

range, by shuttling between sun and shade (Lortie et al.

2015; Westphal et al. 2018; Germano 2019; Ivey et al.

2020). When a lizard moves from the sun into a burrow,

its measured T, could remain more than 6 °C above T.,.

or the physical model temperature for a short period of

time; so, if such a lizard is tracked within that period of

time, then TBAE would incorrectly assign it as being

above ground. TBAE correctly predicted below-ground

activity 62% of the time when using T,, and 51% of the

time when using the physical models.

We had expected that the physical models would

be more accurate than T, because the models were in

Amphib. Reptile Conserv.

the exact same field sites and mimic the size and shape

of lizards to facilitate realistic heat exchange with the

environment, whereas T, data merely represent the air

temperatures from a nearby weather station. The fact that

T,,, was a better estimate could be the result of several

factors. First, predictions made using T,,, have only two

possible categories: above or below ground. In contrast,

predictions made using physical model temperatures

have three categories (open, shrub, and burrow), with

open and shrub predictions then combined into above-

ground predictions. In the latter case, predicting “shrub”

use for a lizard that was actually underground because

its temperature was intermediate between the two other

options could result in overprediction of above-ground

activity; whereas if the only options were assigning it

to above or below ground, it may have been accurately

assigned as below ground. In other words, if only above

or below ground categories had been assigned using

physical models like for T,,, then the two methods

may have provided more comparable predictions.

Alternatively, the lower accuracy of the physical

models may reflect model design and radio-transmitter

construction. Our radio-transmitters were on collars and

therefore measured external temperature, not the deep T,

of the lizards, so the temperatures should change rapidly

upon exposure to the sun. In contrast, our physical

models were constructed with internal data loggers

immersed in water, which would introduce a lag time for

temperature changes due to high thermal inertia (Porter

and Gates 1969). Additionally, the Giant Kangaroo Rat

burrows used by the lizards are complicated in terms

of depth, chamber size, and soil type, likely creating a

labyrinth of thermal heterogeneity underground (Kay and

Whitford 1978), and that heterogeneity is not captured

by our physical models placed 1 m inside the burrows.

The superior performance of T,, is good news because it

means that researchers can simply download data from a

nearby weather station rather than constructing physical

models, and T, data collected from a mini weather

station deployed at the actual field site could provide

even more accurate data. In summary, TBAE using T,,.

as a reference is a highly accurate means of estimating

surface activity, but its ability to predict when lizards are

underground during daytime hours is more limited.

Microhabitat Use Predictions

To predict microhabitat use (burrow, shade, or open),

TBAE using physical models accurately predicted 79%

of the observations when the lizard was in the open (sun),

47% of the observations in the shade, and 51% of the

observations inside the burrows. Of the observations for

G. sila in the open, 100% of all predictions were above

ground (79% correctly predicted as in the sun and 21%

incorrectly predicted as under the shade of a shrub) and

in no case was a lizard predicted to be underground.

The accuracy of predictions for shade and burrows were

February 2022 | Volume 16 | Number 1 | e300

Ivey et al.

lower, probably for several reasons. First, as described

above, G. sila shuttle among these three microhabitats

regularly (Ivey et al. 2020), and an animal’s temperature

at a given radio-telemetry fix could be impacted by the

microhabitat it occupied shortly before being observed.

Second, the temperatures of the physical models in the

shade and in burrows are necessarily more similar to

each other than either is to the temperature of models

in the open that are exposed to solar radiation, so errors

in assigning shade or burrow microhabitat in TBAE

are expected (Fig. 4). Our results suggest that accurate

records of microhabitat use of heliothermic animals like

G. sila require in-person radio-telemetry, as TBAE does

not provide sufficiently accurate predictions.

The beginning of lizard emergence in the morning was

predictable to within roughly 11 minutes, which supports

the utility of TBAE as a means of remotely collecting

data on the timing of morning emergence. Lizards at

the shrubbed and shrubless sites began to emerge at

approximately the same time, and lizards at the shrubbed

site fully emerged slightly earlier in the day than lizards

at the shrubless site. In the absence of shade-providing

plants, lizards at the shrubless site may be more reliant

on the protection offered by their overnight burrows than

lizards at the shrubbed site, which can take advantage of

shrubs for thermoregulation and protection from avian

predators (Ivey et al. 2020). Lizards began emerging

from burrows at about 0745 h and were fully emerged

by 0830 h. These times agree with those reported by

Germano (2019), who compiled data on the times at

which lizards are active throughout the active season.

These data are informative for practical use by managers;

for example, California Department of Fish and Wildlife

recently revised its guidelines for G. sila protocols based

on these emergence times (CDFW 2019). As midday

temperatures increase due to climate change, lizards may

begin to emerge earlier in the morning, retreat to burrows

earlier in the afternoon, and rely more heavily on plants

for shade (Germano 2019), which could potentially

buffer G. sila from experiencing the rising temperatures.

Conducting TBAE annually would allow the testing of

this prediction with reliability and with less effort than

that required to radio-track lizards at dawn each day.

While animals must and should still be radio-tracked

to obtain data relevant to the particular question being

asked in a study and to validate TBAE and further

delineate its limitations (as we have done here), adding

TBAE to a radio-telemetry project could substantially

improve inferences about animal activity patterns and

microhabitat use while minimizing researcher effort and

expense. For example, researchers could radio-track

every other day or every third day, rather than the 2—3

times per day that is typical in studies of G. sila. To gain

further insight into how abiotic factors like ambient

temperature impact our ability to remotely predict

activity and microhabitat use, TBAE in G. sila should

be evaluated over the course of the season over multiple

Amphib. Reptile Conserv.

years. We urge researchers to consider how adopting

TBAE might augment their studies. TBAE has been

used for a variety of applications ranging from studying

maternal thermoregulation (Stahlschmidt et al. 2012)

to examining usage of artificial refugia versus natural

refugia in sympatric species (Lelievre et al. 2010). TBAE

can reduce the stress that endangered species experience

by limiting their interactions with researchers in the field.

Harnessing the power of temperature to predict animal

activity has proven to be a useful resource to augment

surveys and radio-telemetry studies, and it will assist

managers and researchers in determining how to improve

protocols for surveying and studying these species 1n the

future, while minimizing the stress imposed on these

sensitive species.

Acknowledgments.—This_ research was generously

supported by the William and Linda Frost Fund in the

California Polytechnic State University (Cal Poly),

College of Science and Mathematics, the Bureau of

Land Management, the Nature Conservancy, and the Cal

Poly Biological Sciences Department. CJL was funded

by an NSERC Discovery Grant. We are grateful to B.

Axsom of the BLM for logistical assistance and to HLS.

Butterfield of The Nature Conservancy for contributing

to the purchase of radio-transmitters in previous years.

We are also grateful to H. Smith and N. Wall for advice

on statistical analysis. Thank you to J. Hurl, B. Blom,

and E. Zaborsky who helped make this happen. We thank

the following for assistance with field work: A. Bjerre,

M. Corn, D. Deaser, N. Duong, T. Eldib, G. Espinosa, G.

Garcia, E. Gruber, S. Gonzales, J.T. Hussey, I. Ivey, M.

Kepler, C. Knowd, F. Macedo, A. Marquardt, H. Neldner,

T. Nhu, J.T. Nolan, J. Parker, K. Rock, D. Rypka, M.

Solis, T. Stratton, C. Tuskan, S. Van Middlesworth, R.

Seymour, A. Valdivia, O. Valencia Soto, and J. Whelan.

Literature Cited

Amo L, Lopez P, Martin J. 2007. Refuge use: a conflict

between avoiding predation and losing mass in lizards.

Physiology and Behavior 90: 334-343.

Angilletta MJ. 2009. Thermal Adaptation: a Theoretical

and Empirical Synthesis. Oxford University Press,

Oxford, United Kingdom. 289 p.

Archer SR, Predick KI. 2008. Climate change and

ecosystems of the Southwestern United States.

Rangelands 30: 23-28.

Barrows CW. 2011. Sensitivity to climate change for

two reptiles at the Mojave-Sonoran Desert interface.

Journal of Arid Environments 75: 629-635.

Buckley LB, Ehrenberger JC, Angilletta MJ. 2015.

Thermoregulatory behavior limits local adaptation

of thermal niches and confers sensitivity to climate

change. Functional Ecology 29: 1,038—1,047.

Bulova SJ. 2002. How temperature, humidity, and burrow

selection affect evaporative water loss in desert

February 2022 | Volume 16 | Number 1 | e300

Estimating activity and microhabitat use in Gambelia sila

tortoises. Journal of Thermal Biology 27: 175-189.

Burda H, Sumbera R, Begall S. 2007. Microclimate in

burrows of subterranean rodents — revisited. Pp. 21—

33 In: Subterranean Rodents. Editors: Begall S, Burda

H, Schleich CE. Springer, Berlin, Germany. 398 p.

California Department of Fish and Wildlife. 2019.

Approved Survey Methodology for the Blunt-nosed

Leopard Lizard. October 2019 (Revised). CDFW

BNLL protocol. California Department of Fish

and Wildlife, Sacramento, CA, USA. Available:

https://nrm.dfg.ca.gov/FileHandler.ashx? Document

ID=174900&inline [Accessed: 21 November 2019].

Cowles RB, Bogert CM. 1944. A preliminary study of the

thermal requirements of desert reptiles. Bulletin of the

American Museum of Natural History 83: 265-296.

Davis JR, Taylor EN, DeNardo DF. 2008. An automated

temperature-based option for estimating surface

activity and refuge use patterns in free-ranging

animals. Journal of Arid Environments 72: 1,414—

1,422.

Germano DJ. 2019. Activity and thermal biology of

Blunt-nosed Leopard Lizards (Gambelia sila) in the

San Joaquin Desert of California. Western North

American Naturalist 79: 428-440.

Germano DJ, Rathbun GB. 2016. Home range and

habitat use by Blunt-nosed Leopard Lizards in the

Southern San Joaquin Desert of California. Journal of

Herpetology 50: 429-434.

Germano DJ, Williams DF. 2005. Population ecology

of Blunt-Nosed Leopard Lizards in high elevation

foothill habitat. Journal of Herpetology 39: 1-18.

Grimm-Seyfarth A, Mihoub JB, Henle K. 2017. Too

hot to die? The effects of vegetation shading on past,

present, and future activity budgets of two diurnal

skinks from arid Australia. Ecology and Evolution 7:

6,803-6,813.

Gunderson AR, Stillman JH. 2015. Plasticity in thermal

tolerance has limited potential to buffer ectotherms

from global warming. Proceedings of the Royal

Society B: Biological Sciences 282: 1-18.

Huey RB. 1982. Temperature, physiology, and ecology

of reptiles. Pp. 25-91 In: Biology of the Reptilia,

Volume 12: Physiology C, Physiological Ecology.

Editors, Gans C, Pough FH. Academic Press, London,

United Kingdom. 536 p.

Huey RB, Hertz PE, Sinervo B. 2003. Behavioral drive

versus behavioral inertia in evolution: a null model

approach. American Naturalist 161: 357-366.

IUCN. 2017. Gambelia sila. The IUCN Red List of

Threatened Species 2017: e.T40690A 10336468.

Ivey KN, Cornwall M, Crowell H, Ghazian N, Owen M,

Nix E, Zuliani M, Lortie CJ, Westphal M, Taylor E.

2020. Thermal ecology of the federally Endangered

Blunt-nosed Leopard Lizard (Gambelia _ sila).

Conservation Physiology 8: coaa014.

Kay FR, Whitford WG. 1978. The burrow environment

of the Banner-Tailed Kangaroo Rat, Dipodomys

Amphib. Reptile Conserv.

spectabilis, in southcentral New Mexico. American

Midland Naturalist 99. 270-279.

Krause J, Cheng DJS, Kirkman E, Ruxton GD. 2000.

Species-specific patterns of refuge use in fish: the role

of metabolic expenditure and body length. Behaviour

137: 1,113—1,127.

Lelievre H, Blouin-Demers G, Bonnet X, Lourdais O.

2010. Thermal benefits of artificial shelters in snakes:

a radiotelemetric study of two sympatric colubrids.

Journal of Thermal Biology 35: 324-331.

Lisovski S, Bauer S, Briedis M, Davidson SC, Dhanyjal-

Adams KL, Hallworth MT, Karagicheva J, Meier

CM, Merkel B, Ouwehand J, et al. 2019. Light-level

geolocator analyses: a user’s guide. Journal of Animal

Ecology 89: 221-236.

Lortie CJ, Filazzola A, Sotomayor DA. 2015. Functional

assessment of animal interactions with shrub-

facilitation complexes: a formal synthesis and

conceptual framework. Functional Ecology 30: 41-

51.

Manicom C, Schwarzkopf L, Alford RA, Schoener

TW. 2008. Self-made shelters protect spiders from

predation. Proceedings of the National Academy of

Sciences of the United States of America 105: 14,903—

14,907.

Martin J, Lopez P. 2004. Iberian Rock Lizards (Lacerta

monticola) assess short-term changes in predation

risk level when deciding refuge use. Journal of

Comparative Psychology 118: 280-286.

Martin J, Pilar L. 1999. An experimental test of the

costs of antipredatory refuge use in the Wall Lizard,

Podarcis muralis. Oikos 84. 499-505.

Munguia P, Backwell PRY, Darnell MZ. 2017. Thermal

constraints on microhabitat selection and mating

opportunities. Animal Behaviour 123: 259-265.

Mufioz MM, Losos JB. 2018. Thermoregulatory behavior

simultaneously promotes and forestalls evolution in a

tropical lizard. The American Naturalist 191: E15—

E26.

Polo V, Lopez P, Martin J. 2005. Balancing the thermal

costs and benefits of refuge use to cope with persistent

attacks from predators: a model and an experiment

with an alpine lizard. Evolutionary Ecology 7. 23-35.

Porter WP, Gates DM. 1969. Thermodynamic equilibria

of animals with environment. Ecological Monographs

39: 227-244.

Refsnider JM, Qian SS, Streby HM, Carter SE, Clifton

IT, Siefker AD, Vazquez TK. 2018. Reciprocally

transplanted lizards along an elevational gradient

match light environment use of local lizards via

phenotypic plasticity. Functional Ecology 32: 1,227—

1,236.

Schwarzkopf L, Alford RA. 1996. Desiccation and

shelter-site use in a tropical amphibian: comparing

toads with physical models. Functional Ecology 10:

193-200.

Sinervo B, Méndez-de-la-Cruz F, Miles DB, Heulin B,

February 2022 | Volume 16 | Number 1 | e300

Ivey et al.

Bastiaans E, Cruz MVS, Lara-Resendiz R, Martinez-

Méndez N, Calderon-Espinosa ML, Meza-Lazaro

RN, et al. 2010. Erosion of lizard diversity by

climate change and altered thermal niches. Science

328: 894-899.

Stahlschmidt ZR, Shine R, Denardo DF. 2012. Temporal

and spatial complexity of maternal thermoregulation

in tropical pythons. Physiological and Biochemical

Zoology 85: 219-230.

Stevenson RD. 1985. The relative importance of

behavioral and physiological adjustments controlling

body temperature in terrestrial ectotherms. The

American Naturalist 126: 362-386.

Stewart JAE, Butterfield HS, Richmond JQ, Germano DJ,

Westphal MF, Tennant EN, Sinervo B. 2019. Habitat

restoration opportunities, climatic niche contraction,

and conservation biogeography in California’s San

Joaquin Desert. PLOS One 14: 1-18.

Taylor EN, DeNardo DF, Malawy MA. 2004. A

comparison between point- and semi-continuous

sampling for assessing body temperature in a free-

ranging ectotherm. Journal of Thermal Biology 29:

growing anthropogenic disturbances.

Amphib. Reptile Conserv.

91-96.

Taylor EN, Diele-Viegas LM, Gangloff EJ, Hall JM,

Halpern B, Massey MD, Rodder D, Rollinson N,

Spears S, Sun B, et al. 2021. The thermal ecology and

physiology of reptiles and amphibians: a user’s guide.

Journal of Experimental Zoology 335: 13-44.

Webb JK, Whiting MJ. 2005. Why don’t small snakes

bask? Juvenile Broad-headed Snakes trade thermal

benefits for safety. Oikos 110: 515-522.

Westphal MF, Noble T, Butterfield HS, Lortie CJ. 2018.

A test of desert shrub facilitation via radiotelemetric

monitoring of a diurnal lizard. Ecology and Evolution

8: 12,153-12,162.

Wilmers CC, Nickel B, Bryce CM, Smith JA, Wheat RE,

Yovovich V. 2015. The golden age of bio-logging:

how animal-borne sensors are advancing the frontiers

of ecology. Ecology 96: 1,741—1,753.

Xiang J, Weiguo D, Pingyue S. 1996. Body temperature,

thermal tolerance, and influence of temperature on

sprint speed and food assimilation in adult Grass

Lizards, Yakydromus_ septentrionalis. Journal of

Thermal Biology 21: 155-161.

Kat Ivey is a Ph.D. student at the University of Texas at Arlington (USA), where her dissertation focuses

on the population genomics of a complex system of rattlesnake species in the western United States.

Previously, she obtained her Masters of Science from California Polytechnic State University (Cal Poly),

San Luis Obispo, California, USA, where her research focused on the thermal ecology of a federally-

listed Endangered lizard species, Gambelia sila.

Margaret Cornwall is currently an undergraduate senior at California Polytechnic State University in

San Luis Obispo, California, USA. She has worked in the Physiological Ecology Reptiles Laboratory

(PERL) for four years on various aspects of data collection, data analysis, and writing. She hopes to take

her experience and passion for research into the medical field.

Nicole Gaudenti graduated with a Master of Science Degree from California Polytechnic State University,

San Luis Obispo, California, USA, studying the impacts of habitat heterogeneity on the thermal ecology

of Blunt-nosed Leopard Lizards. She obtained a B.S. in biology from the University of San Francisco in

California, USA, and has a great deal of experience studying terrestrial organisms. Prior to her Master’s

work at Cal Poly, she worked on mark-recapture studies of small mammals in the Carrizo Plain National

Monument in California. She also worked at Point Reyes Bird Observatory (Bolinas, California, USA) for

several seasons and became a certified Bander with the North American Banding Council.

Paul Maier studied Biological Sciences for his Bachelor’s Degree at California Polytechnic State

University, San Luis Obispo, California, USA, and enjoyed helping with research projects involving

snakes and lizards. He has continued conducting research in a Master’s Degree program at the same

institution, studying regenerative medicine.

Nargol Ghazian is a Master’s (soon to be Ph.D.) student at York University, Toronto, Ontario, Canada,

working under the supervision of Christopher Lortie. Her main interest lies in examining direct and indirect

interactions in arid ecosystems, with a special interest in how climatic stress influences the facilitative

effects of foundation shrubs. Currently, she is looking at ways of increasing thermal heterogeneity within

the landscape using artificial shelters with a goal of finding a practical, restorative solution to today’s ever-

February 2022 | Volume 16 | Number 1 | e300

Amphib. Reptile Conserv.

Estimating activity and microhabitat use in Gambelia sila

Mallory Owen is currently a Ph.D. student in Biology at York University (Toronto, Ontario, Canada)

supervised by Christopher Lortie. She received her Bachelor’s Degrees in Zoology and Environmental

Science from Miami University (Oxford, Ohio, USA) and her M.Sc. Degree from York University in

Biology. Her research centers on positive interactions between birds and desert foundation plants, and

the impacts that human disturbances can have on these systems. She is also a general-audience science

communicator and a grassroots advocate for environmental justice.

Emmeleia Nix graduated from California Polytechnic State University, San Luis Obispo, California,

USA with a Bachelor’s Degree in Animal Science. She has been working for the US Bureau of Land

Management since 2018, starting as one of San Diego Zoo’s “Rookies for Recovery” interns, and now

serves as a seasonal hire to support threatened and endangered species recovery efforts. She will soon be

starting her Ph.D. at University of California, Davis, California, USA.

Mario Zuliani is currently pursuing a Master’s Degree in Ecology at York University in Toronto, Ontario,

Canada. Much of Mario’s work focuses on the associations of animal species with foundational shrub

Species in desert ecosystems, such as those in the Carrizo Plain National Monument, California.

Chris Lortie is a professor of Community Ecology. He is a faculty member at York University, Toronto,

Ontario, Canada, and a senior fellow at The National Center for Ecological Analysis and Synthesis at the

University of California, Santa Barbara, California, USA. Chris studies plants and animals in deserts.

Michael Westphal received his Ph.D. in Zoology from Oregon State University, Corvallis, Oregon, USA

(Stevan J. Arnold, advisor) where he studied the evolution of color patterns in garter snakes. He was an

Ecological Genomics Postdoctoral Fellow at Kansas State University in Manhattan, Kansas, USA, where

he continued his work on color pattern genetics. Since 2008, Michael has been an Ecologist with the US

Bureau of Land Management, where he conducts research on the ecology of desert reptiles and guides the

management of endangered species.

Emily Taylor is a Professor of Biological Sciences at the California Polytechnic State University in San

Luis Obispo, California, USA, where she leads the Physiological Ecology of Reptiles Laboratory and

teaches courses in Physiology.

34 February 2022 | Volume 16 | Number 1 | e300

Official journal website:

amphibian-reptile-conservation.org

Amphibian & Reptile Conservation

16(1) [General Section]: 35-70 (e301).

Amphibians of Kokolopori: an introduction to the amphibian

fauna of the Central Congolian Lowland Forests,

Democratic Republic of the Congo

1*Gabriel Badjedjea, 7Franck M. Masudi, *Benjamin Dudu Akaibe, and ***Vaclav Gvozdik

'Department of Aquatic Ecology, Biodiversity Monitoring Center, University of Kisangani, Kisangani, DEMOCRATIC REPUBLIC OF THE

CONGO *Department of Terrestrial Ecology, Biodiversity Monitoring Center, University of Kisangani, Kisangani, DEMOCRATIC REPUBLIC

OF THE CONGO ?Faculty of Sciences, University of Kisangani, Kisangani, DEMOCRATIC REPUBLIC OF THE CONGO ‘National Museum,

Department of Zoology, Prague, CZECH REPUBLIC ° Institute of Vertebrate Biology of the Czech Academy of Sciences, Brno, CZECH REPUBLIC

Abstract.—The fauna of the Central Congolian Lowland Forests ecoregion in the Democratic Republic of the

Congo is poorly known due to the region’s remoteness and limited accessibility. An amphibian survey was

conducted in Kokolopori, including the Kokolopori Bonobo Nature Reserve, to fill this gap in our knowledge

of its amphibians. All major habitat types were surveyed using visual and acoustic encounter surveys, pitfall

and funnel trapping, and active searching during four field sessions, totaling 48 days. A total of 37 species of

anuran amphibians were recorded, while caecilians were unknown to the local human population based on

the photographs presented. Incidence-based species richness statistics estimated 37-41 amphibian species,

indicating that our survey was probably nearly complete, but we assume that some rare species or species

with secretive behaviors have probably remained overlooked. Approximately 75-80% of the total number of

species were recorded during each of the two-week portions of the fieldwork, suggesting that two weeks of

intensive surveys may have good potential for amphibian inventories in Afrotropical forests. The relatively

low number of species for this equatorial rainforest is probably a consequence of its climatic history, with the

central Congo being at a certain level of drought during the Pleistocene glaciations. The amphibian fauna is

mainly represented by forest species that inhabit the lowland forests of Central Africa. Species restricted to

intact primary forests or more euryecious species of forested ecozones were recorded. Several representatives

of genus Arthroleptis could not be assigned to any of the described species, and probably represent species

new to science. Representatives of some other genera (Leptopelis, Sclerophrys) resembled species known

from western Central Africa, but morphological differences suggested that they probably also represent new

species. The taxonomic status of Arthroleptis procterae De Witte, 1921 and Hyperolius boulengeri Laurent,

1943 are revised. Range extensions were found for several taxa, including those known only from the original

descriptions. The lack of ubiquitous synanthropic toad species of genus Sclerophrys can probably be

attributed to the well-preserved Kokolopori forests and only narrow corridors of disturbed habitat (small paths)

leading into the area. Interestingly, some degree of biofluorescence was recorded in Hyperolius phantasticus

boulengeri, which to our knowledge is the first documented case in an African anuran. Despite the relatively low

species richness, our results suggest that the Central Congolian Lowland Forests ecoregion harbors a unique

and partially endemic amphibian fauna that is to some degree differentiated from the anuran fauna to the north,

east, and west of the wide arc of the Congo River. Therefore, this survey underscores the need to protect the

central Congolian rainforests as a source of unique biodiversity, and the community-based Kokolopori Bonobo

Nature Reserve serves as a good example.

Résumé.—La faune de l’écorégion des foréts de basse altitude du Centre du Congo, en République démocratique

du Congo, est mal connue en raison de Il’eloignement de la région et de son accés difficile. Un inventaire sur

les amphibiens a ete menée a Kokolopori, y compris dans la Réserve Naturelle de Bonobo de Kokolopori,

pour combler cette lacune dans la connaissance des amphibiens. Tous les principaux types d’habitats ont ete

éechantillonnes a l’aide d’enquétes visuelles et acoustiques, de pieges a fosse et a entonnoir, et de recherches

actives au cours de quatre sessions de terrain totalisant 48 jours. Un total de 37 especes d’amphibiens

d’Anoures a ete enregistre, tandis que les Gymnophiones (cécilies) étaient inconnus de la population locale

sur la base des photographies presenteées. Les statistiques sur la richesse en espéces basées sur l’incidence

ont estimé que 37-41 espéces d’amphibiens, indiquant que notre inventaire etait probablement presque

complet, mais nous supposons que certaines espéces rares ou des especes au comportement secret n’ont

pas ete recoltées. Nous avons pu enregistrer environ 75-80% du nombre total d’espéces au cours de chacune

des deux semaines de travail sur le terrain, ce qui Ssuggére que deux semaines d’inventaire intensifs peuvent

avoir un bon potentiel pour les inventaires d’amphibiens dans les foréts afrotropicales. Le nombre relativement

faible d’especes pour la forét equatoriale humide est probablement une conséquence de histoire climatique,

Correspondence. ' *gaby.badjedjea@unikis.ac.cd, **:*vaclav.gvozdik@gmail.com

Amphib. Reptile Conserv. 35 February 2022 | Volume 16 | Number 1 | e301

Amphibians of Kokolopori, central DR Congo

le Centre Congo ayant été plus ou moins sec pendant les glaciations du Pléistocene. La faune amphibienne

est principalement représentée par des especes forestieres qui habitent les foréts de basse altitude d’Afrique

centrale. Des espéces limitées aux foréts primaires intactes ou des espeéces plus euryéciques des écozones

forestieres ont été enregistrees. Plusieurs repréesentants du genre Arthroleptis n’ont pas pu étre assignés aux

especes deécrites et representent probablement des especes nouvelles pour la science. Les representants

de certains autres genres (Leptopelis, Sclerophrys) ressemblaient a des espéces connues de l’ouest de

’Afrique centrale, mais les differences morphologiques suggéraient qu’ils repréesentaient probablement

aussi de nouvelles espéces. Le statut taxonomique d’Arthroleptis procterae De Witte, 1921 et d’Hyperolius

boulengeri Laurent, 1943 est revise. Des extensions d’aire de distribution ont ete observées pour plusieurs

taxons, y compris ceux connus uniquement par les descriptions originales. L’absence d’espeéces de crapauds

synanthropiques omniprésentes du genre Sclerophrys peut probablement étre attribuee aux foréts bien

préservées de Kokolopori et au fait que seuls d’étroits corridors d’habitats perturbes (petits chemins) menent

a la zone. Il est interessant de noter qu’un certain degré de biofluorescence a eté enregistrée chez Hyperolius

phantasticus boulengeri, ce qui, a notre connaissance, est le premier cas documente chez un Anoure africain.

Malgré la richesse relativement faible en especes, nos resultats suggérent que l’ecorégion des foréts de

basse altitude du Centre du Congo abrite une faune d’amphibiens unique et partiellement endémique, qui se

differencie dans une certaine mesure de la faune d’Anoures au nord, a l’est et a l’ouest de l’arc large du fleuve

Congo. Nous attirons donc Il’attention sur la néecessite de proteger les foréts tropicales humides du Centre du

Congo en tant que source de biodiversite unique. La Réserve Naturelle de Bonobo de Kokolopori, une réserve

communautaire, peut étre un bon exemple.

Keywords. Africa, Anura, biodiversity, checklist, Congo Basin, faunistics, frogs, herpetofauna, rainforest, species

richness

Citation: Badjedjea G, Masudi FM, Akaibe BD, GvoZzdik V. 2022. Amphibians of Kokolopori: an introduction to the amphibian fauna of the Central

Congolian Lowland Forests, Democratic Republic of the Congo. Amphibian & Reptile Conservation 16(1) [General Section]: 35—70 (e301).

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.

Accepted: 14 October 2021; Published: 14 February 2022

Introduction

The Congo Basin harbors the world’s second-largest

tropical rainforest, which is divided into several

ecoregions. The Central Congolian Lowland Forests

ecoregion is distributed south of the wide arc of the Congo

River (left bank) to approximately 3-4°S (Burgess et al.

2004; Dinerstein et al. 2017; Van de Perre et al. 2019).

The river forms a potential distribution barrier to many

species, isolating them along its northern, eastern, and

western limits (Flugel et al. 2015). This extensive area of

rainforest is deficient in data for the majority of biota, but

was predicted as a potential area of high species richness

(Burgess et al. 2004). The forests are characterized as

tropical moist terra firma broadleaf forest with little

seasonality and high humidity. Riverine forests along

major watercourses are periodically or permanently

flooded, and categorized as a separate ecoregion, the

Eastern Congolian Swamp Forests (Burgess et al. 2004;

Dinerstein et al. 2017).

Kokolopori is a region in the Djolu territory of

Tshuapa Province in the Democratic Republic of the

Congo (DRC), internationally renowned especially for

the Kokolopori Bonobo Nature Reserve (referred to

hereafter as “Kokolopori Reserve” for brevity; name in

French: Réserve Naturelle de Bonobo de Kokolopori,

abbreviated RNBK). The Kokolopori Reserve, covering

an area of approximately 4,880 km? and consisting

Amphib. Reptile Conserv.

mainly of lowland rainforest, was established in 2009

as a community-based nature reserve with the primary

aim of protecting the vital population of Bonobos (Pan

paniscus) (Almquist et al. 2010; Surbeck et al. 2017).

Following discussions with local communities, the

reserve is presently receiving a new delineation that is

targeting mostly areas of primary forests and avoiding

rural complexes. However, the new delineation has not

yet been formalized (A.L. Lokasola 2020, pers. comm. ).

The amphibian fauna of the Kokolopori Reserve is

poorly known. Unfortunately, this is a common situation

throughout the country, and especially in the remote

and difficult-to-access central areas south of the Congo

River. The road network in the area is sparse and of poor

quality. In addition to the low-quality infrastructure,

poor governance, an insufficient understanding of

the importance of research by the local population,

and difficulties with some local authorities regarding

research permissions make field research in central DRC

difficult. As a result, only limited zoological research

has been carried out in the central Congo Basin in recent

decades, mostly focusing on iconic large mammals and

birds such as Bonobo, Okapi (Okapia johnstoni), or the

Congo Peafowl (Afropavo congensis) (e.g., Mulotwa

et al. 2010; Stanton et al. 2016; Van Krunkelsven et

al. 2010). Even a large cercopithecine monkey, the

Lesula (Cercopithecus lomamiensis), was only recently

discovered and described from the central Congo Basin

February 2022 | Volume 16 | Number 1 | e301

Badjedjea et al.

(Hart et al. 2012), indicating the remoteness of the

region and limited knowledge of its biodiversity. Very

few studies have focused on the herpetofauna, with the

(central) Congo Basin being referred to as a ‘blind spot

in herpetology’ (Kielgast and Lotters 2011). Similarly,

Tolley et al. (2016) and Greenbaum (2017) pointed out

the lack of data on herpetofauna across large parts of the

Congo Basin.

Some fieldwork providing data for taxonomic,

faunistic, or conservation studies has targeted amphibians

and reptiles in various localities that are relatively distant

from the study area of this article. These include areas

from the Sankuru region (Laurent 1973, 1976a, 1979),

the vicinity of Salonga National Park (Schick et al.

2010), and other areas in the central Congo Basin (e.g.,

Chifundera Kusamba 2019; Chifundera Kusamba et al.

2014; Greenbaum et al. 2014; Gvozdik and Chifundera

Kusamba 2014; Hirschfeld et al. 2015; Kielgast et al.

2014). Two herpetological surveys have been reported

from the Lokutu area on the southern bank of the Congo

River, relatively near (approximately 120 km) our study

area. The first recorded 21 species of amphibians and 16

species of reptiles (Penner and Rédel 2007), while the

other resulted mainly in lists of expected species (only

two species of amphibians and three species of reptiles

were recorded during the week-long survey), but was

marked by apparent errors—including species that are

not expected in the Lokutu area (O’Connor 2015). The

Kokolopori Reserve was first surveyed for amphibians

by herpetologist Arne Schiotz in 2005. However, this

specialist focused mainly on the genus HAyperolius

(Schiotz 2006) and his time-limited survey targeted only

disturbed habitats at forest edges and farmbush around

villages. A second herpetological survey of this large

reserve focusing on the lizard fauna recorded 20 lizard

species in the wider reserve area, the Maringa-Lopori-

Wamba landscape (Lokasola et al. 2017). Recently, a case

of predation on the reed frog Hyperolius phantasticus

by a spider of the genus Ni/us was reported from the

reserve (Badjedjea et al. 2019). However, an overview

of the amphibians of the Kokolopori Reserve is not yet

available.

In this study, we report on the amphibians found

during four herpetological surveys conducted in the

Kokolopori Bonobo Nature Reserve and adjacent areas

in different seasons during 2018—2020, thus providing a

basis for further studies of the amphibian fauna of the

Central Congolian Lowland Forests.

Materials and Methods

Study Site

Kokolopori (Fig. 1) is a community region in the source

area of two large rivers, Lopori in the north and Luo (=

Maringa) in the south, located in the northeastern part of

Amphib. Reptile Conserv.

the Tshuapa Province, DRC. Kokolopori means ‘source

of Lopori’ in the local Longando language. It is a hilly

area with an elevation of approximately 400-500 m (with

the highest hills around 580 m), with many small streams,

often in valleys with steep slopes. The region is largely

covered by lowland rainforest of the central Congolian

type (Burgess et al. 2004; Dinerstein et al. 2017). A

substantial proportion of the forest 1s primary dense moist

forest, with areas around rural complexes characterized

as old or young secondary forests (Vancutsem et al.

2009). A large area is designated as the Kokolopori

Bonobo Nature Reserve and, together with two other

neighboring reserves (Luo Scientific Reserve, Iyondji

Bonobo Community Reserve), most of the regional

forests are formally protected. However, agricultural

and hunting pressures persist from the local human

population. The region 1s characterized by a relatively

constant humid tropical climate (rainfall present year-

round), with some oscillations in the dry (December—

February and June-August) and wet (March—May and

September-—November) seasons.

The numerous sites sampled throughout the survey

period can be grouped into four broader areas (Fig. 1),

which are named after the villages where staff of the

reserve established base camps: Yetee (0.40°N, 22.93°E,

span ~20 km), Yotemankele (0.30°N, 22.96°E, span ~7

km), Yalokole (0.22°N, 22.89°E, span ~25 km), and

after the camp site in the forest at Bechuchuu (0.48°N,

23.13°E, span ~4 km). Yalokole is located near the Luo

River, Bechuchuu is near the Lopori River (border with

Tshopo Province), and Yetee and Yotemankele are in

between the two rivers. Many small villages are present

in Kokolopori, but only the names of these four areas

are used for simplicity. All four areas are characterized

by a variety of habitats; however, primary rainforests

were surveyed mainly in Yalokole and Bechuchuu near

the Luo and Lopori rivers, respectively. The habitats

surveyed (Fig. 2) included pristine primary forests with

dense canopy cover, old and young secondary forests,

small and medium-sized streams in both forested and

rural areas, banks and flooded forests of the Luo and

Lopori rivers, and swamps and farmbush in open rural

areas. We noted the presence of small-scale gold mining

(without the use of mercury) in all four areas surveyed,

with the densest being in Bechuchuu. The pools created

by mining activities were also surveyed, as they represent

an important breeding habitat for some frogs.

Data Acquisition and Processing

Field surveys were conducted in four missions in both

the wet (May 2018, November 2018) and dry (August

2019, July 2020) seasons. Most of the field surveys

were centered around Yalokole. The last survey (July

2020) focused only on Bechuchuu, the site which had

been surveyed only marginally during the previous

three missions. Surveys were conducted for 15 days

February 2022 | Volume 16 | Number 1 | e301

Amphibians of Kokolopori, central DR Congo

Kokolopo: H

Bonobo

Nature

Fig. 1. Map of the study site. (A) Democratic Republic of the Congo (DRC), Congo Basin (red line) with the Congo River

(blue line), and Kokolopori (yellow rectangle). Vegetation types follow Vancutsem et al. (2009): greenish tones correspond to

forests, brownish tones to open areas; within Kokolopori (inset in left corner), dark green corresponds to primary dense moist

forests, lighter green to old and young secondary forests, and yellow-white to rural and agricultural complexes. (B) Kokolopori

Bonobo Nature Reserve (yellow line) as delineated in 2009 on a satellite map corresponding to 2005—2010 (FACET 2010), with

collecting sites (red dots). White stars denote four sampling areas (base camps). Kokolopori is a spring area of two large rivers,

Lopori and Luo (= Maringa).

Amphib. Reptile Conserv. 38 February 2022 | Volume 16 | Number 1 | e301

Badjedjea et al.

Aa Sie TE z ‘ ee "ee ed * .- oe

Fig. 2. Habitats of Kokolopori. (A) Rural complexes are common along pathways (e.g., habitat of Ptychadena sp. aff. mascareniensis

when puddles are formed after rains). (B) Herbaceous habitat and shrubs along a small stream in disturbed forest (e.g., Hyperolius

phantasticus boulengeri, Congolius robustus, Leptopelis ocellatus schiotzi, and Ptychadena perreti). (C) Disturbed primary forest

(e.g., Arthroleptis phrynoides, and A. tuberosus procterae). (D) Small stream in primary dense moist forest (e.g., Ptychadena

aequiplicata, Sclerophrys cf. funerea, and S. sp. aff. camerunensis 2). (E) Primary forest along the Luo (= Maringa) River. (F)

Flooded forest near the Luo River (e.g., Aubria masako, Hymenochirus cf. boettgeri, and Xenopus pygmaeus). (G—H) Bonobo (Pan

paniscus), the umbrella species of the Kokolopori Bonobo Nature Reserve facilitating the conservation of the whole ecosystem.

Amphib. Reptile Conserv. 39 February 2022 | Volume 16 | Number 1 | e301

Amphibians of Kokolopori, central DR Congo

per mission, with the exception of the last survey (three

days), by two experienced herpetologists (G.B. and either

F.M.M. or V.G.) plus two to four local guides. Amphibians

were detected during nocturnal and, to a limited extent,

diurnal visual and acoustic encounter surveys, as well as

active searches under logs and stones, or in holes, and

in both natural and artificial pools. All specimens were

hand-captured, except for the tadpoles and pipid frogs

which were collected either by dip netting and dredging

in pools and streams, or by funnel traps. In May 2018,

three lines of pitfall traps were installed in the primary

and secondary forests around Yalokole. The pitfall traps

were deployed on 100 m long transects consisting of 20

traps placed at 5 m intervals (Nicolas et al. 2003). Plastic

buckets (10 L) were buried in the ground, with their upper

edges divided in half by a drift-fence barrier (consisting

of side-by-side pieces of tarpaulin, cut evenly to a height

of 50 cm) passing through their center. At each end of

the line, there was a 2.5 m extension of the fence from

each end bucket. The pitfall traps were left in place for 15

days and were checked three times per day. Photographs

of African caecilians were presented to the local people

to ascertain whether they might be present. At night, we

also investigated the possible fluorescence emission of

anurans using a handheld UV-flashlight (LED, Esco Lite

51 UV Black Light) with a peak wavelength range of

395—400 nm.

All geographic locations were recorded with a

handheld GPS device. Individuals were photographed

either in situ, or later arranged usually on leaves, and

in standard positions with a scale and tag. Vouchered

specimens were fixed in 4—10% formalin or 96% ethanol,

and later stored in 70-75% ethanol. Tissue samples were

preserved in 96% ethanol. Specimens are deposited in the

herpetological collections of the Biodiversity Monitoring

Center (Centre de Surveillance de la Buiodiversité;

CSB:Herp); University of Kisangani, DRC (field series

RNBK); National Museum, Department of Zoology,

Prague, Czech Republic (NMP-P6V); and the Institute

of Vertebrate Biology of the Czech Academy of Sciences

([VB-H), Research Facility Studenec, Brno, Czech

Republic (field series CD—and this material will later be

transferred to NMP-P6V).

There is currently no key to species identification for

the central Congolian herpetofauna. Therefore, species

identifications were based on comparisons to other

reference specimens from DRC and other Central African

countries (Cameroon, Central African Republic, Congo

Republic, and Gabon), and to type specimens deposited

in the Royal Museum for Central Africa, Tervuren

(Belgium, RMCA); National Museum of Natural

History, Paris (France, MNHN); American Museum of

Natural History, New York (USA, AMNH); Museum of

Comparative Zoology, Harvard University, Cambridge,

Massachusetts (USA, MCZ); Natural History Museum,

Berlin (Germany, ZMB); and Natural History Museum,

London (UK, BMNH). The first taxonomic orientation

Amphib. Reptile Conserv.

was based on the key for Gabon and Mbini (Frétey et

al. 2011) and two treefrog books (Amiet 2012; Schietz

1999), and the identification of Hyperolius was critically

compared with Schigtz (2006). Where the available

literature suggests that a particular taxon represents

a Species complex or the taxonomy is unclear, we use

the term confer (cf.) in our identifications. Similarly,

where we assume that the taxon under discussion is

likely to represent a previously undescribed species

similar to a known one, we use the term species affinis

(sp. aff.). Where appropriate, we also assign subspecies

identifications to assess morphological and genetic

(unpublished data) differences. Genetic data will be

published in separate taxonomic studies in the future.

We anticipate that where subspecies identifications are

applied, these taxa may later be elevated to full species

after targeted taxonomic revisions. For now, however,

we prefer to remain conservative and use subspecies

names in accordance with recent recommendations by

Hillis (2019, 2020) and de Queiroz (2020). Those authors

have concluded that the subspecies category 1s useful for

distinguishing geographic races and naming geographic

variations whenever such distinctions are important or

practical. Since the amphibian fauna of the central Congo

Basin shows some differences from the fauna of the rest

of Central Africa, we consider it practical to distinguish

these taxa at least at the subspecies level in this article.

Species richness was assessed as species accumulation

curves based on the cumulative number of species

observed and using incidence-based statistical estimates

with 100 randomizations in EstimateS software (Colwell

2013; Colwell and Elsensohn 2014). Analyses were

conducted for each of three 15-day missions, 1.e., two

surveys during the wet season (May 2018, November

2018) and one during the dry season (August 2019), and

for a cumulative survey covering a total of 48 days. The

values of several estimators were compared (Table 1):

first-order Jackknife (Jackknife 1; Burnham and Overton

1978, 1979), Chao 2 with bias correction (Chao 1987),

Incidence Coverage-based Estimator (ICE; Chao et

al. 1992: Chazdon et al. 1998), and Bootstrap richness

estimator (Smith and van Belle 1984). Two estimators

were plotted, Jackknife 1 and Chao 2, and they have been

rated as generally performing well (e.g., Basualdo 2011;

Williams et al. 2007) (Fig. 3).

Results

A total of 37 species of anuran amphibians from nine

families and 16 genera were recorded (Table 2). According

to the photographs presented to the local inhabitants, they

did not recognize caecilians, suggesting that this group

of amphibians is absent from Kokolopori. The most

frequently encountered taxa were the scansorial (arboreal)

Leptopelis ocellatus schiotzi(Arthroleptidae; encountered

on 33 of 48 survey days), Amnirana cf. albolabris

(Ranidae; 31), Congolius robustus (Hyperoliidae; 28),

February 2022 | Volume 16 | Number 1 | e301

Badjedjea et al.

Table 1. Species richness of amphibians in Kokolopori, observed and estimated using four statistical estimators: first-order

Jackknife, Chao 2 with bias correction, Incidence Coverage-based Estimator (ICE), and Bootstrap richness estimator. Means,

standard deviations, and 95% confidence intervals (Chao 2) are reported. Cumulative results are in bold.

Number Number of species estimated

of species

Field session Days observed Jackknife 1 Chao 2 ICE Bootstrap

Wet season | 15 31 42.2+2.8 39.8 +68 46.2+3.1 36.1 + 1.0

(33.3-64.7)

Wet season 2 15 30 38.4 + 3.0 36.7+6.1 37.04+2.4 33.9412

(31.5-60.4)

Dry season 15 27 32.6419 29.3 42.7 30.5+41.4 29.8 +0.7

(27.4—41.4)

Cumulative 48 37 39.0 + 2.0 37.2 + 0.6 37.6 + 0.6 38.5 + 0.6

(37.0—-41.0)

and Hyperolius phantasticus (Hyperoliidae, 24). In terms Species Accounts

of abundance, the most abundant species on average was

H. phantasticus, which was commonly found in the wet

season. However, its abundance was not so evident in the

dry season. The overall composition of the most frequently

encountered taxa during the wet and dry seasons was the

same, except that Leptopelis christyi (Arthroleptidae) was

more commonly encountered during the dry season (8

out of 18 days) than during the wet season (4/30 days),

and the aforementioned H. phantasticus was less visible

during the dry season (5/18 days in the dry season vs.

19/30 days in the wet season). Within primary forest, the

leaf-litter frog Phrynobatrachus cf. giorgii was one of

the most commonly encountered species, while within

degraded habitats, particularly farmbush, Amnirana

cf. albolabris was the most common. Along small

watercourses, hyperoliid treefrogs were most common.

The four surveyed areas did not appear to differ in species

composition, while some differences could be observed

among habitats. Biofluorescence upon illumination with a

UV-light of wavelength of 395-400 nm was observed and

documented in one species, H. phantasticus (see Species

Accounts), while the approximately 15 other species that

were tested (mostly hyperoliids) did not exhibit obvious

fluorescence.

Species Richness

During each of the 15-day field sessions, 73-84% of the

total number of species were recorded (observed and

estimated). The numbers of species observed during these

three missions were 31 (wet season), 30 (wet), and 27

(dry). Species richness estimates were similar for the two

wet season missions and the cumulative survey (Table 1,

Fig. 3A—B,D), but standard deviations and 95% confidence

intervals were much higher/wider for the 15-day sessions

than for the cumulative 48-day survey. A lower number

of species was estimated based on the dry season mission

(Table 1, Fig. 3C). In the case of the cumulative survey, the

numbers of species observed and statistically estimated

converged (Fig. 3D), with estimates of 37-41 species

(Chao 2, 95% confidence interval).

Amphib. Reptile Conserv.

Amphibia: Anura

Arthroleptidae

Arthroleptis Smith, 1849

Five species of Arthroleptis were found, one of a large

size, One medium-sized, and three rather small-sized

(as described below). The latter three correspond

morphologically to the presently invalid genus

Schoutedenella De Witte, 1921. The phylogenetic

diversity of Arthroleptis from the central Congo Basin

is the subject of a separate study (V. Gvozdik et al., in

prep.).

Arthroleptis tuberosus Andersson, 1905

Arthroleptis tuberosus procterae De Witte, 1921 (new

status)

Fig. 4C.

Area: All.

Season/survey: Wet (May, Nov 2018), dry (Aug 2019).

Material: CSB:Herp:RNBK 031, 056, 076, 085, 125,

216, 545, 585, 706, 719, 734; IVB-H-CD 18241, 18242,

18265, 18266, 18267.

Comments: Arthroleptis tuberosus is a medium-sized and

little-known species, but it is supposedly widespread from

Cameroon (or eastern Nigeria; Nneji et al. 2019) to eastern

DRC (IUCN SSC Amphibian Specialist Group 2017).

Given that Amiet and Goutte (2017) recently proposed

that this species consists of multiple subspecies (4. ¢

tuberosus, A. t. adelphus Perret 1966), and given the type

localities of the available names, the central and eastern

Congolian population should bear the nomen A. tuberosus

procterae De Witte, 1921 (new status), type locality: “Beni

(Kivu).” However, the status of this taxon, particularly

whether it eventually merits full species status, requires

further research. Arthroleptis tuberosus is abundant in

forested regions in central, northern, and eastern DRC (G.

Badjedjea, unpub. obs.). In Kokolopori, A. tuberosus was

mostly found in forests, both primary and disturbed, and

usually at night. Some specimens were caught in the early

February 2022 | Volume 16 | Number 1 | e301

Amphibians of Kokolopori, central DR Congo

Table 2. Species diversity of amphibians found in Kokolopori (2018—2020).

Family Taxon

Arthroleptidae Arthroleptis tuberosus procterae

Arthroleptis sp. aff. variabilis

Arthroleptis phrynoides

Arthroleptis sp. aff. phrynoides

Arthroleptis sp. aff. xenochirus

Cardioglossa congolia

Leptopelis calcaratus meridionalis

Leptopelis christyi

Leptopelis ocellatus schiotzi

Bufonidae Sclerophrys cf. funerea

Sclerophrys sp. aff. camerunensis |

Sclerophrys sp. aff. camerunensis 2

Hyperoliidae Afrixalus equatorialis

Afrixalus osorioi

Afrixalus cf. quadrivittatus

Congolius robustus

Cryptothylax greshoffii

Hyperolius cf. cinnamomeoventris

Hyperolius cf. langi

Hyperolius cf. platyceps

Hyperolius ocellatus purpurescens

Hyperolius phantasticus boulengeri

Hylambates verrucosus

Phrynobatrachidae Phrynobatrachus cf. auritus

Phrynobatrachus cf. giorgii

Phrynobatrachus sp. aff. auritus

Pipidae Hymenochirus cf. boettgeri

Hymenochirus cf. boulengeri

Xenopus pygmaeus

Ptychadenidae Ptychadena aequiplicata

Ptychadena christyi

Ptychadena perreti

Ptychadena sp. aff. mascareniensis

Pyxicephalidae Aubria masako

Ranidae Amnirana cf. albolabris

Amnirana lepus

Rhacophoridae Chiromantis cf. rufescens

morning. This species usually sits on low vegetation at

around 0.5—1 m high, and it is occasionally found in leaf-

litter. It was often found after rain.

Arthroleptis sp. aff. variabilis

Fig. 4E.

Area: Yalokole, Yetee, Yotemankele.

Season/survey: Wet (May, Nov 2018), dry (Aug 2019).

Amphib. Reptile Conserv.

Notes

Possibly a separate species

Probably an undescribed species

Possibly a separate species

Probably an undescribed species

Possibly a separate species

Probably an undescribed species

Material: CSB:Herp:RNBK 022, 023, 025, 087, 547;

IVB-H-CD 18371-18375.

Comments: A large Arthroleptis that is widespread in

the central and northeastern DRC, although specimens

from Kokolopori seem to be larger than those from other

regions. It resembles A. variabilis Matschie, 1893 (type

locality in southwestern Cameroon) but is stouter, and

superficially similar to A. stenodactylus Pfeffer, 1893

February 2022 | Volume 16 | Number 1 | e301

Badjedjea et al.

Wet season (May 2018)

40+A B

Number of species

0 5 10 15 20

Wet season (Nov 2018)

50 i a ee vat a, Sa

Dry season (Aug 2019)

10 15 5 10 15

eeete

. e%

fen

weet et es cess,

Jackknife 1

Chao 2

Chao 2 (95% Cl)

Species observed

25 30 35 40 45

Number of days

Fig. 3. Species richness of amphibians in Kokolopori (species accumulation curves) as based on the number of species observed

(black) and statistically estimated using the Jackknife 1 (blue) and Chao 2 (red) methods. (A) Wet season, May 2018. (B) Wet

season, November 2018. (C) Dry season, August 2019. (D) Cumulative data from the entire period, 2018-2020 (48 days). Red

dotted lines show the 95% confidence interval for Chao 2.

from the savannas of the southeast. It seems to be mostly

terrestrial, typically found jumping on the ground during

the morning hours. Individuals of this species were mainly

collected in the primary forest habitat. The individuals

exhibited variable coloration from light to dark brown. A

phylogeographic study of the A. variabilis complex will

shed more light on the evolutionary history and taxonomy

of these Squeaker Frogs (D.C. Blackburn et al., in prep.).

Small-sized “Schoutedenella”

Arthroleptis phrynoides (Laurent, 1976)

Fig. 4A—B.

Area: Yalokole.

Season/survey: Wet (May 2018).

Material: CSB: Herp:RNBK 058, 124.

Comments: Originally described in the genus

Schoutedenella (Laurent 1976a). Only two probably

adult females were found during the whole study,

suggesting that this species is probably very cryptic or

rare. This also corresponds to the relatively late discovery

and description of this species, which was based only on

one male and one female (Laurent 1976a). The external

Amphib. Reptile Conserv.

morphology is concordant to the type material. Both

specimens were found in primary forest near a stream

(two different streams) sitting on vegetation about 0.5

m high in May 2018. The name phrynoides refers to the

resemblance to the bufonid arboreal genus Nectophryne

(Laurent 1976a). The similar morphology could point

to an arboreal life history of this Arthroleptis, possibly

also explaining its rarity. The species has a conspicuous

verrucosity, the throat is black in both females, and the

anterior part of the venter is black with diffused white

spots. The tips of the toes and fingers are enlarged

into distinct discs, an obvious adaptation to scansorial

life. Our record is probably the first finding of this

Species since its description, extending its known range

approximately 250 km northward from the type locality

in Sankuru Province. The two localities are similar in

terms of vegetation and climate, because they occur

within a continuous forest block.

Arthroleptis sp. aff. phrynoides

Fig. 4D.

Area: Bechuchuu, Yalokole.

February 2022 | Volume 16 | Number 1 | e301

Amphibians of Kokolopori, central DR Congo

= a - —

= =

Fig. 4. Arthroleptids, Arthroleptis and Cardioglossa. (A) Arthroleptis phrynoides (female). (B) A. phrynoides, ventral view (same

specimen as in A). (C) A. tuberosus procterae (female). (D) A. sp. aff. phrynoides (female). (E) A. sp. aff. variabilis (male). (F) A.

sp. aff. xenochirus (male). (G) Cardioglossa congolia (male). (H) C. congolia, ventral view (same specimen as in G).

Amphib. Reptile Conserv. 44 February 2022 | Volume 16 | Number 1 | e301

Badjedjea et al.

Season/survey: Wet (Nov 2018), dry (Aug 2019).

Material: CSB:Herp:RNBK 704; IVB-H-CD 18145,

18240.

Comments: This is another species that is either rare

or has cryptic ecology, because only two females and

one juvenile were found. It resembles A. phrynoides,

which is syntopic, but has substantial differences in both

morphological features and color pattern. Conspicuous

markings in the form of white-black marbling are present

on the ventrum and contrast with the reddish underside

of the hind limbs. Discs on toes and fingers are not as

distinct as they are in A. phrynoides, suggesting a less

scansorial ecology. The two specimens were found on

low herbaceous vegetation about 0.5 m high in disturbed

habitat at the forest margin, while one specimen was

hopping on the ground in primary forest at night after

almost a full day of rain. The morphology does not

conform to any of the known described species. A formal

description of this new species will be given elsewhere

(V. Gvozdik et al., in prep.).

Arthroleptis sp. aff. xenochirus

Fig. 4F.

Area: Yalokole.

Season/survey: Wet (May 2018).

Material: CSB: Herp:RNBK 054, 140.

Comments: Another small Arthroleptis from the

“Schoutedenella” group, like A. phrynoides and A. sp.

aff. phrynoides, although this one rather resembles A.

xenochirus from wooded savanna uplands of southern

DRC and surrounding countries (Bittencourt-Silva 2019;

Ernst et al. 2020). The vegetation and climate in central

Congo are very different from the southern part of the

country (Vancutsem et al. 2009), suggesting that this

Arthroleptis is probably another yet-undescribed species

deserving more attention.

Cardioglossa Boulenger, 1900

This genus is related to Arthroleptis and was recently

synonymized with it within a large phylogenetic meta-

analysis of recent amphibians (Dubois et al. 2021;

given obviously in error as a synonym of Astylosternus

in the text, while as a synonym of Arthroleptis in the

phylogenetic trees). However, we follow the last focused

study on these genera (Blackburn 2008) and continue

to recognize them here as two separate genera, as do

Blackburn et al. (2021) in the note at the end of the article

on the phylogeny of Cardioglossa and Frost (2021) in his

database.

Cardioglossa congolia Hirschfeld, Blackburn,

Greenbaum, and Rédel, 2015

Fig. 4G—-H.

Area: Bechuchuu, Yalokole.

Season/survey: Wet (May, Nov 2018), dry (Aug 2019,

Jul 2020).

Amphib. Reptile Conserv.

Material: CSB:Herp:RNBK 143, 240, 717, 854; IVB-H-

CD 18193, 18194, 18264.

Comments: A little-known Cardioglossa from the Central

Congolian forests, closely related and morphologically

similar to C. gratiosa from northwestern Central Africa

(Blackburn et al. 2021). This species is known only from

males so far (Hirschfeld et al. 2015). We also found

only males, calling from leaf-litter near small streams

in primary forest. This species has obviously cryptic

ecology, and it is not easy to locate even calling males,

while females remain unknown to science. Our new

records from Kokolopori are located between the only

two known distribution sites of this species in western

Mai-Ndombe Province and western Tshopo Province

(Hirschfeld et al. 2015).

Leptopelis Gunther, 1859

Three species of Leptopelis were recorded here. Two of

them are identifiable to taxa described at the subspecies

level. However, considering their morphological

distinctions and geographic distance from the type

localities of the nominotypical subspecies, they may

represent full species.

Leptopelis calcaratus (Boulenger, 1906)

Leptopelis calcaratus meridionalis Laurent, 1973

Fig. SA-B.

Area: Yalokole, Yotemankele.

Season/survey: Wet (May 2018), dry (Aug 2019).

Material: CSB:Herp:RNBK 144, 541, 542.

Comments: This taxon has not been reported since its

description. Here, three males were found in primary

forest on tree branches (4 m high in one case) several

meters from streams. Two color morphs were recorded,

two males were brown (Fig. 5A) and one was contrasting

orange-black (Fig. 5B). It is worth noting that a similar

orange morph was documented by Amiet (2012) as a

rare coloration in L. calcaratus from Cameroon. These

treefrogs were identified based on a comparison with

the type material of L. calcaratus meridionalis. This

subspecies was described as distributed south of the

wide Congo River arc, while the populations to the

east, north, and west were identified as belonging to

the nominotypical subspecies (Laurent 1973). This

taxon morphologically differs in several features, most

importantly by a less developed spur on the heel and more

extensive foot webbing. The level of distinction of this

central Congolian taxon needs to be further investigated

as it may represent a full species.

Leptopelis christyi (Boulenger, 1912)

Fig. SC—E.

Area: Bechuchuu, Yalokole, Yotemankele.

Season/survey: Wet (May 2018, Nov 2018), dry (Aug

2019, Jul 2020).

Material: CSB:Herp:RNBK 171, 183, 227, 242, 243,

February 2022 | Volume 16 | Number 1 | e301

Amphibians of Kokolopori, central DR Congo

Fig. 5. Arthroleptids, Leptopelis. (A) L. calcaratus meridionalis, brown morph (male). (B) L. calcaratus meridionalis, orange-black

morph (male). (C) L. christyi, brown morph (male). (D) L. christyi, green morph (male). (E) L. christyi (juvenile). (F) L. ocellatus

schiotzi (amplectant pair). (G) L. ocellatus schiotzi (male). (H) L. ocellatus schiotzi (female).

Amphib. Reptile Conserv. February 2022 | Volume 16 | Number 1 | e301

Badjedjea et al.

250, 252, 270, 271, 293, 586, 622, 638, 644, 763, 800—

802, 808, 809, 812, 813; IVB-H-CD 18341.

Comments: This treefrog is locally common (ce.g.,

found in flooded forest) but missing or difficult to detect

in other places. Two color morphs were recorded: the

more common brown morph (Fig. 5C), and the brown-

green morph (with a green dorsal triangle, stripes on

limbs, and other smaller markings) which is rarely

almost completely green (Fig. 5D). Juveniles are bright

green with yellowish joints on the limbs (Fig. 5E).

Interestingly, only a single specimen (metamorph) was

found during the survey in November, when no adults

were found. The metamorph was perched about | m high

on herbaceous vegetation along a larger stream, together

with tens of metamorphs of L. ocellatus schiotzi. This

species was described from a forested region in Uganda

and has been considered to be an eastern Central African

element (e.g., Channing and Rodel 2019; Schigtz 1999).

However, based on accumulating evidence (e.g., Amiet

2012; Dewynter and Frétey 2019; V. Gvozdik, unpub.

obs.), it seems that this treefrog 1s widespread in Central

Africa from Cameroon and Gabon across the forested

zone of the Congo Basin, and marginally to East Africa.

Its presence in Angola 1s also possible.

Leptopelis ocellatus (Mocquard, 1902)

Leptopelis ocellatus schiotzi Laurent, 1973

Fig. SF—-H.

Area: Yalokole, Yetee, Yotemankele.

Season/survey: Wet (May 2018, Nov 2018), dry (Aug

2019).

Material: CSB:Herp:RNBK 007, 012, 018, 028, 035,

037, 041, 071, 078, 089, 121, 126, 138, 144, 158, 239,

268, 285, 294, 380, 415, 417, 418, 420, 421, 423, 442,

506, 521; IVB-H-CD 18085, 18086, 18146, 18161,

18168, 18169, 18252, 18258-18262, 18295, 18296,

18334-18337, 18340, 18388, 18389.

Comments: Like L. calcaratus meridionalis, this

taxon has not been reported since its description. It is

supposedly endemic to the Central Congolian forests.

In comparison to the nominotypical subspecies, which

occurs in western Central Africa, L. ocellatus schiotzi

is more robust and has a relatively shorter and wider

head (Laurent 1973). The degree of differentiation of

these two taxa needs to be thoroughly investigated

to determine whether the central Congolian taxon

may represent a separate species. The sexual size

dimorphism is conspicuous, with females much larger

than males (Fig. 5F). This treefrog seems to be one of

the most common species, occurring in a wide range of

habitats, most often in disturbed habitats or ecotones

between forest and open areas near streams. Males

often call from high positions.

Bufonidae

Amphib. Reptile Conserv.

Sclerophrys Tschudi, 1838

This genus was formerly known as Bufo or

Amietophrynus, but it is now named Sclerophrys (Ohler

and Dubois 2016). We were unable to identify any of

the three species of Sclerophrys toads to a described

Species with certainty, and probably at least two of them

represent undescribed species. All three seem to be

confined to forested habitats. Interestingly, none of the

typical and usually common (sem1-) synanthropic species

of the Congo (S. gutturalis, S. pusilla, S. regularis)

was recorded in this survey. Sclerophrys toads from

the forests of Central Africa form a clade containing S.

camerunensis, S. funerea, S. gracilipes, S. kisoloensis, S.

villiersi, and several undescribed species (Liedtke et al.

2016, 2017), which for simplicity we tentatively name as

the S. camerunensis-gracilipes group, and are in need of

taxonomic revision (E. Greenbaum et al., in prep.).

Sclerophrys cf. funerea (Bocage, 1866)

Fig. 6A-C.

Area: Bechuchuu, Yalokole, Yotemankele.

Season/survey: Wet (May 2018, Nov 2018), dry (Aug

2019, Jul 2020).

Material: CSB:Herp:RNBK 116, 188, 204, 211, 276,

540, 570, 690, 711, 721, 832; IVB-H-CD 18313.

Comments: This species was usually found in or near

forest streams in several sites during all surveys. However,

it was rather uncommon. Calling was heard in the early

morning. Its coloration was usually dark to blackish in

males, brownish in females, and often with some reddish

patches in both sexes. Parotoid glands are rather indistinct.

Verrucosity is relatively strong, especially on limbs and

the posterior part of the dorsum. However, the dorsum

of males is smoother with flattened warts during the

mating period (Fig. 6A). Sclerophrys funerea is a little-

known species despite its presumed large distribution

range covering most of the Congo Basin, countries of the

Albertine Rift, and most of Angola where the type locality

is in the north of the country (Marques et al. 2018). It is

probably absent from Cameroon (J.-L. Perret sensu Joger

1982) and Equatorial Guinea (Sanchez-Vialas et al. 2020),

and is not reliably documented (at least from published

photographs) from Gabon (Dewynter and Frétey 2019;

Pauwels and Rodel 2007), and probably not even from the

Republic of the Congo, as S. gracilipes has probably been

confused with this species. Our specimens from Kokolopori

correspond morphologically to specimens at RMCA

identified as Bufo funereus funereus by J. Hulselmans,

who described Bufo funereus djohongensis from

Cameroon (Hulselmans 1977), which was later elevated

to full species (Joger 1982), although it is possibly a junior

synonym of S. villiersi (Frétey et al. 2011; Channing and

Rodel 2019). Since the holotype of S. funerea was most

likely destroyed in a 1978 fire, the photograph in Perret

(1976a) is probably the only visual information on the

holotype (and the quality of the specimen was rather

February 2022 | Volume 16 | Number 1 | e301

Amphibians of Kokolopori, central DR Congo

Fig. 6. Bufonids, Sclerophrys. (A) S. cf. funerea, dorsal view (male in breeding condition, with smoother dorsal skin).

(B) S. cf. funerea, brown morph (female). (C) S. cf. funerea, black morph (male). (D) S. sp. aff. camerunensis 1, with

distinct coloration (subadult). (E) S. sp. aff. camerunensis | (male). (F) S. sp. aff. camerunensis | (female). (G) S. sp. aff.

camerunensis 2 (male). (H) S. sp. aff. camerunensis 2 (female).

Amphib. Reptile Conserv. February 2022 | Volume 16 | Number 1 | e301

Badjedjea et al.

poor). There are other names available (Bufo benguelensis

Boulenger, 1882; Bufo buchneri Peters, 1882; Bufo

decorsei Mocquard, 1903; Bufo berghei Laurent, 1950;

Bufo latifrons mayombensis Hulselmans, 1977) that may

or may not be synonyms of S. funerea (Tandy and Keith

1972). Comparing our material from Kokolopori with the

holotype of S. buchneri, for which the type locality is also in

northeastern Angola (about 500 km from the type locality

of S. funerea), we found no significant differences. This is

further indirect evidence that S. buchneri may represent

a junior synonym of S. funerea, as already suggested by

Tandy and Keith (1972). However, a thorough revision

of the S. camerunensis-gracilipes group is needed, and a

designation of a neotype of S. funerea will probably be

necessary to stabilize the taxonomy of Central African

forest toads. In view of the above uncertainties, and given

the distance of about 1,300 km from the type locality

(which is located in a different ecoregion), we refer to our

population as S. cf. funerea for now.

Sclerophrys sp. aff. camerunensis 1

Fig. 6D-F.

Area: Bechuchuu, Yalokole, Yetee.

Season/survey: Wet (May 2018, Nov 2018), dry (Aug

2019, Jul 2020).

Material: CSB:Herp:RNBK 019, 114, 139, 232, 244,

246, 247, 249, 258, 432, 522, 523, 527, 528, 705, 707,

740, 866-868; IVB-H-CD 18176, 18177, 18244, 18253,

18254, 18268, 18327-18333, 18377.

Comments: This and the following species have a

very similar general morphology. However, they differ

consistently in the size of the tympanum (see below)

and probably other morphological features. Both species

belong to the S. camerunensis-gracilipes group, with

males being slender and rather resembling S. gracilipes,

while the more robust and colorful females resemble

S. camerunensis. This species, which we provisionally

name “aff. camerunensis 1,” appears to be more common,

usually occurring near streams. Male coloration varies

from yellowish to dark brown, with females usually

having amore contrasting dorsal color pattern and yellow-

reddish body sides. The dorsal stripe may be visible but

sometimes it is not present. Breeding was recorded in

a small shallow river where males were hiding among

aquatic plants. A similar, possibly undescribed species

was reported from northern Angola (Ernst et al. 2020).

A thorough revision of the S. camerunensis-gracilipes

group 1s in preparation. The size of the tympanum tn this

Species 1s approximately half the size of the eye.

Sclerophrys sp. aff. camerunensis 2

Fig. 6G—-H.

Area: Bechuchuu, Yalokole.

Season/survey: Wet (May 2018, Nov 2018), dry (Aug

2019).

Material: CSB:Herp:RNBK 195, 738, 739, 741-744;

IVB-H-CD 18165, 18282, 18310-18312.

Amphib. Reptile Conserv.

Comments: This species is similar to the previous one,

but seems to be rarer. It was found in swampy forests

near the Luo and Lopori rivers. There seems to be some

differences in the structure of verrucosity compared to

the previous species, which may indicate some ecological

differences. All our specimens have a more or less visible

light yellowish dorsal stripe. The size of the tympanum is

larger in this species, being approximately the size of the

eye. However, the margin of the tympanum (tympanic

annulus) is not very distinct.

Hyperoliidae

Afrixalus Laurent, 1944

The systematics of Central African Afrixalus has been

turbulent (e.g., Laurent 1982; Perret 1976b), and remains

unsettled. We found three species in Kokolopori, none

of which were ubiquitous or rare. Frogs of this genus

commonly have distinctly different color tones during

the day and night.

Afrixalus equatorialis (Laurent, 1941)

Fig. 7A-B.

Area: All.

Season/survey: Wet (May 2018, Nov 2018), dry (Aug

2019, Jul 2020).

Material: CSB:Herp:RNBK 016, 043, 095, 193, 272,

275, 280, 282, 296, 300, 342, 344, 346, 360, 392, 395,

437, 439, 457, 505, 509, 524, 537, 628, 654-657, 736,

757, 758, 804, 811; IVB-H-CD 18138, 18141, 18269,

18338, 18339; NMP-P6V 76073/1-4.

Comments: A_ relatively little-known A/frixalus

distributed only in rainforests of the northwestern to

central Congo Basin. This species was found along

streams in both disturbed and intact forest habitats. No

specimens were observed calling. One couple was found

in amplexus on the bank of the Lopori River. Dorsal

coloration is brown at night, but gray to whitish during

the day.

Afrixalus osorioi (Ferreira, 1906)

Fig. 7C.

Area: Yalokole, Yetee, Yotemankele.

Season/survey: Wet (May 2018), dry (Aug 2019).

Material: CSB:Herp:RNBK 093, 098, 135, 136, 150,

151, 153, 162, 165, 167, 174, 200, 338, 354, 381, 388,

398, 408, 435, 438, 454, 467, 508, 517, 593, 672, 674.

Comments: This species was described from west-

central Angola and is known to occur throughout much

of the Congo Basin, in Uganda, and in western Kenya.

The subspecies A. osorioi congicus (Laurent 1941)

was described from northern DRC, with the presumed

distribution in northern and eastern DRC (Laurent

1972). Later, Laurent (1982) noted that specimens of

A. osorioi from central Congo (Sankuru) were found to

be intermediate between the nominotypical subspecies

February 2022 | Volume 16 | Number 1 | e301

Amphibians of Kokolopori, central DR Congo

Fig. 7. Hyperoliids, Afrixalus, Congolius, and Cryptothylax. (A) Afrixalus equatorialis (male, nocturnal coloration). (B)

A. equatorialis (female, nocturnal coloration). (C) A. osorioi (male). (D) A. cf. quadrivittatus (amplectant pair). (E)

Congolius robustus (male). (F) C. robustus (female). (G) Cryptothylax greshoffii (male). (H) C. greshoffii (female).

Amphib. Reptile Conserv. February 2022 | Volume 16 | Number 1 | e301

Badjedjea et al.

and ssp. congicus and explained this pattern as a clinal

variation. Laurent agreed with other earlier authors

(e.g., Perret 1976b) to synonymize ssp. congicus with

the nominotypical subspecies. However, a detailed

investigation of the geographic morphological variation

and molecular phylogeography of the species may reveal a

more complex pattern in the future and possibly resurrect

the congicus taxon. This name could also be applicable

for the central Congolian population. In Kokolopori, A.

osorioi was found in ponds or along streams usually in

open habitats, but it was also recorded in disturbed flooded

forest together with A. cf. guadrivittatus. Most males were

found calling, hidden in higher herbaceous vegetation, and

some were found in amplexus. Based on a comparison of

three similar Congolian forest species (A. equatorialis, A.

leucostictus Laurent, 1950, and A. osorioz), Laurent (1982)

mentioned that A. /eucostictus can turn its coloration into

dark brown, almost without pattern, a condition which

rarely happens in A. osorioi. However, some of the

Kokolopori specimens of A. osorioi were dark brown,

with their dorsal pattern barely visible.

Afrixalus cf. quadrivittatus (Werner, 1908)

Fig. 7D.

Area: Yalokole, Yetee, Yotemankele.

Season/survey: Wet (May 2018, Nov 2018), dry (Aug

2019).

Material: CSB:Herp:RNBK 156, 172, 173, 175, 185,

196, 208, 231, 233, 402, 405, 424, 436, 510, 511, 658—

663, 673; IVB-H-CD 18094, 18135-18137, 18360-

18363, 18391-18395.

Comments: The striped Afrixalus from Central Africa

usually known as A. fulvovittatus (Cope, 1861) and/or

A. quadrivittatus seemingly represent more than two

species and form a species complex together with A.

dorsalis (Peters, 1875), which probably also represents

more than one species (Portik et al. 2019). Moreover,

the latter is also known to have a striped morph, which

further complicates morphological identifications

(Schigtz 1999). These striped Afrixalus have been known

under several names either as subspecies or full species

depending on the taxonomic authority as A. (fu/vovittatus)

brevipalmatus (Ahl, 1931), A. (fulvovittatus) leptosomus

(Peters, 1877), A. (fulvovittatus) quadrivittatus, or A.

“quadrivittatus” Pickersgill, 2007 (Amiet 2012; Amiet

and Goutte 2017; Frétey et al. 2011; Laurent 1982;

Pickersgill 2007; Schiotz 1999). The nomen /eptosomus

is now considered a synonym of A. dorsalis (Amiet

2012; Frétey et al. 2011) in line with the opinion of

earlier authors (Perret 1976b; Schiotz 1999). Afrixalus

fulvovittatus is the oldest available name for this frog

group. The type locality is in Liberia, and the species

is now believed to be distributed only in West Africa

(Channing and Rodel 2019). Afrixalus quadrivittatus has

the type locality in South Sudan and is believed to be

distributed in East Africa with unclear limits westward

in Central Africa (Channing and Rodel 2019). Frétey et

Amphib. Reptile Conserv.

al. (2011) hypothesized that this species is widespread

throughout Central Africa, similar to the hypothesis of

Laurent (1982; as subspecies A. f quadrivittatus). The

taxonomic situation is obviously complex (Portik et

al. 2019) and a molecular phylogeographic approach

is needed to resolve it. Afrixalus (fulvovittatus)

brevipalmatus is the available name applicable to the

Cameroonian and Equatorial Guinean populations (type

locality: Sangmelima, Cameroon; Amiet 2012; Amiet

and Goutte 2017; Sanchez-Vialas et al. 2020). For the

Kokolopori population we follow Laurent (1982), a

respected Congo herpetology expert, who hypothesized

that A. quadrivittatus is widespread from East Africa to

the Congo. However, in respect to the distance to the type

locality, we refer to this species as A. cf. guadrivittatus,

pending a thorough taxonomic revision. In Kokolopori,

we found this species mostly in open areas on higher

herbaceous vegetation, usually along streams, and

commonly together with A. osorioi.

Congolius Neéas, Badjedjea, and Gvozdik, 2021

This genus was recently established based on evidence

that “Hyperolius” robustus is phylogenetically placed

outside the genus Hyperolius (Neéas et al. 2021). The

Congo frog forms a common clade with West African

Morerella and Central African Cryptothylax. This new

and monotypic genus is probably endemic to the Central

Congolian forests.

Congolius robustus (Laurent, 1979)

Fig. 7E-F.

Area: All.

Season/survey: Wet (May 2018, Nov 2018), dry (Aug

2019, Jul 2020).

Material: CSB:Herp:RNBK 002, 009, 040, 050, 057,

061, 062, 067, 075, 081, 082, 096, 105, 111, 130, 137,

157, 215, 257, 414, 434, 445, 446, 466, 514, 519, 520,

532, 533, 543, 549, 550-552, 584, 606, 629, 679, 680,

698-700, 722, 730, 839-841; IVB-H-CD 18095, 18117-

18121, 18143, 18255, 18256, 18299, 18301-18305,

18364; NMP-P6V 76086/1-9, 76087/1-3.

Comments: This is a little-known species, but it 1s quite

common in Kokolopori. It is abundant in flooded primary

forests or along streams, and also in disturbed semi-open

habitats. It was commonly found in communities with

Hyperolius phantasticus and H. cf. cinnamomeoventris,

and/or H. cf. platyceps, especially in farmbush along

streams at forest edges. Most specimens were found

perched on vegetation 1.5—2 m high, but some specimens

were observed higher up on shrubs or trees. Most males

were found during calling activity, although they were

easily disturbed and stopped calling. The Robust Congo

Frog is sexually dichromatic with males in yellowish to

brown tones and usually dark marbling, whereas females

are reddish-brown with orange distal and ventral parts

of the limbs (especially tips of digits and webbings).

February 2022 | Volume 16 | Number 1 | e301

Amphibians of Kokolopori, central DR Congo

This species was already reported from Kokolopori by

Schiotz (2006).

Cryptothylax Laurent and Combaz, 1950

A large hyperoliid distributed in the western Congo

Basin, with one widespread species (C. greshoffii)

and one species of a smaller size (C. minutus) with

questionable validity (Schietz 1999) and known only

from its type locality near Lake Tumba, western-central

DRC (Laurent 1976b).

Cryptothylax greshoffi (Schilthuis, 1889)

Fig. 7G-H.

Area: Yalokole, Yetee.

Season/survey: Wet (May 2018, Nov 2018), dry (Aug

2019).

Material: CSB:Herp:RNBK 073, 146, 207, 256, 269,

248, 595; IVB-H-CD 18109, 18122-18127, 18232;

NMP-P6V 76077/1—10, 76078/1-3.

Comments: All observed adult specimens of Cryptothylax

were of large size, identified as C. greshoffii, and thus

did not fit the description of the enigmatic C. minutus

(Laurent 1976b). This species occurs in open areas

along streams or near swamps with rich high herbaceous

vegetation. Cryptothylax only rarely enters the margins

of forests. This sexually dichromatic species has

yellowish to brown males and whitish to pinkish females

with orange discs on the toes, which is a state of color

dichromatism similar to that of the genus Congolius.

Metamorphs of this species are relatively large and quite

tuberculous with beige to pinkish lateral stripes and a

brown dorsum, a pattern sometimes visible in adult males

but less conspicuous. We encountered this species only

relatively rarely, which is probably due to the presence of

predominantly forested habitats in Kokolopori.

HAyperolius Rapp, 1842

Our surveys recorded five species of this genus which

belong to two divergent clades (Portik et al. 2019): (1) H.

cf. cinnamomeoventris, H. cf. langi, H. cf. platyceps with

high color pattern variation but rather indistinct sexual

dichromatism; and (11) H. ocellatus, H. phantasticus

with a relatively stable color pattern but distinct sexual

dichromatism. Three of the five species are identified to

nominal species with some caveats as discussed below.

HAyperolius Clade 2 (sensu Portik et al. 2019)

HAyperolius cf. cinnamomeoventris Bocage, 1866

Fig. 8A—E.

Area: All.

Season/survey: Wet (May 2018, Nov 2018), dry (Aug

2019, Jul 2020).

Material: CSB:Herp:RNBK 091, 099, 109, 110, 113,

117, 128, 177, 206, 260, 274, 309, 334, 336, 348, 361,

Amphib. Reptile Conserv.

364, 387, 390, 412, 441, 447, 448, 450, 455, 460-465,

473, 512, 514-516, 604, 615, 665-667, 669-671, 759,

760, 843; IVB-H-CD 18092, 18093, 18107, 18108,

18128-18134, 18225, 18309, 18351, 18353-18359.

Comments: An abundant species in Kokolopori, found

in all habitat types but mostly in open areas along

streams, at forest edges, or in clearings. Phylogeographic

studies of this species suggested a more complex pattern

of genetic variation (Bell et al. 2015, 2017), and given

the relatively remote type locality in northern Angola,

the conspecificity of the central Congolian population

with the type-locality population is uncertain. The

Kokolopori population of this species possesses a high

level of color polymorphism, higher than presently

known for the species (cf. Amiet 2012; Channing

and Rédel 2019; Schigtz 1999). Generally, males are

yellowish to brown and females have a dominant green

color, which corresponds to the usual coloration (Amiet

2012; Schiotz 1999). However, one relatively atypical

feature is that light dorsolateral stripes are also present

in females (not only males), resembling H. veithi Schick,

Kielgast, R6dder, Muchai, Burger, and Lotters, 2010, but

less conspicuous (Fig. 8B; see also the photo of a pair in

amplexus in Ne¢as et al. 2021). This feature is a sign of

transition to secondary monochromatism, which is known

to occur in the H. cinnamomeoventris complex (Portik

et al. 2019). Another as yet undescribed color pattern is

an hourglass dorsal pattern present in some specimens

of both sexes (Fig. 8C—E). Schiotz (2006) observed both

color morphs of H. cf. cinnamomeoventris in Kokolopori

but identified them as “light” color morphs of H.

platyceps (Schigtz’s morphs C and D). In our opinion, H.

platyceps in Kokolopori always has a “dark” color morph

(see below). However, the dorsal brown coloration may

be relatively light under some conditions, which can

cause confusion in identification (cf. Figs. 8E and 9C).

Genetic testing could clarify whether the hourglass

morph (Schigtz’s morph C) represents a hybrid between

H. cf. cinnamomeoventris and H. cf. platyceps, as the

hourglass pattern is commonly present in H. platyceps

(Amiet 2012; Schiotz 1999). The taxonomic status of

certain taxa that are presumably conspecific or possibly

conspecific must be also verified, as some of these

nomina might be applicable for the central Congolian

populations of H. cf. cinnamomeoventris, particularly

H. ituriensis Laurent, 1943 described from Dyalasinda

near Lake Albert (Ituri Province), H. pol/i Laurent, 1943

described from Tshimbulu (Kasai-Central Province), and

H. (cinnamomeoventris) wittei Laurent, 1943 described

from Kulu near Mwanza, north of the Upemba National

Park (Haut-Lomami Province; Laurent 1957).

HAyperolius ct. langi Noble, 1924

Fig. 8F.

Area: Yalokole, Yetee.

Season/survey: Wet (May 2018), dry (Aug 2019).

Material: CSB:Herp:RNBK 159, 587-592, 610, 614.

February 2022 | Volume 16 | Number 1 | e301

Badjedjea et al.

Fig. 8. Hyperoliids, Hyperolius. (A) H. cf. cinnamomeoventris, striped morph (male). (B) H. cf. cinnamomeoventris, striped morph

(female). (C) H. cf. cinnamomeoventris, hourglass morph (male). (D) H. cf. cinnamomeoventris, hourglass morph (female); note

the species-specific red coloration of the inner thighs in both sexes in (C) and (D). (E) H. cf. cinnamomeoventris, hourglass morph

(male). (F) H. cf. /angi, striped morph, “forma albomarginata” (male). (G) H. phantasticus boulengeri (male, Phase J). (H)

H. phantasticus boulengeri (female).

Amphib. Reptile Conserv. February 2022 | Volume 16 | Number 1 | e301

Amphibians of Kokolopori, central DR Congo

Comments: This species seems to be relatively rare in

Kokolopori. Individuals were found calling and well

hidden in the dense herbaceous vegetation that overgrew

a small stream in farmbush at the edge of secondary

forest. This species has two distinct color morphs.

One has a camouflage-like pattern corresponding to

the “facies kuligae” (Laurent 1950), while the second

has light canthal and dorsolateral stripes extending

halfway down the body. We refer to the latter as the

“forma albomarginata’ (Fig. 8F) following the name

of the phenotypically corresponding nominal taxon

Hyperolius albomarginatus Laurent, 1940, synonymized

with H. /angi (Laurent 1950). There has been a long-

standing debate regarding the taxonomic distinction and

distributions of H. angi (type locality in Bas-Uele, NE

DRC) and H. kuligae (type locality on Mt. Cameroon),

e.g., Schiotz (1999). As the color pattern variation in H.

langi from northeastern DRC (unpub. data) basically

corresponds to the variation of H. kuligae in Cameroon

(cf. Amiet 2012), this suggests that the two species share

these phenotypic characteristics. We follow Kohler et

al. (2005), who studied advertisement calls, with the

opinion that the two species represent vicariants with

H. kuligae in western and H. /angi in eastern Central

Africa. The relatively deep genomic divergence also

supports the species status of both taxa (Portik et al.

2019). However, the exact distributional limits of the two

species are unknown. As the Kokolopori population is

relatively distant from the type locality (located north of

the Congo River), and also considering some differences

in coloration details, we refer to this population as

H. cf. langi. Interestingly, Bittencourt-Silva (2019)

reported a finding of the little-known H. major from

a forest patch in northwestern Zambia, supposedly

genetically close to H. angi and H. kuligae. However,

the photographed male differs in coloration from the

Kokolopori specimens. Individuals with the camouflage-

like pattern may be confused with H. cf. platyceps and

the hourglass specimens of H. cf. cinnamomeoventris,

as all three species have similar color pattern variations.

The photographed specimen of “H. kuligae” from Boteka

(western Equateur Province) reported by Schiotz (2006)

could be conspecific with H. cf. langi.

Hyperolius cf. platyceps (Boulenger, 1900)

Fig. 9C-E.

Area: All.

Season/survey: Wet (May 2018, Nov 2018), dry (Aug

2019, Jul 2020).

Material: CSB:Herp:RNBK 169, 170, 190, 192, 197,

214, 222, 236, 259, 261, 279, 291, 302, 318, 327, 333,

353, 365, 367, 376, 384-386, 413, 416, 433, 507, 651-

653, 668, 731-733, 805, 842; IVB-H-CD 18142, 18347-

18350, 18352, 18386, 18387.

Comments: A common, but often relatively well-hidden,

species 1n Kokolopori occurring in disturbed habitats or

at the edge of primary forest. This species is considered

Amphib. Reptile Conserv.

a Congolian faunal element (Amiet 2012), although we

have never encountered it in northeastern DRC. The

Species is widespread in the western and central Congo

Basin, Gabon, and northern Angola (Frétey et al. 2011;

Dewynter and Frétey 2019; Marques et al. 2018), and

possibly as far as southeastern DRC (Laurent 1952).

This species has been the source of much taxonomic

confusion in the past (Marques et al. 2018; Schiotz 1999)

and is not yet well understood. As mentioned above for

H. cf. cinnamomeoventris, Schietz (2006) described

four color morphs from Kokolopori: two dark and two

light. However, based on our close observations in the

field and in the laboratory, we are of the opinion that the

“light morphs” represent H. cf. cinnamomeoventris (with

the possibility of a hybrid origin of the light hourglass

morph, Schietz’s morph C, which remains to be

genetically tested; Fig. 8C—E). All the specimens which

we identified as H. cf. platyceps belonged to the two dark

morphs, one with the hourglass dorsal pattern (Fig. 9C)

and the second one a rarer striped morph corresponding

to the “forma pleurotaenia” (cf. Amiet 2012; Fig. 9D).

As noted by Schigtz (2006), individuals from Kokolopori

have some peculiarities in coloration that are not known

in the western populations. In particular, conspicuous

white and/or yellow spots are present on the flanks and

part of the ventrum in both sexes (spots of both colors

may be combined; see Fig. 9E for ventral view).

Several subspecies were described, including two

from southeastern DRC: H. platyceps lomamiensis

Laurent, 1943 (type locality in W Upemba NP, Haut-

Lomami) and H. p. olbrechtsi Laurent, 1952 (type

locality W of Kalemie, Tanganika). Hyperolius langi

and H. major Laurent, 1957 (type locality in E Upemba

NP, Haut-Katanga) were also treated as subspecies of

H. platyceps at one time (e.g., Laurent 1952, 1957).

However, Schigtz (1999) stated: “Much confusion has

surrounded this species, partly because of its variation,

partly because several quite different species have been

treated as subspecies of H. platyceps. Amiet (1978)

[Note: Amiet 1979 “1978”] has clarified the matter and

we follow him in regarding the forms in the “platyceps-

complex” as full species (platyceps, langi, major), but

did not address the taxa /Jomamiensis and olbrechtsi. Both

are presently treated as synonyms (however, they may

be valid subspecies) of H. platyceps (Frost 2021). From

the above, it is clear that a thorough revision 1s needed

to clarify the status of these taxa and their relationships

to H. langi and H. major. It is also necessary to verify

whether H. p. Jomamiensis is an older available name for

H. major as the type localities of the two taxa are only

about 70 km apart. Similarly, some other little-known

taxa may be related to H. platyceps (cf. Laurent 1941,

1943, 1952, 1957), in particular H. atrigularis Laurent,

1941 described from highlands of the Marungu Plateau

(Tanganika Province), H. polli Laurent, 1943 known

from the type locality in Kasai-Central and adjacent

northeastern Angola (Marques et al. 2018), and possibly

February 2022 | Volume 16 | Number 1 | e301

Badjedjea et al.

Fig. 9. Hyperoliids, Hyperolius and Hylambates. (A) Hyperolius phantasticus boulengeri (amplectant pair, both sexes in Phase F).

(B) H. phantasticus boulengeri under UV light (male, Phase J). (C) H. cf. platyceps, hourglass morph (male). (D) H. cf. platyceps,

striped morph, “forma pleurotaenia” (male). (E) H. cf. platyceps, hourglass morph, ventral view (female). (F) H. ocellatus

purpurescens (female). (G) Hylambates verrucosus (male). (H) H. verrucosus, ventral view (same specimen as in G).

Amphib. Reptile Conserv. 55 February 2022 | Volume 16 | Number 1 | e301

Amphibians of Kokolopori, central DR Congo

Hyperolius kibarae Laurent, 1957 described from the

eastern Upemba NP (Haut-Katanga Province).

HAyperolius Clade 1 (sensu Portik et al. 2019)

HAyperolius ocellatus Giinther, 1858

Hyperolius ocellatus purpurescens Laurent, 1943

Fig. 9F.

Area: Yetee.

Season/survey: Dry (Aug 2019).

Material: CSB:Herp:RNBK 611, 612, 694, 695-697.

Comments: This species was recorded only once near

Yalokenge village in pristine forest, in a partially open

swampy zone at the edge of a stream. Five males were

found sitting on vegetation, and not calling. The only

female was discovered well hidden in vegetation, far

from the place where the males aggregated. Schiotz

(2006) also recorded “only few specimens” in small

forest swamps in Kokolopori, and described their

coloration. This is a sexually dichromatic species (Portik

et al. 2019) with green males that have a silvery white

triangle on the snout and white dorsolateral stripes. The

female (Fig. 9F) has the color pattern corresponding to

the subspecies H. o. purpurescens originally described

from the southeast of Kisangani (Laurent 1943) [type

locality of the nominotypical subspecies was restricted

to Bioko Island, Equatorial Guinea]. A phylogeographic

study suggested a complex pattern of genetic variation

with the eastern Congolian populations differentiated to

some level (samples from Kokolopori were not included;

Bell et al. 2017). Hyperolius hildebrandti Ahl, 1931

(type locality “Kamerun;” Ahl 1931) may be an older

available name for populations from the Congolian

forests. Whether this taxon deserves full species status

remains to be evaluated.

HAyperolius phantasticus (Boulenger, 1899)

Hyperolius phantasticus boulengeri Laurent, 1943 (new

status)

Figs. 8G—H, 9A-B.

Area: Yalokole, Yetee, Yotemankele.

Season/survey: Wet (May 2018, Nov 2018), dry (Aug

2019).

Material: CSB:Herp:RNBK 001, 003, 010, 013, 021,

033, 164, 168, 176, 187, 189, 199, 202, 205, 213, 217,

223,238; .301,. 303-305, 307,308, 330, 337,356, 357,

359, 363, 368-370, 374, 382, 383, 393, 394, 399, 403,

406, 409, 410, 431, 440, 449, 459, 468-472, 474, 475,

479, 480, 571, 579, 580, 605, 616, 701; TVB-H-CD

18087-18091, 18096-18099, 18110-18116, 18162-

18164, 18191, 18192, 18202, 18203, 18294, 18342-

18346, 18492, 18493, 18508, 18518, 18519, 18523,

18526, 18528.

Comments: Hyperolius phantasticus is distributed from

southern Cameroon to western Congo/DRC and to central

DRC (Channing and Rodel 2019), and was described from

the present-day continental Equatorial Guinea (Sanchez-

Amphib. Reptile Conserv.

Vialas et al. 2020). It is one of the most abundant anuran

species in Kokolopori, usually present along streams

and rivers in open or semi-open areas, perching on high

herbaceous vegetation and shrubs. Many pairs were

found in amplexus during the wet seasons, while no

amplectant pairs were found during the dry season. In

this region, this species occurs in two color phases as

already described by Schigtz (2006): Phase J (Fig. 8G;

juveniles/males; Schiotz 1967) with the predominately

light, translucent green dorsal color, unspotted or

sprinkled with tiny dark spots, canthal stripes (light or

dark) in most specimens, in some individuals continuing

behind the eye halfway down the body, venter mostly

yellow-green with some blue patches; and Phase F (Figs.

8H, 9A; females/female-like males) with the maroon to

beige dorsal color, sometimes with small yellow dots,

ventral side mostly ink-black suffused with bluish and

pinkish tones in dispersed light patches (sometimes

concentrated in the center while circum-marginal areas

are black). The inflated throat of males is mostly of the

blue-green color in both color phases, with the gular disc

yellow in Phase J and showing the dorsal maroon color

in Phase F. The coloration differs significantly from the

holotype of H. phantasticus (female; Boulenger 1899)

and the populations of Cameroon and northern Gabon

(cf. Amiet 2012; Amiet and Goutte 2017; Dewynter and

Frétey 2019), which are characterized by yellow dorsal

coloration without spots, only with dark canthal stripes,

and the ventral side is ink-black only in the posterior

parts (Amiet 2012; Boulenger 1899). These geographic

differences in coloration have already been discussed

(Amiet 2012; Kohler et al. 2005). We therefore propose

to resurrect Hyperolius boulengeri Laurent, 1943 [type

locality “Flandria (Tshuapa),” DRC; Laurent 1943] at

the subspecies level as H. phantasticus boulengeri (new

status) to account for geographic variation with the

consistent color differentiation and genetic divergence

(unpub. data; genetic data to be published elsewhere)

of the central Congolian populations. It is possible that

this taxon occurs as far west as southern Gabon and

southwestern Republic of the Congo as the coloration

of H. phantasticus in these regions (cf. Jongsma et al.

2017; Largen and Dowsett-Lemaire 1991) roughly

corresponds to individuals from central DRC. However,

a phylogeographic study is needed to clarify the

distribution range of this taxon and whether it deserves

full species status. We have observed that this species is

preyed upon by fishing spiders (Ni/us, Pisauridae), with

a male of Phase F found as a prey item in May 2018

(Badjedjea et al. 2019), and a male of Phase J as a second

case in the dry season of August 2019. Interestingly, we

observed a certain level of the fluorescent emissions in

adult males of Phase J (Fig. 9B) upon illumination with

a UV light (A, 395-400 nm), which agrees with the

hypothesis of the existence of fluorescence in anurans

with greenish-translucent skin (Taboada et al. 2017a,b).

Fluorescence was not observed in males of Phase F.

February 2022 | Volume 16 | Number 1 | e301

Badjedjea et al.

HAylambates Duméril, 1853

This genus of the subfamily Kassininae (while the

previous genera are representatives of the subfamily

Hyperoliinae) was known as Philyctimantis from the

1950s until recently, when Dubois et al. (2021) pointed

out the nomenclatural priority of the nomen Hy/ambates.

These treefrogs have only rarely been encountered in

Kokolopori.

Hylambates verrucosus Boulenger, 1912

Fig. 9G—H.

Area: Bechuchuu, Yalokole.

Season/survey: Wet (Nov 2018), dry (July 2020).

Material: CSB:Herp:RNBK 810, 819, 820, 827; NMP-

P6V 76085/1-2.

Comments: Individuals of this species were found

in forest swamps dominated by plants of the genus

Lasimorpha (formerly known as Cyrtosperma). This

semiaquatic plant is spiny and may provide some

protection from the predators of these frogs. Three anuran

species were abundant in this swampy environment,

Aubria masako and Chiromantis cf. rufescens in addition

to H. verrucosus. In our field experience in central and

northeastern DRC, Hylambates is often syntopic with

Chiromantis.

Phrynobatrachidae

Phrynobatrachus Giinther, 1862

The systematics of Congolian Phrynobatrachus is poorly

known. We have identified three morphologically similar

species in our Kokolopori material, all belonging to the P.

auritus species complex. The identification of these frogs

is complicated by the high level of color polymorphism

and variation in their color patterns. For example, light

stripes may be missing or may occur medially or laterally,

sometimes bright spots (e.g., green) are distributed in

different places, etc. This species complex is currently in

taxonomic revision (V. GvoZdik et al., in prep.).

Phrynobatrachus cf. auritus Boulenger, 1900

Fig. 10C.

Area: Yalokole.

Season/survey: Wet (May 2018).

Material: CSB: Herp:RNBK 066, 101.

Comments: The type locality of this species is far from

the study site, in Equatorial Guinea (Sanchez-Vialas et

al. 2020), thus this identification still needs to be tested

by molecular methods. Individuals corresponding to

this species were found only near the Eho stream, in the

neighborhood of the Bandjangi swamp (used by local

people to macerate cassava) at the edge of disturbed

forest and farmbush, near Yalokole village. This record

could indicate a higher level of tolerance to disturbed

habitats, as the other two species were found only in

Amphib. Reptile Conserv.

primary forest. The general coloration of this species

appears to be rather lighter, with a mostly pale, whitish

venter.

Phrynobatrachus cf. giorgti De Witte, 1921

Fig. 10B.

Area: Bechuchuu, Yalokole.

Season/survey: Wet (May 2018, Nov 2018), dry (Aug

2019, Jul 2020).

Material: CSB:Herp:RNBK 254, 718, 737, 761, 762,

855, 856, 857, 858, 859, 860, 861; IVB-H-CD 18190,

18237, 18238, 18283, 18284, 18297, 18298, 18300;

possible hybrids: IVB-H-CD 18186, 18188.

Comments: Individuals of this species morphologically

correspond to the type material of P. giorgii. However,

the type locality (Yambata) 1s north of the Congo River

and therefore the identification must still be tested

genetically. This species was found at several sites,

always in primary forest and usually near small streams

or swampy places in flooded forest. Some males were

found calling from moist leaf-litter. Depending on the

physiological conditions, this species may have smooth

skin, but quite often has slightly warty skin. It is very

variable in color tones (and patterns), but often has a

lighter background (reddish to brown) suffused with

black. One specimen (adult female) was very dark,

almost black, on the dorsum and darkly marbled on

a white venter. Adult males have black throats. This

relatively small-sized species was found in syntopy with

another Phrynobatrachus, a slightly larger and generally

lighter species resembling P. auritus (P. sp. aff. auritus,

see below). As some individuals had intergrading traits,

it 1s possible that the two species hybridize, but this needs

to be confirmed genetically.

Phrynobatrachus sp. aff. auritus

Fig. 10A.

Area: Yalokole.

Season/survey: Wet (May 2018, Nov 2018).

Material: CSB:Herp:RNBK 107; IVB-H-CD 18185,

18187, 18189, 18239.

Comments: Only a few specimens of this presumably

new species resembling P. auritus were recorded. All

were found in primary forest, most of them syntopically

with P. cf. giorgii in a swampy place surrounded by

flooded forest on the bank of the Luo River. One

specimen was found nearby, near the Bikongo stream in

terra firma forest. The general coloration of this species

seems to be lighter than in P. cf. giorgii, the venter is

white, and the throat in adult males is gray or brownish

but not deep black.

Pipidae

Hymenochirus Boulenger, 1896

This genus is well known from aquaria (Kunz 2007; Rabb

February 2022 | Volume 16 | Number 1 | e301

Amphibians of Kokolopori, central DR Congo

tri

id tad Ue)

és

Fig. 10. Phrynobatrachids (Phrynobatrachus), ptychadenids (Ptychadena), and a pyxicephalid (Aubria). (A) Phrynobatrachus

sp. aff. auritus (male). (B) Ph. cf. giorgii (male). (C) Ph. cf. auritus (male). (D) Ptychadena aequiplicata (female). (E) Pt.

christyi (male), extralimital specimen from Mosite (0.956°N, 23.560°E). (F) Pt. perreti (male). (G) Pt. sp. aff. mascareniensis

(male). (H) Aubria masako (male).

Amphib. Reptile Conserv. February 2022 | Volume 16 | Number 1 | e301

Badjedjea et al.

and Rabb 1963), but wild populations are poorly known.

Similarly, knowledge of the taxonomy of this genus is

poor, with only four species currently listed (Frost 2021).

The distribution of this genus is concentrated around the

forested zone of Central Africa, from southern Nigeria to

eastern DRC, and possibly westernmost Uganda.

Aymenochirus cf. boettgeri (Tornier, 1896)

Fig. 11A.

Area: Bechuchuu, Yalokole.

Season/survey: Wet (Nov 2018), dry (July 2020).

Material: CSB:Herp:RNBK 831; IVB-H-CD 18216-—

18220, 18308, 18397-18401, 18402-18406, 18410-

18424, 18456-18465, 18480-18488, 18529, 18551.

Comments: This species is common in Kokolopori,

usually found in syntopy with Xenopus pygmaeus

in stagnant water in forests, typically in pools at the

periphery of flooded forest. The general appearance

resembles the type of H. boettgeri (ZMB 11521), which

was described from the Ituri Forest, northeastern DRC.

Given the relatively distant type locality in a different

ecoregion, we assume that this population may represent

a distinct taxon.

Hymenochirus cf. boulengeri De Witte, 1930

Fig. 11B.

Area: Yalokole.

Season/survey: Wet (Nov 2018).

Material: [VB-H-CD 18376, 18396, 18578-18580.

Comments: Another species of Hymenochirus, but

apparently much rarer, was found in shallow slow-

flowing parts of a stream located at the edge of forest

and farmbush. It was found in syntopy with the previous

species, but this species was never found deeper in the

forest. Similar to H. cf. boettgeri, the overall morphology

of the Kokolopori individuals corresponds to the type

material of H. boulengeri described from the forest-

savanna mosaic in northern DRC (De Witte 1930).

However, with respect to the relatively distant type

locality, we refrain from making a definite identification

of the species until further material has been examined.

Xenopus Wagler, 1827

Based on current knowledge, this species-rich genus of

pipid frogs appears to be surprisingly represented by

only a single species in the central Congo. However, the

morphology of species of the X. amieti group, to which

this single species belongs, is relatively uniform (Evans

et al. 2015, 2019) so other overlooked species may be

present.

Xenopus pygmaeus Loumont, 1986

Fig. 11C—D.

Area: Bechuchuu, Yalokole.

Season/survey: Wet (May 2018, Nov 2018), dry (Jul

2020).

Amphib. Reptile Conserv.

Material: CSB:Herp:RNBK 118, 160, 163, 219, 224,

265, 316, 319-321, 325, 340, 341, 351, 352, 358, 371,

372, 379, 401, 422, 821, 828-830, 845-853; IVB-H-CD

18204—18215, 18314-18322, 18407-18409, 18479.

Comments: This species is common in stagnant water,

usually in swampy areas in forests. It is harvested as

food to a limited extent, mostly by children and young

people. We did not find tadpoles in the wild, but this

species has been bred in captivity and a tadpole is shown

in Fig. 11D. The general morphology corresponds to the

typical Xenopus tadpole morphology (Channing et al.

2012; Vigny 1979). Tadpoles reached a total length of

49 mm (stage 42; Gosner 1960), with the tail (33 mm)

approximately twice as long as the body (16 mm), and

metamorphosed in 6—8 weeks.

Ptychadenidae

Ptychadena Boulenger, 1917

Representatives of this genus are not very abundant in

Kokolopori. Four species were recorded, mostly in and

around villages in open habitats and/or farmbush and

secondary forest. Only P. aequiplicata was mostly found

in primary forest. Records of all four species represent

significant geographic range extensions. Taxonomically,

there is a great deal of confusion around the Congolian

Species as discussed below.

Ptychadena aequiplicata (Werner, 1898)

Fig. 10D.

Area: Bechuchuu, Yalokole.

Season/survey: Wet (May 2018, Nov 2018), dry (Aug

2019, Jul 2020).

Material: CSB:Herp:RNBK 123, 735, 752, 803, 806,

833, 834; IVB-H-CD 18293.

Comments: Piychadena aequiplicata is characterized by

extended dark (blackish) webbing and numerous shorter

irregular dorsal glandular ridges/folds (Frétey et al.

2011; Channing and Rodel 2019). This species has been

reported from West Africa (with taxonomic uncertainty,

possibly representing an undescribed species) to western

Central Africa, and only from the westernmost margin of

DRC (Channing and Rodel 2019; IUCN SSC Amphibian

Specialist Group 2019). However, P. aequiplicata is

relatively widespread in western to central DRC (unpub.

data), and our records from Kokolopori represent a

substantial geographic range extension eastward to the

Central Congolian forests. We have also recorded this

species further east, around Kisangani. It is possible

that this species is even more widespread, distributed

throughout the Congo Basin and entering Uganda.

This is indicated by the published record from “Tturi:

Madié” [= Medje, Haut-Uélé] (Boulenger 1919), if the

identification was correct, and by photographs showing

“P. christy’ (Channing and Rodel 2019: p. 341) from

Budongo, Uganda (T.M. Doherty-Bone, pers. comm.),

February 2022 | Volume 16 | Number 1 | e301

Amphibians of Kokolopori, central DR Congo

Fig. 11. Pipids (Hymenochirus, Xenopus), ranids (Amnirana), and a rhacophorid (Chiromantis). (A) Hymenochirus cf. boettgeri

(male). (B) H. cf. boulengeri (male). (C) Xenopus pygmaeus (female). (D)_X. pygmaeus, tadpole. (E) Amnirana cf. albolabris

(male). (F) Amnirana cf. albolabris, dark morph with contrasting color pattern (male). (G) A. /epus (male). (H) Chiromantis cf.

rufescens (male).

Amphib. Reptile Conserv. 60 February 2022 | Volume 16 | Number 1 | e301

Badjedjea et al.

but morphologically corresponding to P. aequiplicata.

This hypothesis needs to be further tested, but it is

already evident that the species is distributed at least to

central and northeastern DRC. Pitychadena aequiplicata

is probably the most strongly associated with forest

habitats of all the representatives of this genus. It appears

to be relatively rare in Kokolopori, and it was only found

in primary forest.

Ptychadena christyi (Boulenger, 1919)

Fig. 10E.

Area: Bechuchuu, Yalokole, Yetee.

Season/survey: Wet (Nov 2018), dry (Aug 2019).

Material: CSB:Herp:RNBK 636, 637, 664, 720; IVB-

H-CD 18306.

Comments: This species is very poorly known and

is somewhat enigmatic because some other species

of Ptychadena have been misidentified as P. christyi.

Here, we probably show the first published photograph

of this species, as the photographed frog published by

Noble (1924) does not conform to the morphology of the

types of P. christyi. The frog in that figure (Noble 1924)

resembles P. /ongirostris (Peters, 1870) from West Africa,

which can indicate either a substantial range extension

of this species or that a similar species has remained

overlooked in northeastern DRC. Ptychadena christyi

was described from “Madié (Itur1)” [= Medje, Haut-Uéelé]

based on three males and one female (Boulenger 1919).

One male syntype is located in BMNH (1947.2.2.59;

formerly 1919.8.16.20); and two male and one female

syntypes, together with one juvenile also marked as a

type (although not listed in the original publication), are

stored in RMCA (B.324—326: adults; B.311 juvenile).

This species is characterized by its pronounced

dorsolateral glandular ridges/folds and usually smooth

dorsum, in contrast to most other Ptychadena species

which have numerous distinct longitudinal dorsal folds.

Some individuals bear additional short irregular dorsal

and lateral folds, which are less conspicuous than the

two dorsolateral ones. This is probably why P. christyi

has sometimes been considered a close relative of, if not

conspecific to, P. aequiplicata (Noble 1924). However,

these two species are clearly different at first glance (cf.

Figs. 1OD-E). Zimkus and Larson (2013) suggested that

P. christyi may be conspecific with P. perreti. However,

this assumption was based on P. perreti from Uganda

which was misidentified as “P. christyi’ (per our own

review of the data). Based on the material stored in

RMCA and our own field observations, P. christyi is

known only from the forested regions of northeastern

DRC. The Kokolopori material thus demonstrates a

geographic range extension to the south. It is not clear

if the species has been reliably recorded from Uganda,

despite reports of it from the country. Based on the

available data, it seems that many if not all reports of

P. christyi from Uganda were misidentifications (see

above and accounts for P. aequiplicata and P. perreti).

Amphib. Reptile Conserv.

In Kokolopori, we found P. christyi in puddles or pools

on muddy roads formed after heavy rains, where they lay

eggs. If the pools were near a bush, the frogs jumped

out of the pools to hide in the bushes when disturbed.

Some individuals were found in the primary forest at

Bechuchuu, in a pirogue (dugout canoe) filled with water.

Ptychadena perreti Guibé and Lamotte, 1958

Fig. 10F.

Area: Yalokole, Yetee.

Season/survey: Wet (May 2018, Nov 2018).

Material: CSB:Herp:RNBK 008, 029, 032, 051, 059,

115, 127, 133, 134, 145, 148, 161, 180, 228, 396, 444,

526, 640, 646; [VB-H-CD 18069, 18166, 18167.

Comments: Ptychadena perreti was described from

southern Cameroon (Guibé and Lamotte 1958), and is

believed to be distributed in western Central Africa and

north of the Congo River to northeastern DRC (Channing

and Rédel 2019; IUCN SSC Amphibian Specialist Group

2020). Our Kokolopori material provides evidence for a

geographic range extension into the Central Congolian

forests. Based on the publicly available genetic data

(our own review of data), “P christyi’ from Uganda

(Bundibugyo; GenBank No. GQ183595)_ represents

misidentified P. perreti, proving evidence for the presence

of this species also in western Uganda. Similarly, “P.

bibroni” (or P. aff. bibroni) from Gabon (GenBank

AY517604; Vences et al. 2004) and “P. cf. aequiplicata”

from Cameroon (Mt. Kupe; GenBank KX671789; Portik

et al. 2016) both represent misidentified P. perreti,

as well as “Ptychadena sp. B” from the Republic of

the Congo (Deichmann et al. 2017). In the past, this

species was probably at least partially misidentified as

“P. oxyrhynchus” (originally in the genus Rana) in the

Congo, as the photograph published by Noble (1924)

with this name most probably depicts P. perreti. All these

past misidentifications have caused further confusion in

the identification of this species. Ptychadena perreti 1s

a widespread inhabitant of the forested zone of Central

Africa. However, it is usually found in degraded forests,

shrubs, and farmbush. In Kokolopori, we found it in

swampy places along streams, usually in disturbed forest.

Ptychadena sp. aff. mascareniensis

Fig. 10G.

Area: Yalokole, Yotemankele.

Season/survey: Wet (May 2018, Nov 2018), dry (Aug

2019).

Material: CSB:Herp:RNBK 069, 080, 120, 149, 184,

531, 641, 642, 643, 677, 678; IVB-H-CD 18221, 18226-

18229, 18231, 18233-18235, 18272-18274.

Comments: This species was usually found directly in

villages, or nearby in periodical muddy pools formed

after heavy rains. This species does not enter pristine

forest, but can be found along streams in swampy places

at forest edges. It may be syntopic with other Ptychadena

species, e.g., P. perreti. The P. mascareniensis species

February 2022 | Volume 16 | Number 1 | e301

Amphibians of Kokolopori, central DR Congo

complex is distributed throughout sub-Saharan Africa,

usually in open habitats, including human settlements and

agricultural sites (R6del 2000). It enters forested regions

along roads. Twelve evolutionary lineages were identified

in a recent study (Zimkus et al. 2017), with only three of

them named. In addition to P. mascareniensis comprising

at least three evolutionary lineages from Réunion,

Mauritius, Seychelles, and Madagascar, P. nilotica is

known from southern to northeastern Africa, including

eastern Africa, and P. newtoni is endemic to Sdo Tomé

Island in the Gulf of Guinea. The remaining lineages have

remained unresolved taxonomically (usually named P.

cf. mascareniensis or P. “mascareniensis”, Frost 2021),

treated as operational taxonomic units (OTUs). In DRC,

P. nilotica and three OTUs (6, 8, 9) have been recorded,

of which two have been found in forested lowlands

(OTU 6 and OTU 8). From available data, it seems that

OTU 8 is confined to the central and eastern Congolian

forests (Zimkus et al. 2017). There is probably no name

available for OTU 8. If it proves to be distinct, 1t needs to

be described formally as a new species.

Pyxicephalidae

Aubria Boulenger, 1917

This genus contains two species, but only one of them is

distributed in the Congo Basin.

Aubria masako Ohler and Kazadi, 1990

Fig. 10H.

Area: Bechuchuu, Yalokole.

Season/survey: Wet (May 2018, Nov 2018), dry (Jul

2020).

Material: CSB:Herp:RNBK 253, 818, 822-826, 837;

IVB-H-CD 18170, 18230, 18365, 18366.

Comments: This species was described from the Masako

Forest Reserve, near Kisangani, DRC (Ohler and Kazadi

1990). It seems to be confined to swampy rainforests of

the Congo Basin (Channing and Rodel 2019). There is

an ontogenetic shift in the coloration of the venter, with

juveniles having a black venter with white spots, later

changing into gray with yellow spots, and finally to a

mostly yellow venter. This robust frog occupies swamps,

often in flooded forests, but sometimes in places where

cassava is macerated by villagers. The local people of

Kokolopori occasionally collect these frogs as bushmeat,

especially children and women, but the collection is not

intensive. Males were well hidden when calling, sitting

in burrows under tree roots or even underwater. Their

advertisement call resembles the sound of a pig.

Ranidae

Amnirana Dubois, 1992

Two species of this genus were found in Kokolopori, one

Amphib. Reptile Conserv.

present mostly in disturbed habitats, while the second

was found only in forest.

Amnirana cf. albolabris (Hallowell, 1856)

Fig. 11E-F.

Area: All.

Season/survey: Wet (May 2018, Nov 2018), dry (Aug

2019, Jul 2020).

Material: CSB:Herp:RNBK 004—006, 011, 014, 015,

017, 024, 026, 030, 039, 042, 046-049, 053, 055, 060,

077, 079, 090, 097, 100, 102, 104, 106, 112, 166, 191,

425, 427-430, 476-478, 502-504, 525, 529, 553-560,

565, 566, 576, 607, 608, 650, 685, 708-710, 723-726,

745-747, 753-755, 835, 836, 838, 862-865; IVB-H-CD

18066-18068, 18083, 18084, 18171-18175, 18390.

Comments: This species belongs to the taxonomically

unresolved A. albolabris species complex (Jongsma et al.

2018), distributed mostly in Central Africa from Nigeria

to Uganda (Channing and Rodel 2019). The Kokolopori

population probably belongs to the central Congolian

evolutionary lineage. Some individuals have contrasting

coloration with a dark dorsum, bright lips and spots on

the flanks, and a distinctly marbled venter (Fig. 11F), a

color pattern not known to us from other parts of Central

Africa. However, most individuals are lighter and not

as contrasting (Fig. 11E). This species was commonly

found in most habitats, but especially in disturbed places

near human settlements and along streams. This species

does not seem to enter deeper primary forests. The

tadpoles are contrastingly colored with black spots on a

red background. The skin glands of tadpoles produce a

poisonous secretion, making them unpalatable to most

vertebrate predators (Channing et al. 2012).

Amnnirana lepus (Andersson, 1903)

Fig. 11G.

Area: Yalokole, Yetee, Yotemankele.

Season/survey: Wet (May 2018, Nov 2018), dry (Aug

2019).

Material: CSB:Herp:RNBK 131, 155, 179, 209, 210,

230, 419, 561-564, 567-569, 575, 609, 682-684, 686—

689, 692, 693; IVB-H-CD 18052, 18064, 18065.

Comments: This species is known from Cameroon to

DRC, but has also recently been recorded in northern

Angola (Jongsma et al. 2018; Ernst et al. 2020).

Compared to A. cf. albolabris, it has been encountered

relatively rarely in Kokolopori. It is more confined to

forested habitats than the previous species. Females are

much larger than males.

Rhacophoridae

Chiromantis Peters, 1854

The only species of this genus, C. rufescens, is known

from Central Africa. However, a recent phylogeographic

study has shown that the north-central and eastern

February 2022 | Volume 16 | Number 1 | e301

Badjedjea et al.

Congolian population is relatively divergent (the

estimated origin 1s dated to the Pliocene/Pleistocene

boundary) and may represent a distinct species (Leaché

et al. 2019). Representatives of this genus deposit eggs

into foam nests attached usually to branches above water

(Channing and Rédel 2019). The foam is produced from

a secretion during oviposition by beating the secretion by

both females and males, sometimes including multiple

males.

Chiromantis cf. rufescens (Ginther, 1869)

Fig. 11H.

Area: Bechuchuu, Yalokole, Yotemankele.

Season/survey: Wet (May 2018, Nov 2018), dry (Aug

2019, Jul 2020).

Material: CSB:Herp:RNBK 218, 235, 255, 283, 538,

539, 814-817; IVB-H-CD 18236.

Comments: A relatively rarely encountered species in

Kokolopori. Foam nests with adults sitting nearby were

observed on vegetation in secondary forest during both

the wet and dry seasons (May 2018, Jul 2020). Some foam

nests were found almost on the ground near a swamp. It

is not known how far into the Central Congolian forests

this species is distributed, however some specimens in

the RMCA are also from the more western province of

Equateur.

Discussion

The fauna of the Central Congolian Lowland Forests

ecoregion is one of the least-known faunas of terrestrial

ecosystems in the world (Burgess et al. 2004). For

example, a long-standing mystery is the alleged absence

of caecilians (Nussbaum and Pfrender 1998). Whether

caecilians are present remains unclear, but the amphibians

of the Central Congo have never been studied in detail.

The main reason for the lack of research is the ongoing

recent socio-political instability, including civil wars,

and the limited accessibility which persists to the present

day (Anthony et al. 2015; Greenbaum 2017).

Here we present the results of the first targeted,

relatively long-term amphibian survey of the Central

Congolian forests carried out in Kokolopori. The species

identifications were based on careful examinations of

new and available comparative material, including types.

The surveys for amphibians used a variety of techniques,

and the occurrence of caecilians was surveyed, but not

confirmed, among local residents based on photographs

of various African caecilians. During four field sessions

lasting a total of 48 days in both the wet and dry

seasons, 37 anuran species were detected (Figs. 4-11,

Table 2). The amphibian fauna of Kokolopori is mainly

represented by forest species, which are more widely

distributed in the lowland rainforests of Central Africa.

However, precise species identification was not always

possible and indicates the likely presence of hitherto

undescribed species, and thus some degree of endemism.

Amphib. Reptile Conserv.

No caecilians were found, nor did the local people claim

to know of them. The local people mostly confused

them with blind snakes or limbless skinks, which may

indeed indicate the absence of caecilians in the Central

Congolian forests.

The species richness statistics estimated a total

of 37-41 species (Fig. 3, Table 1), suggesting that

our survey could be nearly complete. However, we

assume that some rare or hard-to-detect species with

cryptic ecology have probably been overlooked, e.g.,

Schigtz (2006) recorded one species not found in our

surveys, H. brachiofasciatus (see below). Interestingly,

approximately 75-80% of the species were recorded

during each of the two-week surveys, suggesting that

even just a single two-week intensive survey may have

good potential for amphibian inventories in Afrotropical

forests. On the other hand, the two-week surveys

provided relatively poor data for statistical estimates of

species richness, with large standard deviations and wide

confidence intervals. However, the means converged to

similar values, especially in the wet season analyses. The

dry season analysis estimated a slightly lower number

of species because the number of species observed was

also slightly lower (27 species observed in the dry season

compared to 30-31 during the wet-season sessions). The

difference between the number of species observed in

the dry and wet seasons is not large, but bear in mind

that the dry season in Kokolopori is also relatively wet

with occasional rains. In regions with a highly seasonal

climate, the number of species observed and/or estimated

in the dry season may be much lower than in the wet

season. The relatively low number of species observed

and estimated compared to some other Central African

regions, such as the coastal forests around the southern

Cameroon Volcanic Line (Herrmann et al. 2005; Nneji

et al. 2021), can probably be attributed to the climatic

history of the central Congo, which was subjected to

severe droughts during the cyclic glacial periods of the

Pleistocene (Maley 1987, 1991, 1996; Plana 2004). On

the other hand, some degree of endemism must result

from the persistence of riverine forest refugia in the

region (Colyn et al. 1991; Leaché et al. 2019; Maley

1996; Plana 2004).

Most species recorded in Kokolopori were typical

forest specialists, although habitat generalists also

occurred to some extent. Among the most frequently

encountered species were the scansorial Leptopelis

ocellatus, _Congolius robustus, and Hyperolius

phantasticus typically found on the forest edges or in

forests, and Amnirana cf. albolabris frequently found

in degraded habitats. The most abundant species on

average was H. phantasticus (typically along streams at

the forest edges), but its abundance was not so evident

in the dry season. During the wet season, suitable

breeding sites for amphibians, such as swamps, flooded

forests, forest ponds, and temporary puddles, formed

after rainfall around the rural complexes, and appeared

February 2022 | Volume 16 | Number 1 | e301

Amphibians of Kokolopori, central DR Congo

everywhere. The species compositions found in the wet

and dry seasons were similar, but Leptopelis christyi was

surprisingly found mainly in the dry season. This can be

attributed to its ecological preference and adaptation to

drier habitats (Amiet 2012). Some frogs were particularly

abundant in primary forests with flooded terrain or

swamps (e.g., Aubria masako, Hylambates verrucosus,

Phrynobatrachus cf. giorgii, and pipids), while others

were more commonly found in forests with high

canopy cover and numerous streams (e.g., Arthroleptis,

Cardioglossa, Leptopelis, Sclerophrys, and Ptychadena

aequiplicata). Hyperoliids and Amnirana lepus were

found mainly along streams at forest edges. Relatively

few species were encountered within rural complexes

(typically Ptychadena sp. aff. mascareniensis, and to a

lesser extent P. christyi and P. perreti, but the latter were

more commonly found in forests). Other species could

be found in peripheral farmbush (e.g., Afrixalus cf.

quadrivittatus and Amnirana cf. albolabris). Somewhat

surprisingly, no typical synanthropic bufonids were

found, although they are present almost everywhere in

the Congo (e.g., Sclerophrys gutturalis, S. pusilla, and

S. regularis). This is probably attributable to the well-

preserved forests of Kokolopori and only small roads/

paths leading into the area. In line with this finding,

the three Sclerophrys species recorded were obviously

restricted to forest habitats. Two of them could not be

identified to described species. In addition to the two

Sclerophrys toads, some other anurans, especially some

squeakers (Arthroleptis), also could not be identified to

species (Table 2). Some other taxa deserve taxonomic

revision and elevations from subspecies to species status

may be expected (e.g., Leptopelis and Hyperolius; Table

2). Our records of some species from Kokolopori represent

geographic range extensions (see Species Accounts).

Most importantly, Arthroleptis phrynoides, Leptopelis

calcaratus meridionalis, and L. ocellatus schiotzi are

reported here for the first time since their descriptions

(Laurent 1973, 1976a). Other interesting observations

worth mentioning include the biofluorescence observed

in male Hyperolius phantasticus boulengeri with Phase

J, which is consistent with the previous observations

on the presence of fluorescence in frogs with greenish-

translucent skin (Taboada et al. 2017a,b). To our

knowledge, this is the first documented case in an African

anuran.

Comparing our survey results with those of other

published surveys of the Central Congolian Forests

ecoregion (Penner and Rodel 2007: Lokutu; Schietz

2006: Kokolopori, Hyperolius only), our survey recorded

almost twice as many amphibian species as in Lokutu

and two more species of Hyperolius (although we

did not find one other species, see below). Penner and

Rodel (2007) recorded 21 species during 13 days and 14

nights in the wet season (October/November) in Lokutu,

which is only about 120 km north of Kokolopori. In

our two survey sessions in the wet season, we recorded

Amphib. Reptile Conserv.

30 and 31 species respectively over 15 days (Table 1).

The lower number of species observed in Lokutu may

be attributable to various factors, but most probably to

the more degraded habitats there. After re-evaluation

of the taxonomy, the species diversities recorded in

the Lokutu region and Kokolopori were similar. They

differed mainly in that several predominantly forest

genera (Arthroleptis, Aubria, Chiromantis, Congolius,

Hylambates, Hymenochirus, and Xenopus) were

not recorded in Lokutu, but on the other hand some

taxa known to occur commonly in disturbed areas

were recorded (e.g., synanthropic Sclerophrys toads,

Amnirana galamensis, Hoplobatrachus occipitalis, and

Ptychadena cf. taenioscelis). Penner and Rodel (2007)

also found a surprisingly low number of Hyperolius

species, with “H. cf. /ateralis” (occurring in eastern

DRC and East Africa; Channing and Rodel 2019) being

a likely misidentification. Hypero/ius in Kokolopori was

studied by Schigtz (2006). He did not record H. cf. langi

(although he probably discussed this taxon as H. kuligae,

from elsewhere in the central Congo), which was found

to be rare in our survey, and he misidentified H. cf.

cinnamomeoventris as light morphs of H. platyceps (for

more details, see Species Accounts). On the contrary,

he found one additional species, a single female of

H. brachiofasciatus Ahl, 1931 collected in farmbush in

syntopy with two other Hyperolius species. Since Schietz

found only the single individual and we did not record

this species, it seems likely that H. brachiofasciatus 1s

rare in Kokolopori. This taxon is very poorly known, with

few reports apart from its description (Ahl 1931; Jackson

and Blackburn 2007; Kielgast and Lotters 2011; Masudi

et al. 2019; Schiotz 2006). However, its morphology is

strikingly reminiscent of the H. tuberculatus complex, to

which it likely belongs, and may even represent an older

synonym of later described taxa such as H. ghesquieri

Laurent, 1943 or H. hutsebauti Laurent, 1956. This

hypothesis will require further verification.

Future descriptions and taxonomic revisions of the

above listed and discussed candidate species (potentially

new species to science; Table 2) will make an important

contribution to our understanding of the amphibian

fauna of Central Africa, and will further highlight the

biodiversity and biogeographic importance of the central

Congo. Despite the relatively low species richness, our

results suggest that the Central Congolian Lowland

Forests ecoregion harbors a unique and partially endemic

amphibian fauna that is distinct, to some extent, from the

anuran fauna to the north, east, and west of the wide arc

of the Congo River.

The Central Congolian forests are relatively well

preserved (Laporte et al. 2007; Mayaux etal. 2013), which

is a consequence of the region’s remoteness, limited

accessibility, and low human population. Kokolopori is

not directly connected by roads or rivers to large timber

markets such as those in Kisangani, Mbandaka, or

Kinshasa (pers. obs.), which supports the good condition

February 2022 | Volume 16 | Number 1 | e301

Badjedjea et al.

of the Kokolopori forests. However, some disturbances

from small-scale human activities are present in the

reserve, especially subsistence agriculture, including

slash-and-burn farming, and the presence of human

camps associated with poaching. To strike a balance

between the needs of the human population and nature

conservation, a new reserve delineation is currently

being prepared which focuses mainly on the primary

forest (A.L. Lokasola 2020, pers. comm.). Our results

can serve as a basis for future monitoring of amphibians,

assessment of their population status, and conservation

management planning and strategies in the Kokolopori

Bonobo Nature Reserve and surrounding regions. The

community-based conservation in Kokolopori may

provide a good example for other areas in the Central

Congolian forests and throughout the Democratic

Republic of the Congo.

Acknowledgments.—We thank the Kokolopori

community and especially A.L. Lokasola (Kinshasa/

Yetee) for inviting us to conduct an amphibian survey

of the Kokolopori Bonobo Nature Reserve. We would

also like to thank the many curators and technicians at

several museums for access to materials and resources,

in particular D. Meirte and G. Cael (RMCA), A. Ohler

(MNHN), D. Kizirian and L. Vonnahme (AMNH), J.

Rosado (MCZ), M.-O. Rodel, and F. Tillack (ZMB),

and B. Clarke and J.W. Streicher (BMNH). We also

thank A. Hanova, F. Snitily, and T. Neéas for their help

in the laboratory and fruitful discussions. We are further

grateful to Alan Channing and Eli Greenbaum for their

constructive comments. This research was supported by

the International Foundation for Science (Stockholm),

through a grant to G.B. (No. I-1-D-6074-1). V.G. was

supported by the Ministry of Culture of the Czech

Republic (DKRVO = 2019-2023/6.VII.d, National

Museum, 00023272) and institutional support from the

IVB CAS (RVO: 68081766).

Literature Cited

Ahl E. 1931. Amphibia, Anura III, Polypedatidae. Das

Tierreich 55: 1—477.

Almquist LA, LokasolaAL, Coxe SJ, Hurley MJ, Scherlis

JS. 2010. Kokolopori and the Bonobo Peace Forest in

the Democratic Republic of the Congo: prioritizing

the local in conservation practice. Pp. 311-326 In:

Indigenous Peoples and Conservation: From Rights

to Resource Management. Editors, Walker Painemilla

K, Rylands AB, Woofter A, Hughes C. Conservation

International, Arlington, Virginia, USA. 326 p.

Amiet J-L. 1979 “1978.” A propos d’ Hyperolius platyceps

(Boulenger), H. kuligae Mertens et H. adametzi Ahl

(Amphibiens Anoures). Annales de la Faculte des

Sciences du Cameroun 25: 221-256.

Amiet J-L. 2012. Les Rainettes du Cameroun (Amphibiens

Amphib. Reptile Conserv.

Anoures). J.-L. Amiet and La Nef des Livres, Nyons

and Saint-Nazaire, France. 591 p.

Amiet J-L, Goutte S. 2017. Chants d’Amphibiens du

Cameroun. Imprimerie Chirat, Saint-Just-la-Pendue,

France. 280 p.

Anthony NM, Atteke C, Bruford MW, Dallmeier F,

Freedman A, Hardy O, Ibrahim B, Jeffery KJ, Johnson

M, Lahm SA, et al. 2015. Evolution and conservation

of Central African biodiversity: priorities for future

research and education in the Congo Basin and Gulf

of Guinea. Biotropica 47: 6-17.

Badjedjea GB, Jocqué R, Masudi FM, Rédel M-O,

Burger M, Gvozdik V, Pauwels OSG. 2019. Frog-

eating spiders in the Afrotropics: an analysis of

published and new cases. Bulletin of the Chicago

Herpetological Society 54: 57-63.

Basualdo CV. 2011. Choosing the best non-parametric

richness estimator for benthic macroinvertebrates

databases. Revista de la Sociedad Entomoldogica

Argentina 70: 27-38.

Bell RC, Drewes RC, Channing A, Gvozdik V, Kielgast

J, Lotters S, Stuart BL, Zamudio KR. 2015. Overseas

dispersal of Hyperolius Reed Frogs from Central

Africa to the oceanic islands of Sao Tomé and

Principe. Journal of Biogeography 42: 65-75.

Bell RC, Parra JL, Badjedjea G, Bare; MF, Blackburn

DC, Burger M, Channing A, Dehling JM, Greenbaum

E, Gvozdik V, et al. 2017. Idiosyncratic responses

to climate-driven forest fragmentation and marine

incursions in Reed Frogs from Central Africa and

the Gulf of Guinea Islands. Molecular Ecology 26:

5,223-5,244.

Bittencourt-Silva GB. 2019. A herpetological survey of

western Zambia. Amphibian & Reptile Conservation

13(2) [Special Section]: 1-28 (e181).

Blackburn DC. 2008. Biogeography and evolution of

body size and life history of African frogs: phylogeny

of Squeakers (Arthroleptis) and Long-fingered Frogs

(Cardioglossa) estimated from mitochondrial data.

Molecular Phylogenetics and Evolution 49: 806-826.

Blackburn DC, Nielsen SV, Ghose SL, Burger M,

Gonwouo LN, Greenbaum E, Gvozdik V, Hirschfeld

M, Kouete MT, Kusamba C, et al. 2021. Phylogeny

of African Long-fingered Frogs (Arthroleptidae:

Cardioglossa) reveals recent allopatric divergences in

coloration. /chthyology & Herpetology 109: 728-742.

Boulenger GA. 1899. Descriptions of new batrachians

in the collection of the British Museum (Natural

History). The Annals and Magazine of Natural History

7" Series 3: 273-277.

Boulenger GA. 1919. Batraciens et reptiles recueillis par

le Dr. C. Christy au Congo Belge dans les Districts de

Stanleyville, Haut-Uelé et Ituri en 1912-1914. Revue

Zoologique Africaine 7: 1-29.

Burgess N, Hales JDA, Underwood E, Dinerstein E, Olson

D, Itoua I, Schipper J, Ricketts T, Newman K. 2004.

Terrestrial Ecoregions of Africa and Madagascar. A

February 2022 | Volume 16 | Number 1 | e301

Amphibians of Kokolopori, central DR Congo

Conservation Assessment. Island Press, Washington,

DC, USA. 497 p.

Burnham KP, Overton WS. 1978. Estimation of the size

of a closed population when capture probabilities vary

among animals. Biometrika 65: 625-633.

Burnham KP, Overton WS. 1979. Robust estimation

of population size when capture probabilities vary

among animals. Ecology 60: 927-936.

Channing A, Rodel M-O. 2019. Field Guide to the Frogs

and Other Amphibians of Africa. Struik Nature, Cape

Town, South Africa. 408 p.

Channing A, Rodel M-O, Channing J. 2012. Tadpoles of

Africa: the Biology and Identification of All Known

Tadpoles in sub-Saharan Africa. Edition Chimaira,

Frankfurt am Main, Germany. 402 p.

Chao A. 1987. Estimating the population size for capture-

recapture data with unequal catchability. Biometrics

43: 783-791.

Chao A, Lee SM, Jeng SL. 1992. Estimating population

size for capture-recapture data when capture

probabilities vary by time and individual animal.

Biometrics 48: 201-216.

Chazdon RL, Colwell RK, Denslow JS, Guariguata

MR. 1998. Statistical methods for estimating species

richness of woody regeneration in primary and

secondary rain forests of NE Costa Rica. Pp. 285—309

In: Forest Biodiversity Research, Monitoring and

Modeling: Conceptual Background and Old-World

Case Studies. Editors, Dallmeier F, Comiskey JA.

Parthenon Publishing, Paris, France. 671 p.

Chifundera Kusamba Z. 2019. Using diversity indices

for identifying the priority sites for herpetofauna

conservation in the Democratic Republic of the

Congo. Nature Conservation Research 4: 13-33.

Chifundera Kusamba Z, Nagy Z, Greenbaum E, Kielgast

J, Gvozdik V, Mebert K. 2014. L’herpetofaune de

la Cuvette Centrale en RD Congo. Pp. 96-97 In: 1”

International Conference on Biodiversity in the Congo

Basin. 6-10 June 2014, Kisangani, Democratic

Republic of the Congo. Consortium Congo 2010 and

Centre de Surveillance de la Biodiversité, Kinshasa,

Democratic Republic of the Congo. 222 p.

Colwell RK. 2013. EstimateS: Statistical Estimation of

Species Richness and Shared Species from Samples.

Version 9. User’s Guide and application. Available:

http://purl.oclc.org/estimates [Accessed: 14 July

2021].

Colwell RK, Elsensohn JE. 2014. EstimateS turns

20: statistical estimation of species richness and

shared species from samples, with non-parametric

extrapolation. Ecography 37: 609-613.

Colyn M, Gautier-Hion A, Verheyen W. 1991. A re-

appraisal of palaeoenvironmental history in Central

Africa: evidence for a major fluvial refuge in the Zaire

Basin. Journal of Biogeography 18: 403-407.

de Queiroz K. 2020. An updated concept of subspecies

resolves a dispute about the taxonomy of incompletely

Amphib. Reptile Conserv.

separated lineages. Herpetological Review 51: 459-

461.

De Witte GF. 1930. Liste des batraciens du Congo Belge

(collection du Musée du Congo Belge a Tervuren).

Premiere partie. Revue de Zoologie et de Botanique

Africaines 19: 232-274.

Deichmann JL, Mulcahy DG, Vanthomme H, Tobi E,

Wynn AH, Zimkus BM, McDiarmid RW. 2017.

How many species and under what names? Using

DNA barcoding and GenBank data for west Central

African amphibian conservation. PLoS ONE 12(11).:

00187283.

Dewynter M, Frétey T. 2019. Liste taxonomique

commentée et catalogue illustré des Amphibiens du

Gabon. Les Cahiers de la Fondation Biotope 27:

1-84.

Dinerstein E, Olson D, Joshi A, Vynne C, Burgess, ND,

Wikramanayake E, Hahn N, Palminter1 S, Hedao P,

Noss R, et al. 2017. An ecoregion-based approach to

protecting half the terrestrial realm. BioScience 67:

534-545.

Dubois A, Ohler A, Pyron RA. 2021. New concepts and

methods for phylogenetic taxonomy and nomenclature

in zoology, exemplified by a new ranked cladonomy

of recent amphibians (Lissamphibia). Megataxa 5:

1-738.

Ernst R, Lautenschlager T, Branquima MF, Holting M.

2020. At the edge of extinction: a first herpetological

assessment of the proposed Serra do Pingano

Rainforest National Park in Uige Province, northern

Angola. Zoosystematics and Evolution 96: 237-262.

Evans BJ, Carter TF, Greenbaum E, Gvozdik V, Kelley

DB, McLaughlin PJ, Pauwels OSG, Portik DM,

Stanley EL, Tinsley RC, et al. 2015. Genetics,

morphology, advertisement calls, and_ historical

records distinguish six new polyploid species of

African Clawed Frog (Xenopus, Pipidae) from West

and Central Africa. PLoS ONE 10(12): e0142823.

Evans BJ, Gansauge M-T, Stanley EL, Furman BLS,

Cauret CMS, Ofori-Boateng C, Gvozdik V, Streicher

JW, Greenbaum E, Tinsley RC, et al. 2019. Xenopus

fraseri: Mr. Fraser, where did your frog come from?

PLoS ONE 14(9): e0220892.

FACET. 2010. Foréts d’Afrique Centrale Evaluées par

Télédétection. Etendue et Perte du Couvert Forestier

en République Démocratique du Congo de 2000 a

2010. South Dakota State University, Brookings,

South Dakota, USA. Available: https://osfac.net/

data-products/facet/facet-dr-congo [Accessed: 18

December 2020].

Fligel TJ, Eckardt FD, Cotterill FPD. 2015. The present-

day drainage patterns of the Congo River system and

their Neogene evolution. Pp. 315-337 In: Geology

and Resource Potential of the Congo Basin. Editors,

de Wit MJ, Guillocheau F, de Wit MCJ. Springer-

Verlag, Berlin, Germany. 417 p.

Frétey T, Dewynter M, Blanc CP. 2011. Amphibiens

February 2022 | Volume 16 | Number 1 | e301

Badjedjea et al.

d’Afrique Centrale et d’Angola. Clé de Détermination

Illustrée des Amphibiens du Gabon et du Mbini.

Biotope, Meéze (Collection Parthénope) and Muséum

National d’ Histoire Naturelle, Paris, France. 232 p.

Frost DR. 2021. Amphibian Species of the World: an

Online Reference. Version 6.1. American Museum

of Natural History, New York, New York, USA.

Available: — https://amphibiansoftheworld.amnh.org/

index.php [Accessed: 18 April 2021].

Gosner KL. 1960. A simplified table for staging anuran

embryos and larvae with notes on identification.

Herpetologica 16: 183-190.

Greenbaum E. 2017 “2018.” Emerald Labyrinth: a

Scientist's Adventures in the Jungles of the Congo.

ForeEdge (imprint of University Press of New

England), Lebanon, New Hampshire, USA. 329 p.

Greenbaum E, Conkey N, Kusamba Chifundera Z,

Pramuk J, Carr J, Rodel M, Jackson K, Kielgast

J, Nagy Z, Penner J. 2014. Systematics of Congo

Basin true toads (Anura: Bufonidae: Amietophrynus)

reveals widespread cryptic speciation. Pp. 72—73 In:

1” International Conference on Biodiversity in the

Congo Basin. 6—10 June 2014, Kisangani, Democratic

Republic of the Congo. Consortium Congo 2010 and

Centre de Surveillance de la Biodiversité, Kinshasa,

Democratic Republic of the Congo. 222 p.

Gvozdik V, Chifundera Kusamba Z. 2014. Amphibians

and reptiles recorded in the region of the Eastern

Congolian swamp forests to the east of Lake Mai-

Ndombe, Democratic Republic of the Congo.

Pp. 160-161 In: /* International Conference on

Biodiversity in the Congo Basin. 6-10 June 2014,

Kisangani, Democratic Republic of the Congo.

Consortium Congo 2010 and Centre de Surveillance

de la Biodiversité, Kinshasa, Democratic Republic of

the Congo. 222 p.

Guibé J, Lamotte M. 1958. Les Ptychadena (Batraciens,

Ranidés) du Cameroun. Bulletin de | ’Institut Francaise

d’Afrique Noire, Série A, Sciences Naturelles 20:

1,448-1,463.

Hart JA, Detwiler KM, Gilbert CC, Burrell AS, Fuller

JL, Emetshu M, Hart TB, Vosper A, Sargis EJ, Tosi

AJ. 2012. Lesula: A new species of Cercopithecus

monkey endemic to the Democratic Republic of

Congo and implications for conservation of Congo’s

Central Basin. PLoS ONE 7(9): e44271.

Herrmann H-W, Bohme W, Herrmann PA, Plath M,

Schmitz A, Solbach M. 2005. African biodiversity

hotspots: the amphibians of Mt. Nlonako, Cameroon.

Salamandra 41: 61-81.

Hillis DM. 2019. Species delimitation in herpetology.

Journal of Herpetology 53: 3-12.

Hillis DM. 2020. The detection and naming of geographic

variation within species. Herpetological Review 51:

52-56.

Hirschfeld M, Blackburn DC, Burger M, Greenbaum

E, Zassi-Boulou A-G, Rodel M-O. 2015. Two

Amphib. Reptile Conserv.

new species of Long-fingered Frogs of the genus

Cardioglossa (Anura: Arthroleptidae) from Central

African rainforests. African Journal of Herpetology

64: 81-102.

Hulselmans JLJ. 1977. Further notes on African

Bufonidae, with descriptions of new species and

subspecies (Amphibia, Bufonidae). Revue de Zoologie

Africaines 91: 512-524.

IUCN SSC Amphibian Specialist Group. 2017.

Arthroleptis tuberosus. The TUCN Red List of

Threatened Species 2017: e.T54390A 18365432.

IUCN SSC Amphibian Specialist Group. 2019.

Ptychadena aequiplicata. The IUCN Red List of

Threatened Species 2019: e.T58489A 18398710.

IUCN SSC Amphibian Specialist Group. 2020.

Ptychadena perreti (amended version of 2017

assessment). The IUCN Red List of Threatened

Species 2020: e.T58518A177065443.

Jackson K, Blackburn DC. 2007. The amphibians and

reptiles of Nouabale-Ndoki National Park, Republic

of Congo (Brazzaville). Salamandra 43: 149-164.

Joger U. 1982. Zur Herpetofaunistik Kameruns (II).

Bonner Zoologische Beitrdge 33: 313-342.

Jongsma GFM, Bare) MF, Barratt CD, Burger M,

Conradie W, Ernst R, Greenbaum E, Hirschfeld M,

Leaché AD, Penner J, et al. 2018. Diversity and

biogeography of frogs in the genus Amnirana (Anura:

Ranidae) across sub-Saharan Africa. Molecular

Phylogenetics and Evolution 120: 274-285.

Jongsma GFM, Tobi E, Dixon-MacCallum GP, Bamba-

Kaya A, Yoga J-A, Mbega J-D, Mve Beh J-H, Emrich

AM, Blackburn DC. 2017. Amphibians of Haut-

Ogooué Province, southeastern Gabon. Amphibian &

Reptile Conservation 11(1) [Special Section]: 1—23

(e144).

Kielgast J, Lotters S. 2011. The green heart of Africa is a

blind spot in herpetology. FrogLog 97: 16-17.

Kielgast J, Nagy Z, Chifundera Kusamba Z, Lotana

Lokasola A, Bodongola A, Mebert K, Loetters

S. 2014. Knowledge-gaps and novelty in_ the

amphibian diversity of the Democratic Republic of

the Congo. Pp. 190 In: /* International Conference

on Biodiversity in the Congo Basin. 6-10 June 2014,

Kisangani, Democratic Republic of the Congo.

Consortium Congo 2010 and Centre de Surveillance

de la Biodiversité, Kinshasa, Democratic Republic of

the Congo. 222 p.

Kohler J, Scheelke K, Schick S, Veith M, Lotters S. 2005.

Contribution to the taxonomy of hyperoliid frogs

(Amphibia: Anura: Hyperoliidae): advertisement

calls of twelve species from East and Central Africa.

African Zoology 40: 127-142.

Kunz K. 2007. Der Zwergkrallenfrosch: Hymenochirus

boettgeri. 2" Edition. Natur und Tier Verlag, Munster,

Germany. 64 p.

Laporte NT, Stabach JA, Grosch R, Lin TS, Goetz SJ.

2007. Expansion of industrial logging in Central

February 2022 | Volume 16 | Number 1 | e301

Amphibians of Kokolopori, central DR Congo

Africa. Science 316: 1,451.

Largen MJ, Dowsett-Lemaire F. 1991. Amphibians

(Anura) from the Kouilou River basin, République du

Congo. Zauraco Research Report 4: 145-168.

Laurent R. 1941. Contribution a la systématique du genre

Hyperolius Rapp (Batraciens). Revue de Zoologie et

de Botanique Africaines 34: 149-167.

Laurent R. 1943. Les Hyperolius (Batraciens) du Musée

du Congo. Annales du Musée du Congo Belge. C.

Zoologie, Série I 4: 61-140.

Laurent R. 1950. Genres Afrixalus et Hyperolius

(Amphibia Salientia). Exploration du Parc National

Albert. Mission G. F: de Witte (1933-1935) 64: 5-120.

Laurent R. 1952. Reptiles et Batraciens nouveaux de la

région des Grands Lacs africains. Revue de Zoologie

et de Botanique Africaines 46: 269-279.

Laurent RF. 1957. Genres Afrixalus et Hyperolius

(Amphibia Salientia). Exploration du Parc National

de I’Upemba. Mission G. F: de Witte, en collaboration

avec W. Adam, A. Janssens, L. van Meel et R. Verheyen

(1946-1949) 42: 3-47.

Laurent RF. 1972. Amphibiens. Exploration du Parc

National des Virunga. Deuxiéme Série 22: 1-125.

Laurent RF. 1973. Le genre Leptopelis Gunther

(Salientia) au Zaire. Annales du Musée Royal de

l’Afrique Centrale. Tervuren, Belgique. Série in-8°

Sciences Zoologique 202: 1-62.

Laurent RF. 1976a. Deux Amphibiens nouveaux du

Zaire. Revue de Zoologie Africaine 90: 528-546.

Laurent RF 1976b. Les” genres Cryptothylax,

Phlyctimantis, et Kassina au Zaire. Annales du Musée

Royal de I’ Afrique Centrale. Tervuren, Belgique. Série

in-8°, Sciences Zoologique 213: 1-67.

Laurent RF. 1979. Description de deux Hyperolius

nouveaux du Sankuru = (Zaire) (Amphibia,

Hyperoliidae). Revue de Zoologie Africaines 93: 779-

TOM,

Laurent R. 1982. Le genre Afrixalus Laurent

(Hyperoliidae) en Afrique centrale. Annales du Musée

Royal de I’ Afrique Centrale. Tervuren, Belgique. Série

in-8°, Sciences Zoologique 235: 1-58.

Leaché AD, Portik DM, Rivera D, R6del M-O, Penner

J, Gvozdik V, Greenbaum E, Jongsma GFM, Ofori-

Boateng C, Burger M, et al. 2019. Exploring rain forest

diversification using demographic model testing in the

African Foam-nest Treefrog, Chiromantis rufescens.

Journal of Biogeography 46: 2,706—2,721.

Liedtke HC, Muller H, Hafner J, Penner J, Gower DJ,

Mazuch T, Rodel M-O, Loader SP. 2017. Terrestrial

reproduction as an adaptation to steep terrain in

African toads. Proceedings of the Royal Society B

284: 20162598.

Liedtke HC, Miller H, Rodel M-O, Menegon M, Gonwouo

LN, Bare} MF, Gvozdik V, Schmitz A, Channing A,

Nagel P, et al. 2016. No ecological opportunity signal

ona continental scale? Diversification and life-history

evolution of African true toads (Anura: Bufonidae).

Amphib. Reptile Conserv.

Evolution 70: 1,717-1,733.

Lokasola AL, Botshuna CL, Badjejea GB, Gembu CT.

2017. Distributional data of the lizard fauna (Sauria) of

the Maringa-Lopori-Wamba Landscape, Democratic

Republic of the Congo. Journal of Environmental

Science and Engineering B 6: 151-159.

Maley J. 1987. Fragmentation de la forét dense humide

africaine et extension des biotopes montagnards au

Quaternaire recent. Nouvelles données polliniques

et chronologiques. Implications paléoclimatiques et

biogéographiques. Pp. 307-334 In: Palaeoecology of

Africa and the Surrounding Islands, Volume 18. Editor,

Coetzee JA. Balkema, Rotterdam, Netherlands. 486 p.

Maley J. 1991. The African rain forest vegetation and

palaeoenvironments during the late Quaternary.

Climatic Change 19: 79-98.

Maley J. 1996. TheA frican rain forest: main characteristics

of changes in vegetation and climate from the Upper

Cretaceous to the Quaternary. Proceedings of the

Royal Society of Edinburgh Section B 104: 31-73.

Marques MP, Ceriaco LMP, Blackburn DC, Bauer AM.

2018. Diversity and distribution of the amphibians

and terrestrial reptiles of Angola. Atlas of historical

and bibliographic records (1840-2017). Proceedings

of the California Academy of Sciences, Series 4

65(Supplement IT): 1-501.

Masudi FMM, Banda PKG, Kusamba ZC, Badjedjea

GB, Sebe JRK, Lokasola AL, Ewango C, Gembu

GCT, Dudu A. 2019. Preliminary data on amphibian

diversity of the Okapi Wildlife Reserve (RFO) in

Democratic Republic of the Congo. American Journal

of Zoology 2: 38-43.

Mayaux P, Pekel J-F, Desclée B, Donnay F, Lupi A,

Achard F, Clerici M, Bodart C, Brink A, Nasi R, et al.

2013. State and evolution of the African rainforests

between 1990 and 2010. Philosophical Transactions

of the Royal Society B 368: 20120300.

Mulotwa M, Louette M, Dudu A, Upoki A, Fuller

RA. 2010. Congo Peafowl use both primary and

regenerating forest in Salonga National Park,

Democratic Republic of Congo. Ostrich 81: 1-6.

Neé€as T, Badjedjea G, Vopalensky M, GvoZzdik V. 2021.

Congolius, anew genus of African Reed Frog endemic

to the central Congo: a potential case of convergent

evolution. Scientific Reports 11: 8,338.

Nicolas V, Barriere P, Colyn M. 2003. Impact of

removal pitfall trapping on the community of shrews

(Mammalia: Soricidae) in two African tropical forest

sites. Mammalia 67: 133-138.

Nneji LM, Adeola AC, Adedeji BE, Olatunde O, Onadeko

AB, Eniang EA, Ayoola AO, Adentyi AV, OkeyoyinA,

Oladipo SO, et al. 2021. Species richness, distribution

pattern, and conservation status of amphibians in

Cross River National Park, south-eastern Nigeria.

Biologia 76: 2,573-2,588.

Nneji LM, Adeola AC, OkeyoyinA, Onadeko AB, Eniang

EA, Adeyi AO, Oladipo SO, Adedeji BE, Usongo

February 2022 | Volume 16 | Number 1 | e301

Badjedjea et al.

JY, Andem AB, et al. 2019. First record of Foulassi

Screeching Frog, Arthroleptis adelphus (Perret,

1966) (Anura, Arthroleptidae, Arthroleptinae), from

Nigeria, with notes on its phylogenetic position.

Check List 15: 253-258.

Noble GK. 1924. Contributions of the herpetology of

the Belgian Congo based on the collection of the

American Museum Congo Expedition, 1909-1915.

Part II. Amphibia. Bulletin of the American Museum

of Natural History 49: 147-347.

Nussbaum RA, Pfrender ME. 1998. Revision of

the African caecilian genus Schistometopum

Parker (Amphibia: Gymnophiona: Caeciltidae).

Miscellaneous Publications, Museum of Zoology,

University of Michigan 187: 1-32.

O’Connor C. 2015. Herpetological Study for Feronia,

Lokutu Oil Palm Plantation. High Conservation Value

Assessment Report. Digby Wells Environmental,

Randburg, South Africa. 44 p. Available: https://

feronia.com/uploads/20 1 8-02-08/v3-herpetological-

study-lokutu-final90519.pdf [Accessed: 23 September

2020].

Ohler A, Dubois A. 2016. The identity of the South African

toad Sclerophrys capensis Tschudi, 1838 (Amphibia,

Anura). PeerJ 4: 1-13 (e1553).

Ohler A, Kazadi M. 1990 “1989.” Description d’une

nouvelle espéece du genre Aubria Boulenger, 1917

(Amphibiens, Anoures) et redescription du type

dV Aubria subsigillata (A. Dumeril, 1856). Alytes 8:

25-40.

Pauwels OSG, Rodel M-O. 2007. Amphibians and

national parks in Gabon, western Central Africa.

Herpetozoa 19: 135-148.

Penner J, Rodel M-O. 2007. Amphibians and reptiles of

Lokutu. Pp. 37-41 In: A Rapid Biological Assessmentof

Lokutu, Democratic Republic of Congo. RAP Bulletin

of Biological Assessment 46. Editors, Butynski TM,

McCullough J. Conservation International, Arlington,

Virginia, USA. 81 p.

Perret J-L. 1976a. Revision des amphibiens africains et

principalement des types, conservés au Musée Bocage

de Lisbonne. Arquivos do Museu Bocage. Série 2 6:

15-34.

Perret J-L. 1976b. Identité de quelques Afrixalus

(Amphibia, Salientia, Hyperoliidae). Bulletin de la

Société Neuchdteloise des Sciences Naturelles 99:

19-28.

Pickersgill M. 2007. A_ redefinition of Afrixalus

fulvovittatus (Cope, 1860) and Afrixalus vittiger

(Peters, 1876) (Amphibia, Anura, Hyperoliidae).

African Journal of Herpetology 56: 23-37.

Plana V. 2004. Mechanisms and tempo of evolution in the

African Guineo-Congolian rainforest. Philosophical

Transactions of the Royal Society of London B 359:

1,585—1,594.

Portik DM, Bell RC, Blackburn DC, Bauer AM, Barratt

CD, Branch WR, Burger M, Channing A, Colston TJ,

Amphib. Reptile Conserv.

Conradie W, et al. 2019. Sexual dichromatism drives

diversification within a major radiation of African

amphibians. Systematic Biology 68: 859-875.

Portik DM, Jongsma GFM, Kouete MT, Scheinberg LA,

Freiermuth B, Tapondjou WP, Blackburn DC. 2016.

A survey of amphibians and reptiles in the foothills

of Mount Kupe, Cameroon. Amphibian & Reptile

Conservation 10(2) [Special Section]: 37-67 (e131).

Rabb GB, Rabb MS. 1963. On the behavior and breeding

biology of the African pipid frog Hymenochirus

boettgeri. Zeitschrift fiir Tierpsychologie 20: 215-241.

Roédel M-O. 2000. Herpetofauna of West Africa. Volume

I; Amphibians of the West African Savanna. Edition

Chimaira, Frankfurt am Main, Germany. 332 p.

Sanchez- Vialas A, Calvo-Revuelta M, Castroviejo-Fisher

S, De la Riva I. 2020. Synopsis of the amphibians of

Equatorial Guinea based upon the authors’ field work

and Spanish natural history collections. Proceedings

of the California Academy of Sciences, Series 4 66:

137-230.

Schick S, Kielgast J, Rodder D, Muchai V, Burger M,

Lotters S. 2010. New species of Reed Frog from

the Congo basin with a discussion of paraphyly in

Cinnamon-belly Reed Frogs. Zootaxa 2501: 23-36.

Schigtz A. 1967. The treefrogs (Rhacophoridae) of West

Africa. Spolia Zoologica Musei Hauniensis 25: 1-346.

Schiotz A. 1999. Treefrogs of Africa. Edition Chimaira,

Frankfurt am Main, Germany. 351 p.

Schiotz A. 2006. Notes on the genus Hyperolius (Anura,

Hyperoliidae) in central République Démocratique du

Congo. Alytes 24: 40-60.

Smith EP, van Belle G. 1984. Nonparametric estimation

of species richness. Biometrics 40: 119-129.

Stanton DWG, Hart J, Vosper A, Kumpel NF, Wang JL,

Ewen JG, Bruford MW. 2016. Non-invasive genetic

identification confirms the presence of the Endangered

Okapi, Okapia johnstoni, south-west of the Congo

River. Oryx 50: 134-137.

Surbeck M, Coxe S, Lokasola AL. 2017. Lonoa: the

establishment of a permanent field site for behavioral

research on Bonobos in the Kokolopori Bonobo

Reserve. Pan Africa News 24: 13-15.

Taboada C, Brunetti AE, Pedron FN, Carnevale Neto F,

Estrin DA, Bari SE, Chemes LB, Peporine Lopes N,

Lagorio MG, Faivovich J. 2017a. Naturally occurring

fluorescence in frogs. Proceedings of the National

Academy of Sciences of the United States of America

114: 3,672-3,677.

Taboada C, Brunetti AE, Alexandre C, Lagorio MG,

FaivovichJ.2017b. Fluorescent frogs: a herpetological

perspective. South American Journal of Hepretology

12: 1-13.

Tandy M, Keith R. 1972. Bufo of Africa. Pp. 119-170

In: Evolution in the Genus Bufo. Editor, Blair WF.

University of Texas Press, Austin, Texas, USA. 459 p.

Tolley KA, Alexander GJ, Branch WR, Bowles P, Maritz

B. 2016. Conservation status and threats for African

February 2022 | Volume 16 | Number 1 | e301

Amphibians of Kokolopori, central DR Congo

reptiles. Biological Conservation 204: 63-71.

Vancutsem C, Pekel J-F, Evrard C, Malaisse F, Defourny

P. 2009. Mapping and characterizing the vegetation

types of the Democratic Republic of Congo using

SPOT VEGETATION time series. /nternational

Journal of Applied Earth Observation and

Geoinformation 11: 62-76.

Van de Perre F, Leirs H, Verheyen E. 2019. Paleoclimate,

ecoregion size, and degree of isolation explain

regional biodiversity differences among terrestrial

vertebrates within the Congo Basin. Belgian Journal

of Zoology 149: 23-42.

Van Krunkelsven E, Bila-Isia I, Draulans D. 2000. A

survey of Bonobos and other large mammals in the

Salonga National Park, Democratic Republic of

Congo. Oryx 34: 180-187.

Vences M, Kosuch J, Rodel M-O, Lotters S, Channing

A, Glaw F, Bohme W. 2004. Phylogeography of

Ptychadena mascareniensis suggests transoceanic

dispersal in a widespread African-Malagasy frog

lineage. Journal of Biogeography 31: 593-601.

Congolian rainforests.

Amphib. Reptile Conserv.

Vigny C. 1979. Morphologie larvaire de 12 espeéces et

sous-especes du genre Xenopus. Revue Suisse de

Zoologie 86: 877-891.

Williams VL, Witkowski ETF, Balkwill K. 2007. The

use of incidence-based species richness estimators,

species accumulation curves, and similarity measures

to appraise ethnobotanical inventories from South

Africa. Biodiversity and Conservation 16: 2,495—

25 V3"

Zimkus BM, Larson JG. 2013. Assessment of the

amphibians of Batéké Plateau National Park,

Gabon, including results of chytrid pathogen tests.

Salamandra 49: 159-170.

Zimkus BM, Lawson LP, Bare} MF, Barratt CD,

Channing A, Dash KM, Dehling JM, Du Preez L,

Gehring P-S, Greenbaum E, et al. 2017. Leapfrogging

into new territory: how Mascarene Ridged Frogs

diversified across Africa and Madagascar to maintain

their ecological niche. Molecular Phylogenetics and

Evolution 106: 254—269.

Gabriel Badjedjea is a Ph.D. student in the Department of Aquatic Ecology, Faculty of Sciences,

University of Kisangani (UNIKIS), Democratic Republic of the Congo. He is currently responsible for the

herpetological collection at the Biodiversity Monitoring Center, UNIKIS. His Ph.D. research focuses on

amphibian assemblages and diversity in the central and northeastern Congo Basin.

Franck M. Masudi recently completed his M.Sc. degree in Zoological Sciences in the Faculty of Sciences,

University of Kisangani, Democratic Republic of the Congo. He works in herpetology, with interests in

diversity, ecology, and taxonomy. His Ph.D. project will focus on the ecology and taxonomy of amphibians

in protected areas of the Democratic Republic of the Congo.

Benjamin Dudu Akaibe is Professor Emeritus at the Faculty of Sciences, University of Kisangani,

Democratic Republic of the Congo. His research focus includes the diversity and ecology of African

rainforest mammals; rodents of the central Congo Basin; applied ecology for sustainable forest resource

management, including domestic and wild small mammal husbandry; conservation biology of the Congolian

forest ecosystem; and bushmeat and zoonoses. He is the Director of the Biodiversity Monitoring Center

(Centre de Surveillance de la Biodiversité), UNIKIS, and leads a team of experts working in applied ecology,

pest control, bushmeat trade monitoring, and the sustainable use of renewable resources in the Congo Basin.

Vaclav Gvozdik is a herpetologist working at the Institute of Vertebrate Biology of the Czech Academy

of Sciences (Brno) and the National Museum (Prague), Czech Republic. Vaclav is interested in the

phylogeography, diversity, and evolution of amphibians and reptiles of tropical Africa and the Western

Palearctic. He has particular experience with the herpetofauna of Central Africa and a special interest in the

February 2022 | Volume 16 | Number 1 | e301

Official journal website:

amphibian-reptile-conservation.org

Amphibian & Reptile Conservation

16(1) [General Section]: 71-75 (e302).

Sex hormones in the Axolotl, Ambystoma mexicanum:

potential method for sex determination

13Ilsabel A. Veith, and 2’Chester R. Figiel, Jr.

'School of Biological Sciences, Georgia Institute of Technology, Atlanta, Georgia, USA **Warm Springs Fish Technology Center, U.S. Fish and

Wildlife Service, Warm Springs, Georgia, USA

Abstract.—Axolotls, Ambystoma mexicanum (Shaw and Nodder 1798), are a Critically Endangered, aquatic

salamander species. Despite being imperiled in the wild, laboratory colonies are bred extensively for

developmental and embryological research. Verifying the sex of young-of-year would benefit researchers and

culturists especially when organisms take a long time to develop or become sexually mature. In this study,

we investigated whether steroid hormone metabolites (i.e., testosterone, 17B-estradiol) could be measured

in three age classes of axolotls (classified as juvenile, sub-adults, and adults). Our objectives were firstly to

validate whether significant levels of steroid metabolites could be detected in axolotis at various developmental

stages using a previously unexplored method of extraction. Secondly, if significant levels of hormones were

detected, could we differentiate between the sexes of adults by examining differences in the concentrations of

steroid hormone metabolites? Steroid hormone analysis of tissue samples determined both testosterone and

17B-estradiol were present in detectable concentrations in all age classes. There was no significant difference

in estradiol between females and males (t [11] = 0.89, p = 0.3881), however testosterone concentrations were

approaching significance (t [10] = 1.81, p = 0.0569) with females having over twice as much testosterone (x

= 0.36 + 0.07 ng/mL, n = 7) compared to males (x = 0.17 + 0.04 ng/mL, n = 5). There was not a significant

difference (t [10] = 1.81, p = 0.1112) in the ratio of testosterone to estradiol between sexes. The presence of

these hormones in earlier developmental stages was also confirmed providing the prospect that hormone

changes can be tracked over the course of sexual development.

Keywords. Estradiol, testosterone, salamanders, conservation, enzyme immunoassays, Mexico

Citation: Veith IA, Figiel CR Jr. 2022. Sex hormones in the Axolotl, Ambystoma mexicanum: potential method for sex determination. Amphibian &

Reptile Conservation 16(1) [General Section]: 71-75 (e302).

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.

Accepted: 30 October 2021; Published: 15 February 2022

Introduction for axolotls and other salamanders, knowing the sex

of individuals early in their development can play an

Ambystoma mexicanum (Shaw and Nodder 1798), the — essential role in conservation planning. Determining

axolotl, is native to Lake Xochimilco, a high-altitude | sex at a younger age benefits researchers and culturists,

lake in the Central Valley of Mexico. The expansion — especially when individuals take a long time to develop

of agriculture and human development, compounded and become sexually mature. Additionally, verifying the

with the effects of deteriorating water quality and sex of young-of-year can be crucial in understanding

the introduction of non-native fish, has decimated A. demographics in field studies. Having to wait until

mexicanum populations (Recuero et al. 2010; Alcaraz maturity to ascertain the impacts of environmental

et al. 2015; Contreras et al. 2009), such that the species conditions is not conducive to developing feasible and

is listed as Critically Endangered (Shaffer et al. 2004; — relevant conservation methods. By decreasing the time

Zambrano et al. 2010; Eisthen and Krause 2012). Few __ it takes to determine sex, management efforts can be

individuals exist outside of captivity with many axolotls — greatly expedited and both captive reproductive efforts

kept as exotic pets or maintained in laboratory colonies and field assessments can be improved..

where they are used as model organisms for biological One method of identifying sex of anuran amphibians

research (Contreras et al. 2009). is through hormone monitoring using urinary and fecal

Captive breeding and reintroduction programs are — steroid hormonal immunoassays (Germano et al. 2009,

being used to address dwindling wild populations and 2012; Hogan et al. 2013; Narayan 2013; Graham et al.

Correspondence. “chester_figiel@fws.gov

Amphib. Reptile Conserv. 71 February 2022 | Volume 16 | Number 1 | e302

Potential sex determination in Axolotls

2016). These studies infer that these endocrinological

methods could be implemented for identifying sex

in salamanders and caecilians. To that end, our

objectives in this study are to determine whether

hormone measurements are reliable methods for

identifying sex in salamanders. We investigated steroid

hormone metabolites in three age classes of axolotls,

A. mexicanum (Shaw and Nodder 1798), (classified

as juvenile, sub-adults, and adults) with the goal of

developing protocols for quantifying sexual hormones.

Previous methods for quantifying steroid metabolites

in axolotls required highly invasive procedures that

resulted in termination of the experimental subjects

(Eisthen and Krause 2012). Our goals were to develop a

procedure which posed minimal risk for the subjects that

could reliably quantify steroid hormone concentrations

in the three age classes, as defined above. Prior to this

study, we attempted to develop methods for extracting

urinary and fecal samples from axolotls. However,

we were unable to consistently collect adequate

volumes of urine and fecal matter which could reliably

quantify steroid metabolites so this study focuses on

tissue samples. Second, we explored the possibility

of differentiating between females and males by

examining differences in the concentrations of steroid

hormone metabolites (testosterone and 17 -estradiol

[hereafter estradiol]) and calculating differences in the

ratio of the concentrations of testosterone to estradiol.

Both objectives are an attempt to develop a foundation

for verifying sex of axolotls at an earlier age class.

Verifying the sex of juvenile axolotls, a surrogate for

other salamanders (especially Ambystoma species), may

aid in the management of various species of concern.

Materials and Methods

Animals

We used 13 reproductively mature adult A. mexicanum

that were maintained at the Warm Springs Fish

Technology Center in Warm Springs, Georgia, USA.

Of these, five were verified male subjects (snout-vent

length [SVL] = 145.7 mm + 10.0 mm), and eight were

verified female subjects (snout-vent length = 138.0 mm

+99 mm). We obtained 16 A. mexicanum of unknown

sex from the Ambystoma Genetic Stock Center (AGSC)

at the University of Kentucky and classified these either

as sub adults (SVL= 115.0 mm+3.9 mm; approximately

8 to 10 months old) or juveniles (SVL = 99.9 mm +

2.2 mm; approximately 5 to 6 months old). From these

individuals, we chose six sub-adults and six juveniles

for this study. The adult females, adult males, and sub-

adults were housed in individual tanks, while juveniles

were housed in tanks with other individuals which were

of similar size and age. All tanks were supplied with

a Steady flow of fresh spring water at a rate of ~0.5 L/

min. The axolotls were fed a standard pellet diet three

to four days per week (three pellets twice a day when

Amphib. Reptile Conserv.

fed). Food pellets, “soft-moist” pellets (5 mm in size)

were made by Rangen, Inc. and were purchased from

the AGSC.

Tissue Collection and Steroid Tissue Extraction

Toe clips provided tissue samples from eight adult female

subjects, five adult male subjects, and six sub-adults of

unconfirmed sex and six juveniles of unconfirmed sex.

Prior to sample collection, the animals were transferred

to buckets containing an anesthetic bath prepared with the

standard protocol for anamniote vertebrate sedation using

tricaine mesylate (MS-222). Each bath was prepared at

room temperature with 200 mg/L MS-222 in fresh tap

water. The pH of each bath was neutralized using NaOH,

which has previously been found to increase anesthetic

efficacy and reduce stress in amphibians (Robinson and

Scadding 1983). The bath was aerated with a solitary

pump while organisms occupied the bath. Toe clips

are a standard marking method for amphibian subjects

in ecological studies and have not been shown to have

a significant detrimental effect on other Ambystoma

species (Otto and Scott 1999). Based on the extensive

research on limb regeneration in A. mexicanum, it was

assumed that when executed with extreme care, toe clips

would not have a damaging effect on this species. Steroid

hormones are more likely to be stored in the bone than

the skin, and to maximize the amount of steroid hormone

that would be found in each sample, the middle digit of

the right forelimb was clipped on every subject. This is

the longest digit with the most bone, so it 1s assumed this

is the most efficient method of obtaining bone samples

while minimizing risk to the organism.

Samples were processed using the protocol developed

by Arbor Assays for steroid tissue extraction but modified

for small sample volumes. Samples were dried at 70

°C for four hours. Samples were weighed periodically

until constant weight was reached, and they were

completely dry. Samples were homogenized by mortar

and pestle and 1.0 g of each sample was transferred

to new microcentrifuge tubes. To each tube, 10 ml of

acetonitrile was added, and then the tube was sealed, and

vortexed thoroughly. To determine extraction efficiency,

one randomly chosen sample was split into two tubes and

10 uL of either 100,000 pg/mL estradiol or 200,000 pg/

mL testosterone was added in addition to acetonitrile.

All samples were centrifuged at 4,700 rpm at 4 °C for

15 minutes The supernatants of the centrifuged samples

were transferred to clean 15 mL conical tubes and frozen

at 4 °C overnight. The samples were gradually thawed

over two hours. Once thawed, solvents were extracted

using organic phase separations. To each sample, 30

ml of hexane was added and the sample was vortexed

for two minutes. The solution was transferred to a

separatory funnel, and the supernatant was transferred to

a clean tube while the top layer of hexane was discarded.

The supernatant which still contained acetonitrile was

evaporated to dryness in Speed Vac over the course of two

hours. The dried, extracted samples were reconstituted

using 100 uL of ethanol and then diluted with 450 uL of

Assay Buffer provided in the enzyme immunoassay kit

from Arbor Assays (Ann Arbor, Michigan, USA).

February 2022 | Volume 16 | Number 1 | e302

Veith and Figiel

Hormone Enzyme Immunoassays and Analysis

The reconstituted tissue samples were immediately run

on DetectX Testosterone ELISA kit (Cat. # K032-H1,

Arbor Assays, Ann Arbor, Michigan, USA) and 178

Estradiol ELISA kit (Cat. # KO30-H1, Arbor Assays,

Ann Arbor, Michigan, USA) based on instructions

provided in the kit. Serially diluted samples were

run alongside samples on each plate. All samples

and standards were run in duplicate. Plates were read

at an absorbance of 450 nm on a microplate reader.

Concentrations were calculated from a standard curve

using 4PLC fitting software provided by the free

online software MyAssays, Ltd. For both enzyme

immunoassays, one sample was extracted twice with a

standard concentration of steroid hormone added to one

portion of the sample to test extraction efficiency. Blank

wells were run on each plate as a control. Concentrations

that fell outside the range of the standard curve were

excluded from the analysis.

Data Analysis

Statistical analyses were conducted using Minitab®

Statistical Software. Testosterone and_ estradiol

concentrations were compared between adult males

and adult females using Student’s t-tests. The ratios

of testosterone:estradiol across categories were also

compared using Student’s t-tests. Extraction efficiency

was calculated according to each steroid metabolite kit.

Results

Two of the 27 samples were not processed due to low

sample weight after drying. The extraction efficiencies

were 46.3% for estradiol extraction and 6.7% for

testosterone extraction. The limit of detection for the

testosterone kit was 0.031 ng/mL and the limit of

detection for the estradiol kit was 0.027 ng/mL. All

samples read were above these limits. The regression

slopes were parallel to the standard curve for both

estradiol (R? = 0.998) and testosterone (R? = 0.997).

The intraassay coefficients of variation for estradiol and

testosterone were 11.4% and 34.9%, respectively. To

minimize variation in testosterone, it is suggested that

extraction efficiency be maximized and an increase in

serial dilutions be implemented.

Estradiol concentrations for females varied from 0.69

to 1.47 ng/mL, with a mean (+S.D.) of 1.56+0.23 ng/mL,

while estradiol concentrations for males ranged between

0.50 and 1.31 ng/mL, with a mean of 0.88 + 0.38 ng/

mL. Estradiol for subadults was between 0.49 and 1.27

ng/mL, with a mean of 0.78 + 1.12 ng/mL, whereas for

juveniles it was from 0.66 to 1.68 ng/mL, with a mean of

1.05 + 1.57 ng/mL. Estradiol concentrations were higher

than testosterone concentrations (Table 1), however

there was no significant difference in estradiol between

females and males (t [11] = 0.89, p = 0.3881).

Testosterone concentrations for females were between

0.15 and 0.63 ng/mL, with a mean of 0.359 + 0.18 ng/

mL, while testosterone concentrations for males ranged

0.07 to 0.28 ng/mL, with a mean of 0.17 + 0.09 ng/

Amphib. Reptile Conserv.

mL, and differences in testosterone concentrations

between the sexes were significantly different at the p =

0.0569 level (t [10] = 1.81). The range of testosterone

concentrations for subadults was 0.17 with a mean of

0.66 ng/mL, 0.30 + 0.75 ng/mL, and 0.15 with a mean

of 0.45 ng/mL, 0.24 + .05 ng/mL for juveniles. The ratio

of testosterone:estradiol concentrations did not differ

significantly between adult females and adult males (t

[10] = 1.81, p=0.1112).

Discussion

This study establishes a viable protocol for collecting

tissue samples that can be analyzed for steroid hormone

metabolite content. In addition, our findings validate the

modified steroid extraction protocol developed for this

study as both testosterone and estradiol were present in

detectable concentrations in the collected tissue samples.

The presence of these hormones in earlier developmental

stages was also confirmed, providing the prospect that

hormone changes can be tracked over the course of sexual

development. All estradiol concentrations for females

fell within previously reported ranges (0.2—2.8 ng/mL)

(Eisthen and Krause 2012), while testosterone levels in

males were lower than previously reported (0.2 to 0.8

ng/mL, Eisthen and Krause (2012)) potentially resulting

from the lower testosterone extraction efficiency (6.7%)

(Jacobs and Kuhn 1988).

It may be possible to determine sex based solely on

steroid hormone concentrations. Female axolotls had

over twice as much testosterone than males (on average),

however ranges of this hormone overlapped between

the sexes, so these results must be viewed with caution.

Future studies with a larger sample size and with methods

that optimize extraction efficiency would potentially

yield discernable differences in concentrations.

Previous studies on axolotls required dissection of

the organism or invasive techniques which resulted in

the termination of the subject to measure hormone levels

(e.g., Eisthen and Krause 2012). In this study, all subjects

survived the toe clip procedure, and no significant

detriment occurred in any animals from the loss of the

digit. This suggests that this procedure might be both

a reliable and feasible method for steroid hormone

measurement.

This study can serve as a basis for research into

the environmental sex determination interests of the

Species, and consequently the concern for alterations

of sex ratios in amphibian populations that might result

from rearing/growth in different water temperatures.

Axolotls are known to have a genetic sex determination

system, referred to as the ZZ/ZW system, with females

being the heterogametic sex (Armstrong and Malacinski

1989). Research by Bodney (1982) determined that

increased levels of estradiol benzoate in water would

induce sexual role reversal in male axolotls. Humphrey

(1948) examined the functional role reversal of adult

female axolotls due to the implementation of male sex

structures. Further, Eisthen and Krause (2012) found

that several environmental factors influenced estradiol

levels in axolotl females but not in males. These studies

show that axolotls have a flexible sexual system,

February 2022 | Volume 16 | Number 1 | e302

Potential sex determination in Axolotls

Table 1. Table of raw data for the estradiol (E) and testosterone (T) concentrations (ng/mL) of all tissue culture samples that were

processed and analyzed via enzyme immunoassay as well as the ratios of the concentrations (T:E). Concentration marked with

double asterisk (**) indicates the sample was out of range of the standard curve.

Subject Female Male

E T T:E E T

1 0.857 0.373 0.435 1.242 0.285

2 1.466 + n/a 1.310 0.086

3 0.966 0.633 0.655 0.557 0.206

4 FO125 “01559 “105525 5015 90.071

5 1.039 0.151 0.145 0.803 0.208

6 1143 0.251 0.241 - .

7 1.176 0.337 0.287 - -

8 0.693 0.215 0.310 - -

suggesting the determination of sex can be influenced by

environmental conditions. Hopefully, our research can

provide a foundation for the understanding of hormones

and sexual development and assist future investigations

of how climate changes influence sexual determination

and sex role reversal in A. mexicanum.

Acknowledgements.-We thank Emily Brown and the

Kubanek Lab at the Georgia Institute of Technology for

providing lab space, steroid extraction materials, and

guidance on the protocol. Additionally, we thank Yael

Toporek and the DiChristina Lab at Georgia Institute

of Technology for use of the microplate reader. We are

grateful to Jaclyn Zelko and William Wayman, both from

the Warm Springs Fish Technology Center, US Fish and

Wildlife Service, for their assistance with gathering

materials and with sampling collections, and Brian

Hickson at the Warm Springs Fish Health Center, US

Fish and Wildlife Service, for help in the development

of protocols. We thank Randal Voss, Laura Muzinic, and

Chris Muzinic at the Ambystoma Genetic Stock Center

of the University of Kentucky and the National Institutes

of Health grant that supports Ambystoma research

resource development: R240D010435. Any use of

a trade name or proprietary product does not imply

endorsement by the U.S. Government. The findings and

conclusions in this article are those of the authors and

do not necessarily represent the views of the U.S. Fish

and Wildlife Service. Links to commercial sites are in

no way an endorsement of any product or service of a

vendor.

Literature Cited

Alcaraz G, Lopez-Portela X, Robles-Mendoza C. 2015.

Response of a native endangered axolotl, Ambystoma

mexicanum (Amphibia), to exotic fish predator.

Hydrobiologia 753(1): 73-80.

Armstrong JB, Malacinski GM. 1989. Developmental

Biology of the Axolotl. Oxford University Press. New

York, New York, USA. 320 p.

Bodney SA. 1982. Towards the development of a

practical method for reversing the sex (male to

female) of large numbers of Ambystoma mexicanum.

Axolotl Newsletter 11:12-18.

Amphib. Reptile Conserv.

Subadult Juvenile

T:E E T T:E E T T:E

0.229 0.586 0.315 0537 0.738 0.225 0.304

0.066 0.781 0.167 0214 0.168 0189 0.113

0.370 0.690 0.259 0.375 1326 0-163. —.0:123

0.143 0.491 0.206 0418 0.102 0149 0.145

0.259 1272 0.659 0.518 0.905 0.453 0.501

0.862 0.205 0.237 0.659 0.264 0.400

Contreras V, Martinez-Meyer E, Valiente E, Zambrano

L. 2009. Recent decline and potential distribution in

the last remnant area of the microendemic Mexican

axolotl (Ambystoma mexicanum). Biological

Conservation 142(12): 2,881—2,885.

Eisthen HL, Krause BC. 2012. Ambiguities in

the relationship between gonadal steroids and

reproduction in axolotls (Ambystoma mexicanum).

General and Comparative Endocrinology 176(3).:

472-480.

Germano JM, Molinia FC, Bishop PJ, Cree A. 2009.

Urinary hormone analysis assists reproductive

monitoring and sex identification of bell frogs (Litoria

raniformis). Theriogenology 72(5): 663-671.

Germano JM, Molinia FC, Bishop PJ, Bell BD, Cree A.

2012. Urinary hormone metabolites identify sex and

imply unexpected winter breeding in an endangered,

subterranean-nesting frog. General and Comparative

Endocrinology 175(3): 464-472.

Humphrey RR. 1948. Reversal of sex in females of

genotype WW in the axolotl (Siredon or Ambystoma

mexicanum) and its bearing upon the role of the Z

chromosomes in the development of the testis. The

Journal of Experimental Zoology 109(2): 171-185.

Jacobs GFM, Kuhn ER. 1988. Luteinizing hormone-

releasing hormone induces thyroxine release together

with testosterone in the neotenic axolotl Ambystoma

mexicanum. General and Comparative Endocrinology

71(3): 502-505.

Narayan EJ. 2013. Non-invasive reproductive and stress

endocrinology in amphibian conservation physiology.

Conservation Physiology 1(1): cot011.

Ott J, Scott D. 1999. Effects of toe-clipping and PIT-

tagging on growth and survival in metamorphic

Ambystoma opacum. Journal of Herpetology 33(2):

344-348.

Recuero E, Cruzado-Cortes J, Parra-Olea G, Zamudio

KR. 2010. Urban aquatic habitats and conservation

of highly endangered species: the case of Ambystoma

mexicanum (Caudata, Ambystomatidae). Annales

Zoologici Fennici 47(4): 223-239.

Robinson ME, Scadding SR. 1983. The effect of pH on

tricaine methanesulfonate induced anaesthesia of the

newt Notophthalmus viridescens. Canadian Journal

of Zoology 61(3): 531-533.

February 2022 | Volume 16 | Number 1 | e302

Veith and Figiel

Shaffer B, Flores-Villela O, Parra Olea G, Wake D. Zambrano L, Mosig Reidl P, McKay J, Griffiths R, Shaffer

2004. Ambystoma mexicanum. The IUCN Red List B, Flores-Villela O. 2010. Ambystoma mexicanum.

of Threatened Species 2004: e.T1095A3229557. The IUCN Red List of Threatened Species 2010:

e.T1095A3229615.

Amphib. Reptile Conserv.

Isabel Veith received her B.S. in Biology with a Certificate in Environmental Science from the

Georgia Institute of Technology (Atlanta, Georgia, USA). The inception of this research project

began during Isabel’s time as an intern for the US Fish and Wildlife Service and gradually

transformed into her Senior Honor’s Thesis during her final year at Georgia Tech. Currently she

is living and working in Colorado Springs, Colorado, USA.

Chester R. Figiel, Jr. is a Supervisory Fish Biologist with the U.S. Fish and Wildlife Service

stationed at the Warm Springs Fish Technology Center in Warm Springs, Georgia, USA. He

received B.S. and MLS. degrees (in Biology) from Memphis State University, Tennessee, USA,

and a Ph.D. from the Biology Department at the University of Mississippi, Oxford, Mississippi,

USA. His current interests include amphibian reproductive biology, the influence of invasive

crayfish on native crayfishes, and the distribution of crayfish diseases in the southeastern United

States.

75 February 2022 | Volume 16 | Number 1 | e302

Official journal website:

amphibian-reptile-conservation.org

Amphibian & Reptile Conservation

16(1) [General Section]: 76-85 (e303).

Climate change and the fate of endemic Beysehir Frog,

Pelophylax caralitanus

‘Akin Kirag, 7Muge Gidis, *Ahmet Mert, and **Eyup Baskale

‘Canakkale Onsekiz Mart University, Canakkale Technical Sciences Vocational College, Canakkale, TURKEY *Kiitahva Dumlupinar University,

Faculty of Arts and Science, Department of Biochemistry, Kiitahya, TURKEY ?Isparta University of Applied Sciences, Faculty of Forestry, Isparta,

TURKEY *Pamukkale University, Faculty of Science and Arts, Department of Biology, Denizli, TURKEY

Abstract.—Global warming and the decline in precipitation threaten wetlands worldwide, and lakes in some

regions are in the process of drying. Amphibians, since they are water-dependent, will be the creatures

most affected by the rapid habitat losses due to climate change. Especially for amphibian species which are

endemic, the situation will be more serious in terms of its impact on biodiversity. Therefore, in this study, we

determined the climate characteristics specific to the habitats of an endemic amphibian species, Pelophylax

caralitanus. According to the Representative Concentration Pathways (RCP) climate change scenarios of the

ICPP, we analyzed whether the climatic characteristics specific to these habitats will exist in 2050 and 2070

under the criteria of RCP 2.6, RCP 4.5, RCP 6.0, and RCP 8.5. The results are quite alarming for Pelophylax

caralitanus. According to the RCP climate change scenarios, the climatic conditions in the present habitats of

this endemic amphibian species will not remain stable in that the potential habitats in Southwestern Anatolia

will be dramatically reduced and the appropriate habitats of P. caralitanus around the Turkish Lake District will

completely disappear, while some new potential habitats will emerge in the Northwest Aegean region of Turkey.

Keywords. Amphibian, climate change scenarios, habitat loss, MaxEnt, RCP, Representative Concentration Pathways,

Turkey

Citation: Kirag A, Gidis M, Mert A, Baskale E. 2022. Climate change and the fate of endemic Beysehir Frog, Pelophylax caralitanus. Amphibian &

Reptile Conservation 16(1) [General Section]: 76—85 (e303).

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.

Accepted: 7 November 2020; Published: 31 March 2022

Introduction

The Beysehir Frog, Pelophylax caralitanus, is an

endemic species of Turkey which is distributed across the

Mediterranean region of Turkey called the Turkish Lake

District (Arikan et al. 1994; Ayaz et al. 2006; Baskale et

al. 2017; Dusen et al. 2004; Kaya et al. 2002). According

to the literature on the distribution of P. caralitanus, it

inhabits permanent wetlands with rich aquatic vegetation,

including permanent ponds, rain pools, streams, rivers,

and irrigation channels. Pelophylax caralitanus prefers

abundant vegetation around lakes and ponds as habitats

(Baskale and Capar 2016). In particular, emergent

vegetation present in the water and the presence of

bushes and weeds around the water body constitute an

ideal area for the release of eggs and seeking shelter from

predators. Pelophylax caralitanus is listed by IUCN as

Near Threatened (NT) because of ongoing threats from

habitat loss and overexploitation (Oz et al. 2009). Since

it has a relatively wide distribution and presumed large

populations, it is unlikely to experience declines that are

Correspondence. “ebaskale@pau.edu.tr

Amphib. Reptile Conserv.

rapid enough to qualify it for listing in a higher Red List

category.

Climate change is one of the greatest problems

threatening biodiversity and ecosystems in the 21"

century, leading to the extinction of many species

(Sinervo et al. 2010; Walther et al. 2002). The habitat

preferences of amphibians are closely related to

temperature, precipitation, and wetlands, so amphibians

are one of the most sensitive animal groups to changes in

climatic conditions (Enriquez-Urzelai et al. 2019; Ortiz-

Yusty et al. 2013). Temperature was found to be positively

correlated with the detection probability of P. caralitanus

(Baskale and Capar 2016). Wetland ecosystems, in which

amphibians complete their life cycles, are among the

areas most affected by climate change, which creates

inhospitable conditions for amphibian reproduction

(Desta et al. 2012; Hopkins 2007). According to the

literature, the losses of amphibian habitat will be

high due to the increase in annual temperatures in the

Mediterranean basin, and it is predicted that there will be

new gains in the north (Araujo et al. 2006). In addition

March 2022 | Volume 16 | Number 1 | e222

Kirag et al.

to the difficulties amphibians face from possible climatic

changes, the pathogens that cause amphibian deaths,

such as Batrachochytrium dendrobatidis in the southern

part of the world, have been detected with increasing

frequency in the north due to climate change (Berger et

al. 1998; Cohen et al. 2019; Erismis et al. 2014).

The IPCC (Intergovernmental Panel on Climate

Change), which has been investigating climate change

for 30 years, presented new climate change scenarios

under the name of “Representative Concentration

Pathways” (RCP) in its Fifth Assessment Report (IPCC

2014). These scenarios estimate that global warming

will change atmospheric CO, concentrations by 2100 to

levels of 1,370 ppm for the RCP 8.5 scenario, 850 ppm

for RCP 6.0, 650 ppm for RCP 4.5, and 490 ppm for

RCP 2.6. These scenarios predict that temperatures in the

world may increase by anywhere from 1.5 °C to 5.8 °C

(IPCC 2014).

It is important to know the climatic requirements of a

species in order to predict how that species will respond

to climate change. The MaxEnt approach and software

have been widely used in recent years to determine the

current climatic demands of a species and to predict

changes in that species’ habitats in the future (Hendrick

and McGarvey 2019; Kirag and Mert 2019; Untalan et al.

2019; Zhang et al. 2018). MaxEnt gives more accurate

results with less data in smaller areas compared to

25°0'0"E 30°0'0"E

40°0'0"N

35°0'0"N

Fig. 1. Turkish Lake District and species presence data.

Amphib. Reptile Conserv.

35°0'0"E

other methods, e.g, DOMAIN, BIOCLIM, and GARP

(Hernandez et al. 2006; Wisz et al. 2008). In addition,

MaxEnt allows categorized and continuous data to be

processed together (Phillips et al. 2006; Phillips and

Dudik 2008) and it creates habitat suitability maps in

addition to the outcome outputs (Elith et al. 2006, 2011;

Hernandez et al. 2006, 2008).

The aim of this study is to investigate the ecological

niche of P. caralitanus and evaluate the consistency

and variations in the predicted potential distributions

of this endemic species in the Turkish Lake District in

the Southwest of Turkey under the IPCC’s four future

climate scenarios (RCP 2.6, RCP 4.5, RCP 6.0, and RCP

8.5). The results will help us to understand how this

endemic amphibian species will experience the effects of

climate change in relation to distributional shifts.

Materials and Methods

Study site. The Turkish Lake District (Fig. 1) is located

in Southwest Turkey and includes the western part of the

Taurus Mountains. This region contains many different

tectonic water bodies such as Beysehir Lake, Egirdir

Lake, Burdur Lake, Aksehir Lake, Salda Lake, and

Acig6l Lake. The Turkish Lake District is a transitional

region between the Mediterranean and continental

climates (Baskale et al. 2017; Dusen et al. 2004).

40°0'0"E 45°0'0"E

Beysehir Frog

Presence Data

March 2022 | Volume 16 | Number 1 | e222

Climate change and Pelophylax caralitanus

Data collection. Presence data for P caralitanus were

recorded from 89 sites in the Turkish Lake District area

based on our field studies performed between 2010

and 2016, as well as previously published papers (..e.,

Ayaz et al. 2006; Baskale and Capar 2016; Baskale et

al. 2017; Dusen et al. 2004; Kaya et al. 2002) (Fig. 1).

The sampling sites and their coordinates are given in

Appendix 1. Sampling from habitats during this study

was approved by the General Directorate of Nature

Conservation and Natural Parks in the Ministry of

Agriculture and Forestry, Turkey (protocol number

18.10.2010/61288) and the Animal Care and Use

Committee at Pamukkale University (protocol number

20.09.2010-PAUHDEK-2010/021). Individuals of the

target species were identified in their natural habitats

and were not exposed to stress. No measurements or

experiments were performed on the species.

Bioclimatic data. The current bioclimatic data (Biol

to Biol9, for years 1950-2000) were downloaded from

version 1.4 (30 arc-sec, or ~1 km) of the WorldClim

website (http://worldclim.org, Hijmans et al. 2005). The

varible definitions are given in Table 1. In Version 1.4,

HadGEM72-ES (30 arc-sec, or ~1 km) based data for 19

bioclimatic variables were available for the future (2050

and 2070) climate projections based on the RCP 2.6,

RCP 4.5, RCP 6.0, and RCP 8.5 scenarios. These data

sets are provided on a global scale, and were optimized

to the size of Turkey with the help of ArcGIS 10.2 and

converted to ASCII format.

Habitat suitability model. Due to the narrow study

area, high correlations can exist between bioclimatic

variables which may pose a problem during the analysis.

To eliminate the problem of multi-collinearity, we

applied Pearson Correlation Analysis (1? < 0.8) for the

19 bioclimatic variables. If a pair of variables has a

correlation coefficient of greater than 0.8, they were

considered to both represent a similar phenomenon,

so one of the pair of variables was excluded from the

analysis.

MaxEnt 3.4.1 software was used to estimate the

climatic conditions limiting the current distribution of

P. caralitanus and to compare the current situation with

future climate scenarios (Phillips etal. 2017). The presence

data of P. caralitanus in CSV format were entered into the

“Samples” section of MaxEnt 3.4.1 software. The current

bioclimatic data in ASCII format were entered into the

“Environmental layers” section. Then, 2050 and 2070

bioclimatic data files (RCP 2.6, RCP 4.5, RCP 6.0, and

RCP 8.5) were entered in the “Projection layers directory/

file” section to compare the future scenarios. MaxEnt

analysis was run using 90% of the records as training

data to build the model and the remaining 10% for testing

the model. Ten repetitions were made for each model so

that different training and test data would be processed in

the analysis. Any bioclimatic variables (Biol to Biol9)

that did not contribute to the model obtained as a result

of the analysis, were excluded in the next analysis.

Replicated run type Crossvalidate was selected for the

analysis along with the settings: maximum iterations

500, convergence threshold 0.00001, log file maxent.log,

and the default prevalence was set to 0.5. The analysis

process continued until the best model was obtained. We

used the “Area under the receiver operating characteristic

(ROC) curves” (AUC) values to determine the best

Table 1. Bioclimatic variables obtained from the WorldClim website (http://worldclim.org).

Code Bioclimatic variables Unit

Biol Annual Mean Temperature Ae

Bio2 Mean Diurnal Range (Mean of monthly (max temp - min temp)) io

Bio3 Isothermality ((Bio2/Bio7) * 100) unitless

Bio4 Temperature Seasonality (standard deviation * 100) CofV

Bio5 Max Temperature of Warmest Month °C

Bi06 Min Temperature of Coldest Month °C

Bio7 Temperature Annual Range (Bio5-Bio06) AS

Bio8 Mean Temperature of Wettest Quarter ae

Bio9 Mean Temperature of Driest Quarter Ae

Biol0 Mean Temperature of Warmest Quarter °C

Biol] Mean Temperature of Coldest Quarter "eC

Biol2 Annual Precipitation mm

Biol3 Precipitation of Wettest Month mm

Biol4 Precipitation of Driest Month mm

Biol5 Precipitation Seasonality (Coefficient of Variation) CofV

Biol6 Precipitation of Wettest Quarter mm

Biol7 Precipitation of Driest Quarter mm

Biol8 Precipitation of Warmest Quarter mm

Biol9 Precipitation of Coldest Quarter mm

Amphib. Reptile Conserv.

March 2022 | Volume 16 | Number 1 | e222

Kirag et al.

Cloglog Output

5000 6000 7000 ~—- 8000 9000 10000

Bio 4 (Temperature Seasonality)

0.9 = = =

Cloglog Output

200 250 300 350 400 450

Bio 7 (Temperature Annual Range)

Cloglog Output

-10 0 10 20°

Bio 14 (Precipitation of Driest Month)

30 50 60 70 80 90 100

359-200-150 100-30 0 50-100

Bio 6 (Min. Temperature of Coldest Month)

-100 -50 0 50 100 150 200 250

Bio 8 (Mean Temperature of Wettest Quarter)

-50 0 50 100 ~=—-150 200 250 300 350

Bio 14 (Precipitation of Driest Quarter)

Fig. 2. Variables with the highest contributions to the potential distribution of P. caralitanus according to MAXENT with the

standard errors in blue. The Y-axis indicates the probability of presence (based on the Cloglog, or complementary log-log transform,

values) and the X-axis shows the contribution of each variable.

model performance. An AUC value close to | indicates

that the model is in a perfect performance and a value

of 0.7 or higher indicates that the model is descriptive,

while a value of 0.5 indicates that the model does not

provide useful information (Phillips et al. 2006). Among

the models with the highest AUC values, the one with the

lowest standard deviation was selected as the best model.

Results

The results of the analysis indicate that the AUC value

of the training data is 0.965, while the test data AUC

value is 0.964, and the standard deviation is 0.012. We

Amphib. Reptile Conserv.

found that the climatic factors limiting the distribution of

P. caralitanus were: Bio4 (percent contribution: 22.3%,

Temperature Seasonality), Bio6 (16.4%, Min Temperature

of Coldest Month), Bio7 (17.9%, Temperature Annual

Range), Bio&8 (19.9%, Mean Temperature of Wettest

Quarter), Biol4 (15.5%, Precipitation of Driest Month),

and Biol7 (8%, Precipitation of Driest Quarter).

According to these results, P. caralitanus prefers habitats

with precipitation in the driest months, and those where

the average temperature of the coldest three months does

not fall below -5 °C, the seasonal temperature difference

is high, and the humid months are 0—5 °C (Fig. 2).

The habitats for P caralitanus were primarily in

March 2022 | Volume 16 | Number 1 | e222

Climate change and Pelophylax caralitanus

Mediterranean Sea

A Current Habitat Suitability Map

| Lakes and Sea

Value

* Species Data

4 Vaditerranean Sea

G RCP 6.0 2070 Climatic Habitat Suitability Map

— High: 0.965

Low: 0

H_ RP 8.5 2050 Climatic Habitat Suitability Map

. ‘i q

Mediterrancan Sea y”

C RCP 2.6 2070 Climatic Habitat Suitability Map

“¢f

sf. Mediterrancan Sea y

F RCP 6.0 2050 Climatic Habitat Suitability Map

Nedserranean Sea ra

| RCP 8.5 2070 Climatic Habitat Suitability Map

Fig. 3. Current climatic habitat suitability map (A) and the eight RCP climatic change scenario maps for P. caralitanus based on

RCP 2.6 (B—C), RCP 4.5 (D—E), RCP 6.0 (F—G), and RCP 8.5 (H-D) for either 2050 or 2070 as indicated.

Southwestern Anatolia, in the region between Central

Anatolia and Western Anatolia, and around the western

Taurus Mountains (Fig. 3A). A large part of the map

reflects the distribution areas of P. caralitanus reported

in the literature. Although its presence has not yet been

reported from the new potential areas, these areas are

climatically potential habitats.

If the RCP 2.6 scenario occurs, there will be habitat

losses in the Turkish Lake District until 2050, potential

habitats in the Southwest will persist, and new potentially

suitable bioclimatic habitats will emerge in the North

Aegean region (Fig. 3B). By 2070, the situation in the

Turkish Lake District will be slightly better than in 2050

and new potential habitats will be formed in a small area

in the inner Western Aegean region (Fig. 3C). In the case

of the RCP 4.5 scenario, suitable habitats in the Turkish

Lake District will almost disappear, new habitats will

appear in the Northern Aegean regions, and potential

habitats in the Southwest will persist (Fig. 3D-E). The

Amphib. Reptile Conserv.

RCP 6.0 scenario map shows that a situation similar to

the RCP 4.5 scenario in Fig. 3E will occur in 2050, but

it reveals that the situation will be worse in 2070 (Fig.

3F-G). According to the RCP 8.5 scenario, which is the

most dramatic result, potential habitats in Southwestern

Anatolia will be reduced but some will continue to exist,

and the appropriate habitats of P. caralitanus around the

Turkish Lake District will completely disappear. On the

other hand, new potential habitats not seen in the other

scenarios will emerge in the Northwest Aegean region

(Fig. 3H—-I).

Discussion

Recent studies have provided important information on

the extinctions of species due to climate change. Studies

have reported that there could be a 15—37% loss of species

by 2050 according to various climate change scenarios

(Thomas et al. 2004). Even in the most optimistic climate

March 2022 | Volume 16 | Number 1 | e222

Kirag et al.

change scenario, most of these losses will occur in

endemic species (Dirnboéck et al. 2011). In response to

climate change, many species will present compensating

mechanisms such as adaptation and migration; however,

the success of these mechanisms against climate change

will be related to the climate change velocity. In contrast

to species which have large areas of distribution,

endemic species distributed in a narrow region according

to special climatic and topographic conditions will not be

as successful, except in areas with a low climate change

velocity. In areas with a high climate change velocity,

endemism is either very low or does not exist. For this

reason low climate change velocity areas will be essential

refuges for many small-ranged species, such as endemic

species (Sandel et al. 2011).

The current habitat suitability map (Fig. 3A) shows

the distribution of P. caralitanus mainly in the Turkish

Lake District region and is compatible with species

observation data. It also indicates that there are suitable

habitats for this species in the north-western parts of the

Turkish Lake District (Fig. 3A), while recent studies have

found that the most north-western distribution area of the

species 1s Acig6l and Isikli Lakes (Baskale et al. 2017).

These lakes are located at the north-western border of

the Turkish Lake District. Although there are locations

with similar climatic conditions, the distribution of the

species in the northern part is geographically isolated due

to mountains.

In the past two decades, some lakes in the Turkish Lake

District have reduced water levels due to climatic changes

(Goncu et al. 2017; Kantarci 2008). Wetlands and lakes

are the essential elements of the habitats of frog species

such as P. caralitanus because they need water bodies to

complete their life cycle. A significant reduction of snow

in higher regions due to climate change, a reduction in

the water storage basins and earlier melting of snow

will have negative consequences for many wetlands in

Turkey (Yilmaz et al. 2019). Potential distribution maps

determined for P. caralitanus can help in prioritizing the

planning of future wetland use management around the

current populations, the discovery of new populations or

the identification of top priority areas.

According to RCP scenarios that were created here for

P. caralitanus, the climate change velocity will be high

for the next 30 to 50 years (Fig. 3B—I), and habitat loss

and fragmentation will be quite high in the distribution

area of the species. For the survival of endemic species,

we found RCP 2.6 to be the most optimistic scenario

among the climate scenarios. However, even in this

scenario, if P. caralitanus cannot migrate to the north-

western Turkish Lake District, the distribution of the

species will shrink in the next 30 years, and a very

narrow distribution area will host the species in 2070.

The IPCC, which is currently in its 6" assessment phase,

reported “Global warming of 1.5 °C” towards the end

of 2018, and emphasized that a temperature rise by 1.5

°C was better than 2 °C (IPCC 2018). According to our

Amphib. Reptile Conserv.

results, 1f the RCP 2.6 scenario is exceeded, most small-

scale wetlands will begin to dry in the next 30 years, and

irreversible changes may occur in the following years

relative to the current distribution map of P. caralitanus

(Fig. 3D-I). Although new locations with similar climate

zones occur in the north and north-west of the Turkish

Lake District, it would be very difficult for the frogs to

migrate from the original habitats to the new climatically

suitable habitats within the next 30 and 50 years because

of geographic isolation. Moreover, if the RCP 8.5 climate

scenario occurs, there will be no climatically suitable

habitats for P. caralitanus in the Turkish Lake District,

which we can categorize as a disaster.

We have shown that the MaxEnt model is a useful tool

for creating a predictive distribution map of P. caralitanus

in the Turkish Lake District, and it has allowed us

to determine putative environmental constraints and

successfully predict the species potential fate in the

coming years. According to our results, P. caralitanus

will move farther towards extinction in future, in the

face of such problems as an increase in temperature,

decrease in precipitation, loss of habitat, and reduction of

water bodies. In order to ensure the continuation of this

endemic species, the protection of habitat and the impact

of climate change on the species should be investigated

in detail with more environmentalist approaches.

Acknowledgements.—This_ research was supported

by Pamukkale University Scientific Research Projects

Unit- BAP (Project No: 2010BSP017). The permissions

for field work and handling of the frogs were issued by

the Animal Ethics Committee of Pamukkale University

(Pamukkale, Turkey) and the Ministry of Agriculture and

Forestry, General Directorate of Nature Conservation

and Natural Parks (Ankara, Turkey).

Literature Cited

Araujo MB, Thuiller W, Pearson RG. 2006. Climate

warming and the decline of amphibians and reptiles

in Europe. Journal of Biogeography 33: 1,712—1,728.

Arikan H, Ozeti N, Cevik I, Tosunoglu M. 1994.

Distribution of Rana ridibunda caralitana (Anura:

Ranidae) in the Lakes District. Turkish Journal of

Zoology 18: 141-145.

Ayaz D, Tok CV, Mermer A, Tosunoglu M, Afsar M,

Cicek K. 2006. A new locality for Rana ridibunda

caralitana Arikan, 1988 (Anura: Ranidae) in the

Central Anatolia. E.U. Journal of Fisheries and

Aquatic Sciences 23: 181-183.

Baskale E, Capar D. 2016. Detection probability and

habitat selection of the Beysehir Frog, Pelophylax

caralitanus (Arikan 1988), in southwestern Anatolia,

Turkey. Russian Journal of Herpetology 23: 205-214.

Baskale E, Sozbilen D, Polat F. 2017. Population ecology

and distribution of Pelophylax caralitanus (Arikan,

1988), in the Lakes District, southwestern Anatolia,

March 2022 | Volume 16 | Number 1 | e222

Climate change and Pelophylax caralitanus

Turkey. Herpetozoa 29: 143-153.

Berger L, Speare R, Daszak P, Green DE, Cunningham

AA, Goggin CL, Slocombe R, Ragan MA, Hyatt

AD, McDonald KR. 1998. Chytridiomycosis causes

amphibian mortality associated with population

declines in the rain forests of Australia and Central

America. Proceedings of the National Academy of

Sciences of the United States of America 95: 9,031—

9,036.

Cohen JM, Civitello DJ, Venesky MD, McMahon TA,

Rohr JR. 2019. An interaction between climate change

and infectious disease drove widespread amphibian

declines. Global Change Biology 25: 927-937.

Desta H, Lemma B, Fetene A. 2012. Aspects of climate

change and its associated impacts on wetland

ecosystem functions: a review. Journal of American

Science 8: 582-596.

Dirnbock T, Essl F, Rabitsch W. 2011. Disproportional

risk for habitat loss of high-altitude endemic species

under climate change. Global Change Biology 17:

990-996.

Dusen S, Oz M, Tunc MR, Kumlutas Y, Durmus H.

2004. Three new localities for Rana bedriagae

caralitana Arikan, 1988 (Anura: Ranidae) in the West

Mediterranean Region. Turkish Journal of Zoology

28: 115-117.

Elith J, Philltps SJ, Hastie T, Dudik M, Chee YE, Yates

CJ. 2011. A statistical explanation of MaxEnt for

ecologists. Diversity and Distributions 17: 43-57.

Elith J, Graham CH, Anderson RP, Dudik M, Ferrier S,

Guisan A, Hijmans RJ, Huettmann F, Leathwick JR,

Lehmann A, et al. 2006. Novel methods improve the

prediction of species’ distributions from occurrence

data. Ecography 29: 129-151.

Enriquez-Urzelai U, Bernardo N, Moreno-Rueda G,

MontoriA, Llorente G. 2019. Are amphibians tracking

their climatic niches in response to climate warming?

A test with Iberian amphibians. Climatic Change 154:

289-301.

Erismis UC, Konuk M, Yoldas T, Agyar P, Yumuk

D, Korcan SE. 2014. Survey of Turkey’s endemic

amphibians for chytrid fungus Batrachochytrium

dendrobatidis. Diseases of Aquatic Organisms 111:

153-157.

Gonctt S, Albek EA, Albek M. 2017. Trend analysis

of Burdur, Egirdir, Sapanca, and Tuz Lake water

levels using nonparametric statistical methods.

Afyon Kocatepe University Journal of Science and

Engineering 17: 555-570.

Hendrick LR, McGarvey DJ. 2019. Climate change and

mountaintop-removal mining: a MaxEnt assessment

of the potential threat to West Virginian fishes.

Northeastern Naturalist 26: 499-522.

Hernandez P, Franke I, Herzog S, Pacheco V, Paniagua

L, Quintana H, Soto A, Swenson J, Tovar C, Valqui

T. 2008. Predicting species distributions in poorly-

studied landscapes. Biodiversity and Conservation

Amphib. Reptile Conserv.

17: 1,353—1,366.

Hernandez PA, Graham CH, Master LL, Albert DL. 2006.

The effect of sample size and species characteristics

on the performance of different species distribution

modeling methods. Ecography 29: 773-785.

Hiymans RJ, Cameron SE, Parra JL, Jones PG, Jarvis

A. 2005. Very high resolution interpolated climate

surfaces for global land areas. /nternational Journal

of Climatology 25: 1,965—1,978.

Hopkins WA. 2007. Amphibians as models for studying

environmental change. JLAR Journal 48: 270-277.

IPCC. 2018. Global Warming of 1.5 °C. An IPCC Special

Report on the Impacts of Global Warming of 1.5 °C.

United Nations, Intergovernmental Panel on Climate

Change, Geneva, Switzerland. Available: https://

www.ipcc.ch/sr15/ [Accessed: 1 May 2020].

IPCC. 2014. Climate Change 2013: The Physical Science

Basis. 5 Assessment Report (AR5). United Nations,

Intergovernmental Panel on Climate Change, Geneva,

Switzerland. Available: https://www.ipcc.ch/sr15/

[Accessed: 1 May 2020].

Kantarci M. 2008. Istinma-Kuraklasma Sirecinin Goller

Bolgesindeki Durumu ve Etkileri Uzerine Ekolojik

Bir DeSerlendirme. Tiirkiye Ormancilik Dergisi 9:

1-34.

Kaya U, Cevik IE, Erismis UC. 2002. New distributional

records for Rana bedriagae caralitana in Anatolia.

Turkish Journal of Zoology 26: 381-383.

Kirag A, Mert A. 2019. Will Danford’s Lizard become

extinct in the future? Polish Journal of Environmental

Studies 28: 1,741—1,748.

Ortiz-Yusty CE, Paez V, Zapata FA. 2013. Temperature

and precipitation as predictors of species richness

in northern Andean amphibians from Colombia.

Caldasia 35: 65-80.

Oz M, Kaska Y, Kumlutas Y, Kaya U, Avci A, Uziim N,

Yentyurt C, Akarsu F, Kasparek M. 2009. Pelophylax

caralitanus. The IUCN Red List of Threatened

Species 2009: e.T135806A4203649.

Phillips SJ, Anderson RP, Dudik M, Schapire RE, Blair

ME. 2017. Opening the black box: an open-source

release of MaxEnt. Ecography 40: 887-893.

Phillips SJ, Anderson RP, Schapire RE. 2006. Maximum

entropy modeling of species geographic distributions.

Ecological Modelling 190: 231-259.

Phillips SJ, Dudik M. 2008. Modeling of species

distributions with MaxEnt: new extensions and a

comprehensive evaluation. Ecography 31: 161-175.

Sandel B, Arge L, Dalsgaard B, Davies R, Gaston K,

Sutherland W, Svenning J-C. 2011. The influence of

Late Quaternary climate change velocity on species

endemism. Science 334: 660-664.

Sinervo B, Mendez-De-La-Cruz F, Miles DB, Heulin B,

Bastiaans E, Villagran-Santa Cruz M, Lara-Resendiz

R, Martinez-Méndez N, Calderon-Espinosa ML,

Meza-Lazaro RN. 2010. Erosion of lizard diversity

by climate change and altered thermal niches. Science

March 2022 | Volume 16 | Number 1 | e222

Kirag et al.

328: 894-899.

Thomas CD, Cameron A, Green RE, Bakkenes M,

Beaumont LJ, Collingham YC, Erasmus BF, De

Siqueira MF, Grainger A, Hannah L. 2004. Extinction

risk from climate change. Nature 427: 145-148.

Untalan M, Burgos D, Martinez K. 2019. Species

distribution modelling of two species endemic to

the Philippines to show the applicability of MaxEnt.

International Archives of the Photogrammetry,

Remote Sensing, and Spatial Information Sciences

42: 449-454.

Walther G-R, Post E, Convey P, Menzel A, Parmesan

C, Beebee TJ, Fromentin J-M, Hoegh-Guldberg

amphibians and reptiles.

O, Bairlein F. 2002. Ecological responses to recent

climate change. Nature 416: 389.

Wisz MS, Hiymans R, Li J, Peterson AT, Graham C,

Guisan A, Group NPSDW. 2008. Effects of sample

size on the performance of species distribution

models. Diversity and Distributions 14: 763-773.

Yilmaz YA, Aalstad K, Sen OL. 2019. Multiple remotely

sensed lines of evidence for a depleting seasonal

snowpack in the Near East. Remote Sensing 11: 483.

Zhang K, Yao L, Meng J, Tao J. 2018. MaxEnt modeling

for predicting the potential geographical distribution

of two peony species under climate change. Science

of the Total Environment 634: 1,326—1,334.

Akin Kirac is an Associate Professor in the Technical Science Vocational School at Canakkale

Onsekiz Mart University, Canakkale, Turkey. His primary research interests are mainly focused on

habitat suitability models and climate envelope models, and he has conducted research in the field

of species ecology and climate change. He has also authored or co-authored several articles in peer-

reviewed scientific journals on insects, amphibians, reptiles, and mammals.

Muge Gidis is an Associate Professor in the Department of Biochemistry at the University of

Dumlupinar in Kttahya, Turkey. She is interested in molecular ecology and the conservation biology

of reptiles and amphibians. Her recent research projects involve the comparative phylogeography of

Ahmet Mert is currently working as an Associate Professor in the Department of Wildlife Ecology

Amphib. Reptile Conserv.

and Management at Isparta University of Applied Sciences in Isparta, Turkey. His primary research

interests are mainly focused on modelling and mapping the habitat distribution of wildlife species

using geographic information systems. He is an expert in using new and advanced technologies

for wildlife species including mammals, birds, and reptiles. He combines and analyzes wildlife

species data through traditional field work and novel technologies to provide a deeper knowledge

and understanding of the factors that influence the distribution and abundance of wildlife species.

Furthermore, he has authored or co-authored several articles in peer-reviewed scientific journals.

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 a special

focus on the conservation of amphibians and reptiles living in Turkey. Eyup currently coordinates

several research projects focusing on threatened species in Turkey.

March 2022 | Volume 16 | Number 1 | e222

Appendix 1. The list of localities of Pelophylax caralitanus in this survey.

Site No

0 AN NHN Hn FB W NY

— —

= OS,

Climate change and Pelophylax caralitanus

Locality

Acig6l site 1/Denizli

Acig6l site 2/Denizli

Acig6l site 3/Denizli

Acigol site 4/Denizli

Acig6l site 5/Denizli

A$lasun site 1/Burdur

AS$lasun site 2/Burdur

A$lasun site 3/Burdur

Akgol site 1/Burdur

Akg6ol site 2/Burdur

Akg6ol site 3/Burdur

Aksu river/Antalya

Aksehir lake site 1/Konya

Aksehir lake site 2/Konya

Aksehir lake site 3/Konya

Belcegiz/Isparta

Beysehir site 1/Konya

Beysehir site 2 /Konya

Beysehir Site 3/Konya

Beysehir site 4/Konya

Beysehir site 5/Konya

Bucak/Denizli

Burdur lake site 1/Burdur

Burdur lake site 2/Burdur

Burdur lake site 3/Burdur

Derebucak site 1/Konya

Derebucak site 2/Konya

Eber lake site 1/Afyon

Eber lake site 2/Afyon

Eber lake site 3/Afyon

Eber lake site 4/Afyon

Esirdir lake site 1/Isparta

Esirdir lake site 2/Isparta

Esirdir lake site 3/Isparta

Egirdir lake site 4/Isparta

Egirdir lake site 5/Isparta

Egirdir lake site 6/Isparta

Fele/Isparta

Gedikli/Isparta

Gencek/Konya

Goékg6l/Denizli

Gokhoytik/Konya

Golctik site 1/Isparta

Golctik site 2/Isparta

Golctik site 3/Isparta

Amphib. Reptile Conserv.

GPS Coordinates

Latitute Longitute

37°46.375°N 29°50.680’°E

37°49.794"N 29°56.879°E

37°46.348’N 29°50.5817E

37°47.003’N 29°51.709°E

37°50.942’N 29°59.452°E

37°36.065’N 30°32.018°E

37°35.967°N 30°32.420°E

37°36.195’N 30°32.406°E

37°41.323’N 29°44.759E

37°40.268’N 29°44 .206°E

37°39.887°N 29°45.293°E

37°14.896’N 30°47.431E

38°26.834’N 31°21.99VE

38°32.596’N 31°22.835°E

38°31.314°N 31°19.875°E

37°57.680°N 31°19.188°E

37°58.268°N 31°25.984E

37°53.332’N 31°30.742’E

37°47.691°N 31°35.816’°E

37°39. 479°N 31°41.819°E

37°44 .304°N 31°41.056’E

38°15.035’N 29°51.8977E

37°41.411’N 30°03.452’E

37°38.995°N 30°03.628’°E

37°38.225°N 30°05.685°E

37°21.654°N 31°32.842’°E

37°25.667’°N 31°30.631’E

38°36.832’N 31°13.561E

38°39.140’N 31°14.51S°E

38°42.375°N 31°11.5017E

38°37.808’N 31°04.053°E

38°05.178’°N 30°56.857°7E

38°16.532’N 30°51.5817E

38°16.723’N 30°52.190°E

38°01.231’N 30°48.872°E

38°08.231’N 30°52.789°E

37°50.550’N 30°51.913°E

38°02.435’N 31°27. 156’E

37°53.334°N 31°20.392’°E

37°26.572’N 31°32.438°E

38°12.207°N 30°02.938°E

37°24.056’N 31°56.421’E

37°43.707'N 30°29.510’E

37°44.469°N 30°29.030’E

37°44.469°N 30°29.030°E

March 2022 | Volume 16 | Number 1 | e222

Altitude (m)

845

84]

840

839

840

1,024

1,023

1,024

994

992

994

103

954

1,127

1,132

1,126

1,136

1,128

1,126

822

910

859

865

1,261

1,281

971

970

920

921

1,234

1,127

1,354

821

1,021

1,387

1,346

1,347

Kirag et al.

Appendix 1 (continued). The list of localities of Pe/ophylax caralitanus in this survey.

Site No

Locality

Golhisar channel/Burdur

Golhisar dam/Burdur

Golhisar Lake/Burdur

Golyaka/Konya

Golytizt/Konya

Hufgla/Konya

Isikli lake site 1/Denizli

Isikli lake site 2/Denizli

Isikli lake site 3/Denizli

Isikli lake site 4/Denizli

Isikli lake site 5/Denizli

Isikli lake site 6/Denizli

Isikli/Denizli

Karakoy site 1/Burdur

Karakoy site 2/Burdur

Karamik site 1/Afyon

Karamik site 2/Afyon

Karamik site 3/Afyon

Karaoz/Antalya

Kavakkoy/Denizli

Kirkg6z site 1/Antalya

Kirkg6z site 2/Antalya

Kirkg6z site 3/Antalya

Kovada/Isparta

Kumluca site 1/Burdur

Kumluca site 2/Burdur

Salda lake site 1/Burdur

Salda lake site 2/Burdur

Salda lake site 3/Burdur

Salda lake site 4/Burdur

Salda lake site 5/Burdur

Saraykoy/Konya

Sarikaya/Isparta

Seydisehir site 1/Konya

Seydisehir site 2/Konya

Seydisehir site 3/Konya

Sug6zti/Isparta

Sula lake site 1/Konya

Sula lake site 2/Konya

Sula lake site 3/Konya

Tasagil/Konya

Tepearasi/Konya

Yalihtiyik/Konya

Yesilda&/Konya

Amphib. Reptile Conserv.

GPS Coordinates

Latitute Longitute

37°02.734°N 29°27.960°E

37°01.385’N 29°26.687°E

37°06.711°N 29°35.376°E

37°41.825’N 31°26.898°E

37°21.225’N 31°%53.425°E

37°28.112’N 31°34.050’°E

38°11.014-N 29°46.840°E

38°15.332’N 29°51.811VE

38°11.176°N 29°59. 55VE

38°14.403’N 29°57.245°E

38°16.443’N 29°54.415°E

38°12.368’N 30°01.774E

38°19.255’N 29°51.375°E

37°28.333’N 29°32.717E

37°28.561’N 29°32.385°E

38°27.915°N 30°52.449°E

38°23.005’N 30°45 .394VE

38°26.173’N 30°53.148°E

37°11.067°N 30°48 .640°E

38°09.254’°N 29°38.2717E

37°06.600’N 30°34.828°E

37°05.669’N 30°35.125°E

37°05.526’N 30°34.997°E

37°36.755°N 30°53.886°E

37°38.270°N 30°02.836’°E

37°38.518°N 30°03.275°E

37°31.391’N 29°38.159E

37°31.142’N 29°37.493°E

37°31.725°N 29°39. 41VE

37°31.133’N 29°37.405°E

37°31.499°N 29°36.912’E

37°19.247°N 32°07.011’E

37°56.404’°N 31°18.464E

37°28.588’N 31°48.769°E

37°28.519°N 31°49.462°E

37°27.094’°N 31°51.634¢E

38°14.227°N 31°17.231E

37°21.501’°N 31°59.413°E

37°18.965’N 32°06.145°E

37°21.232’N 31°55.516°E

37°22.702’N 31°53.321VE

37°28.233’N 31°37.85S°E

37°19.256’N 32°06.284E

37°35.087°N 31°30. 1SVE

March 2022 | Volume 16 | Number 1 | e222

Altitude (m)

1,022

1,068

947

1,126

1,099

1,370

816

819

821

829

996

991

1,004

811

304

307

302

906

860

1,158

1,161

1,145

1,160

1,168

1,102

1,127

1,135

1,131

1,110

1,226

1,222

1,096

1,098

1,101

1,387

1,101

1,125

Official journal website:

amphibian-reptile-conservation.org

Amphibian & Reptile Conservation

16(1) [General Section]: 86—93 (e304).

Modification of a Water Hyacinth sieve and description of

Hubbard rakes for sampling small aquatic salamanders

12.*Zachary C. Adcock, ‘Michelle E. Adcock, *Bruce E. Hall, and ‘Michael R.J. Forstner

'Texas State University, Department of Biology, 601 University Drive, San Marcos, Texas 78666, USA *Cambrian Environmental, 4422 Packsaddle

Pass Suite 204, Austin, Texas 78745, USA *Jasper Wyman & Son (Wyman’s), 7 Wyman Road, Milbridge, Maine 04658, USA

Abstract.—Jollyville Plateau Salamanders (Eurycea tonkawae) can be difficult to detect and capture in

submerged leaf litter packs, woody debris, and vegetation. Here we describe the modification of a Water

Hyacinth sieve and introduce three designs of Hubbard rakes to effectively sample these cover objects. Data

are reported on the captures of E. tonkawae using the sieve and all three rakes, as well as captures of E.

pterophila, E. naufragia, E. chisholmensis, and several co-occurring tadpoles, small fishes, and invertebrates.

The application and success of these tools are described in detail for various cover types, water depths, and

substrates.

Keywords. Amphibia, Caudata, central Texas, cover objects, Eurycea, monitoring

Citation: Adcock ZC, Adcock ME, Hall BE, Forstner MRJ. 2022. Modification of a Water Hyacinth sieve and description of Hubbard rakes for sampling

small aquatic salamanders. Amphibian & Reptile Conservation 16(1) [General Section]: 86—93 (e304).

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.

Accepted: 1 December 2020; Published: 28 February 2022

Introduction

Jollyville Plateau Salamanders (Eurycea tonkawae)

are small, fully-aquatic salamanders endemic to central

Texas, USA (Chippindale et al. 2000), which are listed

as Threatened by the U.S. Fish and Wildlife Service

(USFWS 2013). Bowles et al. (2006) and USFWS

(2013) considered submerged rocks and gravel to be

the preferred habitat for this taxon, but it has also been

documented from leaf litter, woody debris, and aquatic

vegetation (Bowles et al. 2006; Chippindale 2005; Davis

et al. 2001; O’Donnell et al. 2008). Submerged rocks

are easily surveyed by overturning them and visually

searching underneath for salamanders (Bowles et al.

2006; Pierce et al. 2010; Sweet 1977). In contrast, E.

tonkawae are difficult to detect in leaf litter packs, woody

debris, and vegetation because these cover objects can

be dense and often occur on silty substrate (Bowles et al.

2006; Davis et al. 2001). Bowles et al. (2006) recognized

that this difficulty may have caused underestimates of E.

tonkawae relative abundance in large leaf packs.

Previous researchers have surveyed for salamanders

in submerged, dense leaf litter, and vegetation with pipe

and box samplers, dip nets, and seines (Shaffer et al.

1994; Skelly and Richardson 2010), but these techniques

are difficult to apply in shallow water (less than 15

cm) and in areas with gravel or bedrock substrates that

Correspondence. *zca3@mxstate.edu

Amphib. Reptile Conserv.

characterize E. tonkawae habitat (Z.C. Adcock, pers.

obs.). Sweet (1977) collected central Texas Eurycea

in gravel substrates by shoveling the material onto a

wire-mesh screen suspended over a large tray. This

method could be applied to leaf litter, woody debris, and

vegetation, but it requires bulky gear that may be difficult

for one surveyor to use or to transport in the field.

O’ Donnell et al. (2008) reported catching EF. tonkawae by

sweeping leaf litter into a large net, but this method does

not allow for easy quantification of the surveyed area and

has limited applicability to other cover objects.

Passive and active traps, such as funnel traps, drift

nets, leaf litter bags, and mopheads, can capture Eurycea

salamanders in various aquatic covers (Devitt and Nissen

2018; Pauley and Little 1998; USFWS 2014; Waldron et

al. 2003; Willson and Dorcas 2003; Willson and Gibbons

2010). However, the animals which are captured can die

if passive traps are not checked frequently (Willson and

Gibbons 2010), and all trapping methods require several

subsequent site visits which may not always be practical.

Some traps can also result in a size-biased sample

(Luhring et al. 2016).

Here, we provide the details for a modification of the

Godley Water Hyacinth (Hichhornia crassipes) sieve

(Godley 1982) and describe the use of this sieve and

Hubbard rakes to sample E. tonkawae ina variety of cover

types, water depths, and substrates. We designed these

February 2022 | Volume 16 | Number 1 | e304

Adcock et al.

tools to be small so they would work efficiently within

the often-narrow spring runs and cave streams occupied

by these salamanders and to allow a single researcher to

easily carry and operate the equipment. Like the Godley

sieve, and its predecessor, the Goin dredge (Goin 1942)

our modified sieve and rakes sample a known area, thus

enabling estimates of salamander densities. Although we

designed these sampling devices to capture E. tonkawae,

we demonstrate they are also effective for other central

Texas Eurycea salamanders, as well as several co-

occurring vertebrates and invertebrates.

Materials and Methods

Sieve. Our modified sieve design required 1.25 m of

1.9-cm x 8.9-cm (standard l-inch x 4-inch) untreated

pine lumber, eight 3.8-cm (1.5-inch) galvanized corner

braces, two galvanized gate handles, a 91-cm x 2.1-m

(3-foot <x 7-foot) roll of fiberglass window screening, a

1.27-cm x 61-cm x 152-cm (0.5-inch x 2-foot < 5-foot)

roll of 19-gauge galvanized steel hardware cloth, a box

of 1.27-cm (0.5-inch) stainless steel staples, and a can

of spar urethane (Table 1). We cut the lumber into two

30-cm and two 32-cm lengths to form a box frame with

30 cm =x 30 cm interior dimensions (Fig. 1A), but we

note that the interior dimension should be adjusted to

accommodate the target taxa, sampling site, and project

Fig. 1. (A) Top, (B) side, and (C) bottom of a salamander sieve. Scale: 30 cm. Photos by Michelle Adcock.

goals. We attached the corner braces on the outside of

the frame to eliminate any sharp corners inside the

sieve that could harm captured animals (Fig. 1B). We

then sealed the frame with spar urethane and attached

the gate handles after the frame dried. The bottom was

constructed by attaching window screening which was

supported by hardware cloth to the bottom of the frame

with staples (Fig. 1C). Because of the small sieve size,

staples adequately supported the bottom, and a bottom

brace as described by Godley (1982) was not required.

The materials to construct one sieve cost about USD

$56.00. However, much of this cost was associated

with excess materials because the smallest amount

available for purchase exceeded the amount needed for

construction (see Table 1). The construction of additional

sieves from the excess material (up to 10 total) would

only require the purchase of more lumber, corner braces,

and gate handles for about USD $20.50 per sieve.

When the water was deep enough and floating cover

objects (e.g., woody debris, unrooted vegetation) were

present, the sieve could be positioned underneath the

material and lifted straight up, as described by Goin

(1942, 1943) and Godley (1980, 1982). In shallow

water and situations with rooted vegetation, we used a

large dustpan to scoop gravel, leaf litter, woody debris,

vegetation, and the inhabitants into the sieve (Fig. 2A).

Dustpans with similar dimensions to our modified sieve

Table 1. Cost of materials for the construction of one salamander sieve.

Material Quantity Total approximate cost (US dollars)

1.9-cm x 8.9-cm x 1.8-m (standard 1-inch x 4-inch x 6-foot) untreated pine 1 $3.50

lumber

3.8-cm (1.5-inch) galvanized corner brace 8 $7.00

Spar urethane* 1 can $10.00

Galvanized gate handle 2 $10.00

91-cm x 2.1-m (3-foot x 7-foot) roll of fiberglass window screening* 1 roll $7.00

1.27-cm x 61-cm x 152-cm (0.5-inch x 2-foot x 5-foot) roll of 19-gauge 1 $6.50

galvanized steel hardware cloth*

1.27-cm (0.5-inch) stainless steel staples* Box of 1,000 $12.00

Total Cost $56.00

*The quantity of these materials indicates the smallest amount available for purchase, not the quantity needed for construction of

a single sieve.

Amphib. Reptile Conserv.

February 2022 | Volume 16 | Number 1 | e304

Tools for for sampling small aquatic salamanders

Fig. 2. Salamander sieve demonstration. (A) Cover objects are scooped into the sieve using a dustpan and (B) carefully searched for

fauna to (C—D) reveal a salamander. Red arrows identify a Jollyville Plateau Salamander (Eurycea tonkawae) trapped in the sieve.

Photos by Madison Torres (A) and Zach Adcock (B—D).

design are available at most hardware stores for about

USD $7.00. Nets, strainers, or other scooping devices

can also be used to fill the sieve, but we chose a dustpan

in order to collect the sample in a single scoop, rather

than multiple small scoops which may cause animals

to flee before capture. We washed the collected cover

material with water to rinse away silt, then carefully

sorted through it to find salamanders and co-occurring

fauna (Fig. 2 B—C). Once inside the sieve, salamanders

are unlikely to escape (Fig. 2D), reducing the false

absences often associated with these difficult-to-sample

cover types (Bowles et al. 2006; Davis et al. 2001).

Hubbard Rakes. Aluminum rakes were constructed by

Hubbard Rakes in Jonesport, Maine, USA (http://www.

hubbardrakes.com), and they are custom designs that

combine aspects of their lowbush blueberry, cranberry, and

sea glass rakes. Each rake costs about USD $50.00 plus

shipping and handling. The interior dimensions (30 cm x

30 cm x 11.5 cm) match our modified sieve dimensions for

comparable density estimates. All rakes have a backend

(30 cm x 14 cm x 11.5 cm) that is enclosed on all sides

and serves as a receptacle for scooped material. We drilled

large drain holes and small holes for window screen

Amphib. Reptile Conserv.

attachment into the receptacle, and lined the rakes with

window screening to prevent fauna from escaping through

the teeth and drain holes (Fig. 3).

We designed three rakes that differ in the sampling

edge (1.e., flat edge, short teeth, and long teeth) to

accommodate different cover objects (Fig. 4). The flat-

edged rake 1s scooped through the cover objects, like the

dustpan, but as it is drawn through the water column, all

material and inhabitants are entrapped in the receptacle.

The short-toothed rake has ~6.5 cm teeth, and the long-

toothed rake has ~15 cm teeth. Both are designed to rake

through dense, rooted vegetation and comb any resident

fauna out of the vegetation and into the rake receptacle.

As with the sieve, salamanders are unlikely to escape

once inside the rakes (Fig. 5).

Sampling. To test the efficacy of these devices, F.

tonkawae were sampled at springs in the vicinity of Round

Rock and Cedar Park, Texas, USA, from 2014 through

2019. From July 2014 to August 2016, E. tonkawae

captures and survey effort were quantified using the sieve

and Hubbard rakes as well as standard visual encounter

surveys by searching under rocks (see Bendik et al. 2014;

Pierce et al. 2010). In subsequent years, the sieve and

February 2022 | Volume 16 | Number 1 | e304

Adcock et al.

fark _ ”

MM tay;

Fig. 3. (A) Top, (B) side, and (C) back of a Hubbard rake showing receptacle backend with drain holes and holes for window screen

attachment using zip ties. Scale: 30 cm. Photos by Michelle Adcock.

Hubbard rakes were used to survey for other species of

central Texas Eurycea salamanders (1.e., E. pterophila,

E. naufragia, and E. chisholmensis) at various springs in

Hays and Williamson counties, Texas, USA.

Results

From July 2014 to August 2016, 325 E. tonkawae were

captured using the sieve and Hubbard rakes, compared

to 342 E. tonkawae in rock surveys, which corresponded

to 0.53 salamanders per sieve/rake sample and 0.02

salamanders per searched rock. The sieve and Hubbard

rakes were used to capture E. tonkawae in submerged

gravel, leaf litter packs, small woody debris, silt, and

several types of vegetation (e.g., floating, aquatic, and

emergent). In addition, E. pterophila, E. naufragia, and

E. chisholmensis were caught in these same cover types

at their respective springs.

In addition to the targeted Eurycea, these tools

captured a number of co-occurring tadpoles, fishes, and

invertebrates. Bycatch included Blanchard’s Cricket

Frog (Acris blanchardi) tadpoles, Rio Grande Leopard

Frog (Lithobates [Rana] berlandieri) tadpoles, small

sunfish (Lepomis sp.), small bass (Micropterus sp.),

Western Mosquitofish (Gambusia affinis), Slough Darters

(Etheostoma gracile), crayfish (family Cambaridae),

dragonfly and damselfly larvae (order Odonata), mayfly

Amphib. Reptile Conserv.

Fig. 4. (A) Flat-edged, (B) short-toothed, and (C) long-toothed Hubbard rake designs. Scale: 30 cm. Photos by Michelle Adcock.

larvae (order Ephemeroptera), giant water bugs (family

Belostomatidae), beetles (order Coleoptera), snails

(order Gastropoda), hellgrammites (family Corydalidae),

annelid worms (subclasses Hirudinea and Oligochaeta),

and amphipods (order Amphipoda).

Discussion

Approximately 49% of FE. tonkawae were captured

using the sieve and Hubbard rakes, and the remaining

51% were caught in traditional rock searching surveys.

The frequency of salamander observations per rock (=

0.02) was comparable to those reported by Pierce et

al. (2010) for E. naufragia but substantially lower than

the salamander observations per sieve/rake sample (=

0.53). However, we acknowledge that these tools sample

a larger area than the average rock size. Our goal was

not to evaluate the best survey methodology or overall

tool, but to demonstrate that most cover objects can be

efficiently sampled with proper tool design and selection.

Any potential differences in salamander or faunal

captures among sampling tools would be more indicative

of differences in cover object availability and use (Z.C.

Adcock, unpub. data). Most importantly, our efforts

demonstrate that the sieve and Hubbard rakes effectively

capture central Texas Eurycea salamanders in cover

objects that have been previously described as difficult

aera

February 2022 | Volume 16 | Number 1 | e304

Tools for for sampling small aquatic salamanders

pm = 7

to reveal a salamander. Red arrow identifies a Jollyville Plateau Salamander (Eurycea tonkawae) trapped in the rake. Photos by

Zach Adcock.

to sample (Bowles et al. 2006; Davis et al. 2001) and

in cover objects the USFWS considers to be suboptimal

habitat (USFWS 2013).

Our modified sieve and dustpan combination worked

particularly well in shallow water, as cover objects

could be scooped without losing water and material over

the edges of the dustpan. The dustpan was effective at

scooping gravel, leaf litter, small woody debris, silt,

and unrooted or weakly rooted vegetation into the sieve

(Fig. 6). The sieve was also effective when floating

cover objects were present in deep water, as previously

described (Goin 1942, 1943; Godley 1980, 1982). We

_—. =

a ne

Fig. 6. Examples of Jollyville Plateau Salamander (Eurycea tonkawae) cover objects that are effectively sampled using the

salamander sieve and Hubbard rakes. (A) Submerged leaf litter and exposed roots, (B) submerged woody debris, (C) middle of

springrun, noting aquatic vegetation with weak roots, as well as the springrun edges which are shallow with emergent vegetation,

and (D) deep, aquatic vegetation with durable roots and stems. Photos by Zach Adcock.

Amphib. Reptile Conserv. 90 February 2022 | Volume 16 | Number 1 | e304

Adcock et al.

found the sieve and dustpan combination to be ineffective

for submerged cover objects in deep water (over 30 cm

deep) and in vegetation with durable stems and roots.

When scooping material in deeper water, cover objects

(and likely fauna) spilled over the sides of the dustpan as

it was brought up through the water column. Likewise,

the dustpan was inadequate at pushing through durable

roots or emergent vegetation, undoubtedly causing

salamanders to retreat undetected. These deficiencies

prompted our idea for combining the sieve and dustpan

into a single tool, the Hubbard rakes.

The Hubbard rakes were capable in all water depths

due to the enclosed backend receptacle. The flat-edged

rake was effective in scooping all cover types except for

vegetation with durable stems and roots, and we simply

used the toothed rakes in these situations. The short-

toothed rake performed well in aquatic vegetation and

in emergent vegetation along creek edges. In emergent

vegetation zones (Fig. 6C), the long-toothed rake often

hit hard substrate (e.g., soil along the bank) before the

vegetation encountered the receptacle edge, which

allowed fauna to escape through the teeth. However,

the long-toothed rake worked particularly well in large

patches of aquatic vegetation in water over 15 cm deep

(Fig. 6D). The toothed rakes did not perform well in

gravel, leaf litter, or woody debris because these smaller

items fell through the teeth during scooping. We note that

we never impaled salamanders with the rake teeth, and

researchers are unlikely to do so if the rakes are used in

a combing motion.

These tools also allowed us to sample exposed

vegetation roots in addition to stems and leaves. We

frequently captured Eurycea salamanders by placing

submerged, exposed root clumps in the sieve or rakes and

washing with water, by combing through roots with the

toothed rakes, and by scraping the bottom of dense root

mats and undercut stream banks with the flat-edged rake

or the wood edge of the sieve.

Central Texas Eurycea salamanders typically escape

predators (and researchers) by diving into the interstitial

spaces of the substrate. Sweet (1977) exploited this

behavior with his wire-screen method, allowing

salamanders to “escape” through the screen and into the

collection tray. Our modified sieve and Hubbard rakes

also exploit this behavior. Washing the sampled material

created enough disturbance to cause most entrapped

salamanders to retreat out of the cover objects and

to the bottom of the sieve or rakes. We note that adult

central Texas Eurycea were often easily noticed in the

sieve and rake as they actively sought shelter. Small

juvenile salamanders (less than 15 mm total length) were

typically less active and frequently remained motionless.

Therefore, we caution researchers to carefully search the

sampled material and the sampling devices for juveniles.

During the surveys, sampled gravel, leaf litter, woody

debris, silt, and similar cover objects were placed

back into the streams to minimize habitat impacts.

Amphib. Reptile Conserv.

The toothed rakes caused minor damage to aquatic

and emergent vegetation when combing through roots,

stems, and leaves. Using the sieve and rakes results in

destructive sampling for weakly rooted vegetation, but

we rarely noticed the sampling impacts in subsequent

survey events. The fast-growing Watercress (Nasturtium

officinale) constituted much of our sampled vegetation,

and a monthly survey timeframe allowed ample time for

regrowth. We suggest that researchers be cognizant of

potential oversampling by considering vegetation growth

rates and their planned survey timing.

We modified the Water Hyacinth sieve and designed the

Hubbard rakes to capture E. tonkawae, but they proved to

be effective for other central Texas Eurycea salamanders

and several co-occurring vertebrates and invertebrates.

The density of salamanders and co-occurring fauna can

be easily calculated by dividing the number of captures

by the number of samples multiplied by the size of the

sampling device. These tools are undoubtedly applicable

to a wide variety of small, aquatic salamanders, tadpoles,

fishes, and invertebrates if the appropriate device 1s

matched to the cover objects to be sampled.

Acknowledgements.—We thank Steve Godley for

recommending that we explore designs for a hybrid

sieve and scoop, Ike Hubbard for design input and

manufacturing the rakes, and field help from Texas

State University and Cambrian Environmental. Andrew

MacLaren, Steve Godley, and the reviewers provided

helpful comments on earlier drafts of the manuscript.

The Williamson County Conservation Foundation, Texas

Department of Transportation, and PulteGroup, Inc.

provided funding and site access. Additional site access

was provided by the Avant, Fowler, Lyda, and Swinbank

families. We conducted this work in compliance with

Texas State University Institutional Animal Care and Use

Committee (0417_0513_ 07), Texas Parks and Wildlife

Department (SPR-0102-191 and SPR-0319-056), and

the U.S. Fish and Wildlife Service (TE039544-1 and

TE37416B-0).

Literature Cited

Bendik NF, Sissel BN, Fields JR, O’ Donnell LJ, Sanders

MS. 2014. Effect of urbanization on abundance of

Jollyville Plateau Salamanders (Eurycea tonkawae).

Herpetological Conservation and Biology 9: 206-

222.

Bowles BD, Sanders MS, Hansen RS. 2006. Ecology of

the Jollyville Plateau Salamander. (Eurycea tonkawae:

Plethodontidae) with an assessment of the potential

effects of urbanization. Hydrobiologia 553: 111-120.

Chippindale PT. 2005. Eurycea tonkawae (Chippindale,

Price, Wiens, and Hillis, 2000) Jollyville Plateau

Salamander. Pp. 764-765 In: Amphibian Declines:

The Conservation Status of United States Species.

Editor, Lannoo M. University of California Press,

February 2022 | Volume 16 | Number 1 | e304

Tools for for sampling small aquatic salamanders

Berkeley, California, USA. 1,094 p.

Chippindale PT, Price AH, Wiens JJ, Hillis DM.

2000. Phylogenetic relationships and systematic

revision of central Texas hemidactyline plethodontid

salamanders. Herpetological Monographs 14: 1-80.

Davis B, Hansen R, McClintock NL, Peacock ED, Turner

M, Herrington C, Johns D, Chamberlain DA, Cerda

D, Pennington T, et al. 2001. Jollyville Plateau Water

Quality and Salamander Assessment. City of Austin

Watershed Protection Department, Austin, Texas,

USA. 381 p.

Devitt TJ, Nissen BD. 2018. New occurrence records

for Eurycea sosorum Chippindale, Price, and Hillis,

1993 (Caudata, Plethodontidae) in Travis and Hays

counties, Texas, USA. Check List 14: 297-301.

Godley JS. 1980. Foraging ecology of the Striped Swamp

Snake, Regina alleni, in southern Florida. Ecological

Monographs 50: 411-436.

Godley JS. 1982. Aquatic snakes in water hyacinth

communities. Pp. 281-282 In: CRC Handbook of

Census Methods for Terrestrial Vertebrates. Editor,

Davis DE. CRC Press, Boca Raton, Florida, USA.

397 p.

Goin CJ. 1942. A method for collecting the vertebrates

associated with water hyacinths. Copeia 1942: 183-

184.

Goin CJ. 1943. The lower vertebrate fauna of the water

hyacinth community in Northern Florida. Proceedings

of the Florida Academy of Sciences 6: 143-153.

Luhring TM, Connette GM, Schalk CM. 2016. Trap

characteristics and species morphology explain

size-biased sampling of two salamander species.

Amphibia-Reptilia 37. 79-89.

O’ Donnell L, Glusenkamp A, Herrington C, Schlaepfer

M, Turner M, Bendik N. 2008. Estimation of

Jollyville Plateau Salamander Populations using

Surface Counts and Mark-recapture. City of Austin

Watershed Protection Department, Austin, Texas,

USA. 110 p.

Pauley TK, Little M. 1998. A new technique to monitor

larval and juvenile salamanders in stream habitats.

Banisteria 12: 32—36.

Pierce BA, Christiansen JL, Ritzer AL, Jones TA.

2010. Ecology of Georgetown Salamanders

Amphib. Reptile Conserv.

(Eurycea naufragia) within the flow of a spring. The

Southwestern Naturalist 55: 291—297.

Shaffer HB, Alford RA, Woodward BD, Richards SJ,

Altig RG, Gascon C. 1994. Quantitative sampling of

amphibian larvae. Pp. 130-141 In: Measuring and

Monitoring Biological Diversity: Standard Methods

for Amphibians. Editors, Heyer WR, Donnelly MA,

McDiarmid RW, Hayek LC, Foster MS. Smithsonian

Institution Press, Washington, DC, USA. 364 p.

Skelly DK, Richardson JL. 2009. Larval sampling. Pp.

55-68 In: Amphibian Ecology and Conservation: a

Handbook of Techniques. Editor, Dodd CK. Oxford

University Press, Oxford, United Kingdom. 585 p.

Sweet SS. 1977. Natural metamorphosis in Eurycea

neotenes, and the generic allocation of the Texas

Eurycea (Amphibia: Plethodontidae). Herpetologica

33: 364-375.

U.S. Fish and Wildlife Service (USFWS). 2013.

Endangered and threatened wildlife and_ plants;

determination of endangered species status for the

Austin Blind Salamander and threatened species status

for the Jollyville Plateau Salamander throughout their

ranges; final rule. Federal Register 78(151): 51,278—

S1S326:

US. Fish and Wildlife Service (USFWS). 2014. United

States Fish and Wildlife Service Section 10(a)(1)

(A) Scientific Permit Requirements for Conducting

Georgetown, Jollyville Plateau, and _ Salado

Salamander Surveys. 18 p. Available: https://www.

fws.gov/guidance/sites/default/files/documents/118_

salamander _Survey_protocpro.pdf [Accessed: 3 April

2020).

Waldron JL, Dodd Jr CK, Corser JD. 2003. Leaf

litterbags: factors affecting capture of stream-dwelling

salamanders. Applied Herpetology 1: 23-36.

Willson JD, Dorcas ME. 2003. Quantitative sampling

of stream salamanders: comparison of dipnetting and

funnel trapping techniques. Herpetological Review

34: 128-129.

Willson JD, Gibbons JW. 2009. Drift fences, coverboards,

and other traps. Pp. 229-245. In: Amphibian Ecology

and Conservation: a Handbook of Techniques. Editor,

Dodd CK. Oxford University Press, Oxford, United

Kingdom. 585 p.

February 2022 | Volume 16 | Number 1 | e304

Adcock et al.

Zachary C. Adcock is a Ph.D. candidate at Texas State University (San Marcos, Texas, USA)

and a Senior Ecologist with Cambrian Environmental (Austin, Texas, USA). Zach received his

B.S. in Biology and Environmental Science from the University of Tampa (Tampa, Florida,

USA) and his MS. in Integrative Biology from the University of South Florida (Tampa, Florida,

USA). His dissertation research addresses the natural history and conservation policy of federally

threatened Jollyville Plateau Salamanders. He is broadly interested in the ecology, conservation, and

management of threatened and endangered wildlife, especially herpetofauna.

Michelle E. Adcock is a Ph.D. candidate at Texas State University (San Marcos, Texas, USA).

Michelle received her B.S. in Environmental Science from the University of Tennessee at Chattanooga

(Chattanooga, Tennessee, USA) and her MLS. in Wildlife Biology from Texas State University. Her

dissertation research focuses on camera trapping techniques, road ecology, and community ecology

dynamics of wildlife in South Texas. She is passionate about wildlife conservation, natural resource

management, and educational outreach.

Bruce E. Hall is an agronomist with Wyman’s (Milbridge, Maine, USA). He received his B.S. in

Biology from the University of Tampa (Tampa, Florida, USA), and serves as Chair of the Wild

Blueberry Advisory Committee and a member of the Penn State University Center for Pollinator

Research Stakeholder Advisory Board. He is generally focused on ecological interdependence and

cross-scale interactions across time and space, and has a wide-ranging passion for community,

economic, and environmental sustainability.

Michael R.J. Forstner is a Professor in Biology at Texas State University (San Marcos, Texas,

USA), and the Alexander-Stone Chair in Genetics. He has a B.Sc. from Southwest Texas State

University, an M.Sc. from Sul Ross State University (Alpine, Texas, USA), and a Ph.D. from Texas

A&M University (College Station, Texas, USA). He has broad interests in the effective conservation

of rare taxa, particularly reptiles and amphibians. The students and colleagues working with him

seek to provide genetic and ecological data relevant to those conservation efforts.

Amphib. Reptile Conserv. 93 February 2022 | Volume 16 | Number 1 | e304

Official journal website:

amphibian-reptile-conservation.org

Amphibian & Reptile Conservation

16(1) [General Section]: 94-105 (e305).

Distribution and habitat suitability of two neighboring

Lycian salamanders

‘Omer Dilbe, 2Akin Kirac, and ‘*Eyup Baskale

‘Pamukkale University, Faculty of Science and Arts, Department of Biology, Denizli, TURKEY ?Canakkale Onsekiz Mart University, Canakkale

Technical Sciences Vocational College, Canakkale, TURKEY

Abstract.—Lyciasalamandra fazilae and Lyciasalamandra flavimembris are two Endangered and endemic

species which occur only in Mugla province of Turkey. In protecting an endemic or endangered species, the

first step is to understand its potential and/or known distribution. Therefore, we used the Maximum Entropy

modelling software (MaxEnt) to analyze the current potential distribution and most important habitat features

associated with the localities of these two species. The variables with the highest contributions to the model

were: Bedrock, Precipitation of Coldest Quarter, and Normalized Difference Vegetation Index for L. flavimembris;

and Bedrock, Temperature Seasonality, Precipitation Seasonality, and Precipitation of Coldest Quarter for L.

fazilae. We also identified two new localities for L. flavimembris using the habitat suitability model.

Keywords. Climate, conservation, Endangered, endemic, habitat suitability map, new localities

Citation: Dilbe O, Kirag A, Baskale E. 2022. Distribution and habitat suitability of two neighboring Lycian salamanders. Amphibian & Reptile

Conservation 16(1) [General Section]: 94-105 (e305).

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.

Accepted: 5 February 2021; Published: 15 March 2022

Introduction on the ecology of Lycian salamanders broadly covers all

Species in this genus, and is therefore considered to be

There are only seven species of Lycian salamanders inthe —— generally applicable to all of them (cf. Ozeti and Yilmaz

world, six of which are found in Turkey. Among them, the 1994). On this basis, the Lyciasalamandra species are

Marmaris Salamander [Lyciasalamandra flavimembris terrestrial, inhabiting rocky limestone areas mostly

(Mutz and Steinfertz 1995)] and the Gécek Salamander in pine forests and maquis—sometimes near single-

[Lyciasalamandra fazilae (Basoglu and Atatiir, 1974)] standing pines and olive trees, sometimes in deciduous

are local endemic species distributed in the Mugla forests dominated by oaks and junipers, and occasionally

province of Turkey. Lyciasalamandra fazilae occurs in accumulations of rocks or on slopes without vegetation

in the eastern part of Mugla province (Fethiye, Gocek, (e.g., Baran and Atatur 1998; Basoglu and Ozeti 1973;

Ortaca, and Koycegiz districts), while L. flavimembris Veith et al. 2001). The vertical distributions of these

occurs in the western part of Mugla province (Milas, species are known to range from 25 to 1,400 m asl,

Ula, and Marmaris districts). Both species were formerly — where the mean annual rainfall may be less than 1,000

considered to be subspecies of Mertensiella luschani, mm (Veith et al. 2001; Yildiz and Akman 2015).

with L. flavimembris even being con-subspecific with Lyciasalamandra fazilae and L. flavimembris are

L. helverseni from the Greek Karpathos archipelago. __ listed as Endangered by the IUCN Red List of Threatened

However, previous studies have shown that they Species (http://www.iucnredlist.org,; Accessed: 4 May

are morphologically and phylogenetically separate 2020) in view of their naturally restricted ranges and

species (Oz et al. 2004; Veith et al. 2016, 2020; Veith the continuing decline of their habitats. In protecting

and Steinfartz 2004), and their colorations are clearly an endemic and/or Endangered species, the first step is

distinguishable (Oz et al. 2004; Ozeti and Yilmaz 1994). to _ understand its potential and/or known distribution

Amphibians are highly susceptible to any changes = (Sousa-Silva et al. 2014). Intense research on the existing

in their habitat because of their highly permeable skin, _ distribution of Lycian salamanders is time-consuming

and many species spend their lives in both terrestrial and = and expensive, but modelling their distributions could

freshwater habitats (Alford and Richards 1999; Barinaga provide more accurate results with less time and effort

1990; Duellman and Trueb 1994). The currentinformation (Hernandez et al. 2006). Species Distribution Modelling

Correspondence. *“eyupbaskale@gmail.com

Amphib. Reptile Conserv. 94 March 2022 | Volume 16 | Number 1 | e305

Dilbe et al.

Pal

> =Bodrum

Marmaris

ie .

er aFethiye

Legend

OM GZS 25 50

Kilometers

e L. fazilae

aL. flavimemris

Fig. 1. Study area and distributions of presence data for L. flavimembris and L. fazilae.

(SDM) is a correlative approach in which habitat

suitability, and therefore the distribution of a species, is

estimated on the basis to environmental and geographical

information (Elith and Graham 2009). The resulting

models are called habitat suitability models, and they

are considered to be important for the conservation of a

species’ habitat and the implementation of conservation

action plans (Buckland and Elston 1993; Marzluff et

al. 2002). They can be used to identify potential risks

to a species and thus to prioritize habitat conservation,

to optimize land management planning, and to allocate

suitable habitats for potential translocation programs

(Corsi et al. 1999; Ozkan and Berger 2014; Stoms et al.

1992).

The effective conservation of amphibian populations

is typically limited by the lack of species-specific

ecological knowledge. Therefore, this study was

conducted to identify the environmental variables which

limit the distribution of Marmaris Salamander and Gocek

Salamander, and to determine their current and potential

habitats. We believe that the models and maps obtained

through the MaxEnt method will provide a base for the

successful execution of species protection action plans.

Materials and Methods

Species data and study area. Between 2012 and 2020,

field studies were carried out during the activity period

Amphib. Reptile Conserv.

of the salamanders (October—April) within the province

of Mugla, Turkey (Fig. 1). The study sites included

four Specially Protected Areas (Gékova SPA, Datca-

Bozburun SPA, Fethiye-Gécek SPA, and K6yceégiz

Dalyan SPA), one National Park (Marmaris NP), and a

Wildlife Development Area (K6ycegiz). The elevations

of the sites ranged from 0 to 1,300 m asl. The climate

is dominated by the Mediterranean climate. Urbanized

areas, touristic areas, and natural areas without human

intervention constitute important places in the study area

which are mostly covered with maquis areas (shrublands),

Red Pine (Pinus brutia) dominated coniferous forest,

and agricultural fields. The field studies were carried out

during both day and night. A total of 240 sample areas

were examined, each with a size of 874 m x 874 m (Le.,

the resolution of the Worldclim [version 2.1] data used

as described below). The altitudes and coordinates of

each salamanders’ presence point were recorded with a

Garmin 62S GPS receiver using the WGS 84 coordinate

system.

Environmental data. The Aster Global Digital Elevation

Model (GDEM), version 3, was obtained from Earthdata

(http://earthdata.nasa.gov). Altitude, aspect, and slope

were produced using GDEM (Zeiler 1999) in ArcMap

10.2 software. The Topographic Position Index (TPI),

Topographic Wetness Index (TWI), Landform Position

Index (LPI), roughness index, hillshade index, ruggedness

March 2022 | Volume 16 | Number 1 | e305

Lyciasalamandra fazilae and L. flavimembris in Turkey

index, solar radiation index, and solar illumination

index (at 0600 h, 0800 h, 1000 h, 1200 h, 1400 h, 1600

h, 1800 h, 2000 h, and total solar illumination) were

created with the help of the “Topography tools” plugin

included in ArcGIS 10.2 (Jenness 2006). The NDVI

(Normalized Difference Vegetation Index) data produced

by the MOD13Q1 module, which is one of the MODIS

VI satellite data sources, was cut and used at the study

area scale. NDVI values range from -1 to +1. Negative

values represent water, snow, clouds, and non-plant

areas; while positive values indicate the presence of

vegetation. However, since negative values complicate

the statistical analysis, the NDVI values were converted

to the 0-10,000 range by using the formula: NDVI *

10,000 (Celik and Gilersoy 2017). The bedrock map of

the study area was obtained from the General Directorate

of Mineral Research and Exploration (Maden Tetkik ve

Arama Genel Mudurluigt, http://yerbilimleri.mta. gov.

tr/anasayfa.aspx). Different bedrock types (154) are

shown in the form of polygons on the digital bedrock

map obtained, which was used as a base map. These

data were used as categorical data. Bioclimatic data

representing the current climatic conditions of the study

area were obtained from http://www. worldclim.org (Fick

and Hijymans 2017). These data (Worldclim, Version 2.1)

were obtained in the WGS 84 coordinate system with

the highest resolution (30 arc-seconds, or 874 m x 874

m), and in the ESRI Grid format. Nineteen bioclimatic

variables (Biol—Biol19, Table 1) with this feature were

cut on the scale of the study area with the help of ArcMap

10.2. Temperature data (Biol, Bio2, Bio5—Bio11) values

are shown multiplied by 100.

For all of the digital base maps of the environmental

variables in ASCII format, each cell was produced in the

WGS 84 coordinate system (874 m x 874 m), and thus is

of the same size as the sample areas.

Statistical evaluation, habitat suitability model, and

habitat suitability model map. Due to the small size of

the study area, high correlation is expected between the

bioclimatic data and other environmental variables. This

may pose a problem during the analysis. To eliminate

the multicollinearity problem, we applied Pearson

Correlation Analysis, using a threshold of r? < 0.8, for a

total of 40 environmental variables. If a pair of variables

was found to have a correlation coefficient greater than

0.8, they were considered to represent related phenomena,

and one of them was excluded from the analysis.

Maximum Entropy (MaxEnt) (Phillips et al. 2006) is

a popular habitat suitability modelling method, which

provides more accurate results with less data in smaller

areas compared with other methods (e.g., DOMAIN,

BIOCLIM, and GARP) (Hernandez et al. 2006; Phillips

and Dudik 2008; Wisz et al. 2008). In addition, MaxEnt

enables the joint processing of categorical and continuous

data (Phillips and Dudik 2008), and it produces a habitat

suitability map (Elith et al. 2011; Hernandez et al. 2006,

Amphib. Reptile Conserv.

2008). MaxEnt is based on ENFA (Ecological Niche

Factor Analysis; Hirzel et al. 2002) and examines the

characteristics of the locations of the target species, and

then estimates a suitability level for all areas based on

the values taken by the factors which affect the known

distribution of the species (Baldwin 2009). In this respect,

the MaxEnt method was used to evaluate the potential

distributions of Marmaris Salamander and Gocek

Salamander using MaxEnt 3.4.1 software (Phillips et al.

2006). MaxEnt calculates the maximum entropy to find

the most likely geographical and ecological distribution of

a target species. MaxEnt also examines the relationships

between the asset data of the target species and

environmental variables, and determines the ecological

requirements of the target species. It then predicts the areas

in which the target species will be more or less likely to

appear based on the ecological requirements of the target

species (Baldwin 2009).

The environmental data, including presence data, in

CSV format and environmental variables in ASCII format

were analyzed with the help of MaxEnt 3.4.1 software.

Species data were separated into 90% for training data

and 10% for test data using the software settings, and

the analysis was adjusted to carry out ten repetitions. The

replicated run type Crossvalidate was selected. Further

settings were: maximum iterations = 500, convergence

threshold = 0.00001, and default prevalence = 0.5.

The Area Under the Receiver Operating Characteristic

(ROC) Curves (AUC) was used to evaluate model

performance. Finally, among the models with excellent

model performance, the model with the lowest standard

deviation between the training data AUC value and the

test data AUC value was selected as “the best model,”

and the species distribution maps of that model were

visualized with ArcMap 10.2 software.

Results

For L. flavimembris and L. fazilae, 83 and 66 presence

data points were obtained from the field studies,

respectively, of which 68 and 54 were used for the final

models, respectively. Most of the presence data obtained

during the field studies were either known localities

or points very close to known localities (Arslan et al.

2018; Baskale et al. 2019; Go¢men et al. 2018; Oguz

et al. 2020; Polat and Baskale 2018; Veith et al. 2020).

According to the results of the habitat suitability model,

the training data set AUC value was 0.942 and the test

data set AUC value was 0.941 + 0.056 (P < 0.001) for

L. flavimembris (Fig 2a); while for L. fazilae the training

data set AUC value was 0.954 and the test data set AUC

value was 0.948 + 0.076 (P < 0.001) (Fig. 2b). These P

values indicated that the model obtained was at the level

of “perfect explanation” for the ecological requirements

in the habitat preferences of both salamanders.

According to the percentages of their contributions to

the MaxEnt model, the important or highly contributing

March 2022 | Volume 16 | Number 1 | e305

Dilbe et al.

yooy AreyUsUTpas

-OWULITOA, “OUOSOWIT'T

-OUO}SpnyA[-sUO}spues

“QUO}SpNyA[-OUO}spues

* oOPLdg “Ouo}SOUN'T

“OHIWIOTO “OU0}SOWI'T

AVoyD “UOrAnTTy

9OUdIINIIO JO [CAIOJUT

€C9

wa)

Som

(%)

[2POP 94} 0} UOTNGI.NUOD

aDv[izVf *T

‘aDpIzbf "T pure SLAQUiaUIAD] "T JO SUOTINGLSIP dy} UTe[Axa Jey} BOUdIINIDO JO S[PAIOJUL DY} YIM JUAXCI] O} SUIPIONIL SO|QLIIVA [EJUSUTUOIIAUD OU} JO SUOTINGINUOD SBVIOAYV *] IIQUL

suelo

“YoYyD ‘UOTANTTY “seta

org ‘gn -yeseg-oupids

“OUIWOTO “oIOpLog

“QUO}SpnyA[-ouUO}spues

-OU0}§ 914qGeq ‘ouUO}SOWT'T

‘uoysoul’T ALIOYD

000°0I

wut OS9—-00S

JOUdLINDIO JO [CAIOJUT

—_—

(%) [SPO 94}

0} UOTINGIyUOD

SLAQUIAUIIADL "T

[eorlosaye_

yeoosayeD

AJOO

AJOD

OO x(LOl/ZOld)

sup

6107 VIN

000C BSEN

9007 Ssouusr

9007 Ssouuor

9007 Ssouusr

9007 ssouusr

9007C Ssouuor

9007C Ssouuer

9007C Ssouuor

6661 21197

6661 91197

6661 121197

L107 SuewT}y pue Yor

L107 SuewlT} pue Yor

L107 Suewt}y pue 391

L107 SuewlT}] pue 391

L107 Suewlt}y pue 391

L107 SueWT] pue 3x1

L107 SuewT}y pue 391

L107 SuewT}] pue 31

L107 Suewlt}y pue 3914

L1OT Suewlt}] pue 391

L107 SuewlT} pue Yor

L107 SuewT}y pue ory

L107 SuewT}] pue 3914

L107 Suewlt}y pue Yor

L107 SuewlT}] pue 3914

L107 SuewT}y pue 391

L107 SuewT}] pue 31

L107 Suewlt}y pue ory

L107 SuewlT}] pue ory

OUDIOFOY

yooIpog

(XOpuU] UONRIOSAA SOUSIOJJIC| POZITEULION) [ACN

(UOTRUTLUNT]I Ie[OS [2}0} pue “Y OOOT “Y COST “Y O09T

‘Y OOF “YU OOTI “YU OOOT “Y 0080 “Y 0090 18) Xepur UOTJeUTLUNITI IejOS

XOPUI UOTCIPeI IeJOS

XOpul ssoupossny

xopul opeys| [TH

XOpul ssouysnoy

(TdT) Xapul uoTIsod wiojspue’T

(TM.L) Xoput ssoujom orydeisodoy,

(Id.L) X9put uoryisod orydessodoy,

podsy

adoj[s

opmynTy

(19LeNC) ISOPTOD JO uoryepAI99I1g) GL OIG

(JoyTeNd sou JO UOTedIoeId) BL Org

(s9yeNC) ISoTIG Jo uOTeUdId91g) / [OI

(91IeNC) ISON9MA JO UOTJeIdIO9Ig) 9 [OIG

(Ajrfeuosvag uoreydioarg) ¢ [org

(qIUOP ISALIG Jo VOHRA) pLOrg

(WIUOP ISONOM JO UOITENIdAIq) €LO1

(uoneydioorg fenuuy) Z1O1g

(1dyeNC) ISOPTOD JO oinyeslodwuiay, usp) [Org

(19yIeNC) SOUL JO DINJeIOdWIDT, URI) OL OT

Joyen() Isoliq jo smmjyeroduiay, ues) 601g

(sayeNC) ISO}OMA JO NyeIOdWIaT, UBATAT) BOTT

(asuey jenuuy oinjeroduiay) Lorg

(WJUO/ ISOP[OD Jo oinjyesodwiay, up) 90g

(QUOT ISOUIVAA JO OINJeIOdWIAT, XPT) SOTg

(Aqyeuoseas oinjyeroduiay) porg

(Aqryeumoyjosy) org

(osuey yeuniq ues) ZOIg

(omnyerodwiay uvsyy enuuy) [org

a1qeiie,

March 2022 | Volume 16 | Number 1 | e305

Amphib. Reptile Conserv.

Lyciasalamandra fazilae and L. flavimembris in Turkey

(a) Sensitivity vs. 1-Specifity for L. flavimembris

Sensitivity (1-Omission Rate)

0.0 0.1 0.2 0.3 0.4

05 0.6

a Training data

a Test data

mw Random Prediction

0.7 0.8 0.9 1.0

1-Specificity (Fractional Predicted Area)

(b) Sensitivity vs. 1-Specifity for L. fazilea

0.5

Sensitivity (1-Omission Rate)

0.0 0.1 0.2 0.3 0.4

= Training data

w Testdata

lw Random Prediction

0.5 0.6 0.7 0.8 0.9 1.0

1-Specificity (Fractional Predicted Area)

Fig. 2. The receiver operating characteristic (ROC) curves for L. flavimembris (a) and L. fazilae (b).

variables which limit the geographical distribution ranges

included three variables for L. flavimembris and four for

L. fazilae (Fig. 3). The percentages of contribution to the

model and occurrence intervals of these environmental

variables are given in Table 1. The variables with the

highest contributions to the model were: Bedrock,

Precipitation of Coldest Quarter, and Normalized

Difference Vegetation Index for L. flavimembris (Table

1 and Fig. 4a); and Bedrock, Precipitation of Coldest

Quarter, Temperature Seasonality, and Precipitation

Amphib. Reptile Conserv.

Seasonality for L. fazilae (Table 1 and Fig. 4b). Combined,

these variables explained 86.3% and 99.1% of the

variation in the two species distributions, respectively.

The habitat suitability models showed the potential

distributions of the two species, and the predicted models

confirmed the mostly known geographical ranges of both

of them (Fig. 5). The area of high predicted probability of

occurrence for L. flavimembris was concentrated around

the Kotekli, Ula, Milas, and Marmaris districts (Fig. 5a).

In particular, the southwestern part of Marmaris district is

March 2022 | Volume 16 | Number 1 | e305

Dilbe et al.

—

Environmental Variable

Bedrock

Bio 15

Bio 19

Bio 4

Environmental Variable

0.80

Jacknife of AUC for L. flavimembris

Without variable |

With only variable ®

With all variables ™

0.90

Without variable ©

With only variable ®

‘| With all variables ®

0.70 0.72 0.74 0.76 0.78 080 082 084 086 088 090 092 0.94 0.96

Fig. 3. Results of the Jackknife test for evaluating the relative importance of environmental variables for L. flavimembris (a) and L.

fazilae (b). See Table 1 for definitions of the environmental variables.

the most intensely occupied area for L. flavimembris. In

relation to the habitat suitability model of L. favimembris,

the field studies revealed two new localities: Kizilkoy

(36°41’°N, 28°06’ E; 204 m asl) in the Selimiye district,

and Icmeler (36°46’N, 28°12’E; 142 m asl) in the

Marmaris district. For L. fazilae, the habitat suitability

model indicated a high probability of occurrence mostly in

known habitats, such as Gokceovacik (Fethiye), Uzimli

(Fethiye), Dalyan (Ortaca), Kapikargin (Dalaman), and

Sultanitye (Koycegiz) (Fig. 5b).

Discussion

According to the MaxEnt results, the average

contributions (in percentage) of the key environmental

variables to the model were determined as: Bedrock

(54.2%), Precipitation of Coldest Quarter (41.7%), and

NDVI (4.1%) for ZL. flavimembris, and Bedrock (62.3%),

Precipitation of Coldest Quarter (25.4%), Temperature

Seasonality (8.5%), and Precipitation Seasonality (3.8%)

for L. fazilae.

The most important factor limiting the distributions

of both L. flavimembris and L. fazilae is bedrock type

rather than any of the climatic conditions. Species that

prefer specific bedrock types need corridors made up of

suitable bedrock to expand their distributions (Sinervo

et al. 2017). It is known that salamanders which live in

suitable bedrock often hide in the cracks, cavities, and

underground of this bedrock under unfavorable climatic

conditions (Baran and Atatur 1998). These cracks

Amphib. Reptile Conserv.

and holes maintain proper moisture and temperature

conditions. The MaxEnt outputs of environmental

variables showed that L. flavimembris prefers 10 of the

154 bedrock types in the region, while L. fazilae prefers

nine bedrock types (Fig. 6). While previous studies

revealed only limestone (Gécmen and Karis 2017; Veith

et al. 2001), this study shows that L. flavimembris and

L. fazilae can be found under different types of stones

but their habitats mostly include limestone and cherty

limestone.

Amphibians have a high climatic sensitivity due

to their ectothermic physiology and their constant

need for moisture (Wells 2007). Previous studies have

emphasized that humid areas, areas with a dense green

cover, an average annual rainfall of 800—1,500 mm, and

rocks with moist ground crevices are suitable habitats

for Lycian salamanders (Baran and Atatur 1998; Veith et

al. 2001). Rodder et al. (2011) investigated the climatic

niche similarities between the Lycian salamander species

using 19 bioclimatic data sets. That study found that

Lycian salamanders (except for L. he/verseni) preferred

similar climatic conditions, and the mean Temperature of

Coldest Quarter (variable Biol1) ranged from 6—12.5 °C

and Precipitation of Coldest Quarter (Bio19) ranged from

350-620 mm. The species-specific studies have shown

that Pinus brutia, Mediterranean maquis, green mosses,

and limestones are indicators for L. flavimembris habitat

(Go¢men and Karis 2017), and the air temperature interval

of the active season of L. flavimembris ranged from 5—21

°C, while monthly average precipitation ranged from

March 2022 | Volume 16 | Number 1 | e305

Lyciasalamandra fazilae and L. flavimembris in Turkey

Coglog Output (Probobility of Presence)

56 64 72 80 89 97 106

Bedrock

1 7 14 22 30 38 46 118 128 139 149

Coglog Output (Probobility of Presence)

8000

10000

NDVI (Normalized Difference Vegetation Index)

-2000 0 2000 4000 6000

—

o =

Coglog Output (Probobility of Presence)

200 =. 250 300 350 400 450 500 550 600 650

Bio19 (Precipitation of Coldest Quarter)

Fig. 4. Variables with the highest contributions to the potential

distributions of L. flavimembris (a) and L. fazilae (b) according

to MaxEnt, with the standard errors in shown blue. In each

graph, the y-axis indicates the probability of presence and the

x-axis shows the contribution of each variable. See Table 1 for

definitions of the environmental variables.

Amphib. Reptile Conserv.

100

(b)

w 6

Oo 5

a 4

a 3

& >

ro)

2 1

56 64 72 80 89 97 107 117 127 137 147

Bedrock

17 #14 22 30 38 46

550 600 650 700 750 800

Bio4 (Temperature Seasonality)

Coglog Output (Probobility of Presence)

So = = YN RP WwW ee Hm

on Oo n Oo nn Oo n Oo on oOo

Bio15 (Precipitation Seasonality)

Coglog Output (Probobility of Presence)

a ne

n Oo n oOo n ao on Oo

f=)

Bio19 (Precipitation of Coldest Quarter)

March 2022 | Volume 16 | Number 1 | e305

Dilbe et al.

Habitat Suitability Map

o Lyciasalamandra flavimembris

Value

— High : 1

10 20

Kilometers

Value

a High : 1

—- Low : 0

0 10 20

Kilometers

Fig. 5. MaxEnt habitat suitability maps for L. flavimembris (a) and L. fazilae (b).

Amphib. Reptile Conserv. 101 March 2022 | Volume 16 | Number 1 | e305

Lyciasalamandra fazilae and L. flavimembris in Turkey

Uk [

& $3 Ss

The number of presence data points

mL. flavimembris

OL. fazilae

re & &

Cc]

S oe S > < in &

ro se Or og Ss iy x we wv

& C SS ye

Bedrock types

Fig. 6. Frequencies of Bedrock types on the different presence points of L. flavimembris and L. fazilae. Bedrock type abbreviations:

Alluvion [All], Breccias [Brec], Pebble Stone-Sandstone-Mudstone [PS-SS-MS], Chert [Cher], Cherty Limestone [Cher LS],

Dolomite [Dol], Limestone [LS], Melange [Mel], Peridotite [Per], Spilite-Basalt-Tuff [SBT], Sandstone-Mudstone [SS-MS],

Sandstone-Mudstone-Limestone [SS-MS-LS], Volcanite-Sedimentary Rock [V-SR], and all unknown rock types [Unknown].

57-335 mm (Baskale et al. 2019). On the other hand,

Polat and Baskale (2018) stated that the greatest number

of individuals of L. fazilae was observed at temperatures

between 2 and 18 °C (mean 12.99 + 0.403 °C), and that

the active period started with the first autumn rains and

a sharp decrease in air temperature (< 20 °C), and ended

with higher air temperatures (22 °C and above).

Climatic conditions may also limit the distributions

of both species, resulting in narrow distribution areas.

Our habitat suitability models show that L. flavimembris

and L. fazilae both have specific demands with respect

to precipitation and temperature. Specifically, the

Precipitation of Coldest Quarter (B1019) is 500-650 mm

for L. flavimembris, while for L. fazilae the Precipitation

of Coldest Quarter (Biol9; 600 mm), Precipitation

Seasonality (Biol5; 100 mm), and Temperature

Seasonality (Bio4; 5—6 °C) were found to be important

predictors of its distribution. These results show that the

current climatic conditions are sufficient for L. fazilae

and L. flavimembris to survive. This supports the MaxEnt

ClogLog values for L. fazilae and L. flavimembris given

in Veith et al. (2020), which showed the prevalence of

unsuitable current climatic conditions for the survival of

many of the Lycian salamanders other than L. fazilae and

L. flavimembris.

In our models, vegetation is another of the

environmental factors that determine the distributions

of the two salamander species. Lyciasalamandra

Amphib. Reptile Conserv.

flavimembris was detected in areas with an NDVI of

10,000, indicating green areas with high canopy cover.

For L. fazilae, the interval of the NDVI value was

wider (1,000—10,000), hence its distribution area is

characterized by more heterogeneous vegetation, such

as pine forests, Mediterranean marquis, and olive tree

fields. Our habitat compatibility model obtained with the

MaxEnt method is compatible with the known biology

of Lycian salamanders (Baran and Atatur 1998; Ozeti

and Yilmaz 1994: Veith et al. 2001). Another consistency

in our results is that the locations with the highest

population densities and abundances of L. flavimembris

and L. fazilae shown in Polat and Baskale (2018) and

Baskale et al. (2019) are the same as the localities with

high suitability values in our habitat suitability map.

Our habitat suitability maps mostly reflect the known

localities of both species, but it is important to consider

some differences between the predicted model and

the known habitats. For L. flavimembris, the habitat

suitability map shows inhabitable areas to the west.

Although the Yalikavak and Mazi Mountain (Milas)

populations (Oguz et al. 2020) are located in this area,

the potential distribution is extended even to the Bodrum

district. This suggests that there are either important

barriers to the species’ dispersion, or it has simply not

yet been recorded from these areas. Moreover, the

model predicted suitable habitats for L. fazilae within

the distribution area of L. flavimembris (see also Veith

102 March 2022 | Volume 16 | Number 1 | e305

Dilbe et al.

et al. 2020). This situation arises from the fact that both

species prefer similar environmental variables such as

Precipitation of Coldest Quarter and Bedrock. On the

other hand, Veith et al. (2020) showed a strong degree of

isolation among Lyciasalamandra populations, including

phyloclades of L. fazilae, and two subspecies of L. fazilae

are recognized: L. f fazilae and L. f ulfetae (Gocmen

et al. 2018). However, Veith et al. (2020) claimed that

the L. fazilae phyloclade diversity is higher than that

reflected by current taxonomy, with five phyloclades

forming three well-supported phylogenetic clusters:

(faz-I + faz-II), faz-III, and (faz-IV + faz-V). The vertical

extension of the Taurus Mountains between the Gocek

and Dalaman districts constitutes the first (faz-I + faz-II)

and the second (faz-III) phylogenetic clusters. However,

the third cluster (Ulemez population and Sultaniye

population) is geographically isolated by the Koycegiz

Lake and Dalyan Canal in the east, and the Ulemez

Mountain and the extensions of Taurus Mountains in the

west and northwest (see Figs. 1 and 5b).

In conclusion, potential distribution maps of L.

flavimembris and L. jfazilae were created based on

bioclimatic data and some environmental variables. These

maps indicated that the current climatic conditions of

the regions where both species live are suitable for the

survival of the species. In addition, some populations of L.

flavimembris (1.e., Yaylasogsut and Aricilar) and L. fazilae

(i.e., Uzimlii and Gokceovacik) were located far from

the Mediterranean coast, indicating that these species can

tolerate more diverse climatic conditions. This study is an

important step for the conservation of endangered species

within and outside existing protected areas, and may help

alleviate the population decline of both species.

Acknowledgements.—This study was a part of the first

author’s M.Sc. thesis. The permission for field work,

handling, and laboratory studies of the salamanders were

issued by the Animal Ethics Committee of Pamukkale

University, Ministry of Forestry and Water Affairs,

General Directorate of Nature Conservation and Natural

Parks, and the Turkish Ministry of Food, Agriculture, and

Livestock. We would like to thank Pamukkale University

Scientific Research Projects Unit-BAP (2013FEBE046

and 2019FEBE062) for their support during this study.

We would also like to thank Hasan Pasali, Ebru Tong,

and Hakan Korbalta for their support in the field studies;

and Ayfer Sirin, Fatih Polat, and Doan Sozbilen for their

assistance in the field and laboratory studies.

Literature Cited

Alford RA, Richards SJ. 1999. Global amphibian

declines: a problem in applied ecology. Annual Review

of Ecology and Systematics 30: 133-165.

Arslan D, Yasar €, Izgin A, Sen C, Cicek K. 2018.

New sites of the Endangered Marmaris Salamander,

Lyciasalamanadra flavimembris (Mutz and Steinfartz,

Amphib. Reptile Conserv.

1995) (Caudata: Salamandridae) from Mu§gla, Turkey.

Amphibian & Reptile Conservation 12: 106-111

(e163).

Baldwin RA. 2009. Use of maximum entropy modeling

in wildlife research. Entropy 11: 854—866.

Baran I, Atatur M. 1998. The Herpetofauna of Turkey

(Amphibians and Reptiles). Republic of Turkey,

Ministry of the Environment, Ankara, Turkey. 214 p.

Barinaga M. 1990. Where have all the froggies gone?

Science 247: 1,033—1,035.

Baskale E, Sozbilen D, Ozyilmaz Y, Dilbe O. 2019.

Population status and threats against Lyciasalamandra

flavimembris (Marmaris Lycian Salamander).

Commagene Journal of Biology 3: 37-43.

Basoglu M, Ozeti N. 1973. Tiirkive Amfibileri. Ege

Universitesi Matbaasi, Izmir, Turkey. 155 p.

Buckland S, Elston D. 1993. Empirical models for the

spatial distribution of wildlife. Journal of Applied

Ecology 30: 478-495.

Celik MA, Gilersoy AE. 2017. Nur Daéglarinin

(Amanoslar) Orta Kesiminin Farkl1 Yamaclarinda

Bitki Indeks Degisiminin Izlenmesi. /Sdir Universitesi

Fen Bilimleri Enstitiisti Dergisi 7: 87-97.

Corsi F, Dupre E, Boitani L. 1999. A large-scale model

of wolf distribution in Italy for conservation planning.

Conservation Biology 13: 150-159.

Duellman WE, Trueb L. 1994. Biology of Amphibians.

Johns Hopkins University Press, Baltimore, Maryland,

USA. 670 p.

Elith J, Graham CH. 2009. Do they? How do they?

Why do they differ? On finding reasons for differing

performances of species distribution models.

Ecography 32: 66-77.

Elith J, Phillips SJ, Hastie T, Dudik M, Chee YE, Yates

CJ. 2011. A statistical explanation of MaxEnt for

ecologists. Diversity and Distributions 17: 43-57.

Fick SE, Hymans RJ. 2017. WorldClim 2: new 1-km

spatial resolution climate surfaces for global land

areas. International Journal of Climatology 37:

4,302-4,315.

Gocmen B, Ehl S, Karis M, Thiesmeier B, Kordges T.

2018. Molecular and morphological evidence for

a new subspecies of Fazila’s Lycian Salamander,

Lyciasalamandra_ fazilae, in South-west Anatolia.

Zoology in the Middle East 64: 304-314.

Gocmen B, Karis M. 2017. Comparative study on

the Endangered Marmaris Lycian Salamander

populations, Lyciasalamandra flavimembris (Mutz

and Steinfartz, 1995) (Caudata: Salamandridae),

with the description of several new localities. North-

Western Journal of Zoology 13: 49-57.

Hernandez P, Franke I, Herzog S, Pacheco V, Paniagua

L, Quintana H, Soto A, Swenson J, Tovar C, Valqui

T. 2008. Predicting species distributions in poorly-

studied landscapes. Biodiversity and Conservation

17: 1,353—1,366.

Hernandez PA, Graham CH, Master LL, Albert DL. 2006.

March 2022 | Volume 16 | Number 1 | e305

Lyciasalamandra fazilae and L. flavimembris in Turkey

The effect of sample size and species characteristics

on the performance of different species distribution

modeling methods. Ecography 29: 773-785.

Hirzel AH, Hausser J, Chessel D, Perrin N. 2002.

Ecological-niche factor analysis: how to compute

habitat-suitability maps without absence data?

Ecology 83: 2,027—2,036.

Jenness J. 2006. Topographic Position Index (tpi_jen.

avx) extension for ArcView 3.x, version 1.3a. Jenness

Enterprises, Flagstaff, Arizona, USA.

Marzluff JM, Millspaugh JJ, Ceder KR, Oliver CD,

Withey J, McCarter JB, Mason C, Comnick J. 2002.

Modeling changes in wildlife habitat and timber

revenues in response to forest management. Forest

Science 48: 191-202.

Oguz MA, Gocmen B, Karis M, Ehl S, Veith M. 2020. Two

new populations of Lyciasalamandra flavimembris

substantially extend the genus’ distribution range in

Anatolia. Biharean Biologist 14: 36-40.

Oz M, Diisen S, Tunc MR, Kumlutas Y, Durmus H,

Kaska Y. 2004. A morphological and taxonomical

study on the subspecies of the Lycian salamander,

Mertensiella luschani (Steindachner, 1891) (Urodela:

Salamandridae). Turkish Journal of Zoology 28: 237-

244.

Ozeti N, Yilmaz I. 1994. Tiirkiye Amfibileri. Ege

Universitesi Fen Fakiltesi Kitaplar Serisi No. 151.

Ege Universitesi, Izmir, Turkey. 221 p.

Ozkan K, Berger U. 2014. Predicting the potential

distribution of plant diversity in the Yukarig6kdere

forest district of the Mediterranean region. Polish

Journal of Ecology 62: 441-454.

Phillips SJ, Anderson RP, Schapire RE. 2006. Maximum

entropy modeling of species geographic distributions.

Ecological Modelling 190: 231-259.

Phillips SJ, Dudik M. 2008. Modeling of species

distributions with MaxEnt: new extensions and a

comprehensive evaluation. Ecography 31: 161-175.

Polat F, Baskale E. 2018. Phenology and factors

influencing the abundance of Lyciasalamandra fazilae

(Amphibia: Salamandridae) in Turkey. Salamandra

54: 123-131.

Rédder D, Lotters S, Oz M, Bogaerts S, Eleftherakos K,

Veith M. 2011. A novel method to calculate climatic

niche similarity among species with restricted

ranges—the case of terrestrial Lycian salamanders.

Organisms Diversity and Evolution 11: 409-423.

Sinervo B, Lara Reséndiz RA, Miles DB, Lovich JE,

Amphib. Reptile Conserv.

Ennen JR, Miller J, Cooper RD, Rosen PC, Stewart

JA, Santos JC, et al. 2017. Climate Change and

Collapsing Thermal Niches of Mexican Endemic

Reptiles. University of California Press, Berkeley,

California, USA. 21 p.

Sousa-Silva R, Alves P, Honrado J, Lomba A. 2014.

Improving the assessment and reporting on rare and

endangered species through species distribution

models. Global Ecology and Conservation 2: 226—

237.

Stoms DM, Davis FW, Cogan CB. 1992. Sensitivity of

wildlife habitat models to uncertainties in GIS data.

Photogrammetric Engineering and Remote Sensing

58: 843-850.

Veith M, Baran I, Godmann O, Kiefer A, Oz M, Tung

MR. 2001. A revision of population designation and

geographic distribution of the Lycian salamander

Mertensiella luschani (Steindachner, 1891). Zoology

in the Middle East 22: 67-82.

Veith M, Go¢men B, Sotiropoulos K, Eleftherakos K,

Lotters S, Godmann O, Karis M, OSuz A, Ehl S. 2020.

Phylogeographic analyses point to long-term survival

on the spot in micro-endemic Lycian salamanders.

PLoS ONE 15: e0226326.

Veith M, Gocmen B, Sotiropoulos K, Kieren S, Godmann

O, Steinfartz S. 2016. Seven at one blow: the origin of

major lineages of the viviparous Lycian salamanders

(Lyciasalamandra Veith and Steinfartz, 2004) was

triggered by a single paleo-historic event. Amphibia-

Reptilia 37: 373-387.

Veith M, Steinfartz S. 2004. When non-monophyly

results in taxonomic consequences: the case of

Mertensiella within the Salamandridae (Amphibia:

Urodela). Salamandra 40: 67-80.

Wells K. 2007. The Ecology and Behavior of Amphibians.

University of Chicago Press, Chicago, Illinois, USA.

1,400 p.

Wisz MS, Hiymans R, Li J, Peterson AT, Graham C,

Guisan A, Group NPSDW. 2008. Effects of sample

size on the performance of species distribution

models. Diversity and Distributions 14: 763-773.

Yildiz MZ, Akman B. 2015. A new subspecies of Atif’s

Lycian Salamander, Lyciasalamandra atifi (Basoglu,

1967), from Alanya (Antalya, Turkey). Herpetozoa

28: 3-13.

Zeiler M. 1999. Modeling Our World: the ESRI Guide

to Geodatabase Design. ESRI, Redlands, California,

USA. 297 p.

March 2022 | Volume 16 | Number 1 | e305

Amphib. Reptile Conserv.

Dilbe et al.

Omer Dilbe has graduated from the M.Sc. program of the Department of Biology at the

University of Pamukkale (Denizli, Turkey). He is currently a researcher in species/ecosystem

ecology and conservation.

Akin Kirag¢ is an Associate Professor in the Technical Science Vocational School at Canakkale

Onsekiz Mart University, Canakkale, Turkey. His primary research interests mainly focus on

habitat suitability models and climate envelope models, and he has conducted research in the field

of species ecology and climate change. He has authored or co-authored several articles in peer-

reviewed scientific journals on insects, amphibians, reptiles, and mammals.

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

a special focus on the conservation of amphibians and reptiles living in Turkey. Eyup currently

coordinates several research projects focusing on threatened species in Turkey.

105 March 2022 | Volume 16 | Number 1 | e305

Amphibian & Reptile Conservation

16(1) [General Section]: 106-135 (e306).

Official journal website:

amphibian-reptile-conservation.org

Turtles of Colombia: an annotated analysis of their diversity,

distribution, and conservation status

1*Vivian P. Paez, ‘Brian C. Bock, ‘Diego A. Alzate-Estrada, **Karla G. Barrientos-Munoz,

‘Viviana M. Cartagena-Otalvaro, *°Andrea Echeverry-Alcendra, ‘Marley T. GOmez-Rincon,

6Cristian Ramirez-Gallego, ‘Jennifer Sofia del Rio, and *Margarita M. Vallejo-Betancur

'Grupo Herpetologico de Antioquia, Instituto de Biologia, Facultad de Ciencias Exactas y Naturales, Universidad de Antioquia, Calle 67 # 53-

108, Bloque 7-121, A.A. 1226, Medellin, COLOMBIA *Fundacion Tortugas del Mar, Calle 46 A sur # 40 A-31 Interior 103, Envigado, Antioquia,

COLOMBIA ?Wider Caribbean Sea Turtle Conservation Network - WIDECAST, Calle 46 A sur # 40 A 31 Interior 103, Envigado, Antioquia,

COLOMBIA *Departamento de Biologia y Conservacion, Fundacion Botanica y Zoologica de Barranquilla-FUNDAZOO, Calle 77 # 68 — 40,

Barranquilla, Atlantico, COLOMBIA *Asociacién Colombiana de Parques Zooldgicos, Acuarios y Afines-ACOPAZOA, Barranquilla, Atlantico,

COLOMBIA °Corporacion para el Desarrollo de la Costa Caribe-CORPOCARIBE, Cartagena, Bolivar, COLOMBIA 'Grupo de Ecologia Evolutiva

y Biogeografia Tropical-ECOBIT, Universidad INCCA de Colombia, Carrera 13 No. 24-15, Bogota, D.C., COLOMBIA *Fundacion Ecolombia,

Calle 5° # 35-87 Interior 1102, Medellin, Antioquia, COLOMBIA

Abstract.—With this analysis, we update the state of knowledge on the species richness, distribution, and

conservation status of the turtles of Colombia, both at the national level and regionally within Colombia by

hydrological drainages and geopolitical distribution units (departments). The richness patterns and conservation

status are analyzed at taxonomic and geographic levels, and the implications of the description of new species on

our knowledge of their distribution and conservation status in the country are discussed. Finally, annotations are

given on the turtle species that have been introduced into Colombia, translocated within the country, erroneously

reported, or deemed to be taxonomically invalid. Our conservative analysis in terms of richness (based upon

validated occurrence records) confirms that there are 33 species and two subspecies of turtles in Colombia, of

which five are sea turtles and 28 are tortoises or freshwater turtle species. Colombia has 17 genera of chelonians

in nine families, so it is second behind Brazil in terms of the number of extant species in South America. The

proportion of threatened species in Colombia exceeds 43%, and the threatened species are not evenly distributed

among higher taxa or regions. Commonalities were found in the national conservation status assessments for

most of the turtle species shared among the five most species-rich countries in South America, including sea

turtles and podocnemidids (except for the podocnemidids in Brazil).

Keywords. Chelonians, endemism, Reptilia, sea turtles, South America, threats, tortoises

Resumen.—En este analisis actualizamos el estado de conocimiento sobre la riqueza de especies, distribucion y

estado de conservacion de las tortugas de Colombia, tanto a nivel nacional, como por cuencas hidrologicas y por

distribucion geopolitica (departamentos). Analizamos los patrones de riqueza y conservacion a nivel taxonomico

y geografico, y discutimos las implicaciones de la descripcion de especies nuevas en el conocimiento de

su distribucion y conservacion en el pais. Finalmente, hacemos anotaciones sobre las especies de tortugas

introducidas, traslocadas a nivel nacional, erroneamente reportadas, o consideradas taxonomicamente

invalidas. Nuestro analisis conservador a nivel de riqueza de especies (basados en registros de ocurrencia

validados) confirma que en Colombia ocurren 33 especies y dos subespecies de tortuga, de las cuales cinco

son marinas y 28 son terrestres o de agua dulce. Colombia cuenta con nueve familias de quelonios, 17 géneros

y es segundo en Suramerica después de Brasil en terminos del numero de especies vivientes. La proporcion de

especies amenazadas en Colombia excede el 43% y no se distribuye equitativamente por familia o por regiones.

Encontramos similitudes en las evaluaciones nacionales de los estados de conservacion para la mayoria de las

especies de tortugas compartidas entre los cinco paises de mayor riqueza del orden en Suramerica, incluyendo

a las tortugas marinas y los podocnemididos (con la excepcion de los podocnemididos de Brasil).

Palabras clave. Amenaza, distribucion, diversidad, endemismos, quelonios, Reptilia, Suramérica

Citation: Paez VP, Bock BC, Alzate-Estrada DA, Barrientos-Mufoz KG, Cartagena-Otalvaro VM, Echeverry-Alcendra A, G6mez-Rincon MT, Ramirez-

Gallego C, del Rio JS, and Vallejo-Betancur MM. 2022. Turtles of Colombia: an annotated analysis of their diversity, distribution, and conservation status.

Amphibian & Reptile Conservation 16(1) [General Section]: 106—135 (e306).

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.

Accepted: 20 September 2021; Published: 7 April 2022

Correspondence. *vivianpaez!@gmail.com

Amphib. Reptile Conserv. 106 April 2022 | Volume 16 | Number 1 | e306

Paez et al.

Introduction

The geographic range of a species is determined by

a combination of ecological and historical factors,

irrespective of political borders (Gaston 2003). However,

species inventories are usually conducted for areas

defined by artificial boundaries to produce species lists

for specific protected areas or political regions. For

example, national checklists have been used to identify

“mega-diverse” countries, which can help donor agencies

and conservation organizations prioritize their efforts to

preserve biodiversity. Updates of national checklists may

also help to document increases in the known species

richness of a country, thereby providing a metric on the

rate of growth in knowledge for a particular taxonomic

group. National checklists provide an important first step

in identifying species that face conservation concerns,

because the responsibility for enacting and enforcing

conservation legislation and actions lies with institutions

at the national level.

Colombia is one of the richest countries in terms of

turtle species, and thus plays an important global role in

their study and conservation. Ceballos-Fonseca (2000)

published the first checklist of turtles for Colombia,

which has been followed by other updates in the literature

during the past two decades (Paez et al. 2012a; Morales-

Betancourt et al. 2015a; Forero-Medina et al. 2016). The

richness of the Colombian turtle fauna has also been

summarized in several global turtle species compilations,

such as Uetz et al. (2021) and the nine editions of the

Turtle Taxonomy Working Group checklists (TTWG

2021). Checklists should be updated frequently,

especially for countries like Colombia where the effort

of biodiversity studies, including turtles (Bock and Paez

2017), has increased exponentially in recent years.

The different checklists have all shown that Colombia

and Mexico are the two countries with the most families

of turtles (nine), and they rank among the top one-

fifth globally in terms of turtle species richness (most

recently, 33 species; TTWG 2021). However, these

checklists have either failed to detail the sources of the

records they were based upon or admitted to having

used databases comprised of multiple kinds of records.

However, voucher types vary considerably in terms of

scientific merit (Lehn et al. 2007), from anecdotal reports

of sightings found in gray literature reports, to e-vouchers

such as photographs available on the Internet, to those in

the published peer-reviewed literature, and catalogued

museum voucher specimens.

For this reason, the purpose of this study was to provide

an updated checklist of the turtle species of Colombia,

including information on their distribution within the

country, based upon only the most scientifically solid

evidence available. Thus, the information in this catalogue

is limited almost exclusively to data from catalogued

museum voucher specimens or from peer-reviewed

publications in the scientific literature. Annotations are

Amphib. Reptile Conserv.

provided in cases where taxonomic issues exist, as well as

comments on the conservation status of new or otherwise

non-evaluated taxa. Finally, analyses of the occurrence

data are presented which compare the turtles found in

different hydrological drainages and geopolitical units

(departments) within Colombia, as well as comparisons

of the species richness and conservation status of turtles

in Colombia with the other turtle-rich countries in South

America.

Materials and Methods

Previous checklists of Colombian turtles have sum-

marized species distributions within the country either by

hydrographic drainage (because most Colombian turtles

are freshwater species) or by geopolitical distribution

units (departments). This catalogue documents the

occurrence of Colombian turtle species by department

and by hydrographic drainage, with the latter based on

the five macro-drainages recognized by the Instituto

de Hidrologia, Metereologia y Estudios Ambientales

(IDEAM 2013; Fig. 1): Amazon (AMA), Orinoco (OR),

Caribbean (CAR), Magdalena-Cauca (MAG-CAU), and

Pacific (PA). IDEAM considers the MAG-CAU drainage

as distinct from the other Colombian rivers that drain into

the Caribbean (CAR) due to its much greater discharge

rate. Sea turtle species were excluded from the analyses

involving these freshwater macro-drainages, but not

from the analyses involving departments.

First, the most recent nomenclatural listings of turtle

species proposed by the TTWG (2021; and previous

editions) were examined, along with the slightly different

taxonomic scheme used by the online Reptile Database

(Uetz et al. 2021). For all taxa that have been purported

to occur in Colombia, voucher specimens were identified

in various biological collection databases by accessing

the websites of the Global Biodiversity Information

Facility (GBIF.org 2021), HerpNET (HerpNET 2021),

and the Sistema de Informacion sobre Biodiversidad de

Colombia (SiB Colombia 2021). We attempted to locate

at least one voucher specimen for each macro-drainage,

as well as for each of the departments in Colombia where

a species had previously been reported to occur. When

multiple voucher specimens for the same department

were located, the oldest voucher specimen was chosen

for citation here as evidence of occurrence. This was

done to minimize the risk of including specimens that

were translocated to a location outside of their natural

range, given the frequent releases of confiscated turtles

by Colombian authorities in recent years, often with no

knowledge of the provenance of the individuals being

released (Morales-Betancourt et al. 2012a).

Vouchers of the Colombian turtle species were

found in the following collections, using the museum

acronyms of Leviton et al. (1985) and Iverson (1992):

AMNH: Herpetology-R (American Museum of Natural

History Herpetology Collections), ARAUQ (Coleccion

April 2022 | Volume 16 | Number 1 | e306

Turtles of Colombia: diversity, distribution, and conservation

75°0'0"O

ECUADOR

PERU

ees Kilometers

0 50100 200 300 400

Coordinate system: UTM 18N

70°0'0"O

VENEZUELA

Hydrographic drainages

c=) Amazonas

[| Caribe

= Magdalena-Cauca

Orinoco

C Pacifico

75°0'0"O

70°0'0"O

Fig. 1. Map indicating the locations of the five macro-drainages used in this study IDEAM 2013).

de Anfibios y Reptiles de la Universidad del Quindio),

CBUMAG:REP (Coleccion Biologica de la Universidad

del Magdalena: Reptiles), COLZOOCH-H (Coleccion

Cientifica de Referencia Zoologica del Choco-

Herpetologia), CVS (Reptiles Corporacion Autonoma

Regional de los Valles del Sint y del San Jorge), FMNH

(Field Museum of Natural History (Zoology) Amphibian

and Reptile Collection), HERPETOS-UQ (Coleccion de

Herpetologia de la Universidad de Quindio), [AVH-R

(Coleccion de Reptiles del Instituto de Investigacion de

Recursos Biologicos Alexander von Humboldt), [AVH-

CT (Coleccion de Tejidos del Instituto de Investigacion

de Recursos Biologicos Alexander von Humboldt),

ICN-MHN-Rep o ICN (Coleccion de Herpetologia

del Instituto de Ciencias Naturales de la Universidad

Nacional de Colombia), KUH (University of Kansas

Amphib. Reptile Conserv.

Biodiversity Institute Herpetological Collection),

MCNUP-H (Coleccion Herpetologica del Museo de

Ciencias Naturales de la Universidad de Pamplona),

MHNU-H (Coleccion Herpetologica-Museo de Historia

Natural Unillanos); MHUA-R (Coleccién de Reptiles,

Museo de Herpetologia de la Universidad de Antioquia),

MLS-quel (Coleccién de Quelonios Museo de La Salle

Bogota), UIS-MHN (Coleccion Herpetologica del

Museo de Historia Natural de la Universidad Industrial

de Santander), MPUJ-REPT (Coleccion de Reptiles del

Museo de Historia Natural de la Pontificia Universidad

Javeriana), MVZ-Herp (Museum of Vertebrate

Zoology, University of California Berkeley), SINCHI-R

(Instituto Amazonico de Investigaciones Cientificas

SINCHI), SMF (Forschungsinstitut und Natur-Museum

Senckenberg), USNM (National Museum of Natural

April 2022 | Volume 16 | Number 1 | e306

Paez et al.

History, Smithsonian Institution), and UV-C (Coleccion

de Anfibios y Reptiles de la Universidad del Valle).

To fill the gaps for macro-drainages or departments

where no voucher specimens were found, the peer-

reviewed scientific literature on Colombian turtle species

was consulted. The book Biologia y Conservacion de las

Tortugas Continentales de Colombia (Paez et al. 2012a) is

a comprehensive edited volume with over 40 contributing

authors that summarized the state of knowledge, at that

time, on the tortoise and freshwater turtle species in

Colombia. An analysis of the bibliography included

in that book (Bock and Paez 2017) found that 269

citations involved studies conducted within Colombia

on one or more of its native turtle species. We used this

database and complimented it with our personal working

bibliographies of publications on sea turtles in Colombia,

as well as the more recent publications on any turtle

species in Colombia, to yield a comprehensive source

of solid evidence for the occurrence of Colombian turtle

species in different macro-drainages and departments.

We also incorporated our own personal records from

working on these species in Colombia. Finally, as a

secondary means to corroborate the distribution of

Colombian turtle species based upon literature records,

for departments where our only evidence of presence

was a published article, we also consulted iNaturalist

(including non-research grade records; iNaturalist 2021)

for photographs of those species that show taxonomically

useful characters and were accompanied by geographic

coordinates.

For the data analysis, the comparisons of the turtle

communities occurring within the five macro-drainages

were conducted with a cluster analysis of the occurrence

data, using a grouping analysis based on Jaccard indices

with the vegan package (Oksanen et al. 2007) in R (R

Core Team 2016). To summarize species richness data by

department, maps were generated based on cartographic

information obtained from the databases of the Instituto

Geografico Agustin Codazzi (IGAC 2021) and the

Sistema de Informacién Ambiental de Colombia (SIAC

2021). The data were standardized in terms of format,

coordinate system, scale, and resolution, and processed

with ArcGIS software (version 10.4) (ESRI 2014).

Results

Species richness and distribution

The presence of 35 taxa (33 species, two of which

included two subspecies each) in Colombia was

documented based on both voucher specimens and the

published literature (Table 1). The lone exception was

our failure to locate vouchers or literature records for

the recently resurrected species Mesoclemmys wermuthi

(but see the Recent taxonomic changes section below).

If one accepts that 14 wermuthi is present in Colombia,

then Colombia contains populations of a total of five sea

Amphib. Reptile Conserv.

turtle species, 28 tortoise and freshwater turtle species,

and one exotic species (Trachemys scripta elegans). The

native Colombian species belong to both suborders (16

Pleurodira species and 17 Cryptodira species) divided

among nine families and 17 genera. Colombia has four

endemic turtle species: Kinosternon dunni, Mesoclemmys

dahli, Podocnemis lewyana, and Trachemys medemi. The

most speciose families are Chelidae with nine species

and Podocnemididae with seven species, constituting

48.5% of the Colombian turtle fauna.

In terms of the distributions of these species within

Colombia, both vouchers and scientific publications

confirmed the previously reported occurrences of turtle

species in the five macro-drainages. However, at the

level of departments, only the scientific publications

confirmed all previously reported occurrences. Among

the total of 263 occurrence reports for Colombian turtle

Species in specific departments, voucher specimens could

not be located to corroborate the evidence from scientific

publications in 38% of the cases. Among those cases, 14

reliable observations were found in iNaturalist to help

corroborate the scientific literature reporting the presence

of a species in a department.

Among the macro-drainages, CAR possesses the

highest species richness (14 species), followed by OR

and AMA (13), MAG-CA (eight), and PA (six) (Fig.

1). Nine species (Chelus fimbriata, Chelus orinocensis,

Mesoclemmys wermuthi, Podocnemis erythrocephala,

Podocnemis sextuberculata, Podocnemis vogli, Rhino-

clemmys diademata, and Trachemys medemi) only occur

in one of the five macro-drainages. In terms of species

compositions, the two macro-drainages located east

of the Andes (OR and AMA) differed from the three

Andean macro-drainages (CAR, PA, and MAG-CA),

with a dissimilarity of 95% (Fig. 2). The most similar

macro-drainages were OR and AMA (56%), with nine

shared species, and CAR and MAG-CA (60%), with eight

shared species (Table 1). The MAG-CA macro-drainage

lacked any unique species. The PA macro-drainage was

grouped with the CAR and MAG-CA cluster, but with a

low similarity (35%), and it shared seven species with

CA and four of its species also occupy the MAG-CA

macro-drainage.

Eight departments make up Colombia’s Caribbean

coastline, plus the island department of San Andres,

Providencia, Santa Catalina, while the Pacific coastline of

Colombia is divided among four departments. While all

five of Colombia’s sea turtle species have been documented

to forage and nest in some of Colombia’s departments,

two species are restricted to only one coastline: Caretta

caretta in the Caribbean and Lepidochelys olivacea in the

Pacific. The Magdalena and Choco departments present

the highest documented nesting species richness for sea

turtles (four species each).

Among the non-marine turtles of Colombia,

our analysis found that four species (Chelonoidis

carbonarius, Kinosternon leucostomum, Kinosternon

April 2022 | Volume 16 | Number 1 | e306

Turtles of Colombia: diversity, distribution, and conservation

MAG-CAU

CAR

AMA

1.0 0.8 0.6

0.4 0.2 0.0

Dissimilarity

Fig. 2. Cluster diagram comparing turtle community species compositions of the five macro-drainages in Colombia.

scorpioides, and Rhinoclemmys melanosterna) have

the widest distributions, occupying from 17 to 20 of

Colombia’s departments; while at the other extreme are

seven species with restricted ranges within the country:

Mesoclemmys wermuthi and Rhinoclemmys diademata

only occur in one department, Kinosternon dunni and

Trachemys medemi only occur in two departments, and

Chelus orinocensis, Podocnemis erythrocephala, and

Rhinemys rufipes only occur in three departments. The

departments with the highest species richness (including

marine turtles) are Amazonas with 13 species, and

Antioquia, Caqueta, Chocd, Cordoba, Guainia, Meta,

and Vichada with 12 species each. At the other extreme,

Huila, Norte de Santander, Quindio, Risaralda, and

Tolima each have only three species or less (Fig. 3).

Of the 33 Colombian turtle species, only four

(Chelus fimbriata, Chelus orinocensis, Rhinoclemmys

diademata, and Rhinoclemmys nasuta) have museum

voucher specimens which document their occurrence in

all departments within their ranges. The species with

the most poorly documented distributions in Colombia

include one of the endemic species, Mesoclemmys dahli

(only two of the six departments where it occurs have

vouchers), Peltocephalus dumerilianus (only two of the

nine departments), Podocnemis sextuberculata (only

one of four departments), and all five sea turtle species

(Chelonia mydas, two of 11 departments; Eretmochelys

imbricata, four of 11; Dermochelys coriacea, two

of 10; Caretta caretta, two of five; and Lepidochelys

olivacea, two of four). We do not know if the only

voucher of Mesoclemmys raniceps (from only one of

the five departments where it occurs) truly belongs to

this species or 1s actually a specimen of M. wermuthi.

Amphib. Reptile Conserv.

Erroneous reports and species otherwise excluded

from this checklist

Chelonia agasizzi (Bocourt, 1868). Ceballos-Fonseca

(2000) listed Chelonia agasizii as having a distribution

that includes the Pacific coast of Colombia. However,

the consensus since then has been that there is no

justification for recognizing the “black sea turtle” as a

valid species, but rather that these populations simply

constitute somewhat distinctive populations of the

Green Sea Turtle, C. mydas (Karl and Bowen 1999;

TTWG 2017).

Chelonia mydas (Linnaeus, 1758). Ceballos-Fonseca

(2000) stated that this species nests on both the

Caribbean and Pacific coasts of the Choco Department,

but in the Caribbean portion of the Chocé Department

no nesting by this species has been documented, with

sightings limited to juvenile individuals foraging in

marine grasses along the coast (C. Ramirez-Gallego and

K.G. Barrientos-Mufioz, pers. comm.). Nesting by this

species on the Pacific coast of the Chocé Department is

sporadic (Barrientos-Mufioz et al. 2013).

Eretmochelys imbricata (Linnaeus, 1766). Ceballos-

Fonseca (2000) reported that this species nests on

both coasts of Colombia. However, nests have only

been documented on Caribbean beaches in Colombia,

while nesting by this species in the Pacific has not been

recorded, although there have been a few sightings

of individuals foraging there (Barrientos-Mufioz et

al. 2015a, 2020; Gaos et al. 2010; Tobon-Lopez and

Amorocho 2014; Trujillo-Arias et al. 2014).

April 2022 | Volume 16 | Number 1 | e306

Paez et al.

75°0'0"O

10°0'0"N

5°0'0"N

0°0'0"

70°0'0"O

10°0'O"N

VENEZUELA

5°0'0"N

BRASIL

0°0'0"

NUMBER OF SPECIES

ECUADOR ome

|_£

: ma’

PERU —F

ees Kilometers ma

0 50100 200 300 £400 |

Coordinate system: UTM 18N |

5°0'0"S

5°0'0"S-&

75°0'0"O

70°0'0"O

Fig. 3. Map indicating turtle species richness by department in Colombia.

Lepidochelys kempii (Garman, 1880). Ceballos-

Fonseca (2000) listed Lepidochelys kempii as occurring

along the Caribbean coast of Colombia. However, the

range of this species has been characterized as limited

to coastal habitats of the northern Gulf of Mexico and

northwestern Atlantic Ocean (Mexico and the USA;

Marquez 1990; TTWG 2017), with occasional sightings

of individuals in the northeastern Atlantic (Bolton

and Martins 1990; Covelo et al. 2016) and even in the

Amphib. Reptile Conserv.

Mediterranean (Insacco and Spadola 2010). While it is

possible that Lepidochelys kempii individuals occasional

wander into Colombian waters, we found no voucher

Specimens or reports in the peer-reviewed literature to

support this possibility, so this species was not included

in the current checklist.

Lepidochelys olivacea (Eschscholtz, 1829). Ceballos-

Fonseca (2000) reported that this species occurs and nests

April 2022 | Volume 16 | Number 1 | e306

distribution, and conservation

Turtles of Colombia: diversity,

SEs doy

-NHI-NOI ‘ONS ‘9799 dey-NHW-NOI ‘NVS

‘TOOLT U-VNHW ‘OVW ‘70000 dau ‘OVNINEO

‘OVI ‘S92 dey-NHW-NOI “MOO “9Sz fanb

-STW ‘SAD ‘900L1 U-VNHW ‘TOR “SSL

d3y-NHW-NOI “TLV ‘vIE8l Y-VAHW °LNV

8E9861

WNSN ‘OWA ‘0SZL 4ea-NHW-NOI “AWN ‘SZSL

doy-NHW-NOI ‘OHO “26 fenb-s TW :AVO

SZ [enb-S TW ‘OVA ‘871 £699¢/SUOTeAIASqO/S10

‘ystTeInyeur erquiojoo//:sdyy :ONS ‘6S0r NHN

-SIN (NVS ‘0€7 ONUV ‘ON-SOLAdYAH INO

“POLL d9Y-NHW-NOI “AWN ‘7999 dox-NHW

-NOI ‘OVW ‘8€1 SATILAAY ‘SAO “WOO “8891

doy-NHIW-NOI ‘OHO ‘OL0TTTS9/suonearasqo

/310 71sI]eINyeul eIquIO[Os//:sdyY :SAO ‘89E

O-NAL ‘VO $098876/SUOTeAIOSq0/IO

‘ysI]eINyeur eIquIojoo//:sdyy -AOW ‘1891 doy

-NHW-NOI -TOd -€cI8 1 Y-VAHW -LNV

681 fenb-S TW -VSN

THT [OND-S TW ‘OVA “6I1SL dou

-NHICNOI ‘UVN “S9TT H-HOOOZTOO ‘OHO

“€9960I-U NHINV ‘NVO ‘€€T7ZI U-VNHW :LNV

11z fanb

-STW ‘OVA ‘81799 dou-NHI-NOI ‘INO “SEEz

doy-NHW-NOI ‘OHO ‘€10L1 U-VNHW :LNV

S1aYINOA

LIOT Te 10 ZOMWIRY-SBILA “O[QZ ‘[B 10 OID

-eyedeZ ‘9107 SoI0D-euoIeg pure oj[Mueler

-Z910] J SS TOT O[PASTV-SeUudpIeD pue jasuey

-RUIPIW “PLOT [e 19 vaOD-SaUOW “VIOT

‘ye 10 odansay ‘ETOT Te 19 SoMO]L-oourlg

“S1OT “BTIOT Te 19 YOO “1107 josuey

-BUIPIL “GZO0T Wapayl puv BIOP[-OURISeD

6107 “ELOT [2 19 odansoy

“SPIED “QT1OT ‘1B 19 OPIRIID “Z10T [9 UeD

“6007 PLISOTV pur ZO19g ‘8OOT SBUI[ES-sesIeA,

PUB LIDIIOH]-ONSED “POOT ‘[e 19 BIOP-OURISeD

0c0c Te

J9 OZBIF-OJUIY “610T OURPUSAV-zoeZUOD

“6107 O[]080.-[eleareD ‘9107 SpLIOQ-euOIeg

pure ofits f-ZoI0],J SE [OT ‘Te 19 ZoMWIeY

-SUBIVA “ETIOT Je 19 BIOIVH-ALIOADYIT 1 [0T

JesueY-LUIPS|[ “8OOT SCUT[ES-sesIeA pure

PIOIIOH]-ONSCD “POOT [eB 19 BIO-OURISeD

QS 107 “BZ LOT OSse’'T

pue yNooULJOg-So[LIOW {[ [OT ‘[e 12 OJSOULIV

BE107

‘PTIOT TE 12 OPI “gO0T SBUTTS-sedIEA,

pur eIOIIOH-ONSeD “POT ‘Te 19 BIOP[-OURISeD

L10¢ |e

OULZO'T-O8URIY *TI OT ‘Je 10 Soleqe-opeesoy

“TOOT Te 19 Z9¥d “GZOOT BIOW-OURISeD

“966| BuUlled-ziny pue oeusx °//6] Wapey|

UOHNQALYSIP IYdvV1G0IS 10} S9IUIAIJOY

OVA

IN 007 “UVN ‘OHO ‘NVO

snjv}s

UONvAAISUOD

[euoneN

awepuq

ONS ‘NWS

‘OVI “OVT ‘NNO

“YOO “SAO “TWO

“TO “TLV “LNV

OVA ‘ONS

‘NVS ‘INO “UVN

‘OVI ‘OVT ‘NAO

YOO “OHO “SAO

‘NVO “TVO ‘AOd

“TOE “TLV “LNV

OVA UVN WOO

“OHO ‘NVO ‘LNV

YIVN ‘WOO ‘OHO

‘NVO “TIVO “LNV

wornqgiysip

[ear}odoa5

sosvulevip 1ydeiso1pAy Ul UOINGLysIg

BOLIOUTY

‘LS 7 LP6l

HNWG

“Ogg AeID

SIST onbsouyey

Jopenoq

“sepyerousy,

“LS-PS SE LV6I

HNWA ‘7061

Josuajnog

eIqUIO[OZ

‘ude Jap OJJOH

SPE Lrél

HNWG

198 Abin

ejONZoUd,,

‘oqreoelep|

“I18r AINS

PS6] SUSHOI

‘

Jopenoy

“sepyerous7,

‘9S TT 1 9P6l

HNN

‘0981 ABD

8981 PleqoouL

Jopenoq

“Tnbedeny

‘00SSr ANZ

‘TIBI SIOed

Ayrpesoy ad 4}

pue ‘adAjojoy

“loyny

‘poydaodqnjd sduajv] JO Satdadsqns OM} SY} JO JOYII9 0} SUOTIG ABU JOYONOA « »

(osedeyen

“RdJOOIH{ “IOPI[S UkIqUIO[OD)

SLASOAMJI]DI DISNUAA SAUAYIDAL “9

oeprpAurg ApUre]

(euROdOYD ON

op eSNOL ‘apin[, poo posou

-O8IU']) DINSDU SduUAaIOULYY *¢

(OWIO “eIOWed

‘OTN, POoM ueIqQuIO[OD)

DUAAJSOUDJAU SAUUAJIOUIYY “py

(esuonsuy ‘OIsaN OSede[eH

SOTUNL, POOMA OGreovieyy)

pyowappip sXuualzoulyYy

(e1oquieg

“eIoURIUO|Y ‘OTN, poo UMOIg

pDjnuuD SdwuUajIOUulyY “TZ

aepipAwio0ssy ApIWe yy

(ayoeg ‘ourduig

‘opin Suiddeus uesiioury

YInos) snysouynon DApA[ayD ‘|

oeplupAjayD Aqrmey

BaIpoydAay 1aps0qns

(vIquIO,O, UT pasn sue pue

Ysi[suy Ul sWeU UOWIWIOD) UOXxe],

‘TYINUAIM "VY

JO sdaaiupd PW 01 puodsos109 ABU JOYONOA x *SAIYINOA “poyenyeaq JON “AN pue “UIIDUO|D ysKd'T “D’T “WUaIDYaq eed “dq ‘poudeoly] JEON “LN ‘opqesouyn,A ‘AA ‘polosuepug ‘Ny ‘porosuepugq

AT[POIID “UD (SLOT Te 19 WNoouvjog-soesop[) SNCS UOIVAIBSUOD ULIQUIO][OD :jSI'T POY [CUOTIEN :SNjBJS UOIVALISUOD [BUOHBN ‘IQUIO[OD UI poyodal UOTJeADTA WINWIXe/] sad] | *XBIA

(Vd) OHOC ‘sseodde 31 s10Jo19Y} “UIseg DYIdeg dU} UI SINDDO ODOYD Jo JUOWIedap oY} UI sppdi DiIUOJaYD SaIdads dy} 1OF APATOW SUNSON ‘(QTVWO) OHO ‘sivodde 1 s10jos9y} “ulseq uvaqqlie)

dY} UI SINDDO Qd0YD Jo JUusUIedOp OY} Ul YJDILAGuA SAJaYIOWJALY SHIOAS dy} JOJ APATOW SUTJSON “00} S19Y} S}SOU JI SUBS (q) PUB “UBdD0 9} UT dOUdIINDDO ATUO SUB (VY) *,.UOTNGIISIG

[eotfodoany,, UUIN[OS UT Soj}IN} vas JO] ‘(TA) epeysiA pue (VA) sednea “(OVA) Boned Jap aTBA (TOL) PUITOL (ONS) eJ9Ng “(NVS) Jopuejues “(qVvs) euljeyeD Bug A vIDUSpIAOI “SoIPUY

ues “(STX) epreszesry (INO) ofpurnd “(LAd) oseumnyng “CVSN) Jopuejues op OJON “CYVN) OULRN “CLA BIO (COVIN) PusTepseyy “(OVT) elfen eT (INH) e1InH “(AND) srerAeny

‘(VND) Brurenyd “(NMD) eoseweurlpuny “YOD) &qops9D “(OHD) 92904D “(SAD) Jesap “(AVD) Boned “(SyD) seuRseD (OVD) BlenbeD (TVD) sepred (AO) Boeh0g “(TO) Jearfog (TLV)

oonuepy “(VaV) eonesly “CLNV) einbonuy “CvIV) seuozeury :(quounedog Aq) uoRNQLysIp [eayOd0ay ‘(dyI9eq) Vd Pue (ODOUTIO) YO “(eoneD-eusjepseW) VO-OVIN ‘(uRsqqueD) VO

‘(UOZEUIY) VIN'V “JUasoIdo1 SULIOJ POUdLOYS OY} ‘sasvUIBIpP IYdBASOAPAY Ul UOTNGLYSIG, SUOTLIADIQGY VIQUIOJOD JO Sd]}IN} dy} JO SNje}s UOTJeAIDSUOD pUe “UOTNQLYSIP ‘AJISIOAIG “[ IQey

(4 ¢

April 2022 | Volume 16 | Number 1 | e306

TAZ

Amphib. Reptile Conserv.

t al.

aeze

-LAd

998L Y-HAVI :CIA

'8EL1 ddY-NHW-NOI ‘NVA ‘LEZ [enb-s TW

‘0€L1 dey-NHIN-NOI ‘LAW ‘SZL1 Y-HAVI

‘AND ‘80670T6L/SUOHTRAJOSqO/810 JSITeINJeUT

mmay/sdny VND ‘91 EL dey-NHIN-NOI ‘SVO

“PSI lenb-S TW ?OVO ‘O€LE U-HAVI ?VINV

VI7T81 LO-HAVI

“CIA ‘6PL1 U-HAVI ONS *LS8Z U-HAVI :NVS

“68 UO fenb-S TW “LAW “L090 Y-NHW-SIN

OVI -£6199T19/SUoTeAJasgo/S10 'jsIyemnyeur

MMM//-SANY IQH'€8PSPS€3/SUONBAIISG 0/310

‘ystpeInyeuTMMAM//:SdyY ‘OV’ “6€L1 Y-HAVI

“YOO ‘9191 U-HAVI ‘OHO ‘8¢z anb-s TAL

‘SAO ‘¢¢z [anb-S TW :SVO “pr9L U-HAVI ‘AOP

“CELT U-HAVI ‘10d -9SpZ1T WNSN ‘TLV

‘Q09L U-HAVI :VUV ‘PL688EST/suoneArlasgo

/S10'ystTeINVeur eIquIO]09//:sdyYy - NV

IT fenb-g TA :d VS

TPEET U-VNHW :CIA “L8€9-Y-HAVI

‘ONS “PLSL Y-HAVI “LAd “€8S H-dNNOW

“VSN :0PL80 U-HAVI ‘LOW 10000 dau

OVNNED ‘OVI ‘PS8¢ U-HAVI ‘OV T “ILSL

U-HAVI VND “PETLI U-VOHW :WOO “19PZ 11

HNWN ‘SHO “17S PL I [/suoneArasqo/310

“yst[eInyeut eIqwuo]o9//:sdyYy -SWOD °L9SL U-HAVI

‘OVO ‘€69 Lda U-fNdW ‘TVO ‘68€9 U-HAVI

‘TO"dU ‘OOLT I 66¢/SUOIVAIOSqO/B10}SI[eINyeUl

“erquiojoo//:sdny “TLV “perl U-HAVI -VuUV

‘PIOLI U-VONHW :LNV ‘791697 HOM: VV

98091 O-AN ‘OVA

‘TLEST U-WNHW “TOL:90L707L/suonearasqo

/s1oTsIperNjeUT MMM //-SayY ‘ONS “pTEE U-NHWN

-SIN (NVS “IIS Ldad-fndW ‘Sta ‘7ez ONUV

‘INO ‘svz lenb-S TW -UVN “SZ7EI8 HNN

“LAW ‘7Z8L NOI ‘OVW ‘I8€I1Z 4I9H-ZAW

‘INH ‘6p0¢ do¥-NHW-NOI ‘NNO ‘ZS9ZI

YWVNHW YOO ‘¢h01 H-HOOOZT100 ‘OHO

“pS960I1 U-HNWNV ‘NVO ‘1971 Lda-fndN

“TVO ‘p877 U-NHW-SIN ‘AO “€7581

‘

UWVOHW ‘TOP -887ZT Y-VNHWN -LNV

bO8thp HNWA ‘OHO

9091 Y-H4AVI ‘OHO

s1aqono,

6107

‘Je 19 SOLIOL-OouRT ‘TOT [e 18 VIPUDITV

-ALIDADYIA SOOT ‘[e 19 ZOINWIeY-SeBICA,

610¢ BAPUSITV

-AUOAOYIT “OLOT Te 10 OND-eyedeZ “C107 ‘Te

19 PIO[-OULISLD “PIOT ‘Je 10 BALIOD-Sa}UOJ]

‘PLOT PHOIS-BLIeSUY ‘TOT ‘Je 10 BIDIVH

-O89][2D ‘1107 [odueY-eUIPoW ‘O1OT “1219

ZOMUCY-SCBILA “POOT ‘[e 19 ZOUIOH-zouoNq

‘I86I Sojesny-o8n] pue elopl-ourjsed

QS 107 ‘Te 19 PUIPa||-O1210,J

‘TLOT BIOP-OURISeD pue vUIPapy-O1010,4

6107 O[[0805-TelBareD “L107 Te

0 ZOUNIeP[-SOI9908D “L107 ‘PILOT ‘Je 19 vAMOD

-SOUOP SOOT SBBIVA-BUNDY “ZI OT [e319

ALO ?1 107 [SURY-PUIPO! “ZOOT ‘Te 10 ZR

6107

O]]0300-yeleareD “3107 [ev 19 OULZOT-OSULIY

“PLOT ‘TP 10 BIOINJA|-ZonsLIpOY “GE [OT “OT LOT

[B19 OPTeAIH “goOT seUul[eS-sesieA pur

RIDLOH-ONSCD ‘POOT ‘[e 19 BIOP[-OURISeD

ecl0c [e198

BUIPIA[-O19IO-] STLOT [e 39 OUDIOJ[-BLIOUDY

LIOT ‘Je 10 ZOU Y-SeBICA,

“PLOT Publ puk SOT[eQSD “GZ 107 ‘TR 19 YoY

UONQLYSIP IYAvsAGOIS 10} S9IUIAIJOY

a 00S‘I

| OOL‘T

b cal loa OVA ‘OHO

a OHO ‘LNV Ty dy.

snqe}s ({se

WOIeAasu0d sTUIapuy Wi) ‘AIT

euoneN Xe

CIA

‘NVA “LAd ‘LAW

‘AND ‘WND ‘SVO

‘OVO ‘VUV ‘VV

CIA

“TOL ‘ONS ‘NVS

‘LOW “OVW ‘DVT

‘INH ‘VND ‘OO

‘OHO ‘SHO ‘SVO

‘OVO ‘AOE “10d

“TLV “VuUV “LNV

CIA ‘ONS “LNd

“VSN (LOW “OVI

‘OV 1 ‘VND ‘OO

“SdO ‘SVO ‘OVO

“TVO “10d “TLV

“WUV SLNV ‘VV

OVA “TOL ‘ONS

‘NVS ‘STa ‘INO

AVN “LAW “OVI

‘OV ‘INH ‘NNO

“YOO “OHO “SHO

“‘NVO “TIVO ‘AO

“TOd “TLV “LNV

worNgiaysip

[eontjod0a

(10119)

VSN PIUISIIA

“TZ WoTIaT[OO

19D 9d WUN

‘99L| snoeuury

eOLIOUy

YINOS ‘payeso]

ION “pZ8I xIdg

88Z1 Yosieg

eory

BISOD ‘asor URS

‘09L1 NHNW

‘OL8 1 “Hnooog

pue [lIguinqg

weuUlIns

poyes0] 10N

eyeurayeny

ud10d [4

“eqloeuns ()

Ory “L806 'PIIZ

NHNW ‘IS81

[lauinq pue

xX xX xX ‘uolqrg “[awing

BIquO]OD

x x

“Q00YD “OIIeZIg

‘p08Tr HNINA

“LP6 1 IPIWUYDS

LS8I Zissesy

RIquUIO[OD

“od0YQ ‘orons

ory “9091

U-HAVI ‘L107

ZU pue

‘soy[eqay “aTTeA.

[ep “Y-sesie,,

Aypesoy adA}

pure ‘adAjojoy

Toyny

Da [oo [aoa Pano Poa

sosvurvip s1ydeiso1pAy Ul WOINQLySIG

(seyJuewy

seyeg op AOdOLIOJ] ‘OSIO}IOL,

Po}OOJ-MOTIOA URITIOUTY INOS)

snypnayuap sIplouojay,) ‘Z|

(Oooo ‘seloy seed

ap AOSOLOJ {ASTOLIOT, P9}00J

POY) SNLUDUOGADI SIPIOUOJaYD ‘| |

oeplUIpNysay, Ape]

April 2022 | Volume 16 | Number 1 | e306

(eyueMs ‘opin,

pnyl poweos-ay1y A) aipjnsoqgjp

sapoidioos uoudajsOUuly

113

(IgiyD “ojnoedey,

‘opainy pnyy uords0ss) sapoid.ioas

Saploid4s0Is UOUAAJSOULY ‘(|

(edeyng

onoedey ‘opin, pny peddiy

-OUIY MA UOYINOS) ayouinsurjsod

WUWNUOJSOINA] UOUAAJSOULY 6

‘

(OZOI], op ezaqeg ‘aN, pnyy

S$ uun(]) MuNnp UoUsa]SOULY 8

oeplusgjsoury Ape

(oyeNY [Op La}OdIH “JOpl[s ory)

‘AOU ‘ds 1luapau SAUMaYIDAL

(vIquIO[OD UI pasn suv puL

Ysi[suy Ul oWeU UOWIUIOD) UOXxey,

Amphib. Reptile Conserv.

distribution, and conservation

Turtles of Colombia: diversity,

70901 M-IHONIS ‘CIA

‘96€L U-HAVI (NVA ‘P6EL U-HAVI “LAW “L6EL

YU-HAVI ‘AND ‘L6€L dea-NHW-NOI -VND

“8r1 1enb-S TW ‘OVO “6Z1 [enb-s TW :VINV

P8E9-U-HAVI (ONS -689LEL0P/SUONeAIOSgo

/310 71sI[eINyeur'eIquiojos//:sdyy -QOVIA

“89ELI NOI ‘SHO “P6S ITLPE/SUONeAIOSgO

/810 '7jsI]eINyeUT VIQUIO[O9//:sdyY : TOW

SSL8U-HAVI

CIA ‘9PL8-U-HAVI SLAW “TTEL NOL: VaVv

OOLL U-HAVI ‘NVA ‘OLLI Y-HAVI .Ld “£921

WHAVI AND ‘O0LL NOI :WND ‘SOEL U-HAVI

‘SVO ‘VOLT NOT:OVO “7921 U-HAVI -VNV

‘

8h LO-HAVI -‘OVT -TTOZT VOHW ‘OHO

000L1T Y-VONHW -OHDO -cEEOl LO-HAVI -NVO

Svcy LO-HAVI -OVT -CIE01 LO-HAVI -NVO

096r U-S£D ‘ONS “SSZy LO-HAVI DVT

“ELEOL LO-HAVI (NWO “1676 LO-HAVI “10d

‘9876 LO-HAVI ‘OWW “ISTP LO-HAVI ‘DVT

S1ayINOA

O70 ‘Te 19

OSSP’'T SOT OT OSSE'T pur JINOOULJOg-soTLIO/

6107 110809

-[elearea “pIOT ‘Je 18 BALIOD-SaUOJ “9S [OT

‘TLOT ‘TB 19 BUIPA[-O1OI0,] °] [OT JosuLY

-BUIPSL “9TOOT Wpsp| puB eIOP[-OURISeD

IZOT Je 18 BYUND ‘OZOT ‘[e 19 ZOIMWUIBY-SeBILA,

0c0c Te

0 ZOMULY-SESICA ‘PIOT PLOIS-eWIeSUY

‘dZ10T OSSe’'T puke yINOOURIOg-SoeIOP[ “1 1OT

‘JR 19 OLEJTV “8O0T PIEYIUd 6961 Wpoy|

“070

OpueqQ-ZopuRLLISH “9107 OLE “9107

‘TR 19 ZAWUIOH-VIOATY SS TOT ‘Te 19 OaTTeH

-ZoIUeYy “plOT Te 19 IeuNpy-OUsIOW “TOT

‘Je 19 O1DIIO “3007 ‘[e 19 ZoUTIRI-OUNeg

‘[ssolg uy]

TCOT ‘TP 19 ZOUNPY-sojuaLUEg [TO ZouSUUIL

-SOLUBPY pUk SO[GOY-VIOATY “6 10T MOY

pure WOyq “Qo 1OT ‘Te 10 ZouNP|-soyorueEg

‘PLOT Te 39 ZoUNP[-soyaLUV “gQQz Te 12

vionbsopl-oyoeuley ‘766 ‘Je 19 OYDoIOWY

[ssorg uy]

TCOT ‘TP 19 ZOUNP[-soJUATLLg “OZOT ‘TB 18

O]]lued-zonbsea “eQz7Oz ZoUN/|-SsowsLUeg

pue osaljen-zoruey “q¢ [07 [ev 19 Zoe

“6107 Hoxo_ pue Woydq “9107 OUR “p1OT

‘ye 19 IeUNP[-OUDIOP[ “E1OT Te 19 ZOUN|Y

-SOJUdILUeY ‘3Q0T BUIOY pue OYDOIOUTYy

[ssorg uJ] TZOT ‘Te 19 ZoUN]

-SoJUdIUeY “[ZOT SowWsLUeg pue osa]]/eH

-ZONWCY [TOT |e 18 ZONSLIPOY-ZolealTy

“GOTOT “BOTOT ZOUN\-soJuaLeg pue

Osa] [eH-zZoNWIeY “OTOT ‘Te 18 AqlI)-seueD

6107 HOyoY pue Woy “O07 O}oUeY

“BC TOT Te 39 ZOUNPI-sOJUATUL “p[OT Te 12

seLIy-O][Iniy “plow ‘[e 39 JeUNy-OUdIO/I

0c0T

opueqa-zopueuloH “eOzOT ZOUN-sojUSTLUEg

pue o8aljen-zomuey “6107 ‘Te 19 epeouoyy

‘LIOT ZopueuIeJ-zopuPuOH pue esourdsq

~OOURLY] “BC LOT [B19 Z9Rd “PLOT [2819

IeUNJ[-OUdIO| “€ [OT ZOPULUID.J-ZOPUPULIDH]

pue eso,1Q-zoueP ‘ET OT ‘Je 19 O1O.LOg

UOHNQALYSIP IYdv1G0IS 10} S9IUIAIJFOY

snqvjs

UOIVA.AISUOD

[euoneN

awopuq

UWI) “AIT

“XB

CIA

‘AVA “LOd ‘LAW

‘AND ‘VND ‘SVO

OVO ‘VUV ‘VV

‘

ons ‘DVN Yoo

“SO “TO” “TLV

CIA ‘LAW “VuV

NVA

“‘LNd ‘AND ‘VND

‘SVO ‘OVO ‘VV

OVW DVI ‘OHO

“LNV (d) ‘ONS ‘VS

VN ‘OVI ‘OVI

“YOO ‘OHO ‘NVO

“TOd “LNV (V)

OVA

YIVN ‘OHO ‘NVO

(dQ) “OVA ‘UVN

‘OHO ‘NVO (Vv)

dvS DVN

‘(Wd) OHO “INV

(@) “OVA ‘ONS ‘AVS

VN ‘OVIN ‘DVT

‘YOO ‘OHO ‘NVO

“TOd ‘LNV (V)

ons ‘dvs

‘OVW ‘DVT (V9)

OHO “TO” “LNV

(@) ‘OVA ‘ONS AVS

VN ‘OVI ‘DVT

‘YOO ‘OHO ‘NVO

“TO” ‘INV (V)

dvs

‘OVI ‘DVI (a)

‘ONS ‘AVS ‘OVI

‘OVI “10d (Vv)

woHNg!ysIp

[earodoa5

sosvulevip sydeiso1pAy Ul UOINGLysIg

BOLIOUY

yinog “9¢Z8

NHNW ‘7181

JOBSIOMYIS

RIQUIO[OD “aINNS

* OfataoulsS

Jo APUIIA,,

086SZ HNAA

“8S6] Wapo|

pue jiosuez

‘

RIQUIO]OD

“epeysiA eg

Ory “SSL8

U-HAVI “0707

‘[e 19 Y-sesIe,,

euRINy

Yyouary “poyesoy

jou adAojoH,

“ESI Joplouyps

seas o1eLIpy pue

ULOULLIOIPS/|

‘posoquinuun

dNWZ “19L1

T[epueA,

Ep8I JOBUIZI

sourddityd “Aeg

BpuRYl “LZ ul

A]qissog “6781

Z\[OYOSYOS A

puejs]

UOIDUDNSV ‘976

WAN odAquss

“SSL snoeuur’y

spuvys] epnuriog

‘O€T NNWZ

“99L| snoeuul’y

spur|s]

UvOLIOUTy

poyeusisop suON

“SSL snoeuur’y

1181 Teddo

Ayrpesoy ad A}

pue ‘adAjojoy

“Ioyny

(eZLIND “epuoIpoH ‘apn,

eqqID) Yqqis sduuuajzosayy {TZ

(e1oURUO|Y

“euIgoURLIeD ‘opin, popesy-peoy,

S,1yeq) "yp skunuajoosay ‘OT

(OD0ULIO [Op

RILUILLI| ‘QOUIDOUTIO) BIULL

“OLN, BJLUIL}LU ODOULIO)

‘AOU ‘dS sisuadoUl.1o SNJAYD ‘6

(seuozeury

[op elewRePy fopNy, eyeureyeur

UOZLULY) DIDLIQUIS SNJaYD °3]

aepHeyD Aprurey

vAIPOANI[g UIporqns

(pney ‘vued “opin, yoeqsoyyes])

DaIDIAOI SAJAYIOWLAC] *L |

aeplApaypourog Ape

(euyjoy ‘opin Ao[prey

dAT[O) Daavaljo sdjayoopidaT ‘9 |

(vouvlg BSNWO], ‘Ops, BSNIOL,

QTUN I], UsIH) sppdiu piuojayy *S |

(Aored ‘op []}GsyMeH)

DIDALAQUIL SAJAYIOUNJAAT “P|

(euozaqey “euense, ‘opin,

peoylesso]) YUJalvo DJJaIDD *€|

oepruopay,) Ape]

(vIquIO[OD UT pasn sue puL

Ysi[suy Ul ouvU UOWUIOD) WOXE],

April 2022 | Volume 16 | Number 1 | e306

114

Amphib. Reptile Conserv.

Paez et al.

Leeel Y-VOHW :CIA -OLT H-ONHW

.LAIN °99 [2nb-S TI ‘SVO ‘Z6LE U-HAVI ?VUV

6€fEl Y-VONHW

“GIA “807 [enb-S TW “Ld ‘707 H-ANHW

SLAW ‘SLTEVP6 [ /SUOIJAINSGO/10"7}SI]eIN}eUT

“erquuojoo//:sdny ‘AND ‘Or81 Y-HAVI ‘OVO

“SPEL6 WHNNV :VUV “PZ0LT U-VAHW VV

98LL doYy-NHI-NOI ‘VINNY

O8S801 WNSN ‘TOL

086€ U-HAVI ‘ONS ‘77681 U-VNHW :NVS

‘SOOLT U-VNHW ‘OVW -096€S6r9/SUOHIeAIASqo

/si10 ysTTeINyeUT MMM //SCHIY “IH “88S8¢ 1

HN ‘NNO ‘76E8T U-VAHW “WOO “77

YHAVI ?TVO ‘LETLT U-VOAHW ‘TOE

‘Epers ANS ‘TLV °606L1 U-VNHW :LNV

L68L1 Y-VOHN

‘CIA “PSL Ldata-fndW LAW ‘8S6r U-HAVI

“SVO ‘0982 deYy-NHW-NOI ‘OVO ‘0st Y-2ON

-NHW ‘VUV ‘0082 dau-NHWCNO! ?VINV

PCPL U-HAVI ‘CIA ‘7861 dou-NHW-NOI ?VND

ETL U-HAVI :CIA ‘0662 d°Y-NHW-NOI 'VAD

ILLL doY-NHW-NOI ‘CIA ‘06ST U-HAVI ‘AVA

‘TOTS U-HAVI “LAW “6079 U-HAVI (AND “PLL

doy-NHW-ONI ‘WMD “€€8t U-HAVI -OVO

LL9T U-THONIS :-NVA

ZSOI

Ldate-fndW ‘AVA “O€1¥7Z WNSN -LAds«

“OOrrPO90P/SUOTIeAIOSqO/810jSIpeINjeUl “RIQUUIO[OO

/-sdny ‘AND “L8€01 U-IHONIS :WND “18

fenb-TSW ‘OVO °9Z9L d2a-NHIW-NOI “VV

PI9L deY-NHW-NOI ‘VV «

S1ayINOA

OZOT ‘Te 38 OINBIS

-epoatndas “¢ [07 ‘[e 19 PLIOIS-eILIeSUY

“PTLOT ‘Te 19 MODULIJOg-SoTeIOPI “600T ‘Te 19

os uTy “YZOOT Wepsy] pue eIOPY-oURIseD

QS10T ‘[e 18 MNoouejog-soelojy

“PLOT SIATRD-eIsooV puke ojifusy “€ 107 [2 10

RIIDIS-VILIVBUY “TOT Je 19 BUOTeOSY “8ZNOT

Wapojl puke PIOJ[-OURISED ‘go6] Wapoy|

QZ10Z

‘Te 19 SOTTEqID “SOOT BORD *g96] WOpa

6107

O]]O30_-[efeareD “3 [OT PIOP-BZOpUd|y pure

sepyseg-odensoy ‘¢ 10% ZRd “€ LOT “AT 107

‘JR 19 Zed *LLOT [OSueY-eUIPa| *ZOOT ‘Te

Jd Zoe “WTOOT Wpoy] pue BIOP[-OURISeD

8107 ‘Te 19 OFY [AP “S TOT ‘Te 19 selayyeD

“ZOUIURIAL “BZ [OT “JB 19 SO[TEQSD ‘JTOOT

Wopeyl PUB BIOP[-OURISeD “696 WOpe/

2S [OT OSSL'T pure JINOOUBJOg-SoTBIOW] <T1OT

‘Je 19 preytulag “6007 ‘Te 19 oJtluay “E007

‘Je Jo BIOP[-OURISeD SOZNOT LIOP[-OULISeD

QZ107 ‘Te 19 BSSO BT 9-096] WPS

ITIOT Te 19 WNosuKj}Og-soyeJOY]

QZIOT Te 19 HNoouRjog

~SOTPIOY “OTOOT Wopoy\] pue eIOP[-OURISeD)

Tc0c OMLL

eC LOT [2 19 BSSE BT 9] “E86 Wps||

6107 ‘Te 39 BYUND ‘ZI OT Je 19 BUTIOW

PTIOT OSSe’] pue WNosURIOg

-SO|LIOW <PTOOT Wdpa| puke LIOP[-OURISeD

UOHNQALYSIP IYdv1G0IS 10} S9IUIAIJOY

snje}s

UOIVA.TISUOD

jeuonen

awepuq

([sv

Ul) “AIT

“XB

CIA ‘LUN

AND ‘SVO ‘VUV

CIA

‘AVA “LOd ‘LAW

‘AND ‘VND ‘SVO

‘OVO ‘VUV ‘VINV

NVA

“LOd ‘OVO ‘VINV

TOL

‘ONS “NVS ‘OVIN

‘INH “NNO ‘YOO

‘SAO “TVO ‘AOE

“TOP “TLV “LNV

CIA ‘NVA ‘LNd

“LOW “VND “SVD

‘OVO ‘VUV ‘VV

CIA ‘AND ‘WND

CIA ‘NVA ‘LNd

“LAW ‘AND “VOD

‘OVO ‘VUV ‘VV

CIA ‘NVA ‘LNd

“LAW ‘AND ‘VND

“SV ‘OVO ‘VV

NVA “WND ‘VV

CIA ‘NVA

‘LAd SLAW ‘AND

VND ‘OVO ‘VV

‘

66NVA “LAd

VND ‘OVO ‘VV

‘

woHNg!ysIp

[ear}odoay

sosvulevip 1ydeiso1pAy Ul UOINGLysIq

elONZoUdA,

“‘seulieg opelsq

8C/8CI WSZ

“SE6I JOIN

euRANd UI SIOATY

‘Trl ANZ

“P81 [PYSOLL

ROLIOUTY YNOS

‘TOATY UOZeUTy

so] odAQojoH,

‘6r8 | BIfeUIOD

eBIquiojoD

“gj030g Op 24

ByURS “C068

NHNW ‘Zs81

juguing

POLIOUY

(NOS “L66L

NHNW ‘7181

JOBSIOMYIS

[Ize1g “SOOUT[OS

ory “O/LIST

SINZ ‘7781 xIdg

POLIOUY

YInos ‘jsoy

od JOO “ZI81

JOBSIOMYIS

6981 edop

Izeig

erpselg “L1V6

NHNW ‘7181

JOBSIOMYIS

0/900€

WSZ ‘781 xids

dog ‘seuozeury

“Rayl[eH) Jo

AVIUISIA,,

“QEl vec WNSN

“p86 IsWIY

eueiny

yousry ‘ouuared

“poyeso] jou

od MOJOH “Z6L1

Joprouyos

voloury yyNoS

‘vLT7L 9P61

SININHN

“6961 SUSHIOIAL

[Izelq

“ered ‘(9d4}0399])

cove Lrel

NHN

‘9¢8 1 ABIQ

Aytpesoy od 44

pue ‘adAjojoy

“oyny

(eroueges “orauRqes

osedeyen ‘oyun |, payoou-apis

BUURARS) 1/804 SIWMaUOpOd '€¢

(eAv9919 |,

‘KBOOIOL, ‘apn, JOANY panods

-MOTJOA) SYLfiun smuaUus0pog ‘TE

(ostdng

SOPUN], JOANY poposoqny-x1s)

DID[NIAAGNI]Xas SIUAUIOpOd “| E

(duapppsnyy Ory [op

VSNYOL, ‘OPIN JOArY eusyepsey|)

pupdma] SMUaUr0pOg ‘(E

(nelly

‘edesrey ‘ayn, JoATY ueoLIowy

ynos) vsuvdxa siuaus0pod ‘67

(andry9 ‘ondryD

‘OTN, JOATY popeoy-poy)

pypydas0dyjAsa SIMAUIOPOY ‘8T

(uozaqeg “opnzeqea

‘OPN, JOATY uozeury popeoy-31g)

snupyliauinp snjoydasojjad ‘LT

aepeplMmaud0pog Ape]

(eyeueyeyy ‘oredoy,

SOPIN, poyoou-aplg s,AoJooo)

snuvosffoas sdouday dg ‘9T

(a191yoV

‘efoy esnqoy ‘apn, poyoou

-OPIs poy) sadifia sduauryy SZ

(endesreyD

‘edeseyD ‘apn, poysou-poysim |,

poyoeg-yovlg) viouounjau ‘dg

(endesreyD

‘edereyD ‘apuny, poyoou

-paysIm, Wlayseq) vjpydaatjnjd

pypydaadjnjd shay] d “pT

(9]IN], popeoy-pso], s YINULIA\ )

14jnudan sduuuajoo0sayy “ET

(uozaqey ‘odes ap ezaqea

‘OPIN, popeoy-peoy uozeury)

sdaoiupd SMUUAIOSa ‘TT

(vIquIO[OD UI pesn suv puL

SIsuy Ul BWIeU UOWUWIOD) UOXxe],

April 2022 | Volume 16 | Number 1 | e306

115

Amphib. Reptile Conserv.

Turtles of Colombia: diversity, distribution, and conservation

on both the Pacific and Caribbean coasts of Colombia,

describing the Caribbean presence as “accidental”

(Ceballos-Fonseca 2004). However, this species has only

been documented to occur and nest along the Pacific coast

of Colombia thus far (Barrientos-Mufioz et al. 2014;

Barrientos-Mufioz et al. 2015b), with no documentation

of the species occurring or nesting on Colombia’s

Caribbean coast. It is possible that individuals may

occasionally traverse the Caribbean waters of Colombia,

as sightings in the Caribbean and sporadic nesting on

Caribbean islands have been reported (Eckert and Eckert

2019; Moncada and Romero 2015), but the principal

nesting colonies of this species in the western Atlantic

Ocean occur in Guyana, Suriname, and French Guiana

(Eckert and Eckert 2019; Marquez 1990).

Phrynops tuberosus (Peters, 1870). Historically, the most

widespread chelid species in South America was considered

to be Phrynops geoffroanus, withthe morphological variation

throughout its range leading some authors to consider it as

comprised of different subspecies, including P. g. tuberosus

(Muller 1939; Wermuth and Mertens 1961; Duellman

1978). The taxonomic revision of Phrynops by McCord et

al. (2001) elevated P. tuberosus to the species level, and it

is currently considered to be restricted to northeastern South

America in Venezuela, Guyana, and Brazil (TTWG 2021).

However, Ferrara, et al. (2017) claimed that P tuberosus

occurs throughout the northern Amazon of Ecuador, Peru,

Colombia, Brazil, and Venezuela, based upon the results

of a molecular analysis in the thesis of Carvalho (2016).

While recent publications from Brazil have shown that the

Phrynops complex is comprised of several cryptic species,

or at least evolutionarily significant units (Friol 2014;

Carvalho et al. 2016), the molecular evidence arguing that

Colombian populations should be considered P. tuberosus

rather than P. geoffroanus has yet to be published, so we

therefore do not replace P. geoffroanus with P. tuberosus in

this checklist.

Podocnemis lewyana (Duméril, 1852). Castafio-Mora

and Medem (2002a) reported that this species had

been extirpated from the Rio Rancheria (La Guajira

Department) based on a mention of this conclusion in

a non-peer reviewed document by Hurtado-Septlveda

(1973), and this claim was perpetuated in later literature

(i.e., Paez et al. 2012a, 2013). However, a niche

modeling analysis to predict potential habitat for this

species both now and under different scenarios of future

global climate change (Ortiz-Yusty et al. 2014) failed to

predict the presence of P. Jewyana in the Rio Rancheria,

and the visits to this drainage that were part of the

ground-confirmation effort in this analysis also failed to

detect this species or any indications that local people

recognized it from photographs. They concluded that the

report by Hurtado-Septlveda (1973) of its extirpation

from Rio Rancheria was questionable, and for this reason

we do not include the La Guajira Department in the

Amphib. Reptile Conserv.

distribution of this species.

Recent taxonomic changes

Chelidae

Chelus. Matamata turtles exhibit geographic variation

in carapace shape and color, with individuals from

the Orinoco drainage having rounder, lighter colored

carapaces than Amazonian individuals (Pritchard and

Trebbau 1984; Sanchez-Villagra et al. 1995; Pritchard

2008). An examination of two mtDNA fragments (Lasso

et al. 2018) also revealed haplotype differences between

the Orinoco and Amazonian individuals from Colombia.

Finally, an examination of three mtDNA fragments,

one nuclear DNA fragment, and multiple SNPs from

individuals across the range of Chelus fimbriata (sensu

lato) revealed a deep phylogenetic division between

samples from the Orinoco, Rio Negro, and Essequibo

drainages versus samples from the Amazon and Mahury

drainages, prompting Vargas-Ramirez et al. (2020) to

elevate the former clade to the species level as Chelus

orinocens!s.

Mesoclemmys heliostemma (McCord, Jospeh-Ouni,

and Lamar, 2001). This species was described based on

five voucher specimens and nine live individuals from the

western Amazon region, specifically northeastern Peru,

eastern Ecuador, and southern Venezuela (McCord et al.

2001). Molina et al. (2012) examined eight additional

individuals from eastern Peru and northern Brazil, and

concluded that the species is valid and morphologically

distinct from M. raniceps (but see Cunha et al. 2019).

They suggested that reports of M heliostemma for

Colombia were cases of misidentification of M. raniceps

individuals. The previous TTWG checklist (TTWG

2017) mentioned Colombia as likely to include M.

heliostemma, presenting a range map with a polygon that

included Colombia but without any point locations. Our

searches failed to find any vouchers or literature reports

corroborating the occurrence of this species in Colombia.

Finally, Cuhna et al. (2019) reviewed the convoluted

history of the taxonomy of the genus Mesoclemmys and

presented evidence indicating that females of MZ. raniceps

may oviposit clutches that produce hatchlings with both

M. raniceps and M. heliostemma phenotypes. They

concluded that M. heliostemma should be considered

a junior synonym to M. raniceps and warned against

describing species solely on the basis of differences in

color patterns. This recent taxonomic proposal, combined

with the lack of any vouchers or literature records for the

occurrence of turtles exhibiting the “M. heliostemma”

phenotype in Colombia, led us to exclude this species

from our checklist.

Mesoclemmys ranicpes and M. wermuthi. Cuhna

et al. (2019) not only synonymized M. raniceps and

M. heliostemma, they also resurrected the species

April 2022 | Volume 16 | Number 1 | e306

Paez et al.

Mesoclemmys wermuthi, which had been previously

synonymized with M. raniceps (Bour and Pauler 1987).

They also argued that the name Mesoclemmys maculata

had precedence as the correct name for this resurrected

species. Although the most recent TTWG (2021) checklist

chose to recognize the species, it retained the name M.

wermuthi. Apparently both M. raniceps and M. wermuthi

have been reported to occur along the Colombian borders

with Peru and Brazil (TTWG 2021), but our searches

only produced one voucher specimen identified as M

raniceps. Thus, this is the one instance in which we failed

to find a rigorous record for a species purported to occur

in Colombia (M wermuthi), or alternately, the voucher

we located is actually a specimen of M. wermuthi that

was misidentified, in which case we lack a rigorous

record for M. raniceps.

Emydidae

Slider turtles. The slider turtles in the Atrato River

drainage and the Gulf of Uraba region in northwestern

Colombia have long been recognized as morphologically

distinct from the slider turtles from other more eastern

populations in Colombia, located along the Caribbean

coast and in the Sinu, Magdalena, and the lower

Cauca river drainages (Williams 1956; Medem 1958).

Medem (1962) and Ceballos-Fonseca and Brand (2014)

summarized the details of these morphological differences

which involve plastron and color pattern characteristics.

However, over the past two decades, the taxonomy of

both “western” and “eastern” Colombian slider taxa has

been unstable (as has the taxonomy of slider turtles in

the Americas overall). The names assigned to the more

widespread eastern slider turtle taxon in Colombia

include Pseudemys scripta ornata (Williams 1956),

Pseudemys scripta callirostris (Moll and Legler 1971;

Pritchard and Trebbau 1984), Trachemys callirostris

(Seidel 2002), and Trachemys venusta callirostris (Fritz

et al. 2012; Parham et al. 2015). Similarly, the names

employed for the western Colombian slider turtle taxon

include Pseudemys scripta ornata (Williams 1956),

Pseudemys scripta venusta (Moll and Legler 1971),

Pseudemys scripta ca. venusta (Pritchard and Trebbau

1984), Trachemys venusta (Seidel 2002), Trachemys

venusta uhrigi (McCord et al. 2010), and Trachemys

medemi (Vargas-Ramirez et al. 2017).

Studies on the relationships of these two taxa to

other slider turtle species and subspecies, as well as

phylogeographic studies of their origins, have also been

equivocal. Over the past two decades, various cladistic

analyses have concluded that the two slider turtle taxa in

Colombia are either closely (Stephens and Weins 2003) or

distantly (Seidel 2002) related, and are of Mesoamerican

(Jackson et al. 2008; Fritz et al. 2012) or Caribbean

(Stephens and Weins 2003) origin, or both (Seidel 2002).

Most recently, Vargas-Ramirez et al. (2017) expanded

upon the study by Fritz et al. (2012) by adding samples

Amphib. Reptile Conserv.

from 12 individuals of the western Colombian slider

turtle to their genetic analysis of mtDNA and nuclear

DNA. They concluded that South America has been

colonized twice by slider turtles from Central America;

first by the ancestor of Trachemys dorbigni (currently

occurring in Brazil, Uruguay, and Argentina) and the

western Colombian slider, which they elevated to the

species level, assigning the name 7rachemys medemi.

Much later, Colombia was again colonized from Central

America by the ancestor of the eastern Colombian slider

(Trachemys venusta callirostris) and the Venezuelan

slider (Trachemys venusta chichiriviche).

Translocated native and exotic species

The occurrence of Chelonoidis carbonarius on the

Caribbean island of Providencia (CAR macro-drainage,

San Andres, Providencia, and Santa Catalina Department)

has long been assumed to be due to either pre-colonial

or more recent human transport (Castafio-Mora and

Lugo-Rugeles 1981). This also seems to be the case for

the population of Kinosteron scorpioides albogulare

occupying the island of San Andrés (Montes-Correa et

al. 2017; McCraine 2018). Medem (1969) also reported

several apparently successful attempts by colonists

in the Amazonian region to introduce populations of

Podocnemis expansa into the upper Caqueta and Caguan

rivers (AMA macro-drainage, Caqueta Department),

presumably because of the economic importance of this

species, but we failed to find any museum vouchers to

support this claim. The only other documented case to

date of apparent artificial range expansion for a turtle

species in Colombia is from a publication (including

museum voucher information) documenting — the

occurrence of Trachemys venusta callirostris individuals

in several locations 1n Quindio Department (Cordillera

Central of the Andes mountains) at approximately 1,500

m asl (Adames-Jiménez et al. 2018).

Our searches for voucher specimens in this study

failed to document any additional suspicious location

records for Colombian turtles (i.e., individuals collected

far outside of their previously known ranges), except for

vouchers in the collection of the ICN that erroneously cite

the municipality of Villavicencio in the Meta Department

for some locality data. For example, ICN-MHN-Rep

7531, 7544, 7546, 7646, 7713, 7855, 7856, and 7859

are all Trachemys venusta callirostris specimens listed

for the Meta Department. Presumably, these turtles were

collected within their natural range and transported to

the Estacion de Biologia Tropical Roberto Franco by

Federico Medem, where they were kept in captivity until

their deaths, and then deposited in the ICN collection

with the locality data indicating where they died rather

than reflecting where they had been collected.

Exotic species occasionally appear among the turtles

that are confiscated by environmental authorities as they

are being transported in Colombia. There is no way to

April 2022 | Volume 16 | Number 1 | e306

Turtles of Colombia: diversity, distribution, and conservation

estimate how many of these individuals are released into

natural habitats when authorities fail to recognize them

as exotics, but when confiscated turtles are correctly

identified as non-native species, Colombian authorities

usually relocate them to zoological parks or aquariums.

For example, individuals of the Venezuelan endemic

Mesoclemmys zuliae occasionally appear on lists of fauna

confiscated by Colombian authorities (A. Echeverry-

Alcendra, pers. comm.) and some eventually make their

way into zoological collections. Another example is

an individual of the Mediterranean Mauremys leprosa

(Bertolero and Busack 2017) that was confiscated in

Bogota in 2003 and relocated to the Barranquilla Zoo,

generating an entry for this species for Colombia in

the Global Register of Introduced and Invasive Species

records (Baptiste et al. 2018). This database also lists

Trachemys scripta elegans for Colombia. This slider

subspecies presumably entered Colombia as_ part

of the illegal pet trade. The Zoological Information

Management System (ZIMS 2021) documents that some

individuals of this exotic species are housed in some

zoos in the country. There are more ambiguous anecdotal

reports of Trachemys scripta elegans individuals living

freely in Cundinamarca Department, as well as in

portions of the Cauca and Magdalena river drainages

(Morales-Betancourt et al. 2012b). However, we found

no museum vouchers of 7rachemys scripta elegans or

any other exotic turtle species that were collected in

natural habitats in Colombia.

Conservation status update and summary

The book Libro Rojo de Reptiles de Colombia (Morales-

Betancourt et al. 2015a) updated Castafio-Mora (2002a),

and evaluated the conservation status of all turtle species

in Colombia using the most recent IUCN criteria for

the first time (IUCN 2012). However, three recent

changes in turtle taxonomy create the need to examine

their implications regarding the conservation status

of the species in Colombia. In 2015, Chelus fimbriata

was Classified as Least Concern (LC) based upon its

wide distribution and apparent abundance, despite the

recognition that the illegal pet trade poses a threat to

some populations (Morales-Betancourt et al. 2015a). The

splitting of this species into Orinoco (Chelus orinocensis)

and Amazonian (Chelus fimbriata) species (Vargas-

Ramirez et al. 2020) does not substantially modify

this assessment, and we recommend that both species

tentatively be considered as LC as well, at least until

their next formal reassessment. Similarly, the recognition

of Mesoclemmys warmuthi as a valid species and its

apparent co-occurrence with M. raniceps in the Amazon

department argues that both should be assigned a Data

Deficient (DD) status at present. Finally, Trachemys

callirostris (here Trachemys venusta callirostris) was

classified as Vulnerable (VU) in 2015, while Trachemys

venusta (here Trachemys medemi) was classified as DD

Amphib. Reptile Conserv.

(Morales-Betancourt et al. 2015a). The recent taxonomic

revision (Vargas-Ramirez et al. 2017) did not affect the

classification of either taxon, so Trachemys venusta

callirostris should continue to be considered as VU at the

national level and 7rachemys medemi should continue to

be classified as DD.

With this updated classification of the threat levels

faced by the Colombian turtle species, the sea turtles

exhibit the highest level of conservation concern, with

all five species categorized in one of the three threatened

categories (VU, EN, or CR; IUCN 2012). Next is the

family Podocnemididae, with four of its seven species

(54%) being classified in a threatened category. In terms

of macro-drainages, the CAR macro-drainage is the

most impacted, with 43% of its turtle species considered

threatened, while the AMA macro-drainage is at the

other extreme, with only 16% of its species considered

as threatened. Finally, in terms of the 27 departments

that possess at least four turtle species, seven (Bolivar,

Cundinamarca, La Guajira, Magdalena, San Andrés,

Providencia, and Santa Catalina, Santander, and Sucre)

exhibit the highest proportions of turtle species facing

conservation concerns (more than 60%), followed by

six departments (Antioquia, Atlantico, Boyaca, Cesar,

Choco, and Cordoba) with more than 50% of their turtle

species categorized as facing some level of threat. It is

relevant to examine the threat levels by department in

Colombia because it is usually at this local level that

conservation decisions are made and resources are

appropriated for management actions.

With respect to bordering countries, all four South

American countries that share borders with Colombia have

recently published updated turtle species checklists which

include evaluations of their conservation status at the

national level (Venezuela: Rodriguez et al. 2015; Brazil:

ICMBio 2018, Costa et al. 2022; Peru: SERFOR 2018;

Ecuador: assessment from IUCN Red List 2018, Torres-

Carvajal et al. 2019). Together, these five countries are the

most diverse in terms of turtle species in South America.

The results of our update are compared to those of these

other four species-rich neighboring countries in Table 2.

With 33 species, Colombia ranks second behind Brazil

(38) in turtle species richness. All five countries largely

concur that their sea turtle species should be considered

as threatened, but Brazil does not classify any of their

podocnemidid species as threatened, while almost all

species in this family in the remaining four countries are

classified nationally as either VU, EN, or CR (except for

two species classified as DD in Colombia).

Discussion

Although turtles are relatively large and conspicuous, as

well as ecologically, and often economically, important

(Lovich et al 2018), they are poorly represented in reptile

collections in general (~4% of all specimens; Lehn et al.

2007). Most turtle species are easily identifiable even as

April 2022 | Volume 16 | Number 1 | e306

Paez et al.

Table 2. Extant turtle species that occur in the five most species-rich South American countries and their global and national

conservation status. Cells in gray indicate species that are categorized as facing some level of threat by that country. Conservation

status at the national level as taken from: Colombia: Morales-Betancourt et al. 2015; Venezuela: Rodriguez et al. 2015; Brazil: Livro

Vermelho da Fauna Brasileira Amenagada de Extincao 2018; Peru: SERFOR 2018; Ecuador: Torres-Carvayjal et al. 2019. Global

conservation status based upon the IUCN Red List (http://www. iucnredlist.org).

Taxon Brazil Colombia Ecuador Peru Venezuela Global status

Suborder Cryptodira

Family Chelydridae

1. Chelydra acutirostris xX x NE

Family Geoemydidae

2. Rhinoclemmys annulata x x NT

3. Rhinoclemmys diademata a: aa | Se LC

4. Rhinoclemmys melanosterna x x LG

5. Rhinoclemmys nasuta x x NT

6. Rhinoclemmys punctularia x xX LC

Family Emydidae

7. Trachemys adiutrix | a7 EN

8. Trachemys dorbigni x LC

9. Trachemys medemi sp. nov. x NE

10. Trachemys venusta (T. v.

callirostris in Colombia, T. v.

chichiriviche in Venezuela)

Family Kinosternidae

11. Kinosternon dunni

12. Kinosternon leucostomum xX xX Le

13. Kinosternon scorpioides x x x x x LC

Family Testudinidae

14. Chelonoidis carbonarius x x Le

15. Chelonoidis denticulatus x xX xX VU

16. Chelonoidis niger Varies among the

subspecies

Family Cheloniidae

17. Caretta caretta

18. Chelonia mydas

19. Eretmochelys imbricata

20. Lepidochelys olivacea

Family Dermochelyidae

21. Dermochelys coriacea

Suborden Pleurodira

Family Chelidae

22. Acanthochelys x NT

macrocephala

23. Acanthochelys radiolata x NT

24. Acanthochelys spixii x NT

25. Chelus fimbriata x x x xX LC

26. Chelus orinocensis sp. nov. x x x EC

27. Mesoclemmys dahli [te

28. Mesoclemmys gibba x x x x x LC

Amphib. Reptile Conserv. 119 April 2022 | Volume 16 | Number 1 | e306

Turtles of Colombia: diversity, distribution, and conservation

Table 2 (continued). Extant turtle species that occur in the five most species-rich South American countries and their global and

national conservation status. Cells in gray indicate species that are categorized as facing some level of threat by that country.

Conservation status at the national level as taken from: Colombia: Morales-Betancourt et al. 2015; Venezuela: Rodriguez et al.

2015; Brazil: Livro Vermelho da Fauna Brasileira Amenacada de Extin¢ao 2018; Peru: SERFOR 2018; Ecuador: Torres-Carvajal et

al. 2019. Global conservation status based upon the IUCN Red List (http://www. iucnredlist.org).

Taxon Brazil Colombia

29. Mesoclemmys jurutiensis xX

sp. nov.

30. Mesoclemmys nasuta

31. Mesoclemmys perplexa

32. Mesoclemmys raniceps

33. Mesoclemmys tuberculata

34. Mesoclemmys vanderhaegei

x KK KK

35. Mesoclemmys wermuthi

36. Mesoclemmys zuliae

37. Phrynops geoffroanus

38. Phrynops hilarii

39. Phrynops tuberosus

40. Phrynops williamsi

41. Platemys platycephala

42. Ranacephala hogei

43. Rhinemys rufipes

44. Hydromedusa maximiliani

KS MM MMM XK

45. Hydromedusa tectifera

Family Podocnemididae

46. Peltocephalus dumerilianus

47. Podocnemis erythrocephala

48. Podocnemis expansa xX

49. Podocnemis lewyana

50. Podocnemis sextuberculata x

51. Podocnemis unifilis xX

52. Podocnemis vogli

Ol KK <M KK RK OX

Total number of families 8

Total number of genera 20 17

Total number of species 38 33

% Threatened species 24.3 42.4

neonates, so space limitations cannot fully explain their

scarcity in collections. Vouchering neonates would also

have a limited demographic impact on threatened turtle

populations (Heppell 1998). Therefore, we urge turtle

biologists to be more aware of the need to deposit voucher

specimens 1n museums in order to better document

the distributions of the species they are studying,

and to allow the re-examination of specimens when

taxonomic changes occur. For example, in Colombia

there is currently a need to reassess Matamata and M.

raniceps voucher specimens in collections, in view of

the recent split of Chelus fimbriatus into two species

and the resurrection of M. wermuthi as a valid species

separate from M. raniceps (Vargas-Ramirez et al.

Amphib. Reptile Conserv.

Ecuador Peru Venezuela Global status

Xx x Le

x NE

x VU

x x x LC

x NE

x 4 x ke.

x x 4 VU

x VU

x x 4 LR/cd

x VU

x 4 x VU

xX LC

8 6 8

15 13 15

21 17 22

42.8 47.0 47.8

2020; Cunha et al. 2019, 2021). Such a reassessment

would permit a re-definition of their documented

distributions in Colombia and identify possible areas

of sympatry. Re-examination of Colombian slider

turtle voucher specimens would also be desirable

now that the two Trachemys taxa in Colombia have

been shown to be valid and distinctive species,

rather than merely subspecies (Vargas-Ramirez et al.

2017), to better determine their range limits and/or

identify possible zones of contact. Having accurate

distribution information on turtle species is relevant

to conservation, given that three of the five criteria the

IUCN employs to categorize threat levels for species

consider the size of the range of the species being

April 2022 | Volume 16 | Number 1 | e306

Paez et al.

classified (IUCN 2012).

We also encourage future authors to report voucher

specimens as support for the inclusion of each species

in their inventory checklists. Erroneous, unsubstantiated

reports tend to self-perpetuate in the literature when

rigorous supporting evidence is not proffered. In this

checklist, despite our conservative approach of only

including species that were supported by museum

vouchers and scientific publications, we have shown

Colombia to house a large and diverse turtle fauna that

includes species from nine families. The evidence of

occurrence of the non-marine turtle species within the

five main hydrological drainages in Colombia was well

supported both by vouchers and literature, but many gaps

remain in terms of not finding vouchers to support the

occurrence of various turtle species at the department

level. Surprisingly, sea turtles were among the species

most poorly documented by specimens in museum

collections in terms of departmental occurrence, despite

the attention they receive from conservation NGOs.

The failure of sea turtle biologists to deposit voucher

specimens in biological collections in the past means that

now we cannot distinguish between the two explanations

for the disparities in reports of where certain species

nest (i.e., erroneous historical reports of nesting versus

extirpation of these populations during the past decades).

Photographic vouchers of species occurrences

uploaded to the Internet represent another means to

incorporate citizen science into species distribution

databases (Brown and Williams 2018), but we

made sparing use of such records here. Some online

records either include photographs that do not show

taxonomically important characters, fail to mention

whether the individual in the photograph was part of a

captive collection, or only provide “obscured” geographic

data to avoid revealing the precise locality information

to potential commercial collectors. While sites such as

iNaturalist offer opportunities to refine our information

on the distributions of Colombian turtle species, care

should be used in evaluating such evidence (Tiago et

al. 2017). Photographic vouchers are complementary,

but should never be considered as a replacement for the

scientific collecting of vouchers for deposit in curated

biological collections.

We have probably failed to locate some important

voucher specimens during our searches, 1n part because

some collections in Colombia still do not publish their

voucher data online. Our hope 1s that this publication will

stimulate others to make currently “hidden” voucher data

accessible, as well as to continue updating the voucher

information now contained in Table 1. Such efforts

will provide rigorous evidence for the occurrence of all

species in all departments in Colombia, both by reporting

additional relevant vouchers that exist but we were

unable to find, and especially by encouraging targeted

collecting efforts in the departments that now genuinely

lack vouchers. Additional collecting efforts may also

Amphib. Reptile Conserv.

add new species to our national inventory for Colombia,

such as Mesoclemmys gibba in the Vichada department,

Mesoclemmys zuliae in the Cesar and Norte de Santander

departments, and perhaps Trachemys venusta venusta or

Trachemys grayi in the Choco region along the Caribbean

or Pacific borders with Panama, respectively.

Hopefully, future scientific collecting efforts for

turtles will also routinely include the deposition of

genetic samples in museum collections along with the

preserved specimens, as genetic data can reveal “cryptic”

species (Vargas-Ramirez et al. 2020) and evolutionarily

significant units (Jensen et al. 2014; Vargas-Ramirez et

al. 2010). Genetic data also may help in identifying cases

of genetic contamination, since native turtle species in

Colombia are collected and transported as part of the

illegal national and international pet trade and for the

human consumption market. For the OR and AMA

basins, species such as C. fimbriata and C. orinocensis

are illegally harvested and transported to Leticia for

export to Peru, a country where the international turtle

trade is legal, unlike Colombia (Lasso et al. 2018).

For example, records indicate that exports of P. unifilis

from Peru to Hong Kong and China have increased up

to 190-fold in less than a decade (Sinovas et al. 2017).

Unfortunately, there are no reliable data on the magnitude

of harvests for most species, but the data that do exist

on the confiscation rates of illegally transported turtles

suggest the harvest is massive (Arroyave et al. 2014;

Lasso et al. 2018; Morales-Betancourt et al. 2012a). In

many cases, environmental authorities in Colombia do

not record information on confiscations (or voluntary

surrendering of illegal wildlife pets) in a standardized

manner, but the limited information available (Morales-

Betancourt et al. 2012a) suggests that the most widely

trafficked species are (in descending order) 7rachemys

venusta callirostris, Podocnemis unifilis, Chelonoidis

carbonarius, and Podocnemis expansa.

Environmental authorities in Colombia often fail to

and relocate for reinforcement or reintroduction in

a consistent manner (IUCN 2013). Individuals of

native turtle species are sometimes released within

the jurisdiction of the environmental authority that

confiscated the turtles, despite a lack of information

on the provenance of the individuals. These authorities

also transfer many confiscated turtles, especially those

confiscated in urban centers, to other areas for release,

again without knowing the exact provenance of the

individuals. In addition, people who buy turtles as pets

in Colombia (which is an illegal practice) or receive

them as gifts often decide later to release them in natural

habitats, again without knowing their origin. Thus, the

risks of genetic contamination and/or artificial range

expansion for native turtle species in Colombia appear

to be high. Although fast, cost-effective, and practical

genetic protocols have recently been developed to

aid in identifying the source of confiscated Matamata

April 2022 | Volume 16 | Number 1 | e306

Turtles of Colombia: diversity, distribution, and conservation

turtles in Colombia, at least to the level of the correct

river basin (Cardefiosa et al. 2021), much work remains

before similar protocols are available for the majority of

species that are subjected to illegal harvest and transport

in the country. Such efforts not only help to avoid genetic

contamination of native populations, but also reduce the

time and cost of maintaining individuals in captivity,

reducing the health and welfare risks to these turtles.

Our analyses revealed considerable geographic

heterogeneity in turtle species richness in Colombia,

and they also revealed substantial variation in the

conservation status of the turtles that occupy different

regions of the country. CAR was both the most species-

rich of the five macro-drainages and the one with the

greatest percentage of threatened turtle species. This

region of Colombia has suffered from substantial

perturbation and loss of natural habitats (Correa-Ayram

et al. 2018). It is also where the custom of consuming

turtles is culturally the strongest, especially during lent

(Morales-Betancourt et al. 2012a). In terms of taxonomy,

the most threatened turtle species in Colombia are the

sea turtles and podocnemidids (i.e., the largest species),

suggesting that harvest for human consumption 1s a more

important factor than collecting for the pet trade, as is the

case for most other large vertebrate species in the world

(Ripple et al. 2017). The national threat classifications

for turtles in Colombia are comparable to similar national

classifications for the same species in neighboring

countries, with the exception of Brazil where the threat

levels of podocnemidids are considered to be lower for

some reason. The only other divergence with respect to

the classifications of podocnemidids was Colombia’s

classifications of Peltocephalus dumerilianus and

Podocnemis sextuberculata as DD. We suspect that when

more data on these two species in Colombia become

available, they will be updated to receive a classification

of some level of threat, as in the neighboring countries

(excluding Brazil).

About 14% of Colombia’s maritime territory is

designated as protected areas (RUNAP 2018), so all sea

turtle species have ranges that contain some refuge from

harvest. The percentage of terrestrial territory in protected

areas in Colombia is slightly higher (~16%), yet it fails to

afford protection to all of the non-marine turtle species.

Forero-Medina et al. (2014) reported that only 15 of the

25 non-marine Colombian turtle species they evaluated

had records documenting their occurrence in protected

areas (PAs); however, they noted that information on

the distribution of some of these species predicted their

undocumented occurrences in these PAs. The situation

of having better documentation of species occurrences

outside of PAs may reflect the historical difficulties

associated with collecting in national parks and other

PAs in Colombia, due to both existing legislation (MADS

2015, 2016) and restricted access to these areas due to

armed conflicts (Negret et al. 2017).

It has long been recognized that conservation strategies

Amphib. Reptile Conserv.

cannot rely exclusively on the existence of nature reserves

(Western 1989; DeClerck et al. 2010; Morales-Betancourt

and Lasso 2015a). In Colombia, many protected areas

are also recognized as reserves where ethnic groups

are allowed to engage in subsistence hunting (Moreno

and Negrete 2012) and/or are located in regions where

armed conflict occurs (Davalos 2001; Clerici et al. 2020;

Liévano-Latorre et al. 2021), making the protection of

turtle species in these areas difficult. In addition, nature

reserves are not insurance against possible impacts on

turtle populations from global climate change (Ihlow et

al. 2012). Protecting the rich turtle fauna of Colombia

will require monitoring of the populations both within

(Laurance et al. 2012) and outside of reserves, and

implementing effective mitigation efforts when declines

are detected. Continued vouchering of the distributions

of Colombian turtles should be a part of this effort, as

evidence of range declines is one of the most compelling

indicators that a species is becoming threatened (IUCN

2012).

Conclusions

Here we present an updated, annotated checklist of the

turtle species of Colombia, compiled using a conservative

approach that only includes species documented by

museum vouchers and peer-reviewed scientific literature.

Colombia includes 33 turtle species belonging to nine

families. We also assessed the quality of the evidence

for the occurrence of each of these species in the five

major hydrological drainages and each department in

Colombia. Occurrence in the drainages was confirmed

by vouchers and the literature, but there are gaps in

terms of evidence for the occurrence of some species

in some departments. We evaluated the threat levels

for turtle species in Colombia based on taxonomy and

geographic regions, and urge biologists to recognize the

importance of vouchering specimens of turtle species

in biological collections. A better documentation of the

distributions of these species, and changes in their range

sizes, is essential for correctly classifying their threat

levels and for reducing the number of species that must

be designated as Data Deficient.

Acknowledgements.—John Carr helped improve this

manuscript in many ways. Andrés Camilo Montes

shared his opinions about the validity of the distribution

data for some species. We also would like to thank all

our colleagues who have helped make the databases we

consulted available on the Internet.

Literature Cited

Acufia- Vargas JC. 2016. Anfibios y reptiles asociados a

cinco coberturas de la tierra, municipio de Dibulla, La

Guajira, Colombia. Acta Zoolégica Mexicana 32(2):

133-146.

April 2022 | Volume 16 | Number 1 | e306

Paez et al.

Adames-Jiménez TA, Rivera-Robles DL, Arango-

Lozano J, Botero-Botero A. 2018. New records of the

turtle ZTrachemys venusta callirostris in the Central

Andes of Colombia. The Herpetological Bulletin

144(3): 28-29.

Alfaro JP, Acosta-G AE, Vejarano M. 2011. Reptiles del

Casanare. Pp. 153-167 In: Biodiversidad de Casanare:

Ecosistemas _Estratégicos del Departamento.

Gobernacion del Casanare, WWF-Colombia, Bogota,

Colombia. 286 p.

Alvarez-Rodriguez RA, Ramirez-Gallego C, Restrepo-

Garzon N, Ruiz-Alvarez T, Barrientos-Mufioz KG.

2021. Conservacion de la tortuga carey, Eretmochelys

imbricata, en Rincon del Mar, Colombia. Boletin de

Investigaciones Marinas y Costeras 50(2): 53-72.

Amorocho DF, Rubio TH, Diaz RW. 1992. Observaciones

sobre el estado actual de las tortugas marinas en el

Pacifico Colombiano. Pp. 155-179 In: Contribucion al

Conocimiento de las Tortugas Marinas de Colombia.

Editors, Rodriguez-Machado JV, Sanchez-Paez, H.

Inderena, Bogota, Colombia. 190 p.

Amorocho DF, Reina RD. 2008. Intake passage time,

digesta composition, and digestibility in east Pacific

Green Turtles (Chelonia mydas agassizii) at Gorgona

National Park, Colombian Pacific. Journal of

Experimental Marine Biology and Ecology 360(2):

117-124.

Angarita-Sierra T. 2014. Diagnosis del estado de

conservacion del ensamble de anfibios y reptiles

presentes en los ecosistemas de sabanas inundables de

la cuenca del rio Pauto, Casanare, Colombia. Revista

de la Academia Colombiana de Ciencias Exactas,

Fisicas y Naturales 38(146): 53-78.

Angarita-Sierra T, Ospina-Sarria J, Anganoy-Criollo M,

Pedroza-Banda R, Lynch JD. 2013. Guia de Campo de

los Anfibios y Reptiles del Departamento de Casanare

(Colombia). Serie Biodiversidad para la Sociedad

Numero 2. Universidad Nacional de Colombia, Sede

Orinoquia, Arauca, Arauca Department, Colombia.

117 p.

Arango-Lozano J, Young- Valencia K, Giraldo A, Botero-

Botero A. 2017. Estructura etaria y morfometria

poblacional de la tortuga pimpano (Chelydra

acutirostris, Peters 1862) (Chelydridae) en tres

afluentes del rio La Vieja, Departamento de Quindio,

Colombia. Boletin Cientifica del Museo de Historia

Natural 21(1): 122-137.

Arango-Lozano J, Patifio D, Benitez L, Botero-Botero

A. 2018. New records of Kinosternon lecuostomum

postinguinale (Duméril y Bibron, 1851) from the

Central Cordillera of Colombia. Revista Colombiana

de Ciencias Animales 10(1): 82-85.

Armesto LO, Gutiérrez DR, Pacheco RD, Gallardo AO.

2011. Reptiles from the municipality of Cucuta (Norte

de Santander, Colombia). Boletin Cientifico. Centro de

Museos. Museo de Historia Natural 15(2): 157-168.

Arroyave FJ, Romero-Goyeneche OY, Bonilla-Gomez

Amphib. Reptile Conserv.

MA, Hurtado-Heredia RG. 2014. Trafico ilegal de

tortugas continentales (Testudinata) en Colombia:

una aproximacion desde el andalisis de redes. Acta

Biologica Colombiana 19(3): 381-392.

Baptiste MP, Garcia LM, Acevedo-Charry O, Acosta

A, Alarcon J, Arévalo E, Avella GC, Blanco A,

Botero JE, Pagad S, et al. 2018. Global Register of

Introduced and Invasive Species — Colombia. Version

1.5. Available: http://gbif.org/168568e7-eb5f-4ef6-

8c59-f73ceaf57e91 [Accessed: 3 January 2021].

Barreto L. 2016. Conservacion de Tortugas Marinas

en Colombia. Editor, Barreto L. Fundacion

Conservacion Ambiente Colombia, Bogota,

Colombia. Available: —https://archive.org/details/

Barreto20106ConservacionTortugasMarinasEn

ColombiaVoll [Accessed: 3 January 2021 |

Barrientos-Mufioz KG, Ramirez-Gallego C, Rivas L.

2013. First report of nesting of the Black Sea Turtle

(Chelonia mydas) on the north Pacific coast of

Colombia. Marine Turtle Newsletter 138: 19-21.

Barrientos-Mufioz KG, Ramirez-Gallego C, Paez VP.

2014. Nesting ecology of the Olive Ridley Sea Turtle

(Lepidochelys olivacea) (Cheloniidae) at El Valle

Beach, northern Pacific, Colombia. Acta Bioldgica

Colombiana 19(3): 437-445.

Barrientos-Mufioz KG, Ramirez-Gallego C, Paez VP.

2015a. Eretmochelys imbricata (Linnaeus, 1766).

Pp. 127-131 In: Libro Rojo de Reptiles de Colombia

(2015). Editors, Morales-Betancourt MA, Lasso

CA, Paez VP, Bock BC. Instituto de Investigacion

de Recursos Biologicos Alexander von Humboldt,

Bogota, Colombia. 258 p.

Barrientos-Mufioz KG, Ramirez-Gallego C, Paez VP.

2015b. Lepidochelys olivacea (Eschscholtz, 1829).

Pp. 161-165 In: Libro Rojo de Reptiles de Colombia

(2015). Editors, Morales-Betancourt MA, Lasso

CA, Paez VP, Bock BC. Instituto de Investigacion

de Recursos Biologicos Alexander von Humboldt,

Bogota, Colombia. 258 p.

Barrientos-Mufioz KG, Valencia M, Hinojosa CA,

Valencia-Gonzalez S, Diazgranados MC, Sevilla EL,

Ramirez-Gallego C. 2020. Primer registro de una

hembra de tortuga carey, Eretmochelys imbricata, en

el Mosaico de Conservacion Bahia Malaga, Pacifico

colombiano. Boletin de Investigaciones Marinas y

Costeras 49(Suplemento Especial): 289-294.

Barrientos-Mufioz KG, Heidemeyer M, Ortega-Gordillo

LF, Valencia-Diaz M, Hinojosa-Romero CA, Valencia-

Gonzalez S, Montafio HJ, Diazgranados MC, Caldas

JP, Ramirez-Gallego C. 2022. Tortugas marinas en

el mosaico de conservacion Bahia Malaga, Pacifico

colombiano. Boletin de Investigaciones Marinas y

Costeras. |In Press].

Bernhard R, de la Ossa-Velasquez J, Vogt RC, Morales-

Betancourt MA, Lasso CA. 2012. Podocnemis

erythrocephala (Spix 1824). Pp. 360-366 In:

V. Biologia y Conservacién de las Tortugas

April 2022 | Volume 16 | Number 1 | e306

Turtles of Colombia: diversity, distribution, and conservation

Continentales de Colombia. Serie Editorial Recursos

Hidrobiologicos y Pesqueros Continentales de

Colombia. Editors, Paez VP, Morales-Betancourt MA,

Lasso CA, Castafio-Mora OV, Bock BC. Instituto de

Investigacion de Recursos Biologicos Alexander von

Humboldt, Bogota, Colombia. 523 p.

Bertolero A, Busack SD. 2017. Mauremys

leprosa (Schweigger, 1812), Mediterranean Stripe-

necked Terrapin. Chelonian Research Monographs 5:

102.1-102.19.

Berry JF, Iverson JB, Forero-Medina G. 2012.

Kinosternon scorpioides (Linnaeus 1766). Pp. 340-

348 In: V. Biologia y Conservacion de las Tortugas

Continentales de Colombia. Serie Editorial Recursos

Hidrobiologicos y Pesqueros Continentales de

Colombia. Editors, Paez VP, Morales-Betancourt MA,

Lasso CA, Castafio-Mora OV, Bock BC. Instituto de

Investigacion de Recursos Biologicos Alexander von

Humboldt, Bogota, Colombia. 523 p.

Blanco-Torres A, Baez L, Patifio-Flores E, Renjifo-

Rey JM. 2013. Herpetofauna del valle medio del

rio Rancheria, La Guajira, Colombia. Revista de

Biodiversidad Neotropical 3(2): 113-122.

Blanco-Torres A, Duran-Prieto C, Acosta-Galvis AR.

2019. Herpetofauna. Pp. 163-187 In: Biodiversidad

en el Departamento de Arauca. Editors, Trujillo F,

Anzola F. Gobernacion de Arauca, Fundacion Omacha

y Fundacion Ecollano, Bogota, Colombia. 384 p.

Bock BC, Paez VP, Daza JM. 2012a. Trachemys

callirostris (Gray, 1856). Pp. 283-291 In: V. Biologia

y Conservacion de las Tortugas Continentales de

Colombia. Serie Editorial Recursos Hidrobiologicos

y Pesqueros Continentales de Colombia. Editors, Paez

VP, Morales-Betancourt MA, Lasso CA, Castafio-

Mora OV, Bock BC. Instituto de Investigacion de

Recursos Bioldgicos Alexander von Humboldt,

Bogota, Colombia. 523 p.

Bock BC, Paez VP, Castafio-Mora OV. 2012b. Trachemys

venusta (Gray, 1856). Pp. 292-297 In: V. Biologia

y Conservacion de las Tortugas Continentales de

Colombia. Serie Editorial Recursos Hidrobiologicos

y Pesqueros Continentales de Colombia. Editors, Paez

VP, Morales-Betancourt MA, Lasso CA, Castafio-

Mora OV, Bock BC. Instituto de Investigacion de

Recursos Bioldgicos Alexander von Humboldt,

Bogota, Colombia. 523 p.

Bock BC, Paez VP, Cortes-Duque J. 2015. Hicotea.

Trachemys callirostris (Gray, 1856). Pp. 166-171 In:

Libro Rojo de Reptiles de Colombia (2015). Editors,

Morales-Betancourt MA, Lasso CA, Paez VP, Bock

BC. Instituto de Investigacion de Recursos Bioldgicos

Alexander von Humboldt, Bogota, Colombia. 258 p.

Bock BC, Paez VP. 2017. An analysis of the history of the

study of freshwater turtles and tortoises of Colombia.

Acta Biologica Colombiana 22(1): 67—76.

Bolton AB, Martins HR. 1990. Kemp’s Ridley captured

in the Azores. Marine Turtle Newsletter 48: 23.

Amphib. Reptile Conserv.

Borrero WJ, Patifio EA, Guerra ML, Gourtyu WA. 2013.

New evidence of nesting Dermochelys coriacea

(Tortuga Achepo) at Iporoimao-Utareo beaches,

Guajira, Colombia. Marine Turtle Newsletter 137:

13-14.

Bour R, Pauler I. 1987. Indentité de Phrynops

vanderhaegei Bour, 1973 et des especes affines.

Mésogée 47: 3-23.

Brown ED, Williams BK. 2018. The potential for citizen

science to produce reliable and useful information in

ecology. Conservation Biology 33(3): 561-569.

Caceres-Martinez CH, Acevedo-Rincén AA, Sierra-

Leal JA, Gonzalez-Maya JF. 2017. Kinosternon

scorpioides scorpioides (Testudines: Kinosternidae):

nuevo reporte en el nororiente de Colombia. Acta

Biologica Colombiana 22(2): 242-245.

Camacho-Mosquera L, Amorocho DF, Mejia-Ladino

LM, Palacio-Mejia JD, Rond6on-Gonzalez F. 2008.

Caracterizacion genética de la colonia reproductiva

de la tortuga marina golfina, Lepidochelys olivacea,

en el Parque Nacional Natural Gorgona (Pacifico

colombiano) a partir de secuencias de ADN

mitocondrial. Boletin de Investigaciones Marinas y

Costeras 37(1): 77-92.

Cafias-Uribe M, Payan LF, Amorocho DF, Paez VP.

2020. Tasas de crecimiento somatico de tortuga carey,

Eretmochelys imbricata, en el Parque Nacional Natural

Gorgona, Colombia, entre 2004 y 2018. Boletin de

Investigaciones Marinas y Costeras 49(Suplemento

Especial): 13-30.

Cardefiosa D, Chapman DD, Robles YL, Ussa DA,

Caballero S. 2021. Rapid species and river-of-origin

determination for Matamata turtles (Che/us sp.) using

real-time PCR: facilitating rapid return of trafficked

specimens back to the wild. Aquatic Conservation:

Marine and Freshwater Ecosystems 31(9): 1-8.

Carr JL, Giraldo A, Garcés-Restrepo M. 2012.

Rhinoclemmys nasuta (Boulenger, 1902). Pp. 315—

322 In: V. Biologia y Conservacion de las Tortugas

Continentales de Colombia. Serie Editorial Recursos

Hidrobiolégicos y Pesqueros Continentales de

Colombia. Editors, Paez VP, Morales-Betancourt MA,

Lasso CA, Castafio-Mora OV, Bock BC. Instituto de

Investigacion de Recursos Biologicos Alexander von

Humboldt, Bogota, Colombia. 523 p.

Carvajal-Cogollo JE. 2019. Capitulo II. Taxonomia,

sistematica, y zoogeografia de reptiles. Pp. 97-161

In: Biologia de los Anfibios y Reptiles en el Bosque

Seco Tropical del Norte de Colombia. Editors, Vargas-

Salinas F, Mufioz-Avila JA, Morales-Puentes ME.

Editorial Universidad Pedagogica y Tecnologica de

Colombia, Tunja, Colombia. 484 p.

Carvalho VT. 2016. Revisaéo taxondmica e sistematica

filogenética de Phrynops geoffroanus (Schweigger,

1812) (Testudines: Chelidae). Ph.D. Dissertation,

Universidad Federal do Amazonas, Manaus, Brazil.

124 p.

April 2022 | Volume 16 | Number 1 | e306

Paez et al.

Carvalho VT, Martinez JG, Hernandez-Rangel SM,

Astolfi-Filho S, Vogt RC, Farias IP, Hrbek T. 2016.

Giving IDs to turtles: SNP markers for assignment of

individuals to lineages of the geographically structured

Phyrnops geoffroanus’ (Chelidae: _Testudines).

Conservation Genetic Resources 9: 157-163.

Castafio-Mora OV. (Editor). 2002a. Libro Rojo de Reptiles

de Colombia. Libros Rojos de Especies Amenazadas

de Colombia. Instituto de Ciencias Naturales-

Universidad Nacional de Colombia, Ministerio

del Medio Ambiente, Conservacion Internacional-

Colombia, Bogota, Colombia. 160 p.

Castafio-Mora OV. 2002b. Chelydra _ serpentina

acutirostris. Pp. 123 In: Libro Rojo de Reptiles de

Colombia. Libros Rojos de Especies Amenazadas

de Colombia. Editor, Castafio-Mora OV. Instituto de

Ciencias Naturales-Universidad Nacional de Colombia,

Ministerio del Medio Ambiente, Conservacion Inter-

nacional-Colombia, Bogota, Colombia. 160 p.

Castafio-Mora OV. 2002c. Podocnemis erythrocephala.

Pp. 109-111 In: Libro Rojo de Reptiles de Colombia.

Libros Rojos de Especies Amenazadas de Colombia.

Editor, Castafio-Mora OV. Instituto de Culencias

Naturales-Universidad Nacional de Colombia,

Ministerio de Medio Ambiente, Conservacion

Internacional-Colombia, Bogota, Colombia. 160 p.

Castafio-Mora OV, Lugo-Rugeles M. 1981. Estudio

comparativo de dos especies de morrocoy: Geochelone

carbonaria y Geochelone denticulata y aspectos

comparables de su morfologia externa. Cespedesia

10(37-38): 55-122.

Castafio-Mora OV, Medem F. 2002a. Podocnemis

lewyana. Pp. 92-94 In: Libro Rojo de Reptiles de

Colombia. Libros Rojos de Especies Amenazadas

de Colombia. Editor, Castafio-Mora OV. Instituto

de Ciencias Naturales-Universidad Nacional

de Colombia, Ministerio de Medio Ambiente,

Conservacion __Internacional-Colombia, Bogota,

Colombia. 160 p.

Castafio-Mora OV, Medem F. 2002b: Trachemys scripta

callirostris. Pp. 118-119 In: Libro Rojo de Reptiles

de Colombia. Libros Rojos de Especies Amenazadas

de Colombia. Editor, Castafio-Mora OV. Instituto

de Ciencias Naturales-Universidad Nacional

de Colombia, Ministerio de Medio Ambiente,

Conservacion _Internacional-Colombia, Bogota,

Colombia. 160 p.

Castafio-Mora OV, Medem F. 2002c. Batrachemys dahii.

Pp. 77-79 In: Libro Rojo de Reptiles de Colombia.

Libros Rojos de Especies Amenazadas de Colombia.

Editor, Castafio-Mora OV. Instituto de Culencias

Naturales-Universidad Nacional de Colombia,

Ministerio de Medio Ambiente, Conservacion

Internacional-Colombia, Bogota, Colombia. 160 p.

Castafio-Mora OV, Medem F. 2002d. Batrachemys

raniceps. Pp. 130 In: Libro Rojo de Reptiles de

Colombia. Libros Rojos de Especies Amenazadas

Amphib. Reptile Conserv.

de Colombia. Editor, Castafio-Mora OV. Instituto

de Ciencias Naturales-Universidad Nacional

de Colombia, Ministerio de Medio Ambiente,

Conservacion _Internacional-Colombia, Bogota,

Colombia 160 p.

Castafio-Mora OV, Medem F. 2002e. Rhinemys rufipes.

Pp. 98-99 In: Libro Rojo de Reptiles de Colombia.

Libros Rojos de Especies Amenazadas de Colombia.

Editor, Castafio-Mora OV. Instituto de Culencias

Naturales-Universidad Nacional de Colombia,

Ministerio de Medio Ambiente, Conservacion

Internacional-Colombia, Bogota, Colombia. 160 p.

Castafio-Mora OV, Medem F. 2002f. Podocnemis expa-

nsa. Pp. 61-64 In: Libro Rojo de Reptiles de Colombia.

Libros Rojos de Especies Amenazadas de Colombia.

Editor, Castafio-Mora OV. Instituto de Culencias

Naturales-Universidad Nacional de Colombia,

Ministerio de Medio Ambiente, Conservacion

Internacional-Colombia, Bogota, Colombia. 160 p.

Castafio-Mora OV, Medem F. 2002g. Podocnemis unifilis.

Pp. 65-67 In: Libro Rojo de Reptiles de Colombia.

Libros Rojos de Especies Amenazadas de Colombia.

Editor, Castafio-Mora OV. Instituto de Culencias

Naturales-Universidad Nacional de Colombia,

Ministerio de Medio Ambiente, Conservacion

Internacional-Colombia, Bogota, Colombia. 160 p.

Castafio-Mora OV, Medem F. 2002h. Podocnemis vogli.

Pp. 121 In: Libro Rojo de Reptiles de Colombia. Libros

Rojos de Especies Amenazadas de Colombia. Editor,

Castafio-Mora OV. Instituto de Ciencias Naturales-

Universidad Nacional de Colombia, Ministerio

de Medio Ambiente, Conservacion Internacional-

Colombia, Bogota, Colombia. 160 p.

Castafio-Mora OV, Galvis-Pefiuela PA, Molano JG. 2003.

Reproductive ecology of Podocnemis erythrocephala

(Testudines: Podocnemididae) in the Lower Inirida

River. Chelonian Conservation and Biology 4(3):

664-670.

Castafio-Mora OV, Cardenas G, Hernandez EJ, Castro-

Herrera F. 2004. Reptiles en el Choco biogeografico.

Pp. 599-631 In: El Choco Biogeografico/Costa

Pacifica: Colombia. Diversidad Bidtica IV. Editor,

Rangel-Churio JO. Universidad Nacional de

Colombia, Bogota, Colombia. 866 p.

Castafio-Mora OV, Cardenas-Arévalo G, Medina-

Rangel GF, Carvayal-Cogollo JE, Forero-Medina GA,

Gallego-Garcia N, Ulloa-Delgado G, Rojas-Murcia

LE, Gaitan-Guerron J. 2015. Chelonoidis carbonarius

Spix 1824. Pp. 172-175 In: Libro Rojo de Reptiles

de Colombia (2015). Editors, Morales-Betancourt

MA, Lasso CA, Paez VP, Bock BC. Instituto de

Investigacion de Recursos Biologicos Alexander von

Humboldt, Bogota, Colombia. 258 p.

Castro-Herrera F, Vargas-Salinas F. 2008. Anfibios

y reptiles en el departamento del Valle del Cauca,

Colombia. Biota Colombiana 9(2): 251-277.

Ceballos-Fonseca CP. 2000. Tortugas (Testudinata)

April 2022 | Volume 16 | Number 1 | e306

Turtles of Colombia: diversity, distribution, and conservation

marinas y continentales de Colombia. Biota

Colombiana 1(2): 187-194.

Ceballos-Fonseca CP. 2004. Distribucién de playas de

anidacion y areas de alimentacion de tortugas marinas

y sus amenazas en el Caribe colombiano. Boletin de

Investigaciones Marinas y Costeras 33(1): 79-99.

Ceballos-Fonseca CP, Hernandez O, Morales-Betancourt

MA, Trujillo F. 2012a. Podocnemis expansa

(Schweigger, 1812). Pp. 367-374 In: K Biologia

y Conservacion de las Tortugas Continentales de

Colombia. Serie Editorial Recursos Hidrobiologicos

y Pesqueros Continentales de Colombia. Editors, Paez

VP, Morales-Betancourt MA, Lasso CA, Castafio-

Mora OV, Bock BC. Instituto de Investigacion de

Recursos Bioldgicos Alexander von Humboldt,

Bogota, Colombia. 523 p.

Ceballos-Fonseca CP, Iverson JB, Bernhard R. 2012b.

Podocnemis sextuberculata (Cornalia, 1849). Pp.

382-386 In: V. Biologia y Conservacion de las

Tortugas Continentales de Colombia. Serie Editorial

Recursos Hidrobiologicos y Pesqueros Continentales

de Colombia. Editors, Paez VP, Morales-Betancourt

MA, Lasso CA, Castafio-Mora OV, Bock BC. Instituto

de Investigacién de Recursos Biologicos Alexander

von Humboldt, Bogota, Colombia. 523 p.

Ceballos-Fonseca CP, Brand WA. 2014. Morphology and

conservation of the Mesoamerican Slider (7rachemys

venusta, Emydidae) from the Atrato river basin,

Colombia. Acta Biolégica Colombiana 19(3): 483-

498.

Clerici N, Armenteras D, Karieva P, Botero R, Ramirez-

Delgado JP, Forero-Medina G, Ochoa J, Pedraza C,

Schneider L, Biggs D, et al. 2020. Deforestation in

Colombian protected areas increases during post-

conflict periods. Scientific Reports 10: 4971.

Correa-Ayram CA, Diaz-Timoté J, Etter Rothlisberger

A, Ramirez W, Corzo G. 2018. El cambio en la

huella espacial humana como herramienta para la

toma de decisiones en la gestion del territorio. In:

Biodiversidad 2018. Estado y Tendencias de la

Biodiversidad Continental de Colombia. Editors,

Moreno OA, Andrade GI, Goméz MF. Instituto de

Investigaciones de Recursos Biolégicos Alexander

von Humboldt, Bogota, Colombia. Unpaginated.

Costa HC, Guedes TB, Bérnils RS. 2022. Lista de

répteis do Brasil: padrées e tendéncias. Herpetologia

Brasileira 10(3): 110-279.

Covelo P, Nicolau P, Lopez A. 2016. Four new records

of stranded Kemp’s Ridley Turtle Lepidochelys kempi

in the NW Iberian Peninsula. Marine Biodiversity

Records 9(1): 80.

Cunha FAG, Fernandes T, Franco J, Vogt RC. 2019.

Reproductive biology and hatchling morphology

of the Amazon Toad-headed Turtle (Mesoclemmys

raniceps) (Testudines: Chelidae), with notes on species

morphology and taxonomy of the Mesoclemmys

group. Chelonian Conservation and Biology 18(2):

Amphib. Reptile Conserv.

195-209.

Cunha FAG, Fagundes CK, Brito ES, Vogt RC, Maffei

F, Pezzuti J, Félix-Silva D, Rojas-Runjaic FJM, Lasso

CA, Morales-Betancourt MA, et al. 2021. Distribution

of Chelus fimbriata and Chelus orinocensis

(Testudines: Chelidae). Chelonian Conservation and

Biology 20(1): 109-115.

Davalos LM. 2001. The San Lucas Mountain range in

Colombia: How much conservation is owed to the

violence? Biodiversity and Conservation 10: 69-78.

de la Ossa-Velasquez J, Bernhard R, de la Ossa-Lacayo

A. 2012a. Platemys platycephala (Schneider, 1792).

Pp. 261-265 In: V. Biologia y Conservacion de las

Tortugas Continentales de Colombia. Serie Editorial

Recursos Hidrobiologicos y Pesqueros Continentales

de Colombia. Editors, Paez VP, Morales-Betancourt

MA, Lasso CA, Castafio-Mora OV, Bock BC. Instituto

de Investigacién de Recursos Biologicos Alexander

von Humboldt, Bogota, Colombia. 523 p.

de la Ossa-Velasquez J, Vogt RC, de la Ossa-Lacayo

A, Lasso CA. 2012b. Peltocephalus dumerilianus

(Schweigger, 1812). Pp. 353-359 In: K Biologia

y Conservacion de las Tortugas Continentales de

Colombia. Serie Editorial Recursos Hidrobiologicos

y Pesqueros Continentales de Colombia. Editors, Paez

VP, Morales-Betancourt MA, Lasso CA, Castafio-

Mora OV, Bock BC. Instituto de Investigacion de

Recursos Bioldgicos Alexander von Humboldt,

Bogota, Colombia. 523 p.

DeClerck FAJ, Chazdon R, Holl KD, Milder JC, Finegan

B, Martinez-Salinas A, Imbach P, Canet L, Ramos Z.

2010. Biodiversity conservation in human-modified

landscapes of Mesoamerica: past, present, and future.

Biological Conservation 143(10): 2,301—2,313.

del Rio J, Zapata-Ciro J, Paez VP. 2018. Podocnemis

expansa (Schweigger, 1812). Catalogo de Anfibios y

Reptiles de Colombia 4(1): 52-59.

Duellman WE. 1978. The biology of an equatorial

herpetofauna in Amazonian Ecuador. Miscellaneous

Publications of the University of Kansas Museum of

Natural History 65: 1-352.

Duefiez-Gomez F, Mufioz-Guerrero J, Ramirez-Pinilla

MP. 2004. Herpetofauna del Corregimiento Botillero

(El Banco, Magdalena) en la Depresi6n Momposina

de la region Caribe Colombiana. Actualidades

Biologicas 26(81): 161-170.

Echeverry-Alcendra AM, Guzman-M A, Stevenson

P, Cortés-Duque J. 2012. Chelenoidis denticulata

(Linnaeus, 1766). Pp. 412-418 In: V Biologia y

Conservacion de las Tortugas Continentales de

Colombia. Serie Editorial Recursos Hidrobiologicos

y Pesqueros Continentales de Colombia. Editors, Paez

VP, Morales-Betancourt MA, Lasso CA, Castafio-

Mora OV, Bock BC. Instituto de Investigacion de

Recursos Bioldgicos Alexander von Humboldt,

Bogota, Colombia. 523 p.

Echeverry-Alcendra A. 2019. Chelonoidis carbonarius

April 2022 | Volume 16 | Number 1 | e306

Paez et al.

(Spix, 1824). Catalogo de Anfibios y Reptiles de

Colombia 5(1): 13-29.

Echeverry-Garcia LP, Carr JL, Garcés-Restrepo MF,

Galvis-Rizo CA, Giraldo A. 2012a. Rhinoclemmys

melanosterna (Gray, 1861). Pp. 308-314 In:

V. Biologia y Conservacién de las Tortugas

Continentales de Colombia. Serie Editorial Recursos

Hidrobiol6gicos y Pesqueros Continentales de

Colombia. Editors, Paez VP, Morales-Betancourt MA,

Lasso CA, Castafio-Mora OV, Bock BC. Instituto de

Investigacion de Recursos Biologicos Alexander von

Humboldt, Bogota, Colombia. 523 p.

Eckert K, Eckert A. 2019. An Atlas of Sea Turtle Nesting

Habitat for the Wider Caribbean Region. WIDECAST

Technical Report Number 19. Wider Caribbean

Sea Turtle Conservation Network, Beaufort, North

Carolina, USA. 232 p.

Escalona T, Conway-Gomez K, Morales-Betancourt

MA, Arbelaez F, Antelo R. 2012. Podocnemis unifilis

(Troschel, 1848). Pp. 387-398 In: V. Biologia y

Conservacion de las Tortugas Continentales de

Colombia. Serie Editorial Recursos Hidrobiologicos

y Pesqueros Continentales de Colombia. Editors, Paez

VP, Morales-Betancourt MA, Lasso CA, Castafio-

Mora OV, Bock BC. Instituto de Investigacion de

Recursos Biolégicos Alexander von Humboldt,

Bogota, Colombia. 523 p.

ESRI. 2014. Environmental Systems Research Institute

homepage. Available: http://esri.com/en-us/home

[Accessed: 3 January 2021].

Ferrara CR, Fagundes CK, Morcatty TQ, Vogt RC.

2017. Quelénios Amazénicos. Guia de Identificacdo e

Distribuigdo. Wildlife Conservation Society, Manaus,

Brazil. 180 p.

Florez-Jaramillo LA, Barona-Cortés E. 2016. Diversity

of reptiles associated with three contrasting areas in a

tropical dry forest (La Dorada and Victoria, Caldas).

Revista de Ciencias 20(2): 109-123.

Forero-Medina G, Castafio-Mora OV. 2011. Kinosternon

scorpioides albogulare (Dumeéril and Bocourt, 1870),

White-throated Mud Turtle, Swanka Turtle. Chelonian

Research Monographs 5. 064.1—064.5.

Forero-Medina G, Castafio-Mora OV, Cardenas-Arévalo

G, Medina-Rangel GF. 2012. Mesoclemmys dahli

(Zangerl y Medem, 1958). Pp. 247—253 In: V. Biologia

y Conservacion de las Tortugas Continentales de

Colombia. Serie Editorial Recursos Hidrobiologicos

y Pesqueros Continentales de Colombia. Editors, Paez

VP, Morales-Betancourt MA, Lasso CA, Castafio-

Mora OV, Bock BC. Instituto de Investigacion de

Recursos Biolégicos Alexander von Humboldt,

Bogota, Colombia. 523 p.

Forero—Medina G, Yusti-Mufioz AP, Castafio—Mora

OV. 2014. Distribucién geografica de las tortugas

continentales de Colombia y su representaciOn en

areas protegidas. Acta Biolégica Colombiana 19(3):

415-426.

Amphib. Reptile Conserv.

Forero-Medina G, Iverson JB, Carr JL, Castafio-Mora

OV, Galvis-Rizo CA, Renteria-Moreno LE. 2015a.

Tortuga cabeza de trozo. Kinosternon dunni (Schmidt,

1947). Pp. 180-182 In: Libro Rojo de Reptiles de

Colombia (2015). Editors, Morales-Betancourt

MA, Lasso CA, Paez VP, Bock BC. Instituto de

Investigacion de Recursos Biologicos Alexander von

Humboldt, Bogota, Colombia. 258 p.

Forero-Medina G, Castafio-Mora OV, Pacheco A,

Lasso CA. 2015b. Swanka. Kinosternon scorpioides

albogulare (Duméril y Bocourt, 1870). Pp. 176-179

In: Libro Rojo de Reptiles de Colombia (2015). Editors,

Morales-Betancourt MA, Lasso CA, Paez VP, Bock

BC. Instituto de Investigacion de Recursos Biolégicos

Alexander von Humboldt, Bogota, Colombia. 258 p.

Forero-Medina G, Castafio-Mora OV, Cardenas-Arévalo

G, Medina-Rangel GF, de la Ossa- Velasquez J, Vargas-

Ramirez M, Gallego-Garcia N. 2015c. Carranchina.

Mesoclemmys dahli (Zangerl y Medem, 1958). Pp.

142-145 In: Libro Rojo de Reptiles de Colombia

(2015). Editors, Morales-Betancourt MA, Lasso

CA, Paez VP, Bock BC. Instituto de Investigacion

de Recursos Biologicos Alexander von Humboldt,

Bogota, Colombia. 258 p.

Forero-Medina G, Paez VP, Garcés-Restrepo MF, Carr

JL, Giraldo A, Vargas-Ramirez M. 2016. Research and

conservation priorities for tortoises and freshwater

turtles of Colombia. Tropical Conservation Science

9(4): 1-14.

Franco-Espinosa C, Hernandez-Fernandez J. 2017.

Analisis de haplotipos de la tortuga cabezona

Caretta caretta (Testudines: Cheloniidae) en dos

playas del Caribe colombiano. Revista Lasallista de

Investigacion 14(2): 121-131.

Friol NR. 2014. Filogenia e evolucao das espécies do

género Phrynops (Testudines: Chelidae). M.S. Thesis,

Universidade de Sao Paolo, Sao Paolo, Brazil. 101 p.

Fritz U, Havass P. 2007. Checklist of chelonians of the

world. Vertebrate Zoology 57(2): 149-368.

Fritz U, Stuckas H, Vargas-Ramirez M, Hundsdorfer

AK, Maran J, Packert M. 2012. Molecular phylogeny

of Central and South American slider turtles:

implications for biogeography and_ systematics

(Testudines: Emydidae: Trachemys). Journal of

Zoological Systematics and Evolutionary Research

50(2): 125-136.

Gallego-Garcia N, Cardenas-Arévalo G, Castafio-Mora

OV. 2012. Chelonoidis carbonaria (Spix, 1824). Pp.

406-411 In: Libro Rojo de Reptiles de Colombia

(2015). Editors, Morales-Betancourt MA, Lasso

CA, Paez VP, Bock BC. Instituto de Investigacion

de Recursos Biologicos Alexander von Humboldt,

Bogota, Colombia. 258 p.

Gaos AR, Abreu-Grobois FA, Alfaro-Shigueto J,

Amorocho D, Arauz R, Baquero A, Brisefio R, Chacon

D, Duefias C, Zarate P, et al. 2010. Signs of hope in

the eastern Pacific: international collaboration reveals

April 2022 | Volume 16 | Number 1 | e306

Turtles of Colombia: diversity, distribution, and conservation

encouraging status for a severely depleted population

of Hawksbill Turtles, Eretmochelys imbricata. Oryx

44(4): 595-601.

Garcés-Restrepo MF, Giraldo A, Carr JL. 2013.

Population ecology and morphometric variation of the

Chocoan River Turtle (Rhinoclemmys nasuta) from

two localities on the Colombian Pacific coast. Boletin

Cientifico. Centro de Museos. Museo de Historia

Natural 17(2): 160-171.

Garcés-Restrepo MF, Carr JL, Giraldo A. 2019. Long-

term variation in survival of a neotropical freshwater

turtle: habitat and climatic influences. Diversity 11(6):

1-12.

Garcia N. 2005. Biologia reproductiva y conservacion

de las tortugas charapa Podocnemis expansa, cupiso

Podocnemis sextuberculata y taricaya Podocnemis

unifilis en las playas aledafias al municipio de Puerto

Narifio (Amazonas). Undergraduate Thesis, Pontificia

Universidad Javeriana, Bogota, Colombia. 209 p.

Gaston KJ. 2003. The Structure and Dynamics of

Geographic Ranges. Oxford University Press, Oxford,

United Kingdom. 280 p.

GBIF.org. 2021. Global Biodiversity Information Facility

homepage. Available: https//www.gbif.org [Accessed:

3 January 2021].

Giraldo A, Carr JL, Garcés-Restrepo MF. 2012a.

Rhinoclemmys annulata (Gray, 1860). Pp. 300-304

In: V. Biologia y Conservacion de las Tortugas

Continentales de Colombia. Serie Editorial Recursos

Hidrobiologicos y Pesqueros Continentales de

Colombia. Editors, Paez VP, Morales-Betancourt MA,

Lasso CA, Castafio-Mora OV, Bock BC. Instituto de

Investigacion de Recursos Biologicos Alexander von

Humboldt, Bogota, Colombia. 523 p.

Giraldo A, Garcés-Restrepo MF, Carr JL. 2012b. Tamafio

y estructura poblacional de la tortuga sabaletera

(Rhinoclemmys nasuta, Testudines: Geoemydidae)

en un ambiente insular del Pacifico colombiano.

Caldasia 34(1): 109-125.

Giraldo A, Garcés-Restrepo MF, Carr JL. 2012c.

Kinosternon leucostomum (Duméril y_ Bibron).

Pp. 332-339 In: V. Biologia y Conservacion de las

Tortugas Continentales de Colombia. Serie Editorial

Recursos Hidrobiologicos y Pesqueros Continentales

de Colombia. Editors, Paez VP, Morales-Betancourt

MA, Lasso CA, Castafio-Mora OV, Bock BC. Instituto

de Investigacién de Recursos Biologicos Alexander

von Humboldt, Bogota, Colombia. 523 p.

Giraldo A, Garcés-Restrepo MF, Carr JL. 2013a.

Rhinoclemmys annulata (Gray, 1860). Catalogo de

Anfibios y Reptiles de Colombia 1(1): 31-34.

Giraldo A, Garcés-Restrepo MF, Carr JL. 2013b.

Kinosternon leucostomum (Duméril y Bibron en

Dumeril y Duméril, 1851). Catalogo de Anfibios y

Reptiles de Colombia 1(1): 45-49.

Gonzalez-Avendafio LP. 2019. Correlacién entre grupos

genéticos y variaciones bioclimaticas en la especie

Amphib. Reptile Conserv.

de tortuga continental, Rhinoclemmys melanosterna

(Testudines: Geoemydidae). Undergraduate Thesis,

Departamento de Biologia, Universidad Militar

Nueva Granada, Bogota, Colombia. 32 p.

Henao LD, Ruiz-Palma J. 1996. Estado actual de la

tortuga bache (Chelydra serpentina acutirostris) en el

Valle de Cauca: caceria y cautiverio en la comunidad

de San Cipriano, Buenaventura. Pp. 157—160 In:

Investigaciény Manejo de Fauna parala Construccion

de Sistemas Sostenibles. Editor, Osorio de la Cruz H.

Centro para la Investigacion en Sistemas Sostenibles

de Produccion Agropecuaria, Cali, Colombia. 239 p.

Heppell SS. 1998. Application of life-history theory

and population model analysis to turtle conservation.

Copeia 1998(2): 367-375.

Hernandez-Obando LS. 2020. Caracterizacion de la playa

Los Cocos sector Mendihuaca-Magdalena durante el

seguimiento de la temporada de anidacion de tortugas

marinas 2019. Undergraduate Thesis, Universidad

Jorge Tadeo Lozano, Programa de Biologia Marina,

Santa Marta, Magdalena, Colombia. 102 p.

HerpNET. 2021. HerpNET Data Portal. Available: https//

ecologicaldata.org/wiki/herpnet [Accessed: 3 January

2021].

Hurtado-Sepulveda SN. 1973. Algunos Aspectos

Bioecolégicos de Podocnemis lewyana (Dumeril,

1984) (Testudinata: Pleurodira: Pelomedusidae

1830). Biologia 1. Centro de _ Investigaciones

Biologicas Pesqueras del Rio Magdalena, Caldas,

Colombia. 34 p.

ICMBio (Instituto Chico Mendes de Conservacao da

Biodiversidade). 2018. Livro Vermelho da Fauna

Brasileira Ameacgada de Extingdo. Volume IV.

Répteis. Instituto Chico Mendes de Conservacgao

da Biodiversidade/Ministério do Meio Ambiente,

Brasilia, DF, Brazil. 253 p.

IDEAM. 2013. Zonificacion y Codificacién de Unidades

Hidrograficas e Hidrogeoldgicas de Colombia.

Instituto de Hidrologia, Meteorologia y Estudios

Ambientales, Bogota, Colombia. 46 p.

IGAC. 2021. Instituto de Geografia Agustin Codazzi

website. Available: http://igac.gov.co [Accessed: 3

January 2021].

Thlow F, Dambach J, Engler JO, Flecks M, Hartmann T,

Nekum S, Rajaei H, R6dder D. 2012. On the brink

of extinction? How climate change may affect global

chelonian species richness and distribution. Global

Change Biology 18(5): 1,520—1,530.

iNaturalist. 2021. iNaturalist website. Available: https://

www. inaturalist.org [Accessed: 3 January 2021].

Insacco G, Spadola F. 2010. First record of Kemp’s Ridley

Sea Turtle, Lepidochelys kempii (Garman, 1880)

(Chelontidae), from the Italian waters (Mediterranean

Sea). Acta Herpetologica 5(1): 113-117.

IUCN (international Union for the Conservation of

Nature). 2012. Guidelines for Application of the IUCN

Red List Criteria at Regional and National Levels:

April 2022 | Volume 16 | Number 1 | e306

Paez et al.

Version 4.0. International Union for the Conservation

of Nature, Gland, Switzerland. 41 p.

IUCN (international Union for the Conservation of

Nature). 2013. Guidelines for Reintroductions and

Other Conservation Translocations. Version 1.0.

International Union for the Conservation of Nature,

Gland, Switzerland. 57 p.

IUCN (international Union for the Conservation of

Nature). 2018. The IUCN Red List of Threatened

Species. Version 2018-2. Available at: http://www.

iucnredlist.org [Accessed: 3 January 2021].

Iverson JB. 1992. A Revised Checklist with Distribution

Maps of the Turtles of the World. Privately printed,

Richmond, Indiana, USA. 363 p.

Jackson JT, Starkey DE, Guthrie RW, Forstner MRJ.

2008. A mitochondrial DNA phylogeny of extant

species of the genus TJrachemys with resulting

taxonomic implications. Chelonian Conservation and

Biology 7(1): 131-135.

Jensen EL, Govindarajulu P, Russello MA. 2014. When

the shoe doesn’t fit: applying conservation unit

concepts to Western Painted Turtles at their northern

periphery. Conservation Genetics 15(2): 261-274.

Karl SS, Bowen BW. 1999. Evolutionary significant units

versus geopolitical taxonomy: molecular systematics

of an endangered sea turtle (genus Chelonia).

Conservation Biology 13(5): 990-999.

Lasso CA, Trujillo F, Morales-Betancourt MA, Amaya

L, Caballero S, Castafieda B. 2018. Conservacion

y trafico de la tortuga matamata, Chelus fimbriata

(Schneider, 1783) en Colombia: un ejemplo del

trabajo conjunto entre el Sistema Nacional Ambiental,

ONG y academia. Biota Colombiana 19(1): 147-159.

Lasso CA, Trujillo F, Morales-Betancourt MA. (Editors).

2020. VIII. Biodiversidad de la Reserva Natural

Bojonawi, Vichada, Colombia: rio Orinoco y Planicie

de Inundacion. Serie Editorial Fauna_ Silvestre

Neotropical. Instituto de Investigacion de Recursos

Biologicos Alexander von Humboldt. Bogota,

Colombia. 556 p.

Laurance W, Useche DC, Rendeiro J, Kalka M, Bradshaw

CJ, Sloan SP, Laurance SG, Campbell M, Abernethy

K, Zamzani F. 2012. Averting biodiversity collapse

in tropical forest protected areas. Nature 489(7415).:

290-294.

Lehn C, Das I, Forstner MRJ, Brown RM. 2007.

Responsible vouchering in turtle research: an

introduction and recommendations. Chelonian

Research Monographs 4: 147-156.

Leviton AE, Gibbs RH, Dawson CE. 1985. Standards

in herpetology and ichthyology. Part I. Standard

symbolic codes for institution resource collections in

herpetology and ichthyology. Copeia 1985(3): 802—

832.

Liévano-Latorre LF, Brum FT, Loyola R. 2021. How

effective have been guerrilla occupation and protected

areas in avoiding deforestation in Colombia?

Amphib. Reptile Conserv.

Biological Conservation 253: 108916.

Lovich JE, Ennen JR, Agha M, Gibbons JW.

2018. Where have all the turtles gone, and why does it

matter? BioScience 68(10): 771-781.

MADS (Ministerio de Ambiente y Desarrollo Sostenible).

2015. Decreto 1076 del 26 de mayo de 2015 [con

fuerza de ley]. Por medio del cual se expide el

Decreto Unico Reglamentario del Sector Ambiente y

Desarrollo Sostenible. 26 de mayo de 2015. D.O. No.

49523. Available: https:/(www.minambiente.gov.co/

images/normativa/decretos/2015/DECRETO_1076__

DEL_26 DE MAYO DE 2015.pdf [Accessed: 3

January 2021].

MADS (Ministerio de Ambiente y Desarrollo Sostenible).

2016. Decreto 1272 del 03 de agosto de 2016 [con

fuerza de ley]. Por el cual se adiciona un capitulo al

Titulo 9 de la Parte 2 del Libro 2 del Decreto 1076

de 2015, Decreto Unico Reglamentario del Sector

Ambiente y Desarrollo Sostenible, en lo relacionado

con la tasa compensatoria por caza de fauna

silvestre y se dictan otras disposiciones. Available:

http://es.presidencia. gov.co/normativa/normativa/

DECRETO0%201272%20DEL%2003%20DE%20

AGOSTO%20DE%202016.pdf [Accessed: 3 January

2021].

Marquez R. 1990. FAO species catalogue, Volume 11.

Sea turtles of the world: an annotated and illustrated

catalogue of sea turtle species known to date. FAO

Fisheries Synopsis 125(11): 1-81.

Martinez-Callejas SJ, Duran-Prieto C, Paez VP, Trujillo

F, Trujillo-Pérez A. 2015. Charapa. Podocnemis

expansa (Miller, 1935). Pp. 132-136 In: Libro Rojo

de Reptiles de Colombia (2015). Editors, Morales-

Betancourt MA, Lasso CA, Paez VP, Bock BC.

Instituto de Investigacién de Recursos Biologicos

Alexander von Humboldt, Bogota, Colombia. 258 p.

Martinez-Ortega J, MHernandez-Fernandez J. 2013.

Método Taguchi para optimizar marcadores RAPD-

PCR y determinar diversidad genética: un modelo,

la tortuga cabezona Caretta caretta (Testudines:

Cheloniidae). Mutis 3(1): 49-68.

McCord WP, Joseph-Ouni M, Lamar WW. 2001. A

taxonomic reevaluation of Phyrnops (Testudines:

Chelidae) with the description of two new genera and

a new species of Batrachemys. Revista de Biologia

Tropical 49(2): 715-764.

McCord WP, Joseph-Ouni M, Hagen C, Blank T.

2010. Three new subspecies of 7rachemys venusta

(Testudines: Emydidae) from Honduras, northern

Yucatan (Mexico), and Pacific coastal Panama.

Reptilia (GB) (Barcelona) 71: 39-49.

McCranie JR. 2018. The lizards, crocodiles, and

turtles of Honduras. Systematics, distribution, and

conservation. Bulletin of the Museum of Comparative

Zoology, Special Publications Series 2: 1-666.

Medem F. 1958. Informe sobre reptiles colombianos

(II). El conocimiento actual sobre la distribucién

April 2022 | Volume 16 | Number 1 | e306

Turtles of Colombia: diversity, distribution, and conservation

geografica de las Testudinata en Colombia. Boletin del

Museo de Ciencias Naturales, Caracas 2-3: 13-45.

Medem F. 1960. Datos zoogeograficos y ecolégicos sobre

los Crocodylia y Testudinata de los rios Amazonas,

Putumayo y Caqueta. Caldasia 8(38): 341-351.

Medem F. 1962. La distribucién geografica de los

Crocodylia y Testudinata en el departamento del

Choco. Revista de la Academia Colombiana de

Ciencias Exactas, Fisicas, y Naturales 11(44): 279-

303.

Medem F. 1968. El desarrollo de la herpetologia en

Colombia. Revista de la Academia Colombiana de

Ciencias Exactas, Fisicas y Naturales 13(50): 160—

163.

Medem F. 1969. Estudios adicionales sobre los Crocodylia

y Testudinata del Alto Caqueta y rio Caguan. Caldasia

10(48): 329-353.

Medem F. 1977. Contribucion al conocimiento sobre la

taxonomia, distribucion geografica y ecologia de la

tortuga “Bache” (Chelydra serpentina acutirostris).

Caldasia 12(56): 41-101.

Medem F. 1983. Reproductive data on Platemys

platycephala (Testudines: Chelidae) in Colombia.

Pp. 249-234 In: Advances in Herpetology and

Evolutionary Biology. Editors, Rhodin AGJ, Miyata

K. Museum of Comparative Zoology, Harvard

University, Cambridge, Massachusetts, USA. 725 p.

Medina-Rangel GF. 2011. Diversidad alfa y beta de la

comunidad de reptiles en el complejo cenagoso de

Zapatosa, Colombia. Revista de Biologia Tropical

59(2): 935-968.

Medina-Rangel GF, Cardenas-Arevalo G. 2015.

Relaciones espaciales y alimenticias del ensamblaje

de reptiles del complejo cenagoso de Zapatosa,

Departamento del Cesar (Colombia). Papéis Avulsos

de Zoologia 55(10): 143-165.

Molina FB, Machado FA, Zaher H. 2012. Taxonomic

validity of Mesoclemmys heliostemma (McCord,

Joseph-Ouni, and Lamar, 2001) (Testudines: Chelidae)

inferred from morphological analysis. Zootaxa 3575:

63-77.

Moll EO, Legler JM. 1971. The life history ofa neotropical

slider turtle, Pseudemys scripta (Schoepff), in

Panama. Bulletin of the Los Angeles County Museum

of Natural History 11: 1-102.

Moncada F, Romero S. 2015. Nota sobre los registros

del género Lepidochelys (Cryptodira: Cheloniidae) en

Cuba. Solenodon 12: 161-163.

Moncada F, Ramirez-Gallego C, Camero J, Gonzalez M,

Nodarse G, Barrientos-Mufioz KG. 2019. A juvenile

Loggerhead Turtle (Caretta caretta) tagged in Cuba

is recaptured in Colombian waters. Marine Turtle

Newsletter 156: 30-32.

Montes-Correa AC, Saboya-Acosta LP, Paez VP, Vega

K, Renjifo JM. 2014. Distribucioén de tortugas

continentales del Caribe colombiano. Acta Bioldgica

Colombiana 19(3): 341-350.

Amphib. Reptile Conserv.

Montes-Correa AC, Saboya-Acosta LP, Paez VP, Renyjifo

JM. 2017. Kinosternon scorpioides (Linnaeus, 1766).

Catalogo de Anfibios y Reptiles de Colombia 3(1).:

17-24.

Morales-Betancourt MA, Lasso CA. 2012a.

Rhinoclemmys diademata (Mertens, 1954). Pp. 305-

307 In: V. Biologia y Conservacion de las Tortugas

Continentales de Colombia. Serie Editorial Recursos

Hidrobiologicos y Pesqueros Continentales de

Colombia. Editors, Paez VP, Morales-Betancourt MA,

Lasso CA, Castafio-Mora OV, Bock BC. Instituto de

Investigacion de Recursos Biologicos Alexander von

Humboldt, Bogota, Colombia. 523 p.

Morales-Betancourt MA, Lasso CA. 2012b. Chelus

fimbriatus (Schneider, 1783). Pp. 243-246 In:

V. Biologia y Conservacién de las Tortugas

Continentales de Colombia. Serie Editorial Recursos

Hidrobiolégicos y Pesqueros Continentales de

Colombia. Editors, Paez VP, Morales-Betancourt MA,

Lasso CA, Castafio-Mora OV, Bock BC. Instituto de

Investigacion de Recursos Biologicos Alexander von

Humboldt, Bogota, Colombia. 523 p.

Morales-Betancourt MA, Lasso CA. 2012c.

Mesoclemmys gibba (Schweigger, 1812). Pp. 254—

256 In: V. Biologia y Conservacion de las Tortugas

Continentales de Colombia. Serie Editorial Recursos

Hidrobiolégicos y Pesqueros Continentales de

Colombia. Editors, Paez VP, Morales-Betancourt MA,

Lasso CA, Castafio-Mora OV, Bock BC. Instituto de

Investigacion de Recursos Biologicos Alexander von

Humboldt, Bogota, Colombia. 523 p.

Morales-Betancourt MA, Lasso CA. 2012d.

Mesoclemmys raniceps (Gray, 1856). Pp. 259-260

In: V. Biologia y Conservacion de las Tortugas

Continentales de Colombia. Serie Editorial Recursos

Hidrobiolégicos y Pesqueros Continentales de

Colombia. Editors, Paez VP, Morales-Betancourt MA,

Lasso CA, Castafio-Mora OV, Bock BC. Instituto de

Investigacion de Recursos Biologicos Alexander von

Humboldt, Bogota, Colombia. 523 p.

Morales-Betancourt MA, Lasso CA. 2015a. Conservacion

de grandes reptiles acuaticos continentales (tortugas

y crocodilidos) en Colombia. Pp. 53-70 In:

Conservacion de Grandes Vertebrados en Areas no

Protegidas de Colombia, Venezuela y Brasil. Serie

Editorial Fauna_ Silvestre Neotropical. Editors,

Payan E, Lasso CA, Castafio-Uribe E. Instituto de

Investigacion de Recursos Biologicos Alexander von

Humboldt, Bogota, Colombia. 294 p.

Morales-Betancourt MA, Lasso CA. 2015b. Inguensa.

Rhinoclemmys diademata (Mertens, 1954). Pp. 146-

148 In: Libro Rojo de Reptiles de Colombia (2015).

Editors, Morales-Betancourt MA, Lasso CA, Paez

VP, Bock BC. Instituto de Investigacion de Recursos

Biologicos Alexander von Humboldt, Bogota,

Colombia. 258 p.

Morales-Betancourt MA, Lasso CA. 2015c. Chipiro.

April 2022 | Volume 16 | Number 1 | e306

Paez et al.

Podocnemis erythrocephala (Spix, 1824). Pp. 158-

160 In: Libro Rojo de Reptiles de Colombia (2015).

Editors, Morales-Betancourt MA, Lasso CA, Paez

VP, Bock BC. Instituto de Investigacion de Recursos

Biologicos Alexander von Humboldt, Bogota,

Colombia. 258 p.

Morales-Betancourt MA, Lasso CA, Trujillo F, de la

Ossa J, Forero G, Paez VP. 2012a. Amenazas a las

poblaciones de tortugas continentales de Colombia.

Pp. 453-492 In: V. Biologia y Conservacion de las

Tortugas Continentales de Colombia. Serie Editorial

Recursos Hidrobiologicos y Pesqueros Continentales

de Colombia. Editors, Paez VP, Morales-Betancourt

MA, Lasso CA, Castafio-Mora OV, Bock BC. Instituto

de Investigacién de Recursos Biologicos Alexander

von Humboldt, Bogota, Colombia. 523 p.

Morales-Betancourt MA, Lasso CA, Paez VP. 2012b.

Rhinemys rufipes (Spix, 1824). Pp. 269-271

In: V. Biologia y Conservacion de las Tortugas

Continentales de Colombia. Serie Editorial Recursos

Hidrobiologicos y Pesqueros Continentales de

Colombia. Editors, Paez VP, Morales-Betancourt MA,

Lasso CA, Castafio-Mora OV, Bock BC. Instituto de

Investigacion de Recursos Biologicos Alexander von

Humboldt, Bogota, Colombia. 523 p.

Morales-Betancourt MA, Lasso CA, Paez VP. 2012c.

Phrynops geofjroanus (Schweigeger, 1812). Pp. 266—

268 In: V. Biologia y Conservacion de las Tortugas

Continentales de Colombia. Serie Editorial Recursos

Hidrobiologicos y Pesqueros Continentales de

Colombia. Editors, Paez VP, Morales-Betancourt MA,

Lasso CA, Castafio-Mora OV, Bock BC. Instituto de

Investigacion de Recursos Biologicos Alexander von

Humboldt, Bogota, Colombia. 523 p.

Morales-Betancourt MA, Lasso CA, Trujillo F.

2012d. Podocnemis vogli (Miller). Pp. 399-403

In: V. Biologia y Conservacioén de las Tortugas

Continentales de Colombia. Serie Editorial Recursos

Hidrobiol6gicos y Pesqueros Continentales de

Colombia. Editors, Paez VP, Morales-Betancourt MA,

Lasso CA, Castafio-Mora OV, Bock BC. Instituto de

Investigacion de Recursos Biologicos Alexander von

Humboldt, Bogota, Colombia. 523 p.

Morales-Betancourt MA, Munera C, Lasso CA. 2012e.

Trachemys scripta elegans (Wied, 1838). Pp. 252—255

In: VI. Catalogo de la Biodiversidad Acudtica Exotica

y Transplantada en Colombia: Moluscos, Crustdceos,

Peces, Anfibios, Reptiles y Aves. Serie Editorial

Recursos Hidrobiologicos y Pesqueros Continentales

de Colombia. Editors, Gutiérrez F de P, Lasso CA,

Baptiste MP, Sanchez-Duarte P, Diaz AM. Instituto de

Investigacion de los Recursos Biolégicos Alexander

von Humboldt, Bogota, Colombia. 170 p.

Morales-Betancourt MA, Lasso CA, Paez VP, Bock BC.

2015a. Libro Rojo de Reptiles de Colombia (2015).

Instituto de Investigacion de Recursos Bioldgicos

Alexander von Humboldt (I[AvH), Bogota, Colombia.

Amphib. Reptile Conserv.

131

258 p.

Morales-Betancourt MA, Lasso CA, Paez VP. 2015b.

Terecay. Podocnemis unifilis (Troschel, 1848). Pp.

149-152 In: Libro Rojo de Reptiles de Colombia

(2015). Editors, Morales-Betancourt MA, Lasso

CA, Paez VP, Bock BC. Instituto de Investigacion

de Recursos Biologicos Alexander von Humboldt,

Bogota, Colombia. 258 p.

Moreno R, Negrete E. 2012. Marco normativo de la

fauna silvestre en Colombia con fines de seguridad

alimentaria y consumo de subsistencia. Pp. 32-63

In: Carne de Monte y Seguridad Alimentaria: Bases

Técnicas para una Gestion Integral en Colombia.

Editor, Restrepo S. Instituto de Investigacion de

Recursos Bioldgicos Alexander von Humboldt,

Bogota, Colombia. 94 p.

Moreno-Munar A, Ospina-Sanchez SC, Jauregui-

Romero GA, Alvarez-Leon R. 2014. Monitoreamiento

de poblaciones de tortugas marinas en los sectores

de Arrecifes y Cafiaveral, Parque Nacional Natural

Tayrona, Colombia. Arquivos de Ciéncias do Mar

47(2): 19-30.

Muller L. 1939. Uber die Verbreitungdel Chelonier auf

dem Sudamerikanischen Kontinent. Physis 16: 89-

102.

Negret PJ, Allan J, Braczkowski A, Maron M, Watson

JEM. 2017. Need for conservation planning in

postconflict Colombia. Conservation Biology 31(3):

499-500.

Oksanen J, Blanchet FG, Friendly M, Kindt R, Legendre

P, McGlinn D, Minchin PR, O’ Hara RB, Wagner H,

et al. 2007. Vegan: Community Ecology Package. R

package version 24-0. Available: https://cran.r-project.

org/web/packages/vegan/index.html [Accessed: 3

January 2021].

Ortiz-Yusty C, Restrepo A, Paez VP. 2014. Distribucion

potencial de Podocnemis lewyana_ (Reptilia:

Podocnemididae) y su posible fluctuacién bajo

escenarios de cambio climatico global. Acta Bioldgica

Colombiana 19(3): 471481.

Paez VP. 2015. Tortuga del rio Magdalena. Podocnemis

lewyana (Dumeéril, 1852). Pp. 137—140 In: Libro Rojo

de Reptiles de Colombia (2015). Editors, Morales-

Betancourt MA, Lasso CA, Paez VP, Bock BC.

Instituto de Investigaci6n de Recursos Biologicos

Alexander von Humboldt, Bogota, Colombia. 258 p.

Paez VP, Bock BC, Estrada JJ, Ortega AM, Daza JM,

Gutiérrez-Cardenas PD. 2002. Guia de Campo de

algunas Especies de Anfibios y Reptiles de Antioquia.

Multilmpresos Ltda., Medellin, Colombia. 136 p.

Paez VP, Morales-Betancourt MA, Lasso CA, Castafio-

Mora OV, Bock BC. (Editors). 2012a. V. Biologia

y Conservacion de las Tortugas Continentales de

Colombia. Serie Editorial Recursos Hidrobiologicos

y Pesqueros Continentales de Colombia. Instituto de

Investigacion de los Recursos Biolégicos Alexander

von Humboldt, Bogota, Colombia. 523 p.

April 2022 | Volume 16 | Number 1 | e306

Turtles of Colombia: diversity, distribution, and conservation

Paez VP, Restrepo-Isaza A, Vargas-Ramirez M, Bock

BC, Gallego-Garcia N. 2012b. Podocnemis lewyana

(Dumeril, 1852). Pp. 375-381 In: VK Biologia y

Conservacion de las Tortugas Continentales de

Colombia. Serie Editorial Recursos Hidrobiologicos

y Pesqueros Continentales de Colombia. Editors, Paez

VP, Morales-Betancourt MA, Lasso CA, Castafio-

Mora OV, Bock BC. Instituto de Investigacion de

Recursos Bioldgicos Alexander von Humboldt,

Bogota, Colombia. 523 p.

Paez VP, Restrepo A, Vargas-Ramirez M, Bock BC,

Gallego-Garcia N. 2013. Podocnemis lewyana

(Dumeril, 1852). Catdlogo de Anfibios y Reptiles de

Colombia 1(1): 1-5.

Paez VP, Ramirez-Gallego C, Barrientos-Mufioz KG.

2015a. Caguama. Caretta caretta (Linnaeus, 1758).

Pp. 118-121 In: Libro Rojo de Reptiles de Colombia

(2015). Editors, Morales-Betancourt MA, Lasso

CA, Paez VP, Bock BC. Instituto de Investigacion

de Recursos Biolégicos Alexander von Humboldt,

Bogota, Colombia. 258 p.

Paez VP, Ramirez-Gallego C, Barrientos-Mufioz KG.

2015b. Tortuga verde. Chelonia mydas (Linnaeus,

1758). Pp. 153-156 In: Libro Rojo de Reptiles de

Colombia (2015). Editors, Morales-Betancourt

MA, Lasso CA, Paez VP, Bock BC. Instituto de

Investigacion de Recursos Biologicos Alexander von

Humboldt, Bogota, Colombia. 258 p.

Parham JF, Papenfuss TJ, Buskirk JR, Parra-Olea G,

Chen J, Simison WB. 2015. Trachemys ornata or

not ornata: reassessment of a taxonomic revision for

Mexican Trachemys. Proceedings of the California

Academy of Science 62: 359-367.

Patifio-Martinez J, Marco A, Quifiones L, Godley BJ.

2008. Globally significant nesting of the Leatherback

Turtle (Dermochelys coriacea) on the Caribbean coast

of Colombia and Panama. Biological Conservation

141(8): 1,982-1,988.

Pérez J, Alegria J. 2009. Evaluacion morfométrica

y dimorfismo sexual _intra-poblacional de

Rhinoclemmys nasuta (Boulenger, 1902) en una zona

insular continental del Pacifico colombiano. Revista

Colombiana de Ciencia Animal 1(2): 143-156.

Pinto-Erazo MA, Calderén-Espinosa ML, Medina-

Rangel GF, Méndez-Galeano MA. 2020. Herpetofauna

from two municipalities of southwestern Colombia.

Biota Colombiana 21(1): 41-57.

Pritchard PCH. 2008. Chelus fimbriata (Schneider 1783),

Matamata Turtle. Chelonian Research Monographs 5:

020.1-020.10.

Pritchard PCH, Trebbau P. 1984. The Turtles of Venezuela.

Society for the Study of Amphibians and Reptiles,

Oxford, Ohio, USA. 352 p.

R Core Team. 2016. R: a language and environment for

statistical computing. R Foundation for Statistical

Computing, Vienna, Austria. Available: https://

www.R-project.org [Accessed: 3 January 2021].

Amphib. Reptile Conserv.

Ramirez-Gallego C, Barrientos-Mufioz KG, Paez VP.

2015. Dermochelys coriacea (Vandelli, 1761). Pp.

122-126 In: Libro Rojo de Reptiles de Colombia

(2015). Editors, Morales-Betancourt MA, Lasso

CA, Paez VP, Bock BC. Instituto de Investigacion

de Recursos Biologicos Alexander von Humboldt,

Bogota, Colombia. 258 p.

Ramirez-Gallego C, Barrientos-Mufioz KG. 2020a.

Sea turtles at Serrana Island and Serranilla Island,

Seaflower Biosphere Reserve, Colombian Caribbean.

Frontiers in Marine Science 6: \—S.

Ramirez-Gallego C, Barrientos-Mufioz KG. 2020b.

Illegal Hawksbill trafficking: five years of records

of the handicrafts and meat trades of the Hawksbill

Turtle (Eretmochelys imbricata) in Cartagena de

Indias, Colombia. Journal of Aquaculture and Marine

Biology 9(3): 101-105.

Ramirez-Gallego C, Barrientos-Mufioz KG. 2021.

Stopping illegal Hawksbill trafficking in Cartagena

de Indias, Colombia. Pp. 141-148 In: Sea Turtle

Research and Conservation: Lessons from Working in

the Field. Editor, Nahill B. Elsevier Academic Press,

Cambridge, Massachusetts, USA. 224 p.

Regalado-Tabares AC, Botero-Botero A, Mufiera-

Isaza C, Ortega-Guio AF, Restrepo-Isaza A. 2012.

Chelydra acutirostris. Pp. 275-278 In: V. Biologia

y Conservacion de las Tortugas Continentales de

Colombia. Serie Editorial Recursos Hidrobiologicos

y Pesqueros Continentales de Colombia. Editors, Paez

VP, Morales-Betancourt MA, Lasso CA, Castafio-

Mora OV, Bock BC. Instituto de Investigacion de

Recursos Bioldgicos Alexander von Humboldt,

Bogota, Colombia. 523 p.

Renjifo JM, Acosta-Galvis R. 2014. Anfibios y

reptiles de la Estrella Fluvial Inirida. Pp. 131-153

In: Biodiversidad de la Estrella Fluvial Inirida.

Editors, Trujillo F Usma JS, Lasso CA. Instituto de

Investigacion de Recursos Biologicos Alexander von

Humboldt, Bogota, Colombia. 328 p.

Renjifo JM, Lasso CA, Morales-Betancourt MA. 2009.

Herpetofauna de la estrella fluvial de Inirida (rios

Inirida, Guaviare, Atabapo, y Orinoco), Orinoquia

colombiana: lista preliminar de especies. Biota

Colombiana 10(1—2): 171-178.

Renteria-Moreno LE, Forero-Medina G, Garcés-

Restrepo MF, Carr JL, Rueda Almonacid JV. 2012.

Range extension of Kinosternon dunni Schmidt,

1947 (Reptilia, Testudines, Kinosternidae) in Choco,

Colombia. Check List 8(6): 1,310—1,312.

Restrepo-Bastidas E, Menoza-Mora A. 2018. Guia

Ilustrada de Anfibios y Reptiles de la Ciénaga

de Palagua, Boyacad, Colombia. Centro Pecuario

y Agroempresarial SENA, La Dorada, Caldas,

Colombia. 59 p.

Restrepo A, Paez VP, Bock BC, Daza JM, Diazgranados

I. 2014. Trachemys callirostris (Gray, 1856). Catalogo

de Anfibios y Reptiles de Colombia 2(2). 7-12.

April 2022 | Volume 16 | Number 1 | e306

Paez et al.

Ripple WJ, Wolf C, Newsome TM, Hoffman M, Wirsing

AJ, McCauley DJ. 2017. Extinction risk is most

acute for the world’s largest and smallest vertebrates.

Proceedings of the National Academy of Sciences of

the United States of America 114(40): 10,678—10,683.

Rivera-Gomez M, Calle-Bonilla IC, Cuéllar-Chacon A,

Paredes-Mina F, Giraldo-Lopez A. 2016. Registro de

la tortuga cana (Dermochelys coriacea) en el Parque

Nacional Natural Gorgona, Pacifico colombiano.

Biota Colombiana 17(2): 163-166.

Rivera-Robles DL, Adames-Jiménez TA. 2021.

Lepidochelys olivacea (Eschsholtz, 1829). Catalogo

de Anfibios y Reptiles de Colombia 7(1): 30-38.

Rodriguez JP, Garcia-Rawlins A, Rojas-Suarez F.

(Editors). 2015. Libro Rojo de la Fauna Venezolana.

Provita y Fundacion Empresas Polar, Caracas,

Venezuela. 287 p.

Rodriguez-Murcia JD, Giraldo A, Garcés-Restrepo MF,

Sanchez F. 2014. Estructura poblacional y dimorfismo

sexual de Kinosternon leucostomum (Testudines:

Kinosternidae) en un sistema de charcas asociadas

al rio Purnid, Caldas, Colombia. Jnvestigacion,

Biodiversidad y Desarrollo 33(2): 86-95.

RUNAP. 2018. Registro Unico de Areas Protegidas.

Available: http://runap.parquesnacionales.gov.co/

cifras [Accessed: 3 January 2021].

Sanchez-Villagra MR, Pritchard PCH, Paolillo A,

Linares OJ. 1995. Geographic variation in the

Matamata turtle, Chelus fimbriatus, with observations

on its shell morphology and morphometry. Chelonian

Conservation and Biology 1(4): 293-300.

Seidel ME. 2002. Taxonomic observations on extant

species and subspecies of slider turtles, genus

Trachemys. Journal of Herpetology 36(2): 285-292.

Sepulveda-Seguro AM, Paez VP, Gomez-Rincon

MA, Morales-Betancourt MA, Lasso CA. 2020.

Assessment of population characteristics and habitat

use of Podocnemis vogli (Muller, 1935) during a

reproductive season at the Bojonawi Private Nature

Reserve (Vichada Department, Colombia). Revista

Latinoamericana de Herpetologia 3(2): 49-60.

SERFOR. 2018. Libro Rojo de la Fauna Silvestre

Amenazada del Peru. I Edicion. Servicio Nacional

Forestal y de Fauna Silvestre, Lima, Peru. 548 p.

SIAC. 2021. Sistema de Informacioén Ambiental de

Colombia. Available: http//siac.gov.co [Accessed: 3

January 2021].

SiB Colombia. 2021. Sistema de Informacién sobre

Biodiversidad de Colombia. Available: — http://

sibcolombia.net [Accessed: 3 January 2021].

Sinovas P, Price B, King E, Hinsley A, Pavitt A.

2017. Comercio de Vida Silvestre en los Paises

Amazonicos: Andlisis del Comercio de _ Especies

Listadas en CITES. Informe Técnico Preparado para

el Programa Regional Amazonia (BMZ/DGIS/GIZ).

UN Environment, World Conservation Monitoring

Centre, Cambridge, United Kingdom. 116 p.

Amphib. Reptile Conserv.

Stephens PR, Weins JJ. 2003. Ecological diversification

and phylogeny of emydid turtles. Biological Journal

of the Linnean Society 79(4): 577-610.

Tiago P, Ceia-Hasse A, Marques TA, Capinha C, Pereira

HM. 2017. Spatial distribution of citizen science

casuistic observations for different taxonomic groups.

Scientific Reports 7: 12832.

Tobon-Lopez A, Amorocho DF. 2014. Estudio

poblacional de la tortuga carey Eretmochelys imbricata

(Chelontidae) en el Pacifico sur de Colombia. Acta

Biologica Colombiana 19(3): 447-459.

Torres-Carvajal O, Pazmifio-Otamendi G, Salazar-

Valenzuela D. 2019. Reptiles of Ecuador: a

resource-rich online portal, with dynamic checklists

and photographic guides. Amphibian & Reptile

Conservation 13(1) [General Section]: 209-229

(e178).

Trujillo-Arias N, Amorocho DF, Lopez-Alvarez D,

Mejia-Ladino LM. 2014. Phylogeographic relations

of some feeding and nesting of Hawksbill Turtle

rookeries (Eretmochelys imbricata) in the Caribbean

and Pacific of Colombia. Boletin de Investigaciones

Marinas y Costeras 43(1): 159-182.

TTWG — Turtle Taxonomy Working Group; Rhodin

AGJ, Iverson JB, Bour R, Fritz U, Georges A,

Shaffer HB, van Dijk PP. 2017. Turtles of the world:

annotated checklist and atlas of taxonomy, synonymy,

distribution, and conservation status (8" Edition).

Chelonian Research Monographs 7: \—292.

TTWG — Turtle Taxonomy Working Group; Rhodin

AGJ, Iverson JB, Bour R, Fritz U, Georges A,

Shaffer HB, van Dijk PP. 2021. Turtles of the world:

Annotated checklist and atlas of taxonomy, synonymy,

distribution, and conservation status (9" Edition).

Chelonian Research Monographs 8: \—472.

Uetz P, Freed P, Aguilar R, HoSek J. (Editors). 2021.

The Reptile Database. Available: http://www.reptile-

database.org [Accessed: 3 January 2021].

Vargas-Ramirez M, Maran J, Fritz U. 2010. Red- and

Yellow-footed Tortoises, Chelonoidis carbonaria and

C. denticulata (Reptilia: Testudines: Testudinidae),

in South America’s savannahs and forests: Do their

phylogeographies reflect distinct habitats? Organisms

Diversity and Evolution 10(2): 161-172.

Vargas-Ramirez M, Carr JL, Fritz U. 2013. Complex

phylogeography in Rhinoclemmys melanosterna:

conflicting mitochondrial and nuclear evidence

suggests past hybridization (Testudines:

Geoemydidae). Zootaxa 3670(2): 238-254.

Vargas-Ramirez M, del Valle C, Ceballos-Fonseca

CP, Fritz U. 2017. Trachemys medemi n. sp. from

northwestern Colombia turns the biogeography of

South American slider turtles upside down. Journal

of Zoological Systematics and Evolutionary Research

55: 326-339.

Vargas-Ramirez M, Caballero S, Morales-Betancourt

MA, Lasso CA, Amaya L, Martinez JG, Silva-Viana

April 2022 | Volume 16 | Number 1 | e306

Turtles of Colombia: diversity, distribution, and conservation

M das N, Vogt RC, Pires-Farias I, Hrbek T, Fritz U,

et al. 2020. Genomic analyses reveal two species of

the Matamata (Testudines: Chelidae: Chelus spp.) and

clarify their phylogeny. Molecular Phylogenetics and

Evolution 148: 106823.

Vasquez-Carillo C, Noriega-Hoyos CL, Hernandez-

Rivera L, Jauregui-Romero GA, Sullivan Sealey K.

2020. Genetic diversity and demographic connectivity

of Atlantic Green Sea Turtles at foraging grounds

in northeastern Colombia, Caribbean. Frontiers in

Marine Science 7(96): 1-12.

Wermuth H, Mertens R. 1961. Schildkréten, Krokodile,

Briickenechsen. Gustav Fisher Verlag, Jena, Germany.

A472 p.

Western D. 1989. Conservation without parks: wildlife in

the rural landscape. Pp. 158-165 In: Conservation for the

Twenty-first Century. Editors, Western D, Pearl M. Oxford

University Press, Oxford, United Kingdom. 365 p.

Williams EE. 1956. Pseudemys scripta callirostris from

Venezuela with a general survey of the scripta series.

Bulletin of the Museum of Comparative Zoology

115(5): 145-160.

Zapata-Ciro J, Guevara G, Castafio-Villa GJ. 2016.

Conocimiento popular y perspectivas de conservacion

sobre las tortugas continentales en la parte baja del

rio La Miel (Colombia). Revista Luna Azul 43: 15—28.

ZIMS. 2021. Zoological Information Management

System. Available: https://zims.species360.org/Login.

aspx [Accessed: 3 January 2021].

Vivian P. Paez obtained her Ph.D. degree in Ecology, Ethology, and Evolution from Ohio University

Photo by Jessica Bock Paez.

Alvaro Orozco.

Amphib. Reptile Conserv.

(Athens, Ohio, USA) in 1995, and is currently a Professor in the Instituto de Biologia of the

Universidad de Antioquia in Medellin, Colombia, where she teaches courses in Population Ecology

and Herpetology. She has edited two books and published over 70 scientific articles and book chapters.

Her research interests have focused on the influences of nest microclimatic conditions and paternal

effects on different fitness components of turtles with temperature-dependent sex determination. She

is also conducting demographic projects using population matrix models to permit the elaboration of

management plans for several species of freshwater turtles. Since arriving in Antioquia, Colombia,

she has been involved in several projects on the natural history and diversity of the herpetofauna in

this region, including the founding of the Museo de Herpetologia of the Universidad de Antioquia

(MHUA). Photo by Monica Nieto.

Brian C. Bock obtained his Ph.D. degree in Ethology from the University of Tennessee (Knoxville,

Tennessee, USA) in 1984. He held Smithsonian, Fulbright, and American Association for the

Advancement of Science (AAAS) fellowships before moving to Colombia, first as a Professor at the

Universidad Nacional de Colombia in Medellin, Colombia, and now as a Professor in the Instituto

de Biologia of the Universidad de Antioquia in Medellin, Colombia, where he teaches courses in

Conservation Biology and Behavioral Ecology. Brian has edited two books and published over 70

scientific articles and book chapters. His early research focused on how reptile movement patterns

influence population structure, but he has also conducted studies on reptile nesting ecology and

demography, as well as on the population genetics of other species of Colombian flora and fauna.

Diego A. Alzate-Estrada is a Biologist with a degree from the Universidad de Antioquia in Medellin,

Colombia, and a Master’s degree from the same university. He is interested in the population ecology

and conservation of freshwater turtles and crocodilians. Photo by Lucas Burgos Alvarez.

Karla Georgina Barrientos-Muifioz is a Biologist with a degree from the Universidad de

Antioquia in Medellin, Colombia, and a Master’s degree from the University of Puerto Rico,

Rio Piedras Campus in San Juan, Puerto Rico. She is a co-founder and scientific director of the

Fundacion Tortugas del Mar in Colombia and country coordinator in Colombia for the Wider

Caribbean Sea Turtle Conservation Network (WIDECAST). Karla has published over 13 scientific

articles and book chapters, and she was the winner of the Archie Carr Student Award - Runner-

Up: Biology, by the International Sea Turtle Society in 2014. Her research and interests are on

the nesting ecology and in-water assessments of sea turtles, and the trade of sea turtles, focusing

on the “tortoiseshell,” conservation biology, community outreach, and relationships between local

communities and stakeholders for applicable solutions to sea turtle conservation issues. Photo by

April 2022 | Volume 16 | Number 1 | e306

Amphib. Reptile Conserv.

Paez et al.

Viviana Cartagena graduated in Biology from the Universidad de Antioquia in Medellin,

Colombia, in 2013, and in 2020 received a Master’s degree in Biology from the same university. She

is interested in the population ecology and conservation of amphibians and reptiles. Photo by Laura

Cristina Osorno-Giraldo.

Andrea Echeverry-Alcendra graduated as a Biologist from the Universidad del Magdalena, Santa

Marta, Colombia, in 2009, and is completing her Master’s degree in Conservation and Biodiversity

at the Pontificia Universidad Javeriana, Bogota, Colombia. She works as Coordinator of Animal

Collections of the Barranquilla Zoo and is a member of the Conservation Planning Specialist Group

of the IUCN. Her research interests range across diverse topics in wildlife management, but with an

emphasis on the ecology and conservation of tortoises and freshwater turtles under the “One Plan

Approach” that links in situ and ex situ methods and tools. She is particularly interested in exploring

the effects of land use and climate change on chelonian conservation and how restoration ecology

may contribute to the persistence of Testudines. Photo by Ricardo Madrifian-Valderrama.

Cristian Ramirez-Gallego is a Biologist with a degree from the Universidad de Antioquia in

Medellin, Colombia, and a Master’s degree from the Universidad de Puerto Rico, Rio Piedras

campus in San Juan, Puerto Rico. He was awarded the Archie Carr Student Award — Runner-up:

Biology, by the international Sea Turtle Society in 2014, and has published over 16 scientific articles

and book chapters. His research interests are on the nesting ecology and conservation genetics

of sea turtles, illegal commerce of sea turtles and their products, with a focus on “tortoiseshell,”

as well as in strengthening the technical capacities of communities and public entities for sea

turtle management and conservation. He is a co-founder of the Fundacion Tortugas del Mar in

Colombia and an Associate Investigator of the Corporacion para el Desarrollo de la Costa Caribe —

CORPOCARIBE. Photo by Cristian Ramirez-Gallego.

Jennifer Sofia del Rio is a Biologist with a degree from the Universidad Distrital Francisco José de

Caldas in Bogota, Colombia, and is finishing another degree in Biology at the Universidad INCCA

de Colombia. She works for WWF Colombia, focusing on protected area planning and management

effectiveness assessments and the implementation of the IUCN Green List Program in Colombia.

She has experience in promoting the establishment of private protected areas, working with private

landowners in biodiversity conservation and the sustainable use of biological resources. Jennifer

also has expertise in the biology and conservation ecology of freshwater turtles and is interested in

acoustic communication, conservation strategies, and population ecology of turtles. Photo by Sindy

Martinez.

Marley T. Gémez-Rinco6n is a Biologist from the Universidad de Antioquia in Medellin, Colombia.

Her interests are focused on the population ecology, conservation, and biology of freshwater turtles.

Photo by Jennifer Del Rio.

Margarita M. Vallejo-Betancur obtained her Master’s degree in Biological Sciences from the

Universidad CES and Escuela de Ingenieria de Antioquia, Colombia, in 2018. She is interested in

animal behavior and demography, and has worked on strategies for preventing and combating illegal

wildlife trafficking as well as wildlife management and welfare. She conducted an internship in

wildlife conservation in the United Kingdom and has experience in wildlife conservation centers in

Oceania. Photo by Juan Pablo Lopera.

135 April 2022 | Volume 16 | Number 1 | e306

Official journal website:

amphibian-reptile-conservation.org

Amphibian & Reptile Conservation

16(1) [General Section]: 136-147 (e307).

Distribution range expansion of Salamandra infraimmaculata

Martens, 1885 (Caudata: Salamandridae) in Anatolia, Turkey,

with a new locality record

*Kamil Candan

Dokuz Eylul University, Faculty of Science, Department of Biology, Buca-Izmir, TURKEY

Abstract.—Salamandra infraimmaculata is one of the prominent members of the Turkish herpetofauna, which

is currently classified as Near Threatened by the IUCN. Although many studies of its morphology, ecology,

and phylogeny have been conducted in recent years, many issues concerning its taxonomic structure and

morphometry remain unresolved. In the present study, morphometric characters and color-pattern features

of the specimens of Salamandra infraimmaculata captured from ivriz, Halkapinar, Konya Province are given

in detail and compared with the data available in previous studies. In addition, the climatic niche preferences

of this species were determined using ecological niche modelling, and three WorldClim bioclimatic variables

were found to restrict the species presence: Minimum Temperature of Coldest Month (Bio6), Mean Temperature

of Wettest Quarter (Bio8), and Precipitation of Warmest Quarter (Bio18). Even though this study revealed the

potential habitat preferences of this species, clearly more detailed studies are needed to resolve the problematic

taxonomical issues.

Keywords. Amphibia, color pattern, morphometry, niche modelling, WorldClim bioclimatic variables, ecological niche

Citation: Candan K. 2022. Distribution range expansion of Salamandra infraimmaculata Martens, 1885 (Caudata: Salamandridae) in Anatolia, Turkey,

with a new locality record. Amphibian & Reptile Conservation 16(1) [General Section]: 136-147 (e307).

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.

Accepted: 6 December 2020; Published: 15 March 2022

Introduction

The genus Salamandra currently includes six recognized

species: S. algira Bedriaga, 1883; S. atra Laurenti, 1768;

S. corsica Savi, 1838: S. infraimmaculata Martens, 1885;

S. lanzai Nascetti, Andreone, Capula and Bullini, 1988:

and S. salamandra Linnaeus, 1758 (Rodriguez et al.

2017). One of them, the Near Eastern Fire Salamander S.

infraimmaculata, is amember of the Turkish herpetofauna

(Baran etal. 2012). Salamandra infraimmaculata was first

described as Salamandra maculosa vat. infraimmaculata

from Bcharré, Lebanon (Martens 1885). This species

is now known from Turkey, Syria, Lebanon, northern

Israel, northern Iraq, and western Iran and includes three

subspecies: S. i. infraimmaculata Martens, 1885, S. 7.

orientalis Wolterstorff, 1925, and S. i. semenovi Nesterov,

1917 (Joger and Steinfartz 1995; Steinfartz et al. 2000).

In Turkey, these three subspecies are found in Hatay

province (S. 7. infraimmaculata), in Adana, Mersin, and

Malatya provinces of southern Turkey (S. i. orientalis),

and in Bitlis and Erzincan provinces of eastern Turkey

(S. i. semenovi) (Joger and Steinfartz 1995; Veith et al.

Correspondence. ‘kamil.candan@deu.edu.tr

Amphib. Reptile Conserv.

1998; Steinfartz et al. 2000).

Specimens of this taxon from Turkey were previously

treated as S. salamandra (Eiselt 1966; Schmidtler

and Schmidtler 1970; Oz 1987; Oz and Arikan 1990;

Arikan et al. 1990; Baran and Oz 1994). However, the

Kemaliye (Erzincan) population was assigned to S. s.

infraimmaculata (Fachbach 1971), and the subspecies

was later elevated to the full species level based on

serum protein patterns (Gasser 1978). According to

comparisons of blood-serum proteins among the various

Salamandra populations by polyacrylamide disc gel

electrophoresis, the populations in Turkey were classified

as S. infraimmaculata (Joger and Steinfartz 1995). The

study by Bohme et al. (2013) on various attributes of

S. infraimmaculata reported that S. i. semenovi has a

typical scrolled pattern of yellow rings, semicircles, and

similar pattern elements, while the color pattern of both

nominates of form S. 7. orientalis has larger broad solid

flecks and small, yellow spots over the whole body except

for the belly. In S. 7. infraimmaculata, the yellow dots are

large and extend over the whole body, except the belly.

There are usually four yellow spots on the head, one on

each parotoid and one above each eye (Rastegar-Pouyani

March 2022 | Volume 16 | Number 1 | e307

Candan

and Fizi 2006). Previous molecular phylogenetic studies

identified six major monophyletic groups belonging to

this genus, which were separated from each other by 5

to 13 million years (Veith et al. 1998; Steinfartz et al.

2000; Weisrock et al. 2006; Vences et al. 2014; Pyron and

Wiens 2011; Rodriguez et al. 2017).

One molecular study claimed that the populations

occupying the Zagros Mountains (Iran) are genetically

close to the southeastern populations of Turkey, and it has

been suggested that some populations in southern Turkey

can be considered as a new subspecies because the type

locality of S. i. semenovi is in the other clade (Ahsani

et al. 2019). Based on additional herpetological studies

in Turkey, the known geographical distribution of S.

infraimmaculata has recently been expanded (Coskun et

al. 2013; Olgun et al. 2015; Sarikaya et al. 2017; Akman

et al. 2018; Sami and Yildiz 2018; Yildiz et al. 2019),

although the taxonomic state of the species division into

three subspecies 1s still not fully clarified (Steinfartz et al.

2000; Olgun et al. 2015; Ahsani et al. 2019).

In this study, the morphological characters of S.

infraimmaculata salamanders collected from a new

locality in Turkey are described, which may shed some

light on the taxonomy of this species. In addition, the

climatic conditions affecting the distribution of S.

infraimmaculata were determined using ecological niche

modelling.

Materials and Methods

Sampling. On 17 November 2019, one adult male (Fig.

1A) and one adult female (Fig. 1B) of S. infraimmaculata

were collected from Ivriz, Halkapinar, Konya Province

(Fig. 2). The salamanders used in this study were

found under stones during the daytime (between 1100

and 1200 h) and collected by hand after a heavy rain.

The localities where the specimens were collected are

situated near small creeks, and their geographic positions

were recorded with a GPS receiver (Garmin eTrex 30).

The sex of the captured individuals was determined

through secondary sexual characters, i1.e., males have a

prominent (swollen) cloaca (BasoZlu and Ozeti 1973).

Morphometric measurements and the color and pattern

characteristics of the specimens were recorded in the field

and the specimens were released at the capture locations.

Morphological characteristics. All morphological

measurements were recorded using a digital caliper

(Mitutoyo, Kawasaki, Japan) with an accuracy of 0.01

mm. Measurements of body parts and their ratios follow

previously published papers on salamanders (Oz 1987;

Olgun et al. 2015), and are as follows: total body length

(TBL), tip of snout to tip of tail; body length (LCP),

length from snout to anterior end of cloaca opening;

snout-vent length (SVL), tip of snout to posterior end of

cloaca opening; tail length (TL), length from posterior end

of cloaca opening to tip of tail; forelimb length (FLL);

Amphib. Reptile Conserv.

hindlimb length (HLL); distance between fore- and hind

limbs length (DFHL); head length (HL), distance from

snout to gular fold; head width (HW); parotoid length (PL);

parotoid width (PW); and distance between anterior of each

parotoid (DAP). In addition, the ratios of DFHL/SVL,

PL/HL, PW/HW, PW/PL, HW/HL, TL/SVL, HL/LCP,

FLL/SVL, TL/SVL*100, HL/LCP*100, HW/HL*100,

FLL/SVL* 100, HLL/SVL*100, and TL/TBL*100 were

calculated. Statistical analyses were carried out using

STATISTICA 6.0 (StatSoft, Inc., Tulsa, Oklahoma, USA)

to determine descriptive statistics for the measurements of

the salamanders. Morphometric measurements were then

compared to previously published data (Oz 1987; Oz and

Arikan 1992; Olgun et al. 2015).

Ecological niche modelling. A total of 56 records

were collected from the published literature and on-

going fieldwork over the course of a few years (Fig. 2;

Supplementary Table 1). The 56 records were thinned to

A4 localities using ‘spThin’ (Aiello-Lammens et al. 2015)

in R (R Core Team, 2019), and the thinning distance

was selected as 10 km. WorldClim bioclimatic variables

(Biol—19, Supplementary Table 2) were downloaded at

0.5 arcmin resolution using the raster package (Hijmans

2017) as environmental predictors. The minimum convex

polygon was used as the background extent of the study

region, and buffered by 0.5 degrees. Predictor rasters

by the background extent and the random background

points sampled were masked with values of 10,000. These

processes were handled with the sp (Pebesma and Bivand

2019) and rgeos (Bivand and Rundel 2019) packages in

R. The occurrence records were partitioned by the block

method (k = 4) using the ENMeval (Muscarella et al. 2014)

package in R, and MaxEnt was run successfully with

output evaluation results for 45 clamped models. Wallace,

which is a flexible platform for reproducible modeling of

species niches and distributions, was used for the complex

workflows above (Kass et al. 2018). Later, according to the

lowest AICc value (Supplementary Table 3), MaxEnt was

run again with the following parameters: Linear, Hinge

and Quadratic as features; regularization multiplier: 4; and

number of replicates: 30. The Area Under the Receiver

Operating Characteristic (ROC) Curves (AUC) value,

averaged over the 30 replicated runs, was considered as

an additional measure of model performance. Models

with AUC = 0.5 indicate a performance equivalent to

“random,” AUC > 0.7 indicates “useful” models, and

AUC = 0.9 indicates models with “excellent” performance

(Manel et al. 2001).

Results

Descriptive statistics of the metric measurements of

the specimens are given in Table 1. The total length of

specimens (TBL) was measured as 226.83 mm for the

male and 155.85 mm for the female. The SVL of the male

was 134.92 mm, while it was 99.25 mm for the female.

March 2022 | Volume 16 | Number 1 | e307

138

S CE-DE cS OV O01 LOT CY 8E C 0OT +s THL/TL cS 0 co 0 cO.0 cO'0 vs 0 Cc ‘TAS/THAG

& CLAGE, 00 €€ €10 810 88 °CE Cc 0O0T»TAS/TTH Leel OV El 89 0 £60 6L Cl (6 dvd

= 00 CE 00 €€ 0S 0 IL 0 OS CE C 00Ts TAS/TTA 09'S vOL cO | sv 99 Cc Md

s 00 6L 00 98 OS € S6V OS C8 Cc 00T + TH/MH Ie el OL 91 OL I OVC sOSI Cc Id

00 0€ 00 0€ 000 000 00 0€ C 0010 xdO'T/TH 90:0¢ 86 VC 97 C 8V CS CE C MH

= cO LS cI 89 gos S8L LS C9 Cc 0OT"TAS/TL 9V EC DOES: OTL 08'S 98° 1G Cc TH

A ce 0 €£0 S00 10.0 €£0 6 TAS/TIHA IL vs 60 OL 69'L 88 Ol OV c9 C THA

8 0€ 0 0€ 0 00:0 000 0€ 0 Cc dO'VTH IS Ce 6l vV v8'S 978 SE 8E Cc TIH

2 LS0 89 0 80 0 900 £90 Cc "TAS/TL 88 TE 80 €V 09'S c6 L 8V LE (6 TH

oS 6L 0 98 0 700 $00 €8 0 Cc TH/MH 09°9¢ 16 16 99 LI L6 VC SC VL ¢ TL

4 cv 0 9V 0 cO'0 £00 vv 0 Cc "Id/Md SC 66 c6 VET 98: LI CC SC 60 LIT Cc ‘TAS

os 8c 0 Ie 0 cO.0 cO.0 0€ 0 6 MAH/Md €C 6L 10 LOI 16 €l 99 6l VI £6 Cc dO'T

S €¢ 0 LS 0 CO 0 €00 gf 0 C TH/Id S8 SSI €8 97 6y SE 61 0S ve lol ¢ TAL

cS 5 2 aC iS ‘as uvdIN N $19) B1BYD 5 2) aC ‘as UvdIN N s.19)B1vYD

3 S+P S+P

= ‘SpOULOP PUL STRIIOJVI] Ul UDATS OTe SId}OVILY JO

= SUONLIADIQGE SY} SULOUT SY} JO JOLIO plepueys “q's SUONIADP psepurys “q's ‘saydures Jo Joquinu ‘Ny ‘sopdures DyDINIDUUIDA{UL DAPUDUUIDIVS JO SIDIPUI pu “SOT}eI “SIOJOVIeYO [BINSUd| *[ IIGBL

Amphib. Reptile Conserv.

Candan

Tr se

88°ZE

OS TE

0S 78

00°0€

LST

rS0

0€0

SS0

€8'0

6L TI

799

So'sI

CFS

99° LZ

Or'79

SE8E

8r LE

ST PL

60°L11

p16

pe l6l

Apnjs sty,

9¢°0

LEO

970

9L0

601

06 SI

O19

08°91

00'VC

00°CC

Oe EL

OE Le

OSE

O€ C6

O€ O€1

Cc ICI

09°CCC

BpANTTUBS.

Ig 0/crv 0

6€ 0/EE 0

6¢ 0/ST 0

19°0/09'0

C8 0/8L 0

OV €1/0V EI

00:°9/09'S

O0€ S1/08 91

09 OC/OE CT

Oc SC/OV 8T

00 8$/09 9F

OL PE/00 VE

OV CE/O9 VE

08°69/0L 9L

O€ VIT/08 OLT

OS 901/0$ cOl

OS €81/09 Z81

(a Buoy /a,vUT)

foun];

09 CLI

SIMA:

09 081

08 OSI

Avyeyy,

09 16!

RAVER,

OL V8 1

UBIUIZAT,

0OT x THL/ TL

00T»TAS/TTH

001s TAS/TTA

001 TH/MH

001+ dO'1/TH

001 TAS/TL

‘TAS /THAG

"Id/Md

MAH/Md

‘TH/Td

"TH/MH

dvd

“THA

TIH

‘TAS

dOT

Ta&L

UISAIA/VUepyY |

S1IJIBIBT)

(S107) Te Jo UNSIO, pur (7661) UeYLIY pue

ZOz (L861) ZO; ‘SOIpnys snotaoid sary} UI Sot} [edo] OYTOads WoL susUTdads IO} UDATS BSOY} YIM (ApNys stq}) sUdtUTDadS INO JO SAdIPUT PUR “SOIR “SJUDWISINSLAUT SINOU JO UOSLIedWIOD *Z aIqUL,

March 2022 | Volume 16 | Number 1 | e307

139

Amphib. Reptile Conserv.

Distribution range expansion of Salamandra infraimmaculata

re E kc ¥ fs Te

“ Le

Fig. 1. Samples of Salamandra infraimmaculata captured from the new locality:

(A) male and (B) female.

In the two specimens, the ground color of the upper

side of the head is black with two larger solid yellow

flecks (Fig. 1). In the male, these flecks are separated from

each other, but in the female, they are in contact. While

the female has small yellow spots on the side of the head

on a black background, the male specimen is spotless.

There is no pigmentation forming a faint dark dot on

the parotoid glands. The ground color of the dorsum is

blackish with a scattered pattern of large varying forms

of yellow maculation that are sometimes in contact with

each other. There are yellow spots on the side of the

body, especially in the female. Although the gular region

has maculation, the female has two obvious yellow flecks

rather than dot-shaped staining, which is more common

in males. In males, the ventral surface shows sparse

dots consisting of small yellow spots, while females

have purely black undersides. Both specimens show

medium-sized yellow flecks on the black background of

the extremities (Fig. 1). Comparative morphological data

Amphib. Reptile Conserv.

on the populations of this species in different regions of

Turkey are given in Table 2.

The average test AUC for the replicate runs was

0.863, and the standard deviation was 0.145. Among the

bioclimatic variables, Bio6 showed the highest percentage

contribution (51.9%), whereas Bio18 and Bio8 had lower

percentages of 20.7 and 12.1, respectively, and all other

variables were under 10%. The optimal habitat for this

species within the minimum convex polygon appeared

to be across the middle Taurus and Amanos mountains in

southern Anatolia (Fig. 3).

Discussion

Salamandra_ infraimmaculata is a polytypic species,

which is currently recognized as three discrete

taxonomic units: S. i. infraimmaculata Martens, 1885

(Turkish Hatay, Syria, Lebanon, and northern Israel), S.

i. orientalis Wolterstorff, 1925 (south and southeastern

March 2022 | Volume 16 | Number 1 | e307

Candan

| @ Newlocality ~~” § i, semenovi

@ Literaturedatal) —_— j. orientalis

S. i. infrimmaculata

200

Kilometers

Fig. 2. Distribution patterns of Salamandra infraimmaculata throughout southern Anatolia together

with the new locality record.

— ~~

Suitabilit

= igh : 0.88

Low : 0.00028

Fig. 3. Predicted distribution of Salamandra infraimmaculata under current climatic conditions. Warm

colors (red and yellow) show suitable habitats, whereas the blue color represents unsuitable habitats for

S. infraimmaculata.

Turkey), and S. i. semenovi Nesterov, 1917 (easternmost

Turkey, western Iran, and northern Iraq) (Joger and

Steinfartz 1995; Steinfartz et al. 2000; Bohme et al.

2013). However, the taxonomic status of its subspecies

and their distributions are still unclear (Steinfartz et al.

2000; Bohme et al. 2013).

The new locality record presented in this study

extends the known range of S. infraimmaculata by about

70 km to the northwest, as measured from the nearest

previous locality of Findikipinar1, Mersin. The new

locality (Ivriz, Halkapinar, Konya) indicates that the

distribution area of the species may extend throughout

Amphib. Reptile Conserv.

the Eastern Taurus Mountains to the western direction.

Several studies have reported data on the body sizes

of the three subspecies. Schorn and K wet (2010) reported

that S. 7. orientalis is smaller in size than the other two

subspecies (S. i. infraimmaculata and S. i. semenovi).

The maximum value for Israeli Fire Salamanders (S.

i. infraimmaculata) was given as 316 mm by Eiselt

(1958), while it was reported as 202 mm for Iraqi

specimens corresponding to S. i. semenovi (Bohme et

al. 2013). Finally, within the distribution area of the

S. i. orientalis, Bohme et al. (2013) reported the total

length of specimens from Findikpinari, Mersin, and Ilica

March 2022 | Volume 16 | Number 1 | e307

Distribution range expansion of Salamandra infraimmaculata

-150 -100 -50 0 50

Response of S. infraimmaculata to BIO6

100

Logistic output (probability of presence)

°o o o o o o o o

to i) we we = o im nm

n f=) mn o n o om

-10 0 10 20 30

Logistic output (probability of presence)

—

-50 0 50 100

Response of S. infraimmaculata to BIO8

-100

40 50 60 70 80 90

Response of S. infraimmaculata to BIO18

Fig. 4. The marginal response curves of S. infraimmaculata to (A) Minimum Temperature of Coldest Month (B106),

(B) Mean Temperature of Wettest Quarter (Bio8), and (C) Precipitation of Warmest Quarter (Biol18). The red lines

and blue shading respectively show the mean responses of the 30 replicate MaxEnt runs and the mean plus/minus one

standard deviation.

in Turkey as 200 mm and those from Kahramanmaras,

Turkey as 255 mm. In the present study, the total body

length was 226.83 mm for the male specimens. As can

be seen from the values given above, the suggestion that

adults of S. i. orientalis are smaller than the other two

forms, as Bohme et al. (2013) stated, is not correct.

According to Bohme et al. (2013), S. i. semenovi has

the typical, scrolled pattern of yellow rings, semicircles,

and similar pattern elements, while the color pattern of

both S. i. orientalis and the nominotypical form consists

of larger solid flecks. The color-pattern characteristics

alone are not enough to distinguish the Fire Salamanders

for taxonomically assigning individuals (Bohme et al.

2013). However, phylogenetic and phylogeographic

information obtained from different molecular

techniques can be combined with the knowledge of

morphology and distribution, producing a more accurate

taxonomic placement for the studied specimens.

Considering the niche modelling of S. infraimmaculata

in Syria, one study reported a positive relationship

between fitness and the Precipitation of Coldest

Quarter (Bogaerts et al. 2013). In this study, among

the climatic variables used in the ecological niche

modelling, Minimum Temperature of the Coldest

Amphib. Reptile Conserv.

142

Month (Bio6), Mean Temperature of Wettest Quarter

(Bio8), and Precipitation of Warmest Quarter (Bio18)

are the most important. Generally, in southern

Anatolia, S. infraimmaculata can survive in the low

temperatures in winter months. In other words, the

known occurrences for S. infraimmaculata have

minimum temperatures between -15 and 0 °C (Fig.

AA). Similarly, S. infraimmaculata can tolerate mean

temperatures between -8 and 0 °C in summer months

(Fig. 4B). Additionally, the species is not found in

localities with high precipitation, whereas it can adapt

to low precipitation in the summer months (Fig. 4C). In

particular, the temperature niches of S. infraimmaculata

can explain local population abundances in the south of

Anatolia (Bowler et al. 2015).

There are still many uncertainties regarding the

taxonomic structure of S. infraimmaculata, which

makes it difficult to determine the distribution limits

of the taxonomic groups within this species. In this

study, a new locality record for the species is reported

by revealing its potential habitat preferences. However,

more detailed studies are needed to fully resolve

the taxonomic uncertainties and to more thoroughly

document the distribution preferences of the species.

March 2022 | Volume 16 | Number 1 | e307

Candan

Literature Cited

Ahsani N, Kaboli M, Rastegar-Pouyani E, Karami

M. 2019. Genetic structure of Salamandra

inframaculata Martens, 1885 in Kurdistan Province,

Western Iran. Russian Journal of Herpetology 26(5):

267-271.

Aiello-Lammens ME, Boria RA, Radosavljevic A,

Vilela B, Anderson RP. 2015. spThin: an R package

for spatial thinning of species occurrence records

for use in ecological niche models. Ecography 38:

541-545.

Akman B, Yildiz MZ, Ozcan AF, Bozkurt MA, [&ci N,

Go¢men B. 2018. On the herpetofauna of the East

Anatolian Province of Bitlis (Turkey) (Amphibia;

Reptilia). Herpetozoa 31(1/2): 69-82.

Arikan H, Ozeti N, Oz M. 1990. DoZu Anadolu’dan

Bitlis = Salamandra — salamandra _ (Urodela,

Salamandridae) Populasyonunun Serum Proteinleri

Uzerene Bir Calisma. Turkish Journal of Zoology

14: 188-194.

Baran [, Oz M. 1994. Salamandra salamandra of

Anatolia. Mertensiella 4: 25-32.

Baran I, Ilgaz C, Avc1 A, Kumlutas Y, Olgun K. 2012.

Tiirkiye Amfibi ve Siiriingenleri. TUBITAK Popiler

Bilim Kitaplar1, Ankara, Turkey. 204 p.

BasoSlu M, Ozeti N. 1973. Turkiye amfibileri. Ege

Universitesi Fen Fakiiltesi Kitaplar Serisi 151:

1-155.

Bivand R, Rundel C. 2019. Rgeos: interface to

geometry engine - open source (‘GEOS’). R version

0.3-26. Available: https://CRAN.R-project.org/

package=rgeos [Accessed: 5 December 2019].

Bogaerts S, Sparreboom M, Pasmans F, Almasri A,

Beukema W, Shehab A, Amr ZS. 2013. Distribution,

ecology, and conservation of Ommatotriton

vittatus and Salamandra infraimmaculata in Syria.

Salamandra 49(2): 87-96.

Bohme W, Hartmann T, Fleck J, Schottler T. 2013.

Miscellaneous notes on Oriental Fire Salamanders

(Salamandra_ infraimmaculata Martens, 1885),

(Lissamphibia: Urodela: Salamandridae). Russian

Journal of Herpetology 20(1): 66-72.

Bowler DE, Haase P, Kroncke I, Tackenberg O, Bauer

HG, Brendel C, Brooker RW, Gerisch M, Henle Kk,

Hickler T, et al. 2015. A cross-taxon analysis of the

impact of climate change on abundance trends in

central Europe. Biological Conservation 187: 41-

50.

Coskun Y, Kaya A, Kaya C. 2013. Salamandra

infraimmaculata (Mertens, 1948) ve Neurergus

strauchii (Steindachner, 1887) (Caudata: Salaman-

dridae) i¢in GiineydoSu Anadolu Bolgesi’nden

yeni kayitlar [New records of Salamandra

infraimmaculata (Mertens, 1948) and Neurergus

strauchii (Steindachner, 1887) (Caudata:

Salamandridae) from southeast Anatolia]. Anadolu

Amphib. Reptile Conserv.

Doga Bilimleri Dergisi 4(1): 1-5.

Eiselt J. 1966. Ergebnisse zoologischer Sammelreisen

in der Turkei: Amphibia Caudata. Annalen des

Naturhistorischen Museums in Wien 69: 427-445.

Fachbach G. 1971. Zur Klaérung verwandtschaftlicher

Beziehungen bei Vertretern der Gattung Salamandra

mit Hilfe der Polyacrylamid-Disk-Elektrophorese.

II. Zeitschrift fiir Zoologische Systematik und

Evolutionsforschung 9: 181-187.

Gasser F. 1978. Le polytypisme del’ espeéce paléarctique

Salamandra salamandra (L.)(Amphibien. Urodele).

Archives de Zoologie Expérimentale et Générale 19:

585-617.

Hijmans RJ. 2017. Raster: geographic data analysis and

modeling. R package ver. 2.6-7. Available: https://

cran.r-project.org/web/packages/raster/index. html

[ Accessed: 5 December 2019].

Joger J, Steinfartz S. 1995. Protein electrophoretic

data on taxonomic problems in East Mediterranean

Salamandra (Urodela: Salamandridae). Pp. 33-36

In: Scientia Herpetologica (SEH). Editors, Llorente

GA, MontoriA, Santos X, Carretero MA. Asociacion

Herpetolégica Espafiola, Barcelona, Spain. 383 p.

Kass JM, Vilela B, Aiello-Lammens ME, Muscarella

R, Merow C, Anderson RP. 2018. Wallace: a

flexible platform for reproducible modeling of

species niches and distributions built for community

expansion. Methods in Ecology and Evolution 9:

1,151-1,156.

Manel S, Williams HC, Ormerod SJ. 2001. Evaluating

presence-absence models in ecology: the need to

account for prevalence. Journal of Applied Ecology

38: 921-931.

Martens V. 1885. Uber Vorkommen und Zeichnungs-

Varietaten von Salamandra_ maculosa, aus

Veranlassung eines Exemplares vom Berge Pelion in

Thessalien, welches das zoologische Museum dahier

von Hrn. Th. v. Heldreich in Athen erhalten hat.

Sitzungsberichte der Gesellschaft Naturforschender

Freunde zu Berlin 1884: 193-195.

Muscarella R, Galante PJ, Soley-Guardia M, Boria RA,

Kass JM, Uriarte M, Anderson RP. 2014. ENMeval:

an R package for conducting spatially independent

evaluations and estimating optimal model

complexity for MaxEnt ecological niche models.

Methods in Ecology and Evolution 5: 1,198—1,205.

Olgun K, Avc1 A, Bozkurt E, Uztim N, Tural M, Olgun

MF. 2015. Range extensions of two salamanders

(Neurergus strauchii (Steindachner, 1887) and

Salamandra_ infraimmaculata Martens, 1885)

(Caudata: Salamandridae) from Anatolia, Turkey.

Russian Journal of Herpetology 22: 289-296.

Oz M. 1987. Anadolu’daki Salamandra_ salaman-

dra’nin Taksonomi, Biyoloji ve Dagilisi Uzerine

Arastirmalar. Turkish Journal of Zoology 11(3):

136-154.

Oz M, Arikan H. 1990. Bitlis cevresindeki Salamandra

March 2022 | Volume 16 | Number 1 | e307

Distribution range expansion of Salamandra infraimmaculata

salamandra (Urodela, Salamandridae) populasyonu

uzerinde taksonomik arastirmalar. Turkish Journal

of Zoology 14: 195-199.

Pebesma EJ, Bivand RS. 2019. Classes and methods

for spatial data in R. R News 5(2): 9-13.

Pyron RA, Wiens JJ. 2011. A large-scale phylogeny

of Amphibia including over 2,800 species, and a

revised classification of extant frogs, salamanders,

and caecilians. Molecular Phylogenetics and

Evolution 61: 543-583.

R Core Team. 2019. R: a language and environment for

statistical computing. R Foundation for Statistical

Computing, Vienna, Austria. Available: http://

www.R-project.org/ [Accessed: 5 December 2019].

Rastegar-Pouyani N, Fizi H. 2006. On a collection

of the Near East Fire Salamander, Salamandra

infraimmaculata semenovi (Salamandridae) from

Kurdistan province, western Iran. Zoology in the

Middle East 37: 115-118.

Rodriguez A, Burgon JD, Lyra M, Irisarri I, Baurain D,

Blaustein L, Gog¢men B, Kiinzel S, Mable BK, Nolte

AW, et al. 2017. Inferring the shallow phylogeny

of true salamanders (Salamandra) by multiple

phylogenomic approaches. Molecular Phylogenetics

and Evolution 115: 16—26.

Sami E, Yildiz MZ. 2018. Amphibians of Ad1yaman/

Turkey Province. Biological Diversity and

Conservation 11(1): 1-12.

Sarikaya B, Yildiz MZ, SezenG. 2017. The herpetofauna

of Adana Province (Turkey). Kommagene Biyoloji

Dergisi 1(1): 1-12.

Schmidtler JJ, Schmidtler JF. 1970. Morphologie,

Biologie, und Verwandtschafts beziehungen von

Neurergus strauchii aus der Turkei. Senckenbergiana

Biologica 51: 41-53.

Schorn S, Kwet A. 2010. Feuersalamander.

NTVVerlag, Munster, Germany. 141 p.

Steinfartz S, Veith M, Tautz D. 2000. Mitochondrial

sequence analysis of Salamandra taxa suggests

old splits of major lineages and _ postglacial

recolonizations of central Europe from distinct

source populations of Salamandra_ salamandra.

Molecular Ecology 9: 397-410.

Veith M, Steinfartz S, Zardoya R, Seitz A, Meyer A.

1998. A molecular phylogeny of ‘true’ salamanders

(family Salamandridae) and the evolution of

terrestriality of reproductive modes. Journal of

Zoological Systematics and Evolutionary Research

36: 7-16.

Vences M, Sanchez E, Hauswaldt JS, Eikelmann

D, Rodriguez A, Carranza S, Donaire D, Gehara

M, Helfer V, Loétters S, et al. 2014. Nuclear and

mitochondrial multilocus phylogeny and survey

of alkaloid content in true salamanders of the

genus Salamandra (Salamandridae). Molecular

Phylogenetics and Evolution 73: 208-216.

Weisrock DW, Papenfuss TJ, Macey JR, Litvinchuk SN,

Polymeni R, Ugurtas IH, Zhao E, Jowkar H, Larson

A. 2006. A molecular assessment of phylogenetic

relationships and lineage accumulation rates within

the family Salamandridae (Amphibia, Caudata).

Molecular Phylogenetics and Evolution 41: 368—

383.

Yildiz MZ, Sarikaya B, Bozkurt MA. 2019. The

herpetofauna of the Province of Hatay (East

Mediterranean Turkey). Biological Diversity and

Conservation 12(2): 197-205.

Kamil Candan is currently working as a researcher at Dokuz Eyltil University, Department

of Biology in Buca-Izmir, Turkey. He obtained his M.Sc. and Ph.D. degrees for studies on

Amphib. Reptile Conserv.

the molecular phylogenetics of species belonging to the genera Anatololacerta and Darevskia

(Lacertidae), respectively, and has continued to publish additional articles on Turkey’s

herpetofaunal biodiversity. His research focuses on the molecular systematics, ecology, and

distribution of amphibians and reptiles.

March 2022 | Volume 16 | Number 1 | e307

Candan

Supplementary Table 1. Presence data of the Salamandra infraimmaculata localities used in the model.

Locality

Ivriz, Halkapinar, Konya

Maden Village, Ulukisla, Nigde

Camltyayla, Mersin

Findikpinar, Mersin

Gozne, Mersin

Omerli Village, Pozanti, Adana

Kamusli, Pozanti, Adana

Akcatekir, Pozanti, Adana

Belemedik, Pozanti, Adana

Saimbeyli, Adana

Feke, Adana

Kozan, Adana

Boztahta, Kozan, Adana

Culluusag1, Kozan, Adana

Camdere, Kozan, Adana

Baraj, Kozan, Adana

Yamanli, Tufanbeyli, Adana

Camlibel, Aladag, Adana

Belenkoy, Adana

Daripinari, Adana

Bahc¢e, Osmaniye

Kozakh Village, Iskenderun, Hatay

Iskenderun, Hatay

Bekbele Plateau, Iskenderun, Hatay

Demen Plateau, Iskenderun, Hatay

Cardak Plateau, Hassa, Hatay

Egribucak, Hassa, Hatay

Kapili yolu, Dértyol, Hatay

Harbiye, Hatay

Kozkalesi, Altinézt, Hatay

Yenikoy, Samandag, Hatay

Arkitca, Kirikhan, Hatay

Altinboga Village, Andirin, Kahramanmaras

Ilica Village, Kahramanmaras

Sugozu, Besni, Adryaman

Yasmurlu village, Gerger, Adryaman

Tut, Adiyaman

Kaslica, Tut, Adiyaman

Meydank6oy, Golbas1, Adryaman

Harmanli, Gélbas1, Adryaman

Salihli Village, Kemaliye, Erzincan

Kemaliye, Erzincan

Yuva Village, Kemaliye Erzincan

Amphib. Reptile Conserv.

Latitude

37.430556

37.448889

37.183016

36.917778

37.005833

37.528056

37.526944

1D)

31329722

37.365833

38.000833

37.810833

37.444722

37.635556

37.680278

S7653333

37512222

38.182307

37.474429

37.865833

37.141883

SP 2VI222

36.479444

36.585556

36.612550

36.635556

36.838611

36.750833

36.864444

36.124722

36.099980

36.222778

36.509167

37.626096

37.839878

37.669669

37.968325

37.808611

37.803813

37.810277

37.843888

39330269

39.266111

39.247924

145

Longitude

34.181111

34.630278

34.591682

34.372223

34.559722

34.8575

34.851111

34.955556

34.786389

34.921944

36.090556

35.919167

35.807778

36.011388

35.864167

35.816389

35.863889

36.242130

35.068174

35.855277

34.725625

36.573055

36.122778

36.218333

36.229809

36.244167

36.443611

36.454166

36.399167

36.131944

36.189077

35.986389

36.294999

36.340641

36.870419

37.786397

39.029137

37M

1979159

37.644999

37.758056

38.502082

38.4975

38.509211

March 2022 | Volume 16 | Number 1 | e307

Reference

This study

Baran and Oz (1994)

Olgun et al. (2015)

Sarikaya et al. (2017)

Baran and Oz (1994)

Sarikaya et al. (2017)

Bohme et al (2013)

Ozeti and Yilmaz (1994)

Baran and Oz (1994)

Olgun et al. (2015)

Yildiz et al. (2019)

Olgun et al. (2015)

Yildiz et al. (2019)

Olgun et al. (2015)

Sami and Yildiz (2018)

Baran and Oz (1994)

Olgun et al. (2015)

Distribution range expansion of Salamandra infraimmaculata

Supplementary Table 1 Continued. Presence data of the Salamandra infraimmaculata localities used in the model.

Locality Latitude Longitude Reference

Aslantepe, Malatya 38.381928 38.361328 Baran and Oz (1994)

Gunduizbey Village, Malatya 38.278889 38.269444 Olgun et al. (2015)

Eskihalfeti, Sanliurfa ST ASOIDZ 37.871666 Olgun et al. (2015)

Tagar Stream, Cemisgezek, Tuncelli 39.064444 38.905278 Olgun et al. (2015)

Sttlice, Tunceli 39.036944 39.165833 Olgun et al. (2015)

Gelinodalar1, Pilimir, Tunceli 39.470112 39906898 Olgun et al. (2015)

Sosuksu Village, Ergani, Diyarbakir 38.399089 39.656633 Olgun et al. (2015)

Sagirkaya, Hizan, Bitlis 37.991108 42.569625 Akman et al. (2018)

Alatepe Village, Ilica, Bingél 39.059281 40.756134 Cicek et al. (2017)

Deringay Village, Karliova Bingol 39.13 2289 40.824913 Cicek et al. (2017)

Deliktas, Bitlis 38.347686 42.041075 Akman et al. (2018)

K6sepinar1, Osmaniye 37.595694 36.155118 Unpub. data (Serkan Gil)

Supplementary Table 2. Definitions of the 19 WorldClim bioclimatic variables.

Variable Definition

BIOI Annual Mean Temperature

BIO2 Mean Diurnal Range (Mean of monthly (max temp - min temp))

BIO3 Isothermality (BIO2/BIO7) (<100)

BIO4 Temperature Seasonality (standard deviation x100)

BIOS Max Temperature of Warmest Month

BIO6 Min Temperature of Coldest Month

BIO7 Temperature Annual Range (BIOS-BIO6)

BIO8 Mean Temperature of Wettest Quarter

BIO9 Mean Temperature of Driest Quarter

BIOI10 Mean Temperature of Warmest Quarter

BIOI1 Mean Temperature of Coldest Quarter

BIO12 Annual Precipitation

BIOI3 Precipitation of Wettest Month

BIO14 Precipitation of Driest Month

BIOI5 Precipitation Seasonality (Coefficient of Variation)

BIO16 Precipitation of Wettest Quarter

BIOI7 Precipitation of Driest Quarter

BIO18 Precipitation of Warmest Quarter

BIOI9 Precipitation of Coldest Quarter

Amphib. Reptile Conserv. 146 March 2022 | Volume 16 | Number 1 | e307

Candan

Supplementary Table 3. Performance of the 48 models created during the evaluation process for Salamandra infraimmaculata. The

gray-shaded entry shows the settings chosen for the model. FC: Feature Classes, RM: Regularization Multiplier, AlCc: corrected

Akaike Information Criterion.

FC RM AICe FC RM AICe FC RM AICe

if 0.5 1091.86 L a 1094.35 it 3.5 1095.11

LQ 0.5 1076.99 LQ 2 1053.38 LO 3.5 1055.45

H 0.5 NA H 2 1078.45 H 3.5 1065.6

LOH 0.5 NA LOH 2 1045.37 LOH 3.5 1045.81

LOHP 0.5 NA LOHP 2 1042.53 LOHP 3.5 1051.03

id 1 1087.92 if 2.5 1090.27 L 4 1096.62

LO 1 1059.28 LO 2.5 1053.95 LQ 4 1056.35

H 1 1383.18 H 2.5 1067.06 H 4 1055.19

LOH 1 1289.72 LOH 2.5 1049.89 LOH 4 1040.85

LOHP 1 1332.03 LOHP 2.5 1049.21 LOHP 4 1065.72

L 1.5 1088.85 L 3 1091.14 L 4.5 1101.69

LO 1.5 1060.73 LO 3 1054.65 LQ 4.5 1057.37

H 1.5 1087.01 H 3 1061.92 H 4.5 1058.83

LOH 1.5 1057.83 LOH 3 1051.48 LOH 4.5 1042.09

LOHP 1.5 1069.73 LOHP 3 1048.21 LOHP 4.5 1051.51

Amphib. Reptile Conserv. 147 March 2022 | Volume 16 | Number 1 | e307

Introductory page. Xenosaurus mendozai Nieto-Montes de Oca, Garcia-Vazquez, Zufiga-Vega, and Schmidt-

Ballardo, 2013. The Granular-Scaled Lizard occurs in the states of Querétaro, from where it was first described,

and Hidalgo. The species was dedicated to Fernando Mendoza Quiyano, a Mexican herpetologist who contributed

substantially to the herpetofauna of the states in the central region of Mexico. In this study, and according to Wilson

et al. (2013a), we determined its EVS as 16, placing it in the high vulnerability category. According to IUCN, its

conservation status 1s unknown, and this species is not listed by SEMARNAT. This individual was found in the

municipality of Jacala de Ledezma, Hidalgo, near the type locality in the state of Querétaro. Photo by Christian

Berriozabal-Islas.

Amphib. Reptile Conserv. 148 April 2022 | Volume 16 | Number 1 | e308

Amphibian & Reptile Conservation

16(1) [General Section]: 148-192 (e308).

Official journal website:

amphibian-reptile-conservation.org

The herpetofauna of Queretaro, Mexico: composition,

distribution, and conservation status

'Raciel Cruz-Elizalde, 7Aurelio Ramirez-Bautista, ‘Rubén Pineda-Lopez, *Vicente Mata-Silva,

‘Dominic L. DeSantis, °Eli Garcia-Padilla, "Jerry D. Johnson, *Arturo Rocha, ®Lydia Allison Fucsko,

and ‘Larry David Wilson

‘Laboratorio de Zoologia, Facultad de Ciencias Naturales, Universidad Autonoma de Querétaro, Avenida de las Ciencias S/N, Santa Fe Juriquilla,

C. P. 76230, Querétaro, Querétaro, MEXICO *Laboratorio de Ecologia de Poblaciones, Centro de Investigaciones Bioldgicas, Instituto de Ciencias

Basicas e Ingenieria, Universidad Autonoma del Estado de Hidalgo, Km 4.5 Carretera Pachuca-Tulancingo, 42184 Mineral de La Reforma,

Hidalgo, MEXICO *Department of Biological Sciences, The University of Texas at El Paso, El Paso, Texas 79968-0500, USA *Department of

Biological and Environmental Sciences, Georgia College and State University, Milledgeville, Georgia 31061, USA *Oaxaca de Juarez, Oaxaca

68023, MEXICO °Department of Humanities and Social Sciences, Swinburne University of Technology, Melbourne, Victoria, AUSTRALIA ‘Centro

Zamorano de Biodiversidad, Escuela Agricola Panamericana Zamorano, Departamento de Francisco Morazan, HONDURAS and 1350 Pelican

Court, Homestead, Florida 33035-1031, USA

Abstract.—The herpetofauna of the state of Querétaro, Mexico, consists of 129 species, including 27 anurans,

seven caudates, 92 squamates, and three turtles. Regarding the distribution of the herpetofaunal species

among the three recognized physiographic regions in the state, the total number of species ranges from 43 in

the Transmexican Volcanic Belt to 102 in the Sierra Madre Oriental. The individual species inhabit from one to

three regions (X = 1.6). The majority (78.3%) of the native herpetofauna of Queretaro is found in one or two of

the three regions, which is of conservation significance. The majority of the remaining single-region species

inhabit the Sierra Madre Oriental (54), followed by 15 in the Central Plateau and eight in the Transmexican

Volcanic Belt. The Coefficient of Biogeographic Resemblance (CBR) indicates that the Sierra Madre Oriental

and the Central Plateau share the largest number of species (45) due to their adjacent positions, relatively large

areas, and because they contain the first and second largest numbers of species. A similarity dendrogram

based on the Unweighted Pair Group Method with Arithmetic Averages (UPGMA) demonstrates that the Central

Plateau and the Transmexican Volcanic Belt share the highest level of herpetofaunal resemblance (0.60). Within

the distributional categories, the largest numbers of species are the country endemics (67 of 129), followed

by the non-endemics (60) and the non-natives (2). The principal environmental threats to the herpetofauna

of Querétaro are the increasing and unregulated clearing of forests for farming and raising livestock, road

construction, the ever-increasing pollution of bodies of water, and the cultural perceptions of various

herpetofauna. The conservation status of the native species was evaluated by employing the SEMARNAT (NOM-

059), IUCN, and EVS systems, of which the EVS was the most useful. Using the two Relative Herpetofaunal

Priority (RHP) methods to designate the rank order significance of the physiographic regions, the highest

ranks were obtained for the Sierra Madre Oriental. In considering the features of the three protected areas in

Queretaro, we determined that two are located in the Transmexican Volcanic Belt, which is the least important

region from a conservation perspective. We also determined that only 79 of the 127 native species recorded

from Queretaro are known to occur in any of the three protected areas. Finally, we provide a set of conclusions

and recommendations in an effort to ensure the future protection of the herpetofauna of Queretaro.

Keywords. Amphibia, Anurans, caudates, physiographic regions, protected areas, protection recommendations, Rep-

tilia, squamates, turtles

Resumen.—La herpetofauna de Queretaro, Mexico, consta de 129 especies, incluyendo 27 anuros, siete

caudados, 92 escamosos y tres tortugas. Documentamos la distribucion de las especies de herpetofauna

entre las tres regiones fisiograficas que reconocemos. El numero total de especies varia de 43 en la Faja

Volcanica Transmexicana a 102 en la Sierra Madre Oriental. Las especies individuales habitan de una a

tres regiones (xX = 1,6). Una proporcion del 78.3% de la herpetofauna nativa de Querétaro se encuentra en

una o dos de las tres regiones, lo cual es de gran importancia para la conservacion. El mayor numero de

especies de una sola region habita en la Sierra Madre Oriental (54), seguido de 15 en la Meseta Central y

ocho en la Faja Volcanica Transmexicana. Un coeficiente de semejanza biogeografica (CBR) indica que la

Sierra Madre Oriental y la Meseta Central comparten el mayor numero de especies (45), debido a su ubicacion

Correspondence. cruzelizale@gmail.com (RCE), ramibautistaa@gmail.com (ARB), rpineda62@hotmail.com (RPL), vmata@utep.edu

(VMS), dominic.desantis@gcsu.edu (DLD), eligarcia_1S@hotmail.com (EGP), jjohnson@utep.edu (JDJ), lvdiafucsko@gmail.com (LAF),

bufodoc@aol.com (LDW)

Amphib. Reptile Conserv. 149 April 2022 | Volume 16 | Number 1 | e308

The herpetofauna of Queretaro, Mexico

adyacente, su area relativamente grande y su albergue del primer y segundo mayor numero de especies. Un

dendrograma de similitud basado en el metodo de grupos de pares no ponderados con promedios aritméticos

(UPGMA) demuestra la Meseta Central y la Faja Volcanica Transmexicana comparten la mayor semejanza de

herpetofauna (nivel 0.60). Con referencia a las categorias de distribucion, la mayor cantidad de especies es la

de las endémicas del pais (67 de 129), seguidas de las no endemicas (60) y las no nativas (2). Las principales

amenazas ambientales para la herpetofauna de Queretaro son la creciente y desmedida tala de bosques para la

agricultura y la ganaderia, la construccion de caminos, la constante y creciente contaminacion de los cuerpos

de agua, y la percepcion cultural de los miembros de la herpetofauna. Evaluamos el estado de conservacion

de las especies nativas empleando los sistemas SEMARNAT (NOM-059), UICN y EVS, de los cuales el sistema

EVS fue el mas util. También utilizamos los dos métodos de Prioridad relativa de la herpetofauna (RHP) para

designar la importancia del orden de clasificacion de las regiones fisiograficas y determinamos los valores mas

altos para la region de la Sierra Madre Oriental. Examinamos las caracteristicas de las tres areas protegidas en

Queretaro y determinamos que dos de las tres estan ubicadas en la Faja Volcanica Transmexicana, que es la

region menos importante desde una perspectiva de conservacion. Tambien determinamos que solo 79 de las

127 especies nativas registradas en Queretaro, se registran en total de las tres areas protegidas. Finalmente,

emitimos un conjunto de conclusiones y recomendaciones para la futura proteccion de la herpetofauna de

Querétaro.

Palabras Claves. Anfibios, anuros, areas protegidas, caudados, escamosos, regiones fisiograficas, reptiles, recomen-

daciones de proteccion, tortugas

Citation: Cruz-Elizalde R, Ramirez-Bautista A, Pineda-L6pez R, Mata-Silva V, DeSantis DL, Garcia-Padilla E, Johnson JD, Rocha A, Fucsko LA,

Wilson LD. 2022. The herpetofauna of Querétaro, Mexico: composition, distribution, and conservation status. Amphibian & Reptile Conservation 16(1)

[General Section]: 148-192 (e308).

[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.

Accepted: 30 January 2022; Published: 18 April 2022

“It was an event that changed the course of natural

history—wiping out three-quarters of all species,

including anything on land larger than the size of a

domestic dog. It ended the 175-million-year reign of the

dinosaurs. Life would have to rebuild... For 66 million

years since then, nature has been at work reconstructing

the living world, recreating and redefining a new diversity

of species. And one of the products of this rebooting of

life was humanity.”

David Attenborough (2020)

Introduction

Querétaro, a relatively small state in north-central

Mexico, is positioned in a northwest to southeast

direction at the intersection of three major physiographic

regions: the Sierra Madre Oriental in the northeast, the

Central Plateau in the middle, and the Transmexican

Volcanic Belt in the southwest (Fig. 1). To the north,

Querétaro is bounded by San Luis Potosi, to the east

by Hidalgo, to the south by México and Michoacan,

and to the west by Guanajuato. The area of Querétaro

is 11,699 km’, which ranks 27" in size among the 32

federal entities in Mexico (http://wikipedia.org; accessed

22 July 2019); only Colima, Aguascalientes, Morelos,

Tlaxcala, and Ciudad de México are smaller. Querétaro’s

area represents only 0.6% of the country, while the

human population of the state in 2015 was reported as

Amphib. Reptile Conserv.

2,038,372, which ranks 22"! (16.0%) in the country. The

population density is indicated as 170/km?, or 7" in the

country (http://wikipedia.org; accessed 22 July 2019),

which is 2.8 times the average density for Mexico. The

state of Querétaro, therefore, lies within the most densely

populated region of Mexico (1.e., the area surrounding

the Mexican metropolitan area), which includes the

states of México, Morelos, Tlaxcala, Aguascalientes,

Guanajuato, and Puebla. Only Aguascalientes is slightly

removed from the other states in this heavily-populated

region. Although a significant amount of environmental

deterioration 1s expected to occur in Querétaro, the state

still supports “a diversity of undisturbed environments...

such as cloud forest, pine forest, oak [forest], and tropical

deciduous forest” (Cruz-Elizalde et al. 2019).

The highest elevation in the state 1s 3,360 m (http://

wikipedia.org; accessed 22 July 2019) on Cerro el

Zamorano, along the border with Guanajuato in the

Central Plateau, and a communications facility is present

on this peak (http://googlemaps.com; accessed 22 July

2019). Elevations over 3,000 m also occur in the two

other physiographic regions of the state (http://wikipedia.

org; accessed 22 July 2019).

Since Querétaro encompasses portions of three major

physiographic regions in Mexico as described above,

it could be expected to have a herpetofauna somewhat

comparable to those of Puebla and/or Hidalgo. However,

Querétaro is significantly smaller than either Puebla

(34,306 km’; Woolrich-Pifia et al. 2017) or Hidalgo

April 2022 | Volume 16 | Number 1 | e308

Cruz-Elizalde et al.

100°0'0"W

22°0'0"N

San Luis Potosi

21°0'0"N

Hidalgo

20°0'0"N

100°0'0"W

99°0'0"-=W

Physiographic regions

Central Plateu (CP)

( Transmexican Volcanic Belt (TVB)

Sierra Madre Oriental (SMO)

Fig. 1. Physiographic regions in the state of Querétaro, Mexico, and location of the state of Querétaro in Mexico. The map 1s based

on INEGI (2000).

(20,813 km’; Ramirez-Bautista et al. 2020). Furthermore,

Hidalgo contains portions of four physiographic regions,

including the Gulf coastal lowlands, and Puebla contains

six regions, including the Gulf coastal lowlands and two

valley regions. Accordingly, it is more useful to compare

the herpetofaunas recorded in the same physiographic

regions in Puebla and Hidalgo, which also are represented

in Querétaro (see below).

Materials and Methods

Our Taxonomic Position

In this study we follow the taxonomic position regarding

the subspecies concept that was described 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; Nevarez-de los Reyes

et al. 2016; Cruz-Saenz et al. 2017; Gonzalez-Sanchez

et al. 2017; Woolrich-Pifia et al. 2017; Lazcano et al.

2019; Ramirez-Bautista et al. 2020). Johnson et al.

(2015a) can be consulted for a detailed statement on

this position.

Amphib. Reptile Conserv.

Construction of the Species List

We made some corrections to the recent species list

published for the state of Querétaro, as noted by Cruz-

Elizalde et al. (2019). That list consisted of 138 species,

including 34 amphibians and 104 reptiles. In the interim,

however, questions arose regarding the presence of 17

species in the state, whose status was reevaluated when

preparing this paper. We discuss the status of these

species below.

System for Determining Distributional Status

We used the system developed by Alvarado-Diaz et al.

(2013) for the herpetofauna of Michoacan to determine

the distributional status of members of the herpetofauna

of Querétaro. Subsequently, various other studies (Mata-

Silva et al. 2015; Johnson et al. 2015a; Teran-Juarez et al.

2016; Woolrich-Pifia et al. 2016; Nevarez-de los Reyes et

al. 2016; Cruz-Sanchez et al. 2017; Gonzalez-Sanchez et

al. 2017; Woolrich-Pifia et al. 2017; Lazcano et al. 2019;

Ramirez-Bautista et al. 2020) have used this system,

which consists of the following four categories: SE =

April 2022 | Volume 16 | Number 1 | e308

The herpetofauna of Queretaro, Mexico

No. 1. Jncilius occidentalis (Camerano, 1879). The Pine

Toad occurs in the mountains of Durango, Jalisco, Nayarit,

Aguascalientes, Querétaro, Hidalgo, and Veracruz, southward

through the highlands west of the Isthmus of Tehuantepec,

Mexico. This individual was found in Huimilpan, Querétaro.

Wilson et al. (2013b) determined its EVS as 11, placing it in

the medium vulnerability category. Its conservation status was

assessed as Least Concern by the IUCN, but this species is not

listed by SEMARNAT. Photo by Raciel Cruz-Elizalde.

No. 3. Eleutherodactylus verrucipes (Cope, 1885). The Big-

eared Chirping Frog is known from moderate elevations (200-

1,300 m asl) in pine-oak woodland and cloud forest located

in southeastern San Luis Potosi, Querétaro, Guanajuato, and

northwestern Hidalgo, Mexico (Frost 2021). Wilson et al.

(2013b) determined its EVS as 16, placing it in the middle

portion of the high vulnerabilty category. Its conservation

status has been assessed as Vulnerable by the IUCN, and in

the Special Protection (Pr) category by SEMARNAT. Photo by

Raciel Cruz-Elizalde.

Amphib. Reptile Conserv.

“= ORS? is eae pate ed — ee, Py

No. 2. Rhinella horribilis (Wiegmann, 1833). The Cane Toad,

an introduced species, occurs in most regions of Mexico, and

has a great effect on the native fauna. This individual was

found in Jacalilla, Querétaro. Wilson et al. (2013a) determined

its EVS as 3, placing it in the low vulnerability category. Its

conservation status has been assessed as Least Concern by the

IUCN, but this species is not listed by SEMARNAT. Photo by

Raciel Cruz-Elizalde.

No. 4. Dryophytes arenicolor (Cope, 1866). The Canyon

Treefrog occurs in the mountains and plateau regions of the

United States (southern Utah and southern Colorado southward

through eastern Arizona, western and northern New Mexico,

as well as in Nevada to about Las Vegas, and the Trans-Pecos

region of Texas); and southward in Mexico to Michoacan,

Colima, México, Guerrero, Hidalgo, Querétaro, and Oaxaca.

This individual was found in Huimilpan, Querétaro. Wilson

et al. (2013b) determined its EVS as 7, placing it in the low

vulnerability category. Its conservation status has been assessed

as Least Concern by the IUCN, but this species is not listed by

SEMARNAT. Photo by Raciel Cruz-Elizalde.

April 2022 | Volume 16 | Number 1 | e308

Cruz-Elizalde et al.

endemic to Querétaro; CE = endemic to Mexico; NE =

not endemic to Mexico; NN = non-native in Mexico.

Systems for Determining Conservation Status

To assess the conservation status of the herpetofauna

of Querétaro, we employed the three systems (e.,

SEMARNAT, IUCN, and EVS) 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), Nevarez-de los Reyes et al. (2016), Cruz-

Sanchez et al. (2017), Gonzalez-Sanchez et al. (2017),

Woolrich-Pifia et al. (2017), Lazcano et al. (2019), and

Ramirez-Bautista et al. (2020). Detailed descriptions of

these three systems have appeared in earlier papers in

this series (listed below) and are not 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) that appeared in Amphibian &

Reptile Conservation, in an issue that included five related

papers and was designated as the Special Mexico Issue.

The basic format for entries in the MCS was established

in that paper, 1.e., 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 resumed with two studies on

the herpetofauna of Oaxaca (Mata-Silva et al. 2015) and

Chiapas (Johnson et al. 2015a). The following year three

entries in the MCS were published on Tamaulipas (Teran-

Juarez et al. 2016), Nayarit (Woolrich-Pifia et al. 2016),

and Nuevo Leon (Nevarez-de los Reyes et al. 2016). In

2017, three more entries in this series were published on

Jalisco (Cruz-Saenz et al. 2017), the Mexican Yucatan

Peninsula (Gonzalez-Sanchez et al. 2017), and Puebla

(Woolrich-Pifia et al. 2017). More recently, entries were

published on Coahuila (Lazcano et al. 2019), Hidalgo

(Ramirez-Bautista et al. 2020), and most recently on

Veracruz (Torres-Hernandez et al. 2021). Thus, this study

on the herpetofauna of Querétaro is the 13" entry in the

MCS series.

Physiography and Climate

Physiographic Regions

Sierra Madre Oriental (SMO). This region (Figs.

2-7) is located parallel to the Gulf coastal region of

Mexico, and it 1s connected to the Central Plateau and

the Transmexican Volcanic Belt. The SMO has been

assigned to the Neotropical Realm and covers 2.84%

of the area of the country (Morrone 2001; CONABIO

2008). This province is composed mostly of sedimentary

and metamorphic rocks from the Cretaceous and Jurassic

periods, which makes this province a complex area

Amphib. Reptile Conserv.

from a geological perspective (CONABIO 2008). The

SMO encompasses parts of Coahuila, Nuevo Leon,

Tamaulipas, San Luis Potosi, Hidalgo, Puebla, Querétaro,

Tlaxcala, and Veracruz (CONABIO 2008). In the state of

Querétaro, this province extends into the municipalities

of Pinal de Amoles, Arroyo Seco, Jalpan de Serra, Landa

de Matamoros, and the region north of Cadereyta de

Montes and San Joaquin (Figs. 2-3; CONABIO 2008).

The mean annual precipitation is 731 mm, with a greater

intensity of rainfall in September, and the mean annual

temperature is 23.5 °C, with intervals of 10.6—33.5 °C

(Luna Soria and Suzan Azpiri 2016). The climate is

classified as semi-warm subhumid, and the predominant

types of vegetation are tropical deciduous forest, oak

forest, pine-oak, and portions of cloud forest (Bayona

Celis 2016).

Central Plateau (CP). This region (Figs. 8—9) lies within

the more inclusive Nearctic Region (Morrone 2001;

CONABIO 2008), extends through the central area of

Mexico at elevations from 1,700 to 4,000 m asl, and

is located between the Sierra Madre Occidental and

Sierra Madre Oriental. Portions of the CP fall within

the boundaries of Chihuahua, Coahuila, Durango,

Guanajuato, Hidalgo, Jalisco, Michoacan, Puebla,

Querétaro, San Luis Potosi, Tlaxcala, and Zacatecas.

In the state of Querétaro, this province extends into the

municipalities of Pefiamiller, Toliman, San Joaquin,

Cadereyta de Montes, Colon, Ezequiel Montes, El

Marqués, Querétaro, Tequisquiapan, Pedro Escobedo,

and San Juan del Rio (CONABIO 2008). The climate

is temperate semi-dry steppe, with rainfall occurring in

summer; the average annual precipitation is 460 mm;

the average annual temperature is 17.8 °C; and the

temperature ranges from 4.5 to 30 °C (Luna Soria and

Suzan Azpiri 2016). The predominant types of vegetation

are crasicaule scrub, rosetophilous scrub, microphilous

desert scrub, and gallery forest, but these regions

contain extensive areas devoted to agriculture, as well as

grasslands (Bayona Celis 2016).

Transmexican Volcanic Belt (TVB). The TVB lies in the

Neotropical Region (Morrone 2001; CONABIO 2008)

and forms a volcanic arc located in central Mexico.

The TVB is oriented from east to west, from the state

of Veracruz (Gulf of Mexico) to Nayarit (Pacific

Ocean; Ferrusquia-Villafranca 2007; CONABIO

2008). In the state of Querétaro, this province

extends into the municipalities of Amealco de Bonfil,

Corregidora, and Huimilpan (CONABIO 2008). The

predominant climate is temperate subhumid, with rain

occurring in summer, and with its greatest intensity

from May to October; the mean annual rainfall is 861

mm, and the mean annual temperature is 14.4 °C, with

intervals from 3.8 to 24.8 °C (Luna Soria and Suzan

Azpiri 2016). The predominant vegetation types are

oak and pine forests (Figs. 5—6), in addition to such

April 2022 | Volume 16 | Number 1 | e308

The herpetofauna of Queretaro, Mexico

poe

Fig. 2. View of the Sierra Madre Oriental at the Mirador Cuatro Fig. 3. Sierra Madre Oriental, in the Sierra Gorda Biosphere

Palos. Photo by Erick Daniel Velasco Esquivel. Reserve, Jalpan de Serra, Querétaro. Photo by Erick Daniel

Velasco Esquivel.

: eke 5s 2

Fig. 4. Xerophilous scrub in mountains of the Central Plateau, Fig. 5. Pine-oak forest, Toliman, Querétaro. Photo by Erick

locality of Pefiamiller. Photo by Erick Daniel Velasco Esquivel.

Fig. 6. Oak forest, Mirador Cuatro Palos, Querétaro. Photo by Fig. 7. Deciduous forest, Las Adjuntas, Arroyo Seco, Querétaro.

Erick Daniel Velasco Esquivel. Photo by Erick Daniel Velasco Esquivel.

53 yaw = “4 ~ é

Fig. 8. Juniperus forest, San Juan del Rio—Jalpan de Serra, Pinal Fig. 9. Riparian vegetation, Rio Jalpan, Jalpan de Serra, Queré-

de Amoles, Querétaro. Photo by Erick Daniel Velasco Esquivel. taro. Photo by Erick Daniel Velasco Esquivel.

Amphib. Reptile Conserv. 154 April 2022 | Volume 16 | Number 1 | e308

Cruz-Elizalde et al.

No. 5. Rheohyla miotympanum (Cope, 1863). The Small-

eared Treefrog is a country endemic distributed from “Nuevo

Leon and Coahuila (Sierra Madre Oriental) to Guanajuato

(Sierra Santa Rosa), Hidalgo, and Oaxaca, adjacent Veracruz,

and central Chiapas” (Frost 2020). This individual was

found in Pinal de Amoles, Querétaro. 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 judged

as Near Threatened by the IUCN, but this species has not been

evaluated by SEMARNAT. Photo by Raciel Cruz-Elizalde.

doc ophinst ot apt bead

ae rg see eat

No. 7. Ambystoma velasci (Dugeés, 1888). The Plateau Tiger

Salamander ranges from northwestern Chihuahua southward

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 found in

Pinal de Amoles, Querétaro. Wilson et al. (2013b) determined

its EVS as 10, placing it at the lower limit of the medium

vulnerability category. Its conservation status is considered

as Least Concern by the IUCN, and it has been placed in the

Special Protection (Pr) category by SEMARNAT. Photo by

Raciel Cruz-Elizalde.

Amphib. Reptile Conserv.

ae"

a”

= 2. dij ee ey

Son tinse

No. 6. Smilisca baudinii (Dumeéril and Bibron, 1841). Baudin’s

Treefrog occurs in extreme southern Texas (United States),

and southern Sonora and southwestern Chihuahua (Mexico)

southward (including the Balsas Depression of Mexico) in

the tropical lowlands to Costa Rica on the Pacific slope; Tres

Marias Islands off the coast of Nayarit, Mexico; and seemingly

introduced into Bexar and Refugio counties in southeast-

central Texas, United States (Frost 2021). This individual was

found in Jacalilla, Querétaro. Wilson et al. (2013a) determined

its EVS as 3, placing it in the low vulnerability category. Its

conservation status has been assessed as Least Concern by the

IUCN, but this species is not listed by SEMARNAT. Photo by

Raciel Cruz-Elizalde.

No. 8. Chiropterotriton chondrostega (Taylor, 1941).

The Gristle-headed Splayfoot Salamander is known from

Northwestern Hidalgo and adjacent Querétaro, Mexico, in

cloud forest (at elevations of 1,524—2,042 m asl); and has

also been reported in the state of México (Frost 2021). This

individual was found in Pinal de Amoles, Querétaro. Wilson

et al. (2013b) determined its EVS as 17, placing it in middle

portion of the high vulnerability category. Its conservation

status is considered as Endangered by the IUCN, and it is in

the Special Protection (Pr) category by SEMARNAT. Photo by

Raciel Cruz-Elizalde.

April 2022 | Volume 16 | Number 1 | e308

The herpetofauna of Queretaro, Mexico

Table 1. Monthly minimum, mean (in parentheses), maximum, and annual temperature data (in °C) for the physiographic regions

of Querétaro, Mexico. The localities (and elevation) representing each of the regions are: Central Plateau—Toliman (1,510 m);

Transmexican Volcanic Belt—San Juan del Rio (1,920 m); Sierra Madre Oriental—Jalpan (750 m). Data were taken from Anuario

Estadistico y Geogrdafico de a (INEGI 2017).

Physiographic region

10.2 | 145 | 147

Central Plateau a 5 ke e (19.5) | (21.9) | (23.2)

15.4 | 216 | 229 | 24.7 | 25.6

Transmexican Volcanic eae 13.6

Belt Ag 5 ee 5 he zy (19.5) | (20.5)

16.3 17.4 19.8 22.6 22.9

10.3 15.4 17.6 19.1 17.8

Sierra Madre Oriental (17.9) | (19.7) | (23.4) | (26.1) | (27.6)

20.3 20.5 25.7 26.3 31.3

transformed environments as introduced grasslands

and agricultural areas (Bayona Celis 2016).

Climate

Temperature. Information on the monthly minimum,

mean, and maximum temperatures are given in Table

1 for one locality in each of the three physiographic

regions we recognize in Querétaro. The elevations for

these localities range from 750 m in the Sierra Madre

Oriental (SMO) at Jalpan, to 1,920 m in the Transmexican

Volcanic Belt (TVB) at San Juan del Rio.

The mean annual temperature (MAT) for Jalpan

(elevation 750 m asl) in the SMO is 23.3 °C, while the

MAT for Toliman in the Central Plateau (CP) 1s 19.6 °C,

and the MAT for San Juan del Rio in the TVB is 17.3 °C.

The minimum annual temperatures range from 11.7 °C

in the CP to 17.6 °C in the SMO, the maximum annual

temperatures range from 19.6 °C in the TVB to 25.1 °C

in the SMO, and in the three physiographic regions, the

minimum annual temperatures are 7.5—10.2 °C lower

than the maximum annual temperatures (Table 1). The

mean monthly temperatures peak in May or June (most

often in May), and reach their lowest point in January.

Precipitation. The monthly precipitation is lowest during

the dry season from November to April, and highest during

the rainy season from May to October which includes

(23.2)

264 | 262 | 256 | 235 | 193 | 174 | 178 22.2

(20.1)

226 | 226 | 209 | 199 | 189 | 160 | 158 19.6

wor [owe [oy [oe [ot [sae [oe [om [nr [ne [on

144 | 153 | 155 | 146 | 101 8.5 12.0

(22.3) | (21.6) | (21.0) | (18.8) tee (15.2) | (19.6)

143 | 136 | 13.1 | 13.7 | 11.8 ] 11.0 | 102 11.7

(18.9) | (18.8) | (18.2) | (6.7) | 5.4) | 4.0) | 7.3)

20.0 | 209 | 204 | 209 | 185 | 143 | 15.4 17.6

(26.9)

30.2 | 294 | 291 | 269 | 242 | 192 | 181 25.1

(25.4) | (25.6) | (24.6) | (22.9) | (20.4) | 18.9) | (23.3)

76.0—87.5% (x = 83.5) of the annual precipitation (Table

2). The annual rainfall ranges from 291.3 mm in the CP

to 662.8 mm in the SMO, noting that the latter value is

2.3 times greater than the former (Table 2).

Comments on the Species List

As noted above, we revaluated the status of 17 species

after Cruz-Elizalde et al. (2019) placed them on the

state list. These 17 species were allocated to the three

categories of (1) species currently documented as

occurring 1n Querétaro; (2) species that likely occur in

Querétaro but remain undocumented in the state; and (3)

species that, insofar as we are aware, are not known to

occur in Queretaro.

Eight species whose status was previously in doubt

are now documented as occurring in Querétaro. The

presence of these species in the state, and the literature

that documents their presence, is as follows: (1)

Eleutherodactylus nitidus (recorded by Nieto-Montes de

Oca and Pérez-Ramos [1999]; Informe de CONABIO,

locality Mesa de Leon, el Arbolito, near Hidalgo, in

project H250-CONABIO); (2) Gerrhonotus infernalis

(appears in the H250 report, for which collecting

coordinates are provided); (3) Hemidactylus turcicus

(reported by Tepos-Ramirez et al. [2019]); (4) Scincella

lateralis (reported in the H250 report, based on a

collected specimen); (5) Scincella silvicola (reported

Table 2. Monthly and annual precipitation data (in mm) for the physiographic regions of Querétaro, Mexico. The localities (and

elevation) representing each of the regions are: Central Plateau—Toliman (1,510 m); Transmexican Volcanic Belt—San Juan del

Rio (1,920 m); Sierra Madre Oriental—Jalpan (750 m). Data taken from Anuario Estadistico y Geografico de Querétaro (INEGI

2017). The shaded area indicates the months of the rainy season.

—

(9.1) | (3.4) | (7.2) | 2.2) | @5.4) | 67.4)

260 | 120 | 31.0 | 410 | 875 | 890

Central Plateau (57.1) (51.2) (59.6)

148.0 oi 21.0

(30.6) | (8.4) | (4.5)

1105 | 201 19.0

(334.9)

663.5

33.7 0.0 8.0 1.0 645 | 596 6.5 19 7.5 2.7 29.5

(1.4)1 (1) | 5) | 9.4) | 69.5) | 6.6) | (108.9) | (99.1) | (88.0) | (42.7) | (12.7)

89.5 8.8 11.0 49 | 1267 | 127.4 | 1508 75.8

poke

(542.9)

853.5

Transmexican

Volcanic Belt

(6.4)

166.1 52.1 49.8 9.3

13 2.5 0.0 0.0 5.0 21.6 75.8 78.7 59.1 80.3 0.0 8.4

(8.0) | (10.8) | (0.4) | (27.9) | (45.0) | (23.5) | (155.0) | (149.6) | (195.2) | (94.5) | (25.4) | (15.5)

13.0 | 146 | 13.0 | 321 | 474 | 1464 | 4355 | 1725 | 6450 | 1588 | 37.0 | 249

447.6

(872.2)

1,503.4

Sierra Madre

Oriental

Amphib. Reptile Conserv. 156 April 2022 | Volume 16 | Number 1 | e308

Cruz-Elizalde et al.

Table 3. Composition of the native and non-native ene of Querétaro, Mexico.

p Order | Families | Genera | Species

a EC (NL

po Caudata |

| Subtotal EA

Subtotal

in the H250 report, based on a collected specimen); (6)

Epictia wynni (reported in Querétaro by Wallach [2016]);

(7) Rena dulcis (reported in Querétaro, see The Reptile

Database, http://www.reptile-database.org/; accessed 22

August 2019); and (8) Crotalus polystictus (reported in

Querétaro by Cruz-Pérez et al. [2014]). Thus, these eight

Species are included in the following analyses.

Four species which have not been formally

documented from Querétaro thus far, but are expected to

be documented for the state eventually, are: (1) Coluber

constrictor (recorded in Querétaro by the IUCN, but

with no locality provided); (2) Conopsis biserialis (not

confirmed for the state, but with a high probability of

occurrence, according to the IUCN and Goyenechea

and Flores-Villela [2006]); (3) Ficimia streckeri (a high

probability of occurrence in the state, based on a record

found 3 km from the state line by Lara-Tufifio et al.

[2013]); and (4) Masticophis taeniatus (reported in the

H250 project as having a high probability of occurrence,

but without confirmation). Given the ambiguity of their

actual occurrence in the state at this point, we do not

consider these four species in the subsequent analyses.

Five species reported previously by Cruz-Elizalde

et al. (2019) are not currently believed to occur in

Querétaro, so they are not included in the analyses in this

paper. These species are: (1) Gerrhonotus liocephalus

(According to Good [1994], this species does not occur

in central Mexico, but reports might be based on a

misidentified G. infernalis, also, it is not listed by the

IUCN or in the H250 project as reported in Nieto-Montes

de Oca and Pérez-Ramos [1999]); (2) Lepidophyma

flavimaculatum (recorded in the H250 project, but this

requires confirmation; its distribution appears to lie

farther south in the states of Veracruz and Oaxaca); (3)

Leptodeira maculata (not present in Querétaro based on

its known distribution, and not listed by the IUCN or the

H250 report); (4) Thamnophis marcianus (not present in

Querétaro, neither recorded by Rossman et al. (1996) nor

listed in the state by the IUCN; although it appears in the

H250 report, no locality information or references were

provided); and (5) Crotalus ravus (not known to occur in

Querétaro; its distribution lies farther to the south; this

taxon was not reported for the state by Campbell and

Lamar [2004]). We do not consider these five species in

the subsequent analyses in this paper.

Amphib. Reptile Conserv.

Composition of the Herpetofauna

Families

The herpetofaunal species known to occur in Querétaro

are in 29 families, and include seven families of anurans,

two families of salamanders, 19 families of squamates,

and one family of turtles (Table 3). This total represents

47.5% of the 61 herpetofaunal families known to occur

in Mexico (Wilson et al. 2013a,b). No caecilian or

crocodylian families are represented in the state. Of the

nine amphibian families known to occur in the state,

58.8% of the species (Tables 4—5) are classified in the

Bufonidae (six), Hylidae (eight), and Plethodontidae

(six). Among the 20 remaining herpetofaunal families, 67

(70.5%) of the 95 species are categorized in the families

Phrynosomatidae (12), Colubridae (22), Dipsadidae (15),

Natricidae (eight), and Viperidae (10; see Table 5).

Genera

Seventy-seven herpetofaunal genera are known to occur

in Querétaro, which includes 15 genera of anurans, four

genera of salamanders, 57 genera of squamates, and one

genus of turtles (Table 3). These 77 taxa comprise 35.8%

of the 212 genera known to occur in Mexico (J. Johnson,

unpub. data, 24 December 2020). Among the amphibians

(Table 4), the largest numbers of species are in the genera

Eleutherodactylus (four) and Lithobates (four); among

the reptiles (Table 4), the most speciose genera are

Sceloporus (11), Thamnophis (six), and Crotalus (seven).

Species

The herpetofauna of Querétaro is comprised of 129

species, including 27 anurans, seven salamanders, 92

squamates, and three turtles (Table 3). The current

numbers of native species in these four groups in Mexico

are, respectively, 253, 155, 896, and 51 (J. Johnson,

unpub. data, 24 December 2020). The 129 herpetofaunal

Species in Querétaro represent 9.5% of the 1,361 species

in all of Mexico (J. Johnson, unpub. data, 29 May 2021).

Thus far, one state sharing a common border with

Querétaro has been assessed in the Mexican Conservation

Series—the state of Hidalgo (Ramirez-Bautista et al.

April 2022 | Volume 16 | Number 1 | e308

The herpetofauna of Queretaro, Mexico

Table 4. Distribution of the amphibians, squamates, and turtles of Querétaro, Mexico, by physiographic region. Abbreviations are

as follows: CP = Central Plateau, TVB = Transmexican Volcanic Belt, SMO = Sierra Madre Oriental. See text for descriptions of

these regions. * = species endemic to Mexico, ** = non-native species.

Physiographic regions of Querétaro Number of

Taxon CP TVB SMO regions occupied

Anura (27 species)

Bufonidae (6 species)

Craugastoridae (2 species)

Craugastor augusti

Craugastor decoratus*

Eleutherodactylidae (4 species)

Eleutherodactylus nitidus*

Hylidae (8 species)

Scinax staufferi

Smilisca baudinii

Tlalocohyla godmani*

Tlalocohyla picta

Trachycephalus vermiculatus

Microhylidae (1 species)

Ranidae (4 species)

Zzumae*

Scaphiopodidae (2 species)

Scaphiopus couchii

Spea multiplicata

Caudata (7 species)

Ambystomatidae (1 species)

Plethodontidae (6 species)

Chiropterotriton chondrostega*

Chiropterotriton magnipes*

Chiropterotriton multidentatus*

Squamata (92 species)

Anguidae (4 species)

Gerrhonotus infernalis

——————————

|Anura(27speciesy) |

|Bufonidae(6speciesy |

|Anaxyruspunctatus |

|Anaxyrusspeciosus |

Inciliusnebulifer |

|Craugastoridae(2 species) |

[Craugastoraugusti |

|Craugastordecoratus* |

|Eleutherodactylidae(4 species) |

[Eleutherodactylus guttilatus |

[Eleutherodactyluslongipes* |

Eleutherodactylus nitidus® |

Eleutherodactylusverrucipes* |

|Hylidae(Sspeciesy |

[Rheohyla miotympanum® |

[Scinax stanfferi |

[Smiliscabaudinti |

[Malocohylagodmani® |

Malocohylapicta |

[Trachycephalusvermiculatus |

|Microhylidae(I species) |

|Hypopachusvariolosus |

[Ranidae(4speciesy |

|Scaphiopodidae(2 species) |

[Scaphiopuscouchii |

[Speamultiplicata |

|Caudata(7 species) |

|Ambystomatidae(1 species) |

|Plethodontidae (6 species) |

|Aquiloeuryceacephalica® |

|Aquiloeuryceascandens®* |

|Chiropterotriton chondrostega* | FT

|Chiropterotritonmagnipes* |

|Chiropterotriton multidentatus* |

Usthmura bellii*®

[Squamata(92 species) |

|Anguidae(4 species) |

|Abroniataeniata*® |

[Barisiaciliaris®

|Gerrhonotus infernalis |

|Gerrhonotus ophiurus® |

Gerrhonotus ophiurus*

Amphib. Reptile Conserv. 158 April 2022 | Volume 16 | Number 1 | e308

Cruz-Elizalde et al.

Table 4 (continued). Distribution of the amphibians, squamates, and turtles of Querétaro, Mexico, by physiographic region.

Abbreviations are as follows: CP = Central Plateau, TVB = Transmexican Volcanic Belt, SMO = Sierra Madre Oriental. See text for

descriptions of these regions. * = species endemic to Mexico, ** = non-native species.

Physiographic regions of Querétaro Number of

oe SS so ee

|Corytophanidae(2 species) |

|Corytophaneshernandesii | |

LLaemanctus serratus

|Dactyloidae(I species) |

|Noropssericeus

|Dibamidae(I species) |

Anelytropsis papillosus*

|Gekkonidae(Ispecies) |

|Hemidactylusfrenaus**

Phrynosomatidae (12 species) ff

a a a eS a

Lae —— oi ef

[Sceloporus grammicus 8

[Sceloporusminor®

[Sceloporusparvus*® TE

[Sceloporusscalaris*

[Sceloporusserrifer

|Sceloporus spinosus*

[Sceloporus torquatus*® TT

[Sceloporusvariabilis |

[Scincidae(2speciesy |

|Plestiodoniynxe* |

| Plestiodon tetragrammus TL

|Sphenomorphidae(3species) | |

[Scincellagemmingeri*

[Seincellalateralis

[Scincellasilvicola®

[Teiidae(2species)

|Holcosus amphigrammus® TT

[Xantusiidae(3 species) |

Lepidophymagaigeae* ET

Lepidophymaoceulor®

[Lepidophyma sylvaticum® TL

|Xenosauridae(I species) |

|Xenosaurusmendozai* TT

|Boidae(Ispecies)

|Boaimperator

|Colubridae(22 species) |

Comopsislinewa® ff

|Drymobius margaritiferus |

|Ficimiaolivacea*

|Gyalopioncanum

|Lampropeltispolyzona*

LLampropeltisruthveni®

LLeptophismexicanus TE

|Masticophis mentovarius EH

ut 1

Amphib. Reptile Conserv. 159 April 2022 | Volume 16 | Number 1 | e308

The herpetofauna of Queretaro, Mexico

Table 4 (continued). Distribution of the amphibians, squamates, and turtles of Querétaro, Mexico, by physiographic region.

Abbreviations are as follows: CP = Central Plateau, TVB = Transmexican Volcanic Belt, SMO = Sierra Madre Oriental. See text for

descriptions of these regions. * = species endemic to Mexico, ** = non-native species.

Number of

regions _

$= [——=—=|=—= 5=—

| a

Orybelis porostensis ff

Eee (S(T [A a a

Salvadorabairdi*

[Salvadoragrahamiae TE

———————— = SSS SS SS

[Rosita bocou® [rt

Tantillarubra,

|Trimorphodontau*®

|Dipsadidae(15speciesy) |

|Amastridiumsapperi_

|Chersodromus rubriventris*

|Coniophanesfissidens

|Coniophanespiceivitis TL

|Conophislineaus

Diadophis punctatus | PP

|Geophis mutitorgues*

|Hypsiglenajani 8

Imantodes gemmistratus TL

Leptodeira septentrionalis, |

\Niniadiademata

|Rhadinaeagaigeae* TT

[Tropidodipsas sartorti_

|Elapidae(Ispeciesy)

|Micrurustener

|Leptotyphlopidae(2species) |

JEpictiawynni®

[Renadulcis

|Natricidae(8 species) |

PStoreria hidalgoensis® Ff

an |e |e [es

Thamnophiseques |||. f + fF

anno pins? [Jp

Da Ei a =|

[Typhlopidae(I species) |

[Virgotyphlops braminus®® |

|Viperidae(10 species) |

|Agkistrodontaylori® TE

|Bothropsasper

[Crotalus aquilus® 8

[Crotalusatrox,

|Crotalusmolossus

[Crotalus polystictus®

+]+]4+]+

MM 1

—

Amphib. Reptile Conserv. 160 April 2022 | Volume 16 | Number 1 | e308

Cruz-Elizalde et al.

Table 4 (continued). Distribution of the amphibians, squamates, and turtles of Querétaro, Mexico, by physiographic region.

Abbreviations are as follows: CP = Central Plateau, TVB = Transmexican Volcanic Belt, SMO = Sierra Madre Oriental. See text for

descriptions of these regions. * = species endemic to Mexico, ** = non-native species.

— regions of Querétaro

regions occupied

SMO

[Crotalus totonacus®

[Crotalus triseriatus*

Number of

|Metlapilcoatlus borealis* |

|Kinosternidae(3 species) |

[Kinosternonhirtipes

[Kinosternon scorpioides TE

Total ws to

2020). The herpetofauna of Hidalgo consists of 203

species, which is about 1.6 times the number of species in

Querétaro (129). This proportion is similar to the relative

areas of the two states. The surface area of Hidalgo is

20,813 km? (Ramirez-Bautista et al. 2020) and that of

Querétaro, as noted above, is 11,699 km; therefore,

Hidalgo is 1.8 times the size of Querétaro. Thus, the

state area/species richness ratio is 90.7 for Querétaro

compared to 102.5 for Hidalgo.

Patterns of Physiographic Distribution

A system of three physiographic regions (Fig. 1) was used

to analyze the distribution patterns of the amphibians and

reptiles of Querétaro, and the physiographic distribution

data for the 129 species are tabulated in Table 4 and

summarized in Table 5

The total number of species in each region ranges

from 43 in the Transmexican Volcanic Belt (TVB) to

102 species in the Sierra Madre Oriental (SMO). The

value for the remaining area (Central Plateau) is 64. The

low value of 43 for TVB in Querétaro is 42.2% of the

high value of 102 for SMO in Querétaro. The reason

for this disparity is that the TVB is the smallest of the

three regions in the state, and although it is a significant

montane region in Mexico, it contains less herpetofaunal

diversity than the SMO (Canseco-Marquez et al. 2004;

Flores-Villela et al. 2010), 1.e., 139 vs. 207, respectively.

Four herpetofaunal groups (anurans, salamanders,

squamates, and turtles) are known to occur in Queretaro,

while caecilians and crocodylians have not been recorded

in the state and are not likely to be found in the future.

In three of these groups (anurans, salamanders, and

squamates), the largest number of species occurs in the

SMO; while all three species of turtles are found in the

TVB (Table 5). Twenty-five of the 27 anuran species

(92.6%), all seven of the salamander species (100%), and

69 of the 92 squamates (75.0%) occur in the SMO.

The members of the Querétaro herpetofauna are

distributed in either one, two, or three physiographic

regions as follows: one region (77 of 129 species, 59.7%);

Amphib. Reptile Conserv.

two regions (24, 18.6%); and three regions (28, 21.7%).

The mean regional occupancy is 1.6, which lies outside

the range of 1.9 to 3.7 for the other states examined thus

far in the MCS (Alvarado-Diaz et al. 2013; Mata-Silva et

al. 2015; Johnson et al. 2015a; Teran-Juarez et al. 2016;

Woolrich-Pifia et al. 2016; Nevarez-de los Reyes et al.

2016; Cruz-Saenz et al. 2017; Gonzalez-Sanchez et al.

2017; Lazcano et al. 2019; Ramirez-Bautista et al. 2020).

Of the 129 species found in Querétaro, a large proportion

(101, or 78.3%) occurs in one or two of the three

physiographic regions, which is significant from a

conservation perspective (see below). The number of

species inhabiting a single physiographic region ranges

from eight (in the TVB) to 54 (in the SMO).

The 54 single-region species in the SMO are as follows,

with the numbers referring to the distributional categories

developed by Wilson et al. (2017), country endemics

indicated by single asterisks, and non-native species by

double asterisks:

Anaxyrus speciosus 3

Incilius nebulifer 3

Craugastor decoratus*

Eleutherodactylus longipes*

Eleutherodactylus nitidus*

Scinax staufferi 4

Smilisca baudinii 7

Tlalocohyla godmani*

Tlalocohyla picta 4

Trachycephalus vermiculatus 6

Hypopachus variolosus 7

Scaphiopus couchii 3

Aquiloeurycea scandens *

Chiropterotriton magnipes*

Chiropterotriton multidentatus*

Isthmura bellii*

Abronia taeniata*

Corytophanes hernandezii 4

Laemanctus serratus 4

Norops sericeus 4

April 2022 | Volume 16 | Number 1 | e308

The herpetofauna of Queretaro, Mexico

No. 9. Norops sericeus Hallowell, 1856. The Silky Anole

occurs in the states of Tamaulipas, Hidalgo, San Luis Potosi,

Veracruz, Tabasco, Campeche, Quintana Roo, northern Oaxaca,

Quéretaro, and Puebla. This individual was found in the

municipality of Pisaflores, Hidalgo, near the state of Querétaro.

Wilson et al. (2013a) determined its EVS as 8, placing it in

the low vulnerability category. Its conservation status has been

assessed as Least Concern by the IUCN, but this species is not

listed by SEMARNAT. Photo by Daniel Lara Tufifio.

eet : : ies aa

we 8 Se aye

No. 10. 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 the city of Querétaro,

Querétaro. 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 Raciel Cruz-Elizalde.

No. 11. Sceloporus grammicus Wiegmann, 1828. The Mesquite

Lizard occurs in the states of Chihuahua, Durango, Zacatecas,

Coahuila, San Luis Potosi, Nuevo Leon, Tamaulipas, Oaxaca,

Guerrero, Quéretaro, Hidalgo, Aguascalientes, and Nayarit.

This individual was found in Huimilpan, Querétaro. Wilson

et al. (2013a) determined its EVS as 9, placing it in the low

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

Raciel Cruz-Elizalde.

Amphib. Reptile Conserv.

No. 12. Plestiodon lynxe (Wiegmann, 1834). The Oak Forest

Skink occurs in central and western Mexico, in the states of

Hidalgo, Veracruz, San Luis Potosi, Tamaulipas, Puebla,

Aguascalientes, Querétaro, Guanajuato, Jalisco, and Nayarit.

This individual was found in Pinal de Amoles, Querétaro.

Wilson et al. (2013a) determined its EVS as 10, placing it in the

medium 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 Raciel Cruz-Elizalde.

April 2022 | Volume 16 | Number 1 | e308

Cruz-Elizalde et al.

Table 5. Summary of the distributional occurrence of herpetofaunal families in Querétaro, Mexico, by physiographic province. See

Table 4 for explanation of abbreviations.

amber ot Distributional occurrence

species

CP TVB S

Bufonidae

Craugastoridae

Eleutherodactylidae

Hylidae

Microhylidae

Ranidae

Scaphiopodidae

Subtotal

Ambystomatidae

Plethodontidae

Subtotal

Total

Anguidae

Corytophanidae

Dactyloidae

Dibamidae

Gekkonidae

Phrynosomatidae

Scincidae

Sphenomorphidae

Tetidae

Xantusiidae

Xenosauridae

Subtotal

Boidae

Colubridae

Dipsadidae

Elapidae

Leptotyphlopidae

Natricidae

Typhlopidae

Viperidae

Subtotal

Kinosternidae

Subtotal

Total

Sum total

Anelytropsis papillosus* Lampropeltis polyzona*

Hemidactylus frenatus** Leptophis mexicanus 4

Sceloporus serrifer 4 Mastigodryas melanolomus 4

Plestiodon tetragrammus 3 Oxybelis potosiensis 8

Scincella gemmingeri* Spilotes pullatus 6

Scincella lateralis 3 Adelpicos quadrivirgatum 4

Scincella silvicola* Amastridium sapperi 4

Lepidophyma occulor* Chersodromus rubriventris*

Lepidophyma sylvaticum* Coniophanes fissidens 6

Xenosaurus mendozai* Coniophanes piceivittis 4

Boa imperator 6 Geophis latifrontalis*

Drymobius margaritiferus 8 Imantodes gemmistratus 6

Ficimia olivacea* Ninia diademata 4

Amphib. Reptile Conserv. 163 April 2022 | Volume 16 | Number 1 | e308

The herpetofauna of Queretaro, Mexico

No. 13. Scincella gemmingeri (Cope, 1864). Cope’s Forest

Ground Skink occurs in southeastern Mexico, in Chiapas,

eastern Hidalgo, central and southern Veracruz, Querétaro,

Oaxaca, southward to Tehuantepec, and on the slopes of the

plateau and in lowland areas of Tabasco and Puebla. This

individual was found in Pinal de Amoles, Querétaro. Wilson

et al. (2013a) determined its EVS as 11, placing it in the

medium vulnerability category. Its conservation status has been

regarded as Least Concern by the IUCN, and it has been placed

in the Special Protection (Pr) category by SEMARNAT. Photo

by Raciel Cruz-Elizalde.

No. 14. Xenosaurus mendozai Nieto-Montes de Oca, Garcia-

Vazquez, Zufiga-Vega, and Schmidt-Ballardo, 2013. The

Granular-Scaled Lizard occurs in the states of Querétaro, from

where it was first described, and Hidalgo. This individual was

found in the municipality of Jacala de Ledezma, Hidalgo,

near the state of Querétaro. In this study, and according to

Wilson et al. (2013a), we determined its EVS as 16, placing

it in the high vulnerability category. According to IUCN, its

conservation status is unknown, and this species is not listed by

SEMARNAT. Photo by Christian Berriozabal-Islas.

No. 15. 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, Querétaro, Hidalgo, San Luis Potosi,

Tlaxcala, Veracruz, and Ciudad de México (Ramirez-Bautista et

al. 2014). This individual was found near the city of Querétaro.

Wilson et al. (2013a) determined its EVS as 13, placing it

at the upper limit of the medium vulnerability category. Its

conservation status has been assessed as Least Concern by the

IUCN, but this species is not listed by SEMARNAT. Photo by

Raciel Cruz-Elizalde.

Amphib. Reptile Conserv.

No. 16. Amastridium sapperi (Werner, 1903). The Rusty-

headed Snake occurs in the states of Chiapas, Oaxaca, Hidalgo,

and Querétaro. This individual was found in La Cueva,

Pisaflores, Hidalgo, near the state of Querétaro. Wilson et al.

(2013a) determined its EVS as 10, placing it in the medium

vulnerability category. Its conservation status has been assessed

as Least Concern by the IUCN, but this species is not listed by

SEMARNAT. Photo by Daniel Lara Tufifio.

April 2022 | Volume 16 | Number 1 | e308

Cruz-Elizalde et al.

Tropidodipsas sartorti 4

Storeria hidalgoensis*

Storeria storerioides*

Thamnophis sumichrasti*

Agkistrodon taylori*

Bothrops asper 6

Crotalus totonacus*

Metlapilcoatlus borealis*

Twenty-five of these 54 species (46.3%) are country

endemics and 28 of the remaining 29 are non-endemics,

apart from the single non-native Hemidactylus frenatus.

The distribution ranges of the 28 non-endemic species

are thus: five range to the north, including the United

States; 13 range farther south into Central America; six

range through Central America and into South America;

two range from the United States to Central America; and

two occur from the United States to South America.

The 15 single-region species in the CP are as follows, using

the same asterisk and numbering identifiers as above:

Anaxyrus punctatus 3

Eleutherodactylus guttilatus 3

Sceloporus exsul*

Gyalopion canum 3

Masticophis mentovarius 6

Pseudoelaphe flavirufa 4

Epictia wynni*

Thamnophis pulchrilatus*

Thamnophis scalaris*

Virgotyphlops braminus***

Crotalus atrox 3

Crotalus molossus 3

Crotalus polystictus*

Crotalus scutulatus 3

Crotalus triseriatus*

Six of these 15 species (40.0%) are country endemics

and the remaining nine are non-endemics, except for

the non-native Virgotyphlops braminus. Six of the eight

non-native species also range to the north, including

the United States, while one ranges farther south into

Central America, and another one ranges through Central

America and into South America.

The eight single-region species in the TVB are as follows,

using the same asterisk and numbering identifiers as

above:

Gerrhonotus infernalis 3

Lampropeltis ruthveni*

Salvadora grahamiae 3

Tantilla bocourti*

Leptodeira septentrionalis 8

Thamnophis melanogaster*

Kinosternon hirtipes 3

Kinosternon scorpioides 6

Three of these eight species (37.5%) are country endemics

and the remaining five are non-endemics. Three of these

five species also range to the north, including the United

States, one species ranges through Central America and

into South America, and one species occurs from the

United States to South America.

In summary, of the 77 single-region species found in

Querétaro, 34 (44.2%) are country endemics, 41 (53.2%)

are non-endemics, and two are non-natives (2.6%). Of

the three physiographic regions in the state, the SMO

is of greatest conservation significance, inasmuch as it

encompasses the largest overall number of species (102),

the largest number of single-region species (54), and the

largest number of country endemics (25).

A Coefficient of Biogeographic Resemblance (CBR)

matrix was assembled for examining the herpetofaunal

similarity relationships of the three physiographic regions

in Querétaro (Table 6). The SMO contains the most

species richness (102 species) and the TVB the least (43

species). The mean species richness value for all three

regions is 69.7. The number of shared species between

each of the regional pairs ranges from 31 between the

TVB and SMO regions to 45 between the Central Plateau

and the SMO. The mean value of shared species among

all three regions is 36.0.

The CBR values in Table 6 range from 0.43 to 0.59.

The lowest value is that between the Sierra Madre Oriental

and the Transmexican Volcanic Belt. These two regions

lie at opposite extremes in the state (Fig. 1). The highest

value is that between the Transmexican Volcanic Belt

and the Central Plateau, which are contiguous regions

Table 6. Pair-wise comparison matrix of Coefficient of Biogeographic Resemblance (CBR) data of the herpetofaunal relationships

for the three physiographic regions in Querétaro, 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. 10 for the UPGMA dendrogram produced

from the CBR data.

Central Plateau

Central Plateau 64

Transmexican Volcanic Belt 0.60

Sierra Madre Oriental 0.54

Amphib. Reptile Conserv.

Transmexican Volcanic Belt

Sierra Madre Oriental

32 45

43 31

0.43 102

April 2022 | Volume 16 | Number 1 | e308

The herpetofauna of Queretaro, Mexico

Bie 2 4

)

celllpteee || eats

= =

aa) i)

— a

2 n

[—) i)

> o—_

| =

Z| ¢ fla a ¥

x rf

2 =

e| 2

Fig. 10. UPGMA-generated dendrogram illustrating the simi- x

larity relationships of species richness among the herpetofaunal E y

components in the three physiographic regions of Querétaro = F

(based on the data in Table 7; Sokal and Michener 1958). Simi- zs oe

larity values were calculated using Duellman’s (1990) Coef- 5

ficient of Biogeographic Resemblance (CBR).

located in the south-central part of the state. The overall eg

CBR values among the three physiographic regions are 5 E a A 8

as follows, arranged from the highest to the lowest value O §

(with species numbers in parentheses):

Transmexican Volcanic Belt (43) — 0.59 — Central Plateau (65)

Sierra Madre Oriental (102) — 0.54 — Central Plateau (65)

ala un uw

Based on the data in Table 6, a UPGMA dendrogram HS pales Ao

(Fig. 10) was developed to illustrate the herpetofaunal pe

resemblance patterns among the three physiographic

regions of Querétaro (Fig. 1). The diagram demonstrates r

that two regions in Querétaro, the CP and the TVB, share 5 s ee ee

the higher herpetofaunal resemblance (0.60 level). Both Z. 3

of these regions are largely montane and broadly contact

one another in the southern portion of the state. These

two regions are more distinguished (0.43 level) from the S 2

Sierra Madre Oriental (SMO). 5 E E Sk

As indicated in the Introduction, we consider it useful S 5

to compare the herpetofaunal representation of the two x

physiographic regions in Querétaro with those of the =

same two regions represented in Puebla (Woolrich-Pifia es x)

et al. 2017) and Hidalgo (Ramirez-Bautista et al. 2020). BE] | m a

We placed the comparative data in Table 7. Most of the A E

species in the two physiographic regions we examined

are either Mexican endemic or non-endemic species,

; ; ; ' > Z

while few state endemics and non-native species occur f=

: . : : = & te &

in either region. As expected, the number of species ay pS

in the two regions examined increases along with the U5

size of the herpetofauna in each of the three states, L.e.,

from Querétaro (129 species) to Hidalgo (202 species)

to Puebla (267 species). In each of the three states the

number of country endemics is greater than the number

of non-endemics. The proportions of country endemics

Table 7. Species comparison of three distributional categories recorded in the two physiographic regions common to the three states of Querétaro, Hidalgo, and Puebla in Mexico.

Querétaro

Hidalgo

Puebla

Amphib. Reptile Conserv. 166 April 2022 | Volume 16 | Number 1 | e308

Cruz-Elizalde et al.

compared to the regional totals for the Sierra Madre

Oriental in each of the three states are similar (55.9%,

52.7%, and 55.1% for Querétaro, Hidalgo, and Puebla,

respectively). Likewise, each of the proportions for the

Transmexican Volcanic Belt are reasonably similar to

one another (53.5%, 69.4%, and 61.6%, respectively).

The proportions of the non-endemics compared to

the regional totals for the Sierra Madre Oriental in the

same three states also are similar to one another (43.4%,

44 8%, and 42.7%, respectively), whereas those for the

Transmexican Volcanic Belt are less consistent (46.5%,

23.5%, and 36.8%, respectively).

Distribution Status Categorizations

In discussing the distribution status of the members of the

Querétaro herpetofauna, we used the system developed by

Alvarado-Diaz et al. (2013) which was used in all the other

entries of the Mexican Conservation Series (see above).

The categories in this system are non-endemic, country

endemic, state endemic (only Sce/oporus exsul), and non-

native. The categorizations for each species are listed in

Table 8 and these data are summarized in Table 9.

The numbers of species in each of the three applicable

categories, in decreasing order, are: country endemics,

67 (51.5%); non-endemics, 60 (46.2%); and non-natives,

3 (2.3%). As with the states of Michoacan (Alvarado-

Diaz et al. 2013), Nayarit (Woolrich-Pifia et al. 2016),

Jalisco (Cruz-Saenz et al. 2017), Puebla (Woolrich-Pifia

et al. 2017), and Hidalgo (Ramirez-Bautista et al. 2020),

the largest number of herpetofaunal species in Querétaro

is in the country endemic category. In other states, the

largest number falls within the non-endemic category,

1.e., Oaxaca (Mata-Silva et al. 2015), Tamaulipas (Teran-

Juarez et al. 2016), Nuevo Leon (Nevarez-de los Reyes et

al. 2016), and Chiapas (Johnson et al. 2015a).

Only one endemic species occurs in Querétaro

(Sceloporus exsul), and in the 10 previous individual-

state entries in the Mexican Conservation Series the

number of state endemics was found to be variable,

ranging from one in Nayarit and Nuevo Leon (Woolrich-

Pifia et al. 2016; Nevarez-de los Reyes 2016) to 93 in

Oaxaca (Mata-Silva et al. 2015).

Two non-native species are found in Querétaro,

Hemidactylus frenatus and Virgotyphlops braminus.

These two taxa are the most widespread of the non-

native species recorded in the 12 entries in the Mexican

Conservation Series (Ramirez-Bautista et al. 2020), and

as of this contribution, they now have been recorded in

11 and 12 states or tri-state regions, respectively.

Wilson et al. (2017) developed a system for

categorizing the distribution of the herpetofauna of

Mesoamerica, and applying those categories to this

study, the data are summarized in Table 10. Previously,

we noted that 67 species in Querétaro are endemic to

Mexico, and thus 60 native species are not. These 60

species are allocated to five of the categories established

Amphib. Reptile Conserv.

by Wilson et al. (2017): MXUS, MXCA, MXSA, USCA,

and USSA. As expected, the largest number of species

falls into the MXUS category (26, 43.3%), which is

followed by MXCA (17, 28.3%), MXSA (9, 15.0%),

USCA (5, 8.3%), and finally USSA (3, 5.0%).

Principal Environmental Threats

In this section we discuss the problems affecting the

sustainability of the amphibian and reptile populations in

Querétaro that we consider to be of greatest significance.

Several negative factors apply, such as the increasing and

unregulated clearing of forests for farming and raising

livestock (for grazing areas), the construction of roads,

the constant and increasing pollution in bodies of water,

emerging diseases, forest fires, and strongly ingrained

cultural factors (Cruz-Elizalde et al. 2016, 2019).

Humans have caused all these factors, either directly or

indirectly, so they should be considered “anthropogenic

effects.”

Deforestation. Despite the fact that Querétaro is

substantially covered with vegetation, primarily in the

northern part of the state where oak forests, pine forests,

tropical vegetation, and cloud forest still remain, many of

these areas have been highly deforested for their timber

resources. Sawmills for wood extraction are common

in many places, and woodlands often are transformed

into areas for agriculture and livestock use. Forested

areas in the state consist of 737,821 ha, of which 51.4%

corresponds to arid areas such as chaparral and shrubland,

24.1% to temperate forest dominated by conifers, 9.1%

to various forests (low, medium, and high), and 15.1% to

disturbed areas with various degrees of effect, without

vegetation cover, or of little importance.

The areas of natural vegetation, whether with forests

or other types of cover, have been deforested to create

agricultural areas, urban settlements, industrial parks,

gas pipelines, aquaculture, or roads (Fig. 11). About 80%

of all forest fires have been intentional. Accordingly,

federal agencies such as the National Forestry

Commission (CONAFOR) and state agencies such as

the Forest Department of the Querétaro Secretariat for

Agricultural Development (SEDEA) have devoted

resources, personnel, and campaigns to the mitigation

of these fires. In addition, reforestation programs have

focused on regions with temperate forest sites, where

large portions of the area have been reforested, such as

in the municipalities of Cadereyta de Montes, Colon, and

Pefiamiller, where 1,555 ha in these three municipalities

have been reforested since 2016.

Deforestation has been more extensive in areas of

northern Querétaro, both in temperate environments

and forests. This destruction has largely affected the

amphibians and reptiles that primarily inhabit these

environments, such as salamanders of the genera

Chiropterotriton and Pseudoeurycea; anurans such

April 2022 | Volume 16 | Number 1 | e308

The herpetofauna of Queretaro, Mexico

No. 17. Thamnophis eques (Reuss, 1834). The Mexican Garter

Snake occurs in Mexico from the vicinity of the Pico de

Orizaba northwestward to and south to Aguascalientes, Sonora

and Chihuahua, Quéretaro, Oaxaca, Nuevo Leon, Hidalgo,

San Luis Potosi, Guanajuato, Puebla, Guerrero, Nayarit, and

Morelos. This individual was found in the locality of Mesa de

Leon, Querétaro. Wilson et al. (2013a) determined its EVS as

8, placing it in the low vulnerability category. Its conservation

status has been assessed as Least Concern by the IUCN,

and it has been placed in the Endangered (A) category by

SEMARNAT. Photo by Raciel Cruz-Elizalde.

ee ee

Coralsnake occurs “from the Mississippi River westward into

Texas, in the United States, and in Mexico, from Tamaulipas

south to Veracruz...” (Lemos-Espinal and Dixon 2013: 240).

This individual came from Zona Metropolitana de Querétaro in

the municipality of El Marques. Wilson et al. (2013a) calculated

its EVS as 11, placing it in the medium vulnerability category.

Its conservation status has been determined as Least Concern

by IUCN, and this species is not listed by SEMARNAT. Photo

by Diego Baez.

Amphib. Reptile Conserv.

No. 18. Thamnophis sumichrasti (Cope, 1866). Sumichrast’s

Garter Snake occurs in the states of Oaxaca, Chiapas, Querétaro,

San Luis Potosi, Tabasco, Puebla, Veracruz, and Hidalgo. This

individual was found in Landa de Matamoros, Querétaro.

Wilson et al. (2013a) determined its EVS as 15, placing it in

the high vulnerability category. Its conservation status has been

assessed as Least Concern by the IUCN, and it has been placed

in the Endangered (A) category by SEMARNAT. Photo by

Raciel Cruz-Elizalde.

No. 20. Crotalus aquilus Klauber, 1952. The Dusky Rattlesnake

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 in the Area Natural

de Pefia Colorada, in the muncipality of Querétaro. Wilson et

al. (2013a) ascertained its EVS as 16, placing it in the middle

portion of the high vulnerability category. This species has been

assessed as Least Concern by IUCN, and placed in the Special

Protection (Pr) category by SEMARNAT. Photo by Alejandro

Peralta Robles.

April 2022 | Volume 16 | Number 1 | e308

Cruz-Elizalde et al.

Table 8. Distributional and conservation status measures for members of the herpetofauna of Querétaro, Mexico. Distributional

Status: CE = endemic to country of Mexico; NE = not endemic to state or country; and NN = non-native. The numbers suffixed

to the NE category signify the distributional categories developed by Wilson et al. (2017) and implemented in the taxonomic

list at the Mesoamerican Herpetology website (http://mesoamericanherpetology.com), as follows: 3 (species distributed only in

Mexico and the United States); 6 (species ranging from Mexico to South America); 7 (species ranging from the United States to

Central America); and 8 (species ranging from the United States to South America). Environmental Vulnerability Score categories

(taken from Wilson et al. 2013a,b): low (L) vulnerability (EVS of 3-9); medium (M) vulnerability (EVS of 10-13); and high (H)

vulnerability (EVS of 14-20). IUCN categories: 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 Alvarado-Diaz et al. (2013), Johnson et al. (2015a), and Mata-Silva et al. (2015)

for explanations of the EVS, IUCN, and SEMARNAT rating systems.

Distributional | E2vironmental IUCN SEMARNAT

Taxon Vulnerability aiey-

Status categorization status

Category (score)

H (14)

L (5)

M (12)

L (6)

M (11)

L (3)

L (8)

H (15)

M (11)

H (15)

M (12)

H (16)

L(7)

M (10)

L(9)

L(4)

L(3)

M (13)

L (8)

L(4)

Fcc co

QATATIQIaA

ZiZ\Z\izZ

RnINnINIA

ee | Se Se ey te

QTOIGIQI|CIaQ|aq}|oa

ZIZ)oIZ|2/A2lo/Al|a

RIAA IALNIAIFA LAIN

Scinax staufferi

Smilisca baudinii

Tlalocohyla godmani*

Tlalocohyla picta

Cil<

Trachycephalus vermiculatus

Hypopachus variolosus

Lithobates montezumae*

M (13)

M (12)

L(3)

L (6)

M (10)

H (14)

H(17)

H (17)

H (16)

H (15)

M (12)

H (15)

H (14)

M (13)

M (12)

M (13)

L (8)

M (10)

tay

ine}

Lithobates neovolcanicus*

Lithobates spectabilis*

(S)

ro)

Scaphiopus couchii

ine}

Ambystoma velasci*

ee)

Aquiloeurycea cephalica*

Aquiloeurycea scandens*

Chiropterotriton chondrostega*

ine}

Chiropterotriton multidentatus*

Isthmura bellii*

Abronia taeniata*

Barisia ciliaris*

Chiropterotriton magnipes*

Spea multiplicata

Zo |

Nyala

Amphib. Reptile Conserv. 169 April 2022 | Volume 16 | Number 1 | e308

The herpetofauna of Queretaro, Mexico

Table 8 (continued). Distributional and conservation status measures for members of the herpetofauna of Querétaro, Mexico.

Distributional Status: CE = endemic to country of Mexico; NE = not endemic to state or country; and NN = non-native. The

numbers suffixed to the NE category signify the distributional categories developed by Wilson et al. (2017) and implemented in the

taxonomic list at the Mesoamerican Herpetology website (http://mesoamericanherpetology.com), as follows: 3 (species distributed

only in Mexico and the United States); 6 (species ranging from Mexico to South America); 7 (species ranging from the United

States to Central America); and 8 (species ranging from the United States to South America). Environmental Vulnerability Score

categories (taken from Wilson et al. 2013a,b): low (L) vulnerability (EVS of 3—9); medium (M) vulnerability (EVS of 10-13); and

high (H) vulnerability (EVS of 14-20). IUCN categories: 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 Alvarado-Diaz et al. (2013), Johnson et al. (2015a), and Mata-Silva

et al. (2015) for explanations of the EVS, IUCN, and SEMARNAT rating systems.

Distributional | E2vironmental IUCN SEMARNAT

Taxon Vulnerability aiey-

Status categorization status

Category (score)

M (12)

M (13)

H (17)

L()

H (14)

H (15)

M (12)

L(©)

M (12)

M(11)

L(G)

M (10)

M (12)

M (11)

M (13)

M (12)

L()

M (11)

M (13)

H (14)

M (11)

(2 Se ISIS SEIS ISIE IE IEBIRIPIGIPleic

GI MJQALALAJSAJAJALALAIJAQIJasaQsaqQjalsayAjayjajoa

SIZIZ|> [el e(Z/~e(ZIZ/Z/Z(Z/ZIZ) ol, | ZZ

TININ TIT IDNIT [NINININININIM!]™ Nin

H (16)

M (10)

M (13)

M (11)

L©)

L@)

L(@)

M (11)

H (16)

L©)

L(©)

M (13)

L©)

L(G)

H (14)

M (10)

H (15)

M (10)

L©)

Yi) ced cd 1 al td al (ca tetas

Z|zZ z\z|z\z|z/z/z|z

Nn NIMNINININININR

emt Reed Lend lew

Ql[aAlal1a

Z|>(Zl>]Z

Nit1n N

NE NS

Amphib. Reptile Conserv. 170 April 2022 | Volume 16 | Number 1 | e308