Official journal website:

amphibian-reptile-conservation.org

Amphibian & Reptile Conservation

15(1) [Taxonomy Section]: 1-35 (e272).

urn:lsid:zoobank.org:pub:9B3E8106-74E8-428F-BOBB-3CCD9EFDFOF3

Two new species of Eleutherodactylus (Anura:

Eleutherodactylidae) from Southern Mexico, with comments

on the taxonomy of related species and their advertisement

calls

123."Christoph I. Griinwald, **Jacobo Reyes-Velasco, **Héctor Franz-Chavez, **Karen I. Morales-

Flores, ?*lvan T. Ahumada-Carrillo, 2*4Christopher M. Rodriguez, and 2°Jason M. Jones

'Biencom Real Estate, Carretera Chapala—Jocotepec #57-1, C.P. 45920, Ajijic, Jalisco, MEXICO *Biodiversa A.C., Avenida de la Ribera #203,

C.P. 45900, Chapala, Jalisco, MEXICO *Herp.mx A.C., Villa de Alvarez, Colima, MEXICO 4Los Angeles Zoo and Botanical Gardens, 45333 Zoo

Drive, Los Angeles, California 90027, USA

Abstract.—An analysis of morphological and molecular data is presented here, as well as notes on

advertisement calls from populations of the Eleutherodactylus nitidus species group (subgenus Syrrhophus)

from southern Mexico, and two new species are described based on the results. Eleutherodactylus

maculabialis sp. nov. is unique among its congeners by a combination of characters including widely

expanded finger tips, indistinct but visible inguinal glands, a dark venter, and conspicuous orange or pale

cream spots present on the upper lip. E/eutherodactylus sentinelus sp. nov. is unique among its congeners

by the combination of its advertisement call, smooth dorsal and ventral skin, and a unique combination

of a brown dorsum, pale interorbital bar and mid-dorsal stripe, and bright yellow inguinal flash coloration.

The molecular phylogenetic analysis indicates that both species are part of the Eleutherodactylus nitidus

species group, and closely related to one another. Known collecting localities for both new species suggest

they are restricted to small ranges in moist pine-oak woodland, cloud forest, and oak-tropical deciduous

forest on the windward slopes of the Sierra Madre del Sur in Guerrero. We discuss the relationships of the

new species to all the species in the Eleutherodactylus nitidus species group and the validity of some related

taxa based on the results of our phylogenetic analysis. The male advertisement calls of the new species are

presented graphically, along with those of the closely related species occurring in the Sierra Madre del Sur.

Range maps are presented for all species of the E/eutherodactylus nitidus species group, as the group is

currently understood.

Keywords. Amphibia, common names, conservation, Guerrero, phylogenetics, Sierra Madre del Sur

Resumen.—Presentamos un analisis de datos morfologicos, moleculares y de cantos de poblaciones del

grupo Eleutherodactylus nitidus (subgénero Syrrophus) del sur de Mexico y describimos dos nuevas especies

basado en nuestros resultados. Eleutherodactylus maculabialis sp. nov. es unica entre sus congeéneres por

una combinacion de caracteres, incluyendo la punta de los dedos ampliamente expandida, la precencia de

glandulas inguinales poco distinguibles y presentar coloraciOn con un fondo oscuro y manchas naranjas

oO palidas en el labio superior. Eleutherodactylus sentinelus sp. nov. es unica entre sus congéneres por

la siguiente combinacion de caracteres que incluyen su canto de advertencia, piel lisa en el dorso y zona

ventral y una combinacion unica de coloracion dorsal café, una barra interorbital palida, una linea a lo

largo del dorso y una coloracion inguinal amarillo brillante. El analisis filogenético molecular indica que

ambas especies pertenecen al grupo Eleutherodactylus nitidus, y estan cercanamente relacionadas. Las

localidades de colecta conocidas para ambas especies sugieren que las mismas tienen una distribucion

restringida a pequenos rangos en bosques humedos de pino-encino, bosque mesofilo de montana y bosque

tropical cauducifolio con encinos en las pendientes de la Sierra Madre del Sur de Guerrero. Discutimos la

relacion entre todas las especies asociadas al grupo Eleutherodactylus nitidus asi como la validez de algunos

taxones relacionados basado en nuestro analisis filogenetico. Analizamos los cantos de advertencia de las

nuevas especies y las especies relacionadas que ocurren en la Sierra Madre del Sur. Finalmente presentamos

mapas de distribucion de todas las especies del grupo Eleutherodactylus nitidus, ya que actualmente no se

entiende bien dicho grupo.

Palabras clave. Anfibios, conservacion, filogenética, Guerrero, Sierra Madre del Sur

Correspondence. ‘cgruenwald@switaki.com

Amphib. Reptile Conserv. 1 February 2021 | Volume 15 | Number 1 | e272

Two new Eleutherodactylus species from Mexico

Citation: Griinwald Cl, Reyes-Velasco J, Franz-Chavez H, Morales-Flores KI, Ahumada-Carrillo IT, Rodriguez CM, Jones JM. 2021. Two new species

of Eleutherodactylus (Anura: Eleutherodactylidae) from Southern Mexico, with comments on the taxonomy of related species and their advertisement

calls. Amphibian & Reptile Conservation 15(1) [Taxonomy Section]: 1-35 (e272).

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 November 2020; Published: 13 February 2021

Introduction

The frogs of the genus Eleutherodactylus Dumeril

and Bibron, 1841 are among the most diverse and

taxonomically challenging groups of amphibians in the

New World (Hedges et al. 2008). The genus consists of

five subgenera (Hedges et al. 2008), four of which are

native to the West Indies (Eleutherodactylus, Euhyas

Fitzinger 1843; Pelorius Hedges 1989; and Schwartzius

Hedges et al. 2008), and a fifth (Syrrhophus Cope 1878),

which is native to continental North America and Cuba.

The native range of Syrrhophus extends from west-central

Texas and central Sonora south through Mexico and into

Guatemala and Belize, with two species 1n western Cuba.

One species, E. campi, has been introduced to some

urbanized parts of Texas (Dixon 2000), Louisiana (Hardy

2004), Alabama (McConnell et al. 2015), and Arizona (D.

Ortiz, in press), presumably via the horticultural trade.

Whereas the systematics of the West Indian subgenera

has been relatively well studied (Hedges et al. 2008),

less attention has been given to the systematics of the

subgenus Syrrhophus until recently. A recent effort

by the authors sampled all of the currently recognized

species of Syrrhophus in the United States, Mexico, and

Guatemala. This resulted in Reyes et al. (2015) describing

two distinctive new species of Eleutherodactylus from

western Mexico based on mitochondrial DNA (mtDNA)

sequence data, morphology, and advertisement calls.

In a continuation of that study, Grtinwald et al.

(2018) analyzed the morphological and molecular data

of all known species within the subgenus, as well as the

taxonomic history of Syrrhophus. That review led to the

assignment of the continental North American species

to two species series: the Eleutherodactylus longipes

species series, which contains only the Eleutherodactylus

longipes species group; and the Eleutherodactylus nitidus

species series, which contains the Eleutherodactylus

nitidus species group and the Eleutherodactylus modestus

species group.

Those authors reviewed the Eleutherodactylus

modestus species group in detail, describing six new

species, synonymizing E. nivicolimae (Dixon and Webb,

1966) with E. rufescens (Duellman and Dixon, 1959),

and providing data for the distinctiveness of E. modestus

(Taylor, 1942), FE. pallidus (Duellman, 1958), E. teretistes

(Duellman, 1958), and E. wixarika Reyes-Velasco et al.,

2015. Furthermore, those authors presented molecular

evidence that E. orarius (Dixon, 1957) is distinct from

E. nitidus (Peters, 1870). Recently, Palacios-Aguilar

and Santos-Bibiano (2020) described a new species

of saxicolous Eleutherodactylus from the lowlands of

Guerrero, near the town of Tierra Colorada. With that

Amphib. Reptile Conserv.

addition, there are currently 34 recognized species in the

subgenus Syrrhophus in continental North America, plus

two more in Cuba.

Here, we review the Eleutherodactylus nitidus species

group in detail, and describe two more new species in

that species group (sensu Griinwald et al. 2018) from

the state of Guerrero. The morphological and molecular

data for all named continental species of the subgenus

Syrrhophus are analyzed to define the two new species.

In addition, two subspecies are elevated to the species

level and two other species are synonymized based on

the results. These rearrangements result in 37 named

species recognized as valid in the subgenus Syrrhophus,

with (potentially) more awaiting description pending the

completion of ongoing investigations.

Materials and Methods

Taxonomic sampling. From 2003-2019, the authors

collected specimens of all known species of the subgenus

Syrrhophus in Mexico, the United States, and Central

America (Reyes-Velasco et al. 2015; Grtinwald et al.

2018). Specimens of all currently recognized species of

the subgenus (Frost 2020) were examined, and specimens

of most species were measured, with the exceptions of

E. verruculatus (Peters 1870) and the recently described

E. erythrochomus (Palacios-Aguilar and Santos-Bibiano

2020). Comparisons of the new species described herein

with the recently described species used the photos and

data provided by the authors in the description (Palacios-

Aguilar and Santos-Bibiano 2020). The validity of E.

verruculatus has been questioned by various authors

(Firschein 1954; Lynch 1970; Grunwald et al. 2018), and

it is currently considered to be an enigmatic name with

no known locality, genetic material, or recently collected

specimens available.

All frogs were photographed alive—including dorsal,

lateral, and ventral profiles, as well as photographs of

each one showing the colors of flanks and flash colors on

the groin and thigh. The frogs were euthanized with 10%

ethanol or with topical benzocaine and tissue samples

were obtained from thigh muscle or liver upon death and

preserved in 96% ethanol. Specimens were preserved in

10% formalin and transferred to 70% ethanol for storage.

Measurements were taken from additional specimens

of the subgenus Syrrhophus in the Museo de Zoologia,

Facultad de Ciencias (MZFC) of Universidad Nacional

Autonoma de México (UNAM) and in the Amphibian

and Reptile Diversity Research Center (ARDRC) of the

February 2021 | Volume 15 | Number 1 | e272

Grunwald et al.

University of Texas at Arlington (UTA). Measurements

were not taken directly for the type specimens of

some previously described taxa so this study used

the measurements of the type specimens provided in

their original descriptions and published literature.

Measurements of Eleutherodactylus dilatus (Davis and

Dixon 1955), E. fuscus (Davis and Dixon 1955) (= E.

maurus), and E. albolabris (Taylor 1943) are given in

their original descriptions and in Dixon (1957a,b).

Measurements for EF. nitidus (Peters 1870) and E. petersi

(Duellman 1954) were taken from Dixon (1957a,b),

while measurements for E. orarius (Duellman 1958)

and EF. syristes (Hoyt 1965) were taken from their

original descriptions. For Eleutherodactylus pipilans

(Taylor 1940), FE. nebulosus (Taylor 1943), and “E.

rubrimaculatus” (Taylor and Smith 1945) this study used

the measurements provided in their original descriptions

and in Lynch (1970). Measurements of each of the above

species were also taken from specimens collected during

this study from the type localities or nearby.

The material collected 1s deposited at the Museo

de Zoologia, Facultad de Ciencias (MZFC) of the

Universidad Nacional Autonoma de México (UNAM)

in Mexico City. Specimen numbers for all material

examined are provided in Appendix 1. While the

specimens collected were formally catalogued at the

MZEC, several of the examined specimens from both the

MZFC and UTA collections have not been catalogued,

and in such cases the original field numbers and their

respective museums are listed.

Morphological measurements. The characters and

terminology used herein follow those of Lynch and

Duellman (1997), Savage (2002), and Griinwald et al.

(2018). The following measurements were taken for

each specimen (abbreviations listed in parentheses):

snout-vent length (SVL); head length (HL); head width

(HW); eyelid width (EW); interorbital distance (IOD),;

internarial distance (IND); eye-naris distance (END);

diameter of eye (ED); width of tympanum (TW); height

of tympanum (TH); eye-tympanum distance (ETD);

upper arm length (UpL); forearm length (FoL); hand

length (HaL); length of 1‘ finger (FIL); width of pad on

1s finger (F1PW); width of 1“ finger (F1W); length of 2"

finger (F2L); width of pad on 2™ finger (F2PW); width of

2" finger (F2W); length of 3" finger (F3L); width of pad

on 3" finger (F3PW); width of 3" finger (F3W); length

of 4" finger (F4L); width of pad on 4" finger (F4PW),;

width of 4" finger (F4W); inner palmar tubercle length

(IPTL); middle palmar tubercle length (MPTL); outer

palmar tubercle length (OPTL); femur length (FeL); tibia

length (TL); tarsal length (TaL), foot length (FL), total

foot length (TotFL); length of 2"! toe (T2L); width of

pad on 2™ toe (T2PW):; width of 2™ toe (T2W); length

of 3 toe (T3L); width of pad on 3 toe (T3PW); width

of 3™ toe (T3W); length of 4" toe (T4L); width of pad

on 4" toe (T4PW); width of 4" toe (T4W); length of 5"

Amphib. Reptile Conserv.

toe (TS5L); width of pad on 5" toe (TSPW); width of 5"

toe (TSW); inner metatarsal tubercle length (MTL); and

outer metatarsal tubercle length (OMTL). Hand length

(HA) was measured from the tip of the longest finger to

the base of the palm, and foot length (FL) was measured

from the tip of the longest toe to the base of the tarsus.

The outer palmar tubercle refers to a small tubercle on

the outer surface of the palm, but not one of the larger

supernumerary tubercles. While these tubercles are

usually present in Syrrhophus, they are generally absent

in some species and their presence is variable in others.

Molecular analysis: DNA extraction and PCR

amplification. A detailed description of the DNA

extraction and PCR amplification protocols can be found

in Griinwald et al. (2018). DNA was extracted from tissue

samples using a standard potassium acetate protocol

and a fraction of the 16s rRNA mitochondrial gene was

sequenced using either the primers LX12SN 1a (forward)

and LX16S1Ra (reverse) of Zhang et al. (2013), or

the modified primers 16Sar and 16Sbr of Bossuyt and

Milinkovitch (2000). Unpurified PCR products were

then shipped for sequencing to BGI Tech Solutions

(Hong Kong).

Molecular analysis: sequence alignment and

phylogenetic analysis. Regions with poor quality base

calls were removed by manually trimming the 5’ and

3’ ends of all sequences using the program Geneious

v6.1.6 (Biomatters Ltd., Auckland, New Zealand). All

sequences were then aligned in Muscle (Edgar 2004),

with a final alignment of 560 base pairs. Additionally,

multiple sequences already available 1n GenBank

were included. Two Cuban species of the subgenus

Syrrhophus (E. symingtoni and E. zeus) were included

as outgroups in the phylogenetic analysis. The final

alignment included 104 individuals, of which 65 are

new. All the new sequences have been deposited in

GenBank and their accession numbers are shown in

Appendix 2.

Bayesian inference of phylogeny (BI) was

performed in MrBayes v3.2.2 (Ronquist et al.

2012), implemented on the CIPRES_ Science

gateway server (Miller et al. 2010). First, the best-

fit models of nucleotide substitution for the 16s

rRNA mitochondrial gene were selected using the

Bayesian Information Criterion (BIC) implemented

in PartitionFinder v1.1.1 (Lanfear et al. 2012). The

Bayesian analysis consisted of four runs of 10 million

generations each, with four chains (one cold and

three heated), and sampling every 1,000 generations.

Tracer v1.6 (Drummond and Rambaut 2012) was

used to confirm the convergence of the independent

runs, based on overlap in likelihood and parameter

estimates among runs, as well as effective sample

size (ESS) and Potential Scale Reduction Factor value

estimates (PSRF). PSRF indicated that individual runs

had converged by 100,000 generations, so the first

25% of the runs were discarded as burn-in. Finally,

posterior probability values were annotated on the

February 2021 | Volume 15 | Number 1 | e272

Two new Eleutherodactylus species from Mexico

resulting topology using the program TreeAnnotator

v1.8.3 (Rambaut et al. 2014). Additionally, genetic

distances were obtained for the members of the group

through the use of Mega X (Kumar et al. 2018).

Advertisement call graphing. Vocalizations were

recorded for two individuals of each new species

described here, as well as all other members of the

Eleutherodactylus (Syrrhophus) nitidus — species

group (sensu Grinwald et al. 2018). The frogs were

recorded while they were actively calling in the field,

using the WavePad free recording software (NCH

Software 2015) on various Apple iPhones. The calls

were recorded at distances ranging from 50-150 cm,

although distances within 100 cm of the frog were

used whenever possible. Ambient temperatures were

not obtained at the time of recording, but the time of

day when each recording was made was noted.

The individual calls were isolated from other calls

and background noise using Adobe Audition CC, and

the calls were then analyzed using the software Raven

Pro 1.5 (The Cornell Lab of Ornithology 2014). The

Raven Pro settings were as follows: window size

= 256 samples; window type = Hanning; overlap =

50%; DFT size = 256 samples; and grid spacing =

188 Hz. Temporal and spectral properties of isolated

calls were analyzed using the Seewave version

1.6.4 package (Sueur et al. 2008) of the R statistical

environment, version 3.3.2 (R Core Team 2016).

Dominant frequency was estimated using a fast Fourier

transform and fundamental frequency was estimated

via short-term cepstral transform (Hanning window

length = 256 samples with 80% overlap between

successive windows for both spectral properties). 2D

spectrograms were visualized using a sliding window

analysis of short-term Fourier transform calculations.

Species descriptions. In order to simplify the

identification of the two new species, their

descriptions include comparisons of all the related

species. To avoid confusion as to the point at which

each species 1s formally described, the new species

names are followed by “sp. nov.” until the point in the

paper where a holotype is designated. Furthermore,

for the sake of clarity the epithet “sp. nov.” is also

used in the tables, figures, maps, appendices, and the

molecular tree. The epithet “sp.” is used for the new

species which are not described herein.

Results

Eleutherodactylus maculabialis sp. nov.

Spot-lipped Trilling Frog, Rana trinadora de labios

manchados.

Figs. 1-2, 7A.

urn:|sid:zoobank.org:act:55184A 71-4558-4E49-B379-73A4 8641939C9

Holotype. MZFC 33312 (CIG 00921). Adult male (Fig.

1), 11.4 km S of Puerto de Gallo, Municipality of Atoyac

Amphib. Reptile Conserv.

de Alvarez (17.4672, -100.1916, datum = WGS84; 1,980

m asl), Guerrero, Mexico (Fig. 8A), collected on 15

July 2016 by Christoph I. Griinwald and Héctor Franz-

Chavez.

Paratypes (n = 13; Fig. 2). MZFC 33307-311, 33313-

314 (CIG 00916—920; 00922923), seven adult males,

collected at same locality and on same date as holotype;

MZFC 33315-316 (CIG 00940—941), two adult males,

1.0 km S of bridge between Nueva Delhi and El Paraiso,

Municipality of Atoyac de Alvarez (17.4150, -100.1924

datum = WGS84; 1,320 m asl), Guerrero, Mexico,

collected on 15 July 2016 by Christoph Griinwald and

Héctor Franz-Chavez; MZFC 33317-319 (CIG 00945-—

00947), two adult males, Yerba Santa, Municipality of

General Heliodoro Castillo (17.5202, -99.9639, 2,003 m

asl; datum = WGS84), Guerrero, Mexico collected on

16 July 2016 by Christoph Griinwald and Héctor Franz-

Chavez; MZFC 33321 (CIG 00949), one adult male,

8.5 km N of Yerba Santa on road to Carrizal de Bravo,

Municipality of General Heliodoro Castillo (17.5269,

-99.9371, 1,880 m asl; datum = WGS84), Guerrero,

Mexico, collected on 16 July 2016 by Christoph

Grtinwald and Héctor Franz-Chavez.

Diagnosis. Based on the phylogenetic analysis, this

is a member of the genus Eleutherodactylus, subgenus

Syrrhophus, as defined by Hedges et al. (2008). It is in

the Eleutherodactylus (Syrrhophus) nitidus species series

and the Eleutherodactylus (Syrrhophus) nitidus species

group as defined by Grtinwald et al. (2018) based on

the condition of the tympanic annuli, ventral epidermis,

and visceral peritoneum. A small frog, adult males

measure 17.9—24.7 mm SVL; vocal slits are present in

males, readily visible under partially translucent ventral

epidermis; digital tips are expanded, 1.5—2.1 times the

width of the narrowest part the finger on the third and

fourth fingers; fingers moderately long, finger lengths

are I-]-IV-III with third finger length ranging from 17—

19% of SVL; compact lumbar gland above the inguinal

region present, indistinct, barely visible in live specimen;

ventral epidermis is partially translucent and visceral

peritoneum is clear, not white, thus abdominal vein is

not clearly visible against a white background on the

venter of live specimens, and viscera are partially visible

through translucent dark gray ventral epidermis; limbs

moderate, TL/SVL ratio is 0.44—0.50, FeL/SVL ratio

is 0.41-0.47 and TotFL/ SVL ratio is 0.66—0.73; snout

relatively short, END/ SVL ratio is 0.10-0.12; tympanum

small, indistinct and round, tympanic annuli not visible

in live specimen; TW/ED ratio is 0.25—0.29. The dorsal,

lateral, and ventral skin is smooth or slightly shagreen.

Dorsal coloration variable, orange, tan, or brown, with

darker brown marbling; loreal region dark brown, with

conspicuous large white or orange spots present on

upper labial region, and 1-3 orange spots on lower loreal

region and upper portion of upper labia; pale interorbital

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Grunwald et al.

Lt /

Fig. 1. Holotype of Eleutherodactylus maculabialis sp. nov., MZFC 33312 (CIG 00921) from 11.4 km S of Puerto de Gallo,

Municipio de Atoyac de Alvarez, Guerrero, Mexico. (A) Dorsolateral perspective in life. (B) Lateral perspective in life. (C) Ventral

perspective in life. (D) Dorsal perspective in preservative. (E) Ventral perspective in preservative.

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Two new Eleutherodactylus species from Mexico

Fig. 2. Some of the paratypes of Eleutherodactylus maculabialis sp. nov. in life. (A-—C) MZFC 33310 (CIG 00919); (D—F)

MZFC 33311 (CIG 00920); (G-Il) MZFC 33314 (CIG 00923) all from type locality; (J-L) MZFC 33321 (CIG 00949); (M-—O)

MZFC 33318 (CIG 00946); (P—R) MZFC 33317 (CIG 00945) all from the vicinity of Yerba Santa on road to Carrizal de Bravo,

Municipality of General Heliodoro Castillo, Guerrero, Mexico.

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Grunwald et al.

bar present, orange or tan, same color as palest dorsal

coloration; indistinct transverse bands present on legs;

upper arms same coloration and pattern as forearms;

no pale mid-dorsal stripe; upper flanks same color as

dorsum, lower flanks lavender or gray with white spots

and marbling; venter gray to dark gray with sparse

white spots or marbling present. Red, reddish, or orange

inguinal flash colors sometimes present on thighs and

groin; however, when present these colors are not more

vivid than the coloration of the light interorbital bar and

the characteristic colored spots on the upper lip. The

mating call of adult males is a short trill (see below;

Fig 3).

Comparisons. Eleutherodactylus maculabialis can be

distinguished from all species in the Eleutherodactylus

(Syrrhophus) longipes species series by: possessing a

small, indistinct tympanum with no tympanic annulus

visible and with a diameter less than 30% of diameter of

the eye; by possessing a ventral epidermis which is not

clear, and combined with a visceral peritoneum which

is not white, an abdominal vein on the venter which 1s

not clearly evident against a white background in life;

and by possessing a distinct, raised lumbar gland above

the inguinal region.

Eleutherodactylus maculabialis can be distinguished

from species of the Eleutherodactylus (Syrrhophus)

modestus species group by the combination of

possessing a compact, protruding lumbar gland above

the inguinal region, digital tips which are expanded

more than 1.5 times the width of the narrowest part the

finger on the third and fourth fingers, and the presence

of an interorbital bar.

Within its own species group, EF. maculabialis can

be distinguished from most species by possessing a

compact inguinal gland that is indistinct but visible

in live specimens. This character may or may not be

visible in preserved specimens depending on how they

were preserved. This species differs from E. pipilans, E.

erythrochomus, and E. nebulosus, which lack a visible

compact lumbar gland altogether. All other known

species in the E. (Syrrhophus) nitidus species group

have readily visible compact lumbar glands above the

inguinal region. Eleutherodactylus maculabialis can be

further distinguished from E. pipilans and E. nebulosus

by possessing digital tips which are expanded more

than 1.5 times the width of the narrowest part of the

finger, and from E. erythrochomus by possessing digital

tips which are more than 1.5 times but less than 3.0

times the width of the narrowest part of the finger. It

may be distinguished from E. nitidus, E. petersi, and

E. orarius by the combination of smoother skin, longer

limbs, and tips of digits which are expanded more than

1.5 times the narrowest part of the finger on the third

and fourth fingers. It is distinguished from E. albolabris

by the following characters (characters of E. albolabris

in parentheses): smaller size, 17.9—24.7 mm (23.0—26.8

mm), venter which is partially translucent and partially

Ampitute

Fig. 3. Oscillograms and spectrograms of the advertisement calls of Eleutherodactylus maculabialis sp. nov. and Eleutherodactylus

sentinelus sp. nov. (A) Adult male £. maculabialis sp. nov. from type locality. (B) Adult male E. maculabialis sp. nov. from

1.0 km S of bridge between Nueva Delhi and El Paraiso, Municipality of Atoyac de Alvarez, Guerrero, Mexico. (C) Adult male

Eleutherodactylus sentinelus sp. nov. from Puerto El Balsamo, Municipality of José Azueta, Guerrero, Mexico. (D) Adult male

Eleutherodactylus sentinelus sp. nov. from type locality.

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Two new Eleutherodactylus species from Mexico

white with black markings (entire venter white with

black markings), pale longitudinal blotch absent in

mid-dorsal area (pale mid-dorsal blotch usually present

in mid-dorsal area), smaller tympanum with TW/ED

0.25—0.28 (larger tympanum with TW/ED 0.27-0.32).

Eleutherodactylus maculabialis may be distinguished

from £. dilatus by the following characters (characters

of E. dilatus in parentheses): smooth dorsal and

ventral skin (rugose), dark gray ventral coloration

with white spots (white ventral coloration with dark

gray marbling), conspicuous white or orange spots

on labial region (dark labial region, sometimes with

white or pale yellow speckling), flash color absent,

or when present not brighter than the pale coloration

on dorsum and head (bright yellow flash coloration in

inguinal region). Eleutherodactylus maculabialis may

be distinguished from E. maurus by the following

characters (characters of E. maurus in parentheses):

presence of pale interorbital bar same color as snout

(no pale interorbital bar), dark gray ventral coloration

with white spots (white ventral coloration with dark

gray marbling), conspicuous white or orange spots on

labial region (dark labial region, sometimes with white

or pale yellow speckling), smooth dorsal and ventral

skin (rugose or slightly rugose dorsal and ventral skin),

no pale mid-dorsal stripe present and upper arms same

color as rest of limbs (thick, pale brown mid-dorsal

strip usually present and upper arms pale ground

coloration, unmarked and lighter than other limbs).

Eleutherodactylus maculabialis is most similar to E.

syristes. Both species share smooth dorsal and ventral

skin, distinct compact lumbar glands above the inguinal

region which are variably marked, a variable dorsal

coloration of orange, brown, or tan with darker marbling,

and similar red, reddish, or orange flash colorations.

Eleutherodactylus maculabialis may be distinguished

from E. syristes by the following characters (characters

of E. syristes in parentheses): digital tips expanded

more than 1.5 times the width of the narrowest part the

finger on the third and fourth fingers (digital tips which

are expanded 1.1—1.5 times the width of the narrowest

part the finger on the third and fourth fingers), ventral

coloration dark, with pale orange or white spots (ventral

coloration white, with dark gray or black marbling and

reticulations), conspicuous white or orange spots on

labial region (labial region dark, occasionally with light

gray or white speckling). Furthermore, EF. maculabialis

differs from all species in its species group by its

advertisement call, which is a short trill. The only other

species 1n the species group which has an advertisement

call that is a trill is &. syristes, but the call of that species

is much more drawn-out (see below, Fig. 11A—B).

Description of the holotype. Male, relatively small

(23.5 mm SVL); head longer (7.8 mm) than wide (6.9

mm), head slightly wider than body; snout subovoid

from a dorsal view and rounded from a lateral profile;

Amphib. Reptile Conserv.

tympanum indistinct, rounded with no supra-tympanic

fold present; tympanum small, circular, greatest width

of tympanum 0.7 mm; greatest diameter of eye 2.7 mm;

tympanum width to eye diameter 0.27; eyelid width 1.5

mm, approximately 32% of the IOD, first finger shorter

than second finger; finger lengths from shortest to

longest I-II-IV-IHI with two and three equal; digital pads

on fingers two, three, and four expanded, 1.9 times the

narrowest point of the digit on fingers three and four;

expanded finger pads slightly rounded, three palmar

tubercles; inner palmar tubercle about 70% as large as

middle palmar tubercle, outer palmar tubercle about

45% the size of middle palmar tubercle (Fig. 6A); toe

lengths from shortest to longest I-V-I-III-IV, TL2 and

TL5 very similar; outer metatarsal conical with a round

base, moderate, approximately 66% of inner metatarsal

tubercle; inner metatarsal tubercle spherical shape with

oval base, large, approximately 0.9 mm in length. Dorsal

skin smooth, lateral skin slightly shagreened, ventral

skin smooth to slightly areolate. Skin was smooth in

life. Vocal slits present.

In life the holotype had a reddish-tan dorsal

coloration on the back (Fig. 1A—C), with darker brown

mottling that was increasingly dense towards the mid-

dorsal area. The pale orange was present on flanks,

interorbital bar and spots on labial region. The flanks

were reddish-tan with indistinct darker brown mottling

and small white spots. The hind legs and arms were

reddish-tan with irregular brown transverse bars. Bright

reddish-orange flash colors on groin or thighs. Ventral

coloration was gray with white and orange spots on

throat and upper chest.

Coloration in preservative is light brown dorsum,

with darker brown markings and a pale tan interorbital

bar. The dorsal surfaces of the legs are light brown and

the groin and posterior surfaces of the thighs are brown.

The venter is white, with light brown on throat (Fig.

1D-E).

Measurements of the holotype (in mm). IND 2.5, IOD

4.8, END 2.5, ETD 0.9, UpL 6.3, FoL 7.5, HaL 5.5, FIL

1.8, FIPW 0.5, FLW 0.4, F2L 2.4, F2PW 0.7, F2W 0.4,

F3L 3.9, F3PW 0.9, F3W 0.5, F4L 2.4, F4APW 0.8, FAW

0.4, IPTL 0.7, MPTL 1.0, OPTL 0.4, FeL 10.5, TL 11.5,

TaL 5.9, FL 10.2, T2L 3.5, T2PW 0.8, T2W 0.5, T3L

4.6, T3PW 0.8, T3W 0.5, T4L 7.2, T4PW 0.9, T4W 0.5,

TSL 3.1, TSPW 0.6, TSW 0.4, IMTL 0.9, OMTL 0.6,

FeL/SVL 45%, TL/SVL 49%, Ha/SVL 23%, FL/SVL

43%, HL/SVL 33%, HW/SVL 29%.

Variation. SVL from 17.9-24.7 mm (21.65 + 1.77).

Expanded finger pads vary from 1.5—2.1 times the

narrowest part of the digit on the third and fourth

fingers, with average of 1.7 + 0.21 on the third finger

and average of 1.73 + 0.21 on the fourth finger. Dorsal

ground coloration ranged from different shades of tan,

reddish, or brown, always with darker brown coloration

February 2021 | Volume 15 | Number 1 | e272

Grunwald et al.

on the dorsum. Different degrees of dark mottling are

present on the dorsal surfaces. Venter always gray, but

with varying amounts of white spots. Characteristic

bright orange spots on upper part of upper labial and

lower part of loreal region present in most individuals

(MZFC 33307-314); however, these spots may be

faded or white in individuals from lower elevations

(MZFC 33315-19, 33321), and one specimen (MZFC

33316) has only one pale spot on the right side, and

none on the left. In some individuals, orange coloration

may be lacking on the lip (MZFC 33315-19, 33321),

in these individuals it is lacking everywhere, including

the inguinal region and on the interorbital bar.

Morphological variation is presented in Table 1.

Distribution and ecology. This species is known from

two municipalities in the Sierra Madre del Sur of central

Guerrero (Fig. 7A). It has been collected in tropical

evergreen forest, cloud forest, oak woodland, pine-oak

forest, and riparian vegetation between 900—2,100 m

asl. All specimens were collected from low-growing

vegetation or rocks along road cuts while calling in

the month of July. This species has been collected in

sympatry with E. albolabris and E. nitidus.

Etymology. From the combination of the Latin words

maculatus and labialis, meaning spotted-lip, named

for the conspicuous bright colored spots on the upper

lip and above the rictus, present in varying degrees,

which along with the advertisement call help to readily

distinguish this species in the field.

Referred specimens. CIG 01484—01485, 01501, three

adult males collected at the type locality on 29 June

2019 by Christoph I. Grinwald, Karen I. Morales-

Flores, Janelle Morales-Flores; JAC 22216, one adult

male, collected between Nueva Delhi and Puerto del

Gallo, Municipality of Atoyac de Alvarez (17.4668,

100.198, 2,020 m asl, datum WGS84), Guerrero,

Mexico on 15 June 2002 by Jonathan A. Campbell;

JAC 25643-25646, four adult males, collected at Nueva

Delhi, Municipality of Atoyac de Alvarez (17.4113,

-100.1954, 1,239 m asl, datum WGS84), Guerrero,

Mexico on 6 July 2004 by Jonathan A. Campbell.

Eleutherodactylus sentinelus sp. nov.

El Balsamo Peeping Frog, Rana piadora del Puerto El

Balsamo.

Figs. 4—5, 7B.

urn:lsid:zoobank.org:act: D89A77F0-2A F0-42D3-B69 B-DE90A CE8 FA 62

Holotype. MZFC 33306 (CIG 00913). Adult male (Fig.

4), 8.9 km SW of Puerto El Balsamo, Municipality of

José Azueta (17.9549, -101.2253, 1,354 m asl; datum =

WGS84), Guerrero, Mexico (Fig. 8B), collected on 14

July 2016 by Christoph I. Griinwald and Héctor Franz-

Chavez.

Amphib. Reptile Conserv.

Paratypes (n = 7; Fig. 5). MZFC 33302-33305 (CIG

00907-910), four adult males, collected at Puerto

El Balsamo, Municipality of José Azueta (17.9813,

-101.2291, 1,900 m asl; datum = WGS84), Guerrero,

Mexico on 14 July 2016 by Christoph I. Grinwald

and Héctor Franz-Chavez; MZFC 33031-33033 (CIG

00333-335), three adult males, collected at Puerto

El Balsamo, Municipality of José Azueta (17.9812,

-101.2292, 1,900 m asl; datum = WGS84), Guerrero,

Mexico on 5 June 2015 by Christoph I. Griinwald,

Nadia Pérez-Rivera, and Héctor Franz-Chavez.

Diagnosis. Based on the phylogenetic analysis, this

species is a member of the genus Eleutherodactylus,

subgenus Syrrhophus, as defined by Hedges et al. (2008);

and in the Eleutherodactylus (Syrrhophus) nitidus

species series and the Eleutherodactylus (Syrrhophus)

nitidus species group as defined by Grtinwald et al.

(2018) based on the condition of the tympanic annul,

ventral epidermis, and visceral peritoneum. A small

frog, adult males measure 23.3—25.3 mm SVL; vocal

slits are present in males; digital tips are expanded,

1.4—2.3 times the width of the narrowest part the finger

on the third and fourth fingers; finger lengths are I-II-

IV-III, fingers moderately long, with third finger length

ranging from 13-21% of SVL; compact lumbar gland

in the inguinal region present but indistinct, visible in

live specimen; ventral epidermis semi-translucent and

the visceral peritoneum is clear, not white, abdominal

vein barely visible on the venter of live specimens

against the background of the viscera; limbs moderate,

TL/SVL ratio is 0.41—0.56, FeL/SVL ratio is 0.38—0.46

and TotFL/ SVL ratio is 0.61—0.74; snout short, END/

SVL ratio is 0.10-0.11; tympanum small, indistinct and

round, tympanic annuli not visible in live specimen;

TW/EW ratio is 0.26—0.28. The dorsal skin is smooth

to slightly pustulate. Dorsal coloration is reddish-tan or

brown. A pale brown or reddish interorbital bar always

present, and a pale mid-dorsal blotch of the same color

as the interorbital bar is present. Upper arms pale and

unmarked, dark transverse bands present on forearms

and legs, and inguinal flash coloration orange or yellow

present on groin and sometimes on posterior portion of

thighs. Ventral coloration whitish or gray with some

darker gray spots or indistinct marbling. The mating

call of adult males is a quick chirp (“peep,” see below;

Fig. 3).

Comparisons. Eleutherodactylus sentinelus can be

distinguished from all species in the Eleutherodactylus

(Syrrhophus) longipes species series by: possessing a

small, indistinct tympanum with no tympanic annulus

visible and with a diameter less than 30% of diameter

of the eye; by possessing a visceral peritoneum which

is not white, so that the abdominal vein on the venter is

not clearly evident against a white background in life;

and by possessing a distinct, raised lumbar gland above

February 2021 | Volume 15 | Number 1 | e272

Two new Eleutherodactylus species from Mexico

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February 2021 | Volume 15 | Number 1 | e272

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Amphib. Reptile Conserv.

Grunwald et al.

Fig. 4. Holotype of Eleutherodactylus sentinelus sp. nov., MZFC 33306 (CIG 00913) from 8.9 km SW of Puerto El Balsamo,

Municipality of José Azueta, Guerrero, Mexico. (A) Dorsolateral perspective in life. (B) Lateral perspective in life. (C) Ventral

perspective in life. (D) Dorsal perspective in preservative. (E) Ventral perspective in preservative.

Amphib. Reptile Conserv. 11 February 2021 | Volume 15 | Number 1 | e272

Two new Eleutherodactylus species from Mexico

Fig. 5. Some of the paratypes of E/eutherodactylus sentinelus sp. nov. in life. (A-C) MZFC 33305 (CIG 00910); (D-F) MZFC

33304 (CIG 00909); (G—IT) MZFC 33032 (CIG 00334); (J-L) MZFC 33031 (CIG 00333); (M—O) MZFC 33033 (CIG 00335) from

Puerto El Balsamo, Municipality of José Azueta, Guerrero, Mexico.

Amphib. Reptile Conserv. 12 February 2021 | Volume 15 | Number 1 | e272

Grunwald et al.

the inguinal region.

Eleutherodactylus sentinelus can be distinguished

from species of the Eleutherodactylus (Syrrhophus)

modestus species group by the combination of

possessing a compact, protruding lumbar gland above

the inguinal region, digital tips which are expanded

more than 1.4 times the width of the narrowest part the

finger on the third and fourth fingers, and the presence

of an interorbital bar.

Within its own species group, E. sentinelus can

be distinguished from most species by possessing a

compact inguinal gland which is indistinct but visible

in live specimens. This differs from EF. pipilans, E.

erythrochomus, and E. nebulosus, which lack a distinct

visible inguinal gland altogether. All other known

species in the E. (Syrrhophus) nitidus species group have

readily visible compact lumbar glands above the inguinal

region. Eleutherodactylus sentinelus can be further

distinguished from E. pipilans, E. erythrochomus, and

E. nebulosus by the presence of distinct pale interorbital

bar and pale-yellow inguinal flash coloration. It may be

distinguished from E. nitidus, E. petersi, and E. orarius

by the combination of smoother skin, longer limbs,

and tips of digits which are expanded more than 1.4

times the narrowest part of the finger on the third and

fourth fingers. It further differs from these three species

by call, which is a short chirp rather than a whistle. It

is distinguished from E. albolabris by the following

characters (characters of FE. albolabris in parentheses):

lip dark, never white, with some flecking (lip white,

immaculate), inguinal flash coloration always yellow

(always fiery orange), ventral coloration translucent and

white, with some black markings (ventral coloration

completely white with bold black markings), tympanum

small with TW/ED 0.25-0.28 (tympanum slightly

larger with TW/ED 0.27-0.32). Eleutherodactylus

sentinelus may be distinguished from E. maurus by the

following (characters of £. maurus in parentheses): by

the presence of pale interorbital bar same color as snout

(E. maurus presents no pale interorbital bar), smooth

dorsal and ventral skin (rugose or slightly rugose dorsal

and ventral skin), bright yellow or orange flash colors

on thighs (flash colors absent, or barely discernible),

pale interorbital stripe distinct between darker head

coloration (pale interorbital stripe absent or same

color as dorsal surface of snout). Eleutherodactylus

sentinelus may be distinguished from FE. syristes and

E. maculabialis by its mating call, which is a rapid

chirp (“peep”) as opposed to a trill. Eleutherodactylus

sentinelus can be further distinguished from the closely

related E. syristes by having more expanded finger

tips, 1.4-2.3 times the narrowest part of the digit on

fingers three and four, and usually more than 1.6 (vs.

1.1-1.5 times the narrowest part of the digit on fingers

three and four), and by having yellow or yellowish-

orange flash colors (vs. orange or reddish flash colors).

Eleutherodactylus sentinelus is most similar to E.

Amphib. Reptile Conserv.

dilatus, from which it may be distinguished by the by

much smoother skin on both the dorsum and venter (E.

dilatus has rugose dorsal skin and pustulate venter),

smaller, less distinct inguinal glands (E. dilatus has

large, distinct inguinal glands), and a paler dorsal

ground coloration (E. dilatus is dark brown).

Description of the holotype. Small frog (24.1 mm

SVL); male; head slightly longer (7.1 mm) than wide

(6.4 mm), head slightly wider than body; snout subovoid

from a dorsal view and rounded from a lateral profile;

tympanum indistinct, rounded with no supra-tympanic

fold present; tympanum small, circular, greatest width

of tympanum 0.8 mm; greatest diameter of eye 2.9 mm;

tympanum width to eye diameter ratio 0.28; eyelid

width 1.6 mm, a third of the IOD; first finger shorter

than second finger; finger lengths from shortest to

longest I-II-IV-HI; digital pads on fingers two, three,

and four slightly expanded, 1.6 times the narrowest

point of the digit on second finger and 2.3 times the

narrowest point of the digit on fingers three and four;

expanded finger pads truncate; three palmar tubercles;

inner palmar tubercle about 75% as large as middle

palmar tubercle, outer palmar tubercle about half the

size of middle palmar tubercle (Fig. 6B); toe lengths

from shortest to longest I-H-V-II-IV; outer metatarsal

conical with a round base, small, approximately 50%

of inner metatarsal tubercle; inner metatarsal tubercle

spherical shape with oval base, large, approximately 0.9

mm in length; dorsal skin smooth, lateral skin slightly

shagreened, ventral skin smooth. Vocal slits present.

In life, the holotype had a dark reddish-brown dorsal

ground coloration, with pale reddish interorbital bar,

mid-dorsal blotch, and upper arm coloration, with the

upper arm the palest. Lateral portions of the head were

dark brown coloration, with small white flecks on the

labial and loreal regions. Lateral coloration was brown

and white. Legs and arms were ochre with dark brown

transverse bars. Yellow flash colors present on the groin.

Ventral coloration flesh colored with white spots and

black melanophores. See Fig. 4A—C for photographs of

the holotype in life.

Coloration in preservative is brown dorsum, with

some paler brown areas on the lower parts of the

back. Indistinct cream-colored interorbital bar, upper

arms and groin pale tan. Inguinal gland black. White

marbling on lateral surfaces. Legs and arms pale tan

with darker brown transverse bands. Ventral surfaces

white, unmarked, throat yellowish (Fig. 4D-E).

Measurements of the holotype (in mm). IND 2.4, IOD

4.8, END 2.5, ETD 1.1, UpL 6.2, FoL 7.9, HaL 6.3, FIL

2.3, FIPW 0.5, FIW 0.3, F2L 2.7, F2PW 0.6, F2W 0.4,

F3L 4.0, F3PW 1.0, F3W 0.4, F4L 3.1, F4PW 1.0, FAW

0.4, IPTL 0.6, MPTL 0.9, OPTL 0.4, FeL 11.3, TL 12.3,

TaL 7.2; FL 10.9, T2L 3.1, T2PW 0.7, T2W 0.4, T3L

4.9, T3PW 0.7, T3W 0.4, T4L 7.9, T4PW 0.8, T4W 0.4,

February 2021 | Volume 15 | Number 1 | e272

Two new Eleutherodactylus species from Mexico

Fig. 6. (A) Ventral aspect of hand of holotype of

Eleutherodactylus maculabialis sp. nov., MZFC 33312 (CIG

00921) from 11.4 km S of Puerto de Gallo, Municipio de Atoyac

de Alvarez, Guerrero, Mexico. (B) Ventral aspect of hand of

holotype of Eleutherodactylus sentinelus sp. nov., MZFC

33306 (CIG 00913) from 8.9 km SW of Puerto El Balsamo,

Municipality of José Azueta, Guerrero, Mexico.

TSL 3.5, TSPW 0.5, TSW 0.4, IMTL 1.0, OMTL 0.6,

FeL/SVL 46%, TL/SVL 50%, Ha/SVL 25%, FL/SVL

44%, HL/SVL 33%, HW/SVL 29%.

Variation. SVL from 23.3—25.3 mm (24.2 + 0.81).

Expanded finger pads on third and fourth fingers

vary from 1.4—2.3 times the narrowest part of the

digit, with average 1.85 + 0.26 on the third finger

and average 1.78 + 0.36 on the fourth finger. Dorsal

ground coloration ranges across different shades of

reddish brown (MZFC 33304), tan (MZFC 33031,

33306), and brown (MZFC 33032-3, 33304—5).

Venter typically gray, with white and black markings,

although these markings range from sparse to almost

complete reticulation. Morphological variation is

presented in Table 2.

Distribution and ecology. This species is known

only from the vicinity of the type locality in the

western-most extension of the Sierra Madre del Sur of

Guerrero (Fig. 7A). It has been collected at elevations

ranging from 1,300—1,900 m asl, on steep mountain

sides in humid pine-oak forest, oak woodland and

pine-oak woodland, and tropical deciduous forest

ecotone. At the type locality, this species 1s sympatric

with FE. petersi. All individuals of E. sentinelus have

been observed after the onset of the rainy season in

the months of June and July. Individuals were found

calling on small bushes or rocks. All were found

active at night.

Etymology. This species is named after latin sentinel,

meaning guard or outpost, in reference to its type

locality, a mountain which stands out from the north

and west as the first outpost of the Sierra Madre del

Sur of Guerrero.

Summary Data for Eleutherodactylus nitidus

Species Group Members

To help differentiate between the two new species

Amphib. Reptile Conserv.

described above, and the related species in the

Eleutherodactylus nitidus species group, important

morphological, mensural, and call differences of all

of the species in this group are collated into a single

table (Table 3), and photographs of related species in

the E. nitidus species group are provided (Figs. 9-10).

The distributions of the species in the E. nitidus group

are mapped in Figs. 7 and 13.

Differentiating the Advertisement Calls of

Closely Related Eleutherodactylus nitidus

Species Group Members

Recordings were made from 120 calling males of all

24 species of Eleutherodactylus from western Mexico.

The calls of the species analyzed were found to fall

into five different categories of a rapid burst whistle

(trill), a drawn-out whistle (whistle), a strong high-

pitched chirp (peep), a soft high-pitched chirp (chirp),

and a drawn-out chirp (pipe). Of the species analyzed

herein, four (E. dilatus, E. nebulosus, E. pipilans,

E. sentinelus) produce a strongly high-pitched chirp

(peep), four (E. albolabris, E. nitidus, E. orarius,

Eleutherodactylus related to E. nitidus

in southern Mexico

@ E sentinelus sp. nov.

@ £ maculabialis sp. nov.

O E dilatus

OE fuscus

OE syristes

Fleutherodactylus related to

E. pipilans in southern Mexico

@ E pipilans

®@® © nebulosus

Fig. 7. (A) Map showing the type localities and distribution

of Eleutherodactylus species related to E. nitidus in southern

Mexico. The stars represent type localities and circles represent

additional localities, with each color coded for the species: EF.

sentinelus sp. nov. (purple), £. maculabialis sp. nov. (green),

E. dilatus (pink), E. maurus (yellow), and E. syristes (blue).

(B) Map showing the type localities and distribution of

Eleutherodactylus species related to E. pipilans in southern

Mexico. The red star represents the type locality of E. pipilans

and red circles represent additional localities. The black star

represents the type locality of E. nebulosus and black circles

represent additional localities.

February 2021 | Volume 15 | Number 1 | e272

Grunwald et al.

il oF

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Fig. 8. (A) Type locality of Eleutherodactylus maculabialis

sp. nov. at 11.4 km S of Puerto de Gallo, Municipio de

Atoyac de Alvarez, Guerrero, Mexico. (B) Type locality of

Eleutherodactylus sentinelus sp. nov. at 8.9 km SW of Puerto

El] Balsamo, Municipality of José Azueta, Guerrero, Mexico.

E. petersi) produce a drawn-out whistle (whistle),

two (E. maculabialis and E. syristes) produce multi-

note or pulsed calls (trill) of various rates and notes,

and one (£. maurus) produces a slow and drawn-out

chirp (pipe). While a detailed account is given of the

calls of the species we describe here, as well as some

of their relatives, we refrain from giving detailed

descriptions of the calls of E. nitidus, E. albolabris, E.

orarius, E. petersi, E. pipilans, E. nebulosus, and “E.

rubrimactulatus” pending a more detailed taxonomic

study of this species complex.

Eleutherodactylus dilatus. The advertisement call

of E. dilatus is a single high-pitched “chirp” best

described as a “peep” (Fig. 11C). Calls recorded at the

type locality consist of a rapid chirp, with an average

duration of 110 ms, and a dominant frequency ranging

between starting at 2,550 kHz and 2,780 kHz.

Eleutherodactylus maculabialis. The advertisement

call of E. maculabialis is a relatively slow multi-note

“trill” (Fig. 3). Calls recorded at the type locality

consist of slow six-note trills with a duration of

180-210 ms, and a dominant frequency starting at

2,600—2,900 kHz and rising to 3,050—3,200 kHz. The

call interval is about 40-55 ms (Fig. 3A). At a nearby

Amphib. Reptile Conserv.

locality at 1,320 m asl (Fig. 3B), calls were recorded

of two males which varied from the type locality in

consisting of five-note trills with an average duration

of 230 ms and a dominant frequency starting at 3,630

kHz and reaching 4,000 kHz (Table 4).

Eleutherodactylus maurus. The advertisement call

of EF. maurus is a single, drawn-out “chirp” best

described as a “pipe.” Calls recorded near Ocuilan, in

Estado de México, consisted of a single chirp with a

duration of 205 ms, and a dominant frequency starting

at 2,760 kHz and rising to 3,150 kHz. The amplitude

was strongest at the middle of a call (Fig. 11D). Ata

locality near Taxco, Guerrero, a call of E. maurus was

recorded which consisted of a single, drawn-out chirp.

However, the call was longer, averaging 275 ms and

the amplitude was distributed in three sub-pulses. This

indicates a further need to investigate this population.

Eleutherodactylus sentinelus. The advertisement call

of E. sentinelus is a single high-pitched “chirp” best

described as a “peep” (Fig. 3). Calls recorded at the

type locality consist of a rapid chirp, with an average

duration of 160 ms, and a dominant frequency starting

at 2,560 kHz and rising to 3,100 kHz (Fig. 3D). Calls

recorded above the type locality at 1,900 m asl (Fig.

3C) are slightly shorter and of higher frequency,

with an average duration of 120 ms, and a dominant

frequency starting at 2,700 kHz and finishing at 3,300

kHz (Table 4).

Eleutherodactylus syristes. The advertisement call

of E. syristes is a rapid but long multiple-pulse “trill”

(Fig. 11A—B). Calls recorded at two different localities

in Guerrero consisted of 49-60 pulses, about 10 ms

apart, with a total duration of the call averaging 500-

560 ms. The dominant frequency at Agua de Obispo,

Guerrero, was between 2,700-3,170 kHz, starting

lower, peaking, and then lowering again (Fig. 11A).

At a second locality, slightly lower than Agua de

Obispo, east of Highway 95, the calls tended to have

a dominant frequency ranging between 3,000—3,390

kHz (Fig. 11B).

Eleutherodactylus albolabris. The advertisement

call of E. albolabris is a single, multi-pulsed whistle,

with the call continuous amongst the pulses and with

a duration of approximately 150-250 ms (Fig. 11F).

Calls recorded at the type locality at Agua de Obispo

had a duration of 160 ms, and started at a dominant

frequency of 2,840 kHz rising to 3,015 kHz. Calls

varied between 7—10 continuous pulses. At a second

locality, near Vallecitos in the Municipality of José

Azueta, Guerrero, the calls had a duration of 240 ms,

and started at a dominant frequency of 2,740 kHz which

rose to 2,915 kHz. Calls started with a multi-pulsed

trill and then ended with several distinct pulses. This

population’s call is best described as a combination of

a “trill” with a “whistle” (Fig. 11E).

February 2021 | Volume 15 | Number 1 | e272

Two new Eleutherodactylus species from Mexico

Table 2. Morphological measurements of E/eutherodactylus sentinelus sp. nov. specimens. See text for definitions of measurement

acronyms.

CIG-333 CIG-334 CIG-335 CIG-907 CIG-908 CIG-909 CIG-910 CIG-913

SVL 24.37 24.48 25.30 25-1 2351 23.41 23.25 24.54

HL 8.58 8.70 8.67 7.88 71 7.15 iA 8.03

TW 0.74 0.76 0.80 0.76 0.74 0.74 0.74 0.81

ED 2.63 2.68 2.04 2.67 2d 2.8 2.73 2.85

EIW 1.55 1.54 1.6 1.58 1.53 1.54 1.54 1.6

IOD 4.80 4.83 4.87 4.73 4.54 4.5 4.5 48

IND 2.58 2.37 259 2.34 2.31 2.33 2.28 2.44

END 2.80 2.74 2.78 2259 2.4 2.41 2.38 aoe

ETD 1.03 1.00 1.01 1.03 0.97 0.95 0.96 1.07

UpL 5.76 6.26 6.40 4.52 S32. 5.4 5237; 6.2

FoL 6.74 7.43 7.56 7.15 6.8 6.93 6.87 7.92

HaL 5.5 5.4 6.2 5.25 Sut 31 5.05 6.25

F3P'W/F3W 1.70 1.80 1.80 2.2 Ey 1.7 1.6 20

F4PW/F4W 1.60 1.90 1.70 23 1.4 |e 155 vies,

FeL 10.26 10.58 11.00 9.62 9.35 9.26 9.3 11.28

TL 11.03 11.65 11.33 10.31 9.95 10.12 o9 12.33

TotFL 16.4 16.3 17.5 15.4 iEoRey | 1535 15.4 18.05

IPT 0.58 0.70 0.74 0.5 0.58 0.6 0.59 0.65

MPT 0.84 1.00 1.07 0.77 0.88 0.86 0.87 0.86

OPT 0.4 0.45 0.45 0.32 0.39 0.42 0.42 0.4

IMTL 0.50 0.58 0.46 0.52 0.47 0.46 0.47 0.64

OMTL 0.90 0.92 1.04 0.74 0.84 0.85 0.83 0.99

TW/ED 0.28 0.28 0.29 0.28 0.27 0.26 0.27 0.28

Molecular Analysis of the Eleutherodactylus

nitidus Species Group

The molecular phylogeny based on _ the

mitochondrial rRNA 16S recovered a well-supported

Eleutherodactylus nitidus species group (posterior

probability (pp) 1; Figs. 12 and S1), which 1s sister

to the E. modestus group as defined by Griinwald et

al. (2018). As with previous phylogenies of the group

based on 16S, some of the intermediate nodes in the

phylogeny are not well supported (pp < 0.5), and we

have collapsed all nodes below this value.

In the Eleutherodactylus nitidus species group, we

recovered two main clades. The first one 1s composed

of FE. pipilans, E. erythrochomus, and E. nebulosus,

while the second group includes all the other species.

In the second group, many of the intermediate nodes

have little to no support. However, individual species

do show strong support (pp = 1) in most cases,

including £. dilatus, E. sentinelus, E. maurus, and E.

syristes, while E. maculabialis has a posterior support

of 0.86.

All the remaining species in the group form

a second, well-supported clade (pp =0.99). In

this clade, we recovered an early split between

Amphib. Reptile Conserv.

Eleutherodactylus albolabris and the other species,

which include &. orarius, E. petersi, E. nitidus, and

a hitherto undescribed form from Jalisco and Nayarit

(Fig. 13). We refrain from describing the western-most

form at this time, as material from the type locality

of FE. petersi is unavailable, so the relationship of the

western form with £. petersi remains unclear.

We believe that a more detailed study of the taxa

related to E. nitidus is needed, when more thorough

sampling is available and employing more population-

level specific tools, such as genome-wide SNP data,

in order to better understand the patterns of gene-flow

and introgression.

Discussion

With the descriptions of the two species here, the

number of taxa in the subgenus Syrrhophus increases

to 37, with 35 in continental North America and two

in Cuba. Guerrero is one of the most speciose states,

with nine recognized species and several undescribed

taxa awaiting description (Griinwald, pers. obs.). All

members of the subgenus Syrrhophus recorded from

Guerrero thus far belong to the E/eutherodactylus

nitidus species group as defined by Griinwald et

February 2021 | Volume 15 | Number 1 | e272

Grunwald et al.

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February 2021 | Volume 15 | Number 1 | e272

Amphib. Reptile Conserv.

Two new Eleutherodactylus species from Mexico

Table 4. Advertisement call data of E/utherodactylus maculabialis sp. nov. and Eleutherodactylus sentinelus sp. nov.

E. maculabialis

E. sentinelus

Call type Trill Peep

Dominant frequency (kHz) 3.28 + 532.5 2.91 + 85.0

Call length (ms) 212.5 + 20.55 140 + 20.0

Call rate (m') 5.49 241.1 2.84+0.19

Call rise time (ms) 135 22125 1344.0

Pulse rate 5.5 + 0.25 k+O

Call interval 19.87 + 4.28 36.94 + 10.52

al. (2018). Based on our phylogenetic analysis, E.

dilatus, E. maurus, and E. syristes are closely related

and restricted to high-elevation moist mountainous

areas. The new taxa described herein are closely

related to these species, with E. maculabialis being

a sister taxon to EF. syristes and E. sentinelus being

a sister taxon to E. dilatus. They collectively have

an allopatric distribution which covers most of

Guerrero’s high mountains.

Based on collection locality information,

E. maculabialis has a known distribution of

approximately 150 km? in the Sierra Madre del Sur.

Widespread on the windward slopes of the great Cerro

Teotepec, it has been collected at several localities

along the road that ascends it, and also at some

localities in the central Sierra Madre del Sur north of

the Cerro Teotepec (e.g., La Guitarra, Yerba Santa).

It appears to be isolated from the nearest locality of

its closest relative, E. syristes (Agua de Obispo), by

the combination of the dry and tropical Rio Papagayo

Valley and the cooler pine forests of Omiltemi area

which is inhabited by £. dilatus. Eleutherodactylus

maculabialis seems to be separated from E. dilatus

primarily by habitat, with the former inhabiting humid

pine-oak forest, cloud forest, and tropical wet forest-

oak woodland ecotone, whereas the latter is restricted

to slightly drier pine forest and pine-oak woodland.

A specimen (MZFC 12930) which appears to be E.

maculabialis, was collected at the western extreme of

the Omiltemi State Park. Unfortunately, it is not well

preserved, and the absence of molecular data makes

it impossible to definitively assign this specimen to

this species. However, if this population is indeed BE.

maculabialis, it would suggest a much larger range

for this species and closer proximity to the range of

E. dilatus.

The limited number of known collecting localities

for E. sentinelus suggest that it has the smallest

range of the E. nitidus species group, seconded by

its close relative, E. dilatus. As currently understood,

the distribution of EF. sentinelus is approximately

22 km’, while £. dilatus has a known distribution

of approximately 76 km?. These two species appear

to be separated by around 160 km of Sierra Madre

del Sur where neither has been collected, and only

the lower elevation species of E. nitidus, E. petersi,

E. albolabris, and E. maculabialis have been

collected. More sampling is needed to determine the

relationship of the distributions of &. sentinelus and

Amphib. Reptile Conserv.

E. maculabialis in the western portions of the Sierra

Madre del Sur.

Eleutherodactylus maculabialis is a species that

seems to be abundant where it occurs. While exact

man-hours per specimen collected were not logged

while collecting this species, numerous specimens

were collected with each attempt to locate the species,

and many more were heard calling. E/eutherodactylus

sentinelus seems to be less abundant than its congeners.

During four attempts to collect specimens during

the breeding season (June and July), no specimens

were heard or collected during one of them, only

three specimens were heard during another, and the

remaining two attempts resulted in more than a dozen

specimens heard and collection of the type series.

This contrasts with our experiences with closely

related E. dilatus, E. maculabialis, and E. syristes, of

which dozens of specimens were heard calling at all

localities during all attempts to collect these species.

From a conservation perspective, E. maculabialis

does not appear to have any major threats at this time.

Small-scale agriculture is present at some locations

throughout its range, but does not seem to largely

impact this species, and the species continues to

be present in disturbed plots. However, both illegal

logging activity and illegal poppy farming were

previously rampant on the lower slopes of Cerro

Teotepec. In addition to destroying the forest being

logged directly, the illegal logging also causes heavy

siltation and landslides which destroy large swaths

of the surrounding vegetation. The illegal poppy

plantations also caused large swaths of suitable habitat

to be completely cleared. We propose this species to

be provisionally classified as Endangered Blab(iil),

based on IUCN Red List criteria that the range is more

than 100 km? but smaller than 5,000 km/?, it occurs in

only a couple of threat-defined locations, and there 1s

ongoing decline in the extent and quality of its habitat

due to small-scale agriculture, cattle ranching, illegal

logging, opium poppy farming, road construction, and

the resulting siltation due to these activities.

Habitat transformation due to _ small-scale

agriculture is widely present within the distribution

range of E. sentinelus, which seems to be limited to

approximately 22 km*. While these frogs seem to

persist in disturbed areas, they are much less abundant

than congeners in similar areas (Griinwald, pers. obs.,

see above). Due to the relative rarity of these frogs

combined with the small known distribution, we

February 2021 | Volume 15 | Number 1 | e272

Grunwald et al.

Fig. 9. Comparison photos of E/eutherodactylus species related to E. nitidus in life. (A—I) E. syristes from the vicinity of Agua de

Obispo, Municipality of Chilpancingo, Guerrero, Mexico. (J—O) E. albolabris from Agua de Obispo, Municipality of Chilpancingo,

Guerrero, Mexico. (P-R) E. nitidus from Yerba Santa, Municipality of General Heliodoro Castillo, Guerrero, Mexico.

Amphib. Reptile Conserv. 19 February 2021 | Volume 15 | Number 1 | e272

Two new Eleutherodactylus species from Mexico

Fig. 10. Comparison photos of Eleutherodactylus species related to E. nitidus in life. (A-D) E. dilatus from Omiltemi, Municipality of

Chilpancingo, Guerrero, Mexico. (J—O) E. petersi from Puerto El Balsamo, Municipality of José Azueta, Guerrero, Mexico. (P-R) E.

pipilans from Acahuizotla, Municipality of Chilpancingo, Guerrero, Mexico.

Amphib. Reptile Conserv. 20 February 2021 | Volume 15 | Number 1 | e272

Grunwald et al.

Ampdiude

Frequency [kHa}

aod

i ae

5 4 — a =

E ' stbdbhee

;

oz 63 ea

Time (5)

Frequency (kHz)

Acpliute

tam

Fig. 11. Oscillograms and spectrograms of the advertisement calls of adult males of Eleutherodactylus species related to E.

maculabialis sp. nov. and E. sentinelus sp. nov. (A) E. syristes from Agua del Obispo, Guerrero, Mexico. (B) E. syristes from east of

Hwy. 95, near Acahuizotla, Guerrero, Mexico. (C) E. dilatus from Municipality of Chilpancingo, Guerrero, Mexico. (D) E. maurus

from Municipality of Ocuilan, Estado de México, Mexico. (E) E. albolabris from Municipality of Agua de Obispo, Guerrero,

Mexico. (F) E. albolabris from Vallecitos, Guerrero, Mexico.

propose this species to be provisionally classified as

Critically Endangered Blab(iii), based on the IUCN

Red List criteria that its occurrence is less than 100

km7, it occurs in only one threat-defined location, and

there is ongoing decline in the extent and quality of its

habitat due to small-scale cattle ranching and maize

farming.

The species richness of the subgenus Syrrhophus

in Guerrero (9 species) represents 27% of the species

richness in Mexico (35 species). The number of species

known in this state will likely increase as more isolated

mountain regions of Guerrero are explored. Guerrero

currently has 83 species of amphibians, which

represents 20.3% of the approximately 407 species

currently recognized for Mexico (AmphibiaWeb

2020). More research in Guerrero is necessary to

Amphib. Reptile Conserv.

continue documenting the incredible biodiversity of

this state, as rampant habitat destruction and illegal

logging continue to destroy the unique amphibian

habitats in the forests of the Sierra Madre del Sur

(Lips et al. 2004).

On the Validity of Eleutherodactylus nebulosus

Taylor, 1943 and E. rubrimaculatus Smith and

Taylor, 1945 as Species

Based on our phylogenetic results and genetic

distances, the populations referred to as “E.

rubrimaculatus” appear to be conspecific with E.

nebulosus, as these two species are paraphyletic

with respect to each other. Additionally, the genetic

distances between the two taxa are very low (less than

February 2021 | Volume 15 | Number 1 | e272

Two new Eleutherodactylus species from Mexico

Other outgroups

- EU186711 E. erythrochomus

- JAC 25809 E. pipilans - Pochutla, OAX

0.63 [ggg me: SAC 25810 E. pipilans - Pochutla, OAX

; ap ROEes cee JAC 25811 E. pipilans - Pochutla, OAX

- CIG 440 E. pipilans - Malinaltepec, GRO

JAC 24283 E. pipilans Rio Grande, OAX

CIG 396 E. pipilans - Acahuizotla, GRO

- CIG 398 E. pipilans - Acahuizotla, GRO

CIG 755 E. rubrimaculatus - Huixtla, CHIS

- CIG 753 E. rubrimaculatus - Ujuarez, CHIS

CIG 1240 E. nebulosus - Ocozocoautla, CHIS

CIG 1241 E. nebulosus - Ocozocoautla, CHIS

0-98) F- CIG 1236 E. nebulosus - Trinitaria, CHIS

~ CIG 1237 E. nebulosus - Trinitaria, CHIS

- CIG 1238 E. nebulosus - Trinitaria, CHIS

- CIG 407 E. dilatus - Omiltemi, GRO

- CIG 408 E. dilatus - Omiltemi, GRO

CIG 333 E. sentinelus - Altamirano Hwy., GRO

CIG 334 E. sentinelus - Altamirano Hwy., GRO

CIG 335 E. sentinelus - Altamirano Hwy., GRO

CIG 380 E. maurus - Ocuilan, MEX

~ CIG 382 E. maurus - Ocuilan, MEX

- CIG 385 E. maurus - Huitzilac, MOR

CIG 387 E. maurus - Huitzilac, MOR

- CIG 388 E. maurus - Huitzilac, MOR

» JAC 25643 E. maculabialis - Paraiso - Nueva Dehli, GRO

CIG 940 E. maculabialis - Puerto de Gallo Road, GRO

* CIG 941 E. maculabialis - Puerto de Gallo Road, GRO

- JAC 25644 E. maculabialis - Paraiso-Nueva Dehli, GRO

- JAC 25645 E. maculabialis - Paraiso-Nueva Dehli, GRO

- JAC 25646 E. maculabialis - Paraiso-Nueva Dehli, GRO

CIG 921 E. maculabialis - Type Locality, GRO

CIG 923 E. maculabialis - Type Locality, GRO

CIG 949 E. maculabialis - Yerba Santa, GRO

CIG 946 E. maculabialis - Yerba Santa, GRO

CIG 947 E. maculabialis - Yerba Santa, GRO

- CIG 954 E. syristes - Agua del Obispo, GRO

- CIG 1233 E. syristes - Loxicha, OAX

- CIG 1232 E. syristes - Loxicha, OAX

- CIG 627 E. syristes - Type Locality, OAX

CIG 628 E. syristes - Type Locality, OAX

CIG 713 E. syristes - Oaxaca City, OAX

-~- CIG 714 E. syristes Sta. Ma. Yucuhiti, OAX

somone ANMO 2999 E. syristes - Malinaltepec, GRO

0.99) Pon JAC 25701 E. syristes - Malinaltepec, GRO

cree JAC 25702 E. syristes - Malinaltepec, GRO

E. erythrochomus

E. pipilans

0.74

E. nebulosus

E. dilatus

E. sentinelus sp. nov.

E. maurus

E. maculabialis sp. nov.

E. syristes

coven JAC 25703 E. syristes - Malinaltepec, GRO

veriepeereeeerte CIG 431 E. syristes - Malinaltepec, GRO

eee CIG 434 E. syristes - Malinaltepec, GRO

O.95b OIG 435 E. syristes - Malinaltepec, GRO

voowecenns GIG 391 E. albolabris - Agua del Obispo, GRO

~ CIG 392 E. albolabris - Agua del Obispo, GRO

- CIG 953 E. albolabris - Agua del Obispo, GRO

~ CIG 441 E. albolabris - Malinaltepec_GRO

- JAC 25586 E. albolabris - Atoyac-Nueva Dehli, GRO

E. albolabris

O73>a

nae ae CIG 331 E. albolabris - Altamirano Hwy., GRO

vomewerens CIG 332 E. albolabris - Altamirano Hwy., GRO

“Boks p00” CIG 330 E. albolabris - Altamirano Hwy., GRO

- JAC 25642 E. albolabris - Coyuquilla - Durazno, GRO

CIG 310 E. petersi - Sierra del Tigre, JAL

- CIG 311 E. petersi - Sierra del Tigre, JAL

» JAC 25219 E. petersi - Altamirano Hwy., GRO

~ JAC 25265 E. petersi- El Balsamo, GRO

- JAC 25266 E. petersi- El Balsamo, GRO

JAC 25299 E. petersi - Vallecitos, GRO

CIG 336 E. petersi - Altamirano Hwy., GRO

- CIG 337 E. petersi- Altamirano Hwy., GRO

CIG 314 E. sp. nov. - JAL

CIG 649 E. sp. nov. - Sta. Maria del Oro, NAY

CIG 412 E. nitidus - Omiltemi, GRO

CIG 439 E. nitidus - Mixtecapa, GRO

EU186712 E. nitidus

soe JAC 25815 E. nitidus - Marquelia - Tlapa, GRO

aa CIG 389 E. nitidus - Higueron, MOR

0.8 - CIG 715 E. nitidus - Sta. Ma. Yucuhiti - OAX

ari DQ283316 E. nitidus

sos JAC 24020 E. orarius Hwy. 200, MICH

JAC 25341 E. orarius - Lazaro Cardenas, MICH

JAC 25342 E. orarius - Lazaro Cardenas, MICH

JAC 25343 E. orarius - Lazaro Cardenas, MICH

~ JAC 25344 E. orarius - Lazaro Cardenas, MICH F

~ CIG 458 E. orarius - Ixtlahuacan, COL E. orarius

- CIG 460 E. orarius - Ixtlahuacan, COL

JAC 25563 E. orarius Hwy. 200, MICH

JAC 25564 E. orarius Hwy. 200, MICH

CIG 341 E. orarius - Aquila, MICH

» JAC 25526 E. orarius - Hwy. 200, MICH

0.99

E. petersi

E. sp.

0.84

0.77

E. nitidus

Fig. 12. Bayesian phylogenetic inference of members of the E/eutherodactylus subgenus Syrrhophus, with a focus on the E. nitidus

Species group, based on the mitochondrial loci 16S rRNA. Black circles represent nodes with a posterior support of 1. All nodes

with support of less than 0.5 are collapsed.

Amphib. Reptile Conserv. 22 February 2021 | Volume 15 | Number 1 | e272

Grunwald et al.

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February 2021 | Volume 15 | Number 1 | e272

Amphib. Reptile Conserv.

Two new Eleutherodactylus species from Mexico

Eleutherodactylus related to

E. nitidus in southern Mexico

@ E. sp.

@ E. orarius

@ E. albolabris

@ E nitidus

O E petersi

Fig. 13. Map of type localities and distributions of members

of the Eleutherodactylus nitidus species group. Purple circles

represent localities of an undescribed species related to E.

petersi. For the other four species, stars represent the type

localities and circles represent additional localities, which are

color coded for species: E. orarius (red), E. albolabris (blue),

E. nitidus (green), and E. petersi (orange).

1.5%; Table 5).

Taylor (1940) first described E. nebulosus from

Arriaga, Chiapas, in 1943; and Smith and Taylor

(1945) then described “E. rubrimaculatus” from

Finca La Esperanza, near Escuintla, Chiapas. We did

not collect this species at the type locality, however

(CIG 755) is near-topotypic, from less than 19 km

away on the same slope of the same mountain.

The authors differentiated between these species

predominately based on size and color pattern.

However, those characters appear to be clinal when

comparing photos of individual frogs from six

localities along Pacific Chiapas (Fig. 14)—from west

(Arriaga) to east (Union Juarez) this is evident in the

color pattern shifting from the E. nebulosus pattern

(Arriaga) to the “E. rubrimaculatus”’ pattern (Union

Juarez). While molecular material was not available

from all the individuals associated with these photos,

our analysis presented here did include molecular

material from two localities of “E. rubrimaculatus”

and several localities of EF. nebulosus around

Chiapas. While closely related species of Syrrhophus

occurring in sympatry or near sympatry normally

have very different advertisement calls, populations

of E. nebulosus at Arriaga, Chiapas, and populations

of “E. rubrimaculatus” at Huixtla, Chiapas, produce

the same single note “peep” (Griinwald, pers. obs.).

This suggests they are not reproductively isolated.

Furthermore, the genetic distances are very low.

(1.1-1.3%; Table 5). We thus conclude that “E.

rubrimaculatus” is most likely a junior synonym of

E. nebulosus. Our molecular analysis of frogs related

to E. pipilans \end support for the recognition of

two clades within E. pipilans, which are consistent

with E. pipilans from Guerrero and Oaxaca and E.

nebulosus + “E. rubrimaculatus” from southeastern

Oaxaca, Chiapas, and Guatemala (Fig. 7B). Herein we

recognize two species, EF. pipilans and E. nebulosus.

However, we stress that the lack of genetic sampling

Amphib. Reptile Conserv.

between south-central Oaxaca and northwestern

Chiapas may be exacerbating the actual genetic

distances between these two populations. We suspect

that further sampling around Cerro Quiengola,

Tehuantepec, and the Sierra Sacamecate may prove

these taxa to be two subspecies of a wide-ranging

species.

Common Names for Members of the

Eleutherodactylus nitidus Species Group

Although species of Syrrhophus are often difficult

to distinguish, their advertisement calls are useful

for distinguishing species in the field. As these

frogs are often located during breeding season by

following their advertisement calls, it is helpful to

know what type of call each species emits. Grinwald

et al. (2018) defined vernacular names for the E£.

modestus species group based on the nature of their

distinct advertisement calls. Similarly, we propose

common names for the species described herein, as

well as other species of the E. nitidus species group

in Appendix 3.

Acknowledgements.—We thank Nadia Pérez-Rivera,

Brandon T. La Forest, and André Joao Griinwald

for risking their lives in the sierra of Guerrero and

providing valuable assistance in the field. Dr. Vinicius

Guerra-Batista of the Centro de Ciéncias Bioldgicas

e da Natureza at the Universidade Federal do Acre,

Brazil, generously donated his time and shared his

expertise to help us analyze the advertisement calls

of various species of Eleutherodactylus. We are

indebted to the following persons for allowing us

to examine specimens or providing us with digital

photographs of preserved specimens under their care:

Dr. Adrian Nieto-Montes de Oca, Dr. Oscar Flores-

Villela, and Edmundo Pérez-Ramos at MZFC; and

Dr. Jonathan Campbell and Dr. Gregory Pandelis at

UTA. All specimens which were deposited in MZFC

were deposited under permit #FAUT-0093 issued to

Dr. Adrian Nieto Montes de Oca. Permits were issued

by the Secretaria de Media Ambiente y Recursos

Naturales (SEMARNAT). We especially thank Dr.

Adrian Nieto Montes de Oca and the Universidad

Nacional Autonoma de México, Museo de Zoologia

de la Facultad de Ciencias, for his generous support

in furthering our understanding of the Mexican

herpetofauna. We thank Tom J. Devitt and two

anonymous reviewers who provided extensive

comments, critique, and advice on an earlier version

of this manuscript. An extra-special thanks goes to

André, Ambar Lanomy, Emilio, Janelle, and Ximena

for sharing their parents’ enthusiasm and love of

nature, and for doing field work at such an early age!

Biencom Real Estate, Biodiversa A.C., and Herp.mx

A.C. provided important funding for the field work.

February 2021 | Volume 15 | Number 1 | e272

Grunwald et al.

r 5. 5 ae

Fig. 14. Eleutherodactylus nebulosus in life, including specimens formerly assigned to ‘“Eleutherodactylus rubrimaculatus.” (A)

E. nebulosus from the Municipality of Mapastepec, Chiapas. (D) “E. rubrimaculatus” (=E. nebulosus) from the Municipality of

Huixtla, Chiapas. (E) “E. rubrimaculatus” (=E. nebulosus) from Belisario Dominguez, Chiapas. (F) “E. rubrimaculatus” (=E.

nebulosus) from the Municipality of Union Juarez, Chiapas.

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Christoph I. Griinwald is aGerman-Mexican herpetologist whose specialty 1s conservation

through field research. Chris leads expeditions for the HERP.MX Field Team, which have

resulted in numerous range extensions and state records, 15 new species descriptions, and

the re-discovery of several “extinct” species. For years Chris specialized in rattlesnakes and

pitvipers, and while many important discoveries have involved these groups, the need for

greater conservation efforts has turned his attention to amphibians—many of which remain

profoundly understudied in Mexico. Currently, Chris leads research expeditions in southern

Mexico aimed at documenting the novel species in threatened environments, rediscovering

amphibians thought to be extinct, and collecting data for concerted conservation efforts.

As a co-founder of Biodiversa, A.C., an anti-extinction non-profit organization, Chris is

working to develop a system of “micro-reservas,” to help conserve the most vulnerable

high-endemism localities in Mexico.

Jacobo Reyes-Velasco is originally from Colima, Mexico, although he has worked

around the world. Jacobo received his Bachelor’s degree in Biology from the Universidad

de Guadalajara (CUCBA), and later received his Ph.D. from the University of Texas

at Arlington. He is currently leading field expeditions in Mexico, teaching molecular

biology, and writing a book on the herpetofauna of Colima. He has published numerous

papers on the herpetofauna of western Mexico and is the co-founder of Entorno

Biodtico A.C. and HERP.MX A.C., two NGOs that focus on conservation initiatives and

sustainable development in western Mexico.

Hector Franz-Chavez is a herpetologist and marine mammal specialist originally

from Guadalajara, México. Hector Franz is a student of biology at the University of

Guadalajara (CUCBA) and has been an avid herpetologist since childhood. His main

interests are biogeography, natural history, and ecology of the herpetofauna of Mexico.

He is also an avid nature photographer and marine tour guide in the Sea of Cortez, and has

collaborated on ten species descriptions as well as numerous herpetological inventories

of different parts of Mexico. Currently stationed out of La Paz, Baja California Sur,

Hector is working on photographing and documenting the herpetofauna and marine life

of the Baja California Peninsula.

Amphib. Reptile Conserv.

February 2021 | Volume 15 | Number 1 | e272

QLAys

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DE1 CARM! avi /) Ps

Amphib. Reptile Conserv.

Two new Eleutherodactylus species from Mexico

Karen I. Morales-Flores is a herpetologist originally from Guadalajara, Mexico, but

she grew up in the United States where she developed an appreciation for nature, and

specifically the reptiles in the deserts of Nevada. A student of biology at the University of

Guadalajara (CUCBA), Karen (or “Kim’’) has participated in numerous field expeditions

with the HERP.MX Field Team. Her skills in the laboratory have encouraged her to take

on ambitious projects like measuring every external body part of 500 direct-developing

frogs. Her research has resulted in the description of nine new species so far, with many

more to come. Kim’s ambitions as a herpetologist are only matched by her ambitions

as an explorer, and she is currently travelling the world looking for exciting research

opportunities.

Ivan T. Ahumada-Carrillo is a Mexican herpetologist from Guadalajara, who received

his degree from the University of Guadalajara (CUCBA). Ivan is currently an independent

investigator who focuses on the biogeography of reptiles and amphibians in western

Mexico. He has documented dozens of range extensions and state records, and has

authored and co-authored various papers on biogeography, as well as the book Anfibios y

Reptiles del Bosque La Primavera. Another interest of his 1s wildlife photography, and his

work has been published throughout Mexico in educational materials, web sites, scientific

magazines, and books. Ivan has now co-authored 11 new species descriptions, as well as

numerous range extensions and state records from Mexico.

Christopher M. Rodriguez is a herpetologist from Los Angeles, California, who studied

biology at California State University. Chris is an avid breeder of endangered reptiles and

amphibians. Working at the Los Angeles Zoo, Chris has led captive breeding projects

for endangered herpetofauna from around the world, as well as extremely delicate local

species such as the Mountain Yellow-legged Frog (Rana muscosa). When not saving

species, Chris has led field expeditions around the world, including Madagascar, Indonesia,

Thailand, and above all Mexico. As a member of the HERPMX Field Team, Chris has

helped with the initial field work for several of the new species the Team has discovered.

Jason M. Jones was born and raised in southern California, where he studied biology and

computer science at the University of California, Irvine. Currently residing in Colima,

he has spent the past 17 years studying and photographing the herpetofauna of Mexico.

A specialist in Mexican pitvipers, Jason co-led the rediscovery of Crotalus lannomi,

and collaborated in the recent descriptions of Crotalus campbelli, Crotalus tlaloci, and

Ophryacus smaragdinus. Jason co-founded HERP.MX which, in collaboration with

Biodiversa, A.C., is currently developing strategies for the conservation of at-risk reptile

species.

28 February 2021 | Volume 15 | Number 1 | e272

Grunwald et al.

172585_E_zeus_Cuba

~ EF493718_E_ zeus

- 172583_E_symingtoni_Cuba

EF493643_E_symingtoni

CIG_611_E_longipes__Santiago_NL

CIG_619_E_smithi_Realde14_SLP

- CIG_813_E_verrucipes_Miquihuana_TAM

CIG_1270_E_leprus_Chimalapas_VER

CIG_1142_E_leprus_Zongolica_VER

~ CIG_1144_E_leprus_Zongolica_VER

- CIG_893_E_manantlanensis_Terrero_COL

CIG_544_E_rufescens_Tapalpa_JAL

CIG_563_E_rufescens_Coalcoman_MICH

CIG_857_E_modestus_TypeLocality_COL

JRV_160_E_wixarika_Sierra_Huichola_JAL

CIG_984_E_nietoi_Cave_MICH

~ JRV_142_E_grunwaldi_Toxin_JAL

- EU186711_E_pipilans

- JAC_25809_E_pipilans_Hwy175_Pochutla_OAX

~ JAC_25810_E_pipilans_Hwy175_Pochutla_OAX

~ JAC_25811_E_pipilans_Hwy175_Pochutla_OAX

CIG_440_E_pipilans_Malinaltepec_GRO

JAC_24283_E_pipilans_RioGrande_OAX

- CIG_396_E_pipilans_Acahuizotla_GRO

CIG_398_E_pipilans_Acahuizotla_GRO

CIG_755_E_rubrimaculatus_Huixtla_Chis

~ CIG_753_E_rubrimaculatus_Ujuarez_CHIS

- CIG_1240_E_nebulosus_Ocozocoautla_CHIS

CIG_1241_E_nebulosus_Ocozocoautla_CHIS

- CIG_1236_E_nebulosus_Trinitaria_CHIS

~4 CIG_1237_E_nebulosus_Trinitaria_CHIS

- CIG_1238_E_nebulosus_Trinitaria_CHIS

CIG_407_E_dilatus_Omiltemi_GRO

- CIG_408_E_dilatus_Omiltemi_GRO

~ CIG_333_E_sentinelus_Altamirano_Hwy_GRO

~ CIG_334_E_sentinelus_Altamirano_Hwy_GRO

~ CIG_335_E_sentinelus_Altamirano_Hwy_GRO

- CIG_380_E_maurus_Ocuilan_MEX

CIG_382_E_maurus_Ocuilan_MEX

CIG_385_E_maurus_Huitzilac_MOR

- CIG_387_E_maurus_Huitzilac_MOR

CIG_388_E_maurus_Huitzilac_MOR

JAC_25643_E_maculabialis_ParaisoNuevaDehli_GRO

CIG_940_E_maculabialis_PdGRoad_low_GRO

CIG_941_E_maculabialis_PdGRoad_low_GRO

JAC_25644_E_maculabialis_ParaisoNuevaDehli_GRO

JAC_25645_E_maculabialis_ParaisoNuevaDehli_GRO

* JAC_25646_E_maculabialis_ParaisoNuevaDehli_GRO

CIG_921_E_maculabialis_TypeLoc_GRO

CIG_923_E_maculabialis_TypeLoc_GRO

CIG_949_E_maculabialis_N_YerbaSanta_GRO

CIG_946_E_maculabialis_YerbaSanta_GRO

- CIG_947_E_maculabialis_YerbaSanta_GRO

- CIG_954_E_syristes_AguadeObispo_GRO

- CIG_1233_E_syristes_Loxicha_OAX

CIG_1232_E_syristes_Loxicha_OAX

CIG_627_E_syristes_TypeLoc_OAX

- CIG_628_E_syristes_TypeLoc_OAX

CIG_713_E_syristes_OaxacaCity_OAX

CIG_714_E_syristes_StaMaYucuhiti_OAX

- ANMO_2999_E_syristes_Malinaltepec_GRO

- JAC_25701_E_syristes_Malinaltepec_GRO

JAC_25702_E_syristes_Malinaltepec_GRO

JAC_25703_E_syristes_Malinaltepec_GRO

- CIG_431_E_syristes_Malinaltepec_GRO

CIG_434_E_syristes_Malinaltepec_GRO

CIG_435_E_syristes_Malinaltepec_GRO

- CIG_391_E_albolabris_AguadeObispo_GRO

- CIG_392_E_albolabris_AguadeObispo_GRO

- CIG_953_E_albolabris_Type_AguadeObispo_GRO

- CIG_441_E_albolabris_Malinaltepec_GRO

- JAC_25586_E_albolabris_AtoyacNuevoDehli_GRO

CIG_331_E_albolabris_Altamirano_Hwy_GRO

CIG_332_E_albolabris_Altamirano_Hwy_GRO

- CIG_330_E_albolabris_Altamirano_Hwy_GRO

JAC_25642_E_albolabris_CoyuquillaDurazno_GRO

CIG_310_E_petersi_Sierra_del_Tigre_JAL

~ CIG_311_E_petersi_Sierra_del_Tigre_JAL

- JAC_25219_E_petersi_AltamiranoHwy_GRO

JAC_25265_E_petersi_ElIBalsamo_GRO

JAC_25266_E_petersi_EIBalsamo_GRO

- JAC_25299 E_petersi_Vallecitos_GRO

CIG_336_E_petersi_Altamirano_Hwy_GRO

CIG_337_E_petersi_Altamirano_Hwy_GRO

- CIG_314_E_spnov_JAL

~ CIG_649_E_spnov_StaMariadelOro_NAY

~ CIG_412_E_nitidus_Omiltemi_GRO

~ CIG_439_E_nitidus_Mixtecapa_GRO

- EU186712_E_nitidus

JAC_25815_E_nitidus_Marquelia_Tlapa_GUER

CIG_389_E_nitidus__Higueron_MOR

- CIG_715_E_nitidus_StaMaYucuhiti_OAX

DQ283316_E_nitidus

JAC_24020_E_orarius_Hwy200_MICH

~ JAC_25341_E_orarius_LazaroCard_MICH

- JAC_25342_E_orarius_LazaroCard_MICH

JAC_25343_E_orarius_LazaroCard_MICH

JAC_25344_E_orarius_LazaroCard_MICH

- CIG_458_E_orarius_Ixtlahuacan_COL

CIG_460_E_orarius_Ixtlahuacan_COL

JAC_25563_E_orarius_Hwy200_MICH

JAC_25564_E_orarius_Hwy200_MICH

- CIG_341_E_orarius_Aquila_MICH

JAC_25526_E_orarius_Hwy200_MICH

0.03

Supplementary Fig. 1. Bayesian phylogenetic inference of members of the E/eutherodactylus subgenus Syrrhophus, based on the

mitochondrial loci 16S rRNA. All nodes with support of less than 0.5 are collapsed.

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Appendix 1. List of subgenus Syrrhophus specimens examined in this study. Museum codes: MZFC = Museo de Zoologia de la

Facultad de Ciencias, Universidad Autonoma de México (UNAM); UTA = Amphibian and Reptile Diversity Research Center,

University of Texas at Arlington. Field number codes: ANMO = Adrian Nieto Montes de Oca, uncatalogued at MZFC; CIG = Chris

Griinwald, uncatalogued at MZFC; JAC = Jonathan A. Campbell, uncatalogued at UTA; JHM = John Malone, uncatalogued at

UTA; JRV = Jacobo Reyes- Velasco, uncatalogued at MZFC.

Specimens Examined (7 = 495)

Eleutherodactylus albolabris (n= 20): MEXICO: Guerrero: MZFC 33025-33030 (CIG 00327-00332), 33082-33085 (CIG 00390-

00393), MZFC 33108-33109 (CIG 00441-00442), MZFC 33230 (CIG 00668), MZFC 33300-33301 (CIG 00903-00904),

MZEFC 33323 (CIG 00953), MZFC 33325-33326 (CIG 00955-00956), JAC 25586, 25642.

Eleutherodactylus. angustidigitorum (n= 20): MEXICO: Jalisco: MZFC 33127-33130 (CIG 00476-00479), MZFC 33224-33225

(CIG 00662-00663), MZFC 33386-33388 (CIG 00991-00993), JAC 24912; Michoacan: MZFC 33015-33017 (CIG 00316—

00318), MZFC 33065-33070 (CIG 00373-00378), JAC 26977.

Eleutherodactylus campi (n = 13): MEXICO: Nuevo Leon: MZFC 33195-33198 (CIG 00606-00609); UNITED STATES: Texas:

JHM 1390-1394.

Eleutherodactylus colimotl (n = 20): MEXICO: Colima: MZFC 29282 (CIG 00468), MZFC 33115-33120 (CIG 00462-00467),

MZFC 33237-33239 (CIG 00682-00684), MZFC 33299 (CIG 00901), MZFC 33329-3330 (CIG 00960-00961), JAC 30498—

30499, 30631; Michoacan: MZFC 33036 (CIG 00340), JAC 23999-24001.

Eleutherodactylus cystignathoides (n = 6): MEXICO: Veracruz: MZFC 33351-33353 (CIG 01163-01165), MZFC 33354 (CIG

01170), JAC 30000-30001.

Eleutherodactylus dennisi (n = 13): MEXICO: Tamaulipas: MZFC 33255-33261 (CIG 00822-00828), UTA 59516-59521.

Eleutherodactylus dilatus (n = 19): MEXICO: Guerrero: MZFC 33089-33094 (CIG 00405-00410), MZFC 33097 (CIG 00428),

MZEFC 33231 (CIG 00669), UTA 4017-4020, 4023-4024, 5269, 5276-5279.

Eleutherodactylus erendirae (n = 25): MEXICO: Jalisco: MZFC 33000-33008 (CIG 00300-00309), MZFC 33226-33229 (CIG

00664-00667), MZFC 33232 (CIG 00673), MZFC 33234-33235 (CIG 00679-00681); Michoacan: MZFC 29274, 33019-—

33024 (CIG 00319-00325).

Eleutherodactylus floresvillelai (n = 12): MEXICO: Michoacan: MZFC 33053-33064 (CIG 00361-00372).

Eleutherodactylus grandis (n= 1): MEXICO: Ciudad de Mexico: UTA 56845.

Eleutherodactylus grunwaldi (n = 12): MEXICO: Colima: MZFC 27467-27475, MZFC 27484, MZFC 33298 (CIG 00898): JRV

00230.

Eleutherodactylus guttilatus (n = 10): MEXICO: Guanajuato: MZFC 33367-33369 (CIG 01248-01250); San Luis Potosi: MZFC

33200-33206 (CIG 00619-00625).

Eleutherodactylus interorbitalis (n= 7): MEXICO: Sinaloa: MZFC 33186-33187 (CIG 00584-00585), MZFC 33190-33194 (CIG

00600-00604).

Eleutherodactylus jaliscoensis (n = 15): MEXICO: Jalisco: MZFC 33131-33141 (CIG 00480-00490), MZFC 33274-33276 (CIG

00861-00863), MZFC 33280 (CIG 00876).

Eleutherodactylus leprus (n= 7): MEXICO: Veracruz: MZFC 33345-33350 (CIG 01139-01144), CIG 01270.

Eleutherodactylus longipes (n= 3): MEXICO: Nuevo Leon: MZFC 33199 (CIG 00611); Querétaro: UTA 59421-59422.

Eleutherodactylus maculabialis sp. nov. (n = 27): MEXICO: Guerrero: MZFC 33307-33319 (CIG 00916-00923, 00940-00941,

00945-00947), MZFC 33321 (CIG 00949), MZFC 33323 (CIG 00953), CIG 01484-01485, 01501, JAC 25643-25646.

Eleutherodactylus marnocki (n = 3): USA: Texas: JHM 1427-1429.

Eleutherodactylus manantlanensis (n = 14): MEXICO: Colima: MZFC 33372-33377 (CIG 00530-00535), MZFC 33379-33381

(CIG 00646-00648), MZFC 33292-33296 (CIG 00892-00896).

Eleutherodactylus maurus (n = 11): MEXICO: Estado de México: MZFC 33071-33076 (CIG 00379-00384), MZFC 33355 (CIG

01174); Morelos: MZFC 33077-33080 (CIG 00385-00388).

Eleutherodactylus modestus (n = 34): MEXICO: Colima: MZFC 26888-26889, MZFC 33263-33270 (CIG 00850-00857), MZFC

33291 (CIG 00891), MZFC 33297 (CIG 00897); Jalisco: MZFC 33144-33149 (CIG 00493-00498), MZFC 33150-33154 (CIG

00505-00509), MZFC 33161 (CIG 00522), MZFC 33183-33185 (CIG 00570-00572), MZFC 33217-33223 (00655-00661).

Eleutherodactylus nebulosus (n= 9): MEXICO: Chiapas: MZFC 33249-33251 (CIG 00753, 00755-00756), MZFC 33361-33366

(CIG 01236-01241).

Eleutherodactylus nietoi (n = 13): MEXICO: Michoacan: MZFC 33121 (CIG 00299), MZFC 33042-33045 (CIG 00346-00349),

MZFC 33050-33052 (CIG 00355-00357), MZFC 33336-33337 (CIG 00974-00975), MZFC 33342-33343 (CIG 00983-—

00984), MZFC 33344 (CIG 00994).

Eleutherodactylus nitidus (n= 31): MEXICO: Estado de México: JAC 27237; Guerrero: MZFC 33096-33097 (CIG 00411-00412),

MZFC 33104-33105 (CIG 00437-00438), JAC 25815; Morelos: MZFC 33081 (CIG 00389); Oaxaca: MZFC 33357-33358

(CIG 01211-01212); Puebla: MZFC 33356 (CIG 01181), JAC 27256-27276.

Eleutherodactylus orarius (n= 13): MEXICO: Colima: MZFC 26890, MZFC 33262 (CIG 00849); Michoacan: MZFC 33037 (CIG

00341), MZFC 33335 (CIG 00973), JAC 24020, 25526, 25563-25564, 29107, 30500-30501, 30517, 30625.

Eleutherodactylus pallidus (n = 13): MEXICO: Jalisco: MZFC 33271-33272 (CIG 00858-00859); Nayarit: MZFC 33189 (CIG

00588), MZFC 33212-33216 (CIG 00650-00654), MZFC 33243-33245 (CIG 00688-00690), MZFC 33018 (CIG 00995);

Sinaloa: MZFC 33188 (CIG 00586).

Eleutherodactylus petersi (n = 25): MEXICO: Guerrero: MZFC 33034-33035 (CIG 00336-00337); JAC 25219, 25265-25266,

25299; Jalisco: MZFC 33010-33014 (CIG 00310-00314), MZFC 33034-33035 (CIG 00336-00337), MZFC 33110 (CIG

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Grunwald et al.

00457), MZFC 33273 (CIG 00860), JAC 28612; Michoacan: MZFC 33382-33385 (CIG 00675-00677), JAC 26947; Nayarit:

MZFC 33211 (CIG 00649), MZFC 33240-33242 (CIG 00685-00687).

Eleutherodactylus. pipilans (n = 15): MEXICO: Guerrero: MZFC 33086-33088 (CIG 00396—00398), MZFC 33106-33107 (CIG

00439-00440), MZFC 33322 (CIG 00952), CIG 1465; Oaxaca: MZFC 33210 (CIG 00645), JAC 24283, 25809-25811.

Eleutherodactylus rufescens (n = 40): MEXICO: Jalisco: MZFC 33122-33126 (CIG 00471-00475), MZFC 33162-33164 (CIG

00527-00529), MZFC 33165-33174 (CIG 00544-00553), MZFC 33385 (CIG 00678); Michoacan: MZFC 33038-33041 (CIG

00342-00345), MZFC 33046-33049 (CIG 00350-00353), MZFC 33175-33182 (CIG 00559-00566), MZFC 33233 (CIG

00674), MZFC 33338 (CIG 00976), MZFC 33339-33341 (CIG 00980-00982).

Eleutherodactylus saxatilis (n = 4): MEXICO: Sinaloa: MZFC 26893, 26896, 26898-26899.

Eleutherodactylus sentinelus sp. nov. (n = 8): MEXICO: Guerrero: MZFC 33031-33033 (CIG 00333-00335), MZFC 33302-

33306 (CIG 00907-00913).

Eleutherodactylus syristes (n = 21): MEXICO: Guerrero: ANMO 2999; MZFC 33098-33103 (CIG 00431-00436), MZFC 33324

(CIG 00954), MZFC 33327-33328 (CIG 00957-00958) JAC 25701-25703; Oaxaca: MZFC 33207-33208 (CIG 00627-00628),

MZFC 33209 (CIG 00644), 33378 (CIG00643), MZFC 33246-33247 (CIG 00713-00714), MZFC 33359-33360 (CIG 01232—

01233).

Eleutherodactylus teretistes (n = 5): MEXICO: Jalisco: MZFC 33142-33143 (CIG 00491-00492), MZFC 33277-33279 (CIG

00864—00866).

Eleutherodactylus verrucipes (n = 3): MEXICO: MZFC 33253-33254 (CIG 00813-00814), CIG 01273.

Eleutherodactylus wixarika (n = 3): MEXICO: Jalisco: MZFC 27477-27479.

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Two new Eleutherodactylus species from Mexico

Appendix 2. Specimen information and GenBank accession numbers.

Field number Species Museum number Locality GenBank number

— Eleutherodactylus symingtoni — Cuba: Pinar del Rio EF493643

— Eleutherodactylus nitidus AMCC118239 Mexico: Puebla EU186712

— Eleutherodactylus erythrochomus MZFC 16254 Mexico: Guerrero EU186711

— Eleutherodactylus zeus USNM335740 Cuba: Pinar del Rio EF493718

ANMO2999 Eleutherodactylus syristes — Mexico: Guerrero MT872423

CIG1142 Eleutherodactylus leprus MZFC33348 Mexico: Veracruz MT872424

CIG1144 Eleutherodactylus leprus MZFC33350 Mexico: Veracruz MT872425

CIG1232 Eleutherodactylus syristes MZFC33359 Mexico: Oaxaca MT872426

CIG1233 Eleutherodactylus syristes MZFC33360 Mexico: Oaxaca MT872427

CIG1236 Eleutherodactylus nebulosus MZFC33361 Mexico: Chiapas MT872428

CIG1237 Eleutherodactylus nebulosus MZFC33362 Mexico: Chiapas MT872429

CIG1238 Eleutherodactylus nebulosus MZFC33363 Mexico: Chiapas MT872430

CIG1240 Eleutherodactylus nebulosus MZFC33365 Mexico: Chiapas MT872431

CIG1241 Eleutherodactylus nebulosus MZFC33366 Mexico: Chiapas MT872432

CIG1270 Eleutherodactylus leprus — Mexico: Veracruz MT872433

CIG310 Eleutherodactylus petersi MZFC33010 Mexico: Jalisco MT872487

CIG311 Eleutherodactylus nitidus MZFC33011 Mexico: Jalisco MG857033

CIG314 Eleutherodactylus nitidus MZFC33014 Mexico: Jalisco MG857034

CIG330 Eleutherodactylus albolabris MZFC33028 Mexico: Guerrero MT872481

CIG331 Eleutherodactylus albolabris MZFC33029 Mexico: Guerrero MT872482

CIG332 Eleutherodactylus albolabris MZFC33030 Mexico: Guerrero MT872483

CIG333 Eleutherodactylus sentinelus sp. nov. MZFC33031 Mexico: Guerrero MT872484

CIG334 Eleutherodactylus sentinelus sp. nov. MZFC33032 Mexico: Guerrero MT872485

CIG335 Eleutherodactylus sentinelus sp. nov. MZFC33033 Mexico: Guerrero MT872486

CIG336 Eleutherodactylus nitidus MZFC33034 Mexico: Guerrero MG857032

CIG337 Eleutherodactylus petersi MZFC33035 Mexico: Guerrero MT872473

CIG341 Eleutherodactylus orarius MZFC33037 Mexico: Michoacan MG857041

CIG380 Eleutherodactylus maurus MZFC33072 Mexico: Mexico MG857011

CIG382 Eleutherodactylus maurus MZFC33074 Mexico: Mexico MT872478

CIG385 Eleutherodactylus maurus MZFC33077 Mexico: Morelos MT872479

CIG387 Eleutherodactylus maurus MZFC33079 Mexico: Morelos MT872480

CIG388 Eleutherodactylus maurus MZFC33080 Mexico: Morelos MG857010

CIG389 Eleutherodactylus nitidus MZFC3308 1 Mexico: Morelos MG857029

CIG391 Eleutherodactylus albolabris MZFC33083 Mexico: Guerrero MG856955

CIG392 Eleutherodactylus albolabris MZFC33084 Mexico: Guerrero MT872468

CIG396 Eleutherodactylus pipilans MZFC33086 Mexico: Guerrero MG857054

CIG398 Eleutherodactylus pipilans MZFC33088 Mexico: Guerrero MG857055

CIG407 Eleutherodactylus dilatus MZFC33091 Mexico: Guerrero MG856973

CIG408 Eleutherodactylus dilatus MZFC33092 Mexico: Guerrero MG856974

CIG412 Eleutherodactylus nitidus MZFC33096 Mexico: Guerrero MG857031

CIG431 Eleutherodactylus syristes MZFC33098 Mexico: Guerrero MT872471

CIG434 Eleutherodactylus syristes MZFC33101 Mexico: Guerrero MT872472

CIG435 Eleutherodactylus syristes MZFC33102 Mexico: Guerrero MG857071

CIG439 Eleutherodactylus nitidus MZFC33106 Mexico: Guerrero MT872474

CIG440 Eleutherodactylus pipilans MZFC33107 Mexico: Guerrero MT872475

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Grunwald et al.

Appendix 2 continued. Specimen information and GenBank accession numbers.

Field number Species Museum number Locality GenBank number

CIG441 Eleutherodactylus albolabris MZFC33108 Mexico: Guerrero MT872476

CIG458 Eleutherodactylus orarius MZFC33111 Mexico: Colima MT872477

CIG460 Eleutherodactylus orarius MZFC33113 Mexico: Colima MG857042

CIG544 Eleutherodactylus rufescens MZFC33165 Mexico: Jalisco MG857039

CIG563 Eleutherodactylus rufescens MZFC33179 Mexico: Michoacan MG857060

CIG611 Eleutherodactylus longipes MZFC33199 Mexico: Nuevo Leon MG857006

Mexico: San Luis

CIG619 Eleutherodactylus guttilatus MZFC33200 Potosi MG856994

CIG627 Eleutherodactylus syristes MZFC33207 Mexico: Oaxaca MT872467

C1IG628 Eleutherodactylus syristes MZFC33208 Mexico: Oaxaca MG857073

CIG649 Eleutherodactylus sp. nov. MZFC33211 Mexico: Nayarit MT872469

CIG713 Eleutherodactylus syristes MZFC33246 Mexico: Oaxaca MT872470

CIG714 Eleutherodactylus syristes MZFC33247 Mexico: Oaxaca MG857072

CIG715 Eleutherodactylus nitidus MZFC33248 Mexico: Oaxaca MG857030

CIG753 Eleutherodactylus nebulosus MZFC33249 Mexico: Chiapas MG857056

CIG755 Eleutherodactylus nebulosus MZFC33250 Mexico: Chiapas MG857057

CIG813 Eleutherodactylus verrucipes MZFC33253 Mexico: Tamaulipas MG857079

CIG857 Eleutherodactylus modestus MZFC33270 Mexico: Colima MG857021

CIG893 Eleutherodactylus manantlanensis MZFC33293 Mexico: Colima MG857007

Eleutherodactylus maculabialis

CIG921 sp. nov. MZFC33312 Mexico: Guerrero MT872460

Eleutherodactylus maculabialis

CIG923 sp. nov. MZFC33314 Mexico: Guerrero MT872461

Eleutherodactylus maculabialis

CIG940 sp. nov. MZFC33315 Mexico: Guerrero MT872462

Eleutherodactylus maculabialis

CIG941 Sp. nov. MZFC33316 Mexico: Guerrero MT872463

Eleutherodactylus maculabialis

CIG946 Sp. nov. MZFC33318 Mexico: Guerrero MT872464

Eleutherodactylus maculabialis

CIG947 Sp. nov. MZFC33319 Mexico: Guerrero MT872465

Eleutherodactylus maculabialis

CIG949 Sp. nov. MZFC33321 Mexico: Guerrero MT872466

CIG953 Eleutherodactylus albolabris MZFC33323 Mexico: Guerrero MG856956

CIG954 Eleutherodactylus syristes MZFC33324 Mexico: Guerrero MG857070

CIG984 Eleutherodactylus nietoi MZFC33343 Mexico: Michoacan MG857028

JAC24020 Eleutherodactylus orarius UTAAS9508 Mexico: Michoacan MT872434

JAC24283 Eleutherodactylus pipilans UTAA64104 Mexico: Oaxaca MT872435

JAC25219 Eleutherodactylus petersi UTAA61561 Mexico: Guerrero MT872436

JAC25265 Eleutherodactylus petersi UTAA61562 Mexico: Guerrero MT872437

JAC25266 Eleutherodactylus petersi UTAA61563 Mexico: Guerrero MT872438

JAC25299 Eleutherodactylus petersi UTAA61568 Mexico: Guerrero MT872439

JAC25341 Eleutherodactylus orarius UTAA62400 Mexico: Michoacan MT872440

JAC25342 Eleutherodactylus orarius UTAA62401 Mexico: Michoacan MT872441

JAC25343 Eleutherodactylus orarius UTAA62402 Mexico: Michoacan MT872442

JAC25344 Eleutherodactylus orarius UTAA62403 Mexico: Michoacan MT872443

JAC25526 Eleutherodactylus orarius UTAA62090 Mexico: Michoacan MT872444

JAC25563 Eleutherodactylus orarius UTAA61575 Mexico: Michoacan MT872445

JAC25564 Eleutherodactylus orarius UTAA61576 Mexico: Michoacan MT872446

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Two new Eleutherodactylus species from Mexico

Appendix 2 continued. Specimen information and GenBank accession numbers.

Field number Species Museum number Locality GenBank number

JAC25586 Eleutherodactylus albolabris UTAA61577 Mexico: Guerrero MT872447

JAC25642 Eleutherodactylus albolabris UTAA61578 Mexico: Guerrero MT872448

Eleutherodactylus maculabialis

JAC25643 Sp. nov. UTAA62091 Mexico: Guerrero MT872449

Eleutherodactylus maculabialis

JAC25644 Sp. nov. UTAA64131 Mexico: Guerrero MT872450

Eleutherodactylus maculabialis

JAC25645 Sp. nov. UTAA64130 Mexico: Guerrero MT872451

Eleutherodactylus maculabialis

JAC25646 sp. nov. — Mexico: Guerrero MT872452

JAC25701 Eleutherodactylus syristes UTAA61580 Mexico: Guerrero MT872453

JAC25702 Eleutherodactylus syristes UTAA61581 Mexico: Guerrero MT872454

JAC25703 Eleutherodactylus syristes UTAA61582 Mexico: Guerrero MT872455

JAC25809 Eleutherodactylus pipilans UTAA62404 Mexico: Oaxaca MT872456

JAC25810 Eleutherodactylus pipilans UTAA62405 Mexico: Oaxaca MT872457

JAC25811 Eleutherodactylus pipilans UTAA62406 Mexico: Oaxaca MT872458

JAC25815 Eleutherodactylus nitidus UTAA61584 Mexico: Guerrero MT872459

JRV142 Eleutherodactylus grunwaldi — Mexico: Jalisco MG856993

JRV160 Eleutherodactylus wixarika MZFZ27477 Mexico: Jalisco MG857081

Amphib. Reptile Conserv. 34 February 2021 | Volume 15 | Number 1 | e272

Grunwald et al.

Appendix 3. Proposed standardized common names for the E/eutherodactylus nitidus species group members.

Eleutherodactylus albolabris White-lipped Whistling Frog

Eleutherodactylus dilatus Omiltemi Peeping Frog

Eleutherodactylus erythrochomus Tierra Colorada Peeping Frog

Eleutherodactylus maurus Dusky Piping Frog

Eleutherodactylus nitidus Shiny Whistling Frog

Eleutherodactylus orarius Coastal Whistling Frog

Eleutherodactylus petersi Peters’ Whistling Frog

Eleutherodactylus pipilans Guerrero Peeping Frog

Eleutherodactylus nebulosus Clouded Peeping Frog

Eleutherodactylus maculabialis sp. nov. Spot-lipped Trilling Frog

Eleutherodactylus sentinelus sp. nov. El Balsamo Peeping Frog

Eleutherodactylus syristes Oaxaca Trilling Frog

Amphib. Reptile Conserv. 35 February 2021 | Volume 15 | Number 1 | e272

a gill EB CO, ~

Official journal website:

amphibian-reptile-conservation.org

Amphibian & Reptile Conservation

15(1) [General Section]: 36—56 (e273).

The distribution and conservation status of the

Dwarf Marsupial Frog (Flectonotus fitzgeraldi, Anura,

Hemiphractidae) in Trinidad, Tobago, and Venezuela

‘Joanna Smith, *Michael J. Jowers, *Renoir J. Auguste, “Paul Hoskisson, "Cammy Beyfts, *Greig M.

Muir, °Mark S. Greener, ‘Dan Thornham, ‘Isabel Byrne, ’Rick Lehtinen, “Meredith Eyre, *Michael G.

Rutherford, °John C. Murphy, ‘“Mayke De Freitas, “Gilson A. Rivas, and °J. Roger Downie

'School of Natural Sciences, Bangor University, Bangor Wales LL57 2DG, UNITED KINGDOM ?CIBIO/InBIO Research Center in Biodiversity

and Genetic Resources, Porto University, Vairdo, PORTUGAL *Department of Life Science, University of the West Indies, St. Augustine, TRINIDAD

4Strathclyde Institute of Pharmacy and Biomedical Sciences, University of Strathclyde, Glasgow, Scotland G4 ORE, UNITED KINGDOM “Institute

of Evolutionary Biology, School of Biological Sciences, University of Edinburgh, Edinburgh, Scotland, UNITED KINGDOM °School of Life

Science, Graham Kerr Building, University of Glasgow, Glasgow, Scotland G12 SQ0Q, UNITED KINGDOM ‘Department of Biology, The College

of Wooster, Wooster Ohio 44691, USA *26 George Street, Innerleithen, Scotland EH44 6LH, UNITED KINGDOM °Science and Education, Field

Museum of Natural History, 1400 Lake Shore Drive, Chicago, Illinois 60616, USA '°13 Levitt Lane, Waterbeach, Cambridge, CB25 9AZ, UNITED

KINGDOM ''Museo de Biologia, Facultad Experimental de Ciencias, Universidad del Zulia, Apartado Postal 526, Maracaibo 4011, VENEZUELA

Abstract.— The Dwarf Marsupial Frog, Flectonotus fitzgeraldi (family Hemiphractidae), has been reported to occur

only in Trinidad, Tobago, and the Paria Range of Venezuela. This species is listed as Endangered on the IUCN Red

List, based on its small geographic distribution and the fragmentation of its habitat, which is said to be declining

both in extent and quality. Using molecular methods, we confirm herein that the three populations do belong to

the same species. However, extensive presence/absence and focused population surveys show that the frog’s

distribution is more extensive than previously reported in both Trinidad and Venezuela. In Trinidad and Tobago,

the frog is abundant in forests wherever its host plants occur, notably the bromeliad Heliconia bihai (Balisier in

Trinidad; Bijao in Venezuela) and the aroid Xanthosoma jacquinii (Elephant’s Ear). In Venezuela, the species is

frequently found where there is suitable habitat, but an exhaustive population study is needed to diagnose its

current situation. No evidence was found of habitat decline in Trinidad and Tobago, but in Venezuela the loss of

habitat is evident, mainly because of subsistence agricultural activities, which have been developing in northeast

Venezuela since at least 1930. The Red List status of this species is in need of revision.

Keywords. Amphibia, habitat loss, IUCN Red List, phytotelmata, southern Caribbean

Resumen.— La ranita marsupial Flectonotus fitzgeraldi (familia Hemiphractidae), se conoce solo de Trinidad,

Tobago y la Region de Paria en Venezuela. La Lista Roja de la UICN cataloga esta especie como En Peligro,

con base a su area de distribucion geografica reducida y a la fragmentacion de su habitat, el cual se reduce

progresivamente tanto en su extension como en su calidad. Usando métodos moleculares comparativos, en

este trabajo se confirma que las tres poblaciones pertenecen a una misma especie. Sin embargo, un estudio

poblacional y de presencia/ausencia demuestra que la distribuciOn geografica de este anfibio es mas amplia

que la senalada anteriormente, principalmente en Trinidad y en Venezuela. En Trinidad y Tobago, esta rana es

frecuente en areas boscosas siempre que sus plantas huespedes esten presentes, principalmente bromelias,

Heliconia bihai (balisier en Trinidad; bijao en Venezuela) y Xanthosoma jacquinii (oreja de elefante). En Venezuela,

la especie es frecuente, pero se requiere un estudio poblacional mas exhaustivo para diagnosticar con precision

su situacion. No se encontro evidencia de un deterioro importante en la calidad de su habitat en Trinidad y

Tobago, pero en Venezuela la péerdida de habitat es evidente, principalmente como resultado de las actividades

agricolas de subsistencia, que se desarrollan en el noroeste de Venezuela al menos desde 1930. El estado de esta

especie en la Lista Roja precisa de una revision.

Palabras clave. Anfibia, perdida de habitat, Lista roja IUCN, fitotelmata, Sur del Caribe

Citation: Smith J, Jowers MJ, Auguste RJ, Hoskisson P, Beyts C, Muir GM, Greener MS, Thornham D, Byrne I, Lehtinen R, Eyre M, Rutherford MG,

Murphy JC, De Freitas M, Rivas GA, Downie JR. 2021. The distribution and conservation status of the Dwarf Marsupial Frog (Flectonotus fitzgeraldi,

Anura, Hemiphractidae) in Trinidad, Tobago, and Venezuela. Amphibian & Reptile Conservation 15(1) [General Section]: 36-56 (e273).

Correspondence. *michaeljowers@hotmail.com (MJJ), j.smith@bangor.ac.uk (JS), Renguste@gmail.com (RJA), paul. hoskisson@strath.

ac.uk (PH), s1437006@sms.ed.ac.uk (CB), gmuir66@talktalk.net (GM), mark.s.greener@gmail.com (MSG), d.thornham@bangor.ac.uk (DT),

isabelbyrne 11@gmail.com (1B), NONE (ME), RLEHTINEN@wooster.edu (RL), Mike. Rutherford@sta.uwi.edu (MGR), serpentresearch@gmail.

com (JCM), mayvkef@gmail.com (MDF), anolis30@hotmail.com (GAR), Roger. Downie@glasgow.ac.uk (JRD)

Amphib. Reptile Conserv. 36 June 2021 | Volume 15 | Number 1 | e237

Smith et al.

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: 29 September 2020; Published: 30 June 2021

Introduction

The neotropical frog family Hemiphractidae includes

six tree frog genera (118 species described so far; Frost

2020), in which females incubate small clutches of

large eggs on their backs, either openly (in Stefania and

Cryptobatrachus) or in a more or less closed cutaneous

pouch (all other genera). One member of a _ two-

species genus, the Dwarf Marsupial Frog Flectonotus

fitzgeraldi (Parker 1934), is found in Trinidad, Tobago,

and the nearby Paria Biota region of Venezuela, which

includes the Serrania de Paria, Cerros Campeare, and

La Cerbatana. In this small species (maximum snout-

vent length in males 21 mm and in females 25 mm),

developing individuals (clutch size 2-6) hatch as

late-stage larvae and are deposited by the female into

phytotelmata, which are small bodies of water enclosed

by leaves, such as bromeliad ‘tanks.’ These larvae

do not feed but subsist on the remaining yolk (Kenny

1969) and soon enter the process of metamorphosis,

which is indicated by forelimb emergence. Downie et

al. (2004) found that metamorphosis was complete (tail

reabsorption) after 4.5 + 0.7 days. Duellman and Gray

(1983) suggested that individual Flectonotus females

were capable of producing several egg clutches in

a year, but data are still needed to test this suggestion

for F. fitzgeraldi. Flectonotus fitzgeraldi are arboreal at

night, with males calling from leaves such as those of

bromeliads and Heliconia bihai, which 1s said to occur

primarily during a period of about one hour, from soon

before until soon after sunset (Murphy 1997). During

the day, they may be active in leaf litter on the ground

(Duellman and Gray 1983).

Flectonotus fitzgeraldi was assessed as Endangered

by La Marca et al. (2004) for the IUCN Red List, based

on criteria Blab(iil), on the grounds that its “extent of

occurrence is less than 5,000 km7*, its distribution is

severely fragmented, and that there 1s continuing decline

in the extent and quality of its habitat” in all three

regions. Its population status was judged to be declining.

However, this assessment appears not to have been

based on published research and included the statement

that “further work is needed to establish the current

population status of this species.”

In the past, many anuran species were listed as

occurring in Trinidad, Tobago, and northern Venezuela.

However, modern assessments using molecular methods

have not always supported these patterns. For example,

the aromobatid Mannophryne trinitatis is now known

to occur only in Trinidad, with the Venezuelan and

Tobagonian populations assigned to M. venezuelensis and

M. olmonae, respectively (Barrio-Amoros et al. 2006;

Amphib. Reptile Conserv.

Manzanilla et al. 2007, 2009). It is important, therefore,

especially for a species classified as Endangered, to be

certain that the same species occurs in all three areas.

Kenny (1969) reported the distribution of F: fitzgeraldi

(then known as a member of the genus Nototheca) from

surveys made during 1955-1961. He listed the frog

as occurring “throughout Trinidad from sea level to

the highest peaks,” and listed several locations in the

Northern Range (including the summit of El Tucuche,

Trinidad’s second highest mountain, the Aripo Valley,

the mouth of Grande Riviere river, and Matelot); other

locations were Rio Claro, Mayaro, and Tamana Hill.

Later, Clarke et al. (1995) confirmed the presence of F°

fitzgeraldi in two species of bromeliads on the summit

of El Tucuche, but did not find it on Cerro del Aripo.

Murphy (1997) added the Arima Valley, the road to

Arena Dam, and Morne Bleu Ridge to the list of Trinidad

records. He also listed Tobago locations at Northside

Road, Hillsborough Dam, Speyside, and the Windward

Road by Lambeau Crown Trace. The distribution map

accompanying the IUCN Red List assessment (La Marca

et al. 2004) shows F: fitzgeraldi occupying an area of

4.500 km? in all. This included areas in northeastern

Venezuela, adjacent to Trinidad, of approximately 3,700

km/?; an area of Trinidad, mainly in the central part of the

Northern Range but extending a little south, of about 600

km’; and most of the northeast of Tobago, centered on

the Main Ridge forest, of about 200 km’. A Field Guide

to the Amphibians and Reptiles of Trinidad and Tobago

(Murphy et al. 2018) gives a similar picture for Tobago,

but extends the Trinidad distribution somewhat to the

west, east, and south, based on museum specimens and

published records.

The aims of this paper are three-fold: a) to test whether

the three populations of F fitzgeraldi can safely be

assigned to a single species using molecular methods; b)

to collate recent data on the distribution and population

size of F fitzgeraldi, some published and some new in

this paper, and c) to assess whether this species indeed

qualifies for its current Endangered (EN) status.

Materials and Methods

Flectonotus fitzgeraldi specimens were collected at

locations in Trinidad and Tobago, and Venezuela, under

licenses from the Trinidad and Tobago Government

Wildlife Section: Special Game Licenses issued for

scientific purposes in 2012-2019 to John Murphy,

Mike Rutherford, and Rick Lehtinen; and in Venezuela

under the collection permit numbers 1375 and 0878

granted to Gilson A. Rivas by the Republica Bolivariana

de Venezuela’s Ministerio del Poder Popular para

June 2021 | Volume 15 | Number 1 | e237

Flectonotus fitzgeraldi in Trinidad, Tobago, and Venezuela

Ecosocialismo y Aguas. Voucher specimens are

deposited in the Museo de Biologia, Universidad del

Zulia, Maracaibo (MBLUZ); Museo de la Estacion

Biologica de Rancho Grande, El Limon (EBRG), Museo

de Historia Natural La Salle, Caracas (MHNLS), Rick

Lehtinen field number (RML), and Museum of Zoology,

University of West Indies (UWIZM). Animals used for

genetic analysis were euthanized following the ASIH

guidelines using MS-222 in water.

DNA Analysis

Adults and tadpoles of Filectonotus fitzgeraldi were

caught with hand nets. The DNA was sequenced from

11 tadpole tail tips and adults (Table 1). Whole genomic

DNA was extracted using the DNeasy Blood and

Tissue kit (QIAGEN, Hilden, Germany) following the

manufacturer’s instructions. One mitochondrial gene

fragment, 16S rDNA, was used with primers 16SL and

16SH (Palumbi 1991) and the 16S primers reported

by Wiens et al. (2005). Templates were sequenced on

both strands, and the complementary reads were used

to resolve rare, ambiguous base-calls in Sequencher

v4.9 (Gene Codes Corporation, Ann Arbor, Michigan,

USA). The lengths of the sequences were not the

same for all individuals as different primers were used

in different laboratories. All Flectonotus 16S rDNA

sequences were downloaded from GenBank (n = 5), but

the final alignment included only four of them as the

only Brazilian specimen (Flectonotus sp.) showed very

high divergence from all other species. The total dataset

consisted of 15 individuals, including two individuals

of Flectonotus pygmaeus from Venezuela which were

used as an outgroup. Sequences were aligned in Seaview

v.4.2.11 (Gouy et al. 2010) under ClustalW2 (Larkin

et al. 2007) with default settings. The final alignment

comprised 859 base pairs.

The most appropriate substitution model (TrN) was

determined by the Bayesian Information Criterion (BIC)

in jModeltest 2 (Posada 2008). Phylogenetic analyses

were run to assess the relationships between localities,

and taxa were inferred using the Bayesian Inference (BI)

optimality criterion under the best-fitting substitution

model. MrBayes v3 (Ronquist and Huelsenbeck 2003)

was used with default priors and Markov chain settings,

and with random starting trees. Each run consisted of

four chains of 10 million generations, sampled every

10,000 generations. Phylogenetic relationships were also

estimated using a Maximum Likelihood (ML) approach,

as implemented in the software RAxML v7.0.4 (Silvestro

and Michalak 2012), under the GTR model. All analyses

were performed using the CIPRES platform (Miller et

al. 2010). A median-joining haplotype network (Bandelt

et al. 1999) was constructed using Popart v.1.7 (Leigh

and Bryant 2015). Because of missing data in the original

alignment, a shorter alignment (500 bp) was used for the

network analysis.

Amphib. Reptile Conserv.

Survey Methods

The temporal pattern of calling. To establish the

temporal pattern of calling in F. fitzgeraldi, in June 2006

an undisturbed site in Trinidad’s Northern Range was

chosen where the species had previously been recorded

(Morne Bleu Ridge, Arima Valley, 10°43’37”N;

61°18’21”W, elevation 730 m; Murphy 1997). Starting

from the radio-transmitter station at the west end of

the trail, 20 sequential 100 m transects were marked

out eastwards, using colored tape tied to trees. Trained

observers walked the trail in pairs a total of eight times

from June to August, from late afternoon until 2000 h,

listening for both F fitzgeraldi and Pristimantis urichi.

Each 100-m transect required about 9 min to survey.

Frog calls judged to be made from within a distance

of approximately 5 m on either side of the trail were

recorded (the numbers of frogs per transect were used

for population estimates, as noted below, and this survey

was repeated in June — August 2007). Since these species

both call as dispersed individuals rather than in multi-

individual choruses, a slow walk is a reliable way to

estimate the number of calling frogs. To ensure there was

no temporal bias in recording along the 2-km trail, the

start times of the 20 transects were randomized.

Presence/absence surveys. Trinidad and Tobago: The

observers who contributed to this study learned the call

of F: fitzgeraldi either by listening to recordings made by

Morley Read (no date recorded) or by Paul Hoskisson,

or by visiting a site where F: fitzgeraldi is common and

listening to the call at dusk, while also scanning the

vegetation to obtain a sighting of the frogs. The surveys

reported here used the frog’s call as an evidence of

presence, since searching for the frogs more directly can

cause disturbance and may interfere with calling. Frogs

were sometimes seen when listening was accompanied

by torch use, but sightings were not used as a main

criterion for presence. Surveys generally began just as

light was fading, at about 1820 h, and continued until

about 1930 h, when the calling declined in frequency.

Some surveys were made by road from a vehicle,

stopping to listen for calls. In this case, stops were made

whenever a stand of Heliconia bihai was seen close to

the road edge, since this plant had a strong association

with F: fitzgeraldi. However, in the absence of H. bihai,

stops were made every few hundred meters, since the

frog also occurs in bromeliads on trees. Other surveys

reported here were a component of regional Bioblitzes

(DITOS consortium 2017) organized by the Trinidad

and Tobago Field Naturalists’ Club and the University of

the West Indies Zoology Museum (2012-2019), which

aimed to record every species encountered in an area

over a 24-hour period. Two of the authors (John Murphy

and Renoir Auguste) coordinated the herpetology group

during these surveys, which involved a small team of

observers walking slowly along forest trails from dusk

June 2021 | Volume 15 | Number 1 | e237

Smith et al.

Table 1. Flectonotus fitzgeraldi vouchers, localities, and GenBank accession numbers.

Voucher Locality GenBank accession number

MBLUZ 0395 Venezuela, Cerro Humo, Peninsula de Paria MT968884

MBLUZ 0448 Venezuela, Cerro Humo, Peninsula de Paria MT968885

EBRG 7337 Venezuela, Cerro La Cerbatana MT968886

EBGR 7346 Venezuela, Cerro Campeare MT968887

UWIZM.2015.18.4 Trinidad, Aripo Savanna MT968892

UWIZM.2019.36 Trinidad, Tamana MT968883

RMLO115 Tobago, Forest Reserve MT968888

RMLO116 Tobago, Forest Reserve MT968889

RMLO117 Tobago, Forest Reserve MT968890

RMLO118 Tobago, Forest Reserve MT968891

UWIZM.2012.27.23 Tobago, Charlotteville (Flagstaff hill) MT968882

onwards, listening for any frog calls, including those of

EF fitzgeraldi. Another approach was the use of transects,

where the number of frogs heard calling was recorded

along a fixed length of trail. In some cases, audio

recordings were made. Locations were recorded using a

hand-held GPS device.

Venezuela: Data were assembled from the published

literature and a revision of Venezuelan museum

specimens (see specimen collection information above

and Specimens Examined, Appendix 1).

Maps recording presence/absence were created with

QGIS_ v3.0.2 software (https://www.qgis.org/en/site/)

in WGS84 datum, using as a base layer the Google

(obtained through the Web Mapping Service in QGIS).

GPS coordinates were contributed by the authors (see

Supplementary Fig. Sl). The extents of occurrence

for F. fitzgeraldi were calculated for Trinidad, Tobago,

and Venezuela individually using QGIS (version 3.0.2,

Girona). GPS coordinates where the species was present

were used to create a minimum convex polygon around

the outermost coordinates within the country using the

QGIS minimum bounding geometry tool applied using

convex hull as the selected geometry type. The convex

polygons thus created were clipped to their respective

country’s shape file so that areas of the polygons which

extended into the sea were excluded. The areas of the

clipped polygons were subsequently calculated to provide

the extent of occurrence of the species for Trinidad,

Tobago, and Venezuela in km? (see Supplementary Fig.

S1).

Population estimates. Population densities were

estimated for Flectonotus fitzgeraldi along Trinidad’s

Morne Bleu Ridge in 2006 and 2007 using transects at

dusk, as described above (temporal pattern of calling).

The number of calling males along a 1 00-m transect which

was 10 m wide (1,000 m*) was extrapolated to give the

Amphib. Reptile Conserv.

density in frogs per hectare. The site was surveyed again

in 2012, 2013, and 2014, using the same methodology,

except that only the first 150 m of the trail were surveyed.

No specific population estimates were made in Tobago or

in Venezuela (see Table 2 and comments in Discussion).

Results

DNA analysis. All specimens from Venezuela (Paria

Biota region) recovered the same haplotype, while the

Trinidad and Tobago frogs recovered two and three

haplotypes, respectively (Fig. 1). The phylogenetic

analyses recovered a _ well-supported monophyletic

clade of all Flectonotus fitzgeraldi with low genetic

differentiation throughout the nominal species range,

reflecting a low genetic variability throughout. Only the

Venezuelan frogs were monophyletic.

The temporal pattern of calling. Figure 2 shows the

temporal pattern of calling at Morne Bleu Ridge, Trinidad,

for both Flectonotus fitzgeraldi and Pristimantis urichi.

The figure shows clearly that while single F’ fitzgeraldi

individuals may occasionally call in the afternoon,

almost all call activity 1s concentrated within a narrow

time period between 1800 h and 1915 h. The pattern of

P. urichi serves as a comparison to the dusk-centered

calling pattern of F’ fitzgeraldi, and to show that the short

calling period of F: fitzgeraldi is not due to some external

factor such as weather.

Presence/absence surveys. Presence/absence survey

results are shown in Table 2 and Fig. 3. GPS locations

are given in Supplementary Table S1. For Trinidad and

Tobago, most of the results are derived from survey data

published here for the first time, with a few data points

drawing on recently published papers. In Trinidad, we

only failed to find any Flectonotus fitzgeraldi in a few

surveyed locations (mainly Icacos in the southwest, and

June 2021 | Volume 15 | Number 1 | e237

Flectonotus fitzgeraldi in Trinidad, Tobago, and Venezuela

@ Tobago 1

@ Trinidad

(O11 5)

IZM.2012.27.23)

(0.98 iam

0118)

[7M .2019.36)

M.2015.18.4)

100/1

Flectonotus fitzgeraldi

uela (MBLUZ 0448)

zuela (MBLUZ 0395)

88/-

ruela (EBRG 7337)

ruela (EBRG 7346)

1400/1 F. pygmaeus Venezuela (DO679382)

—— FF. oygmaeus Venezuela (KR270418)

Fig. 1. Best Maximum Likelihood (ML) tree of Flectonotus fitzgeraldi populations from Trinidad, Tobago, and Venezuela. Values at

the nodes are comprised of ML posterior probabilities (> 75%) and Bayesian Inference probabilities (> 95%). In the Medium-Joining

network, numbers in circles represent the number of sequences with the same haplotype and dashes are number of substitutions.

the Port of Spain conurbation in the northwest). The

largest areas remaining to be surveyed are in the south-

west and south-central regions. In Tobago, the northeast

has been surveyed extensively and F: fitzgeraldi has

been found throughout. However, central Tobago is

mountainous with few roads, and a large area remains

unsurveyed. In Venezuela, systematic surveys were not

conducted. Nevertheless, F’ fitzgeraldi is reported as

occurring throughout the Paria Peninsula and further

west in northern Venezuela, including Cerros Campeare

and La Cerbatana as well as the Turimiquire massif,

as predicted by Rivas et al. (2018), and including sites

from near sea level to 1,200 m asl. Most records from

Venezuela come from disturbed locations within natural

habitat areas, mainly along the edges of patches cleared

for shifting agriculture or in gardens near paved roads.

Our observations in the northeastern portion of the

country were made at night; most frogs were detected

visually, except six specimens that were heard calling

at night (ca. 2000 h) in Campeare on 13 November

2016. Six specimens were observed staying on leaves

of Dracaena, an ornamental plant (Asparagaceae),

Amphib. Reptile Conserv.

during a survey that was carried out for visual inspection

between 1700 and 2100 h. Specimens from La Cerbatana

were observed resting inside the axil of a bromeliad,

presumably Tillandsia sp. At Cerro Humo, individuals

were observed on large leaves, possibly belonging to the

aracean genus Xanthosoma.

Although these surveys were not intended to estimate

population sizes, the number of calling males at any

particular site was recorded in both Trinidad and Tobago

in 2019. While the population of F fitzgeraldi present

was not quantified, the numbers of calling males were

often considerable, especially in extensive stands of

Heliconia bihai, with up to 35 being counted at Flagstaff

Road in Tobago.

Population estimates. Population estimates for Morne

Bleu Ridge, Trinidad, are shown in Table 3. The

estimates are broadly similar across the years, except for

2006, which was the first year of these surveys, when

the methodology was still being developed. The team

leader for that year (JS) considers that the low average

numbers resulted from the surveys starting too early in

June 2021 | Volume 15 | Number 1 | e237

Smith et al.

mD ©

Maximum number of frogs calling/10min

14:00 15:00 16:00 17:00 18:00 19:00 20:00

Time of day

Fig. 2. Maximum numbers of frogs calling per 10 minutes during eight transects along the Morne Bleu ridge in 2006 (filled circles:

Flectonotus fitzgeraldi; empty circles: Pristimantis urichi).

Fig. 3A. Map showing both presence and absence of Flectonotus fitzgeraldi as reported in this study in Trinidad. Location numbers

correspond to those in Table 2.

Amphib. Reptile Conserv. 41 June 2021 | Volume 15 | Number 1 | e237

Flectonotus fitzgeraldi in Trinidad, Tobago, and Venezuela

Fig. 3B. Map showing both presence and absence of Flectonotus fitzgeraldi as reported in this study in Tobago. Location numbers

correspond to those in Table 2.

F. fitzgeraldi

@ Present

Fig. 3C. Map showing both presence and absence of Flectonotus fitzgeraldi as reported in this study in Venezuela. Location

numbers correspond to those in Table 2.

Amphib. Reptile Conserv. 42 June 2021 | Volume 15 | Number 1 | e237

Smith et al.

Table 2. Results from presence/absence surveys for F/ectonotus fitzgeraldi in A) Trinidad; B) Tobago, 2006—2019; and C) Venezuela.

Numbers in parentheses following locations refer to numbers on the maps (Fig. 3). GPS co-ordinates for all locations are given in

Supplementary Material 1. Superscripts in the ‘‘ Year” column indicate sources of data: 'Ogilvy et al. 2007; *Smith 2008; *Greener

2015; “Bioblitz species lists (Rutherford 2013, 2014, 2017, 2018 a,b,c); *Authors’ unpublished field notes; “Auguste et al. 2015,

Auguste and Hailey 2018; 7Auguste 2019; "Mohammed et al. 2014; ?Eyre 2013; '°Rivas’ unpublished field notes; ''Rivas et al. 2018;

'Barrio-Amoros et al. 2019; '*Duellman and Gray 1983; "MBLUZ; SEBRG; '“MHNLS.

A) Trinidad

Location

Arima Valley (1A): Morne

Bleu

(1B): Simla

(1C): Springhill Estate

Aripo Savanna (2)

Brasso Seco (3)

Caroni Swamp, northern

section (4)

Chatham (5)

Cumuto-St. Raphael Road (6)

Caura Valley (7)

Edith Falls (8)

Grand Riviere (9)

Guaico-Tamana Road (10)

Icacos (11)

Lopinot Valley (12)

Matura Beach Road (13)

Matura Forest (14)

Moruga (15)

Mount St. Benedict (16)

Nariva Swamp (17)

Port of Spain (18)

Rio Claro, roads south (19)

Tabaquite (20)

Tableland (21)

Tamana Cave (22)

Toco (23)

Trinity Hills (24)

Tucker valley (25)

Amphib. Reptile Conserv.

Year

2006!, 20077,

20127: 201327,

20143, 20195

As above

2006!, 20077

201'5°

2019°

20164

2019"

2019°

20123, 20133,

20143, 2019°

2019°

20174, 20195

2019°

20167, 2019°

2019°

20123, 20137,

20148

2014*, 2016’

20164

2019°

2019*

2019°

20184

2014%, 20167

20124

Survey type

Audio path transects;

Bioblitz

Audio and visual path

transects

Audio path transects

Bioblitz

Audio transects

Audio road transect

Audio transects

Audio path transects

Audio road transect

Bioblitz; audio road

transects

Audio road transects

Audio road transect

Audio path transects

Audio transects

Bioblitz; audio and _ visual

transects

Bioblitz

Audio road transect

Bioblitz

Audio transect

Audio and visual path

transect

Bioblitz

Audio road transects

Bioblitz

Findings on F- fitzgeraldi

Always present at Morne Bleu ridge,

Springhill Estate, Simla

Abundant in marsh forest, but absent from

savanna. Voucher collected 2015

Present

Absent

Present wherever Heliconia bihai occurred

(common) and in large bromeliads on trees,

Arena Forest entrance

Present at two roadside sites, both before

village; seven other locations

Present; four locations

Present

Present at several sites between Cunaripo

and Nestor, and along track to Tamana Hill,

wherever H. bihai present

Absent: forest trails by Grand lagoon;

Southern Main Road

Present; three locations

Absent

Present

Heard at Bush Bush, but not at Kernahan or

Plum Mitan

Absent at all sites checked (Botanics, St

Ann’s, Mount Hololo, Lady Chancellor)

Present; 12 locations

Present

Present; three locations. Voucher collected

2019

Present: Cumana forest trail

Heard at Edward Trace, 2014 but not in

2016

Absent

June 2021 | Volume 15 | Number 1 | e237

Flectonotus fitzgeraldi in Trinidad, Tobago, and Venezuela

B) Tobago

Location

Bloody Bay stream (1)

Cambleton (2)

Charlotteville (3)

Doctor’s river (4)

Flagstaff hill (5)

Hermitage river (6)

Louis D’Or streams (7)

Main Ridge, Spring trail (8)

Main Ridge, Gilpin trail (9)

Main Ridge, central (10)

C) Venezuela

Location

Cachipal (1)

Camino desde Macuro-a Los

Chorros (2)

Cerro Campeare (3)

Cerro La Cerbatana (4)

La Margarita (5)

Macuro (6)

Mauraco (7)

Marauquito (8)

Quebrada Las Melenas (9)

Uquire (10)

Cerro Humo (11)

Year

201272019?

2019°

2015*, 2019°

2019°

2012°, 2019°

2019°

201.9%

2019°

20109, 2012?

Year

2017104

2003'°

9016101115

9016'15

2013

2013/2

1971315

1978'°

2002, 20031116

1993)

901 510.14

Survey type

Daytime plant searches;

audio path transects

Audio and visual transects

Bioblitz; audio transects

Audio and visual transects

Audio road transects

Audio and visual transects

Audio path transects

Daytime plant searches;

audio path transects

Survey type

Visual transects

Specimen collection

Visual transect

Specimen collection

Specimen observed

Specimen collection

Visual transects

Specimen collection

Visual transect

Findings on F- fitzgeraldi

Present in XYanthosoma; heard from H. bihai

away from stream

Abundant in H. bihai all along trail; absent

from bamboo

None heard or seen along various forest

trails, 2015, 2019.

Seen and heard among H. bihai, three visits

Abundant in roadside H. bihai stands; absent

from bamboo. Voucher collected 2012

Abundant in H. bihai, not overhanging the

stream, three visits

Present among H. bihai mainly, two visits

Absent; few H. bihai in this dark closed

canopy site; two visits

Heard among streamside H. bihai

Present in Xanthosoma, heard among

streamside H. bihai. Vouchers collected

2010

Findings on F. fitzgeraldi

Present; vouchers collected

Present: vouchers collected

Several seen calling from Dracaena plants

Present: vouchers collected

Present; vouchers collected

Present: photographed

Present: vouchers collected

Present

Present: vouchers collected

Table 3. Number of calling males of Flectonotus fitzgeraldi recorded at Morne Bleu Ridge, Trinidad. Data represent means based

on eight surveys in 2006 and 2007; and five surveys in 2012-2014.

Year

2006

2007

2012

2013

2014

Amphib. Reptile Conserv.

Mean (+ SE)

19.5.+4,7

44+ 35.1

Sie15

49 +29

64 +27

Maximum

105

June 2021 | Volume 15 | Number 1 | e237

Smith et al.

the evenings, before the calling pattern had been fully

determined. Mean as well as maximum numbers are

shown. These numbers are for calling males only, so

they must be considerable underestimates of the actual

populations. Taken together, the mean and maximum

numbers suggest that no significant change has occurred

over the nine-year span of these surveys.

Extent of occurrence. Maps showing the extent of

occurrence in each country are shown in Supplementary

Fig. Sl. The areas of occurrence are Trinidad: 2,922.4

km*; Tobago: 30.1 km?; and Venezuela: 3,713.4 km*°,

yielding an overall total of 6,665.9 km/.

Discussion

In cases where a species is widely distributed, and

especially when its range is disjunct, it 1s important to

assess whether significant intraspecific variation occurs.

Duellman and Gray (1983) reported no significant

morphological differences between the three respective

populations of Flectonotus fitzgeraldi from Trinidad,

Tobago, and Venezuela. The data here show highly

similar genetic divergence between populations, while

the Trinidad and Tobago animals were more closely

related between them than to those from Venezuela.

The lack of shared haplotypes suggests some degree of

genetic isolation among all three localities.

La Marca et al. (2004) assessed the conservation

status of F) fitzgeraldi as Endangered, on grounds of

its area of occurrence being less than 5,000 km7?, its

distribution being severely fragmented, and continuing

declines in the extent and quality of its habitat. However,

this assessment was not based on any published research.

In order to obtain field data, it was necessary to establish

an optimized survey methodology. These frogs call from

the leaves of host plants, which encompass phytotelmata

(bromeliads, Heliconia). Audio surveys at the time

just following sunset are the best survey method for

establishing presence or absence, either by walking, or

by the use of a frequently stopping vehicle which can

cover a longer distance at the critical time. With this

method, we confirmed the frog’s presence in the forests

of the northeastern half of Tobago, and found it to occur

wherever the host plants are common. In Trinidad, we

greatly increased the frog’s known occurrence to include

the whole of the Northern Range mountains and the full

length of the eastern half of the island; however, more

surveys are needed to establish its extent in southwestern

Trinidad. Our surveys in Venezuela have been less

extensive, but the frog’s presence was established further

west and south than previously recorded. The total

combined area of occurrence in the three countries is

6,665.9 km?.

The assessment of frog population size in the tropics

is highly problematic. Species such as F: fitzgeraldi are

widely distributed and may call throughout the year

Amphib. Reptile Conserv.

(Kenny 1969); they do not congregate to breed over a

short time at a few established ponds. Only males call, and

not necessarily every night. Greener et al. (2017) found

that day-time visual sampling of the Trinidad stream frog

underestimated the actual population (as established by

removal sampling) by a factor of about three. Similarly,

Lehtinen et al. (2016) showed, using new methods, that

the Tobago stream frog, Mannophryne olmonae, is more

abundant and widespread than previously considered.

Our population estimates for F) fitzgeraldi should not,

therefore, be regarded as true measures of the adult male

population, let alone the total numbers of all classes of

these frogs. However, we do think that our numbers

could be valuable in assessing trends, when compared

with future surveys using similar methods in the same

locations. The data from Trinidad’s Morne Bleu Ridge

should be particularly useful in the future for assessing

changes in a relatively undisturbed site.

In Venezuela, habitat loss in the Paria Peninsula

is not a new issue (Kaiser et al. 2015), having been

documented as early as the beginning of the 1940s.

Beard (1945) found that local people were responsible

for habitat disturbance mainly in the southern versant of

the mountains, and that human density appeared to be

too low to be in accordance with the vast area already

deforested at that time. A similar case occurs in La

Cerbatana, where a continued deforestation occurs below

500 m (maximum altitude 1,000 m), but to a lesser degree

than in Campeare, where natural vegetation only remains

on the summits of the mountains (ca. 950 m). Finally, the

natural forest formations present in the Turimiquire range

are subject to some strong threats, as observed during

a visit to the southeast side of the massif in 2006. The

mountain area between 300 m and 1,200 m 1s severely

affected by human activities. Pristine forests have been

lost while the landscape has shifted to agricultural spaces

and areas for human settlements. Land is mainly used

to cultivate tomato and coffee, and to a lesser extent for

livestock. Deforestation and cattle grazing have caused

erosion and loss of nutrients in the soils. Additionally,

these activities have encouraged the excessive use of

agrochemicals, mostly monophosphates, affecting the

mountain water bodies. Recently, however, most of

these crops appear abandoned. Tate (1935) mentioned

that the high elevations of Turimiquire did not escape

deterioration, and noticed important areas devoted to

coffee crops near 2,000 m elevation in the massif.

In conclusion, La Marca et al. (2004) estimated the

total extent of occurrence of F: fitzgeraldi to be less than

5,000 km? and assessed its population to be declining.

The low extent of occurrence, distribution in three

separate countries, and declines in extent and quality

of habitat led to a status assessment of Endangered. We

show here that the species’ extent of occurrence is well

over 6,000 km? (Supplementary Fig. S1), that there is no

evidence of population decline where we have been able

to measure it (Trinidad), and that the extent and quality

June 2021 | Volume 15 | Number 1 | e237

Flectonotus fitzgeraldi in Trinidad, Tobago, and Venezuela

of habitat in two countries (Trinidad and Tobago) show

no signs of decline. However, because there are threats to

habitat in the country with the largest area inhabited by

the species (Venezuela), it is probably best to regard F’

fitzgeraldi as Vulnerable.

Acknowledgements.—For allowing us to examine

specimens under his care, we are grateful to Edward

Camargo (EBRG). Gilson Rivas thanks Luis Sibira

for his valuable help in the field, and A.L. Viloria and

J.M. Gonzalez for their comments. Funding to work in

Paria, Venezuela, has been generously provided by The

Biodiversity Consultancy (Cambridge, United Kingdom)

through Oro Verde (Bonn, Germany) to Gilson Rivas and

Mayke De Freitas. Fieldwork in Trinidad and Tobago

by staff and students of the Universities of Glasgow,

Edinburgh, and Strathclyde was supported by small

grants from many institutions including the Universities,

the Glasgow Natural History Society, Dennis Curry’s

Charitable Trust, the Thriplow Trust, and the Gilchrist

Educational Trust. We thank all the students who

helped with the fieldwork. We also thank the members

of the Trinidad and Tobago Field Naturalists’ Club who

contributed observations to the annual Bioblitzes from

which we have drawn some of the data reported here.

Rick Lehtinen acknowledges the Tobago House of

Assembly for issuing research permits and the College

of Wooster (Wooster, Ohio, USA) for funding. We also

thank two reviewers (S. Lotzkat, H. Kaiser) for making

suggestions which allowed us to improve this paper.

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Flectonotus fitzgeraldi in Trinidad, Tobago, and Venezuela

Appendix 1. Specimens examined.

Flectonotus fitzgeraldi. VENEZUELA: Monagas: La Margarita, Turimiquire, 1,200 m, 10°10’32”N, 63°30’24”W

(EBRG 7034-35). Sucre: Mauraco, N del Pilar, 10°40°17,62”N, 63°06’38,64”"W (EBRG 519-520). Marauquito,

Peninsula de Paria, 400 m, (MHNLS 10859). Vertiente Sur de Cerro Humo, Peninsula de Paria, 800 m, 10°42’ N,

62°37’ W (MBLUZ 0395, 0448). Alrededores de las Melenas, Peninsula de Paria, 10°41°32.1”N, 62°37°24.9"W

(MHNLS 15741, 16187). Camino desde Macuro a los Chorros, Peninsula de Paria, 500 m, 10°41°54”N, 61°54’45”W

(MHNLS 16199-17201). Uquire, vertiente Norte de la Peninsula de Paria, 150 m, 10°43’N, 63°58’ W (EBRG 2585).

Cachipal, Peninsula de Paria, 10°38’02.5”N, 62°45’01.8’W (MBLUZ 447). Cerro La Cerbatana, ~800 m, 10°37’ N

-63°10’ W (EBRG 7336-7337). Cerro Campeare, 800 m, 10°32’45.4”N, 63°19°45.6”W (EBRG 7343-7346). Macuro,

Peninsula de Paria (photographed in Barrio-Amoréos et al. 2019: 54).

Jo Smith is a general zoology enthusiast, who accidentally ended up studying and falling in

love with treefrogs over the course of her Ph.D. on adhesion in the Hylidae, which involved

many enjoyable years going back and forth to Trinidad from Glasgow on field work. She is

now a Senior Lecturer at the Bangor University, Wales, where she tries her best to remind

their very keen zoology with herpetology students that snakes are not the only herps.

Michael J. Jowers is an evolutionary biologist with broad interests in the processes and the

timings of speciation. His work focuses on tropical island biogeography, phylogeography,

systematics, population genetics, taxonomy, and conservation. Michael is deeply involved

in amphibian and reptile studies from the islands of Trinidad and Tobago (Lesser Antilles),

but he is also interested in other organisms such as birds, mammals, and insects. He actively

leads studies throughout South America, Africa, Europe, and Asia.

Renoir J. Auguste is a Trinidad and Tobago herpetologist. Renoir received his M.Sc. in

Biodiversity Conservation from The University of the West Indies, St. Augustine Campus,

Trinidad and Tobago, and is interested in the ecology and conservation of amphibians and

reptiles. He has conducted herpetological surveys across Trinidad and Tobago for national

baseline surveys aimed at improving protected areas, as part of his academic degrees. He

also volunteers with the local environmental NGO Trinidad and Tobago Field Naturalists’

Club, in which he has held the position of President for three years.

Paul A. Hoskisson is Professor of Molecular Microbiology at the Strathclyde Institute of

Pharmacy and Biomedical Sciences (Scotland) and the Royal Academy of Engineering

Research Chair in Engineering Biology. While his research is primarily focused on

the biosynthesis of antibiotics, but he also maintains research interests in amphibian

reproduction, behavior, and conservation. Paul is a member of Royal Society of Biology

Council.

Amphib. Reptile Conserv. 48 June 2021 | Volume 15 | Number 1 | e237

Amphib. Reptile Conserv.

Smith et al.

Cammy Beyts is a Ph.D. student in Behavioral Ecology at The University of Edinburgh,

Scotland, within the Institute of Evolutionary Biology. Cammy’s research investigates the

effects of local adaption and the development environment on inter-individual differences

in tadpole behavior in species such as Engystomops pustulosus and Leptodactylus fuscus.

Cammy has been conducting her fieldwork in Trinidad since 2018, and regularly assists with

additional projects on a variety of herpetofauna while there.

Greig McInnes Muir is a recent Zoology graduate from the University of Edinburgh,

Scotland. Greig cultivated a passion for herpetology during his undergraduate research project

on the repeatable inter-individual differences in behavior of the model species Engystomops

pustulosus, while on expedition to Trinidad. Currently working within the pharmaceutical

industry, Greig 1s still a keen naturalist and is always looking for ways to keep connected to

nature, conservation, and behavioral ecology.

Mark Steven Greener developed an interest in amphibians and reptiles at a young age. He

further cultivated this passion at the University of Glasgow, Scotland, where he participated

in and led several scientific expeditions. Following this, Mark moved to the Wildlife Health

Ghent lab group at the University of Ghent, Belgium, pursuing a Ph.D. on the dynamics of

chytrid fungus in Northern Europe. He has been a part of multiple projects and has published

several papers on the amphibians and other wildlife of Trinidad.

Daniel G. Thornham is a zoologist and entomologist at the School of Natural Sciences,

Bangor University, North Wales, looking at relationships in phytotelm ecology, biomechanics,

and proximate behavioral ecology. Using pitcher plants (Nepenthaceae) and bromeliads

(Bromeliaceae) as model ecological systems, Dan explores the specializations of animals living

in the aerial aquatic habitats formed by these plants. In Bangor, Dan teaches undergraduate

research skills, invertebrate biology, and field techniques modules, and holds a number of

academic roles in the School. He has been visiting Trinidad periodically since 2000 and loves

nothing more than eating doubles after a long climb into the canopy to explore the bromeliads.

Isabel Byrne is an Irish researcher who received a B.Sc. in Zoology from the University

of Glasgow, Scotland. Her previous research focused on the herpetofauna of the rainforests

and coast of the North East of Tobago, with special interest in the Tobago Glass Frog,

Hyalinobatrachium orientale tobagoense. In 2020, she completed an M.Sc. in One Health:

Humans, Animals, and the Environment, at the Royal Veterinary College and the London

School of Hygiene and Tropical Medicine in England. She is currently working as a research

assistant at the London School of Hygiene and Tropical Medicine on the analysis of spatial and

environmental data related to malaria vector abundance, and epidemiological risk analysis.

49 June 2021 | Volume 15 | Number 1 | e237

Flectonotus fitzgeraldi in Trinidad, Tobago, and Venezuela

Richard (“Rick”) Lehtinen has studied the ecology, evolution, behavior, and conservation

of amphibians and reptiles for 25 years. He has authored or co-authored over 50 books,

monographs, and research articles on amphibians and reptiles with primary interests in

the specializations of plant-breeding frogs, the evolution of parental care behavior, and

the long-term study of population dynamics. Primarily a field biologist, Rick has enjoyed

probing the secrets of nature in Costa Rica, Madagascar, Taiwan, Trinidad and Tobago,

and the United States. He holds a Ph.D. from the University of Michigan and is currently

Professor of Biology at the College of Wooster (Ohio, USA).

Meredith Eyre worked as a student in Dr. Lehtinen’s group at the College of Wooster

(Ohio, USA), and under his guidance, her senior thesis explored the natural history and

habitat selection of Flectonotus fitzgeraldi in Tobago. She continues to be inspired by the

research opportunities created for her at the undergraduate level, and after completing her

M.S. at the Ohio State University, she has dedicated herself to the development of similarly

empowering opportunities for others. She currently serves as a Laboratory Instructor at

Carlow University (Pennsylvania, USA), where she delights in working primarily with

introductory-level students as they begin their journey into the field of biology.

Michael G. Rutherford is a naturalist based in Scotland. A lifelong interest in animals

led him to a degree in Zoology from Glasgow University, Scotland, followed by a mas-

ters in conservation biology at James Cook University, Australia. His jobs have included

research assistant studying skinks in Queensland, tropical house zookeeper for Newquay

Zoo (England), curator of invertebrates for Glasgow Museums (Scotland) and curator

of the University of the West Indies Zoology Museum in Trinidad and Tobago. His re-

search interests include reptiles, land snails, millipedes, oilbirds, and biological recording.

John C. Murphy is a naturalist with a focus on snakes. When he is not hiking in the

desert or examining specimens in the lab, he is often writing about reptiles. Murphy is a

retired science educator who got serious about his lifelong fascination with lizards and

snakes in the early 1980s when he and his family made their first trip to Trinidad. The

work on Trinidad and Tobago provided valuable lessons that shaped his views of nature

and evolution, and today he is still working on the eastern Caribbean herpetofauna. In

the 1990s, he did some work on homalopsid snakes in Southeast Asia with others from

the Field Museum (Chicago, Illinois, USA). He now resides in southeastern Arizona and

is involved in multiple projects on arid habitats and the impacts of climate change on

biodiversity. His most recent book is Giant Snakes, a Natural History (with co-author

Tom Crutchfield). Born and raised in Joliet, Illinois, he first learned about reptiles on his

grandfather’s farm by watching Eastern Garter Snakes emerge from their winter dens and

Snapping Turtles depositing their eggs at the edge of a cattail marsh.

Mayke De Freitas Santos is a conservationist with postgraduate studies in International

Environmental Law (SOAS) and Conservation Leadership (University of Cambridge,

England) who has worked for various international NGOs both in Venezuela and Europe.

Mayke focuses on the governance of protected areas, as well as remote sensing and habitat

quality assessments of amphibian species. He is a contributor to the IUCN Red List for

several species of amphibians and reptiles from the Peninsula de Paria, Venezuela, where

he has worked since 2011.

Amphib. Reptile Conserv. 50 June 2021 | Volume 15 | Number 1 | e237

Smith et al.

Gilson A. Rivas was born in Caracas, Venezuela. He currently serves as co-editor of the

scientific journal Anartia, and is acollection manager at the Museo de Biologia de la Universidad

del Zulia, Maracaibo—a Venezuelan centennial university that began academic activities on

11 September 1891. For over two decades, Gilson has been devoted to the taxonomy and

conservation of the neotropical herpetofauna, having authored or co-authored more than 100

academic publications, and describing over 30 new species of amphibians and reptiles, and a

new genus of dipsadine snake, P/esiodipsas. Gilson is the author (with G. Ugueto) of the book

Amphibians and Reptiles of Margarita, Coche, and Cubagua; and together with M. De Freitas,

H. Kaiser, C.L. Barrio-Amoros, and T.R. Barros produced Amphibians of the Peninsula de

Paria: a Pocket Field Guide. Gilson’s research interests are focused on the herpetofauna of the

Venezuelan coastal range and insular ecosystems, as well as the influences of invasive species

and human development and their impact on the native fauna.

J. Roger Downie is a semi-retired Professor of Zoological Education at the University of

Glasgow, Scotland. During his long association with Trinidad and Tobago, he has contributed

to research on amphibians and marine turtles in collaboration with Glasgow students, and

many others, on numerous research expeditions.

Supplementary Fig. S1 Maps showing the GPS coordinates where Flectonotus fitzgeraldi was found to be present

and clipped polygon shapes used to calculate the total area of occurrence for F’ fitzgeraldi in Trinidad, Tobago, and

Venezuela (Supplementary Fig. Sla—c). The areas of occurrence are Trinidad: 2,922.4 km’; Tobago: 30.1 km; and

Venezuela: 3,713.4 km’.

| F fitzgeraldi

Fig. Sla. Presence and total area of Flectonotus fitzgeraldi occurrence in Trinidad.

Amphib. Reptile Conserv.

oA June 2021 | Volume 15 | Number 1 | e237

Flectonotus fitzgeraldi in Trinidad, Tobago, and Venezuela

Fig. Slb. Presence and total area of Flectonotus fitzgeraldi occurrence in Tobago.

Fig. Sle. Presence and total area of Flectonotus fitzgeraldi occurrence in Venezuela.

Amphib. Reptile Conserv. 52 June 2021 | Volume 15 | Number 1 | e237

Smith et al.

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June 2021 | Volume 15 | Number 1 | e237

Amphib. Reptile Conserv.

Flectonotus fitzgeraldi in Trinidad, Tobago, and Venezuela

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Amphib. Reptile Conserv.

Smith et al.

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June 2021 | Volume 15 | Number 1 | e237

Amphib. Reptile Conserv.

Flectonotus fitzgeraldi in Trinidad, Tobago, and Venezuela

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Amphib. Reptile Conserv.

Official journal website:

amphibian-reptile-conservation.org

Amphibian & Reptile Conservation

15(1) [General Section]: 57-70 (e274).

Can artificial retreat sites help frogs recover after severe

habitat devastation? Insights on the use of “coqui houses”

after Hurricane Maria in Puerto Rico

*Patricia A. Burrowes, ‘Abner D. Hernandez-Figueroa, ‘Gustavo D. Acevedo,

‘Junangel Aleman-Rios, and ?Ana V. Longo

'Department of Biology, P.O. Box 23360, University of Puerto Rico, San Juan, Puerto Rico 00931 *Department of Biology, P.O. Box 118525,

University of Florida, Gainesville, Florida 32605 USA

Abstract.—On September 2017, Hurricane Maria swept over Puerto Rico as a Category 4 storm. Severe

canopy loss, augmentation of forest floor debris, and a significant increase in temperature and light reaching

the understory were among the most evident changes at El Yunque National Forest, where a population of

Eleutherodactylus coqui frogs has been monitored over the past 30 years. When sampling was re-established,

the frogs could be heard calling, but it was very difficult to find them among the complexity of vegetation

in the forest floor. We inferred that canopy disturbance had left frogs without optimal arboreal habitats for

retreat, nocturnal perching, feeding, and reproductive activities, and wondered whether they would use

artificial habitats placed in the forest understory. To test this, two types of artificial habitats (i.e., ““coqui houses”)

were introduced in the forest understory, consisting of either open PVC pipes or single-entrance natural bamboo

shoots. Surveys were conducted twice a month for 15 months in an experimental transect with coqui houses, and

a control transect without them. Data were collected on the occupancy rate of the artificial sites, type of usage,

time of day occupied, and the number of E. coqui observed. The effects of time since the hurricane, microhabitat

temperature, type of coqui house, and seasonality on the occupancy rate were also evaluated. Results showed

that coquis used bamboo houses mostly during daytime as retreat and nesting sites, whereas the PVC houses

were used mostly at night as calling sites. Daytime occupancy of coqui houses showed a significant bell-shaped

pattern over time since the hurricane. This may be explained by a steady increase in usage after severe forest

damage, a peak during the stressful cool-dry season, and a decline afterwards as the forest began to recover. No

differences were found in frog counts between experimental and control transects, probably because the coquis

could also hide among the fallen vegetation, but either disparities in forest conditions or inappropriateness of the

methods for estimating population numbers may have overshadowed this effect. Coquis used artificial houses

more often during the most stressful environmental conditions, suggesting that these shelters may serve to

enhance habitat quality for amphibians after extreme weather events.

Keywords. Amphibians, conservation, E/eutherodactylus, extreme events, habitat augmentation, retreat sites

Citation: Burrowes PA, Hernandez-Figueroa AD, Acevedo GD, Aleman-Rios J, Longo AV. 2021. Can artificial retreat sites help frogs recover after

severe habitat devastation? Insights on the use of “coqui houses” after Hurricane Maria in Puerto Rico. Amphibian & Reptile Conservation 15(1)

[General Section]: 57-70 (e274).

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: 14 June 2021

Introduction

Trends of recent climate change (1961—2010) in the Ca-

ribbean region show evidence of an increase in the fre-

quencies of warm days, warm nights, and extreme high

temperatures with a corresponding decrease of their

cooler equivalents (Stephenson et al. 2014). An increase

in minimum temperatures is significant at regional (Pe-

terson et al. 2002; Stephenson et al. 2014) and local

scales, such as the Puerto Rican eastern highland forests

(Burrowes et al. 2004). While precipitation patterns for

Correspondence. *“pburrowesupr@gmail.com

Amphib. Reptile Conserv.

the Caribbean are not as clear, some studies have shown

a significant drying trend (Nurse and Sem 2001; Neelin

et al. 2006). In addition, the frequency of extreme events,

like hard rains and hurricanes, has increased significantly

in the last decade, and the International Panel for Cli-

mate Change (IPCC) anticipates further increases in the

future (Peterson et al. 2002; IPCC 2014, 2018; Oppen-

heimer et al. 2019). Due to factors intrinsic to the biology

and physiology of amphibians, these patterns of climate

change represent a threat to these animals. For example,

many tropical ectotherms are thermoconformers, having

June 2021 | Volume 15 | Number 1 | e274

Coqui frogs and Hurricane Maria

-65.79° -65.785°

°o

ro]

ico. ;

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* - Palo Colorado, El Yunque

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a reduced ability to thermoregulate compared to other

vertebrates adapted to the climatic heterogeneity of high-

er latitudes (Navas 1996a,b; Huey et al. 2009). For the

anurans in the genus E/eutherodactylus, which represent

an important component of the endemic vertebrate fauna

of the Caribbean (Hedges 1999; Duellman 1999; Joglar

2005), this may pose an additional challenge because

amphibians lack epidermal protection that can reduce

evapotranspiration rates during hot days. Thus, global

climate change patterns for the Caribbean pose a risk for

the Eleutherodactylus of Puerto Rico (locally known as

“coquis”), which are already dealing with other threats

such as the pathogenic chytrid fungus (Batrachochytrium

dendrobatidis Longcore, Pessier and Nichols, 1999) that

causes chytridiomycosis (Burrowes et al. 2004, 2008a,b,

2017; Longo and Burrowes 2010; Longo et al. 2013).

Extreme atmospheric events, such as strong hurri-

canes, can affect coqui populations by changing local

temperature regimes and habitat structure. A study on

the effect of Hurricane Hugo (Category 3 in 1998) on

a population of Eleutherodactylus coqui at the Luquillo

Experimental Forest (LEF) of Puerto Rico, revealed an

increase in adults, albeit at a smaller body size, and a

decrease in juveniles (Woolbright 1991, 1996). The pro-

liferation of adults was attributed to a greater availability

of retreat sites among the fallen debris and a decrease in

invertebrate predators, while the reduction of juveniles

was explained by a decrease in reproductive activity due

to changes in microclimate (Woolbright 1991). Another

study experimentally evaluated the effect of two conse-

quences of hurricanes, canopy disturbance and increased

Amphib. Reptile Conserv.

-65.78°

-65.775° -65.77°

Study Area

O21 400m |

Fig. 1. Map showing the location of El Yunque National Forest in Puerto Rico, and the location of the study transects.

deposition of debris in the forest floor, on a population of

coquis in the LEF in Puerto Rico (Klawinski et al. 2014).

In contrast to the results of Woolbright (1991), this study

found that while canopy disturbances were detrimental to

E. coqui abundance in experimental plots, an increase in

forest debris had no effect (Klawinski et al. 2014).

In September 2017, Hurricane Maria hit Puerto Rico

as a strong Category 4 storm with maximum sustained

winds of 155 mph (~250 km/hr). High energy winds

spread at 280 km/hr over the island along with heavy

rainfall resulting in catastrophic flooding in many areas

(NOAA 2017). As a consequence, critical damage to the

island’s infrastructure occurred and Puerto Rico was de-

clared a disaster area by the Federal Emergency Man-

agement Agency (FEMA). Natural ecosystems were also

severely affected. At El Yunque National Forest in the

eastern highlands of the island (Fig. 1), most large trees

were defoliated and lost medium to large branches, while

others were snapped or uprooted by Hurricane Maria’s

strong winds. A large amount of vegetative debris accu-

mulated on the forest floor causing an increase in struc-

tural complexity at that level. These changes and other

complex consequences of hurricanes to ecosystems were

discussed by Lugo (2008). Considering the negative ef-

fects of Hurricane Hugo on E. coqui populations (Wool-

bright 1991; Klawinsky et al. 2014), and with the knowl-

edge that Hurricane Maria had been much stronger, we

were concerned about the response of the population of

coqui frogs that we have been monitoring for over 30

years at higher elevations in El Yunque National forest

(Burrowes et al. 2004; Longo and Burrowes 2010).

June 2021 | Volume 15 | Number 1 | e274

Burrowes et al.

We obtained permission from the U.S. Forest Service

to evaluate our working area at El Yunque in December

2017, but due to security measures, we were not allowed

to conduct nocturnal work in the forest until March 2018.

At that time, finding our transects and clearing a path

through forest debris to re-establish amphibian moni-

toring was arduous work. Initial surveys of the coqui

population revealed that male frogs were still calling,

but adults were difficult to observe because they were

not active in their typical understory nocturnal sites, and

juveniles, common in pre-hurricane surveys, were un-

detected. This was probably due to the absence of arbo-

real vegetation such as bromeliads and palm-frond axils

which fell with the hurricane winds, as well as the loss

of moss-covered tree trunks and branches that were con-

cealed by leaves overhanging from the canopy before the

hurricane, providing ideal hiding places for active frogs.

Thus, the objective of this study was to provide artificial

habitat in the form of “coqui houses” in the forest under-

story and quantify how much the frogs would use them

for their regular activities, as well as the factors that may

be associated with their usage. The data obtained could

also serve to evaluate the potential of this strategy as a

mitigation tool in the face of severe forest devastation.

The placement of artificial habitats has been used to

increase reproductive activity and success of other ver-

tebrate species of conservation concern. For example,

Grubb and Bronson (1995) used artificial nesting sites

made of PVC tubes to increase reproductive activity and

success of Passeriformes birds (chickadees) in Ohio,

USA. They found that when using artificial nesting sites,

the survival of juveniles of one species of chickadees

increased to 100%, and that birds used artificial nesting

sites even in areas where natural nesting sites were abun-

dant (Grubb and Bronson 1995). In Baja California, os-

preys were observed using artificial nesting sites within

a development area, resulting in an increase in reproduc-

tive activity and population size (Ortega-Rubio 1995).

For the European Storm Petrels, for which the abundance

of nesting sites 1s considered a limiting factor, the place-

ment of artificial nesting sites proved to be effective at

increasing population growth and density (De Leon and

Minguez 2003). Among amphibians, a study in Australia

showed that water dams in farmlands supported a similar

number of species as natural ponds, and highlighted the

role of well-designed artificial habitats in frog conser-

vation (Hazell et al. 2004). In Puerto Rico, Stewart and

Pough (1983) placed artificial nest houses constructed

from bamboo shoots in selected plots at “El Verde” in the

LEF (350 m asl), and studied their effect on a population

of Eleutherodactylus coqui. The authors found that the

bamboo houses were often used as retreat and nesting

sites, and resulted 1n significant increases in reproductive

activity and population numbers. They concluded that re-

treat sites were a limiting factor for E. coqui population

abundance (Stewart and Pough 1983).

Based on the positive results obtained by Stewart and

Amphib. Reptile Conserv.

Pough (1983), we expected that coqui frogs confront-

ing the habitat devastation caused by Hurricane Maria

would use artificial habitats placed on trees in the forest

understory as retreat, calling, perching, mating, and nest-

ing sites. If so, we hypothesized a consequent increase

in nocturnal frogs counts in experimental versus control

transects because the coqui houses would encourage re-

productive activity. The factors that influenced occupan-

cy rate and the type of uses given to different types of ar-

tificial coqui houses were also investigated in an attempt

to determine their potential effectiveness for mitigating

the effects of habitat loss. Understanding the patterns of

species responses to dramatic changes in habitat is 1m-

portant in order to develop effective conservation prac-

tices (Gascon 1993). The results of this work may guide

future management strategies to help the recuperation of

amphibians and/or other species after severe forest dam-

age due to extreme climatic events.

Materials and Methods

Study Area

This work was conducted at El Yunque National Forest

at 661 m asl in the Luquillo mountains of Puerto Rico.

The study transects were located within the Palo Colo-

rado Forest formation (18°1875.8"N, 65°47’7.4°W; Fig.

1), characterized by the tree with the same name, Cy-

rilla racemiflora, as well as the Sierra Palm, Prestoea

montana (Ewel and Whitmore 1973; Harris et al. 2012).

Other species, such as Micropholis garcinifolia (Caim-

itillo Verde), Calycogonium squamulosum (Jusillo), and

Croton poecilanthus (Sabinon), are also common trees

in the Palo Colorado Forest, and epiphytic bromeliads

in the genus Guzmania, which are an important habitat

for Eleutherodactylus frogs (Joglar 1998), are abundant

(Stephenson et al. 1999; Harris et al. 2012). The average

minimum temperature (nocturnal) and daily precipitation

at the Palo Colorado Forest are 21.5 °C and 9.9 mm, re-

spectively, but vary with the marked seasonality of Puer-

to Rico—being cooler and drier from January to April

and warmer and wetter from May to December (Longo

et al. 2010). Three species of Eleutherodctylus, E. coqui,

E. hedricki, and E. portoricensis, can be found within the

transect in the Palo Colorado forest. Following Hurricane

Maria in 2017, the transect sites within this forest exhib-

ited considerable canopy cover change. Table 1 shows an

attempt to broadly categorize the stages of forest dam-

age and chronological recuperation based on field notes

and photographs (Fig. 2). The initial damage level was

approximately 90-99% devastation, with a large propor-

tion of that being vegetation accumulated in the forest

floor, which then progressed to a considerable recupera-

tion of up to 50% of the canopy cover by the end of 2019.

The chronology presented in Table 1 may seem rather

rapid, but research has shown that forest recovery after

hurricane damage is faster in the tropics than in temper-

June 2021 | Volume 15 | Number 1 | e274

Coqui frogs and Hurricane Maria

Fig. 2. Change in forest structure in the Palo Colorado forest transect (El Yunque) due to Hurricane Maria and corresponding

damage/recovery stages according to Table 1. (A) Before the hurricane. (B) Same site after the hurricane, stage 1. (C) Moderate

recuperation, stage 3. (D-E) Canopy dominated by Sierra Palm fronds showing signs of further recuperation of original understory

vegetation, stage 4.

ate zones (Canham et al. 2010). In Puerto Rico, forests | Study Species

tend to recover basic structure within 10 to 20 years after

hurricane blowdowns (Walker 1991; Lugo and Helmer The biology and ecology of E. coqui (Fig. 3) was exten-

2004), and may achieve a tree size and density similarto sively reviewed by Joglar (1998); thus, only some details

those of primary forests after 40 years of recovery (Aide _ that are pertinent to this study will be highlighted here.

et al. 1996). The species is native to Puerto Rico where it is widely

Table 1. Stages of forest changes describing the timeline of hurricane damage and gradual recovery. These stages are based on

personal observations and field notes during daytime monitoring of coqui-houses at the Palo Colorado Forest (661 m) in El Yunque,

Puerto Rico.

Stages of hurricane

damage Forest description Approximate time-frame

(1 = greatest)

cao 7 a

90-99% of the forest canopy gone; remaining standing trees 90 Hurricane Maria, Sep

i ee é

1 95% defoliated; vegetation fallen from the canopy accumulated in 2017-Sep 2018

the floor

85-90% of the forest canopy gone; palm trees grow back their

fronds; canopy vegetation decomposing and understory vegetation Oct 2018—Jan 2019

2 growth dominated by herbaceous vegetation and opportunistic fast-

growing plants that tolerate sunlight; recuperation of standing trees

very limited

70-85% of forest canopy gone; standing trees growing leaves on

3 branches, and native understory vegetation becomes apparent with

new saplings restricting light and limiting the growth of herbs and

opportunistic plants

Feb 2019-May 2019

50-70% of forest canopy gone; standing trees recuperating leaves

4 to about 70%, and native understory vegetation growing with new

saplings at about 5 m high; forest understory clear of herbs and other

light-loving plants; canopy dominated by sierra palm fronds

Jun 2019-Dec 2019

Amphib. Reptile Conserv. 60 June 2021 | Volume 15 | Number 1 | e274

Burrowes et al.

distributed from urbanized to pristine areas, and from sea

level to highland wet forests (O—-1,189 m asl). It is an im-

portant nocturnal predator which consumes a variety of

prey, predominantly arthropods (Woolbright 1991). The

size of E. coqui varies considerably with the elevation,

with individuals from higher elevations being the largest

(Narins and Smith 1986). At the study site (661 m asl), the

mean size of adult females measured as the snout-to-vent

length (SVL) is 46.4 mm and that of adult males is 36.4

mm (Joglar 1998). Similar to other Eleutherodactylus

species in Puerto Rico, E. coqui females are larger, pre-

sumably due to the cost of producing clutches of heavily-

yolked, direct-developing eggs (Townsend and Stewart

1986; Woolbright and Stewart 1987). The common coqui

uses various substrates as daytime retreats, including leaf

litter, rock cavities, palm axils, and bromeliads. As nest-

ing sites, they may use arboreal bromeliad axils, cracks

within the bark of tree trunks, curled palms fronds, or

large Cecropia leaves. It is common to observe males

calling at night from vegetation 1—3 m above the ground.

Reproductive activity is present all year around, but it is

more frequent during the warm-wet season (Woolbright

and Stewart 1987; Joglar 1998). During mating encoun-

ters, males call to attract females to a protected nesting

site where amplexus takes place. Mating behavior usu-

ally lasts between 8.5—12 hours, and in contrast to most

anurans, eggs are fertilized internally (Townsend et al.

1981). Males offer parental care to the clutch, which can

last from 16 days in the higher temperatures typical of

Amphib. Reptile Conserv.

Fig. 3. Eleutherodactylus coqui perching on natural habitat. (A) Female on branch. (B) Male on tree trunk.

lowlands, or up to 27 days in cooler, highland tempera-

tures (Townsend and Stewart 1986).

Field Work

For this study, two 50 m x 3 m transects were sampled, an

experimental transect where coqui houses were placed,

and a control left unmanaged, approximately every two

weeks for 15 months from August 2018 to October 2019.

These transects are located only 100 m apart at the study

site and have been monitored over many years to esti-

mate population fluctuations and study the responses of

coqui frogs to chytridiomycosis (Burrowes et al. 2004,

2017; Longo et al. 2010; Longo and Burrowes 2010). Vi-

sual encounter surveys were used to count the number

of adult frogs active from the middle of the transects to

a distance of 1.5 m at each side of the forest (Heyer et

al. 2014) while also monitoring coqui houses for occu-

pancy. Both transects were sampled simultaneously start-

ing shortly after dusk (1830-1900 h) for approximately

2.5 hours by two or three trained people. This time range

was chosen because it is the peak time of activity for E.

coqui (Drewry 1970; Woolbright 1985). If a frog was

found in one of the coqui houses, its behavior was re-

corded to infer the particular uses made of the artificial

habitats, such as for retreat, calling, mating, or nesting

and parental care. To determine potential differences in

occupancy of coqui houses during daytime versus night-

time, coqui houses were checked twice, first at approxi-

June 2021 | Volume 15 | Number 1 | e274

Coqui frogs and Hurricane Maria

mately 1600 h and then at night during the regular survey

as described above. Juvenile frogs (SVL 5—18 mm) were

not considered for this study because debris and invasive

plant vegetation heavily covered the forest floor and the

lower vegetation where young coquis are active at night

(Burrowes et al. 2017). Thus, it was impossible to say

with certainty whether juveniles truly decreased, or if

their absence was an artifact of the difficulty in sampling.

Two different kinds of “coqui houses” (Fig. 4) were

used: (A) bamboo houses made of natural bamboo

cylinders cut at a node to provide a floor and a small

Opening carved into the lower part of the cylinder as a

single entrance point (Stewart and Pough 1983); and (B)

open-ended, hollow, white PVC pipes. The length of the

cylinders for both kinds of houses was 6.4 cm, and the

diameter did not exceed 4 cm. The top of the bamboo

houses was closed with strong duct tape such that frogs

could not get in or out, but they could be easily checked

for occupancy by peeling the tape back (Fig. 4). In the

experimental transect, a total of 22 bamboo houses and 22

PVC houses were deployed. One of each type was placed

every 5 m in the understory, tied to trees or branches

1.5—2.0 m above ground in the forest understory (Fig. 4).

Similar types of coqui houses are used to trap invasive E.

coqui in Hawaii (Control of Coqui Frogs in Hawaii 2008;

Pitt et al. 2012).

In order to document the changes in temperature as-

sociated with forest cover loss after Hurricane Maria

and the structural damage caused to the forest canopy,

HOBO® data-loggers were used to monitor ambient

air temperature in the understory of the transects dur-

Amphib. Reptile Conserv.

Fig. 4. Artificial coqui houses built with (A) bamboo shoots and (B) PVC tubes.

ing the 15 months of this study. Because microhabitat

temperatures are routinely monitored with dataloggers

for other studies in these transects, one datalogger was

deployed during the hurricane that allowed us to track

the temperature changes during the storm itself (Fig. 5A).

—-~

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a < ai 8 2 a

3 a a 7 vt =

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Sep2015 Sep2017 Sep2015 Sep2017

Fig. 5. Drastic changes in temperature at the transects in the

Palo Colorado forest of El Yunque as a consequence of Hur-

ricane Maria. (A) Ambient temperatures registered by HOBO

data logger in the forest understory before, during, and shortly

after Hurricane Maria. (B—C) Box plots showing variation in

forest microhabitat temperature by day and at night during the

month of September in 2015 (a non-hurricane year), and in

2017, the year that Hurricane Maria hit Puerto Rico.

June 2021 | Volume 15 | Number 1 | e274

Burrowes et al.

Q A

” o

g 3 § G Warm wet season i]

= — g B cool dry season

o i i io of ti

5s 8 hea

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Fig. 6. Variation in operative temperatures measured by frog

agar models in typical forest microhabitats after Hurricane Ma-

ria, showing a significant decrease during the cool-dry season

(in blue) during midday (A), and nighttime (B).

The HOBO® data-loggers were also used in the bamboo

coqui houses to record microhabitat temperature in the

forest understory during the duration of this study. The

thermal coupler was inserted into agar models designed

to mimic coqui frogs (Rowley and Alford 2010; Bur-

rowes et al. 2020) such that the microhabitat tempera-

tures would reveal the operative temperatures that frogs

would encounter at these sites (1.e., equilibrium body

temperatures that the animal experiences in its habitat

(Bakken and Gates 1975)). Temperature readings taken

in the same manner in this transect during the year 2015

allowed the assessment of changes tn the specific months

before and after Hurricane Maria.

Analyses

Bar graphs are used to illustrate the frequency of usage

of coqui houses by type and time of day through the sam-

pling surveys or time since the hurricane, and boxplots

are used to illustrate changes in temperature patterns. A

Generalized Mixed Linear Model (GLM) was applied to

evaluate factors that best predicted occupancy rates of

the artificial habitats. In the model, date of sampling and

coqui house number were considered random effects,

and time (days) since Hurricane Maria, microhabitat

temperature (mean of five days: four days immediately

before and sampling day), and the type of coqui house

(PVC versus bamboo) as fixed effects. Mann-Whiney

or ¢-tests were conducted to test for differences in tem-

perature in forest habitat that may be attributed to post

hurricane damage, and ANOVA was used to determine

whether mean frog counts were associated with transects

where coqui houses were placed and whether differences

in mean microhabitat temperatures were associated with

seasons. Chi-square was used to test for independence

Amphib. Reptile Conserv.

22 23

Temperature (C*)

19 20 21

Temperature (C*)

19 20 21

Feb 2015

Feb 2019

Feb 2015

Feb 2019

Year Year

Fig. 7. Box plots showing variation in forest microhabitat tem-

perature by day (A) and night (B) during the cool-dry season

(February) of 2015 (a non-hurricane year), and in 2019, 17

months after Hurricane Maria hit Puerto Rico.

on the type of coqui house used by day versus night, as

well as particular behaviors. Finally, a quadratic regres-

sion model was fitted to evaluate the predictive effect of

time since the hurricane on the daytime usage of coqui

houses. Statistical analyses were performed with Minitab

Release 1.5.3 (https://www.minitab.com/) and R statisti-

cal package (http://www.R-project.org/).

Results

Temperature Changes in the Forest

As an immediate result of canopy cover loss due to Hur-

ricane Maria, a drastic increase in the ambient tempera-

ture was registered by a data logger left in the forest until

its battery was drained shortly after (Fig. SA). In the mi-

crohabitats, a significant increase and high variability in

temperature was found when comparing a non-hurricane

year (September 2015) with that of September 2017, the

year of Hurricane Maria (Fig. 5B—C). This pattern was

evident both at mid-day when frogs are in retreat sites

(W = 46,697, P < 0.001), and at night when they are ac-

tive (W = 310,949, P < 0.001) (Fig. 5B—C). Temperature

data taken during this study showed that after Hurricane

Maria, frogs confronted operative temperatures spanning

22-28 °C during midday, and 18—24 °C at night in for-

est microhabitats (Fig. 6A—B). These temperatures are

alarming because they were measured at retreat sites

where frogs are protected from direct sunlight, and yet,

are on average 2 °C higher than air temperatures regis-

tered in a previous study during 2005—2007 in the same

forest transects (Longo et al. 2010). As expected, micro-

habitat temperatures post-Maria were significatively as-

sociated with seasonal changes, where higher tempera-

tures corresponded to the warm-wet season and lower

temperatures to the cool-dry season (F' = 79.47, df = 13,

P <0.001; Fig. 6A—B). A comparison of the mean opera-

tive temperatures recorded in the same manner in Febru-

ary 2015 during the cool-dry season of a non-hurricane

year, with those of February 2019 (17 months after Hur-

ricane Maria), showed a significant increase in the micro-

habitat temperature during the day (x = 1.08, T=-5.34, df

= 300, P < 0.0001) and at night (x = 0.56, T= -11.09, df

= 368, P < 0.0001) (Fig. 7A—B).

June 2021 | Volume 15 | Number 1 | e274

Coqui frogs and Hurricane Maria

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and by nighttime (B) surveys. The shaded area in (A) denotes sampling in months during the cool-dry season.

Coqui House Occupancy Rates

The results of a general mixed model revealed that (1)

time since the hurricane as a proxy of forest recovery,

(2) microhabitat temperature, (3) house type, and (4) the

interaction between time since the hurricane and micro-

habitat temperature were all significant predictors of oc-

cupancy rate of artificial coqui houses by E. coqui (Table

2).

Over the study period frogs that used these artificial

habitats were found more often during the day with an

occupancy rate up to 27.3% (mean 10.64, SD + 4.23),

and somewhat less during the night with a maximum oc-

cupancy of 13.6% (mean 4.94, SD + 2.20) during any

sampling event (Fig. 8). The time at which frogs oc-

cupied one of these artificial habitats was not indepen-

dent of the type of coqui house (X? = 104.05, df= 1, P<

0.001). Bamboo houses were used mostly during the day,

as retreat and nesting sites, while PVC houses were used

mostly at night for perching and/or calling (X? = 115.94,

df= 1, P< 0.001, Fig. 9).

The number of frogs occupying coqui houses dur-

ing the day was found to be inversely related to the

mean midday (1200-1600 h) temperature at the mi-

crohabitats (Y = -0.68x + 18.35, P = 0.0185 and R?=

37.13). Accordingly, frogs occupied bamboo coqui

houses significantly more often in the cooler dry sea-

son (shaded box) than in the warmer wet months (F' =

Table 2. Results of Generalized Mixed Linear Model (GLM) to determine the effects of several factors on occupancy of the artificial

coqui houses. The number of asterisks (*) indicates the level of statistical significance (a = 0.05).

Fixed effects Estimate Standard error Z value P lz|)

Intercept -38.21 6.549 -5.835 0.000 Fee

Time (days since hurricane) 0.068 0.0097 6.937 0.000 RK

Temperature 1525 O277 5.496 0.000 aa chs

House type PVC -1.323 0.519 -2.546 0.011 é

Time: temperature interaction -0.003 0.0004 -7.221 0.000 ee

Amphib. Reptile Conserv.

64 June 2021 | Volume 15 | Number 1 | e274

Burrowes et al.

Fig. 9. Different uses ascribed to the two types of artificial habitats (=coqui houses) placed in the forest. (A) Coqui frog using bam-

boo house as retreat site during the day. (B) Bamboo house used as nesting site with a double clutch. Note that eggs are observed

but the guarding male jumped away as the photo was taken. (C) PVC house used by a coqui as a nocturnal perching site. (D) PVC

house used by a coqui as a calling site during the night.

5.90, df = 13, P = 0.0258, Fig. 8A). The frequency of

daytime usage (i.e., occupancy rate) of coqui houses

through time since the hurricane, which serves as a

proxy of forest recovery (Table 1), fits a bell-shaped

curve that was explained by a quadratic model (y =

-0.0004x? + 0.426x — 102.24, P = 0.0026 and R? =

0.40, Fig. 8A). No pattern in the nocturnal usage could

be associated with time after the hurricane; nonethe-

less, the coqui houses (especially PVC) were used

consistently throughout the study (Fig. 8B). On four

occasions, bamboo houses were used for nesting/pa-

rental care, and during this period the males stayed

inside day-and-night for approximately 27 days until

the eggs hatched and the juveniles dispersed (Fig. 9).

Amphib. Reptile Conserv.

Contrary to what we expected, the mean number

of adult frogs observed per survey night was slightly

greater in the control transect (18.43 + 5.98) than in the

experimental transect (17.93 + 5.54; Paired 7= -0.313,

P= 0.7599; Fig. 9). The higher variance associated with

the abundance of frogs in the control transect may re-

flect greater habitat heterogeneity in this part of the forest

(Fig. 10).

Discussion

Hurricanes have many effects on forest ecosystems,

ranging from the physical damage to the forest and its

functional consequences to biodiversity and ecosystem

June 2021 | Volume 15 | Number 1 | e274

Coqui frogs and Hurricane Maria

Relative Abundance

Experimental Control

Fig. 10. Comparison of the relative abundance, measured as

the number of adult E/eutherodactylus coqui observed per sam-

pling night in the experimental transect where artificial coqui

houses were made available, versus the control.

processes, to the opportunity for evolutionary change

(Lugo 2008). Herein the thermal and structural chang-

es that Hurricane Maria caused to a highland forest in

Puerto Rico are documented, and results are presented on

the responses of coqui frogs to the placement of artificial

coqui houses on trees in the forest understory from which

their traditional habitat had been displaced.

The drastic effect of Hurricane Maria’s blowdown

over the study site at El Yunque was evidenced by a 6 °C

increase in ambient temperature in September 2017, right

after 1t passed over Puerto Rico (Fig. 5A). A comparison

of microhabitat temperatures in the months of September

2015, 17 years since the previous hurricane (Georges 1n

1998) and September 2017, show a significant increase

in temperature attributable to the loss of forest canopy

caused by Hurricane Maria (Fig. 5B—C). A similar com-

parison of microhabitat temperatures in the months of

February in 2015 and 2019 (Fig. 7A—B), revealed that 17

months after the hurricane, when the forest is considered

to be on recovery stage 3 (Table 1), frogs in the under-

story still confronted operational body temperatures ap-

proximately 1.0 °C higher than tn non-hurricane years.

The observed increase in temperatures by day and night

associated with the loss of canopy are expected to bring

about an increase in wind speed and a corresponding de-

crease in relative humidity within the forest (Tanner et

al. 1991; Lugo 2008). These thermal changes may ex-

acerbate the already stressful environmental conditions

for coqui frogs as their operative body temperatures rise

and the risk of dehydration from evaporative water loss

increases. The fine interaction between local temperature

and its impact on the frog’s ability to maintain moisture

is a key to amphibian homeostasis (Navas 1996a,b) and

may result in unpredictable outcomes (Burrowes et al.

2020). In fact, temperature was a significant predictor of

coqui house occupancy (Table 2), and the data revealed

that frogs occupied artificial habitats significantly more

often during the cool-dry season (Fig. 7A, shaded box).

Low temperatures and drought are unfavorable condi-

tions for tropical frogs, especially terrestrial direct-devel-

opers that depend on ambient moisture for rehydration

(Duellman and Trueb 1994; Navas 1996b). Studies in

Puerto Rico have shown that both nocturnal activity and

Amphib. Reptile Conserv.

reproduction of E. coqui decrease during the dry season

(reviewed by Joglar 1998) and that dry periods negative-

ly affect their response to pathogens such as the chytrid

fungus (Longo et al. 2010). Thus, we infer that the use of

artificial retreats, such as the bamboo houses provided in

this study, may have contributed to mitigating the effects

of the hurricane.

Bamboo houses by day had a higher occupancy rate

(Fig. 8), suggesting that the need for arboreal understory

habitat that would serve as daytime retreat and nesting

site was of higher demand than the nocturnal perching

sites for which the PVC houses were used (Fig. 9). Thus,

the results show that although coquis can survive hur-

ricane devastation by hiding in the complexity of fallen

vegetation (Woolbright 1991), they will use arboreal

habitat if available. The preference for the more enclosed

and protected bamboo houses during the daytime (when

temperatures are higher, and the risk of dehydration is

greater) 1s expected under the devastated canopy condi-

tions caused by Hurricane Maria. Although PVC houses

were used less often (Table 2), they were used at night

throughout this study, often at an occupancy rate of 10%

(Fig. 8B), suggesting that these open cylinders provided a

sheltered perching site that may also benefit coqui males

by, for example, amplifying calling sound at a time when

other natural microhabitats were unavailable (Fig. 9).

Time since the hurricane, which is a proxy of forest

recuperation, was also a predictor of the occupancy rate

of coqui houses (Table 2), resulting in a significant bell-

shaped response in daytime usage (see Fig. 8A). A rapid

increase in usage at the beginning of the study, despite a

potential learning curve to discover the “new habitats,”

highlights the role of coqui houses in providing arboreal

retreats and nesting sites soon after hurricane damage

(recovery stages 1—2, Table 1). However, the peak in fre-

quency of usage occurs later, in the midst of the cool-dry

season when environmental conditions are most stressful

to the frogs (Fig. 8A, shaded box). The gradual decrease

in usage by the end of the study may be related to for-

est structure recovery since the hurricane. This pattern is

confirmed by the significant interaction of temperature

and time since the hurricane as predictors of the occu-

pancy rate of artificial coqui houses (Table 2). With field

notes and photographs, a chronological succession of the

forest was documented, from over 90% canopy depletion

to the recovery of foliage in standing trees to over 70%,

and then to the replacement of invasive understory veg-

etation by native saplings that contribute more structure

and shade by the end of 2019 (Table 1, Fig. 2). As the for-

est recuperates from the initial severe damage, restored

natural habitat in standing trees is expected to serve as

appropriate calling, perching, retreat, and nesting sites,

and thus, the frogs would have a lesser need to occupy

artificial habitats.

The rate of usage of bamboo houses by coqui frogs

after Hurricane Maria (13-27%) was similar to that

found in other studies that were successful in using artifi-

June 2021 | Volume 15 | Number 1 | e274

Burrowes et al.

cial nesting sites as conservation methods. For example,

Grubb and Bronson (1995) obtained 25% usage with the

Carolina Chickadee birds in Ohio, USA; and in Beni-

dorm, Spain, the European Storm Petrels used 29% of

the artificial nesting sites (De Leon and Minguez 2003).

However, in Puerto Rico, a previous study showed that

coqul frogs used up to 46% of the bamboo houses placed

to enhance habitat in the lowland forests (Stewart and

Pough 1983). With that amount of usage in undamaged

forest (Stewart and Pough 1983), we expected to see a

greater effect of artificial houses on the abundance of co-

quis in the experimental transect versus the control after

Hurricane Maria. However, this was not the case (Fig.

10) probably because retreat sites are not a limiting fac-

tor for coquis after hurricane disturbances (Woolbright

1991). The increase of vegetative debris on the forest

floor may have provided hiding places that were good

alternatives to arboreal sites, allowing the frogs to cope

with the other stressful conditions that the lack of cano-

py imposed. Other methodological caveats of this study

may have contributed to the inability to show an effect

of coqui houses on the number of frogs active at night.

Since quantitative methods were not used to accurately

compare the degree of canopy devastation between tran-

sects immediately after the hurricane, it is possible that

the control transect had better conditions for the coquis

in terms of perching, retreat, and reproduction sites than

the experimental transect. It is also plausible that the

number of coqui houses provided in this study was too

few; while 44 coqui houses were set in 150 m*, Stewart

and Pough (1983) placed 100 of them in 100 m? plots. A

greater number of artificial habitats may be needed, per-

haps as a threshold, before a positive impact on popula-

tion numbers can be observed. Finally, visual encounter

surveys may not be the best way to count frogs after hur-

ricane devastation of the habitat, particularly for a gener-

alist species like E. coqui, because adults and especially

juveniles can hide amongst the fallen debris where they

would remain undetected.

Conclusions and Perspectives

This project aimed to determine whether a tropical forest

frog population affected by an environmental catastro-

phe, such as a category 4—5 hurricane, would use artifi-

cial habitats set to augment arboreal forest structure. The

findings reveal an increase in the forest temperature as a

consequence of canopy blowdown, provide a description

of the initial damage and early recovery stages of the for-

est (Table 1, Fig. 2), and highlight the importance of time

since the hurricane, local climate, and the type of habitat

on the frequency of usage of artificial habitats by coqui

frogs. Although the data did not confirm that this kind of

management intervention would result in an increase in

the number of frogs, they did show that artificial habitats

were used, and that occupancy rate increased significant-

ly during the most stressful environmental conditions,

Amphib. Reptile Conserv.

i.e., early after the hurricane and during the cool and dry

season (Fig. 7A), which highlights their potential as a

mitigation strategy after extreme events. Unfortunately,

we missed the opportunity to study the immediate effects

of Hurricane Maria on coqui frog populations because

safety reasons prevented us from sampling in our tran-

sects for several months. Thus, whenever safe and fea-

sible, we recommend beginning studies on the impact of

environmental disasters as soon as possible in order to

record the full range of their consequences and the re-

sponses of the organisms in question.

Acknowledgements.—Our work was conducted under

permits from The Department of Natural and Environ-

mental Resources of Puerto Rico-PR (DRNA: 2018-IC-

067) and Institutional Animal Care and Use Committee

(IACUC: 3012-05-23-2018). Alberto Lopez kindly pro-

vided the photo of the male E. cogui. We acknowledge

support from NSF grant (IOS-2011281) to PA. Burrow-

es, and we are grateful to the many students from Uni-

versity of Puerto Rico who helped conduct field work,

especially J. Pefia, I. Ramirez, and P. Delgado.

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il i iy

aed LOE a ai

a A : ee

Patricia A. Burrowes is a Professor in the Department of Biology at The University of Puerto Rico in San Juan. She received

her Ph.D. at the University of Kansas under the mentorship of W.E. Duellman, and her early-career research interests included

community ecology, reproductive behavior, and population genetics of tropical amphibians. Given the precipitous decline

of amphibian populations worldwide, Patricia has been dedicating her efforts to the study of the factors involved in this crisis,

particularly the ecology of chytridiomycosis under enzootic conditions and the responses of tropical amphibians to climate warming.

Amphib. Reptile Conserv.

June 2021 | Volume 15 | Number 1 | e274

Coqui frogs and Hurricane Maria

Abner D. Hernandez-Figueroa (left) is a recent graduate from the Environmental Sciences program at the University of Puerto

Rico in San Juan. Abner’s interests and research experiences as an undergraduate focused on the conservation and ecological

restoration of amphibians and coral reefs, which are being challenged by severe environmental changes as a consequence of global

climate change.

Gustavo D. Acevedo-Vélez (right) recently obtained his B.S. in the Environmental Science program at the University of Puerto

Rico in San Juan. His research interests include the behavioral and population ecology of amphibians and aquatic invertebrates.

Currently Gustavo is a field technician for a stream flow reduction study on community composition and dynamics at the Luquillo

LTER (Long-Term Ecological Research) site.

Junangel Aleman-Rios (middle) is an undergraduate student at the University of Puerto Rico in San Juan, where he is currently

completing a double major in Biology and Environmental Sciences. His research interests include the behavioral ecology of

amphibians and wildlife diseases, and Junangel expects to continue his studies in veterinary medicine.

Ana V. Longo is an Assistant Professor in the Department of Biology at the University

of Florida in Gainesville, Florida, USA. At every step of her academic career (from

Master’s degree to Postdoc), Ana has had the unique opportunity to be mentored

and supported by three outstanding women herpetologists: Patricia Burrowes, Kelly

Zamudio, and Karen Lips. Ana’s research program aims to identify and quantify

the ecological factors and evolutionary processes that allow amphibians to interact

with their pathogens, parasites, and symbionts. She actively seeks to broaden the

participation of underrepresented minorities in the fields of ecology and evolution

through collaborations, student mentoring, and training.

Amphib. Reptile Conserv. 70 June 2021 | Volume 15 | Number 1 | e274

Amphibian & Reptile Conservation

15(1) [General Section]: 71-107 (e275).

Official journal website:

amphibian-reptile-conservation.org

Amphibian diversity of a West African biodiversity

hotspot: an assessment and commented checklist of the

batrachofauna of the Ivorian part of the Nimba Mountains

‘Kouassi Philippe Kanga, **N’Goran Germain Kouameé, ‘Parfait Zogbassé, ‘Basseu Aude-Inées

Gongomin, ‘Konan Laurent Agoh, 7Akoua Michele Kouameé, 7Jean Christophe B.Y.N. Konan,

?Abouo Béatrice Adepo-Goureéne, 7Germain Gouréne, and *Mark-Oliver Rodel

'Université Jean Lorougnon Guédé, Laboratoire de Biodiversité et Ecologie Tropicale (BioEcoTrop), Daloa, BP 150, COTE D’IVOIRE *Université

Nangui Abrogoua, Péle de Recherche Péche et Aquaculture, UFR-SGE, 02 BP 801, Abidjan 02, COTE D’IVOIRE ?Museum fiir Naturkunde, Leibniz

Institute for Evolution and Biodiversity Science, Invalidenstr. 43, 10115 Berlin, GERMANY

Abstract.—This article provides the first assessment and commented checklist of the anuran diversity of the

Ivorian part of the Mount Nimba Integrated Nature Reserve (MNINR), West Africa. During a period of 81 days

from 18 June 2018 to 17 May 2019, covering both the rainy and dry seasons, 53 amphibian species were

recorded. Among these species, 30.2% were endemic to either the Upper Guinea forest zone or smaller areas

within that biodiversity hotspot. The amphibian fauna of the Ivorian slope of the MNINR is very similar to those

of the Guinean side of Mounts Nimba and the Guinean Simandou Range. Based on the current IUCN Red List

data, several recorded species are of high conservation concern: the Critically Endangered Nimbaphrynoides

occidentalis; the Endangered Hyperolius nimbae; and the Near Threatened Leptopelis macrotis, Leptopelis

occidentalis, and Odontobatrachus arndti. Of particular interest among the survey records were the poorly

known Ptychadena arnei, P. pujoli, and P. submascareniensis. The records of Ptychadena retropunctata and

Arthroleptis crusculum represent first country records for lvory Coast, while the records of Odontobatrachus

arndti and Phrynobatrachus fraterculus are the second records for the country. In contrast to the Guinean and

Liberian parts of Mounts Nimba, the Ivorian part had never been mined or explored for mining, nor do such

plans currently exist. As a result, the study area still holds intact mountain forests that include rare and unique

habitats with exceptional biodiversity, which need to be preserved for future generations. Consequently,

conservation strategies should minimize bush-fires in mountain grasslands, e.g., to protect the viviparous

toad N. occidentalis. At lower elevations, it is important to encourage local activities concerning reforestation

of the previously forested areas and the conservation of the (sacred) village forests.

Keywords. Anura, conservation, endemics, first record, Ivory Coast, UNESCO World Heritage Site, taxonomy, Upper

Guinea forest

Citation: Kanga KP, Kouamé NG, Zogbassé P, Gongomin BAI, Agoh KL, Kouamé AM, Konan JCBYN, Adepo-Gouréne AB, Gouréne G, Rédel MO.

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).

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: 28 April 2021; Published: 27 June 2021

Introduction 2005; Kozak and Wiens 2010; Blackburn 2008; Juarez-

Ramirez et al. 2016; Portik et al. 2016; Doherty-Bone

Worldwide habitat loss and the consequent decline of |= and GvoZdik 2017; Khatiwada et al. 2019; van der Hoek

terrestrial vertebrates have particularly severe impacts et al. 2019; Bittencourt-Silva et al. 2020). This pattern

on amphibians (e.g., Lips 1999; Raxworthy and also applies to the West African highlands, which host

Nussbaum 2000; Hero and Morrison 2004; Stuart et unique and remarkable amphibian species (Kouameé et al.

al. 2004; Crawford et al. 2010). Mountainous regions 2007; Hillers et al. 2008a; Ofori-Boateng et al. 2018).

are particularly important areas for the conservation The Upper Guinean forest zone of West Africa is among

of amphibians since they provide a large number of — the most important global biodiversity hotspots (Myers

different habitat types with high species richness. In — et al. 2000). Within that ecoregion, the Mounts Nimba

particular, the species inhabiting the higher elevations is situated on the borders between Liberia, Guinea, and

are often range restricted or endemic (Herrmann et al. Ivory Coast, and harbors a particularly large number of

Correspondence. *ngoran_kouame@yahoo.fr

Amphib. Reptile Conserv. 71 June 2021 | Volume 15 | Number 1 | e275

Amphibians of the Nimba Mountains (Ivorian part)

endemic species (Schnell 1952; Angel et al. 1954a,b;

Lamotte and Sanchez-Lamotte 1999; Erard and Brosset

2003; Girard 2003; Ineich 2003; Roy 2003; Monadjem

et al. 2013; Decher et al. 2016; Simmons et al. 2021).

Thus, this mountain range, which abruptly rises from

the surrounding plains up to an elevation of 1,752 m asl,

is considered of “Exceptionally High Priority’ for the

conservation of biodiversity in the Upper Guinea forest

zone (Bakarr et al. 2001).

While the amphibian fauna of Mounts Nimba ranks

among the most species rich and most intensively studied

of the amphibian faunas in West Africa (Rodel et al.

2004), almost all research has been done on the Guinean

and Liberian parts of the mountain range (e.g., Guibé

and Lamotte 1958a,b, 1963; Xavier 1978; Rodel et al.

2010; Sandberger et al. 2010; Sandberger-Loua et al.

2018a; Schafer et al. 2019), and none has ever focused

exclusively on the Ivorian part of the mountains. One

particular research focus was on the viviparous toad

Nimbaphrynoides occidentalis (Angel, 1943), which

is a flagship species for the conservation of the area

(e.g., Lamotte 1959; Lamotte and Sanchez-Lamotte

1999; Hillers et al. 2008a; Xavier 2009; Sandberger

et al. 2010; Sandberger-Loua et al. 2016, 2017,

2018b). However, although much research effort

has been directed to the amphibian fauna of the Nimba

Mountains, as indicated by the accumulation of tens of

thousands of amphibian vouchers at the Muséum National

d’ Histoire Naturelle in Paris, new records and new species

descriptions continue to be published from the Guinean

and Liberian parts of Mounts Nimba (Rodel et al. 2009,

2010; Bare et al. 2015; Sandberger-Loua et al. 201 8a).

Thus, there is a pressing need to investigate the Ivorian

part of the Nimba Mountains as well, and such research

seems particularly urgent as the steadily increasing human

population in the area is demanding access to land. As a

result of armed conflicts in Ivory Coast and neighboring

countries, a massive influx of refugees into the forested

areas of western Ivory Coast is increasingly limiting the few

remaining areas of primary rainforests (Woods 2003). The

northern part of Mounts Nimba, shared between Guinea

and Ivory Coast, received strict protection in 1944. The

Ivorian part was gazzetted as an UNESCO World Heritage

Site in 1982, and declared the Mount Nimba Integrated

Nature Reserve (MNINR) by the national law 2002-102

of February 2002, and the property was integrated into the

public domain of Ivory Coast (Lauginie 2007). This paper

presents the results of recent field surveys with the aim

of providing a better understanding of the batrachofauna

of the Ivorian sector of the Nimba Mountains, and this

information will contribute to the long-term protection of

this unique and biodiverse area.

Materials and Methods

Study area. Amphibian surveys were carried out in the

Mount Nimba Integrated Nature Reserve (MNINR),

Amphib. Reptile Conserv.

situated at the westernmost extension of the mountains in

Ivory Coast (07°25’-07°45’N, 008°20’—008°35’W; Fig.

1). Covering 5,000 ha, the MNINR makes up only a small

portion of the Nimba Mountains, while the largest part

(12,540 ha) is located in Guinea. The highest peak of the

Nimba Mountains is the Richard-Molard with an altitude of

1,752 m asl (Lamotte et al. 2003a,b; Laugenie 2007).

The varied geomorphology and the sub-equatorial

climate, with strong seasonal and altitudinal differences,

result in a variety of different microclimates. Fluctuations

of mean annual temperatures range between 22—27 °C

on the mountain bases to 16—21 °C on the peaks. Daily

temperature fluctuations may span more than 20 °C during

the dry season. Temperatures are lowest during the core

rainy season from August to September, and reach highest

values in March and April. The rainy season extends

over eight to nine months, and is only interrupted by a

short dry season from November/December to February/

March. Annual precipitation varies considerably between

the low and high elevations and is highest in the montane

grasslands, where it may reach up to 3,500 mm. In

the dry season a warm, dry and dusty wind, known as

Harmattan, prevails (Lauginie 2007). Humidity in the

rainy season usualy exceeds 80%, but drops below 30%

during Harmattan periods. The Nimba range is a water

reservoir, and the source of more than 50 streams and

rivers, among which the rivers Cavally, Goue, and Nuon

are of regional importance. During the rainy season, the

montane parts of the mountains are mostly enveloped in

clouds. The slopes are predominantly covered with dense

evergreen forests at lower to mid-elevations, giving

way to patches of moist savannah. Higher elevations

(those above ~1,200 m asl) are dominated by montane

grasslands on iron-ore ground (Lamotte 1998; Lamotte

et al. 2003a,b; Lauginie 2007).

Survey sites. Four distinct habitat types are distinguished

with different amphibian assemblages. Their defintions

have been based on elevation and vegetation. Habitat

A (Fig. 2) comprises montane grasslands present at the

highest elevations. Habitat B (Fig. 3) 1s a mid-elevation

savannah/grassland. An important species in the montane

grasslands and mid-elevation savannah is Loudetia

kagerensis (Poaceae), which grows on_ iron-oxide

quartzite ground. These open habitats are frequently

affected by fires during the dry season. Habitat C (Fig.

4) is dense, broadleaf and evergreen forests stretching

from lower to mid-elevations (422 to 847 m asl). Torrent

streams cross these mountain forests in ravines, and the

water level of mountain streams decreases considerably

during the dry season. Habitat D (Fig. 5) comprises

altered, former forest habitat that is now inhabited

predominantly by non-forest species. This area is close

to Yéalé village, situated at 377 m asl, about 2 km from

the periphery of the MNINR. The village area is bordered

by islands of bamboo forests, partly intact forests,

degraded forests with large clearings, and thick grassy

June 2021 | Volume 15 | Number 1 | e275

Kanga et al.

3° Ww

GUINEA

ic &

LIBERIA

Legend

pe City

@ village

Gm Road

Stream

State boundaries

Montane grasslands

Savannah grasslands

Dense forests

Altered forests

0123 4Km

Weal oe! Ka A

Yeale

le of

Nimba

OAST

sKouan-Houlé |

Danane

Fig. 1. Geographical location of the Mount Nimba Integrated Nature Reserve, within the westernmost extension of Ivory Coast at

the border crossing point with Guinea and Liberia. Four distinct habitats with different amphibian assemblages based on altitudinal

and vegetation types are indicated: Montane grasslands at the highest elevations (A: red star; Fig. 2); Mid-elevation savannah

grasslands (B: purple star; Fig. 3); Dense, broadleaf and evergreen forests from lower to mid-elevations (C: orange star; Fig. 4); and

Altered forests (D: green star; Fig. 5). The inset figure indicates the location of Ivory Coast (green patch) on the African continent.

and shrubby vegetation around houses. Cocoa and coffee

plantations, small-scale subsistence farming, cultivating

plantains, cassava, and corn, as well as swamps used

for rice cultivation, dominate large parts of the village’s

sourroundings.

Field work, sampling effort, and vouchers.

Despite the absence of the panzootic chytrid fungus

Batrachochytrium dendrobatidis (Bd) from West Africa

west of the Dahomey Gap (Penner et al. 2013; Zimkus et

al. 2020), as a precaution new or disinfected equipment

Amphib. Reptile Conserv.

was always used on each survey. Animals were mainly

found opportunistically through visual encounter surveys

(Heyer et al. 1994; Rodel and Ernst 2004), supplemented

by acoustic surveying, lifting logs and rocks, peeling

away bark, scraping through leaf litter, tufts, grasses,

and broad-leaved trees, and searching around or within

water-filled tree holes. Furthermore, potential breeding

sites were checked for tadpoles by dip-netting. Because

amphibians were not marked, repeated observations of

a given individual in multiple visits cannot be excluded.

Surveys of all accessible habitats were conducted by

June 2021 | Volume 15 | Number 1 | e275

Amphibians of the Nimba Mountains (Ivorian part)

Fig. 2. Montane grasslands (habitat A) at 1,241 m asl.

C) at 847 masl.

five people during both day (0830-1230 h GMT) and

night (1 800—2200 h GMT). Field work was conducted

in the rainy and dry seasons, and included a total of 81

days from 18 June 2018 to 17 May 2019. Habitats B, C,

and D were each investigated 56 times (day and night)

during the rainy season, and 25 times (day and night)

in the dry season. The montane grasslands (A) were

investigated 12 times in the rainy season and seven times

in the dry season, both during daytime only (Table 1).

Night searches could not be conducted in this habitat,

as overnight stays were not possible, and decending the

mountain at night was too dangerous.

The overall sampling effort involved 3,240 person-

hours. The geographical coordinates using the WGS84

datum for each site were recorded with a hand-held GPS

Fig. 3. Mid-elevation savannah (habitat B).

Fig. 5. The Yéalé village at the foothills of Mounts Nimba

showing altered former forest habitat (habitat D). The

mountains are visible in the background.

device (Garmin 20 etrex). The observations for each

species are summarized below, and the nomenclature used

herein follows Channing and Rodel (2019). Amphibians

were captured by hand and identified to species level. All

individuals were photographed, measured, sexed, and

if not retained as vouchers, released in their respective

habitats. The symbols in “*” refer to records that probably

comprises several species in the Tables and Appendix.

Snout-urostyle-lengths (SUL) were taken with dial

calipers (accuracy + 0.5 mm). Voucher specimens were

euthanized in a_1,1,1-trichloro-2-methyl-2-propanol

hemihydrate (MS222) solution, preserved in 80% ethanol,

and deposited at the Jean Lorougnon Guédé University,

Daloa, Ivory Coast. The numbers of retained vouchers

(NGK) are listed with the species accounts.

Table 1. Number of daily searches, during the 81 day period from 18 June 2018 to 17 May 2019, in each of the four habitat types (A,

B, C, and D) in Mount Nimba Integrated Nature Reserve. The habitat types are described in the “Survey Sites” section in Materials

and Methods.

Habitat A Habitat B Habitat C Habitat D

ight ight

June 2021 | Volume 15 | Number 1 | e275

ight

rstofulo [sta

Amphib. Reptile Conserv. 74

Kanga et al.

Statistics. As most data were collected opportunistically

(and are thus not strictly quantitative), only the Chao2 and

Jackknifel estimators, both based on presence/absence

data and for all habitats, were used to calculate expected

species richness and thus sampling efficiency, using the

software EstimateS (Colwell 2006). Calculations were

based on the daily species lists (81 days of survey work)

for all 53 amphibian taxa recorded in MNINR. To avoid

order effects, calculations were based on 500 randomized

runs of the daily species lists. The Serensen’s similarity

index (I]), which varies from 0 to 1 (Sorensen 1948;

Wolda 1981), was used to determine species similarity

between the four habitat types (A, B, C, and D).

Likewise, I] was used for pairwise comparisons between

species overlap with the MNINR (this study) and eight

nearby and surrounding sites in the western part of the

Upper Guinean hotspot that were previously surveyed

(Chabanaud 1920, 1921; Parker 1936; Guibé and Lamotte

1958a, 1963; Laurent 1958; Schiotz 1967; Taylor 1968;

Bohme 1994a,b; Lamotte and Ohler 1997, 2000; Rodel

2003; Rodel and Ernst 2003; Rodel and Bangoura 2004;

Rodel et al. 2004; Ernst et al. 2006; Hillers et al. 2008a;

Bare] et al. 2015; Sandberger-Loua et al. 2018; Schafer

et al. 2019). The Hyperolius sp. by Hillers et al. (2008a)

from Fouta Djalon was H. occidentalis (N.G. Kouamé,

pers. obs.). Because the intraspecific morphological

variation of frogs from the West African Arthroleptis

poecilonotus-complex (< 30 mm snout-urostyle length)

overlaps with interspecific variation, it 1s currently not

possible to distinguish these frogs at the species level

based on morphological characteristics alone (Rodel

and Bangoura 2004). Advertisement calls of our records

hint that these squeaker frogs may actually represent

several taxa, but are treated herein as one taxon,

termed Arthroleptis poecilonotus-complex. Apart from

Hyperolius sp., the records of frogs not identified to the

species level were excluded from the calculations of the

Sgrensen index.

Results

Species Richness and Faunal Similarities

A total of 53 anuran species were recorded in MNINR.

A list of all taxa with site records, known general habitat

preferences, distribution range, and the current IUCN

Red List category is provided in Table 2. Concerning

range, our definition of West Africa follows Penner et

al. (2011), 1.¢., a region extending from Senegal in the

west to the Nigerian Cross River in the east. Using the

estimators, we calculated 53 (sd: + 0.62; Chao 2) and

54 (sd: 1.39; Jack-knife 1) anuran species to occur in

the study area. Thus, we found almost the entirety of the

species richness (100% and 96.29%, respectively) for the

Ivorian sector of Mounts Nimba.

Approximately one-third (16 spp., 30.2%) of the

recorded species are restricted to the Upper Guinean

Amphib. Reptile Conserv.

forest zone, while another one-third (16 spp., 30.2%)

are even further limited to the western part of this

biodiversity hotspot. Two species (3.8%), Hyperolius

nimbae and Nimbaphrynoides occidentalis, are endemic

to the Nimba area; while eight species (15.1%) have a

larger West African range. Eleven species (20.75%) are

known to occur beyond West Africa (Table 2).

Over one-quarter of the species encountered (14 sp.,

26.4%) require forest habitats. Among them, eight species

(15.1%) are typical savannah specialists; five (9.4%)

occur in farmbush (degraded forest) habitats, while 11

(20.8%) are known to inhabit savannah and farmbush

habitats; nine (17%) are known from forest and savannah

habitats; and two species (3.8%), Phrynobatrachus

gutturosus and Ptychadena arnei, have been recorded

across the entire broad habitat range from savannah and

farmbush to forest (Table 2).

Concerning the habitat specific species richness

in MNINR, the following species numbers were

recorded in each of the four distinct habitats: four

species in A, eight in B, 21 in C, and 32 in D. Two

species (Phrynobatrachus tokba and _ Ptychadena

submascareniensis) were common to habitats A and B,

while three (Astylosternus occidentalis, Sclerophrys

maculata, and Phrynobatrachus tokba) were common to

B and C. Three species (Leptopelis viridis, Sclerophrys

maculata, and Hoplobatrachus occipitalis) occured in

B and D, and five (Arthroleptis poecilonotus-complex,

Leptopelis macrotis, Sclerophrys maculata, Hyperolius

chlorosteus, and Kassina cochranae) were found in C

and D (Table 2). Habitat A showed the highest species

overlap/similarity (I]-value: 0.33) with habitat B; habitat

B shared a mean species overlap/similarity (I]-value:

0.21) with habitat C, and the lowest species overlap/

similarity (I]-value: 0.21) with habitat D (I]-value: 0.15);

while habitat C shared a low species overlap/similarity

(I]-value: 0.19) with habitat D.

One species, Phrynobatrachus tokba, occurs in forest,

farmbush habitats, and montane grasslands; another

species (Ptychadena submascareniensis) inhabits

savannah and montane grasslands; while two species

(3.8%), Arthroleptis crusculum and Nimbaphrynoides

occidentalis, are confined to montane grasslands only

(Table 2).

With respect to the IUCN status of threatened

species, one species (Nimbaphrynoides occidentalis)

is ranked as Critically Endangered; one species,

Hyperolius nimbae, is l\isted as Endangered; four

(Arthroleptis crusculum, Leptopelis macrotis, L.

occidentalis, and Odontobatrachus arndti) are Near

Threatened; and four (Hyperolius soror, Ptychadena

arnei, P. pujoli, and P. submascareniensis) are Data

Deficient.

The results of the Sorensen’s similarity index for

pairwise comparisons between the MNINR (this study)

and eight nearby and surrounding sites from the western

part of the Upper Guinean hotspot are presented in

June 2021 | Volume 15 | Number 1 | e275

Amphibians of the Nimba Mountains (Ivorian part)

Table 2. Amphibian species recorded in the Mount Nimba Integrated Reserve with respective habitat records, known habitat

preferences, distribution range, and IUCN Red List category. Habitats A, B, C, and D are described in the “Survey Sites” section in

Materials and Methods. AF = Africa (any range beyond West Africa), WA = West Africa (defined as the area west of the Cross River

in Nigeria), UG = Upper Guinea (defined as forest zone west of the Dahomey Gap), wUG = western Upper Guinea (defined as any

range from western Ivory Coast or beyond the western part of this country), E = endemic to Mounts Nimba; S = savannah, MG =

montane grasslands, FB = farmbush (degraded and secondary forest), F = forest; [UCN categories: LC = Least Concern, NT = Near

Threatened, EN = Endangered, CR = Critically Endangered, NE = Not Evaluated by IUCN.

anitesandspesies aT [c] > [ar[ wa luc] wole]s [mo | ro] F | Red iit

Amphib. Reptile Conserv. 76 June 2021 | Volume 15 | Number 1 | e275

Kanga et al.

Table 2 (continued). Amphibian species recorded in the Mount Nimba Integrated Reserve with respective habitat records, known

habitat preferences, distribution range, and IUCN Red List category. Habitats A, B, C, and D are described in the “Survey Sites”

section in Materials and Methods. AF = Africa (any range beyond West Africa), WA = West Africa (defined as the area west of the

Cross River in Nigeria), UG = Upper Guinea (defined as forest zone west of the Dahomey Gap), wUG = western Upper Guinea

(defined as any range from western Ivory Coast or beyond the western part of this country), E = endemic to Mounts Nimba; S =

savannah, MG = montane grasslands, FB = farmbush (degraded and secondary forest), F = forest; IUCN categories: LC = Least

Concern, NT = Near Threatened, EN = Endangered, CR = Critically Endangered, NE = Not Evaluated by IUCN.

Pomniomnrveses Te Pea [are] we [eee EPS [me [| Be

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Phrynobatrachus

natalensis

Phrynobatrachus

phyllophilus Me ie

Phrynobatrachus tokba

Pipidae

f a

Xenopus tropicalis

Ptychadenidae

Ptychadena arnei

Ptychadena bibroni

Ptychadena longirostris

Ptychadena oxyrhynchus

Ptychadena pujoli

Ptychadena pumilio

Ptychadena retropunctata

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Table 3. Among the Ivorian sites, the Tai National Park

had the highest similarity (I]-value: 0.62) with MNINR

concerning amphibian assemblage composition. More

than half of the anuran fauna of MNINR was also shared

with the Mount Sangbé National Park (I]-value: 0.58)

and the Mount Péko National Park (I]-value: 0.54), two

mountainous areas in western Ivory Coast. Naturally,

Amphib. Reptile Conserv.

our survey area was most similar to the amphibian fauna

of the Guinean part of Mounts Nimba (79% similarity).

With 72% and 67% similartity, the Guinean Simandou

range and Diécké Classified Forest likewise had faunas

that were very similar to the MNINR. In contrast the

Fouta Dyjalon, the westernmost part of the Upper Guinea

highlands, comprising savannah, limited forest, and

June 2021 | Volume 15 | Number 1 | e275

Amphibians of the Nimba Mountains (Ivorian part)

Table 3. Sorensen’s Similarity Value (I]) between the anuran fauna of the Mount Nimba Integrated Reserve and nearby and sur-

rounding sites from the western part of the Upper Guinean hotspot of West Africa, including respective species richness. Area ab-

breviations: DCF = Diécké Classified Forest; FD = Fouta Djalon Highlands; MB = Mount Béro; MN = Mount Nimba; MP = Mount

Péko National Park; MS = Mount Sangbé National Park; SR = Simandou range; ZCF = Ziama Classified Forest. Literature records

were adjusted to reflect recent taxonomic changes.

Number of

Area Species richness species common

with this study

MP (Ivory Coast) 29 22

MS (Ivory Coast) 44 28

TNP (Ivory Coast) 53 33

DCF (Guinea) 39 31

FD (Guinea) 26 15

MB (Guinea) 28 D2

MN (Guinea) 56 43

SR (Guinea) 52 38

ZCF (Guinea) 31 20

rivers with waterfalls, only shares 38% of the species

with MNINR (Table 3).

Species Accounts

After each species name the numbers of retained vouchers

(NGK-Nimba) are listed.

Arthroleptidae

Arthroleptis crusculum Angel, 1950

Evening Squeaker

Material: Two males, NGK-Nimba 0019, NGK-

Nimba 0130 (Fig. 6A). Comments: On a rainy day

they were found together with a juvenile Nimba Toad

(N. occidentlais) below a stone in montane grassland

(07°35.555’N, 008°25.788’ W; 1,235 m asl). These males

had an oval to slender elongated body and measured 15.5

and 21.0 mm SUL, respectively. Arthroleptis crusculum

always possesses a granular to warty dorsal skin. Its habitat

differs from other species of the genus which are present

on Mounts Nimba. Arthrolpetis nimbaensis, A. langeri,

and A. krokosua, and occur in rainforest and/or farmbush

(Guibé and Lamotte 1958b; Rédel et al. 2009; Adum et al.

2011; Nopper et al. 2012; Sandberger-Loua et al. 201 8a).

Arthrolpetis crusculum occurs in high elevation grasslands

up to 1,750 m asl during the rainy season and seems to

survive the dry season in gallery forests and at the edges of

marshes (Guibé and Lamotte 1958b).

Arthroleptis poecilonotus-complex

Mottled Squeaker

Material: Two males, NGK-Nimba 0021 (Fig. 6B),

Amphib. Reptile Conserv.

T]-value

(Sorensen) pounce

0.54 Rodel and Ernst (2003)

0.58 Rodel (2003); Barej et al. (2015)

0.62 Rodel and Ernst (2003); Ernst et al. (2006)

0.67 Rodel et al. (2004)

0.38 Hillers et al. (2008a); Barej et al. (2015)

0.54 Rodel et al. (2004)

Guibé and Lamotte (1958a, 1963); Laurent

(1958); Schiotz (1967); Lamotte and Ohler (1997,

0.79 2000); Rodel et al. (2004); Barej et al. (2015);

Sandberger-Loua et al. (2018); Schafer et al.

(2019)

0.72 Parker (1936); Taylor (1968); Rodel and

Bangoura (2004)

0.48 Chabanaud (1920, 1921); Bohme (1994a,b)

NGK-Nimba 0022. Comments: These squeaker frogs

were widespread in the forest area as well as in farmlands

within the vegetation, where several concealed males

emitted their insect-like chirping calls which cannot be

assigned to any morphotaxa. They occured from 425 to

847 m asl. A male of 23.0 mm in SUL was captured in

a patch of forest and retained as voucher (07°35.233’N,

008°25.190’W; 847 m asl). Another male voucher

(SUL 24.0 mm) was recorded in an agricultural area

(07°31.928’N, 008°25.401’W; 425 m asl). Arthroleptis

poecilonotus-complex are larger than A. langeri but

smaller than A. krokosua. The two vouchers may comprise

different species, of which one could be conspecific

with A. nimbaensis. However, currently that cannot be

clarified (see Rodel and Bangoura 2004; Channing and

Rodel 2019). All of these Arthroleptis species, apart from

A. crusculum, may have similar habitat requirements

(Rodel et al. 2009; Adum et al. 2011; Nopper et al. 2012;

Sandberger-Loua et al. 201 8a).

Astylosternus occidentalis Parker, 1931

Western Night Frog

Material: Two males, NGK-Nimba 0014, NGK-Nimba

0023, and one female, NGK-Nimba 0024 (Fig. 6C).

Comments: While A. occidentalis was _ previously

mostly recorded in patches of lowland forests (Rodel

and Branch 2002; Rodel and Bangoura 2002; Ernst

and Rodel 2006; Hillers and Rédel 2007; Hillers et al.

2008b; Rodel and Glos 2019), on Mounts Nimba the

species occurs in altitudinal forest habitats as well (Guibé

and Lamotte 1958a). During the night several active

individuals were detected among leaf litter in a patch of

forest (07°35.233’N, 008°25.190’W; 847 m asl), close to

June 2021 | Volume 15 | Number 1 | e275

Kanga et al.

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complex female (B); Astylosternus occidentalis female (C),; Cardioglossa occidentalis male (D); Leptopelis macrotis male (FE);

Leptopelis occidentalis male (F); Leptopelis spiritusnoctis female (G);, Leptopelis viridis female (H).

Amphib. Reptile Conserv. 79 June 2021 | Volume 15 | Number 1 | e275

Amphibians of the Nimba Mountains (Ivorian part)

fast-flowing streams. Male SULs ranged from 45.0-50.2

mm (N = 3), while females measured from 46.0-61.0

mm (N = 9). The majority of specimens had a dark brown

dorsum, although one female exhibited an orange color.

In contrast to the general forest habitat requirements of

this species we found some, presumably migrating, frogs

at night, in the core rainy season near a crystal-clear

stream in predominantly grassy savannah (07°35.453’N,

008°24.957’W; 843 m asl). During the day, they were

hidden underneath stones.

Cardioglossa occidentalis Blackburn, Kosuch,

Schmitz, Burger, Wagner, Gonwouo, Hillers, and

Rédel, 2008

Western Long-fingered Frog

Material: Three males, NGK-Nimba 0025, NGK-

Nimba 0026 (Fig. 6D), NGK-Nimba 0027. Comments:

Cardioglossa occidentalis is a nocturnal leaf litter frog,

distributed along forest streams from Sierra Leone

to Ghana (Rodel et al. 2001; Rodel and Branch 2002:

Ernst and Rodel 2006; Blackburn et al. 2008; Hillers et

al. 2008c). During the night, males frequently emitted

insect-like calls (see Rodel et al. 2001), and were well

concealed below leaf litter along forest streams. Three

males were collected near a small stream running through

a slightly degraded forest patch that was dominated by

bamboo (07°32.993’N, 008°24.753’ W; 425 m asl). Their

SULs ranged from 27.0—29.0 mm.

Leptopelis macrotis Schietz, 1967

Large-eared Tree Frog

Material: Three males, NGK-Nimba 0017, NGK-

Nimba 0018, NGK-Nimba 0131 (Fig. 6E). Comments:

Leptopelis macrotis 1s one of the largest species in the

genus. It occurs in primary forests, preferentially at the

edges of streams, from eastern Sierra Leone to Ghana

(Schiotz 1967; Rédel et al. 2014; Channing and Rodel

2019). In Ivory Coast, as in its entire range, the species

is threatened due to forest degradation and conversion,

e.g., two of its Ivorian sites (see R6del and Branch 2002)

have been recently converted into rubber plantations

(P.J. Adeba, pers. comm.). During this survey, only three

males of L. macrotis were recorded, two of which were

found during the dry season. Both frogs (45.5 and 48.5

mm SUL) were perched on a branch of a broad leaf, at

~2.5 m height, close to a large stream (07°33.121’N,

008°25.036’W; 422 m asl). The third male (42.5 mm

SUL), in contrast, was found in a degraded forest during

the rainy season. This male was perched on a branch,

at 75 cm above the ground, close to a large stream

(07°31.932’N, 008°25.508’ W; 387 m asl).

Leptopelis occidentalis Schietz, 1967

Western Tree Frog

Material: Male, NGK-Nimba 0016 (Fig. 6F).

Comments: Leptopelis occidentalis is primary a

rainforest treefrog, preferring forests near streams, and

Amphib. Reptile Conserv.

ranging from western Ghana, through Ivory Coast to

Liberia (Schigtz 1967; Rodel et al. 2005; Hillers and

Rodel 2007; Hillers et al. 2009; Channing and Rodel

2019). After sunset (1830 h GMT), a male (41.5 mm

SUL) with a uniform green dorsum was found perching

on a shrub at the edge of a forest clearing along a stream

(07°33.121°N, 008°25.036’ W; 422 m asl). The species is

known from several forests in Ivory Coast (Schietz 1967;

Rodel and Branch 2002; Ernst and Rodel 2008).

Leptopelis spiritusnoctis Rédel, 2007

Ghostly Tree Frog

Material: Female, NGK-Nimba 0087 (Fig. 6G).

Comments: Leptopelis spiritusnoctis inhabits patches

of degraded and primary forests, from Sierra Leone to

Nigeria (Schiotz 1967; Rodel 2007; Rodel et al. 2014).

During this survey we collected only one female (33.5

mm SUL) in a tree at the edge of a stream (07°33.121°N,

008°25.036’W; 422 m asl).

Leptopelis viridis (Giinther, 1869)

Green Tree Frog

Material: One male, NGK-Nimba 0083, and one female,

NGK-Nimba 0086 (Fig. 6H). Comments: Leptopelis

viridis is a savannah frog, which is also encountered in

herbaceous vegetation from the semi-decideous forest

zone. It ranges across the northern part of sub-Saharan

Africa (Schiotz 1967; Rédel 2000; Channing and Rodel

2019). The species was recorded within grassland at the

foot of Mounts Nimba (07°35.453’N, 008°24.957’W;

843 m asl). Additional populations were recorded in the

Yéalé village, on shrubs, palm trees, and herbaceous

plants (07°31.928’N, 008°25.401’W; 425 m asl). The

males measured 32.0—34.0 mm (N = 6), while a single

recorded female reached 36.0 mm SUL.

Bufonidae

Nimbaphrynoides occidentalis (Angel, 1943)

Nimba Toad

Material: No voucher. Comments: Nimbaphrynoides

occidentalis 1s a unique toad, being viviparous and

endemic to a very limited range of a total of 4 km? on the

ridges of the Mounts Nimba between Liberia, Guinea,

and Ivory Coast (Lamotte 1959; Lamotte and Sanchez-

Lamotte 1999; Hillers et al. 2008b; Sandberger-Loua et

al. 2016, 2017). The toads live in montane grasslands

above 1,200 m asl, where they go into dormancy during

the dry season (Lamotte 1959; Hillers et al. 2008:

Sandberger et al. 2010). Nimbaphrynoides occidentalis

comprises two subspecies isolated by a forested mountain

ridge: the larger N. occidentalis liberiensis 1s restricted

to one site in Liberia, while N. occidentalis occidentalis

occurs in a few sub-populations in Guinea and Ivory

Coast (Sandberger et al. 2010). We found N. occidentalis

occidentalis at 1,235 m asl, on very humid steep slopes

in the montane grassland (07°35.555’N, 008°25.788’ W;

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Kanga et al.

1,235 m asl). Four juveniles and one female were

discovered under rocks. The sizes of the juveniles ranged

from 12.0—14.0 mm while the female measured 23.0 mm.

The basic dorsal color of the juveniles was dark brown,

with a somewhat irregular mixture of light brown and

white spots. Their snout, eyelids, and legs were colored

light brown. The juveniles showed a pattern typical for

adult males (Fig. 7A), whereas the female had a nearly

uniform light brown dorsal color (Fig. 7B). The main

threat for this species in Ivory Coast is bush fires in the

dry season. A detailed assessment of the distribution and

population sizes of N. occidentalis occidentalis from

Ivory Coast is urgently needed.

Sclerophrys maculata (Hallowell, 1854)

Northern Flat-backed Toad

Material: Two males, NGK-Nimba 0043, NGK-

Nimba 0051 (Fig. 7C), and one female, NGK-Nimba

0058 (Fig. 7D). Comments: Sclerophrys maculata is a

common toad with flat and granular parotid glands living

in the savannah zone and edges of heavily degraded

forests (R6del 2000; Poynton et al. 2016). Toads were

found in grassy pastures at the foot of Mounts Nimba

(07°35.258’N, 008°25.052’W; 821 m asl), and some

females were also observed occasionally together with S.

togoensis on forest trails (07°32.993’N, 008°24.753’ W;

425 m asl). Further records were obtained in the Yéalé

village (07°31.928’N, 008°25.401’W; 425 m_ asl),

mostly along dirt roads in puddles, around houses, or in

plantations at swamp edges. The body sizes of two males

were 46.0 and 49.0 mm, while females measured 41.5—

69.0 mm (N = 5). During the reproductive period, some

males exhibited a remarkable yellow color.

Sclerophrys regularis (Reuss, 1833)

Common Toad

Material: Female, NGK-Nimba 0031 (Fig. 7E).

Comments: Sclerophrys regularis has prominent,

roundish, and smooth parotid glands. It inhabits a broad

range of habitats from moist and dry savannahs to forest

margins throughout tropical Africa, most often found

around human settlements (R6del 2000; Channing and

Howell 2006). The species was recorded in the rainy

season at Yéalé village (07°31.928’N, 008°25.401’°W;

425 m asl), where some males called in garbage pits

(e.g., Fig. 7F). A female toad measured 118.0 mm, while

a male’s body size was 102.0 mm.

Sclerophrys togoensis (Ahl, 1924)

Togo Toad

Material: Three males, NGK-Nimba 0004, NGK-Nimba

0028, NGK-Nimba 0029 (Fig. 7G), and one female,

NGK-Nimba 0050 (Fig. 7H). Comments: This toad

has a patchy distribution in primary forests from Togo

to Sierra Leone, and mainly breeds in shallow forest

streams during the dry season (Rédel and Bangoura

2004; Rodel et al. 2004). Four specimens were found

Amphib. Reptile Conserv.

in different forest patches. The variable color pattern of

this species has been deescribed by Rodel and Bangoura

(2004), Channing and Rodel (2019), and Gongomin et

al. (2019). Diagnostic is the parallel, straight, narrow,

and angular parotid glands, running parallel to the side

of the body. Two males were encountered in a patch of

dense forest crossed by a shallow stream (07°33.440°N,

008°24.657’ W; 439 m asl). A female was found together

with another male among humid litter on a forest trail

(07°32.993’N, 008°24.753’W; 425 m asl). While the

female measured 64.0 mm, the SULs of the three males

ranged from 40.5—44.5 mm. In Ivory Coast, S. togoensis

is highly threatened by deforestation (Gongomin et al.

2019).

Conrauidae

Conraua alleni (Barbour and Loveridge, 1927)

Allen’s Giant Frog

Material: Two unsexed, NGK-Nimba 0057 (Fig. 8A),

NGK-Nimba 0058, and two females, NGK-Nimba 0059

(Fig. 8B), NGK-Nimba 0073. Comments: Conraua

alleni is a highly aquatic frog, which inhabits slow- to

fast-flowing forest streams, from lowlands to montane

forest areas. Records are known from eastern Guinea and

Sierra Leone, through Liberia to western Ivory Coast,

with an isolated population occurring 1n western Ghana

(Barbour and Loveridge 1927; Guibé and Lamotte

1958a; Lamotte and Perret 1968; R6del 2003; Rodel and

Bangoura 2004; Channing and Rodel 2019; Rodel and

Glos 2019; Schafer et al. 2019). Some of these populations

may comprise cryptic taxa (see R6del and Branch 2002;

Hillers et al. 2008a). We found C. alleni populations

in streams intersecting forest patches (07°35.258’N,

008°25.052’W; 821 m asl). Other individuals were

heard calling in a very impressive torrent stream in mid-

elevation forest (07°34.652’N, 008°24.966’W: 716 m

asl). The bird-like whistles were heard during day and

night, with peaks after sunset (around 1841 h GMT).

Additional populations were found at night in pools of

a slow running stream with a sandy and rocky bottom.

This stream crossed a slightly degraded forest patch

dominated by bamboo (07°32.993’N, 008°24.753’W;

425 m asl), where a total of 14 adult frogs were caught.

Through palpation of the lower abdomen, two of them

were identified as gravid females. Their body size was

52.8-54.0 mm. The remaining 12 frogs ranged from

51.1-—55.6 mm but could not be sexed. One adult, kept in

captivity for two months, preyed on locusts, ants, spiders,

caterpillars, and butterflies that were floating on the

water surface. All adult C. a//eni had a clear interorbital

line, however, their back pattern varied from a darker

brown with black dots and reddish legs to frogs with

orange patches on the darker brown ground. In contrast

to other described C. alleni (e.g., Channing and Rodel

2019), the venter of our frogs was golden yellow or beige

to pinkish, and the thighs had a pink ventral color. The

June 2021 | Volume 15 | Number 1 | e275

Amphibians of the Nimba Mountains (Ivorian part)

Fig. 7. Bufonids from Mount Nimba Integrated Nature Reserve: Nimbaphrynoides occidentalis juvenile (A); N. occidentalis female

(B); Sclerophrys maculata male (C), the yellow color is only exhibited by some males during breeding; S. maculata female (D),; S.

regularis male (E); S. regularis female (F); S. togoensis male (G); S. togoensis female (H).

Amphib. Reptile Conserv. 82 June 2021 | Volume 15 | Number 1 | e275

Kanga et al.

| a = - . ’ ‘ . , . : ee A, eta ~

| —s ¥ a Sal 7 ' : apes ee

Fig. 8. Conrauid, dicroglossid, hemisotid, and hyperoliids from Mount Nimba Integrated Nature Reserve: Conraua alleni (A-B);

Hoplobatrachus occipitalis (C); Hemisus marmoratus (D); Afrixalus dorsalis (E), A. fulvovittatus (F); Hyperolius chlorosteus

(G-H).

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Amphibians of the Nimba Mountains (Ivorian part)

throat was dark, and pinkish with reddish-brown dots or

uniform beige. The taxonomic status of these frogs and

the populations from nearby Mount Sangbé National

Park (Rédel 2003) should be investigated.

Dicroglossidae

Hoplobatrachus occipitalis (Giimnther, 1858)

African Tiger Frog

Material: Female, NGK-Nimba 0005 (Fig. 8C).

Comments: Hoplobatrachus occipitalis is a_ large

aquatic frog that is widely distributed in savannahs and

disturbed forests across tropical Africa (Channing and

Rodel 2019). A female was caught at night near a stream

within pastures (07°35.453’N, 008°24.9577'W; 843 m

asl). Males were heard calling at night, concealed in rice

paddies in Yéalé village (07°31.928’N, 008°25.401°W;

425 m asl). In the rainy season, adult frogs were

occasionally collected and eaten by the local populations

around the MNINR (Zogbassé et al., unpub. data).

Hemisotidae

Hemisus marmoratus (Peters, 1854)

Marbled Piglet Frog

Material: Two males, NGK-Nimba 0085, NGK-Nimba

0089 (Fig. 8D). Comments: Hemisus marmoratus is a

fossorial frog, very common in the savannah ecoystems

of sub-Saharan Africa (Rodel 2000; Channing and Rodel

2019). We found it in the Yéalé village (07°31.928’N,

008°25.401’°W; 425 m asl), among short grasses at

puddles, in leaf litter under cocoa and coffee trees, and

around a manual hydraulic water pump. The body size of

males ranged from 29.0—34.5 mm (N = 4), while females

measured between 33.0—50.5 mm (N = 8).

Hyperoliidae

Afrixalus dorsalis (Peters, 1875)

Striped Spiny Reed Frog

Material: Male, NGK-Nimba 0051 (Fig. 8B).

Comments: Afrixalus dorsalis is a nocturnal leaf-folding

frog, which inhabits a wide range of western African

habitats, such as savannah, farmbush, and swampy areas

at forest edges (Schigtz 1967; Rédel 2000; Channing

and Rodel 2019). We heard many calling males in rice

paddies and in grassy vegetation of swamps, in Yéalé

village (07°31.928’N, 008°25.401’W; 425 masl). Amale

measured 22.0 mm SUL.

Afrixalus fulvovittatus (Cope, 1861)

Banded Spiny Reed Frog

Material: Two females, NGK-Nimba 0066, NGK-Nimba

0067 (Fig. 8F). Comments: Afrixalus fulvovittatus can

be easily recognized by its characteristic reddish-brown

dorsal surface with three light longitudinal stripes joining

on the tip of the snout. The delicate reddish-brown line

Amphib. Reptile Conserv.

in the middle of each light stripe distinguishes it from

the similar looking A. vittiger (Pickersgill 2007). This

nocturnal species prefers the heavily degraded habitats of

the forest zone (Schigtz 1967; Rddel and Glos 2019). In

the Yéalé village, the species was found in rice paddies

and grassy swamps (07°31.928’N, 008°25.401’W; 425 m

asl). The SULs of two females were 19.8—21.0 mm, thus

they were below the described size for A. fulvovittatus

(Schiovtz 1967).

HAyperolius chlorosteus (Boulenger, 1915)

Large Green Reed Frog

Material: Three males, NGK-Nimba 0075 (Fig. 8G),

NGK-Nimba 0076 (Fig. 8H), NGK-Nimba 0088.

Comments: Hyperolius chlorosteus is common along

streams in pristine forest from western Ivory Coast to

Sierra Leone (Schigtz 1967; Channing and Rédel 2019).

In Ivory Coast, it was reported from the rainforest zone,

i.e., the Tai National Park (e.g., Schiotz 1967; Rodel et al.

2002), as well as from the edge of the forest zone in the

Mount Sangbé National Park (Rodel 2003). Records from

Mount Péko National Park (R6del and Ernst 2003) and

the Haute Dodo and Cavally forests (R6del and Branch

2002) may no longer exist. Thus, our records of H.

chlorosteus from Mounts Nimba, confirming the records

by Schigtz (1967), show that the species prevailed, at

least here, through the past 50 years. The species was

frequently recorded at night along streams in the primary

forest. After a heavy rainfall, a vast number of males (N

> 50) were heard calling from high up in tall trees along a

torrent stream (07°34.652’N, 008°24.966’ W; 716 m asl).

Five males were captured, and their body sizes ranged

from 33.2-37.0 mm. They showed some variation of

their back coloration, however, within the range known

for the species (compare Schiotz 1967; Channing and

Rodel 2019). We found one male in sympatry with L.

macrotis in a degraded forest during the rainy season

(07°31.932’N, 008°25.508’W; 387 m asl), perched up at

approximately 1.80 m above the ground, close to a large

stream. Most males, however, called from much higher

Sites.

HAyperolius concolor (Hallowell, 1844)

Uniform Reed Frog

Material: One female, NGK-Nimba 0011 (Fig. 9A), and

two males, NGK-Nimba 0012 (Fig. 9B), NGK-Nimba

0020. Comments: Hyperolius concolor is one of the

most common West African frogs, widespread in a range

of habitats from savannahs and farmbush to degraded

and gallery forests (Schiotz 1967; Rédel 2000). It even

has been reported from urban sites (Kouamé et al. 2015).

Calling males were abundant in rice paddies and grass-

covered edges of ponds (07°31.928’N, 008°25.401’W;

425 m asl). The species is dichromatic (Portik et al.

2019), with females exhibiting a uniform light yellowish-

green back with reddish toe discs (SUL: 32.0-33.0 mm,

N = 2); males in contrast are brownish, often with some

June 2021 | Volume 15 | Number 1 | e275

Kanga et al.

Fig. 9. Hyperoliid frogs from Mount Nimba Integrated Nature Reserve: Hyperolius concolor female (A); H. concolor male (B); H.

fusciventris fusciventris male (C); H. guttulatus female (D); H. lamottei female (E); H. /amottei male (F); H. nimbae female (G);

H. nimbae male (H).

Amphib. Reptile Conserv. June 2021 | Volume 15 | Number 1 | e275

Amphibians of the Nimba Mountains (Ivorian part)

dark patterns between the eyes and on the back (SUL:

23.5—25.5 mm, N = 5).

HAyperolius fusciventris fusciventris Peters, 1876

Dark-bellied Reed Frog

Material: Male, NGK-Nimba 0077 (Fig. 9C).

Comments: Hyperolius fusciventris fusciventris occurs

from western Ivory Coast into neighboring Liberia

and Guinea in farmbush, heavily degraded forests, and

occasionally in gallery forests within humid savannah

(Schiotz 1967). We encountered the species in the Yéalé

village (07°31.928’N, 008°25.401’W; 425 m asl), in an

open but densely vegetated area close to swamps. The

dorsal color varied from uniform brownish over light

green to dense green. Some individuals with greyish-blue

eyes exhibited a uniform yellow dorsal color. The belly

of females was either dark or light grey. All males had a

lighter belly. The male SULs ranged from 23.5—25.0 (N

= 7), females reached 26.5—27.0 mm (N = 3).

Ayperolius guttulatus Giinther, 1858

Spotted Reed Frog

Material: Female, NGK-Nimba 0052 (Fig. 9D).

Comments: Hyperolius guttulatus lives in the humid

savannah zone and clearings within rainforests (Schiotz

1967; Rodel 2000; Assemian et al. 2006; Kouamé et

al. 2015). The species usually reproduces in larger,

permanent ponds with floating vegetation (Schiotz

1967; Kouamé et al. 2015). We found five males calling

at night between rice paddies around the Yéalé village

(07°31.928’N, 008°25.401°W; 425 m asl). A female

measured 37.0 mm.

HAyperolius lamottei Laurent, 1958

Lamotte’s Reed Frog

Material: One female, NGK-Nimba 0074 (Fig. 9E), and

two males, NGK-Nimba 0081 (Fig. 9F), NGK-Nimba

0082. Comments: Hyperolius lamottei is a savannah

frog with a patchy distribution, preferring low grass

habitats, often in higher altitude, e.g., granite inselbergs

in central-southern Ivory Coast, Liberia, Guinea, Sierra

Leone to Senegal (Arnoult and Lamotte 1958; Lamotte

1969, 1971; Schiotz 1967; Rédel et al. 2004). Recently, it

was reported for the first time from Burkina Faso (Ayoro

et al. 2020). In Ivory Coast, the species was recorded

from the Lamto Faunal Reserve (Schietz 1967) and

the Mount Péko National Park (Rodel and Ernst 2003).

However, more recently Adeba et al. (2010) failed to

re-detect the species in Lamto. We found H. /amottei in

large numbers in grassy montane pasture (07°35.258’N,

008°25.052’W; 821 m asl). The species was only active

during the rainy season, when males and females became

active after sunset. They spent the night perching on high

grasses which covered a flooded iron-oxide quartzite

ground. During the daytime all frogs hid within the dense

herbaceous vegetation. Body size of males ranged from

17.5-19.0 mm (N = 4), females measured 21.5—23.5 mm

Amphib. Reptile Conserv.

(N = 3). Color pattern was variable, but within the range

described by Schiotz (1967).

HAyperolius nimbae Laurent, 1958

Nimba Reed Frog

Material: One female, NGK-Nimba 0070 (Fig. 9G),

and two males, NGK-Nimba 0078, NGK-Nimba 0079

(Fig. 9H). Comments: Hyperolius nimbae is a farmbush

frog, endemic to the eastern foothills of Mounts Nimba

(Schiotz 1967). After 47 years, this species was only

recently rediscovered by Kouamé et al. (2016), reporting

small populations from four villages (Dagbonpleu,

Danipleu, Kouan-Houlé, and Zéalé) within the formerly

known range. An even more recent re-investigation in

these villages failed to confirm the species, and its known

habitats had been destroyed due to road expansions,

development, and urbanization (Gongomin et al., unpub.

data). However, during our study, we encountered a

large number of H. nimbae males in the Yéalé village

(07°31.928’N, 008°25.401’W; 425 m asl). The habitat

was within a plantation of cocoa and coffee that edged a

large and deep pond, which exceeded 100 x 70 m. There,

we observed 15 males at night between broad leaved,

evergreen trees of cocoa and coffee, while only one

female was seen perched between the leaves of a palm

tree. The males ranged from 30.0—35.5 mm (N = 15), the

female measured 34.1 mm. Males varied considerably

in color and also showed some differences in pattern

compared to the female. However, both sexes matched

earlier descriptions by Schigtz (1967).

HAyperolius picturatus Peters, 1875

Painted Reed Frog

Material: One female, NGK-Nimba 0104 (Fig. 10A),

and one male, NGK-Nimba 0105 (Fig. 10B). Comments:

Hyperolius picturatus inhabits farmland and forest areas

in various state of degradation, from central Ghana to

Sierra Leone (Schigtz 1967; Rédel 2000; Channing and

Rodel 2019). We found the species in an agricultural

area of the Yéalé village. The basic dorsal color of males

(28.0-31.5 mm, N = 4) was brownish with two light,

broad dorsolateral stripes, while the venter, including

gular glands, was entirely yellow. A female (35.5 mm)

had yellow, broad dorsolateral stripes with some small

yellow spots on its black chin. Its venter likewise was

bright yellow. This widespread taxon shows some color

variation across its range and may comprise two species

(Schiotz 1967; Rodel and Branch 2002; Rodel and Glos

2019).

Hyperolius soror (Chabanaud, 1921)

Soror Reed Frog

Material: One female, NGK-Nimba 0125 (Fig. 10C),

two males, NGK-Nimba 0126 (Fig. 10D), NGK-Nimba

0127. Comments: We heard over a dozen H. soror males

calling during the rainy season in a dense grassy swamp,

edging forest (07°32.993’N, 008°24.753’ W; 425 m asl).

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Kanga et al.

Fig. 10. Hyperoliid frogs from Mount Nimba Integrated Nature Reserve: Hyperolius picturatus female (A); H. picturatus male (B);

H. soror female (C); H. soror male (D); H. cf. sylvaticus male (E); H. cf. sylvaticus male in ventral view (F); H. sp. male (NGK-

Nimba 0068) (G); H. sp. male (NGK-Nimba 0068) in ventral view (H).

Amphib. Reptile Conserv. June 2021 | Volume 15 | Number 1 | e275

Amphibians of the Nimba Mountains (Ivorian part)

However, only one female and two males were caught

there. The species is sexually dimorphic. The female

(25.1 mm SUL) had a light green back, with diffuse,

minute, reddish-brown spots, red lateral markings, a

red stripe from snout to tip of eye, bluish-grey iris,

and toes and fingers, including webbing, were faintly

red with green tips. Its ventral surface was transparent

bluish green. In contrast, both males (SUL 19.1 and 21.1

mm) had a golden iris, a dark red canthal stripe, green

dorsum with diffuse, minute, reddish-brown spots, light

dorsolateral stripes, and green toes and fingers.

Hyperolius cf. sylvaticus Schietz, 1967

Forest Reed Frog

Material: Male, NGK-Nimba 0108 (Fig. 10E-—F).

Comments: Hyperolius cf. sylvaticus was recorded

in an open area in the Yéalé village (07°31.928’N,

008°25.401’W; 425 m asl). However, H. sylvaticus is

known to be a forest species (Schiotz 1967; Rodel and

Branch 2002; Ernst and Rodel 2008), thus it might just as

well be a member of the Hyperolius picturatus-complex.

Hyperolius sp.

Material: Male, NGK-Nimba 0068 (Fig. 10G—H).

Comments: Two Hyperolius sp. were recorded in the

Yéalé village (07°31.928’N, 008°25.401’W; 425 m asl),

in a swampy area with a shallow creek, dense herbaceous

formations, and few shrubs and trees. A male (26.4 mm)

possessed a uniform greyish-brown dorsal color; the

throat was dull blue; the anterior part of the belly was

white, the posterior part greenish to blueish. The ventral

surfaces of limbs were greenish; the discs of toes and

fingers were reddish. The gular gland was indistinct

and smooth. So far, we cannot assign these frogs to any

West African Hyperolius species (e.g., Schiotz 1967;

Channing and Rodel 2019). Molecular and acoustic data

are required in order to resolve the taxonomic status of

this reedfrog.

Kassina cochranae (Loveridge, 1941)

Cochran’s Running Frog

Material: Four males, NGK-Nimba 0139, NGK-Nimba

0140, NGK-Nimba 0141 (Fig. 11A), NGK-Nimba 0142.

Comments: Kassina cochranae is an arboreal forest and

farmbush dweller, ranging from the rainforest edge into

the moist savannah zone from western Ivory Coast to

eastern Sierra Leone (Schigtz 1967; Rédel et al. 2002).

During the rainy season, we heard a vast number of

males calling concealed in dense vegetation, close to a

grassy swamp (habitat C: 07°32.993’N, 008°24.753’W;

425 m asl). Four males measured 34.0—36.5 mm. In the

Yéalé village, a K. cochranae metamorph was found by

dip-netting in a deep pond in dense farmbush vegetation.

At night, adult males were heard calling at the same site

between inaccessible dense vegetation, edging a swamp

(07°31.928’N, 008°25.401’W; 425 m asl).

Amphib. Reptile Conserv.

Odontobatrachidae

Odontobatrachus arndti Barej, Schmitz, Penner,

Doumbia, Sandberger-Loua, Emmrich, Adeba, and

Rédel, 2015

Arndt’s Toothed Frog

Material: Three females, NGK-Nimba 0244, NGK-

Nimba 0245, NGK-Nimba 0246, and two males, NGK-

Nimba 0247 (Fig. 11A), NGK-Nimba 0248. Comments:

Odontobatrachus arndti is a torrent-frog living in

primary and slightly degraded forests, known from

Mount Sangbé and Mounts Nimba (Barej et al. 2015;

Channing and Rodel 2019). We found a few populations

of O. arndti along cascades of streams in forested ravines

edged by savannah (07°35.233’N, 008°25.190’W; 847 m

asl). These frogs were found to be very abundant along

a very torrent stream (07°34.652’N, 008°24.966’W; 716

m asl). In a lower part of the forest, an additional site

was along a wide torrent stream with a gravel bottom and

blocks of granite rock (07°33.121°N, 008°25.036’W; 422

m asl). The recorded males exhibited huge bright orange

femoral glands. They measured 45.1—52.5 mm (N = 8);

the female SUL ranged from 43.5—60.5 mm (N = 10),

thus the sizes of both sexes are within the known range

of the species (Bare] et al. 2015). At all sites, the majority

of frogs were close to the rocky streams, however, a few

females perched on trees close to the streams. These

records are the second for Ivory Coast.

Phrynobatrachidae

Phrynobatrachus alleni Parker, 1936

Allen’s Puddle Frog

Material: Male, NGK-Nimba 0010 (Fig. 11C).

Comments: During the rainy season, we found a yellow

Phrynobatrachus alleni male (18.5 mm) in breeding

condition, among leaf litter on the forest floor, in a patch

of dense forest, crossed by a small stream (07°32.993’N,

008°24.753’W; 425 m asl). Rodel (2003) reported from

nearby Mount Sangbé that breeding P. alleni became

completely yellow. The yellow color may disappear

within minutes when the frogs are disturbed. In Tai

National Park, breeding P. alleni males where never

observed to be completely yellow (M.-O. Rodel, unpub.

data).

Phrynobatrachus annulatus Perret, 1966

Ringed Puddle Frog

Material: Two females, NGK-Nimba 0091 (Fig. 11D),

NGK-Nimba 0094. Comments: Phrynobatrachus

annulatus 1s a forest-dwelling leaf litter frog, which has a

patchy distribution in forests from south-eastern Guinea,

eastern Liberia, western Ivory Coast, and western Ghana

(Ernst and Rédel 2006; Hillers and Rédel 2007; Rodel

et al. 2005; Rodel and Glos 2019). In Ivory Coast, the

species was reported from Tai National Park (e.g., Ernst

and Rodel 2006; Hillers et al. 2008c), and the Mabi- Yaya

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Kanga et al.

t jana

raeoe ag

Pid ‘iy

& ee,

Poo od

, gee,

it 5

aia

Fig. 11. Hyperoliid, odontobatrachid, and phrynobatrachid frogs from Mount Nimba Integrated Nature Reserve: Kassina cochranae

male (A); Odontobatrachus arndti male (B); Phrynobatrachus alleni male (C); P. annulatus male (D); P. francisci female (E); P.

fraterculus female (F); P. guineensis male (G); P. gutturosus female (H).

Amphib. Reptile Conserv. 89 June 2021 | Volume 15 | Number 1 | e275

Amphibians of the Nimba Mountains (Ivorian part)

Forest Reserve (Gongomin et al. 2019). We found two

young females of P annulatus (14.0 and 20.2 mm) in

a site with high canopy forest on the slopes of a large

granite inselberg. The dry forest ground was covered

with multiple layers of leaf litter, but the undergrowth

was sparse (07°34.364’N, 008°24.746’W; 643 m asl).

Drier parts of the forest along slopes of inselbergs

also comprise the usual habitat where this species was

recorded in Tai National Park (M.-O. Rodel, unpub.

data).

Phrynobatrachus francisci Boulenger, 1912

Francisc’s Puddle Frog

Material: Two females, NGK-Nimba 0092 (Fig. 11),

NGK-Nimba 0093. Comments: Phrynobatrachus

francisci occurs in moist Guinea savannah and drier

Sudanese savannah, from Senegal to Nigeria (Rodel

2000; Channing and Rédel 2019). We found the species

in the rainy season, among dense herbaceous vegetation

at the edge of a puddle (07°31.928’N, 008°25.401’W;

425 m asl). Two females (20.5—22.0 mm) were caught

in the Yéalé village during late afternoon (1618 h GMT).

Phrynobatrachus fraterculus (Chabanaud, 1921)

Brother’s Puddle Frog

Material: Female, NGK-Nimba 072 (Fig. I1I1F).

Comments: Phrynobatrachus fraterculus is known to

inhabit degraded forest and forest edges in the western

part of the Upper Guinea forest region (Guibé and

Lamotte 1963; Rodel and Bangoura 2004; Rédel and

Glos 2019). A female (24.0 mm) was found in leaf litter

near a shallow creek in a small clearing (07°33.121°N,

008°25.036’W; 422 m asl). In Ivory Coast, the species

had been recorded previously in the Tai National Park

(Ernst and Rodel 2006; Hillers et al. 2008c).

Phrynobatrachus guineensis Guibé and Lamotte, 1962

Guinea Puddle Frog

Material: Male, NGK-Nimba 0034 (Fig. 11G).

Comments: Phrynobatrachus guineensis is the only

known West African member of its genus that uses water-

filled tree holes as breeding sites (R6del 1998; Rodel et

al. 2004; Rudolf and Rodel 2007). A breeding male (15.5

mm) was found in a water-filled tree hole in a patch of

dense forest (07°34.696’N, 008°25.015’W; 717 m asl).

Phrynobatrachus gutturosus (Chabanaud, 1921)

Guttural Puddle Frog

Material: Two females, NGK-Nimba 0095 (Fig. 11H),

NGK-Nimba 0096, and two males, NGK-Nimba 0097,

NGK-Nimba 0098. Comments: A complex of cryptic

West African puddle frogs is currently known under the

name Phrynobatrachus gutturosus (Rodel 2000; Zimkus

et al. 2010). One species of that complex, P. afiabirago,

has been recently described from southern Ghana (Ofori-

Boateng et al. 2018). Frogs from this complex became

known from primary rainforest to dry savannah habitats

Amphib. Reptile Conserv.

(Rodel 2000; Rodel and Spieler 2000; Ernst and Rédel

2006; Nago et al. 2006; Hillers et al. 2008c), and their

taxonomic status requires further research. We found

P. gutturosus among leaf litter in a cocoa and coffee

plantation (07°31.928’N, 008°25.401’W; 425 m asl).

The plantation comprised a large pond and was used by

people from the Yéalé village to grow rice. Numerous

individuals of P. gutturosus were seen after sunset (1830

h GMT). Two females measured 18.5 and 20.0 mm.

Phrynobatrachus latifrons Ahl, 1924

Savannah Puddle Frog

Material: One female, NGK-Nimba 0064 (Fig. 12A),

and one male, NGK-Nimba 0065 (Fig. 12B). Comments:

Phrynobatrachus_ latifrons is a very common and

widespread, semi-aquatic West African puddle frog,

living in savannah and heavily degraded rainforest

habitats (R6del 1995 [there erroneously termed P

francisci], 2000; Kouamé et al. 2018). The species was

abundant in the Yéalé village. In particular, we heard

males calling between densely vegetated parts of swamps

and paddy fields (07°31.928’N, 008°25.401’W; 425 m

asl). An adult female with a broad green back and a light

vertebral line exhibited an unusual thin, light longitudinal

line on upper surface of the tibia (22.0 mm). A uniform

brown frog (18.1 mm) exhibited the bright yellow throat,

typical for adult males.

Phrynobatrachus liberiensis Barbour and Loveridge,

1927

Liberian Puddle Frog

Material: Male, NGK-Nimba 0071 (Fig. 12C).

Comments: Phrynobatrachus liberiensis is generally

found along swamps and shallow streams in primary

forests (R6del and Branch 2002; Kouamé et al. 2018).

Many males were heard calling, being well concealed

sitting on the banks of a shallow stream in a swampy

patch of dense forest (07°32.993’N, 008°24.753’W; 425

m asl). A male (27.0 mm) and a young female (26.0 mm)

were caught.

Phrynobatrachus natalensis (Smith, 1849)

Natal Puddle Frog

Material: Two males, NGK-Nimba 0032 (Fig. 12D),

NGK-Nimba 0033, and two females, NGK-Nimba

0035, NGK-Nimba 0036 (Fig. 12E). Comments:

Phrynobatrachus natalensis as_ currently defined

(Channing and Rodel 2019) comprise several cryptic

Species widespread throughout the savannah areas of

sub-Saharan Africa (Zimkus et al. 2010). We encountered

the species in the Yéalé village, near road puddles in

dense vegetation (07°31.928’N, 008°25.401’>W; 425 m

asl). Other active males were found at night between

tufts of ornamental plants around houses after heavy

rainfalls. Adult males were uniform brown (26.0—28.1

mm; N = 6) and had black throats with folds, while

adult females (27.8—33.5 mm; N = 8) had white, mottled

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Kanga et al.

ase yn —

ais eae a

pag eae

lel

tmor

eo ae ac me A ae Seis Fa f : .

Fig. 12. Phrynobatrachid frogs from Mount Nimba Integrated Nature Reserve: Phrynobatrachus latifrons female (A); P. latifrons

male (B); P. /iberiensis male (C); P. natalensis male (D); P. natalensis female (E); P. phyllophilus male (F); P. tokba female (G); P.

tokba male (H).

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Amphibians of the Nimba Mountains (Ivorian part)

brown or greyish-black throats. One female exhibited

an exceptionally conspicuous greyish-brown back with

green spots and a green interorbital line; other females

being uniform brown. Some frogs reproduced in a large

pond within dense vegetation. Four clutches comprising,

938, 1021, 1265, and 1501 small reddish-brown eggs,

were floating on the water surface. Mean egg diameter

was 0.9 mm (+ 0.1 mm; N = 20). Clutch sizes therefore

seem to be larger in West African, compared to southern

African, populations (compare values in Rédel 2000).

Phrynobatrachus phyllophilus Rédel and Ernst, 2002

Leaf-loving Puddle Frog

Material: Male, NGK-Nimba 0037 (Fig. 12PF).

Comments: Phrynobatrachus phyllophilus _ prefers

patches of swampy primary forest from eastern Ivory

Coast to Sierra Leone (R6del and Ernst 2002a; Ernst and

Rodel 2006; Kouameé et al. 2008, 2014, 2018; Channing

and Rodel 2019). We found only one male (15.0 mm)

during the rainy season. It was concealed among dense

leaf litter, in a patch of dense forest, intersected by a

small stream (07°32.993’N, 008°24.753’W; 425 m asl).

Phrynobatrachus tokba (Chabanaud, 1921)

Tokba Puddle Frog

Material: Female, NGK-Nimba 0038 (Fig. 12G).

Comments: Phrynobatrachus tokba occurs in primary to

degraded forests, and montane grassland, from western

Guinea to Ghana (Guibé and Lamotte 1963; Rodel

et al. 2004, 2005; Kouamé et al. 2018; Channing and

Rodel 2019). This species reproduces terrestrially, by

depositing clutches in moist leaves, and has non-feeding,

non-hatching tadpoles (R6del and Ernst 2002b). A large

number of P. tokba males called in the drier part of the

forest. A brown female measured 18.0 mm. Further

populations were detected in moist savannah adjacent

to forest, and even in sympatry with Nimbaphrynoides

occidentalis up to 1,235 m asl in montane grasslands

(07°35.555’N, 008°25.788’W; Fig. 12H).

Pipidae

Xenopus tropicalis (Gray, 1864)

Tropical Clawed Frog

Material: Two females, NGK-Nimba 0001 (Fig. 13A),

NGK-Nimba 0002, and two males, NGK-Nimba 0003,

NGK-Nimba 0006. Comments: This pipid lives in

forests, degraded forests, and gallery forests in humid

savannahs from Senegal to western Cameroon (Rodel

2000). The species was seen in flooded paddy fields in the

Yéalé village (07°31.928’N, 008°25.401’W; 425 m asl).

In the dry season, frogs were easily caught in patches of

the same shallow swamps. Some juvenile frogs measured

21.5-30.0 mm (N = 5), adult males reached 32.4-42.5

mm (N = 5), and adult females ranged from 48.3—53.3

mm (N = 4).

Amphib. Reptile Conserv.

Ptychadenidae

Ptychadena arnei Perret, 1997

Schiotz’s Grass Frog

Material: Male, NGK-Nimba 0053 (Fig. 13C).

Comments: Ptychadena arnei is a poorly studied frog

occurring in humid savannahs, secondary forests, and

gallery forests, from southern Senegal to central Ivory

Coast (Channing and Rodel 2019). Only two females

(43.5 and 45.5 mm) and two males (39.5 and 40.5

mm) were recorded at night in the Yéalé village, sitting

at the edge of a road puddle that intersected a heavily

degraded forest. The basic color of their back was brown

to grey with a light triangle on the snout. Ptychadena

oxyrhynchus, which often have a pale snout as well,

have much longer legs (see e.g., Fig. 13F). The P. arnei

specimens had short dorsolateral folds and possessed

distinct sacral folds. They had dark to pale crossbars

on the legs and lacked external metatarsal tubercles. A

female (Fig. 13B) exhibited a fine yellow vertebral line.

A reddish to yellow longitudinal line on the tibia was

broad towards the heel, but narrow towards the knee. The

upper part of flanks was reddish, while the lower part was

grey with black spots. The ventral surface ranged from

whitish through beige to yellow. The species is known

by its unique advertisement calls, which consist of a long

succession of short double calls (Channing and Rodel

2019). In paddy fields around Daloa, western-central

Ivory Coast, frogs that were morphologically identical

emitted such calls (Kouamé et al., unpub. data).

Ptychadena bibroni (Hallowell, 1845)

Bibron’s Grass Frog

Material: One female, NGK-Nimba 0104 (Fig. 13D),

and one male, NGK-Nimba 0105. Comments: We

found P. bibroni in the Yéalé village (07°31.928’N,

008°25.401’W; 425 m asl) along road puddles during

the rainy season, and in cocoa and coffee plantations

during the dry season. This frog is known from humid

savannahs, but was also reported from dry savannahs and

open degraded forests (R6del 2000; Channing and Rodel

2019). The body size of males ranged from 39.0-46.0

mm (N = 6), while females reached 46.5—57.0 mm (N

=7).

Ptychadena longirostris (Peters, 1870)

Snouted Grass Frog

Material: Female, NGK-Nimba 0039 (Fig. 136).

Comments: Pitychadena longirostris was found along a

forest trail with puddles of various sizes (07°32.993’N,

008°24.753’W; 425 m asl). They had a yellowish back

with an ill-defined darker lateral band, stretching from

the nares through eyes, tympanum, and flanks to the

groin. The size of males ranged from 43.0-46.0 mm (N

= 4); two females measured 50.0 and 52.5 mm (N = 2).

This West African species is known to breed in puddles

along forest roads (Guibé and Lamotte 1954; Rodel

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Kanga et al.

Fig. 13. Pipid and ptychadenid frogs from Mount Nimba Integrated Nature Reserve: Xenopus tropicalis female (A); Ptychadena

arnei female (B); P. arnei male (C); P. bibroni male (D); P. longirostris male (E); P. oxyrhynchus male (F); P. pujoli male (G); P.

pumilio female (H).

Amphib. Reptile Conserv. 93 June 2021 | Volume 15 | Number 1 | e275

Amphibians of the Nimba Mountains (Ivorian part)

2000). It preys on various insects and occasionally even

aquatic food items (Konan et al. 2016).

Ptychadena oxyrhynchus (Smith, 1849)

Sharp-nosed Grass Frog

Material: Two females, NGK-Nimba 0040, NGK-

Nimba 0041, and one male, NGK-Nimba 0042 (Fig.

13F). Comments: Pitychadena oxyrhynchus is a large

frog with extremely robust and long hind legs. It occurs

in savannahs and edges of the forest zone across sub-

Saharan Africa (Rodel 2000; Channing and Rodel 2019).

During the rainy season, the species was found in the

Yéalé village (07°31.928’N, 008°25.401’W; 425 m asl),

along dirt roads with temporary water bodies of different

sizes. The species survives the dry season in humid parts

of rice paddies. In this season, rice is harvested and we

saw the frogs in vast numbers. The sizes of two females

were 59.0 and 64.8 mm, while a male measured 52.0 mm.

Ptychadena pujoli (Lamotte and Ohler, 1997)

Pujol’s Grass Frog

Material: Male, NGK-Nimba 0045 (Fig. 13G).

Comments: The biology of Ptychadena pujoli is very

insufficiently known. It seems to occur in savannah

swamps and grassland habitats from eastern Sierra

Leone, through the Upper Guinea highlands, to western

Ivory Coast (Lamotte and Ohler 1997; Channing and

Rodel 2019). After a heavy rainfall, some migrating

individuals were found among short grasses near houses

in the Yéalé village (07°31.928’N, 008°25.401’W; 425

m asl). A male (48.5 mm SUL) with a brownish-grey

back had a beige vertebral band. Its back was smooth

to slightly granular. Flanks were light with some large

warts. The animal had continuous light-colored external

folds and distinguishable sacral folds. Its legs exhibited

greyish dark crossbars, and its feet lacked metatarsal

tubercles. The venter was yellowish. This species lived

in syntopy with P. arnei in rice paddies around villages in

the Daloa region (Kouamé et al., unpub. data).

Ptychadena pumilio (Boulenger, 1920)

Western Dwarf Grass Frog

Material: Female, NGK-Nimba 0069 (Fig. 13H).

Comments: A female P. pumilio was recorded in an open

area with a shallow but densely vegetated pond near

the Yéalé village (07°31.928’N, 008°25.401’W; 425 m

asl). The species 1s known from a wide range of habitats

from humid to dry savannahs, to open areas in degraded

forests, and ranges from southern Mauritania through the

savannah belt to eastern Africa (R6del 2000; Onadeko

and Rodel 2008; Padial et al. 2008; Adeba et al. 2010;

Channing and Rodel 2019).

Ptychadena retropunctata (Angel, 1949)

Nimba Grass Frog

Material: Two males, NGK-Nimba 0060, NGK-Nimba

0061, and two females, NGK-Nimba 0062, NGK-Nimba

Amphib. Reptile Conserv.

0063 (Fig. 14A). Comments: Five P. retropunctata

were recorded on a high plateau with predominantly

grassy mountain pastures and herbaceous vegetation

(07°35.453’N, 008°24.957°W; 843 m asl). The SULs

of males varied from 28.0—30.5 mm (N = 3), while two

females measured 29.0 and 37.5 mm. Whereas four frogs

were rust-colored, a female had a deep brown dorsum.

In contrast to Rédel (2000) and Channing and Rodel

(2019), who describe a white venter, our specimens had

a yellow venter. The species was described previously

from Mounts Nimba (Angel 1949). Further records

became known only from a few montane localities in

southeastern Guinea (summarized in Rédel et al. 2004),

the Loma Mountains in Sierra Leone (Lamotte 1971),

northern Guinea (Hillers et al. 2006), and southeastern

Senegal (Monasterio et al. 2016). This is the first country

record for P. retropunctata in Ivory Coast.

Ptychadena stenocephala (Boulenger, 1901)

Narrow-headed Grass Frog

Material: Two females, NGK-Nimba 0030, NGK-

Nimba 0100 (Fig. 14B). Comments: Two females

of the Ptychadena_ stenocephala-complex (Rodel

and Channing 2019) have been found at the edge

of a roadside puddle near a heavily degraded forest

in the Yéalé village (07°31.928’N, 008°25.401’W;

425 m asl). While they superficially resemble P.

mascareniensis, e.g., by the ridges and a similar

vertebral band (see e.g., R6del and Glos 2019), they

differ from the latter species by reduced webbing, and

much more slender body shape (Channing and Rédel

2019). The exact taxonomic status of these frogs

requires further research.

Ptychadena submascareniensis (Guibé and Lamotte,

1953)

Small Grass Frog

Material: Three females, NGK-Nimba 0044, NGK-

Nimba 0046, NGK-Nimba 0047 (Fig. 14C), and two

males, NGK-Nimba 0048, NGK-Nimba 0049 (Fig.

14D). Comments: We found a large population of P.

submascareniensis in predominantly grassy mountain

pastures with herbaceous vegetation (07°35.453’N,

008°24.957°W; 843 m asl). There, the species occurred

in vast numbers (> 1,500 individuals in an area of

approximately 50 ha) during the rainy season. Calling

males were observed during the day with peaks between

0830-1030 h GMT and 1830-2000 h GMT. The weather

conditions during these observations were characterized

by low visibilities due to mist and windy weather. We

recorded two migrating individuals in montane grassland

in sympatry with A. crusculum and N. occidentalis

(07°35.555’N, 008°25.788’ W; 1,235 m asl). These rarely

documented frogs (compare Guibé and Lamotte 1953,

1958a; Rodel and Bangoura 2004) have a compact body

with a moderately pointed snout. Adult females measured

29.5—32.0 mm (N = 8), while males ranged from 24.0-

June 2021 | Volume 15 | Number 1 | e275

Kanga et al.

= PR ce

oe ea

pn a

¥ Ps a

rf ee

i 7 — lee]

. ao

1 4 =,

; |

1 _ = t >

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= : ay 54 ~t Bs:<, 7 Tol ‘+

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Fig. 14. Ptychadenid, ranid, and rhacophorid frogs from Mount Nimba Integrated Nature Reserve: Ptychadena retropunctata (A),

P. stenocephala female (B); P. submascareniensis female (C); P. submascareniensis male (D); P. tournieri female (E); Amnirana sp.

‘albolabris west’ female (F); Amnirana sp. ‘albolabris west’ male (G); Chiromantis rufescens male (H).

Amphib. Reptile Conserv. 95 June 2021 | Volume 15 | Number 1 | e275

Amphibians of the Nimba Mountains (Ivorian part)

27.5 mm (N = 18). According to Channing and Rodel

(2019), Mount Nimba is the only known Ivorian site to

host P. submascareniensis.

Ptychadena tournieri (Guibé and Lamotte, 1955)

Tournier’s Grass Frog

Material: Two females, NGK-Nimba 0054, NGK-

Nimba 0055 (Fig. 14E), and one male, NGK-Nimba

0056. Comments: Piychadena tournieri 1s widespread

in patches of humid and dry savannahs from Senegal

into Benin (Rédel 2000; Nago et al. 2006; Adeba et al.

2010; Channing and Rédel 2019). We found the species

in open areas, most often among grasses at the edge of

plantations. One male measured 35.0 mm, while three

females ranged from 39.0—49.0 mm.

Ranidae

Amnirana sp. ‘albolabris-West’

West African White-lipped Frog

Material: Female, NGK-Nimba 0101 (Fig. 1I4F).

Comments: Amnirana sp. ‘albolabris-West’ is a forest-

dwelling frog occurring in the Upper Guinean zone,

formerly regarded as being conspecific with the real

A. albolabris from Central African forests (Jongsma et

al. 2018). While its description is being prepared, the

occurrence of this species has been even more recently

confirmed for Burkina Faso (Ayoro et al. 2020). This

taxon was recorded in the Yéalé village, where a chorus

was heard at night in a densely vegetated swampy area.

One to two males initiated calling and many other males

then joined the chorus. A similar behavior was also

observed in A. fonensis (Kouameé et al., unpub. data). In

the Yéalé village, frogs belonging to A. sp. ‘albolabris-

West’ were also found in large numbers (N > 50) in a

cocoa and coffee plantation near a large pond. Females

had a less spiny back than males (Fig. 14G).

Rhacophoridae

Chiromantis rufescens (Ginther, 1869)

Western Foam-nest Frog

Material: Male, NGK-Nimba 0102 (Fig. 14H).

Comments: Chiromantis rufescens is a complex

of treefrog species (Leaché et al. 2019), occurring

in rainforest and the transition zone between forest

and savannah, from West and Central Africa to the

southernmost record in Angola (Lamotte 1967; Schiotz

1967; Channing and Rodel 2019; Rédel and Glos 2019;

Ernst et al. 2020). They are the only West African species

depositing foam nests above stagnant waters (Coe 1967,

1974; Schigtz 1967; Monayong Ako’o 1978; Rodel et al.

2002). During the entire survey in Mounts Nimba, only

One specimen was found on a dark night in a plantation

from the Yéalé village dominated by cocoa and coffee

trees (Fig. 14H). This specimen measured 45.0 mm in

SUL.

Amphib. Reptile Conserv.

Discussion

The amphibian fauna of the Nimba Mountains ranks

among the best-known amphibian faunas in West Africa.

However, our knowledge is almost completely based

on records from the Guinean and Liberian parts of this

mountain range (e.g., Guibé and Lamotte 1958a,b, 1963;

Xavier 1978; Rédel et al. 2009, 2010; Sandberger et al.

2010; Sandberger-Loua et al. 2018a; Schafer et al. 2019).

Here, we present for the first time a comprehensive

summary of the amphibian fauna of the Ivorian part of

the mountains.

Such a summary Is particularly pressing as the Nimba

Mountains, with about 65 amphibian species recorded

so far, comprise the most species-rich amphibian site in

West Africa (summarized in Rédel et al. 2004; Barej et

al. 2015; Schafer et al. 2019; Channing and Rodel 2019;

and several unpublished records), but it is also highly

threatened by several factors. The Liberian part had been

mined already in the last century (Lamotte 1983), the

Guinean part has been prospected for mining and mining

will be implemented there soon (J. Doumbia, pers.

comm.). In the Ivorian part, natural forest and savannah

habitats in the lowlands are imperiled by agricultural

encroachment (Woods 2003), and mid-elevation and

mountain grasslands are prone to the uncontrolled

spread of bush-fires (Lamotte 1959; Hillers et al. 2008b;

Sandberger-Loua et al. 2016; this study). Thus, it is

important to understand the distributions of species, in

particular the endemic and threatened species, of the

amphibian fauna.

During our survey we detected 53 amphibian taxa.

The smooth skinned, mid-sized squeaker frogs of the

Arthroleptis poecilonotus-complex likely comprise

several species (R6del and Bangoura 2004), potentially

including A. nimbaensis. Species which are important

finds in this study are Nimbaphrynoides occidentalis,

Arthroleptis crusculum, Hyperolius nimbae, H. soror,

H. sp., and Kassina cochranae, as well as several

Ptychadena spp. (arnei, pujoli, retropunctata, and

submascareniensis). These species all have limited ranges

in the western part of the Upper Guinea biodiversity

hotspot, where at least two of them, Nimbaphrynoides

occidentalis and Hyperolius nimbae, are endemic to

the Nimba Mountains (Channing and Rodel 2019)—

the first to the montane grasslands and the second to

the Ivorian foothills of the mountain. The montane and

mid-elevation savannah grasslands of the MNINR are

the only known and remaining Ivorian habitats for N.

occidentalis, P. retropunctata, and P. submascareniensis.

Sandberger-Loua et al. (2016) have shown that N.

occidentalis comprises three isolated populations, two in

Guinea and one in Liberia, however, there 1s some gene

flow (Sanberger-Loua et al. 2018b). So far, it is unclear

if the Ivorian records comprise a fourth population

or (more likely) are sub-populations of the Guinean

populations. Their habitats, the montane grasslands, are

June 2021 | Volume 15 | Number 1 | e275

Kanga et al.

facing the uncontrolled spread of bush-fires. In lowland

habitats, human encroachments, forest fragmentation,

and steadily increasing agricultural activities, and thus

most likely a variety of agrochemicals, are crucial threats

for other amphibian species such as H. nimbae. However,

large areas of dense, broadleaf and evergreen forests,

stretching from the lower to mid-elevation slopes, still

prevail in MNINR (Fig. 4). Fourteen species, which

became known from the Guinean and Liberian parts

of the Nimba mountains, could not be detected in this

study: Arthroleptis krokosua, A. langeri, Sclerophrys

chevalieri, Afrixalus weidholzi, Hyperolius zonatus,

Phlyctimantis boulengeri, Phrynobatrachus calcaratus,

P._ hieroglyphicus, P. maculiventris, Ptychadena

superciliaris, P. aff. aequiplicata, Odontobatrachus

natator, Amnirana occidentalis, Geotrypetes

pseudoangeli, and G. seraphini. One further species,

Kassina lamottei, was reported previously from the

Ivorian Nimba area, but likewise could not be recorded

here. The latter species seems to require large pristine

lowland rainforests and reproduces in larger, stagnant

ponds (but not in rivers or puddles). In Tai National Park,

it was one of the most sensitive species concerning forest

alteration (Rédel and Ernst 2001a; Ernst and Rodel 2005;

Ernst et al. 2006).

For most of these species, our searching methods,

which included acoustic sampling, should be efficient,

as shown by our estimation curves indicating sufficient

sampling effort for the species richness that occurrs in

the Ivorian sector of Mounts Nimba. Additionally, many

species are more readily detected by their calls than

sight, and as frogs were not active at all times, most

of them are naturally rare and hide well, and thus are

difficult to detect. Some may have escaped our attention

in the field, e.g., tree-frogs hiding in the canopy. In order

to find caecilians, which mainly live underground, we

would have needed to dig in the most promising habitats.

However, it is absolutely possible that at least some of

these species are absent from the Ivorian slopes of the

mountain. For instance, Arthroleptis langeriis only known

from a few individuals of the Guinean part of Mount

Nimba and isolated populations in Liberia (Nopper et al.

2012), while Phrynobatrachus maculiventris has been

confirmed from a few sites in Guinea and Liberia (Rodel

et al. 2009), and P. hieroglyphicus 1s only known from

the type specimen and a locality in Liberia (Rodel et al.

2010). However, some other species, like Phlyctimantis

boulengeri, are known to occur even in disturbed habitats

(Rodel and Ernst 2001b, 2003), and thus were expected

to occur in MNINR as well. Likewise, our search for

Kassina lamottei in Zéalé, a formerly known Ivorian

lowland locality from the eastern flanks of Mounts

Nimba (Schietz 1967), remained unsuccessful (NG

Kouamé et al., unpub. data). There, primary forest has

been cleared completely and new asphalted concrete

roads facilitate commerce between Ivory Coast and

Guinea (NG Kouameé, pers. obs.).

Amphib. Reptile Conserv.

With 53 amphibian taxa, the MNINR is less diverse

compared to the known species records in the Guinean

part of the Nimba Mountains (Guibé and Lamotte

1958a,b, 1963; Xavier 1978; Rodel et al. 2009, 2010;

Sandberger et al. 2010; Sandberger-Loua et al. 2018a;

Schafer et al. 2019). However, the Guinean part has a

much larger area (12,540 ha) with a broader range of

forested habitats, thus representing the most significant

part of the Nimba mountain chain. As part of a set of

discontinuous montane reliefs, the Upper Guinea

mountain ranges extend along a northwest-southeast line

from the Fouta Djalon massif in western Guinea to the

regions of Danané and Man in Ivory Coast. The western

slopes of these mountains generally receive higher

precipitation than the eastern slopes (R6del and Bangoura

2004; Lauginie 2007; Sandberger-Loua et al. 2017). As a

consequence, the Ivorian slopes comprise more savannah

patches. This may also explain the restriction of H.

nimbae to the eastern slopes, as this reedfrog seems to

prefer the transition zone between forest and savannah

(Kouamé et al. 2016; this study). Nevertheless, when

comparing various amphibian assemblages, both Nimba

slopes share the highest faunal similarity in the region

(comparing Table 3 and Appendix). Compared with other

nearby sites, the amphibian species richness of MNINR

is also higher than those of Mount Sangbé National Park

(Rodel 2003), Diécké Forest Reserve (Rodel et al. 2004),

Ziama Forest (Chabanaud 1920, 1921; Bohme 1994a,

1994b), and the Pic de Fon-Simandou range (Parker

1936; Taylor 1968; Rodel and Bangoura 2004). Still,

MNINR shares a high number of amphibian species

(48-72%) with these sites. Interestingly two range

restricted species, e.g., Conraua cf. alleni (Fig. 8A—B)

and Odontobatrachus arndti (Fig. 11B), are known

from the Guinean part of Mounts Nimba (Barej et al.

2015; Schafer et al. 2019), the MNINR (this study),

and the adjacent Mount Sangbé National Park (Rodel

2003). This may indicate an area of unique amphibian

composition. For instance, the Conraua spp. from Mount

Sangbé, and potentially those from MNINR, differ in

call characteristics from those in more southern Ivorian

localities (R6del and Branch 2002; M.-O. Rodel, unpub.

data). The cryptic reedfrog, Hyperolius sp. (Fig. 10G-—

H), needs further investigation because it may represent

an undescribed species. Compared with other pristine

forest areas, the species richness of MNINR is in the

range of Tai National Park in southwestern Ivory Coast

(Roddel and Ernst 2004; Ernst et al. 2006; Table 3 and

Appendix). Although the Tai National Park is a lowland

habitat, it shares 62% of the amphibian species with our

study area, emphasising the high diversity of the Upper

Guinea rainforests. In addition to this richness of lowand

rainforest, the Nimba Mountains are thought to be a

refuge area where different faunas met and survived. For

instance, Arthroleptis krokosua is otherwise only known

from Ghana and Guinea, and in contrast Odontobatrachus

arnati and O. natator may be in contact with each other

June 2021 | Volume 15 | Number 1 | e275

Amphibians of the Nimba Mountains (Ivorian part)

on the southwestern slopes of the mountain. Some species

are endemic to the area (N. occidentalis, H. nimbae),

or seem to have their range center there: A. crusculum,

Kassina_ cochranae, Ptychadena_ retropunctata, P.

submascareniensis, P. arnei, and P. pujoli. The Nimba

Mountains are supposed to have represented a forest

refugium during dry Pleistocene times (Maley 1996),

thus forest species may have survived unfavorable

conditions for prolonged times in this area. Only with

strict protection of the Nimba Mountains and its habitats,

will such survival be possible in the future as well.

Conclusion

Our discoveries of Arthroleptis crusculum and

Ptychadena retropunctata added two species to the

list of the amphibian fauna of Ivory Coast. The unique

and diverse mosaic of habitats on Mounts Nimba

supports numerous species and their ecosystem services.

Our results should be taken as a baseline for future

management, monitoring, and conservation activities.

Most urgently, management efforts should try to mitigate

bush-fires in mountain grasslands, e.g., to protect the

viviparous Nimba toad, N. occidentalis. Furthermore, we

recommend the strict protection of the remaining forest

habitats. At lower elevations, it is important to encourage

local activities concerning reforestation of previously

forested areas and the conservation of (sacred) village

forests from which some rare or endemic (e.g., H. nimbae,

K. cochranae, P. arnei, P. pujoli), as well as threatened

species (e.g., Leptopelis macrotis) benefit.

Acknowledgements.—We are very indebted to the

“Ministere de |’Environnement et du Développement

Durable” of Cote dIvoire, and in particular the “Office

Ivoirien des Parcs et Réserves, Direction de Zone

Ouest” for permitting the research. We would like to

acknowledge the local populations of Danané villages

for their hospitality, and are especially grateful for

the support and collaboration from Paul Seu, elder of

Kouan-Houlé. We furthermore thank Nicolas Granier,

a conservationist of the Biotope Foundation France, for

facilitating relationships between the local people of

Yéalé and our team. We are also thankful to Droh David

Gueu, Berthé Zangbéli, and Zoda Alphonse Tokpa, our

local assistants and field guides, for their invaluable help

during the field work. We would like to acknowledge

Alan Channing and Raffael Ernst for their constructive

role as reviewers, improving the manuscript. We are

grateful to Werner Conradie for the valuable comments

he added regarding our manuscript.

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Kouassi Philippe Kanga is a Ph.D. candidate at the Universté Jean Lorougnon Guédé in Daloa,

Ivory Coast. His research topic focuses on amphibian diversity and its spatio-temporal dynamics in

the Ivorian part of Mount Nimba. Specifically, he aims to link amphibian assemblages with species

functions in this ecosystem, and to assess how species react in response to environmental changes.

N’Goran Germain Kouamé is an Ivorian herpetologist and biologist. He is senior lecturer and

head of the section biodiversity at the Université Jean Lorougnon Guédé, Daloa, Ivory Coast,

and the current Chair of the West African region of the IUCN SSC Amphibian Specialist Group

(ASG). He holds a Diploma and a Ph.D. in natural sciences from the Université d’ Abobo-Adjamé

(actually Université Nangui Abrogoua), Abidjan, Ivory Coast, where he used leaf-litter frogs

(Phrynobatrachus spp.) as models to determine the conservation status of the Banco National Park,

one of the rare remaining primary forests situated in the midst of a West African mega-city. His

current research interests mainly focus on the taxonomy, ecology, distribution, and conservation of

rare, threatened, and new amphibian species in Ivory Coast.

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Amphib. Reptile Conserv.

Amphibians of the Nimba Mountains (Ivorian part)

Parfait Zogbassé is a postgraduate of the Université Jean Lorougnon Guédé (Daloa, Ivory Coast)

with an M.Sc. in Herpetology. His current research focuses on the taxonomy, systematics, and

population ecology of grassfrogs (Ptychadena spp.) in western Ivory Coast. Parfait also teaches

natural sciences in a secondary school in Daloa as a public servant for the Ivorian government.

Basseu Aude-Inés Gongomin is a Ph.D. candidate at the Universté Jean Lorougnon Guédé (Daloa,

Ivory Coast). Her current research topic focuses on the taxonomy, DNA barcoding, and population

ecology of endemic amphibians of the Ivorian part of the Nimba Mountains.

Konan Laurent Agoh is a postgraduate student at the Université Jean Lorougnon Guédé (Daloa,

Ivory Coast) with an M.Sc. in Herpetology. His research interest focuses on the diversity and ecology

of amphibian assemblages in forest fragments of the valley of Bandama River, an area which lies

between Lamto Faunal Reserve (central Ivory Coast) and Banco rainforest (southern Ivory Coast).

Akoua Michéle Kouamé is a recent graduate of the Université Nangui Abrogoua (Abidjan, Ivory

Coast) with a Ph.D. in Herpetology. Her research focused on the life-history of reedfrogs in Azagny

National Park, a Ramsar site situated in southern-central Ivory Coast. Her research was supervised

by Professor Abouo Béatrice Adepo-Gourene and Dr. N’Goran Germain Kouamé.

Jean Christophe Béhibro YN Konan is a recent graduate of the Université Nangui Abrogoua

(Abidjan, Ivory Coast) with a Ph.D. in Herpetology. His research interests focus on the diversity,

taxonomy, systematics, ecology, and conservation of amphibians in Azagny National Park, a

Ramsar site situated in southern-central Ivory Coast. His research was supervised by Professor

Abouo Béatrice Adepo-Goureéne and Dr. N’Goran Germain Kouamé.

Abouo Béatrice Adepo-Gouréne is a professor at the Université Nangui Abrogoua (Abidjan,

Ivory Coast). Her research focuses on using animal genetics to better predict gene expression. She

is interested in the taxonomy and systematics of freshwater crustaceans, and the conservation of

Ivorian amphibians.

Germain Gouréne is the professor and founder of the “Laboratoire d’ Environnement et de Biologie

Aquatique” at the Université d’Abobo-Adjamé (actually Université Nangui Abrogoua, Abidjan,

Ivory Coast). His research focuses on the systematics and taxonomy of fishes, with an emphasis

on Africa; other areas covered include ecology and aquaculture. In addition to his research interest

on fishes, Germain is interested in the conservation of aquatic invertebrates and amphibians in the

Banco National Park. Germain has served as Vice-President and President of the University of

Abobo-Adjamé for 10 years, and was elected as deputy of the locality of Kounahiri (Central Ivory

Coast) from 2012-2016.

Mark-Oliver Rédel is the head of the department of “Diversity Dynamics” at the Museum fiir

Naturkunde, Berlin, Germany. He has studied the systematics, taxonomy, biogeography, and ecology

of African amphibians for almost 30 years. With his group, comprising students from around the

world, he currently runs projects in Germany, Guinea, Ivory Coast, Cameroon, Mozambique,

and Malaysian Borneo. His special interest is on how species and ecological communities react

to environmental changes. He has authored or co-authored more than 350 scientific publications,

including several books.

104 June 2021 | Volume 15 | Number 1 | e275

Kanga et al.

Appendix. Checklist of the amphibian species recorded in some mountain, highland, and lowland areas of the western part of the

Upper Guinean zones. Abbreviations: LF = Lowland Forest; LMEF = Lowland to Mid-Elevation Forest; DCF = Diéké Classified

Forest (R6édel et al. 2004); FD = Fouta Djalon Highlands (Hillers et al. 2008a; Barej et al. 2015); MB = Mount Béro (Rodel et al.

2004); MN (Guinea) = Mount Nimba in Guinea (Guibé and Lamotte 1958a, 1963; Laurent 1958; Schiotz 1967; Lamotte and Ohler

1997, 2000; Rodel et al. 2004; Barej et al. 2015; Sandberger-Loua et al. 2018; Schafer et al. 2019); MP = Mount Péko National

Park (Rédel and Ernst 2003); MS = Mount Sangbé National Park (Rodel 2003; Barej et al. 2015); SR = Simandou range (Parker

1936; Taylor 1968; Rédel and Bangoura 2004); TNP = Tai National Park (Rodel and Ernst 2004; Ernst et al. 2006); ZCF = Ziama

Classified Forest (B6hme 1994a,b; Chabanaud 1920, 1921). Literature records follow recent taxonomic changes.

Mountain and highland areas Mountain and highland areas LMEF

(this study)

Taxa

Dermophiidae

Geotrypetes pseudoangeli

Geotrypetes seraphini

Arthroleptidae

Arthroleptis crusculum

Arthroleptis krokosua

Arthroleptis poecilonotus-complex*

Astylosternus occidentalis

~*~

Cardioglossa occidentalis

Leptopelis bufonides

Leptopelis macrotis

Leptopelis occidentalis

Leptopelis spiritusnoctis

Leptopelis viridis

Bufonidae

Nimbaphrynoides occidentalis

Sclerophrys chevalieri

Sclerophrys maculata

Sclerophrys regularis

Sclerophrys taiensis

Sclerophrys togoensis x

Conrauidae

Conraua alleni x

Conraua sp. “fouta’

Dicroglossidae

Hoplobatrachus occipitalis x x

Hemisotidae

Hemisus guineensis x

Hemisus marmoratus xX

[ie a

om i el

a pes

Hyperoliidae

Acanthixalus sonjae D i ike %

gate: | ee Be) a

a a a |

[Atnanarigrenis «| «dE dT

ivcciawime Ed

| |

ae ———— ia ee

Amphib. Reptile Conserv. 105 June 2021 | Volume 15 | Number 1 | e275

~*~

Amphibians of the Nimba Mountains (Ivorian part)

Appendix (continued). Checklist of the amphibian species recorded in some mountain, highland, and lowland areas of the western

part of the Upper Guinean zones. Abbreviations: LF = Lowland Forest; LMEF = Lowland to Mid-Elevation Forest; DCF = Diéké

Classified Forest (Rédel et al. 2004); FD = Fouta Djalon Highlands (Hillers et al. 2008a; Bare] et al. 2015); MB = Mount Béro

(Rodel et al. 2004); MN (Guinea) = Mount Nimba in Guinea (Guibé and Lamotte 1958a, 1963; Laurent 1958; Schiotz 1967;

Lamotte and Ohler 1997, 2000; Rodel et al. 2004; Bare] et al. 2015; Sandberger-Loua et al. 2018; Schafer et al. 2019); MP = Mount

Péko National Park (R6del and Ernst 2003); MS = Mount Sangbé National Park (Rodel 2003; Bare] et al. 2015); SR = Simandou

range (Parker 1936; Taylor 1968; Rodel and Bangoura 2004); TNP = Tai National Park (Roédel and Ernst 2004; Ernst et al. 2006);

ZCF = Ziama Classified Forest (Bohme 1994a,b; Chabanaud 1920, 1921). Literature records follow recent taxonomic changes.

Mountain and highland areas rane || Mountain and highland areas LMEF

Taxa

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persis igtenenss =| «dT i

perciusiononsi + | x |_| | |

[ypersusninoae | x || +i

percucniaiee ————~|

persis aciaonte [i

[odontopatracitne iY

[odonatracinajoua ——————~

[Odonata aoe + id

[odonatacis mits | +t —*d

[odeneatacin tama | |_|

[Purwopatrachitne «YS S|

Panobanaencaten ____—+| | x — x [x]

Pinnobatachicamaons | x | |_| x | | _

Poach cotorans x

Pinpobatacmaprniucr |_| |

Pinnobatachupawraise | x |_|

Piobatachuguneenss | x ||

Pinnobatachu gute | x | x |x [x] |

Amphib. Reptile Conserv. 106 June 2021 | Volume 15 | Number 1 | e275

Kanga et al.

Appendix (continued). Checklist of the amphibian species recorded in some mountain, highland, and lowland areas of the western

part of the Upper Guinean zones. Abbreviations: LF = Lowland Forest; LMEF = Lowland to Mid-Elevation Forest; DCF = Diéké

Classified Forest (R6del et al. 2004); FD = Fouta Dyjalon Highlands (Hillers et al. 2008a; Barej et al. 2015); MB = Mount Béro

(Rodel et al. 2004); MN (Guinea) = Mount Nimba in Guinea (Guibé and Lamotte 1958a, 1963; Laurent 1958; Schiotz 1967;

Lamotte and Ohler 1997, 2000; Rodel et al. 2004; Bare] et al. 2015; Sandberger-Loua et al. 2018; Schafer et al. 2019); MP = Mount

Péko National Park (R6del and Ernst 2003); MS = Mount Sangbé National Park (Rodel 2003; Bare] et al. 2015); SR = Simandou

range (Parker 1936; Taylor 1968; Rodel and Bangoura 2004); TNP = Tai National Park (Rodel and Ernst 2004; Ernst et al. 2006);

ZCF = Ziama Classified Forest (Bohme 1994a,b; Chabanaud 1920, 1921). Literature records follow recent taxonomic changes.

(Mee

a an | 2 [om [oe [oe [os [ae

(this study)

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Ptychadena pujoli

Ptychadena retropunctata

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Anninanaspaboaivawet® | x | x |x [x | |x|x[x[x]x«_

[Riscophoriae

Amphib. Reptile Conserv. 107 June 2021 | Volume 15 | Number 1 | e275

Official journal website:

amphibian-reptile-conservation.org

Amphibian & Reptile Conservation

15(1) [Taxonomy Section]: 108-125 (e276).

urn:lsid:zoobank.org:pub:36C13E32-9F37-4D60-B309-0FC2357C50EB

A new species of the genus Gekko Laurenti, 1768 (Squamata:

Gekkonidae) from the Nicobar Archipelago, with an overview

of congeners from the Andaman and Nicobar Islands

1*§$.R. Chandramouli, 7G. Gokulakrishnan, 27C. Sivaperuman, and **L. Lee Grismer

'Department of Ecology and Environmental Sciences, School of Life Sciences, Pondicherry University, Puducherry 605014, INDIA *Zoological

Survey of India, Andaman and Nicobar Region Centre, Haddo, Port Blair 744102, INDIA *Herpetology Laboratory, Department of Biology, La

Sierra University, 4500 Riverwalk Parkway, Riverside, California 92515, USA

Abstract.—A comprehensive systematic review of the genus Gekko Laurenti, 1768 in the Andaman and Nicobar

archipelago was undertaken. The known members Gekko verreauxi Tytler, 1865 and Gekko (Ptychozoon)

nicobarensis (Das and Vijayakumar, 2009) are redescribed based on new material collected during this study.

The systematic status of the Gekko smithii (s. lat.) population from the southern Nicobar Islands was reassessed

and found to represent a distinct species. Based on morphological and morphometric distinction, this allopatric,

insular population is described as Gekko stoliczkai sp. nov. Notes on ecology, natural history, morphology, and

distribution are presented for all these species, with recommendations on their conservation status.

Keywords. Asia, cryptic species, endemic species, giant gecko, Reptilia, Sundaland

Citation: Chandramouli SR, Gokulakrishnan G, Sivaperuman C, Grismer LL. 2021. A new species of the genus Gekko Laurenti, 1768 (Squamata:

Gekkonidae) from the Nicobar Archipelago, with an overview of congeners from the Andaman and Nicobar Islands. Amphibian & Reptile Conservation

15(1) [Taxonomy Section]: 108-125 (e276).

[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: 1 April 2021; Published: 30 June 2021

Introduction

The nocturnal, arboreal members of the true gecko genus,

Gekko Laurenti, 1768, are currently represented by 77

species, which range from eastern India to Papua New

Guinea (Uetz et al. 2020). Wood et al. (2019) recently

partitioned Gekko into seven subgenera, including two

previously recognized genera, Ptychozoon Kuhl and

van Hasselt, 1822 and Luperosaurus Gray, 1845. The

Andaman and Nicobar Islands, situated to the east

of the Indian peninsula and south of the Ayeyarwady

Delta region of Myanmar, lie on the western periphery

of the geographic range of the genus Gekko. Currently,

this genus is represented by three species within the

Andaman and Nicobar archipelago, namely Gekko

verreauxi Tytler, 1865 from the Andaman Islands, Gekko

(Ptychozoon) nicobarensis (Das and Vijayakumar, 2009)

from the Nicobar Islands, and a species from the Nicobar

Islands currently identified as Gekko smithii Gray, 1842

(Uetz et al. 2020). Of these, Gekko verreauxi, the earliest

species known from the Andaman and Nicobar Islands,

was reported from the Andaman Islands as Gekko

gecko (Linnaues, 1758) as early as Blyth (1863). Gekko

verreauxi was once considered a synonym of G. stentor,

which again, was a junior synonym of G. smithii (fide

Boulenger, 1885) before being revived as a valid species

by Ota et al. (1991) based on an examination of type

material. This was followed by the report of Ptychozoon

homalocephalum|now Gekko (Ptychozoon) nicobarensis|

from the Nicobars by Steindachner (1867), which was

apparently overlooked by Das and Viyyakumar (2009).

The third species from this Archipelago, Gekko smithii,

was reported by Biswas and Sanyal (1977) from Great

Nicobar as Gekko gecko (in this study regarded as Gekko

cf. smithii), based on specimens at the Bombay Natural

History Society (BNHS) collected by Humayun A bdulali.

Subsequently, Biswas (1984) reported additional Gekko

smithi (sic) records from the South Andaman and Great

Nicobar Islands. Vesely (1999) provided additional notes

on the morphology and distribution of Gekko verreauxi.

Das (1999) speculated that the Nicobar population of G.

smithii was potentially allied to an undescribed member

of this group from Sumatra and specifically distinct from

the nominate species, Gekko smithii. Here, we examine

the three species of Gekko of the Andaman and Nicobar

Islands; provide additional data on the morphology,

*Correspondence. findthesnakeman@gmail.com, lgrismer@gmail.com

Amphib. Reptile Conserv.

June 2021 | Volume 15 | Number 1 | e276

Chandramouli et al.

natural history, and distribution of G. verreauxi and G.

(P). nicobarensis; and reassess the taxonomic status

of Gekko cf. smithii populations from the Nicobar

archipelago.

Materials and Methods

Specimens of Gekko encountered in the field were

gently restrained, measured, scored for morphological

characters (see below), photographed, and released at

the site of capture. Four specimens of G. verreauxi were

collected from the Andaman Islands, three specimens

of G. (Ptychozoon) nicobarensis were collected from

Car Nicobar, Camorta, and Teressa Islands, and five

specimens of the G. cf. smithii were collected from

Great and Little Nicobar Islands in and around human

habitations between 2014 and 2017. These specimens

were preserved in 70% ethanol and are deposited in

the two collections of the Zoological Survey of India,

Andaman and Nicobar Islands Regional Centre, Port

Blair, India (ZSI, ANRC) and the Department of Ocean

Studies and Marine Biology (DOSMB), Pondicherry

University, Brookshabad, Port Blair, India.

The following measurements (in mm) were recorded

with vernier calipers: Snout-vent length (SVL), measured

from the snout tip to the anterior edge of the cloaca;

tail length (TAL), measured from the posterior edge of

the cloaca to the tail tip; trunk length (AG), measured

between the axilla and groin; head length (HL), measured

from the snout tip to the angle of the jaw; head width

(HW), measured at the widest point on the head; head

depth (HD), measured from the top of the head to the

throat, at the level of the eyeball; horizontal eye diameter

(ED), eye-nostril distance (EN), measured from the

anterior margin of the eye to the posterior margin of

the external nares; snout length (ES), measured from

the anterior margin of the eye to the tip of the snout:

distance from eye to tympanum (ETY), measured from

posterior margin of the eye to the anterior edge of the

tympanum; tympanum diameter (TYD), measured across

the widest point of the tympanic opening; upper arm

length (UAL), measured from the axilla to the elbow;

lower arm length (LAL), measured from the elbow to

the wrist; palm length (PAL), measured from the wrist to

the tip of finger HI; thigh length (FEL), measured from

the point of insertion of the hind limb on the trunk to

the knee; tibia length (TBL), measured from the knee

to heel; foot length (FOL), measured from heel to the

tip of toe IV; and lengths of fingers (FI—F5) and toes

(T1-T5) measured from the fork of the digits to the

tip excluding the claws. Supralabials and infralabials

were counted along the upper and lower lips from

rostral and mental to the gape, respectively. Enlarged

scales bordering the mental posteriorly were counted as

postmentals, and expressed as inner pair + outer pairs.

Ventrals were counted along a transverse series across

the underside at mid-body between the ventrolateral

Amphib. Reptile Conserv.

folds; internasals were counted between the nasal scales;

subdigital lamellae were counted on the ventral surface

of digits of the fingers and toes, and dorsal tubercle rows

were counted in a transverse series at the mid-body. GPS

coordinates of the localities where the individuals were

encountered were recorded with a Garmin GPS MAP 78s

and mapped with ARC MAP v. 10. Comparative material

of the Sundaic G. smithii are provided in the Appendix.

The following analyses were carried out to test the

hypothesis that the Nicobar population of Gekko smithii

is morphologically distinct from those of the Sundaic

regions. A Student’s f-test was conducted on meristic

and mensural data (see below) with statistically similar

variances (1.e.,p values >0.05 inaLevene’stest)and normal

distributions (1.e., p values > 0.05 in a Shapiro-Wilk’s W

test) to search for the presence of statistically significant

mean differences (p < 0.05) among species across the

data set. Boxplots were generated for all characters in

order to visualize the range, mean, median, and degree

of differences between species pairs bearing statistically

different mean values. All statistical analyses were

performed in R (v3.4.3). The morphospatial clustering of

the sampled individuals was visualized using Principal

Component Analysis (PCA) from the ADEGENET

package in R (Jombart et al. 2010) implemented by

the prcomp() command. PCA is a dimension-reducing

algorithm that decreases the complexity of a data set

by finding a subset of input variables that contain the

most relevant information (i.e., the greatest variance in

the data) while de-emphasizing those characters that do

not, thus increasing the overall accuracy of the results

by eliminating noise and the potential for overfitting

(Agarwal et al. 2007). PCA is an unsupervised analysis

that recovers morphospatial relationships among the

sampled individuals (i.e., data points). It is important

to note that clusters of conspecific individuals were not

pre-defined in the analysis but simply color-coded in

the scatter plot in order to observe their positions and

morphospatial relationships. Meristic data were log-

transformed prior to analysis in order to normalize

their distributions, so as to ensure that characters with

very large and very low values could not over-leverage

the results owing to intervariable nonlinearity. To help

remove the potential effects of allometry in the mensural

characters, only adults were collected (as determined

by SVL). Additionally, variation in adult size was

normalized using the following equation: X,g~log(X)-

Bllog(SVL)-log(SVL,__..)], where X, = adjusted value;

X = measured value; B = unstandardized regression

coefficient for each population; and SVL... = overall

average SVL of all populations (Thorpe 1975, 1983;

Turan 1999; Lleonart et al. 2000; Tables 1 and 2). The

metrics of each species were normalized separately to

avoid conflating intra- with interspecific variation (Reists

1986). Meristic and mensural data were not concatenated

but analyzed separately owing to differences in sample

sizes. All data were scaled to their standard deviations

June 2021 | Volume 15 | Number 1 | e276

New Gekko species from Andaman and Nicobar Islands

Table 1. Summary statistics and Principal Component Analysis scores for the meristic characters.

PC1 PC2

Standard deviation 1.384 1.217

Proportion of variance 0.319 0.247

Cumulative proportion 0.319 0.566

Eigenvalue 1.916 1.480

Internasals -0.223 0.524

Infralabials -0.578 -0.388

Supralabials -0.633 -0.265

Postmentals -0.399 0.417

4" toe lamellae -0.153 0.026

Ventrals -0.182 Oo7e5

to insure they were analyzed on the basis of correlation

and not covariance. A Discriminant Analysis of Principal

Components (DAPC) was performed on both data sets.

The DAPC places individuals from each predefined

population into separate clusters (1.e., plots of points)

bearing the smallest within-group variance that produce

linear combinations of centroids having the greatest

between-group variance (1.e., linear distance; Jombart et

al. 2010). DAPC relies on scaled data calculated from

its own PCA as a prior step to ensure that the variables

analyzed are not correlated and number fewer than the

sample size. Dimension reduction of the DAPC prior

to plotting was accomplished by retaining the first set

of PCs that accounted for approximately 90% of the

variation in the data set (Jombart and Collins 2015) as

determined from a scree plot generated as part of the

analysis. Retaining too many PCs forces false structure

to appear in the data, while retaining too few runs the risk

of missing true structure (Cangelosi and Goriely 2007).

Additional museum abbreviations are: NHMUK

— The Natural History Museum, London (formerly,

BMNH - British Museum of Natural History), FMNH

— Field Museum of Natural History, Chicago, Illinois,

PC3 PC4 PCS PC6

0.994 0.860 0.813 0.463

0.165 0.123 0.110 0.036

0.731 0.854 0.964 1.000

0.988 0.740 0.661 0.214

-0.070 -0.810 0.111 -0.050

-0.077 -0.013 0.225 -0.678

-0.138 -0.026 0.019 0.714

0.056 0.281 -0.748 -0.158

0.974 -0.008 0.155 0.057

-0.136 0.513 0.594 0.029

USA; USNM -— National Museum of Natural History,

Washington, DC, USA; THMNH -— Thailand Natural

History Museum; LSUHC — La Sierra University,

Herpetology Collection; ZMA -— Universiteit van

Amsterdam, Zoologisch Museum, Amsterdam,

Netherlands; ZRC: Zoological Reference Collection,

Department of Zoology, University of Singapore.

Results

The results of the analyses support our initial hypothesis

that the Nicobar and Sundaic populations of Gekko

smithii are morphologically distinct. The PCA of

the meristic data show that the Nicobar population

overlaps with the Sundaic Gekko smithii along principal

component (PC) | but is generally well-separated from

Sundaic G. smithii along PC 2 (Fig. 1). PC1 accounts

for 31.9% of the variation in the data set and loads

most heavily for infralabials and supralabials (Table

1), while PC2 accounts for 24.7% of the variation and

loads most heavily for postmentals and internasals. Four

PC eigenvalues and the first discriminate function were

retained for the DAPC, which accounted for 92.9% of the

Table 2. Summary statistics and Principal Component Analysis scores for the mensural characters. Abbreviations are defined in the text.

PCI PC2 PC3 PC4 PCS PC6 PC7 PC8 PC9 PC10

Standard deviation 2.713. 0.930 0.745 0589 0526 0442 0364 0348 0318 0.221

Proportion of variance 0.736 0.087 0.056 0.035 0.028 0.020 0.013 0012 O.0O10 0.005

Cumulative proportion 0.736 0.822 0.878 0912 0.940 0.960 0.973 0.985 0.995 1.000

Eigenvalue 7.358 ‘0;865 0555 0347 0276 O0195> 0.132. (07121 0.101 0.049

HD 0.322 -0.250 -0.109 0.009 0.738 -0.011 -0.071 0.165 -0.426 = -0.251

HL 0.340 0.157 -0.035 -0.162 -0.329 0.034 -0.776 -0.117 -0.144 -0.292

HW 0.324 -0.220 -0385 -0.359 0.092 0.136 0.107 -0.046 0.701 ~~ -0.195

TYD 0.316 -0.297 -0.033 0459 -0.173 -0.741 -0.002 -0.010 0.144 0.002

EN 0.320 -0270 -0.117 0381 -0404 0529 0.336 -0.091 -0.260 -0.182

ES 0.356 -0.103 -0.055 0.023 0.023 0.208 -0.226 0.259 0.025 0.835

ED 0.208 0.777 -0447 0.336 0.146 -0.021 0.127 -0.022 0.041 0.010

UAL 0.285 0.179 0.743 0.281 0.229 0.210 -0.034 -0.052 0.374 = -0.122

LAL 0.340 0.088 0.147 -0.372 0.033 -0.171 0.280 -0.707 -0.236 0.230

AG 0.327 0.222 0.215 -0.398 -0.265 -0.181 0.348 0615 -0.144 -0.127

Amphib. Reptile Conserv.

June 2021 | Volume 15 | Number 1 | e276

Chandramouli et al.

PCA eigenvalues

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smithii -

Fig. 1. Analyses of meristic characters. (A) First two principal components of the PCA. (B) Density curves of the first discriminant

function of the DAPC. (C) Boxplots in which light-blue circles are the means and the black horizontal bars are the medians.

variation and demonstrated very little overlap between

the Nicobar population and G. smithii. These results are

mirrored by the mensural data, where the PCA of the

mensural data show that the Nicobar population and

Gekko smithii plot completely separately from each other

(Fig. 2). PC1 accounts for 73.6% of the variation in the

data set and loads fairly evenly for all characters, except

for low loadings for the eye diameter and forelimb length

(Table 2). PC2 accounts for 8.8% of the variation and

loads most heavily for eye diameter. Six PC eigenvalues

and the first discriminate function were retained for the

DAPC, which accounted for 97.1% of the variation and

demonstrated no overlap between the Nicobar population

and G. smithii. Student f¢-tests also showed several

statistically significant mean differences between the

Nicobar population and Gekko smithii in both meristic

and mensural characters (Table 3; Figs. 1—2).

Based on the data presented above, we believe there

is sufficient evidence to formulate a robust, testable

hypothesis indicating the Nicobar population is a distinct

species, and as such, 1s described below. In addition, we

redescribe the species G. verreauxi and G. nicobarensis

based on our new data.

Amphib. Reptile Conserv.

111

Taxonomy

Gekko (Gekko) verreauxi Tytler, 1865 (Figs. 3—4)

Gekko verreauxi Tytler, 1865

Gekko stentor — Boulenger (1885) part

Gekko smithii — Smith (1935) part

Table 3. Results of Student f-tests.

p-value t-value

Mensural characters

Head length 1.71E-06 -5.1

Head width 0.02105 -2.3457

Snout length 0.02042 -2.3577

Eye diameter 1.65E-10 -7.1575

Forelimb length 0.01159 -2.5737

Tibia length 2.13E-05 -4.4714

Axilla-groin length 8.11E-07 -5.2777

Meristic characters

Internasals 8.40E-07 -5.2692

Infralabials 2.23E-02 2.3234

Supralabials 0.028 2251 |

Ventrals 0.003089 -3.0357

June 2021 | Volume 15 | Number 1 | e276

New Gekko species from Andaman and Nicobar Islands

PCA eigenvalues

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function of the DAPC. (C) Boxplots in which light-blue circles are the means and the black horizontal bars are the medians.

Lectotype. NHMUK 68.4.3.64, an adult female collected

from the ‘Andaman Islands’ by W. Theobald fide Ota et

al. (1991) (photographs studied).

Paralectotype. NHMUK 1868.4.3.65, an adult female

from the same location (photographs studied).

Material studied. ZSI/ANRC/T/4324 from Mayjarpahar

(11.7029°N, 92.6433°E, 16 masl), South Andaman; ZSI/

ANRC/T/3726 from Durgapur (13.2761°N, 93.0311°E,

5 m asl), North Andaman; ZSI/ANRC/T/4566 from

Rabindranagar (10.7080°N, 92.5334°E, 57 m_ asl),

Little Andaman; ZSI/ANRC/T/5779 from Krishna

Nagar (12.0086°N, 92.9635°E, 58 m asl), Swarajdweep

(formerly Havelock Island).

Diagnosis. A large-bodied species of Gekko (SVL 140.06—

146.48 mm) characterized by: 12—14 supralabials; 9-13

infralabials; two elongate inner pairs of postmentals in

Amphib. Reptile Conserv.

112

broad contact with each other; two smaller, separated

outer pairs of postmentals; 11-13 precloacal pores in

males; absence of femoral pores; two internasals in

contact with each other; distinct ventrolateral dermal

folds; 26—29 transverse rows of imbricate ventrals; 12 or

13 transverse rows of enlarged, rounded, dorsal tubercles;

two to three enlarged post-cloacal spurs on each side

of the vent; 18-20 undivided subdigital lamellae on

toe IV; presence of five legible, dark, transverse bands

in juveniles and subadults transforming into a nearly

uniform dark-brown dorsal coloration in adults with a

pale-white to light-brown venter.

Description and variation. A large species of Gekko,

measuring 109.55-146.49 mm SVL. Head large (mean

HL:SVL 0.26), longer than broad (mean HL:HW 1.19)

with a blunt, rounded snout tip. Eyes moderately large

(mean ED:HL 0.25) with a vertically elliptical pupil

approximately one-half the length of the snout (mean

ED:ES 0.53); nostrils situated closer to the snout tip

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Chandramouli et al.

than to the eyes (mean EN:ES 0.79). Trunk one-half as

long as the body (mean AG:SVL 0.46). Overall habitus

depressed. Supralabials 12—14 on each side; infralabials

9-13 on each side; two moderately enlarged postmentals

in broad medial contact, followed by three smaller

pairs of scales. Dorsum bearing 11-13 transverse rows

of enlarged tubercles. Ventrals imbricate, in 27—29

transverse rows at midbody. A pair of enlarged, rounded

cloacal spurs at the base of the vent. Anterior subcaudals

not enlarged, posterior ones slightly enlarged. Upper arm

shorter than lower arm (mean UAL:LAL 0.79); palm

enlarged with five fingers; the first one with an indistinct

Amphib. Reptile Conserv.

Fig. 3. Gekko verreauxi in life from South Andaman (top, middle, and bottom right: adult; bottom left: juvenile).

claw; relative lengths of fingers IV>IN>V>II>I. Thighs

short (mean FEL:SVL 0.16), robust bearing a few

tuberculated scales on the dorsal surface. Tibia as long

as the thighs (mean TBL:SVL 0.16); toes with entire,

undivided subdigital lamellae, 18—20 on toe IV; relative

lengths of toes IV>III>V>H>I. Fingers and toes free,

lacking membranous skin flaps. Males have a series of

11-13 precloacal pores and lack femoral pores.

Coloration in life. Overall dorsal coloration uniform

dark- to light-brown with five to six darker transverse

bands on the body and five or six on the tail. Venter

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New Gekko species from Andaman and Nicobar Islands

Ned N

_ ‘

uniform brown and lacking any specific pattern. Eyes

with vertically elliptical pupils and a greenish yellow iris.

Distribution. Occurs throughout the Andaman Islands,

and was recorded from the following islands during the

present study: South, Middle, North and Little Andaman,

Rutland Island, Ritchie’s archipelago (Havelock, Neil,

John Lawrance, and Henry Lawrance), Kyd, Labyrynth

archipelago (Tarmugli, Alexandra, Hobday, and

Redskin), Interview, North Reef, Paget, Long, Guitar,

and North Passage Islands (Fig. 10).

Natural history. Nocturnal in habit. Found on tall trees

at a height of approximately 5 m and above the ground

in evergreen forests. It has also been recorded from

secondary forests, human habitations, and littoral forests.

Calls comprise of a series of repeatedly uttered, rattling

syllables of tuk-tuk-tuk-tuk.

Remarks. Boulenger (1885) synonymized Gekko

verreauxi under Gekko stentor (Cantor, 1847), a junior

synonym of Gekko smithii Gray, 1842. Ota et al. (1991),

while revalidating G. verreauxi from the synonymy

of G. smithii, provided a detailed redescription of the

species with measurements and scale-counts from

both the lectotype and the paralectotype. Both of these

specimens are females and hence, we have supplemented

Amphib. Reptile Conserv.

Fig. 4. Dorsal, ventral, and lateral views of the head and ventral view of the foot of Gekko verreauxi (ZSI ANRC T 4566).

the available information with new specimens, including

males, in order to improve our understanding of the

species. The lectotype has 14 supralabials (12-14 in

our material), 10 infralabials (9-13), 11 rows of dorsal

tubercles at midbody (11-13), and 21 subdigital lamellae

(1 8—20) under toe IV.

Gekko (Ptychozoon) _ nicobarensis

Vijayakumar, 2009) (Figs. 5—6)

Ptychozoon homalocephalum — Steindachner (1867)

Ptychozoon kuhli — Stoliczka (1870); Smith (1935);

Biswas and Sanyal (1977); Das (1999) part

(Das and

Holotype. ZSI 2603, an adult male collected by A.R.

Anderson from ‘Nicobars’ in the late 1800s.

Material studied. DOSMBO05102 from Chuckchuka

(9.2161°N, 92.8109°E, 14 m asl), Car Nicobar, ZSI/

ANRC/T/5234 from Kalasi (8.2803°N, 93.1173°E),

Teressa, and ZSI/ANRC/T/4235 from Chota Enaka

(8.0657°N, 93.5428°E, 39 m asl), Camorta, Nicobar

Islands.

Diagnosis. A medium to large-bodied (SVL 78.2—95.8

mm) species of Gekko, characterized by: presence of

extensive, membranous skin flaps along the sides of

the head, trunk, and tail; tail tip with an oval shaped

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Chandramouli et al.

Fig. 5. Gekko (Ptychozoon) nicobarensis in life from Car Nicobar (above: banded; middle: striped morphs).

skin flap; presence of 10-13 supralabials; 10 or 11

infralabials; two relatively small, elongate inner pairs

of postmentals in broad medial contact with each other;

three smaller, separated outer pairs of postmentals:

20-21 precloacal pores in males, no femoral pores; two

internasals in contact with or separated by one or two

small azygous scales; no ventrolateral body folds; 29-32

transverse, juxtaposed rows of ventrals; four well-spaced,

transverse rows of enlarged, rounded dorsal tubercles;

one enlarged post-cloacal spur on each side of the vent;

13-21 undivided subdigital lamellae on toe IV; dorsal

color pattern consisting of four or five legible, dark,

W-shaped, transverse bands or a light-colored vertebral

Amphib. Reptile Conserv.

stripe bounded on either side by two darker stripes, and a

creamy white to light-brown venter.

Description and variation. A medium to large-bodied

species of Gekko belonging to the subgenus Ptychozoon

measuring 78.42—95.86 mm SVL, head large (mean

HL:SVL 0.28), much longer than broad (mean HL: HW

1.34) with a blunt, rounded snout tip. Eyes protruding,

fairly large (mean ED:HL 0.25) with a vertically elliptical

pupil; eye nearly half as long as the snout (mean ED:ES

0.53); nostrils situated closer to the snout tip than to

the eyes (mean EN:ES 0.77). Trunk much shorter than

one-half the length of the body (mean AG:SVL 0.47).

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New Gekko species from Andaman and Nicobar Islands

Overall habitus depressed. Supralabials 10-13 and

infralabials 10—11 on each side; two moderately enlarged

postmentals in broad medial contact, followed by three

pairs of scales that are nearly as large as the postmentals.

Dorsum bearing four transverse rows of weakly

enlarged, rather indistinct tubercles. Ventrals imbricate,

in 29-32 transverse rows. Sides of the body bearing a

membranous flap of skin, with enlarged, rhomboidal,

imbricate scales. A pair of enlarged, rounded cloacal

spurs present at the base of the vent. Subcaudals not

elongated horizontally, the middle row slightly enlarged.

Tail with serrated dermal membranes extending along

the sides, forming an oval-shaped disc at the terminus.

Tail tip regenerated. Upper arm shorter than lower arm

(mean UAL:LAL 0.91); palm enlarged with five fingers;

the first one with an indistinct claw; relative lengths of

fingers IV>IN>V>I>I. Sides of upper and lower arms

with extensive skin flaps. Thighs short (mean FEL:SVL

0.18) robust; with a few tuberculated scales on the dorsal

surface. Tibia shorter than thighs (mean TBL:SVL 0.16);

toes with entire, undivided subdigital lamellae, 14-20

under toe IV; relative lengths of toes [V>IN>V>II>I. Tibia

bearing lateral flaps of skin. Fingers and toes extensively

webbed with membranous skin flaps extending from the

base to the tip. Males have a series of 20—21 precloacal

pores and lack femoral pores.

Coloration in life. Dorsal coloration variable. Two

distinct morphs recorded. One morph with a series of

Amphib. Reptile Conserv.

Pca

Fig. 6. Dorsal, lateral, and ventral views of the head and ventral view of the feet of Gekko nicobarensis (DOSMB05102).

five or six darker brown *‘W’ shaped transverse bands on

the body and five or six similar transverse bands on the

tail. The other morph with a bright tan vertebral stripe

bordered by darker flanks (Fig. 5). Venter uniform brown

and lacking any specific pattern. Eyes with vertically

elliptical pupils, a greenish yellow iris and brown

reticulations.

Distribution. Occurs on islands of the Northern and

central group of the Nicobar Archipelago. Recorded

from Car Nicobar, Camorta, Katchall, and Teressa

Islands during this study. It has been reported from

Chowra, Bompoka, Trinkat, and Nancowry Islands (Das

and Vijayakumar 2009) (Fig. 10).

Natural history. Nocturnal in habit. The individuals

recorded during this study were observed mostly at a

height of approximately 2—2.5 m above the ground on the

trunks of relatively short shrubs that were nearly 3—3.5 m

tall, with a girth not exceeding 25 cm. Found in primary

evergreen forests, also recorded from secondary forests,

human habitations, and plantations. Calls of this species

could not be heard during the sampling sessions in the

field. Pairs of eggs stuck to each other and the tree bark

substrate were seen in July.

Remarks. The holotype has 11 supralabials (10-13 in

our material), 9 infralabials (10-11), four rows of dorsal

tubercles at midbody (4), and 21 subdigital lamellae (14—

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Chandramouli et al.

Fig. 7. Gekko stoliczkai sp. nov. in life from Great Nicobar.

19) under toe IV.

Gekko stoliczkai sp. nov. (Figs. 7-9)

Gekko gecko (non Linnaeus, 1758) — Biswas and Sanyal

(1977) part

Gekko smithii (non Gray, 1841) — Biswas (1984); Das

Amphib. Reptile Conserv.

(1999); Vijayakumar (2005) part

urn:|sid:zoobank.org:act: 1795181 F-C7A D-4195-A 233-2B2F1B13826B

Holotype. ZSI/ANRC/T/6092, an adult male collected

from Makachua (7.4035°N, 93.7134°E, 37 m asl), Little

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New Gekko species from Andaman and Nicobar Islands

Fig. 8. Holotype (ZSI/ANRC/T/6092) of Gekko stoliczkai sp. nov. in dorsal (above) and ventral (below) views.

Nicobar Island on 14 August 2018 by G. Gokulakrishnan.

Paratypes. DOSMB05020, an adult female from

Shastri Nagar (6.8065°N, 93.8882°E, 41 m asl), ZSI/

ANRC/T/6093 an adult female, ZSI/ANRC/T/4796 and

ZSI/ANRC/T/7221, two adult males from East-West

road (7.0022°N, 93.8811°E, 82 m asl), Great Nicobar.

Etymology. The specific epithet is a patronym honoring

Dr. Ferdinand Stoliczka (1838-1874) for his early

contributions to the herpetology of Andaman and

Amphib. Reptile Conserv.

Nicobar Islands. Some of his works, such as Stoliczka

(1870; 1873), provided significant information on the

herpetofauna of the Andaman and Nicobar Islands,

which included the description of several new taxa such

as Rana gracilis var. andamanensis (now Minervarya

andamanensis), Rana gracilis var. nicobariensis (now

Minervarya nicobariensis), Hylorana nicobariensis

(now Bijurana nicobariensis), Typhlops andamanensis

(now Gerrhopilus andamanensis), Ablabes nicobariensis

(now Gongylosoma _nicobariensis), and Mocoa

macrotymapnum (now Lipinia macrotympanum).

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Chandramouli et al.

Table 4. Meristic and morphometric characters of specimens examined for (A) Gekko verreauxi, (B) Gekko nicobarensis, and (C)

Gekko stoliczkai sp. nov. The specimen number of the new species holotype is in bold.

Table 4A. Data for Gekko verreauxi.

Species Gekko verreauxi Gekko verreauxi Gekko verreauxi Gekko verreauxi Mean

Catalogue Number: ZSI/ ANRC/T/3726 = ZSI/ ANRC/T/5779_ ~— ZSI/ ANRC/T/4324 = ZSI/ ANRC/T/4566

Island North Andaman Havelock South Andaman Little Andaman

SVL (mm) 140.06 109.55 140.62 146.49 134.18

Trunk length 67.24 41.99 69.84 66.58 61.41

Tail 93.19 105.89 142.96 127.96 117.50

Head length 33.41 31.83 36.04 38.52 34.95

Head width 25.96 27.82 32.16 31.79 29.43

Head depth 17.14 L381 15.89 16.96 15.33

Eye diameter 8.84 9.11 8.02 8.29 8.57

Tympanum diameter 4.13 4.06 a 4.52 4.12

Eye-nostril 13.69 9.87 13.04 14.17 12.69

Eye-snout 16.18 13.86 15.88 18.66 16.15

Eye-tympanum 11.12 10.11 12.25 13.2 11.67

Supalabials 12 12 14 14 —

Infralabials 9 10 1] 13 —

Post-mentals 2+4 2+4 2+4 2+4 —

Ventrals 28 ap 28 29 —

Internarial distance | 5.09 5.26 Sale —

Upper arm length 14.35 12.31 15.53 16.23 14.61

Lower arm length 1562 16.03 20.76 21.56 18.49

Palm length 17.93 14.79 17.44 18.98 17.29

Femur length 18.52 18.71 25.24 24.16 21.66

Tibia length 20.75 20:72 21.08 25,53 22.03

Foot length 16.82 17.41 21.38 21.1 19.18

T4 lamellae 20 18 20 20 —

Sex M M F F —

Table 4B. Data for Gekko nicobarensis.

Species Gekko nicobarensis Gekko nicobarensis Gekko nicobarensis Mean

Catalogue Number: DOSMB05102 ZSI/ANRC/T/5234 ZSI/ANRC/T/4235

Island Car Nicobar Teressa Camorta

SVL (mm) 91.21 95.86 78.42 88.50

Trunk length 33.74 45.34 44.65 41.24

Tail 61 63.18 61.57 61.92

Head length 25.14 27.26 DE OS 25.02

Head width 17.45 20.38 18.33 18.72

Head depth 10.07 1227 11.02 11.26

Eye diameter 555 7.13 5.76 6.15

Tympanum diameter ZS 3.14 2.4 2.43

Eye-nostril 8.21 943 8.91 8.75

Eye-snout 10.54 11.94 11.54 11.34

Eye-tympanum 7.74 8.87 6.71 LT

Supalabials 10 13 12 —

Infralabials 10 1] 10 —

Post-mentals 2+6 2+6 2+6 —

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New Gekko species from Andaman and Nicobar Islands

Table 4B (continued). Data for Gekko nicobarensis.

Mean

Species Gekko nicobarensis Gekko nicobarensis Gekko nicobarensis Mean

Ventrals 29 32 A =

Internarial distance 4 4.46 3.45 =

Upper arm length 1S a 8.15 9:9') 9.74

Lower arm length 10.54 10.36 11.16 10.69

Palm length 10.11 O77 $27 9.38

Femur length 15.74 15.8 15.1 Je yni

Tibia length 14.58 15.55 13.14 14.42

Foot length 12.97 12.92 11.88 12.59

T4 lamellae 17 14 19 =

Sex F F F —

Table 4C. Data for Gekko stoliczkai sp. nov.

Species Gekko stoliczkai sp. Gekko stoliczkai Gekko stoliczkai Gekko stoliczkai Gekko stoliczkai

nov. Sp. nov. Sp. nov. sp. nov. Sp. nov.

Catalogue Number: ZSI/ANRC/T/6092. =DOSMB05020 ZSI/ANRC/T/6093 ZSI/ANRC/T/4796 ZSI/ANRC/T/7221

Island Little Nicobar Great Nicobar Great Nicobar Great Nicobar Great Nicobar

SVL (mm) 118.37 122 123.83 128.4 116.29

Trunk length 55.33 51.85 58.2: 60.64 49.08

Tail 105.9 112 130.49 81.31 103.81

Head length 32.1 31.6 30.22 36.93 30.43

Head width 24.04 23.43 21.46 25.07 23.19

Head depth 14.17 14.28 12.93 14.54 13.69

Eye diameter 7.84 6.76 8.05 8.66 7.68

Tympanum diameter 43 3,12 3.29 a ats) 2.63

Eye-nostril 11.77 11.16 1215 13.94 11.54

Eye-snout 15.04 14.05 14.95 FANS 14.51

Eye-tympanum 10.76 9.76 10.22 11.16 9.96

Supalabials 14 14 14 17 15

Infralabials 13 12 12 13 12

Post-mentals 2+4 2+4 2+4 2+4 2+4

Ventrals 22 25 23 21 22

Internarial distance 3.61 3 4.47 4.75 4.39

Upper arm length 11.28 16.99 12.46 13.12 12.28

Lower arm length 13.84 15.9 17.68 16.02 15.71

Palm length 15.54 16.83 12.84 16.15 11.29

Femur length 19.9] 20.96 18.88 20.96 21.15

Tibia length 18.19 20.06 17.78 18.32 16.51

Foot length 16.08 19.85 15.53 17.28 15.91

T4 lamellae eA of 18 20 21

Sex M F F M M

Diagnosis. A large-bodied gecko (SVL 116—128.83 — folds; 21-25 transverse rows of ventrals; 10—12

mm) restricted to the southern islands of the Nicobar

archipelago, characterized by: 14-17 supralabials; 12 or

13 infralabials; two elongate inner pair of postmentals

in broad medial contact with each other; two smaller,

separated outer pairs of postmentals; 13-15 precloacal

pores in males, no femoral pores; two internasals in

contact with each other; distinct ventrolateral dermal

Amphib. Reptile Conserv.

transverse rows of enlarged, rounded dorsal tubercles;

three enlarged post-cloacal spurs on each side of the

vent; 18—22 undivided subdigital lamellae under toe IV:

presence of five legible, light-colored, creamy white,

transverse bands in juveniles, subadults and adults have

a pale-white to creamy yellow venter.

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Chandramouli et al.

Fig. 9. Lateral, ventral, and dorsal profiles of the head, and precloacal region of the holotype of Gekko stoliczkai sp. nov.

Description of the holotype (ZSI/ANRC/T/6092). An

adult male, measuring 118.37 mm SVL, head fairly large

(HL:SVL 0.27), longer than broad (HL:HW 1.34); with

a blunt, rounded snout tip. Eyes fairly large (ED:HL

0.24) with a vertically elliptical pupil; eye slightly

smaller than the snout length (ED:ES 0.52); nostrils

situated closer to the snout tip than to the eyes (EN:ES

0.78). Trunk slightly shorter than one-half the length of

the body (AG:SVL 0.47). Overall habitus depressed.

Supralabials 14 on each side, infralabials 13 on each side;

two moderately enlarged postmentals in broad medial

contact, followed by two pairs of enlarged scales, that are

nearly as large as the post-mentals. Dorsum bearing 11

transverse rows of enlarged, rounded tubercles. Ventrals

imbricate, in 22 transverse rows. Two pairs of enlarged,

rounded cloacal spurs present at the base of the vent on

each side. Subcaudals horizontally elongate, the mid-

anterior scales not enlarged. Tail slightly shorter than

the body (SVL:TAL 1.12). Upper arm shorter than lower

arm (UAL:LAL 0.82); palm with enlarged, undivided

subdigital lamellae; the first one with an indistinct claw;

relative lengths of fingers IV>III>V>II>I. Thighs short

(FEL:SVL 0.17) and robust; with a few tubercles on

the dorsal surface. Tibia shorter than thighs (TBL:SVL

0.15); toes with entire, undivided subdigital lamellae, 21

on toe IV; relative lengths of toes IV>III>V>II>I. Fifteen

undivided series of precloacal pores; the pore-bearing

scales relatively smaller than those above. Fingers and

toes free, lacking membranous skin flaps.

Amphib. Reptile Conserv.

Coloration. In life, overall ground coloration dull-brown

with five or six indistinct pale-white transverse cross-

bars on the body. Tail regenerated and uniform brown.

Venter brown with small brown spots on each ventral

scale. Eyes with vertically elliptical pupils and a bluish

iris. In preservation, the dorsal coloration faded to a

near-uniform dull brown with a pale white venter. The

transverse bars barely visible and the bluish coloration

of the iris faded.

Variation. Measurements and scale counts of the

paratypes are given in Table 4. Females lack precloacal

pores and are nearly as large as males. Light-colored,

creamy white, transverse bands are more legible in

juveniles and subadults while the adults usually have

feeble dorsal bands. Ventral color ranges from pale-white

to creamy yellow.

Natural history. Nocturnal and found in a variety of

habitats ranging from evergreen forests, semi-evergreen

forests, and plantations to human habitations. Frequently

observed on walls of buildings, or on tree trunks of tall

trees at heights ranging from about 6 to 13 feet. Calls

comprise a series of repeatedly uttered, interrupted

rattling syllables of tuk... tuk...tuk...tuk advancing into

high frequency syllables of tuk-tuk-tuk-tuk (also see

Biswas 1984).

Comparisons. Gekko stolickzkai sp. nov. can be

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New Gekko species from Andaman and Nicobar Islands

91°30'0"E 92°30'0"E

10°30'0"'N 11°30'0"N 12°30'0"'N 13°30'0"N

9°30'0"N

93°30'0"E

Andaman

94°30'0"E 95°30'0"E

13°30'0"N

Islands

12°30'0"'N

10°30'0"'N 11°30'0"N

9°30'0"N

8°30'0"'N

7°30'0"N

0 35 350 100 km

91°30'0"E 92°30'0"E

Sombrero Channel----

Little & @

Great

Nicobar

93°30'0"E

Central Nicobars

8°30'0"'N

7°30'0"N

94°30'0"E 95°30'0"E

Fig. 10. Map of the Andaman and Nicobar Islands showing the distributions of Gekko verreauxi (red), Gekko nicobarensis (yellow),

and Gekko stoliczkai sp. nov. (black).

differentiated from G. smithii by having significantly

fewer numbers of internasals and ventrals and having

significantly higher numbers of infralabials and

supralabials (Tables 1-3; Figs. 1-2). It can be further

separated from G. smithii by its less gracile more stout

body features, in having a significantly narrower and

shorter head and snout, a significantly smaller orbit,

Amphib. Reptile Conserv.

and significantly shorter limbs and trunk. From G.

verreauxi, it 1s differentiated by the separation of nasal

and rostral scales (vs. in contact in G. verreauxi), bluish

iris (vs. greenish in G. verreauxi), and greater number of

supralabials (14—17 in G. stoliczkai sp. nov. vs. 12—14 in

G. verreauxi). From Gekko nicobarensis, G. stoliczkai sp.

nov. can easily be distinguished by the absence of skin-

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Chandramouli et al.

flaps along the sides of the body and tail, and extensive

webbing between fingers and toes (vs. present in G.

nicobarensis), and by the separation of nasal and rostral

scales (vs. in contact in G. nicobarensis). Additionally,

from other members of the Gekko gecko group, the new

species G. stoliczkai sp. nov. could be distinguished as

follows (only opposing suite of characters mentioned):

G. albofasciatus (16 precloacal pores and reddish olive

dorsal coloration), G. gecko (11-15 supralabials; gray to

bluish or brownish dorsal color with reddish spots), G.

nutaphandi (12—14 supralabials; 17—22 precloacal pores

in males; 15 subdigital lamellae under toe IV), G. reevesii

(10-14 supralabials; 13—20 precloacal pores; gray-green

to dark grey dorsal coloration), and G. siamensis (13-21

supralabials; 10—13 precloacal pores; grey-brown to dark

green dorsal coloration) fide Rosler et al. (2011).

Distribution. Recorded during the present study from

Great and Little Nicobar Islands. It has been reported

from other smaller islands such as Pigeon, Pilo Milo,

Menchal, and Kondul in the southern group of the

Nicobar Islands (Vijayakumar 2005) (Fig. 10).

Discussion

A fairly recent taxonomic review of the genus Gekko by

Rosler et al. (2011) defined six species groups based on

morphological and phylogenetic evidence and classified

the members that formed six sub-clades. As per their

scheme of classification, Gekko smithii and G. verreauxi

fall under the Gekko gekko group, among which, G.

smithii was shown to comprise a complex of several

populations within the Sundaland. The subsequent

classification of this group under the subgenus Gekko by

Wood et al. (2019) is in agreement with that of Rosler

et al. (2011). Although Das (1999) doubted that the

Nicobar population of G. smithii was potentially allied

to an undescribed member of this group from Sumatra

and specifically distinct from the nominate species,

Gekko smithii, no taxonomic assessments of this

population have been made until now, despite several

of the earlier field-based observations, collections, and

records (Biswas and Sanyal 1977; Vijayakumar 2005;

Harikrishan et al. 2014). The present study has filled

in this long-standing lacuna by providing taxonomic

clarity for the Nicobarese population. A report of the

‘presence’ of Gekko smithii from Camorta in the central

Nicobar Islands by Harikrishnan et al. (2014: 18) is

erroneous and it does not occur there (pers. obs.).

Additionally, this study provides new data on the

other poorly-known members of the genus Gekko

found in the Andaman and Nicobar Islands, especially

G. verreauxi, which is restricted to the Andaman

archipelago. Although earlier studies provided some

information on taxonomy and field observations (Ota

et al. 1991; Vesely 1999), this species has remained

poorly known until now. Mohan et al. (2020) studied

Amphib. Reptile Conserv.

intraspecific genetic variation within G. verreauxi across

the Andaman Islands and found it to be low, implying

a genetic uniformity across the island populations,

as expected (e.g., as in Phelsuma andamanensis and

Cyrtodactylus rubidus). The other species, Gekko

(Ptychozoon) nicobarensis was variously attributed

to Ptychozoon homalocephalum and Ptychozoon

kuhli historically (Stindachner 1867), until Das and

Vijayakumar (2009) identified this population to be

specifically distinct, thereby conferring it the name

Ptychozoon nicobarensis. They diagnosed the species

based on ‘dorsum with tan vertebral stripe, lacking

dark transverse bars’ (Das and Vijayakumar 2009:

10). Their description was based on a commendable

series of 20 specimens, comprising seven males and

13 females. However, our new material for this species

indicates that it is much more variable in morphology

and color pattern than previously ascertained, and

can have either a striped or banded dorsal pattern

(Fig. 3). The Andaman Islands are separated from the

Nicobar Islands by the Ten-Degree Channel, which

acts as an effective biogeographic barrier, limiting the

distribution of G. verreauxi. Likewise, the Sombrero

Channel running between the central and southern

groups of Nicobar Islands act as a barrier in limiting

the distributions of G. nicobarensis and G. stoliczkai

sp. nov. The Great Channel, running between Great

Nicobar and Sumatra, acts as a biogeographic barrier

which separates the distribution ranges of G. stoliczkai

sp. nov. (Southern Nicobar Islands) and the Sundaic

Island of Sumatra, occupied by Gekko smithii. Among

these, the conservation status has been assessed only

for ‘G. smithi? as Least Concern. Considering the

relatively wide geographic distribution and fairly high

abundance (pers. obs.), we recommend G. verreauxi

to be classified under the Least Concern category,

while the other two species, G. (P.) nicobarensis and

Gekko stoliczkai sp. nov., with their distribution ranges

restricted to a few relatively smaller islands, would fall

under the Endangered category, owing to their much

narrower geographic distributions.

Acknowledgements.—We thank the Department of

Environment and Forests, Andaman and Nicobar Islands

for permission (permit numbers: CWLW/WL/134/

(J)/Folder/417 and CWLW/WL/134 (L)/ 60) allowing

SRC to conduct this study and for the infrastructure

provided. SRC is thankful to the Mohamed bin Zayed

Species Conservation Fund for grants (#14058387 and

#160514249) which partly facilitated this study. We

thank the faculty of the Department of Ecology and

Environmental Sciences and the Department of Ocean

Studies and Marine Biology, Pondicherry University, for

the lab space and support extended. CS and GG thank

Kailash Chandra, Director, Zoological Survey of India,

Kolkata, for his support, cooperation, and encouragement

during the survey.

June 2021 | Volume 15 | Number 1 | e276

New Gekko species from Andaman and Nicobar Islands

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Chandramouli et al.

S.R. Chandramouli obtained his Doctoral Degree in Ecology and Environmental Sciences from

Pondicherry University, India. His work focuses on the systematics, taxonomy, ecology, and

biogeography of the squamate reptiles and amphibians of peninsular India and the Andaman and

Nicobar Islands, and has resulted in the discovery of two new species of amphibians, five new

lizards and a new snake. He serves as a member of several committees for conservation within the

IUCN.

G. Gokulakrishnan works as Research Associate at the Zoological Survey of India, Port Blair,

India. He has received his Master’s Degree in Wildlife Biology from Bharathidasan University,

India, and is pursuing his Doctorate Degree on studies of the avifauna of important bird areas in the

Andaman and Nicobar Islands. His work focuses on the birds, reptiles, and amphibians of Andaman

and Nicobar Islands, and he has reported more than 40 new records of birds for the Andaman and

Nicobar Islands.

Chandrakasan Sivaperuman works as Scientist-E and Officer-in-Charge at the Zoological Survey

of India, Port Blair, India. He has been extensively involved in field surveys in different ecosystems

of the country, including Kole wetlands of Kerala, Southern Western Ghats, Eastern Ghats, Great

Indian Deserts, Andaman and Nicobar Islands, and has studied birds and mammals in Antarctica.

Chandrakasan has reported more than 50 new records for India from various faunal groups,

especially birds, mammals, moths, wasps, and odonates.

L. Lee Grismer studies phylogeny, taxonomy, biogeography, and character evolution in amphibians

and reptiles from Southeast Asia. Lee has been working throughout the region for over 25 years, and

he has contributed significantly towards studies on the systematics of the Oriental gekkonid genus

Cyrtodactylus.

Appendix 1. Comparative material of Gekko smithii examined.

Peninsular Malaysia: LSUHC 3891, LSUHC 3990, LSUHC 4681, LSUHC 4694, LSUHC 4697, LSUHC 5059,

LSUHC 5060, LSUHC 5061, LSUHC 5062, LSUHC 5063, LSUHC 5151, LSUHC 5152, LSUHC 5390, LSUHC

5399, LSUHC 5849, LSUHC 6260, LSUHC 6265, LSUHC 6277, LSUHC 6278, LSUHC 6282, LSUHC 6283,

LSUHC 6284, LSUHC 6542, LSUHC 6564, LSUHC 6606, LSUHC 6890, LSUHC 7024, LSUHC 7025, LSUHC

7026, LSUHC 7251, LSUHC 7257, LSUHC 7263, LSUHC 7264, LSUHC 7299, LSUHC 7648, LSUHC 7649,

LSUHC 7650, LSUHC 7651, LSUHC 7694, LSUHC 7702, LSUHC 9153, LSUHC 9154, LSUHC 9155, LSUHC

9156, LSUHC 9157, LSUHC 9158, LSUHC 9626, LSUHC 9959, LSUHC 10585, LSUHC 10596, LSUHC 11976,

LSUHC 11977, LSUHC 12028, LSUHC 13476, LSUHC 13624, LSUHC 13625, LSUHC 13626, LSUHC 13627,

LSUHC 15041, LSUHC 15042, LSUHC 15052, LSUHC 15053, LSUHC 185107, LSUHC 185112, LSUHC 185113,

LSUHC 185119, LSUHC 185126, LSUHC 185135, LSUHC 185136, LSUHC 185139, LSUHC 185141, LSUHC

185142, LSUHC 185144. FMNH 185145, FMNH 185114, FMNH 185120.

Thailand: THMNH 1841, THMNH 1844, THMNH 3247, THMNH 11432, THMNH 13910.

Sumatra: FMNH 209498503, USNM 29396, USNM 197804; MVZ 2112225; ZMA 17947A-B, 17952.

Borneo: FMNH 128893—95, FMNH 129492, FMNH 129495—96, FMNH 131520—-22, FMNH 230176, 246237—28,

248986; ZRC 2.5302, ZRC 2.5691, ZRC 2.5704, 2.1488.

Amphib. Reptile Conserv. 125 June 2021 | Volume 15 | Number 1 | e276