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smithsonian contributions to zoology • number 635

Variation, Systematics, and 

Relationships of the 

Leptodactylus bolivianus

Complex (Amphibia: 

Anura: Leptodactylidae) 

W. Ronald Heyer and Rafael O. de Sá
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smithsonian contributions to zoology • number 635

Variation, Systematics, and

Relationships of the

Leptodactylus bolivianus

Complex (Amphibia: 

Anura: Leptodactylidae)

W. Ronald Heyer and Rafael O. de Sá

washington d.c.

2011
ABSTRACT

Heyer, W. Ronald, and Rafael O. de Sá. Variation, Systematics, and Relationships of the Leptodactylus bolivianus Complex (Amphibia: Anura: Leptodactylidae). Smithsonian Contributions to 

Zoology, number 635, viii + 58 pages, 21 figures, 20 tables, 2011.—The Leptodactylus bolivianus

complex has been considered to consist of one or two species, L. bolivianus alone or L. bolivianus

and L. insularum. Detailed morphological analyses were undertaken to evaluate variation in the 

complex, which ranges from Costa Rica through Panama, across northern South America in the 

river valleys draining to the Caribbean, and throughout much of the Amazon basin with southern 

limits in Bolivia. Members of the complex also occur on several islands off Nicaragua, Panama, 

and Colombia. Analyses of morphological and advertisement call data indicate that there are 

either two or three species comprising the complex. Analysis of molecular data strongly supports 

recognition of three species, one of which is described as a new species, Leptodactylus guianensis. 

The three species comprising the L. bolivianus clade are most closely related to the L. ocellatus

clade within the genus Leptodactylus.

Cover photos, from left to right: Leptodactylus insularum (Figure 17, courtesy of R. W. McDiarmid), L. guianensis (Figure 15, courtesy of M. S. Hoogmoed), and L. bolivianus (Figure 

12, courtesy of R. B. Cocroft).

Published by SMITHSONIAN INSTITUTION SCHOLARLY PRESS

P.O. Box 37012, MRC 957

Washington, D.C. 20013-7012

www.scholarlypress.si.edu

Text and images in this publication may be protected by copyright and other restrictions or owned by individuals and entities other than, and in addition to, the Smithsonian Institution. Fair use of copyrighted material includes the use of protected materials for personal, educational, or noncommercial purposes. Users must 

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Library of Congress Cataloging-in-Publication Data

Heyer, W. Ronald.

Variation, systematics, and relationships of the Leptodactylus bolivianus complex (Amphibia: Anura: 

Leptodactylidae) / W. Ronald Heyer and Rafael O. de Sá.

 p. cm. — (Smithsonian contributions to zoology ; number 635)

Includes bibliographical references.

1. Leptodactylus bolivianus—Classification. 2. Leptodactylus bolivianus—Variation. I. De Sá, Rafael O.

II. Title. 

QL668.E257H49 2011

597.8’75—dc22

2011004864

ISSN: 0081-0282 (print); 1943-6696 (online)

The paper used in this publication meets the minimum requirements of the American National Standard 

for Permanence of Paper for Printed Library Materials Z39.48–1992.
Contents

LIST OF FIGURES v

LIST OF TABLES vii

INTRODUCTION 1

METHODS AND MATERIALS 1

Juvenile and Adult Morphological Data 2

Larval Morphology 3

Advertisement Calls 4

Molecular Sequence Data 5

Measurement DATA 5

INTRASAMPLE VARIATION 6

Dorsal Pattern 8

Subocular Spot 9

Lip Stripe 9

Belly Pattern 9

Thigh Pattern 9

Shank Pattern 9

Dorsolateral Folds and Lateral Folds 9

Male Secondary Sexual Characters 9

Measurement Data 9

INTERSAMPLE VARIATION 12

Patterns and Folds 12

 Dorsal Pattern 12

 Subocular Spot 14

 Lip Stripe 17

 Belly Pattern 19

 Thigh Pattern 19

 Shank Pattern 20

 Dorsolateral Folds 21

 Lateral Folds 21
i v • smithsonian contributions to zoology

Male Secondary Sexual Characters 21

Measurement Data 22

GEOGRAPHIC VARIATION AMONG THE SAMPLES 23

Adult Form Morphological Characters 23

Larval Morphology 24

Advertisement Calls 25

 Costa Rica Recording 25

 Panama Recording 25

 Peru Recording 25

MOLECULAR RELATIONSHIPS 26

INTERSPECIFIC VARIATION 27

Measurement Data 28

Morphological Data 29

 Leptodactylus bolivianus and L. insularum Comparisons 30

 Leptodactylus bolivianus: New Species Comparisons 30

 Leptodactylus insularum: New Species Comparisons 30

SPECIES ACCOUNTS 30

Leptodactylus bolivianus Boulenger, 1898 31

Leptodactylus guianensis, new species 35

Leptodactylus insularum Barbour, 1906 38

ACKNOWLEDGMENTS 45

APPENDIX 1: OBSERVER ERROR ASSESSMENT 

FOR MORPHOLOGICAL DATA 47

APPENDIX 2: molecular sample LOCALITY DATA 55

REFERENCES 57
Figures

1. Locations of specimens used for intrasample analyses 7

2. Locations of 20 regional samples 17

3. Variation in posterior thigh patterns among 20 regional samples 20

4. Female discriminant function analysis results of 20 geographic 

samples 23

5. Levels of differentiation between geographically adjacent samples 23

6. Advertisement call of Leptodactylus from Costa Rica 26

7. Advertisement call of Leptodactylus from Panama 27

8. Advertisement call of Leptodactylus from Peru 28

9. Maximum likelihood cladogram of the Leptodactylus ocellatus 

species group 29

10. Plot of first two canonical vectors for male measurement data 29

11. Plot of first two canonical vectors for female measurement data 30

12. Leptodactylus bolivianus from Tambopata, Peru 32

13. Male thumb spines 32

14. Distributions of Leptodactylus bolivianus, L. guianensis, 

and L. insularum 34

15. Leptodactylus guianensis from northern Pará, Brazil 35

16. Lateral view of head, dorsal and ventral views of Leptodactylus

guianensis, new species 36

17. Leptodactylus insularum from San Andrés, Colombia 38

18. Dorsal and ventral views of the lectotype of Leptodactylus 

insularum 39

19. Lateral view of Leptodactylus insularum tadpole 40

20. Oral disk of Leptodactylus insularum tadpole 40

Appendix 1 figure

A1.1. Example of design pattern 50

Tables

1. Statistics to determine observer error 6

2. Dorsal pattern state occurrences within Lower Río Chucunaque 

sample 8

3. Dorsal pattern state occurrences by age and sex within Lower 

Río Chucunaque sample 8

4. Chi-square occurrence values for character states of the dorsal 

pattern within Lower Río Chucunaque sample 8

5. Age and sex intrasample analyses results for dorsal pattern 8

6. Dorsal pattern, suborbital spot, lip pattern, and belly pattern 

state occurrences 10

7. Age and sex intrasample analyses results for subocular spot 11

8. Age and sex intrasample analyses results for lip stripe pattern 11

9. Age and sex intrasample analyses results for belly pattern 12

10. Age and sex intrasample analyses results for thigh pattern 12

11. Thigh pattern, shank pattern, dorsolateral fold, and lateral 

fold occurrences 13

12. Age and sex intrasample analyses results for dorsolateral folds 14

13. Age and sex intrasample analyses results for lateral folds 14

14. Male secondary sexual character state occurrences 15

15. Intrasample variation in measurement data 16

16. Intersample comparisons for dorsal pattern data 18

17. Number of males exhibiting secondary sexual character states 22

18. Quantitative larval data 24

19. Advertisement call data 25

20. Morphological character states for species pairs of the 

Leptodactylus bolivianus complex 31

Variation, Systematics, and Relationships 

of the Leptodactylus bolivianus Complex 

(Amphibia: Anura: Leptodactylidae)

INTRODUCTION

A cluster of morphologically similar frogs of the genus Leptodactylus having a pair of distinct dorsolateral folds on the dorsum and well-developed lateral fringes on the toes has never been systematically evaluated by examining 

materials from throughout its geographic range. The species involved are herein 

referred to as members of the Leptodactylus bolivianus complex. There have 

been three names proposed for members of this complex: Leptodactylus bolivianus Boulenger, 1898; Leptodactylus insularum Barbour, 1906; and Leptodactylus romani Melin, 1941. The collective range for the L. bolivianus complex is 

from Costa Rica southward through Panama, extending across northern South 

America (east of the Andes) in the river valleys draining to the Caribbean, and 

throughout much of the Amazon basin with southernmost limits in the Bolivian 

departments of La Paz, Cochabamba, and Santa Cruz.

We analyze variation in this complex of frogs throughout its geographic 

range to understand inherent patterns of differentiation and to interpret those 

patterns in terms of species-level recognition, distributions, and relationships.

METHODS AND MATERIALS

We follow Fabrezi and Alberch (1996) in numbering the fingers II–V (considering finger I as lost in frogs). Herein, we use the name Leptodactylus latrans

following the recent elucidation of the nomenclature of the widely spread taxon 

previously referred to as L. ocellatus (Lavilla et al., 2010).

The primary materials examined for this study are juvenile and adult 

museum specimens; only this data set provides extensive locality records for 

the members of this complex. The juvenile and adult morphological data are 

W. Ronald Heyer, Division of Amphibians and 

Reptiles, National Museum of Natural History, Smithsonian Institution, MRC 162, P.O. 

Box 37012, Washington, D.C. 20013-7012, 

USA. Rafael O. de Sá, Department of Biology, 

University of Richmond, Richmond, Virginia 

23173. Correspondence: heyerr@si.edu. Manuscript received 28 June 2010; accepted 23 September 2010.
2 • smithsonian contributions to zoology

complemented by available data on larval morphology, 

advertisement calls, and molecular sequences.

The names of the institutions and collections from 

which specimen data were obtained are abbreviated here 

(after Sabaj Pérez, 2010) for subsequent mention in the 

text.

AMNH American Museum of Natural History

CM Carnegie Museum of Natural History

FMNH Field Museum of Natural History

ICNMHN Instituto de Ciencias Naturales, Museo 

de Historia Natural, Universidad Nacional de Colombia

IND Instituto Nacional de Recursos Naturales 

Renovables y del Ambiente, Bogota

INPA Instituto Nacional de Pesquisas da Amazonia, Brazil

KU University of Kansas Natural History 

Museum

LSUMZ Louisiana Museum of Natural History

MCZ Museum of Comparative Zoology, Harvard University

MNCN Museo Nacional de Ciencias Naturales, 

Madrid

MNRJ Museu Nacional, Universidade Federal 

do Rio de Janeiro

MPEG Museu Paraense “Emilio Goeldi,” Belém, 

Brazil

MPUJ Museo de Pontificia Universidade de Javeriana, Bogotá

MZUSP Museo de Zoologia da Universidade de 

São Paulo, São Paulo

NRM Naturhistoriska Riksmuseet, Department 

of Vertebrate Zoology, Stockholm

TCWC Texas Cooperative Wildlife Collection, 

Texas A&M University, College Station, 

Texas

TNHC Texas Natural History Collections, Texas 

Natural Science Center, Texas Memorial 

Museum, University of Texas at Austin

UMMZ University of Michigan Museum of Zoology, Ann Arbor, Michigan

USNM National Museum of Natural History, 

Smithsonian Institution, Washington, D.C.

UTA University of Texas at Arlington, Department of Biology, Texas

The following collections are not included in Sabaj 

Pérez (2010).

AL Adolfo Lutz collection housed at MNRJ

ULABG Universidad de Los Andes, Laboratorio 

de Biogeografía, Mérida, Venezuela

The following are field tag abbreviations, not yet incorporated into collections.

AMU Abraham Mijares

ELM Enrique La Marca

JC Jay Cole

RdS Rafael de Sá

UC Ulisses Caramaschi

USNM-FS National Museum of Natural History, 

Smithsonian Institution, Washington, D.C.

Juvenile and Adult Morphological Data

The data involved were taken and analyzed by the first 

author (WRH) for consistency (see Hayek et al., 2001). 

Because of the moderately large size of adult specimens 

of this complex and new restrictions on transportation of 

alcohol-stored specimens, it was necessary to take data on 

the specimens in the collections involved rather than borrowing the specimens. Thus only the USNM specimens, 

plus a few other specimens on loan to WRH, were available for reexamination during this study.

Specimen data from the following collections were 

taken in situ: AMNH, FMNH, ICNMHN, IND, KU, 

MCZ, MNRJ, Museo Universidad Javariana–Bogota, 

MZUSP, UMMZ, and USNM.

Geographic samples are analyzed to evaluate the nature of morphological variation. The samples are indicated 

by italics and leading capital letters. 

In 2005 an evaluation of the assembled locality data 

was undertaken to determine if there were any obvious 

geographic gaps for which specimens should be sought 

in other collections. There was only one obvious gap—

no data were available from the Brazilian State of Pará, 

even though members of the complex occur in extensive 

portions of Amazonia. The MZUSP and USNM collections contain rather extensive amphibian collections from 

Pará, suggesting that members of the complex really do 

not occur in Pará. The institution housing the most herpetological specimens from Pará is the MPEG. Dr. Marinus Hoogmoed reported that there were no specimens of 

the L. bolivianus complex catalogued in the MPEG collection (M. Hoogmoed, personal communication [e-mail] 

to WRH, 30 March 2005). At the VI World Congress of 

Herpetology in July 2008, Dr. Hoogmoed (pers. comm.) 

informed WRH that he had reexamined frogs from the 

state of Pará, near the state of Amapá and found them 

to be members of the Leptodactylus bolivianus complex. 
number 635 • 3

Subsequently (e-mail message of 20 December 2008 to 

WRH) Dr. Hoogmoed wrote, “During the last fieldwork 

again 4 specimens of Leptodactylus bolivianus [new species as recognized in this paper]. Thus the species is well 

distributed in Para north of the Amazon.” 

Specimens from the collections indicated above should 

be adequate to evaluate patterns of morphological variation throughout the distribution of the species complex.

After examination of materials from the AMNH, 

ICNMHN, IND, MCZ, MNRJ, MZUSP, and USNM, it 

seemed that data taken to that point on surface textures 

of the dorsal shank, outer tibia, and sole of foot did not 

demonstrate meaningful interpopulational variation. Before visiting further collections, an analysis was made (see 

Appendix 1) that confirmed the uninformative value of 

those characters, and no further data were taken for these 

variables. 

The variables recorded for the entire data set for analysis of variation organized by category are as follows: 

(1) Sex. Adult males are defined as having vocal slits. 

Juvenile males lack vocal slits. Gonads of individuals near 

adult size were examined to determine whether they were 

juvenile males or females. Adult females are defined as having at least curly oviducts. Juvenile females have straight 

oviducts. Most small specimens were not dissected to examine for gonads and are classified as juveniles. 

(2) Belly, dorsal, lip, posterior thigh, and dorsal shank 

patterns. Outline drawings were filled in for each of these 

characters, and these drawings served as pattern standards; additional standards were created as deemed appropriate. For data-recording purposes, each pattern was 

given a unique letter or combination of a letter and number. Specimens that had patterns intermediate between 

two standards where creation of a new standard was considered inappropriate were recorded as having both patterns separated by a slash. These slashed pattern notations 

are treated as separate states. Belly patterns were based 

on standards for the Leptodactylus pentadactylus species 

group (Heyer, 2005) as a starting point to which standards 

were added as needed. The dorsal, lip stripe, posterior 

thigh, and dorsal shank patterns used were initially created on the basis of small samples of specimens of both 

the L. bolivianus and L. latrans clusters with additional 

standards added in this study. The first two collections examined were MZUSP and USNM during which the majority of new standards were created. Subsequently, any new 

standard created was dated to know when in the study it 

was available for comparison.

(3) Dorsolateral and lateral folds. Combinations of 

letters, numbers, and parentheses were used in data coding 

to express the completeness of the pair of dorsolateral 

folds extending from the supratympanic fold to the groin 

along the junction of the dorsal and flank regions and the 

pair of lateral folds extending from the supratympanic 

fold to the groin in the midflank region. 

(4) Male secondary characteristics. Combinations of 

numbers, letters, pluses, minuses, and parentheses were 

used to code variation in male arm hypertrophy, tympanic 

tubercles, jaw tubercles, chest tubercles, and thumb spines. 

(5) Measurements. Measurement data were taken 

with calipers either with the unaided eye or under a dissecting microscope for snout–vent length (SVL), head 

length, head width, shank length, and foot length following Heyer (2005). Measurements on MZUSP specimens 

were recorded to the nearest 0.5 mm; all others were recorded to the nearest 0.1 mm.

Observer error increases when materials are not 

available for reexamination. Therefore this problem was 

specifically addressed before the data were analyzed for 

geographic variation. The methods used to determine observer error are described in Appendix 1.

Larval Morphology

Terminology and methods follow McDiarmid and 

Altig (1999) including use of Gosner developmental 

stages. In addition to the examined USNM larval collection for samples of this complex, AMNH, CM, FMNH, 

ICNMHN, KU, MCZ, and UMMZ were contacted for 

larvae identified as either L. bolivianus or L. insularum in 

their collections.

There are very few larval samples from the above collections. Only KU and USNM had samples identified as 

belonging to the L. bolivianus-insularum complex. The 

KU samples were borrowed and examined by WRH.

The KU larval specimens from two localities have 

been described in the literature (Duellman, 1997, 2005). 

Upon examination of these specimens, WRH determined 

that the published descriptions were based on misidentified specimens. Duellman (1997) described KU 167784 

and 167785 from km 104 on the road from El Dorado to 

Santa Elena de Uairén, Bolívar, Venezuela, as L. bolivianus. 

No juvenile or adult KU specimens of the L. bolivianusinsularum complex were collected at this locality. The tadpoles are in fair condition, very soft, with all teeth missing. 

Duellman (1997) indicated that seven individuals at Gosner stage 25 had body lengths of 15.5–18.4 mm. Data 

were taken on five larvae from each sample that visually 

appeared to cover the size ranges for each sample. Tadpole stages ranged from 26 to 35 with body sizes ranging 
4 • smithsonian contributions to zoology

from 14.0 to 18.1 mm. Duellman (1997) did not report a 

tooth row formula for the larvae. Two tadpoles were examined using crystal violet stain to differentiate the tooth 

row ridges that bear the teeth. The anterior ridges were 

unclear in one individual, but the other individual has a 

3(3)/3(1) formula. Both tadpoles have the same posterior 

tooth row condition, with a short P3 row, which is unusual for Leptodactylus larvae but does occur in a few 

members of the L. pentadactylus species cluster, such as in 

some individuals of L. knudseni (Heyer, 2005). The vent 

appears to be median, although the condition of preservation makes the evaluation difficult. Given the 3(3)/3(1) 

tooth row formula, the larvae most likely do not belong to 

the genus Leptodactylus. Duellman (2005) described and 

illustrated larvae of L. bolivianus from Cusco Amazonico, 

Madre de Dios, Peru. The specimen illustrated in Duellman (2005:177, fig. 13.10A) was placed in a separate 

container and identified as the illustrated specimen. There 

is one discrepancy between the specimen, Duellman’s description, and the illustration. Duellman (2005:279) described a specimen at stage 25. The illustration shows a 

limb bud with early differentiated digits, perhaps at stage 

35 or 36, but not of normal shape for digits at those stages 

of development. Duellman (2005) reported the tooth row 

formula as 2/3(1). Reexamination of the illustrated specimen indicates a formula of 1/3(1), with a very short P3 

row. Examination of several specimens from KU 205810 

indicates the variable formula 1/2–3(1), with P3 always 

very short when present. A single tadpole was examined using crystal violet stain. A broadly separated pair 

of ridges with two lateral extensions is present where an 

A2 row would be expected. As indicated, none of these 

ridges bear teeth in the several specimens examined from 

lot KU 205810. The tadpoles identified as L. bolivianus

(Duellman, 2005) do not belong to the genus Leptodactylus; most likely, they are unidentified larvae of leiuperid, 

leptodactylid, or dendrobatid frogs.

Advertisement Calls

The same institutions that were contacted for larval 

holdings were contacted for holdings of any recordings of 

advertisement calls. Two recordings were available at the 

Texas Natural History Collection. Both recordings were 

from the canal zone region of Panama, one of poor quality. The better recording was not requested, as there was 

another recording available for the same area. In addition, 

César Barrio and Enrique La Marca reported that they did 

not know of any recordings of the L. bolivianus-insularum

complex from Venezuela.

There are three commercially available CDs containing purported recordings of specimens of the L. bolivianusinsularum complex. 

Márquez et al. (2002) contains two brief recordings of advertisement calls identified as L. bolivianus. 

The recordings were obtained near a pond where several 

males were calling, and no particular voucher was collected (Ignacio De la Riva, pers. comm. [e-mail], 6 July 

2006). These calls from Bolivia are very different from 

those recorded from Tambopata, Peru, for which there is 

a museum voucher specimen corresponding to L. bolivianus (USNM 343347). Either the Bolivian recording species identification is in error or the Bolivian and Peruvian 

frogs associated with the calls represent distinct species. 

The Bolivian recordings have considerable sound in the 

same broadcast channel as the call of interest and cannot 

be analyzed in much detail. On the basis of the available 

information, these Bolivian CD recordings are considered 

to not belong to a member of the L. bolivianus-insularum

complex.

The Cusco Amazónico recording KU cassette 104/3 

published and figured by Duellman (2005:279) for L. bolivianus was requested for analysis. KU cassette 104/3 

was mistakenly discarded before it was incorporated 

into the KU tape archives (W. E. Duellman, pers. comm., 

e-mail message of 27 June 2006). The description provided (Duellman, 2005:279) could represent either L. bolivianus or more probably (on the basis of the statement, 

“The call consists of a continuous bubbling sound”), 

L. leptodactyloides.

Two Costa Rican recordings were borrowed for 

analysis: KU 801 and KU 802. The KU 801 recording has 

minor tape speed problems. The KU 802 recording has 

major tape speed problems and was not analyzed.

Three recordings were analyzed with Raven 1.2 software (Charif et al., 2004); terminology follows Heyer et 

al. (1990). Data for beginning, highest, and ending frequencies were taken from audiospectrogram displays. 

Dominant frequencies were obtained from spectrum analyses of entire calls. Call durations were taken from waveform displays. All calls were filtered around the broadcast 

frequencies for analysis. Default settings were used for audiospectrograms (window: Type Hann, size 512 samples, 

3 dB filter bandwidth 124 Hz; time/grid: overlap 50%, 

hop size 256 samples; frequency grid: DFT size 512 samples, grid spacing 86.1 Hz; averaging: 1 spectra) and spectrums (window: size 256 samples, 3 dB filter bandwidth 

248 Hz; time grid: overlap 50%, hop size 128 samples; 

frequency grid: DFT size 256 samples, grid spacing 172 

Hz; averaging: 1 spectra).
number 635 • 5

Molecular Sequence Data

Total genomic DNA was extracted using DNAeasy 

kit (USB) from liver or thigh muscle that was preserved 

in ethanol 95%. A segment of about 850 base pairs (bp) 

from the 16S ribosomal RNA (rRNA) gene was polymerase chain reaction (PCR) amplified (Palumbi, 1996) 

using an MJ Research PTC-200 thermocycler. Doublestranded PCR amplifications were performed using Green 

Taq Master Mix (Promega). 

Amplification primers used were 16S Ar 

5-CGCCTGTTTACCAAAAACAT-3 and 16S Br 

5-CTCCGGTCTGAACTCAGATCACGTAG-3 under 

the following thermal conditions: initial denaturation at 

94°C for 2 min followed by 34 cycles of 94°C for 1 min, 

58°C for 1 min, and 72°C for 1 min 30 s. Amplified segments were purified using Exo-Sap (USB) by heating 

samples at 80°C for 15 min. Purified products were cycle 

sequenced with the dideoxy chain termination method 

using the Sequi-Therm Excel II DNA sequencing kit (Epicentre Technologies). Infrared-labeled sequencing primers 

(same as amplification primers) were used in sequencing reactions (LiCor Biotechnology) under the following 

thermal conditions: initial denaturation at 95°C for 2 min 

30 s, followed by 30 cycles of 95°C for 30 s, 58°C for 

30 s, and 70°C for 30 s. Sequencing products were run in 

6% acrylamide, 44 cm in length, gels using a Licor DNA 

4300 automatic sequencer. Sequencing reactions were 

single stranded; double-stranded PCR fragments were 

sequenced in both directions. Sequences included in the 

analyses represent a consensus of both DNA strands; GenBank accession numbers for the sequence data are: HQ 

232831–232846, HQ259119, HQ259120, AY947857, 

AY911285, AY669856, AY943235, EF632048, EF632053, 

and EF632055

Complementary sequence strands for each sample 

were first aligned and, using the chromatographs created by BaseImagr software (LicorBiotechnology), were 

inspected visually for mismatches of aligned positions to 

confirm or manually correct the automatic reading. Consensus sequences were aligned with ClustalX (Thompson 

et al., 1997) using the multiple-alignment option. Phylogenetic analyses of sequence data were run using maximum 

parsimony (MP) and maximum likelihood (ML) criteria as 

implemented in PAUP* 4b6 (Swofford, 2002). Character 

states were treated as unordered, and gaps were considered as a fifth character (MP analysis); heuristic searches 

were performed with 100 random additions of sequences 

and tree bisection-reconnection (TBR) branch swapping. 

Strict consensus trees were calculated when several equally 

parsimonious trees resulted from the MP searches. Maximum likelihood analyses used the general time-reversible 

model with empirical base frequencies, gamma distribution of across-site rate variation, and an estimated proportion of invariable sites (GTR + Γ + I; Swofford et al., 1996) 

recommended by ModelTest 3.04 (Posada and Crandall, 

1998). Clade support of inferred trees was assessed by 

nonparametric bootstrapping (Felsenstein, 1985) on the 

basis of 100 pseudoreplicates.

Seventeen available samples from the L. bolivianusinsularum complex (additional data provided in Appendix 

2) in addition to a sample of L. latrans and of L. chaquensis were considered as the ingroup taxa (traditionally 

all included in the L. latrans species group). To include 

variation found in the genus Leptodactylus (sensu stricto), 

the following taxa were used as outgroups: L. podicipinus, L. wagneri (melanonotus species group), L. mystacinus, L. gracilis (fuscus species group), L. paraensis, and 

L. knudseni (pentadactylus species group).

Measurement Data

The protocol recommended by Hayek et al. (2001) of 

measuring a single individual male and female 20 times 

was followed. For this study, two sets of specimens were 

remeasured. The first set represents well-preserved specimens USNM 200352 (female) and USNM 200353 (male). 

Twenty-one data sheets were prepared to record data (to 

allow for at least one unambiguous error to be discarded 

and still have 20 data observations). Only a single sheet 

was filled out on any given day. These data were taken 

on the same days other data were being taken on USNM 

specimens. The sheets were filled in such that male data 

were recorded first on alternate sessions and the sheets 

were filled out alternately at either the beginning of the 

data taking session or the end of the session. Data were 

taken between 19 August and 28 October 2004. The second set represents stiff specimens (presumably yielding 

larger measurement errors): USNM 145708 (male) and 

USNM 145711 (female). Twenty-one data sheets were 

prepared to record data. A single sheet was filled out on 

any given day. Male data were recorded first on alternate 

recording sessions. Data were taken between 21 September and 21 October 2005.

Surprisingly, the well-preserved specimens were not 

notably measured more accurately than the stiff specimens 

(Table 1). For example, half of the well-preserved specimen 

variables had the highest coefficient of variation values. 

As it makes little or no difference which male and female 
6 • smithsonian contributions to zoology

data set is used in subsequent analyses, the male data for 

USNM 145708 and female data for USNM 145711 with 

the greatest magnitude of differences in coefficient of variation scores (comparing within sex) are selected for use in 

subsequent analyses.

The measurement data have such a low power of resolution to separate geographic and species samples that 

inclusion of the multiple measurement data from single individuals would be superfluous and were not incorporated 

in any discriminant function analyses.

INTRASAMPLE VARIATION

Variation of characters within samples is necessary as 

background information to understanding species variation. Also, for the nonsexually dimorphic morphological 

features, the intrasample data are evaluated to determine 

whether the characters demonstrate age or sex variation. 

If the characters vary by either age or sex, the characters 

need to be analyzed separately by those categories. The 

available data for this study do not have large sample sizes 

for either intra- or intersample analyses. Given that fact, 

there is an advantage to combining data by either age or 

sex in order to increase sample sizes for analysis, when it 

is appropriate to do so.

Ideally, the intrasample analyses should be based on 

specimens from single localities from throughout the geographic distribution of the complex. However, there are 

not adequate data from some major geographic areas. 

Some intrasample data sets therefore comprise data 

pooled from nearby localities rather than single localities. 

The following samples are used for intrasample variation 

analyses (Figure 1): (1) Costa Rica/Panama Border region 

(8 localities, n = 72); (2) Panama Canal region (23 localities, n = 46); (3) Lower Río Chucunaque, Panama region 

(12 localities, n = 58); (4) Ilha do Maracá, Brazil (1 locality, n = 36); (5) Boca do Acre, Brazil (1 locality, n = 36); 

and (6) Cuzco Amazónico, Peru (1 locality, n = 32).

The basic approach to analyzing the data in terms of 

whether the data are combined or analyzed separately by 

age or sex is to be conservative in combining data. That is, 

if the data show any reasonable evidence for demonstrating variation by age or sex, that is sufficient for intersample analyses to be analyzed separately by the appropriate 

categories.

The dorsal pattern data from the Lower Río Chucunaque, Panama are used to explain how the data were analyzed. The raw data for this character are shown in Table 

2, where five categories are used to summarize occurrence 

data for each character state: juvenile, juvenile male, juvenile female, male, and female. As not all nonadult specimens were dissected to verify sex, the categories juvenile 

versus juvenile male and juvenile female are not entirely discrete, but overall they represent a size difference with specimens categorized as juveniles being in most cases smaller 

than those categorized as juvenile males or juvenile females. 

The data from Table 2 were combined by either age or sex 

(Table 3). The observer error analysis for dorsal pattern indicated that there had to be a greater than 33% difference 

TABLE 1. Summary statistics for specimens remeasured 21 times to determine observer error. Definitions: Min, minimum measurement; 

Max, maximum measurement; CV, coefficient of variation; SVL, snout–vent length; HL, head length; HW, head width; EN, eye-nostril 

distance; TD, tympanum diameter.

Well-preserved specimens Stiffly preserved specimens

♂ USNM 200353 ♀ USNM 200352 ♂ USNM 145708 ♀ USNM 145711

Specimen variable Min Max CV Min Max CV Min Max CV Min Max CV

SVL 87.8 88.9 0.004 87.4 89.3 0.007 87.4 88.5 0.003 75.8 76.8 0.003

HL 30.8 33.3 0.019 31.2 32.2 0.011 30.3 32.3 0.016 26.8 29.3 0.026

HW 31.0 32.1 0.010 29.1 30.9 0.015 30.0 31.1 0.008 26.2 27.5 0.013

EN 8.3 8.7 0.013 8.0 8.6 0.018 8.1 8.4 0.011 7.7 8.0 0.011

TD 6.2 6.6 0.014 6.0 6.3 0.018 5.8 6.3 0.020 5.1 5.6 0.026

Thigh 39.0 39.8 0.005 40.5 42.5 0.013 37.6 38.5 0.007 34.0 35.3 0.009

Shank 39.8 40.8 0.006 43.3 43.8 0.003 42.2 43.3 0.008 39.3 40.2 0.007

Foot 42.2 43.6 0.009 44.4 46.2 0.010 42.5 45.3 0.014 40.4 42.1 0.012
number 635 • 7

in state distributions to be meaningful. The comparison of 

juvenile with adult data are determined by subtracting the 

smaller percent value from the larger percent value for the 

States A, B, and C, as indicted in the formula ((37 − 5) + 

(13 − 0) + (82 − 63)) = 64%. Thus the dorsal pattern character state distribution for the juvenile/adult categorization 

exceeds the 33% threshold for observer error. For cases 

such as this, chi-square analyses were performed to determine whether the differences were statistically significant. 

In this case, only two states had expected values ≥5 (Table 

4); accordingly, the chi-square analysis was performed only 

on the two character states with expected values ≥5. These 

analyses are thus conservative and need to be evaluated 

with that in mind. In cases where only one character state 

had expected values ≥5, the notation NA is used to indicate 

that the sample sizes are not appropriate (large enough) for 

statistical analysis. For the male–female data comparisons, 

the differences among state distributions are ((31 − 23) +

(8 − 0) + (69 − 69)) = 16%, below the threshold for observer error and thus are not analyzed further. Such results 

are due to two causes, which cannot be accurately separated: (1) the variation reflects actual sample variation and 

is not meaningful, or (2) the variation recorded does not 

reflect the sample variation but is due to observer error.

Decisions on whether it is appropriate to combine 

categories of age or sex are made on evaluation of all of 

1

2 3

4

5

6

FIGURE 1. Map showing locations of specimens used for intrasample analyses: 1, Costa Rica/Panama Border; 2, Panama Canal; 3, Lower Río 

Chucunaque, Panama; 4, Ilha do Maracá, Brazil; 5, Boca do Acre, Brazil; and 6, Cusco Amazónico, Peru.
8 • smithsonian contributions to zoology

the individual sample analyses results. In other words, an 

overall pattern among locality sample results is used to 

determine whether categories are appropriately combined 

or not for the intersample geographic variation analyses. 

This approach assumes that there is no geographic variation in whether characters vary by either age or sex (there 

are no data at the outset to support or refute this alternative). However, at a minimum, it is reasonable to assume 

that the three Middle American samples would likely 

demonstrate similar patterns of age or sex category variation as every herpetologist who has studied the Middle 

American members of the L. bolivianus complex in the 

last 50 years has thought they represent a single species. 

The following guidelines appear appropriate on the basis 

of examination of the overall results. If at least five of the 

samples have variation exceeding observer error and at 

least one of those samples is statistically significant, the 

categories being tested should be analyzed separately. If at 

least five of the samples do not exhibit variation exceeding observer error and the one that does is not statistically significant, the categories can be combined. All other 

instances require judgment based on whether chi-square 

results would likely be significant if sample sizes were increased (to realistic sample sizes).

Dorsal Pattern

The results of analyses by age and sex are shown in 

Table 5. The age results are equivalent in terms of the 

TABLE 2. Dorsal pattern state occurrences (by age and sex) within the Lower Río Chucunaque, Panama sample State A, uniform 

between interocular bar (if present) and sacrum; state B, single 

chevron between interocular bar and sacrum without additional 

spots lateral to dorsal chevron; state C, pattern complex with two 

dorsal chevrons and spotting all over dorsum, chevrons often 

elongate and fused with each other and interorbital bar; J, juvenile. A dash (–) indicates no specimens demonstrated the pattern.

State J J♂ J♀ ♂ ♀

A 2 – – 3 4

B 4 1 – – –

C 26 4 2 5 7

TABLE 3. Dorsal pattern state occurrences by age and sex, 

Lower Río Chucunaque, Panama sample. Numbers in parentheses are percentages by category (e.g., within juveniles). State A, 

uniform between interocular bar (if present) and sacrum; state B, 

single chevron between interocular bar and sacrum without additional spots lateral to dorsal chevron; state C, pattern complex 

with two dorsal chevrons and spotting all over dorsum, chevrons 

often elongate and fused with each other and interorbital bar.

Age Sex

State Juvenile Adult ♂ ♀

A 2 (5) 7 (37) 3 (23) 4 (31)

B 5 (13) 0 (0) 1 (8) 0 (0) 

C 32 (82) 12 (63) 9 (69) 9 (69)

TABLE 4. Chi-square occurrence values for character states of 

the dorsal pattern, Lower Río Chucunaque, Panama sample. 

State A, uniform between interocular bar (if present) and sacrum; state B, single chevron between interocular bar and sacrum without additional spots lateral to dorsal chevron; state C, 

pattern complex with two dorsal chevrons and spotting all over 

dorsum, chevrons often elongate and fused with each other and 

interorbital bar.

Number of juveniles Number of juveniles

State observed expected

A 2 6

B 5 3

C 32 30

TABLE 5. Age and sex intrasample analyses results for dorsal 

pattern. Definitions: >OE, observer error threshold passed; NS, 

chi-square results not statistically significant at ≥0.05 level; NA, 

data inadequate for chi-square analysis; c/w = compared with. 

A dash (–) indicates that it would be inappropriate to perform a 

significance test as the observer error threshold was not passed.

Juveniles c/w Adults Females c/w Males

Sample >OE Significance >OE Significance

Lower Río Chucunaque, Yes NS No –

Panama

Panama Canal Yes NS No –

Costa Rica/Panama Border No – Yes NS

Cuzco Amazónico, Peru Yes NS Yes NA

Boca do Acre, Brazil Yes NA No –

Ilha do Maracá, Brazil No – Yes NA
number 635 • 9

guidelines described above. The chi-square values for the 

three samples that exceed the observer error threshold are 

0.158, 0.094, and 0.094. It is reasonable to expect that 

these values could reach a 5% significance level if sample 

sizes were increased. Therefore dorsal patterns are analyzed separately by age (juveniles versus adults) in subsequent intersample analyses. Male and female results are 

also equivalent. The one sample that is appropriate for 

statistical testing has a chi-square value of 0.371. The two 

samples that exceed the observer error threshold that are 

not appropriate for statistical analysis could easily have 

comparable character state distributions with changing 

only 2–4 state conditions. In this case, it is not probable 

that increasing sample sizes would lead to statistically significant results. Male and female dorsal pattern data are 

combined for the intrasample variation summary (Table 6) 

and in subsequent intersample analyses.

Subocular Spot

Application of the guidelines to the age and sex analysis results (Table 7) indicates that juvenile and adult subocular spot states should be analyzed separately, whereas the 

male–female dichotomy is equivocal. The three chi-square 

values comparing the male and female data are 0.612, 

0.772, and 0.360. As it is improbable that realistically increasing sample size would result in statistical significance 

at the 5% level, the male and female data are combined 

for the intrasample variation summary (Table 6) and for 

subsequent intersample analyses.

Lip Stripe

The variation in results of the age and sex analyses 

(Table 8) mirrors that described for the subocular spot. 

Juvenile and adult data are best analyzed separately. The 

male and female category data are equivocal. The single 

chi-square test result for the Boca do Acre, Brazil data 

is 0.368. The distribution of occurrences of states from 

Cuzco Amazónico, Peru is almost identical and is rather 

similar for the Panama Canal sample. The male and female data are not differentiated for the intrasample summary (Table 6) or in subsequent intersample analyses.

Belly Pattern

The juvenile and adult analysis results (Table 9) are 

equivocal as to whether the data are best analyzed combined or separately. The three chi-square results involved 

are 0.348, 0.404, and 0.805. Realistically, increasing the 

sample sizes involved is unlikely to result in statistical significance. The juvenile and adult data are combined. Application of the guidelines for the male and female data 

analysis results (Table 9) yield combining of these categories as well for the intrasample summary (Table 6) and 

subsequent intersample analyses.

Thigh Pattern

Application of the guidelines to the age and sex analysis results (Table 10) indicates that both age and sex categories are best combined for the intrasample summary 

(Table 11) and in subsequent intersample analyses.

Shank Pattern

Application of the guidelines to the age and sex results indicated that both age and sex categories are best 

combined for the intrasample summary (Table 11) and in 

subsequent intersample analyses. None of the six samples 

analyzed met the observer error threshold for either age or 

sex analysis.

Dorsolateral Folds and Lateral Folds

Application of the guidelines to the age and sex analyses results for both characters (Tables 12, 13) indicate that 

age and sex categories are best combined for the intrasample summary (Table 11) and in subsequent intersample 

analyses.

Male Secondary Sexual Characters

The distributions of states among the samples are presented in Table 14.

Measurement Data

There is considerable variation in measurement value 

ranges within samples (Table 15). The intrasample variation in all variables far exceeds the measurement error 

variation.

There are unexpectedly large overlaps in body sizes 

(SVL) between specimens classified as juvenile and adult 

males and females. For the four samples with adequate 

data for males, the overlap in SVL between the smallest 

adult male and largest juvenile male ranges from 4.0 to 

18.0 mm. For the two samples with adequate data for 

females, the overlaps range from 8.8 to 22.0 mm. The 

largest values in overlap between juveniles and adults in 
1 0 • smithsonian contributions to zoology

TABLE 6. Occurrence (percent) of dorsal pattern, suborbital spot, lip pattern, and belly pattern states within samples by age and region. 

Dorsal pattern states: A, uniform between interocular bar (if present) and sacrum; B, single chevron between interocular bar and sacrum 

without additional spots lateral to dorsal chevron; C, pattern complex with two dorsal chevrons and spotting all over dorsum, chevrons 

often elongate and fused with each other and interorbital bar. Suborbital spot states: A, distinct; B, indistinct; C, absent. Lip pattern 

states: A, noticeable broad light stripe extending from front of snout to commissural gland, bordered below by continuous or broken 

markings on upper lip and either bordered above by dark line under eye or not; B, narrow light stripe from at least nostril to lower posterior end of eye; C, narrow light stripe from below front of eye through commissural gland; D, lip region uniform (about same intensity 

as background dorsal color) with or without a dark stripe under the eye and/or dark bars/mottling on the upper lip. Belly pattern states:

A, belly speckled, spotted, or mottled anteriorly only or with an anterior-posterior gradient; B, belly boldly mottled anteriorly only or 

with an anterior-posterior gradient; C, belly speckled or mottled with same intensity over entire belly; D, belly boldly mottled with same 

intensity over entire belly; E, belly without a pattern.

Dorsal pattern Suborbital spot Lip pattern Belly pattern

 Juvenile/Adult 

State Juveniles Adults State Juveniles Adults State Juveniles Adults State data combined

Costa Rica/Panama Border

(n = 45) (n = 27) (n = 45) (n = 27) (n = 45) (n = 27) (n = 72)

A 2 15 A 84 30 A 93 26 A 56

B 18 3 B 16 59 B 0 0 B 4

C 80 81 C 0 11 C 0 0 C 19

 D 7 74 D 19

 E 1

Panama Canal

(n = 29) (n = 17) (n = 29) (n = 17) (n = 29) (n = 17) (n = 45)

A 10 41 A 69 47 A 52 24 A 58

B 10 29 B 24 41 B 0 0 B 4

C 80 29 C 7 12 C 3 0 C 20

 D 45 76 D 7

 E 11

Lower Río Chucunaque, Panama

(n = 39) (n = 19) (n = 39) (n = 19) (n = 39) (n = 19) (n = 58)

A 5 37 A 64 90 A 51 37 A 50

B 13 0 B 31 10 B 0 0 B 0

C 82 63 C 5 0 C 8 0 C 45

 C/D 5 0 D 2

 D 36 63 E 3

Ilha do Maracá, Brazil

(n = 28 (n = 8) (n = 28) (n = 8) (n = 28) (n = 8) (n = 36)

A 11 25 A 71 50 A 7 0 A 25

B 25 12 B 0 0 B 0 0 A/B 3

C 64 62 C 29 50 C 0 0 B 39

 C/D 7 12 C 14

 D 86 88 D 19

 E 0

(continued)
number 635 • 1 1

this study noticeably exceed those from a study of the 

Leptodactylus podicipinus–L. wagneri complex (Heyer, 

1994:4–9). The overlaps are comparable for unpublished 

data for large species of the Leptodactylus pentadactylus cluster (e.g., L. knudseni juvenile-adult male SVL 

overlap 26.6 mm, juvenile-adult female SVL overlap 

11.0 mm; L. labyrinthicus juvenile-adult male SVL overlap 22.4 mm; juvenile-adult female SVL overlap 11.0 

mm; L. savagei juvenile-adult male SVL overlap 29.0 

mm.). These data suggest at least two nonexclusive explanations: (1) the morphological proxies used to differentiate adults from juveniles may not be consistently true and 

TABLE 6. (Continued)

Dorsal pattern Suborbital spot Lip pattern Belly pattern

 Juvenile/Adult 

State Juveniles Adults State Juveniles Adults State Juveniles Adults State data combined

Boca do Acre, Brazil

(n = 12) (n = 24) (n = 12) (n = 24) (n = 12) (n = 24) (n = 36)

A 8 4 A 42 83 A 17 42 A 83

B 33 71 B 0 0 B 0 0 B 0

C 58 25 C 58 17 C 58 17 C 0

 C/D 8 33 D 3

 D 17 8 E 14

Cuzco Amazónico, Peru

(n = 16) (n = 16) (n = 16) (n = 16) (n = 16) (n = 16) (n = 31)

A 6 31 A 88 62 A 93 50 A 90

B 19 44 B 12 31 B 0 0 B 0

C 75 25 C 0 6 C 7 25 C 10

 D 0 25 D 0

 E 0

TABLE 7. Age and sex intrasample analyses results for subocular spot. Definitions: >OE, observer error threshold passed; NS, 

chi-square results not statistically significant at ≥0.05 level; NA, 

data inadequate for chi-square analysis; c/w = compared with. 

A dash (–) indicates that it would be inappropriate to perform a 

significance test as the observer error threshold was not passed.

Juveniles c/w Adults Females c/w Males

Sample >OE Significance >OE Significance

Lower Río Chucunaque, Yes NS Yes NS

Panama

Panama Canal Yes NS No –

Costa Rica/Panama Border Yes 0.012 Yes NS

Cuzco Amazónico, Peru Yes NA Yes NA

Boca do Acre, Brazil Yes NA Yes NA

Ilha do Maracá, Brazil No – Yes NS

TABLE 8. Age and sex intrasample analyses results for lip stripe 

pattern. Definitions: >OE, observer error threshold passed; NS, 

chi-square results not statistically significant at ≥0.05 level; NA, 

data inadequate for chi-square analysis; c/w = compared with; 

a dash (–) indicates that it would be inappropriate to perform a 

significance test as the observer error threshold was not passed.

Juveniles c/w Adults Females c/w Males

Sample >OE Significance >OE Significance

Lower Río Chucunaque, Yes NS No –

Panama

Panama Canal Yes NS Yes NA

Costa Rica/Panama Border Yes 0.0004 No –

Cuzco Amazónico, Peru Yes NA Yes NA

Boca do Acre, Brazil Yes NS Yes NS

Ilha do Maracá, Brazil No – No –
1 2 • smithsonian contributions to zoology

(2) the variation observed in Leptodactylus is related to 

overall adult size.

INTERSAMPLE VARIATION

After mapping localities, 20 regional samples were chosen to analyze the nature of variation among them (Figure 

2). The sampling strategy was to maximize both geographic 

coverage and sample sizes. This strategy was not possible in 

some areas of coverage, and smaller than desired samples 

are included because of their importance in geographic representation or potential nomenclatural impact.

Patterns and Folds

The approach used to analyze intersample variation 

for the morphological characters involving patterns and 

folds follows that used in the analysis of intrasample variation. Any comparisons between samples are first analyzed 

to determine whether the amount of variation exceeds 

observer error. For the comparisons that exceed observer 

error, chi-square tests were used when appropriate to determine statistical significance. Juvenile and adult data are 

analyzed separately for dorsal pattern, subocular spot, and 

lip stripe characters. Juvenile and adult data are combined 

for analysis of belly pattern, thigh pattern, shank pattern, 

dorsolateral fold, and lateral fold characters.

Rather than make comparisons between all pairs of 

samples for the characters involved, three networks of 

samples were used because the purpose of this evaluation 

is to determine whether the characters demonstrate any 

patterns of geographic variation. There are three major 

geographic areas occupied by the Leptodactylus bolivianus complex: (1) Middle America, for which three mainland samples and two island samples, one Atlantic and 

one Pacific were selected; (2) populations predominantly 

occurring in South American river basins draining to the 

Caribbean for which eight samples were selected (the 

Northern Roraima sample is geographically near the rest 

of the samples but is in an Amazonian drainage area); and 

(3) populations occurring in river basins draining to the 

Amazon River, for which six samples were selected. Geographically proximate samples are compared among these 

three major geographic areas to determine overall patterns 

of differentiation.

The full set of analytic results are shown for the first 

character analyzed as an example of the comparisons that 

exceed observer error, those for which χ2

 analyses are inappropriate (only one state with an expected sample size of 5 

or greater), and the level and variation of χ2 values when χ2

values were appropriate. Results for the other characters 

are available from WRH on request and will be placed in 

the Smithsonian Institution Archives by 1 January 2012.

Dorsal Pattern

There are no significant differences among the three 

mainland Middle American samples (Table 16). The Gulf 

of Panama sample does not differ from the Panama Canal

TABLE 9. Age and sex intrasample analyses results for belly 

pattern. Definitions: >OE, observer error threshold passed; NS, 

chi-square results not statistically significant at ≥0.05 level; NA, 

data inadequate for chi-square analysis. A dash (–) indicates that 

it would be inappropriate to perform a significance test as the 

observer error threshold was not passed; c/w = compared with.

Juveniles c/w Adults Females c/w Males

Sample >OE Significance >OE Significance

Lower Río Chucunaque, No – No –

Panama

Panama Canal Yes NS Yes NS

Costa Rica/Panama Border Yes – No –

Cuzco Amazónico, Peru No NS No –

Boca do Acre, Brazil No NS No –

Ilha do Maracá, Brazil Yes – No –

TABLE 10. Age and sex intrasample analyses results for thigh 

pattern. Definitions: >OE, observer error threshold passed; NS, 

chi-square results not statistically significant at ≥0.05 level; NA, 

data inadequate for chi-square analysis; c/w = compared with. 

A dash (–) indicates that it would be inappropriate to perform a 

significance test as the observer error threshold was not passed.

Juveniles c/w Adults Females c/w Males

Sample >OE Significance >OE Significance

Lower Río Chucunaque, No – No –

Panama

Panama Canal No – No –

Costa Rica/Panama Border No – Yes NA

Cuzco Amazónico, Peru No – No –

Boca do Acre, Brazil No – No –

Ilha do Maracá, Brazil Yes NA No –
number 635 • 1 3

TABLE 11. Occurrence (percent) of thigh pattern, shank pattern, dorsolateral fold, and lateral fold states within region samples. Thigh 

pattern states: A, posterior thigh with a uniform or variable dispersion of melanophores (weakly to moderately mottled) usually for 

entire extent of posterior thigh; B, posterior thigh boldly mottled with large light irregular spots/vermiculations on a dark background 

to the light marks being more extensive than the dark areas, the light markings sometimes coalesced. Shank pattern states: A, uniform; 

B, short dark transverse bars extending less than halfway across dorsal surface of shank; C, at least one dark transverse bar extending 

halfway or more across the dorsal surface of the shank. Dorsolateral folds states: A, dorsolateral fold distinct and continuous from 

supratympanic fold to end of body, or with a brief interruption of the fold, or the fold distinct only from the supratympanic fold to the 

sacrum; B, dorsolateral fold with several interruptions to indescernable (including lack of fold due to poor preservation). Lateral folds 

states: A, lateral fold distinct or slightly interrupted from supratympanic fold to leg; B, lateral fold distinct only in groin region, indistinct overall, or indiscernible (including due to poor preservation).

Thigh pattern Shank pattern Dorsolateral folds Lateral folds

State Occurrence (%) State Occurrence (%) State Occurrence (%) State Occurrence (%)

Costa Rica/Panama Border

(n = 72) (n = 72) (n = 72) (n = 72)

A 17 A 7 A 71 A 29

A/B* 1 B 28 B 29 B 71

B 81 C 65 

Panama Canal

(n = 46) (n = 46) (n = 45) (n = 45)

A 0 A 2 A 67 A 29

A/B* 6 B 50 B 33 B 71

B 93 B/C* 2 

 C 46 

Lower Río Chucunaque, Panama

(n = 58) (n = 58) (n = 58) (n = 58)

A 0 A 0 A 81 A 57

A/B* 3 B 24 B 19 B 43

B 97 C 76 

Ilha do Maracá, Brazil

(n = 36) (n = 36) (n = 36) (n = 36)

A 8 A 0 A 100 A 0

A/B* 14 B 42 B 0 B 100

B 78 B/C* 3 

 C 56 

Boca do Acre, Brazil

(n = 36) (n = 36) (n = 36) (n = 33)

A 100 A 3 A 97 A 30

B 0 B 78 B 3 B 70

 C 19 

Cuzco Amazónico, Peru

(n = 32) (n = 32) (n = 32) (n = 32)

A 100 A 0 A 100 A 12

B 0 B 38 B 0 B 88

 C 62 

*Indicates a condition that is intermediate between the two states.
1 4 • smithsonian contributions to zoology

sample but does differ from the other two mainland samples. The (Pacific) Gulf of Panama sample does not differ 

from the (Caribbean) San Andrés/Providencia sample. The 

San Andrés/Providencia sample additionally does not differ from the Panama Canal sample, but does differ from 

the Southern Costa Rica/Eastern Panama and Darién

samples.

The geographically adjacent Darién and Northern 

Magdalena Drainage samples do not differ from each 

other (Table 16).

Among the primarily Caribbean drainage samples, 

there are only two statistically significant results: juvenile 

Northern Guyana data compared with juvenile Suriname

data (Table 16 sample comparisons K:M) and juvenile 

Northern Guyana data compared with juvenile Northern 

Roraima data (Table 16 sample comparisons K:N). As 

there are 72 comparisons within this network of samples, 

having two statistically significant results is well within 

statistical expectations.

There are no statistically significant differences between the Meta Drainage and the Amazonian drainage 

samples (Rio Solimões/Amazonas, Río Ucayali, Madre de 

Dios, Lower Rio Purús/Rio Acre, Middle/Lower Río Madeira, South Central Bolivia) (Table 16). There is only one 

statistically significant difference in the juvenile data for 

the Northern Roraima–South Central Bolivia comparison (Table 16, sample comparisons N:T). There are no 

compelling data to favor the Meta Drainage sample or the 

Northern Roraima sample as showing a pattern in common with the rest of the Amazonian drainage samples.

There are no statistically significant differences among 

the Amazonian drainage network of samples (Table 16).

Subocular Spot

There are no demonstrable differences among juveniles in the Middle American network samples. The adult 

data show differences between the mainland samples 

Southern Costa Rica/Eastern Panama compared with the 

Darién data (χ2

 = 0.001), and the Panama Canal sample 

compared with the Darién data (χ2 = 0.020). The adult 

data also differ between the San Andrés/Providencia sample and the Southern Costa Rica/Eastern Panama sample 

(χ2 = 0.035) and with the Darién sample (χ2 = 0.001).

The adult data differ between the samples that geographically link the Darién sample with the Northern 

Magdalena Drainage sample (χ2

 = 0.044). Juvenile data 

are distinct for the following comparisons: MiddleSouthern Magdalena Drainage sample compared with the 

Northern Roraima Sample (χ2 = 0.033); Meta Drainage

sample compared with the Northern Roraima sample (χ2

= 0.006); Southern Guyana sample compared with the 

Suriname sample (χ2 = 0.022); and the Southern Guyana

sample compared with the Northern Roraima sample (χ2 = 

0.004). Adult data are distinct for the following comparisons: Northern Magdalena Drainage sample compared 

with the Meta Drainage sample (χ2 = 0.042), Meta Drainage sample with the Southern Guyana sample (χ2

 = 0.003), 

Meta Drainage sample with the Suriname sample (χ2 = 

0.009), and the Meta Drainage sample with the Northern 

TABLE 12. Age and sex intrasample analyses results for dorsolateral folds. Definitions: >OE, observer error threshold passed; NS, 

chi-square results not statistically significant at ≥0.05 level; NA, 

data inadequate for chi-square analysis; c/w = compared with. 

A dash (–) indicates that it would be inappropriate to perform a 

significance test as the observer error threshold was not passed.

Juveniles c/w Adults Females c/w Males

Sample >OE Significance >OE Significance

Lower Río Chucunaque, Yes NS No –

Panama

Panama Canal No – Yes NS

Costa Rica/Panama Border Yes NS No –

Cuzco Amazónico, Peru No – No –

Boca do Acre, Brazil No – No –

Ilha do Maracá, Brazil No – No –

TABLE 13. Age and sex intrasample analyses results for lateral 

folds. Definitions: >OE, observer error threshold passed; NS, 

chi-square results not statistically significant at ≥0.05 level; NA, 

data inadequate for chi-square analysis; c/w = compared with. 

A dash (–) indicates that it would be inappropriate to perform a 

significance test as the observer error threshold was not passed.

Juveniles c/w Adults Females c/w Males

Sample >OE Significance >OE Significance

Lower Río Chucunaque, Yes NS No –

Panama

Panama Canal No – Yes NS

Costa Rica/Panama Border No – No –

Cuzco Amazónico, Peru No – Yes NA

Boca do Acre, Brazil No – No –

Ilha do Maracá, Brazil No – No –
number 635 • 1 5

Roraima sample (χ2 = 0.027). Overall, the Meta Drainage

sample stands out as being distinctive within this network 

of samples.

The geographically network-linking Meta Drainage

sample differs from both the Rio Solimões/Amazonas

sample (adult data, χ2 = 0.006) and the Madre de Dios

sample (adult data, χ2 = 0.004). The Northern Roraima

sample differs from the Madre de Dios sample (juvenile 

data χ2 = 0.011), the Lower Purús/Rio Amazonas sample 

(adult data, χ2 = 0.003), and the South Central Bolivia

TABLE 14. Male secondary sexual character state occurrences (percent) within samples. Arm states: A, arm not hypertrophied; B, arm 

hypertrophied. Lower jaw tubercle states: A, absent; B, tuberculate. Chest states: A, no patch of tubercles; B, central patch of tubercles; 

C, lateral patches of tubercles in addition to central patch. Thumb spine states: A, one modest chisel-shaped spine; B, one hypertrophied 

chisel-shaped spine; C, two round spines. Tympanum states: A, annulus lacking tubercles; B, annulus tuberculate. A dash (–) indicates 

no state or no percent occurrence for samples with n = 0.

Arm hypertrophy Jaw tubercles Chest tubercles Thumb spines Tympanum tubercles

State Value State Value State Value State Value State Value

Costa Rica–Panama Border

n = 15 n = 12 n = 15 n = 15 n = 15

A 33 A 67 A 53 A 0 A 0

B 67 B 33 B 47 B 0 B 100

 C 0 C 100 

Panama Canal

n = 11 n = 10 n = 11 n = 11 n = 11

A 64 A 80 A 64 A 0 A 0

B 36 B 20 B 36 B 0 B 100

 C 0 C 100 

Lower Río Chucunaque, Panama

n = 8 n = 7 n = 8 n = 8 n = 8

A 50 A 57 A 38 A 0 A 25

B 50 B 43 B 62 B 0 B 75

 C 0 C 100 

Ilha do Maracá, Brazil

n = 1 n = 0 n = 1 n = 1 n = 0

A 100 – – A 100 A 100 – –

B 0 – – B 0 B 0 – –

 C 0 C 0 

Boca do Acre, Brazil

n = 4 n = 2 n = 4 n = 4 n = 4

A 75 A 50 A 100 A 25 A 0

B 25 B 50 B 0 B 75 B 100

 C 0 C 0 

Cuzco Amazónico, Peru

n = 5 n = 4 n = 5 n = 5 n = 4

A 0 A 25 A 20 A 0 A 0

B 100 B 75 B 20 B 100 B 100

 C 60 C 0
1 6 • smithsonian contributions to zoology

TABLE 15. Intrasample variation in measurement data. Definitions: n, sample size; A, Lower Río Chucunaque, Panama sample; B, 

Panama Canal sample; C, Costa Rica–Panama Border sample; D, Cuzco Amazónico, Peru sample; E, Boca do Acre, Brazil sample; F, 

Ilha do Maracá, Brazil sample; Min, minimum value (mm); Max, maximum value (mm); Range, range of values (mm); SVL, snout–vent 

length; HL, head length; HW, head width; EN, eye–nostril distance; TD, tympanum diameter. A dash (–) indicates there is only one 

individual in the sample.

Males Females

Variable Sample n Min Max Range n Min Max Range

SVL A 9 78.9 97.9 19.0 16 75.5 89.3 13.8

B 11 67.8 101.3 33.5 6 79.0 95.5 16.5

C 15 76.0 92.8 16.8 12 74.5 86.8 12.3

D 5 106.2 116.0 9.8 11 77.9 99.8 21.9

E 4 79.0 97.4 18.4 20 63.0 86.3 23.3

F 1 – 86.0 – 7 66.0 95.5 29.5

HL A 9 29.1 36.2 7.1 16 28.4 32.0 3.6

B 11 25.2 34.7 9.5 6 28.1 35.1 7.0

C 15 28.4 33.3 4.9 11 26.4 32.4 6.0

D 5 36.6 40.4 3.8 11 29.3 34.4 5.1

E 4 28.7 37.3 8.6 20 24.0 32.7 8.7

F 1 – 32.5 – 7 25.5 34.0 8.5

HW A 9 27.1 36.9 9.8 16 25.0 29.2 4.2

B 11 23.3 36.5 13.2 6 26.3 32.7 6.4

C 15 26.0 33.1 7.1 12 25.0 28.6 3.6

D 5 35.2 40.4 5.2 11 23.8 31.5 7.7

E 4 24.2 31.0 6.8 20 20.0 28.1 8.1

F 1 – 29.5 – 7 22.0 31.0 9.0

EN A 9 8.5 9.5 1.0 16 7.7 9.4 1.7

B 11 7.4 9.9 2.5 6 7.8 9.8 2.0

C 15 8.0 9.8 1.8 12 7.5 9.0 1.5

D 5 11.0 11.7 0.7 11 8.3 9.7 1.4

E 4 7.8 9.8 2.0 20 6.5 8.7 2.2

F 1 – 9.0 – 7 7.0 9.0 2.0

TD A 9 5.5 7.1 1.6 16 5.2 6.2 1.0

B 11 5.0 7.8 2.8 6 5.6 6.6 1.0

C 15 5.2 6.4 1.2 12 5.2 6.3 1.1

D 5 6.8 8.5 1.7 11 6.0 7.2 1.2

E 4 5.8 7.6 1.8 20 4.5 6.5 2.0

F 1 – 6.0 – 7 5.0 6.5 1.5

Thigh A 9 37.3 43.0 5.7 15 32.3 41.0 8.7

B 11 30.4 48.7 18.3 6 36.3 46.7 10.4

C 15 33.8 42.0 8.2 12 30.9 39.2 8.3

D 5 42.8 56.7 13.9 11 36.1 45.0 8.9

E 4 37.8 47.1 9.3 20 30.0 39.8 9.8

F 1 – 46.5 – 7 30.5 46.0 15.5

Shank A 9 40.8 46.7 5.9 15 37.2 44.3 7.1

B 11 34.8 49.0 14.2 6 41.3 48.9 7.6

C 15 38.1 45.6 7.5 12 36.6 43.5 6.9

D 5 51.6 58.5 6.9 11 40.6 48.2 7.6

E 4 41.5 51.3 9.8 20 33.5 47.5 14.0

F 1 – 46.5 – 7 36.5 50.0 13.5

Foot A 9 39.8 46.6 6.8 16 38.1 44.4 6.3

B 11 37.2 48.8 11.6 6 40.2 50.2 10.0

C 15 37.8 45.5 7.7 12 37.6 44.7 7.1

D 5 53.5 60.3 6.8 11 42.8 50.2 7.4

E 4 43.5 52.4 8.9 20 35.0 46.6 11.6

F 1 – 48.0 – 7 37.0 49.0 12.0
number 635 • 1 7

sample (juvenile data, χ2 = 0.001). Both the Meta Drainage

and Northern Roraima samples are weakly differentiated 

(at best) from the remaining Amazonian drainage samples.

Within the Amazonian drainage network exclusive of 

the Northern Roraima sample, the following comparisons 

are distinct: Rio Solimões/Rio Amazonas with the Lower 

Rio Purús/Rio Acre samples (adult data, χ2

 = 0.012); 

Madre de Dios with the South Central Bolivia samples 

(juvenile data, χ2 = 0.003); and the Lower Rio Purús/Rio 

Acre with the South Central Bolivia samples (adult data, 

χ2 = 0.032).

Lip Stripe

There are only two statistically significant different 

comparisons within the Middle American network of 

samples: Southern Costa Rica/Eastern Panama with the 

Panama Canal samples (juvenile data, χ2

 = 0.016) and the 

Southern Costa Rica/Eastern Panama with the Darién

samples (juvenile data, χ2 = 0.010).

The data do not statistically differ between the 

network-linking Darién and Northern Magdalena Drainage samples.

A

B C

D

E

F

G

H

I

J

K

L

M

N

O

P

Q

R

S

T

FIGURE 2. Map showing locations of 20 regional samples. A, Costa Rica/Panama Border; B, Panama Canal; C, Darién; D, Gulf of Panama; 

E, San Andrés/Providencia; F, Northern Magdalena Drainage; G, Middle/Southern Magdalena Drainage; H, Santa Marta; I, Meta Drainage; 

J, Caracas; K, Northern Guyana; L, Southern Guyana; M, Suriname; N, Northern Roraima; O, Rio Solimões/Amazonas; P, Río Ucuyali; Q, 

Madre de Dios; R, Lower Rio Purus/Rio Acre; S, Middle/Lower Rio Madeira; and T, South Central Bolivia.
1 8 • smithsonian contributions to zoology

TABLE 16. Intersample comparisons for dorsal pattern data. Significant chi-square (χ2) values are in italic font and followed by an asterisk. Sample definitions: A, Southern Costa Rica/Eastern Panama; B, Panama Canal; C, Darién; D, Gulf of Panama; E, San Andrés/

Providencia; F, Northern Magdalena Drainage; G, Middle/Southern Magdalena Drainage; H, Santa Marta; I, Meta Drainage; J, Caracas; K, Northern Guyana; L, Southern Guyana; M, Suriname; N, Northern Roraima; O, Rio Solimões/Amazonas; P, Río Ucayali; Q, 

Madre de Dios; R, Lower Rio Purús/Rio Acre; S, Middle/Lower Rio Madeira; T, South Central Bolivia. Other abbreviations: n, sample 

size; NA, samples not appropriate for chi-square analysis. A dash (–) indicates that it was inappropriate to perform a significance test as 

the observer error threshold was not passed.

Juvenile data Adult data Juvenile data Adult data

Sample Exceeds Exceeds Sample Exceeds Exceeds

pair observer observer pair observer observer

comparisons n error χ2 n error χ2 comparisons n error χ2 n error χ2

A–B 49, 31 No – 29, 21 Yes 0.111

A–C 49, 79 No – 29, 49 Yes 0.102

B–C 31, 79 No – 21, 49 Yes 0.343

A–D 49, 2 Yes 0.691 49, 21 Yes 0.021*

B–D 31, 2 Yes 0.324 21, 21 Yes 0.296

C–D 79, 2 Yes 0.793 49, 21 Yes 0.029*

D–E 2, 7 Yes NA 21, 29 Yes 0.292

A–E 49, 7 Yes NA 29, 29 Yes 0.007*

B–E 31, 7 Yes NA 21, 29 No –

C–E 79, 7 Yes NA 49, 29 Yes 0.009*

C–F 79, 20 Yes 0.390 49, 23 No –

F–G 20, 75 No – 23, 85 No –

F–H 20, 64 No – 23, 45 Yes 0.313

F–I 20, 6 Yes NA 23, 12 Yes 0.638

F–J 20, 30 No – 23, 27 No –

F–K 20, 22 Yes 0.338 23, 14 Yes 0.497

F–L 20, 39 No – 23, 18 Yes 0.497

F–M 20, 15 Yes 0.222 23, 13 Yes 0.781

F–N 20, 46 No – 23, 23 No –

G–H 75, 64 No – 23, 45 Yes 0.411

G–I 75, 6 Yes 0.978 23, 12 Yes 0.792

G–J 75, 30 No – 23, 27 No –

G–K 75, 22 Yes 0.186 23, 14 Yes 0.663

G–L 75, 39 No – 23, 18 Yes 0.507

G–M 75, 15 No – 23, 13 Yes 0.838

G–N 75, 46 No – 23, 23 Yes 0.866

H–I 20, 6 Yes 0.963 23, 12 Yes 0.524

H–J 20, 30 No – 23, 27 Yes 0.556

H–K 20, 22 Yes 0.174 23, 14 Yes 0.505

H–L 20, 39 No – 23, 18 Yes 0.152

H–M 20, 15 No – 23, 13 Yes 0.451

H–N 20, 46 No – 23, 23 Yes 0.630

I–J 6, 30 Yes 0.915 12, 27 Yes 0.343

I–K 6, 22 Yes 0.758 12, 14 No –

I–L 6, 39 No – 12, 18 Yes 0.480

I–M 6, 15 Yes NA 12, 13 Yes 0.352

I–N 6, 46 No – 12, 23 No –

J–K 30, 22 Yes 0.486 27, 14 Yes 0.235

J–L 30, 39 No – 27, 18 Yes 0.214

J–M 30, 15 Yes 0.541 27, 13 Yes 0.628

J–N 30, 46 No – 27, 23 Yes 0.614

K–L 22, 39 Yes 0.155 14, 18 Yes 0.434

K–M 22, 15 Yes 0.042* 14, 13 Yes 0.248

K–N 22, 46 Yes 0.036* 14, 23 Yes 0.700

L–M 39, 15 Yes 0.619 18, 13 No –

L–N 39, 46 No – 18, 23 Yes 0.646

M–N 15, 46 Yes 0.481 13, 23 Yes 0.754

I–O 6, 9 Yes NA 12, 9 Yes NA

I–P 6, 18 No – 12, 8 Yes NA

I–Q 6, 73 No – 12, 19 Yes 0.857

I–R 6, 13 No – 12, 29 Yes 0.260

I–S 6, 2 Yes NA 12, 8 Yes NA

I–T 6, 51 Yes NA 12, 7 Yes NA

N–O 46, 9 Yes 0.582 23, 9 Yes 0.662

N–P 46, 18 No – 23, 8 Yes 0.597

N–Q 46, 73 Yes 0.092 23, 19 No –

N–R 46, 13 No – 23, 29 Yes 0.098

N–S 46, 2 Yes 1.000 23, 8 Yes 0.814

N–T 46, 51 Yes 0.033* 23, 7 Yes 0.875

O–P 9, 18 Yes NA 9, 8 Yes NA

O–Q 9, 73 Yes 0.356 9, 19 Yes 0.706

O–R 9, 13 Yes 0.545 9, 29 Yes 0.600

O–S 9, 2 No – 9, 8 Yes NA

O–T 9, 51 Yes 0.304 9, 7 Yes NA

P–Q 18, 73 No – 8, 19 Yes 0.414

P–R 18, 13 No – 8, 29 Yes 0.226

P–S 18, 2 Yes 1.000 8, 8 Yes NA

P–T 18, 51 Yes 0.221 8, 7 Yes NA

Q–R 73, 13 Yes 0.483 19, 29 Yes 0.475

Q–S 73, 2 Yes 0.743 19, 8 No –

Q–T 73, 51 No – 19, 7 No –

R–S 13, 2 Yes NA 29, 8 No –

R–T 13, 51 Yes 0.304 29, 7 Yes 0.814

S–T 2, 51 Yes 0.637 8, 7 No –
number 635 • 1 9

The data statistically differ between several of the primarily Caribbean draining network samples: Northern 

Magdalena Drainage with Southern Guyana (adult data, 

χ2 = 0.018); Northern Magdalena Drainage with Suriname

(adult data, χ2 = 0.018); Middle/Southern Magdalena 

Drainage with Northern Guyana (adult data, χ2 = 0. 018); 

Santa Marta with Northern Guyana (juvenile data, χ2 < 

0.001, adult data, χ2 = 0.001); Santa Marta with Southern 

Guyana (juvenile data, χ2 = 0.003, adult data, χ2 = 0.025); 

Santa Marta with Suriname (adult data, χ2 = 0.025); Meta 

Drainage with Northern Guyana (adult data, χ2 = 0.002); 

Meta Drainage with Southern Guyana (adult data, χ2 = 

0.035); Caracas with Northern Guyana (juvenile data, 

χ2 = 0.009; adult data, χ2 = 0.012); Northern Guyana with 

Southern Guyana (adult data, χ2 = 0.035); Northern Guyana with Northern Roraima (juvenile data, χ2

 < 0.001; 

adult data, χ2 = 0.001); Southern Guyana with Northern 

Roraima (juvenile data, χ2 < 0.001), and Suriname with 

Northern Roraima (adult data, χ2 = 0.053). The amount 

of demonstrable variation in this network clearly exceeds 

any variation that could be accounted for by chance alone.

The network-linking Meta Drainage sample differs 

from the Madre de Dios sample (adult data, χ2 = 0.052) 

and the Lower Rio Purús/Rio Acre sample (juvenile data, 

χ2 = 0.018; adult data, χ2 = 0.021). The Northern Roraima

sample differs from the Rio Solimões/Rio Amazonas sample (adult data, χ2

 = 0.031), Madre de Dios (juvenile data, 

χ2 < 0.001; adult data, χ2 = 0.035), Lower Rio Purús/Rio 

Acre (juvenile data, χ2 = 0.054; adult data, χ2 < 0.001), and 

South Central Bolivia (juvenile data, χ2 < 0.001) samples. 

There seems to be more differentiation between the Northern Roraima sample with the other Amazonian drainage 

samples than between the Meta Drainage sample and the 

primarily Amazonian network samples.

Within the primarily Amazonian drainage sample network, there are only two statistically significant comparisons: the Río Ucayali Drainage sample with the Lower 

Rio Purús/Rio Acre sample (juvenile data, χ2 = 0.010), and 

the Lower Rio Purús/Rio Acre sample with the South Central Bolivia sample (juvenile data, χ2

 = 0.040).

Belly Pattern

There are no significant differences among the Middle 

American samples or between the network-linking Darién

with the Northern Magdalena Drainage samples.

There are several statistically significant different comparisons within the primarily Caribbean drainage network 

samples: Northern Magdalena Drainage with Northern 

Guyana (χ2 = 0.002), Northern Magdalena Drainage

with Southern Guyana (χ2 = 0.001), Northern Magdalena 

Drainage with Northern Roraima (χ2 < 0.001), Middle/

Southern Magdalena Drainage with Northern Guyana

(χ2 = 0.004), Middle/Southern Magdalena Drainage with 

Southern Guyana (χ2 < 0.001), Middle/Southern Magdalena Drainage with Northern Roraima (χ2

 < 0.001), Santa 

Marta with Northern Guyana (χ2 = 0.010), Santa Marta

with Southern Guyana (χ2 = 0.001), Santa Marta with 

Northern Roraima (χ2 < 0.001), and Caracas with Southern Guyana (χ2

 = 0.030). In general, there is a contrast 

between coastal and river valley samples from Colombia 

and Venezuela with the Guyana and Northern Roraima 

samples.

There are no statistically significant differences between the network-linking Meta Drainage sample with the 

Amazonian network samples. In contrast, there are three 

statistically significant differences among the Northern 

Roraima and other Amazonian network samples: Madre 

de Dios (χ2

 < 0.001), Lower Rio Purús/Rio Acre (χ2 = 

0.001), and South Central Bolivia (χ2 < 0.001).

There are no statistically significant differences among 

the primarily Amazonian network samples.

Thigh Pattern

There are no significant differences among the Middle 

American samples or between the network-linking Darién

with the Northern Magdalena Drainage samples.

There are statistically significant differences between 

several of the primarily Caribbean network samples: 

Northern Magdalena Drainage with Meta Drainage (χ2 = 

0.003), Northern Magdalena Drainage with Southern 

Guyana (χ2 < 0.001), Middle/Southern Magdalena Drainage with Southern Guyana (χ2

 < 0.001), Santa Marta with 

Meta Drainage (χ2

 = 0.034), Santa Marta with Southern 

Guyana (χ2 < 0.001), Meta Drainage with Caracas (χ2 = 

0.005), Meta Drainage with Northern Guyana (χ2 = 

0.016), Meta Drainage with Suriname (χ2 = 0.015), Caracas with Southern Guyana (χ2

 < 0.001), Northern Guyana with Southern Guyana (χ2

 < 0.001), Southern Guyana

with Suriname (χ2 < 0.001), Southern Guyana with Northern Roraima (χ2

 < 0.001).

Each comparison between the network linking Meta 

Drainage and Northern Roraima samples is statistically 

significant with the primarily Amazonian network samples: Meta Drainage with Rio Solimões/Rio Amazonas

(χ2 = 0.027), Río Ucayali (χ2 = 0.011), Madre de Dios (χ2 = 

0.001), Lower Rio Purús/Rio Acre (χ2 = 0.001), Middle/
2 0 • smithsonian contributions to zoology

Lower Rio Madeira (χ2 = 0.004), and South Central Bolivia (χ2

 = 0.001) samples; Northern Roraima with Rio 

Solimões/Rio Amazonas (χ2 = 0.008), Rio Ucayali (χ2 < 

0.001), Madre de Dios (χ2 < 0.001), Lower Rio Purús/Rio 

Acre (χ2 < 0.001), Middle/Lower Rio Madeira (χ2 = 0.048), 

and South Central Bolivia (χ2 < 0.001) samples.

Within the primarily Amazonian network samples 

only the Río Ucayali and Madre de Dios samples differ 

significantly (χ2 = 0.022). 

The thigh pattern variation demonstrates the most 

geographically coherent pattern (Figure 3).

Shank Pattern

There are no statistically significant differences among 

the Middle American samples or between the networklinking Darién with the Northern Magdalena Drainage

samples.

There are four statistically significant differences 

among the primarily Caribbean drainage network samples: 

Northern Magdalena Drainage with Northern Guyana

(χ2

 = 0.009), Meta Drainage with Northern Guyana (χ2 = 

0.017), Caracas with Northern Guyana (χ2 = 0.004), and 

FIGURE 3. Variation in posterior thigh patterns among 20 regional samples. Pattern A, light gray; pattern B, white; intermediate pattern A/B, 

dark gray.
number 635 • 2 1

Northern Guyana with Northern Roraima (χ2 = 0.003). The 

Northern Guyana sample stands out within this network.

There is one statistically significant difference between the network-linking Meta Drainage sample with 

the Lower Rio Purús/Rio Acre sample (χ2 = 0.011). There 

are no statistically significant differences between the 

network-linking Roraima sample with the other Amazonian samples.

There are two statistically significant differences 

among the primarily Amazonian drainage network samples: Madre de Dios with Lower Rio Purús/Rio Acre (χ2

 = 

0.005) and Lower Rio Purús/Rio Acre with South Central 

Bolivia (χ2 = 0.016).

Dorsolateral Folds

There are no statistically significant differences among 

the Middle American samples or between the networklinking Darién with the Northern Magdalena Drainage

samples.

There are several statistically significant differences 

among the primarily Caribbean drainage network samples: Northern Magdalena Drainage with Santa Marta

(χ2 = 0.010), Northern Magdalena Drainage with Caracas

(χ2 < 0.001), Northern Magdalena Drainage with Southern Guyana (χ2

 = 0.002), Northern Magdalena Drainage

with Suriname (χ2 = 0.023), Northern Magdalena Drainage with Northern Roraima (χ2

 < 0.001), Middle/Southern 

Magdalena Drainage with Northern Roraima (χ2 = 0.013), 

and Northern Guyana with Northern Roraima (χ2 = 

0.033). The distinctiveness of the Northern Magdalena 

Drainage sample is due to more occurrences of the dorsolateral folds with several interruptions to indiscernible 

conditions: it is possible that this is due to poor preservation rather than a real sample difference.

There are no statistically significant differences between the network-linking Meta Drainage and Northern 

Roraima samples with the rest of the Amazonian drainage 

network samples.

There are no statistically significant differences between any of the Amazonian network samples.

Lateral Folds

There are no statistically significant differences among 

the Middle American samples or between the networklinking Darién with the Northern Magdalena Drainage

samples.

The following primarily Caribbean draining network 

samples differ statistically significantly: Middle/Southern 

Magdalena Drainage with Northern Roraima (χ2

 = 0.013), 

Santa Marta with Northern Guyana (χ2 = 0.023), Santa 

Marta with Northern Roraima (χ2 < 0.001), Caracas with 

Northern Guyana (χ2 = 0.009), and Caracas with Northern Roraima (χ2

 = 0.003).

The network-linking Meta Drainage sample statistically differs from the Río Ucayali sample (χ2

 = 0.025). 

The network-linking Northern Roraima sample statistically differs from the Río Ucayali (χ2

 = 0.007), Lower Rio 

Purús/Rio Acre (χ2 = 0.048), and South Central Bolivia

(χ2 = 0.013) samples.

There are three sample comparisons that statistically 

differ significantly within the Amazonian network: Río 

Ucayali with Madre de Dios (χ2 = 0.005), Madre de Dios

with Lower Rio Purús/Rio Acre (χ2 = 0.030), and Madre 

de Dios with South Central Bolivia (χ2

 = 0.005).

Male Secondary Sexual Characters

Sample sizes are inadequate to perform statistical 

tests for many of the sample comparisons. In addition, the 

problem of seasonal development of sexual characters has 

to be addressed. Comparisons among samples should be 

made on males that are sexually active and have developed their full extent of secondary sexual characteristics. 

For this purpose, the presence of vocal slits is inappropriate, as once the slits are developed, they are not lost and 

regained on a seasonal basis. For comparative purposes, 

any noticeable arm hypertrophy is used as a proxy for full 

development of secondary sexual characters, realizing that 

this proxy is approximate and not definitive.

The data are best analyzed visually with the raw numbers of state occurrences for all 20 geographic samples 

(Table 17).

There are two obvious features in the data. First, there 

is no coherent pattern to the state occurrences for the jaw 

tubercle and tympanic annulus tubercle characters. The 

most reasonable interpretations for these two characters 

are that (1) tympanic annulus tubercles, once developed, 

are not shed seasonally; and (2) jaw tubercles are shed seasonally and are redeveloped each year by all adult males 

in all populations. Second, there is geographically coherent variation among the states of the chest tubercles and 

thumb spine characters. As can be seen from the raw data 

(Table 17), several of the samples have individuals with 

only a central patch of chest tubercles (state A) as well as 

individuals with central and lateral patches of chest tubercles (state B). This variation is interpreted to be ontogenetic; that is, for individuals that have both central and 

lateral tubercle patches, the central patch develops before 
2 2 • smithsonian contributions to zoology

the lateral patches develop on an annual basis. This interpretation is supported by comparison of the jaw tubercle 

and chest tubercle data, assuming that both jaw and chest 

tubercles start developing simultaneously each season in 

a given male. Thus the critical aspect for this character 

is whether any individuals in a population exhibit both 

central and lateral tubercle patches. It is assumed that 

if any individuals in a population have both central and 

lateral patches, all individuals in that population will develop both. The data appear to demonstrate a geographic 

break between the Caracas and Northern Guyana samples 

(Table 17). Samples for which all males lack chest tubercles provide no information on whether males develop 

lateral patches (Rio Purús/Rio Acre and Lower/Middle 

Madeira samples). The only adult male in the Río Ucayali

sample did not have noticeably hypertrophied arms. The 

geographically most incongruous sample is the Suriname

sample, where all three males with chest tubercles have 

only central patches.

Variation in thumb spines indicates three groupings 

of the geographic samples. The first is the contrast of two 

versus one spine, with the Costa Rica/Eastern Panama

through Caracas samples having two spines (see Figure 

13A) and all other samples characterized by having one 

spine. There is also variation among the one spine samples whether the spine is slightly chisel shaped (see Figure 13C) or obviously chisel shaped (see Figure 13B). The 

Río Ucayali sample male lacking arm hypertrophy has a 

medium-sized slightly chisel-shaped spine, suggesting that 

the spine is not fully developed.

Measurement Data

Discriminant function analyses were performed on the 

20 geographic samples separately for the female and male 

data. The data sets were run with SYSTAT 11 software 

(Engelman et al., 2004) using the complete, forward, and 

backward stepwise models. The results for the complete, 

forward, and backward stepwise models are very similar; 

only the complete model results are discussed.

The first canonical vector accounts for 63% of the 

cumulative dispersion for the female data, the first two 

canonical vectors account for 81%, and 90% is reached 

with the third canonical vector. None of the 20 samples 

are completely correctly identified in the posterior classification matrix using all data nor in the jackknife classification matrix. The poor discrimination of the data set 

is reflected in the plot of the first two canonical vectors 

(Figure 4). Three distinctive sample groups can be distinguished from each other and the rest of the samples: 

(1) the San Andrés/Providencia and Northern Magdalena 

Drainage samples, (2) the Gulf of Panama sample, and (3) 

the Southern Guyana sample (Figure 4).

The first canonical vector accounts for 55% of the cumulative dispersion for the male data, the first two canonical vectors account for 75%, and 90% is reached between 

the fourth and fifth canonical vectors. All individuals in 

the Río Ucayali and South Central Bolivia samples were 

posteriorly correctly identified in the full data classification matrix, but all individuals from any sample were not 

correctly posteriorly classified in the jackknife classification matrix. The plot of the first two canonical vectors 

shows the same level of poor discrimination of the samples 

TABLE 17. Number of males exhibiting secondary sexual character states. Number of occurrences may not be equal among 

characters due to missing observations. Samples are defined 

in Table 16. Jaw tubercle states: A, absent; B, present. Chest 

tubercle states: A, absent; B, central patch; C, central and lateral patches. Thumb spine states: A, 1 slightly chisel-shaped; B, 

1 robustly chisel-shaped; C, 2 round. Tympanic tubercle states:

A, absent; B, present. A dash (–) indicates no specimens had the 

character state involved.

Jaw Chest Thumb Tympanic 

tubercles tubercles spines tubercles

Sample A B A B C A A/B* B C A B

A 3 5 2 8 0 0 0 0 10 0 10

B 1 5 1 6 0 0 0 0 7 0 7

C 0 5 0 10 0 0 0 0 10 0 10

D 2 3 1 6 0 0 0 0 7 0 7

E 2 10 4 13 0 0 0 0 16 0 14

F 3 1 3 6 0 0 0 0 9 1 8

G 6 26 2 36 0 0 0 0 42 1 41

H 1 3 4 9 0 0 0 0 13 0 13

I – – 1 3 0 0 0 0 3 0 3

J 2 3 2 6 0 0 0 0 8 0 8

K 4 3 4 1 2 4 3 0 0 0 7

L 1 0 1 0 1 2 0 0 0 0 2

M 1 3 3 3 0 4 2 0 0 1 5

N – – 0 1 3 4 0 0 0 0 4

O 0 1 0 0 1 0 0 1 0 0 1

P – – – – – – – – – – –

Q 1 3 3 0 2 0 0 6 0 0 5

R – – 1 0 0 0 0 1 0 0 1

S – – 2 0 0 0 0 2 0 0 2

T 0 2 1 0 1 0 0 2 0 0 2

*In the case of thumb spines, there were some individuals that exhibited 

intermediate states between state A and state B.
number 635 • 2 3

as the female data (results not shown). There are three 

samples that are distinct from each other and the rest of 

the samples: (1) Gulf of Panama, (2) Northern Guyana, 

and (3) Madre de Dios.

The measurement data indicate that the Gulf of Panama sample is the most distinctive as its distinctiveness 

appears in both the female and male data analyses.

GEOGRAPHIC VARIATION AMONG THE 

SAMPLES

Adult Form Morphological Characters

For the characters analyzed statistically (including discriminant function analyses results), within the three networks, there is a moderate trend of greater differentiation 

between geographically distant sample pairs, as would 

be expected. In terms of evaluating geographic variation, 

the most insightful patterns of differentiation are those 

between adjacent geographic samples. The patterns are 

best visualized by comparing moderate to high (differing 

by three to five characters) levels of differentiation between geographically adjacent samples (Figure 5). Overall, 

there is considerable differentiation among adjacent sample pairs that are geographically distant from each other.

Taking all the differentiation data together (Figure 5), 

the most significant break between geographically adjacent samples is between the Caracas and Northern Guyana samples. These two samples differ from each other by 

four statistically meaningful characters, and the males of 

the Caracas sample lack lateral patches of chest tubercles 

and have two rounded thumb spines whereas the Northern 

Guyana sample males have lateral patches of chest spines 

and have one slightly chisel-shaped thumb spine. This Caracas sample break is discrete from the Northern Guyana

sample and is most consistent with species level differentiation. The Caracas sample character distribution pattern 

continues along coastal Venezuela to the Estado de Sucre 

and the Island of Trinidad. There were few locality samples 

analyzed between southern Anzoategui State in Venezuela 

and the Guyana and Venezuelan border along the Estado de 

Bolívar (Venezuela), most of which lacked sexually active 

adult males. The closest Venezuelan locality sample with 

an adult male with noticeably hypertrophied arms with 

two round thumb spines is from Bolívar State, Santo Tomé 

de Guayana, San Felix (AMNH 81456) and an adult male 

with a single slightly chisel-shaped thumb spine is from 

-5 -4 -3 -2 -1 0 1 2 3 4 5

Factor 1

-5

-4

-3

-2

-1

0

1

2

3

4

Factor 

2

1-A

1-B

2

3

FIGURE 5. Moderate (3 characters) to high (4 or 5 characters) levels 

of differentiation between geographically adjacent samples.

FIGURE 4. Discriminant function analysis results of first and second 

canonical vectors for female measurement data of 20 geographic 

samples (denoted by various symbols). The Northern Magdalena 

Drainage (1-A, triangle with apex at left)/San Andrés/Providencia

(1-B, triangle with apex at bottom), Gulf of Panama (2, triangle with 

apex at top), and Southern Guyana (3, narrow vertical bar) samples 

are distinctive from each other.
2 4 • smithsonian contributions to zoology

Bolívar State, 13 km south, 1 km east Puente Cuyuni (KU 

166408), over 250 km distant from each other. Additional 

data are needed from the Venezuelan states of Amazonas 

and Bolívar to determine the precise distribution of the two 

species involved and whether they occur in sympatry.

For the northeast set of samples (Southern Costa Rica/

Eastern Panama through Caracas) the level of variation 

is consistent with intraspecific variation, although there 

is considerable variation between the Darien and Gulf of 

Panama samples (Figure 5C, D).

In contrast, there is much more variation among the 

remaining samples (Figure 5). A clear geographic pattern is demonstrated by whether the male thumb spine is 

strongly chisel shaped and the posterior thigh predominantly uniform or moderately mottled (Rio Solimões/

Amazonas through the Río Ucayali and South Central 

Bolivia samples) or a weakly chisel-shaped thumb spine 

and a predominantly boldly mottled posterior thigh pattern (Northern Roraima through Suriname samples). The 

morphological data are equivocal in terms of whether the 

break between this clustering of samples represents intraor interspecific variation.

Larval Morphology

Given the problems of tadpole identification described 

in Methods and Materials, justification concerning the 

identification of the larval samples described below is 

called for. Five species of Leptodactylus occur in Costa 

Rica: L. insularum, L. fragilis, L. melanonotus, L. savagei, 

and L. poecilochilus. All five Costa Rican species have 

published larval descriptions and figures (L. insularum as

L. bolivianus, L. savagei as L. pentadactylus, L. fragilis 

as L. labialis; Heyer 1970), and the tadpoles of the five 

species can readily be identified based on morphological 

features. Only L. melanonotus larvae might be confused 

with L. insularum larvae. The dorsal oral papilla gap of 

L. insularum is narrower (21–31% of oral disk) than the 

gap in L. melanonotus from Costa Rica (40–60%). The 

L. insularum larval samples from Isla San Andrés (Colombia) do not have any noticeable differences with respect to 

the L. insularum from Costa Rica.

Four larval samples are available in addition to the 

description by Heyer (1970) for this species. All materials share the following features: sinistral spiracle, median 

vent, entire oral disk, a single or double row of oral disk 

marginal papillae, a tooth row formula of 2/3, and dorsal 

fin origin at the tail-body juncture. The larval materials are 

from the following localities, referred to henceforth by the 

specific localities only: USNM 241340: Isla San Andrés, 

San Andrés y Providencia, Colombia; USNM 330406: 

near Cañas, Guanacaste, Costa Rica; KU 68365: Puerto 

Cortes, Puntarenas, Costa Rica; and USNM 330837: 

Rincón de Osa, Puntarenas, Costa Rica.

The two specimens from Isla San Andrés and the 

sample from Puerto Cortés are faded. The dorsal bodies 

of the Cañas tadpoles are relatively uniform brown; the 

guts are very visible from the lateral view with the portion anterior to the guts relatively uniform tan/brown; the 

ventral aspect of the body is lighter than the dorsum, but 

melanophores are present on the portion anterior to the 

guts; the tail musculature is uniform brown; the tail fins 

are much lighter than the musculature but either with scattered melanophores or a relatively uniform tan pattern. 

The Puerto Cortés tadpole dorsal aspects of the head and 

bodies are mottled brown; from the side, the head-bodies 

are mottled; in ventral view, the area of the oral disk to the 

guts has scattered melanophores, with the area over the 

guts without melanophores; the tail fins and musculature 

are dark, almost uniform, brown. 

There is variation among larval quantitative character 

data (Table 18).

TABLE 18. Quantitative larval data. Number in parentheses under Source is number of specimens upon which the data are based. Definitions: ED, eye diameter; HBL, head–body length; ODW, oral disk width; TL, total length; TN, number of A2 teeth in 0.3 mm. A dash 

(–) indicates the values were not available in the data source.

Locality Source Gosner stages TL, mm HBL/TL, % ED/HBL, % ODW/BL, % TN

Colombia, Isla San Andrés USNM 241340 (2) 37 32.6–35.2 32–34 11–12 27–28 17–19

Costa Rica Heyer, 1970 34 32.0 32–44 5–10 20–28 –

Costa Rica, Cañas USNM 330406 (5) 28–31 24.4–28.5 38–42 6–7 23–27 18–22

Costa Rica, Puerto Cortés KU 68365 (5) 32–34 29.3–32.4 37–39 6–7 21–23 18–20

Costa Rica, Rincón de Osa USNM 330837 (5) 25–30 17.4–18.4 44–47 8–10 20–23 25–28
number 635 • 2 5

The variation of the data described above is difficult 

to evaluate in terms of discerning intraspecific from interspecific differentiation. At least some of the quantitative 

variation (Table 18) should be correlated with ontogeny 

as represented by Gosner developmental stages. Head 

and body lengths were measured differently for the Heyer 

(1970) data than for the data presented here, which compromises comparability. If one makes the reasonable assumption that all of the Costa Rican data represent a 

single species, those data pretty much encompass all of the 

variation observed among all the larval data.

Advertisement Calls

There are only three relatively noisy recordings on 

hand for members of the L. bolivianus-insularum complex suitable for analysis in addition to the information 

published by Fouquette (1960) (Table 19).

Costa Rica Recording

Figure 6

Recording KU 801, as indicated previously, has a noticeably erratic tape speed, which adds an error factor to 

the quantitative data. The call consists of a single note 

composed of 1 or 2 pulses. The recording has two call 

rates, in the beginning 19 s the call rate is 1.2 calls/s, in the 

last 39 s, the call rate is 2.5 calls/s. The first half of the call 

has a sharper rising frequency modulation than the last 

half of the call. The highest frequency occurs at the end of 

the call or shortly before the end of the call (at about 85% 

total call duration). There is considerable variation in 

dominant frequencies among the calls analyzed doubtless 

due to tape speed variation; the most consistent values are 

those reported in Table 19. Harmonic structure is present, 

but not strongly developed.

Panama Recording

Figure 7

The Barro Colorado Nature Monument recording has 

calls consisting of single notes. There is sufficient other 

sound in the broadcast channel that the amplitude intensity 

of the call is not definitively clear; most calls have evidence 

of two primary pulses per note. The first portion of the note 

(approximately 30%) has a faster rise of frequency than the 

next 50% of the note. The final approximate 20% of the 

note has a noticeable falling frequency. Harmonic structure 

is present and stronger than in the KU 801 recording.

Peru Recording

Figure 8

Recording USNM 268 cut 11 has calls consisting of 1 

or 2 notes. The quality of the recording is not sufficient to 

evaluate detailed temporal structure. The analyses suggest 

that the second note for two note calls or the single note 

of one note calls have at least two pulses. The first 20% 

(approximately) of the call has a sharply rising frequency, 

with a decreased rate of rising frequency for the remainder 

of the call. The highest frequency is consistently at the end 

of the call. There is no visible harmonic structure in the 

audiospectrogram analyses of the calls; there is too much 

noise in the wave form analyses to determine with certainty whether there is harmonic structure or not.

TABLE 19. Advertisement call data. Definitions: ID, identification; SVL, snout–vent length. A dash (–) indicates values not available 

from the data source.

 Voucher No. Call Beginning Highest Ending Dominant

Recording Temperature, SVL, notes/ duration, frequency, frequency, frequency, frequency,

Country ID °C mm call Calls/s s Hz Hz Hz Hz Harmonics

Costa Rica KU 801 26 – 1 1.2–2.5 0.08 220–300 1,080–1,200 1,060–1,200 920–1,130 +

Panama BCI CD – – 1 1.5 0.10–0.12 110–180 890–1,100 510–600 740–870 +

Panama Fouquette, 24 – – 1.9 0.10 200 900 750 – +

1960

Peru USNM 268, 23.5 112 1–2 0.5 0.12–0.16 130–200 730–780 730–780 600–690 ?

cut 11
2 6 • smithsonian contributions to zoology

The available advertisement call data (Table 19) are 

most consistent with two call types representing distinct 

species: (1) the recordings from Costa Rica and Panama; 

and (2) the recording from Peru.

MOLECULAR RELATIONSHIPS

A total of 800 aligned base pairs were included in 

the analyses, of them 540 were constant and 260 variable 

(172 parsimony informative). Base composition consisted 

of A = 34%, C = 23%, G = 18%, and T = 25%. Thirty-one 

(3.87%) positions included an indel position in at least 

one taxon.

All analyses recovered a clade that included all representative samples of the L. latrans species group (Figure 9). Within this clade, a subclade consisting of L. latrans

and L. chaquensis is recovered and is the sister group to 

the L. insularum-bolivianus complex clade. Within the 

L. insularum-bolivianus complex, three subclades are 

recovered: subclade A, including samples from Panama, 

Venezuela, and Colombia, corresponds to the taxon 

known as L. insularum; subclade B, found in Peru and 

Bolivia, represents the taxon known as L. bolivianus; and 

subclade C is recognized herein as a new species in this 

complex occurring in the Guiana Shield region.

The genetic divergences among the four subclades 

recovered within the L. latrans species complex range 

from 9.5 to 14.1% (see Figure 9 for range of divergence 

values followed by the mean value below corresponding 

branches). Mean value divergences among the three species 

recognized in the L. bolivianus–insularum complex from 

other species of the L. latrans species group range from 4.2 

to 7.2%. Divergences among the three species recognized 

in the L. bolivianus–insularum complex are bolivianus–

insularum 4.9%; insularum–sp. nov. 6.2%; and bolivianus–sp. nov. 5.1%. In addition, mean divergences among 

the sampled species of the latrans species group are insularum–latrans = 12.2%; insularum–chaquensis = 12.1%; 

bolivianus–latrans = 10.1%; bolivianus–chaquensis = 

S

-20

-10

10

20

30

0.001

19.466 19.5 19.52 19.54 19.56 19.58 19.6 19.62

1

2

3

4

5

0.000

19.466 19.6 19.7 19.8 19.9

Pressur

e Kilohertz

Time in seconds

FIGURE 6. Advertisement call of Leptodactylus, KU recording 801 from just west of Piedras Blancas, 

Puntarenas, Costa Rica, 8 April 1966, water temperature 78°F.
number 635 • 2 7

10.8%; latrans–sp. nov. =12.9%; chaquensis–sp. nov. = 

12. 6%; and latrans–chaquensis = 6.8%. The existence of 

an undescribed taxon demonstrated by the genetic diversification recovered in this analysis in combination with 

the subsequent identification of diagnostic morphological 

characters supports the recognition of the new species. 

The genetic diversification among this complex of species 

is consistent with the level of diversification among the 

other two species in the L. bolivianus complex as well as 

level of diversification between species in the L. latrans

and those in the L. bolivianus complex.

INTERSPECIFIC VARIATION

Three outlying localities for which there are only juvenile specimens available were considered problematic 

as to species identification in the preceding analyses. As 

previously indicated, Marinus Hoogmoed (pers. comm.) 

informed WRH at the VI World Congress of Herpetology in 2008 that he had reexamined frogs from the state 

of Pará, near the state of Amapá and found them to be 

members of the Leptodactylus bolivianus complex. These 

specimens belong to the new species recognized herein. 

Thus the single specimen from Amapá examined in this 

study is considered to represent the new species as well.

There are also a number of localities between the distributions of L. bolivianus and the new species for which 

only juveniles and/or females are available and rather 

arbitrary criteria were used to assign species identifications. The following localities are considered to represent 

L. bolivianus. COLOMBIA. Vaupes: Río Querari near 

junction of Río Querari and Río Vaupés. BRAZIL. Amazonas: Parque Nacional Pico da Neblina, São Gabriel da 

Cachoeira; Rio Cuieiras, entering Rio Negro on left bank; 

Rio Demeni, Barcelos; São Gabriel da Cachoeira; São 

FIGURE 7. Advertisement call of Leptodactylus from Barro Colorado Nature Monument CD recording, track 46, Panama.

S

-20

20

0.001

6.74 6.8 6.85 6.9

1

2

3

4

5

0.000

6.74 7 7.2 7.4 7.6 7.8

Pressur

e Kilohertz

Time in seconds
2 8 • smithsonian contributions to zoology

João, near Tapurucuara. VENEZUELA. Amazonas: Catarata de Huá (Salto de Huá); Río Casiquiare, 8 miles [12.9 

km] below Orinoco (Canal Casiquiare); Río Mavaca, 108 

km SSE Esmeralda. Presumed localities for the new species 

follow. BRAZIL. Roraima: Uiacás, Rio Urariquera; Serra 

dos Surucucus; Missão Catrimani; Cachoeira do Cujubim, 

Rio Catrimani; Parque Nacional Viruá, próximo a Caracaraí; Praia do Guariúba; Santa Maria do Buiaçu. The 

comparisons that follow include the preceding locality/

specimen data, and even if most presumed identifications 

are incorrect, the overall results would not be meaningfully impacted.

Measurement Data

Complete model discriminant function analyses of the 

measurement data for females and males indicate incomplete separation of the three species, with the male data 

(Figure 10) showing better discrimination than the female 

data (Figure 11).

For the male analysis, the first canonical vector accounts for 84% of the cumulative dispersion and the first 

two canonical vectors account for 100% of the cumulative 

dispersion. Eighty-three percent of L. bolivianus, 92% of 

L. insularum, and 85% of the new species are correctly 

identified using all data in the posterior classification matrix. The male data jackknifed classification matrix for 

L. bolivianus, L. insularum, and Leptodactylus, new species are 78, 92, and 79% respectively.

For the female data discriminant analysis, the cumulative proportions of total dispersion are identical to the 

male data (84 and 100%). Seventy-one percent of L. bolivianus, 87% of L. insularum, and 80% of Leptodactylus, new species are correctly identified using all data in the 

posterior classification matrix. The female data jackknifed 

classification matrix is 67, 84, and 78% respectively.

The measurement data indicate that there is differentiation among the three species, but the differentiation is 

not complete enough to be useful as diagnostic information to correctly identify all specimens to species.

FIGURE 8. Advertisement call of Leptodactylus, USNM recording 268 cut 11 from Tambopata, 

Madre de Dios, Peru.

U S

-200

200

0.50

14.755 14.8 14.85 14.9

kHz S

1

2

3

4

5

0.000

14.755 15 15.2 15.4 15.6 15.8

Pressur

e Kilohertz

Time in seconds
number 635 • 2 9

Morphological Data

Forty-four percent of the morphological character 

comparisons among species pairs are compromised by 

high observer error (Table 20). The high observer error is 

accounted for, in part, by the fact that the three species are 

rather morphologically similar to each other.

For the statistically significant character comparisons, the most differentiated character states are listed as 

follows.

FIGURE 9. Maximum likelihood cladogram of the Leptodactylus latrans species group with other representatives of 

Leptodactylus species lineages.

-5 -2 1 4

FACTOR(1)

-5

-2

1

4

FACTOR(2)

FIGURE 10. Plot of first two canonical vectors for male measurement data 

for the three species of the Leptodactylus bolivianus cluster. Dot (●), L. bolivianus; ×, L. insularum; +, new species. Centroid confidence ellipses, p = 0.95.
3 0 • smithsonian contributions to zoology

Leptodactylus bolivianus and 

L. insularum Comparisons

Dorsal pattern A: uniform between interocular bar 

[if present] and sacrum; L. insularum with greater occurrence. Lip stripe pattern D: lip region uniform; L. bolivianus with greater occurrence. Posterior thigh patterns A 

and B: both species differing by 86% for each state. Belly 

pattern C: belly speckled or mottled with same intensity 

over entire belly; L. insularum with greater occurrence. 

Thumb spines state C: two round spines; L. insularum

with 100% occurrence.

Leptodactylus bolivianus: 

New Species Comparisons

Posterior thigh pattern A: uniform or variable dispersion of melanophores (weakly to moderately mottled) usually for entire extent of posterior thigh; L. bolivianus with 

greater occurrence. Belly pattern A: speckled, spotted, or 

mottled anteriorly only or with an anterior-posterior gradient; L. bolivianus with greater occurrence. Chin tubercles: both states equally differentiated in the two species. 

Chest tubercles state B: central patch only; new species 

with greater occurrence. Thumb spines, state B: one hypertrophied chisel-shaped spine; L. bolivianus with exclusive occurrence.

Leptodactylus insularum: 

New Species Comparisons

Belly pattern B: boldly mottled anteriorly only or with 

an anterior-posterior gradient; new species with greater 

occurrence. Lateral folds, both states equally differentiated in both species. Chest tubercles state B: central patch 

of tubercles only; new species with greater occurrence. 

Thumb spines, state A: one modest chisel-shaped spine 

on each thumb; new species with greater occurrence. The 

male secondary sexual characteristics demonstrate the 

greatest diagnostic differentiation among the three species.

SPECIES ACCOUNTS

There are several characters examined in this study 

that demonstrate negligible differences among the three 

species. All three species exhibit the following character 

states, which are not repeated in the individual species 

accounts.

Dorsal patterns are uniform between the interocular 

bar (if present) and the sacrum or with a single chevron 

between the interocular bar and sacrum without additional 

spots lateral to the dorsal chevrons or a complex pattern 

with two dorsal chevrons and spotting all over the dorsum, 

chevrons often elongate and fused with each other and the 

interorbital bar. Light subocular patches vary from very 

distinct to not discernible. The dorsal surface of the shank 

patterns are uniform or speckled or with short dark transverse bars extending less than halfway across the shank or 

with at least one dark transverse bar extending halfway 

or more across the shank or with large dark spots in the 

middle of the upper shank. Dorsolateral folds are distinct 

and continuous from the supratympanic fold to the end 

of the body, or with a brief interruption of the fold, or the 

fold is distinct only from the supratympanic fold to the 

sacrum, or the fold has several interruptions, or the fold is 

indiscernible. The toes are fringed. Adult males have either 

keratinous tubercles on the tympanum or lack tubercles.

Specimens from two localities cannot be identified 

to species: USNJM 162697, 216724–216725, Venezuela, Bolívar, Los Patos, 25–28 km SE El Manteco, 350 m, 

7°11N, 62°22W; and UMMZ 195839, Venezuela, Amazonas, San Fernando de Atabapo, 4°3N, 67°28W. The 

specimens involved are metamorphs, juveniles, or females. 

FIGURE 11. Plot of first two canonical vectors for female measurement data for the three species of the Leptodactylus bolivianus cluster. Circle, L. bolivianus; ×, L. insularum; +, new species. Centroid 

confidence ellipses, p = 0.95.

-4.0 -1.8 0.4 2.6 4.8 7.0

FACTOR(1)

-4.0

-1.8

0.4

2.6

4.8

7.0

FACTOR(2)
number 635 • 3 1

There are additional localities for which the morphological data are ambiguous as to which species is involved for 

samples geographically between the new species and L. insularum. The identifications for these localities are based 

on river drainage distributions, and new data are needed 

to verify which of the two species occur at the localities 

involved. These latter localities are distinguished on the 

distribution map (see Figure 14) and in the distribution 

sections of the following pertinent accounts.

Leptodactylus bolivianus Boulenger, 1898

Figure 12

Leptodactylus bolivianus Boulenger, 1898:131. Lectotype: Museo Civico 

di Storia Naturale de Genova 28875, male, designated by Capocaccia 

(1957:214). Lectotype locality: Barraca (Río Madidi, Bolivia).

Leptodactylus romani Melin, 1941:54. Lectotype Göteborgs Naturhistoriska 

Museum (GNM) 499, juvenile, designated by Heyer (1969:6). Lectotype locality: Taracuá, Rio Uaupés, Brazil.

TABLE 20. Statistical comparison of morphological character states for species pairs of the Leptodactylus bolivianus complex; df = 

degrees of freedom. Significant probability (p) results are in bold italic font.

Leptodactylus bolivianus Leptodactylus bolivianus Leptodactylus insularum

Characters and L. insularum and L. new species and L. new species

Dorsal pattern χ2 = 25.186 Character state Character state 

df = 4 occurrence < observer error occurrence < observer error

p = 0.000

Lip stripe χ2 = 33.945 Character state Character state

df = 3 occurrence < observer error occurrence < observer error

p = 0.000

Subocular spot χ2 = 2.443 χ2 = 1.306 χ2 = 3.711

df = 2 df = 2 df = 2

p > 0.1 p > 0.5 p > 0.1

Tibia pattern Character state Character state Character state

occurrence < observer error occurrence < observer error occurrence < observer error

Posterior thigh pattern χ2

 = 238.297 χ2 = 152.059 Character state

df = 2 df = 2 occurrence < observer error

p = 0.000 p = 0.000

Belly pattern χ2 = 25.186 χ2 = 76.760 χ2 = 55.100

df = 4 df = 4 df = 4

p = 0.000 p = 0.000 p = 0.000

Dorsolateral folds Character state Character state Character state

occurrence = observer error occurrence < observer error occurrence < observer error

Lateral folds Character state Character state χ2 = 10.281

occurrence = observer error occurrence < observer error df = 1

 p = <0.005

Chin tubercles χ2 = 0.035 χ2 = 8.607 χ2 = 0.489

df = 1 df = 1 df = 1

p > 0.5 p = 0.000 p > 0.5

Chest tubercles χ2 = 0.096 χ2 = 7.747 χ2 = 7.281

df = 1 df = 2 df = 2

p > 0.1 p < 0.025 p < 0.05

Thumb spines χ2 = 18.040 χ2 = 16.333 χ2 = 20.5

df = 1 df = 1 df = 1

p = 0.000 p = 0.000 p = 0.000

Tympanum tubercles Character state Character state Character state

occurrence < observer error occurrence < observer error occurrence < observer error
3 2 • smithsonian contributions to zoology

Etymology. There is no stated etymology in 

the original description. The species is named for the country of Bolivia from which the specimens were collected. 

Diagnosis. Most specimens of L. bolivianus

have the posterior thigh with a uniform or variable dispersion of melanophores (weakly to moderately mottled) 

usually for the entire extent of the posterior thigh; most 

specimens of L. insularum and L. n. sp. have boldly mottled 

posterior thighs with large light irregular spots/vermiculations on a dark background to the light marks being more 

extensive than the dark areas, the light markings sometimes 

coalesced. Leptodactylus bolivianus sexually active adult 

males have lateral patches of chest tubercles in addition to 

a central patch; L. insularum only have the central patch 

of tubercles. The only entirely consistent character diagnosing the three species is the adult male thumb spine. The 

single spine on each thumb in L. bolivianus is extremely 

chisel shaped (Figure 13B); L. insularum has two rounded, 

pointed spines per thumb (Figure 13A); L. n. sp. has a single, modestly chisel-shaped spine per thumb (Figure 13C).

FIGURE 12. Photo of Leptodactylus bolivianus from Tambopata, 

Peru (courtesy of Reginald B. Cocroft).

FIGURE 13. Male thumb spines. A. Leptodactylus insularum, USNM 565028; B. Leptodactylus 

bolivianus, USNM 280211; C. Leptodactylus new species, USNM 566172.

A B

C

A B

C
number 635 • 3 3

Character Summary. The most frequent 

lip stripe pattern consists of a noticeable broad light stripe 

extending from the front of the snout to the commissural 

gland, bordered below by continuous or broken markings 

on the upper lip and either bordered above by a dark line 

under the eye or not. The most frequent posterior thigh 

pattern is a uniform or variable dispersion of melanophores 

(weakly to moderately mottled) usually for the entire extent of the posterior thigh. The most frequent lateral fold 

conditions are distinct folds only in the groin region, or 

indistinct overall, or indiscernible. The most frequent adult 

male chin tubercle condition is presence of the tubercles.

Female (n = 91) SVL 61.2–107.7 mm (m [mean] = 

85.3), male (n = 25) 79.0–121.5 mm (m = 104.6). Female 

head length/SVL ratio 0.33–0.40 (m = 0.365), male 0.34–

0.41 (m = 0.365). Female head width/SVL ratio 0.29–0.37 

(m = 0.323), male 0.31–0.39 (m = 0.345). Female eyenostril distance/SVL ratio 0.09–0.12 (m = 0.102), male 

0.09–0.11 (m = 0.102). Female tympanum distance/SVL 

ratio 0.06–0.10 (m = 0.074), male 0.06–0.08 (m = 0.071). 

Female thigh length/SVL ratio 0.40–0.53 (m = 0.462), 

male 0.39–0.51 (m = 0.458). Female shank length/SVL 

ratio 0.44–0.56 (m = 0.516), male 0.46–0.56 (m = 0.502). 

Female foot length/SVL ratio 0.47–0.60 (m = 0.528), male 

0.48–0.57 (m = 0.526).

Tadpole. Unknown.

Advertisement Call. Calls of 1–2 notes at a 

rate of 0.5/s; call duration 0.12–0.16 s; call amplitude modulated, each note with loudest portion in middle of note; 

call frequency modulated starting at a low frequency (130–

200 Hz) followed by a long portion of higher frequency 

(730–780 Hz) (Figure 8). Recording USNM 268 cut 11.

Distribution. Central and western portions 

of the Amazon basin in Bolivia, Brazil, Colombia, Peru, 

and Venezuela (Figure 14).

Specimens Examined (n = 347). Localities enclosed by square brackets require validation that the 

species is correctly identified. BOLIVIA. Beni: Alejandria, 

Río Mamoré, 12°4S, 65° 9W, AMNH 79053–79066; 

Beni Biosphere Reserve, El Trapiche/Palm camp, 300 m, 

14°30S, 66°0W, USNM 306617; Ivon, 11°7S, 66°9W, 

UMMZ 58837; Puerto Almacen (= Mayor Pedro Vaca 

Diez), 260 m, 14°47S, 64°51W, AMNH 72381; Río 

Grande, 5 km NW of mouth, 15°51S, 64°39W (coordinates for mouth of Río Grande), AMNH 79042, 79052; 

Río Mamoré, Río Ibarre (= Ibare), 14°37S, 64°57W, 

AMNH 79033–79041, 79044–79055; Río Mapiri, 

mouth, below upper Beni, UMMZ 58834 (3), 58835 (2); 

Rurrenabaque, 220 m, 14°28S, 67°34W, MCZ-A 10091, 

UMMZ 58836 (3), 58838 (4); Santa Rosa, Río Mamoré, 

15°38S, 64°37W, AMNH 72382–72395; Tumi Chucua, 

170 m, 10°8S, 66°10W, USNM 280211. Cochabamba:

Puerto Chipiriri, 300 m, 16°32S, 65°16W (coordinates 

for mouth of Río Chipiriri), AMNH 72238–72241; Puerto 

San Francisco, 36 km N Chipiriri, 16°30S, 65°20W, 

KU 135508; Villa Tunari, 10.5 km E, 500 m, 16°59S, 

65°20W, KU 183010–183011. Pando: Nacebe, 11°0S, 

67°25W, NMP6V 72179/3, NMP6V 72179/4. Santa Cruz:

Buenavista (= Buena Vista), 400 m, 17°27S, 63°40W, 

UMMZ 66482, 66485, 66535, 83101 (3); Buena Vista, 

5 km W, USNM 146513–146514, 146522; Río Chaparé, 

5 km N boca, 15°55S, 64°38W, AMNH 79046–79047; 

Río Ichilo, 34 km S boca Río Chaparé, 16°07S, 64°49W, 

AMNH 79043; Río Ichilo, ~54 km S boca Río Chaparé, 

16°10S, 64°40W, AMNH 79048–79051.

BRAZIL. Acre: Feijó, 8°9S, 70°21W, MZUSP 

106665–106667; Igarapé de Nico, Rio Acre, 9°49S, 

67°36W, USNM 202431; Reserva Florestal Humaitá, 

9°45S, 67°40W, MZUSP 70866; Rio Branco, 60 m, 

9°58S, 67°48W, MZUSP 6487; Rio Muru, 8°09S, 

70°45W, MZUSP 106673–106675; Tarauacá, 190 m, 

8°10S, 70°46W, FMNH 83238, MZUSP 64730,106668–

106672. Amazonas: Lago do Miuá, Anavilhanas, 3°42S, 

62°8W, INPA 1232; [Barcelos, Rio Demeni, 1°0S, 

62°58W, INPA 9613]; Boca do Acre, 8°45S, 67°24W, 

MZUSP 50230–50248, USNM 202432–202448; Borba, 

25 m, 4°25S, 59°35W, MNRJ 3169, 13522, MZUSP 

56499–56501, USNM 202451–202453; Fortaleza, 

médio Purus, 6°26S, 64°17W, MZUSP 4474, 4480, 

8141; Humaitá, San Miguel, 70 m, 7°31S, 63°2W 

(coordinates for Humaitá), MNRJ 19295–19296; Igarapé Belém, Rio Solimões, 3°55S, 69°37W, AMNH 

97077–97079; [Rio Cuieiras, 2°50S, 60°30W, MZUSP 

65391, 65394 (metamorphs)]; [São Gabriel da Cachoeira, 

Parque Nacional Pico da Neblina, 0°5S, 65°55W, AL 

2015, INPA 15634, 15637]; [São João, próximo Tapurucuara, 0°33S, 64°57W, MZUSP 37509]; Xerua, lado 

direito, Rio Juruá, 6°8S, 67°50W, INPA 2248. Rondônia: Abunã, Rio Madeira, 9°42S, 65°23W, AL 122; 

Calama, 50 m, 8°4S, 62°52W, USNM 202455; Foz do 

Jamari, 8°28S, 63°30W, MZUSP 56504–56505; USNM 

202449–202450; Porto Velho, 98 m, 8°46S 63°55W, 

INPA 15458, MZUSP 16657; São Carlos, 100 m, 8°28S, 

63°30W, USNM 202454.

COLOMBIA. Amazonas: Leticia, ~100 m, 4°9S, 

69°57W, AMNH 81859–81862, KU 124742, USNM 

140289, 147029–147030, ICNMHN 46855; Leticia, 

Parque Nacional Natural Amacayacó, Cabaña Matamata, 

3°58S, 70°10W, IND-AN 2100; Puerto Nariño, 3°45S, 

70°22W, KU 153296–153297. Vaupés: [Río Querari near 
3 4 • smithsonian contributions to zoology

junction of Río Querari and Río Vaupés, 1°4N, 69°51W, 

USNM 193874 (21.4 mm juvenile)].

PERU. Locality uncertain, possible Iquitos, AMNH 

42890. Huánuco: Tournavista, 1200 m, 8°5556S, 

74°4219W, MCZ-A 75025. Loreto: near Contamana, 

Rean Rean, Lago Suhuayo, Río Ucuyali, 7°15S, 74°54W, 

AMNH 42899, 43416; Domo Santa Clara, Orellana, ~100 

m, 6°54S, 75°4W, USNM 127192–127194; Explorama 

Lodge, junction Río Yanomono and Río Amazonas, 

KU 220363; Iquitos, 100 m, 3°46S, 73°15W, AMNH 

42570, MCZ-A 100080; Pampas Hermosas (= Pampa 

Hermosa), Río Cushabatay, 7°12S, 75°18W, AMNH 

42141, 42147, 42831, 43120; Roaboya, ~100 m, 7°48S, 

74°52W, AMNH 42603–42604, 42920, 45532; “San 

Antonio,” Río Itaya, 3°56S, 73°43W, AMNH 43214. 

Madre de Dios: Cocha Cashu, Río Manú between Río 

Panagua and Río Cachira, ~400 m, 11°51S, 71°19W, KU 

154880–154885; Cuzco (= Cusco) Amazonico, 200 m, 

12°35S, 69°5W, KU 194911, 204508, 205143–205168, 

207732–207734, 209182; Itahuanía, 450 m, 12°47S, 

71°13W, FMNH 81533; Las Pampas del Río Heath, 

160 m, 13°0S, 69°0W, USNM 314902; Pakitza, 350 m, 

11°57S, 71°17W, USNM 342635–342640; Puerto 

Maldonado, 189 m, 12°36S, 69°11W, FMNH 83306; 

Tambopata Reserve, Explorer’s Inn, 30 km (airline) SSW 

Puerto Maldonado, 280 m, 12°50S, 69°17W, USNM 

222277–222280, 247337–247355, 247634–247644, 

268961–268967, 342998, 343243; Tres Chimbadas, ~20 

min downstream from Explorer’s Inn, 12°47S, 69°17 W, 

USNM 222275–222276, 222281. Ucayali: Balta, Río 

FIGURE 14. Distribution of Leptodactylus bolivianus (circles), L. guianensis (squares), and L. insularum (triangles). Question marks indicate 

localities where additional data are required to confirm the species identities.
number 635 • 3 5

Curanja, ~300 m, 10°8S, 71°13W, KU 196540–196554; 

Iparía, TNHC 37407; Pucallpa, 150 m, 8°23S, 74°32W, 

FMNH 56306–56307, 56446; Yarinacocha, 100–170 m, 

8°15S, 74°43W, FMNH 45402–45405, 56292, 56295–

56298, 56301.

VENEZUELA. Amazonas: [Río Casiquiare, 8 miles 

below Orinoco, 3°5N, 65°53W, AMNH 23167]; 

[Río Mavaca, 108 km SSE Esmeralda, 140 m, 2°15N, 

65°17W, USNM 216705–216723]; [Río Maturacá, Catarata de Huá (= Salto de Huá), 100 m, 0°48N, 66°21W, 

USNM 83575].

Leptodactylus guianensis, new species

Figure 15

Holotype. USNM 531509, an adult male 

from Guyana; Rupunini, Iwokrama Forest Reserve, Sipuruni River, Pakatau Camp, 4°4517’N, 59°0128’W, 

85 m. Collected by Maureen A. Donnelly and Megan 

Chen on 15 July 1997.

Paratopotyes. USNM 531508 (juvenile female), 531510–531511 (adult females), same locality and 

collectors; collected on 15 July 1997, 16 July 1997, and 

21 July 1997 respectively. 

Paratypes. AMNH 145130 and 145131 

(adult males, tissue sampled), Guyana: East Berbice–

Corontyne Region, Dubulay Ranch on the Berbice River, 

60 m, 5°4055N, 57°5132W; collected by Charles J. 

Cole and Carol R. Townsend on 30 August 1995.

All other specimens examined are referred specimens, 

not paratypes.

Etymology. The species is named for its distribution coinciding in large part with the Guiana Shield.

Diagnosis. Most specimens of L. guianensis

have boldly mottled posterior thighs with large light irregular spots/vermiculations on a dark background to the 

light marks being more extensive than the dark areas, the 

light markings sometimes coalesced; most specimens of 

L. bolivianus have the posterior thigh with a uniform or 

variable dispersion of melanophores (weakly to moderately 

mottled) usually for the entire extent of the posterior thigh. 

The only consistent character diagnosing the three species 

is the adult male thumb spine. The single spine on each 

thumb in L. guianensis is modestly chisel shaped (Figure 

13C), the single thumb spine of L. bolivianus is extremely 

chisel shaped (Figure 13B), and there are two rounded and 

pointed spines on each thumb in L. insularum (Figure 13A).

Description of Holotype. Figure 16. 

Snout rounded in dorsal and profile views. Canthus rostralis indistinct. Loreal concave-obtuse. Tympanum distinct, 

greatest diameter about 9/10 eye diameter. Tympanic annulus and adjacent area with numerous small keratinized 

tubercles. Vomerine teeth in two strongly arched series, 

narrowly separated from each other medially, positioned 

posteriorly to choanae. Each elliptical choana with anterior shelf extending over anterior edge of choana. Vocal 

slits elongate, parallel to jaw. Vocal sac not visible externally. Finger lengths in increasing order V < III < II < IV. 

Fingers with lateral fleshy ridges, clusters of small keratinized tubercles on upper side of finger II joints and rows 

of similar tubercles on the inner ridges of fingers III and 

IV. Finger subarticular tubercles pungent. Inner palmar tubercle large, oblong, pungent; outer palmar tubercle large, 

flat, elliptical, bifid distally, narrowly separated from 

inner tubercle. Smaller, rather indistinct palmar tubercles 

proximal to each subarticular tubercle and one indistinct 

palmar tubercle between metacarpal tubercles. Arms hypertrophied. Each thumb with one conical dark keratinized spine with a slightly developed chisel-shaped tip, tip 

on right thumb spine with three small cusps, tip on left 

with two small cusps. No large spines on chest. Upper and 

lower lips with numerous small keratinized tubercles. One 

central patch of numerous small keratinized tubercles on 

chest and a pair of tubercle patches on the lateral portions 

of the chest. Ulnar ridges weakly developed. Dorsal texture 

smooth anteriorly, with small scattered keratinized tubercles on posterior 2/3 of dorsum. A pair of well-developed 

dorsolateral folds extending from just posterior to the eye 

to the upper leg. A pair of slightly interrupted lateral folds 

from above the arm to groin. A pair of elongate glands 

above arm insertion. Belly disk fold distinct posteriorly 

FIGURE 15. Color photo of Leptodactylus guianensis from Coeroeni Island, Suriname (courtesy of Marinus S. Hoogmoed).
3 6 • smithsonian contributions to zoology

only. Scattered keratinized tubercles ventrally in arm insertion area. Toe tips rounded. Toes with well-developed 

fringes and basal webbing. Subarticular tubercles moderately pungent. Weak metatarsal ridge. Outer metatarsal 

tubercle almost rounded, about 1/3 size of prominent, 

elongately oval inner metatarsal tubercle. Tarsal fold well 

developed, straight except for most proximal end, extending about 9/10 length of tarsus. Tarsal fold with scattered 

small keratinized tubercles. Tarsal fold abutting inner 

metatarsal tubercle. Upper shank surface shagreened with 

scattered small keratinized tubercles. Scattered keratinized 

tubercles in arm insertion area. Outer tarsus and sole of 

foot with scattered small keratinized tubercles.

Upper lip with light outlined dark triangular mark, 

apex ending just short of eye. Light stripe from below eye 

becoming very distinct under the tympanum and extending 

to end of commissural gland. Dark stripe from tip of snout 

through nostril to anterior edge of eye. Dark posteriorly 

directed interorbital triangle continuous with chevron in 

scapular region, rest of dorsum nondescript. Pair of dark 

oval spots above arm insertion. Posterior portion of lateral 

fold distinctly lighter, some lateral warts light highlighted 

as well. Chin and anterior chest darkly mottled. Central 

chest and anterior belly with light and dark labyrinthine 

pattern, grading posteriorly to almost completely lacking 

melanophores except for a few small scattered dots. Posterior thigh mottled, overall dark appearance, with sizeable 

irregular lighter vermiculations.

Measurements (mm): SVL 100.5, head length 34.4, 

head width 35.5, eye–midnostril distance 10.4, greatest 

tympanum diameter (including annulus) 7.9, thigh length 

47.2, shank length 50.8, foot length 51.8.

Character Summary. The most frequent 

lip stripe patterns consist of a noticeable broad light 

A 

B 

C 

FIGURE 16. Dorsal view of (A) head, lateral view of head (B), and ventral view of the head, hands, and chest (C) of Leptodactylus

guianensis, new species, USNM 531509.

A 

B 

A C 

B 

C 
number 635 • 3 7

stripe extending from the front of the snout to the commissural gland, bordered below by continuous or broken 

markings on the upper lip and either bordered above by 

a dark line under the eye or not; or lip region uniform 

(about same intensity as background dorsal color) with 

or without a dark stripe under the eye and/or dark bars/

mottling on the upper lip. The most frequent posterior 

thigh pattern is boldly mottled with large light irregular 

spots/vermiculations on a dark background to the light 

marks being more extensive than the dark areas, the light 

markings sometimes coalesced. The most frequent lateral 

fold conditions are distinct folds only in the groin region, 

or indistinct overall, or indiscernible. The most frequent 

adult male chin tubercle condition is absence of tubercles.

Female (n = 52) SVL 66.0–109.2 mm (m [mean] = 

88.2), male (n = 34) 79.5–109.5 mm (m = 94.8). Female 

head length/SVL ratio 0.33–0.40 (m = 0.359), male 0.33–

0.39 (m = 0.365). Female head width/SVL ratio 0.31–0.37 

(m = 0.327), male 0.32–0.39 (m = 0.352). Female eye–

nostril distance/SVL ratio 0.09–0.11 (m = 0.099), male 

0.09–0.12 (m = 0.101). Female tympanum diameter/SVL 

ratio 0.06–0.08 (m = 0.069), male 0.06–0.08 (m = 0.072). 

Female thigh length/SVL 0.42–0.52 (m = 0.476), male 

0.44–0.53 (m = 0.484). Female shank length/SVL ratio 

0.48–0.57 (m = 0.529), male 0.50–0.56 (m = 0.525). Female foot length/SVL ratio 0.48–0.58 (m = 0.527), male 

0.46–0.57 (m = 0.527).

Tadpole. Unknown.

Advertisement Call. Unknown.

Distribution. Lowland portions of Guiana 

Shield regions of Guyana, Suriname, Venezuela, and adjacent Brazil (Figure 14).

Specimens Examined (n = 226). Localities enclosed by brackets require validation that the species is correctly identified. BRAZIL. Roraima: Boa Vista, 

90 m, 2°49N, 60°40W, MZUSP 62482, 65508–65510, 

66053; BR-174 at km 110 (Rio Surumu), 3°25N, 

60°18W, MZUSP 60616; Marco da Fronteira BV-8, 

4°28N, 61°8W, INPA 1306, MZUSP 65760–65761, 

65783–65786, USNM 302445; [Cachoeira do Cujubim, Rio Catrimani, 1°45N, 62°17W, MZUSP 58533]; 

Cachoeira do Paredão, 2°57N, 61°35W, MZUSP 70634; 

Colonia Apiaú, 150 m, 2°38N, 61°12W, MZUSP 65923, 

65948, 65953, 66317–66319, 68632–68633, USNM 

302268–302270; Fazenda Salvamento, 3°20N, 61°24W, 

MZUSP 66031; Igarapé Cocal, near Tepequém, 3°45N, 

61°44W, MZUSP 66008, 67004; Ilha do Maracá, 3°25N, 

61°40W, MZUSP 60628, 62420–62426, 65548–65558, 

65573, 65613, 65670–65672, 65698–65700, 65703–

65705, 65735–65736, 65743–65744, 70649–70650, 

70724; [Missão Catrimani, 0°27N, 61°41W, MZUSP 

68734–68736]; [Parque Nacional Viruá, Roraima, próximo a Caracaraí, Igarapé Viruá, 1°0N, 61°15W, AL 

12241–12242]; [Praia do Guariúba, 0°20N, 61°44W, 

MZUSP 67237–67240, USNM 302471–302473]; [Santa 

Maria da Boiaçu, MZUSP 67330, 67332–67335, 68296]; 

[Serra do Surucucus, 2°47N, 63°40W, MZUSP 65817]; 

[Uiaca, Rio Uraricoera (= Uaicás, Rio Urariquera), 3°33N, 

63°11W, AMNH 100009–100010].

GUYANA. Region unknown: Anowini Creek, Essequibo River, UMMZ 79474; Essequibo River, AMNH 

49257. Barima–Waini Region: Maburama, 100 m, 

8°12N, 59°47W, UTA 55504. Cuyuni–Mazaruni Region: Blackwater Creek, 6°5N, 59°59W, AMNH 

43670; Bartica, 3 m, 6°24N, 58°37W, MCZ-A 50704; 

Kalacoon, Mayarum River (= Mazaruni River), AMNH 

3990; Kartabo, 100 m, 6°21N, 58°41W, AMNH 10397, 

10423, 11709; Paruima (= Paruima Mission, Kamarang River), 614 m, 5°48N, 61°1W, UMMZ 85151, 

58152(2). Demerara–Mahaica Region: Georgetown, sea 

level, 6°48N, 58°10W, AMNH 5064. East Berbice–

Corontyne Region: Dubulay Ranch on the Berbice River, 

60 m, 5°4055N, 57°5132W, AMNH 145130–145131, 

145133–145134. Potaro–Siparuni Region: Magdalen’s 

Creek Camp, near (~300 yards) [275 m] NW bank of 

Konawaruk River (~25 miles [40 km] linear) WSW Mabura Hill, 120 m, 5°137N, 59°243W, AMNH 166359–

166366. Upper Demerara–Berbice Region: Berbice River 

camp at ~18 miles [30 km] (linear) SW Kwakwani (~2 

miles [3.2 km] downriver from Kurundini River confluence), 60 m, 5°56N, 58°1414W, AMNH 166341–

166349, 166356–166358; Malali (= Malali rapids), 100 

m, 5°37N, 58°22W, AMNH 45773; Penal Settlement, 

6°24N, 58°39W, AMNH 10422; Rockstone, 100 m, 

5°59N, 58°33W, FMNH 26633–26636, UMMZ 80442; 

Wismar, ~100 m, 6°0N, 58°18W, UMMZ 76681(2). 

Upper Takutu–Upper Essequibo Region: North of Acarahy 

Mountains, west of New River, 1°50N, 57°30W, KU 

69667–69686; Iwokrama Forest Preserve, Burro-Burro 

river, Burro-Burro Camp, 83 m, 4°4352N, 58°512W, 

AMNH 164217–164218, USNM 531502; Iwokrama Forest Preserve, Kabocalli Camp, 101 m, AMNH 164219–

164221; Iwokrama Forest Preserve, ~3 miles [4.8 km]

S Kurupukari Base Camp on Georgetown-Lethem road, 

Three Mile Camp, 102 m, 4°3759N, 58°4252W, 

USNM 531501; Iwokrama Forest Preserve, 5 hours 

(downstream) Kurupukari Base Camp on Essequibo 

River, Kabocali Camp, 101 m, 4°176N, 58°3034W, 

USNM 531503–531507; Iwokrama Forest Preserve, Sipuruni River, Pakatau Camp, 85 m, 4°4517N, 59°128W, 
3 8 • smithsonian contributions to zoology

USNM 531508 (Paratype, juvenile female), 531509 

(Holotype, male), 531510–531511 (Paratypes, females); 

Iwokrama Forest Preserve, Pakatau Creek, 85 m, 4°45N, 

59°1W, AMNH 164223–164227; northern Rupununi Savanna, Karanambo (on Rupununi River), McTurk Ranch, 

110 m, AMNH 136033–136034; Karanambo Ranch, 

Maricuba Lake, ~100 m, 3°458N, 59°1836W, USNM 

497730; Kuyuwini, 1°55N, 59°15W, AMNH 43439, 

43684, 46278–46279, 46286; Moco-Moco (= Mocomoco), 29 km SE Lethem, base of Kanuku Mountains, 

100 m, 3°1812N, 59°390W, USNM 497731; Mocho 

Mocho (= Moco Moco, = Mocomoco), USNM 146369–

146374; Onora River (= Unorowo River), tributary of Essequibo River, 1°35N, 58°30W, AMNH 49331–49335, 

53440–53445.

SURINAME. Brokopondo: Brownsberg Nature Park, 

near Mazaroni top, ~450 m, 4°55N, 55°12W, AMNH 

87757; Loksietattie, Saramacca River, 5°2N, 55°32W, 

FMNH 134739. Commewijne: Plantation Ma Retraite, 

sea level, 5°52N, 55°8W, AMNH 125820. Nickerie:

Kaiserberg airstrip, Zuid River, FMNH 128810–128818, 

128820–128825, 128919, 128926, 128929, 128944. 

Para: near Zanderij, sea level, 5°27N, 55°12W (coordinates for Zanderij), USNM 159513–159514. Saramacca:

Foengoe Island Airstrip, Raleigh cataracts, Coppename 

River, 50 m, 4°44N, 56°12W, AMNH 87758. Sipaliwini: 

Drietabbetje, Tapahony River, 4°7N, 54°40W, MCZ-A 

97282; Marapi LLB Station on Covantyn River, 5°N, 

57°W, MCZ-A 110622. Wanica: Pad (Path) van Wanica, 

5°47N, 55°20W, KU 204508.

VENEZUELA. Bolívar: Arabupo (= Arabopo), 

1,200–1,300 m, 5°6N, 60°44W, UMMZ 85198; ~35 km 

S El Manteco, 350 m, 5°6N, 62°37S, TCWC 60158; 13 

km S, 1 km E Puente Cuyuni, 140 m, 6°43N, 61°35W 

(coordinates for Puente Cuyuni), KU 166405–166408; 

Santa Lucia de Surukun, 45 km NE Icabaru, 851 m, 

4°33N, 61°25W, USNM 216726.

Leptodactylus insularum Barbour, 1906

Figure 17

Leptodactylus insularum Barbour, 1906:228. Type locality: Saboga Island, 

Panama.

Barbour (1906) described Leptodactylus insularum sp. 

nov. based on a lot of 12 specimens with the single MCZ 

number 2424 from Saboga Island in the Gulf of Panama. 

The type description is not based on a single specimen 

but is a composite description including male and female 

data. Subsequently, 11 of the 12 original syntypes were 

exchanged or had new MCZ catalogue numbers assigned 

to them. There remains one specimen with MCZ number 

2424. As the specimen bearing number 2424 is the only 

specimen currently with that number in the MCZ collection, we hereby designate MCZ 2424 as the lectotype of 

Leptodactylus insularum Barbour, 1906.

Lectotype Description. Adult female 

(Figure 18) with pigmented ova. Snout nearly rounded 

in dorsal view, rounded in profile view. Canthus rostralis indistinct. Loreal concave-obtuse. Tympanum distinct, 

greatest diameter about 7/10 eye diameter. Tympanic 

annulus smooth. Vomerine teeth in arched series, apices 

anterior, narrowly separated from each other medially, 

posterior to choanae. Choanae damaged, shape uncertain. 

Finger lengths II = IV > III ≈ V. Fingers with weak lateral 

ridges or smooth. Finger subarticular tubercles rounded, 

not pungent. Inner palmar tubercle large, ovate, prominent; outer palmar tubercle present, somewhat indistinct, 

right hand tubercle weakly heart-shaped, left present, 

shape unclear. Dorsum smooth. A pair of distinct dark 

dorsal folds from just behind eye to sacrum, folds indistinct from sacrum to juncture of leg and body. Supratympanic fold dark brown from eye to behind jaw. Short dark 

lateral folds extending from supratympanic fold at level 

of posterior tympanum to ending shortly after posterior 

arm insertion. No distinct glands. Belly disk fold indistinguishable. Arms and body lacking tubercles. Toe tips 

round, swollen, just larger than toes immediately behind 

tips. Toes with lateral fringes and weak basal webbing between toes I–IV. Toe subarticular tubercles large, rounded. 

FIGURE 17. Photo of Leptodactylus insularum from San Andrés, 

Colombia (courtesy of Roy W. McDiarmid).
number 635 • 3 9

Weak outer metatarsal ridge continuous with outer ridge 

on toe V. Outer round metatarsal tubercle about 1⁄4 size of 

oval inner metatarsal tubercle. Distinct tarsal fold lacking tubercles, extending from and continuous with inner 

metatarsal tubercle to about 9/10 length of tarsus. Upper 

thigh surfaces smooth. Upper shank surfaces with numerous small white tubercles. Outer surface of tarsus with 

small white tubercles, a few of which with brown keratinized tips. Sole of foot smooth. 

Face with indistinct markings, a slightly darker indistinct interrupted stripe from nostril to eye, postocular area 

darker brown than upper lip area. Dorsum brown with 

indistinct darker marks scattered between dorsolateral 

folds. Flanks with irregular dark brown spots on tan to 

brown background. Belly with small tan dots visible under 

magnification, overall belly very light tan, throat slightly 

darker. Posterior thigh surfaces with a pattern of large 

darker and lighter brown mottling.

Measurements (mm): SVL 83.8, head length 29.2, 

head width 29.1, eye–nostril distance 7.8, tympanum diameter (including annulus) 5.6, thigh length 38.8, shank 

length 42.3, foot length 24.6.

Etymology. Barbour did not specifically indicate the derivation of the species name. All of the specimens indicated in the original description are from islands 

in the Gulf of Panama, hence the specific name. 

Diagnosis. Most specimens of L. insularum

have boldly mottled posterior thighs with large light irregular spots/vermiculations on a dark background to 

the light marks being more extensive than the dark areas, 

the light markings sometimes coalesced; most specimens 

of L. bolivianus have the posterior thigh with a uniform 

or variable dispersion of melanophores (weakly to moderately mottled) usually for the entire extent of the thigh. 

Leptodactylus insularum adult males only have central 

chest patches of tubercles; L. bolivianus males have lateral 

FIGURE 18. Dorsal and ventral views of the lectotype of Leptodactylus insularum, MCZ 2424.
4 0 • smithsonian contributions to zoology

patches of chest tubercles in addition to the central patch. 

The one entirely consistent character diagnosing the three 

species is the adult male thumb spine. The two spines on 

each thumb of L. insularum are rounded and pointed (Figure 13A); the single spine on each thumb of L. bolivianus

and L. guianensis is modestly to extensively chisel shaped 

(Figure 13B, C).

Character Summary. The most frequent 

lip stripe pattern consists of lip region uniform (about 

same intensity as background dorsal color) with or without a dark stripe under the eye and/or dark bars/mottling on the upper lip. The most frequent posterior thigh 

pattern is boldly mottled with large light irregular spots/

vermiculations on a dark background to the light marks 

being more extensive than the dark areas, the light markings sometimes coalesced. The most frequent lateral fold 

condition is fold distinct or slightly interrupted from supratympanic fold to leg. The most frequent adult male 

chin tubercle condition is presence of tubercles.

Female (n = 224) SVL 60.4–99.1 mm (m [mean] = 

81.3), male (n = 216) 66.0–104.6 mm (m = 86.8). Female 

head length/SVL ratio 0.33–0.43 (m = 0.365), male 0.32–

0.41 (m = 0.364). Female head width/SVL ratio 0.30–0.38 

(m = 0.334), male 0.32–0.40 (m = 0.353). Female eye–

nostril distance/SVL ratio 0.08–0.12 (m = 0.102), male 

0.09–0.12 (m = 0.102). Female tympanum diameter/SVL 

ratio 0.05–0.08 (m = 0.070), male 0.06–0.08 (m = 0.070). 

Female thigh length/SVL ratio 0.36–0.58 (m = 0.452), 

male 0.39–0.51 (m = 0.451). Female shank length/SVL 

ratio 0.44–0.58 (m = 0.495), male 0.43–0.58 (m = 0.489). 

Female foot length/SVL ratio 0.44–0.57 (m = 0.506), male 

0.42–0.59 (m = 0.503).

Tadpole. Figures 19, 20. Exotrophic, lentic, 

benthic guild member (McDiarmid and Altig, 1999, guild 

IIA1); oral disk anteroventrally positioned, entire (not 

emarginated), moderate anterior papilla gap (~31% oral 

disc width), single and/or double row of marginal papillae; tooth row formula 2/3; spiracle sinistral; vent median; 

dorsal fin origin at tail-body junction; dorsum with a relatively uniform profusion of melanophores, venter with 

light to moderate profusion of melanophores, venter with 

light to moderate profusion of melanophores from oral 

disk to guts, scattered melanophores over guts, tail musculature uniform moderate to dark brown, tail fins ranging 

FIGURE 20. Illustration of oral disk of Leptodactylus insularum

tadpole, USNM 576264, Costa Rica, Guanacaste, ~12 km (airline) 

SSW Cañas, scale bar = 1 mm.

FIGURE 19. Illustration of lateral view of Leptodactylus insularum tadpole, USNM 576264, Costa Rica, Guanacaste, ~12 km (airline) 

SSW Cañas, scale bar = 5 mm.
number 635 • 4 1

from same pattern as tail musculature to ventral fin with 

a gradient of no melanophores next to body to uniform 

brown around midfin; maximum total length stage 37, 

35.2 mm (based on voucher specimens KU 68365, USNM 

241340, 330406, 330837, 576264).

This description overall agrees with the previous one 

reported by Heyer (1970). 

Advertisement Call. Calls of single notes 

at rates of 1.2–2.5/s; call duration 0.08–0.12 s; call amplitude modulated, loudest portion from about midcall to 

4/5 call duration; call frequency modulated beginning at 

110–220 Hz with the highest frequency starting about at 

1/3 call length of 890–1,200 Hz (Figures 6, 7) (based on 

voucher recordings from Fouquette, 1960; Ibáñez et al., 

1999; and KU recording 801).

Distribution. Mainland Pacific versant of 

Costa Rica, throughout lowland Panama, Caribbean drainages of Colombia and Venezuela, Trinidad (Figure 14).

Specimens Examined (n = 1,018). COLOMBIA. Antioquia: Chigorodó, ~100 m, 7°41N, 

76°42W, AMNH 76186, USNM 151887; El Cinco, 

7°4535.4”N, 74°4649.1W, MPUJ 2073–2074; Jericó, 

1967 m, 5°47N, 75°47W, MCZ-A 24886; Medellin, 

1500 m, 6°15N, 75°35W, AMNH 38828; San Luis, corregimiento La Danta, near Puerto Carreño, ICNMHN 

17225; Nechí, 100 m, 8°7N, 74°46W, FMNH 54568, 

54571; 20 km abajo del [below the] Río Nechí, frente 

al caserio [in front of the village] Bijagual, 7°5013N, 

74°4743.7W, MPUJ 2840; Urabá, Río Currulao, 50 m, 

7°0N, 75°30W, FMNH 63847. Arauca: Estación Cravo 

Norte region, ICNMHN 26833–26841, 27226–27231, 

27680–27684, 27890. Atlántico: Barranquilla, Airport 

Soledad, sea level, 10°55N, 74°46W, USNM 152641; 

Municipio Los Pendales, 10°37N, 75°13W, ICNMHN 

42275; Estación Piscicultura de Soplaviento, 10°2336N, 

75°828W, IND-AN 567, 569, 572, 576, 579–581; 

Puerto Colombia, 10°5928N, 74°5743W, ICNMHN 

1020; Santa Lucia, 10°21N, 74°58W, ICNMHN 46136; 

Tubará, corregimiento Cuatrobocas, 10°55N, 73°57W, 

ICNMHN 49010. Bolívar: no other locality data, MPUJ 

2187–2190; Isla Fuerte, 9°25N, 76°10W, FMNH 

152371; San Cristóbal, Canal del Dique, ~100 m, 9°53N, 

75°15W (coordinates for San Cristóbal), AMNH 75742–

75746, 75748; Hacienda El Ceibal, 80 m, 10°37N, 

75°14W, ICNMHN 44117–44135; Hacienda La 

Aguada, 22 km from San Onofre, USNM 152653; Santa 

Rosa, 600 m, 7°58N, 74°3W, ICNMHN 2102, 2105–

2106, 2108, 2245–2246, 2262–2287, 2289. Boyacá: Las 

Mercedes, Hacienda Los Balcones, 380 m, ICNMHN 

38433–38437; Muzo, 1240 m, 5°33N, 74°6W, IND-AN 

2739, 2742; Puerto Boyacá-Puerto Romero carretera, km 

29–35, 300–320 m, ICNMHN 38438–38440; Puerto 

Boyacá, Insp. Puerto Romero, ICNMHN 44726–44730; 

Puerto Boyacá, Vereda El Okal, Quebrada La Fiebie, km 

29–30 Otanche road, 280 m, ICNMHN 45427–45437, 

45433, 45438. Caldas: Finca Tintiná, 5°1429.5N, 

75°4111.1W, MPUJ 1827; Guarinocito, 280 m, 5°23N, 

74°43W, AMNH 81855–81858, CMB 548–549, MPUJ 

374; Reserva Riomanso, 5°40N, 74°46W, MPUJ 3016; 

Sitio La Turqueza, 5°19.9N, 75°425.5W, MPUJ 

1825–1826, 2075; 15.6 km carretera de la Victoria a la 

Central, ICNMHN 34607–34608. Casanare: Orocué, 

200 m, 4°48N, 71°20W, ICNMHN 34683; San Luis 

de Palenque, 5°25N, 71°40W, ICNMHN 2664. César:

Finca El Diamante, 15 km south of Bosconia, 9°58N, 

73°51W (coordinates for Bosconia), USNM 200350–

200353; Casacara, Municipio Codazzi, 9°20N, 74°45W, 

ICNMHN 39935–39937; Finca Mundo Nuevo, 9°45N, 

73°33W, MPUJ 1388–1390; Valle Dupar (= Valledupar), Santa Marta mountains, 202 m, 10°29N, 73°15W, 

UMMZ 54597–54598. Chocó: Acandí, corregimento Gilgal, 2000 m, ICNMHN 18112–18114; Parque Nacional 

Natural Katios, 7°50N, 77°7W, ICNMHN 3224, 47167–

47174, 47177–47186, IND-AN 2000–2002; Río Sucio 

Hacienda Sautatá, 20 m, 7°500N, 77°45W, ICNMHN 

187–192; Río Sucio, Vereda Las Teresitas, 7°925N, 

77°2815W, ICNMHN 47188. Córdoba: Ayapel, casco 

Urbano, 8°1858N, 75°830W, ICNMHN 48165; 

Ayapel, corregimiento Nariño, caserío Playa Blanca, 

8°1747N, 74°5846W, ICNMHN 48166; Ayapel, Hacienda La Balsa, 8°185N, 75°224W, ICNMHN 48167; 

Ayapel, Hacienda Quibrache, 8°1747N, 74°5846W, 

ICNMHN 48163; Lórica, Bosque Ita, 20 m, 9°16N, 

75°49W, ICNMHN 48172; Lórica, Estación Piscicola, 

CVS, 20 m, 9°13N, 75°50W, ICNMHN 48168–48170, 

48320; Lórica, La Montaña del Mono, 20 m, 9°13N, 

75°50W, ICNMHN 48171; Monte Libano (= Montelíbano), 8°5N, 75°29W, ICNMHN 42755–42756, 

MZUSP 106090–106091; Montería, Hacienda El Diluvio, 64 m, 8°4318N, 75°5926W, ICNMHN 48159–

48161; Municipio de Montería, ICNMHN 19550–19551; 

Montería, Maracayo, 8°2432, 75°5326W, ICNMHN 

48157–48158; Montería, Quebrada Betancí, 8°2244N, 

75°4955W, ICNMHN 48156; Pueblonuevo, Hacienda 

Toronto, 8°24N, 75°17W, ICNMHN 48173; Represa de 

Urrá, 70–150 m, 8°14N, 76°1253W, ICNMHN 41313; 

Río Manso, tributary upper Río Sinu, 7°41N, 76°10W, 

USNM 151031–151033; Tierra Alta, 70 m, 8°11N, 

76°4W, FMNH 61807–61812, KU 144967–144977, 

UMMZ 132466(4). Cundinamarca: Beltrán, 230 m, 
4 2 • smithsonian contributions to zoology

4°47N, 74°47W, ICNMHN 2339–2343, 2603, USNM 

145708–145742, 147254; Girardot, ~300 m, 4°18N, 

74°48W, AMNH 71575–71577, 75158, USNM 146216–

146218, 146393, 146467; Girardot, Isla El Sol, al Este 

de la confluencia del Río Bogotá, 310–320 m, ICNMHN 

273–275, 277–279, 281–293. Guajira: Municipio Patomino, 11°148.1N, 73°337.3W, MPUJ 2897; Nazareth 

(= Nazaret), 200 m, 12°11N, 71°17W, USNM 115112–

115114, 194735; near Piojo, USNM 152676–152677; 

Río Barbacoas, Arroyo de Arenas, 11°25N, 73°5W, 

UMMZ 54601; Riohacha, corregimiento Calabacito, 

130 m, 11°33N, 72°55W (coordinates for Riohacha), 

ICNMHN 11859–11865. Huila: Campoalegre, 530 m, 

ICNMHN 9365; Finca La Tribuna, 3°44N, 75°223W, 

MPUJ 3573–3576; Finca San Rafael, 3°315N, 

75°625.5W, MPUJ 1786–1787; Parque Nacional Natural Cueva de los Guacharos, Municipio Acevedo, 1200 m, 

1°35N, 76°1W, USNM 148859. Magdalena: Balneario 

El Rodadero de Guaira, 11°12N, 74°13W, KU 169075–

169089; Bolívar, Santa Marta mountains, 10°21N, 

74°9W, UMMZ 54596; Ciénaga, sea level, 11°1N, 

74°15W, USNM 144157–144578; Don Diego, sea level, 

11°15N, 73°42W, UMMZ 48118, 48119(3), 48488(10); 

Fundación, 60 m, 10°26N, 74°7W, MCZ-A 8966–8967, 

UMMZ 48106–48108, 48110–48111, 48113–48117; 

Isla Salamanca–Los Cocos, sea level, 10°59N, 74°27W, 

IND-AN 622–626, 6629; Mamatoco (= Manatoca, Mamatoca), 11°15N, 74°9W, UMMZ 48120; Minca, on 

road to San Lorenzo, 11°12N, 74°4W, USNM 150926; 

Parque Nacional Natural Tayrona (= Tairona, Tayruna), 

IND-AN 2462, 2523; Parque Nacional Natural Tayrona, 

Arrecifes, 11°1928.5N, 73°5743.7W, IND-AN 2483; 

Parque Nacional Natural Tayrona, Cañaveral, sea level, 

11°19N, 73°56W, IND-AN 2441–2442, 2488–2490; 

Parque Nacional Natural Tayrona, 2–3 km cabaña Cañaverales, ICNMHN 13611–13613, 13659; Parque 

Nacional Natural Tayrona, Cinto, 11°20N, 74°4W, 

IND-AN 2443, 2446; Parque Nacional Natural Tayrona, 

trail between Cañaveral and El Pueblito, USNM 200348–

200349; Parque Nacional Natural Tayrona, El Cedro, 

11°1954.67N, 74°054.31W, IND-AN 2463; Parque 

Nacional Natural Tayrona, within 2 km of El Cedro station, USNM 200354; Parque Nacional Natural Tayrona, 

near La Gayraca, USNM 200356; Parque Nacional Natural Tayrona, Los Naranjos, 150 m, 11°18N, 73°54W, 

IND-AN 2482; Río Buritaca, 11°16N, 73°47W (coordinates for mouth of river at Atlantic Ocean), USNM 

150992–150994; Río Frio, 30 m, 10°55N, 74°10W, 

MCZ-A 16067–16068; between Santa Marta and Mamatoco (Manatoca, Mamatoca), USNM 150893–150924. 

San Andrés y Providencia: Isla de Providencia, 13°21N, 

81°22W, MCZ-A 19261–19266, USNM 76956–76957, 

146214–146215; Isla de Providencia, Agua Mansa, 

IND-AN 1096–1102; Isla de Providencia, reservoir, 80 m, 

ICNMHN 33421–33427; Isla de San Andrés, 12°32N, 

81°42W, AMNH 71578–71580, KU 265811–265812, 

MCZ-A 17370–17375; Isla San Andrés, near Punta 

Paraiso, UMMZ 127882. Santander: El Centro, 100–

200 m, 6°57N, 73°46W, FMNH 81759, 81761, USNM 

144832–144837. Sucre: Hacienda La Estanzuela, 4 km 

east of Tolú, 9°31N, 75°32W, USNM 200343–200347; 

Municipio San Juan de Onofre, corregimiento de Berrugas, Finca Los Morros, 5 m, ICNMHN 43006; Sincelejo 

to Tolú, USNM 150743–150749. Tolima: El Cardonal, 

5°513N, 74°4153W, MPUJ 1402, 1405, 1409, 1416, 

1418–1419, 1421; Espinal, Magdalena Valley, 200 m, 

4°7N, 74°53W, MCZ-A 15067, 15071; Finca El Antonio, 3°4737.8N, 75°187W, MPUJ 1788; Guayabal, 

4°58N, 74°54W, MPUJ 1391–1394, 1398; Honda, 

230 m, 5°12N, 74°45W, AMNH 22604, UMMZ 76073; 

4 km northwest of Honda, 5°14N, 74°46W, AMNH 

57549–75751; Municipio Margarita, ICNMHN 42765–

42766; Melgar, orillas Río Sumapáz, 400 m, 4°12N, 

74°39W, ICNMHN 1422, 18143; Municipio Venadillo, 

4°4317N, 74°5554W, ICNMHN 43172.

COSTA RICA. Guanacaste: Estación Experimental Enrique Jiménez Nunez, 13.6 miles SW Cañas, 20 m, 

10°20N, 85°9W, USNM 219762–219763; Río Bebedero, 5 km S Bebedero, 5 m, KU 65653–65656. Puntarenas: Dominical, ~6.4 km N, 9°17N, 83°51W, KU 

204367; Golfito, 10 m, 8°38N, 83°11W, KU 32294–

32304, 65651–65652, 86292; Palmar (= Palmar Sur), 

8°58N, 83°27W, KU 32293, 65648; Palmar Sur, 5.6 km 

SE, 8°56N, 83°25W, KU 65646; Palmar Sur, 7 km SE, 

8°56N, 83°25W, KU 65647; Parrita, 9°30N, 84°19W, 

MZUSP 106680; Rincón de Osa, 8°42N, 83°29W, KU 

102098, USNM 227621–227624; San Isidro del General, 

15–20 miles [24–32 km] W on Dominical road, 9°16N, 

83°50W, KU 32392; Villa Neilly, 1.6 km WNW, 75 m, 

8°40N, 82°55W, KU 65649–65650; Volcan Buenos 

Aires, 1 mile [1.6 km] E, Cone Finca, 9°13N, 83°25W, 

UMMZ 117577.

PANAMA. Province unknown: Between Campana and 

La Venta, USNM 103481; Canal Zone, KU 67947; Istmo 

do Panamá, MZUSP 5454; Río Morte Arnade,USNM 

53928; Tabernilla, FMNH 16683. Chiriquí: David, 0.7 

miles [1.1 km] north, 0.2 miles [0.3 km] west, 8°26N, 

82°26W, AMNH 69714; Progreso, 8°27N, 82°50W, 

UMMZ 58153–58165; Puerto Armuelles, 8°17N, 

82°52W, MCZ-A 24236–24238; Puerto Armuelles, 7.5 
number 635 • 4 3

km N, 8°19N, 82°49W, KU 107163–107197. Coclé:

Penonomé, 6 km SSW, 30–70 m, 8°31N, 80°22W, KU 

107203; Río Chorrera, 8°18N, 80°23W, USNM 53885; 

Valle de Anton, 560 m, 8°37N, 80°8W, USNM 139703. 

Colón: Achiote, 9°1322N, 80°19W, KU 76514–76516; 

France Field, 9°22N, 79°52W, MCZ-A 10143–10144; 

Frijoles, lake at, 9°10N, 79°50W, FMNH 6003–6004; 

Gatun, 9°16N, 79°55W, FMNH 16658. Darién: Camp 

Townsend, Camp Creek, 8°10N, 77°42W, AMNH 

40931, 40998–40999, 41027–41031, 41033–41037, 

41063, 41727; Camp Townsend, below Yavisa, Río Chucunaque on Río Darién, 8°10N, 77°42W, AMNH 41715; 

Cana, 460 m, 7°4555N, 77°416W, USNM 50222; Canclones, 8°20N, 77°46W, UMMZ 132664, 132667(5), 

132668–132669; Canglon, camp site below, Río Chucunaque, AMNH 41128, 41130–41131; Chucunaque, 

Río Chucunaque, AMNH 41121–41122; Camp Chucunaque, ~0.5 miles [0.8 km] S, AMNH 40614–40615, 

40643–40644, 40699, 40713–40715, 40929, 41088–

41089, 41734, 41766, FMNH 170431, KU 80380–80384, 

107209–107222, UMMZ 125006, 137749–137750, 

USNM 140575–140583; Jaqué, 7°31N, 78°10W, KU 

115280; Mount Sapo, 7°5831N, 78°2143W, MCZ-A 

9179–9180; Ortiga Camp, 8°45N, 78°0W, FMNH 

170432, 170464; Río Abobi, Río Chucunaque, AMNH 

40897; mouth of Río Canglon, Río Chucunaque, 8°20N, 

77°46W, AMNH 40585–40587, UMMZ 123281(2), 

124998–125002, 125003(2), 125004–125005, 132670; 

Río Chico, near Avelinos, 8°12N, 77°36W, AMNH 

39787, 40979–40984; near Río Membrillo, Río Chucunaque, 8°39N, 77°47W, KU 116816–116821; ~7 km 

above Río Mortí, Río Chucunaque, 8°52N, 77°59W, KU 

107223–107227, 107906; just below mouth of Río Mortí, 

Río Chucunaque, 8°50N, 77°59W, UMMZ 132663; intersection of Río Chucunaque and Río Ucurganti, 8°25N, 

77°48W, USNM 140616–140619; Río Chucurti (= Subcurtí), near camp, 8°45N, 77°57W, AMNH 40622, 

40627; near mouth of Río Clarita, 8°6N, 77°28W, 

UMMZ 132665–132666; Río Jesusito, Sapo Mountains, 

8°1N, 78°16W, MCZ-A 9167; mouth of Río Membrillo, 

8°39N, 77°47W, AMNH 40551–40554, Río Pita, 10 m, 

FMNH 67884, 67886–67892; camp above Río Sansan (= 

Río Sansón), Río Chucunaque, 8°24N, 77°47W, AMNH 

40646–40647; Río Suchuti (= Río Sucutí = Río Subcurtí), 

8°45N, 77°57W, AMNH 40798; mouth of Río Tuquesa 

on Río Chucunaque, 8°23N, 77°47W, AMNH 40761–

40765; Río Ucurganti, ~7 km above mouth, 30 m, 8°28N, 

77°48W, KU 10725, 116822–116823; Santa Fe camp, 

FMNH 170434–170435; Three Falls Creek (= Camp 

Creek, Camp Townsend), 8°10N, 77°42W, AMNH 

41711; Yavisa, across stream from, 8°11N, 77°41W, 

AMNH 41145. Herrera: Parita, 8°0N, 80°31W, USNM 

127262–127286; 5 km northwest Santa María, near Río 

Santa María, 8°8N, 80°41W, KU 107202. Los Santos: 6 

km (road) southwest Los Santos, 7°56N, 80°25W (coordinates for Los Santos), USNM 203649; Tonosí, 40 m, 

7°24N, 80°27W, KU 108616. Panamá: Ancon, 8°58N, 

79°33W, MCZ-A 10142; Balboa, 8°57N, 79°34W, KU 

107204; Barro Colorado Island, 9°11N, 79°57W, FMNH 

173206, MCZ-A 19207; Briya Point near Palo Seco, near 

Balboa, 8°55N, 79°34W (coordinates for Palo Seco), 

MCZ-A 10145; Bruja Point, MCZ-A 10669, 11750; Cocoli, 8°59N, 79°35W, USNM 143347; Fort Clayton, 

9°0N, 79°45W, MCZ-A 10141; Fort Kobbe, 8°55N, 

79°35W, USNM 193348; Howard Air Force Base, 

8°55N, 79°36W, USNM 193337, 193339; Isla Chapera, 

10 m, 8°35N, 79°2W, AMNH 87337–87339; Isla Cantadora, 8°38N, 70°02W, AMNH 87334–87336; Isla Saboga, 10 m, 8°37N, 79°4W, AMNH 87340–87344; Juan 

Mina, 25–30 m, 9°10N, 79°39W, FMNH 57537–57539, 

60488, 67883, KU 107206, UMMZ 135375; Los Overos, 

on Río Indio, road to Las Minas, 593785 0988063 UTM, 

USNM 565027–565028; Miraflores, 9°0N, 79°36W, 

USNM 53841; Nueva Gorgona, 8°33N, 79°52W, AMNH 

69715–69719; Panama City (= Old Panama = Panama 

Viejo), 8°58N, 79°32W, AMNH 41068–41070, 52742, 

FMNH 60487, 170433, KU 116814–116815; Pipeline 

Road, 9°7N, 79°47W to 9°15N, 79°49W, UMMZ 

152934; Puerto La Chorrera, near, 8°52N, 79°43W; Río 

Mamone (= Río Mamoní), El Capitán (= La Capitana), 

9°9N, 79°4W, FMNH 16657, USNM 53982; Río Siluganti (= Río Chuluganti), 9°1313N, 78°489W, UMMZ 

137745–137748; Río Tocumen, 9°1N, 79°23W, MCZ-A 

10140; Rousseau, 8°58N, 79°35W, KU 67959; Saboga Island, 10 m, 8°37N, 79°4W, MCZ-A 2444, 99314–99319; 

San Miguel (= Isla San Miguel), 8°22N, 78°55W, AMNH 

6669, MCZ-A 2424, 6901–6902, UMMZ 48076, USNM 

58062; Summit Gardens, 9°4N, 79°39W, KU 107904–

107905, UMMZ 167314; Tapia, 9°4N, 79°25W, AMNH 

18915–18917, 18920; Tocumen, 9°5N, 79°23W, USNM 

147504; 8 km NE Tocumen, 9°6N, 79°20W, KU 107207–

107208; near Venado Beach, 8°54N, 79°36W, USNM 

19336. San Blas: Camp Sasardí, 12 m, 9°0N, 77°48W, 

KU 115275–115279. Veraguas: 3.4–5 km N Montijo, 

7°59N, 81°4W, KU 107907–107908, 107198–107201.

TRINIDAD. Nariva: Brickfield, 10°20N, 61°16W, 

FMNH 49661–49663. St. Patrick: Icacos Peninsula, 

USNM 306180; Icacos Point, 10°2N, 61°55W, USNM 

287010; Laurier Pond near Siparia, 10°8N, 61°30W (coordinates for Siparia), FMNH 218782–218783.
4 4 • smithsonian contributions to zoology

VENEZUELA. Anzoategui: Carapá, near, 8°22N, 

63°13W, USNM 80613–80614. Apuré: Hato La Guanota, 4 km W San Fernando de Apuré, 7°52N, 67°32W, 

TCWC 45153, 45167. Aragua: Cuyagua, 10°29N, 

67°42W, USNM 258124; El Limón, above Maracay, 

400 m, 10°18N, 67°38W, UMMZ 157138; Hacienda 

La Trinidad, near Maracay, 455 m, 10°17N, 67°37W, 

FMNH 35988–35989; Lake Valencia, northern shore, 

415 m, 10°11N, 67°45W, UMMZ 157102–157103; 

Maracay, 455 m, 10°15N, 67°36W, FMNH 190403, 

MCZ-A 50703; south of highway between Maracay and 

Valencia, 13 km W Maracay, 420 m, 10°16N, 67°38W, 

UMMZ 157104–157106; near Ocumare, 25 m, 10°28N, 

67°46W, UMMZ 122371(2), 171686–171689; road 

between Ocumare (inland village) and Rancho Grande, 

10°22N, 67°42W, UMMZ 157100; Parque Nacional 

Henri Pittier (Estación Biologica Rancho Grande), 910–

1,170 m, 10°22N, 67°41W, AMNH 70653, KU 132782, 

MZUSP 8083, UMMZ 122372, 157101; near Turmero, 

east of Maracay, 466 m, 10°14N, 67°29W (coordinates 

for Turmero), FMNH 27349. Bolívar: Ciudad Guayana, 

Parque Loefling, 100 m, 8°8N, 63°33W, USNM 229778; 

Santo Tomé de Guayana (= San Felix de Guayana), 50 m, 

8°2112N, 62°3910W, AMNH 81456–81457. Carabobo: El Central, 10 km NW, 25 m, 10°32N, 68°23W, 

USNM 216759; Río San Esteban, 10°2841N, 68°115W, 

UMMZ 55580. Distrito Federal: La Guaira, sea level, 

10°36N, 66°56W, USNM 22539, 27793. Falcón: near 

La Pastora, 12 km ENE Mirimiri, 220 m, 11°12N, 

68°38W, USNM 216729–216730; Municipio Mauroa, 

22 km SW Guajiro (= Goajiro) by car, AMU 3290; Pauji, 

Acosta district, 11°1N, 68°34W, MCZ-A 25985–25988; 

Riecito, Acosta district, MCZ-A 26143; Río Socopito, 

80 km NW Carora, 470 m, 10°28N, 70°44W, USNM 

216731; entre Municipios Sucre y Petit, Sector La Ceiba, 

Río Mitare, a la altura del pueblo de El Paso, 11°6N, 

69°55W, AMU 3234. Guárico: 8 km N, 2 km W Corozo 

Pando, ~8°30N, 67°35W, TCWC 45852; Sosa, 9°20N, 

67°12W, FMNH 125399, 176330. Mérida: Río Limones, 8°51N, 71°27W, ULABG 5090, 5111–5113, 5170, 

5304. Miranda: Araguita, 1 km S, 10°13N, 66°28W, KU 

166409–166410; Campo Grande de Caucagua, 10°17N, 

66°20W, MZUSP 58708–58711; Hacienda Bejuquero, 

1 km south Río Chico, 1 m, 10°19N, 65°58W, USNM 

216750–216753. Monagas: Caripito, Río Caripe, ~100 m, 

10°8N, 63°6W, AMNH 70650–70652; Escuela Granja 

Parcela, 8 km west-southwest Caripito, KU 117127. Portuguesa: Urbanización Andrés Eloy Blanco, Guanare, 

180 m, 9°3N, 69°45W, ELM 1167. Sucre: Río Cocollar, 

10°10N, 63°47W, FMNH 17769–17771; Guaraúnos, 

~100 m, 10°33N, 63°7W, KU 166404; Tocal, 11 km 

SSW Cumuna, 10°24N, 64°14W, KU 117122. Táchira:

La Fria Pueblo Nuevo, 8°13N, 72°15W, UMMZ 55582. 

Trujillo: near El Divide, 28–30 km NW Valera, 90 m, 

9°31N, 70°44W, USNM 216754–216758. Yaracuy: El 

Central, 10 km NW Urama, 25 m, 10°32N, 68°23W, 

USNM 216760; Palma Sola, 5 km SE, 10°36N, 68°34W, 

UMMZ 55581; 19 km NW of Urama, km 40, 25 m, 

10°37N, 68°24W, USNM 216727–216728, 216732–

216749. Zulia: Encontrados, 10 m, 9°30N, 72°14W, 

FMNH 2605; Setor Vera de Agua, km 12 on road El 

Vigía–Santa Barbara, 115 m, 8°422N, 71°437W, 

ULABG 5313.
We thank the following colleagues for assistance in examining 

specimens at their respective institutions (for institutional codes, 

see Sabaj Pérez, 2010) and/or responding to queries involving 

specimens at their institutions: Andrés Acosta-Galvis (Museo 

Universidad Javariana, Bogota), Ulisses Caramaschi (MNRJ), Ignacio De la 

Riva (MNCN), Marinus S. Hoogmoed (MPEG), Robert F. Inger (FMNH), 

John D. Lynch (ICNMHN), Charles W. Myers (AMNH), José P. Pombal, Jr. 

(MNRJ), Stephen P. Rogers (CM), Alan Resetar (FMNH), Gregory Schneider 

(UMMZ), Linda Trueb (KU), and Richard P. Vogt (Instituto Nacional de Pesquisas da Amazônia, Manaus).

The following colleagues generously provided help with difficult-to-pinpoint 

localities: Andrés Acosta-Galvis (Museo Universidad Javariana, Bogota), Erik 

Ahlander (NHRM), James P. Bogart (University of Guelph), José Enrique Castillo (IND), Maureen A. Donnelly (Florida International University), David 

Kizirian (AMNH), Enrique La Marca (Universidad de Los Andes, Mérida), 

John D. Lynch (ICNMHN), Charles W. Myers (AMNH), A. Stanley Rand (deceased, Smithsonian Tropical Research Institute), Alan Resetar (FMNH), José P. 

Rosado (MCZ), Jay M. Savage (Rana Dorada Enterprises, S.A.), John E. Simmons (KU), Neal Smith (Smithsonian Tropical Research Institute), and Paulo E. 

Vanzolini (MZUSP).

Materials or material queries were generously made available or responded 

to for this project by César Barrio-Amarós (Fundación AndígenA, Mérida), Ignacio De la Riva (MNCN), William E. Duellman (KU), Roberto Ibáñez (Smithsonian Tropical Research Institute), Travis La Duc (TNHC), Enrique La Marca 

(Universidad de Los Andes, Mérida), and John D. Lynch (Universidad Nacional 

de Colombia, Bogota).

Lee-Ann C. Hayek (National Museum of Natural History, Smithsonian Institution [NMNH-SI]) provided statistical advice. Addison Wynn (NMNH-SI) 

reworked Figure 9 for publication.

The research was supported by National Science Foundation award 

DEB-0342918.

Acknowledgments

Upper Shank, Outer Tarsus, 

and Sole of Foot Textures

After recording data on 1,002 specimens, it seemed that the amount of variation within population samples was about the same as that among samples 

for the surface textures of the dorsal surface of the shank, outer surface of the 

tarsus, and sole of foot. An analysis was undertaken to determine whether there 

was meaningful intersample variation in the characters involved.

Data were summarized for eight samples throughout the geographic distribution of the species complex: (1) 12 juveniles, 20 females, 4 males from Boca do 

Acre, Amazonas, Brazil; (2) 28 juveniles, 7 females, 1 male from Ilha do Maracá, 

Roraima, Brazil; (3) 10 juveniles, 4 females, 5 males from Hacienda El Ceibal, 

Bolívar, Colombia; (4) 30 juveniles, 3 females, 2 males from Beltrán, Cundinamarca, Colombia; (5) 23 juveniles, 4 females, 5 males from between Santa Marta 

and Mamatoco, Magdalena, Colombia; (6) 40 juveniles, 5 females, 3 males from 

a few localities along the Río Chucunaque, Darién, Panama; (7) 46 juveniles, 1 

female, 1 male from Pakitza and Tambopata, Madre de Dios, Peru; (8) 10 juveniles, 5 females, 3 males from 19 km northwest Urama, Falcón, Venezuela. Data 

were summarized for juveniles, females, and males for the following states of a 

surface shagreen: none, weak shagreen, and shagreen. These states were scored 

separately for the shank, tarsus, and foot. The shagreen texture is sandpaper-like, 

discernable only under magnification, with the skin surface just moist so the light 

reflects off the small protuberances comprising the shagreen. Shagreened surfaces 

seem to be sloughed off in specimens that are not well preserved. Data were also 

summarized for juveniles, females, and males for the following tubercle states: 

none, few, scattered, and many. These states were summarized separately for the 

shank, tarsus, and foot. The tubercles involved in well-preserved specimens are 

small, black keratinized protuberances. In less than well-preserved specimens, the 

outer keratinized covering may be lost leaving (smaller) white tubercles. These 

structures are consistently interpretable only under magnification.

The character states form a continuum (e.g., for shagreen, three states were 

coded as none, weak shagreen, and shagreen). If a given specimen was scored as 

Appendix 1: Observer Error Assessment 

for Morphological Data
4 8 • smithsonian contributions to zoology

“none” in the first data-taking session and “none” in the 

second data-taking session, there is no difference in the 

scoring between the two sessions for shagreen for the specimen involved. If a given specimen was scored as “none” 

in the first data-taking session and “weak shagreen” in the 

second data-taking session, that is considered to be a onestep difference; if the specimen was scored as “none” in 

the first data-taking session and “shagreen” in the second 

session, that is considered to be a two-step difference. The 

two-step difference represents a larger observer error than 

a one-step difference. The term step does not have any cladistic connotation as used in the following analyses.

Examination of the distribution of occurrences of 

character states among the eight samples indicates that 

there are no occurrence differences among juveniles, females, and males within each locality. The distributions 

of states are so obvious that statistical analyses are not 

required (data available from WRH upon request). The 

data appeared as though there might be meaningful variation among the localities for both the shagreen and tubercle characters for the three different surfaces involved. 

As the character states are defined qualitatively, subjective 

interpretation could well vary from data-taking session to 

data-taking session by the same observer. As there were 

large time intervals between bouts of data taking for this 

project, the interpretations could well be quite different. 

To evaluate the impact of observer error, USNM specimens 

for two of the samples that were involved in the first bout 

of data taking (Amazonas, Brazil, and Cundinamarca, Colombia) were rescored for the characters involved. Thus 

each specimen had two character states recorded. For example, the upper shank surface on the first observation 

might have been scored as “none” and when the second 

observation was made on the same specimen, it could have 

scored as “shagreened.” Alternatively, the first and second 

evaluations of the same specimen could have been “none” 

and “none.” The results are reported separately for the 

two characters.

Shagreen

For the upper shank surface, 33% of the individuals 

were scored as having the same state, 10% were scored as 

having a 1-step difference (all of the differences involved 

were those between the Weak Shagreen and Shagreen 

states), and 57% of the individuals were scored as having 

a 2-step difference (all involved No Shagreen and Shagreen 

states). For the outer tarsal surface, 15% were scored as 

having the same state, 33% with a 1-step difference (all involved No Shagreen and Weak Shagreen states), and 52% 

a 2-step difference (all involved No Shagreen and Shagreen 

states). For the sole of the foot surface, 92% were scored 

as having the same state (No Shagreen) and 8% as having 

a 1-step difference (all involved No Shagreen and Weak 

Shagreen states). These results indicate that observer error 

masks any differences that occur among individuals for 

the states involved. The only conclusion that can be supported with the large observer error is that the sole of the 

foot generally lacks a shagreen in members of this species 

complex and that most individuals have shagreened dorsal 

shank and outer tarsal surfaces.

Tubercles

For the upper shank surface, 49% of the individuals 

were scored as having the same state, 47% were scored 

as having a one-step difference (all involving the scattered 

tubercles and many tubercles states), and 4% as having a 

two-step difference (all involving the no tubercles and scattered tubercles states). For the outer tarsal surface, 54% 

of the individuals were scored as having the same state, 

39% with a one-step difference (mostly involving the few 

tubercles and scattered tubercles states), and 6% of the 

individuals with two-step differences (all involving the no 

tubercles and scattered tubercles states). For the sole of the 

foot surface, 37% of the individuals were scored as having 

the same state, 60% were scored as having a one-step difference (mostly involving the no tubercles and few tubercles states), and 2% with a two-step difference (involving 

the no tubercles and scattered tubercles states). Observer 

error for the tubercle states has less of an impact than for 

the shagreen states but is still considerable. Ninety-four to 

98% of all error falls within a one-step difference of the 

four-state continuum of categories. All of the differences 

observed in distribution of states among the eight geographic samples occurred between adjacent states, however, indicating that the variation is not meaningful when 

observer error is accounted for.

Owing to observer error and limited variation observed in the eight geographic samples, it is clear that geographic variation cannot be demonstrated convincingly 

for the texture characters involved. Consequently, data for 

these characters were not taken after February 2005.

The following characterization can be made for the 

Leptodactylus bolivianus complex. Shagreened surfaces 

are most commonly observed on the entire dorsal shank 

and outer tarsal surfaces. The sole of the foot usually does 

not have any shagreen; however, in the few cases where 

shagreen is present, it is limited to the proximal half of the 

fifth metatarsal region of the sole of the foot. Almost all 
number 635 • 4 9

individuals have small black keratinized (or white) tubercles rather evenly distributed on the dorsal surface of the 

shank and outer surface of the tarsus. The sole of the foot 

may lack tubercles entirely, although most specimens have 

them. When black (or white) tubercles are present, they 

are typically smaller than those found on the dorsal shank 

and outer tarsal surfaces and characteristically occur only 

on the outer half of the sole of the foot.

Methodology for Remaining

Characters Exclusive of Male

Secondary Sexual Characters

The same series of 36 specimens from Beltrán, Cundinamarca, Colombia evaluated for observer error in texture characters (preceding section) were used to evaluate 

observer error for sex, belly pattern, dorsal pattern, lip 

stripe pattern, posterior thigh pattern, upper shank pattern, longitudinal body folds, and male secondary sexual 

characteristics. Data were originally recorded for these 

specimens at the beginning of the study prior to 27 August 

2004. Character states were defined as written statements 

(e.g., for upper shank textures) or, where written statements were not as clear as sketches, drawings were rendered. As data taking proceeded, if a character was similar 

but not identical to a sketch, a brief statement characterizing the difference was added (Figure A1.1). Some pattern states were added after 27 August 2004. In theory, as 

any new pattern was encountered during the study, a new 

standard was prepared for it. Thus any specimen that was 

examined early in the study should not be scored as having a standard subsequently added if the specimen were 

to be reexamined. In order to test this aspect along with 

observer error in general, the Beltran series was rescored 

on 29–30 August 2005. In order to determine how repeatable the character states were scored over a short period of 

time, the Beltrán series was scored a third time on 13–14 

September 2005.

Sex

The scoring for sex was anticipated to be invariant 

among the observation recording sessions with the exception of a few of the larger juveniles that were not examined 

internally the first session and scored simply as juveniles 

but were examined internally the second session and were 

recorded as either juvenile females or juvenile males.

Six specimens were scored differently between the first 

and second and first and third data-recording sessions on 

the basis of dissections made during the second session. 

These differences are not considered to be observer errors.

Specimen USNM 145715 (76 mm SVL) was scored as 

an adult female in sessions 1 and 2 but a juvenile female 

in session 3. Reexamination of the specimen indicates that 

with the dissection view available during sessions 1, 2, and 

3, only the ovary was visible. Further dissection was required to expose an oviduct, which is curly; thus the specimen is an adult female following the criteria used in this 

study.

Specimen USNM 145716 (71 mm SVL) was scored as 

a juvenile female in sessions 1 and 3 and an adult female 

in session 2. Reexamination of the specimen indicates that 

is has a straight oviduct and the ovaries are expanded containing small white ova. The straight oviduct condition is 

used to define the juvenile female condition in this study.

Specimen USNM 145721 (63 mm SVL) was scored as 

juvenile in session 1, a juvenile female in session 2, and a 

juvenile male in session 3. A fourth examination to determine which of the results in sessions 1, 2, or 3 was correct 

clearly indicated that the specimen has a straight oviduct 

with expanded ovaries with small white ova upon reexamination. The specimen is unquestionably a juvenile female; 

there had to be a transcription recording error in session 

3, where the condition observed was not the condition recorded during session 3.

Specimen USNM 145724 (59 mm SVL) was recorded 

as a juvenile in session 1, a juvenile female in session 2, 

and an adult female in session 3. Reexamination of the 

specimen indicates that the oviduct is curly and the ovaries are expanded with moderate sized white ova, an adult 

female according to the criteria in this study.

In conclusion, there are two kinds of observer error 

detected in the reexamination process for determination 

of sex. The first is making different conclusions about 

whether near-adult sized females are adults or juveniles 

(5% scoring error). The second kind of error is a data 

transcription error (1% error).

Belly Pattern

A total of 15 belly pattern standards were recorded for 

the three Beltrán sample data-taking sessions. Two pattern 

standards, added after the initial session, were scored with 

a 24% frequency in sessions 2 and 3. This indicates that 

not all variation thought to be important was recognized 

at the beginning of the study.

Only 14% of the standards were scored as the same 

between sessions 1 and 2, and 17% between session 1 and 

3; whereas 39% of the standards were identically scored 
5 0 • smithsonian contributions to zoology

FIGURE A1.1. Example of design pattern. First page of belly pattern standards used in study to score character states. The original sheet had 

written annotations defining additional standards in relation to the pattern involved: A-1, dark and light areas about equal; A-2, mostly dark 

with few light spots, more light spots posteriorly; A-3 as A-1, except darker to lighter gradient anteriorly-posteriorly; B-1, mottling on posterior, 

almost uniform dark anteriorly; B-2, uniformly mottled as on posterior; B-3, gradient of anterior pattern posteriorly; B-4, posterior pattern with 

anterior-posterior gradient; B-5, anterior pattern on entire belly; C-1, anterior mottling continuous across middle; C-2, almost lacking pattern; 

C-3, uniformly darkly mottled (fine); C-4, uniformly lightly mottled (fine); D-1, mostly light; D-2, indistinct anteriorly; E-1 (only anterior portion filled in), fewer spots; E-2, very few spots; F, virtually no pattern; F-1, just a few small blotches lateral-most; F-2, uniformly dark; G-1, very 

little in middle; G-2, groups of melanophores instead of discrete marks; G-3, pattern anteriorly only; H-1, finer pattern; H-2, almost uniform 

dark anteriorly; H-3, anterior-posterior gradient, lighter posteriorly; I-1, encroachment only a bit laterally behind arms; I-2, pattern extends to 

posterior belly; J-1, + a few [blotches] on lateral belly; J-2, everywhere except center of belly; J-3, light speckling over entire belly.
number 635 • 5 1

between sessions 2 and 3. Thus there is more consistency 

in scoring over a shorter time period (sessions 2 and 3) 

than for longer time periods between data taking (sessions 

1–2, 1–3).

Given the low repeatability of scoring the 15 standards 

recorded for the Beltrán sample, the following two-step 

procedure was used to combine the standards to maximize 

repeatability on scoring of the data.

The first step was to visually inspect the 15 standards 

and combine states that were most similar. This resulted in 

recognition of seven states: (1) anterior mottling only, (2) 

belly mottled with an anterior–posterior gradient, (3) uniform mottling over the entire belly, (4) belly with anterior 

speckling only, (5) belly speckled with an anterior–posterior gradient, (6) belly uniformly speckled, and (7) no 

pattern (no melanophores on belly). The Beltrán sample 

data were rescored using these seven states. This resulted 

in 17% exact matches between session 1 and session 2, 

25% between session 1 and session 3, and 56% between 

session 2 and session 3.

The second step was to examine the raw occurrence 

data from step 1 to determine which further combinations 

of character states would result in the greatest repeatability in the data set. Combining mottling and speckling 

states did not appreciably improve repeatability. However, 

combining these states with either of two other combinations did result in noticeable improvement. When mottling 

+ speckling were combined with anterior only + anterior–

posterior gradient states, the states were identical in 50% 

of the time between sessions 1 and 2 and between sessions 

1 and 3, and 80% between session 2 and 3 scorings. When 

mottling + speckling were combined with anterior–posterior gradient + uniform states, the states were identical 

42% of the time between session 1 and 2 data, 53 % between session 1 and 3 data, and 69% between session 2 

and 3 data.

Thus, for the Beltrán data, the following final states 

are recognized: (1) belly speckled or mottled anteriorly 

only or with an anterior–posterior gradient, (2) belly 

speckled or mottled with the same intensity over the entire 

belly, and (3) belly patternless (no melanophores).

Finally, the total data set was evaluated visually using 

the above results as a guideline. Thirty-four belly pattern 

states occur in the total data set. Among the states that 

were not recorded in the Beltrán samples are two distinct 

features: spotted bellies and boldly mottled bellies. However, there are very few times the spotted belly state was 

recorded. The variation found, including these new features, result in the recognition of the following states used 

in analyses of belly patterns: (1) belly speckled, spotted, or 

mottled anteriorly only or with an anterior–posterior gradient, (2) belly boldly mottled anteriorly only or with an 

anterior–posterior gradient over the entire belly, (3) belly 

speckled or mottled with the same intensity over the entire belly, (4) belly boldly mottled with the same intensity 

over the entire belly, and (5) belly without a pattern (no 

melanophores). These states are assumed to have around 

50% repeatability over the entire study and around 75% 

repeatability within data-taking sessions at single institutions or for multiple institutions visited consecutively during the same trip. Thus any belly pattern differences in 

data comparisons must exceed 50% (for adequate sample 

sizes) to be meaningful.

Dorsal Pattern

Eight dorsal pattern standards were recorded during 

the three Beltrán sample data-taking sessions. The same 

standards were used for all sessions. One transcription 

error was made where a nonexistent dorsal pattern standard was recorded.

Fifty-six percent of the standards were recorded as 

identical between sessions 1 and 2, 58% between sessions 1 and 3, and 75% between sessions 2 and 3. There 

is greater consistency in scoring over a shorter time period 

(sessions 2–3) than over a longer time period (sessions 

1–2, 1–3).

The dorsal pattern standards were examined visually, and the most similar patterns were combined to see 

if the increase in repeatability offset the loss of potential 

information on variation. The following three states were 

recognized and used to recode the three session data: (1) 

dorsum uniform between interocular bar (when present) 

and sacrum; (2) single chevron between interocular bar 

and sacral chevron (when present) without additional 

spots lateral to dorsal chevrons; and (3) complex pattern 

with two dorsal chevrons and spotting all over dorsum, 

chevrons often elongate and fused with each other and interorbital bar. The recoded data yield 77% exact match of 

states between data for sessions 1 and 2, 67% between session 1 and 3 data, and 82% between session 2 and 3 data.

The increase in repeatability of the combined standards is considered worth the cost of losing information 

on variability of the dorsal patterns. However, further 

reduction of the states would be too great a loss of information on variability. For dorsal pattern variation, any 

adequate size sample comparisons have to differ by more 

than 33% in order to be meaningful.

The three states defined above encompass the variation recorded in the total data set.
5 2 • smithsonian contributions to zoology

Lip Stripe

Two features were evaluated for the lip stripe: the 

overall lip stripe pattern and whether a distinct light patch 

was present immediately below the eye.

Eighteen pattern states were recorded for the overall 

lip stripe pattern in the three Beltrán sample data sets. 

Nine percent of the first and second session data elements 

were scored identically, 29% were identical between session 1 and 3 data, and 42% were identical between session 2 and 3 data. One standard was added after the first 

session, which was scored at a 3% occurrence in the combined session 2 and 3 data.

The lip pattern standards were examined visually and 

combined to minimize mismatches in the original pattern 

scoring results. Four states were used to recode the data 

for the three sessions: (1) a noticeable broad light stripe 

extending from the tip of the snout to through the commissural gland, bordered below by continuous or broken 

darker markings on the upper lip and either bordered 

above by a dark line under the eye or not; (2) a narrow 

light stripe from at least the nostril to the lower posterior 

end of the eye; (3) a narrow light stripe from below the 

front of the eye through the commissural gland; and (4) 

central lip region uniform (about same color intensity as 

background dorsal color) with or without a dark stripe 

under the eye and/or dark bars/mottling on the upper lip. 

Identical pattern states were scored 53% of the time for 

the session 1 and 2 data, 69% for session 1 and session 3 

data, and 57% for session 2 and session 3 data.

Further reduction of states would result in very little 

variation in the data set. For the overall lip pattern variation, any adequate sample size comparisons have to differ 

by more than 50% in order to be meaningful.

Three conditions of expression of a light patch under 

the eye (subocular light spot) were recorded in the three 

Beltrán data sets: (1) distinct, (2) indistinct, and (3) absent. Ninety-four percent of the first and second session 

data states were scored identically, 94% were identical between sessions 1 and 3 data, and 92% were identical between sessions 2 and 3 data. The level of observer error is 

considered acceptable. There are no further states for this 

character in the entire data set. For light subocular patch 

variation, adequate sample size comparisons have to differ 

by more than 10% to be meaningful.

Posterior Thigh Pattern

Thirteen posterior thigh pattern standards were scored 

for the three Beltrán data-recording sessions. One pattern 

was added after the first session but was not recorded in 

either the session 2 or 3 data. Two transcription errors 

were made where a nonexistent standard was recorded.

Identical pattern standards were recorded 28% of the 

time between sessions 1 and 2, 31% between sessions 2 

and 3, and 31% between sessions 2 and 3. The scoring 

was quite consistent, but with high observer error in scoring, among sessions.

The standards were examined visually to combine the 

most similar patterns and to minimize scoring differences 

found in the raw scoring results. The following five states 

were recognized and used to recode the three session data: 

(1) posterior thigh with a uniform or variable dispersion 

of melanophores (weakly to moderately mottled) over the 

entire surface of the posterior thigh; (2) posterior thigh 

with a darker patch with small, irregular light spots on 

the distal half of the thigh; (3) posterior thigh with a dark 

vermiculated pattern on the distal half of the thigh; (4) 

posterior thigh pattern ranges from boldly mottled with 

large light irregular spots or vermiculations on a dark 

background to the light markings being more extensive 

than the dark areas, the light markings sometimes coalesced; and (5) posterior thigh uniformly dark with only 

a very few light irregular markings. Identical states of the 

rescored data between session 1 and session 2 occurred 

69% of the time, 92% of the time between sessions 1 and 

3, and 64% of the time between sessions 2 and 3.

Patterns 2 and 3 defined above were only scored twice 

each in the entire data set. Pattern 5 defined above (created 

from examination of specimens of the L. latrans complex) 

was scored once in the rescoring of the Beltrán sample 

specimens. In addition, two other patterns were scored 

once in the entire data set. All of these single- or doubleoccurrence states are combined with their closest patterns 

with the result that only two states are recognized for this 

character: (1) posterior thigh with a uniform or variable 

dispersion of melanophores (weakly to moderately mottled) usually over entire surface of posterior thigh, and (2) 

posterior thigh boldly mottled with large light irregular 

spots/vermiculations on a dark background to the light 

marks being more extensive than the dark areas, the light 

markings sometimes coalesced. 

Adequate sample size comparisons have to differ by 

more than 33% in order to be meaningful.

Dorsal Shank Pattern

Six patterns were scored in the three data-recording 

sessions of the Beltrán sample. No patterns were added 

after the first session.

Identical pattern standard states were recorded 42% 

of the time in sessions 1 and 2 data, 25% in sessions 1 and 
number 635 • 5 3

3 data, and 69% in sessions 2 and 3 data. Scoring between 

sessions was quite inconsistent.

Only minimal combining of pattern states was possible by visual examination of the standards, resulting in 

recognition of the following three character states: (1) uniform, (2) short dark transverse bars extending less than 

halfway across shank, and (3) at least one dark transverse 

bar extending halfway or more across the shank. Identical 

states in the rescored data occurred 50% of the time in 

sessions 1 and 2 data, 31% in sessions 1 and 3 data, and 

72% in sessions 2 and 3 data. These values are only marginally better with respect to the values based on uncombined pattern character states. Further combining of states 

would result in little variation of the upper shank patterns. 

There is no additional variation for this character in the 

entire data set. Adequate sample size comparisons have to 

differ by more than 70% in order to be meaningful.

Dorsolateral Folds

Six states were recognized in the Beltrán specimen 

data. One state was added after the first data-recording 

session, which was recorded in 11% of the session 2 and 

3% of the session 3 data.

Fifty percent of the specimens were recorded with 

identical states between sessions 1 and 2, 47% between 

sessions 1 and 3, and 53% between sessions 2 and 3.

The initial states were combined into two states to determine whether the increase in repeatability was worth the 

loss of information concerning variation: (1) dorsolateral 

fold distinct and (a) continuous from supratympanic fold 

to end of body, or (b) with a brief interruption of the fold, 

or (c) the fold is distinct only from the supratympanic fold 

to the sacral region; and (2) dorsolateral fold with several 

interruptions to indiscernible (including lack of fold probably due to poor preservation). The rescored comparisons 

resulted in 78% exact matches between data sessions 1 

and 2, 65% between sessions 1 and 3, and 76% between 

sessions 2 and 3. The gain in repeatability is considered 

worth the loss of information concerning variability.

The redefined states encompass all the variation observed in the entire data set. Differences in dorsolateral 

fold states must exceed 33% in adequate sample sizes to 

be meaningful.

Lateral Folds

Six states were recognized in the three data-taking sessions. One state was added after the first session and was 

recorded in 3% of the individuals for both the session 2 

and 3 data.

Identical states were scored 39% of the time between 

session 1 and session 2 data, 44% between sessions 1 and 

3, and 53% between sessions 2 and 3.

The initial states were combined into two states: (1) 

lateral fold distinct or slightly interrupted from supratympanic fold to leg and (2) lateral fold distinct only in groin 

region, indistinct overall, or indiscernible (probably due 

to poor preservation). The rescored data between sessions 

1 and 2 were identical in 61% of the comparisons, 71% 

between sessions 1 and 3, and 88% between sessions 2 

and 3. The increase in repeatability is considered worth 

the loss of variation information.

There is no further variation in this character in the 

entire data set. Differences in lateral fold states must exceed 40% among adequate size samples to be meaningful.

The new states are used in subsequent analyses. 

There are two extremes of repeatability of character 

states represented by belly and lip patterns at one extreme 

and the light subocular patch at the other. The belly and 

lip patterns involve several features that together represent a total gestalt. In retrospect, WRH focused on different components of the belly and lip pattern standards 

at different times. For example, during one session more 

attention might have been focused on the nature of the 

markings on the upper lip adjacent to the jaw; during another session the focus might have been on how distinctive the broad light stripe was between the upper lip and 

the eye; and yet another session may have focused more 

on the nature of the dark markings immediately below 

the eye. Such unintentional shifts in focus could explain 

why there was more consistency in observations made 

over short periods of time and much greater inconsistency 

when data were taken after a long hiatus between sessions. In contrast, the light subocular patch is a single 

feature for which the variation was recorded much more 

consistently.

There are at least two ways that complex patterns 

could be analyzed differently than was done in this study 

to minimize observer error. The first would be to take all 

the data in one continuous session, where on a day-to-day 

basis, there would likely be carryover of which features 

in complex patterns were being focused on. The second 

would be to pick a series of specimens that likely encompass the range of variation in the entire study sample and 

prepare a series of sketches that represent that range of 

variation for each general character to be evaluated, such 

as belly pattern. On the basis of those sketches, individual 

features could be identified and character states defined 

that would then be used to evaluate the entire data set. 

The second approach would seem more effective to minimize observer error.
5 4 • smithsonian contributions to zoology

Male Secondary 

Sexual Characters

There are only two adult males in the Beltrán sample. 

The only variation among the three data sessions was a 

one-step difference out of a four-state range in evaluation 

of the degree of male arm hypertrophy.

The male secondary sexual characteristics can be 

defined and described in terms of individual features as 

follows.

Male Arm. (1) Upper arm not hypertrophied 

and (2) upper arm hypertrophied.

Male Lower Jaw. (1) Male lower jaw lacking keratinized tubercles and (2) male lower jaw with keratinized tubercles.

Male Chest. (1) Male chest lacking a patch 

of keratinized tubercles, (2) male chest with a central 

patch of keratinized tubercles, and (3) male chest with a 

central patch and two elongate lateral patches of keratinized tubercles just posterior to the central patch.

Male Thumb. (1) Male thumb with one 

weakly chisel-shaped keratinized spine, (2) male thumb 

with one well-developed chisel-shaped keratinized spine, 

and (3) male thumb with two round keratinized spines. 

The spines have keratinized sheaths at least during the 

reproductive season. Spines lacking sheaths still demonstrate the differences between the chisel-shaped and 

rounded features.

The above definitions would be expected to have the 

same repeatability as those for the light subocular patch 

character. Erring on the conservative side, adequate sample size comparisons for the male secondary sexual characters should differ by more than 15% to be meaningful.
LEPTODACTYLUS BOLIVIANUS COMPLEX

LOCALITY DATA USED IN MOLECULAR ANALYSIS

Leptodactylus bolivianus. USNM 268966 (USFS 152368)—PERU: Madre 

de Dios, Tambopata Reserve, 12°50S, 69°17W, 66 m (collection of Jiri 

Moravec); BOLIVIA: Pando, settlement of Nacebe, 11°00S, 67°25W.

Leptodactylus guianensis. AMNH145130–131 (JC 7056–57)—GUYANA: 

Corontyne Region, Dubulay Ranch on the Berbice River, 5°405N, 57°5132W, 

UC 128, 130; BRAZIL: Roraima, proximity of Vila Surumu, 04°1140.9N, 

60°4725.1W.

Leptodactylus insularum. MUJ 2187—COLOMBIA: no other data. JDL 

24816—COLOMBIA: Sucre, San Marcos, vereda La Florida, 8°37N, 75°11W, 

45 m. JDL 24887—COLOMBIA: San Marcos, finca Crocodilia, 8°36N, 

75°09W, 34 m. JDL 26573, JDL 26591—COLOMBIA: Cordoba; Lorica, estacion pisicola CVS, 9°13N, 75°50W, 20 m. CH 4955–56—PANAMA: Panamá, 

Río Indio, camino hacia Las Minas. ULABG 5111–13, 5304—VENEZUELA: 

Mérida; Río Limones.

Appendix 2. Molecular Sample Locality Data

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Boulenger, G. A. 1898. A List of the Reptiles and Batrachians Collected by the Late Prof. 

L. Balzan in Bolivia. Annali del Museo Civico di Storia Naturale di Genova, Series 

2, 19:128–133.

Capocaccia, L. 1957. Catalogo dei Tipi di Anfibi del Museo Civico di Storia Naturale di 

Genova. Annali del Museo Civico di Storia Naturale di Genova, 69:208–222.

Charif, R. A., C. W. Clark, and K. M. Fristrup. 2004. Raven 1.2 User’s Manual. Ithaca, 

N. Y.: Cornell Laboratory of Ornithology.

Duellman, W. E. 1997. Amphibians of La Escalera Region, Southeastern Venezuela: Taxonomy, Ecology, and Biogeography. Scientific Papers, Natural History Museum, The 

University of Kansas, 2:1–52.

———. 2005. Cusco Amazónico. The Lives of Amphibians and Reptiles in an Amazonian Rainforest. Ithaca, N.Y.: Cornell University Press.

Engelman, L., S. N. Badashah, and R. V. Nath. 2004. “Discriminant analysis.” In Statistics, ed. Anonymous, pp. I301–I358. Richmond, Va.: SYSTAT Software, Inc.

Fabrezi, M., and P. Alberch. 1996. The Carpal Elements of Anurans. Herpetologica,

52:188–204.

Felsenstein, J. 1985. Confidence Limits on Phylogenies: An Approach Using the Bootstrap. Evolution, 39:783–791. doi:10.2307/2408678.

Fouquette, M. J., Jr. 1960. Call Structure in Frogs of the Family Leptodactylidae. The 

Texas Journal of Science, 12:201–215.

Gosner, K. L. 1960. A Simplified Table for Staging Anuran Embryos and Larvae with 

Notes on Identification. Herpetologica, 16:183–190.

Hayek, L.-A. C., W. R. Heyer, and C. Gascon. 2001. Frog Morphometrics: A Cautionary 

Tale. Alytes, 18:153–177.

Heyer, W. R. 1969. Studies on Frogs of the Genus Leptodactylus (Amphibia, Leptodactylidae). V. Taxonomic Notes on L. latinasus, rhodonotus, romani, and wuchereri. 

Herpetologica, 25:1–8.

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