New Species of Physalaemus Fitzinger, 1826 from Southern Bahia, Brazil (Anura, Leptodactylidae)

Author(s) :Carlos Alberto Gonalves Cruz and Bruno V. S. Pimenta Source: Journal of Herpetology, 38(4):480-486. 2004. Published By: The Society for the Study of Amphibians and Reptiles DOI: URL: http://www.bioone.org/doi/full/10.1670/214-02A

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Journal of Herpetology, Vol. 38, No. 4, pp. 473480, 2004 Copyright 2004 Society for the Study of Amphibians and Reptiles

Reproduction and Sexual Size Dimorphism in the Lacertid Lizard Pedioplanis burchelli (Sauria: Lacertidae) in South Africa
W. T. NKOSI,1 N. J. L. HEIDEMAN,1,2
1

AND J.

H. VAN WYK3

Department of Zoology and Entomology, University of the Free State (Qwaqwa Campus), Private Bag X13, Phuthadithjaba 9866, South Africa 3 Department of Zoology, University of Stellenbosch, Private Bag Matieland, Stellenbosch, South Africa

ABSTRACT.Pedioplanis burchelli is a small oviparous lacertid lizard that inhabits rocky montane areas in the highveld grassland biome of South Africa. We studied its reproductive cycle by monthly monitoring of gravimetric, morphometric, and histological changes of the reproductive system of males and females throughout 1999. Reproduction took place in spring/summer, and gonadal activity of males and females was well synchronized and prenuptial. Photoperiod was the best predictor of seasonal uctuations in testes mass and rainfall in the case of ovarian seasonality. Neither vitellogenic follicle count nor oviducal egg count were correlated with snoutvent length. Sexual size dimorphism with respect to certain body parts is explained either in terms of sexual selection, differential growth, or intraspecic food niche segregation. The absence of a signicant difference in incidence in damaged tails between males and females may be indicative of similar levels of predation pressure.

Lizard species diversity in Southern Africa ranks among the highest in the world, with an endemicity of more than 92% (Bauer, 1999). However, life history is larger unstudied in most species, even entire families (for example, the Lacertidae). The Lacertidae comprises nine genera of which Pedioplanis comprises 11 species, nine of which are endemic to South Africa (Branch, 1998). Apart from a detailed study of two Namib Desert lacertid species belonging to the genus Meroles (Louw and Holm, 1972; Goldberg and Robinson, 1979), existing knowledge of southern African lacertid reproductive biology is anecdotal and based mostly on brief comments in taxonomic works such as De Waal (1978) and Branch (1998). This study is the rst detailed account of reproduction in a South African lacertid lizard. Pedioplanis burchelli is insectivorous and cryptically colored and inhabits rocky montane areas in highveld grassland biome where individuals take shelter mainly under at sandstone rock slabs (Branch, 1998). The aims of the present study were to describe the reproductive cycle of males and females and how they correlate with each other and with certain environmental variables. In addition, sexual size dimorphism was investigated and the incidence of damaged tails used as possible indicator of predation pressure experienced by males and females.

2 Corresponding Author. E-mail: heidemannj.sci@ mail.uovs.ac.za

MATERIALS AND METHODS Study Area and Climate.The study area was situated in the highveld grassland biome of South Africa, in the Phuthadithjaba area of the northeastern Free State Province between 288299S latitude and 288499E longitude, at an altitude of 1850 m above sea level. Rain falls mainly in summer and varies from 250 to 500 mm per year (Rutherford and Westfall, 1986). Weather data for 1998 and 1999 (Fig. 1) were obtained from Bethlehem (about 75 km northwest of the study area), the nearest weather station. Collecting and Laboratory Procedure.A minimum of six adult males (only three in April) and females were collected around the middle of each month from January through December 1999. Lizards were killed by freezing within 48 h of capture, mass recorded to the nearest 0.1 g, and snoutvent (SVL) measured to the nearest 0.1 mm. In males, testes and ductuli epididymides were excised, mass recorded to the nearest 0.01 g, xed and stored in Bouins solution. Testis length was measured to the nearest 0.1 mm. In females, ovaries and oviducts were removed and mass recorded to the nearest 0.01 g. The diameter of the four largest ovarian follicles was measured to the nearest 0.1 mm, and their developmental stages described following the criteria of Van Wyk (1984). Appearance of vitellogenic follicles, corpora lutea, and oviducal eggs was recorded. Oviducal eggs were counted to estimate clutch size. The reproductive cycle was studied by observing gravimetric, morphometric, and histological changes (males only) of gonads and accessory ducts throughout the year for each sex. Histo-

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W. T. NKOSI ET AL. lutea present (November) but not in (January); (5) Late postovulatory (II), Early vitellogenic follicles (diameter: from 1.82.0 mm); oviducal eggs or corpora lutea present (November); (6) Preovulatory period (II), Late vitellogenic follicles (diameter: from 3.06.2 mm); no oviducal eggs or corpora lutea (NovemberDecember); (7) Postovulatory (III), Hydrated ovarian follicles (diameter: 2.0 mm); no oviducal eggs but corpora lutea present (December). The following body dimensions were also compared between the sexes: SVL, TL (using specimens showing no signs of tail damage or regeneration), head length (HDL), width (HDW), and height (HDH), as well as forelimb (FL) and hindlimb length (HL). All measurements were taken with a digital sliding vernier caliper to the nearest 0.1 mm. HDL was measured from the posterior edge of the ear opening to the tip of the snout, whereas HDW and HDH were measured at an imaginary plane running vertically through the center of the ear openings. FL and HL comprised the respective combined lengths of the upper and lower limb elements, hand/foot and longest digit. The female:male ratio (FMR) of Fitch (1981) was used to calculate the degree of dimorphism. The entire sample was examined for damaged tails and signs of regeneration for comparison between males and females. Specimens with freshly damaged tails were not considered to ensure that ones with capture injuries were not included in the analyses. Data Analysis.All means are given as 6 1 SE. Data were tested for normality using the Kolmogorov-Smirnov test and for homogeneity of variances using Bartletts test (Sokal and Rohlf, 1981) prior to analyses. Data on epididymus mass, epithelium height and diameter as well as ovarian mass, oviducal mass and follicle diameter did not meet assumptions of normality. Kruskal-Wallis nonparametric analysis of variance was therefore used to test for signicant seasonal variation in those parameters, followed by Dunns nonparametric multiple comparisons test for ranked data (Heath, 1995). The number of groups compared (k) using the latter test was eight, giving a test statistic, Q, of 3.124 at P 0.05. Covariation between reproductive parameters and environmental factors was analyzed using Spearmans rank correlation analyses and stepwise multiple regression analyses with monthly testis mass and ovarian mass as dependent variables and the climatic factors (maximum and minimum ambient temperature, photoperiod, and rainfall) as independent variables. In the case of the morphometric comparisons, all variables were log-transformed before regression analysis, and ANCOVA were carried out. Least-squares regression analysis of logtransformed variables against log total body

FIG. 1. Mean maximum and minimum ambient temperature, photoperiod, and rainfall for 1998 and 1999 recorded at Bethlehem.

logical sections were made using the method described by Prophet et al. (1994), which involved staining sections with Harriss haematoxylin and eosin. Sections were taken at 8lm and the diameter of ve randomly selected crosssections of the seminiferous tubules measured per testis to the nearest 1 lm using a microscope graticule. In the case of the epididymus, changes in the height of its epithelium were measured and the presence of spermatozoa in its lumen recorded. Staging of spermatogenic activity followed the classication system of Licht (1967), which comprises the following stages: Stage 1, seminiferous tubules involuted with only spermatogonia; epididymus atrophic and empty; Stage 2, seminiferous tubules with primary spermatocytes; epididymus atrophic and empty; Stage 3, secondary spermatocytes and early spermatids present; epididymus atrophic and empty; Stage 4, transforming spermatids with a few spermatozoa; epididymus atrophic and empty (but others with few spermatozoa); Stage 5, spermatozoa abundant with maximum spermiogenesis; epididymus hypertrophied and many spermatozoa; Stage 6, spermatozoa abundant but spermatocytes and spermatids greatly reduced; epididymus atrophic and empty; Stage 7, seminiferous tubules involuted with only spermatogonia; epididymus hypertrophied and empty. Ovarian activity was staged according to the classication of Van Wyk (1984) which is as follows: (1) Nonreproductive stage, Only hydrated ovarian follicles (diameter: 1.8 mm); no oviducal eggs or corpora lutea (MarchAugust); (2) Early vitellogenic stage, Early vitellogenic follicles (diameter: from 1.82.0 mm); no oviducal eggs or corpora lutea (SeptemberOctober); (3) Preovulatory stage (I), Late vitellogenic follicles (diameter: from 2.12.9 mm); no oviducal eggs or corpora lutea (October); (4) Postovulatory stage (I), Hydrated ovarian follicles (diameter: 1.8 mm); oviducal eggs or corpora

REPRODUCTION IN THE SOUTH AFRICAN LACERTID PEDIOPLANIS BURCHELLI length (TBL, SVL TL) or logSVL was used to determine their scaling in cases where SSD existed. ANCOVA was used to compare the slopes of regression equations, which served as indicators of growth rate in the two sexes. The Mann-Whitney U-test was used to compare the incidence of broken tails between males and females. Differences between sets of data were considered signicant at P 0.05. All the statistical analyses were carried out using the software package STATISTICA 5.1 (StatSoft Inc.). RESULTS Spermatogenesis and the Testicular Cycle.In males, mean SVL was 53.4 6 3.3 mm (range: 45.661.5 mm; N 5 75); SVL of the smallest reproductively mature specimen was 45.6 mm. All testes appeared quiescent from February through August (late summer through late winter; Table 1, Stages 1 and 2). Although the rst sign of testicular recrudescence (Stage 3) was seen in a specimen from July, it was only in specimens from August that testicular recrudescence was most prevalent. The rst specimen with testes showing full spermatogenesis (spermiogenesis, Stage 5) appeared in September (spring). From October through January (early through late summer), the testes of most specimens showed maximum spermatogenic activity (Stage 5). The testes of one specimen from November, however, showed declining spermatogenesis (Stage 6) whereas that of another was already quiescent (Stage 7). By January, the testes of more than half the specimens were quiescent (Stages 1 and 7). Signicant monthly variation was found in testis mass and length as well as in mean seminiferous tubule diameter (testis mass: ANOVA: F7;67 5 21.5, P , 0.001; testis length: F7;67 5 20.7, P , 0.001; seminiferous tubule diameter, F7;67 5 21.4, P , 0.001; Fig. 2B). All three variables peaked in October (Stage 5) and were then signicantly greater than those of all the other months except September (LSD: P , 0.05). Among the rest of the months, the variables did not differ signicantly (LSD: P . 0.05), and they were at their lowest between January and March (Stage 1). Epididymus diameter and mass, and epithelium height varied signicantly among the months (diameter, H 5 33.9, P , 0.01; mass, H 5 29.4, P , 0.001; epithelium height, H 5 31.5, P , 0.001). In October (Stage 6), all three variables were signicantly higher than those of the other months (Dunn: P , 0.05). The rest of the months showed no signicant differences (Dunn: P . 0.05). Epididymus diameter was at its lowest in August (Stages 2 and 3), epithelium height in January through March (Stage 1), and mass in June through August (Stages 2 and 3).

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TABLE 1. Seasonal changes in spermatogenic activity in Pedioplanis burchelli males according to the classication system of Licht (1967).
Spermatogenic stages with number of lizards in each stage Month N 1 2 3 4 5 6 7

JAN FEB MAR APR MAY JUN JUL AUG SEP OCT NOV DEC

6 6 6 3 7 7 7 8 7 6 6 6

3 6 6 3 7 7 6 4

1 4 3

1 6 4 6

Oogenesis, the Follicular Cycle and Fecundity. In females, mean SVL was 54.1 6 3.8 mm (range: 45.262.9 mm; N 5 91) and the size of the smallest sexually mature specimen was 45.2 mm. The ovaries were inactive, with only clear (hydrated) follicles, from March through August (Stage 1). From September (Stage 2) to December (Stage 6) yellowish follicles and signs of ovulation (corpora lutea and oviducal eggs) were evident. Ovarian and oviducal mass and follicle diameter (Fig. 2B) all varied signicantly among the months (ovarian mass, H 5 26.9, P , 0.001; oviducal mass, H 5 27.3, P , 0.001; follicle diameter, H 5 26.7, P , 0.01). Ovarian and oviducal mass peaked in October and were then signicantly greater than that of the other months (Dunn: P , 0.05). Follicle diameter peaked in December and was signicantly greater than that of the other months (Dunn: P , 0.05) all of which did not differ signicantly from each other (Dunn: P . 0.05). Vitellogenic follicles were rst observed in September (spring), numbered 47 in 78 females (mean: 5.5 6 1.3) and their number was not correlated with SVL (rs 5 0.005, P . 0.05). Oviducal eggs were found simultaneously in both oviducts and were present from November through February. Clutch size ranged from 46 eggs per female (mean: 4.5 6 1.4) and was also not correlated with SVL (rs 5 0.01, P . 0.05). In November, a female with both oviducal eggs and vitellogenic follicles was found, suggesting that at least some females produced two clutches per reproductive season. Climatic Correlates.Signicant negative correlations were found between ambient temperature and testis mass (rs 5 0.30, P , 0.01). Stepwise multiple regression analysis showed that photoperiod was the best predictor of seasonal variation in testis mass (testis mass 5 2.22 photoperiod

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FIG. 2. Seasonal variation in testis mass, testis length and seminiferous tubule diameter in male Pedioplanis burchelli (A); seasonal variation in ovarian and oviducal mass, and follicle diameter in female P. burchelli (B). The numbers above the standard error bars indicate sample size which was the same for all three variables in each sex.

REPRODUCTION IN THE SOUTH AFRICAN LACERTID PEDIOPLANIS BURCHELLI

477

TABLE 2. The table shows the regression slopes and regression coefcients of the log-transformed SVL, TL, and head and limb measurements of male and female Pedioplanis burchelli. r is the regression coefcient, Fratios and P-values refer to results of male versus female comparisons using ANCOVA, with logSVL as covariate (except in the case of logSVL which uses logTBL as covariate). The numbers in parentheses denotes sample size in the case of the head and limb measurements; M denotes males and F females.
Female Male N 5 53 N 5 63 (91) (75) Measurement

r M; F

SVL TL HDL HDH HDW FL HL

0.60 0.90 0.90 1.10 0.90 0.80 0.60

0.42 0.40 0.60 0.70 0.80 0.50 0.40

0.71; 0.53; 0.76; 0.42; 0.62; 0.49; 0.28;

0.43 12.1 0.001 0.17 31.2 0.0001 0.63 103.7 0.0001 0.62 38.2 0.0001 0.81 77.9 0.0001 0.48 45.9 0.0001 0.40 41.8 0.0001

FIG. 3. Log-transformed body and head dimensions regressed against logTBL and logSVL in male (solid lines and triangles) and female (broken lines and squares) Pedioplanis burchelli; SVL and TL (A); HDH, HDL, and HDW (B); FL and HL (C).

with sex as factor and logTBL as covariate nevertheless showed that SVL in females was signicantly greater than that of equal sized males. In the case of TL the FMR was 0.87: TL of males was 13% longer than females. Relative TL of males was also shown to be signicantly greater than that of females using ANCOVA with logSVL as covariate (Fig. 3A, Table 2). ANCOVA with logSVL as covariate also showed that in all the head and limb dimensions compared males were signicantly greater than females (Figs. 3B, C, Table 2). The mean absolute tail lengths of males and females differed signicantly as did the slope of the relationship between TL and SVL (heterogeneous slopes) and the mean asymptotic tail lengths (lizards with an SVL larger than 58 mm; Student t-test, N 5 6, t 5 3.52, 10 df, P , 0.05). Of the sample of 77 males, 31.2% had tails showing signs of damage and/or regeneration while in the sample of 91 females; the gure was 30.8%. A Mann-Whitney U-test showed that these differences were not signicant (U 5 65, P . 0.05). DISCUSSION Reproduction was seasonal in P. burchelli with male and female cycles well synchronized. Although testicular recrudescence commenced earlier (late winter) than ovarian recrudescence (spring) both reached peak activity in summer, presumably the mating period, a pattern characteristic of prenuptial lizard species from the temperate region (Fitch, 1970). Clutch size ranged from 46 eggs with at least some females producing more than one clutch per reproductive season. The pattern of reproduction in P. burchelli was similar to that described for the Namib

1.4 maximum ambient temperature, rs 5 0.85, P , 0.001). Ovarian mass was positively and signicantly correlated with ambient temperature, photoperiod and rainfall (ambient temperature: rs 5 0.34, P , 0.01; photoperiod: rs 5 0.47, P , 0.001; rainfall: rs 5 0.53, P , 0.001). Stepwise multiple regression analysis showed that rainfall was the best predictor of seasonal variation in ovarian mass (ovarian mass 5 0.89 rainfall 1.14 maximum ambient temperature, rs 5 0.53, P , 0.001). Sexual Size Dimorphism and Damaged Tail Incidence.The FMR of SVL was 1.01: females were slightly (1%) larger than males. ANCOVA

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W. T. NKOSI ET AL. in females and rainfall was found to be negative (e.g., in Cordylus polyzonus polyzonus in the Free State; Van Wyk, 1989); thus, the suggestions advanced above may not always hold. For P. burchelli, a further advantage of reproducing during the rainfall season (summer) is that the arrival of its hatchlings coincides with the period of high insect availability. The lack of a correlation between follicle number and clutch size and SVL in P. burchelli may simply reect a narrow size range of females. A similar situation found in Podacris milensis led Adamopolou and Valakos (2000) to argue that it was possibly indicative of egg volume and size having reached an evolutionary optimum. Clutch size and egg volume also showed no increase with increasing body size in Acanthodactylus scutellatus (Frankenberg and Werner, 1992). The signicantly longer SVL of female P. burchelli compared to equal sized males is probably an adaptation for increasing fecundity, several studies having reported a positive relationship between increasing female body size and clutch size in lizards (e.g., Flemming and van Wyk, 1992; Heideman, 1994). The signicantly longer tail of males, however, may simply be attributed to a longer tail base to accommodate the pair of hemipenes as suggested by the Morphological Constraint hypothesis of King (1989). However, when applying the latter hypothesis to lacertid lizards, Barbadillo and Bauwens (1997) found support for it in only one of nine species studied. The possibility that it may have evolved as a result of higher predation pressure seems unlikely because of the similarity in broken tail incidence between the sexes. However, if the relatively longer limbs of males afford them greater running speed than females their presumed greater success at outrunning predators may offset higher predation pressure resulting in a similar incidence of broken tails to females. Avery et al. (1987), for example, found that running speed in Lacerta vivipara increased as stride length and stride rate increased; stride length being a function of limb length (Hildebrand, 1982). These suggestions require follow-up studies in P. burchelli. Relatively larger head dimensions (width, length, and height) in males compared to females may have several explanations. It could be the result of sexual selection, if aggressive male-male encounters take place during reproduction (Reaney and Whiting, 2002) or if females preferentially mate with larger headed males (Vitt and Cooper, 1985). It may also be diet related with males consuming larger prey than females (intraspecic food niche divergence), thereby relaxing intersexual competition (Shine, 1989; Shine et al., 1996). However, in the absence of information on aspects such as the mating system and diet,

Desert lacertid Meroles cuneirostris by Goldberg and Robinson (1979). That species produces 14 eggs per reproductive season and may also lay a second clutch. A second Namib Desert lacertid, Meroles anchietae, however, exhibits continuous reproductive activity, that is, a typical tropical cycle, with spermatogenic regression occurring only briey in autumn and females producing 2 4 clutches annually (Goldberg and Robinson, 1979). The latter authors ascribed this continuous reproductive pattern in M. anchietae to a continuous supply of food resources. This species is omnivorous, supplementing its winter diet with windblown seeds when insect food is scarce. However, M. cuneirostris is strictly insectivorous and, thus, suffers a food resource scarcity during winter. When compared to other species in its geographic area the reproductive cycle of P. burchelli corresponds to that reported for Mabuya capensis by Flemming (1994) but differs from that reported for Cordylus giganteus by van Wyk (1991, 1995) and Pseudocordylus melanotus melanotus by Flemming (1993a,b). In the latter two species, spermatogenesis is postnuptial and reproductive activity of males and females asynchronous. These differences in reproductive cycles shown by species from the same geographic area support the suggestion of reproductive conservatism among lizards put forward by James and Shine (1985) and Van Wyk (1998). The negative correlation between testis mass and ambient temperature was unexpected because increasing ambient temperature has traditionally been implicated as one of the major factors regulating testicular activity in reptiles (Duvall et al., 1982; Licht 1984). In contrast to males, ovarian mass in females was positively correlated with all three environmental variables as was expected. That rainfall was the best predictor of ovarian mass uctuations in females may be related to the higher energy demands of egg production, as food resource (insects) quantity and quality are usually dependent on rainfall. A number of studies have in fact reported a positive relationship between precipitation and insect productivity (Parker and Pianka, 1975; Ferguson et al., 1990). The commencing of vitellogenic activity in autumn in the insectivorous Pseudocordylus capensis coincident with increasing rainfall (Van Wyk and Mouton, 1998) is consistent with the suggestion of high energy demands of egg production. Van Wyk (1991) found that female C. giganteus would defer reproduction if energy reserves were not adequate, whereas female Sceloporus virgatus provisioned with abundant moisture-rich food produced greater clutch masses and more eggs per clutch than nonprovisioned females of similar body size (Abell, 1999). There are nevertheless cases where the correlation between reproduction

REPRODUCTION IN THE SOUTH AFRICAN LACERTID PEDIOPLANIS BURCHELLI current data cannot address the latter two suggestions. The relatively larger head in males could also be the result of a faster growth rate compared to females, the general view being that females allocate relatively more energy to reproduction after reaching sexual maturity than to somatic characters such as head size. Acknowledgments.We gratefully acknowledge L. Mashinini for assistance in collecting the specimens for this study and thank the University of the Free State (Qwaqwa campus) for logistical support. Research funds received by NJLH from the South African National Research Foundation helped to defray expenses. Constructive suggestions by two anonymous reviewers greatly improved the quality of the manuscript.

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Journal of Herpetology, Vol. 38, No. 4, pp. 480486, 2004 Copyright 2004 Society for the Study of Amphibians and Reptiles

New Species of Physalaemus Fitzinger, 1826 from Southern Bahia, Brazil (Anura, Leptodactylidae)
CARLOS ALBERTO GONCALVES CRUZ1
AND

BRUNO V. S. PIMENTA2

Departamento de Vertebrados, Museu Nacional, Universidade Federal do Rio de Janeiro, Quinta da Boa Vista, 20940-040 Rio de Janeiro, R J, Brazil ABSTRACT.A new species of Physalaemus belonging to the Physalaemus cuvieri species group is described from southern Bahia, Brazil. It inhabits the borders of Atlantic Rain Forest fragments, natural forest clearings, and cacao plantations. The new species is characterized by its medium size, robust body, warty skin, presence of inguinal glands, one pair of dorsal tubercles on the sacral region, and an interorbital black transversal stripe generally covering a pair of tubercles. RESUMEN.Uma nova especie de Physalaemus pertencente ao grupo de Physalaemus cuvieri, e descrita do sul da Bahia, Brasil. A especie e encontrada nas bordas de fragmentos de Mata Atlantica, clareiras naturais e plantacoes de cacau. A especie nova e caracterizada pelo seu tamanho medio, corpo robusto, pele verrucosa e ~ presenca de glandulas inguinais, de um par de tuberculos dorsais na regiao sacral e de uma faixa preta transversal na regiao interorbital, geralmente cobrindo um par de tuberculos.

The Neotropical frog genus Physalaemus Fitzinger, 1826, currently contains 41 valid species (Frost, 2002; Caramaschi et al., 2003). Lynch (1970) recognized and diagnosed four species groups in the genus: Physalaemus biligonigerus group, Physalaemus pustulosus group, Physalaemus signifer group, and Physalaemus cuvieri
1 Corresponding Author. E-mail: cagcruz@uol. com.br 2 E-mail: brunopimenta@softhome.net

group. Two species, Physalaemus deimaticus and Physalaemus rupestris, are not assigned to any of these species groups (Sazima and Caramaschi, 1986; Caramaschi et al., 1991). The Physalaemus cuvieri group is the most diverse, including 20 species distributed throughout South America (Frost, 2002). The P. cuvieri group is characterized by small to moderate size (1439 mm SVL), slender to stocky build, smooth to warty skin, rst nger not longer than the second, presence of an inner tarsal tubercle, presence of small,

NEW SPECIES OF PHYSALAEMUS noncompressed metatarsal tubercles (except Physalaemus albifrons), absence or presence of small inguinal glands (large in Physalaemus aguirrei), absence of parotoid glands, and presence of maxillary and premaxillary teeth (Lynch, 1970). In the coastal region of eastern Brazil, this group is represented by eight species: Physalaemus aguirrei, Physalaemus albifrons, Physalaemus barrioi, Physalaemus cicada, Physalaemus cuvieri, Physalaemus jordanensis, Physalaemus kroyeri, and Physalaemus soaresi (Frost, 2002). Feio et al. (1999) pointed out that P. aguirrei, Physalaemus maximus, Physalaemus olfersii, and P. soaresi should compose a new species group, but they lacked supporting evidence. Recent surveys in the Atlantic Rain Forest of southern Bahia, Brazil, resulted in the collection of a new species of this group. Herein, we present the description of this new species, the sonogram of the advertisement call, and some notes on natural history and distribution. MATERIALS AND METHODS Comparisons with the new species were restricted to the forms of the P. cuvieri group inhabiting the eastern coastal region of Brazil, as based on the authors examination of specimens and on information given by Izecksohn (1965), Bokermann (1966a,b, 1967), Lynch (1970), and Heyer et al. (1990). Specimens used in the description or examined for comparisons are deposited in EI (Eugenio Izecksohn collection, deposited in Universidade Federal Rural do Rio de Janeiro, Seropedica, RJ, Brazil), MCN (Museu de Ciencias Naturais PUC Minas, Belo Hori zonte, MG, Brazil), MNRJ (Museu Nacional, Rio de Janeiro, RJ, Brazil), and MZUSP (Museu de Zoologia, Universidade de Sao Paulo, SP, Brazil). Specimens examined are listed in Appendix 1. Abbreviations used in the measurements of specimens are SVL (snoutvent length), HL (head length), HW (head width), ED (eye diameter), TD (tympanum diameter), UEW (upper eyelid width), IOD (interorbital distance), IND (internostril distance), END (eyenostril distance), THL (thigh length), TBL (tibia length), and FL (foot length, including tarsus). All measurements are in millimeters, and SVL, HL, HW, THL, TBL, and FL were measured with calipers, whereas the other measurements were made with an ocular micrometer. Specimens were sexed by the presence or absence of vocal sac. Vocalizations were recorded with a Sony DAT TCD-D8 digital tape recorder with a Sennheiser K6/ME66 microphone set. Sonograms were produced by the software Avisoft-SASLab Light for Windows, version 3.74, using 16 bit resolution, 22 kHz sampling frequency, FFT with 256 points, FlatTop window, 50% overlap, and 100% frame.

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FIG. 1. Physalaemus erikae sp. n., holotype (MNRJ 30349), dorsal view.

Physalaemus erikae sp. n. Figures 12 Holotype.MNRJ 30349, adult male (Fig. 1), collected at Fazenda Vista Bela (168369S, 398559W), Municipality of Guaratinga, State of Bahia, Brazil, by B. V. S. Pimenta, R. T. Moura, A. Paglia, and R. V. Lopes on 14 September, 2000. Paratypes.MNRJ 28981, adult female, collected by B. V. S. Pimenta on 13 March 2001, MNRJ 28982, adult female, collected by B. V. S. Pimenta on 21 June 2001, MNRJ 28983, adult male, collected by B. V. S. Pimenta on 12 July 2001, and MNRJ 28985, adult female, collected by B. V. S. Pimenta on 20 March 2002, at Reserva Particular do Patrimonio Natural (RPPN) Esta c o Veracruz (168239S, 398109W), Municipality of a Porto Seguro, State of Bahia, Brazil; MNRJ 28984, adult female, collected by R. V. Lopes on November, 2001 at Fazenda Princesa do Pajau (178109S, 398509W), Municipality of Itamaraju, State of Bahia, Brazil; MNRJ 3002829, MNRJ 30348, three adult females, collected by B. V. S. Pimenta and R. V. Lopes on 18 July 2000, MNRJ 30347, MCN 219899, MCN 2203, four adult females, collected by B. V. S. Pimenta and P. H. C. Cordeiro on 30 April 2000, at RPPN Serra do Teimoso (158099S, 398319W), Municipality of Jussari, State of Bahia, Brazil; MNRJ 3034346,

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C. A. G. CRUZ AND B. V. S. PIMENTA

FIG. 2. Physalaemus erikae sp. n., holotype (MNRJ 30349). (A) Dorsal and (B) lateral views of head (scale 5 5 mm); ventral views of (C) hand and (D) foot (scale 5 1 mm).

two adult males and two adult females, collected on 12 and 14 September 2000, at the same locality of the holotype. Diagnosis.A species belonging to the P. cuvieri group, characterized by (1) medium size

(males 21.626.6 mm SVL, females 19.227.1 mm SVL); (2) body robust; (3) tympanum weakly distinct; (4) presence of inguinal glands; (5) dorsal skin texture warty or with longitudinal ridges; (6) presence of one pair of dorsal tubercles

NEW SPECIES OF PHYSALAEMUS on the sacral region; (7) presence of an interorbital black transversal stripe, generally covering a pair of tubercles; and (8) presence of a dorso-lateral dermal fold beginning at the posterior corner of the eye, ending near the inguinal region. Physalaemus erikae has a smaller size (SVL for males 21.626.6 mm) than P. albifrons, P. barrioi, P. cuvieri, and P. kroyeri (combined SVL for males 25.935.0 mm), warty skin (with longitudinal glandular lines in P. albifrons and P. barrioi, smooth or nely granular in P. cuvieri), and presence of inguinal glands (absent in these species). Physalaemus erikae is distinguished from P. aguirrei, P. cicada, P. jordanensis, and P. soaresi by its stocky build and warty skin (smooth in P. aguirrei, P. cicada, and P. soaresi; with longitudinal glandular lines in P. jordanensis). By its weakly distinct tympanum, P. erikae differs from P. barrioi, P. cicada, and P. cuvieri, which have indistinct tympanums. Physalaemus erikae is distinguished from P. aguirrei, P. barrioi, P. cicada, P. jordanensis, and P. soaresi by the absence of a black or darkbrown stripe covering the loreal region and anks and differs from P. jordanensis and P. barrioi by the lack of pigmentation on inguinal glands. Description of Holotype.Body robust; head wider than long; snout rounded in dorsal and lateral views (Fig. 2AB); nostrils ovoid, nonprotuberant, directed laterally; canthus rostralis distinct, straight; loreal region concave; eyes slightly protuberant; tympanum weakly distinct; supratympanic fold from posterior corner of eye to shoulder (Fig. 2B); a weakly marked dorsolateral fold beginning at posterior corner of eye, immediately above the supratympanic fold, ending near inguinal region; vocal sac subgular, large, expanded externally; choanae large, nearly round; tongue narrow, long; maxillary and premaxillary teeth visible under magnication. Arms slender, upper-arms shorter and moderately more robust than forearms; ngers slender, long; nger lengths I , II IV , III; brown nuptial asperities on thumbs; subarticular tubercles single, round; outer metacarpal tubercle medium, ovoid; inner metacarpal tubercle large, ovoid; few small supranumerary tubercles; ngers tip not expanded (Fig. 2C). Legs moderately robust; tibia longer than thigh; sum of tibia and thigh lengths smaller than SVL; presence of a discrete tarsal fold; toes slender, long, toe lengths I , II , V , III , IV; subarticular tubercles single, conical, protrunding; foot with an elliptical inner metatarsal tubercle, small; outer metatarsal tubercle small, conical, protrunding; few supranumerary tubercles, small; toes tip not expanded (Fig. 2D); tarsal tubercle on the distal third, round. Inguinal glands small, not pigmented; dorsal surfaces warty with scattered, weakly developed, short ridges; presence of one

483

conspicuous pair of dorsal black tubercles on the sacral region, and another pair between the eyes. Undersurfaces smooth, except for the thighs, which are nely areolate. Color in Preservative of Holotype.The general pattern is brown with small black blotches on dorsum and an interorbital black transversal stripe, covering a pair of tubercles. Dorsal surfaces of arms are light brown with a transverse black blotch; legs are light brown with dark brown transverse stripes and small black granules. Flanks are grayish-brown. Ventral surfaces have a cream background with chest and part of the belly gray marbled. Hands and feet are grayish-brown. Gular and cloacal regions are black. Measurements of Holotype.SVL 24.8; HL 7.2; HW 8.8; ED 2.6; TD 1.0; UEW 1.9; IOD 1.8; IND 1.7; END 2.7; THL 10.9; TBL 12.3; FL 13.9. Variation.In some specimens, the dorsolateral dermal fold is more evident. Arms in females are slender and the inner metacarpal tubercle is smaller than in males. Some specimens have a higher number of small supranumerary tubercles on hands. Instead of a warty skin with discrete, short ridges, females MCN 21982199, MNRJ 28982, and MNRJ 30348, and male MNRJ 30344 posses a pair of well-developed longitudinal ridges from upper eyelids to sacral region and another short pair between the eyes, and many longitudinal rows of small granules. A few specimens posses a less evident interorbital black transversal stripe or it is absent. Tarsal fold is not present in some specimens. Coloration pattern varies substantially among specimens. Gular region is cream with light-brown small blotches in females. Specimens MNRJ 3034430345 have a cream and light-brown general color pattern. Some specimens present a white or light-brown line on the urostylum. Females MNRJ 30347 and MCN 21982199 have yellowish-brown anks, and female MNRJ 28982 has a white vertebral line from snout to vent, interrupting the interorbital black transversal stripe. Variations on measurements are in Table 1. Vocalization.On 25 October 2001, two male P. erikae calling alternately were recorded emitting calls with one note consisting of eight harmonics between approximately 0.34 and 4.8 kHz, with descendent frequency modulation (Fig. 3A). The mean duration of the call was 0.56 sec (SD 5 0.01, range 5 0.530.57, N 5 16 vocalizations of two males), and the mean intercall interval (dened here as the time from the end of one call to the beginning of the next call) was 13.42 sec (SD 5 8.87, range 5 4.8535.92, N 5 16 vocalizations of two males). The fundamental harmonic was about 0.34 and 0.78 kHz; dominant frequency was 3.22 kHz, corresponding to the seventh

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C. A. G. CRUZ AND B. V. S. PIMENTA

TABLE 1. Mean (x), standard deviation (SD), and range of the measurements (in millimeters) of males and females of Physalaemus erikae sp. n.
Males (N 5 5) x SD Range x Females (N 5 13) SD Range

SVL HL HW ED TD UEW IOD IND END THL TBL FL

23.7 7.0 7.6 2.4 1.0 2.0 2.4 1.7 1.7 10.2 11.7 12.7

2.1 0.8 1.0 0.1 0 0.1 0.3 0.1 0.2 0.8 0.9 1.0

21.626.6 6.07.8 6.88.8 2.22.6 1.01.0 1.92.2 2.02.7 1.61.8 1.31.8 9.210.9 10.412.7 11.313.9

23.1 6.9 7.4 2.4 0.9 2.0 2.5 1.7 1.7 9.9 11.6 12.7

2.3 0.7 0.8 0.2 0.2 0.2 0.3 0.1 0.2 0.9 1.0 0.8

19.227.1 5.77.7 6.08.3 2.02.9 0.61.2 1.62.3 2.03.0 1.61.8 1.41.9 7.911.2 9.112.8 10.713.6

harmonic. The eighth harmonic ranged from 3.2 4.8 kHz. Physalaemus erikae call has a shorter duration than the calls of P. barrioi (1.21.4 sec; Bokermann, 1967), P. jordanensis (1.41.6 sec; Bokermann, 1967) and P. kroyeri (0.70.9 sec; Bokermann, 1966b), a higher dominant frequency

range than the calls of P. aguirrei (2.03.0 kHz; Bokermann, 1966a), P. albifrons (1.02.0 kHz; Bokermann, 1966b), P. cicada (2.03.0 kHz; Bokermann, 1966b), and P. cuvieri (0.580.8 kHz; Heyer et al., 1990). Information on the call of P. soaresi is not available. Habitat and Distribution.Specimens of P. erikae were found in temporary ponds in cow pastures at the borders of Atlantic Rain Forest remnants, natural clearings inside the forest, and cacao plantations. Males call from the edges of ponds or oating on shallow water. Females were found near ponds or dwelling on forest oor litter. Males also call during the day after heavy rains. This species was also registered by vocalization, with no specimens captured, at Fazenda Taquara (158589S, 398229W), Municipality of Belmonte, Fazenda Palmeira (158569S, 398389W), Municipality of Itapebi, Mata Cara Branca (168179S, 398259W), Municipality of Santa Cruz de Cabra lia, and Fazenda Alcoprado (178179S, 398409W), Municipality of Teixeira de Freitas. All these localities are also situated in southern Bahia, Brazil (Fig. 4). It is also expected to occur in northeastern Minas Gerais and northern Esprito Santo, because of the proximity and similarity of vegetation types between south Bahia and these regions. Etymology.The specic epithet erikae hon ors the second authors wife, Erika Costa Elias, in gratitude for her friendship and assistance.

FIG. 3. (A) Sonogram, (B) oscilogram, and (C) power spectrum of the advertisement call of Physalaemus erikae sp. n.; recorded at RPPN Estac o Veracruz, a Porto Seguro, Bahia, Brazil, on 25 October 2001, 2100 h. Air temperature 5 24.48C. Specimen not collected.

DISCUSSION The P. cuvieri species group, as proposed by Lynch (1970), shows a wide diversity of forms and life histories. The validity of this group as a natural group was questioned by Feio et al. (1999) when they suggested that P. aguirrei, P. maximus, P. olfersii, and P. soaresi could compose

NEW SPECIES OF PHYSALAEMUS

485

FIG. 4. Distribution of Physalaemus erikae sp. n. (dots) in southern Bahia, Brazil.

a separate species group, because of morphological similarities and the habit of reproducing on forested areas. It is possible that as the knowledge of the relationships between the Physalaemus species grows other groups will be proposed. The discovery of a new species of Physalaemus occurring over a relatively broad area demonstrates the current poor knowledge about taxonomy and composition of species in large regions of the Atlantic Rain Forest in Brazil. The last species of the P. cuvieri group described from Brazil was P. lisei, which occurs in the Atlantic Rain Forest of southern Brazil (Braun and Braun, 1977). Because of the high level of anthropogenic pressure that the Atlantic Rain Forest experiences, eld studies on distribution and composition of herpetological communities are vital. These studies might reveal other new species and identify centers of endemism, population uctuations, and other important data regarding the taxonomy, conservation, and management of lowland tropical forest anurans. Acknowledgments.D. L. Silvano made possi-

ble the collection of specimens in Bahia. We thank L. B. Nascimento and U. Caramaschi for comments on the manuscript; L. B. Nascimento also contributed with valuable suggestions and information; P. H. C. Cordeiro, R. T. Moura, A. Paglia, and R. V. Lopes for eld assistance; P. R. Nascimento for the drawings; the owners of the localities visited for the permission and support to collect in their areas; Conselho Nacional de Desenvolvimento Cient co e Tecnolo gico (CNPq) for the grants and nancial support. This study is a result of the subproject Ecological Approaches and Economic Instruments for the Establishment of the Discovery Corridor: a Strategy to Revert Forest Fragmentation of the Atlantic Forest at Southern Bahia, supported by the Project on the Conservation and Sustainable Use of the Brazilian Biological Diversity/Brazilian Ministry of the Enviroment (PROBIO/ MMA), Center for Applied Biodiversity Science/Conservation International (CABS/CI), The World Bank, and coordinated by the Institute for Social and Enviromental Studies of Southern Bahia (IESB).

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C. A. G. CRUZ AND B. V. S. PIMENTA LITERATURE CITED

IZECKSOHN, E. 1965. Uma nova especie de Physalaemus Fitzinger, do Estado do Rio de Janeiro (Amphibia, Anura). Revista Brasileira de Biologia 25:165168. LYNCH, J. D. 1970. Systematic of the American leptodactylid frog genera Engystomops, Eupemphix, and Physalaemus. Copeia 1970:488496. a SAZIMA, I., AND U. CARAMASCHI. 1986. Descric o de Physalaemus deimaticus, sp. n., e observacoes ~ sobre comportamento deimatico em P. nattereri (Steindn.)Anura, Leptodactylidae. Revista de Biologia 13:91101. Accepted: 9 July 2004.

BOKERMANN, W. C. A. 1966a. Dos nuevas especies de Physalaemus de Espiritu Santo, Brasil Physis 26:193 202. . 1966b. Notas sobre tres especies de Physalae mus de Maracas, Bahia (Amphibia, Leptodactyli dae). Revista Brasileira de Biologia 26:253259. . 1967. Tres novas especies de Physalaemus do sudeste brasileiro (Amphibia, Leptodactylidae). Revista Brasileira de Biologia 27:135143. BRAUN, P. C., AND C. A. S. BRAUN. 1977. Nova especie de Physalaemus do Rio Grande do Sul (Anura, Leptodactylidae). Revista Brasileira de Biologia 37: 86771. CARAMASCHI, U., L. C. CARCERELLI, AND R. N. FEIO. 1991. A new species of Physalaemus (Anura: Leptodactylidae) from Minas Gerais, southeastern Brazil. Herpetologica 47:148151. CARAMASCHI, U., R. N. FEIO, AND A. S. GUIMARAES-NETO. 2003. A new, brightly colored species of Physalaemus (Anura: Leptodactylidae) from Minas Gerais, southeastern Brazil. Herpetologica 59:519524. FEIO, R. N., J. P. POMBAL JR., AND U. CARAMASCHI. 1999. New Physalaemus (Anura: Leptodactylidae) from the Atlantic Forest of Minas Gerais, Brazil. Copeia 1999:141145. FROST, D. R. 2002. Amphibian Species of the World: an online reference. V2.21 (15 July 2002). Electronic database available at ,http://research.amnh.org/ herpetology/amphibia/index.html.. HEYER, W. R., A. S. RAND, C. A. G. CRUZ, O. L. PEIXOTO, AND C. E. NELSON. 1990. Frogs of Boraceia. Arquivos de Zoologia 31:231410.

APPENDIX 1
Specimens Examined Physalaemus aguirrei: MNRJ 2093820943 (Conceic o a da Barra, ES). Physalaemus albifrons: MNRJ 2406224063 (Brejo Santo, CE). Physalaemus barrioi: MZUSP 84821, MZUSP 84823, MZUSP 8428526, MZUSP 8482829 (topotypes, Sao Jose do Barreiro, SP). Physalaemus cicada: EI 6152 (paratopotype, Maracas, BA); MNRJ 28552, MNRJ 26040 (Brejo Santo, CE). Physalaemus cuvieri: MNRJ 66016615 (Fortaleza, CE). Physalaemus jordanensis: ZUEC 44794480, ZUEC 6257-6258 (Poc os de Caldas, MG) Physalaemus kroyeri: MZUSP 96515 96518, MZUSP 96522, MZUSP 96527, MZUSP 96544 96545, MZUSP 96472, MZUSP 96478, MZUSP 96483, MZUSP 96485, MZUSP 96487, MZUSP 96490, MZUSP 96493, MZUSP 96501 (Maracas, BA). Physalaemus soaresi: EI 17821783 (paratopotypes, Seropedica, RJ).