A molecular perspective on the phylogeny of the Hyla pulchella species group (Anura, Hylidae)

A molecular phylogenetic analysis of the Hyla pulchella species group was performed to test its monophyly, explore the interrelationships of its species, and evaluate the validity of the taxa that were considered subspecies of H. pulchella. Approximately 2.8 kb from the mitochondrial genes 12s, tRNA valine, 16s, and Cytochrome b were sequenced. The analysis included 50 terminals representing 10 of the 14-15 species currently recognized in the H. pulchella group, including samples from several localities for some taxa, several outgroups, as well as two species previously suspected to be related with the group (Hyla guentheri and Hyla hischoffi). The results show that the H. pulchella and Hyla circumdata groups are distantly related, and, therefore, should be recognized as separate groups. As currently defined, the H. pulchella group is paraphyletic with respect to the Hyla polytaenia group; therefore, we recognize the Hyla polytaenia clade in the H. pulchella group. Two subspecies of H. pulchella recognized by some authors are considered full species including Hyla pulchella riojana because it is only distantly related to H. pulchella, and Hyla pulchella cordobae because molecular and non-molecular evidence suggests that it is specifically distinct. With the inclusion of the H. polytaenia clade, H. guentheri, and H. bischoffi, and the recognition of the two former subspecies of H. pulchella as distinct species, the H. pulchella group now comprises 25 described species. All representatives of the H. pulchella group with an Andean distribution are monophyletic and nested within a clade from the Atlantic forest from south-southeastern Brazil/northeastern Argentina, and Cerrado gallery forest from central Brazil. (C) 2004 Elsevier B.V. All rights reserved.

CYTOGENETICS OF 4 BRAZILIAN HYLA SPECIES (AMPHIBIA-ANURA) AND DESCRIPTION OF A CASE WITH A SUPERNUMERARY CHROMOSOME

The chromosomes of Hyla fuscovaria, H. hayii and II. prasina, with 2n=24, and of Hyla sp. (aff. circumdata), a new species, with 2n=24 and 2n=25, were studied.The karyotypes with 2n=24 in the four species were very similar, with almost no differences in the size and morphology of the chromosomes. The numerical variability found in Hyla sp. (aff. circumdata) is due to the occurrence of a supernumerary chromosome in some specimens. NOR data obtained for the first time in the four species and C banding analysis of H. prasina indicate that such types of banding may be useful to differentiate species with very similar karyotypes, contributing to the understanding of chromosome evolution and the establishment of phylogenetic relationships among Brazilian Hyla species.

HYLA-LUCTUOSA, A NEW TREEFROG FROM SOUTHEASTERN BRAZIL (AMPHIBIA, HYLIDAE)

A new species of treefrog, Hyla luctuosa, is described from the Serra do Japi in southeastern Brazil. The new species is a member of the Hyla circumdata group characterized by large size, large tympanum, and rounded subarticular tubercles on the fingers. Descriptions of the tadpole and advertisement call and information on natural history are provided.

Chromosomal differentiation of Hyla nana and Hyla sanborni (Anura, Hylidae) with a description of NOR polymorphism in H-nana

Specimens of Hyla nana and Hyla sanborni from a syntopic population were studied cytogenetically. These species are morphologically very similar and are frequently misidentified, confused with each other. Both species had a diploid chromosome number, 2n = 30. However, the karyotypes of H. nana and H. sanborni differed considerably from each other in the number of submetacentric and telocentric chromosomes. The two species also differed in their primary NOR-bearing chromosomes (metacentric pair 13 in H. nana and telocentric pair 12 in H. sanborni). Additional nucleolus organizer regions (NORs) were detected by Ag-NOR staining and FISH in chromosome pairs 1, 5, 6, 12, and 14 in seven specimens of H. nana. Thus, a total of six patterns of NOR were identified. These differences in karyotype and in NOR location allowed the unambiguous identification of syntopic individuals of the two species. However, the chromosomal morphology of both species differed from that reported for populations from other geographic regions, suggesting that a systematic reevaluation of this group of Hyla may be necessary.

Tolerance to high temperatures in tadpoles of Leptodactylus fuscus and Hyla fuscovaria in temporary ponds (Amphibia, Leptodactylidae, Hylidae)

In temporary ponds tadpoles of the frogs Leptodactylus fuscus and Hyla fuscovaria may be exposed to temperatures up to 40-44°C. Experimental exposure to high temperature revealed survivals after 30 min at 42°C for H. fuscovaria and at 44°C for L. fuscus. -from Authors

Breeding activity of the neotropical treefrog Hyla elegans (Anura, Hylidae)

Breeding of Hyla elegans was monitored from August 1991 to July 1992 at a temporary pond in Ubatuba, State of Sao Paulo, southeastern Brazil. Males began to call as they entered the chorus, and defended their calling sites from other males, at times with physical interactions. Females, however, were not aggressive toward either males or other females. We found a positive correlation between the numbers of females and males in the chorus, but no significant correlation between OSR (number of reproducing females/number of reproducing males) and the number of males present. OSR was highly male-biased; on average, there were 10 males for each female; this low OSR may explain low average mating success of males. Females chose males as mates freely, and males did not attempt to intercept females approaching other males. Males in amplexus were larger and heavier than unmated, calling males. In addition, snoutvent lengths of males and females in amplexus were positively correlated, and males were, on average, 0.81 the length of females. Experimentally paired males and females with smaller or larger ratios of SVLs had a lower percent of fertilization than pairs near the population average.

Escalated aggressive behaviour and facultative parental care in the nest building gladiator frog, Hyla faber

Agonistic encounters and facultative parental care in Hyla faber were observed in two localities in southeastern Brazil. Maximum male density was 0.9 and 3.3 males/m2 in Campinas and Ribeirão Branco, respectively. Aggression was escalated and the highly variable aggressive calls were specific to each phase of the encounter. The last, more aggressive phases rarely occurred in Campinas; in Ribeirão Branco they occurred frequently. Male parental care (egg attendance) was common in Ribeirão Branco while it was never observed in Campinas. Egg attendance lasted one to two nights and was observed only during high male density. The main benefit of egg attendance seemed to be avoiding nest intrusion by other males (sunken eggs and/or embryos invariably die). Males may build additional nests during egg attendance, but attending males did not attract females (they did not call).

Comparing the tadpoles of Hyla geographica and Hyla semilineata

External morphology, internal oral anatomy, and chondrocranial anatomy were examined for tadpoles of Hyla geographica from the Amazon rainforest, Brazil, and Hyla semilineata from the Atlantic rainforest, Brazil. Here, we provide morphological larval data to help diagnose these closely related species. Scanning electron microscopy analysis of buccal morphology showed the most distinctive features between these species: the distance between the lingual papillae in the buccal floor of H. geographica is three times greater than that distance in H. semilineata, and the relative size of the lingual papillae in H. geographica is less than half their size in H. semilineata. Although the chondrocranium of both species is identical, and the external morphology of the larvae of both taxa is very similar, they differ greatly in size at most developmental stages. A multivariate analysis of covariance, corrected for stage and size, also showed a significant difference between morphometric measurements of the two species. These differences support the existence of two separate taxa.

Canto de anúncio de Hyla jimi e Hyla elianeae (Anura, Hylidae) na região de Botucatu, São Paulo, Brasil

In this work we present a description of the advertisement call of H. jimi, the locality type, and of H. elianeae, until now unknown in the literature. Field activities were carried out from August 1997 through June 1999; recordings were made in two open-area environments in the Botucatu region, São Paulo State. Vocalizations of 100 individuals (49 H. jimi and 51 H. elianeae) were recorded; nine characteristics of the advertisement call were examined. The advertisement calls of both species consist of consecutive series of simple notes with relatively fast repetition rates. Males of H. jimi and H. elianeae presented two patterns of note emission: one emitted by individuals beginning vocalization activity or isolated from the aggregate, and another emitted by males in chorus activity interacting with closely neighboring males. A significant difference was verified in the temporal structure of the two vocalization patterns.

Evaluating the karyotypic diversity in species of Hyla (Anura, Hylidae) with 2n = 30 chromosomes based on the analysis of ten species

Ten species of Hyla with 2n = 30 from Brazilian fauna were analysed cytogenetically. Hyla minuta is the unique presenting all bi-armed metacentric or submetacentric chromosomes in the karyotype, therefore, with the highest FN = 60. The remaining species have a variable number of uni-armed telocentric or subtelocentric chromosomes: H. cruzi, H. elianeae, and H. rubicundula with three pairs (FN = 54), H. berthalutzae, H. elegans, H. microps, and H. nana with four pairs (FN = 52), and H. nahdereri and H. sanborni with five pairs (FN = 50). The uni-armed elements are among pairs 5, 6, 7, 11, 14, and 15, which also appeared with metacentric or submetacentric morphology. The remaining chromosome pairs 1, 2, 3, 4, 8, 9, 10, 12, and 13 were never found to be telocentric or subtelocentric. AgNOR patterns are species-specific, the majority of the species exhibiting a single pair with AgNORs, with the exception of H. elegans and H. nana with more than one chromosome pair bearing this cytological marker. C banding was obtained in H. berthalutzae, H. cruzi, H. elegans, H. elianeae, H. microps, H. minuta, H. nahdereri, and H. nana, which showed positively stained centromeric heterochromatin. Our analysis confirms the great karyotypic diversity in the species of Hyla with 2n = 30, with no species sharing identical karyotypes.

Systematic review of the frog family Hylidae, with special reference to Hylinae: Phylogenetic analysis and taxonomic revision

Hylidae is a large family of American, Australopapuan, and temperate Eurasian treefrogs of approximately 870 known species, divided among four subfamilies. Although some groups of Hylidae have been addressed phylogenetically, a comprehensive phylogenetic analysis has never been presented. The first goal of this paper is to review the current state of hylid systematics. We focus on the very large subfamily Hylinae (590 species), evaluate the monophyly of named taxa, and examine the evidential basis of the existing taxonomy. The second objective is to perform a phylogenetic analysis using mostly DNA sequence data in order to (1) test the monophyly of the Hylidae; (2) determine its constituent taxa, with special attention to the genera and species groups which form the subfamily Hylinae, and c) propose a new, monophyletic taxonomy consistent with the hypothesized relationships. We present a phylogenetic analysis of hylid frogs based on 276 terminals, including 228 hylids and 48 outgroup taxa. Included are exemplars of all but 1 of the 41 genera of Hylidae (of all four nominal subfamilies) and 39 of the 41 currently recognized species groups of the species-rich genus Hyla. The included taxa allowed us to test the monophyly of 24 of the 35 nonmonotypic genera and 25 species groups of Hyla. The phylogenetic analysis includes approximately 5100 base pairs from four mitochondrial (12S, tRNA valine, 16S, and cytochrome b) and five nuclear genes (rhodopsin, tyrosinase, RAG-1, seventh in absentia, and 28S), and a small data set from foot musculature. Concurring with previous studies, the present analysis indicates that Hemiphractinae are not related to the other three hylid subfamilies. It is therefore removed from the family and tentatively considered a subfamily of the paraphyletic Leptodactylidae. Hylidae is now restricted to Hylinae, Pelodryadinae, and Phyllomedusinae. Our results support a sister-group relationship between Pelodryadinae and Phyllomedusinae, which together form the sister taxon of Hylinae. Agalychnis, Phyllomedusa, Litoria, Hyla, Osteocephalus, Phrynohyas, Ptychohyla, Scinax, Smilisca, and Trachycephalus are not monophyletic. Within Hyla, the H. albomarginata, H. albopunctata, H. arborea, H. boons, H. cinerea, H. eximia, H. geographica, H. granosa, H. microcephala, H. miotympanum, H. tuberculosa, and H. versicolor groups are also demonstrably nonmonophyletic. Hylinae is composed of four major clades. The first of these includes the Andean stream-breeding Hyla, Aplastodiscus, all Gladiator Frogs, and a Tepuian clade. The second clade is composed of the 30-chromosome Hyla, Lysapsus, Pseudis, Scarthyla, Scinax (including the H. uruguaya group), Sphaenorhynchus, and Xenohyla. The third major clade is composed of Nyctimantis, Phrynohyas, Phyllodytes, and all South American/West Indian casque-headed frogs: Aparasphenodon, Argenteohyla, Corythomantis, Osteocephalus, Osteopilus, Tepuihyla, and Trachycephalus. The fourth major clade is composed of most of the Middle American/Holarctic species groups of Hyla and the genera Acris, Anotheca, Duellmanohyla, Plectrohyla, Pseudacris, Ptychohyla, Pternohyla, Smilisca, and Triprion. A new monophyletic taxonomy mirroring these results is presented where Hylinae is divided into four tribes. Of the species currently in Hyla, 297 of the 353 species are placed in 15 genera; of these, 4 are currently recognized, 4 are resurrected names, and 7 are new. Hyla is restricted to H. femoralis and the H. arborea, H. cinerea, H. eximia, and H. versicolor groups, whose contents are redefined. Phrynohyas is placed in the synonymy of Trachycephalus, and Pternohyla is placed in the synonymy of Smilisca. The genus Dendropsophus is resurrected to include all former species of Hyla known or suspected to have 30 chromosomes. Exerodonta is resurrected to include the former Hyla sumichrasti group and some members of the former H. miotympanum group. Hyloscirtus is resurrected for the former Hyla armata, H. bogotensis, and H. larinopygion groups. Hypsiboas is resurrected to include several species groups - many of them redefined here - of Gladiator Frogs. The former Hyla albofrenata and H. albosignata complexes of the H. albomarginata group are included in Aplastodiscus. New generic names are erected for (1) Agalychnis calcarifer and A. craspedopus; (2) Osteocephalus langsdorffii; the (3) Hyla aromatica, (4) H. bromeliacia, (5) H. godmani, (6) H. mixomaculata, (7) H. taeniopus, (8) and H. tuberculosa groups; (9) the clade composed of the H. pictipes and H. pseudopuma groups; and (10) a clade composed of the H. circumdata, H. claresignata, H. martinsi, and H. pseudopseudis groups. Copyright © American Museum of Natural History 2005.

Calling sites and acoustic partitioning in species of the Hyla nana and rubicundula groups (Anura, Hylidae)

We analysed spatial and acoustic partitioning among four species of Hyla belonging to two species-groups: nana (H. nana and H. sanborni) and rubicundula (H. elianeae and H. jimi). Field activities were conducted at three permanent ponds, from 1998 through 2001. Four attributes of the calling sites were analysed: perch height, distance of the perch from the edge of the pond, type of perch (vegetation) and the individual's position on the perch. There was extensive overlap in the four calling-site variables analysed. However, we found spatial segregation did occur in calling site height and the distance of perches from pond edges. Bioacoustic analyses revealed behavioural differences among species in calling activity, both time of onset and peak calling in chorus. There was acoustic partitioning among species the fundamental frequency of the advertisement calls, principally as a function of the temporal structure (e.g. note duration, rate of note repetition, duration and rate of repetition of the calling pulses). We propose that differences in physical attributes of calling site and in characteristics of calls allow these species to exist in sympatry.

Variações morfológicas em Edessa rufomarginata e revalidação de E. albomarginata e E. marginalis (Heteroptera, Pentatomidae, Edessinae)

ABSTRACT: Based on the types of Edessa rufomarginata (De Geer, 1773) and its synonyms, on morphology of the paramere and coloration, seven patterns are described for E. rufomarginata. Pentatoma furcata Palisot de Beauvois, 1805, Cimex cruentus Fabricius, 1775, Aceratodes flavovirens Stål, 1855 and A. flavomarginatus Stål, 1855 are mantained as junior synonyms of E. rufomarginata. A. albomarginatus Stål, 1855 and A. marginalis Dallas, 1951 are removed from the synonymy of E. rufomarginata and are reinstated in Edessa. Aceratodes discolor Dallas, 1951 is removed from the synonymy of E. rufomarginata and is considered junior synonym of Edessa abdominalis Erichson, 1848.

The identity of Hyla leucotaenia Burmeister, 1861 (Anura: Hylidae)

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)