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.

Testicular morphological changes during the seasonal reproductive cycle in the bullfrog Rana catesbeiana

Spermatogenesis and steroidogenesis undergo seasonal variations during the reproductive cycle in amphibians. Testicular morphological and morphometric seasonal variations as well as interstitial lipidic inclusions and intralobular glycoconjugates were evaluated during seasonal cycle of Rana catesbeiana. Testes of frogs collected during the annual seasons were weighed for calculation of GSI (Gonadosomatic index). Seminiferous lobule diameters (DSL) and volume densities of seminiferous lobules (VvSL), excretory ducts (VvED), and interstitial tissue (VvIT) were analyzed. Semithin sections were submitted to Periodic Acid-Schiff (PAS) and Alcian Blue (AB) methods for detection of glycoconjugates, while lipidic inclusions were detected by Sudan Black B. GSI showed no significant variations during the year. Since VvED and VvIT increased significantly during summer and were inversely proportional to VvSL, a compensatory effect between the testicular compartments may be related to the maintenance of GSI. During autumn/winter, larger lobular diameters were observed in comparison to spring/summer when spermiogenesis and spermiation were commonly observed. The increased VvIT and the numerous lipidic inclusions in the interstitial cells during summer suggest a relationship between spermiogenesis and steroidogenesis. Besides the structural stability variations occurring in the IT and SL, a possible paracrine interaction between ED and IT should be also involved in the IT development during summer. The presence of PAS and AB-positive globular structures were observed in the seminiferous lobules and excretory ducts. These structures containing acid glycoconjugates appear to be Sertoli cell apical portions, which are accumulated in the lumen of the seminiferous lobules mainly during spermiation. © 2004 Wiley-Liss, Inc.

Acoustic repertoire and calling behavior of Scinax fuscomarginatus (Anura, Hylidae)

Vocalizations are important in anuran communication, especially during the breeding season. Calling affects spatial organization of males at breeding sites, promotes attraction of new individuals to the chorus, and attracts mature females for reproduction. We describe four distinct vocalizations (advertisement calls, sporadic calls, and two types of aggressive calls) emitted by adult males of Scinax fuscomarginatus. With the exception of the advertisement call, the remaining descriptions are novel. We also describe calling sites, calling season, and nightly calling activity. Fieldwork was made in a fragment of Cerrado (Estação Ecológica de Itirapina), Municipalities of Itirapina and Brotas, State of São Paulo, southeastern Brazil. At this ecological station, S. fuscomarginatus exhibits a complex acoustic repertoire, social interactions and a prolonged reproductive pattern similar to observations of congeneric species at other localities. Copyright 2005 Society for the Study of Amphibians and Reptiles.

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.

Cytogenetic and random amplified polymorphic DNA analysis of Leptodactylus species from rural and urban environments (Anura, Amphibia)

Cytogenetic and random amplified polymorphic DNA analyses carried out in the species Leptodactylus podicipinus, L. ocellatus, L. labyrinthicus, and L. fuscus from rural and urban habitats of the northwest region of São Paulo State, Brazil, showed that the karyotypes (2n = 22), constitutive heterochromatin distribution and nucleolus organizer region (NOR) location did not differ between the populations from the two environments. The in situ hybridization with an rDNA probe confirmed the location of the NORs on chromosome 8 revealing an in tandem duplication of that region in one of the chromosomes of L. fuscus. DAPI showed that part of the C-band-positive heterochromatin is rich in AT, including that in the proximity the NORs in L. podicipinus and L. ocellatus. The molecular analyses showed that the two populations (urban and rural) of L. podicipinus and L. fuscus are similar from a genetic point of view. The urban and rural populations of species L. ocellatus and L. labyrinthicus showed differences in genetic structures, probably due to urbanization which interferes with the dispersion of those frogs. The marked differences observed between the two populations of L. ocellatus can be representing the cryptic condition of the species. Unweighted pair-group method of analysis and genetic distance analysis detected the genetic proximity between L. ocellatus and L. fuscus. The results indicate that there was no reduction in the genetic diversity in the populations from the urban environment; however, the survival of these frogs would not be guaranteed in the case of an increase in human impact especially for populations of L. labyrinthicus and L. ocellatus. ©FUNPEC-RP.

Efeito da heterogeneidade ambiental na predação de girinos de Scinax fuscovarius e Physalaemus cuvieri por larvas de Odonata

The objective of this work were to analyze the effect of predation by Odonata naiads on two amphibian species with distinct habits - benthic and mid-water -and to verify whether the presence and architecture of macrophytes can mediate this interaction. All tadpoles and Odonata larvae were captured in a temporary pond. Sixteen tanks were used for three different treatments: Pistia, Salvinia and no macrophytes. Ten tadpoles of each species and two Odonata larvae were placed in each tank. The survival of tadpoles according to treatments was assessed through analysis of repeated measures. We concluded that the survival of P. cuvieri and S. fuscovarius tadpoles was not affected by the presence and architecture of the macrophytes (Pistia and Salvinia) or by their behavior.

Molecular phylogenetic relationships and phenotypic diversity in miniaturized toadlets, genus Brachycephalus (Amphibia: Anura: Brachycephalidae)

Toadlets of the genus Brachycephalus are endemic to the Atlantic rainforests of southeastern and southern Brazil. The 14 species currently described have snout-vent lengths less than 18. mm and are thought to have evolved through miniaturization: an evolutionary process leading to an extremely small adult body size. Here, we present the first comprehensive phylogenetic analysis for Brachycephalus, using a multilocus approach based on two nuclear (Rag-1 and Tyr) and three mitochondrial (Cyt b, 12S, and 16S rRNA) gene regions. Phylogenetic relationships were inferred using a partitioned Bayesian analysis of concatenated sequences and the hierarchical Bayesian method (BEST) that estimates species trees based on the multispecies coalescent model. Individual gene trees showed conflict and also varied in resolution. With the exception of the mitochondrial gene tree, no gene tree was completely resolved. The concatenated gene tree was completely resolved and is identical in topology and degree of statistical support to the individual mtDNA gene tree. On the other hand, the BEST species tree showed reduced significant node support relative to the concatenate tree and recovered a basal trichotomy, although some bipartitions were significantly supported at the tips of the species tree. Comparison of the log likelihoods for the concatenated and BEST trees suggests that the method implemented in BEST explains the multilocus data for Brachycephalus better than the Bayesian analysis of concatenated data. Landmark-based geometric morphometrics revealed marked variation in cranial shape between the species of Brachycephalus. In addition, a statistically significant association was demonstrated between variation in cranial shape and genetic distances estimated from the mtDNA and nuclear loci. Notably, B. ephippium and B. garbeana that are predicted to be sister-species in the individual and concatenated gene trees and the BEST species tree share an evolutionary novelty, the hyperossified dorsal plate. © 2011 Elsevier Inc.

Helminth parasites of hypsiboas prasinus (Anura: Hylidae) from two atlantic forest fragments, São Paulo State, Brazil

Seventy-seven males of Hypsiboas prasinus from 2 Atlantic forest fragments in the municipalities of Botucatu and Jundiaí, São Paulo State, Brazil, were examined for endoparasites. The frogs were captured in summer (January until March) and winter (July/August) of 2008 and 2009. Thirty-three males (75) from Botucatu were infected with Rhabdias cf. fuelleborni, cosmocerciid nematodes, and Cylindrotaenia americana. Twenty-five tree frogs (78.5) from Jundiaí were infected by Rhabdias cf. fuelleborni, Physaloptera sp., and cosmocerciid nematodes. Only cosmocerciid nematodes presented a statistically significance difference in prevalence (z 4.345; P < 0.001) and mean abundance (t 562.0; P < 0.001) between Botucatu and Jundiaí during the winter. Also, the cosmocerciids exhibited higher mean abundance (t 196.0; P 0.034) in winter when compared with summer at the Jundiaí site. Moreover, to our knowledge, this is the first report of C. americana in the Brazilian Hylidae. This study presents 4 new records of nematodes in H. prasinus. © 2012 American Society of Parasitologists.

Biochemical biomarkers in Scinax fuscovarius tadpoles exposed to a commercial formulation of the pesticide fipronil

One of the main pesticides used in the cultivation of sugarcane in São Paulo State, Brazil, is Regent®800WG, the main active compound of which is fipronil. Fipronil is a potent insecticide that eliminates pests, including insects resistant to pyrethroids, organophosphates (OP) and carbamates (CA). There is little known on the toxic effects of fipronil on non-target organisms, such as tadpoles of frogs. It is possible that this compound carries a high toxicity for these organisms, since the pesticide can be incorporated into aquatic environments during the rainy season, a time which coincides with the time of amphibian reproduction and the occurrence of tadpoles in the aquatic environment in this region. Thus, the pesticide could be contributing to the decline of amphibians in the northwest region of São Paulo state due to its wide use. This study aimed to test the influence of Regent®800WG on some biochemical systems of tadpoles (such as antioxidant defense systems) at different stages of development. The results of analysis from in vivo exposures demonstrated that only a few parameters in the groups exposed to fipronil responded to exposure to Regent®800WG, results which indicate that the pesticide instigates biochemical responses in tadpoles. Although catalase and glucose-6-phosphate dehydrogenase (G6PDH) were unchanged during the experiments, glutathione-S-transferase (GST) was inhibited in tadpoles, and the activity of glutathione reductase (GR) varied according to the exposure period and pesticide concentration. This data demonstrated the influence of the fipronil formulation on the metabolism of tadpoles, and showed that it can increase their susceptibility to environmental contaminants. © 2013 Elsevier Ltd. All rights reserved.

Systematics of spiny-backed treefrogs (Hylidae: Osteocephalus): An Amazonian puzzle

Spiny-backed tree frogs of the genus Osteocephalus are conspicuous components of the tropical wet forests of the Amazon and the Guiana Shield. Here, we revise the phylogenetic relationships of Osteocephalus and its sister group Tepuihyla, using up to 6134 bp of DNA sequences of nine mitochondrial and one nuclear gene for 338 specimens from eight countries and 218 localities, representing 89% of the 28 currently recognized nominal species. Our phylogenetic analyses reveal (i) the paraphyly of Osteocephalus with respect to Tepuihyla, (ii) the placement of 'Hyla' warreni as sister to Tepuihyla, (iii) the non-monophyly of several currently recognized species within Osteocephalus and (iv) the presence of low (<1%) and overlapping genetic distances among phenotypically well-characterized nominal species (e.g. O. taurinus and O. oophagus) for the 16S gene fragment used in amphibian DNA barcoding. We propose a new taxonomy, securing the monophyly of Osteocephalus and Tepuihyla by rearranging and redefining the content of both genera and also erect a new genus for the sister group of Osteocephalus. The colouration of newly metamorphosed individuals is proposed as a morphological synapomorphy for Osteocephalus. We recognize and define five monophyletic species groups within Osteocephalus, synonymize three species of Osteocephalus (O. germani, O. phasmatus and O. vilmae) and three species of Tepuihyla (T. celsae, T. galani and T. talbergae) and reallocate three species (Hyla helenae to Osteocephalus, O. exophthalmus to Tepuihyla and O. pearsoni to Dryaderces gen. n.). Furthermore, we flag nine putative new species (an increase to 138% of the current diversity). We conclude that species numbers are largely underestimated, with most hidden diversity centred on widespread and polymorphic nominal species. The evolutionary origin of breeding strategies within Osteocephalus is discussed in the light of this new phylogenetic hypothesis, and a novel type of amplexus (gular amplexus) is described. © 2013 The Norwegian Academy of Science and Letters.

Anurofauna em uma área de ecótono entre Cerrado e Floresta Estacional: diversidade, distribuição e a influência de características ambientais

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Diversidade de anfíbios anuros na Serra da Mantiqueira na região do Pico dos Marins

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Diversidade e estrutura de taxocenoses de anfíbios anuros em uma paisagem fragmentada no centro-oeste paulista

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Efeitos do canto de anúncio e do tamanho corporal no espaçamento entre machos em agregações de Dendropsophus nanus (Anura, Hylidae)

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Influência da heterogeneidade ambiental na diversidade, uso de hábitat e biocústica de anuros de área aberta no noroeste paulista

Pós-graduação em Biologia Animal - IBILCE