Apoptosis during the seasonal spermatogenic cycle of Rana catesbeiana

In the bullfrog Rana catesbeiana, testicular weight is constant throughout the year, but the volume densities of germinative and interstitial compartments undergo inverse changes from winter (non-breeding) to summer (breeding). The occurrence of apoptosis in the seminiferous lobules of bullfrogs was investigated in these two periods using sections stained with haematoxylin and eosin (H&E), the TUNEL (terminal deoxynucleotidyl transferase-mediated dUTP nick end labelling) method and transmission electron microscopy. TUNEL-positive cells were observed in the seminiferous lobules, and ultrastructural morphological details confirmed the occurrence of cell death by apoptosis. In summer, the occurrence of several spermatogenic processes (in addition to spermiogenesis and spermiation), and then the overconsumption of Sertoli cell-derived pro-survival factors, could be responsible for the increased density of apoptotic cells. Alternatively, the low apoptotic frequency in winter could be related to the constant homeostasis in the germinative compartment given that most lobules are filled with primary spermatocytes. As volume densities of interstitial and germinative compartments undergo inverse seasonal variations through the year, the incidence of apoptosis (in summer) could play a part in controlling the spermatogenic process, maintaining the lobular size when interstitial tissue is maximally developed. In winter, the low apoptotic cell density leads to spermatogenic recrudescence and, thereby, the production of an adequate quantity of spermatozoa for the next breeding period. Thus, apoptosis may participate not only in the maintenance of spermatogenic homeostasis, but also in the cyclical control of the different spermatogenic processes according to seasonal changes of the testicular compartments as a whole.

Water balance and locomotor performance in three species of neotropical toads that differ in geographical distribution

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

Seasonal metabolic changes in a year-round reproductively active subtropical tree-frog (Hypsiboas prasinus)

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

The evolution of jumping performance in anurans: morphological correlates and ecological implications

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

Ectopic mineralization of articular cartilage in the bullfrog Rana catesbeiana and its possible involvement in bone closure

Mineralization of the articular cartilage is a pathological condition associated with age and certain joint diseases in humans and other mammals. In this work, we describe a physiological process of articular cartilage mineralization in bullfrogs. Articular cartilage of the proximal and distal ends of the femur and of the proximal end of the tibia-fibula was studied in animals of different ages. Mineralization of the articular cartilage was detected in animals at 1 month post-transformation. This mineralization, which appeared before the hypertrophic cartilage showed any calcium deposition, began at a restricted site in the lateral expansion of the cartilage and then progressed to other areas of the epiphyseal cartilage. Mineralized structures were identified by von Kossa's staining and by in vivo incorporation of calcein green. Element analysis showed that calcium crystals consisted of poorly crystalline hydroxyapatite. Mineralized matrix was initially spherical structures that generally coalesced after a certain size to occupy larger areas of the cartilage. Alkaline phosphatase activity was detected at the plasma membrane of nearby chondrocytes and in extracellular matrix. Apoptosis was detected by the TUNEL (TDT-mediated dUTP-biotin nick end-labeling) reaction in some articular chondrocytes from mineralized areas. The area occupied by calcium crystals increased significantly in older animals, especially in areas under compression. Ultrastructural analyses showed clusters of needle-like crystals in the extracellular matrix around the chondrocytes and large blocks of mineralized matrix. In 4-year-old animals, some lamellar bone (containing bone marrow) occurred in the same area as articular cartilage mineralization. These results show that the articular cartilage of R. catesbeiana undergoes precocious and progressive mineralization that is apparently stimulated by compressive forces. We suggest that this mineralization is involved in the closure of bone extremities, since mineralization appears to precede the formation of a rudimentary secondary center of ossification in older animals.

Anurofauna em área de cerrado aberto no município de Borebi, estado de São Paulo, Sudeste do Brasil: uso do habitat, abundância e variação sazonal

O bioma Cerrado encontra-se descaracterizado e menos de três por cento de suas áreas originais está legalmente protegida. A anurofauna desse bioma não é muito rica quando comparado a outros biomas, porém há um grande número de espécies endêmicas. Aqui apresentamos uma lista de espécies de anuros registrados em uma lagoa em área de cerrado aberto do município de Borebi, região Centro-Oeste do estado de São Paulo, sudeste do Brasil. Durante 24 meses de estudo (2008 e 2009) caracterizamos a distribuição das espécies na lagoa estudada e descrevemos a variação sazonal das espécies. Foram registradas 27 espécies pertencentes a seis famílias: Bufonidae (duas espécies), Cycloramphidae (uma espécie), Hylidae (13 espécies), Leiuperidae (quatro espécies), Leptodactylidae (cinco espécies) e Microhylidae (duas espécies). A riqueza de espécies e abundância estiveram relacionadas com a precipitação. Dendropsophus minutus foi a espécie mais abundante e com registro de vocalização durante o ano inteiro. Rhinella ornata e Odontophrynus americanus foram restritas ao período seco e frio (abril a agosto). As outras espécies tiveram seu período de maior atividade nos meses chuvosos e quentes (setembro a março). A ocupação da lagoa variou com o tipo de vegetação e conforme a variação do seu volume de água, principalmente nos período de estiagem. A alta riqueza e abundância de anuros da lagoa pode ser resultado da ausência de peixes predadores, dos diversos tipos de microambientes do local e da ausência de outros corpos d'água próximos.

Predation of herps by spiders (Araneae) in the Brazilian Cerrado.

Two events of predation of herps by spiders in the Brazilian Cerrado are reported here. A lizard Micrablepharus atticolus (Sauria: Gymnophthalmidae) was found being preyed upon by Lycosa erythrognatha (Araneae: Lycosidae) and a frog Physalaemus cuvieri (Anura: Leiuperidae) was seen being preyed upon by Ancylometes sp. (Araneae: Ctenidae).

The inguinal macroglands of the frog Physalaemus nattereri (Leptodactylidae): structure, toxic secretion and relationship with deimatic behaviour

Amphibian skin is characterized by the presence of mucous glands, related to cutaneous breathing, reproduction and water balance, and granular glands, related to the production of toxins used in defence. In some species the granular glands can form accumulations in certain regions of the body. This is the case for inguinal macroglands of the leptodactylid frog Physalaemus nattereri, where these structures form a pair of black discs associated with deimatic behaviour. The morphology of the inguinal macroglands and their secretion were studied in this species and correlated to deimatic behaviour. The inguinal macroglands are formed from elongated granular glands that, in contrast with the granular glands of the rest of the skin, have small spherical granules with a proteinic content. In the dermis of the whole body, except for the inguinal macroglands and the inguinal region, a well-developed calcified dermal layer is observed. During deimatic behaviour these macroglands discourage a potential predator from attacking, but if visual cues are insufficient and the predator persists in the attack, atoxic secretion is eliminated in its mouth. This elimination is favoured by the absence of a calcified dermal layer in the macroglands, which makes the dermal region softer than the rest of the dorsal skin.

Diet and habitat use of two sympatric species of Philodryas (Colubridae), in south Brazil

Feeding habits and habitat use of the colubrid snakes Philodryas olfersii and P patagoniensis in southern Brazil are presented here. Philodryas olfersii and P patagoniensis are sympatric in the study area and both dwell in open and forested areas. Specimens preserved in collections and observations of snakes in the field yielded the data. Both species are diet generalists, feeding on small vertebrates, mainly frogs. Philodryas patagoniensis has a broader diet, a less variable frequency of food items, and fed on heavier prey than R olfersii. Seasonal variation in diet occurs in both species. The semi-arboreal Philodryas olfersii is more slender and has a longer tail than the terrestrial R patagoniensis, characters that may reflect differences in microhabitat use. There are a strong relationship between habitat use and frequency of a given food type. Differences in the use of food resources between P. olfersii and P patagoniensis seem to reflect differences in foraging microhabitats used by each species.

Habits hidden underground: a review on the reproduction of the Amphisbaenia with notes on four neotropical species

We review the information currently available on the reproduction of the Amphisbaenia and provide original data on the reproductive biology of four Neotropical species: Amphisbaena alba; A. mertensi; Cercolophia roberti, and Leposternon infraorbitale. In total, we compiled data for 22 species: 17 Amphisbaenidae, 1 Rhineuridae, 3 Bipedidae, and 1 Trogonophidae. The majority of the species were oviparous with the exceptions of Loveridgea ionidesii, Monopeltis anchietae, M. capensis, and Trogonophis wiegmanni. Viviparity was interpreted as a derived trait that evolved independently for at least 3 times within the Amphisbaenia. In most species, reproduction is synchronized with the hot and rainy season and seems to vary with latitude. Although Amphisbaenia eggs have been found in ant nests, it remains disputable whether this is an obligatory or even a preferable location for egg-laying. Incubation time in A. mertensii lasts 59 days and this is the first report encompassing egg-laying to hatching for any Amphisbaenia species. Nonetheless, a two months incubation period seems to be the common rule for oviparous Amphisbaenia. The general pattern of reproductive output in Amphisbaenia is characterized by a low number of eggs/embryos per clutch whose individual size is comparatively large in relation to adult body size. Eggs are markedly elongated on the long axis and arranged in-line within the abdominal cavity possibly to prevent/diminish biomechanic drawbacks of egg bearing. Hatchlings of A. mertensi possess an egg-tooth implanted at the upper jaw, exhibit positive geotropism, and display defensive behaviors known to be present in adults. Our review shows that our current knowledge of Amphisbaenia reproduction is fragmentary, often based on the examination of small samples, and heavily dependent on the publication of anedoctal observations. Future publications on this subject are encouraged.

Convergent evolution of aposematic coloration in Neotropical poison frogs: a molecular phylogenetic perspective

Poison frogs of the family Dendrobatidae contain cryptic as well as brightly colored, presumably aposematic species. The prevailing phylogenetic hypothesis assumes that the aposematic taxa form a monophyletic group while the cryptic species (Colostethus sensu lato) are basal and paraphyletic. Analysis of 86 dendrobatid sequences of a fragment of the 16S rRNA gene resulted in a much more complex scenario, with several clades that contained aposematic as well as cryptic taxa. Monophyly of the aposematic taxa was significantly rejected by SH-tests in an analysis with additional 12S and 16S rDNA fragments and reduced taxon sampling. The brightly colored Allobates femoralis and A. zaparo (Silverstone) comb. nov. (previously Epipedobates) belong in a clade with cryptic species of Colostethus. Additionally, Colostethus pratti was grouped with Epipedobates, and Colostethus bocagei with Cryptophyllobates. In several cases, the aposematic species have general distributions similar to those of their non-aposematic sister groups, indicating multiple instances of regional radiations in which some taxa independently acquired bright color. From a classificatory point of view, it is relevant that the type species of Minyobates, M. steyermarki, resulted as the sister group of the genus Dendrobates, and that species of Mannophryne and Nephelobates formed monophyletic clades, corroborating the validity of these genera. Leptodactylids of the genera Hylodes and Crossodactylus were not unambiguously identified as the sister group of the Dendrobatidae; these were monophyletic in all analyses and probably originated early in the radiation of Neotropical hyloid frogs.

Amphibians and reptiles from a highly diverse area of the Caatinga domain: composition and conservation implications

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

Anurans of a riparian forest in São Carlos, state of São Paulo, Brazil

No presente estudo foi realizado um inventário das espécies de anfíbios anuros que ocorrem em um fragmento de mata ciliar do Córrego do Espraiado, localizado na Universidade Federal de São Carlos (UFSCar), Município de São Carlos, estado de São Paulo, sudeste do Brasil. Durante os meses de março de 2009 a fevereiro de 2010 foram realizadas saídas de campo semanais nas quais foram registradas 13 espécies de anuros. A atividade reprodutiva das espécies de anuros componentes da comunidade estudada mostrou-se sazonal, sendo que houve correlação positiva e significativa entre o número de espécies em atividade reprodutiva e o fotoperíodo. Verificamos que a comunidade estudada mostrou-se mais semelhante a comunidades de anuros de florestas de transição entre Mata Atlântica e Cerrado do que a fragmentos de Cerrado próximos da região.

Vertebrados consumidos por Brycon opalinus (Pisces, Characidae) em rios do Parque Estadual da Serra do Mar, SP

A análise da alimentação da pirapitinga do sul (Brycon opalinus), peixe ameaçado de extinção de rios da Mata Atlântica da Serra do Mar na região Sudeste, revelou a ocorrência de itens alimentares incomuns. As espécies deste gênero são onívoras oportunistas e alimentam-se de itens vegetais e animais, tais como: flores, folhas, frutos e sementes e grande variedade de insetos. em três rios do Parque Estadual da Serra do Mar - Núcleo Santa Virgínia foram encontrados exemplares de B. opalinus que consumiram três itens animais incomuns, os anfíbios Hypsiboas aff. pardalis (Anura, Hylidae) e Eleutherodactylus guentheri (Anura, Leptodactylidae) e o mamífero Oligoryzomys cf. nigripes (Rodentia, Sigmodontinae). O registro do consumo destas espécies de vertebrados foi relacionado com o período de chuvas, quando o material animal ou vegetal carreado até o rio pode ser consumido por B. opalinus, mesmo que não sejam itens habituais para a espécie. A mata ripária preservada, como foi verificado nos três rios do Parque Estadual da Serra do Mar - Núcleo Santa Virgínia (SP), é de suma importância para o fornecimento de itens alimentares animais e vegetais e pela manutenção das condições bióticas e abióticas para a sobrevivência de B. opalinus.

Reproductive modes and fecundity of an assemblage of anuran amphibians in the Atlantic rainforest, Brazil

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