A new species of miniaturized toadlet, genus Brachycephalus (Anura: Brachycephalidae), from the atlantic forest of southeastern Brazil

A new species of miniaturized froglet (genus Brachycephalus) is described from Morro Prumirim in the municipality of Ubatuba, São Paulo, southeastern Brazil. Specimens were collected from the leaf litter at approximately 800 m above sea level. The new species is distinguished from all its congeners by the combination of the following characters: snout-vent length 8.7-13.4 mm; skin on head and dorsum with dermal ossification; general color orange, with a narrow brownish vertebral stripe and belly without spots; and skull, spinal processes of sacral and pre-sacral vertebrae, and process of the fourth vertebra hyperossified. Canonical variates analysis of linear body measurement demonstrates that the new species is easily discriminated from other Brachycephalus species with which it shares an intermediate condition of hyperossification. © 2012 by The Herpetologists League, Inc.

Correlações fisiológicas da carga parasitária em Rhinella icteriucus (Anura: marinus)

A presença de parasitas, que podem afetar inúmeros aspectos da biologia de seus hospedeiros, possivelmente levando a uma queda em seu desempenho. Entretanto, ao longo do estabelecimento da associação parasita-hospedeiro, alterações na fisiologia e morfologia deste último podem representar mecanismos compensatórios para os danos causados pelos parasitas. Sendo assim, carga parasitária deve ser um importante determinante do valor adaptativo (fitness) do hospedeiro. O objetivo do presente trabalho foi estabelecer possíveis correlações entre carga parasitária, massas de diversos órgãos e desempenho locomotor no sapo-cururu, Rhinella ictericus (Anura: Bufonidae). O desempenho locomotor dos indivíduos capturados de R. ictericus foi medido através do uso de uma pista circular de madeira. Durante um período de 10 minutos, os animais foram forçados a manter desempenho locomotor constante e a distância total percorrida foi corrigida para o comprimento rostro-cloacal. Depois das medidas de desempenho locomotor, os órgãos internos foram dissecados, tendo seus parasitas contabilizados. As massas de todos os órgãos escalaram positivamente com a massa corpórea. O número de parasitas nos pulmões e o índice corpóreo também se mostraram positivamente relacionados à variação em massa corpórea. Indivíduos com maior carga parasitária intestinal foram também aqueles que apresentaram um maior número total de parasitas, sugerindo a existência de variação individual na susceptibilidade a diferentes parasitas. O desempenho locomotor foi negativamente associado à carga parasitária total, demonstrando um efeito deletério causado pelos parasitas ao hospedeiro em um aspecto crucial de sua história de vida. Hospedeiros com maior carga parasitária pulmonar apresentaram maior massa intestinal e renal, sugerindo possíveis mecanismos compensatórios dos possíveis danos causados pelos parasitas

Advertisement call, colour variation, natural history, and geographic distribution of Proceratophrys caramaschii (Anura: Odontophrynidae)

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Diversidade genética em um grupo de anuros em miniatura endêmico da Mata Atlântica brasileira (Euparkerella Griffiths 1959).

A Mata Atlântica (MA) está entre as regiões com maior biodiversidade e mais ameaçadas do planeta. Esforços em diversas áreas do conhecimento têm sido feitos para que se tenha uma estimativa mais refinada da diversidade existente e sua organização ao longo do bioma. O crescente número de estudos que buscam reconstituir a história da diversificação da MA apontam para um cenário espacial e temporal complexo, havendo ainda uma lacuna no conhecimento dos processos em pequena escala. Vertebrados em miniatura têm se mostrado uma boa ferramenta para estudos de processos evolutivos em pequena escala. Assim, o gênero Euparkerella, endêmico de uma pequena região da MA dos Estados do Rio de Janeiro (RJ) e Espírito Santo (ES), foi escolhido como modelo para este estudo. No primeiro capítulo buscou-se descrever a diversidade existente dentro do gênero a partir de uma filogenia molecular. Para isso, utilizaram-se métodos bayesianos para gerar genealogias de genes e de espécies a partir de um fragmento de gene mitocondrial e quatro fragmentos de genes nucleares. Os resultados obtidos apontaram para uma grande diversidade críptica no gênero. Foram identificadas seis unidades evolutivas significativamente divergentes para o RJ: duas em Euparkerella cochranae, três em Euparkerella brasiliensis, e Euparkerella sp.. A espécie mais basal recuperada foi Euparkerella robusta, do ES, e estimou-se o início da diversificação do gênero para o final do Mioceno. O segundo capítulo descreve onze marcadores de microssatélites desenvolvidos para Euparkerella brasiliensis através do método de pirosequenciamento de nova geração 454. No terceiro capítulo estudou-se apenas uma unidade evolutiva, Euparkerella brasiliensis da área dos Três Picos/ RJ. A partir de marcadores de evolução rápida (microssatélites) e lenta (sequências de DNA) buscou-se compreender a estrutura e a dinâmica populacional desta unidade evolutiva em uma área bastante pequena (aprox. 20 km) sob influência de um gradiente ambiental altitudinal (40 m 1000 m). Foram identificadas, a partir dos microssatélites, duas subpopulações geneticamente distintas nas bordas do gradiente. O fluxo gênico se deu predominantemente das bordas para a zona de contato, onde foi observado o maior efetivo populacional. Tais resultados indicam que pequenas variações ambientais podem atuar no isolamento populacional em Euparkerella e corroboram o padrão de formas microendêmicas identificadas na filogenia. Futuros estudos devem ser feitos no sentido de buscar caracterizar morfologicamente as unidades evolutivas aqui identificadas; preencher as lacunas amostrais, especialmente no ES; e descrever os processos que atuam em pequena escala nas zonas de contato entre as unidades evolutivas e fatores limitantes a distribuição das mesmas.

锄足蟾总科两栖类系统演化研究

以线粒体DNA 的16S、ND4 和细胞色素B 三个基因片段的为测序和计算的 对象，对锄足蟾总科Pelobatidea 的角蟾科Megophryidae、锄足蟾科Pelobatidae、 掘足蟾科Scaphiopidae 和合跗蟾科Pelodytidae 这4 个科现生的所有属（除小臂蟾 属Leptobrachella 外）和绝大部分物种进行了分子系统学研究。综合的研究结果 表明，4 个科在系统演化上都各自形成独立的支系，证明了目前形态分类系统的 合理性。其亲缘关系的顺序依次是：东南亚的角蟾科与欧洲、西亚和非洲西北部 的锄足蟾科最近，然后二者与欧洲的另一个科合跗蟾科相聚，最后与北美洲的掘 足蟾科相聚。分化的先后为：掘足蟾科最先分化出来，然后是合跗蟾科，再后为 锄足蟾科，最后为角蟾科。角蟾科的两个亚科（角蟾亚科Megophryinae 和拟髭 蟾亚科Leptobrachiinae）在系统演化上也分别形成各自的支系，证明国内对亚科 的划分是合理的。这两个亚科虽然大部分为同域分布，但分别向不同的方向独立 演化。对属间系统演化关系的研究表明，锄足蟾科为单属、合跗蟾科为单属，掘 足蟾科为2 属（掘足蟾属Scaphiopus 和旱掘蟾属Spea），均为有效属，但角蟾科 原来划分的几个属的分类地位需要调整。本研究从整体上对这些属的分子系统演 化关系进行了分析和讨论，并对各主要科属的种间分子系统演化关系进行了分 析。通过本次实验, 我在角蟾亚科的属种分化和拟髭蟾亚科各属的属间和属内系 统演化关系的研究方面，提出了一些新的认识。 上述该分子系统研究还参照了形态学系统发育的研究和核型的比较研究。并 根据地质、地理、气候和生态环境及物种习性的有关资料对该类群的系统演化过 程和机理进行了分析和讨论。

棘蛙族(Tribe Paini)精子形态结构及精巢年周期研究

棘蛙族(Tribe Paini)隶两栖纲(Amphibia)、无尾目(Anura)、蛙科(Ranidae)、叉舌蛙亚科(Dicroglossinae)，由棘蛙属(Paa)、倭蛙属(Nanorana) 和沙巴蛙属(Chaparana)构成（Dubois,1992）。由于特殊的形态特征和染色体核型，棘蛙族受到国内外学者的广泛重视和研究，但是到目前为止，棘蛙族的系统发育关系尚未明晰，族下属种的分类和归属问题还有待进一步研究和新的证据出现。本文通过光学显微镜、电子显微镜和石蜡切片对棘蛙族10 物种的精子和精巢进行研究，旨在了解棘蛙族精子的形态、量度、超微结构特征及不同季节精巢结构的变化规律，同时为棘蛙族的系统研究提供新的依据，也为棘蛙族濒危物种的保护和经济物种的繁殖提供基础资料。研究结果表明：棘蛙族各属物种精子的形态基本相似，精子整体呈线形，由头部、中片和尾部构成。精子头部呈长条状，顶体呈锥状，位于头部顶端并向前伸出，中片较长，尾部波动弯曲。棘蛙族各属物种精子量度差异较大，将各属物种精子头部、中片、尾部、头宽、尾宽的量度数据进行聚类分析，结果表明棘蛙族10 物种可分为三类：第一类包括棘侧蛙、合江棘蛙、小棘蛙、棘腹蛙和棘胸蛙，特点是精子较短，全长在72.6～103.35µm 之间；第二类包括倭蛙、高山倭蛙、腹斑倭蛙，特点是精子较长，全长在107.74～129.75µm 之间；第三类包括隆肛蛙和双团棘胸蛙，特点是精子最长，全长在145.89～165.84µm 之间。棘蛙族各属精子超微结构基本相似：精子头部由顶体、细胞核构成；中片由中心粒、线粒体构成；尾部由单根轴丝构成。精子顶体横切呈圆环状，细胞核电子密度高；线粒体为卵圆形，呈环状围绕轴丝排列，线粒体数目较多，约30层；尾部轴丝为典型的9＋2结构，即由2根中央微管和9对外周微管组成。不同季节的倭蛙精巢结构变化表明倭蛙精巢每年只有一个生精周期，生精周期始于7 月，繁殖季节从5 月到6 月，生精高峰期为9 月；根据倭蛙不同季节精巢结构的变化，可将生精周期分为3 个阶段：第一阶段从7 月到9 月，为精子形成期；第二阶段从10 月到翌年4 月，为精子的贮存阶段，也即倭蛙的冬眠期；第三阶段从5 月到6 月，为精子的排放阶段，即倭蛙的繁殖期。不同季节的隆肛蛙精巢结构变化表明5 月为隆肛蛙的繁殖高峰期。根据棘蛙族各属精子的形态、量度和超微结构特征，结合已有的棘蛙族形态学、生态学、染色体核型及系统学研究成果，本文认为：1．基于精子数据对棘蛙族的划分和基于形态学及分子系统学数据对棘蛙族的划分均有相同之处，精子形态结构可为棘蛙族的系统研究提供新的证据。2． 棘蛙族各属精子的形态、量度及超微结构不仅与蛙科其他属种有明显差异，而且在无尾类中也较为特殊，精子学研究结果支持将棘蛙族从蛙科中分离出来，归隶于叉舌蛙科的叉舌蛙亚科的系统学修正。3. 精子的顶体、细胞核、中片的形态结构及量度可作为蛙科的分类指标。On the base of unique morphological and kyrotype characters, Dubois(1992)recognized three genera Paa, Narnorana, Chaparana as tribe Paini, which is amember of Dicroglossinae, Ranidae. In present study, the sperm shape, size andultrastructure of 10 paini species were investigated through the light and electronmicroscope, and testis structure of N. pleskei and F. quadrana was also studied. Wesuppose this study could offer some spermatological evidence to phylogeny andreproduction study of tribe Paini. The results were as follows:The sperm shape of tribe paini is homologically similar, the spermatozoa arefiliform, composed of elongate head, long mid-piece and waved tail. The acrosome isapically associated with the nucleus and extend anteriorly.The sperm length of tribe paini differ remarkably among genera. Cluster for thelength of sperm head, mid-piece, tail, total length, head-width, tail-width of ten painifrogs indicated the 10 species could be separated into three groups: GroupⅠcontainsP. shini, P. robertingeri, P. spinosa, P. exilispinosa, P. boulengeri, the spermatozoa ischaracterized with short in total length, ranging from 72.6µm to 103.35µm; GroupⅡcontains N. pleskei, N. parkeri, N. ventripunctata, the spermatozoa ischaracterized with relatively long in total length, ranging from 107.74µm to129.75µm; Group Ⅲ contains F. quadrana and P. yunnanensis, the spermatozoa is characterized with longest in total length, ranging from 145.89µm to 165.84µm. thethree groups based on spermatological data is partially match the classification basedon morphological and molecular data.The ultrastructure of spermatozoa in tribe paini is also basic similar, includingacrosome vescile, nuleus of the head proper, centriole, mitochondriol of themid-pieces, axoneme of the tail. The acrosome vescle is circle in TEM transversesection, the density of nucleus is high; The mitochondrions is oval, surrounding theaxial filament with about 30 layers of mitochondria; The axoneme has the typical 9+2pattern of microtubules.The seasonal changes in testis of N. pleskei indicates it has only onespermatogenesis circle, which begin in July, the reproduction season is from May toJune, the spermatogenesis is active in September. On the base of seasonal changes intestis, the spermatogenesis circle can be separated into three stages: In stageⅠfromJuly to September, spermatids are formed; In stage Ⅱ from October to April next year,the spermatozoa are stored in testis,which is the hibernated period; In stage Ⅲ fromMay to June, mature spermatozoa were released from the testis, which is thereproduction season of N. pleskei. As to F. quadrana, reproduction is active in May.With the previous study of morphology, ecology, karyotypes and phylogenyresearch of tribe Paini, the spermatological data in present study suggests:1. The spermatological classification of tribe paini is partially consistant with themorphological and molecular classification respectively.2.The sperm morphology and ultrustructure of tribe paini is unique not only inthe family Ranida but also in Anura, which suggest the tribe paini is monophyletic andmight be transfered from the family Ranida to the family Dicroglossidae based onmolecular evidence.3. The acrosome, nuleus, shape, length and ultrastructure of mid-piece can beused as an alternative taxonomic character in Anura.

The amphibians of São Paulo State, Brazil amphibians of São Paulo

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

Anfíbios do Estado de São Paulo, Brasil: conhecimento atual e perspectivas

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

Investigation of the cytocidal potential of Rhinella jimi skin methanol extracts

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

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.

Reinforcing and expanding the predictions of the disturbance vicariance hypothesis in Amazonian harlequin frogs: A molecular phylogenetic and climate envelope modelling approach

The disturbance vicariance hypothesis (DV) has been proposed to explain speciation in Amazonia, especially its edge regions, e. g. in eastern Guiana Shield harlequin frogs (Atelopus) which are suggested to have derived from a cool-adapted Andean ancestor. In concordance with DV predictions we studied that (i) these amphibians display a natural distribution gap in central Amazonia; (ii) east of this gap they constitute a monophyletic lineage which is nested within a pre-Andean/western clade; (iii) climate envelopes of Atelopus west and east of the distribution gap show some macroclimatic divergence due to a regional climate envelope shift; (iv) geographic distributions of climate envelopes of western and eastern Atelopus range into central Amazonia but with limited spatial overlap. We tested if presence and apparent absence data points of Atelopus were homogenously distributed with Ripley's K function. A molecular phylogeny (mitochondrial 16S rRNA gene) was reconstructed using Maximum Likelihood and Bayesian Inference to study if Guianan Atelopus constitute a clade nested within a larger genus phylogeny. We focused on climate envelope divergence and geographic distribution by computing climatic envelope models with MaxEnt based on macroscale bioclimatic parameters and testing them by using Schoener's index and modified Hellinger distance. We corroborated existing DV predictions and, for the first time, formulated new DV predictions aiming on species' climate envelope change. Our results suggest that cool-adapted Andean Atelopus ancestors had dispersed into the Amazon basin and further onto the eastern Guiana Shield where, under warm conditions, they were forced to change climate envelopes. © 2010 The Author(s).

Expression of acid phosphatase in the seminiferous epithelium of vertebrates

Acid phosphatases (AcPs) are known to provide phosphate to tissues that have high energy requirements, especially during development, growth and maturation. During spermatogenesis AcP activity is manifested in heterophagous lysosomes of Sertoli cells. This phagocytic function appears to be hormone-independent. We examined the expression pattern of AcP during the reproductive period of four species belonging to different vertebrate groups: Tilapia rendalli (Teleostei, Cichlidae), Dendropsophus minutus (Amphibia, Anura), Meriones unguiculatus (Mammalia, Rodentia), and Oryctolagus cuniculus (Mammalia, Lagomorpha). To demonstrate AcP activity, cryosections were processed for enzyme histochemistry by a modification of the method of Gömöri. AcP activity was similar in the testes of these four species. Testes of T. rendalli, D. minutus and M. unguiculatus showed an intense reaction in the Sertoli cell region. AcP activity was detected in the testes of D. minutus and O. cuniculus in seminiferous epithelium regions, where cells are found in more advanced stages of development. The seminiferous epithelium of all four species exhibited AcP activity, mainly in the cytoplasm of either Sertoli cells or germ cells. These findings reinforce the importance of AcP activity during the spermatogenesis process in vertebrates. © FUNPEC-RP.

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.

Caracterização do ciclo nucleolar e da formação do corpo cromatóide na espermatogênese de alguns vertebrados

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Carrying progeny on the back: reproduction in the Brazilian aquatic frog Pipa carvalhoi

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)