A Mata Atlântica é um dos ecossistemas mais severamente afetados pelos processos de perda e fragmentação de habitats, e, apesar dessa intensa ameaça, abriga ainda grande diversidade de anfíbios e répteis. O presente estudo teve como objetivo principal relacionar a diversidade de anuros e lagartos de serapilheira com o tamanho de fragmentos florestais, assim como com características ambientais dos fragmentos e da paisagem onde os mesmos estão inseridos. A amostragem da herpetofauna foi realizada em dois anos consecutivos durante a estação chuvosa numa paisagem fragmentada nos municípios de Capão Bonito e Ribeirão Grande, adjacentes ao Parque Estadual de Intervales e Fazenda Paraíso, utilizando a área da fazenda como controle, que representa a mata contínua. Os lagartos e anuros foram amostrados por meio de armadilhas de interceptação e queda (\"pitfall traps\") instaladas em 15 fragmentos e em seis localidades na área-controle. Além das características da estrutura e configuração da paisagem, como o tamanho dos fragmentos e a conectividade, outras variáveis ambientais que podem influenciar a distribuição e a abundância desses táxons também foram analisadas. A herpetofauna encontrada na região é típica de Floresta Ombrófila Densa, mas os resultados não esclareceram se a variação na composição da taxocenose de anuros entre as paisagens é uma resposta à continuidade ou à fragmentação da mata, ou se é uma resposta à distância geográfica entre as paisagens. De maneira geral, anfíbios anuros e lagartos de serapilheira são sensíveis à modificação de seus ambientes, porém a resposta a estas modificações é específica da espécie-, variando de acordo com aspectos particulares da história natural de cada espécie. A perda de habitat e a fragmentação florestal exercem efeitos negativos sobre algumas espécies de anfíbios e, conseqüentemente, sobre a diversidade local, sendo fatores importantes a serem considerados em estratégias de conservação e na formulação de diretrizes de conservação da diversidade biológica em paisagens fragmentadas do Planalto Atlântico de São Paulo / The Atlantic Forest is severely affected by processes of habitat loss and fragmentation, but despite this intense threat it hosts an enormous diversity of amphibians and reptiles. The main goal of the present study was to correlate the diversity of leaf-litter anurans and lizards to the size of forest fragments, as well as to the environmental characteristics of the fragments and of the landscape in which they are inserted. The study area is a fragmented landscape inserted in the municipalities of Capão Bonito and Ribeirão Grande, adjacent to Parque Estadual de Intervales and Fazenda Paraíso. Sampling of the herpetofauna was conducted during the rainy season in two consecutive years. Fazenda Paraíso was used as the control and denotes continuous forest. Anurans and lizards were collected with pitfall traps installed in 15 fragments and six sites inside the control-area. Besides the characteristics of landscape structure and configuration, like the size of the fragments and their connectivity, other environment variables that may affect the distribution and abundance of these taxa were also analysed. The herpetofauna found in the region is typical of tropical rainforests, although the results did not elucidate whether the variation in the composition of the anuran assemblage found among the landscapes is a response to the continuity or fragmentation of the forest, or if it is related to the geographical distance among the landscapes. Overall, leaf-litter anurans and lizards are sensible to habitat modification. The response to such modifications is, nevertheless, species-specific and varies according to characteristics of the natural history of each species. Habitat loss and forest fragmentation exert negative effects over some species of amphibians and ultimately over the local diversity and should thus be considered as important factors in conservation strategies and in the development of conservation guidelines in fragmented landscapes of the Atlantic Plateau of São Paulo.

Recognizing the mechanisms of speciation and the limits of species is essential to understand the origin of biodiversity and how to conserve it. The general aims of my investigations during my doctoral studies were two-fold: to study evolutionary patterns and processes, and to provide specific and superspecific taxonomic classifications that try to reflect evolutionary history. I have focused my research on anurans in their biodiversity hotspot, the American Tropics. I have used morphological, behavioral (mating calls), and genetic (DNA sequences) characters to study species boundaries between frogs of the genus Pristimantis and the family Centrolenidae (glassfrogs). The results show that the exclusive use of single lines of evidence or the application of arbitrary thresholds impair and bias our ability to recognize new species and limit the possibility to understand evolutionary processes. Only an integrative approach combining every source of evidence provides the necessary feedback to discover all species and test their identity by comparing independent sets of data. This approach further allows identifying those species that probably represent stable comparative units (well supported species hypotheses) and to flag taxa that require further assessment. Phylogenetic reconstructions based on seven nuclear and mitochondrial genes for about 100 species of glassfrogs revealed that previous hypotheses of relationships were mislead by rampant convergent evolution at the phenotypic level. None of the previously suggested classifications fit with the reconstructed evolutionary history. Consequently, we proposed a new classification consistent with this phylogeny. I also studied the tempo and mode of diversification among glassfrogs. Based on sequences from ten genes in 87 species, I estimated species divergence times, age-range correlation between sister species, and reconstructed ancestral areas and dispersal/vicariance events. The results revealed a complex model of diversification where geographical isolation seems to be the dominant scenario for speciation and only clades of altitudinal generalists have been able to spread across the Neotropical rainforests.

Made available in DSpace on 2014-06-11T19:35:44Z (GMT). No. of bitstreams: 0 Previous issue date: 2010-01-18Bitstream added on 2014-06-13T19:25:16Z : No. of bitstreams: 1 ramos_jzp_dr_rcla.pdf: 1486958 bytes, checksum: 1deaf8b552b3e70b44e3724804558867 (MD5) / Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) / Durante o período de fevereiro de 2007 a janeiro de 2009, foram estudadas as taxocenoses de anuros de quatro Municípios do Lagamar Paulista, litoral sul do Estado de São Paulo, um complexo estuarino-lagunar caracterizado por três ilhas continentais e porções continentais adjacentes a esta ilhas. Com o objetivo de inventariar as comunidades de anuros da costa (Iguape e Pariquera-Açu) e das ilhas (Ilha de Cananéia, Ilha do Cardoso e Ilha Comprida) e revelar padrões de diversidade e similaridade faunística, foram realizadas campanhas mensais com duração de três a quatro dias consecutivos nos quais as espécies de anuros foram registradas através do uso de duas metodologias complementares (armadilhas de interceptação e queda e procura ativa). Os resultados obtidos através da análise dos dados de composição comparada de espécies revelaram que as comunidades estudadas estão estruturadas. As comunidades das ilhas de Cananéia e Comprida apresentaram um alto grau de similaridade em diversidade de espécies. O mesmo pode ser dito para Pariquera-Açu em relação à Iguape. A análise molecular da estrutura de populações de cinco espécies de anuros ocorrentes nos cinco locais estudados [Rhinella ornata, Dendrophryniscus leucomystax, Hypsiboas albomarginatus, Scinax sp. 1 (aff. alter) e Haddadus binotatus] revelou padrões diferenciados de estrutura e diferentes cenários de formação destas populações. Mecanismos pretéritos relacionados à colonização das ilhas por espécies de anuros, bem como características intrínsecas das espécies colonizadoras podem ter sido os maiores responsáveis pelos padrões de estrutrura de comunidades e estruturas populacionais observados para os locais estudados. O presente estudo evidencia a importância de estudos multidisciplinares para a compreensão da estrutura e evolução de populações e comunidades de anuros. / From February 2007 to January 2009 we studied the anuran taxocenosis of four municipalities of an estuary-lagoon complex in the south coast of São Paulo State (Lagamar) composed by three continental Islands and continental areas adjacent to these Islands. In order to survey the anuran communities from the continental areas (Iguape Pariquera-Acu) and Islands (Cananéia, Cardoso, and Comprida), and to determine patterns of diversity and faunal similarity we conducted monthly surveys lasting three to four days each in which anuran species were registered through two complementary methods (pitfall traps with drift fences and active search). The species composition analysis revealed that the anuran communities of the Lagamar are structured. Anuran communities from Cananéia and Comprida Island, as well as Pariquera-Açu and Iguape, were more similar regarding species composition. The analysis of molecular diversity patterns of five species shared among the studied areas [Rhinella ornata, Dendrophryniscus leucomystax, Hypsiboas albomarginatus, Scinax sp. 1 (aff. alter), and Haddadus binotatus] revealed different structural patterns and historical scenarios of populations arrangements. Historical mechanisms related to island colonization by some anuran species as well as intrinsic characteristics of these species may account for contemporary patterns of community and population structure. The present study reinforces the importance of interdisciplinary studies for the comprehension of population and community structure and evolution.

Fundação de Amparo a Pesquisa no Estado do Rio Grande do Sul / Environmental changes resulting from climatic conditions may limit the reproductive activity of anurans. The calling activity is an important indicator of the temporal patterns of anuran activity, which may be regulated by several factors. Reproductive strategies, including the period of calling activity, time of tadpole and juvenile recruitment, and the larval rate of development, can be used for minimizing the adverse influence of environmental variables. Moreover, local environmental changes may generate disturbances (e.g., flooding, grazing, climatic events) and alter the availability of resources in the environment, which may lead to either a reduction or an increase in the local species diversity. In this context, the intermediate disturbance hypothesis (IDH) predicts that the species diversity would be higher at intermediate levels of disturbance. In freshwater systems, for instance, water bodies with intermediate hydroperiods (temporary ponds) would have higher species diversity because they would have fewer predators in relation to permanent ponds, and more resources availability in relation to ephemeral ponds. Thus, we aimed to determine the environment abiotic effects on the calling activity of an anuran community from the temperate wetlands of the extreme southern Brazil. We identified the reproductive patterns of species considering the periods of tadpole and juvenile recruitment, the larval development rate, and the influence of environmental variables on the species activity seasonality. We also established the disturbance frequency, duration and extent in water bodies, through the investigation of the relationship between the disturbance levels, the hydroperiod and the diversity of anurans. Data were collected over 17 months in coastal wetlands of the extreme portion of southern Brazil. Between December 2012 and April 2014 we registered 11 active species, distributed in the families Hylidae, Leptodactylidae, and Bufonidae. The calling activity of the community was positively related to water temperature and rainfall. The recruitment of tadpoles occurred during the rainy period, regardless of variation in the measured abiotic variables. The recruitment of juveniles also occurred during the rainy period and was related to increased water temperature. Our results indicate a link between the frequency and duration of the disturbance with the hydroperiod, as well as with tadpole richness and abundance. Overall, much of the species abiotic dependence was linked to the patterns of reproductive behavior and phylogeny; the occurrence of many species was also linked to the hydrological regime of water bodies. Also, our study demonstrates the direct effect of a disturbance event and the IDH on the studied anuran community, in which temporary water bodies presented a richest anuran community than ephemeral or permanent ones. / As variações ambientais decorrentes do clima representam um limitador para a atividade reprodutiva em anuros. A atividade de vocalização, por exemplo, é um importante indicativo do padrão de atividade diária e sazonal dos anuros e pode ser regulada por diversos fatores. As estratégias reprodutivas, tais como períodos de vocalização, época de recrutamento de girinos e juvenis, são utilizadas para amenizar a influência dessas variáveis. Adicionalmente, as variações climáticas podem refletir na forma de distúrbios sobre uma comunidade. Os distúrbios (e.g. alagamentos, pastejo, eventos climáticos) agem no ambiente alterando a disponibilidade de recursos, podendo causar o declínio ou aumento da diversidade de espécies. Nesse contexto a hipótese do distúrbio intermediário (HDI) prediz que em níveis intermediários de distúrbio a diversidade de espécies seria máxima. Em sistemas dulcícolas, por exemplo, corpos d água com hidroperíodos intermediários (temporários) apresentariam maior diversidade, por possuir menor número de predadores em relação aos permanentes e mais recursos disponíveis em relação aos efêmeros. Dessa forma, objetivamos averiguar os efeitos de fatores abióticos sobre a atividade de vocalização em uma comunidade de zona úmida temperada no extremo sul do Brasil. Identificamos os padrões reprodutivos das espécies, através do período de recrutamento de girinos e juvenis e tempo de desenvolvimento larval, bem como a influência de variáveis abióticas sobre a sazonalidade das espécies. Por fim, estabelecemos os atributos frequência, duração e extensão de distúrbio em corpos d água continentais, verificando a relação dos níveis de distúrbio com o hidroperíodo e diversidade de espécies de anuros. Os dados foram coletados ao longo de 17 meses em campos úmidos costeiros no extremo sul do Brasil. Foram registradas 11 espécies ativas de dezembro de 2012 a abril de 2014, distribuídas em três famílias: Hylidae, Leptodactylidae e Bufonidae. A atividade de vocalização sazonal da comunidade foi relacionada com a temperatura da água e pluviosidade, havendo respostas diferentes quando consideradas as espécies separadamente. O recrutamento de girinos ocorreu no período de chuvas, sendo independente das variáveis abióticas. O recrutamento de juvenis também ocorreu no período de chuvas e foi relacionado ao aumento da temperatura da água. Observamos, também, uma ligação entre os níveis dos atributos frequência e duração de distúrbio com o hidroperíodo, bem como com a abundância e riqueza de espécies de girinos presentes nesses ambientes. De uma forma geral, os resultados adquiridos mostram que grande parte da dependência abiótica apresentada pela atividade das espécies esteve ligada à filogenia e padrões comportamentais reprodutivos, onde a ocorrência de muitas espécies está ligada ao regime hídrico dos corpos d água. Por fim, os resultados encontrados no presente estudo demonstram a ação direta de um evento de distúrbio, bem como da HDI, sobre a comunidade de anuros, onde corpos d água temporários, com níveis intermediários de distúrbio, apresentam um maior número de espécies que efêmeros ou permanentes.

Made available in DSpace on 2014-06-11T19:30:15Z (GMT). No. of bitstreams: 0 Previous issue date: 2013-04-02Bitstream added on 2014-06-13T20:40:02Z : No. of bitstreams: 1 delgado_db_me_rcla.pdf: 856081 bytes, checksum: 0cd12b17e339c0315e379d1312fe09c7 (MD5) / Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) / Vocalizações são muito importantes para atração de fêmeas e delimitação de territórios em anuros de reprodução prolongada. Machos de Hypsiboas prasinus apresentam atividade de vocalização noturna ao longo de todo o ciclo anual na Serra do Japi, utilizando principalmente a vegetação marginal arbustiva e herbácea como sítios de vocalização. O padrão de atividade de vocalização dessa espécie foi acompanhado em um ciclo de 12 meses, de julho de 2011 a junho de 2012 e foram feitas gravações de suas vocalizações para análise bioacústica. Aqui são apresentados os parâmetros físicos de quatro das cinco vocalizações identificadas para esta espécie, sendo uma nota de anúncio introdutória (nota A), uma nota de anúncio utilizada em atividade de coro (nota B), uma nota de anúncio utilizada para retomada da atividade de vocalização (nota C) e uma nota agonística (nota D). Foi encontrada uma concentração de atividade de vocalização no mês de julho e não foi encontrada correlação significativa entre temperatura do ar e atividade de vocalização, sugerindo uma regulação da atividade baseada em interações comportamentais e não em limitações fisiológicas, conforme verificado por outros autores. Foi observado que alguns indivíduos passam a vocalizar a partir da superfície da água durante a estação fria e seca e aqui são discutidas as prováveis vantagens deste comportamento / Vocalizations have great importance for female attraction and territory delimitation in prolonged breeding anurans. Males of Hypsiboas prasinus present a year-round nocturnal calling activity, using mainly marginal herbaceous and bushy vegetation as calling sites. The temporal pattern of calling activity was monitored for a twelve-month period, from July 2011 to June 2012 and vocalizations were recorded for bioacoustics analysis. Herein are shown physical parameters for four of the five types of vocalizations found in this species, being one introductory advertisement call (note A), one chorus advertisement call (note B), one resuming activity advertisement call (note C) and one agonistic call (note D). A significant activity concentration was found in July and there was no significant correlation between air temperature and calling activity, suggesting a behaviorally-controlled calling pattern instead of a physiologically-controlled one, as observed by previous authors. During the cold dry season, some individuals were observed to shift their calling sites to the water surface and the plausible advantages of such behavior are discussed

A Mata Atlântica é um dos ecossistemas mais severamente afetados pelos processos de perda e fragmentação de habitats, e, apesar dessa intensa ameaça, abriga ainda grande diversidade de anfíbios e répteis. O presente estudo teve como objetivo principal relacionar a diversidade de anuros e lagartos de serapilheira com o tamanho de fragmentos florestais, assim como com características ambientais dos fragmentos e da paisagem onde os mesmos estão inseridos. A amostragem da herpetofauna foi realizada em dois anos consecutivos durante a estação chuvosa numa paisagem fragmentada nos municípios de Capão Bonito e Ribeirão Grande, adjacentes ao Parque Estadual de Intervales e Fazenda Paraíso, utilizando a área da fazenda como controle, que representa a mata contínua. Os lagartos e anuros foram amostrados por meio de armadilhas de interceptação e queda (\"pitfall traps\") instaladas em 15 fragmentos e em seis localidades na área-controle. Além das características da estrutura e configuração da paisagem, como o tamanho dos fragmentos e a conectividade, outras variáveis ambientais que podem influenciar a distribuição e a abundância desses táxons também foram analisadas. A herpetofauna encontrada na região é típica de Floresta Ombrófila Densa, mas os resultados não esclareceram se a variação na composição da taxocenose de anuros entre as paisagens é uma resposta à continuidade ou à fragmentação da mata, ou se é uma resposta à distância geográfica entre as paisagens. De maneira geral, anfíbios anuros e lagartos de serapilheira são sensíveis à modificação de seus ambientes, porém a resposta a estas modificações é específica da espécie-, variando de acordo com aspectos particulares da história natural de cada espécie. A perda de habitat e a fragmentação florestal exercem efeitos negativos sobre algumas espécies de anfíbios e, conseqüentemente, sobre a diversidade local, sendo fatores importantes a serem considerados em estratégias de conservação e na formulação de diretrizes de conservação da diversidade biológica em paisagens fragmentadas do Planalto Atlântico de São Paulo / The Atlantic Forest is severely affected by processes of habitat loss and fragmentation, but despite this intense threat it hosts an enormous diversity of amphibians and reptiles. The main goal of the present study was to correlate the diversity of leaf-litter anurans and lizards to the size of forest fragments, as well as to the environmental characteristics of the fragments and of the landscape in which they are inserted. The study area is a fragmented landscape inserted in the municipalities of Capão Bonito and Ribeirão Grande, adjacent to Parque Estadual de Intervales and Fazenda Paraíso. Sampling of the herpetofauna was conducted during the rainy season in two consecutive years. Fazenda Paraíso was used as the control and denotes continuous forest. Anurans and lizards were collected with pitfall traps installed in 15 fragments and six sites inside the control-area. Besides the characteristics of landscape structure and configuration, like the size of the fragments and their connectivity, other environment variables that may affect the distribution and abundance of these taxa were also analysed. The herpetofauna found in the region is typical of tropical rainforests, although the results did not elucidate whether the variation in the composition of the anuran assemblage found among the landscapes is a response to the continuity or fragmentation of the forest, or if it is related to the geographical distance among the landscapes. Overall, leaf-litter anurans and lizards are sensible to habitat modification. The response to such modifications is, nevertheless, species-specific and varies according to characteristics of the natural history of each species. Habitat loss and forest fragmentation exert negative effects over some species of amphibians and ultimately over the local diversity and should thus be considered as important factors in conservation strategies and in the development of conservation guidelines in fragmented landscapes of the Atlantic Plateau of São Paulo.

Orientadores: Shirlei Maria Recco-Pimentel, Carmen Sílvia Busin / Dissertação (mestrado) - Universidade Estadual de Campinas, Instituto de Biologia / Made available in DSpace on 2018-08-17T01:27:16Z (GMT). No. of bitstreams: 1 Bruschi_DanielPacheco_M.pdf: 3876863 bytes, checksum: 337679c24ea52e966f84be587a0058cb (MD5) Previous issue date: 2010 / Resumo: A taxonomia e as relações de parentesco em Phyllomedusa são temas de constantes discussões. Hipóteses sobre os relacionamentos intra- e interespecíficos desse gênero decorrem basicamente de análises morfológicas e comportamentais, o que não têm sido suficiente para responder alguns dos questionamentos. O grupo de P. hypochondrialis, o maior dentro do gênero, apresenta dificuldades na sua delimitação e, até o momento, não foi observada nenhuma sinapomorfia que possa reunir as espécies atualmente alocadas no grupo, de maneira que outras ferramentas podem ser elucidativas para resolução dessa problemática. O presente trabalho teve como objetivo utilizar dados citogenéticos e de sequências de DNA mitocondrial para o estudo de algumas das espécies do grupo de P. hypochondrialis. Todos os espécimes apresentaram o número diplóide 2n=26 e morfologia dos cromossomos bastante conservada, o que permitiu a inferência de homeologias cromossômicas. Exceto nas populações de Phyllomedusa sp. (aff. azurea) que apresentam o par 7 submetacêntrico, os cariótipos das demais populações analisadas foram constituídos por seis pares metacêntricos (1, 4, 8, 11-13), seis submetacêntricos (2, 3, 5, 6, 9, 10) e um subtelocêntrico (par 7). A diferença detectada no par 7 pode ser atribuída à presença de uma NOR nos braços curtos dos cromossomos 7 submetacêntricos. Pequenas variações na morfologia de alguns pares cromossômicos, incluindo a localização da NOR, foram observadas em P. rohdei de Ilhéus/BA em relação à descrita por outros autores, corroborando a hipótese da provável existência de espécies crípticas sob esse nome. O cariótipo de P. nordestina se diferenciou dos demais principalmente pela grande quantidade de heterocromatina e pela posição da NOR em 9p. A análise de morfologia externa relativa aos dois caracteres indicados para diagnose e separação de P. hypochondrialis e P. azurea, aplicados a todos os espécimes de diferentes localidades brasileiras, mostrou variações intrapopulacionais não acompanhadas de variações citogenéticas e moleculares. Esses dados apontam a necessidade de uma re-avaliação desses caracteres de diagnose e separação para essas espécies. A análise conjunta de dados citogenéticos e moleculares das populações permitiu identificar a população de Belterra/PA como P. hypochondrialis, portadora de NOR intersticial em 8p. Dados moleculares sugerem que as populações de Uberlândia (Minas Gerais), de São Luís, Bacabeira e Urbano Santos (Maranhão) e de Porto Nacional (Tocantins) possivelmente correspondam a um mesmo táxon, P. azurea. Nesse caso, a NOR em 7p nas populações ao Norte (Maranhão e Tocantins) e em 4p ao sudeste (Minas Gerais) corresponderia a variação interpopulacional. Na análise filogenética molecular, os haplótipos de Chapada dos Guimarães + Santa Terezinha (Mato Grosso) formaram um clado e as populações de Laranjal do Jari (Amapá) e de Prainha (Pará) formaram cada uma um ramo independente. Os dados dessas quatro populações (portadoras de NOR pericentromérica em 8q) sugerem que uma revisão minuciosa deva ser realizada para auxiliar no esclarecimento de seus status taxonômicos. / Abstract: Taxonomy and phylogenetic relationships of Phyllomedusa have been subject of continuous discussions. Hypotheses about the intra- and interspecific relationships within this genus have basically arised from morphological and behavioral characteristics, which have not been enough to elucidate the problems as the assigning of species to the P. hypochondrialis group. So, other tools may help to solve this problem. This work aimed to contribute with cytogenetic analysis and mitochondrial DNA sequencing data to the understanding of intra- and interspecific relationships involving the species P. rohdei, P. nordestina, P. hypochondrialis and P. azurea of the P. hypochondrialis group. These species showed the same chromosome number, 2n=26, with a very similar morphology. All populations had karyotypes with six metacentric (1, 4, 8, 11-13), six submetacentric (2, 3, 5, 6, 9, 10) pairs and one subtelocentric pair (7). The populations of Phyllomedusa sp. (aff. azurea) from São Luis, Bacabeira, Urbano Santos (Maranhão state) and Porto Nacional (Tocantins state) had the pair 7 submetacentric. This morphological difference in pair 7 can be attributed to the Nucleolus Organizer Region (NOR) that is located only in the submetacentric pair 7. The karyotype of P. nordestina was distinguished from P. rohdei by the large amount of heterochromatin and by the position of the NOR in chromosome 9p, whereas in P. rohdei it is located in 9q. The karyotype of P. rohdei also differed from that described by other author, suggesting the existence of cryptic species. Brazilian populations of P. azurea, P. hypochondrialis and other populations related to those species were analyzed. Intrapopulational variations in characters of external morphology were not associated with cytogenetic and molecular variations, showing that it is necessary a reevaluation of the diagnosis and separation characters for those species. The combined analysis of molecular and cytogenetic data allowed classifying the population of Belterra/PA as P. hypochondrialis, with NOR in pair 8p. Molecular data suggested that populations from Uberlândia (Minas Gerais), São Luiz, Bacabeira e Urbano Santos (Maranhão) and Porto Nacional (Tocantins) probably correspond to the taxon P. azurea. By comparing the karyotype of specimens from North populations (Maranhão and Tocantins) and from Southeast populations (Minas Gerais) different NOR positions were observed and interpreted as an interpopulational variation. The genetic diversity observed among populations with pericentromeric NOR in 8q (Laranjal do Jari, Prainha and Chapada dos Guimarães + Santa Terezinha; three clades in the molecular phylogenetic analysis) suggests that these populations could be in incipient process of speciation. / Mestrado / Biologia Celular / Mestre em Biologia Celular e Estrutural

A taxonomia produz uma importante contribuição para o conhecimento da biodiversidade. A taxonomia tradicional apresenta limitações quando a morfologia se mostra incapaz de revelar a diversidade dentro de grupos, necessitando a integração de diferentes tipos de dados. Espécies representam categoria básica de classificação biológica, existindo vários conceitos propostos. Um conceito unificado propõe que uma espécie representa um segmento de linhagem de uma metapopulação evolutivamente independente. Anuros apresentam ampla distribuição e são particularmente diversificados no Neotrópico. Sinais acústicos em anuros são amplamente estudados, sendo o mais abordado o canto de anúncio, por ser espécie-específico e ser útil no delineamento táxonômico entre populações/espécies. Oloylgon Fitzinger representa um grupo especioso de hilídeos e foi recentemente ressuscitado com base em análises moleculares para compor as espécies do antigo clado de Scinax catharinae. No presente estudo pretendemos contribuir com o conhecimento da diversidade de Ololygon, caracterizando morfologicamente e acusticamente populações e espécies, em particular: i) determinar a posição taxonômica de populações relacionadas à Ololygon centralis, provenientes do município de Araguari (Minas Gerais) e Cumari (Goiás); ii) re-caracterizar morfologia e cantos de três espécies (O. hiemalis, O. ranki e O. canastrensis) com o propósito de fornecer maiores detalhes sobre suas vocalizações e avaliar se o que está descrito na literatura corresponde com a nossa caracterização, e a partir disso, avaliar a diagnose entre O. hiemalis e O. ranki; iii) descrever pela primeira vez o repertório vocal de O. pombali da localidade tipo e reportar variações fenotípicas que não foram observadas antes. Espécimes depositados em museus foram avaliados quanto às suas morfologias e morfometrias. Gravações foram obtidas com gravadores profissionais/digitais e microfones direcionais acoplados, e analisadas no programa Raven Pro 1.5. As populações de Araguari e Cumari representam Ololygon centralis verdadeira, até que uma quantidade maior de evidências diga o contrário. O repertório vocal de Ololygon hiemalis é redescrito no presente estudo e, com isso, foi evidenciado que o canto tipo A (=anúncio) não foi reportado originalmente. Em relação à comparação entre O. hiemalis e O. ranki, foi considerado que elas são fenotipicamente indistinguíveis em termos de morfologia e vocalizações, mas apresentaram diferenças quantitativas em taxas de repetição de pulsos que podem ser atribuíveis a respostas comportamentais e/ou fisiológicas. O repertório vocal de O. canastrensis condiz com aquele descrito na literatura, apesar de haver algumas diferenças atribuíveis a variação intraespecífica. O repertório vocal de O. pombali é descrito pela primeira vez no presente estudo e existe uma variação em relação ao padrão dorsal de cor e manchas dessa espécie que não foi reportado originalmente, do qual não foi um caráter confiável para diagnose dessa espécie em relação aos demais congêneres. / Taxonomy yields an important contribution to the biodiversity knowledge. Traditional taxonomy is limited when morphology solely fails to reveal the real diversity within groups, requiring the integration of different kinds of data. Species represents the most basic category for biological classification, with currently a great array of concepts. A unified concept posits species as a lineage of a metapopulation that is evolutionarily independent. Anurans are widely distributed and are particularly diversified in the Neotropics. Acoustic signals in anurans are widely studied, being the most common one the advertisement call, as it has a species-specific characteristic and hence being useful in taxonomic delimitations between populations/species. Ololygon Fitzinger is a highly diversified hilydae genus and was recently resurrected on the basis of molecular analyses to shelter all species of the former Scinax catharinae clade. In the present study, we aim to contribute to the knowledge of Ololygon\"s diversity, by characterizing morphologically and acoustically species and populations, in particular: i) to determine the taxonomic position of populations closely related to Ololygon centralis, from the municipalities of Araguari (Minas Gerais) and Cumari (Goiás); ii) to re-characterize three species (O. hiemalis, O. ranki, and O. canastrensis) to provide further details on their vocalizations and evaluate if what is actually described in the literature corresponds to our descriptions, and also to evaluate the diagnosis between O. hiemalis and O. ranki; iii) to describe for the first time the vocal repertory of O. pombali from its type-locality and report phenotypic variations that were not previously reported. Specimens in museums were characterized regarding their morphology and morphometry. Recordings were acquired with digital/professional recorders and directional microphones coupled, and were analyzed in the software Raven Pro 1.5. Both populations from Araguari and Cumari represent O. centralis, until more evidences tell otherwise. The vocal repertory of O. hiemalis is redescribed in the present study, and with it, it was shown that the call type A (=advertisement) was not reported in the original description. When comparing O. hiemalis and O. ranki, it was considered that both species are phenotypically indistinguishable, although there are quantitative differences in pulse rate that is attributable to behavioral/physiological responses. The vocal repertory of O. canastrensis agrees with that described in the literature, despite of some differences attributable to intraspecific variation. The vocal repertory of O. pombali is described for the first time in the present study and this species show variation regarding dorsal pattern of color and blotches that was previously unreported, of which was not a reliable character to diagnose this species with respect to its congeners.

The Guiana Shield is a sub-region of Amazonia, one of the richest areas on earth in terms of species number. It is also one of the most pristine areas and is still largely unexplored. Species number, distribution, boundaries and their evolutionary histories remain at least unclear but most of the time largely unknown. This is the case for most Anurans, a group which is recognized as threatened globally and is disappearing even from pristine tropical forests. Given the pace of forest destruction and the growing concerns about climate change it is urgently necessary to obtain a better estimate of regional biodiversity in Amazonian frogs as well as a better understanding of the origin and distribution of Anuran diversity. Furthermore, given their sensitivity to climatic conditions, amphibians are a good model to investigate the influence of paleoclimatic events on Neotropical diversification which was supposedly the driving force on biotic evolution during Pleistocene in the Guiana Shield. I first test species boundaries in two species Scinax ruber and Rhinella margaritifera. These species are widely distributed, abundant and largely recognized as species complexes. I used an original species delineation method based on the combined use of mitochondrial and nuclear DNA in phylogenetic and phylogeographic analyses. Phylogenetic analyses demonstrated the polyphyly of Scinax ruber and Rhinella margaritifera. These species consist of multiple lineages that may all merit species status. Conflicting signals of mitochondrial and nuclear markers indicated the possibility of ongoing hybridization processes. Phylogeographic analyses added further information in support of the specific status of these lineages. Our results highlight the utility of combining phylogenetic and phylogeographic methods, as well as the use of both mitochondrial and nuclear markers within one study. This approach helped to better understand the evolutionary history of taxonomically complex groups of species. The assessment of the geographic distribution of genetic diversity in tropical amphibian communities can lead to conclusions that differ strongly from prior analyses based on the occurrence of currently recognized species alone. Such studies, therefore, hold the potential to contribute to a more objective assessment of amphibian conservation priorities in tropical areas. Subsequently, I tested if these first results on cryptic species are generalisable, questioning what would potentially be a minimum estimate of the number of cryptic frog species in Amazonia and the Guiana Shield, using mtDNA with multiple complementary approaches. I also combined isolation by distance, phylogenetic analyses, and comparison of molecular distances to evaluate threshold values for the identification of candidate species among these frogs. In most cases, geographically distant populations belong to genetically highly distinct lineages that could be considered as candidate new species. This was not universal among the taxa studied and thus widespread species of Neotropical frogs really do exist, contra to previous assumptions. Moreover, the many instances of paraphyly and the wide overlap between distributions of inter- and intra-specific distances reinforce the hypothesis that many cryptic species remain to be described. In our data set, pairwise genetic distances below 0.02 are strongly correlated with geographical distances. This correlation remains statistically significant until genetic distance is 0.05, with no such relation thereafter. This suggests that for higher genetic distances allopatric and sympatric cryptic species prevail. Based on our analyses, we propose a more inclusive pairwise genetic distance of 0.03 between taxa to target lineages that could correspond to candidate species. Using this approach, we identify 129 candidate species, two-fold greater than the 60 species included in the current study. This leads to estimates of around 170 to 460 frog taxa unrecognized in Amazonia-Guianas. As a consequence the global amphibian decline detected especially in the Neotropics may be worse than realised. The Rhinella margaritifera complex is characterisized by the presence of many cryptic species throughout its wide distribution, ranging from Panama to Bolivia and almost entire Amazonia. French Guiana has long been thought to harbor two species of this group, though molecular data analysed in previous chapters indicated as many as five lineages. I tested whether morphological measurements are correlated or not with genetic data using discriminant analysis and if diagnostic characteristics among the previously determined lineages can be used to describe these new species. This is a novel integrative method which can lead to a facilitation of the description of cryptic species that have been detected by phylogenetic and/or phylogeographic studies. These analyses, combined with published data of other Rhinella species, indicated that two of these lineages represent previously unnamed species. Two of the remaining are allocable to R. margaritifera while the status of the fifth is still unclear because so far it is morphologically indistinguishable from R. castaneotica. Determining if codistributed species responded to climate change in an independent or concerted manner is a basic objective of comparative phylogeography. Species boundaries, histories, ecologies and their geographical ranges are still to be explored in the Guiana Shield. According to the refugia hypothesis this region was supposed to host a forest refugium during climatic oscillations of the Pleistocene but the causes and timing for this have been criticized. We investigated patterns of genetic structure within 18 frog species in the eastern Guiana Shield to explore species boundaries and their evolutionary history. We used mtDNA and nuclear DNA and complementary methods to compare the genetic diversity spatially and temporally. With one exception all the species studied diversified repeatedly within the eastern Guiana Shield during the last 4 million years. Instead of one Pleistocene forest refugium the Guiana Shield has probably hosted multiple refugia during late Pliocene and Pleistocene. Most of these Pleistocene refugia were probably situated on the coast of French Guiana, Amapà, Suriname and Guyana. This diversification likely resulted from forest fragmentation. Many species deserve taxonomic revisions and their ranges to be reconsidered. The local endemism of the Anuran fauna of the Guiana Shield is likely to be much higher and some areas consequently deserve more conservation efforts. Specifically I questioned whether major intraspecific diversification started before the Pleistocene and occurred within the Guiana Shield or ex situ. According to ecological characteristics of the species involved I will test different diversification hypotheses. The consequences on the diversity and the endemism of the Guiana Shield will be explored. My results demonstrate that we have been grossly underestimating local biological diversity in the Guiana Shield but also in Amazonia in general. The order of magnitude for potential species richness means that the eastern Guiana Shield hosts one of the richest frog fauna on earth. In most of the species studied high levels of mtDNA differentiation between populations call for a reassessment of the taxonomic status of what is being recognised as single species. Most species display deep divergence between eastern Guiana Shield populations and Amazonian ones. This emphasizes that the local endemism in the Guiana Shield of these zones is higher than previously recognized and must be prioritised elements taken into account in conservation planning. Nevertheless, a few other species appear widely distributed showing that widespread species do exist. This underlines the fact that some species have efficient dispersal abilities and that the frog fauna of the eastern Guiana Shield is a mixture of old Guianan endemic lineages that diversified in situ mostly during late Pliocene and Pleistocene and more recently exchanged lineages with the rest of Amazonia. Recognizing this strong historical component is necessary and timely for local conservation as these zones are likely to be irremediably modified in the near future.

The Guiana Shield is a sub-region of Amazonia, one of the richest areas on earth in terms of species number. It is also one of the most pristine areas and is still largely unexplored. Species number, distribution, boundaries and their evolutionary histories remain at least unclear but most of the time largely unknown. This is the case for most Anurans, a group which is recognized as threatened globally and is disappearing even from pristine tropical forests. Given the pace of forest destruction and the growing concerns about climate change it is urgently necessary to obtain a better estimate of regional biodiversity in Amazonian frogs as well as a better understanding of the origin and distribution of Anuran diversity. Furthermore, given their sensitivity to climatic conditions, amphibians are a good model to investigate the influence of paleoclimatic events on Neotropical diversification which was supposedly the driving force on biotic evolution during Pleistocene in the Guiana Shield. I first test species boundaries in two species Scinax ruber and Rhinella margaritifera. These species are widely distributed, abundant and largely recognized as species complexes. I used an original species delineation method based on the combined use of mitochondrial and nuclear DNA in phylogenetic and phylogeographic analyses. Phylogenetic analyses demonstrated the polyphyly of Scinax ruber and Rhinella margaritifera. These species consist of multiple lineages that may all merit species status. Conflicting signals of mitochondrial and nuclear markers indicated the possibility of ongoing hybridization processes. Phylogeographic analyses added further information in support of the specific status of these lineages. Our results highlight the utility of combining phylogenetic and phylogeographic methods, as well as the use of both mitochondrial and nuclear markers within one study. This approach helped to better understand the evolutionary history of taxonomically complex groups of species. The assessment of the geographic distribution of genetic diversity in tropical amphibian communities can lead to conclusions that differ strongly from prior analyses based on the occurrence of currently recognized species alone. Such studies, therefore, hold the potential to contribute to a more objective assessment of amphibian conservation priorities in tropical areas. Subsequently, I tested if these first results on cryptic species are generalisable, questioning what would potentially be a minimum estimate of the number of cryptic frog species in Amazonia and the Guiana Shield, using mtDNA with multiple complementary approaches. I also combined isolation by distance, phylogenetic analyses, and comparison of molecular distances to evaluate threshold values for the identification of candidate species among these frogs. In most cases, geographically distant populations belong to genetically highly distinct lineages that could be considered as candidate new species. This was not universal among the taxa studied and thus widespread species of Neotropical frogs really do exist, contra to previous assumptions. Moreover, the many instances of paraphyly and the wide overlap between distributions of inter- and intra-specific distances reinforce the hypothesis that many cryptic species remain to be described. In our data set, pairwise genetic distances below 0.02 are strongly correlated with geographical distances. This correlation remains statistically significant until genetic distance is 0.05, with no such relation thereafter. This suggests that for higher genetic distances allopatric and sympatric cryptic species prevail. Based on our analyses, we propose a more inclusive pairwise genetic distance of 0.03 between taxa to target lineages that could correspond to candidate species. Using this approach, we identify 129 candidate species, two-fold greater than the 60 species included in the current study. This leads to estimates of around 170 to 460 frog taxa unrecognized in Amazonia-Guianas. As a consequence the global amphibian decline detected especially in the Neotropics may be worse than realised. The Rhinella margaritifera complex is characterisized by the presence of many cryptic species throughout its wide distribution, ranging from Panama to Bolivia and almost entire Amazonia. French Guiana has long been thought to harbor two species of this group, though molecular data analysed in previous chapters indicated as many as five lineages. I tested whether morphological measurements are correlated or not with genetic data using discriminant analysis and if diagnostic characteristics among the previously determined lineages can be used to describe these new species. This is a novel integrative method which can lead to a facilitation of the description of cryptic species that have been detected by phylogenetic and/or phylogeographic studies. These analyses, combined with published data of other Rhinella species, indicated that two of these lineages represent previously unnamed species. Two of the remaining are allocable to R. margaritifera while the status of the fifth is still unclear because so far it is morphologically indistinguishable from R. castaneotica. Determining if codistributed species responded to climate change in an independent or concerted manner is a basic objective of comparative phylogeography. Species boundaries, histories, ecologies and their geographical ranges are still to be explored in the Guiana Shield. According to the refugia hypothesis this region was supposed to host a forest refugium during climatic oscillations of the Pleistocene but the causes and timing for this have been criticized. We investigated patterns of genetic structure within 18 frog species in the eastern Guiana Shield to explore species boundaries and their evolutionary history. We used mtDNA and nuclear DNA and complementary methods to compare the genetic diversity spatially and temporally. With one exception all the species studied diversified repeatedly within the eastern Guiana Shield during the last 4 million years. Instead of one Pleistocene forest refugium the Guiana Shield has probably hosted multiple refugia during late Pliocene and Pleistocene. Most of these Pleistocene refugia were probably situated on the coast of French Guiana, Amapà, Suriname and Guyana. This diversification likely resulted from forest fragmentation. Many species deserve taxonomic revisions and their ranges to be reconsidered. The local endemism of the Anuran fauna of the Guiana Shield is likely to be much higher and some areas consequently deserve more conservation efforts. Specifically I questioned whether major intraspecific diversification started before the Pleistocene and occurred within the Guiana Shield or ex situ. According to ecological characteristics of the species involved I will test different diversification hypotheses. The consequences on the diversity and the endemism of the Guiana Shield will be explored. My results demonstrate that we have been grossly underestimating local biological diversity in the Guiana Shield but also in Amazonia in general. The order of magnitude for potential species richness means that the eastern Guiana Shield hosts one of the richest frog fauna on earth. In most of the species studied high levels of mtDNA differentiation between populations call for a reassessment of the taxonomic status of what is being recognised as single species. Most species display deep divergence between eastern Guiana Shield populations and Amazonian ones. This emphasizes that the local endemism in the Guiana Shield of these zones is higher than previously recognized and must be prioritised elements taken into account in conservation planning. Nevertheless, a few other species appear widely distributed showing that widespread species do exist. This underlines the fact that some species have efficient dispersal abilities and that the frog fauna of the eastern Guiana Shield is a mixture of old Guianan endemic lineages that diversified in situ mostly during late Pliocene and Pleistocene and more recently exchanged lineages with the rest of Amazonia. Recognizing this strong historical component is necessary and timely for local conservation as these zones are likely to be irremediably modified in the near future.