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The delimitation of evolutionarily significant units (ESUs) frequently results in controversy, but prioritising populations with evolutionary independence is essential for effective in situ conservation management. The American manatee (Trichechus manatus) is distributed along subtropical and tropical coastal waters from Florida (USA) to Alagoas (Brazil), and two subspecies are traditionally recognised, namely, T. m. latirostris, restricted to the Florida peninsula, and T. m. manatus, found in the remaining areas. However, this subspecific classification is not supported by genetic and morphologic evidence, which, rather, recognises two deeply differentiated populations or ESUs called Atlantic (Brazil) and Caribbean (from Venezuela to Florida). In this viewpoint paper, we compare both intraspecific divisions of T. manatus and the conservation implications. First, we used all available mtDNA evidence to test the genealogical clustering of the two American manatee ESUs by using a tree-based coalescent method. Second, we have used different models under a coalescent framework to estimate the historic gene flow among manatee populations. The analysis of the spatial distribution of mtDNA clusters confirmed the existence of the two suggested ESUs, rather than the two claimed subspecies. Furthermore, the best model to explain historic migration indicates that Brazilian manatees belong to an isolated population, whereas Florida and Caribbean populations are connected by more recent gene flow. These results have confirmed that T. manatus of the Caribbean, Gulf of Mexico and Florida belong to the same deme or Caribbean ESU, and the relatively isolated population inhabiting the Atlantic coast of Brazil belongs to the Atlantic ESU. Furthermore, both ESUs are separated by an interspecific hybrid zone (with the Amazonian manatee) located around the mouth of the Amazon River towards the Guianas coastline. The subdivision of two ESUs is also highly supported by karyotypic, morphological and ecological data, and is in clear disagreement with the traditional subspecies designations and the IUCN priorities, which manages Brazilian manatees as part of the Antillean manatee subspecies (T. m. manatus). Rather, Brazilian manatees should be considered as a full priority for conservation and require further taxonomic research; because of their deep history of isolation, they present high genetic and morphologic differentiation from all other American manatees.

Made available in DSpace on 2021-06-25T10:32:43Z (GMT). No. of bitstreams: 0 Previous issue date: 2021-06-01 Critical Ecosystem Partnership Fund The campo rupestre ecosystem is considered an old, climatically buffered, infertile landscape. As a consequence, long-term isolation is thought to have played an important role in the diversification of its biota. Here, we tested for hybridization between two endemic leaf frogs from the campo rupestre. We used sequence markers and coalescent models to verify haplotype sharing between the species, to test the existence and direction of gene flow, and to reconstruct the spatiotemporal dynamics of gene flow. Additionally, ecological niche modelling (ENM) was used to assess for potential co-occurrence by overlapping the climatic niche of these species since the middle Pleistocene. We found haplotype sharing and/or lack of differentiation in four nuclear fragments, one of them associated with introgression. The coalescent models support introgressive hybridization unidirectionally from Pithecopus megacephalus to P. ayeaye, occurring ∼300 kya. ENM corroborates this scenario, revealing areas of potential environmental niche overlap for the species at about 787 kya. These results contradict the expectation of reduced hybridization, while ENM suggests climatic fluctuation rather than stability for the two species. The reduced hybridization hypothesis needs to be further investigated because our results suggest that it may have unrealistic premises at least for animals. Departamento de Ciências Naturais Universidade Federal de São João del-Rei Programa de Pós-Graduação em Zoologia Departamento de Zoologia Universidade Federal de Minas Gerais Laboratório de Biodiversidade e Evolução Molecular Departamento de Genética Ecologia e Evolução Universidade Federal de Minas Gerais Laboratorio de Ecologia e Conservação Departamento de Botânica e Ecologia Instituto de Biociências Universidade Federal do Mato Grosso Programa de Pós-Graduação em Zoologia Instituto de Biociências Universidade Estadual Paulista (UNESP) Programa de Pós-Graduação em Genética e Biologia Molecular Departamento de Genética Evolução Microbiologia e Imunologia Universidade Estadual de Campinas Laboratório de Fauna e Unidades de Conservação Departamento de Engenharia Florestal Universidade de Brasília, Distrito Federal Programa de Pós-Graduação em Zoologia Instituto de Biociências Universidade Estadual Paulista (UNESP)

Made available in DSpace on 2019-10-04T12:34:45Z (GMT). No. of bitstreams: 0 Previous issue date: 2019-03-01 Fundação de Amparo à Pesquisa do Estado de Minas Gerais (FAPEMIG) Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) Even though Brazil is the world leader in amphibian diversity, a significant part of its richness remains unknown or hidden under cryptic taxa. Here, we used model-based species delimitation in an integrative taxonomic approach, by gathering molecular and morphometric data to assess cryptic taxa within the monkey frogs Pithecopus rohdei, from the Atlantic Forest, and P. megacephalus, from campos rupestres ecosystem. We sampled one mitochondrial, five nuclear loci, and 18 morphometric variables. Using species-delimitation methods with genetic and morphometric data, we recovered five divergent lineages within P. rohdei and no cryptic lineages were recovered for P. megacephalus. Morphometric data show differentiation only for one of the candidate species revealed by the delimitation approaches, suggesting that individuals from Doce River basin constitute a putative species for formal taxonomic description. The time-calibrated mtDNA tree shows that P. rohdei complex lineages began to diverge in late Miocene. However, dates from the multilocus species tree are more recent, occurring in Pleistocene, and suggesting their persistence in refuges of forest and sky islands within the Atlantic Forest biome. The existence of cryptic taxa within P. rohdei is, therefore, relevant for planning conservation strategies for this species complex in the Atlantic Forest biodiversity hotspot. Univ Fed Minas Gerais, Inst Ciencias Biol, Programa Posgrad Genet, Ave Antonio Carlos,6627, BR-31270010 Belo Horizonte, MG, Brazil Univ Fed Minas Gerais, Inst Ciencias Biol, Programa Posgrad Zool, Ave Antonio Carlos,6627, BR-31270010 Belo Horizonte, MG, Brazil Univ Brasilia, Dept Ecol, Lab Ecossistemas, Campus Univ Darcy Ribeiro, BR-70910900 Brasilia, DF, Brazil Univ Estadual Paulista, Dept Zool, Ave 24 A,1515, BR-13506900 Rio Claro, SP, Brazil Univ Estadual Paulista, Inst Biociencias, Ctr Aquicultura CAUNESP, Ave 24 A,1515, BR-13506900 Rio Claro, SP, Brazil Univ Fed Rio de Janeiro, Museu Nacl, Dept Vertebrados, Herpetol, BR-20940040 Rio De Janeiro, RJ, Brazil Univ Estadual Paulista, Dept Zool, Ave 24 A,1515, BR-13506900 Rio Claro, SP, Brazil Univ Estadual Paulista, Inst Biociencias, Ctr Aquicultura CAUNESP, Ave 24 A,1515, BR-13506900 Rio Claro, SP, Brazil FAPEMIG: APQ-01998-17 FAPEMIG: APQ-01120-16 CAPES: 440665/2015-9 FAPESP: 2012/10000-5 FAPESP: 2012/25370-2 FAPESP: 2013/50741-7 FAPESP: 2013/20423-3

Nyctimantis galeata (Pombal, Menezes, Fontes, Nunes, Rocha & Van Sluys) is a casque-headed frog member of the Lophyohylini tribe (Blotto et al. 2020), narrow-endemic to the municipality of Morro do Chapéu, Bahia state, Brazil (Pombal et al. 2012). This species was initially described as sister of Corythomantis greeningi Boulenger based on shared osteological autapomorphies within Hylidae (see Faivovich et al. 2005; Pombal et al. 2012). However, in the most recent molecular phylogenetic analysis of Lophyohylini, Blotto et al. (2020) redefined the genus Nyctimantis to accommodate species from the former genus Aparasphenodon, Argenteohyla and “Corythomantis galeata” (as initially described; Pombal et al. 2012). Currently, the genus Nyctimantis comprises seven species: N. arapapa (Pimenta, Napoli & Haddad), N. bokermanni (Pombal), N. brunoi (Miranda-Ribeiro), N. galeata, N. pomba (Assis, Santana, Silva, Quintela & Feio), N. rugiceps Boulenger, and N. siemersi (Mertens). Among these, N. arapapa (Lourenço-de-Moraes et al. 2013), N. brunoi (Wogel et al. 2006) and N. siemersi (Céspedez 2000; Cajade et al. 2010) have their tadpoles described, while oral cavity anatomy was only reported for those of N. brunoi and N. siemersi (Wogel et al. 2006; Cajade et al. 2010). Considering that larval morphology generally provides reliable information for anuran systematic and taxonomic studies (Wassersug 1980; McDiarmid & Altig 1999; Haas 2003), a formal comparison of these larvae might highlight additional non-molecular evidence that support them as congeners (Blotto et al. 2020). Herein, we describe the external morphology and oral cavity anatomy of tadpoles of N. galeata and compare it with those of related species.

Made available in DSpace on 2020-12-12T02:03:48Z (GMT). No. of bitstreams: 0 Previous issue date: 2020-07-01 The Brazilian Atlantic Forest harbors high levels of anuran diversity and endemism, including several taxa restricted to small geographic ranges. Here, we provide a multilocus phylogeny for Paratelmatobiinae, a leptodactylid subfamily composed of small-ranged species distributed in the Brazilian Atlantic Forest and in the campo rupestre ecosystem. We performed Bayesian inference and maximum likelihood analyses using three mitochondrial and five nuclear markers, and a matrix comprising a broad taxonomic sampling. We then delimitated independently evolving lineages within the group. We recovered Paratelmatobiinae and each of its four genera as monophyletic and robustly supported. Five putatively new species included in our analyses were unambiguously supported in the phylogenetic trees and delimitation analyses. We also recovered other deeply divergent and geographically structured lineages within the four genera of Paratelmatobiinae. Our estimation of divergence times indicates that diversification in the subfamily began in the Eocene and continued until the Pleistocene. We discuss possible scenarios of diversification for the four genera of Paratelmatobiinae, and outline the implications of our findings for taxonomy and conservation. Programa de Pós-Graduação em Zoologia Instituto de Biociências Universidade Estadual Paulista (UNESP), Avenida 24 A, 1515 Departamento de Biodiversidade e Centro de Aquicultura (CAUNESP) Instituto de Biociências Universidade Estadual Paulista (UNESP), Avenida 24 A, 1515 Programa de Pós-Graduação em Zoologia Instituto de Ciências Biológicas Universidade Federal de Minas Gerais Avenida Antônio Carlos, 6627 Departamento de Ciências Naturais (DCNAT) Campus Dom Bosco Universidade Federal de São João del-Rei (UFSJ), Praça Dom Helvécio, 74 Museu de Zoologia Universidade de São Paulo, Avenida Nazaré, 481 Departamento de Ciências Naturais Centro de Ciências Exatas e Naturais Fundação Universidade Regional de Blumenau Department of Biology City College of New York, 160 Convent Avenue Biology Program Graduate Center City University of New York, 365 Fifth Avenue Programa de Pós-Graduação em Zoologia Instituto de Biociências Universidade Estadual Paulista (UNESP), Avenida 24 A, 1515 Departamento de Biodiversidade e Centro de Aquicultura (CAUNESP) Instituto de Biociências Universidade Estadual Paulista (UNESP), Avenida 24 A, 1515

Clinal variation and cryptic species complexes are challenges to taxa targeted conservation. These issues are particularly problematic for the estimation of species geographic distributions, generally resulting in erroneous estimations due to unrecognized species. However, integrative taxonomy could help to solve these problems, revealing geographic and genealogical limits of species when they are not clear through morphological comparisons solely. Here we evaluated the efficiency of integrative taxonomy in identifying a new population of leaf frog from a species complex. This example was chosen because correct species identification can have an impact in the conservation assessment of a threatened species. The new population belongs to Pithecopus centralis, extending its distribution by approximately 400 km to the east of its known distribution. This identification was confirmed by the consensus between genetic, morphometric and bioacoustic evidence, even with some level of uncertainty when using each piece of evidence. We reassessed the conservation status of the species following recommendations from the International Union for Conservation of Nature (IUCN), after having checked and cleaned the data used for the species categorization in the Brazilian Red List of Threatened Fauna. In spite of the distribution extension reported here, our results demonstrate that the status of the species should be revised in the national Red List, reinforcing the importance of a rigorous taxonomic approach to improve Red Lists assessments.

Made available in DSpace on 2021-06-25T10:56:14Z (GMT). No. of bitstreams: 0 Previous issue date: 2021-07-01 We evaluated the role of Quaternary climatic fluctuations on the demographic history and population structure of amphibian species endemic to the ‘campo rupestre’ in the Neotropics, evaluating their distributional shifts, demographic changes, and lineage formation from the end of Pleistocene to present. We chose two anurans endemic to the high-elevation ‘campo rupestre’ in the Espinhaço Range (ER) in northeastern and southeastern Brazil (Bokermannohyla alvarengai and Bokermannohyla oxente), as models to test the role of Quaternary climatic fluctuations over their distribution range in this region. We collected tissue samples throughout their distribution range and used statistical phylogeography to examine processes of divergence and population demography. We generated spatial–temporal reconstructions using Bayesian inference in a coalescent framework in combination with hind-cast projections of species distribution models (SDMs). We also used the results and literature information to test alternative diversification scenarios via approximate Bayesian computation (ABC). Our results show that Quaternary climatic fluctuations influenced the geographic ranges of both species showing population expansion during the last glacial maximum (LGM) and range contraction during interglacial periods, as inferred from selected ABC models and from past projections of SDMs. We recovered Pleistocene diversification for both species occuring in distinctly unique periods for each taxon. An older and range-restricted lineage was recovered in a geographically isolated geological massif, deserving conservation and further taxonomic study. The diversification and distribution of these amphibian species endemic to the Neotropical ‘campo rupestre’ were influenced by Quaternary climatic fluctuations. The expansion of cold adapted species restricted to higher elevations during glacial periods and their concomitant retraction during interglacial periods may have been crucial for producing patterns of species richness and endemism along elevation gradients in tropical and subtropical domains. Such processes may influence the evolution of the biota distributed in heterogeneous landscapes with varied topography. Programa de Pós-graduação em Ecologia e Conservação da Biodiversidade Universidade Estadual de Santa Cruz Department of Herpetology American Museum of Natural History Herpetology Section Zoologisches Forschungsmuseum Alexander Koenig Instituto Federal Goiano – IF Goiano Departamento de Biologia Departamento de Ciências Naturais Campus Dom Bosco Universidade Federal de São João del-Rei Laboratório Sagarana Instituto de Ciências Biológicas e da Saúde Universidade Federal de Viçosa – UFV Department of Earth and Environmental Sciences Rutgers University - Newark Universidade Estadual Paulista (UNESP) Departamento de Biodiversidade Instituto de Biociências and Centro de Aquicultura (CAUNESP), Campus Rio Claro Universidade Estadual Paulista (UNESP) Departamento de Biodiversidade Instituto de Biociências and Centro de Aquicultura (CAUNESP), Campus Rio Claro

Made available in DSpace on 2018-12-11T17:23:37Z (GMT). No. of bitstreams: 0 Previous issue date: 2017-01-01 Protected areas (PAs) are essential for biodiversity conservation, but their coverage is considered inefficient for the preservation of all species. Many species are subdivided into evolutionarily significant units (ESUs) and the effectiveness of PAs in protecting them needs to be investigated. We evaluated the usefulness of the Brazilian PAs network in protecting ESUs of the critically endangered Pithecopus ayeaye through ongoing climate change. This species occurs in a threatened mountaintop ecosystem known as campos rupestres. We used multilocus DNA sequences to delimit geographic clusters, which were further validated as ESUs with a coalescent approach. Ecological niche modeling was used to estimate spatial changes in ESUs' potential distributions, and a gap analysis was carried out to evaluate the effectiveness of the Brazilian PAs network to protect P. ayeaye in the face of climate changes. We tested the niche overlap between ESUs to gain insights for potential management alternatives for the species. Pithecopus ayeaye contains at least three ESUs isolated in distinct mountain regions, and one of them is not protected by any PA. There are no climatic niche differences between the units, and only 4% of the suitable potential area of the species is protected in present and future projections. The current PAs are not effective in preserving the intraspecific diversity of P. ayeaye in its present and future range distributions. The genetic structure of P. ayeaye could represent a typical pattern in campos rupestres endemics, which should be considered for evaluating its conservation status. Programa de Pós-Graduação em Zoologia Instituto de Ciências Biológicas Universidade Federal de Minas Gerais - UFMG Belo Horizonte, Minas Gerais Brasil Programa de Desarrollo Universitario Centro Universitario de Rivera Universidad de la República - UdelaR Rivera Uruguay Laboratório de Herpetologia Departamento de Zoologia Instituto de Biociências de Rio Claro Universidade Estadual Júlio Mesquita Filho - UNESP Rio Claro, São Paulo Brasil Centro de Sensoriamento Remoto Instituto de Geociências Universidade Federal de Minas Gerais - UFMG Belo Horizonte, Minas Gerais Brasil Laboratório de Fauna e Unidades de Conservação Departamento de Engenharia Florestal Faculdade de Tecnologia Universidade de Brasília - UnB Brasília, Distrito Federal Brasil Graduação em Ciências Biológicas Instituto de Ciências Biológicas Universidade Federal de Minas Gerais - UFMG Belo Horizonte, Minas Gerais Brasil Laboratório Sagarana Instituto de Ciências Biológicas e da Saúde Universidade Federal de Viçosa - UFV Florestal, Minas Gerais Brasil

Pithecopus megacephalus is a reticulated monkey–frog species endemic to the highlands of the Espinhaco Mountain Range in southeastern Brazil, an important centre of endemism in South America. This species has a discontinuous distribution and is considered “data-deficient” by the IUCN Red List, raising concerns about its conservation. Understanding the historical dynamics and connectivity of P. megacephalus populations can provide guidelines for preservation of this species in the wild. To investigate the population dynamics of P. megacephalus, we obtained multilocus DNA data for 55 individuals from different locations along the species’ known distribution. Spatial population structure, genetic diversity and demographic parameters were evaluated using population genetic and phylogeographical tools. We also evaluated its extent of occurrence and area of occupancy to investigate extinction risk of this species. We found genetic structure along P. megacephalus’ spatial distribution in the South Espinhaco Mountain Range corresponding to three population groups: northern, central and southern. Our results could provide important data on geographic distribution and population dynamics for a Data Deficient species. Therefore, we suggest these population data, together with the species’ limited occurrence in sky island environments could be used for a more accurate classification of P. megacephalus in the IUCN list, and conservation strategies for the species should be planned accordingly.

The West Indian (Trichechus manatus) and Amazonian (T. inunguis) manatees have a sympatric occurrence at the mouth of the Amazon River. A result of this interspecific encounter is the occurrence of hybrids, which are frequently found along the coasts of Amapá state in Brazil, French Guiana and Guyana. Here we present new genetic evidence indicating the occurrence of a hybrid swarm along the Guianas Shield coastline, which is an interspecific hybrid zone that also separates T. manatus populations located east (Brazil) and west (Caribbean, Gulf of Mexico, Florida and Antilles). In addition, we suggest that this hybrid population occupies a peculiar mangrove-rich environment under strong influence of the Amazon River plume, which requires an independent management and should be considered a special conservation area.

Made available in DSpace on 2020-12-12T02:03:49Z (GMT). No. of bitstreams: 0 Previous issue date: 2020-12-01 Crossodactylodes is a poorly known genus of small-sized bromeligenous frogs, endemic to Brazil. They have a patchy distribution across the mountains of the Atlantic Forest and the campo rupestre ecosystem. To better resolve their evolutionary relationships, we performed phylogenetic analyses using a multigene DNA matrix and representative sampling within the genus. We then evaluated the evolution of phenotypical and natural history traits with the inferred phylogeny. We recovered Crossodactylodes as monophyletic, diagnosed by seven putative synapomorphies in morphological and natural history characters. Evidence supports some morphological synapomorphies as adaptations to the bromeligenous habit. We found high genetic distances among closely distributed lineages within C. bokermanni and C. izecksohni. Some of these lineages might represent undescribed cryptic species. We provide detailed accounts for each species including data on their geographic range, conservation, and natural history. All species of Crossodactylodes occur in highly threatened environments, are restricted to very small geographic ranges, and probably have limited dispersal capacity due to their small body size and dependence on bromeliads. These factors emphasize the need for habitat protection to safeguard species viability. Programa de Pós-graduação em Zoologia Instituto de Biociências Universidade Estadual Paulista (UNESP), Caixa Postal 199 Programa de Pós-graduação em Zoologia Instituto de Ciências Biológicas Departamento de Zoologia Universidade Federal de Minas Gerais Departamento de Ciências Naturais Universidade Federal de São João Del-Rei Projeto Bromeligenous Instituto Marcos Daniel Departamento de Ciências Biológicas Universidade Do Estado de Mato Grosso Departamento de Ciências Biológicas Universidade Estadual de Santa Cruz Sagarana Lab Instituto de Ciências Biológicas e da Saúde Universidade Federal de Viçosa Campus Florestal Departamento de Biologia Estrutural e Funcional Instituto de Biologia Universidade Estadual de Campinas Universidade Estadual Paulista (UNESP) Instituto de Biociências Departamento de Biodiversidade e Centro de Aquicultura (CAUNESP), Caixa Postal 199 Programa de Pós-graduação em Zoologia Instituto de Biociências Universidade Estadual Paulista (UNESP), Caixa Postal 199 Universidade Estadual Paulista (UNESP) Instituto de Biociências Departamento de Biodiversidade e Centro de Aquicultura (CAUNESP), Caixa Postal 199

Bokermannohyla (tribe Cophomantini) is an endemic genus of Brazilian treefrogs containing 32 species arranged into four phenetic groups. The B. pseudopseudis group includes nine species, which are typically found in rupestrian ecosystems of disjunct Brazilian mountain ranges. Tadpoles have been shown to be important sources of information for the taxonomy of this genus, although careful analyses of character variation and descriptions of internal anatomy remain scarce. Since the first description of B. pseudopseudis tadpoles was based on a mixed series containing specimens of B. pseudopseudis and B. sapiranga, we describe the larvae of these two species from their type localities, including their external morphology, oral cavity, and skeletal system. Tadpoles of B. pseudopseudis are easily distinguished from those of B. sapiranga by aspects of their coloration and characters of their oral disc (more labial tooth rows, more numerous submarginal papillae and flaps with labial teeth). These morphological differences may be diagnostic for these two similar species, whereas their chondrocranium, hyobranchial apparatus, and oral cavity are alike, suggesting less variability in these character systems between closely related species. Finally, we provide the first comparison of data on the skeletal system of Bokermannohyla and other Cophomantini, highlighting characters of potential relevance to the systematics of the tribe.

The evolutionary processes underlying the high diversity and endemism in the Cerrado, the most extensive Neotropical savanna, remain unclear, including the factors promoting the presence and evolution of savanna enclaves in the Amazon forest. In this study, we investigated the effects of past climate changes on genetic diversity, dynamics of species range and the historical connections between the savanna enclaves and Cerrado core for Qualea grandiflora, a tree species widely distributed in the biome. Totally, 40 populations distributed in the Cerrado core and Amazon savannas were analyzed using chloroplast and nuclear DNA sequences. We used phylogeographic, coalescent and ecological niche modeling approaches. Genetic data revealed a phylogeographic structure shaped by Pleistocene climatic oscillations. An eastern-western split in the Cerrado core was observed. The central portion of the Cerrado core harbored most of the sampled diversity for cpDNA. Ecological niche models predicted the presence of a large historical refuge in this region and multiple small refuges in peripheral areas. Relaxed Random Walk (RRW) models indicated the ancestral population in the north-western border of the central portion of the Cerrado core and cyclical dynamics of colonization related to Pleistocene climatic oscillations. Central and western ancient connections between Cerrado core and Amazonian savannas were observed. No evidence of connections among the Amazonian savannas was detected. Our study highlights the importance of Pleistocene climatic oscillations for structuring the genetic diversity of Q. grandiflora and complex evolutionary history of ecotonal areas in the Cerrado. Our results do not support the recent replacement of a large area in the Amazon forest by savanna vegetation. The Amazonian savannas appear to be fragmented and isolated from each other, evolving independently a long ago.

Protected areas (PAs) are essential for biodiversity conservation, but their coverage is considered inefficient for the preservation of all species. Many species are subdivided into evolutionarily significant units (ESUs) and the effectiveness of PAs in protecting them needs to be investigated. We evaluated the usefulness of the Brazilian PAs network in protecting ESUs of the critically endangered Pithecopus ayeaye through ongoing climate change. This species occurs in a threatened mountaintop ecosystem known as campos rupestres. We used multilocus DNA sequences to delimit geographic clusters, which were further validated as ESUs with a coalescent approach. Ecological niche modeling was used to estimate spatial changes in ESUs’ potential distributions, and a gap analysis was carried out to evaluate the effectiveness of the Brazilian PAs network to protect P. ayeaye in the face of climate changes. We tested the niche overlap between ESUs to gain insights for potential management alternatives for the species. Pithecopus ayeaye contains at least three ESUs isolated in distinct mountain regions, and one of them is not protected by any PA. There are no climatic niche differences between the units, and only 4% of the suitable potential area of the species is protected in present and future projections. The current PAs are not effective in preserving the intraspecific diversity of P. ayeaye in its present and future range distributions. The genetic structure of P. ayeaye could represent a typical pattern in campos rupestres endemics, which should be considered for evaluating its conservation status.

Male-biased sexual size dimorphism (SSD), or a lack of SSD, in amphibians may be related to the territoriality. Male-biased SSD is quite abundant among the species of the Neotropical hylid genus Bokermannohyla. However, direct observations of territorial behaviour such as combat, suggestive of the presence of a male-biased SSD, are rare. We evaluated SSD in B. martinsi and provided a field record of a male–male combat event. We found males to have significantly wider forearm and longer tibia than females. It appears that male forearm hypertrophy is related to territoriality, but we cannot reject the possibility that it is an adaptation to breeding in lotic habitats. Finally, we emphasize the importance of direct natural history observations for understanding patterns of SSD among anurans in particular and amphibians in general.

The genus Odontophrynus Reinhardt & Lutken comprises 11 species distributed throughout South and East of South America (Frost 2016) clustered in three phenetic groups: O. americanus , O. cultripes , and O. occidentalis (Savage & Cei 1965; Cei 1987; Caramaschi & Napoli 2012). Only the O. occidentalis group is recovered as monophyletic in the most recent phylogenetic hypotheses (Pyron & Wiens 2011; Pyron 2014). In these analyses, the O. cultripes group is not recovered and the monophyly of O. americanus group was not tested, since only one species of the group was included (Pyron & Wiens 2011; Pyron 2014). Therefore, the relationships within the genus are still inconclusive due to low support values and limited taxon sampling.

We describe a new species of hylid frog, Bokermannohyla sapiranga sp. nov., from Central Brazil (15°55’49” S,47°52’59” W, 1110m asl). The species is characterized by medium size (males snout-vent length 45.6±4.7mm, N=13; fe-males 46.9±6.2, N=4) for the B. pseudopseudis group and by iris color varying from orange to reddish. The advertisementcall is similar in some aspects to calls of B. pseudopseudis, but differs mainly by structure and spectral features, but alsotemporal ones such as call length, note duration, and number of notes per call. The new species uses streams with muddybeds in gallery forests as well as rocky bed streams in the highlands of Goias State and Distrito Federal. The new speciesis tentatively included in the B. pseudopseudis group based on shared morphological features, ecology, and behavior. Mor-phometric analysis of size-independent variables indicated that B. sapiranga differs from B. pseudopseudis mostly in head length and width, interorbital distance, and diameter of the fourth toe disc.

CAPES - Coordenação de Aperfeiçoamento de Pessoal de Nível Superior A conservação da ictiofauna brasileira está enfrentando uma ameaça crescente, que destaca a importância de novas ferramentas para o enriquecimento das medidas de conservação de peixes. Assim, a Genética Forense e o desenho de marcadores moleculares surgem para melhorar as atividades de controle da pesca ilegal e a detecção de espécies em amostras mal preservadas. A escolha da espécie alvo deste estudo, Hoplias intermedius, está relacionada ao impacto ambiental das bacias onde a espécie se encontra distribuída, sua crescente importância como recurso pesqueiro e a grande complexidade taxonômica do gênero. Os objetivos do nosso estudo incluem sequenciamento e montagem do genoma mitocondrial completo, desenho de primers específicos mitocondriais para H. intermedius e sua validação como marcador molecular mitocondrial visando sua aplicação com fins forenses. O DNA foi isolado e uma biblioteca genômica foi construída com o uso de sequenciamento de nova geração. Os fragmentos foram obtidos com o kit Nextera DNA Sample Preparation e com o kit de reagentes MiSeq V3-600. A montagem do mtDNA foi feita com o software CLC Workbench v8.5.1, obtendo uma cobertura de 304,45%. A anotação do genoma mitocondrial foi realizada na plataforma MITOfish, mostrando um arranjo mitogenômico típico de vertebrados (13 genes codificadores de proteínas, dois genes de RNA ribossômicos, 22 genes transportadores de RNA e uma região de controle). O software MEGA6 foi utilizado para o alinhamento de sequências mitocondriais de H. intermedius. A detecção de regiões informativas para a espécie-alvo foi validada pelo pacote do R, SPIDER. Foram realizados também testes de sensibilidade, especificidade e eficiência utilizando amostras coletadas em campo, amostras de coleções científicas e amostras de peixarias. As reações de PCR foram padronizadas para cada combinação de primers utilizando Taq polimerase 5U/ul, 2 mM dNTPs, 2M KCl e combinações específicas de soluções tampão IB, IC, IIC e 10X IVB, (Phoneutria) e DMSO. A faixa de temperatura de anelamento variou entre de primers Hin5F-Hin3R e Hin5F-Hin4R foram altamente específicos para H. intermedius, ambos projetados na região 16S rRNA. As sequências de amplificação foram obtidas para ambos os marcadores usando sequenciamento bidirecional e posteriormente foi construída uma árvore NJ para cada marcador. Nossos resultados demonstram que os marcadores moleculares para H. intermedius têm um enorme potencial para identificar a espécies-alvo, e sua aplicação inclui taxonomia integrativa para espécies de Hoplias e identificação forense usando amostras com DNA de baixa qualidade. The conservation of Brazilian ichthyofauna is facing a growing threat, which highlights the importance of new tools for the enrichment of conservation measures. Thus, Forensic Genetics and the design of molecular markers, emerge to improve illegal fishing control activities and species detection on poorly preserved samples. The choice of the target species of this study, Hoplias intermedius (Günther, 1864), responds to the environmental impact of the basins where species is distributed, its increasing importance as a fishing resource and the great taxonomic complexity of the genus. The objectives of our study include the sequencing and assembly of the whole mitochondrial genome, design of mitochondrial species-specific primers for H. intermedius and its validation as a mitochondrial molecular marker for its applications in forensic aims. DNA was extracted and a genomic library was constructed by using new generation sequencing. Fragments were obtained with Nextera DNA Sample Preparation kit and MiSeq Reagent kit V3-600 paired-end. The mtDNA assembly was made with CLC Workbench software v8.5.1 obtaining coverage of 304.45% and its annotation was performed on the MITOfish platform, showing a typical mitogenome arrangement of vertebrates (13 protein coding genes, two ribosomal RNA genes, 22 RNA transporter genes and one control region). MEGA6 software was used for the alignment of mitochondrial sequences of H. intermedius and other mtDNA with 99% of identity. The detection of informative sequence regions for the target species was validated by SPIDER R-package. Sensitivity, specificity and efficiency tests were performed by using samples collected in the field, from scientific collections and from fishmongers. PCR reactions were standardized for each primer combination using 5U/ul Taq Polymerase, 2mM dNTPs, and specific combinations of buffer solutions IB, IC, IIC and 10X IVB, 2M KCl (Phoneutria) and DMSO. The annealing temperature range varied among 59°, 62° and 64° C and the number of cycles between 30 and 35. Results showed that the mtDNA of H. intermedius is a circular DNA molecule of 16629 bp, with 43.97% GC content. The ATG start codon was found in 13 non-coding protein genes encoded in the (+) strand, and as expected the ND6 gene encoded in the (-) strand. Primer pairs Hin5F-Hin3R and Hin5F-Hin4R were highly specific for H. intermedius, both design for the 16S rRNA region. Amplicon sequences were obtained by using bidirectional sequencing, and subsequently constructed an NJ tree for each marker. Our results demonstrate that molecular markers for H. intermedius pursue a huge potential to identify the target species, and its application includes integrative taxonomy for Hoplias species and wildlife forensic identification of poor DNA quality samples.