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The diversification processes underlying why Amazonia hosts the most species-rich vertebrate fauna on earth remain poorly understood. We studied the spatio-temporal diversification of a tree frog clade distributed throughout Amazonia (Anura: Hylidae: Osteocephalus, Tepuihyla, and Dryaderces) and tested the hypothesis that Miocene mega wetlands located in western and central Amazonia impacted connectivity among major biogeographic areas during extensive periods. We assessed the group’s diversity through DNA-based (16S rRNA) species delimitation to identify Operational Taxonomic Units (OTUs) from 557 individuals. We then selected one terminal for each OTU (n = 50) and assembled a mitogenomic matrix (~14,100 bp; complete for 17 terminals) to reconstruct a Bayesian, time-calibrated phylogeny encompassing nearly all described species. Ancestral area reconstruction indicates that each genus was restricted to one of the major Amazonian biogeographic areas (western Amazonia, Guiana Shield and Brazilian Shield, respectively) between ~10 and 20 Mya, suggesting that they diverged and diversified in isolation during this period around the Pebas mega wetland. After 10 Mya and the transition to the modern configuration of the Amazon River watershed, most speciation within each genus continued to occur within each area. In Osteocephalus, only three species expanded widely across Amazonia (Osteocephalus was probably driven by climatic stability, habitat diversity and the acquisition of new reproductive modes along the Andean foothills and western Amazonia. Our findings add evidence to the importance of major hydrological changes during the Miocene on biotic diversification in Amazonia.

Major historical landscape changes have left significant signatures on species diversification. However, how these changes have affected the build-up and maintenance of Amazonia’s megadiversity continues to be debated. Here, we addressed this issue by focusing on the evolutionary history of a pan-Amazonian toad genus that has diversified throughout the Neogene (Amazophrynella). Based on a comprehensive spatial and taxonomic sampling (286 samples, all nominal species), we delimited operational taxonomic units (OTUs) from mitochondrial DNA sequences. We delimited 35 OTUs, among which 13 correspond to nominal species, suggesting a vast underestimation of species richness. Next, we inferred time-calibrated phylogenetic relationships among OTUs based on complete mitogenomic data, which confirmed an ancient divergence between two major clades distributed in eastern and western Amazonia, respectively. Ancestral area reconstruction analyses suggest that the Andean foothills and the Brazilian Shield region represent the ancient core areas for their diversification. These two clades, probably isolated from one other by lacustrine ecosystems in western Amazonia during the Miocene, display a pattern of northward and eastward dispersals throughout the Miocene‒Pliocene. Given the ecological association of Amazophrynella with non-flooded forests, our results reinforce the perception that ancient Amazonian landscape changes had a major impact on the diversification of terrestrial vertebrates.

Supplemental material for this article can be accessed here: https://doi.org/10.1080/14772000.2022.2130464; International audience; Determining the relative importance of dispersal and vicariance events across neotropical regions is a major goal in biogeography. These events are thought to be related to important landscape changes, notably the transition of Amazonia toward its modern hydrological configuration ca. 10 million years ago. We investigated the spatio-temporal context of the diversification of one of the major lineages of Pristimantis, a widespread and large genus of direct-developing Neotropical frogs. We gathered a spatially and taxonomically extensive sampling of mitochondrial DNA sequences from 754 Pristimantis gr. conspicillatus specimens, which led to delimiting 75 Operational Taxonomic Units (OTUs). Complete mitogenomes of 35 of these OTUs were assembled and collated with two nuDNA loci to reconstruct a time-calibrated phylogeny. We identified five major clades that diverged around the Oligocene-Miocene transition and that are largely restricted to distinct Neotropical regions i.e. Western Amazonia (P. conspicillatus clade), the Brazilian Shield (P. fenestratus clade), the Atlantic Forest (P. ramagii clade), the Guiana Shield (P. vilarsi clade) and the northern Andes (P. nicefori clade). The majority of the diversification events within these clades occurred in-situ from the early Miocene onward. Yet, a few ancient dispersal/vicariance events are inferred to have occurred among trans-Andean forests, the Atlantic Forest, the Brazilian and the Guiana Shields, but almost none in the last 10 Ma. The radical landscape transformations during the Miocene caused by the Andean orogeny and hydrological barriers such as the Pebas System and the subsequent transcontinental configuration of the Amazon drainage is a likely explanation for the isolation of the different clades within the P. gr. conspicillatus.

With 566 species, the neotropical genus Pristimantis is the most speciose vertebrate genus. As a result of its striking diversity, taxonomic reviews remain a challenge. Herein, we present an updated phylogeny of the Pristimantis lacrimosus group and describe a new species from Llanganates and Sangay National Parks. We also report, for the first time, the phylogenetic position of Pristimantis degener, P. eugeniae, P. katoptroides, and P. petersi. Based on our phylogeny, we add two species to the Pristimantis lacrimosus group. Through the integration of molecular and bioacoustic evidence, we describe a new species which was hidden under “Pristimantis petersi”. Pristimantis petersioidessp. nov. is most closely related to Pristimantis petersi and an undescribed species from Peru. It can be distinguished from P. petersi by its advertisement call and large genetic differences (uncorrected p-genetic distances 7.9% to 8.4% for gene 16S). Moreover, the new species and P. petersi are not sister species. We suggest assigning the new species to the Endangered Red List category because it has a small distribution range with deforestation as result of agriculture and other anthropogenic influences.

We conducted a long-term inventory of the herpetofauna of Wildsumaco Wildlife Sanctuary in the eastern part of the Napo Province in Ecuador. This private preserve is about 500 ha in size and is located on the southern slopes of Volcán Sumaco. The preserve contains primary forest, secondary forest, and pasture habitats. Based mostly on nocturnal transect sampling we documented 39 species of amphibians including one species of salamander, two species of caecilians, and 36 species of frogs. Rain frogs (Pristimantis Jiminez de la Espada, 1871) were diverse with 14 species documented. A diverse species assemblage of 45 reptile species was also documented on the preserve. Six amphibian species found were listed by the IUCN Red List. At least three species exhibited substantial geographic range extensions and seven species showed elevational range extensions. We discovered several undescribed species including one salamander, three frogs, one lizard, and one snake.

The outstanding biodiversity of the Guiana Shield has raised many questions about its origins and evolution. Frogs of the genera Adelastes, Otophryne and Synapturanus form an ancient lineage distributed mostly across this region. These genera display strikingly disparate morphologies and life-history traits. Notably, Synapturanus is conspicuously adapted to fossoriality and is the only genus within this group to have dispersed further into Amazonia. Moreover, morphological differences among Synapturanus species suggest different degrees of fossoriality that might be linked to their biogeographical history. Through integrative analysis of genetic, morphometric and acoustic data, we delimited 25 species in this clade, representing a fourfold increase. We found that the entire clade started to diversify ~55 Mya and Synapturanus ~30 Mya. Members of this genus probably dispersed three times out of the Guiana Shield both before and after the Pebas system, a wetland ecosystem occupying most of Western Amazonia during the Miocene. Using a three-dimensional osteological dataset, we characterized a high morphological disparity across the three genera. Within Synapturanus, we further characterized distinct phenotypes that emerged concomitantly with dispersals during the Miocene and possibly represent adaptations to different habitats, such as soils with different physical properties., Agence Nationale de la Recherche (CEBA, ref. ANR-10-LABX-25-01), Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq #140716/2016-5; 305535/2017-0; PQ 302501/2019-3; PQ 303070/2017-0

The adaptive role of amphibian oocyte melanic pigmentation and its molecular control are still elusive. Here we present evidence of a polymorphism in egg pigmentation in the emerald glass frog Espadarana prosoblepon. In Ecuadorian natural populations of this species, females can lay dark brown or pale eggs that develop into normal pigmented tadpoles and adults. This trait is a sex-limited phenotype which is inherited like a recessive allele that we called pale eggs like (pel). The pel phenotype is exclusive of oocyte cortical melanic pigmentation, which is reduced in comparison to wild type (wt) dark pigmented oocytes. Consequently, pel early embryos are paler in appearance, with reduced melanic pigmentation distributed to early blastomeres and embryonic ectoderm. However, these embryos form normal melanocyte derived pigmentation. Finally, we discuss the origin of this polymorphism and propose the use of E. prosoblepon as a model to study the adaptive role of egg pigmentation.

Field biology is an area of research that involves working directly with living organisms in situ through a practice known as “fieldwork.” Conducting fieldwork often requires complex logistical planning within multiregional or multinational teams, interacting with local communities at field sites, and collaborative research led by one or a few of the core team members. However, existing power imbalances stemming from geopolitical history, discrimination, and professional position, among other factors, perpetuate inequities when conducting these research endeavors. After reflecting on our own research programs, we propose four general principles to guide equitable, inclusive, ethical, and safe practices in field biology: be collaborative, be respectful, be legal, and be safe. Although many biologists already structure their field programs around these principles or similar values, executing equitable research practices can prove challenging and requires careful consideration, especially by those in positions with relatively greater privilege. Based on experiences and input from a diverse group of global collaborators, we provide suggestions for action-oriented approaches to make field biology more equitable, with particular attention to how those with greater privilege can contribute. While we acknowledge that not all suggestions will be applicable to every institution or program, we hope that they will generate discussions and provide a baseline for training in proactive, equitable fieldwork practices.

Molecular phylogenies have yielded strong support for many parts of the amphibian Tree of Life, but poor support for the resolution of deeper nodes, including relationships among families and orders. To clarify these relationships, we provide a phylogenomic perspective on amphibian relationships by developing a taxon-specific Anchored Hybrid Enrichment protocol targeting hundreds of conserved exons which are effective across the class. After obtaining data from 220 loci for 286 species (representing 94% of the families and 44% of the genera), we estimate a phylogeny for extant amphibians and identify gene tree–species tree conflict across the deepest branches of the amphibian phylogeny. We perform locus-by-locus genealogical interrogation of alternative topological hypotheses for amphibian monophyly, focusing on interordinal relationships. We find that phylogenetic signal deep in the amphibian phylogeny varies greatly across loci in a manner that is consistent with incomplete lineage sorting in the ancestral lineage of extant amphibians. Our results overwhelmingly support amphibian monophyly and a sister relationship between frogs and salamanders, consistent with the Batrachia hypothesis. Species tree analyses converge on a small set of topological hypotheses for the relationships among extant amphibian families. These results clarify several contentious portions of the amphibian Tree of Life, which in conjunction with a set of vetted fossil calibrations, support a surprisingly younger timescale for crown and ordinal amphibian diversification than previously reported. More broadly, our study provides insight into the sources, magnitudes, and heterogeneity of support across loci in phylogenomic data sets.[AIC; Amphibia; Batrachia; Phylogeny; gene tree–species tree discordance; genomics; information theory.]

The combination of genetic and phenotypic characters for species delimitation has allowed the discovery of many undescribed species of Neotropical amphibians. In this study, we used DNA sequences (genes 12S, 16S, ND1 and COI) and morphologic, bioacoustic and environmental characters of the Boana semilineata group to evaluate their phylogenetic relationships and assess their species limits. In addition, we included DNA sequences of several species of Boana to explore cryptic diversity in other groups. We found three Confirmed Candidate Species (CCS) within the B. semilineata group. Holotype examination of Hyla appendiculata shows that it is a valid species that corresponds to one of the CCS, which is here transferred to Boana. We describe the two remaining CCS. Our phylogeny highlights a number of secondary but meaningful observations that deserve further investigation: (1) populations of B. pellucens from northern Ecuador are more closely related to B. rufitela from Panama than to other Ecuadorian populations of B. pellucens; (2) we report, for the first time, the phylogenetic relationships of B. rubracyla showing that it is closely related to B. rufitela and B. pellucens; and (3) B. cinerascens and B. punctata form two species complexes consisting of several unnamed highly divergent lineages. Each of these lineages likely represents an undescribed species.

The Osteocephalus buckleyi species group is widely distributed in primary and secondary forests of the Amazon Basin and Guiana Region. Based on integrative analysis, including morphological and genetic data, we estimate the phylogenetic relationships and species boundaries among populations of the Osteocephalus buckleyi group from the Ecuadorian Amazon, focusing on the O. verruciger-O. cannatellai species complex. Our results uncovered the existence of one confirmed candidate species from Sangay National Park and one unconfirmed candidate species. Here, we describe the new species which is morphologically and ecologically distinct from other Osteocephalus species. The new species is unusual because it shows quite distinct morphology, but low genetic distances compared to its closest relatives. Zoobank.org registration LSID: urn:zoobank.org:pub:01F86A33-2D07-4D9E-A8A2-FF35622E3DB9

Species richness in the genus Pristimantis is underestimated due to the existence of morphologically cryptic species. This is worsened by the low sampling effort and the lack of studies using genetic markers. Here, we use molecular and morphological data to determine the phylogenetic relationships of a clade of Pristimantis distributed throughout montane tropical forests in the eastern Andes, from central Ecuador to northern Perú. We name this clade the Pristimantis trachyblepharis species group. Our results show that it comprises nine species, of which four are formally described and five are new. Four of these undescribed species are formally described here. The group is composed of miniaturized species, such as Pristimantis nanus sp. nov., currently the smallest known species of the genus and the smallest vertebrate in Ecuador. As a first approach to understanding the evolutionary origin and implications of body-size reduction in Pristimantis, we here study the phylogenetic signal and evolutionary trends of body size within the genus. We also provide the first record of P. aquilonaris in Ecuador and we show, for the first time, the phylogenetic position of P. albujai, P. aquilonaris, P. minimus and P. trachyblepharis, which are also members of the P. trachyblepharis species group.

Microbiome-pathogen interactions are increasingly recognised as an important element of host immunity. While these host-level interactions will have consequences for community disease dynamics, the factors which influence host microbiomes at larger scales are poorly understood. We here describe landscape scale pathogen-microbiome associations within the context of post-epizootic amphibian chytridiomycosis, a disease caused by the panzootic chytrid fungus Batrachochytrium dendrobatidis. We undertook a survey of Neotropical amphibians across altitudinal gradients in Ecuador ~30 years following the observed amphibian declines and collected skin swab-samples which were metabarcoded using both fungal (ITS-2) and bacterial (r16S) amplicons. Our data reveals marked variation in patterns of both B. dendrobatidis infection and microbiome structure that are associated with host life history. Stream breeding amphibians were most likely to be infected with B. dendrobatidis. This increased probability of infection was further associated with increased abundance and diversity of non-Batrachochytrium chytrid fungi in the skin and environmental microbiome. We also show that increased alpha diversity and the relative abundance of fungi is lower in the skin microbiome of adult stream amphibians compared to adult pond breeding amphibians, an association not seen for bacteria. Finally, stream tadpoles exhibit lower proportions of predicted protective microbial taxa than pond tadpoles, suggestive of reduced biotic resistance. Our analyses show that host breeding ecology strongly shapes pathogen-microbiome associations at a landscape scale, a trait that may influence resilience in the face of emerging infectious diseases.

Made available in DSpace on 2021-06-25T12:41:13Z (GMT). No. of bitstreams: 0 Previous issue date: 2021-01-20 'Investissement d'Avenir' grant Brazilian Fundacao de Amparo a Pesquisa do Estado do Amazonas Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) SENESCYT (Arca de Noe Initiative) Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) Centro de Estudos de Insetos Sociais (CEIS) The outstanding species richness of Amazonia has fascinated biologists for centuries. However, the records of actual numbers and distribution of species forming its ecosystems are so incomplete that the understanding of the historical causes and regional determinants of this diversity remain speculative. Anuran clades have repeatedly been documented to harbour many unnamed species in this region, notably the Boana albopunctata species group. Considering the documented distribution and the ecology of the species of that group, we hypothesized that it diversified via successive trans-riverine dispersals during the late Miocene and Pliocene, after the formation of the modern Amazon watershed. To test this hypothesis, we gathered an extensive dataset of 16S rDNA sequences sampled throughout Amazonia and a mitogenomic dataset representative of the diversity of the Glade to (1) re-evaluate species boundaries and distributions, and (2) infer the spatio-temporal history of diversification within Amazonia. We delimited 14 Operational Taxonomic Units (OTUs) in an Amazonian Glade, i.e., 75% higher than currently recognized (14 OTUs for eight described species). Combining molecular data with morphological and acoustic data, two new species, Boana courtoisae sp. nov. from the eastern Guiana Shield and Boana eucharis sp. nov. from Southern Amazonia, are described herein. These species belong to a Glade that diversified throughout Amazonia during the last 10 Ma, thus more recently than co-distributed small terrestrial anurans but concomitantly with other more vagile vertebrates. Our time-scaled phylogeny and biogeographic analyses suggest an initial east-west divergence and confirm reciprocal trans-riverine dispersals during the last 5 Ma. The geomorphological evolution of the region and species-specific dispersal ability largely explain these distinct spatio-temporal patterns across anurans. http://www.zoobank.org/zoobank.org:act:4F8ACA9F-F6F1-4605-BD6C-6D4650AACCBE http://www.zoobank.org/zoobank.org:act:51CC7B40-2D6B-4A9E-AF50-AB34D4CE1042 Univ Paul Sabatier, Lab Evolut & Diversite Biol, CNRS, UMR 5174,IRD, Batiment 4R1,118 Route Narbonne, F-31062 Toulouse 9, France Univ Fed Uberlandia, Inst Ciencias Exatas & Nat Pontal, Lab Anuros Neotrop, Ituiutaba, MG, Brazil Univ Estadual Paulista, Dept Biodiversidade, Lab Herpetol, IB, Rio Claro, SP, Brazil Univ Estadual Paulista, Ctr Aquicultura, IB, Rio Claro, SP, Brazil Pontificia Univ Catolica Ecuador, Escuela Biol, Museo Zool, Quito, Ecuador Johannes Gutenberg Univ Mainz, Inst Organism & Mol Evolut, Mainz, Germany Res Inst & Nat Museum Senckenberg, Dept Terr Zool, Frankfurt, Germany Inst Nacl de Pesquisas da Amazonia, Coordenacao Biodiversidade, Manaus, Amazonas, Brazil Univ Sao Paulo, Inst Biociencias, Dept Zool, Sao Paulo, SP, Brazil Univ Fed Amazonas, Dept Genet, Manaus, Amazonas, Brazil Inst Peruano Herpetol, Lima, Peru Univ Estadual Paulista, Dept Biodiversidade, Lab Herpetol, IB, Rio Claro, SP, Brazil Univ Estadual Paulista, Ctr Aquicultura, IB, Rio Claro, SP, Brazil 'Investissement d'Avenir' grant: ANR-10-LABX-25-01 'Investissement d'Avenir' grant: ANR-10-LABX-0041 'Investissement d'Avenir' grant: ANR-11-INBS-0001 Brazilian Fundacao de Amparo a Pesquisa do Estado do Amazonas: FAPEAM10.13039/501100004916062.00962/2018 FAPESP: 2003/10335-8 FAPESP: 2011/50146-6 FAPESP: BIOTA 2013/50297-0 FAPESP: NSF-DEB 1343578 CNPq: 446935/2014-0 CNPq: 300903/2015-4 CNPq: 305169/2019-0 FAPESP: 2017/08489-0 FAPESP: 2013/50741-7 CAPES: 88887.201356/2018-00

True toads of the genus Rhinella are among the most common and diverse group of Neotropical anurans. These toads are widely distributed throughout South America, inhabiting a great diversity of environments and ecoregions. Currently, however, the genus is defined solely on the basis of molecular characters, and it lacks a proper diagnosis. Although some phenetic species groups have traditionally been recognized within Rhinella, the monophyly of some of them have been rejected in previous phylogenetic analyses, and many species remain unassigned to these poorly defined groups. Additionally, the identity and taxonomy of several species are problematic and hinder the specific recognition and description of undescribed taxa. In this work, we first perform phylogenetic analyses of separate mitochondrial and nuclear datasets to test the possible occurrence of hybridiza-tion and/or genetic introgression in the genus. The comparative analysis of both datasets revealed unidirectional mitochondrial introgressions of an unknown parental species into R . horribilis (“ghost introgression”) and of R . dorbignyi into R . bernardoi; therefore, the mitochondrial and nuclear data-sets of these species were considered separately in subsequent analyses. We performed total-evidence phylogenetic analyses that included revised molecular (four mitochondrial and five nuclear genes) and phenotypic (90 characters) datasets for 83 nominal species of Rhinella, plus several undescribed and problematic species and multiple outgroups. Results demonstrate that Rhinella was nonmono-phyletic due to the position of R . ceratophrys, which was recovered as the sister taxon of Rhaebo nasicus with strong support. Among our outgroups, the strongly supported Anaxyrus + Incilius is the sister clade of all other species of Rhinella. Once R . ceratophrys is excluded, the genus Rhinellais monophyletic, well supported, and composed of two major clades. One of these is moderately supported and includes species of the former R . spinulosa Group (including R . gallardoi); the mono-phyletic R . granulosa, R . crucifer, and R . marina Groups; and a clade composed of the mitochondrial sequences of R . horribilis. The other major clade is strongly supported and composed of all the spe-cies from the non-monophyletic R . veraguensis and R . margaritifera Groups, the former R . acrolophaGroup, and R . sternosignata. Consistent with these results, we define eight species groups of Rhinella that are mostly diagnosed by phenotypic synapomorphies in addition to a combination of morpho-logical character states. Rhinella sternosignata is the only species that remains unassigned to any group. We also synonymize nine species, treat three former subspecies as full species, and suggest that 15 lineages represent putative undescribed species. Lastly, we discuss the apparently frequent occurrence of hybridization, deep mitochondrial divergence, and “ghost introgression”; the incomplete phenotypic evidence (including putative character systems that could be used for future phy-logenetic analyses); and the validity of the known fossil record of Rhinella as a source of calibration points for divergence dating analyses.

Made available in DSpace on 2021-06-25T12:36:12Z (GMT). No. of bitstreams: 0 Previous issue date: 2021-01-01 Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) Agencia Nacional de Promocion Cientifica y Tecnologica de Argentina The Leptodactylus melanonotus species group is widely distributed across the Neotropics and contains 17 valid species. Here we reassess the taxonomic status of several taxa of the group, focusing mainly on the Amazonian L. petersii and both its junior synonyms (L. brevipes and L. intermedius), based on morphological, acoustic, cytogenetic, and DNA sequence data. The phylogenetic analyses recovered a monophyletic L. melanonotus group. However, L. petersii in the broad sense was found to be polyphyletic by containing the nominal species and two other lineages more closely related to L. podicipinus than to L. petersii. Color patterns and calls also support the distinctiveness among these lineages. Moreover, one of the lineages (L. brevipes) has a unique karyotype in the genus (2n = 20 vs. 2n = 22 in the other species). The holotype of L. petersii is presumably lost and a previous designation of a neotype was later considered as invalid. Therefore, we designate a new neotype for L. petersii and redescribe the species from the upper Negro River drainage. Also, we revalidate and characterize L. brevipes and L. intermedius from their type localities. Our study contributes to the systematics, cytogenetic and phenotypic variation, and distribution patterns of leptodactylid frogs in South America. (c) 2020 Elsevier GmbH. All rights reserved. Univ Estadual Paulista, Lab Citogenet Anim, Dept Biol Geral & Aplicada, Inst Biociencias, Rio Claro, SP, Brazil Univ Estadual Paulista, Lab Herpetol, Dept Biodiversidade, Rio Claro, SP, Brazil Univ Estadual Paulista, Inst Biociencias, Ctr Aquicultura, Rio Claro, SP, Brazil Pontificia Univ Catolica Ecuador, Museo Zool, Escuela Ciencias Biol, Quito, Ecuador Univ Fed Mato Grosso, Fac Med Vet, Cuiaba, MT, Brazil Univ Nacl Misiones, Inst Biol Subtrop, Fac Ciencias Exactas Quim & Nat, Lab Genet Evolut, Posadas, Argentina Univ Fed Mato Grosso, Inst Biociencias, Ctr Referencia Biodiversidade Reg, Lab Herpetol, Cuiaba, MT, Brazil Univ Fed Uberlandia, Lab Anuros Neotropicais, Inst Ciencias Exatas & Nat Pontal, Ituiutaba, MG, Brazil Univ Estadual Paulista, Lab Citogenet Anim, Dept Biol Geral & Aplicada, Inst Biociencias, Rio Claro, SP, Brazil Univ Estadual Paulista, Lab Herpetol, Dept Biodiversidade, Rio Claro, SP, Brazil Univ Estadual Paulista, Inst Biociencias, Ctr Aquicultura, Rio Claro, SP, Brazil FAPESP: 2008/50928-1 FAPESP: 2012/15763-7 FAPESP: 2013/50741-7 FAPESP: 2017/08489-0 FAPESP: 2017/26162-8 CNPq: 446935/2014-0 CNPq: 302518/2013-4 CNPq: 305169/2019-0 CNPq: 306623/2018-8 CNPq: 3123038/2018-1 Agencia Nacional de Promocion Cientifica y Tecnologica de Argentina: PICTs 2015/2381 Agencia Nacional de Promocion Cientifica y Tecnologica de Argentina: 2016/4066 Agencia Nacional de Promocion Cientifica y Tecnologica de Argentina: 2018/3349

We describe through integrative taxonomy a new Amazonian species of leaf-litter toad of the Rhinella margaritifera species group. The new species inhabits open lowland forest in southwest Amazonia in Brazil, Peru, and Bolivia. It is closely related to a Bolivian species tentatively identified as Rhinella cf. paraguayensis. Both the new species and R. paraguayensis share an uncommon breeding strategy among their Amazonian congeners: each breeds in moderate to large rivers instead of small streams or ponds formed by rainwater. The new species is easily differentiated from other members of the R. margaritifera species group by having a strongly developed bony protrusion at the angle of the jaw, a snout–vent length of 63.4–84.7 mm in females and 56.3–72.3 mm in males, well-developed supratympanic crests with the proximal portion shorter than the parotoid gland in lateral view, a divided distal subarticular tubercle on finger III, and multinoted calls composed of groups of 7–9 pulsed notes and a dominant frequency of 1,012–1,163 Hz. Recent studies have shown that the upper Madeira Basin harbors a megadiverse fauna of anurans, including several candidate species. This is the first member of the R. margaritifera species group to be described from this region in recent years, and at least two additional unnamed species await formal description.

Made available in DSpace on 2021-06-25T12:20:40Z (GMT). No. of bitstreams: 0 Previous issue date: 2020-08-04 Human Frontier Science Program Consejo Nacional de Investigaciones Cientificas y Tecnicas (CONICET) Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) Agencia Nacional de Promocion Cientifica y Tecnologica, Proyecto de Investigacion Cientifica y Tecnologica CONICET Universidad de Buenos Aires Universidad de Buenos Aires Ciencia y Tecnica Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) Secretaria Nacional de Educacion Superior, Ciencia, Tecnologia e Innovacion del Ecuador (SENESCYT) (Arca de Noe initiative) Direccion General Academica Pontificia Universidad Catolica del Ecuador Many vertebrates have distinctive blue-green bones and other tissues due to unusually high biliverdin concentrations-a phenomenon called chlorosis. Despite its prevalence, the biochemical basis, biology, and evolution of chlorosis are poorly understood. In this study, we show that the occurrence of high biliverdin in anurans (frogs and toads) has evolved multiple times during their evolutionary history, and relies on the same mechanism-the presence of a class of serpin family proteins that bind biliverdin. Using a diverse combination of techniques, we purified these serpins from several species of nonmodel treefrogs and developed a pipeline that allowed us to assemble their complete amino acid and nucleotide sequences. The described proteins, hereafter named biliverdin-binding serpins (BBS), have absorption spectra that mimic those of phytochromes and bacteriophytochromes. Our models showed that physiological concentration of BBS5 fine-tune the color of the animals, providing the physiological basis for crypsis in green foliage even under near-infrared light. Additionally, we found that these BBS5 are most similar to human glycoprotein alpha-1-antitrypsin, but with a remarkable functional diversification. Our results present molecular and functional evidence of recurrent evolution of chlorosis, describe a biliverdin-binding protein in vertebrates, and introduce a function for a member of the serpin superfamily, the largest and most ubiquitous group of protease inhibitors. Duke Univ, Dept Biol, Durham, NC 27708 USA Consejo Nacl Invest Cient & Tecn CONICET, Museo Argentino Ciencias Nat Bernardino Rivadavia, Div Herpetol, C1405DJR, Buenos Aires, DF, Argentina Univ Buenos Aires, Fac Ciencias Exactas & Nat, Inst Quim Fis Mat Medio Ambiente & Energia, Consejo Nacl Invest Cient & Tecn CONICET, C1428EHA, Buenos Aires, DF, Argentina Univ Sao Paulo, Fac Ciencias Farmaceut Ribeirao Preto, Dept Ciencias BioMol, Nucleo Pesquisa Prod Nat & Sintet NPPNS, BR-14040903 Ribeirao Preto, SP, Brazil Univ Nacl Misiones, Fac Ciencias Exactas, Inst Biol Subtrop CONICET UNaM, Lab Genet Evolut Claudio Juan Bidau, RA-3300 Posadas, Misiones, Argentina Univ Estadual Paulista, Dept Biodiversidade, BR-13506900 Rio Claro, SP, Brazil Univ Estadual Paulista, Ctr Aquicultura, Inst Biociencias, BR-13506900 Rio Claro, SP, Brazil Univ Cent Florida, Dept Biol Genom & Bioinformat Cluster, Orlando, FL 32816 USA Pontificia Univ CatOlica Ecuador, Escuela Biol, Museo Zool, Aptdo 17-01-2184, Quito, Ecuador Univ Buenos Aires, Fac Ciencias Exactas & Nat, Dept Quim Inorgan Analit & Quim Fis, C1428EHA, Buenos Aires, DF, Argentina Univ Buenos Aires, Fac Ciencias Exactas & Nat, Dept Biodiversidad & Biol Expt, C1428EHA, Buenos Aires, DF, Argentina Consejo Nacl Invest Cient & Tecn, Fdn Inst Leloir, C1405BWE, Buenos Aires, DF, Argentina Consejo Nacl Invest Cient & Tecn, Inst Invest Bioquim Buenos Aires, C1405BWE, Buenos Aires, DF, Argentina Univ Nacl San Martin, Inst Invest Biotecnol, RA-1650 San Martin, Buenos Aires, Argentina Univ Estadual Paulista, Dept Biodiversidade, BR-13506900 Rio Claro, SP, Brazil Univ Estadual Paulista, Ctr Aquicultura, Inst Biociencias, BR-13506900 Rio Claro, SP, Brazil Human Frontier Science Program: LT 000660/2018-L FAPESP: 2014/20915-6 FAPESP: 2017/23725-1 FAPESP: 2017/26162-8 Agencia Nacional de Promocion Cientifica y Tecnologica, Proyecto de Investigacion Cientifica y Tecnologica: 2011-1895 Agencia Nacional de Promocion Cientifica y Tecnologica, Proyecto de Investigacion Cientifica y Tecnologica: 2013-404 Agencia Nacional de Promocion Cientifica y Tecnologica, Proyecto de Investigacion Cientifica y Tecnologica: 2013-1895 Agencia Nacional de Promocion Cientifica y Tecnologica, Proyecto de Investigacion Cientifica y Tecnologica: 2014-1022 Agencia Nacional de Promocion Cientifica y Tecnologica, Proyecto de Investigacion Cientifica y Tecnologica: 2015-820 CONICET: 11220150100394CO Universidad de Buenos Aires Ciencia y Tecnica: 20020170100037BA FAPESP: 2013/50741-7 FAPESP: 2012/10000-5 FAPESP: 2014/50342-8 FAPESP: 2018/15425-0 CNPq: 306623/2018-8

Aim: We investigate the spatiotemporal context of the diversification of Allobates, a widespread genus of Amazonian frogs with high species diversity particularly in western Amazonia. We tested if that diversity originated in situ or through repeated dispersals from other Amazonian areas and if this diversification took place during or after the Pebas system, a vast lacustrine system occupying most western Amazonia between 23 and 10 million years ago (Mya). Location: Amazonia. Taxon: Allobates (Anura: Aromobatidae). Methods: We gathered a spatially and taxonomically extensive sampling of mitochondrial DNA sequences from 932 Allobates specimens to delimit Operational Taxonomic Units (OTUs). Complete mitogenomes of these OTUs were assembled to reconstruct a time-calibrated phylogeny used to infer the historical and spatial origin of the Amazonian Allobates lineages. Results: Phylogenetic analyses and ancestral area reconstruction suggested that most of the western Amazonian lineages resulted from in situ diversification and that these events occurred between the inferred origin of the genus (25 Mya) and 10 Mya, with a possible peak between 14 and 10 Mya. Dispersal among areas mostly occurred from western Amazonia towards the Brazilian and the Guiana Shields. Closely related Allobates OTUs display an allopatric pattern of distribution, matching interfluves delimited by modern Amazonian rivers. Main Conclusions: In western Amazonia, diversification of Allobates appears to have been simultaneous with the last stages of the Pebas system (14–10 Mya). Subsequently (within the last 10 Mya), modern Amazonian river courses shaped the distribution pattern of Allobates species and possibly promoted allopatric speciation., This study benefited from an “Investissement d’Avenir” grant managed by the Agence Nationale de la Recherche (CEBA, ref. ANR-10-LABX-25-01; TULIP, ref. ANR-10-LABX-0041; ANAEE-France: ANR-11-INBS-0001). AF and FPW acknowledge the French/Brazilian GUYAMAZON program action (IRD, CNRS, CTG, CIRAD and Brazilian Fundação de Amparo à Pesquisa do Estado do Amazonas-FAPEAM 062.00962/2018) and co-coordinated by A. Fouquet and F. P. Werneck. MTR thanks Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq), Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP grant numbers: 2003/10335-8, 2011/50146-6 and NSF-FAPESP Dimensions of Biodiversity Program [grant numbers: BIOTA 2013/50297-0, NSF-DEB 1343578]) and NASA. SRR acknowledges a grant from SENESCYT (Arca de Noé Initiative). FPW thanks CNPq (Productivity Fellowship), FAPEAM, Coordenação de Aperfeiçoamento de Pessoal de Nível Superior-CAPES (Visiting Professor Fellowship) and the L’Oréal-UNESCO For Women In Science Program. AFJ is supported by the Coordenação de Aperfeiçoamento de Pessoal de Nivel Superior-Brasil (CAPES Finance Code: 88882.462010/2019-01). JM is supported by the Ministry of Culture of the Czech Republic (DKRVO 2019-20023/6.VI.b, National Museum Prague, 00023272). PJRK was supported by research grants from the Fonds voor Wetenschappelijk Onderzoek (FWO12A7614N and FWO12A7617N). AJC was supported by Research Program INV-2017-51-1432 from the School of Sciences, Universidad de los Andes. RE was supported by research grants from the Deutsche Forschungsgemeinschaft (DFG ER 589/1-1 & ER 589/2-1).

Knowledge on long-term population trends in crocodilians is essential to assess the effectiveness of conservation areas and to guide sustainable management practices. We studied changes in population size of spectacled (Caiman crocodilus) and black caimans (Melanosuchus niger) over a period of 17 yr at Mateococha, a black-water lake located in Cuyabeno Wildlife Reserve, western Amazonia, Ecuador. Using standardized spotlight counts and two mark–recapture surveys, we estimated the population abundance, body size structure, and sex ratio in 1994, 2004, and 2011. The maximum number of C. crocodilus recorded in 1994 (147 individuals; 33 individuals/kilometer [ind/km] of lakeshore) declined by 2004 to 94 individuals (21.1 ind/km) and by 2011 to 63 individuals (14.2 ind/km). The number of M. niger recorded was low but constant (5–7 individuals; 1.1–1.6 ind/km) throughout the study, suggesting that factors causing the decline in C. crocodilus were not related to the M. niger population trend. Large C. crocodilus adults (total length 150–220 cm) were frequent in 1994 but became rare in 2004 and 2011. The sex ratio of captured C. crocodilus was male-biased during all periods. The causes of the population decline are unknown, but available evidence suggests that illegal hunting and habitat degradation are likely culprits despite the protected status of the Cuyabeno Reserve. Our findings question the effectiveness of protected areas to ensure long-term survival of caiman populations in Ecuador.

© 2017 The Authors., Assembly of ecological communities is important for the conservation of ecosystems, predicting perturbation impacts, and understanding the origin and loss of biodiversity. We tested how amphibian communities are assembled by neutral and niche‐based mechanisms, such as habitat filtering. Species richness, β‐diversities, and reproductive traits of amphibians were evaluated at local scale in seven habitats at different elevation and disturbance levels in Wisui Biological Station, Morona‐Santiago, Ecuador, on the foothills of the Cordillera del Kutukú; and at regional scale using 109 localities across evergreen forests of Amazonia and its Andean slopes (0–3,900 m a.s.l.). At local scale, species composition showed strong differences among habitats, explained mainly by turnover. Reproductive modes occurred differently across habitats (e.g., prevalence of direct developers at high elevation, where breeding in ground level water disappears). At regional scale, elevation was the most important factor explaining the changes in species richness, reproductive trait occurrences, and biotic dissimilarities. Species number in all groups decreased with elevation except for those with lotic tadpoles and terrestrial reproduction stages. Seasonality, annual precipitation, and relative humidity partially explained the occurrence of some reproductive traits. Biotic dissimilarities were also mostly caused by turnover rather than nestedness and were particularly high in montane and foothill sites. Within lowlands, geographic distance explained more variability than elevation. Habitat filtering was supported by the different occurrence of reproductive traits according to elevation, water availability, and breeding microhabitats at both scales, as well as other assembly mechanisms based in biotic interactions at local scale. Human‐generated land use changes in Amazonia and its Andean slopes reduce local amphibian biodiversity by alteration of primary forests and loss of their microhabitats and the interaction network that maintains their unique amphibian assemblages with different reproductive strategies., Specimens were collected under research permits 008‐09 IC‐FAU‐DNB/MA and 05‐2011‐Investigacion‐B‐DPMS from the Ministerio del Ambiente of Ecuador. This research was partly supported by projects CGL2011‐30393BOS and CGL2014‐56160‐P of the Spanish Government (PI, I. De la Riva).

Ecnomiohyla tuberculosa is an Amazonian hylid of uncertain phylogenetic position. Herein DNA se- quences of mitochondrial and nuclear genes are used to determine its phylogenetic relationships. New sequences and external morphology of Trachycephalus typhonius are also analyzed to assess the status of Ecuadorian and Peruvian populations. The phylogeny shows unequivocally that Ecnomiohyla tuberculosa is nested within the genus Tepuihyla, tribe Lophiohylini. This position was unexpected because the remain- ing species of Ecnomiohyla belong to the tribe Hylini. To solve the paraphyly of the genus Ecnomiohyla, E. tuberculosa is transferred to the genus Tepuihyla. Comparisons of DNA sequences, external morphology, and advertisement calls between populations of Ecnomiohyla tuberculosa from Ecuador and Peru indicate that the Peruvian population represents an undescribed species. The new species is described and a species Copyright Santiago R. Ron et al.. This is an open access article distributed under the terms of the Creative Commons Attribution License (CC BY 4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. account is provided for Ecnomiohyla tuberculosa. Trachycephalus typhonius is paraphyletic relative to T. cu- nauaru, T. hadroceps, and T. resinifictrix. The phylogenetic position of populations from western Ecuador indicates that they represent a species separate from T. typhonius sensu stricto. We resurrect the name Hyla quadrangulum (Trachycephalus quadrangulum comb. n.) for those populations. Amazonian populations of “T. typhonius” from Ecuador and Peru are genetically and morphologically distinct from T. typhonius sensu stricto and are conspecific with the holotype of Hyla macrotis. Therefore, we also resurrect Hyla macrotis, a decision that results in Trachycephalus macrotis comb. n. Pensoft Publishers

We describe a new species of terrestrial frog of the genus Noblella from the eastern versants of the Ecuadorian Andes in the upper Pastaza watershed. Noblella naturetrekii sp. n. differs from its Ecuadorian congeners by the presence of a differentiated tympanic membrane and a weakly defined tympanic annulus, and eyelids with rounded tubercles. In addition, the new species is characterized by its blackish–dark brown ventral coloration scattered with little white dots and the absence of papillae at the tip of the fingers and toes. We provide a detailed description of the call and osteology of the new species. Finally, we present the most complete phylogeny of the genus, which confirms that Noblella is a non-monophyletic group.

Since 1984 there have been no records of Rhaebo colomai (Hoogmoed, 1985) within the territory of Ecuador. This species was known from 2 localities in the province of Carchi, northwestern Ecuador, and the department of Nariño, southwestern Colombia, which were reported in 1979 and 2015, respectively. We report the recent sightings of R. colomai at 3 new localities in Ecuador and discuss and evaluate this species’ extinction risk and conservation status.

We describe a new species ofPristimantisfrom the montane forest of the Río Zuñag Ecological Reserve, upper basin of the Pastaza River, Ecuador.Pristimantismalliisp. n.is characterized by a snout-vent length of 11.6–21.3 mm in adult males (n= 12), 22.6–34.3 mm in adult females (n= 8), and is compared morphologically and genetically withPristimantismiktosand with other relevant species ofPristimantis. The new species is characterized by having skin on dorsum and flanks shagreen, distinctive scapular folds, snout broadly rounded in dorsal view, upper eyelid bearing one or two subconical tubercles and some rounded tubercles, dorsum and flanks light brown to brown, with irregular dark brown marks bounded by dirty cream and groin with irregular yellowish marks.

We describe two new species of frogs of the genus Pristimantis from the eastern slopes of the Ecuadorian Andes, at Parque Nacional Llanganates. The new species are characterized by the spiny appearance typical of several species inhabiting montane forests. Pristimantis yanezi sp. n. is most similar to Pristimantis colonensis and Pristimantis incanus but differs from both in groin coloration and by having smaller tubercles on the upper eyelids, heels, and tarsus. Pristimantis llanganati sp. n. is most similar to Pristimantis eriphus and Pristimantis chloronotus. It can be distinguished from Pristimantis eriphus by the color pattern on the scapular region and by having smaller conical tubercles on the dorsum. Pristimantis chloronotus differs from Pristimantis llanganati sp. n. in having a pair of sinuous paravertebral folds. Both new species occur in a region with few amphibian collections and nothing is known about their abundance and ecology. Therefore, it is recommended to assign them to the Data Deficient Red List category. Updated figures of species richness of Pristimantis among biogeographic regions in Ecuador are also presented. Pristimantis reach their highest diversity in Montane Forests of the eastern versant of the Andes. Its species richness across regions cannot be explained by regional area, elevation, temperature, or precipitation. Political endemism in Pristimantis is higher than that of other terrestrial vertebrates.