An elusive giant: A new species of Vitreorana Guayasamin et al., 2009 (Anura: Centrolenidae) from the northern Atlantic Forest with an osteological description and comments on integumentary spicules.

We describe a new species of Vitreorana from the Atlantic Forest of southern Bahia state, in north-eastern Brazil. Vitreorana assuh sp. nov. is by far the largest species of the genus (snout-to-vent length [SVL] = 30.9 mm and 34.1 mm in one male and female, respectively; maximum SVL recorded for all other species = 28.0 mm). The new species is morphologically most similar to V. franciscana, from the Cerrado biome in Brazil. Besides its large size, the new species has, among other diagnostic characters, the presence of vomerine teeth and the occurrence of enameled iridophores in the pericardium, hepatic and urinary bladder peritonea. Furthermore, we describe and compare the complete mineralized skeleton of the new species and V. uranoscopa, and describe and discuss the variation of integumentary spicules in all species of the genus and their relevance to Vitreorana systematics.

Natural history predicts patterns of thermal vulnerability in amphibians from the Atlantic Rainforest of Brazil.

In the Brazilian Atlantic Rainforest (AF), amphibians (625 species) face habitat degradation leading to stressful thermal conditions that constrain animal activity (e.g., foraging and reproduction). Data on thermal ecology for these species are still scarce. We tested the hypothesis that environmental occupation affects the thermal tolerance of amphibian species more than their phylogenetic relationships. We evaluated patterns of thermal tolerance of 47 amphibian species by assessing critical thermal maxima and warming tolerances, relating these variables with ecological covariates (e.g., adult macro- and microhabitat and site of larval development). We used mean and maximum environmental temperature, ecological covariates, and morphological measurements in the phylogenetic generalized least squares model selection to evaluate which traits better predict thermal tolerance. We did not recover phylogenetic signal under a Brownian model; our results point to a strong association between critical thermal maxima and habitat and development site. Forest species were less tolerant to warm temperatures than open area or generalist species. Species with larvae that develop in lentic environment were more tolerant than those in lotic ones. Thus, species inhabiting forest microclimates are more vulnerable to the synergistic effect of habitat loss and climate change. We use radar charts as a quick evaluation tool for thermal risk diagnoses using aspects of natural history as axes.

The phylogeny of Dendropsophini (Anura: Hylidae: Hylinae).

The relationships of the hyline tribe Dendropsophini remain poorly studied, with most published analyses dealing with few of the species groups of Dendropsophus. In order to test the monophyly of Dendropsophini, its genera, and the species groups currently recognized in Dendropsophus, we performed a total evidence phylogenetic analysis. The molecular dataset included sequences of three mitochondrial and five nuclear genes from 210 terminals, including 12 outgroup species, the two species of Xenohyla, and 93 of the 108 recognized species of Dendropsophus. The phenomic dataset includes 46 terminals, one per species (34 Dendropsophus, one Xenohyla, and 11 outgroup species). Our results corroborate the monophyly of Dendropsophini and the reciprocal monophyly of Dendropsophus and Xenohyla. Some species groups of Dendropsophus are paraphyletic (the D. microcephalus, D. minimus, and D. parviceps groups, and the D. rubicundulus clade). On the basis of our results, we recognize nine species groups; for three of them (D. leucophyllatus, D. microcephalus, and D. parviceps groups) we recognize some nominal clades to highlight specific morphology or relationships and facilitate species taxonomy. We further discuss the evolution of oviposition site selection, where our results show multiple instances of independent evolution of terrestrial egg clutches during the evolutionary history of Dendropsophus.

The phylogeny of the Casque-headed Treefrogs (Hylidae: Hylinae: Lophyohylini).

The South American and West Indian Casque-headed Treefrogs (Hylidae: Hylinae: Lophyohylini) include 85 species. These are notably diverse in morphology (e.g. disparate levels of cranial hyperossification) and life history (e.g. different reproductive modes, chemical defences), have a wide distribution, and occupy habitats from the tropical rainforests to semiarid scrubland. In this paper, we present a phylogenetic analysis of this hylid tribe based on sequence fragments of up to five mitochondrial (12S, 16S, ND1, COI, Cytb) and six nuclear genes (POMC, RAG-1, RHOD, SIAH, TNS3, TYR). We included most of its species (> 96%), in addition to a number of new species. Our results indicate: (i) the paraphyly of Trachycephalus with respect to Aparasphenodon venezolanus; (ii) the nonmonophyly of Aparasphenodon, with Argenteohyla siemersi, Corythomantis galeata and Nyctimantis rugiceps nested within it, and Ap. venezolanus nested within Trachycephalus; (iii) the polyphyly of Corythomantis; (iv) the nonmonophyly of the recognized species groups of Phyllodytes; and (v) a pervasive low support for the deep relationships among the major clades of Lophyohylini, including C. greeningi and the monotypic genera Itapotihyla and Phytotriades. To remedy the nonmonophyly of Aparasphenodon, Corythomantis, and Trachycephalus, we redefined Nyctimantis to include Aparasphenodon (with the exception of Ap. venezolanus, which we transferred to Trachycephalus), Argenteohyla, and C. galeata. Additionally, our results indicate the need for taxonomic work in the following clades: (i) Trachycephalus dibernardoi and Tr. imitatrix; (ii) Tr. atlas, Tr. mambaiensis and Tr. nigromaculatus; and (iii) Phyllodytes. On the basis of our phylogenetic results, we analyzed the evolution of skull hyperossification and reproductive biology, with emphasis on the multiple independent origins of phytotelm breeding, in the context of Anura. We also analyzed the inter-related aspects of chemical defences, venom delivery, phragmotic behaviour, co-ossification, and prevention of evaporative water loss.

A new species of Scinax Wagler (Hylidae: Scinaxini) from the tropical forests of Northeastern Brazil.

We describe a new species of the Scinax ruber clade from Northeastern Brazil that occurs in widely separated geographic areas in the Atlantic Forest of southern Bahia state and the Highland Humid Forest of Serra de Baturité, northeast Ceará state. Scinax tropicalia sp. nov. (holotype coordinates: -14.795694°, -39.172645°) is diagnosed from all 75 currently recognize species of the S. ruber clade by bioacoustical and morphological adult traits, such as duration (0.11-0.31 s) and dominant frequency (1.59-1.85 kHz) of the advertisement call, snout shape rounded, nearly rounded, or semi-circular in dorsal view and rounded to slightly protruding in profile, bilobate vocal sac, absence of pectoral glands and spicule-shaped papillary epidermal projections on nuptial pads, and color pattern on the dorsum of body and hidden surfaces of hindlimbs.

Chromosome evolution in Lophyohylini (Amphibia, Anura, Hylinae).

The hyline tribe Lophyohylini includes 87 species of treefrogs, of which cytogenetics aspects have been studied in less than 20% of them. In order to evaluate the evolution of some of its chromosome characters (NOR position, C-bands, and DAPI/CMA3 bands), we studied the karyotypes of 21 lophyohylines, 16 of them for the first time, and analyzed them in a phylogenetic context. Most species showed similar karyotypes regarding chromosome number (2n = 24) and morphology (FN = 48), excepting Phyllodytes edelmoi and Osteocephalus buckleyi with 2n = 22 (FN = 44) and 2n = 28 (FN = 50), respectively. The NOR location was variable among species and provided valuable phylogenetic information. This marker was located in pair 11 in all species of Trachycephalus, Itapotihyla langsdorffii, and Nyctimantis arapapa, representing the plesiomorphic condition of Lophyohylini. Besides, other apomorphic states were recovered for the clades comprising N. rugiceps and N. siemersi (NOR in pair 5), and Dryaderces pearsoni, Osteocephalus, and Osteopilus (NOR in pair 9). Phyllodytes presented variation for NORs position; they were in pair 2 in P. edelmoi, pair 7 in P. melanomystax, and pair 8 in P. gyrinaethes and P. praeceptor. Polymorphisms in size, number, and activity of this marker were observed for N. siemersi, Osteocephalus fuscifacies, and some species of Trachycephalus. Remarkably, in N. siemersi NORs were detected on a single chromosome in the two specimens studied by this technique, raising the question of how this complex polymorphism is maintained. Interstitial telomeric sequences were found in P. edelmoi, P. melanomystax, and Osteocephalus buckleyi, and their presence seems to be not related to the chromosome reorganization events. Finally, some species showed spontaneous rearrangements, possibly as a consequence of an uncommon phenomenon in anuran cytogenetics: the presence of fragile sites or secondary constrictions not associated with NORs. We propose that this rare feature would have played an important role in the evolution of this group of frogs. From the evidence obtained in this and previous studies, we conclude that Lophyohylini presents a complex chromosome evolution.

Larval anatomy of Dendropsophus decipiens (A. Lutz 1925) (Anura: Hylidae: Dendropsophini) with considerations to larvae of this genus.

The Dendropsophus decipiens clade comprises four species: D. berthalutzae, D. decipiens, D. haddadi, and D. oliveirai. Tadpoles of these species were described, but data on their internal morphology are lacking. We provide the first description of the buccopharyngeal anatomy, chondrocranial morphology, and cranial, hyoid and hyobranchial musculature of the tadpole of D. decipiens. Larvae of D. decipiens are characterized by the absence of lingual papillae, presence of fan-like papilla on the buccal floor, presence of a single-element suprarostral cartilage, presence of a small triangular process at the basis of the processus muscularis, m. levator mandibulae lateralis inserted on the nasal sac, and m. subarcualis rectus II-IV with a single, continuous slip. Tadpoles are likely macrophagous, although not as specialized as those of other species of the genus, suggesting some degree of diversification on the feeding habits within Dendropsophus.

On the identity of Adelophryne pachydactyla Hoogmoed, Borges, and Cascon, 1994 (Brachycephaloidea: Eleutherodactylidae).

The use of molecular data for documenting biodiversity has become more common over time as larger datasets can be generated faster. Nevertheless, studies addressing phenotypical data have not become as common. As a collateral effect, many samples used in molecular studies are assigned to a species without checking phenomic characters of the vouchers. Correct specific assignment is paramount for any biological hypothesis. A recent review of Phyzelaphryninae, while producing interesting results, has not specified how some vouchers used in molecular analyses were assigned to each species. Using new sequences from fresh material collected in Southern Bahia, we re-identify Adelophryne specimens based on molecular and morphological grounds. This new taxonomic understanding clarifies the phylogenetic position of A. pachydactyla and highlights a more parsimonious explanation for the evolution of a morphological character-the reduction of a phalange on Finger IV-within Adelophryne.

Another new species of Phyllodytes (Anura: Hylidae) from the Atlantic Forest of northeastern Brazil.

A new species of the genus Phyllodytes is described from the State of Bahia, in the Atlantic Rain Forest of Northeastern Brazil. Phyllodytes praeceptor sp. nov. can be differentiated from other species of Phyllodytes by its medium size (SVL 20.7-25.8 mm in males); odontoids moderately developed; vocal sac externally visible; eyes large and prominent; dorsum homogenously cream, except for a few scattered spots and blotches; venter areolate with two parallel, paramedial lines of larger tubercles; few tubercles in the ventral surface of thighs, the largest being the medial one; a large tubercle on the skin around the tibio-tarsal articulation; nuptial pad rounded and moderately expanded.

The tadpole of Sphaenorhynchus caramaschii, with comments on larval morphology of Sphaenorhynchus (Anura: Hylidae).

We describe the tadpole of Sphaenorhynchus caramaschii. It differs from tadpoles of other species of Sphaenorhynchus in having a short spiracle, submarginal papillae, and alternating short and large marginal papillae in the oral disc. Some larval characteristics, like morphology and position of the nostrils, length of the spiracle, and size of the marginal papillae on the oral disc are discussed for tadpoles of other species of Sphaenorhynchus.  

A new "Bat-Voiced" species of Dendropsophus Fitzinger, 1843 (Anura, Hylidae) from the Amazon Basin, Brazil.

We describe Dendropsophus ozzyi sp. nov., a new species of treefrog, tentatively included in the Dendropsophus microcephalus Group and most notably diagnosed by the presence of pointed fingers and an advertisement call with a very high dominant frequency. The new species is known from three localities in the Brazilian Amazon forest, two on western State of Pará and one (the type locality) in eastern State of Amazonas (03°56'50"S and 58°26'36"W, 45 m a.s.l.).

High levels of diversity uncovered in a widespread nominal taxon: continental phylogeography of the neotropical tree frog Dendropsophus minutus.

Species distributed across vast continental areas and across major biomes provide unique model systems for studies of biotic diversification, yet also constitute daunting financial, logistic and political challenges for data collection across such regions. The tree frog Dendropsophus minutus (Anura: Hylidae) is a nominal species, continentally distributed in South America, that may represent a complex of multiple species, each with a more limited distribution. To understand the spatial pattern of molecular diversity throughout the range of this species complex, we obtained DNA sequence data from two mitochondrial genes, cytochrome oxidase I (COI) and the 16S rhibosomal gene (16S) for 407 samples of D. minutus and closely related species distributed across eleven countries, effectively comprising the entire range of the group. We performed phylogenetic and spatially explicit phylogeographic analyses to assess the genetic structure of lineages and infer ancestral areas. We found 43 statistically supported, deep mitochondrial lineages, several of which may represent currently unrecognized distinct species. One major clade, containing 25 divergent lineages, includes samples from the type locality of D. minutus. We defined that clade as the D. minutus complex. The remaining lineages together with the D. minutus complex constitute the D. minutus species group. Historical analyses support an Amazonian origin for the D. minutus species group with a subsequent dispersal to eastern Brazil where the D. minutus complex originated. According to our dataset, a total of eight mtDNA lineages have ranges >100,000 km2. One of them occupies an area of almost one million km2 encompassing multiple biomes. Our results, at a spatial scale and resolution unprecedented for a Neotropical vertebrate, confirm that widespread amphibian species occur in lowland South America, yet at the same time a large proportion of cryptic diversity still remains to be discovered.

Description and phylogenetic relationships of a new species of treefrog of the Dendropsophus leucophyllatus group (Anura: Hylidae) from the Amazon basin of Colombia and with an exceptional color pattern.

We describe Dendropsophus manonegra sp. nov. from the upper Amazon basin in the eastern foothills of the Andes in Colombia (1 degree 47' 42.2" N, 75 degrees 38' 48.7" W; 1040 m a.s.l.). Phylogenetic analysis of DNA sequences of 2582 aligned base pairs of the 12S and 16S rRNA genes recovered the new species as a member of the D. leucophyllatus species group and sister to D. bifurcus. Morphological traits, such as the presence of pectoral glands in males and females, support this hypothesis. The new species is readily distinguished from all other members of the species group by having bluish-black coloration on fingers, toes, webbing, axillary membranes, groin and hidden surfaces of arms and legs. Some aspects on the composition and systematics of the D. leucophyllatus group are discussed.

Phenotypic variation of Leptodactylus cupreus Caramaschi, São-Pedro and Feio, 2008 (Anura, Leptodactylidae).

This study describes for the first time the female of Leptodactylus cupreus and provides new information concerning its geographical distribution, male's morphology and bioacustics. Leptodactylus cupreus, a poorly known species from the Brazilian Atlantic Forest, was originally allocated in the L. mystaceus complex of the L. fuscus species group. Based on morphological observations, we infer that L. cupreus should be in fact related to L. mystacinus, a species that, although assigned to the L. fuscus species group, is not assigned to the L. mystaceus complex. Therefore, we comment the phylogenetic relationships concerning L. cupreus, L. mystaceus and L. mystacinus.

From Amazonia to the Atlantic forest: molecular phylogeny of Phyzelaphryninae frogs reveals unexpected diversity and a striking biogeographic pattern emphasizing conservation challenges.

Documenting the Neotropical amphibian diversity has become a major challenge facing the threat of global climate change and the pace of environmental alteration. Recent molecular phylogenetic studies have revealed that the actual number of species in South American tropical forests is largely underestimated, but also that many lineages are millions of years old. The genera Phyzelaphryne (1 sp.) and Adelophryne (6 spp.), which compose the subfamily Phyzelaphryninae, include poorly documented, secretive, and minute frogs with an unusual distribution pattern that encompasses the biotic disjunction between Amazonia and the Atlantic forest. We generated >5.8 kb sequence data from six markers for all seven nominal species of the subfamily as well as for newly discovered populations in order to (1) test the monophyly of Phyzelaphryninae, Adelophryne and Phyzelaphryne, (2) estimate species diversity within the subfamily, and (3) investigate their historical biogeography and diversification. Phylogenetic reconstruction confirmed the monophyly of each group and revealed deep subdivisions within Adelophryne and Phyzelaphryne, with three major clades in Adelophryne located in northern Amazonia, northern Atlantic forest and southern Atlantic forest. Our results suggest that the actual number of species in Phyzelaphryninae is, at least, twice the currently recognized species diversity, with almost every geographically isolated population representing an anciently divergent candidate species. Such results highlight the challenges for conservation, especially in the northern Atlantic forest where it is still degraded at a fast pace. Molecular dating revealed that Phyzelaphryninae originated in Amazonia and dispersed during early Miocene to the Atlantic forest. The two Atlantic forest clades of Adelophryne started to diversify some 7 Ma minimum, while the northern Amazonian Adelophryne diversified much earlier, some 13 Ma minimum. This striking biogeographic pattern coincides with major events that have shaped the face of the South American continent, as we know it today.