Phenotypic and life-history diversification in Amazonian frogs despite past introgressions.

The advent of genomics in phylogenetics and population genetics strengthened the perception that conflicts among gene trees are frequent and often due to introgression. However, hybridization occurs mostly among species that exhibit little phenotypic differentiation. A recent study delineating species in Anomaloglossus, a frog genus endemic to the Guiana Shield, identified an intriguing pattern in the A. baeobatrachus species complex. This complex occurs in French Guiana and Amapá (Brazil) and comprises two sympatric phenotypes contrasting not only in body size, habitat, and advertisement call, but also in larval development mode (endotrophic vs exotrophic tadpoles). However, molecular and phenotypic divergences are, in some cases, incongruent, i.e specimens sharing mtDNA haplotypes are phenotypically distinct, suggesting a complex evolutionary history. Therefore, we genotyped 106 Anomaloglossus individuals using ddRADseq to test whether this phenotype/genotype incongruence was a product of phenotypic plasticity, incomplete lineage sorting, multiple speciation events, or admixture. Based on more than 16,000 SNPs, phylogenetic and population genetic approaches demonstrated that exotrophic populations are paraphyletic. Species tree and admixture analyses revealed a strikingly reticulate pattern, suggesting multiple historical introgression events. The evolutionary history of one exotrophic population in northern French Guiana is particularly compelling given that it received genetic material from exotrophic ancestors but shows very strong genetic affinity with the nearby endotrophic populations. This suggests strong selection on larval development and mating call after secondary contact and hybridization. The case of A. baeobatrachus represents a striking example of introgression among lineages that are phenotypically distinct, even in their larval development mode, and highlights how high-resolution genomic data can unravel unexpectedly complex evolutionary scenarios.

Cryptic diversity in Amazonian frogs: Integrative taxonomy of the genus Anomaloglossus (Amphibia: Anura: Aromobatidae) reveals a unique case of diversification within the Guiana Shield.

Lack of resolution on species boundaries and distribution can hamper inferences in many fields of biology, notably biogeography and conservation biology. This is particularly true in megadiverse and under-surveyed regions such as Amazonia, where species richness remains vastly underestimated. Integrative approaches using a combination of phenotypic and molecular evidence have proved extremely successful in reducing knowledge gaps in species boundaries, especially in animal groups displaying high levels of cryptic diversity like amphibians. Here we combine molecular data (mitochondrial 16S rRNA and nuclear TYR, POMC, and RAG1) from 522 specimens of Anomaloglossus, a frog genus endemic to the Guiana Shield, including 16 of the 26 nominal species, with morphometrics, bioacoustics, tadpole development mode, and habitat use to evaluate species delineation in two lowlands species groups. Molecular data reveal the existence of 18 major mtDNA lineages among which only six correspond to described species. Combined with other lines of evidence, we confirm the existence of at least 12 Anomaloglossus species in the Guiana Shield lowlands. Anomaloglossus appears to be the only amphibian genus to have largely diversified within the eastern part of the Guiana Shield. Our results also reveal strikingly different phenotypic evolution among lineages. Within the A. degranvillei group, one subclade displays acoustic and morphological conservatism, while the second subclade displays less molecular divergence but clear phenotypic divergence. In the A. stepheni species group, a complex evolutionary diversification in tadpole development is observed, notably with two closely related lineages each displaying exotrophic and endotrophic tadpoles.

Amphibians found in the Amazonian Savanna of the Rio Curiaú Environmental Protection Area in Amapá, Brazil / Anfíbios em Savana Amazônica na Área de Proteção Ambiental do Rio Curiaú, Amapá, Brasil

Abstract Amphibian research has grown steadily in recent years in the Amazon region, especially in the Brazilian states of Amazonas, Pará, Rondônia, and Amapá, and neighboring areas of the Guiana Shield. Even so, few data are available for the Amazonian savannas of Brazil. To contribute to the understanding of the diversity of the amphibians of these savannas, we surveyed 15 temporary ponds, six located in open areas, seven on the edge of savanna forest, and two within the forest, in the savanna of the Rio Curiaú Environmental Protection Area (EPA) in Macapá, in the state of Amapá, northern Brazil. Sampling occurred from May 2013 to August 2014 during periods when the ponds contained water. Amphibians were sampled through visual and auditory surveys conducted during both the day- and the nighttime periods on seven days each month over a total of 84 days. A total of 1574 individuals belonging to four families, 12 genera, and 28 species were recorded during the surveys. The cumulative species curve reached the asymptote, indicating that sampling effort was adequate. The number of species was 25% higher than that reported in other studies in Amazonian savannas. Twelve species were recorded for the first time in the savanna of Amapá. Lysapsus boliviana was the most common species (n = 332 of the individuals recorded). The greatest amphibian species richness found in the Rio Curiaú EPA was associated with the savanna mosaic, forest patches, lowland swamp, and temporary ponds. The results of this study contribute to the understanding of the diversity of amphibian species in the Guianan area of endemism in northern Brazil, and also the Amazonian savannas.

Resumo Existe um crescimento constante em estudos sobre os anfíbios Amazônicos especialmente nas regiões do Amazonas, Pará, Rondônia, Amapá e outras áreas no Escudo das Guianas. Porém, poucos estudos são realizados em Savana Amazônica no Brasil. Para diminuir esta lacuna, amostramos anfíbios em 15 lagoas temporárias, sendo seis em área aberta, sete com a borda próxima a mata de savana e 2 com borda interna em contato com a mata de savana na Área de Proteção Ambiental (APA) do Rio Curiaú em Macapá no estado do Amapá, norte do Brasil. As amostragens ocorreram de maio de 2013 a agosto de 2014, nos períodos em que as lagoas continham água, sete dias por mês, totalizando 84 dias de amostragem, através de procura visual e auditiva no período diurno e noturno. Os 1574 indivíduos registrados pertenciam a quatro famílias, 12 gêneros e 28 espécies. A curva de acumulação de espécies atingiu a assíntota, indicando que o esforço amostral foi suficiente. O número de espécies foi 25% maior que o registrado nos outros estudos em Savanas Amazônicas. Doze espécies foram registradas pela primeira vez em Savana Amapaense. Lysapsus boliviana contribuiu com o maior número de indivíduos (n=332). A maior riqueza de anfíbios nesta savana foi associada ao mosaico de savana, ilhas de matas de savana, florestas de várzea e lagoas temporárias existentes na APA do Curiaú. Os dados apresentados contribuem para preencher a lacuna de conhecimento para o Centro de Endemismo Guiana no norte do Brasil e em Savana Amazônica.

Cryptic diversity in the Hypsiboas semilineatus species group (Amphibia, Anura) with the description of a new species from the eastern Guiana Shield.

We used molecular and morphological data to investigate the hidden diversity within the Hypsiboas semilineatus species group, and more specifically within H. geographicus, an allegedly widespread species in northern South America. As a result, the identity of H. geographicus was clarified, several candidate species were detected and one of them, from the eastern Guiana Shield, is described herein as a preliminary step to resolve the taxonomy of the group. Hypsiboas diabolicus sp. nov. is mainly distinguished from closely-related species by an acuminate snout in lateral view, well-developed webbing between fingers and toes, and unspotted carmine/crimson colouration on the concealed surfaces of legs, feet and hands in life. The tadpole of the new species is described and is characterized by a large A-2 gap, a mostly single row of large marginal papillae, and a dark brown to black colouration. We also describe the advertisement call of the new species, which is defined as a soft call consisting of short clusters of 2-3 chuckles with a dominant frequency ranging between 1.11-1.19 kHz. Hypsiboas diabolicus sp. nov. is currently known only from the eastern Guiana Shield, and is probably endemic to that region. The new species' range overlaps broadly with another candidate species referred to as H. aff. semilineatus 1. Our preliminary results stress out a high cryptic diversity in that species group and the need for a formal redescription of Hypsiboas geographicus based on more topotypic material than what is currently available  to properly sort out the taxonomic status of several lineages in that clade.

Predation of a squirrel monkey (Saimiri sciureus) by an Amazon tree boa (Corallus hortulanus): even small boids may be a potential threat to small-bodied platyrrhines.

Predation has been suggested to play a major role in the evolution of primate ecology, although reports of predation events are very rare. Mammalian carnivores, raptors, and snakes are known predators of Neotropical primates, and most reported attacks by snakes are attributed to Boa constrictor (terrestrial boas). Here, we document the predation of a squirrel monkey (Saimiri sciureus) by an Amazon tree boa (Corallus hortulanus), the first record of the predation of a platyrrhine primate by this boid. The event was recorded during a nocturnal herpetological survey in the Piratuba Lake Biological Reserve, in the north-eastern Brazilian Amazon. The snake was encountered at 20:00 hours on the ground next to a stream, at the final stage of ingesting the monkey. The C. hortulanus specimen was 1620 mm in length (SVL) and weighed 650 g, while the S. sciureus was a young adult female weighing 600 g, 92 % of the body mass of the snake and the largest prey item known to have been ingested by a C. hortulanus. The evidence indicates that the predation event occurred at the end of the afternoon or early evening, and that, while capable of capturing an agile monkey like Saimiri, C. hortulanus may be limited to capturing small platyrrhines such as callitrichines.

Comparative cytogenetics of Physalaemus albifrons and Physalaemus cuvieri species groups (Anura, Leptodactylidae).

Recently, Physalaemus albifrons (Spix, 1824) was relocated from the Physalaemus cuvieri group to the same group as Physalaemus biligonigerus (Cope, 1861), Physalaemus marmoratus (Reinhardt & Lütken, 1862) and Physalaemus santafecinus Barrio, 1965. To contribute to the analysis of this proposition, we studied the karyotypes of Physalaemus albifrons, Physalaemus santafecinus and three species of the Physalaemus cuvieri group. The karyotype of Physalaemus santafecinus was found to be very similar to those of Physalaemus biligonigerus and Physalaemus marmoratus, which were previously described. A remarkable characteristic that these three species share is a conspicuous C-band that extends from the pericentromeric region almost to the telomere in the short arm of chromosome 3. This characteristic is not present in the Physalaemus albifrons karyotype and could be a synapomorphy of Physalaemus biligonigerus, Physalaemus marmoratus and Physalaemus santafecinus. The karyotype of Physalaemus santafecinus is also similar to those of Physalaemus marmoratus and Physalaemus biligonigerus owing to the presence of several terminal C-bands and the distal localization of the NOR in a small metacentric chromosome. In contrast, the Physalaemus albifrons karyotype has no terminal C-bands and its NOR is located interstitially in the long arm of submetacentric chromosome 8. The NOR-bearing chromosome of Physalaemus albifrons very closely resembles those found in Physalaemus albonotatus (Steindachner, 1864), Physalaemus cuqui Lobo, 1993 and some populations of Physalaemus cuvieri Fitzinger, 1826. Additionally, the Physalaemus albifrons karyotype has an interstitial C-band in chromosome 5 that has been exclusively observed in species of the Physalaemus cuvieri group. Therefore, we were not able to identify any chromosomal feature that supports the reallocation of Physalaemus albifrons.

Evaluation of the taxonomic status of populations assigned to Phyllomedusa hypochondrialis (Anura, Hylidae, Phyllomedusinae) based on molecular, chromosomal, and morphological approach.

BACKGROUND: The taxonomic and phylogenetic relationships of the genus Phyllomedusa have been amply discussed. The marked morphological similarities among some species hamper the reliable identification of specimens and may often lead to their incorrect taxonomic classification on the sole basis of morphological traits. Phenotypic variation was observed among populations assigned to either P. azurea or P. hypochondrialis. In order to evaluate whether the variation observed in populations assigned to P. hypochondrialis is related to that in genotypes, a cytogenetic analysis was combined with phylogenetic inferences based on mitochondrial and nuclear sequences. RESULTS: The inter- and intra-population variation in the external morphology observed among the specimens analyzed in the present study do not reflect the phylogenetic relationships among populations. A monophyletic clade was recovered, grouping all the specimens identified as P. hypochondrialis and specimens assigned P. azurea from Minas Gerais state. This clade is characterized by conserved chromosomal morphology and a common C-banding pattern. Extensive variation in the nucleolar organizing region (NOR) was observed among populations, with four distinct NOR positions being recognized in the karyotypes. Intra-population polymorphism of the additional rDNA clusters observed in specimens from Barreiras, Bahia state, also highlights the marked genomic instability of the rDNA in the genome of this group. Based on the topology obtained in the phylogenetic analyses, the re-evaluation of the taxonomic status of the specimens from the southernmost population known in Brazil is recommended. CONCLUSIONS: The results of this study support the need for a thorough revision of the phenotypic features used to discriminate P. azurea and P. hypochondrialis. The phylogenetic data presented here also contribute to an extension of the geographic range of P. hypochondrialis, which is known to occur in the Amazon basin and neighboring areas of the Cerrado savanna, where it may be sympatric with P. azurea, within contact zones. The misidentification of specimens may have led to inconsistencies in the original definition of the geographic range of P. azurea. The variability observed in the NOR of P. hypochondrialis reinforces the conclusion that these sites represent hotspots of rearrangement. Intraspecific variation in the location of these sites is the result of constant rearrangements that are not detected by classical cytogenetic methods or are traits of an ancestral, polymorphic karyotype, which would not be phylogenetically informative for this group.