One of the greatest challenges for biodiversity conservation is the poor understanding of species diversity. Molecular methods have dramatically improved our ability to uncover cryptic species, but the magnitude of cryptic diversity remains unknown, particularly in diverse tropical regions such as the Amazon Basin. Uncovering cryptic diversity in amphibians is particularly pressing because amphibians are going extinct globally at an alarming rate. Here, we use an integrative analysis of two independent Amazonian frog clades,Engystomopstoadlets andHypsiboastreefrogs, to test whether species richness is underestimated and, if so, by how much. We sampled intensively in six countries with a focus in Ecuador (Engystomops: 252 individuals from 36 localities;Hypsiboas: 208 individuals from 65 localities) and combined mitochondrial DNA, nuclear DNA, morphological, and bioacoustic data to detect cryptic species. We found that in both clades, species richness was severely underestimated, with more undescribed species than described species. InEngystomops, the two currently recognized species are actually five to seven species (a 150–250% increase in species richness); inHypsiboas, two recognized species represent six to nine species (a 200–350% increase). Our results suggest that Amazonian frog biodiversity is much more severely underestimated than previously thought.

We review the systematics of the Hypsiboas calcaratus species complex, a group of widely distributed Amazonian hylid frogs. A comprehensive analysis of genetic,

Genetic data in studies of systematics of Amazonian amphibians frequently reveal that purportedly widespread single species in reality comprise species complexes. This means that real species richness may be significantly higher than current estimates. Here we combine genetic, morphological, and bioacoustic data to assess the phylogenetic relationships and species boundaries of two Amazonian species of the Dendropsophus leucophyllatus species group: D. leucophyllatus and D. triangulum. Our results uncovered the existence of five confirmed and four unconfirmed candidate species. Among the confirmed candidate species, three have available names: Dendropsophus leucophyllatus, Dendropsophus triangulum, and Dendropsophus reticulatus, this last being removed from the synonymy of D. triangulum. A neotype of D. leucophyllatus is designated. We describe the remaining two confirmed candidate species, one from Bolivia and another from Peru. All confirmed candidate species are morphologically distinct and have much smaller geographic ranges than those previously reported for D. leucophyllatus and D. triangulum sensu lato. Dendropsophus leucophyllatus sensu stricto occurs in the Guianan region. Dendropsophus reticulatus comb. nov. corresponds to populations in the Amazon basin of Brazil, Ecuador, and Peru previously referred to as D. triangulum. Dendropsophus triangulum sensu stricto is the most widely distributed species; it occurs in Amazonian Ecuador, Peru and Brazil, reaching the state of Pará. We provide accounts for all described species including an assessment of their conservation status.

A new phylogeny for thePristimantis lacrimosusspecies group is presented, its species content reviewed, and three new species described from the eastern slopes of the Ecuadorian Andes. Our phylogeny includes, for the first time, samples ofP. aureolineatus,P. bromeliaceus, andP. lacrimosus. The morphology of hyperdistal subarticular tubercles is also assessed among 21 species ofPristimantis. TheP. lacrimosusspecies group is composed of 36 species distributed in the Chocó, Guiana, and Amazon regions of tropical South America with a single species reaching Central America. Ancestral area reconstruction indicates that, despite its high diversity in the Amazon region, theP. lacrimosusgroup originated in the Pacific basin, Chocó region of Ecuador and Colombia.Pristimantis amaguanaesp. nov.is most closely related toP. bromeliaceus. It differs fromP. bromeliaceusby being smaller, having transversal dark bands in the hindlimbs (absent or faint inP. bromeliaceus) and the absence of discoidal fold (present inP. bromeliaceus).Pristimantis nankintssp. nov.andP. romeroaesp. nov.are part of a clade of predominantly light-green frogs that includesP. acuminatus,P. enigmaticus,P. limoncochensis, andP. omeviridis.Pristimantis nankintssp. nov.andP. romeroaesp. nov.can be distinguished from all of them by the presence of a dark dorsolateral stripe that borders a light green band on a green background. Hyperdistal tubercles are present in all examined species of theP. lacrimosusspecies group and its sister clade. Species with hyperdistal tubercles are characterized by having relatively long terminal phalanges and narrow T-shaped expansion at the end of the terminal phalange. We discuss the phylogenetic distribution of these characters and their potential diagnostic significance.

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 clade 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 clade, 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 clade 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

TheHyloscirtuslarinopygiongroup is a clade of 16 species of large hylids that inhabit cascading Andean streams. They have brown coloration that, in most species, contrasts with bright marks. Herein morphological and genetic evidence is used to describe a new species of the group from Cordillera del Cóndor, a sub-Andean mountain chain that has phytogeographic affinities with the Guianan Tepuis. The new species is characterized by dark-brown coloration with contrasting bright orange flecks and by the presence of an enlarged and curved prepollex protruding as a spine. The new species is closely related toH.tapichalacaand an undescribed species from the southern Andes of Ecuador. The genetic distance betweenH.hillisisp. n.and its closest relative,H.tapichalaca, is 2.9% (gene 16S mtDNA). Our phylogeny and a review of recently published phylogenies show that amphibians from Cordillera del Cóndor have close relationships with either Andean or Amazonian species. Amphibians do not show the Condor-Guianan Tepuis biogeographic link that has been documented in plants.