Pristimantis in the Eastern Brazilian Amazon: DNA barcoding reveals underestimated diversity in a megadiverse genus.

The genus Pristimantis has the highest species diversity among all terrestrial vertebrates, with most species observed in the Andean region and the Guiana Shield. Even with the recent description of a new species, only P. latro, P. dundeei and P. zimmermanae occur in the south of the Amazon River. The lack of taxonomists specialized in the field leads to the propagation of dubious terminologies (e.g. Pristimantis sp1, Pristimantis sp2, P. aff. Fenestratus and P. gr. conspicillatus) or even misidentification of species, resulting in erroneous species distributions. In this study, we applied the Automatic Barcode Gap Discovery (ABGD) algorithm for the delimitation of candidate species and values of genetic distances using the mitochondrial marker Cytochrome Oxidase Subunit I (COI), proposed in the barcode methodology, where values greater than 10% are considered as indicative of different species. We found large genetic distances between P. latro and Pristimantis sp1 Unconfirmed Candidate Species - UCS1 (21%), and between P. altamazonicus and Pristimantis sp2 UCS2 (14%). The ABGD method recognized UCS1 and UCS2 as distinct species. Pristimantis sp. UCS1 and UCS2 in the east of the Brazilian Amazon are indicated as candidate species. We suggest greater sampling of Pristimantis sp. UCS1 and UCS2, integrating morphology and bioacoustics to solve the taxonomic status in the east of the Brazilian Amazon.