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The genus Hylodes Fitzinger is composed of diurnal frogs that live associated to lotic streams in forests (Lingnau et al. 2008; Silva & Benmaman 2008). In Brazil, this genus occurs from the states of Espírito Santo to Rio Grande do Sul (Lingnau et al. 2008; Frost 2011). Hylodes currently comprises 24 species (Frost 2011) distributed in four species groups (Heyer 1982): glaber; lateristrigatus; mertensi; and nasus. Hylodes ornatus (Bokermann) are included in “Hylodes lateristrigatus” species group with other 17 species (Silva & Benmaman 2008; Canedo & Pombal 2007). Fourteen out of 24 species of Hylodes have their larvae described (Pirani et al. 2011).

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The microhylid frogs belonging to the genus Arcovomer have been reported from lowland Atlantic Rainforest in the Brazilian states of Espírito Santo, Rio de Janeiro, and São Paulo. Here, we use DNA barcoding to assess levels of genetic divergence between apparently isolated populations in Espírito Santo and Rio de Janeiro. Our mtDNA data consisting of cytochrome oxidase subunit I (COI) nucleotide sequences reveals 13.2% uncorrected and 30.4% TIM2 + I + Γ corrected genetic divergences between these two populations. This level of divergence exceeds the suggested 10% uncorrected divergence threshold for elevating amphibian populations to candidate species using this marker, which implies that the Espírito Santo population is a species distinct from Arcovomer passarellii. Calibration of our model-corrected sequence divergence estimates suggests that the time of population divergence falls between 12 and 29 million years ago.