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Abstract The identification of anuran amphibians is still a challenge in megadiverse assemblages. In the Neotropics, the Atlantic Forest harbors more than 600 anuran species, and many studies in this ecoregion report anuran assemblages surpassing 30 species. Taxonomic keys facilitate the identification of biological diversity, however only a few are available for anuran assemblages in the Atlantic Forest. Herein we present an identification key for 40 anuran species distributed across 20 genera and nine families, occurring in the Environmental Protection Area of Catolé and Fernão Velho, northeastern Atlantic Forest. Thirty-five morphological characteristics were used in the key, all of which can be easily observed in living and museum specimens. This pioneer study provides the first identification key for an amphibian assemblage in the northeastern Atlantic Forest and this baseline information acts as the starting point for the development of evolutionary and ecological research in this conservation unit.