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The small didelphid marsupial Cryptonanus guahybae was described as a subspecies of Gracilinanus microtarsus and was so far considered endemic to the state of Rio Grande do Sul, southern Brazil. We used morphological data to identify Cryptonanus specimens from the states of Santa Catarina and Paraná, deposited in two Brazilian mammal collections. The cytochrome b sequences of our samples formed a well-supported monophyletic group with low intraspecific genetic divergence. This clade showed high interspecific genetic divergence regarding sequences of the two other species of Cryptonanus that occur in Brazil, C. agricolai and C. chacoensis. Diagnostic morphological characters, such as reddish brown dorsal fur, mammary count (7-1-7=15), small molars, and the presence of a complete anterior cingulum on M3, confirm the identification of 20 specimens from southern Brazil as C. guahybae. Considering the morphological diagnoses and the monophyletic and divergent condition of the cytochrome b sequences, it is suitable that these represent the first molecular data for the species. Our results allowed the expansion of the known distribution of C. guahybae throughout the Atlantic Forest phytophysiognomies of the states of Santa Catarina and Paraná. [ABSTRACT FROM AUTHOR]