The Serra Dona Francisca is in the subtropical region of the Atlantic Forest and part of its area is protected by a sustainable use conservation unit. In the present study, we performed an anuran inventory at different elevations, providing information on natural history of each species. Field work was carried out from October 2012 to January 2014. We sampled the anuran by visual and auditory surveys and used three 50-meter pitfall lines composed of four 60L buckets with drift-fences at each elevation area. We registered 63 species belonging to 13 families: Brachycephalidae (4), Bufonidae (4), Centrolenidae (1), Craugastoridae (1), Cycloramphidae (2), Hemiphractidae (2), Hylidae (30), Hylodidae (2), Leptodactylidae (10), Microhylidae (2), Odontophrynidae (3), Phyllomedusidae (1), and Ranidae (1). We observed four new records for the northeastern region of the state of Santa Catarina: Bokermannohyla luctuosa, Scinax aromothyella, Odontophrynus americanus, and Leptodactylus plaumanni. We registered a higher number of species during the hot and wet months, with subsequent drop during the cold months. We found an increase of species richness at higher elevations. Considering that our results represent 50% of anuran richness of the state of Santa Catarina, this is an important region for conservation of biodiversity of southern Atlantic Forest.