In this work, we report the amphibian fauna composition of a natural protected area located near Esperanza city, Province of Santa Fe, Argentina. It opened to the public in 2015, with yet no baseline inventory of amphibian species. Additionally, we evaluated the presence of the amphibian parasitic fungus Batrachochytrium dendrobatidis (Bd). We carried out eleven samplings from October 2015 to March 2016 in three environmental units: native forest, exotic forest, and floodplain grasslands. The sampling methods we used were live pit-fall traps with drift fences, auditory encounters, and visual encounters. Sixteen species belonging to 9 genera and 4 families were recorded. October was the month with the highest specific richness. Bd-specific real-time Taqman qPCR made on skin swabs made on 11 species were all negative. Even small, the amphibian species identified in the protected area represent ca. 30% of the anurans already reported for Santa Fe, and 9% of the Argentinean batracofauna.