The eastern portion of the state of Mato Grosso do Sul is considered a priority region for biodiversity conservation in Brazil. It lacks, however, not only large protected areas but also knowledge about wildlife species inhabiting these landscapes. This study was conducted on a farm composed of a mosaic of Eucalyptus plantations and remnants of Atlantic forest and Cerrado (Savannah) vegetation, located in the municipality of Três Lagoas. Over nine years of monitoring, 69 mammal species were recorded, including 23 bats, 14 nonvolant small-sized, and 32 medium or large-sized mammals. Twelve species are considered under a category of extinction threat. The results of this study contribute to the diagnostic of the local fauna and show the importance of this farm for mammal conservation in this region.