Abstract The Pampa biogeographic province covers a mere 2% of the Brazilian territory (176,496 km²). However, it stands out as a complex and diverse ecosystem, although its mammal communities are still scarcely understood. Human activities are transforming the territory into a mosaic of agroecosystems, native and exotic forest fragments, and grasslands. Here we conducted the first investigation to determine the richness of small mammal assemblages in the region based on extensive analyses of owl pellets (Tyto furcata). Craniodental remains were studied from samples collected from 12 Semi-deciduous Seasonal Forest sites in the municipality of São Lourenço do Sul, State of Rio Grande do Sul, Southern Brazil. A total of 2,617 individuals belonging to 18 taxa were recorded, including 2 marsupials (Didelphidae; 0.42%), 2 chiropterans (Molossidae, Phyllostomidae; 0.12%), and 14 rodents (Cricetidae, Muridae, Caviidae; 99.46%). The rodent genera Oligoryzomys, Mus, Calomys, and Akodon were the most common taxa. Large samples also included poorly known taxa, such as the cricetids Bibimys, Juliomys (recording here its southernmost occurrence), Lundomys, and Wilfredomys. From a biogeographical point of view, the recorded assemblage embraces a mixture of Platan, Pampean, and Atlantic Forest elements, highlighting the role of the southernmost Brazilian hills as a wedge favoring the penetration of forest micromammals to higher latitudes. Our findings testify to the great diversity of the Pampa, but also point to a growing homogeneity and dominance of rodent species that are widespread in agroecosystems. Rapid inventories based on owl pellets emerge as a suitable, economic, non-invasive tool to document these community changes.