The aim of this study was to inventory the species of small mammals in Uberlândia, Minas Gerais, Brazil, based on regurgitated pellets of the barn owl and to compare the frequency of rodent species in the diet and in the environment. Since in the region there is a high incidence of hantavirus infection, we also evaluate the importance of the barn owl in the control of rodents that transmit the hantavirus. Data on richness and relative abundance of rodents in the municipality were provided by the Centro de Controle de Zoonoses, from three half-yearly samplings with live traps. In total, 736 food items were found from the analysis of 214 pellets and fragments. Mammals corresponded to 86.0% of food items and were represented by one species of marsupial (Gracilinanus agilis) and seven species of rodents, with Calomys tener (70.9%) and Necromys lasiurus (6.7%) being the most frequent. The proportion of rodent species in barn owl pellets differed from that observed in trap samplings, with Calomys expulsus, C. tener and Oligoryzomys nigripes being consumed more frequently than expected. Although restricted to a single place and based on few individuals, the present study allowed the inventory of eight species of small mammals in Uberlândia. The comparison of the relative frequencies of rodent species in the diet and in the environment indicated selectivity. The second most preyed upon species was N. lasiurus, the main hantavirus reservoir in the Cerrado biome. In this way, the barn owl might play an important role in the control of this rodent in the region, contributing to the avoidance of a higher number of cases of hantavirus infection.