We provide qualitative and quantitative data on the natural history and foraging behavior of the ground-dwelling ant Odontomachus chelifer (Latreille) (Formicidae: Ponerinae) in a forest reserve in southeastern Brazil, with emphasis on colony activity rhythms and diet preferences in relation to seasonal availability of potential food items in the leaf litter. Ant colonies exhibited nocturnal activity throughout the year, and they foraged significantly more intensively in the wet/ warm (November–March) than in the cold/dry season (April–October). As the night begins, small groups of workers disperse and hunt individually on a wide diversity of litter arthropods of variable sizes. At dusk, encounters with foragers of the diurnal ponerine Pachycondyla striata Fr. Smith were conspicuously avoided by O. chelifer, which occasionally had their prey robbed by the former or were even taken as prey themselves. Termites were the preferred prey of O. chelifer, making up 40% of the food items captured in each season. Seasonal comparisons of prey organisms captured by O. chelifer, and of litter-dwelling arthropods sampled in pitfall traps, revealed that the frequency distribution of retrieved prey in each taxonomic group did not differ seasonally, despite the 2.7-fold increase in the overall availability of litter arthropods in the warm/wet period. This result supports foraging theory by showing that preference for certain animal prey types (i.e., taxonomic groups) persists through time despite seasonal fluctuations in the overall availability of potential prey on the forest floor. This study points out to the importance of studying ant foraging ecology and diet preferences in a natural context.