Diet of co-occurring Barn Owl and Spotted Eagle Owl has been studied by means of pellet contents analysis in urban and rural environments in the Highveld of South Africa. In urban environment, diet of both owl species was dominated by murid rodents (mainly Otomys, Mastomys and Rhabdomys). In rural environment, Barn Owl diet was also dominated by murid rodents, but in the diet of the Spotted Eagle Owl higher proportion of birds and non-murid rodents was recorded. Although in the rural environment the breadth of diet niche was wider in Spotted Eagle Owl (DB = 35.41) than in Barn Owl (DB = 12.67), there was almost total dietary overlap (DO = 0.98) between these two co-occurring owl species. For contrast, there was only slight food niche overlap (DO = 0.12) between these owl species co-occurring in the urban environment, but the diet breadth here was also wider in Spotted Eagle Owl (DB = 29.02) than in Barn Owl (DB = 17.90). In the urban environment diet breadth of the Spotted Eagle Owl is, therefore, slightly wider than in rural environment, while in the case of the Barn Owl the reverse is true. Probably there is lower abundance of available prey in urban and rural areas in the Highveld, in comparison with more natural habitats. This may force both species to resort to a more diverse diet to meet their energy requirements. Both species show, therefore, high plasticity of foraging.