In this study, we explored the territorial displays of Little Owls in urban and rural areas. Specifically, we compared the size of territories between urban and rural landscapes, investigating the main characteristics (e.g., land use composition, altitude) that characterize territories of the species in both types of landscapes. Overall, territories were smaller in urban than in rural areas, with a relatively different land use composition. Finally, even though the rate of territorial displays was similar between urban and rural territories, individuals used various types of structures differentially. Our findings provide new insight concerning the conservation of Little Owls in anthropized areas.