We studied the diet of 20 Barn Owl (Tyto alba) pairs breeding in three habitats (alfalfa fields, date plantations, and villages) in the same agricultural region in the Jordan Valley, Israel. Small mammals, particularly three rodents (Levant voles [Microtus socialis guentheri], house mouse [Mus sp.], and Tristram's jird [Meriones tristrami tristrami]), comprised 73 to 88% of the 3,544 prey items taken by Barn Owls in the three habitats. Frequencies in number and biomass of the rodent species differed among habitats. The number of bird species, their frequencies, and biomass in the diet were higher in villages than in the other two habitats, and were related to the higher diversity of birds breeding in villages. The frequency of birds in the diet was negatively correlated with distance from the village to open fields. Differences in the diet of Barn Owls among the three habitats most likely reflected differences in the distribution and abundance of the prey items in each habitat.