The diet composition and dietary patterns of the Long-eared Owl Asiootus were studied in Mediterranean agrosystems in central Crete (Greece) over the winters of 2009–2015. Overall, 2,819 prey items were recovered from 1,207 pellets, belonging to six taxa of mammals, 22 taxa of birds and four taxa of insects. Small mammals were the most common prey species, accounting for 75.8% by frequency and 79.7% by biomass, followed by birds (23.2% and 20.1%); the latter being rather an island component compared to continental regions. The House Mouse Mus musculus was the most important prey species in the owl's diet (56.3%) ahead of the Wood Mouse apodemus sylvaticus (9.51%) and the Black Rat Rattus rattus (7.9%). The species proved to be a significant rodent predator in olive groves and vineyards during winter months, selecting Wood Mouse and young Black Rat more than expected. Long-eared Owl feeding ecology studies could lead to improved land management and agricultural practices in the rural landscapes of the Mediterranean.