In this study, we analysed a total of 691 pellets of the Long-eared Owl (Asio otus L.) collected from Edirne (Suburban), Istanbul (Suburban) and Kars (High altitude steppe) provinces in January-February 2019. Dietary contents of the Long-eared Owl were considering the different habitat (steppe and suburban) types. Small mammals constituted the majority of the diet content in all areas, but a small amount of bird remains were also found in pellets. 1474 prey items belonging to 7 different mammal taxa (Apodemus sp., Cricetulus sp., Crocidura sp., Micromys sp., Microtus sp., Mus sp., Rattus sp.) were identified. High amount of Mus sp. was found in pellets collected from Edirne (50.34%) and Istanbul (41.42%). On the other hand, Microtus sp. was the main prey species in Kars. Overall, our study supported that the Long-eared Owl acts as an opportunistic predator and change its dietary contents according to different environmental conditions. Incompatible results were obtained between the trapping study and pellet examination. The reasons could be that some mammal species can be caught by chance, trapping area cannot represent the entire hunting area and some mammal species could avoid trapping.