Voles are the most common vertebrate pests in European agriculture. Identifying their distribution and abundance patterns provides valuable information for future management. Barn Owl diet analysis is one of the optimum methods used to record small mammal distribution patterns on large spatial scales. From 2003 to 2005, a total of 10,065 Barn Owl pellets were collected and analyzed from 31 breeding sites in the largest agroecosystem in Greece, the Thessaly plains. A total of 29,061 prey items were identified, offering deep insight into small mammal distribution, specifically voles. Four discrete vole species (Harting’s vole Microtus hartingi, East European vole Microtus levis, Thomas’s pine vole Microtus thomasi, and Grey dwarf hamster Cricetulus migratorius) comprised 40.5% (11,770 vole prey items) of the total Barn Owl prey intake. The presence and abundance of the voles varied according to underlying environmental gradients, with soil texture and type playing a major role. M. levis showed no significant attachments to gradients, other than a mild increase in Mollisol soils. It was syntopic in all sites with M. hartingi, which was the dominant and most abundant small mammal species, preferring non-arable cultivated land, natural grasslands, set-aside fields, and fallow land. M. thomasi was strictly present in western Thessaly and strongly associated with a sandy-clay soil texture and Alfisol soils. C. migratorius was the least represented vole (162 items), exclusively present in eastern Thessaly and demonstrating a stronger association with cereals, Mollisol soils, and an argillaceous-clay soil texture. This is the first study in Greece at such a large spatial scale, offering insights for pest rodents’ distribution in intensive agroecosystems and their response to environmental gradients including soil parameters.