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Background: The long-eared owl (Asio otus) has a Holarctic distribution, including much of North America. This nocturnal species is considered to be extremely secretive, and poorly known in the Great Plains of the United States and Canada, as well as to Mexico, where no previous studies on its diet have been conducted.

Findings: We analyzed 120 pellets collected during January 2007 in roosts in a 2-3 m height mesquite scrub within a grassland area of Reserva Ecologica El Uno, located in the Natural Protected Area Janos. We registered and identified three orders, four families, eight genera and ten species of mammals and two orders and one family of insects. Winter diet is dominated by mammals, especially rodents in both frequency and biomass. Cricetidae and Perognathus flavus were the most frequent family and species, respectively. On the other hand, when analyzing biomass, Sigmodon species were dominant, achieving almost 70% of the consumed biomass. Levin’s standardized niche breath based on frequency was calculated as 0.40, while based on biomass was 0.38. Also, two previously unrecorded rodent species were identified as long-eared owl prey.

Conclusion: Although 18 different types of items were identified, the long-eared owl tends to be selective, with a single genera (Sigmodon) comprising almost 70% of its consumed biomass during winter. Perognathus flavus was also important in frequency (21%); however, it barely constitutes 2% of the consumed biomass.