The feeding habits of apex predators such as raptors have a major influence on the local ecosystem, but their predation targets have been analyzed by visual observation that requires extremely proficient skills of taxonomic classification to identify small animals. Identifying prey animals at the species level reveals predator biology as well as important information for conserving predators, but it is difficult using conventional techniques. In this study, we aimed to clarify the diet of the long-eared owl Asio otus by DNA metabarcoding of its pellets in the central region of Kanagawa Prefecture in Japan, using three types of universal primer sets. The results indicate that the long-eared owl feeds on various types of warm-blooded animals including birds, rodents and bats in winter. Our findings also demonstrate the high efficiency and reliability of DNA metabarcoding in the study of predator–prey relationship.

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