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The pupae of Bactrocera oleae (Diptera: Tephritidae) complete their development during autumn and winter in the soil, rather than in the drupe, resulting susceptible to edaphic predators. Environmentally friendly methods to control this olive pest involve the identification of its natural enemies. This study evaluated the role of Ocypus olens (Coleoptera: Staphylinidae) in the predation of B. oleae pupae, by means of molecular gut content analysis. Modified dry pitfall traps were used to collect live specimens from low-input olive orchards in Tuscany (Italy). Sampling was fine-tuned with a degree-day model estimating the presence of pest pupae in the soil. PCR analyses carried out on field-collected specimens demonstrated that O. olens is a predator of B. oleae, at least during autumn. These results are consistent with predictions of the degree-day model. Knowledge on species composition, traits and complementarity of the natural enemies of B. oleae pupae needs further investigation to advance conservation biological control strategies. [ABSTRACT FROM AUTHOR]