Abstract: The olive fruit fly Bactrocera oleae is one of the most serious and economically damaging insects worldwide, affecting the quality and quantity of both olive oil and table olives. Third instar larvae and pupae of several tephritid flies were reported to be susceptible to entomopathogenic nematodes (EPNs), but few studies have been carried out on the olive fruit fly. Laboratory assays were conducted to evaluate the susceptibility of B. oleae larvae and pupae to two commercial EPN species, Steinernema feltiae and Heterorhabditis bacteriophora and two indigenous Italian strains of H. bacteriophora and Steinernema carpocapsae. Moreover, an olive assay in the soil was performed to evaluate the capability of EPN strains to enter inside the olive fruits and deal with B. oleae during the pupal stage or the emergence of adults in the winter diapause. The susceptibility assays with B. oleae were performed in well plates, filled with sterile soil (n = 30 for each EPN strain and insect stage). Adult emergence and mortality were recorded daily for 15 days. Dead pupae and adults were dissected to assess nematode infection. The most noteworthy result was obtained with S. feltiae which was able to infect more than 80% of larvae and it killed the pupae inside olive fruits and the adults during their emergence with the same efficacy. Since this tephritid fly spent several months in the soil, the use of EPNs, in particular S. feltiae could be a promising method to control this pest.