Molecular genetics and symbiont diagnostics have revolutionized our understanding of insect species diversity, and the transformative effects of bacterial symbionts on host life history. Encarsia inaron is a parasitoid wasp that has been shown to harbor two bacterial endosymbionts, Wolbachia and Cardinium. Known then as E. partenopea, it was introduced to the USA in the late 1980s from populations collected in Italy and Israel for the biological control of an ornamental tree pest, the ash whitefly, Siphoninus phillyreae. We studied natural populations from sites in the United States, the Mediterranean and the Middle East as well as from a Cardinium-infected laboratory culture established from Italy, with the aims of characterizing these populations genetically, testing reproductive isolation, and determining symbiont infection status in their native and introduced range, and symbiont role. The results showed that the two Encarsia populations introduced to the USA are genetically distinct, reproductively isolated, have different symbionts and different host31 symbiont interactions, and can be considered different biological species. One (“E. inaron”) is doubly infected by Wolbachia and Cardinium, while only Cardinium is present in the other (“E. partenopea”). The Cardinium strains in the two species are distinct, although closely related, and crossing tests indicate that the Cardinium infecting “E. partenopea” induces cytoplasmic incompatibility. The frequency of symbiont infection found in the native and introduced range of these wasps was similar, unlike the pattern seen in some other systems. These results also lead to a retelling of a successful biological control story, with several more characters than had been initially described.

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