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During natural parasitization,  Asobara japonica  wasps introduce lateral oviduct (LO) components into their  Drosophila  hosts soon after the venom injection to neutralize its strong toxicity; otherwise, the host will die. Although the orchestrated relationship between the venom and LO components necessary for successful parasitism has attracted the attention of many researchers in this field, the molecular natures of both factors remain ambiguous. We here showed that precipitation of the venom components by ultracentrifugation yielded a toxic fraction that was inactivated by ultraviolet light irradiation, boiling, and sonication, suggesting that it is a virus-like entity. Morphological observation of the precipitate after ultracentrifugation showed small spherical heterogeneous virus-like particles 20–40 nm in diameter. The venom’s detrimental effect on  D .  melanogaster  larvae was not directly neutralized by the LO components but blocked by a hemolymphal neutralizing factor activated by the LO factor. Furthermore, we found that  A .  japonica  venom and LO components acted similarly on the larvae of the common cutworm  Spodoptera litura : the venom injection caused mortality but coinjection of the LO factor protected  S .  litura  larvae from the venom’s toxicity. In contrast,  D .  ficusphila  and  D .  bipectinata , which are closely related to  D .  melanogaster  but non-habitual host species of  A .  japonica , were not negatively affected by  A .  japonica  venom due to an intrinsic neutralizing activity in their hemolymph, indicating that these species must have acquired a neutralizer of  A .  japonica  venom during evolution. These results give new insights into the characteristics of both the venom and LO components:  A .  japonica  females have utilized the virus-like toxic venom factor to exploit a wider range of host species after the evolutionary process enabled them to use the LO factor for activation of the host hemolymph neutralizer precursor, although the non-habitual host  Drosophila  species possess an active intrinsic neutralizer in their hemolymph.

Characterization of Venom and Oviduct Components of Parasitoid Wasp Asobara japonica