Virulence is of central importance in host-parasite interactions, yet little is known about how it changes over extended evolutionary periods. In this study, all four species in the testacea species group of Drosophila were experimentally infected with sympatric and allopatric nematodes in the Howardula aoronymphium species complex, and the effect of parasite infection on three components of host fitness was determined. The Drosophila species show striking differences in their responses to infection, with reductions reaching 80% in adult lifespan, 100% in female fertility, and 90% in male fertility. Female sterility appears to be determined by the host; species that are sterilized by their local nematodes are also sterilized by the other allopatric nematodes in the H. aoronymphium complex. Host species that are not sterilized by their local parasite are not sterilized by other nematodes in the complex. In contrast, reductions in host adult lifespan and male fertility depend on both the host and the parasite. Whereas all nematodes reduced the survival of their local host species equally (about 40–45%), survival of two host species was drastically reduced (about 80%) when infected with an allopatric parasite. Thus, virulence is evolutionarily labile in associations between Drosophila testacea group species and their Howardula parasites. The data suggest that changes in the sterility component of virulence are due primarily to host evolution, whereas changes in the host mortality component are due in large part to parasite evolution.