A total of 22 TVVs were identified in 10 T. vaginalis cultures, with multiple TVVs detected in nine cultures (Table 2). The higher prevalence of T. vaginalis cultures with either TVV1 or TVV2 than with TVV3 or TVV4 is consistent with previous reports [10, 37]. In these publications, concurrent TVV infection, with at least two or 3 TVVs, was recorded in six and three T. vaginalis sample cultures, respectively. Although we identified single actin genotypes in all T. vaginalis cultures, which is indicative for the presence of only 1 T. vaginalis strain per culture, we cannot rule out that the presence of the multiple TVVs may be the result of a mixture of T. vaginalis strains, each infected with a different TVV. Therefore, our data does not necessarily indicate concurrent infection of TVVs in a single TV strain. The lytic cycle of TVVs is yet to be described, and attempts to infect uninfected isolates have been unsuccessful . Therefore, it is plausible that the virus may solely be acquired through vertical transmission, making its presence an important genetic marker .