Abstract: Spiroplasmas were observed in seven species of the family Tabanidae (horse flies and deer flies). This is the fifth family of the order Diptera now known to harbor spiroplasmas. Noncultivable spiroplasmas were seen in the hemolymph of three species of the genus Tabanus, and cultivable forms were isolated from the guts of six species in three genera. Isolates from T. calens and T. sulcifrons were serologically similar and closely related to a spiroplasma in the lampyrid beetle, Ellychnia corrusca. These three isolates represent a new serogroup. Isolates from Hybomitra lasiophthalma were related to Group IV strains, while those from T. nigrovittatus and Chrysops sp. both represented new serogroups. At least some tabanids probably acquire spiroplasmas from contaminated flower surfaces. The possibility of vertebrate reservoirs for some tabanid spiroplasmas remains an open question.