The biosystematic status of species belonging to the Rhipicephalus sanguineus group is difficult to determine by phenotypic methods and has been subject to frequent revisions and ongoing debate. Yet, the differentiation of species within this complex is of clinical importance especially in veterinary medicine due to different host specificities of the transmitted pathogens. In order to generate quantifiable and comparable data independent of environmental influences, a section of the second internal transcribed spacer (ITS2) of the rRNA gene was sequenced for 6 species belonging to the Rhipicephalus sanguineus complex: Rhipicephalus sanguineus sensu stricto, R. turanicus, R. rossicus, R. pumilio, R. pusillus, and R. camicasi. Rhipicephalus sanguineus s. str. and R. turanicus as well as R. pumilio and R. rossicus were found to have a close genetic relationship compatible with possible conspecificities. Conversely, R. pusillus was found to have the most distant relationship to the other species.