Species within the genus Bulinus are responsible for transmission of schistosomes within the Schistosoma haematobium group. In order to provide a molecular insight into the species relationships within the genus, genetic variation between species representing the four species groups was assayed by Polymerase Chain Reaction (PCR) amplification of the ribosomal Internal Transcribed Spacer (ITS) region followed by Restriction Fragment Length Polymorphism (RFLP) analysis of this product with six restriction enzymes. This PCR-RFLP methodology detected considerable variation within the ITS region indicating that restriction profiles will be useful as genetic markers for identification purposes. The complete ITS1 spacer was sequenced for B. globosus, B. cernicus and B. truncatus. There were numerous nucleotide differences between taxa mainly insertions and deletions. Nucleotide divergence was calculated between species from the restriction profiles: the B. truncatus/tropicus complex and B. reticulatus group were most similar which were in turn more closely related to the B. africanus group than to the B. forskalii group. The nucleotide divergence between the species groups is substantial and questions the placement of these groups within the same genus.