Starch gel protein electrophoresis of mass homogenates of Hymenolepis microstoma, H. diminuta, H. citelli, and H. nana revealed a large degree of isoenzymic variation in the 11 enzyme systems tested in these species. Whereas the majority of the 68 bands found in total were species specific, 23 were shared between at least two species, and 1 band was common to all four species. Cluster analysis suggested that H. microstoma is the most dissimilar of the four species and may have diverged the earliest from the common ancestral line, whereas H. citelli and H. nana were the two most similar species and may have diverged more recently. Hymenolepis diminuta was intermediate in position, but demonstrated closer affinities with H. citelli and H. nana than with H. microstoma.