Genetic variability was studied in five fish species (Anostomidae): Schizodon intermedius and S. nasutus and Leporinus friderici, L. elongatus and L. obtusidens, collected at one location on the Tibagi River (Paraná, Brazil). The protein data from seven systems coded collectively for 19 loci in the liver, muscle and heart. Nine of these loci were polymorphic. The estimated proportion of polymorphism loci (<img src="http:/img/fbpe/gmb/v22n4/si_p_.gif" align="absmiddle">) varied from 16.7% in S. intermedius to 36.9% in L. friderici; the mean heterozygosity observed (<img src="http:/img/fbpe/gmb/v22n4/si_h_.gif" alt="si_h_.gif (76 bytes)" align="absmiddle">o) was 0.027 ± 0.015 and 0.109 ± 0.042, respectively. The estimated value of the genetic identity among L. friderici and S. intermedius (0.749) and S. nasutus (0.787) suggested that these are "congeneric" species. Morphological characteristics indicate that these species belong to distinct genera, while isoenzymatic data show that they are very similar at the genetic/biochemical level.