We estimated the genetic variability of nine fish species from the Brazilian upper Paraná River floodplain (Astyanax altiparanae, Hoplias malabaricus, Leporinus lacustris, Loricariichthys platymetopon, Parauchenipterus galeatus, Pimelodus maculatus, Rhaphiodon vulpinus, Roeboides paranensis and Serrasalmus marginatus) based on data for 36 putative allozyme loci obtained using corn starch gel electrophoresis of 13 enzymatic systems: aspartate aminotransferase (EC 2.6.1.1), acid phosphatase (EC 3.1.3.2), esterase (EC 3.1.1.1), glycerol-3-phosphate dehydrogenase (EC 1.1.1.8), glucose-6-phosphate dehydrogenase (EC 1.1.1.49), glucose-6-phosphate isomerase (EC 5.3.1.9), Iditol dehydrogenase (EC 1.1.1.14), isocitrate dehydrogenase - NADP+ (EC 1.1.1.42), L-lactate dehydrogenase (EC 1.1.1.27), malate dehydrogenase (EC 1.1.1.37), malate dehydrogenase-NADP+ (EC 1.1.1.40), phosphoglucomutase (EC 5.4.2.2) and superoxide dismutase, (EC 1.15.1.1). The mean expected heterozygosity varied from zero to 0.147. When data from the literature for 75 species of tropical fish were added to the nine species of this study, the heterozygosity values differed significantly among the groups of different reproductive strategies. The highest mean heterozygosity was for the non-migratory without parental care, followed by the long-distance migratory, and the lowest mean was for the non-migratory with parental care or internal fecundation.