Abstract. AIM: This study aimed to evaluate influence of hydrological variation, ontogeny and interspecific variation in the feeding activity and diet composition for P. nattereri and S. marginatus in floodplain of Negro River, South Pantanal. METHODS: The samples were taken with the use of gillnets and cast nets of different sizes, from October/2005 to August/2008. RESULTS: We sampled 748 specimens, 442 of P. nattereri and 306 of S. marginatus. We identified 31 items in the dry and 14 in the flood season for P. nattereri, and 29 items in the dry and eight in the flood season for S. marginatus. For both species, fish was the predominant food item in both seasons. The PERMANOVA results showed that the diet varied significantly between the two species (p<0.001), during ontogenetic development (p<0.001) and seasonally (p=0.024). The ancova results suggests that the intensity of food intake was higher in the dry season (p<0.001) for both species. The niche breadth varied only between studied species with S. marginatus presenting higher niche breadth than P. nattereri. CONCLUSION: Piranhas feed more during dry season, probably because in this period food is more varied and abundant, due to the concentration of fish in the main river channel. There were ontogenetic changes in the diet, with no feeding overlap between the two species, probably because of differences in preference for some items and differences in feeding behavior displayed during hunting attacks.