The aim of this study was to verify the activity of some digestive enzymes in four fish species with different feeding habits. Knowledge of these enzymatic activities can help us to understand the species' digestive processes. The species chosen for this study were Ctenopharyngodon idella (herbivore), Rhamdia quelen (omnivore), Leporinus obtusidens (omnivore) and Hoplias malabaricus (carnivore). The digestive tract of these species was divided into four portions to estimate enzymatic activity: stomach, anterior, mid and posterior intestine. Ctenopharyngodon idella had the highest amylase and maltase activities in all portions of the gut, followed by L. obtusidens . The highest trypsin activity was observed in all gut portions of H. malabaricus, followed by the mid intestine of L. obtusidens and the anterior intestine of C. idella . The highest chymotrypsin activity was found in all portions of C. idella followed by the anterior intestines of R. quelen, L. obtusidens and H. malabaricus . In the stomach, acid protease activity was significantly lower in R. quelen and L. obtusidens compared to H. malabaricus. Ctenopharyngodon idella showed high activity of enzymes that hydrolyze carbohydrates, represented in this study by amylase and maltase and H. malabaricus showed higher protease activity and low amylase activity.