The drugs currently available for Chagas’disease treatment are unsatisfactory due to limited efficacy and toxic side effects, making the search for more specific pharmacological agents a priority. The components of the Trypanosoma cruzi trypanothione-dependent antioxidant system have been pointed out as potential chemotherapeutic targets for the development of more specific drugs. To work properly, this system must have a current supply of NADPH, provided by glucose-6-phosphate dehydrogenase (G6PD) and 6-phosphogluconate dehydrogenase (6PGD). Here, we compare two T. cruzi strains, Tulahuen 2 and Y, regarding growth rate, cytosolic tryparedoxin peroxidase (TcCPX) concentration and pentose phosphate pathway dehydrogenases activities. Tulahuen 2 cells show higher values as compared to the Y strain when the following parameters are compared: TcCPX concentration, resistance to H 2 O 2, growth index and G6PD activity. Different patterns of G6PD and 6PGD activities were observed among strains along the growth curve and when cells were challenged with H 2 O 2 . These data reinforce the heterogeneity within T. cruzi populations and also the importance of G6PD in protecting the parasite against reactive oxygen species.