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Chagas disease, considered a neglected disease, is a parasitic infection caused by Trypanosoma cruzi, which is endemic throughout the world. Previously, the antimicrobial effect of Mastoparan (MP) from Polybia paulista wasp venom against bacteria was described. To continue the study, we report in this short communication the antimicrobial effect of MP against Trypanosoma cruzi. MP inhibits all T. cruzi developmental forms through the inhibition of TcGAPDH suggested by the molecular docking. In conclusion, we suggest there is an antimicrobial effect also on T. cruzi.Mastoparan from Polybia paulista wasp venom (MP) possess trypanocidal effect.MP was effective over all main forms of Trypanosoma cruzi.MP causes induction of cytoplasmic ROS and mitochondrial dysfunction.MP could interact with TcGAPDH.