Abstract Trypanosoma comprises flagellates able to infect several mammalian species and is transmitted by several groups of invertebrates. The order Chiroptera can be infected by the subgenera Herpetosoma, Schizotrypanum, Megatrypanum and Trypanozoon. In this study, we described the diversity of bat trypanosomes and inferred phylogenetic relationships among the trypanosomes from bats caught in Tapajós-Arapiuns Extractive Reserve (Resex) in Pará state. Trypanosomes from bats were isolated by means of hemoculture, and the molecular phylogeny was based on the trypanosome barcode (SSUrDNA V7V8 variable region). A total of 111 bats were caught in the area, belonging to three families (Emballonuridae, Molossidae and Phyllostomidae) and 12 species. The bat trypanosome prevalence, as evaluated through hemoculture, was 9% all positive cultures were cryopreserve (100% of isolation success). Phylogenetic trees grouped nine isolates in T. cruzi marinkellei branch and only one in T. dionisii branch. Studies on bat trypanosome diversity are important for identifying pathogenic species and for generating support for control measures, especially in such areas where humans inhabit the forest with close contact with bat species. In addition, this is the first study in Resex Tapajós-Arapiuns extractive reserve and further studies should be conducted to elucidate the role of these parasites as environmental degradation biomarkers.