Chagas disease, caused by the parasite Trypanosoma cruzi, is a major public health concern in the Americas but remains neglected in many regions, including Central America. The parasite is typically transmitted by kissing bugs, but wild mammals also serve as important hosts. Vampire bats (Desmodus rotundus) are unique because they feed exclusively on blood and frequently bite livestock, and occasionally humans, creating opportunities for parasite transmission across different environments. In this study, we screened 205 vampire bats from northern Belize and found that over 40 % were infected with T. cruzi. By sequencing parasite DNA, we discovered a surprising diversity of DTUs, including TcI, TcIV, TcV, TcVI, and TcBat. The detection of TcVI is particularly important because this DTU, associated with human infections in South America, had not been previously reported in Belize. TcBat haplotypes were highly prevalent and genetically diverse. Our results show that vampire bats are important hosts of diverse T. cruzi genotypes in Belize and may act as bridge hosts between wildlife, livestock, and human transmission cycles. Enhanced One Health surveillance across vectors, bats, domestic animals, and humans will be critical for understanding and preventing Chagas disease emergence in Central America.