Vampire bats in Belize harbor multiple Trypanosoma cruzi genotypes: implications for parasite transmission at the wildlife–domestic–human interface

Abstract Background Chagas disease, caused by Trypanosoma cruzi, is a neglected tropical disease with complex sylvatic and domestic transmission cycles involving vectors, mammalian hosts, and humans. The common vampire bat (Desmodus rotundus) is an obligate blood-feeding species that frequently interacts with livestock and humans, yet their role in parasite maintenance in Central America remains poorly characterized. Methodology/Principal Findings We analyzed 205 blood samples from vampire bats collected at two sites in northern Belize over three years (2019, 2021, 2022). PCR screening revealed an overall T. cruzi prevalence of 41.5% and increasing infection risks over time. The amplicon-based next-generation sequencing of the parasite mini-exon locus identified 36 unique haplotypes belonging to five DTUs: TcI, TcIV, TcV, TcVI, and TcBat. TcBat was present in half of the samples sequenced. TcBat and TcVI, were detected in Belize for the first time. Belizean TcBat haplotypes clustered closely with Colombian reference sequences, and a subset formed a Belize-specific clade, indicating previously unrecognized TcBat diversity in the region; in contrast, Brazilian TcBat sequences were more distantly related. Conclusions/Significance Vampire bats in Belize harbor diverse T. cruzi genotypes, including DTUs with established roles in human disease. Given their obligate blood diet, frequent feeding on livestock, and occasional biting of humans, vampire bats may serve as bridge hosts linking sylvatic, domestic, and human transmission cycles. Together with a recent report of an acute human Chagas disease case in northern Belize, these results underscore the need for integrated One Health surveillance of bats, vectors, livestock, and humans to better evaluate and mitigate the risk of Chagas disease in Central America. Author Summary 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 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, 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. Competing Interest Statement The authors have declared no competing interest.