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

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.