Novel Bat Trypanosoma Lineages, Range Expansion of Known Species, and the Habitat- Specialist T. cruzi IV: Evidence from a Conserved Tropical Forest in Southeastern Mexico

Trypanosomatids of the Schizotrypanum clade display high ecological and evolutionary diversity, yet their sylvatic cycles remain understudied in many Neotropical biomes. We conducted the first integrative molecular survey of Trypanosoma diversity in a conserved seasonal tropical forest in the Yucatán Peninsula, Mexico. Using satellite DNA (SatDNA), miniexon PCR, and 18S rRNA sequencing, we screened vectors, including insects ( Triatoma dimidiata ), rodents, and bats. Trypanosoma cruzi was detected at high prevalence in vectors (81.8%; 95% CI: 75.07–87.38%) and moderate to high levels in rodents (46.6%; 95% CI: 36.7–56.7%) and bats (46.2%; 95% CI: 38.56–53.97%), including frugivorous, insectivorous, and hematophagous species. Discrete typing unit (DTU) analysis showed T. cruzi DTU IV (TcIV) predominating in vectors and rodents, while TcI was more frequent in bats. Phylogenetic reconstruction based on 18S rRNA revealed multiple cryptic bat-associated lineages, including sequences related to T. wauwau , T. madeirae , and Neobat clades—reported here for the first time in Mexico. Our results underscore the role of protected forests as reservoirs of trypanosomatid diversity and the relevance of multi-host surveillance to better understand transmission dynamics. This study contributes novel insights into the ecology, biogeography, and evolutionary history of Trypanosoma spp. in Mesoamerica and supports efforts to strengthen early detection of potential zoonotic threats.