Trypanosoma cruzi infections and associated pathology in urban-dwelling Virginia opossums (Didelphis virginiana)

Trypanosoma cruzi, a zoonotic protozoan parasite, infects a wide range of mammals. The southern United States has endemic sylvatic transmission cycles maintained by several species of wildlife and domestic dogs. We hypothesized that urban-dwelling opossums (Didelphis virginiana) in South Texas are infected with T. cruzi, and that tissue pathology would be associated with infection. In 2017, we collected blood, heart tissue and anal gland secretions from 100 wild opossums across three seasons that were trapped by animal control in South Texas. In addition, anal gland tissue and intercostal muscle were collected from 43 of the 100 opossums for which time allowed the extra tissue collection. All blood, tissue, and secretion samples were screened for T. cruzi DNA using qPCR with confirmation of positive status achieved through one or more additional PCR assays, including a qPCR to determine the parasite discrete typing unit (DTU). T. cruzi DNA was detected in at least one tissue of 15% of the opossums sampled: blood clot (9%), heart tissue (10%), anal gland secretions (12%), intercostal muscle (16.3%), and anal gland tissue (11.6%). Infection was detected in two or more different tissue types in nine of the opossums. The 35 tissues for which parasite DTU was determined were exclusively ‘Tcl’- a DTU previously associated with locally-acquired human disease in the United States. T. cruzi-positive opossums were nearly 14 times more likely to exhibit significant heart lesions on histopathology (lympoplasmacytic inflammation±fibrosis) when compared to negative opossums (OR = 13.56, CI = 1.23–751.28, p-value = 0.03). Three triatomines were opportunistically collected from the study site, of which two were infected (66.7%), and bloodmeal analysis revealed canine, opossum, and human bloodmeals. Given the presence of parasite in opossum blood, unique potential for shedding of parasite in anal glad secretions, and evidence of vectors feeding on opossums, it is likely that opossums serve as wild reservoirs around urban dwellings in South Texas.