Didelphis marsupialis is a marsupial species that effectively adapts to synanthropic processes developing in cities. This marsupial lives closely with domestic animals and humans, which has favored the active exchange of parasites, thus increasing polyparasitism. Hence, this study aimed to determine the prevalence of helminths and protozoans infecting D. marsupialis in the Corregimiento of Santa Elena, Medellín. Twenty-three individuals were captured and classified as male, female, adult, or juvenile. The fecal samples were analyzed using various coprodiagnostic techniques. The eggs and oocysts were identified by microscopic evaluation of their morphology and morphometry. Twelve parasite species were identified: nine nematodes, one acanthocephalan, and two protozoans. In addition, an Adeleid coccidia considered pseudoparasite was found. The most prevalent parasite species (>
50%) were Eimeria sp, Cruzia sp., Aspidodera sp., and Gnathostoma turgidum, and nematode larvae. No significant differences were observed between parasite prevalence and host sex or age. Parasites of public health interest, such as Trichuris spp., Capillariidae nematodes, Strongyloides spp., and Giardia spp., were also identified. This study confirmed that the urban habitat of the opossum has a high frequency and diversity of endoparasites, some of which have been reported for the first time in Colombia.