American mink (Neovison vison) is a widely distributed invasive species in southern Chile. Thirty four feral minks were trapped at two distinct sites (rural and peri-urban), diet analyzed and Toxoplasma gondii exposure compared using PCR and specific antibodies. Serum samples were evaluated using a commercial latex agglutination test where a titer ≥ 1:32 was considered positive. Of 30 mink analyzed, 21 (70%) were positive to T. gondii antibodies, with titers ranging from 1:32 to 1:2048. As expected, adult mink showed higher seroprevalence of exposure to T. gondii (18/21) than young mink (3/9) (P=0.008). There was not statistically significant difference between sex groups (P=0.687). Differences in seroprevalence were observed between the two sample sites with a higher proportion of positive individuals in the peri-urban area, and therefore, closer to human settlements (35.7% vs. 100%, P=0.0001). Individuals positive to T. gondii using PCR and/or serology showed similar differences by site with higher infected individuals in peri-urban areas (58.8% vs. 100%, P=0.007). Diet of American mink based in fecal composition analyses was mainly based on crustaceans (frequency of occurrence: crustaceans=100%, birds and rodents<7%), suggesting that the high observed prevalence of T. gondii infection might be more associated with its aquatic behavior (e.g. ingestion of oocysts in contaminated fresh water) than with their trophic behavior (e.g. preying over species that can have T. gondii cysts in their tissues). As an invasive species potentially subject to routine culling to maintain population sizes under control, minks could be used as a sentinel species to monitor pathogens of public and wildlife health importance, such as T. gondii, in aquatic environments.