As predators of a wide variety of livestock and wildlife species, wolves (Canis lupus) could serve as definitive hosts for species of Sarcocystis parasites infecting many intermediate hosts. Sarcocystis sporocysts have been observed in the feces of wolves fed infected beef before, but genetic tools have not yet definitively identified which species of parasites they harbor, impairing understanding of their contribution to infections in wild and domesticated intermediate hosts. Therefore, we genetically characterized sporocysts derived from a small sample of naturally infected wolves in Minnesota. Doing so established evidence that wolves may excrete sporocysts and therefore transmit Sarcocystis cruzi to cattle (Bos taurus), Sarcocystis mehlhorni to black-tailed deer (Odocoileus hemionus columbianus), and Sarcocystis wenzeli to chickens (Gallus domesticus). Given their wide host range and appetite, wolves may serve as a source of infection for cattle, deer, and chickens.