Toxoplasma gondii infection was diagnosed in 2 captive Patagonian maras (Dolichotis patagonum). One animal developed fatal systemic toxoplasmosis and had concurrent localized bacterial and fungal infections; its daughter remained clinically healthy. Microscopic findings included acute, coagulative necrosis, lymphohistiocytic inflammatory infiltrates, and extra- and intracellular parasites in the liver, myocardium, urinary bladder, and adrenal glands of the diseased animal. PCR and subsequent genotyping of parasites from fresh tissue from both cases revealed infection with T. gondii genotype II. Direct agglutination testing of blood from the healthy individual revealed high levels of T. gondii IgG antibodies. T. gondii is a potential cause of disease and lethality in captive and wild Patagonian maras, and toxoplasmosis should be considered when managing and providing veterinary care for this species.