Freshwater and marine fish have been important components of human diets for millennia. The Great Lakes of North America, their tributaries and smaller regional freshwater bodies are important Native American fisheries. The ethnohistorical record, zooarchaeological remains, and isotopic values on human bone and tooth collagen indicate the importance of fish in fourteenth- through seventeenth-century ancestral Wendat diets in southern Ontario, which is bordered by three of the Great Lakes. Maize (Zea mays ssp. mays) was the primary grain of Native American agricultural systems in the centuries prior to and following sustained European presence. Here we report new Bayesian dietary mixing models using previously published δ¹³C and δ¹⁵N values on ancestral Wendat bone and tooth collagen and tooth enamel. The results confirm previous estimates from δ¹³C values that ancestral Wendat diets included high proportions of maize but indicate much higher proportions of fish than has previously been recognized. The results also suggest that terrestrial animals contributed less to ancestral Wendat diets than is typically interpreted based on zooarchaeological records.