Reticulitermes flavipes (Kollar) is best known throughout southwestern counties of Ontario, Canada, as an urban pest. Little is known, however, of the presence of this termite in nonurban settings in Ontario. In this study, we confirmed the existence of a population of R. flavipes on the shores of Lake Erie within Point Pelee National Park. A systematic trap survey conducted along the park's semi-vegetated west beach revealed several zones of termite activity. An analysis of trap response at one location indicated an association between termite activity and smaller sized, moderately decayed woody debris. Further, microsatellite DNA analysis suggested the presence of at least three genetically distinct colonies, each likely headed by multiple inbred reproductives. Together these data suggest that termite activity is linked to food quality, and that single colonies are potentially long-lived through multiple generations of inbreeding. Assuming it is derived from a population reported from Point Pelee in 1929, the study population is the oldest known eastern subterranean termite population in Ontario.