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We characterized the cold tolerance of natural populations of the Eastern subterranean termite (Reticulitermes flavipes (Kollar) [Isoptera, Rhinotermitidae]) in southwestern Ontario, Canada. We measured cold tolerance in workers from six colonies of termites established from Pelee Island in Lake Erie, and Point Pelee National Park. The mean critical thermal minimum, at which termites entered chill coma, ranged from 8.1 to 5.7°C. Mean supercooling points (SCP, the temperature at which individuals freeze) ranged from -4 to -4.6°C, and did not differ significantly between colonies, nor was SCP dependent on body size. Individuals survived brief exposure to low temperatures, as long as they did not freeze, but internal ice formation was always lethal, suggesting a freeze avoiding strategy. The LT50 (temperature at which 50% of individuals were killed by a 1 h exposure) was -5.1°C, but all individuals could survive -2°C for at least 72 h. Low temperature acclimation (12°C, 7 d) or hardening (4°C, 2 h) had no impact on the SCP, but acclimation did slightly increase the critical thermal minimum, making the termites less cold tolerant. We conclude that R. flavipes is not particularly cold tolerant, and likely relies on burrowing deep into the soil to avoid exposure to temperature to extremes.