A comparison was made of the supercooling points (SCPs) of questing Dermacentor variabilis adults from two populations located ca. 800 km apart on the Canadian prairies. This is also the first study to examine whether there are seasonal fluctuations in the SCP of questing D. variabilis adults. The SCPs of adult ticks from Lizard Lake Community Pasture, a recently established population in west-central Saskatchewan, varied over spring and summer, with the median SCP warming over time. In addition, the SCPs of ticks from Lizard Lake Community Pasture were significantly higher than those of adult ticks collected from Sandy Hook in Manitoba, a population that has been established for decades. The off-host survival of adults from Sandy Hook between summer and spring has been shown previously to be significantly greater than that of adults from Lizard Lake Community Pasture. The findings of the present study suggest that there may be geographical variation in the SCPs of D. variabilis adults which may be associated with differences in overwinter survival. The relatively low SCPs of questing D. variabilis adults, and the ability of some adults to survive off-host during winter, may be factors contributing to the range expansion of this tick species in Canada.