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There is no current information on the gene flow between mouse populations on the mountains and forest fragments found scattered throughout the Mont\u00e9r\u00e9gie region in southern Qu\u00e9bec, and whether various landscape barriers have an effect on their dispersal. We conducted a population genetics analysis on eleven<jats:italic>P. leucopus<\/jats:italic>populations using eleven microsatellite markers and showed that isolation by distance was weak, yet barriers were effective. The agricultural matrix had the least effect on gene flow, whereas highways and main rivers were effective barriers. The abundance of ticks collected from mice varied within the study area. Both ticks and mice were screened for the presence of the spirochete bacterium<jats:italic>Borrelia burgdorferi<\/jats:italic>, and we predicted areas of greater risk for Lyme disease. 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