Ticks and canine sera were submitted by veterinarians from Prince Edward Island over a 15-month period spanning 3 tick seasons. The objective of the study was to determine the infection prevalence of Borrelia burgdorferi, a causative agent of Lyme disease, in the province's ticks and the seroprevalence in its dogs. It was found that 97.8% (n = 368) of ticks submitted were Ixodes scapularis, a species capable of transmitting Borrelia burgdorferi; 10.3% of these ticks [95% confidence interval (CI): 3.6% to 17.0%] were infected. Provincial canine seroprevalence for the 199 submitted samples was estimated at 3.0% (95% CI: 1.0% to 5.1%).