The incidence of infectious waterborne disease in Canada continues to be a public health issue and can be associated with the source of drinking water. Millions of Canadians relying on unregulated private well water are at increased risk of disease. This study examined relationships between well and owner characteristics and the frequency of microbial testing of private wells in two southern-Ontario counties. Using multi-level logistic regression models, testing frequency (i.e., at least once per year vs. less) was modeled, as both self-reported and laboratory-validated, for associations with owner and well characteristics. For the self-reported outcome, a previous adverse test result significantly increased the odds of being classified as a frequent tester, and owners with a well-head more than 16 inches (40.6 cm) above the ground were at significantly higher odds of being classified as frequent testers compared to those with well-heads less than 16 inches above the ground and those below ground level. For the model based on the laboratory-validated outcome, the odds of an owner being a frequent tester significantly varied with the length of occupancy and the occurrence of a previous adverse result. The absence of associations between other well characteristics and testing frequency suggests that well safety education could benefit these communities.