A commercially available C6 ELISA kit was used to detect antibodies induced by natural infection with Borrelia burgdorferi in dogs that lived in an area endemic for Lyme disease. Rates of infection were determined both for nonvaccinated dogs and those that had been vaccinated with a whole-cell B. burgdorferi bacterin (Lyme Vax, Fort Dodge Animal Health) before 6 months of age and were boostered annually. Vaccinated dogs had an infection rate of 5% (8 of 163), whereas 64% (25 of 39) of the non-vaccinated dogs were positive for B. burgdorferi antibodies. The preventable fraction, determined by comparing infection rates in unvaccinated and vaccinated dogs, was 92.2% (95% confidence interval: 84.3% to 96.3%). In addition, screening of nonvaccinated dogs at six Connecticut clinics (Middletown, Portland, Essex, Old Lyme, Durham, and Marlborough) with the C6 ELISA test revealed infection rates ranging from 41% to 73%, demonstrating a high level of infected dogs in the area. It was concluded that emphasis should be placed on vaccinating young dogs at risk for Lyme disease before they are exposed to infected ticks. Results of this study support the value of immunization with this whole-cell Lyme disease bacterin for dogs at risk for infection by B. burgdorferi.