Globally, ticks are becoming more problematic as agricultural pests and vectors of tick-borne diseases (TBDs), many of which are shared between animals and people. Veterinary professionals, including veterinarians and non-veterinarians, are considered a vulnerable group through occupational exposure. To inform educational interventions at the intrapersonal level, a common approach is to first assess knowledge, attitudes, and practices (KAP) of target populations. Thus, our objective was to assess KAP of veterinary professionals in Ohio, a state faced with expanding and invading ticks of medical and veterinary concern. Using a convenience sample, we surveyed 178 veterinary professionals across Ohio with an electronic questionnaire that covered knowledge, attitudes, practices, exposures, demography, education, and surveillance regarding ticks and TBDs. We found that veterinary professionals had cautionary attitudes towards ticks and TBDs and practiced prevention for themselves and their patients, even when tick exposures were infrequently reported. However, veterinary professionals significantly lacked in knowledge regarding tick biology and local TBD epidemiology. Furthermore, we found that knowledge regarding tick biology and attitudes towards ticks and TBDs did not associate with practices. Instead, we found that veterinarian status and routine tick checks of patients were associated with frequent discussions of tick prevention with clients. Our findings emphasize that most tick exposures of veterinary professionals are occupational, and thus prevention should start at the workplace. Cultivating knowledge of veterinary professionals regarding tick biology and local TBD epidemiology may lead to greater motivation and confidence with identifying ticks and testing for TBDs, which may also increase the diagnostic capacity of veterinary professionals for tick and TBD surveillance. Because veterinary professionals typically interact with both animals and their owners, improving their KAP regarding ticks and TBDs, can in turn promote animal, human, and environmental health within a “One Health” context.