Disease transmission between wildlife and livestock is a worldwide issue. Society needs better methods to prevent interspecies transmission to reduce disease risks. Producers have successfully used livestock protection dogs (LPDs) for thousands of years to reduce predation. We theorized that LPDs raised and bonded with cattle could be used to also reduce risk of bovine tuberculosis (Myobacterium bovis; TB) transmission between white-tailed deer (Odocoileus virginianus) and cattle by minimizing contact between the 2 species and use of cattle feed by deer. We evaluated 4 LPDs over 5 months, utilizing 2 data collection methods (direct observation and motion-activated video) on deer farms that supported higher densities than wild populations. Dogs were highly effective in preventing deer from using concentrated cattle feed (hay bales), likely the greatest risk factor of TB transmission on farms. Dogs also prevented deer from approaching cattle in core areas of pastures (near hay bales) and were very effective throughout pastures. Our research supports the theory that LPDs, specifically trained to remain with cattle, may be a practical tool to minimize potential for livestock to contract TB from infected deer in small-scale cattle operations. Where disease is present in deer, it may be possible to reduce the potential for disease transmission by employing LPDs.