Bovine tuberculosis remains a major threat to the dairy industry in North America. Although the disease has nearly been eliminated in the United States and Canada, sporadic outbreaks occur with significant economic consequences to the industry. For example, >45,000 cattle were quarantined and >12,000 animals were destroyed in a recent outbreak in Western Canada that could be traced back to 6 cows that initially tested positive for bovine tuberculosis. Wildlife reservoirs, including bison and elk, that are known to transmit the disease to cattle, significantly hamper effective disease control and our ability to internationally trade live animals and meat products. Vaccines are urgently needed that not only prevent the disease in animals but also distinguish infected from vaccinated animals (DIVA vaccine). We are currently using reverse vaccinology methodology to develop a novel subunit vaccine and companion diagnostics that will meet these requirements. Reverse vaccinology is a relatively new approach to vaccine development. It is based on an unbiased screening of large numbers of potential vaccine candidates (proteins) in animal models. We have produced >80 different proteins that are currently being evaluated for their ability to protect calves from bovine tuberculosis. Promising vaccine candidates will be further characterized and optimized by formulation with adjuvants. The talk will provide an update on the status of the project.