Cattle were given Mycobacterium bovis bacillus Calmette-Guerin (BCG) in a lipid-based formulation via the oral route and tested for immune responses and protection against a challenge with virulent M. bovis. Calves were vaccinated by orally administering a pellet containing 108 colony forming units (CFU) of BCG, or 10 pellets containing a total of 109 CFU of BCG, whereas positive controls were injected subcutaneously with 106 CFU of BCG. All of the subcutaneously vaccinated calves produced positive responses in the caudal fold tuberculin skin test at 8 weeks after vaccination, whereas only 3/9 of the low dose and 6/10 of the high dose orally-vaccinated animals produced positive reactions. None of the animals produced positive reactions to the mycobacterial antigens, ESAT-6 and CFP10 in the interferon-γ (IFN-γ) test and only a total of four of the BCG-vaccinated animals produced positive responses in either the standard IFN-γ or comparative cervical skin test. Oral administration of 10 pellets of lipid-formulated BCG to cattle induced a significant level of protection against bovine tuberculosis compared to that observed in non-vaccinated animals and this level was similar to that seen in the BCG subcutaneously vaccinated animals. Oral vaccination of BCG in a lipid-formulation to calves was shown to induce some positive tuberculin skin test reactions, but could also induce protection against bovine tuberculosis.