Fully synthetic peptide vaccines possess attractive cost and safety attributes. However, peptide vaccines that induce cell-mediated immunity require both selection of appropriate peptides and the development of adjuvant formulations supporting the induction of cellular immunity. An adjuvant formulation composed of emulsigen and the synthetic CpG motif containing ODN2007 was tested in cattle for its ability to induce cellular immunity after peptide vaccination, and compared to Rv3019c DNA vaccination. Peptides from the protective Mycobacterium bovis antigen Rv3019c were included into the vaccine on the basis of their frequent and strong recognition by T cells from M. bovis infected or BCG vaccinated cattle. Following peptide vaccination, strong IFN-γ and proliferative T cell responses were observed. Proliferative, but no significant IFN-γ responses were induced by DNA vaccination. Peptide vaccination boosted responses primed by DNA vaccination. In conclusion, emulsigen and CpG motif containing ODN constitute a promising adjuvant formulation to deliver peptides to veterinary species.