Recently, we found that Holstein cattle (Bos taurus taurus) displayed higher skin test prevalence and disease severity compared to zebu (Bos taurus indicus) herd-mates kept under identical husbandry conditions in Ethiopia. To determine whether these susceptibility differences were patent at the level of the innate immune system, we infected monocytes from naïve Holstein or Sahiwal zebu cattle with either live virulent Mycobacterium tuberculosis or Mycobacterium bovis. The cytokine profile following infection was compared between the two breeds by measuring IL-1α, IL-6, IL-10, IL-12 and TNF-α as well as nitric oxide in culture supernatants. Our results suggested subtle differences in the cytokine profile because only infection-induced IL-6 production was significantly increased in monocytes from the more susceptible Holstein cattle.