Mycobacterium bovis (M. bovis) causes bovine tuberculosis (bTB), a biosafety and economic concern to farmers and the public. The UK has the most bTB cases in Europe, followed by Ireland and Spain. The eradication of bTB is regulated by the World Health Organisation for Animal Health (OIE), which promotes the necessity of long-term management of transmission from infected wildlife to livestock, including European badgers (Meles meles). This is essential despite the implementation of expensive testing and slaughter policies over the course of several decades. To explore host-response interactions and establish infection markers for badgers, ferrets (Mustela furo) were utilised as "clean," cost-effective laboratory-adapted mustelid surrogates. Two studies were conducted to investigate the immunopathogenesis caused by M. bovis in ferrets subjected to intratracheal (IT) inoculation or direct contact for a period of ten months. Most experimentally infected ferrets developed macroscopically and microscopically identical TB-like granulomas to those of TB-infected badgers. Differences in IFN-γ, TNF-α, IL-2, and IL-17A levels and kinetics were seen in fresh and cryopreserved peripheral mononuclear cells (PBMCs) stimulated with M. bovis antigens, PPD-B, ESAT-6, and CFP-10 post-infection. Like badgers, infected ferret PBMCs produced IFN-γ and responded similarly to PPD-B and CFP-10. The two species' IgG reactions to MPB83 were likewise comparable. Both studies revealed that IgG responses to MPB83, TNF-α, IL-17A, and IFN-γ (fresh PBMCs) responses to PPD-B correlated with total granuloma scores, while IL-2 and CXCL10 did not. Intriguingly, the in-contact ferret population showed low IFN-γ responses throughout the study. High plasma IL-17 levels, robust IgG responses to MPB83, and high IL-2 and TNF-α responses to PPD-B, ESAT-6, and CFP-10 indicate exposure to M. bovis. These responses may improve M. bovis infection detection in the field. Biomarkers for M. bovis exposure are of great benefit as they facilitate the analysis of response variability in vaccination efficacy trials.