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Outcome of bovine tuberculosis (bTB), an infectious disease caused by Mycobacterium bovis, is influenced by host genetic background. For this reason, polymorphism discovery association studies are a powerful tool for selective breeding helping disease control. Interleukin-10 (IL-10) is a regulatory cytokine produced by different cells and fine-tune immune response to bTB. Thus, in this research, we evaluated the role of the single nucleotide polymorphism g.4002C>T in IL-10 gene for susceptibility to bTB in Mediterranean water buffalo. We characterized 184 animals grouped in cases and controls and demonstrated that homozygous subjects TT are about 3 times more susceptible to bTB compared to CC homozygous. Indeed this polymorphism is responsible of amino acid substitution p. (Thr 152 Met) in the primary protein sequence which affects protein secondary structure. This polymorphism might represent a valid tool for marker assisted selection against bovine tuberculosis in water buffalo.