Paratuberculosis (pTB), also known as Johne's disease (JD), is a contagious, chronic, and granulomatous inflammatory disease of the intestines of ruminants which is caused byMycobacterium aviumsubsp.paratuberculosis(MAP) infection, resulting in billions of dollars in economic losses worldwide. Since, currently, no effective cure is available for MAP infection, it is important to explore the genetic variants that affect the host MAP susceptibility. The aim of this study was to analyze a potential association betweenEDN2synonymous gene mutations (rs110287192,rs109651404 and rs136707411), that modifies susceptibility to pTB.EDN2 rs110287192,rs109651404andrs136707411mutations were genotyped in 68 infected and 753 healthy animals from East Anatolian Red crossbred, Anatolian Black crossbred and Holstein breed cattle by using Custom TaqMan SNP Genotyping Assays. For pTB status, serum antibody levels S/P >= 1.0 were assessed in carriers of the differentEDN2genotypes.EDN2 rs110287192mutation showed a significant association with bovine pTB (adj. p < 0.05). Forrs110287192locus, the odd ratios for GG and TG genotypes versus TT genotypes were 1.73; (95% CI = 0.34-8.59) and 0.53 (95% CI = 0.12-2.37) respectively, which indicated that proportion of TG heterozygotes were significantly higher in control animals as compared to pTB animals. On the other hand, whilers136707411mutation showed a suggestive association with pTB status in the examined cattle population (nominal p < 0.05); no association was detected betweenrs109651404genotypes and pTB status. Selecting animals againstrs110287192-GG genotype may decrease the risk of pTB in cattle of theBos taurus taurussubspecies.