Due to the inadequate inheritance of resistance to digital dermatitis (DD), breeding for it is difficult. There may be a connection between genetic and metabolic variables and the development of DD. In this study, 80 pure Italian buffaloes were utilized to examine the relationship between immunological and metabolic gene mutations and digital dermatitis (DD). The incidence and history of digital dermatitis were screened, and the animals were divided into two groups: the affected group (n = 40), and the resistant (healthy) group (n = 40). Ten millilitres of blood were collected from each buffalo for DNA extraction. PCR-DNA sequencing of immune (TLR4, IL-8, CATH, IL-17, IL-13, DEFB123, ASB16, and SCART1) and metabolic (CMPK2, POPDC3, KIF26A, SYT8, ASPG, NRXN2, RND1, GPHN, LPCAT1, FPGT, TNN13K, and NDUFS6) genes revealed coding single nucleotide polymorphisms (SNPs) between DD healthy and affected buffaloes. A substantial variation was observed in the frequency of genes, based on Fisher's exact test of discovered SNPs. The identified SNPs could be employed in marker-assisted selection for digital dermatitis resistance/ susceptibility in buffaloes. These findings propose a hopeful approach for limiting digital dermatitis in buffalo through discriminating breeding of animals established on genetic markers accompanying genetic resistance to infection.