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Original Research Article The objective of study is to investigate both the phylogenetic and identification of highly conserved domains of Clostridium perfringens genes (cpa, cpb, etx and iap) isolated from dairy cattle clinically infected with mastitis using bioinformatics approach to explore more information about (cpa, cpb, etx and iap) proteins and applying insilico analysis for designing potential B-epitope subunit vaccine against C. Perfringens type A in dairy cattle farms. Five isolates were confirmed to be C. Perfringens type A in a prevalence of 7.14% (5/70). The multiplex PCR revealed the presence of the CPA gene (α toxin) of a approximate size of 402 bp while the CPB gene (B toxin), ETX gene (ε toxin) and IAP gene (iota toxin) went undetected. Two conserved domains regions in the nucleotide sequence of C. Perfringens type A (CPA) protein were detected Zn_dep_PLPC (Zinc dependent phospholipase C-alpha toxin) (63356 bp) and Zn_dep_PLPC (Zinc dependent phospholipase C-alpha toxin) (69-356 bp). Phylogenetic analysis of CPA gene showed a high identity (99-99.5%) for C. perfringens type A ASM strain with C. perfringens type A strains present in GeneBank. The insilico study was used to identify peptide fragments from Alpha toxin of C. perfringens type A that can be efficiently used for the designing and development of B-epitope based vaccine. B-cell epitopes are predicted using integrated computational tools. IEDB server was used to predict B-cell epitopes on the basis of different essential parameters like antigenicity, allergenicity, surface accessibility and flexibility. Based on the results interpretation, the top peptide sequences were (SQKGTAG, NSQKGTA, QKGTAGY, KGTAGYI) obtained as potential B-cell epitopes.

Phylogenetic and Gene Sequence Analysis of Clostridium Perfringens Genes (Cpa, Cpb, Etx and Iap) Isolated from Clinical Mastitis in Cattle Dairy Farms