Among the plethora of biological roles, some peptides also perform the role of an antimicrobial agent. β-defensins is one of the such cationic antimicrobial peptides with profound antimicrobial activity. Unlike the synthetic antimicrobial agents, the antimicrobial peptides are capable of killing bacteria not only in the planktonic stage but alsoat the biofilm stage. In bovines, mastitis is an intramammary infection which derives it’s redundancy for antibiotics due to the biofilm forming tendency of the causative pathogenss and subsequent prophylactic doses of which leads to antimicrobial resistance. Among the bovine β-defensins, the β-defensin 103A with high homology to human β-defensin 103 or hBD3 is abundantly expressed on epithelial membranes. Therefore, considering the β-defensin 103A as a target, we have bifurcated the present study to address the antibiofilm problem using this cationic peptide, first step involving characterizing the β-defensin 103A mature peptide coding region sequences in cattle breed (Tharparkar; Bos indicus) which is less susceptible to mastitis and secondly by using the translated peptide as a template, prediction was done insilico to screen the potential anti-biofilm activity of the peptide. In the end of this study, we have suggested the predicted peptides fragments from β-defensin 103A peptide for future studies as Immune Defense Regulators for mastitis.