Natural Bacteriocins as Potential Drug Candidates Targeting Core Proteins in Mastitis Pathogens of Dairy Cattle

Mastitis poses a major challenge in the dairy industry, with rising antibiotic-resistant strains underscoring the urgent need for alternative antimicrobial strategies. This study aimed to (i) identify essential core proteins in clinical mastitis (CM)-causing pathogens using genomic approach, and (ii) assess the efficacy of natural antimicrobial peptides as novel therapeutic agents targeting the selected core proteins for the rational management of mastitis in dairy cows. Through a core genomic analysis of 16 CM-causing pathogens, including strains of Staphylococcus aureus, S. warneri, Streptococcus agalactiae, S. uberis, Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeruginosa, P. putida, and P. asiatica, we identified 65 core proteins shared among these pathogens. Among them, ten proteins including PhoH, TrpB, FtsZ, HslV, HupB, RibH, InfA, MurA, GlxK, and Rho were found to be essential for the survival and virulence of these pathogens. Importantly, further novelty, resistance, and virulence assessments identified Rho and HupB as potential therapeutic targets. A comprehensive screening of 70 bacteriocin peptides (BPs) revealed 14 BPs that effectively interacted with both Rho and HupB proteins. Further analysis showed that BP8 and BP32 disrupt Rho protein function by blocking transcription termination process, while BP8, BP39, and BP40 prevent HupB from binding to DNA. These findings confirm the promising stability and efficacy of BP8 against both target proteins in CM-pathogens, highlighting it as a promising broad-spectrum therapeutic agent. Our computational study identified Rho and HupB as key proteins in CM-causing pathogens, which can be targeted by natural bacteriocins like BP8, suggesting its potential for developing effective and sustainable therapeutics against mastitis in dairy cattle. Author SummaryMastitis poses a significant threat to the global dairy industry, with rising antibiotic resistance necessitating alternative therapeutic strategies. This study identified essential core proteins in clinical mastitis-causing pathogens through a genomic approach and evaluated natural antimicrobial peptides (bacteriocins) as novel therapeutic agents. Through a core-genomic analysis, Rho and HupB were identified as key therapeutic targets. Bacteriocin peptides such as BP8 demonstrated promising efficacy by disrupting regular transcription termination process and DNA replication, offering a promising solution for next-generation mastitis therapies. The findings underscore the potential of BP8 as a sustainable, broad-spectrum antimicrobial agent, contributing to the rational management of mastitis in dairy cattle.