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Characterization of bacteriophages infecting Escherichia coli associated with bovine mastitis
Published online by Cambridge University Press: 05 December 2025
Abstract
Bovine mastitis poses a significant threat to dairy production worldwide. Among the various etiologies of mastitis, Escherichia coli is a predominant environmental pathogen. Antibiotic-resistant E. coli poses substantial challenges for treating mastitis and is a threat to public health, necessitating the exploration of alternative therapeutic strategies. We studied bacteriophages as a potential alternative therapy for bovine mastitis-associated E. coli. We isolated 37 bacteriophages infecting E. coli, and characterized them for host range, growth kinetics, morphology, stability, genome fingerprinting and genome sequencing and analysis. The phages lysed between 4% and 62% of the E. coli isolates tested. Notably, 30 phages lysed bovine mastitis-associated strains. The 10 best phages selected based on host strain specificity revealed latent periods ranging from 50 to 90 min and burst sizes between 7 and 69 PFU/mL. Based on their shorter latent period and larger burst size, seven phages were subjected to transmission electron microscopy, which revealed their myovirus and siphovirus morphologies. Restriction fragment length polymorphism (RFLP) analysis of the same seven phages indicated six different patterns. The seven phages were stable at temperatures ranging from 4°C to 50°C, and at pH values ranging from 3 to 9. Whole-genome sequencing and analysis of the six phages, which showed unique RFLP patterns, predicted a lytic lifecycle, with no sequences encoding toxins or antibiotic-resistance genes. Importantly, these six phages were able to lyse multidrug-resistant and extended β-lactamase (ESBL)-producing E. coli under in vitro conditions and mastitis-associated E. coli in milk. Additionally, three phages belonging to different genera did not exhibit toxicity to mammalian cells. This study underscores the potential of bacteriophages as alternative therapeutic agents for E. coli-associated bovine mastitis. Our study has broader implications for udder and animal health, as well as the production of quality milk and dairy products, and food safety and security.
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- © The Author(s), 2025. Published by Cambridge University Press on behalf of Hannah Dairy Research Foundation.
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