Isolation of bacteriophages specific to bovine mastitis-causing bacteria and characterization of their lytic activity in pasteurized milk
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Published:2024-01
Issue:
Volume:
Page:207-215
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ISSN:2231-0916
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Container-title:Veterinary World
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language:en
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Short-container-title:Vet World
Author:
Imklin Napakhwan1ORCID, Patikae Patinya2ORCID, Poomirut Peekarn3ORCID, Arunvipas Pipat4ORCID, Nasanit Rujikan1ORCID, Sajapitak Somchai4ORCID
Affiliation:
1. Department of Biotechnology, Faculty of Engineering and Industrial Technology, Silpakorn University, Nakhon Pathom, Thailand. 2. Veterinary Clinical Study Program, Faculty of Veterinary Medicine, Kasetsart University, Nakhon Pathom, Thailand; Faculty of Veterinary Sciences, Mahasarakham University, Maha Sarakham, Thailand. 3. Veterinary Clinical Study Program, Faculty of Veterinary Medicine, Kasetsart University, Nakhon Pathom, Thailand. 4. Department of Large Animal and Wildlife Clinical Sciences, Faculty of Veterinary Medicine, Kasetsart University, Nakhon Pathom, Thailand.
Abstract
Background and Aim: Bovine mastitis is one of the most serious issues in dairy production. It is caused by contagious and coliform pathogens such as Staphylococcus spp., Escherichia coli, and Klebsiella pneumoniae. In addition, the emergence of drug-resistant bacteria raises urgent concerns in the field of drug treatment, thus requiring the exploration of alternative treatments. Bacteriophage therapy has been shown to be a promising alternative approach for the control of antibiotic-resistant pathogens. In this study, we aimed to isolate phages specific to contagious mastitis and coliform mastitis, characterize the isolated phages, and examine their ability to lyse bacteria in pasteurized milk samples.
Materials and Methods: The Staphylococcus phage vB_Sau-RP15 isolated from raw milk in our previous study was used in this study. Other three phages, vB_Eco-RN12i1, vB_Kpn-RN14i1, and vB_Ssc-RN20i3, were isolated from wastewater using E. coli 5823, K. pneumoniae 194, and Staphylococcus sciuri MM01 as hosts, respectively. The host range and efficiency of plating (EOP) were determined following phage isolation. Moreover, the lysis activities of these phages against their hosts were investigated in pasteurized milk using a multiplicity of infections (MOIs) of 10 and 100 at 37°C. Phages were applied using individual and combination phages.
Results: According to the EOP results, all phages showed high specificity to their respective hosts. They are tailed phages with distinct morphologies. Individual phage treatments in spiked pasteurized milk with their respective bacterial hosts significantly reduced the bacterial counts in both MOI conditions during the first 2 h of the treatment (approximately 1–8 log reduction compared to the control). Although these phages specifically infected only their hosts, the phage cocktail resulted in a better result compared to the use of individual phage. However, bacterial regrowth was observed in all experiments, which may be related to the development of phage-insensitive mutants.
Conclusion: Our findings suggest that the application of phages could be used to treat bovine mastitis. Phage cocktail is suitable to promote the efficacy of phage treatment in pasteurized milk. However, when considering the use of phages in dairy cows, certain phage properties in raw milk and in vivo and ex vivo should be highlighted to ensure their effectiveness as biocontrol agents for bovine mastitis treatment.
Keywords: bovine, Escherichia coli, Klebsiella pneumoniae, mastitis, phage, Staphylococcus.
Funder
Faculty of Veterinary Medicine, Kasetsart University
Publisher
Veterinary World
Subject
General Veterinary
Reference47 articles.
1. Horpiencharoen, W., Thongratsakul, S. and Poolkhet, C. (2019) Risk factors of clinical mastitis and antimicrobial susceptibility test results of mastitis milk from dairy cattle in Western Thailand: Bayesian network analysis. Prev. Vet. Med., 164: 49–55. 2. Haxhiaj, K., Wishart, D.S. and Ametaj, B.N. (2022) Mastitis: What it is, current diagnostics, and the potential of metabolomics to identify new predictive biomarkers. Dairy, 3: 722–746. 3. Tommasoni, C., Fiore, E., Lisuzzo, A. and Gianesella, M. (2023) Mastitis in dairy cattle: On-farm diagnostics and future perspectives. Animals (Basel), 13(15): 2538. 4. Hogan, J. and Smith, K.L. (2012) Managing environmental mastitis. Vet. Clin. North Am. Food Anim. Pract., 28(2): 217–224. 5. Abebe, R., Markos, A., Abera, M. and Mekbib, B. (2023) Incidence rate, risk factors, and bacterial causes of clinical mastitis on dairy farms in Hawassa city, Southern Ethiopia. Sci. Rep., 13: 10945.
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