4.7 Article

Bacteriophage has beneficial effects in a murine model of Klebsiella pneumoniae mastitis

Journal

JOURNAL OF DAIRY SCIENCE
Volume 104, Issue 3, Pages 3474-3484

Publisher

ELSEVIER SCIENCE INC
DOI: 10.3168/jds.2020-19094

Keywords

bovine mastitis; Klebsiella pneumoniae; bacteriophage; antibiotic alternatives; murine mastitis model

Funding

  1. Ningxia Key RD Project [2019BBF02027]
  2. Hebei Key RD Project [19226607D]
  3. National Natural Science Foundation of China [31772813, 31572587]
  4. High-end Foreign Experts Recruitment Program (Ministry of Science and Technology of the People's Republic of China, Beijing, China) [GDT20171100013]

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The bacteriophage CM8-1 isolated from dairy farm wastewater showed promising results in treating murine K. pneumoniae mastitis by reducing bacterial counts, improving mammary gland tissue morphology, and decreasing expression of pro-inflammatory factors. The phage had stable biological characteristics and effectively suppressed K. pneumoniae mastitis when administered into the mammary gland in mice 2 hours after bacterial inoculation.
Bovine mastitis caused by Klebsiella pneumoniae is usually treated with antibiotics, thereby potentially increasing antimicrobial resistance. The objective of this study was to evaluate efficacy of a bacteriophage, isolated from dairy farm wastewater, as a treatment for a murine model of K. pneumoniae mastitis. A lytic bacteriophage CM8-1 was isolated, morphological and biological characteristics were assessed with transmission electron microscopy and double-layer plate, and its genome was sequenced and analyzed. Furthermore, effectiveness of this bacteriophage for treatment of a murine model of K. pneumoniae mastitis was evaluated based on the following mammary gland characteristics: morphological changes; number of K. pneumoniae; and mRNA and protein expression of pro-inflammatory factors TNF-alpha, IL-1 beta, IL-6, and IL-8. Bacteriophage CM8-1 had an incubation period of 30 min and a burst time of 20 min. Its viability and adsorption were stable at 30 to 50 degrees C, but decreased significantly at >60 degrees C, with no significant change in viability or infectivity at pH 6 to 10. In a murine model of K. pneumoniae mastitis, injecting bacteriophage CM8-1 into the mammary gland 2 h after inoculation with K. pneumoniae resulted in reductions in bacterial counts in the murine mammary gland, improvements in mammary gland tissue morphology, and reductions in mRNA and protein expression of pro-inflammatory factors. Bacteriophage CM8-1 had stable biological characteristics and suppressed K. pneumoniae mastitis when injected into the mammary gland 2 h latera in mice bacterial inoculation.

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