Yuxiang Shi1,2, Wenpeng Zhao1, Gang Liu1, Tariq Ali1,3, Peng Chen1, Yongxia Liu4, John P Kastelic5, Bo Han6, Jian Gao7. 1. College of Veterinary Medicine, China Agricultural University, Yuan Ming Yuan West Road No. 2, Haidian District, 100193, Beijing, P. R. China. 2. College of Life Sciences and Food Engineering, Hebei University of Engineering, 056038, Handan, Hebei, P.R. China. 3. Center of Microbiology & Biotechnology, Veterinary Research Institute, Peshawar, Pakistan. 4. College of Veterinary Medicine, Shandong Agricultural University, 271018, Taìan, China. 5. Department of Production Animal Health, Faculty of Veterinary Medicine, University of Calgary, T2N 4N1, Calgary, AB, Canada. 6. College of Veterinary Medicine, China Agricultural University, Yuan Ming Yuan West Road No. 2, Haidian District, 100193, Beijing, P. R. China. hanbo@cau.edu.cn. 7. College of Veterinary Medicine, China Agricultural University, Yuan Ming Yuan West Road No. 2, Haidian District, 100193, Beijing, P. R. China. gaojian2016@cau.edu.cn.
Abstract
BACKGROUND: Klebsiella pneumoniae, an environmental pathogen causing mastitis in dairy cattle, is often resistant to antibiotics. K. pneumoniae was used as the host bacteria to support bacteriophage replication; 2 bacteriophages, CM8-1 and SJT-2 were isolated and considered to have therapeutic potential. In the present study, we determined the ability of these 2 bacteriophages to mitigate cytotoxicity, pathomorphological changes, inflammatory responses and apoptosis induced by K. pneumoniae (bacteriophage to K. pneumoniae MOI 1:10) in bovine mammary epithelial cells (bMECs) cultured in vitro. RESULTS: Bacteriophages reduced bacterial adhesion and invasion and cytotoxicity (lactate dehydrogenase release). Morphological changes in bMECs, including swelling, shrinkage, necrosis and hematoxylin and eosin staining of cytoplasm, were apparent 4 to 8 h after infection with K. pneumoniae, but each bacteriophage significantly suppressed damage and decreased TNF-α and IL-1β concentrations. K. pneumoniae enhanced mRNA expression of TLR4, NF-κB, TNF-α, IL-1β, IL-6, IL-8, caspase-3, caspase-9 and cyt-c in bMECs and increased apoptosis of bMECs, although these effects were mitigated by treatment with either bacteriophage for 8 h. CONCLUSIONS: Bacteriophages CM8-1 and SJT-2 mitigated K. pneumoniae-induced inflammation in bMECs cultured in vitro. Therefore, the potential of these bacteriophages for treating mastitis in cows should be determined in clinical trials.
BACKGROUND:Klebsiella pneumoniae, an environmental pathogen causing mastitis in dairy cattle, is often resistant to antibiotics. K. pneumoniae was used as the host bacteria to support bacteriophage replication; 2 bacteriophages, CM8-1 and SJT-2 were isolated and considered to have therapeutic potential. In the present study, we determined the ability of these 2 bacteriophages to mitigate cytotoxicity, pathomorphological changes, inflammatory responses and apoptosis induced by K. pneumoniae (bacteriophage to K. pneumoniae MOI 1:10) in bovine mammary epithelial cells (bMECs) cultured in vitro. RESULTS: Bacteriophages reduced bacterial adhesion and invasion and cytotoxicity (lactate dehydrogenase release). Morphological changes in bMECs, including swelling, shrinkage, necrosis and hematoxylin and eosin staining of cytoplasm, were apparent 4 to 8 h after infection with K. pneumoniae, but each bacteriophage significantly suppressed damage and decreased TNF-α and IL-1β concentrations. K. pneumoniae enhanced mRNA expression of TLR4, NF-κB, TNF-α, IL-1β, IL-6, IL-8, caspase-3, caspase-9 and cyt-c in bMECs and increased apoptosis of bMECs, although these effects were mitigated by treatment with either bacteriophage for 8 h. CONCLUSIONS: Bacteriophages CM8-1 and SJT-2 mitigated K. pneumoniae-induced inflammation in bMECs cultured in vitro. Therefore, the potential of these bacteriophages for treating mastitis in cows should be determined in clinical trials.
Authors: Jia Cheng; Jv Zhang; Bo Han; Herman W Barkema; Eduardo R Cobo; John P Kastelic; Man Zhou; Yuxiang Shi; Jianfang Wang; Rui Yang; Jian Gao Journal: J Dairy Sci Date: 2020-02-07 Impact factor: 4.034
Authors: Jia Cheng; Weijie Qu; Herman W Barkema; Diego B Nobrega; Jian Gao; Gang Liu; Jeroen De Buck; John P Kastelic; Hong Sun; Bo Han Journal: J Dairy Sci Date: 2019-01-11 Impact factor: 4.034