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PLoS ONE

Blum, S.E., National Mastitis Reference Center, Department of Bacteriology, Kimron Veterinary Institute, Bet Dagan, Israel; Krifucks, O., National Mastitis Reference Center, Department of Bacteriology, Kimron Veterinary Institute, Bet Dagan, Israel; Lavon, Y., Israel Cattle Breeders Association, Caesarea, Israel; Leitner, G., National Mastitis Reference Center, Department of Bacteriology, Kimron Veterinary Institute, Bet Dagan, Israel

Recent findings have indicated that secreted phospholipases A2 (sPLA2s) have anti-inflammatory functions, including relief of symptoms in a mouse model of mastitis. This prompted us to investigate the therapeutic application of sPLA2, PLA2G1B, for bovine mastitis. Initial testing of PLA2G1B’s effect on bovine mammary epithelial cell (bMEC) line PS revealed no changes in cell viability or cytokine-secretion pattern. However, when cells were first treated with lipopolysaccharide endotoxin (LPS) or live bacteria (Escherichia coli or Staphylococcus aureus), incubation with PLA2G1B significantly improved cell viability, suggesting involvement of sPLA2s in protecting membranes from lipid-peroxidation damage, rather than a bactericidal action. When PLA2G1B was applied simultaneously with LPS, a significant short-term reduction in interleukin-8 secretion was observed compared with bMECs treated only with LPS, supporting previous reports that PLA2G1B affects interleukin-8 signaling in similar cells. Following the favorable outcome of the in vitro experiments, we tested PLA2G1B in vivo by mammary infusion into infected glands. In one of a small sample (n = 4) of lactating cows chronically infected with Streptococcus dysgalactiae, a single PLA2G1B treatment completely cleared inflammation and bacteria, demonstrating its potential to cure subclinical mastitis. PLA2G1B treatment did not affect coagulase-negative staphylococci infection. These types of mastitis may involve formation of a resistant biofilm, and its elimination may relate to sPLA2s’ characteristic ability to aggregate with cellular debris, facilitating their internalization by macrophages. In a bovine model of clinical mastitis based on introduction of E. coli via the streak canal, a single mammary infusion of PLA2G1B led to faster recovery to pre-infection milk-yield levels and decrease of somatic cell counts. In this case, all of sPLA2s’ modes of resolving inflammation may apply, including competitive binding of the sPLA2s’ receptor, the inactivation of which confers resistance to endotoxic shock. Hence, this study strongly supports further research into PLA2G1B as a cure for bovine mastitis. © 2018 Seroussi et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

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Application of pancreatic phospholipase A2 for treatment of bovine mastitis
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Blum, S.E., National Mastitis Reference Center, Department of Bacteriology, Kimron Veterinary Institute, Bet Dagan, Israel; Krifucks, O., National Mastitis Reference Center, Department of Bacteriology, Kimron Veterinary Institute, Bet Dagan, Israel; Lavon, Y., Israel Cattle Breeders Association, Caesarea, Israel; Leitner, G., National Mastitis Reference Center, Department of Bacteriology, Kimron Veterinary Institute, Bet Dagan, Israel

Application of pancreatic phospholipase A2 for treatment of bovine mastitis .

Recent findings have indicated that secreted phospholipases A2 (sPLA2s) have anti-inflammatory functions, including relief of symptoms in a mouse model of mastitis. This prompted us to investigate the therapeutic application of sPLA2, PLA2G1B, for bovine mastitis. Initial testing of PLA2G1B’s effect on bovine mammary epithelial cell (bMEC) line PS revealed no changes in cell viability or cytokine-secretion pattern. However, when cells were first treated with lipopolysaccharide endotoxin (LPS) or live bacteria (Escherichia coli or Staphylococcus aureus), incubation with PLA2G1B significantly improved cell viability, suggesting involvement of sPLA2s in protecting membranes from lipid-peroxidation damage, rather than a bactericidal action. When PLA2G1B was applied simultaneously with LPS, a significant short-term reduction in interleukin-8 secretion was observed compared with bMECs treated only with LPS, supporting previous reports that PLA2G1B affects interleukin-8 signaling in similar cells. Following the favorable outcome of the in vitro experiments, we tested PLA2G1B in vivo by mammary infusion into infected glands. In one of a small sample (n = 4) of lactating cows chronically infected with Streptococcus dysgalactiae, a single PLA2G1B treatment completely cleared inflammation and bacteria, demonstrating its potential to cure subclinical mastitis. PLA2G1B treatment did not affect coagulase-negative staphylococci infection. These types of mastitis may involve formation of a resistant biofilm, and its elimination may relate to sPLA2s’ characteristic ability to aggregate with cellular debris, facilitating their internalization by macrophages. In a bovine model of clinical mastitis based on introduction of E. coli via the streak canal, a single mammary infusion of PLA2G1B led to faster recovery to pre-infection milk-yield levels and decrease of somatic cell counts. In this case, all of sPLA2s’ modes of resolving inflammation may apply, including competitive binding of the sPLA2s’ receptor, the inactivation of which confers resistance to endotoxic shock. Hence, this study strongly supports further research into PLA2G1B as a cure for bovine mastitis. © 2018 Seroussi et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

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