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Staphylococcus aureus leucocidin, a virulence factor in bovine mastitis

Published online by Cambridge University Press:  21 March 2005

Ahmed Younis
Affiliation:
National Mastitis Reference Center, Kimron Veterinary Institute, PO Box 12, Bet-Dagan, 50250 Israel The Hebrew University of Jerusalem, Faculty of Agricultural, Food and Environmental quality Sciences, Department of Animal Sciences, PO Box 12, 76-100 Rehovot, Israel
Oleg Krifucks
Affiliation:
National Mastitis Reference Center, Kimron Veterinary Institute, PO Box 12, Bet-Dagan, 50250 Israel
Gideon Fleminger
Affiliation:
Department of Molecular Microbiology and Biotechnology, The George Wise Faculty of Life Sciences, Tel-Aviv University, Tel-Aviv 69978, Israel
Elimelech D Heller
Affiliation:
The Hebrew University of Jerusalem, Faculty of Agricultural, Food and Environmental quality Sciences, Department of Animal Sciences, PO Box 12, 76-100 Rehovot, Israel
Natan Gollop
Affiliation:
Dept. of Food Science, Agricultural Research Organization, the Volcani Center, PO Box 12, Bet-Dagan, 50250 Israel
Arthur Saran
Affiliation:
National Mastitis Reference Center, Kimron Veterinary Institute, PO Box 12, Bet-Dagan, 50250 Israel
Gabriel Leitner
Affiliation:
National Mastitis Reference Center, Kimron Veterinary Institute, PO Box 12, Bet-Dagan, 50250 Israel

Abstract

The involvement of Staphylococcus aureus exosecretions in bovine udder infection (Younis et al. 2003) suggests that four different monomer protein bands appearing between 36 and 31 kDa, are associated with the severity of the cow's infection response. Three out of these four bands have been identified by means of protein sequencing. Band B, with a MW of 35 kDa was identified as Panton-Valentaine leucocidin LukF'-PV chain- Staph. aureus; band C, with a MW of 32 kDa was identified as leucocidin chain LukM precursor- Staph. aureus; and band D was found to be similar, but not identical, to phosphatidylinositol-specific phospholipase-C-X. Bands B and C were purified by gel filtration using FPLC. The ability of these proteins to induce udder inflammation in vivo, and proliferation response in vitro and cytokine secretion were tested for both the crude exosecretions and purified bands. Three cows were inoculated intracisternally, with three quarters receiving either 0·007–0·008 mg (as total proteins) of Staph. aureus FR2449/1 bacterial exosecretion, pooled fraction 39–41 (bands B and C), or culture broth medium. The fourth quarter was left free as a control. Quarters that received fraction 39–41 of Staph. aureus FR2449/1, exhibited induced inflammation, which was indicated by increased somatic cell count and enhanced NAGase activity that was significantly higher than that of the original Staph. aureus FR2449/1 bacterial exosecretion. Proliferation tests of bovine blood lymphocytes in vitro showed that the pooled fraction 39–41 stimulated bovine proliferation of mononuclear cells much more than the original Staph. aureus FR2449/1 bacterial exosecretion. Secretion of TNF-α, IL-1β, IL-6 and IL-8 was in accordance with the contents of LukF'-PV and LukM precursor in the exosecretions. The results suggest that LukM/LukF' induce inflammation into the udder by a mechanism similar to that of LPS or by a unique mechanism(s) which requires further investigation.

Type
Research Article
Copyright
Proprietors of Journal of Dairy Research 2005

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