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Mechanisms Involved in Milk Endogenous Proteolysis Induced by a Lipopolysaccharide Experimental Mastitis

F. Moussaoui^*, I. Michelutti, Y. Le Roux^* and F. Laurent^*

^* Laboratoire de Sciences Animales, Unité sous contrat avec l’Institut National de la Recherche Agronomique (INRA), Ecole Nationale Supérieure d’Agronomie et des Industries Alimentaires (ENSAIA), 54 505 Vandoeuvre-lès-Nancy, France
FNCL: Fédération Nationale des Coopératives Laitières 75 314 Paris Cedex 09

Corresponding author:
F. Moussaoui; e-mail:
fatima.moussaoui{at}ensaia.inpl-nancy.fr.

Experimental mastitis has been induced by the lipopolysaccharide (LPS) of Escherichia coli on six dairy cows in order to study the mechanisms involved in milk endogenous proteolysis during the inflammatory process. Variations in somatic cell count (SCC), plasmin activity, and total casein (CN) content were measured, and proteose-peptone content and the percentage of pH 4.6 insoluble peptides including -CN have been considered as indicators of endogenous proteolysis. Furthermore, polymorphonuclear neutrophils (PMN) maturity has been evaluated by optical microscopy, and proteolysis by PMN proteinases has been studied at neutral and acidic pH in order to establish a link between caseinolysis, proteinase class, and PMN maturation.

Two peaks of proteose-peptones content have been noticed for the six cows. First peak could be explained by both plasmin activity and SCC, while second peak was concomitant with a low plasmin activity but a SCC remaining high. The second peak of proteose-peptones content confirmed the role of cellular proteases in milk caseinolysis. Casein breakdown by cellular proteases was confirmed by SDS-PAGE electrophoresis, and a link between neutral proteinases activity and immature PMN recruitment was shown. Acidic proteinases activity was effective with mature PMN and during the recovery phase.

Key Words: endogenous proteolysis • lipopolysaccharide • plasmin • somatic cells

Abbreviation key: FPLC = fast-pressure liquid chromatography, LPS = lipopolysaccharide, PMN = polymorphonuclear neutrophils, PI = postinfusion, PP = proteose-peptones

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