Expression of CD3 and CD11b antigens on blood and mammary gland leukocytes and bacterial survival in milk of cows with experimentally induced Staphylococcus aureus mastitis

Ariel L. RivasDepartment of Biomedical Sciences, College of Veterinary Medicine, Cornell University, Ithaca, NY 14853.

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Fred W. QuimbyDepartment of Biomedical Sciences, College of Veterinary Medicine, Cornell University, Ithaca, NY 14853.

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Ozden CoksayganPopulation Medicine and Diagnostic Sciences, College of Veterinary Medicine, Cornell University, Ithaca, NY 14853.

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Argia AlbaDepartment of Microbiology, University of Navarra, Pamplona 30180, Spain.

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Ainhoa ArinaDepartment of Microbiology, University of Navarra, Pamplona 30180, Spain.

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M. Juncal ArrobasDepartment of Microbiology, University of Navarra, Pamplona 30180, Spain.

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Rubén N. GonzálezPopulation Medicine and Diagnostic Sciences, College of Veterinary Medicine, Cornell University, Ithaca, NY 14853.

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Hussni O. MohammedPopulation Medicine and Diagnostic Sciences, College of Veterinary Medicine, Cornell University, Ithaca, NY 14853.

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Donald H. LeinPopulation Medicine and Diagnostic Sciences, College of Veterinary Medicine, Cornell University, Ithaca, NY 14853.

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Abstract

Objectives—To differentiate early (1 to 8 days) from late (9 to 14 days) inflammatory phases and assess relationships between leukocyte phenotype and bacterial recovery in cows with Staphylococcus aureus-induced mastitis.

Animals—10 first-lactation Holstein cows.

Procedure—Blood and milk samples were collected from 4 or 6 cows before and after intramammary infusion of sterile broth or S aureus, respectively. Flow cytometric expression of CD3 and CD11b antigens on blood and milk leukocytes, leukocyte differential counts, bacterial counts in milk, and somatic cell counts were determined longitudinally.

Results—Density of CD3 molecules decreased on blood lymphocytes and increased on milk lymphocytes after infusion of bacteria. Density of CD11b molecules on lymphocytes and phagocytes and percentage of CD11b+ lymphocytes in milk increased significantly after infusion; maximum values were achieved during the early inflammatory phase. Density of CD3 and CD11b molecules on milk lymphocytes and macrophages, respectively, 1 day after inoculation were negatively correlated with bacterial recovery on day 1 and days 9 to 14, respectively. Density of CD11b molecules on milk macrophages and the ratios of phagocyte to lymphocyte percentages and polymorphonuclear cell to macrophage percentages in milk differentiated the early from the late inflammatory phase.

Conclusions and Clinical Relevance—Activation of bovine mammary gland macrophages and T cells in response to intramammary infusion of S aureus was associated with an inability to culture this bacterium from milk. Identification of specific inflammatory phases of S aureus-induced mastitis in cows may allow for the design of more efficacious treatment and control programs. (Am J Vet Res 2001;62:1840–1851)

Abstract

Objectives—To differentiate early (1 to 8 days) from late (9 to 14 days) inflammatory phases and assess relationships between leukocyte phenotype and bacterial recovery in cows with Staphylococcus aureus-induced mastitis.

Animals—10 first-lactation Holstein cows.

Procedure—Blood and milk samples were collected from 4 or 6 cows before and after intramammary infusion of sterile broth or S aureus, respectively. Flow cytometric expression of CD3 and CD11b antigens on blood and milk leukocytes, leukocyte differential counts, bacterial counts in milk, and somatic cell counts were determined longitudinally.

Results—Density of CD3 molecules decreased on blood lymphocytes and increased on milk lymphocytes after infusion of bacteria. Density of CD11b molecules on lymphocytes and phagocytes and percentage of CD11b+ lymphocytes in milk increased significantly after infusion; maximum values were achieved during the early inflammatory phase. Density of CD3 and CD11b molecules on milk lymphocytes and macrophages, respectively, 1 day after inoculation were negatively correlated with bacterial recovery on day 1 and days 9 to 14, respectively. Density of CD11b molecules on milk macrophages and the ratios of phagocyte to lymphocyte percentages and polymorphonuclear cell to macrophage percentages in milk differentiated the early from the late inflammatory phase.

Conclusions and Clinical Relevance—Activation of bovine mammary gland macrophages and T cells in response to intramammary infusion of S aureus was associated with an inability to culture this bacterium from milk. Identification of specific inflammatory phases of S aureus-induced mastitis in cows may allow for the design of more efficacious treatment and control programs. (Am J Vet Res 2001;62:1840–1851)

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