Longitudinal Evaluation of Bovine Mammary Gland Health Status by Somatic Cell Counting, Flow Cytometry, and Cytology

Bovine mastitis phases induced by Staphylococcus aureus were assessed in 6 lactating cows before challenge and at 1, 4–8, and 9–14 days postinoculation (dpi). Milk lymphocytes, macrophages, and polymorphonuclear cells (PMN) were counted by conventional (manual) cytology, identified by CD3+ and CD11b+ immunofluorescence and counted by flow cytometry (based on leukocyte forward and side light scatter values). Somatic cell counts (SCC) and recovery of bacteria were recorded at the same times. Preinoculation samples showed a lymphocyte-dominated composition. At 1 dpi, the percentage of PMN increased and that of lymphocytes decreased. At 4–8 dpi, PMN were predominant, but the percentage of mononuclear cells increased above that at 1 dpi and further increased by 9–14 dpi (when lymphocytes approached prechallenge values). Based on leukocyte percentages, 3 indices were created from the data: 1) the PMN/lymphocyte percentage ratio (PMN/L), 2) the PMN/macrophage percentage ratio (PMN/M), and 3) the phagocyte (PMN and macrophage)/lymphocyte percentage ratio (Phago/L). Significant correlations were found between cytologic and flow cytometric data in all of these indicators (all with P ≤ 0.01). These indices identified nonmastitic, early inflammatory (1–8 dpi), and late inflammatory (9–14 dpi) animals. In contrast, SCC and bacteriology did not. Although sensitivity of the SCC was similar to that of Phago/L, the specificity of SCC was almost half that of the Phago/L index. Based on flow cytometry indicators, an algorithm for presumptive diagnosis of bovine mastitis was developed. Flow cytometry provides results as valid as those obtained by conventional (manual) cytology, shows greater ability to identify mastitic cases than does SCC, and may identify 3 mammary gland health-related conditions.

[1]  M. Franz,et al.  Initiating the inflammatory phase of incisional healing prior to tissue injury. , 2000, The Journal of surgical research.

[2]  H. Volk,et al.  Immunomodulatory therapies in sepsis , 2000, Intensive Care Medicine.

[3]  M. Quinn,et al.  Selective Recruitment of T-Cell Subsets to the Udder during Staphylococcal and Streptococcal Mastitis: Analysis of Lymphocyte Subsets and Adhesion Molecule Expression , 1999, Infection and Immunity.

[4]  P. Moroni,et al.  Study on the relationship between milk immune factors and Staphylococcus aureus intramammary infections in dairy cows , 1999, Journal of Dairy Research.

[5]  V. Broudy,et al.  Deletion of the extracellular membrane-distal cytokine receptor homology module of Mpl results in constitutive cell growth and loss of thrombopoietin binding. , 1999, Blood.

[6]  Q. Hamid,et al.  Immunohistochemical characterization of inflammatory and proliferative events during chronic rejection in rat lung allografts. , 1997, Transplantation.

[7]  D. Wilson,et al.  Association between management practices, dairy herd characteristics, and somatic cell count of bulk tank milk. , 1997, Journal of the American Veterinary Medical Association.

[8]  T. J. Yang,et al.  Lactation Stage‐Dependent Changes of Lymphocyte Subpopulations in Mammary Secretions: Inversion of CD4+/CD8+ T Cell Ratios at Parturition , 1997, American journal of reproductive immunology.

[9]  C. Batt,et al.  Diversity of Streptococcus agalactiae and Staphylococcus aureus ribotypes recovered from New York dairy herds. , 1997, American journal of veterinary research.

[10]  S. Goto,et al.  Characterization of cytokine and iNOS mRNA expression in situ during the course of experimental autoimmune myocarditis in rats. , 1997, Journal of molecular and cellular cardiology.

[11]  C. Mackay,et al.  Characterisation of leucocytic somatic cells in bovine milk. , 1996, Research in veterinary science.

[12]  L. Sordillo,et al.  Diminished Mammary Gland Lymphocyte Functions Parallel Shifts in Trafficking Patterns during the Postpartum Period , 1996, Proceedings of the Society for Experimental Biology and Medicine. Society for Experimental Biology and Medicine.

[13]  A. Pockley,et al.  Flow cytometric analysis of peripheral blood lymphocyte subset light scatter characteristics as a means of monitoring the development of rat small bowel allograft rejection , 1995, Clinical and experimental immunology.

[14]  Y. Le Roux,et al.  Proteolysis in samples of quarter milk with varying somatic cell counts. 1. Comparison of some indicators of endogenous proteolysis in milk. , 1995, Journal of dairy science.

[15]  D. Stites,et al.  Detection and quantification of live, apoptotic, and necrotic human peripheral lymphocytes by single-laser flow cytometry , 1995, Clinical and diagnostic laboratory immunology.

[16]  P. Bayer,et al.  Precision and accuracy of monocyte counting. Comparison of two hematology analyzers, the manual differential and flow cytometry. , 1995, American Journal of Clinical Pathology.

[17]  T. Springer,et al.  Traffic signals on endothelium for lymphocyte recirculation and leukocyte emigration. , 1995, Annual review of physiology.

[18]  J. Cullor,et al.  Bovine milk lymphocytes display the phenotype of memory T cells and are predominantly CD8+. , 1994, Cellular immunology.

[19]  M. Paape,et al.  Role of the neutrophil leucocyte in the local and systemic reactions during experimentally induced E. coli mastitis in cows immediately after calving. , 1994, The Veterinary quarterly.

[20]  B. Mallard,et al.  Genetic impact on the risk of intramammary infection following Staphylococcus aureus challenge. , 1994, Journal of dairy science.

[21]  M. Paape,et al.  Flow cytometric analysis of neutrophils in cows' milk. , 1993, American journal of veterinary research.

[22]  L. Fox,et al.  Suppression of proliferative response of BoCD4+ T lymphocytes by activated BoCD8+ T lymphocytes in the mammary gland of cows with Staphylococcus aureus mastitis. , 1993, Veterinary immunology and immunopathology.

[23]  K. Ostensson.  Total and differential leukocyte counts, N-acetyl-beta-D-glucosaminidase activity, and serum albumin content in foremilk and residual milk during endotoxin-induced mastitis in cows. , 1993, American journal of veterinary research.

[24]  L. Fox,et al.  Bovine mononuclear leukocyte subpopulations in peripheral blood and mammary gland secretions during lactation. , 1992, Journal of dairy science.

[25]  J. Fuhr,et al.  Flow cytometric analysis of pulmonary fluids and cells for the detection of malignancies. , 1992, The American journal of pathology.

[26]  K. Ostensson,et al.  Flow cytofluorometric studies on the alteration of leukocyte populations in blood and milk during endotoxin-induced mastitis in cows. , 1990, American journal of veterinary research.

[27]  D. Garner,et al.  Identification of inflammatory cells in bovine milk by flow cytometry. , 1988, Cytometry.

[28]  G. Åström,et al.  Cells found in non-infected and staphylococcus-infected bovine mammary quarters and their ability to phagocytose fluorescent microspheres. , 1986, Zentralblatt fur Veterinarmedizin. Reihe B. Journal of veterinary medicine. Series B.

[29]  R. Miller,et al.  Relationships of Milk Somatic Cell Counts to Daily Milk Yield and Composition , 1983 .

[30]  S. C. Nickerson,et al.  Cells in local reaction to experimental Staphylococcus aureus infection in bovine mammary gland. , 1982, Journal of dairy science.

[31]  B J Kitchen,et al.  Bovine mastitis: milk compositional changes and related diagnostic tests , 1981, Journal of Dairy Research.

[32]  L. Herzenberg,et al.  ANALYSIS OF CELL POPULATIONS WITH A FLUORESCENCE‐ACTIVATED CELL SORTER * , 1975, Annals of the New York Academy of Sciences.