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Differential leukocyte count method for bovine low somatic cell count milk

(2003) JOURNAL OF DAIRY SCIENCE. 86(3). p.828-834
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Abstract
Whereas many differential leukocyte count methods for high somatic cell count (SCC) milk from mastitic cows are available, only a few have been developed for low SCC milk. We have developed a flow cytometric differential leukocyte count method for low SCC milk. The procedure consists of 1) 1.5 ml of diluted milk sample (30%, vol/vol dilution with PBS), 2) centrifugation, 3) leukocyte labeling with SYTO 13 and 4) flow cytometric analysis. Four major leukocyte populations can be clearly identified in the green fluorescence-side scatter dot plot: lymphocytes and monocytes (LM), polymorphonuclear neutrophils (PMN), mature macrophages (MO), and cells with apoptotic features based on chromatin condensation and nuclear fragmentation. The optimal processing temperature was 20degreesC. Significant differences among samples with similar differential leukocyte counts were found. Storage of milk samples during 2 d at 7degreesC had no effect on differential leukocyte count. Using the new method, differential leukocyte count was performed in low SCC milk samples from cows in early, mid, and late lactation. In accordance with previous studies, PMN and M P percentages were lower and LM percentages were higher in early lactation than in the other stages of lactation. The percentage of cells with apoptotic features was higher in early lactation than in mid and late lactation. In conclusion, a rapid, simple, accurate, and reproducible standard procedure was developed to determine the differential leukocyte count (MO, PMN, LM, and cells with apoptotic features) of bovine low SCC milk.
Keywords
POLYMORPHONUCLEAR LEUKOCYTES, DAIRY-COWS, FLOW-CYTOMETRY, APOPTOSIS, BLOOD, PHAGOCYTOSIS, NEUTROPHILS, EXPRESSION, low somatic cell count milk, ESCHERICHIA-COLI MASTITIS, flow cytometry, differential leukocyte count

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Citation

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MLA
Dosogne, Hilde et al. “Differential Leukocyte Count Method for Bovine Low Somatic Cell Count Milk.” JOURNAL OF DAIRY SCIENCE 86.3 (2003): 828–834. Print.
APA
Dosogne, H., Vangroenweghe, F., MEHRZAD, J., MASSART-LEEN, A., & Burvenich, C. (2003). Differential leukocyte count method for bovine low somatic cell count milk. JOURNAL OF DAIRY SCIENCE, 86(3), 828–834.
Chicago author-date
Dosogne, Hilde, Frédéric Vangroenweghe, J MEHRZAD, AM MASSART-LEEN, and Christian Burvenich. 2003. “Differential Leukocyte Count Method for Bovine Low Somatic Cell Count Milk.” Journal of Dairy Science 86 (3): 828–834.
Chicago author-date (all authors)
Dosogne, Hilde, Frédéric Vangroenweghe, J MEHRZAD, AM MASSART-LEEN, and Christian Burvenich. 2003. “Differential Leukocyte Count Method for Bovine Low Somatic Cell Count Milk.” Journal of Dairy Science 86 (3): 828–834.
Vancouver
1.
Dosogne H, Vangroenweghe F, MEHRZAD J, MASSART-LEEN A, Burvenich C. Differential leukocyte count method for bovine low somatic cell count milk. JOURNAL OF DAIRY SCIENCE. AMER DAIRY SCIENCE ASSOC; 2003;86(3):828–34.
IEEE
[1]
H. Dosogne, F. Vangroenweghe, J. MEHRZAD, A. MASSART-LEEN, and C. Burvenich, “Differential leukocyte count method for bovine low somatic cell count milk,” JOURNAL OF DAIRY SCIENCE, vol. 86, no. 3, pp. 828–834, 2003.
@article{348594,
  abstract     = {Whereas many differential leukocyte count methods for high somatic cell count (SCC) milk from mastitic cows are available, only a few have been developed for low SCC milk. We have developed a flow cytometric differential leukocyte count method for low SCC milk. The procedure consists of 1) 1.5 ml of diluted milk sample (30%, vol/vol dilution with PBS), 2) centrifugation, 3) leukocyte labeling with SYTO 13 and 4) flow cytometric analysis. Four major leukocyte populations can be clearly identified in the green fluorescence-side scatter dot plot: lymphocytes and monocytes (LM), polymorphonuclear neutrophils (PMN), mature macrophages (MO), and cells with apoptotic features based on chromatin condensation and nuclear fragmentation. The optimal processing temperature was 20degreesC. Significant differences among samples with similar differential leukocyte counts were found. Storage of milk samples during 2 d at 7degreesC had no effect on differential leukocyte count. Using the new method, differential leukocyte count was performed in low SCC milk samples from cows in early, mid, and late lactation. In accordance with previous studies, PMN and M P percentages were lower and LM percentages were higher in early lactation than in the other stages of lactation. The percentage of cells with apoptotic features was higher in early lactation than in mid and late lactation. In conclusion, a rapid, simple, accurate, and reproducible standard procedure was developed to determine the differential leukocyte count (MO, PMN, LM, and cells with apoptotic features) of bovine low SCC milk.},
  author       = {Dosogne, Hilde and Vangroenweghe, Frédéric and MEHRZAD, J and MASSART-LEEN, AM and Burvenich, Christian},
  issn         = {0022-0302},
  journal      = {JOURNAL OF DAIRY SCIENCE},
  keywords     = {POLYMORPHONUCLEAR LEUKOCYTES,DAIRY-COWS,FLOW-CYTOMETRY,APOPTOSIS,BLOOD,PHAGOCYTOSIS,NEUTROPHILS,EXPRESSION,low somatic cell count milk,ESCHERICHIA-COLI MASTITIS,flow cytometry,differential leukocyte count},
  language     = {eng},
  number       = {3},
  pages        = {828--834},
  publisher    = {AMER DAIRY SCIENCE ASSOC},
  title        = {Differential leukocyte count method for bovine low somatic cell count milk},
  url          = {http://jds.fass.org/cgi/reprint/86/3/828},
  volume       = {86},
  year         = {2003},
}

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