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Differential leukocyte count method for bovine low somatic cell count milk.
H Dosogne1, F Vangroenweghe, J Mehrzad
1Ghent University, Faculty of Veterinary Medicine, Department of Physiology, Biochemistry and Biometrics, Salisburylaan 133, 9820 Merelbeke, Belgium.
Journal of Dairy Science
|April 22, 2003
Summary
A new flow cytometric method accurately quantifies leukocytes in low somatic cell count (SCC) milk. This technique differentiates lymphocytes, monocytes, neutrophils, macrophages, and apoptotic cells, aiding mastitis diagnosis in dairy cows.
Area of Science:
- Veterinary Immunology
- Dairy Science
- Flow Cytometry
Background:
- Limited methods exist for differential leukocyte counts in low somatic cell count (SCC) milk.
- Mastitis diagnosis in dairy cows often relies on milk SCC, but low SCC milk requires specialized analysis.
Purpose of the Study:
- To develop and validate a flow cytometric method for differential leukocyte counting in bovine low SCC milk.
- To characterize leukocyte populations including lymphocytes and monocytes (LM), polymorphonuclear neutrophils (PMN), mature macrophages (Mphi), and apoptotic cells.
Main Methods:
- A flow cytometric procedure involving diluted milk sample, centrifugation, leukocyte labeling with SYTO 13, and analysis.
- Optimization of processing temperature (20°C) and assessment of sample storage effects (2 days at 7°C).
Main Results:
- The method successfully identified four major leukocyte populations in low SCC milk.
- No significant impact on differential leukocyte counts was observed after 2-day storage at 7°C.
- Leukocyte profiles varied across lactation stages, with higher LM and apoptotic cells in early lactation.
Conclusions:
- A rapid, simple, accurate, and reproducible flow cytometric method for differential leukocyte counts in bovine low SCC milk was established.
- This method provides valuable insights into mammary gland health and immune status during different lactation phases.