Document Detail

Differential leukocyte count method for bovine low somatic cell count milk.
MedLine Citation:
PMID:  12703619     Owner:  NLM     Status:  MEDLINE    
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 (Mphi), and cells with apoptotic features based on chromatin condensation and nuclear fragmentation. The optimal processing temperature was 20 degrees C. Significant differences among samples with similar differential leukocyte counts were found. Storage of milk samples during 2 d at 7 degrees C 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 Mphi 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 (Mphi, PMN, LM, and cells with apoptotic features) of bovine low SCC milk.
H Dosogne; F Vangroenweghe; J Mehrzad; A M Massart-Leën; C Burvenich
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Publication Detail:
Type:  Comparative Study; Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Journal of dairy science     Volume:  86     ISSN:  0022-0302     ISO Abbreviation:  J. Dairy Sci.     Publication Date:  2003 Mar 
Date Detail:
Created Date:  2003-04-21     Completed Date:  2003-05-27     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  2985126R     Medline TA:  J Dairy Sci     Country:  United States    
Other Details:
Languages:  eng     Pagination:  828-34     Citation Subset:  IM    
Ghent University, Faculty of Veterinary Medicine, Department of Physiology, Biochemistry and Biometrics, Salisburylaan 133, 9820 Merelbeke, Belgium.
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MeSH Terms
Cell Count*
Flow Cytometry
Leukocyte Count / methods*
Mastitis, Bovine / pathology
Microscopy, Confocal
Milk / cytology*

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