title: Differential somatic cell count-A novel method for routine mastitis screening in the frame of Dairy Herd Improvement testing programs.
abstract: Somatic cell count (SCC) is used as key indicator in mastitis screening programs typically applied in the frame of Dairy Herd Improvement (DHI) testing programs. However, mastitis is still causing tremendous economic losses to the dairy industry. Hence, new biomarkers are needed that can be used for enhanced management of mastitis on dairy farms. Besides the determination of SCC, differentiation of cells has been described to be beneficial for a more definite description of the actual udder health status of dairy cows. The aim of this study was to develop a new method for rapid and simultaneous determination of SCC and a new parameter, differential somatic cell count (DSCC), in individual cow milk samples using flow cytometry. The method is sought to be applied in central milk testing laboratories, so that existing DHI infrastructures can be used. The DSCC represents the combined proportion of polymorphonuclear leukocytes (PMN) and lymphocytes expressed in percentage. The proportion of macrophages can be calculated by subtracting DSCC from 100%. Our research revealed increasing proportions of PMN, but decreasing proportions of macrophages as SCC increased. However, lymphocytes occurred fairly constantly with low proportions across the entire SCC range. Hence, the DSCC parameter reflects the antidromic trend of PMN and macrophages. Fluorescence microscopy was used to evaluate the specificity of the new Foss DSCC method in terms of DSCC and a high correlation was found. Apart from that, the accuracy of cell differentiation using the Foss DSCC method was confirmed in a cell sorting trial. Total SCC could be determined equally well using the new method as compared with existing methods. The new method was further proven to be robust toward a range of method and milk-sample-related factors. In an initial field trial, regular DHI samples of a local dairy herd were analyzed. The DSCC values occurred in a broad range from 34 to 79% in samples with <400,000 cells/mL. Higher DSCC values (53-89%) were found in samples with >400,000 cells/mL. In conclusion, the new Foss DSCC method allows reliable, repeatable, fast, robust, and accurate determination of both DSCC and SCC at low cost. This, in turn, provides more accurate information on the actual udder health status of dairy cows. The practical application of DSCC in the frame of DHI testing programs, however, needs further investigation.
authors.affiliation: Foss Analytical A/S, Foss Allé 1, 3400 Hillerød, Denmark.
authors.forename: Malin
authors.initials: M
authors.lastname: Damm
authors.affiliation: Foss Analytical A/S, Foss Allé 1, 3400 Hillerød, Denmark.
authors.forename: Claus
authors.initials: C
authors.lastname: Holm
authors.affiliation: Foss Analytical A/S, Foss Allé 1, 3400 Hillerød, Denmark.
authors.forename: Mette
authors.initials: M
authors.lastname: Blaabjerg
authors.affiliation: Foss Analytical A/S, Foss Allé 1, 3400 Hillerød, Denmark.
authors.forename: Morten Novak
authors.initials: MN
authors.lastname: Bro
authors.affiliation: Foss Analytical A/S, Foss Allé 1, 3400 Hillerød, Denmark. Electronic address: das@foss.dk.
authors.forename: Daniel
authors.initials: D
authors.lastname: Schwarz
mesh_terms: D000818:Animals; D002417:Cattle; D002452:Cell Count / Q000379:methods / Q000662:veterinary; D005260:Female; D005434:Flow Cytometry / Q000662:veterinary; D008214:Lymphocytes / Q000166:cytology; D008264:Macrophages / Q000166:cytology; D008321:Mammary Glands, Animal / Q000473:pathology; D008414:Mastitis, Bovine / Q000473:pathology*; D008892:Milk / Q000166:cytology*; D009504:Neutrophils / Q000166:cytology
keywords: differential somatic cell count; mastitis; somatic cell count; udder health