Differential somatic cell count—A novel method for routine mastitis screening in the frame of Dairy Herd Improvement testing programs Malin Damm, Claus.

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Differential somatic cell count—A novel method for routine mastitis screening in the frame of Dairy Herd Improvement testing programs Malin Damm, Claus Holm, Mette Blaabjerg, Morten Novak Bro, Daniel Schwarz Journal of Dairy Science Volume 100, Issue 6, Pages 4926-4940 (June 2017) DOI: 10.3168/jds.2016-12409 Copyright © 2017 American Dairy Science Association Terms and Conditions

Figure 1 Proportions of (a) lymphocytes, (b) macrophages, and (c) PMN determined by the Foss differential somatic cell count (DSCC) method applied on a fluorescence microscope depending on SCC. In total, 293 routinely available cow-composite and DHI samples were analyzed. Data are illustrated in combination with logarithmic trendlines. Each symbol represents the result of one cow-composite sample, but overlapping is possible. Journal of Dairy Science 2017 100, 4926-4940DOI: (10.3168/jds.2016-12409) Copyright © 2017 American Dairy Science Association Terms and Conditions

Figure 2 Schematic diagram of the counting principle. (A) Separation between background (gray dots) and milk cells (black dots) fluorescence information and determination of SCC in cells per milliliter. (B) Only somatic cells from A are used for determination of differential somatic cell count (DSCC) by differentiating macrophages (light gray dots) from lymphocytes and PMN (dark gray dots). FL = fluorescence channel. Journal of Dairy Science 2017 100, 4926-4940DOI: (10.3168/jds.2016-12409) Copyright © 2017 American Dairy Science Association Terms and Conditions

Figure 3 Somatic cell count results of n = 996 routinely available cow-composite and DHI samples analyzed on the Fossomatic FC (Foss Analytical A/S, Hillerød, Denmark) and, with the new Foss differential somatic cell count (DSCC) method, on a BD LSR II instrument (Becton, Dickinson and Company, Franklin Lakes, NJ). Journal of Dairy Science 2017 100, 4926-4940DOI: (10.3168/jds.2016-12409) Copyright © 2017 American Dairy Science Association Terms and Conditions

Figure 4 Specificity of the Foss differential somatic cell count (DSCC) method shown as correlation between differential somatic cell count (DSCC) percentage obtained with a BD Accuri C6 cytometer (Becton, Dickinson and Company, Franklin Lakes, NJ) and with fluorescence microscopy (R2 = 0.7). Data are shown for 180 routinely available cow-composite samples, with SCC between 25,000 and 2,500,000 cells/mL. Journal of Dairy Science 2017 100, 4926-4940DOI: (10.3168/jds.2016-12409) Copyright © 2017 American Dairy Science Association Terms and Conditions

Figure 5 Representative examples of macrophage (A), PMN (B), and lymphocyte (C) are shown (40-fold magnification) analyzed using fluorescence microscopy. Color version available online. Journal of Dairy Science 2017 100, 4926-4940DOI: (10.3168/jds.2016-12409) Copyright © 2017 American Dairy Science Association Terms and Conditions

Figure 6 Results of the investigation of the relation between different methods for cell differentiation: (1) Foss differential somatic cell count (DSCC) method, (2) fluorescence microscopy, (3) flow cytometry after antibody-staining, and (4) light microscopy. Data are illustrated in combination with linear trendlines. Each symbol represents the result of one cow-composite sample, but overlapping is possible. Journal of Dairy Science 2017 100, 4926-4940DOI: (10.3168/jds.2016-12409) Copyright © 2017 American Dairy Science Association Terms and Conditions

Figure 7 Differential somatic cell count (DSCC) versus SCC plot for results of initial field study using DHI samples from one dairy farm (n = 188). Data are illustrated in combination with a linear trendline. Each symbol represents the result of one cow-composite sample, but overlapping is possible. Journal of Dairy Science 2017 100, 4926-4940DOI: (10.3168/jds.2016-12409) Copyright © 2017 American Dairy Science Association Terms and Conditions