Longitudinal evaluation of bovine mammary gland health status by somatic cell counting, flow cytometry, and cytology

Department of Biomedical Sciences, College of Veterinary Medicine, Cornell University, Ithaca, NY 14853, USA.

Bovine mastitis phases induced by Staphylococcus aureus were assessed in 6 lactating cows before challenge and at 1, 4-8, and 9-14 days postinoculation (dpi). Milk lymphocytes, macrophages, and polymorphonuclear cells (PMN) were counted by conventional (manual) cytology, identified by CD3+ and CD11b+ immunofluorescence and counted by flow cytometry (based on leukocyte forward and side light scatter values). Somatic cell counts (SCC) and recovery of bacteria were recorded at the same times. Preinoculation samples showed a lymphocyte-dominated composition. At 1 dpi, the percentage of PMN increased and that of lymphocytes decreased. At 4-8 dpi, PMN were predominant, but the percentage of mononuclear cells increased above that at 1 dpi and further increased by 9-14 dpi (when lymphocytes approached prechallenge values). Based on leukocyte percentages, 3 indices were created from the data: 1) the PMN/lymphocyte percentage ratio (PMN/L), 2) the PMN/macrophage percentage ratio (PMN/M), and 3) the phagocyte (PMN and macrophage)/lymphocyte percentage ratio (Phago/L). Significant correlations were found between cytologic and flow cytometric data in all of these indicators (all with P < or = 0.01). These indices identified nonmastitic, early inflammatory (1-8 dpi), and late inflammatory (9-14 dpi) animals. In contrast, SCC and bacteriology did not. Although sensitivity of the SCC was similar to that of Phago/L, the specificity of SCC was almost half that of the Phago/L index. Based on flow cytometry indicators, an algorithm for presumptive diagnosis of bovine mastitis was developed. Flow cytometry provides results as valid as those obtained by conventional (manual) cytology, shows greater ability to identify mastitic cases than does SCC, and may identify 3 mammary gland health-related conditions.