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Differential leukocyte count (DSCC) in milk is considered important to improve knowledge of udder immune response. The investigations on milk DSCC were limited by the techniques available until recently, when a high-throughput tool to perform DSCC opened the way to explore these factors in rapid and economically sustainable ways. We hypothesized that DSCC alone does not fully describe the pattern of these cells, since the total amount is also influenced by milk yield and SCC. Therefore, this study was designed to describe DSCC and total amount of different leukocytes in milk during the course of lactation in cows differing in parity and in levels of SCC. This study considered 17,939 individual milk tests from 12 dairy herds in Lombardy Region, where DCC testing was applied in the period of February 2018-December 2019 (23 months). The samples were divided into two subsets-"healthy" (HS) with SCC ≤200,000 cells/mL and "inflamed" (IS) with SCC &gt;200,000 cells/mL. Cow in HS have a P + LT average between 5.0 × 10 8 and 3.0 × 10 9 cells. In IS cows, the values were 1.6 × 10 10 and 2.5 × 10 10 . Therefore, the presence of a well-defined inflammatory process increased the overall amount of polymorphonuclear neutrophils (PMN) and lymphocytes (LYM) of 1 log, from 1 × 10 9 to 1 × 10 10 . The assessment of the total amount of PMN and LYM, to our knowledge, have never been reported in scientific literature; the values observed may be proposed as benchmarks for studies on udder immune response. When data were analyzed by days in milk (DIM), they showed that cows in first and second lactation have a significantly lower amount of PMN + LYM, when compared to cows in third and higher lactation. However, these differences are numerically not very large (7%), and suggest that, in healthy animals, the number of immune cells is kept as constant as possible. In IS, the analysis of trends based on DIM showed that both DSCC and P + LT have a significant negative trend. These data suggest that only in this group, the presence of high SCC as lactation proceeds is associated with a progressive increase in the number of macrophages. To the best of our knowledge, this is the first study describing the pattern of DSCC and the total amount of PMN + LYM in relation to parity, days in milk, and SCC, and it may be considered as the first contribution in the investigation on mammary gland immune response by the means of differential cell counts in milk.

Factors Affecting the Patterns of Total Amount and Proportions of Leukocytes in Bovine Milk