Relationships among quarter milk leukocyte proportions and cow and quarter-level variables under different intramammary infection statuses

The use of milk leukocyte differential (MLD) test has been proposed as a complement to somatic cell count (SCC) to assess the presence and the severity of intramammary infection. However, detailed information regarding the behavior of MLD under different physiological or pathological stages of the cow is nonexistent. The objective was to analyze the association between milk leukocyte proportions provided by a commercial automated MLD test and multiple cow and quarter-level variables. The study population consisted of 104 Holstein cows (32 primiparous and 72 multiparous) in one farm. Cows were categorized by days in milk as early (<50 DIM; n=29), middle (50–250 DIM; n=25), and late lactation (>250 DIM; n = 50). Milk from 416 quarters was collected and analyzed for lymphocytes (LYM), neutrophils (NEU), and macrophages (MAC) counts using an automated milk fluorescence microscopy system. Concurrently, a sterile composite milk sample was collected from each cow for pathogen identification through microbiological culture. Culture results were classified as no growth (NOG), gram-negative (GN), gram-positive (GP), or other (OTH). Milk leukocyte proportions varied depending on the level of total leukocyte counts (TLC; P < 0.001). Similarly, leukocyte ratios (NEU:LYM, NEU:MAC, and phagocyte:LYM) were different for multiple TLC categories (P < 0.05). There was no association between parity number and MLD; however, cows in early lactation had the greatest proportions of NEU and LYM. Leukocyte ratios varied depending on parity number and stage of lactation. Cows in the medium milk-yield category had the smallest proportions of NEU and LYM, and there was significant variation in leukocyte ratios, depending on the level of milk yield. In healthy quarters, MLD were not associated with quarter position; however, the NEU:MAC ratio was greater in rear quarters than in front quarters. In quarters with TLC >100,000, NEU% was greater in rear quarters than in front quarters (P = 0.03). For quarters with pathogen growth, TLC was greatest for GN followed by OTH and GP (P < 0.001). Milk LD depended on the isolated pathogen group, although the magnitudes of the differences were small. Although the changes in the proportions of leukocytes in milk were associated with categories of TLC, levels of milk yield, and mastitis-causing pathogen groups, the deviations were small in magnitude. Additional research is necessary to determine the potential applications for this methodology.