Fig. 5

Raw IL-1β concentration vs. IL-1β concentration standardised to LPS. Bovine mammary macrophages (BMMOs) were isolated from milk and seeded into culture dishes at ~ 50,000 BMMOs/well. BMMOs were challenged with heat-killed S. uberis strain 0140J at a multiplicity of infection (MOI) of 50:1 0140J:BMMO. Supernatants were collected 20 h after challenge and the concentration of IL-1β was measured by ELISA. BMMOs were stimulated with LPS as a positive control (10 ng/mL). Raw IL-1β concentration was measured from 5 different milk collections (A) and standardised to IL-1β produced from BMMOs stimulated with LPS in each corresponding milk collection (B). Data is presented as the mean ± SD with N = 6. C) For each milk collection, the somatic cell count (SCC) per µL was measured using a DeLaval cell counter on whole milk and cells/µL on obtained isolated cells. The total cell numbers were calculated in 3 L milk and isolated cells in 50 mL PBS. The yield was calculated for the percentage of isolated cells obtained from whole milk. Following the BMMO isolation protocol, the numbers of macrophages/well were calculated using a haemocytometer