Assay for haemolytic activity
Haemolytic and haemoxidative activity of M. bovis was determined as described previously (Großhennig et al., 2016). Briefly, harvestedM. bovis cells were resuspended in PBS and added to washed sheep RBCs (final concentration: 2% in PBS) with, or without, supplements in a final volume of 1 ml. Aliquots without cells served as control. The test samples were incubated in a rotary shaker at 110 r.p.m. and 37oC for time periods. After 10 min centrifugation at 480C and 1400 x g , the supernatant was transferred into a new tube and the pellet was resuspended in 1 ml dH2O. The supernatant and pellet fractions were photographed, and their spectra were recorded from 370 to 700 nm.