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.