were used to determine the MIC of nisin and reutericyclin against
germination of endospores or growth vegetative cells ofClostridium
spp. MIC procedures were carried out in an anaerobic hood. In brief,
100 ml of RCM broth was added to each well of a microtitre plate.
Nisin or reutericyclin stock solutions (100 ml) were added to separate wells and serially diluted. Stationary phase cultures of Clostridium spp. or endospore suspensions with an OD600 2.0 were
diluted to one-tenth of the initial concentration in RCM broth, and
microtitre plates were inoculated with 50 ml of the diluted culture.
Plates were incubated anaerobically at 37
C for 24 h (vegetative
cells) or 6 d (endospores).