Figure 2. The effect of spore production environment on subsequent germination of C. sporogenes spores.

For germination tests, spores were incubated in 20 mM Tris buffer, pH 7.4, L-alanine (100 mM), L-lactate (50 mM) and NaHCO₃ (50 mM) at 30°C. Spores were produced in CMB at different temperatures; 15°C, 20°C, 28°C, 30°C, 37°C. Spores were also produced in TY broth at 37°C and on RCM+SM plates at 37°C. Spores produced at 37°C in CMB and incubated in 20 mM Tris, pH 7.4, L-lactate (50 mM) and NaHCO₃ (50 mM) only, were included as a negative control (WT Buffer). Spore germination was confirmed by phase contrast microscopy. Data labels (right) refer to percentage germination observed by phase contrast microscopy at the end of the experiment. Error bars represent the standard deviation of 3 independent experiments.