Sporulation boundaries and spore formation kinetics of Bacillus spp. as a function of temperature, pH and aw

Sporulation niches in the food chain are considered as a source of hazard and are not clearly identified. Determining the sporulation environmental boundaries could contribute to identify potential sporulation niches. Spore formation was determined in a Sporulation Mineral Buffer. The effect of incubation temperature, pH and water activity on time to one spore per mL, maximum sporulation rate and final spore concentration was investigated for a Bacillus weihenstephanensis and a Bacillus licheniformis strain.Sporulation boundaries of B. weihenstephanensis and of B. licheniformis were similar to, or included within, the range of temperatures, pH and water activities supporting growth. For instance, sporulation boundaries of B. weihenstephanensis were evaluated at 5 °C, 35 °C, pH 5.2 and aw 0.960 while growth boundaries were observed at 5 °C, 37 °C, pH 4.9 and aw 0.950. Optimum spore formation was determined at 30 °C pH 7.2 for B. weihenstephanensis and at 45 °C pH 7.2 for B. licheniformis. Lower temperatures and pH delayed the sporulation process. For instance, the time to one spore per mL was tenfold longer when sporulation occurred at 10 °C and 20 °C, for each strain respectively, than at optimum sporulation temperature. The relative effect of temperature and pH on sporulation rates and on growth rates is similar. This work suggests that the influence of environmental factors on the quantitative changes in sporulation boundaries and rates was similar to their influence on changes in growth rate.

► Temperature, pH and aw growth limits are reasonable predictions of sporulation boundaries.
► Temperature and pH determined time to first spore.
► Temperature and pH determined final spore concentration.
► Sporulation rates and growth rates are similarly affected by environmental factors.