Inactivation of dried spores of Bacillus subtilis 168 by a treatment combining high temperature and pressure.

Specific treatments combining high temperatures of up to 150 °C and moderate pressure of up to 0.6 MPa have been applied to Bacillus subtilis 168 spores conditioned at different aw levels (between 0.10 and 0.70) corresponding to different residual water contents within the spore core. The spores were treated as a dry powder in a pressurized nitrogen environment or in water/glycerol solutions. These thermodynamic conditions were intended to prevent any water evaporation from the spore core during time/temperature treatments. Our results clearly show that retaining liquid water in the core by applying pressure during the treatment resulted in greater spore destruction (between 2.4 and 4.9 log at 150 °C, 120 s and aw 0.5 in powder) than the destruction observed after the treatment at atmospheric pressure (0.7 log), during which the water rapidly evaporated because its boiling point was reached. Moreover, we found that the water activity level of the spore had a significant impact on spore destruction: the higher the aw level, the greater the spore inactivation. We obtained similar results from spores heat-treated in powder and in water/glycerol solution at the same aw, confirming the strong influence of this parameter. We hypothesized that the increased spore inactivation was related to the well-known thermal sensitivity of vital organic molecules such as proteins, enzymes, and ribosomes in the presence of water.

Extremely rapid acclimation of Escherichia coli to high temperature over a few generations of a fed-batch culture during slow warming.

This study aimed to demonstrate that adequate slow heating rate allows two strains of Escherichia coli rapid acclimation to higher temperature than upper growth and survival limits known to be strain-dependent. A laboratory (K12-TG1) and an environmental (DPD3084) strain of E. coli were subjected to rapid (few seconds) or slow warming (1°C 12 h(-1)) in order to (re)evaluate upper survival and growth limits. The slow warming was applied from the ancestral temperature 37°C to total cell death 46-54°C: about 30 generations were propagated. Upper survival and growth limits for rapid warming (46°C) were lower than for slow warming (46-54°C). The thermal limit of survival for slow warming was higher for DPD3084 (50-54°C). Further experiments conducted on DPD3084, showed that mechanisms involved in this type of thermotolerance were abolished by a following cooling step to 37°C, which allowed to imply reversible mechanisms as acclimation ones. Acquisition of acclimation mechanisms was related to physical properties of the plasma membrane but was not inhibited by unavoidable appearance of aggregated proteins. In conclusion, E.coli could be rapidly acclimated within few generations over thermal limits described in the literature. Such a study led us to propose that rapid acclimation may give supplementary time to the species to acquire a stable adaptation through a random mutation.