Effect of ultrasonication and thermal and pressure treatments, individually and combined, on inactivation of Bacillus cereus spores.

Bacillus cereus spores are a concern to the food industry due to their high resistance to processing and their ability to germinate to vegetative cells under suitable conditions. This research aimed to elucidate the mechanisms of Bacillus cereus spore inactivation under ultrasonication (US) combined with thermal (thermosonication, TS) treatments, with pressure (manosonication, MS) treatments, and with thermal and pressure (manothermosonication, MTS) treatments. Electronic microscopy, dipicolinic acid (DPA) release, and flow cytometric assessments were used to investigate the inactivation effect and understand the inactivation mechanisms. The sporicidal effects of the US and thermal treatment were slight, and the MS and TS also showed little inactivation effect. However, ultrasonication promoted the detachment of the exosporium, thereby reducing the spore's ability to adhere to a surface, while the thermal treatment induced a decrease in the electron density in the nucleoid of bacterium, which retained a relatively intact exosporium and coat. MS caused 92.54% DPA release, which might be due to triggering of the germinant receptors or releasing of ions and Ca2+-DPA. In addition, the morphological changes such as core hydration and cortex degradation were significant after treatment with MS. The release of DPA and the morphological changes were responsible for the reduction in thermal resistance. The MTS showed a remarkable inactivation effect of 3.12 log CFU/mL reductions after 30 min of treatment. It was the most effective treatment and exhibited a large fraction of damage. In addition, the MTS had a significant impact on the intracellular structure of the spores, with the coat destroyed and the cortex damaged. These results indicated that ultrasonication combined with thermal and pressure treatments had a significant sporicidal effect on Bacillus cereus spores and could be a promising green sterilization technology.