Effects of Na⁺ adaptation on Bacillus cereus endospores inactivation and transcriptome changes

As Bacillus cereus endospores exist in various vegetables grown in soil, the possibility of contamination in food products with high salt concentrations cannot be ignored. Recent studies revealed that harsh conditions affect the resistance of bacteria; thus, we investigated the developmental aspect of heat resistance of B. cereus after sporulation with high NaCl concentration. RNA sequencing was conducted for transcriptomic changes when B. cereus endospores formed at high salinity, and membrane fluidity and hydrophobicity were measured to verify the transcriptomic analysis. Our data showed that increasing NaCl concentration in sporulation media led to a decrease in heat resistance. Also, endospore hydrophobicity, membrane fluidity, and endospore density decreased with sporulation at higher NaCl concentrations. When the transcript changes of B. cereus sporulated at NaCl concentrations of 0.5 and 7% were analyzed by transcriptome analysis, it was confirmed that the NaCl 7% endospores had significantly lower expression levels (FDR<0.05) of genes related to sporulation stages 3 and 4, which led to a decrease in expression of spore-related genes such as coat proteins and small acid-soluble proteins. Our findings indicated that high NaCl concentrations inhibited sporulation stages 3 and 4, thereby preventing proper cell maturation in the forespores and adequate formation of the coat protein and cortex. This inhibition led to decreased endospore density and hydrophobicity, ultimately resulting in reduced heat resistance.resistanceWe expect that this study will be utilized as a baseline for further studies and enhance sterilization strategies.