Inactivation Effect of Germination Combined with Cold Plasma Treatment on Bacillus licheniformis Spores

Food spoilage, primarily caused by spore-forming bacteria, has become a critical concern since it results in substantial economic losses within the food industry. Past investigations have successfully identified Bacillus licheniformis as the main bacterium responsible for spoilage in roast chicken. In this study, we screened a new sterilization combination from 16 germinants and 4 cold plasma conditions, respectively. Among them, the combination of "A"GFNa-1 (composed of 60 mmol/L L-alanine, 10 mmol/L D-glucose, 10 mmol/L D-fructose, and 1 g/L NaCl) with cold plasma treatment (packed with 100% argon at 70 kV) proved effective in deactivating B. licheniformis spores, resulting in a reduction of approximately 2.1 log CFU/mL. Furthermore, we exposed the spores to different conditions: CK (no germination, no cold plasma), MF (germination only), CP (no germination, 100% argon packed, 70 kV cold plasma treatment for 3 min), and MF + CP (germination for 5 h, 100% argon packed, 70 kV cold plasma treatment for 3 min). The results of heat inactivation and dipicolinic acid (DPA) release rate demonstrated that cold plasma treatment effectively inactivated both spores and vegetative cells without inducing germination. Additionally, the reduced survival under hyperosmotic conditions and the presence of distinct red fluorescence patterns observed through confocal laser scanning microscopy (CLSM) collectively suggest that cold plasma treatment disrupts the inner membrane structure and leads to the inactivation of B. licheniformis. Overall, our findings indicate a spore clearance rate of 99.2% and suggest that the combination of efficient germinants and cold plasma treatment holds promise as a viable approach to mitigate spore contamination in the food industry.