From Germination to Stagnation: Sodium Diacetate's Impact on <scp><i>Clostridium perfringens</i></scp> Spores

ABSTRACT Sodium diacetate is recognized for its high efficiency as a preservative, demonstrating strong antibacterial properties that help extend the shelf life of food products. However, it is still unclear how it influences spores. This research investigated the influence of sodium diacetate on the germination and outgrowth stage in the revival process of Clostridium perfringens ( C. perfringens ) spores. The germination and outgrowth rate of spores after sodium diacetate treatment were measured. The changes of spore inner membrane were monitored by laser confocal microscope, electron scanning microscope and macromolecular leakage. The interaction between sodium diacetate and DNA was investigated by EB fluorescence probe, and the finding was verified by the outgrowth of spores in ham sausage. An investigation into the effects of sodium diacetate treatment on the biological activity, inner membrane permeability, morphology, and DNA integrity of spores devoid of both the coat and cortex revealed that sodium diacetate significantly hindered the transition of germinated spores into vegetative cells. In the presence of sodium diacetate, C. perfringens spores underwent germination, characterized by the hydrolysis of the spore cortex and the disassembly of the spore coat. As the spores advanced into the outgrowth phase, sodium diacetate penetrated their structure, causing damage to the inner membrane and compromising DNA integrity. Moreover, sodium diacetate was demonstrated to effectively inhibit spore outgrowth in ham sausage. This study provided theoretical guidance and references for the application of sodium diacetate in food to control the germination and outgrowth of spore‐forming bacteria.