Ultrasound combined with FeSO4 facilitated the occurrence of ferroptosis in Vibrio parahaemolyticus

Ultrasound (US) as a sustainable non-thermal sterilization technology that is employed either independently alone or in combination with other processing methods to eliminate food-borne pathogens in the food industry. In the present study, the synergistic effects of US combined with FeSO₄ against Vibrio parahaemolyticus were investigated. The results demonstrated that the combination of ultrasound and FeSO₄ had an excellent bactericidal activity on V. parahaemolyticus. Treatment with US (100 W) and FeSO₄ (8 μM) for 15 min could kill more than 99.9 % cells. Furthermore, the observed cell death was identified as classical ferroptosis, characterized by ferroptosis hallmarks including iron-dependent, ROS burst, membrane damage and lipid peroxide accumulation. Addition of ferroptosis inhibitor liproxstatin-1 alleviated the cell death induced by the combination treatment. Transcriptome analysis further revealed that the US-FeSO₄ treatment significantly influenced pathways related to fatty acid metabolism, ferroptosis, biofilm formation, RNA degradation, oxidative phosphorylation and other key processes, which likely contributed to the occurrence of ferroptosis. Based on these findings, we speculated that cavitation effect of US promoted the entry of Fe²⁺, leading to the generation of free radicals primarily responsible for ferroptosis by US-FeSO₄. Taken together, this study provides valuable insights into the biological pathway involved in ultrasound sterilization and presents an alternative strategy to eradicate microorganism in food products.