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• Ferrous sulfate induces ferroptosis in Escherichia coli O157:H7. • Droplet size and PDI of CALNO were effectively decreased after US treatment. • FeSO 4 caused ROS eruption and lipid peroxidation in E. coli O157:H7. • FeSO 4 + CALNO had a synergistic effect on the inactivation of E. coli O157:H7. • FeSO 4 + CALNO significantly reduced E. coli on hami melon and cherry tomato. The purpose of this study was to investigate ferroptosis in Escherichia coli O157:H7 caused by ferrous sulfate (FeSO 4) and to examine the synergistic effectiveness of FeSO 4 combined with ultrasound-emulsified cinnamaldehyde nanoemulsion (CALNO) on inactivation of E. coli O157:H7 in vitro and in vivo. The results showed that FeSO 4 could cause ferroptosis in E. coli O157:H7 via generating reactive oxygen species (ROS) and exacerbating lipid peroxidation. In addition, the results indicated that FeSO 4 combined with CALNO had synergistic bactericidal effect against E. coli O157:H7 and the combined treatment could lead considerable nucleic acids and protein to release by damaging the cell membrane of E. coli O157:H7. Besides, FeSO 4 combined with CALNO had a strong antibiofilm ability to inhibit E. coli O157:H7 biofilm formation by reducing the expression of genes related on biofilm formation. Finally, FeSO 4 combined with CALNO exhibited the significant antibacterial activity against E. coli O157:H7 in hami melon and cherry tomato. [ABSTRACT FROM AUTHOR]