Vibrio alginolyticus frequently breaks out in aquatic animal breeding operations involving shrimp, and it can endanger human health through food and wound infections. The antibacterial effect and mechanism of fermented pomegranate peel polyphenols (FPPPs) on V. alginolyticus were investigated. The results indicated that FPPPs had a strong inhibitory effect on the growth of V. alginolyticus, and their minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) were 2 and 4 mg/mL. FPPPs significantly reduced biofilm formation and biofilm metabolic activity in V. alginolyticus, down-regulated the expression levels of lafA, lafK, fliS and flaK genes involved in flagellar synthesis and inhibited swimming and swarming motility (p < 0.05). Meanwhile, under the treatment of FPPPs, the activities of catalase (CAT) and superoxide dismutase (SOD) in V. alginolyticus were significantly reduced, and the levels of reactive oxygen species (ROS) and extracellular malondialdehyde (MDA) were significantly increased (p < 0.05). FPPPs also resulted in a significant increase in alkaline phosphatase (AKP) activity, protein and nucleic acid content, as well as conductivity from V. alginolyticus cultures. Scanning electron microscopy (SEM) images further revealed that V. alginolyticus treated with FPPPs showed leakage of intracellular substances, abnormal cell morphology and damage to cell walls and cell membranes, with the severity of the damage in a clear dose-dependent manner. Therefore, FPPPs can be used as a promising food-grade antibacterial agent, notably in seafood to control V. alginolyticus.
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