Thymoquinone (TQ) has been demonstrated to have anti-cancer, anti-inflammatory, antioxidant, and anti-diabetic activities. Shigella flexneri is the main pathogen causing shigellosis in developing countries. In this study, the antibacterial activity of TQ against S. flexneri and its possible antibacterial mechanism were studied. In addition, the inhibitory effect of TQ on the formation of S. flexneri biofilm was also investigated. The results showed that both the minimum inhibitory concentration and the minimum bactericidal concentration of TQ against S. flexneri ATCC 12022 were 0.2 mg/mL. After treatment with TQ at 0.4 mg/mL in Luria-Bertani broth for 3 h, or treatment with 0.2 mg/mL TQ in phosphate-buffered saline for 60 min, the number of S. flexneri (initial number is 6.5 log colony-forming units/mL) dropped below the detection limit. TQ also displayed good antibacterial activity in contaminated lettuce juice. TQ caused an increase in intracellular reactive oxygen species level, a decrease in intracellular adenosine triphosphate (ATP) concentration, a change in the intracellular protein, damage to cell membrane integrity and changes in cell morphology. In addition, TQ showed the ability to inhibit the formation of S. flexneri biofilm; treatment resulted in a decrease in the amount of biofilm and extracellular polysaccharides, and the destruction of biofilm structure. These findings indicated that TQ had strong antimicrobial and antibiofilm activities and a potential to be applied in the fruit and vegetable processing industry or other food industries to control S. flexneri.
Keywords: Shigella flexneri; antibacterial mechanism; biofilm; lettuce juice; thymoquinone.