| Summary: | Coenzyme Q<sub>0</sub> (CoQ<sub>0</sub>) has anti-inflammatory and anti-tumor effects; however, the antimicrobial and antibiofilm activities of CoQ<sub>0</sub> against <i>Salmonella enterica</i> serovar Typhimurium are unknown. Thus, we investigated the bacteriostatic and antibiofilm activities, along with the underlying mechanism, of CoQ<sub>0</sub> against <i>S</i>. Typhimurium. The minimum inhibitory concentration (MIC) of CoQ<sub>0</sub> against <i>S. enterica</i> serovars Typhimurium was 0.1–0.2 mg/mL (549–1098 µM), and CoQ<sub>0</sub> at MIC and 2MIC decreased viable <i>S</i>. Typhimurium counts below detectable limits within 6 and 4 h, respectively. CoQ<sub>0</sub> at 20MIC (4 mg/mL) reduced <i>S</i>. Typhimurium on raw chicken by 1.5 log CFU/cm<sup>3</sup> within 6 h. CoQ<sub>0</sub> effectively disrupted cell membrane integrity and induced morphological changes in the cell, resulting in hyperpolarization, decreased intracellular ATP concentrations, and cellular constituents leakage. Biofilm-associated <i>S</i>. Typhimurium cells were killed by CoQ<sub>0</sub> treatment. These findings suggest that CoQ<sub>0</sub> could be applied as a natural antibacterial substance for use against <i>S</i>. Typhimurium by the food industry.
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