Nowadays, antimicrobial resistance and foodborne burden are a prevalent global concern for human health and sustainable development. In this context, this study aimed to investigate the effect of Bacopa monnieri (L.) Pennell extract against pathogenic foodborne bacteria (Escherichia coli, Staphylococcus aureus, and Listeria monocytogenes). Antibiogram phenotypic analysis on these strains revealed a high multiple antibiotic resistance (MAR) index, which varies from 0.5 to 0.75, and antibiotic resistance of 50-80 per cent. Polarity-wise extracted ethanolic extract (BM04-ET) showed potent anti-listerial (22.33 ± 1.15 mm) and anti-escherichial (20.33 ± 0.58 mm) potential with strong bactericidal activity against tested strains. In vitro phytochemical experiments confirmed a very high content of phenols (214.2 ± 1.79 mg/g and flavonoids (61.01 ± 0.48 mg/g) in BM04-ET
further confirmation via UPLC elucidated the abundance of natural bioactive flavanonol, i.e., dihydroquercetin (224.932 μg/mL), possibly imparting bactericidal property to extract. Optimization studies identifies the best parametric combination with pH value of 6.8, temperature 37 °C, and incubation period of 24 h for the antibacterial effect of BM04-ET. Due to high polyphenolic content and superior antibacterial excellence, the study suggested B. monnieri ethanolic extract could be used as a choice in combating foodborne infection, and further research exploration on key antibacterial components turn out to be a remarkable milestone in food and medicinal industries.