Vibrio cholerae, a causative agent of the waterborne disease cholera, still threatens a large proportion of world's population. The role of biofilm formation in V. cholerae pathogenesis is well established, as it provides the bacterium enhanced tolerance to antimicrobial agents and increased transmission. In the present study, four medicinal plants used in traditional medicines with antidiarrheal properties were evaluated for its antibiofilm activity. Methanol extracts of these plants (Centella asiatica, Elephantopus scaber, Camellia sinensis, and Holarrhena antidysenterica) showed promising antibiofilm activity against V. cholerae with crystal violet and air-liquid interface coverslip assays. Results revealed that C. asiatica, E. scaber, C. sinensis, and H. antidysenterica extracts significantly inhibited biofilm formation by approximately 75, 76, 78, and 55% at concentrations of 3, 2, 1, and 0.6 mg/mL, respectively. A promising antibiofilm activity of ∼89% inhibition at 1.5 mg/mL concentration was observed when a combination of E. scaber and C. sinensis was used. The herbal extracts were thermostable at a temperature range of 40 to 100°C. The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay revealed that the viability of bacteria was not affected by treatment with these plant extracts. Gene expression studies revealed that extracts of H. antidysenterica leaf, H. antidysenterica bark, and the whole plant of E. scaber and C. asiatica down-regulate aphA or aphB, the major regulator genes modulating both virulence and biofilm formation. Hence, we propose that these herbal combinations could serve as a multifaceted approach to combat the pathogen and also, in turn, reduce antimicrobial resistance development.

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