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Use of natural compounds as antivirulence drugs could be an alternative therapeutic approach to modify the outcome of bacterial infections, particularly in view of growing resistance to available antimicrobials. Here, we show that sub-bactericidal concentration of anethole, a component of sweet fennel seed, could suppress virulence potential in O1 El Tor biotype strains of toxigenic  Vibrio cholerae , the causative agent of the ongoing 7 th  cholera pandemic. The expression of cholera toxin (CT) and toxin coregulated pilus (TCP), the major virulence factors of  V .  cholerae , is controlled through a regulatory cascade involving activation of ToxT with synergistic coupling interaction of ToxR/ToxS with TcpP/TcpH. We present evidence that anethole inhibits  in vitro  expression of CT and TCP in a  toxT -dependent but  toxR/toxS -independent manner and through repression of  tcpP/tcpH , by using bead-ELISA, western blotting and quantitative real-time RT-PCR assays. The cyclic AMP (cAMP)-cAMP receptor protein (CRP) is a well-studied global signaling system in bacterial pathogens, and this complex is known to suppress expression of  tcpP/tcpH  in  V .  cholerae . We find that anethole influences the virulence regulatory cascade by over-expressing  cyaA  and  crp  genes. Moreover, suppression of toxigenic  V .  cholerae -mediated fluid accumulation in ligated ileum of rabbit by anethole demonstrates its potentiality as an antivirulence drug candidate against the diseases caused by toxigenic  V .  cholerae . Taken altogether, these results revealing a mechanism of virulence inhibition in  V .  cholerae  by the natural compound anethole, may have relevance in designing antivirulence compounds, particularly against multiple antibiotic resistant bacterial pathogens.

Suppression of Virulence of Toxigenic Vibrio cholerae by Anethole through the Cyclic AMP (cAMP)-cAMP Receptor Protein Signaling System