Synbiotics are a combination of probiotic bacteria and the growth promoting prebiotic ingredients that purport "synergism".Probiotics are well known for their ability to restore a healthful balance of commensal microbes supporting the repair of hyper-permeable epithelial barriers and interfering with infection. The present research was designed to evaluate the anti-virulence role of cell-free supernatants of synbiotic cultures.The effect of prebiotics fructo-oligosaccharide, inulin and isomaltose on probiotic Lactobacillus acidophilus, Lactococcus lactis, Lactobacillus casei, Lactobacillus reuteri and Saccharomyces boulardii growth, fermentation products has been investigated. Cell-free supernatants of synbiotics cultures have been studied for their antimicrobial, antibiofilm and anti-adhesion properties. Results highlight that prebiotics can enhance the antimicrobial and anti-virulence activities of probiotics against potential pathogens Staphylococcus aureus and Escherichia coli.The present research supports, for the first time, the value of anti-virulence activity of metabolites produced by synbiotics suggesting their use as a suitable adjuvant in antibacterial treatment.