In the present study bioefficacy and biocidal activity of Terminalia chebula fruit water extract against common infectious bacteria; Staphylococcus aureus, Bacillus subtilis and mixed culture of S.aureus and B.subtilis (1:2) was assessed. The characterization and detection of biofilm formation by these bacteria was analyzed using common methods: Tissue culture plate method (TCP), Congo red agar method (CRA). The extract in three different concentrations 1%, 2% and 5% were used for optimization of biocide concentration using TCP method. The results from both detection methods indicated the effective biocidal activity of the extract on biofilm formation. The optimum extract concentration for reducing or inhibiting the biofilm formation in both the species was found to be 2%. The results suggests the use of the fruit extract as an biological alternative for eradication of biofilm formation in medical devices in place of commonly used hospital chemical biocides.