Caspase of the compounds was measured using agar well diffusion assay against B. subtilis, S. aureus, E. coli and P. aeruginosa. The sterile discs were impregnated with 50 lg per disc test compounds. The antibiotics ceftazidime and ciprofloxacin were used as positive reference standards. Solvent control was also tested along with the test samples. The antibacterial activity was evaluated by measuring the zone of growth inhibition surrounding the discs. All the assays were carried out in triplicate. Minimum inhibitory concentration. MIC was determined by standard macro dilution broth test as recommended by the National Committee for Clinical Laboratory Standards, USA, against all the four test bacteria. The pure compounds and standard antibiotics were tested at final concentrations, prepared from serial twofold dilutions, ranging from 1 to 2000 lg ml1. The MIC value was defined as the lowest concentration of the compound showing no visible growth. Triplicate sets of tubes were maintained for each concentration of the test sample. Minimum bactericidal concentration. MBC was determined MEK signaling pathway according to the method of Smith Palmer et al. 1998 against all the four test bacteria. About 100 ll from the tubes not showing growth were plated on nutrient agar.
MBC is the lowest concentration at which bacteria failed to grow in nutrient broth and nutrient agar inoculated with 100 ll of suspension. Triplicate sets of smoothened pathway tubes were maintained for each concentration of the test sample. Determination of antifungal activity Antifungal activity was determined by the agar disc diffusion method against A. flavus, C. albicans, F. oxysporum, R. solani and P. expansum. The sterile dicks were impregnated with 50 lg per disc of compounds. Amphotericin B was used as reference standard for C. albicans. Bavistin was used as reference standard against other fungi. Diameter of zone of inhibition was measured. All the assays were carried out in triplicate. Minimum inhibitory concentration. MIC was determined using potato dextrose agar media against the standard fungicide bavistin by the poisoned food technique against A. flavus, F. oxysporum, R. solani and P. expansum. A stock solution of 1000 lg ml1 of the test compound was prepared, which was further diluted with methanol to give the required concentrations 1000 1 lg ml1. One tube was used as solvent control. For C. albicans, the broth dilution method was adopted celecoxib using potato dextrose broth against the standard fungicide amphotericin B.
All experiments were in triplicate for each treatment against each fungus. Results Isolation and characterization of bioactive compounds The ethyl acetate fraction of the cell free culture filtrate of the bacteria showed antibacterial activity againstAntibacterial activity. The isolated compounds were tested for antibacterial activity against four bacterial bruker strains using standard methods. MIC and MBC values were also determined. From the results of the well diffusion assay, compound 1 presented antibacterial activity against all tested Gram positive and negative bacteria. MIC for all the test bacteria are shown in Table 2. The micro organism that presented highest sensitivity towards resveratrol was B. subtilis 1, followed by S. aureus and E. coli, which have presented a MIC of 32 lg ml1.