The leaves of C. viminalis were collected from the decorated trees in the city of Fasa in April 2015. The plant identity was confirmed in Department of Botany, Darab Payame Noor University, Iran. After cleaning, leaves …show more content…
The base of this method is transfer of anti-bacterial material into the disc. In this method, at first, sterile blank disc (manufactured by Patton medicine) is put in a solution of extract and after soaking, the discs were picked out of the solution and dried in an oven. The bacterial suspension were prepared of bacterial strains and they were equivalent to 0.5 McFarland (108×1.5 CFU/ml). Then, a uniform cultivation was done by sterile cotton swabs soaked in suspension on the surface of Mueller-Hinton agar (Merck, Germany). Then, the disc containing the extract with concentrations of 500 μg/ml, 250 μg/ml and 125 μg/ml, was removed with sterile forceps and placed in a certain distance from each other, from the edge of the plates, on the surface of Mueller. The plates in an upside down state were incubated at 37 °C for 24 hours, and the diameter of inhibition zone around the disc were measured and sensitivity and resistance of strains were determined and then results were compared with CLSI standard table.20 In each experiment, a disc containing the solvent dimethyl sulfoxide (DMSO) 10% was determined as a negative control and a disc containing Vancomaycin 30 μg/disc and Ciprofloxacin 5 μg/disc standard positive control was used as an antibiotic. The tests were repeated three times and the results were presented in the …show more content…
The examination of synergistic and decreasing effects of extracts on antibiotic is done under inhibitory concentrations (sub-MIC). Concentration of the extracts under inhibitory state to the Mueller-Hinton agar is added with a ratio of 1 to 2 (1 unit extract and 2 unit Mueller-Hinton Agar medium) were used as plate test. Bacterial suspension, with a concentration of 0.5 McFarland (108×1.5 CFU/ml), is cultured on Mueller-Hinton Agar medium containing inhibitory concentrations of the extract in a grass-like state. Antibiotic discs was placed on the surface of the medium. The disc containing solvent is determined as negative control and a disc with extract and an inhibitory concentration (sub-Mic) was recorded as positive control. Then diameter of zone growth inhibition was measured after 24 hours of incubation at 37 °C. To study on the interactions effect between of methanol extract of bottlebrush and antibiotic, was used Vancomaycin 30 μg/disc and Ciprofloxacin 5 μg/disc antibiotic disc.22
Statistical Analysis: experiments were based on a repeated triple testing. The results were averaged and standard deviation of the changes was estimated. A comparison of the antibacterial activity of the samples with standard antibiotics was evaluated by applying t-test and ANOVA