gal psi SS pall alk) qa cise 9 513! apg Sealy Azesplall Gla hall ¢5 goal) ppl! :3405 1592: (4) cies NRC/VPRAISUDI/F 06) 8 adits dene/a CS Julad yt Materials & Methods Materials: Nutrient agar medium & sterile Petri dishes Table No. (1), physical properties of samples Teva aI: Gaal a3 TWewnyre/a) aay eae LY: Gnas laa) NRC/VPRA/SUD/F 06 Saas 1) vyNutrient agar medium(fluka, code:70148):- A general culture medium for less fastidious microorganisms as well as for permanent cultures. Composition: kre gsi Ingredients : z Yeast extract 2.0 Peptone 5.0 Meat extract 1.0 NaCl 5.0 Agar 15.0 pH 7.4 + 0.2 prepared media: Store below 8°C, protected from direct light. Store dehydrated powder, in a dry place, in tightly-sealed containers at 2-25°C. Directions: Suspend 28 g in 1 liter of distilled water. Bring to the boil to dissolve completely. Sterilize by autoclaving at 121°C for 15 minutes. Preparation of Inoculum The inoculum should be adjusted so that 108 cfu/ml are applied to the plates. for preparing the desired inoculum byMcFarland, J., (1907): The nephelometer: an instrument for estimating the number of bacteria in suspensions used for calculating the opsonic index and for vaccines. J. Am. Med. Assoc. 49: 1176-1178. Mostafa, A. F., Abd El Aty, A., A., Hamed, K., E., Eid, M., B. and Ibrahim, A., N. (2016): Enzymatic, kinetic and anti-microbial studies on Aspergillus terreus culture filtrate and Allium cepa seeds extract and their potent applications, Biocatalysis and Agricultural Biotechnology, 5:116-122. S. Lapage, J. Shelton, T. Mitchell. (1970): Methods in Microbiology, J. Norris, D. Rippons (Eds.), 3A, Academic Press, London. Zamora, L.L & Pe rez-Gracia, M.T., (2012): Using digital photography to implement the McFarland method. R. Soc. Interface.9,1892-1897.El-serwy, WS., et al., (2015. ): Synthesis of New Benzofuran Derivatives and Evaluation of their Antimicrobial Activities. Research Journal of Pharmaceutical, Biological and Chemical Sciences. 6: 213-224. Frey, F.M & Meyers., (2010):Antibacterial activity of traditional medicinal plants used by Hauden osaunee peoples of New York State, BMC Complementary and Alternative Medicine.:10-64 Graziano, S., T., Cuzzullin C., M., Franco, C., G., Schwartz-Filho, O., H., de Andrade, D., E., Groppo, C., F. and Cogo-Miiller, K. (2015): Statin Antimicrobial Effects: Simvastatin as a Potential Drug against Staphylococcus aureus Biofilm. PLOS ONE, 10:7-10. J. MacFaddin, (1985): Media for Isolation-Cultivation-Identification- Maintainance of Medical Bacteria, Vol. 1, Williams, Wilkins, Baltimore. Kim, K., Sung, S., W., Suh, K., B., Moon, S., Choi, J., Kim, G., J. and Lee, G., D., (2009): Antifungal activity and mode of action of silver nano-particles on Candida albicans. Biometals. 22:235-242. M.A. Sagardoy, C.M. Salerno. (1984): Studies on heterotrophic bacteria in some Argentine soils, Anal.Edaf.Agrobiol.42: 2069. MLL. Gray, HJ. Stafseth, F. Thorp. (1950): The use of potassium tellurite, sodium azide and acetic acid in a selective medium for the isolation of listeria monocytogenes, J. Bact., 59: 443. McDermott, F., P., Walker D., R. and White, G., D. (2002): Antimicrobials: Modes action and Mechanisms of Resistance, International Journal of Toxicology, 22:135-143.Result: ‘Table (2): Inhibition zone diameter (millimeter) of the samples Samples Blank Treated sample sample Test bacteria 1 2 3 1- | Pseudomonas aeruginosa NIL 26.0 25.0 24.3 | Ds | Staphyllococus aureus NIL 25.0 29.0 26.0 3- Candida albicans NIL 21.0 22.0 22.0 | * Nil: No antimicrobial activity recorded. 800808 REFERENCES Barry, A.L., (1980): Procedure for testing antimicrobial agent in agar media. In V Lorianed) Antibiotica in laboratory medicines. Willims and Wilkins Co. Baltimore.:1-23 Eaton A. D., Clesceri L. S. and Greenberg A. E., (Ed.), (1998): Standard Methods for the Examination of Water and Waste water, 20th Ed., American Public Health Association. Washington, D.C. El-sersy & Abou-Elela, (2006).: Antagonistic effect of marine Nocardia brasiliensis against the fish pathogen Vibrio damsela: Application of Plackett-Burman experimental design to evaluate factors affecting the PRE3- Pathogenic Yeast: Candida albicans (ATCC 10231). Methods: 1- Qualitative evaluations were carried out in nutrient agar plates according to (Mostafa et al., 2016). 2- Bacteria used in this study were Gram positive bacteria [Staphylococcus aureus (ATCC 6538)], Gram negative bacteria [Pseudomonas aeruginosa (ATCC27853)] and pathogenic yeast [(Candida albicans ATCC 10231)]. 4-The inoculation of all microorganisms was prepared from fresh overnight broth cultures that were incubated at 37°C (El-serwy et al., 2015). 5-The inoculum size of these pathogenic strains was prepared and adjusted to approximately 0.5 McFarland standard (1.5 x 108 /ml), 25.0 ul of both Bacterial & yeastal suspensions were inoculated into each plate containing 20.0 mL of the sterile nutrient agar medium (NA). 6- After the media cooled and solidified, the prepared samples were applied on the surface of that inoculated agar plates which prepared previously 6- These seeded plates were placed in the refrigerator for one hour, followed by incubation at 37 °C for 24 hrs and zones of inhibition (ZI) were measured in mm. (Mostafa et al., 2016) and tabulated in the following table no. (2).