SABORAUD DEXTROSE AGAR + CHLORAMPHENICOL + GENTAMICIN CAT Nº: 2059 For the selective cultivation and isolation of yeasts and molds FORMULA IN g/l Saboraud Dextrose agar 65,0 Chloramphenicol 0,5 Gentamicin 0,05 Final pH 5.6 ± 0.2 at 25ºC PREPARATION Suspend 65,55 grams of medium in 1 liter of distilled water. Mix well and dissolve by heating with frequent agitation. Boil for one minute until complete dissolution. Sterilize in autoclave at 118ºC for 15 minutes. Cool to 50º, mix well and dispense into plates. The prepared medium should be stored at 8-15°C. The color is amber, slightly opalescent. The dehydrated medium should be homogeneous, free-flowing and beige in color. If there are any physical changes, discard the medium. USES SABOURAUD DEXTROSE AGAR + CHLORAMPHENICOL + GENTAMICIN is a selective medium that can be used for the cultivation of yeast, molds and aciduric microorganisms. It is used for cultivating pathogenic fungi, particularly those associated with skin infections (dermatophytes). Fungal infections have become increasingly more common, especially in the setting of immunocompromised patient. It is imperative first that the etiologic agent be grown and isolated on culture; only after that can the organism be identified and the most proper treatment be ensured. Sabouraud dextrose agar (Cat. 1024), the original formulation or the Emmons modification, without any antibiotic has historically been used as the standard medium for the primary isolation of fungus and is still widely used. This medium is a modification enriched with Gentamicin and Chloramphenicol which supports the growth of a wide range of fungi and, due to its antibiotic content, inhibits the great majority of bacterial contaminants. Dextrose contained in Saboraud dextrose is the fermentable carbohydrate providing carbon and energy. Peptone mixture provides nitrogen, vitamins, minerals and amino acids essential for growth. Bacteriological agar is the solidifying agent. The high dextrose concentration and acidic pH make this medium selective for fungi. Chloramphenicol is an antibiotic which aids in isolating pathogenic fungi from heavily contaminated material, as it inhibits most contaminating bacteria. It is a recommended antibiotic for use with media due to its heat stability and wide bacterial spectrum. Gentamicin is an aminoglycoside antibiotic, used to treat many types of bacterial infections by inhibiting its growth. MICROBIOLOGICAL TEST The following results were obtained in the performance of the medium from type cultures after incubation at a temperature of 20-25 ± 2ºC less than 5 days. Microorganisms Growth Candida albicans ATCC 10231 Good Saccharomyces cerevisiae ATCC 9763 Good Aspergillus brasiliensis ATCC16404 Good Penicillium chrysogenum ATCC 9478 Good Trichophyton mentagrophytes ATCC 24957 Good *Staphylococcus aureus ATCC 25923 Inhibited *Escherichia coli ATCC 25922 Inhibited *Incubate at 35 ± 2ºC for 48 hours BIBLIOGRAPHY Murray, P.R., E.J. Baron, J.H. Jorgensen, M.A. Pfaller, and R. H. Yolken (ed.). 2003. Manual of clinical microbiology, 8th ed. American Society for Microbiology, Washington, D.C Scognamiglio, T., Pramod , R. Davise G., Larone H. Comparison of Inhibitory Mold Agar to Sabouraud Dextrose Agar as a Primary Medium for Isolation of Fungi J Clin Microbiol. 2010 May; 48(5): 1924–1925 Brun, S., J. P. Bouchara, A. Bocquel, A. M. Basile, N. Contet-Audonneau, and D. Chabasse. 2001. Evaluation of five commercial Sabouraud gentamicin-chloramphenicol agar media.Eur. J. Clin. Microbiol. Infect. Dis. 20:718-723. Gray, L. D., and G. D. Roberts. 1988. Laboratory diagnosis of systemic fungal diseases. Infect. Dis. Clin. North Am. 2:779-803 Merz WG, Sandford G, Evans GL. Clinical evaluation of the addition of gentamicin to commercially prepared mycological media J Clin Microbiol. 1976 May;3(5):496-500. STORAGE 25ºC Once opened keep powdered medium closed to avoid hydration. 2ºC