Culture and Identification of Candida Albicans from Vaginal Ulcer and Separation of Enolase on SDS-PAGE

Abstract

Candida albicans were isolated from patients with clinical symptoms of vaginal ulcer. Culture test for vaginal swab andscrapings were conducted on Sabouraud’s dextrose broth and Sabouraud’s dextrose agar plate respectively. Hichromecandida agar culture was used for differential identification of Candida. Smears from vaginal scrapings were preparedfor gram staining. The suspected strain of Candida was inoculated on corn meal agar medium for chlamydosporeformation. The suspected strain of Candida was inoculated in human serum for germ tube formation. Carbohydrateassimilation and fermentation tests were also conducted. The selected Candida colony was inoculated in YEPD mediumfor subculture and the cultured organism was harvested. The organisms were homogenized, centrifuged and thesupernatant was filtered. The filtrate was extracted in chloroform. The extract was centrifuged and the aqueous phasewas dialyzed. The dialyzed crude enolase was subjected to SDS-PAGE. The Sabouraud’s dextrose broth inoculatedwith vaginal swab showed turbid growth. The scraping from vagina showed typical smooth creamy white colonies witha characteristic yeast odour on Sabouraud’s dextrose agar plate. On Hichrome candida agar the Candida growthappeared as glistening green colored. Gram stained smears from vaginal scraps showed appearance of fungus as yeast budding. On corn meal agar the suspected Candida growth showed the formation of large, highly refractive, thickwalled terminal chlamydospores. Germ tubes were seen as long tube like projections extending from the yeast cells onhuman serum inoculated with suspected strain of Candida. The carbohydrate assimilation tests were positive fordextrose, maltose, sucrose, galactose, xylose and trehalose, and negative for lactose, melibiose, ellobiose, inositol,reffinose and dulcitol. The carbohydrate fermentation tests showed positive for dextrose, maltose, galactose andtrehalose, and negative for sucrose and lactose. SDS-PAGE for enolase from C. albicans showed a single polypeptideband of around 46 – 48 kDa.