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

Candida albicans were isolated from patients with clinical symptoms of vaginal ulcer. Culture test for vaginal swab and scrapings were conducted on Sabouraud’s dextrose broth and Sabouraud’s dextrose agar plate respectively. Hichrome candida agar culture was used for differential identification of Candida. Smears from vaginal scrapings were prepared for gram staining. The suspected strain of Candida was inoculated on corn meal agar medium for chlamydospore formation. The suspected strain of Candida was inoculated in human serum for germ tube formation. Carbohydrate assimilation and fermentation tests were also conducted. The selected Candida colony was inoculated in YEPD medium for subculture and the cultured organism was harvested. The organisms were homogenized, centrifuged and the supernatant was filtered. The filtrate was extracted in chloroform. The extract was centrifuged and the aqueous phase was dialyzed. The dialyzed crude enolase was subjected to SDS-PAGE. The Sabouraud’s dextrose broth inoculated with vaginal swab showed turbid growth. The scraping from vagina showed typical smooth creamy white colonies with a characteristic yeast odour on Sabouraud’s dextrose agar plate. On Hichrome candida agar the Candida growth appeared as glistening green colored. Gram stained smears from vaginal scraps showed appearance of fungus as yeast International Journal of Biology January, 2010 85 budding. On corn meal agar the suspected Candida growth showed the formation of large, highly refractive, thick walled terminal chlamydospores. Germ tubes were seen as long tube like projections extending from the yeast cells on human serum inoculated with suspected strain of Candida. The carbohydrate assimilation tests were positive for dextrose, 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 and trehalose, and negative for sucrose and lactose. SDS-PAGE for enolase from C. albicans showed a single polypeptide band of around 46 – 48 kDa.