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IJHSR. 2014; 4(11): 68-76


A Study of Biofilm Production and Antifungal Susceptibility of Clinical Isolates of Candida Species.

Rahul P. Dhale, M. V. Ghorpade, C. A. Dharmadhikari.

Abstract
Purpose: The biofilm formation of an organism can be considered as virulence factor, which turns sensitive organisms into the resistant one for antimicrobial agents. Candida biofilms are observed in blood, mucosal surface and most medical devices, such as stents, shunts, implants, endotracheal tubes, pacemakers, and various types of catheters i.e. nonliving objects in patientís body. This study was designed to characterise speciation of Candida, biofilm production and antifungal activity after biofilm formation.
Materials and Methods: Speciation of Candida was done by Dalmau plate technique on corn meal agar, also sugar assimilation and fermentation test were performed by using 2% concentration of sugars. Quantitative measurement of biofilm formation was assessed by microtitre plate assay for 425 Candida isolates using XTT {2, 3-bis (2-methoxy-4-nitro-5-sulfophenyl)-5-[(phenylamino) carbonyl]-2H-tetrazolium hydroxide} reduction assay. Antifungal susceptibility was done to biofilm forming and non biofilm forming Candida using Percentage Transmission (%T) assay and confirmation by XTT reduction assay.
Results: Out of 425 strains, XTT reduction assay gave 72 biofilm positive strains accounting for 16.94 %. Out of 72, 56 strains became resistant to amphotericin B and 41 strains became resistant to fluconazole after induced biofilm production which is significant. Conclusion: The biofilm formation of an organism can be considered as virulence factor, which turns sensitive organisms into resistant organisms for antimicrobial agents. Candida biofilm formation is observed in blood, mucosal surface and most medical devices in patientís body.

Key words: Biofilm, XTT reduction assay, antifungal resistance, medical devices.



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