In Vitro Biofilm Activity of Non-Candida albicans Candida Species

Candidosis has been attributed to C. albicans; however, infections caused by non-Candida albicans Candida (NCAC) species are increasingly being recognised. The ability of Candida to grow as a biofilm is an important feature that promotes both infection and persistence in the host. The biofilms' activity is significant since high activity might be associated with enhanced expression of putative virulence factors, whilst in contrast low activity has previously been suggested as a mechanism for resistance of biofilm cells to antimicrobials. The aim of this study was to determine the metabolic activity of in vitro biofilms formed by different clinical isolates of NCAC species. The in situ total metabolic activity of C. parapsilosis, C. tropicalis and C. glabrata biofilms was determined using 2,3-(2-methoxy-4-nitro-5-sulphophenyl)-5-(phenylamino) carbonyl-2H-tetrazolium hydroxide (XTT) reduction assay, and the number of cultivable cells was also established by CFU (colony forming unit) counts. The biofilm structure was assessed by scanning electron microscopy (SEM). Results showed that total biofilm metabolic activity was species and strain dependent. C. glabrata exhibited the lowest biofilm metabolic activity despite having the highest number of biofilm cultivable cells. Similarly, the metabolic activity of resuspended C. glabrata biofilm and planktonic cells was lower than that of the other species. This study demonstrates the existence of intrinsic activity differences amongst NCAC species, which could have important implications in terms of species relative virulence. Furthermore, the absence of an obvious correlation, between cultivable cells number and total biofilm activity, raises the question about which parameter is the most appropriate for the in vitro assessment of biofilms and their potential clinical significance.