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200. The absorbance values of the negative control wells (containing no cells) were subtracted from the values of the test wells. Standard antifungal powders of voriconazole (Pfizer, Groton, CT, [http://en.wikipedia.org/wiki/Thalidomide Thalidomide] USA), amphotericin?B (Sigma, St Louis, MO, USA) and anidulafungin (Pfizer) were obtained from the respective manufacturers. Stock solutions of voriconazole, amphotericin?B and anidulafungin were prepared in dimethylsulphoxide (Sigma). Antifungal agents were then diluted with RPMI-1640 medium (Sigma) and buffered to pH?7.0 with morpholinepropranesulphonic acid buffer (Sigma). The MICs for planktonic cells were determined according to the standard CLSI document M27-A3 [18]. Voriconazole, amphotericin?B and anidulafungin concentrations ranged from 0.03 to 16?mg/L. CLSI breakpoints for susceptibility were used for voriconazole and anidulafungin, and a provisional susceptible breakpoint of ��1?mg/L was used for amphotericin?B. The MICs for sessile [http://www.selleckchem.com/PARP.html check details] cells (biofilms) were determined with the previously described microtitre assay [17]. Mature biofilms were allowed to form in 96-well microtitre plates for 48?h, and the cell densities of the biofilms were estimated using the XTT absorbance assay. The antifungal agents were prepared in a series of two-fold dilutions from 16 to 0.03?mg/L. The inhibitory effects of the antifungals were measured as the optical densities of the antifungal-treated wells relative to those of the control (antifungal-free) wells (considered to be 100%), as determined in the XTT assays, and are expressed as percentages (MIC50 and MIC80) of the values for control wells. All isolates were tested at least twice. The chi-square test was used to compare biofilm positivity between C.?parapsilosis sensu?stricto [http://www.selleckchem.com/products/pci-32765.html selleck screening library] isolates recovered from blood cultures and those recovered from different sources and between different species. Differences between groups were considered to be significant when the p-value was