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Effect of medium composition on results of macrobroth dilution antifungal susceptibility testing of yeasts

A total of 62 different clinical yeast isolates were examined for susceptibility to four antifungal agents, amphotericin B, 5-fluorocytosine (5-FC), ketoconazole, and miconazole, using synthetic amino acid medium-fungi (SAAM-F), buffered yeast nitrogen broth (BYNB), Kimmig broth, casein-yeast-glucose broth (CYG), antibiotic medium 3-FDA (ANTI-3), and tryptic soy broth (TSB). A macrobroth dilution format was used with MICs determined after incubation for 24 and 48 h. All analyses were performed in duplicate. In general, MICs were more reproducible after 48 h of incubation. Furthermore, with certain medium-antifungal agent combinations, MICs determined after incubation for 48 h were significantly higher than those determined after 24 h. For instance, with 5-FC irrespective of the medium used, greater than 25% of all 48-h MICs were more than one twofold dilution higher than the corresponding MICs determined after incubation for 24 h. Similar observations were made with amphotericin B when tested with BYNB and CYG and with the imidazoles when tested in all of the media except CYG. The actual MICs obtained with the different antifungal agents were clearly influenced by the test medium used. The rank order of amphotericin B MICs according to test medium was as follows: BYNB greater than SAAM-F = Kimmig = CYG = ANTI-3 = TSB. With 5-FC, the following pattern was observed: Kimmig = ANTI-3 greater than SAAM-F = CYG = TSB greater than BYNB. For both imidazoles, ketoconazole and miconazole, the rank order of MICs according to test medium was BYNB = Kimmig = CYG = ANTI-3 = TSB greater than SAAM-F. The results of this investigation suggested that broth dilution susceptibility testing of yeasts is best performed with an incubation period of 48 h. Furthermore, medium composition can significantly influence the results of such testing.