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Enhanced Recovery of Stachybotrys chartarum from Environmental SamplesR. A. Billups, K. S. Tilton, Ph.D., and P. S. Warden, Analytical Services, Inc. Recovery of Stachybotrys chartarum (atra) from environmental samples can be difficult due to competition from other more aggressive fungi even on selective media such as Cellulose Agar. S. chartarum ATCC 9182, ATCC 34915, along with Stachybotrys spp. isolated from environmental samples and common competing fungi (Aspergillus flavus, A. niger, Cladosporium brevitomosum, and Penicillium chrysogenum) were grown as mixed cultures in the laboratory under experimental conditions to optimize the procedures used for Stachybotrys spp. recovery. Potato Dextrose Agar (PDA) was chosen as the experimental media based on previous studies at Analytical Services, Inc. which demonstrated that PDA allowed more rapid expression of diagnostic Stachybotrys morphology than Cellulose Agar, Corn Meal Agar, and Malt Extract Agar. Trials were completed using Amphotericin B, Cycloheximide, Miconazole, and Chloramphenicol added at various concentrations to PDA to suppress fungal and bacterial competition. Cultures were incubated at 22°C and 35°C for up to twelve (12) days. Miconazole was most effective at inhibiting growth of the selected non-target fungi while allowing Stachybotrys growth. Characteristic, recognizable S. chartarum colonies were observed at both temperatures, but development was notably faster at 22°C. None of the competitive fungi grew at either temperature on the specialized agar; however, only a limited number of fungal types were tested. The combination antibiotic/antimycotic modified PDA and incubation at 35°C appears to effectively reduce competition and allow enhanced recovery of Stachybotrys spp. Presented at the American Industrial Hygiene Conference and Exposition, June 5-11, 1999, Toronto, ON, Canada |