title: Optimal growth conditions for the determination of the antifungal susceptibility of three species of dermatophytes with the use of a microdilution method.
abstract: As a prerequisite to standardization of dermatophyte susceptibility testing, conditions that support optimal growth of different dermatophyte species must be established. Eighteen isolates of Trichophyton spp. (T rubrum, T mentagrophytes, T tonsurans) were grown in 4 different media: RPMI 1640 with L-glutamine, without sodium bicarbonate and buffered at pH = 7.0; antibiotic medium #3 (Penassay); yeast nitrogen base with 0.5% dextrose buffered at pH = 7.0; and Sabouraud dextrose broth. Incubation for 6 days at 35 degrees C produced the following results: RPMI and Sabouraud dextrose supported equally sufficient growth for all strains tested; Penassay supported growth of only 33% of the isolates tested, and buffered yeast nitrogen base did not support growth of any isolates. RPMI was selected as the optimal medium, and organisms were tested at both 30 degrees C and 35 degrees C with a standardized inoculum density of 10(3) conidia/mL. No temperature differences were noted in the amount of growth of the dermatophytes tested. With RPMI at an incubation temperature of 35 degrees C, 3 inoculum sizes (10(3), 10(4), and 10(5) conidia/mL) were tested against 4 antifungal agents: griseofulvin, itraconazole, terbinafine, and fluconazole. Inoculum size did not affect minimum inhibitory concentration (MIC) results for itraconazole or terbinafine, but a larger inoculum produced a slightly higher MIC for griseofulvin and a noticeably higher MIC for fluconazole. Our data support the use of RPMI 1640, 35 degrees C, and 4 days as an incubation temperature and time, respectively, and an inoculum of 10(3) conidia/mL as optimal conditions for the determination of the antifungal susceptibility of dermatophytes.
authors.affiliation: Center for Medical Mycology, Mycology Reference Laboratory, University Hospitals of Cleveland and Case Western Reserve University, Ohio 44106, USA.
authors.forename: H A
authors.initials: HA
authors.lastname: Norris
authors.forename: B E
authors.initials: BE
authors.lastname: Elewski
authors.forename: M A
authors.initials: MA
authors.lastname: Ghannoum
mesh_terms: D000935:Antifungal Agents / Q000494:pharmacology*; D003470:Culture Media; D008826:Microbial Sensitivity Tests / Q000379:methods*; D013696:Temperature; D014249:Trichophyton / Q000187:drug effects / Q000254:growth & development*
chemical_list: D000935:Antifungal Agents; D003470:Culture Media