Phase-Dependent Differential In Vitro and Ex Vivo Susceptibility of <i>Aspergillus flavus</i> and <i>Fusarium keratoplasticum</i> to Azole Antifungals
Darby Roberts | Jacklyn Salmon | Marc A. Cubeta | Brian C. Gilger
Fungal keratitis (FK) is an invasive infection of the cornea primarily associated with <i>Aspergillus</i> and <i>Fusarium</i> species. FK is treated empirically with a limited selection of topical antifungals with varying levels of success. Though clinical infections are typically characterized by a dense network of mature mycelium, traditional models used to test antifungal susceptibility of FK isolates exclusively evaluate susceptibility in fungal cultures derived from asexual spores known as conidia. The purpose of this study was to characterize differences in fungal response when topical antifungal treatment is initiated at progressive phases of fungal development. We compared the efficacy of voriconazole and luliconazole against in vitro cultures of <i>A. flavus</i> and <i>F. keratoplasticum</i> at 0, 24, and 48 h of fungal development. A porcine cadaver corneal model was used to compare antifungal efficacy of voriconazole and luliconazole in ex vivo tissue cultures of <i>A. flavus</i> and <i>F. keratoplasticum</i> at 0, 24, and 48 h of fungal development. Our results demonstrate phase-dependent susceptibility of both <i>A. flavus</i> and <i>F. keratoplasticum</i> to both azoles in vitro as well as ex vivo. We conclude that traditional antifungal susceptibility testing with conidial suspensions does not correlate with fungal susceptibility in cultures of a more advanced developmental phase. A revised method of antifungal susceptibility testing that evaluates hyphal susceptibility may better predict fungal response in the clinical setting where treatment is often delayed until days after the initial insult.
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