Microscopic Evaluation, Molecular Identification, Antifungal Susceptibility, and Clinical Outcomes inFusarium,Aspergillusand, Dematiaceous Keratitis

Abstract
Purpose.Fusarium,Aspergillus, and Dematiaceous are the most common fungal species causing keratitis in tropical countries. Herein we report a prospective study on fungal keratitis caused by these three fungal species.Methodology. A prospective investigation was undertaken to evaluate eyes with presumed fungal keratitis. All the fungal isolates (n=73) obtained from keratitis infections were identified using morphological and microscopic characters. Molecular identification using sequencing of the ITS region and antifungal susceptibility tests using microdilution method were done. The final clinical outcome was evaluated in terms of the time taken for resolution of keratitis and the final visual outcome. The results were analyzed after segregating the cases into three groups, namely,Fusarium,Aspergillus, and Dematiaceous keratitis.Results. Diagnosis of fungal keratitis was established in 73 (35.9%) cases out of 208 cases. The spectra of fungi isolated wereFusariumspp. (26.6%),Aspergillusspp. (21.6%), and Dematiaceous fungi (11.6%). The sequence of the ITS region could identify theFusariumandAspergillusspecies at the species complex level, and the Dematiaceous isolates were accurately identified. Using antifungal agents such as fluconazole, natamycin, amphotericin B, and itraconazole, the minimum inhibitory concentrations (MICs) forFusariumspp. were >32 μg/mL, 4–8 μg/mL, 0.5–1 μg/mL, and >32 μg/mL, respectively. Antifungal susceptibility data showed thatCurvulariaspp. was highly resistant to all the antifungal agents. Overall, natamycin and amphotericin B were found to be the most effective antifungal agents. The comparative clinical outcomes in all cases showed that the healing response in terms of visual acuity of the Dematiaceous group was significantly good when compared with theFusariumandAspergillusgroups (P<0.05). The time required for healing in theFusariumgroup was statistically significantly less when compared with theAspergillusand Dematiaceous groups.Conclusion. This study demonstrates important differences in microscopic features of scraping material and antifungal susceptibility between the three groups. Early and accurate identification coupled with the MIC data, and thereby appropriate treatment is crucial for complete recovery.

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