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- Title
Morphological changes and induction of antifungal resistance within expression and mutation the Cyp51 A, B, C genes in Aspergillus fumigatus and Aspergillus flavus due to different CO<sub>2</sub> levels.
- Authors
Darabian, Sima; Rezaie, Sassan; Hashemi, Sayed Jamal; Khodavaisy, Sadegh; Sharifynia, Somayeh
- Abstract
Introduction: Aspergillosis is one of the most common opportunistic fungal diseases in immunocompromised and neutropenic patients. Aspergillus fumigatus is the most common cause of aspergillosis and Aspergillus flavus is the second agent. Due to changes in the concentration of CO2 that some pathogens encounter during the infection process and to understand the role of CO2 as a carbon base. In this study evaluated of changes in antifungal susceptibility patterns, the expression and the mutation in the genes of intervener in Cyp51A,B,C in A.fumigatus and A.flavus in the effect of variable CO2 concentrations. Materials and Methods: A. fumigatus and A.flavus strains were cultured and incubated under the 1%, 3%, 5% and 12% of CO2 concentrations, each time in one, two, and four weeks. The control culture were maintained for 1 week without CO2 concentration. Morphological changes were investigated and antifungal susceptibility tests were performed according to recommendations stated in the Clinical and Laboratory Standards Institute (CLSI) M38-A2 document and evaluated the expression and the mutation in the genes of intervener in Cyp51A,B,C with Real Time PCR protocols. All tests of different CO2 concentrations were compared with control sample. Result: We found that 1%, 3%, 5%, and 12% CO2 concentration were associated with morphological colony changes. Macroscopically, the colonies were shallow dark green, smooth, crisp to powdery with reduced growth; microscopic examination revealed the absence of conidiation. The induction of antifungal resistance of susceptible strains to itraconazole, voriconazole and amphotericin B increased after expose with 12% concentration of CO2 and four weeks of incubation. The MIC value for itraconazole, voriconazole and amphotericin B, were 16μg/ml, 2μg/ml and 16μg/ml, respectively in A.fumigatus and 8μg/ml, 2μg/ml and 16μg/ml in A.flavus. These values for control groups were 0.125 μg/ml, 0.125 μg/ml and 2 μg/ml, respectively in A.fumigatus and 1μg/ml, 0/5μg/ml and 2μg/ml in A.flavus. Also, were not observated significant point mutation in the sequences of gene Cyp51A in A.fumigatus and Cyp51C in A.flavus. The results of study, were showed both increase and decrease in the expression in the genes Cyp51A,B in A.fumigatus and Cyp51C in A.flavus, concomitant with B-actin test, in the compared to control cultures. Conclusion: Exposure to different CO2 concentrations inducted morphological changes and a significant increase the MIC values with increasing expression and without mutation in the Cyp51A,B,C genes of in A.fumigatus and A.flavus, as well. In parallel, resistance to both itraconazole and voriconazole was also observed.
- Subjects
IRAN; ASPERGILLUS; CARBON dioxide; CONFERENCES &; conventions; DRUG resistance in microorganisms; GENE expression; GENETIC mutation; CYTOCHROME P-450
- Publication
Current Medical Mycology, 2018, Vol 4, p106
- ISSN
2423-3439
- Publication type
Article
- DOI
10.18502/cmm.4.S1.2018.180