Comparison of CYP51A gene expression of Aspergillus fumigatus isolates against fluconazole and nano-fluconazole

Subject Areas : Journal of Comparative Pathobiology

Sarrafha Sarrafha, ¹ , S.J. Hashemi Hazaveh ² , S. Rezaei ³ , M. Bayat, ⁴

1 - Department of Pathobiology, Science and Research Branch, Islamic Azad University, Tehran, Iran
2 - Department of Medical Parasitology and Mycology, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran
3 - Department of Medical Parasitology and Mycology, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran
4 - Department of Pathobiology, Science and Research Branch, Islamic Azad University, Tehran, Iran

Received: 2022-05-21 Accepted : 2022-05-21 Published : 2021-11-22

Keywords: Aspergillus fumigatus, fluconazole, CYP51A gene, Nanoparticle, minimum inhibitory concentrati,

Abstract :

Aspergillosis is an opportunistic fungal infection in animals. The disease is mainly respiratory, but other disease manifestations also occur in poultry. Researchers have shown that, one of the reasons for the increase in drug resistance in Aspergillus species is the overexpression of the CYP51A gene. This study compared CYP51A gene expression of Aspergillus fumigatus isolated from poultry against fluconazole and nano-fluconazole. Fluconazole liposomal-nanoparticles were prepared by the thin layer hydration method. We dissolved 5.12 mg of fluconazole powder in 1 ml of sterile distilled water and 6 ml of chloroform-methanol organic solvent. We added 51 mg of lecithin and 5 mg of cholesterol to it. The size of nanoparticles was 88.9±12.1 nm and the surface charge of these nanoparticles was -20.12±1.88 mv. We also used a Scanning-Electron Microscope to study the structure of nanoparticles. Thirty samples of A. fumigatus were collected from poultry lung nodules. Minimum inhibitory concentration (MIC) was performed by standard Broth Microdilution method according to NCCLS-M38A2 to evaluate the MIC of fluconazole and nano-fluconazole against isolates. We selected two high-resistance isolates to fluconazole and used them to determine the CYP51A gene expression level by real-time PCR. The results showed that nano-fluconazole had a lower MIC than fluconazole and in lower concentrations of the drug inhibited the growth of Aspergillus fumigatus isolates. CYP51A gene expression was increased in fluconazole and nano-fluconazole-treated isolates compared to the untreated state. Conversely, a decrease in CYP51A gene expression was observed in the exposure to nano-drug compared with the normal drug.

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