Use of Green Nano-Extracts in the Control of Trichophyton Rubrum

Abstract

The study was conducted in the Postgraduate Laboratory in the College of Education for Pure Sciences and in the Postgraduate Laboratory at the College of Applied Medical Sciences at the University of Karbala and in the Center for Manuscript Preservation , Restoration and Care of Researchers at the Imam Hussain Holy Shrine in Kerbala.The study lasted from 10/10/2019 to 15/10/2020 with the aim of converting the alcoholic extract of Conocarpus erectusregistered in the Global Genebank that accession numbers MT444957 in previous study , to a hybrid nano silver compound (AgcaNps) and its effect on some phenotypic and microbiological characteristics of Trichophyton rubrum IQT-No.1 at different concentrations with different concentrations of the antifungalfluconazole. Some tests were performed for the hybrid nanoparticles, which is a Spectrophotometer, Fourier transform infrared (FT-IR), X-ray diffraction (XRD) and a Scanning Electron Microscope (SEM) to verify the nanoparticles from the alcoholic plant extract. Different concentrations (10, 20, 30, 40, 50, 60, 70 and 80mg) were prepared from the alcoholic extract, the(AgcaNps) hybrid nanoparticles, the fluconazole antifungal and the binary between them .
The virulence of the Trichophyton rubrumIQT-No.1was tested by its effectiveness on the production of enzymes Protease, Lipase and Keratinase,during 3, 6 and 9 days incubation periods at 25 ° Cwhich showed an enzyme activity directly proportional to the incubation period through the average colony diameter (mm). The enzymes differed in their ability to analyze the culture medium through the diameter of the halo formed around the fungal colony in the case of protease and Keratinase enzymes, and the appearance of the white precipitate in the case of the production of the Lipase enzyme.The results showed that the hybrid nanoparticles (AgcaNps) inhibited the diameter of Trichophyton rubrum (8.0) mm more than the alcohol extract of Conocarpus erectus. It did not differ significantly from the antifungal fluconazole with the same concentration of 50%. Concerning the dry weight of the fungal colonies, the hybrid nanocomposite AgcaNps was the most inhibiting of Trichophyton rubrum with a colony weight of 0.01 mg at a concentration of 40% and it did not differ significantly from the antifungal fluconazole with the same concentration,whereas, the alcoholic extract of Conocarpus erectus was the least inhibitor with colony weight 0.02 mg at 40% concentration. The results showed the superiority of the green alcohol extracts of silver nanoparticles in inhibiting the growth of the Trichophyton rubrum. The use of the mixture between the hybrid green nano extracts and the antifungal resulted in reducing the dose of the anti-fungal fluconazole are used, because of its negative effects on human cells and all Eukaryotic organisms.
Many microscopic examinations were performed on the studied fungi and showed the effect of different treatments on the fungal. This effect varied between decomposition or deformations in the fungal mycelium, the separation of protoplasm in some areas, their assembly in other areas in addition to the disappearance of microconidia. The deformation of macroconidia showed internal abnormalities that can be observed, separation of the plasma membrane, and there is another effect that could be observed in the formation of Chlamydospores. This study was the first of its kind in the field of extracting active compounds and converting them into nanoparticle extracts in inhibiting the growth and efficacy of dermatophytes and reducing the recommended dose of the antifungal used in the case of confusion between them.