Collagenase was found to be the most important enzyme, produced by mycotic keratitis fungi. Therefore, Aspergillus flavus collagenase enzyme has been purified by ammonium sulphate precipitation, Sephadex G25 and DEAE-cellulose chromatography. Electrophoretic analysis for the purified enzyme indicated one subunit of molecular weight of 70-90 KDa when examining on SDS-PAGE. Cetrimide (cetyl trimethyl ammonium bromide) has been tested against the purified collagenase enzyme and indicated reversible competitive inhibitor (kis = 0.15 mg/ml) with high promising activity. Cetrimide might be used to inhibit mycotic keratitis fungi.
IN THE present study, Nickel oxide (NiO) nanoparticles (NiONPs) were bio-synthesized extracellularly by the fungus Fusarium verticillioides (F. verticillioides) using Nickel Nitrate Hexahydrate Ni(NO 3 ) 2 .6H 2 O as a starting material. The formed NiONPs were precipitated in the form of a dark precipitate after adding the fungal filtrate to Ni(NO 3 ) 2 .6H 2 O solution. The properties of this formed precipitate were characterized by X-ray diffraction (XRD), transmission electron microscopy (TEM) and Fourier transformed infrared (FTIR). The XRD pattern indicated that the NiONPs had a face-centered cubic (FCC) structure and their average crystallite size was found to be 8-17.7 nm. The average particle size found to be 8.5-20.7 nm from TEM image, which was compatible with XRD result. The FTIR analysis bands revealed the chemical composition bonding of the formed nano-NiO compound.The antifungal activity of the formed NiONPs was tested against some human pathogenic fungi which were isolated from mycotic keratitis patients, inpatient department of ophthalmology hospital, Tanta University, Egypt. The best results obtained against Candida albicans, C. tropicalis and Aspergillus niger where the minimum inhibitory concentration (MIC) was 12.5, 25 and 25 mg/ml, respectively. Further confirmation of NiONPs biological activity was employed by measurement of their concentration at different intervals within skin, blood and liver of experimental animals, showing a promising absorption and washing out rates from mice tissues; these results indicated that the NiONPs did not accumulated within mice bodies.
The causative mycotic corneal ulcers Aspergillus flavus has been tested in vitro and in vivo against some antifungal agents. Cetrimide was the highest inhibitory agent and has been recorded inhibition zone of 5.7 cm in vitro at concentration of 20 mg/ml. The histopathological effects of cetrimide application on corneal tissues on volunteer's ulcers revealed no abnormalities on corneal tissue. The current results indicate that cetrimide has the highest antifungal activity with no side effects on treatment of A. flavus corneal ulcers. In conclusion: cetrimide is a promising antifungal agent for treatment of human A. flavus mycotic ulcers.
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