A COMPARATIVE STUDY ON ANTIFUNGAL ACTIVITY OF FE2O3, AND FE3O4 NANOPARTICLES.

Ashraf A. Abd El-Tawab 1 , Ahmed, M. A 2 , ElDiasty, E.M. 3 , Fatma I. El-Hofy 4 and * Ahmed Youssef M. Metwally 5 . 1. Bacteriology, Immunology and Mycology Department, Faculty of Veterinary Medicine, Benha University, Egypt. 2. Materials Science Lab. (1), Physics Department, Faculty of Science, Cairo University, Giza, Egypt. 3. Department of Mycology, Animal Health Research Institute, Dokki, Giza, Egypt. 4. Bacteriology, Immunology and Mycology Department, Faculty of Veterinary Medicine, Benha University, Egypt. 5. Department of Bacteriology, Animal Health Research Institute, Zagazig lab, Sharqia, Egypt. ...................................................................................................................... Manuscript Info Abstract ......................... ........................................................................ Manuscript History

Nanoparticle is a new innovation of using particles in a new size which ranged from1-100 nanometers that equal 1 X10 -6 meter. In this study we evaluated the antifungal effect of Fe 2 O 3

Introduction:-
Nanotechnology has offered great possibilities in various fields of science and technology (Adibkia et al., 2007). Pharmaceutical nanotechnology with numerous advantages has growingly attracted the attention of many researchers. Nanoscale, Size range from approximately from 1 nm to 100 nm. This definition is accompanied by two notes: Note 1: Properties that are not extrapolations from a larger size will typically, but not exclusively, be exhibited in this size range. For such properties the size limits are considered approximate. Note 2: The lower limit in this definition (approximately 1 nm) is introduced to avoid single and small groups of atoms from being designated as nano-objects or elements of nanostructures, which might be implied by the absence of a lower limit (ASTM 2006).
Nanoparticles and nanocomposites prepared chemically by precipitation or in situ formation in a given matrix through the sol-gel processes (Sanchez et al., 1996). The conditions determinants of nanoparticle growth are changed in the dependence on method preparation of nanoparticles. Materials scientists and engineers have made significant developments in the improvement of methods of synthesis of nanomaterial solids (Hansany et al., 2012).

ISSN: 2320-5407
Int. J. Adv. Res. 6(1), 189-194 190 Aspergillus flavus is one of the most important pathogenic fungi that contaminate grains or feed mixtures and cause serious problems in such as mycotoxicosis due to its aflatoxins production (Hassan et al., 2014). Development of new and effective antimicrobial agents seems to be of paramount importance, through the antimicrobial activity of metals having various properties, potencies and spectra of activity, has been known and applied for centuries especially in nano-size (Malarkodi et al., 2014).
The aim of this study is the founding of a new effective antifungal compounds through the nanotechnology against Aspergillus flavus which is the most dangerous fungi on animal productions.
Materials and Methods:-Synthesis of nanoparticles:-Fe 2 O 3 nanoparticles synthesized by co-precipitation technique method by Using titra-hydrate ferrous chloride FeCl2.4H2O (> 99:9%) dissolved in distilled water. The ammonium hydroxide NH4OH were added drop-wise to the mixture with stirring under strong ultrasonic agitation was added to the solution to adjust the pH value at 9 or10 till precipitation occurred. Dark brown precipitate is formed and washed using distilled water for several times to remove excess ammonia (10 times Characterization of synthesized nanoparticles:-X-ray diffraction analyses were carried out to identify the previously prepared nanoparticles in pure single phase. Also it confirms the successful formation of these nanoparticles according to (Cullity et al., 2001) Infrared Spectra were recorded on a Perkin -Elmer (FT-IR): FT-IR were carried out to identify the synthesized nanoparticles and to determine its size according to (Guan et al., 2003). Sterilized loop wire was used to transfer A. flavus to sabouraud dextrose agar and incubated at 25 °C for 5 days. From the A. flavus strain, small portion was transferred to 3ml of sabouraud dextrose broth media separately and incubated at 25°C for 24 hrs.0.1 ml of the above five medias were transferred to five different stoppered conical flasks containing 0.9% NaCl solution.1ml of media was taken in a test tube, to which, 1ml of test solution (100 µg/ml) was added. Thereafter, 0.1ml of the microbial strain (A. flavus) prepared in 0.9% NaCl was added to the test tube containing media and test solution. Serial dilution were done five times giving concentrations of 50, 25, 12.5, 6.25, 3.75, 1.5 µg/ml. The test tube were stoppered with cotton and incubated at 25°C for one week. The MIC values were taken as the lowest concentration of the particles in the test tube that showed no turbidity after incubation. The turbidity of the contents in the test tube was interpreted as visible growth of microorganisms. The minimum fungicidal concentration (MFC) was determined by sub culturing 50µl from each test tube showing no apparent growth. Least concentration of test substance showing no visible growth on sub culturing was taken as MFC.
Scanning electron microscope examination of A. flavus after the exposure to Fe 2 O 3 and Fe 3 O 4 : In conical flasks one liter containing 200 ml of yeast extract broth (YEB) than the flasks were autoclaved for 15 minutes at 121 °C cooled at room temperature. The Fe2O3 and Fe3O4 nanoparticles were taken separately at different concentrations of 1.5, 3, 4.5 and 6 mg/100 ml, each concentration dissolved in sterilized distilled water 191 using an ultrasound bath. Preparation of spore suspensions of A. flavus was standardized to 10 6 spores/ml then adding one millimeter of spore suspension of A. flavus and different concentrations of Fe 2 O 3 and Fe3O4 nanoparticles were add to Yeast extract broth (YEB). All the inoculated flasks were incubated at 25°C for 21 days. After the end of incubation period, the content of each flask was filtrated. The treated fungi mycelia sections were collected, fixed with formaldehyde, washed with phosphate buffer solution and dehydrated with alcohol solution (30, 60, 80, 90 and 100%, maintaining the mycelia at 100%) and then submitted to critical point drying according to (Al-othman et al., 2014). Aspergillus flavus became ready for scanning electron microscopy (SEM) using JEOL (JSM-6380 LA) instrument.

Results:-
Phase identification and structural analysis were carried out by XRD and FTIR spectra at room temperature for Fe 2 O 3 and Fe 3 O 4 nanoparticles showed that the investigated sample crystallized in a single phase 45nm size of Fe 2 O 3 nanoparticles and 9nm size of Fe 3 O 4 nanoparticles as shown in Fig 1, 2, 3