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Synthesis of Cobalt Aluminum Oxide and Aluminum Oxide Nanoparticles and Evaluation of their
Antimicrobial Activity Against Some Pathogens
By Esraa Khodor El-Hadidi
Thesis
Submitted in Partial Fulfillment of the Requirements for
the Degree of Master of Science in Biology
Department of Biological Sciences
Faculty of Science
2020
Synthesis of Cobalt Aluminum Oxide and Aluminum Oxide Nanoparticles and Evaluation of their
Antimicrobial Activity Against Some Pathogens
By Esraa Khodor El-Hadidi
Submitted in Partial Fulfillment of the Requirements for
the Degree of Master of Science in Biology
Department of Biological Sciences
Faculty of Science
Supervised by Prof. Dr. HodaYusef Prof. Dr. Ramadan Awad Professor of Microbiology Professor of Material Science
Biological Sciences Department Dean of Faculty of Science
Faculty of Science Faculty of Science
Beirut Arab University Beirut Arab University
2020
iv
Abstract
With the increasing concern for human health and quality of life, the uses of
nanoparticles for disinfection become more and more important. There is a special
interest in induced bactericidal effect of Aluminum Oxide (Al2O3), Cobalt Aluminum
oxide (CoAl2O4), and Aluminum doped Zinc Oxide (Zn0.9Al0.1O) nanoparticles in the
area of coated ceramic and paints production for hospitals and health cares to reduce
the spread of nosocomial infection. The current study outlines the synthesis of Al2O3,
CoAl2O4, and Zn0.9Al0.1O Nps nanoparticles by co-precipitation method and
subsequently the antimicrobial potentials of these nanoparticles was evaluated against
several pathogenic bacteria along with two fungi, mostly nosocomial pathogens. The
formation and the characterization of Al2O3, CoAl2O4, and Zn0.9Al0.1O nanoparticles
were confirmed by UV-Vis Spectroscopy, X-Ray Diffraction (XRD), Fourier
Transform Infra-Red Spectroscopy (FTIR), and Transmission Electron Microscopy
(TEM). All data revealed the formation of pure nano-sized Al2O3, CoAl2O4, and
Zn0.9Al0.1O NPs. Among the tested synthesized nanoparticles, aluminum doped zinc
oxide show powerful antimicrobial action on the tested bacteria and fungi except
Aspergillus sp.. The effectiveness of the synthesized nanoparticles (Al2O3, CoAl2O4,
and Zn0.9Al0.1O NPs) showed that the action of CoAl2O4 and Zn0.9Al0.1O NPs against
tested bacteria was determined to be bactericidal. Zn0.9Al0.1O NPs exhibited a
privileged ability to suppress the growth of nosocomial pathogens in culture media
and paints as an application. Transmission electron images clearly illustrated a
remarkable damage in E. coli cell’s structure upon exposure to Zn0.9Al0.1O NPs, the
observations suggest that Zn0.9Al0.1O NPs distorted and damaged bacterial cell wall
and cytoplasmic membrane resulting in leakage of protoplasmic content.
v
Table of Contents Acknowledgments .............................................................................................................. v
Abstract .............................................................................................................................. iv
Table of Contents ............................................................................................................... v
List of Tables ................................................................................................................... viii
List of Figures .................................................................................................................... ix
List of Abbreviations…………………………………………………………………….xi
1. Introduction ................................................................................................................ 1
1.1. Nanoparticles ............................................................................................................ 1
1.2. Metal Oxide Nanoparticles ....................................................................................... 1
1.2.1. An Overview of metal oxide nanoparticles ....................................................... 1
1.2.2. Physical-Chemical Characterization of Metal Oxide Nanoparticles ................. 2
1.2.3. Antimicrobial Activity of Metal Oxide Nanoparticles ...................................... 2
1.2.4. Mechanisms of Metal Nanoparticles on Microbes ............................................ 3
1.2.4.1. Interactions with Phospholipid Bilayer ........................................................... 4
1.2.4.2. Production of Reactive Oxygen Species (ROS) ............................................ 5
1.2.4.3. Binding to Cytosolic Proteins .......................................................................... 5
1.3. Aluminum Oxide, Cobalt Aluminum Oxide and Aluminum Doped Zinc Oxide Nanoparticles ....................................................................................................................... 5
1.3.1. Aluminum Oxide Nanoparticles ........................................................................ 5
1.3.2. Applications of Aluminum Oxide Nanoparticles .............................................. 6
1.3.3. Cobalt Aluminum Oxide Nanoparticles ........................................................... 7
1.3.4. Applications of Cobalt Aluminum Oxide Nanoparticles .................................. 8
1.3.5. Aluminum Doped Zinc Oxide Nanoparticles .................................................... 8
1.3.6. Applications of Aluminum Doped Zinc Oxide Nanoparticles .......................... 9
1.4. Toxicity of Nanoparticles ......................................................................................... 9
1.5. Hospital Acquired Infection .................................................................................... 10
1.5.1. Overview of Health Care Environment ........................................................... 10
1.5.2. Spread of Nosocomial Infections .................................................................... 11
1.5.3. Routes of Nosocomial Infection Transmission ............................................... 12
1.5.4. Types of Nosocomial Infection ....................................................................... 13
1.6. Control of Nosocomial Infection ............................................................................ 14
1.6.1. Infection Control Programs ............................................................................. 14
1.6.2. Antimicrobial Use and Resistance ................................................................... 15
vi
1.6.2.1 . Appropriate Antimicrobial Use.................................................................... 15
1.6.2.2. Antibiotic Resistance ....................................................................................... 15
1.6.3. Use of Biocides and Bacterial Resistance ....................................................... 16
1.6.3.1. Use of Biocides ................................................................................................ 16
1.6.3.2. Bacterial Resistance to Biocides ...................................................................... 17
1.7. Application of Nanoparticles in Hospitals and Health Care Facilities ................... 17
1.8. Aim of Study ........................................................................................................... 18
2. Materials and Methods ............................................................................................ 19
1.1. Materials ................................................................................................................. 19
2.1.1. Microbial Pathogens ........................................................................................ 19
2.1.2. Chemicals and Supplements ............................................................................ 19
2.1.3. Culture Media .................................................................................................. 19
2.1.4. Reagents........................................................................................................... 20
2.2. Methods................................................................................................................... 21
2.2.1. Preparation of Nanoparticles ............................................................................... 21
2.2.1.1. Preparation of Aluminum Oxide (Al2O3) NPs by Co-Precipitation Method ... 21
2.2.1.2. Preparation of Cobalt Aluminum Oxide (CoAl2O4) NPs by Co-Precipitation Method ………………………………………………………………………….…….22
2.2.1.3. Preparation of Aluminum Doped Zinc Oxide (Zn0.9Al0.1O) NPs by Co-Precipitation Method .......................................................................................................... 22
2.2.2. Nanoparticles Characterization ........................................................................... 23
2.2.2.1. X-ray Powder Diffraction (XRD) .................................................................... 23
2.2.2.2. Transmission Electron Microscope (TEM) ..................................................... 24
2.2.2.3. Ultraviolet-Visible Absorption Spectroscopy (UV-vis) .................................. 25
2.2.2.4. Fourier Transform Infrared- Spectroscopy (FTIR) ......................................... 26
2.2.3. Preparation of Culture Media .............................................................................. 27
2.2.4. Stock Culture of Microorganisms ....................................................................... 27
2.2.4.1. Agar Slants ...................................................................................................... 27
2.2.4.2. Glycerol Stock ................................................................................................. 27
2.2.5. Inoculum Preparation and Standardization ......................................................... 27
2.2.5.1. Preparation of Bacterial Inocula ...................................................................... 27
2.2.5.2. Preparation of Fungal Inocula ......................................................................... 28
2.2.6. Antimicrobial Susceptibility Tests ...................................................................... 28
2.2.6.1. Disk Diffusion method .................................................................................... 28
vii
2.2.6.2. Broth Micro Dilution Method.......................................................................... 29
2.2.6.2.1. Determination of Minimum Inhibitory Concentration (MIC) ................... 29
2.2.6.2.2. Determination of Minimum Bactericidal Concentration (MBC) .............. 29
2.2.6.3. Time Kill Study ............................................................................................... 30
2.2.6.4. Transmission Electron Microscope ................................................................. 30
2.2.7. Application of Aluminum Doped Zinc Oxide Nanoparticles in a Paint Sample 31
3. Results and Discussion ............................................................................................. 32
3.1. Characterization of Al2O3, CoAl2O4, and Zn0.9Al0.1O Nanoparticles ..................... 32
3.1.1. X-ray Diffraction (XRD) ................................................................................. 32
3.1.2. Transmission Electron Microscope (TEM) ..................................................... 35
3.1.3. Ultraviolet -Visible Absorption Spectroscopy (UV-vis) ................................. 36
3.1.4. Fourier Transform Infrared Spectroscopy (FTIR) ........................................... 38
3.2. Antimicrobial Susceptibility Tests .......................................................................... 40
3.2.1. Disc Diffusion Method .................................................................................... 40
2.2.2. Broth Micro Dilution Method.......................................................................... 43
3.2.2.1. Determination of Minimum Inhibitory Concentration (MIC) ................... 43
3.2.2.2. Determination of Minimum Bactericidal Concentration (MBC) .............. 45
3.2.3. Time-kill Study ................................................................................................ 45
3.2.3. Transmission Electron Microscope ................................................................. 60
3.3. Application of Aluminum Doped Zinc Oxide NPs in a Paint Sample ................... 62
4. Recommendation ...................................................................................................... 64
5. Summary ................................................................................................................... 65
6. References .......................................................................................................... 67
67
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