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Photosensitiser Functionalised Nanofiber Fabric for Efficient Light Driven Water Disinfection

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Photosensitiser Functionalised Nanofiber Fabric for Efficient Light Driven Water Disinfection Photosensitiser functionalised nanofiber fabric for efficient light driven water disinfection Hussaini Mohammed Majiya Submitted in accordance with the requirements for the degree of Doctor of Philosophy The University of Leeds School of Biomedical Sciences Faculty of Biological Sciences November, 2017 1 Declaration The candidate confirms that the work submitted is his own, except where work which has formed part of jointly-authored publications has been included. The contribution of the candidate and the other authors to this work has been explicitly indicated below. The candidate confirms that appropriate credit has been given within the thesis where reference has been made to the work of others. Jointly authored publications: (1) Some parts of Chapter One, Chapter Four and Chapter Five have been published in TMPyP functionalised chitosan membrane for efficient sunlight driven water disinfection, 2017, Majiya H, Chowdhury KF, Stonehouse NJ, Millner P.; Journal of Water Process Engineering. Hussaini Majiya contributed all of the research work and writing in this research paper. Kaniz F Chowdhury, Prof. Nicola J Stonehouse and Prof. Paul A Millner helped with research planning, provided critical feedback and editorial support. Copyright © 2017, Elsevier, Journal of Water Process Engineering. (2) Some parts of Chapter One, Chapter Three and Chapter Four have been published in Photodynamic inactivation of bacteriophage MS2: the A- protein is the target of virus inactivation, 2018, Majiya H, Adeyemi O, Stonehouse NJ, Millner P.; Journal of Photochemistry & Photobiology, B: Biology. Hussaini Majiya did all of the research work and most of the writing in this research paper. Dr Oluwapelumi Adeyemi in addition to editorial support, provided PyMOL structures of MS2 capsid and A-protein. Prof. Nicola J Stonehouse and Prof. Paul A Millner helped with research planning, provided critical feedback and editorial 2 support. Copyright © 2017, Elsevier, Journal of Photochemistry & Photobiology, B: Biology. This copy has been supplied on the understanding that it is copyright material and that no quotation from the thesis may be published without proper acknowledgement. © 2017 The University of Leeds and Hussaini Mohammed Majiya The right of Hussaini Mohammed Majiya to be identified as Author of this work has been asserted by him in accordance with the Copyright, Designs and Patents Act 1988. 3 “Valid criticism does you a favour” - Carl Sagan 4 Acknowledgements I appreciate and thankful to Petroleum Technology Development Fund (PTDF), Nigeria for sponsoring my PhD research. I also appreciate my employer- Ibrahim Badamasi Babangida University, Lapai, Nigeria, for giving me a study-leave to pursue my PhD degree. I am grateful to my supervisor, Prof. Paul Millner for his support and educative suggestions throughout my PhD research. My research under supervision of Paul was awesome and I learnt several good things including good supervisor-student relationship. He was always encouraging, ever ready to listen and humour. He never for once talked down my write-ups, suggestions and experiments. These gave me confidence and ability to be a driver of my research and also improved my independent and critical thinking ability. His style of supervision has inspired me to become an academic like him in my future career. I appreciate the efforts of my co-supervisor, Prof. Nicola Stonehouse. She was always there for me especially in virology. I learnt several skills under her supervision. She made me a better scientist and will forever be grateful. I am also grateful to my assessor Dr Alexander O'Neill. His patience, comments and feedback has tremendously improved my work. Thanks to Mr Martin Fuller for his kindness, enthusiasm and expertise in teaching me TEM and doing TEM of most of my samples. Special thanks to all of the previous and present Millner group members I have met during my PhD especially Dr Carolyn Jackson, Dr Asif Ahmed, Dr Jack Goode and Dr Tim Gibson. All of you have made my journey smooth some way or other. I am thankful to Nicola Stonehouse group especially Dr Oluwapelumi Adeyemi and Dr Morgan Herod for their patience and kindness in 5 teaching me many skills in virology. Thanks to Prof. Peter Stockley and group especially Mrs Amy Barker for providing me with the stock samples of bacteriophages MS2 and Qβ and their E. coli host. Thanks to Dr A. M. Afifi for her kind gift of chitosan/polyethylene oxide electrospun nanofiber mats we used in this work. I will forever be grateful to my family members especially my parents for their supports, blessings and prayers. My brothers and sisters especially Hassan were encouraging and supportive. My beloved wife Hauwa Abubakar and loving daughters Ummulkhairri and Fatima have made my every stressful day easier with their love, affection and presence. Finally, to all my friends and well-wishers, you all mean a lot to me, I thank you all. 6 Abstract Sunlight-driven water disinfection system could help provide clean water to some of the world’s poorest regions where contaminated surface water is a major public health problem and bright solar irradiation is available for free. In this work, photosensitiser - 5, 10, 15, 20-tetrakis (1-methyl-4-pyridinio) porphyrin tetra p-toluene sulfonate (TMPyP) was chosen and immobilised onto chitosan nanofiber mats and chitosan membranes for photodynamic disinfection of water since preliminary studies with TMPyP in solution showed it caused a high rate of photodynamic inactivation (PDI) of model viral organisms (bacteriophages MS2 and Qβ, murine norovirus and bovine enterovirus 2). Native gel electrophoresis, SDS-PAGE and western blotting, TEM and DLS were used to analyse pre- and post-PDI samples of the model viruses. The rate of PDI in model viruses was in the order MS2 > phage Qβ > murine norovirus > bovine enterovirus 2. Our data showed that PDI caused aggregation of MS2 particles and crosslinking of MS2 coat protein. However, the aggregation and crosslinking did not correlate to the rate of PDI we observed in MS2. Using sequence specific antibodies raised against MS2 A-protein (host attachment protein), our results suggest that the rate of PDI is relative to loss of antigenicity of sites on the A-protein. The differences in the rate of PDI were compared to amino acid compositions and surface accessibility of host attachment proteins/sites of the model viruses. Possible modes of action are discussed as a means to gaining insight to the targets and mechanisms of PDI of viruses. Chitosan electrospun nanofibers and chitosan membranes were modified by pyromellitic dianhydride in order to introduce carboxyl groups and facilitate adsorption of the cationic TMPyP. The physico-chemical properties of these modified nanofibers and membranes were investigated by microscopy, absorption spectroscopy, Fourier-transform infrared spectroscopy and Midland surface blotting approaches.The chitosan 7 nanofiber/membrane-TMPyP composite showed photodynamic inactivation of MS2 and E. coli BL21. 8 Abbreviations 0PS photosensitizer (ground state) 1PS photosensitizer (singlet excited state) 3PS photosensitizer (triple excited state) BEV 2 Bovine enterovirus 2 Biotin-NHS Biotin-tagged N-Hydroxysuccinimde CM Chitosan membrane CM-T TMPyP functionalised chitosan membrane CPN Chitosan/polyethylene oxide electrospun nanofiber CPN-T TMPyP functionalised Chitosan/polyethylene oxide electrospun nanofiber DBPs Disinfection by-products DMEM Dulbecco's Modified Eagle's Medium ECL Enhanced chemiluminescence EDC 1-Ethyl-3-(3-dymethylaminopropyl) carbodiimide FBS Foetal bovine serum FDA fluorescein diacetate HAAs Haloacetic acids MB Methylene blue MEM Minimum Essential Medium MF Microfiltration MNV Murine norovirus NF Nanofiltration NHS N-Hydroxysuccinimde ester P/S penicillin – streptomycin PDI Photodynamic inactivation PDT Photodynamic therapy PEO polyethylene oxide PI Propidium iodide PMA Pyromelitic dianhydride PS Photosensitiser Q Net charge RB Rose Bengal RO Reverse osmosis ROS Reactive oxygen species TCID50 50 % tissue culture infective dose THMs Trihalomenthanes TMPyP 5, 10, 15, 20-tetrakis (1-methyl-4-pyridinio) porphyrin tetra (p- toluenesulfonate) TSA Tryptic soy agar TSB Tryptic Soy Broth 9 UF Ultrafiltration 10 Table of Contents Declaration ......................................................................................................... 1 Acknowledgements ............................................................................................ 4 Abstract .............................................................................................................. 6 Table of Contents ............................................................................................. 10 Lists of Tables .................................................................................................. 14 Lists of Figures ................................................................................................. 15 Chapter 1: Introduction .................................................................................... 19 1.1 Overview ................................................................................................ 19 1.2 water availability and public health importance ...................................... 21 1.3 Waterborne diseases and the need for alternative water disinfection methods ....................................................................................................... 23 1.4 Wastewater treatment and disinfection .................................................
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