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1 Deoxygenation Treatment Strategy Deoxygenation Treatment Strategy to Control Vampirovibrio chlorellavorus in Chlorella sorokiniana Cultures Item Type text; Electronic Dissertation Authors Attalah, Said Publisher The University of Arizona. Rights Copyright © is held by the author. Digital access to this material is made possible by the University Libraries, University of Arizona. Further transmission, reproduction, presentation (such as public display or performance) of protected items is prohibited except with permission of the author. Download date 02/10/2021 21:48:38 Link to Item http://hdl.handle.net/10150/631467 1 DEOXYGENATION TREATMENT STRATEGY TO CONTROL VAMPIROVIBRIO CHLORELLAVORUS IN CHLORELLA SOROKINIANA CULTURES by Said Attalah __________________________ Copyright © Said Attalah 2018 A Dissertation Submitted to the Faculty of the DEPARTMENT OF BIOSYSTEMS ENGINEERING In Partial Fulfillment of the Requirements For the Degree of DOCTOR OF PHILOSOPHY In the Graduate College THE UNIVERSITY OF ARIZONA 2018 2 3 STATEMENT BY AUTHOR This dissertation has been submitted in partial fulfillment of the requirements for an advanced degree at the University of Arizona and is deposited in the University Library to be made available to borrowers under rules of the Library. Brief quotations from this dissertation are allowable without special permission, provided that an accurate acknowledgement of the source is made. Requests for permission for extended quotation from or reproduction of this manuscript in whole or in part may be granted by the copyright holder. SIGNED: Said Attalah 4 ACKNOWLEDGEMENTS First, I would like to express my highest regards and sincere gratitude to my advisor Dr. Peter Waller for his valuable guidance and continuous support during the course of this research. Without his guidance and dedication, I would not be able to achieve my PhD program. I am greatly thankful to my co-advisor Dr. Kevin Fitzsimmons for his support and advice. I was fortunate to take his classes and learn the essentials about algae. I express my profound gratitude to Dr. Kim Ogden for her advice and help to define and develop my research plan. Her insightful thoughts and pertinent comments were very valuable to my research project. I am honored to have her in my dissertation committee. My deep gratitude goes to Dr. Stephen Poe who accepted to be in my dissertation committee. His advice and encouragement helped me to be persistent and stay focused on my research goals. My sincere acknowledgements are extended to Dr. Judith Brown, who helped me to develop my experimental design and allowed me to use her laboratory equipment. Her insightful comments and suggestions were crucial in the completion of this research. I would like to thank S. Steichen, C. Brown, and C. N. Galvez who provided the stock culture and inoculum and helped with qPCR measurements. I would like to thank S. Gao for his help with nutrients preparation and ash free dry weight measurement. My acknowledgements go also to Dr. M. Kacira, Dr. G. Ogden, Dr. F. Jia, G. Khawam, O. Bertelsen, E. Leichtenberg, K. Lepley, I. Liang, M. Acedo, and Y. Mahdipoor, members of the RAFT team at the University of Arizona. 5 My exceptional thanks go to the BE department faculty and staff members for their continuous support. I am honored and proud to be a part of this great department. I am grateful for the U.S. Department of Energy and Regional Algal Feedstock Testbed (RAFT) project, University of Arizona, for supporting this study. I am so thankful to my family for their love and support that gave me lot of courage and patience to achieve this project. Thank you! 6 TABLE OF CONTENT LIST OF FIGURES .....................................................................................................................11 LIST OF TABLES .......................................................................................................................16 ABSTRACT ..................................................................................................................................17 Chapter 1: Introduction .............................................................................................................19 1.1. Literature review .................................................................................................................19 1.1.1. Microalgae cultivation and biological contaminants issue ...........................................19 1.1.2. Bacteria-algae interaction ............................................................................................22 1.1.3. Chlorella sorokiniana characteristics ..........................................................................23 1.1.4. Vampirovibrio chlorellavorus phenotypes ..................................................................24 1.1.5. Control strategies of biological contaminants .............................................................25 1.2. Rationale ..............................................................................................................................26 1.3. Hypotheses ..........................................................................................................................27 1.3.1. Hypothesis 1 .................................................................................................................27 1.3.2. Hypothesis 2 ................................................................................................................27 1.4. Dissertation structure ...........................................................................................................28 Chapter 2: Deoxygenation in algae cultivation systems ..........................................................31 2.1. Introduction .........................................................................................................................31 7 2.2. Dissolved oxygen measurement methods ...........................................................................31 2.3. Deoxygenation approaches .................................................................................................32 2.4. Deoxygenation impact on microalgae and associated microorganisms .............................33 Chapter 3: Technical evaluation and cost estimation of the deoxygenation process ............36 3.1. Introduction ........................................................................................................................36 3.2. Dissolved oxygen removal .................................................................................................37 3.3. Deoxygenation with nitrogen gas sparging ........................................................................38 3.4. Technical considerations and cost estimation of the deoxygenation process ....................38 Conclusion ....................................................................................................................................40 Complete Dissertation References ..............................................................................................41 Appendix A: Application of deoxygenation-aeration cycling to control the predatory bacterium Vampirovibrio chlorellavorus in Chlorella sorokiniana cultures ...........................55 Abstract .....................................................................................................................................55 1. Introduction ...........................................................................................................................56 2. Materials and methods ...........................................................................................................60 2.1. Pathogen-free cultures and co-cultures ..........................................................................60 2.2. Deoxygenation-aeration cycling experiments ................................................................61 2.3. Inoculum preparation and experimental media ..............................................................63 2.4. Algal suspension culture growth assessment .................................................................63 8 2.5. Real time quantitative polymerase chain reaction for quantification of V. chlorellavorus ................................................................................................................................................65 2.6. Experimental reactor design and laboratory apparatus ..................................................65 3. Results and discussion ............................................................................................................68 3.1. Effect of deoxygenation cycling on V. chlorellavorus -free C. sorokiniana ...................68 3.2. Impact of deoxygenation-aeration cycling on V. chlorellavorus infection ....................74 4. Conclusion ..............................................................................................................................83 5. Acknowledgements ...............................................................................................................83 6. References ..............................................................................................................................83 Appendix B: Technoeconomic assessment of deoxygenation for control of Vampirovibrio chlorellavorus in Chlorella sorokiniana cultures .......................................................................92 Abstract .....................................................................................................................................92
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