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Impact of Chemical Stressors on Hydrogen Metabolism University of Windsor Scholarship at UWindsor Electronic Theses and Dissertations Theses, Dissertations, and Major Papers 2014 Impact of chemical stressors on hydrogen metabolism SARAVANAN RAMIAH SHANMUGAM University of Windsor Follow this and additional works at: https://scholar.uwindsor.ca/etd Recommended Citation RAMIAH SHANMUGAM, SARAVANAN, "Impact of chemical stressors on hydrogen metabolism" (2014). Electronic Theses and Dissertations. 5094. https://scholar.uwindsor.ca/etd/5094 This online database contains the full-text of PhD dissertations and Masters’ theses of University of Windsor students from 1954 forward. These documents are made available for personal study and research purposes only, in accordance with the Canadian Copyright Act and the Creative Commons license—CC BY-NC-ND (Attribution, Non-Commercial, No Derivative Works). Under this license, works must always be attributed to the copyright holder (original author), cannot be used for any commercial purposes, and may not be altered. Any other use would require the permission of the copyright holder. Students may inquire about withdrawing their dissertation and/or thesis from this database. For additional inquiries, please contact the repository administrator via email ([email protected]) or by telephone at 519-253-3000ext. 3208. IMPACT OF CHEMICAL STRESSORS ON HYDROGEN METABOLISM By Saravanan Ramiah Shanmugam A Dissertation Submitted to the Faculty of Graduate Studies through the Department of Civil and Environmental Engineering in Partial Fulfillment of the Requirements for the Degree of Doctor of Philosophy at the University of Windsor Windsor, Ontario, Canada 2014 © 2014 Saravanan Ramiah Shanmugam Impact of Chemical Stressors on Hydrogen Metabolism by Saravanan Ramiah Shanmugam APPROVED BY: ______________________________________________ A. Lau, External Examiner University of British Columbia ______________________________________________ J. Ciborowski Department of Biological Sciences ______________________________________________ R. Seth Department of Civil & Environmental Engineering ______________________________________________ D. Heath The Great Lakes Institute for Environment Research (GLIER) ______________________________________________ J. Lalman, Advisor Department of Civil & Environmental Engineering May 05, 2014 DECLARATION OF CO-AUTHOURSHIP/ PREVIOUS PUBLICATION I. CO-AUTHORSHIP DECLARATION I hereby declare that this dissertation incorporates material that is the result of joint research: It incorporates the outcome of laboratory work, which was done by Dr. Subba Rao Chaganti under the supervision of Dr. Daniel Heath; the contribution of co-authors is limited to the provision of microbiological results which are included in chapters 4, 5, 6, 7 and 8 of the dissertation. In all cases, the experimental design was proposed by the author and approved by Dr. Jerald A. Lalman. The key ideas, primary contributions, engineering laboratory work, data analysis and interpretation, were performed by the author. I am aware of the University of Windsor Senate Policy on Authorship and I certify that I have properly acknowledged the contribution of other researchers to my dissertation, and have obtained written permission from each of the co-author(s) to include the above material(s) in my dissertation. I certify that, with the above qualification, this dissertation, and the research to which it refers, is the product of my own work. II. Declaration of previous publication Results from chapters 4, 5 and 8 of this dissertation have been submitted for publication in International Journal of Hydrogen Energy (IJHE), respectively. Draft copy of the manuscript for Chapters 4, 5 and 8 was provided by the author to Dr. Jerald A. Lalman and he has reviewed and provided significant changes with the manuscript for submission. I would like to thank Dr. Jerald A. Lalman in this regard. The results from chapter 5 are accepted for publication in IJHE (Volume 39, Issue 1, pages 249-57; DOI: http://dx.doi.org/10.1016/j.ijhydene.2013.10.084 ). Results from chapter 8 are also accepted for publication in IJHE and is currently in press, corrected proof; DOI: (http://dx.doi.org/10.1016/j.ijhydene.2014.04.115 ). No part of this dissertation except chapters 4, 5 and 8 has been published/ submitted for publication. I certify that the above material describes work completed during my registration as graduate student at the University of Windsor. iii I declare that, to the best of my knowledge, my dissertation does not infringe upon anyone’s copyright nor violate any proprietary rights and that any ideas, techniques, quotations, or any other material from the work of other people included in my dissertation, published or otherwise, are fully acknowledged in accordance with the standard referencing practices. I declare that this is a true copy of my dissertation, including any final revisions, as approved by my dissertation committee and the Graduate Studies office, and that this dissertation has not been submitted for a higher degree to any other University or Institution. iv ABSTRACT The effects of chemical stressing agents on H 2 metabolism were evaluated using thermodynamic, biochemical, genomic and statistical methods. The objectives of this study were to examine the role of homoacetogens and hydrogenotrophic methanogens exposed to different stress treatments under various fermentation conditions. Negligible H2 consumption was observed at mesophilic and thermophilic temperature (at pH 4.5) when combined with the addition of 2 g L -1 linoleic acid (LA). Genomic analysis revealed that LA-treated cultures were dominated by Clostridium sp. whereas control cultures were dominated by homoacetogens and methanogens. Lauric acid (LUA), LA, fish oil and furfural affected H2 consumption similarly to BES. The H 2 consumption (%) of the control and chemically treated cultures revealed the following trend: Control > Fish oil = LA = Furfural > BES > LUA. Treatment with different stressing agents also resulted in the formation of diverse fermentation metabolites. The long-term effects of different culture pretreatments under mesophilic condition resulted in higher mean H2 yields compared to the yields from cultures incubated at thermophilic condition (after 5 glucose additions). Hydrogen consumption studies using long term stress treated cultures showed lower consumption at thermophilic temperature than at mesophilic temperature. Uptake hydrogenase activities correlated positively with the H 2 consumption data. Genomic analysis indicated that both methanogens and homoacetogens were present in control cultures, but they were absent from the pretreated cultures. Studies conducted in anaerobic sequencing batch reactors revealed that lowering the HRT from 37.5 h to 7.5 h reduced the methane yield and increased the H 2 yield. Higher H 2 yields were obtained in cultures operated at thermophilic temperature compared to mesophilic and psychrophilic temperature using corn stalk (CS) as substrate. Cultures fed CS liquor showed lower levels of specific methanogenic activities than cultures fed pure sugars. The results from these studies indicate that all the chemical stressing agents investigated were active against H 2 consumers (methanogens in particular). In addition to different stress treatments, proper control of operational parameters such as pH, HRT and v temperature is required to minimize H 2 consumption and maximize H 2 production in dark fermentation process. vi DEDICATION I dedicate this dissertation to my beloved and wonderful family vii ACKNOWLEDGEMENTS First and foremost I would like to thank the almighty for the continuous support. I would like to offer my sincerest gratitude to my advisor, Dr. Jerald A. Lalman, who supported me throughout this research with his patience and valuable guidance. I attribute the success of this work to his incessant enthusiasm and effort. His encouragement helped me to strive for betterment; personally, academically and professionally. It will always be an honour to be counted as one of his disciples. I would also like to extend my heartful gratefulness to my committee members, Dr. Rajesh Seth, Dr. Daniel Heath, Dr. Jan Ciborowski and Dr. Anthony Lau for spending their time to review my work and for providing comments and suggestions to improve this dissertation. I would also like to extend my appreciation to Dr. Daniel Heath for offering the facilities to perform genomic analysis component of this work and Dr. Jan Ciborowski for helping me with the principal component analysis and cluster analysis. I was blessed to have the most awesome group of lab-mates ever during these four years of stay in this University: Sathyanarayanan Sevilimedu Veeravalli, Kannappan Thiagarajan, Justin Philpot, Matthew Kachler, Kiruba Shankar, Robert Ty, Wuden Ayele Shewa, Bandi Boje Gowda, Chranjeevi Telluri, Rajesh Singh, Gunasekar Varadarajan, Mohanraj Sundaresan and Drs. Subba Rao Chaganti, Saady Noori, Sathish Tadikamala, Sanjay Kumar, Dong-Hoon Kim, Saravanan Rengaraj and Chungman Moon. I thank Sathyanarayanan for his kind and encouraging words during my research. Thanks for their friendly support and promoting a workable atmosphere in the lab. I would like to extend my thanks to Dr. Subba Rao Chaganti for spending his time in providing me with the microbial data for my dissertation. I would also like to thank Dr. Elizabeth Munn for the help she provided. My sincere appreciation is due to all the staff members of the Civil and Environmental Engineering Department:
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