University of Windsor Scholarship at UWindsor Electronic Theses and Dissertations 2014 Biological hydrogen production from lignocellulosic biomass in an up-flow anaerobic sludge blanket reactor using mixed microbial cultures Sathyanarayanan Sevilimedu Veeravalli University of Windsor Follow this and additional works at: http://scholar.uwindsor.ca/etd Recommended Citation Sevilimedu Veeravalli, Sathyanarayanan, "Biological hydrogen production from lignocellulosic biomass in an up-flow anaerobic sludge blanket reactor using mixed microbial cultures" (2014). Electronic Theses and Dissertations. Paper 5088. 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). 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BIOLOGICAL HYDROGEN PRODUCTION FROM LIGNOCELLULOSIC BIOMASS IN AN UP-FLOW ANAEROBIC SLUDGE BLANKET REACTOR USING MIXED MICROBIAL CULTURES By Sathyanarayanan Sevilimedu Veeravalli 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 Sathyanarayanan Sevilimedu Veeravalli Biological Hydrogen Production From Lignocellulosic Biomass in an Up-flow Anaerobic Sludge Blanket Reactor using Mixed Microbial Cultures by Sathyanarayanan Sevilimedu Veeravalli APPROVED BY: ______________________________________________ B. Liao, External Examiner Lakehead University ______________________________________________ D. Heath The Great Lakes Institute for Environmental Research (GLIER) ______________________________________________ R. Seth Department of Civil & Environmental Engineering ______________________________________________ X. Xu Department of Civil & Environmental Engineering ____________________________________________ J. Lalman, Advisor Department of Civil and Environmental Engineering 8 May 2014 Declaration of Co-Authorship / 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 D. Heath. The contribution of co-authors is limited to the provision of microbiological results which are included in Chapters 4, 5, 6.2, 6.3, 7 and 8 of the dissertation. In all cases, the experimental design was sent to Dr. Jerald A. Lalman by the author, based on his suggestion and recommendations made, the 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 qualifications, this dissertation, and the research to which it refers, is the product of my own work. II. Previous Publication: Chapter 6: This chapter was published under title ‘Effect of furans and linoleic acid on hydrogen production’ in Int J Hydrogen Energy , 2013, Volume 28, Issue 18, 12283-1293 (http://dx.doi.org/10.1016/j.ijhydene.2013.07.035 ). The initial draft was written by Mr. Veeravalli, for which Dr. Lalman provided significant changes for publication. The additional recommendations and suggestions given by Dr. Lalman were incorporated in the dissertation Chapter. I would like to thank Dr. Lalman, for his contribution. Chapter 7: This chapter was published under title ‘Optimizing hydrogen production from a switchgrass steam exploded liquor using a mixed anaerobic culture in an upflow iii anaerobic sludge blanket reactor’ in Int J Hydrogen Energy , 2014,Volume 39, Issue 7, 3160-3175 (http://dx.doi.org/10.1016/j.ijhydene.2013.12.057). The initial draft was written by Mr.Veeravalli, which was reviewed by Dr. Lalman and Dr. Chaganti who provided additional recommendations for the improvement. Dr. Lalman provided significant changes for the publication, the additional recommendations and suggestions given by Dr. Lalman were incorporated in the dissertation Chapter. I would like to thank Dr. Lalman and Dr. Chaganti, for their contribution. Chapter 8: This chapter was published under title ‘Fermentative H 2 production from a switchgrass steam exploded liquor fed to mixed anaerobic cultures: Effect of hydraulic retention time, linoleic acid and nitrogen sparging’ in Int J Hydrogen Energy , 2014 (In Press) (http://dx.doi.org/10.1016/j.ijhydene.2014.04.114). The initial draft was written by Mr. Veeravalli, which was reviewed by Dr. Lalman who provided additional recommendations for the improvement. Dr. Lalman provided significant changes for the publication, the additional recommendations and suggestions given by Dr. Lalman were incorporated in the dissertation Chapter. I would like to thank Dr. Lalman, for his contributions. I hereby certify that no other part of this dissertation has been published or submitted for publication. 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 current research investigated hydrogen (H 2) production potential from lignocellulosic biomass via dark-fermentation in upflow sludge blanket reactors (UASBRs) using mixed anaerobic culture. The effects of hydraulic retention time (HRT) and organic loading rate (OLR), on H 2 production were examined under mesophilic conditions using linoleic acid (LA), as a methanogenic inhibitor. The dynamics of the microbial community were explored using terminal restriction fragment length polymorphism analysis. -1 Studies with pure glucose revealed that high H2 yield ≥ 2.1 mol mol glucose was obtained in control cultures operating at HRTs ranging from 12 h to 20 h with OLRs corresponding to 16 g L -1 d-1 and 10 g L -1 d-1, respectively. Species belonging to Clostridia was observed under these conditions. A further decrease with the HRT in -1 control cultures reduced H 2 yields up to 1.3 mol mol glucose, while addition of LA -1 showed improved H 2 yields ≥ 2.0 mol mol glucose at HRTs ranging from 6 to 12 h. -1 A maximum H 2 yield of 303±20 mL g COD was obtained from switchgrass-derived sugars under the optimal conditions (pH 5.0, HRT 10 h and 1.75 g L -1 of LA) determined using response surface methodology. The microbial characterization under optimal conditions showed dominance of Ruminococcaceae and Clostridiaceae with efficient suppression of methanogens. Nitrogen sparging of the UASBRs under the optimal conditions, increased H 2 yield by 15% in comparison to unsparged cultures. Sparging the bioreactors increased the abundance of Clostridium sp. and Bacillus sp. under LA treated conditions. -1 A stable H 2 yield of 274±40 mL g COD was obtained by the control cultures fed corn stover hydrolysate and operating at 18 and 24 g COD L -1 d-1, suggesting furans and phenols could serve as methanogenic inhibitors at low levels. The dominance of Clostridium sp., Flavobacterium sp. and Eubacterium sp., were observed under these H2- producing conditions. v The results from current research suggest that H 2 production from lignocellulosic biomass is feasible and could be applied on a large scale by maintaining proper operational conditions. vi DEDICATION I dedicate this dissertation work to my mother Geetha, other family members and friends. vii ACKNOWLEDGEMENTS First I would like to thank the ALMIGHTY, for imparting moral support towards the completion of my doctoral work at Windsor, Ontario, Canada. With a deep sense of gratitude, I wish to express my sincere thanks to my guide Dr. Jerald A. Lalman for his invaluable guidance at every stage of this research project and helping me get out the publications. I would like to acknowledge my committee member Dr. Rajesh Seth , Dr. Iris Xu and Dr. Daniel Heath for reviewing this dissertation and for their valuable comments and suggestion. In particular, I express my sincere thanks to Dr. Daniel D. Heath , Director, Great Lakes Institute of Environment and Research (GLIER), University of Windsor, for funding and providing the lab space in performing the genomic work in his Lab and