Scholars' Mine Doctoral Dissertations Student Theses and Dissertations Spring 2014 Biomineralization of carbonates in modern microbial sediments and its implications for CO₂ sequestration Varun Gnanaprian Paul Follow this and additional works at: https://scholarsmine.mst.edu/doctoral_dissertations Part of the Geology Commons, and the Geophysics and Seismology Commons Department: Geosciences and Geological and Petroleum Engineering Recommended Citation Paul, Varun Gnanaprian, "Biomineralization of carbonates in modern microbial sediments and its implications for CO₂ sequestration" (2014). Doctoral Dissertations. 2138. https://scholarsmine.mst.edu/doctoral_dissertations/2138 This thesis is brought to you by Scholars' Mine, a service of the Missouri S&T Library and Learning Resources. This work is protected by U. S. Copyright Law. Unauthorized use including reproduction for redistribution requires the permission of the copyright holder. For more information, please contact [email protected]. BIOMINERALIZATION OF CARBONATES IN MODERN MICROBIAL SEDIMENTS AND ITS IMPLICATIONS FOR CO2 SEQUESTRATION by VARUN GNANAPRIAN PAUL A DISSERTATION Presented to the Faculty of the Graduate School of the MISSOURI UNIVERSITY OF SCIENCE AND TECHNOLOGY In Partial Fulfillment of the Requirements for the Degree DOCTOR OF PHILOSOPHY in GEOLOGY AND GEOPHYSICS 2014 Approved by: David J. Wronkiewicz, Advisor Melanie R. Mormile, Co-Advisor Francisca Oboh-Ikuenobe Wan Yang Jamie S. Foster © 2014 Varun Gnanaprian Paul All Rights Reserved iii PUBLICATION DISSERTATION OPTION This dissertation is organized into four main sections. Section 1 (pages 1 to 17) introduces the two main research projects undertaken and states the hypotheses and objectives. Section 2 (pages 18 to 35) describes the sites, materials used and the experimental methodology that has been employed in all the projects. Paper I (pages 36 to 112) covering the first research project, focuses on the investigations on mineralization of carbonates influenced by sulfate-reducing bacteria, and its implications for subsurface carbon dioxide sequestration. Paper I is written in the form of a manuscript to be submitted to Geochimica et Cosmochimica Acta journal. Paper II (pages 113 to 180) describes the second research project, involving characterization of water and microbialites from Storr’s Lake, and is written in the form of a manuscript for submission to the Geobiology Journal. Section 3 (page 181) includes the general conclusions that were obtained from the overall research projects. Appendices A through E have been added for purposes of normal dissertation writing. iv ABSTRACT Several bacterial species influence carbonate mineral precipitation by modifying pH, alkalinity, Ca2+ activity and by providing nucleation sites for mineralization. Two studies have been undertaken to explore microbial influence in the mineralization of carbonates. In the first study, the ability of sulfate-reducing bacteria (SRB) to induce carbonate mineralization was investigated as a means to enhance mineral sequestration of CO2. Sulfate-reducing bacteria enriched from hypersaline Storr’s Lake, The Bahamas; Lake Estancia, New Mexico; and Great Salt Plains Lake, Oklahoma, were tested in reactors under varying CO2 concentrations. Carbonate mineral precipitation was achieved only in reactors with Lake Estancia SRB community and under a pCO2 of < 20 psi. Hydrogen, lactate and formate served as electron donors for SRB. Carbon isotopic studies confirmed that carbon in the carbonate minerals was derived from electron donors, CO2 or bicarbonate ions in the solution. Sulfate-reducing bacteria’s ability to induce immobile carbonate mineralization can be potentially applied to enhance long-term storage of CO2. Secondly, microbially lithified, organo-sedimentary structures called microbialites in the hypersaline Storr’s Lake were investigated to determine the influence of biotic and abiotic components of the lake on the microbialite formation. The lake water revealed fluctuations in several parameters depending upon rainfall and evaporation. Aragonite and Mg-calcite constituted the carbonate mineralogy of the five microbialites morphologic types examined. Microbial diversity studies by 16S rRNA gene analysis revealed high population percentages of anaerobic phototrophs, halo-respirers and sulfate-reducing bacteria and low population of cyanobacteria (3%). The fluctuating water characteristics, varied mineralogy and the low apparent abundance of cyanobacteria, makes Storr’s Lake, a distinct environment to study microbial interaction with their surroundings during carbonate mineralization. v ACKNOWLEDGEMENTS I extend my most sincere and heart-felt appreciation to my advisors, Drs. David Wronkiewicz and Melanie Mormile, for their continued support and guidance, and for pushing me to achieve my goals. I thank Drs. Francisca Oboh-Ikuenobe, Wan Yang and Jamie Foster, my committee members, for their invaluable inputs and direction. I thank the Department of Energy – National Energy Technology Lab under agreement DE-FE0002416, the sponsors for my research. I thank the Donald Radcliffe Trust (Missouri S&T Geology & Geophysics) for partially covering travel expenses and for stipend and partial tuition expenses during Fall 2013. I thank the faculty and staff of the Departments of Geological Sciences & Engineering, and Biological Sciences. Cheers to Krista, Bobby and Alsedik for their help and for providing necessary comedic relief. I thank the staff of the Gerace Research Centre, San Salvador Island, for logistics and assistance in the Bahamas, and the Bahamas Environment Science & Technology Commission (BEST) for permission to export microbialite samples. I also thank Clarissa Wisner, Advanced Material Characterization Laboratory; Eric Bohannan, Materials Research Center; Shreya Gosh; and Honglan Shi, Environmental Research Center, for help with SEM, XRD, pyrosequencing data interpretation and ICP-OES, respectively. I am especially thankful to Dr. Ken MacLeod, Dr. Cheryl Kelley and Shannon Haynes of Mizzou’s Biogeochemistry IRMS Lab for analysis of the isotopic data. I thank Carlos and Josh for help with microscopy and XRD sample preparation. I am grateful to Dr. Emitt Witt, Dr. Eric (Florida), Dr. Wronk and the students in the field trips for their help during sample collection, and for holding my back under water. Finally, no words can express my love and gratitude to my parents, my brother, family and friends for believing in me and encouraging me to pursue my dreams. vi TABLE OF CONTENTS Page PUBLICATION DISSERTATION OPTION ................................................................... iii ABSTRACT ....................................................................................................................... iv ACKNOWLEDGEMENTS.. ...............................................................................................v LIST OF ILLUSTRATIONS ............................................................................................. xi LIST OF TABLES ........................................................................................................... xiii SECTION 1. INTRODUCTION ...........................................................................................................1 1.1. GEOLOGICAL CARBON SEQUESTRATION .....................................................1 1.2. MICROBIAL ROLE IN CARBONATE MINERAL PRECIPITATION ...............3 1.3. MICROBIALITES AND CARBONATE MINERALIZATION ...........................10 1.4. HYPOTHESES AND OBJECTIVES.. ..................................................................15 2. EXPERIMENTAL METHODS.....................................................................................18 2.1. STUDY SITES .......................................................................................................18 2.2. WATER ANALYSIS FROM DIFFERENT LAKES ............................................19 2.2.1. In-Situ Water Analysis .................................................................................19 2.2.2. Water Analysis in the Laboratory ................................................................21 2.3. SATURATION INDEX BY USING PHREEQ PROGRAM ................................23 2.4. MINERALOGY, ELEMENTAL ANALYSIS AND MICROSCOPY OF THE SEDIMENTS/MICROBIALITES..........................................................................23 2.4.1. X-Ray Diffraction (XRD) Analysis .............................................................23 2.4.2. Elemental Analysis of the Microbialite Samples .........................................24 2.4.3. Optical and Scanning Electron Microscopic (SEM) Analysis .....................25 2.5. MICROBIAL ENRICHMENT CULTURES .........................................................26 2.6. DNA EXTRACTION AND 16S rRNA GENE ANALYSIS .................................27 2.7. REACTOR TESTING ............................................................................................28 2.7.1. Continuous Reactors ....................................................................................28 2.7.2. Batch Reactors .............................................................................................29 vii 2.8. CARBON ISOTOPE ANALYSIS .........................................................................31 PAPER I. IMPACT OF ELEVATED CO2 CONCENTRATIONS ON THE CARBONATE MINERAL PRECIPITATION ABILITY OF SULFATE-REDUCING BACTERIA AND ITS IMPLICATION FOR CO2 SEQUESTRATION……….…………………..36 ABSTRACT……………………………………………………………...…..…………..36