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Thiobacillus Denitrificans Nitrate-Dependent, Neutral pH Bioleaching of Ni from an Ultramafic Concentrate by Han Zhou A thesis submitted in conformity with the requirements for the degree of Master of Applied Science Chemical Engineering and Applied Chemistry University of Toronto © Copyright by Han Zhou 2014 ii Nitrate-Dependent, Neutral pH Bioleaching of Ni from an Ultramafic Concentrate Han Zhou Master of Applied Science Chemical Engineering and Applied Chemistry University of Toronto 2014 Abstract This study explores the possibility of utilizing bioleaching techniques for nickel extraction from a mixed sulfide ore deposit with high magnesium content. Due to the ultramafic nature of this material, well-studied bioleaching technologies, which rely on acidophilic bacteria, will lead to undesirable processing conditions. This is the first work that incorporates nitrate-dependent bacteria under pH 6.5 environments for bioleaching of base metals. Experiments with both defined bacterial strains and indigenous mixed bacterial cultures were conducted with nitrate as the electron acceptor and sulfide minerals as electron donors in a series of microcosm studies. Nitrate consumption, sulfate production, and Ni released into the aqueous phase were used to track the extent of oxidative sulfide mineral dissolution; taxonomic identification of the mixed culture community was performed using 16S rRNA gene sequencing. Nitrate-dependent microcosms that contained indigenous sulfur- and/or iron-oxidizing microorganisms were cultured, characterized, and provided a proof-of-concept basis for further bioleaching studies. iii Acknowledgments I would like to extend my most sincere gratitude toward both of my supervisors Dr. Vladimiros Papangelakis and Dr. Elizabeth Edwards. This work could not have been completed without your brilliant and patient guidance. Your enthusiasm and curiosity in scientific pursuit and teaching is a constant source of motivation in my work; your energy and optimistic take on life will forever guide me and I will always look upon you two as role models in life. NSERC and our industrial sponsor Vale deserve recognition for providing funding and the ore material for my research. I would like to thank Susie (Endang Susilawati) and Line Lomheim in Biozone in teaching me how to work in a large microbiology lab on a daily basis and always being there to offer their help with lab related issues and where to find stuff. (It’s surprising how big a difference it makes!) I would also like to recognize Paul Jowlabar for his help in earlier stages of my project with building my first set of reactors, and George Kretschemann for his expertise in mineralogy analysis. I want to thank all my lab mates and colleagues, Georgiana, Doug, Srinath, Fei, Cheryl, Shuiquan, Chris, and everyone else in both APEC lab and Edlab, your friendship, guidance, and generosity have made the past two and a half years a memorable and rewarding experience in my life. The most sincere gratitude and love goes to my parents, Bing Zhou and Jiang Gao, for their unconditional love and support in my education and all other aspects in life. Without them, I could not have become the person I am today. Finally, I would like to dedicate this work to my grandfather, Ruju Zhou. iv Table of Contents Acknowledgments ............................................................................................................................... iii Table of Contents ................................................................................................................................. iv List of Tables ......................................................................................................................................... vii List of Figures ........................................................................................................................................ ix List of Appendices ................................................................................................................................ xi 1 Introduction ..................................................................................................................................... 1 1.1 Overview ................................................................................................................................................. 1 1.2 Objectives ............................................................................................................................................... 2 1.3 Outline of Thesis ................................................................................................................................... 3 2 The Thompson Concentrate ....................................................................................................... 4 2.1 Thompson Deposit History ............................................................................................................... 4 2.2 Mineralogy ............................................................................................................................................. 4 2.3 Processing Concerns ........................................................................................................................... 9 2.4 Incentives for the current study ................................................................................................... 10 3 Literature Review on Biohydrometallurgy ......................................................................... 11 3.1 Introduction ......................................................................................................................................... 11 3.2 Acid Bioleaching ................................................................................................................................. 11 3.3 Bioleaching at Elevated pH ............................................................................................................. 12 3.4 Nitrate-dependent Bioleaching System ...................................................................................... 13 3.4.1 Thiobacillus denitrificans ............................................................................................................................ 13 3.4.2 S oxidation by T. denitrificans ................................................................................................................... 14 3.4.3 Fe oxidation by T. denitrificans ................................................................................................................ 14 3.4.4 Nitrite reduction inhibition in T. denitrificans. ................................................................................. 15 3.4.5 Attachment to solid surface ....................................................................................................................... 15 3.4.6 Other nitrate-dependent iron- and sulfur-oxidizing bacteria .................................................... 15 4 Methodology .................................................................................................................................. 17 4.1 Ultramafic concentrate .................................................................................................................... 17 4.1.1 Material Preparation .................................................................................................................................... 17 4.1.2 UMFC Preservation ....................................................................................................................................... 17 v 4.1.3 Acid pre-leach .................................................................................................................................................. 18 4.2 Analytical techniques ....................................................................................................................... 18 4.2.1 Solid material characterization ................................................................................................................ 18 4.2.2 Aqueous Samples ........................................................................................................................................... 20 4.3 Experiments with Defined Culture – Thiobacillus denitrificans (ATCC 25259) ............ 21 4.3.1 Revitalization, Growth, and Maintenance ........................................................................................... 21 4.3.2 Experimental Setup ....................................................................................................................................... 22 4.3.3 Sample Analysis .............................................................................................................................................. 23 4.4 Experiments with Indigenous Cultures – First Enrichment ................................................ 23 4.4.1 Collection of Indigenous Sediment Samples from Thompson .................................................... 24 4.4.2 First Enrichment – Experimental Design ............................................................................................ 24 4.4.3 Microcosm Setup ............................................................................................................................................ 25 4.4.4 Sample Analysis .............................................................................................................................................. 25 4.5 Second Enrichment ...........................................................................................................................
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