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Bioremediation of Chromate in Alkaline Sediment-Water Systems

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Bioremediation of Chromate in Alkaline Sediment-Water Systems Bioremediation of chromate in alkaline sediment-water systems Robert Andrew Whittleston Submitted in accordance with the requirements for the degree of Doctor of Philosophy The University of Leeds School of Earth and Environment September 2011 This copy has been supplied on the understanding that it is copyright material and that no quotation for the thesis may be published without proper acknowledgement. The right of R. A. Whittleston to be identified as Author of this work has been asserted by him in accordance with the Copyright, Designs and Patents Act 1988. © 2011 The University of Leeds and Robert A. Whittleston. The candidate confirms that the work submitted is his own, except where work which has formed part of jointly-authored publications has been included. The contribution of the candidate and the other authors to this work has been explicitly indicated below. The candidate confirms that appropriate credit has been given within the thesis where reference has been made to the work of others. Paper status and collaborator contributions Due to the breadth of knowledge required to produce high quality academic research, three of the results chapters within this thesis form all or part of jointly-authored work prepared for publication. Therefore, collaborator contributions and submission status of each article are clarified below. Chapter 4 “Chromate reduction in Fe(II)-containing soil affected by hyperalkaline leachate from chromite ore processing residue” (Submitted to Journal of Hazardous Materials in June 2011, in press August 2011). R. A. Whittleston – principal author, site sampling, geochemical analyses, microcosm experiments, reoxidation experiments, XAS data collection and analyses, culturing of iron reducers, DNA extraction, 16S rRNA gene analysis, clone libraries and phylogenetic tree construction; D. I. Stewart – site sampling, extensive manuscript review; R. J. G Mortimer – manuscript review; Z. C. Tilt – site sampling, XRD, XRF and TOC analyses; A. P. Brown – TEM analysis; K. Geraki – XAS data collection guidance; I. T. Burke – site sampling, SEM preparation, guidance on XAS data collection and analysis, extensive manuscript review. Chapter 5 “Effect of microbially induced anoxia on Cr(VI) mobility at a site contaminated with hyperalkaline residue from chromite ore processing” (Submitted to Geomicrobiology Journal April 2010, published January 2011 28, 68-82). R. A. Whittleston – principal author, geochemical analyses, DNA extraction, 16S rRNA gene analyses, clone libraries and phylogenetic tree construction; D. I. Stewart – site sampling, provided guidance for DNA extraction and 16S rRNA gene analysis, extensive manuscript review; R. J. G Mortimer – manuscript review; D. J. Ashley – IC technical guidance; I. T. Burke – site sampling, microcosm experiments, extensive manuscript review. Chapter 6 “Enhancing microbial reduction in hyperalkaline, chromium contaminated sediments by pH amendment” (Submitted to Journal of Hazardous Materials September 2011). R. A. Whittleston – principal author, site sampling, microcosm experiments, geochemical analyses, DNA extraction, 16S rRNA gene analysis, clone libraries, MOTHUR, rarefaction and MDS analyses, phylogenetic tree construction; R. J. G Mortimer – manuscript review; I. T. Burke – site sampling, extensive manuscript review; D. I. Stewart – site sampling, guidance on MOTHUR, rarefaction and MDS analyses, extensive manuscript review. ACKNOWLEDGEMENTS I would like to express my deepest gratitude to Ian Burke, Doug Stewart, and Rob Mortimer for their unwavering support in my academic development. They have provided me with priceless guidance from their vast range of expertise, without which this project would not have been possible. I am also incredibly grateful to them for the personal help and advice they have given me over the last three or more years. Your ability to remain good humoured in the most frustrating situations was invaluable! I would also like to thank the many others who have had input in one form or another, including but not limited to; David Ashley, Andy Brown, Louise Fletcher, Lesley Neve and Eric Condliffe for technical guidance and data collection, Michael Marsden, Zana Tilt and James Atkins for help with sample collection and interpretation. Thanks also to Kalotina Geraki and Fred Mosselmans for their help with XAS data collection and interpretation, and to Dr Phil Studds and Mark Bell from Ramboll UK for enabling site access and sample collection. Beyond these, I am grateful to everyone that made my time at the university memorable. Ian Burke, for teaching me to remain calm in stressful situations through his unique mini bus driving style and endless supply of British Isles coffee (two not explicitly linked). Mat Evans for repeatedly allowing me to outperform him in the gym, not pressing charges in Arran, and for listening to my endless rants. Sam Allshorn, for patience and understanding despite his militaristic running of the Cohen Labs, and to Damian Howells, for always being up for a pint. I also owe huge thanks to my office mates and fellow postgraduate students for their tolerance of me over the last three years, including; Chris Hubbard, Jenn Brooke, Ross Herbert, Mike Hollaway, Cat Scott, Amber Leeson, David Tupman, Ryan Hossaini, Eimear Dunne, James Pope for not being quite as good at sport as I am, Ben Parkes for his t-shirts bringing colour into an otherwise bland office, and Bradley Jemmett-Smith and Rachael Berman for their continued friendship over the last 6 years. For having the dubious honour of sitting next to me for the last two years special thanks must go to Sarah Wallace, who willingly provided me with all office supplies, a chronological banana skin decompositional time piece, cakes, mugs, plates, cutlery and rescued me from a dire Czech situation. Thanks are also due to all those who have helped me keep in touch with some form of reality over the years; Dave McClane, Rebecca “Pearbear” Perry, Andrew “Mexican” Mason and his beautiful geometric features, Phil Rooney, Oliver Wade, Jack “Jack-bot” Ellery, Felicity Ford, Jo Minnitt, Joseph Weiler for that ticket we were just talking about, Nishal Palawan and his gatecrasher hoodie, Nick Bell 10L and his love of small animals, Simon Burton, Tom Dawson and his paddle, Matt Charles and his pear juice, Dan Faben for always wanting to get married, everyone at Bradford Bulldogs IHC, and even Eli Rosenblatt. A huge thank you to my parents for always supporting me with whatever I decided to do and for funding my various exploits over the years, and to both my brothers. Special thanks are also owed to Mr Hamshaw for spending his free time all those years ago to teach me AS-level physical geography, without which I probably would never have started university. Finally, I would also like to thank “my muse, my flame” Ashlea “gets there in the end” Henshaw for helping me get there in the end, and in the words of Jimmy MacElroy: “if you can dream it, you can do it!” This project has been jointly funded by the University of Leeds School of Earth and Environment and the John Henry Garner Endowed Scholarship. Funding for XAS analysis at the Diamond Lightsource was awarded through Burke I. T., Stewart D. I., Mortimer R. J. G. and Whittleston R. A. (2009) Micro-focus XAS study of chromium behaviour in alkaline soil-water systems. STFC Diamond Light Source AP6-1493 (4 days). ABSTRACT The poorly controlled disposal of chromium ore processing residue (COPR) is a globally widespread problem due to its potential to form chromium contaminated hyperalkaline (pH > 12) leachates. These highly oxidising leachates typically contain 2- chromium in the Cr(VI) oxidation state as its chromate anion (CrO4 ). This anion is highly mobile, toxic, carcinogenic, and exhibits a high degree of bioavailability. Under reducing conditions chromium exists in the non-toxic and poorly soluble Cr(III) oxidation state. Thus, the reduction of Cr(VI) to Cr(III) is often the goal of remediative strategies. In anaerobic subsurface environments where reducing conditions are established by the indigenous microbial population, chromium reduction can occur naturally. The microbial transformation of Cr(VI) to Cr(III) can be both a result of its direct use in microbial metabolism, or through its indirect reaction with microbially produced reduced species, e.g. Fe(II). This study has used a multidisciplinary approach to investigate the biogeochemical influences on the fate and stability of Cr(VI) leaching from a site of COPR in the north of England. Reducing sediments encountered directly beneath the COPR waste were found contain elevated concentrations of chromium. These sediments were shown to be able to remove aqueous Cr(VI) from solution when incubated with contaminated site groundwater in microcosm incubation experiments. This removal is likely a result of the abiotic reduction by soil associated microbially produced Fe(II), followed by precipitation as insoluble Cr(III) hydroxides. X-ray absorption spectroscopy (XAS) and electron microscopy confirms the association of chromium as Cr(III) with iron in these soils, hosted as a mixed Cr(III)-Fe(III) oxyhydroxide phase. Upon air oxidation, only minor amounts of chromium was remobilised from these sediments as Cr(VI). A diverse population of alkaliphilic microorganisms are indigenous to this horizon, capable of successful metabolism despite elevated pH values. This population was found to contain a consortium of microorganisms capable of iron reduction when incubated at pH 9
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