PERFORMANCE ASSESSMENT AND ENRICHMENT OF ANAEROBIC METHANE OXIDISING MICROBIAL COMMUNITIES FROM MARINE SEDIMENTS IN BIOREACTORS Susma Bhattarai Gautam Joint PhD degree in Environmental Technology UNIVERSITE--------------- — PARIS-EST Docteur de I’Universite Paris-Est Speciality : Science et Technique de I’Environnement Dottore di Ricerca in Tecnologie Ambientali UNESCO-IHE Institute for Water Education Degree of Doctor in Environmental Technology These - Tesi di Dottorato - PhD thesis Susma Bhattarai Gautam Performance Assessment and Enrichment of Anaerobic Methane Oxidizing Microbial Communities from Marine Sediments in Bioreactors Defended on December 16th, 2016 In front of the PhD committee Dr. Cartsen Vogt Reviewer Dr. Artin Hatzikioseyian Reviewer Prof. dr. ir. Piet. N. L. Lens Promotor Prof. Michel Madon Co-Promotor Prof. Giovanni Esposito Co-Promotor Dr. Eldon R. Rene Co-Promotor Hab. Dr. Eric van Hullebusch Examiner 3 ETeC oS Erasmus Joint doctorate programme in Environmental Technology for Contaminated Solids, Soils and Sediments (ETeCoS3] Thesis Committee Thesis Promoter Prof. dr. ir. Piet N. L. Lens Professor of Environmental Biotechnology UNESCO-IHE, Institute for Water Education Delft, the Netherlands Thesis Co-Promoters Prof. Michel Madon University of Paris-Est, Institut Francilien des Sciences Appliqueees Champs sur Marne, France Dr. Giovanni Esposito Associate Professor of Sanitary and Environmental Engineering University of Cassino and Southern Lazio, Cassino, Italy Dr. Eldon R. Rene UNESCO-IHE, Institute for Water Education Delft, the Netherlands Examiner Hab. Dr. Eric van Hullebusch Associate Professor of Biogeochemistry University of Paris-Est, Institut Francilien des Sciences Appliqueees Champs sur Marne, France Reviewers Dr. Cartsen Vogt Helmholtz Centre for Environmental Research - UFZ Department of Isotope Biogeochemistry Leipzig, Germany Dr. Artin Hatzikioseyian National Technical University of Athens Athens, Greece This research was conducted under the auspices of the Erasmus Mundus Joint Doctorate Environmental Technologies for Contaminated Solids, Soils, and Sediments (ETeCoS3) and The Netherlands Research School for the Socio- Economic and Natural Sciences of the Environment (SENSE). CRC Press/Balkema is an imprint of the Taylor & Francis Group, an informa business © 2018, Susma Bhattarai Gautam All rights reserved. No part of this publication or the information contained herein may be reproduced, stored in a retrieval system, or transmitted in any form or by any means, electronic, mechanical, by photocopying, recording or otherwise, without written prior permission from the publishers. Although all care is taken to ensure the integrity and quality of this publication and information herein, no responsibility is assumed by the publishers or the author for any damage to property or persons as a result of the operation or use of this publication and or the information contained herein. Published by: CRC Press/Balkema PO Box 11320, 2301 EH Leiden, The Netherlands e-mail: [email protected] www.crcpress.com - www.taylorandfrancis.com ISBN: 978-1 -138-33021 -4 (Taylor & Francis Group) Table of contents Chapter Title Page no. Acknowledgements xi Summary xiii Samenvatting xv Somario xvii Resume xix CHAPTER 1 General Introduction and Thesis Outline..................................................1 1.1 General introduction and problem statement.....................................................2 1.2 Objectives and scope of the study.....................................................................4 1.3 Outlines of this thesis.........................................................................................5 1.4 References..........................................................................................................6 CHAPTER 2 Physiology and Distribution of Anaerobic Oxidation of Methane by Archaeal Methanotrophs................................................................................................11 2.1 Introduction........................................................................................................12 2.2 Microbiology of anaerobic methane oxidation.................................................15 2.3 Physiology of AN ME..........................................................................................23 2.4 Drivers for distribution of ANME in natural habitats........................................29 2.5 Ex situ enrichment of ANME.............................................................................40 2.6 Approaches for AOM and ANME studies........................................................49 2.7 Conclusions and outlook..................................................................................57 2.8 References........................................................................................................58 CHAPTER 3 Microbial Sulfate Reducing Activities in Anoxic Sediment from Marine Lake Grevelingen..........................................................................................................83 Abstract.........................................................................................................................84 3.1 Introduction........................................................................................................84 3.2 Materials and Methods.....................................................................................86 3.3 Results..............................................................................................................90 3.4 Discussion.........................................................................................................97 3.5 Conclusions.....................................................................................................101 3.6 References......................................................................................................101 CHAPTER 4 Anaerobic Methane Oxidizing Microbial Community in a Coastal Marine Sediment: Anaerobic Methanotrophy Dominated by ANME-3..................................107 4.1 Introduction......................................................................................................108 vii 4.2 Materials and methods....................................................................................111 4.3 Results.............................................................................................................116 4.4 Discussion.......................................................................................................127 4.5 Conclusions.....................................................................................................134 4.6 References......................................................................................................134 CHAPTER 5 Enrichment of Anaerobic Methane Oxidizing ANME-1 from Ginsburg Mud Volcano (Gulf of Cadiz) Sediment in a Biotrickling Filte.....................................145 5.1 Introduction......................................................................................................146 5.2 Materials and methods....................................................................................148 5.3 Results.............................................................................................................155 5.4 Discussion.......................................................................................................163 5.5 Conclusions.....................................................................................................167 5.6 References.....................................................................................................168 CHAPTER 6 Enrichment of ANME-2 Dominated Anaerobic Oxidation of Methane Coupled to Sulfate Reduction Consortia from Cold Seep Sediment (Ginsburg Mud Volcano, Gulf of Cadiz) in a Membrane Bioreactor....................................................177 6.1 Introduction......................................................................................................178 6.2 Materials and Methods....................................................................................181 6.3 Results.............................................................................................................188 6.4 Discussion.......................................................................................................194 6.5 Conclusions.....................................................................................................198 6.6 References......................................................................................................198 CHAPTER 7 Response of Highly Enriched ANME-2a Community to the Different Pressure and Temperature Conditions......................................................................207 7.1 Introduction......................................................................................................208 7.2 Materials and methods...................................................................................210 7.3 Results............................................................................................................213 7.4 Discussion.......................................................................................................220 7.5 Conclusions.....................................................................................................223 7.6 References......................................................................................................224 CHAPTER 8 General Discussion...............................................................................231 8.1 Introduction......................................................................................................232 8.2 Anaerobic oxidation of methane in coastal sediment.....................................232 8.3 Anaerobic oxidation of methane in bioreactors...............................................236 8.4 Enrichment of ANME in different bioreactor configurations..........................238 8.5 Future perspectives in ANME enrichments...................................................240 8.6 References......................................................................................................241 Biography....................................................................................................................247 Publications.................................................................................................................248 SENSE diploma for specialized PhD trainings..........................................................249 ix