Microbial Community Characterization of Produced Water and Surrounding Seawater from Oil and Gas Production Platforms in Eastern Canada Chiu William Yeung Department of Natural Resource Sciences McGill University (Macdonald Campus) Ste-Anne-de-Bellevue, Quebec August 2010 A thesis submitted to McGill University in partial fulfillment of the requirements of the degree of Ph.D. © Chiu William Yeung, 2010 Library and Archives Bibliothèque et Canada Archives Canada Published Heritage Direction du Branch Patrimoine de l'édition 395 Wellington Street 395, rue Wellington Ottawa ON K1A 0N4 Ottawa ON K1A 0N4 Canada Canada Your file Votre référence ISBN: 978-0-494-77575-2 Our file Notre référence ISBN: 978-0-494-77575-2 NOTICE: AVIS: The author has granted a non- L'auteur a accordé une licence non exclusive exclusive license allowing Library and permettant à la Bibliothèque et Archives Archives Canada to reproduce, Canada de reproduire, publier, archiver, publish, archive, preserve, conserve, sauvegarder, conserver, transmettre au public communicate to the public by par télécommunication ou par l'Internet, prêter, telecommunication or on the Internet, distribuer et vendre des thèses partout dans le loan, distrbute and sell theses monde, à des fins commerciales ou autres, sur worldwide, for commercial or non- support microforme, papier, électronique et/ou commercial purposes, in microform, autres formats. paper, electronic and/or any other formats. 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Abstract Produced water is the largest volume of waste produced during the recovery of oil from offshore oil and gas production platforms, and is discharged directly into the surrounding marine environment. Emerging evidence suggests that offshore produced water discharges impact biota at significant distances from the point of discharge. The objectives of this study were to characterize the marine bacterial and/or archaeal populations in and around the Hibernia, Terra Nova and Thebaud production platforms, to determine if changes in the bacterial community structure might be related to the influence of the produced water, and to develop molecular methods to monitor the bacterial community structure in seawater and to track the dispersion of produced water. The marine bacterial community structure remained relatively constant (with SAB > 70%) across large distances (up to 50 km away from the platforms) and throughout several years of sampling, suggesting that the produced water discharge did not have a detectable effect on the surrounding seawater. Bacteroidetes, Alpha-proteobacteria, and Gamma-proteobacteria were the most common and abundant phyla detected in the northwestern Atlantic Ocean. The only effect potentially related to the input of produced water was found on the sediment close to the Thebaud platform, suggesting that any impact might be restricted to a small area at the sediment level adjacent to the discharge. Firmicutes was the most common and dominant phylum detected in all three produced waters, represented principally by species of Thermoanaerobacter. Both species-specific detection methods (Q-PCR and nested-PCR) detected Thermoanaerobacter in surrounding seawater within 1000 m of the production i platforms, but most of the Thermoanaerobacter were found at 100 m from Hibernia near the bottom of the water column, with only a small amount traveling into the upper water column. The properties of the produced water and existing modeling studies also support this dispersion pattern for produced water. This study provided the first characterization of the microbial diversity in and around the three major offshore oil and gas production platforms in Eastern Canada and developed new molecular methods to monitor and track the discharge of produced water. ii Résumé L‟eau de production représente le plus grand volume de déchets produits lors de la récupération du pétrole des plateformes pétrolières côtières, et cette eau est libérée directement dans le milieu marin avoisinant. Certaines évidences suggèrent que ces décharges d‟eau de production auraient un impact sur la vie marine même à une distance considérable du point de décharge. Cette étude avait pour objectifs de caractériser les populations bactériennes et/ou archéennes marines aux abords des plateformes d‟Hibernia, Terra Nova et de Thebaud, de déterminer si des changements dans la structure de la communauté procaryotique peuvent être reliés à l‟influence de l‟eau de production, de développer des outils moléculaires pour le suivi des communautés procaryotique dans l‟eau de mer, et finalement, de retracer la dispersion de l‟eau de production. La structure de la communauté bactérienne marine est demeurée relativement constante (SAB > 70%) sur de longues distances (jusqu‟à 50 km des plateformes) et sur plusieurs années d‟échantillonnage. L‟eau de production n‟aurait donc pas d‟effet détectable sur la structure de la communauté bactérienne environnante. Les Bacteroidetes, Alpha-proteobacteria et Gamma-proteobacteria représentaient les phyla les plus communs et abondants dans le Nord-Ouest de l‟océan Atlantique. Le seul effet possiblement relié à l‟introduction de l‟eau de production fut observé dans les sédiments près de la plateforme Thebaud, ce qui suggère que l‟impact pourrait se limiter à l‟étroite région au niveau des sédiments adjacents à la décharge. Les Firmicutes constituaient le phylum dominant détecté dans les trois eaux de production et étaient représentés principalement par des espèces de iii Thermoanaerobacter. Deux méthodes moléculaires spécifiques à l‟espèce (Q- PCR et PCR-niché) ont pu détecter Thermoanaerobacter dans l‟eau de mer dans un rayon de 1000 m des plateformes de production, mais la majorité des Thermoanaerobacter trouvés à 100 m d‟Hibernia se situait au bas de la colonne d‟eau, avec seulement une petite quantité migrant plus haut dans la colonne. Les propriétés de l‟eau de production et les modèles existants supportent également le profil de dispersion pour l‟eau de production. Cette étude rapporte la première caractérisation de la diversité microbienne aux abords de trois plateformes côtières de production de pétrole et gaz de l‟est du Canada et le développement de nouveaux outils moléculaires pour retracer et suivre la libération de l‟eau de production. iv ACKNOWLEDGEMENTS First I would like to thank Dr. Charles W. Greer for allowing me to study in his lab and for all his support in the projects. I would also like to thank Dr. Ken Lee for introducing me to the project, for all the support in the project and the freedom to do the work. Dr. Whyte, Dr. Driscoll, Dr. Niven, Dr. Chénier and other members of my committees greatly improved the quality of my work by constantly pushing me to perform better. My NRC lab-mates Dr. Nancy Perrault, Dr. Louis Jugnia, Dr. Marc Auffret, Terry Bell, Nathalie Fortin, Claude Masson, and Sylvie Sanschagrin were all very helpful in my Ph.D. studies. I would like to thank my COOGER lab-mates Dr. Paul Kepkay, Dr. Zhengkai Li, Dr. Haibo Niu, Jay Bugden, Lorraine Hamilton, Tom King, Susan Cobanli, Brent Law, Byron Amirault, and Brian Robinson for all their support in Halifax. I would also like to acknowledge the organizations, NRCan PERD, DFO and NRC, for their financial support. Finally, none of this would have been possible without the support and encouragement of my parents and my wife. It is to them that I dedicate this thesis. Thank you! v CONTRIBUTIONS TO KNOWLEDGE This study presents the first culture-independent analysis of the bacterial and archaeal communities present in produced water originating from three eastern Canada oil and gas production platforms (Hibernia, Terra Nova and Thebaud). This study presents the first culture-independent analysis of the bacterial communities present in the surrounding seawater adjacent to three eastern Canada oil and gas production platforms (Hibernia, Terra Nova and Thebaud). This is the first marine bacterial and/or archaeal community analysis study of three ecologically and commercially important areas: the Grand Banks, Sable Island Bank and the Gully (the first Marine Protected Area in the Eastern Canada) in the western Atlantic Ocean. The first marine 16S rRNA gene-based taxonomic microarray was developed for rapid detection, identification and profiling of the bacterial diversity in seawater and in produced waters. Sensitive Q-PCR and nested-PCR methods were developed to track specific target bacteria associated with produced waters from Hibernia and Terra Nova in the marine environment. This study is the first to quantitatively monitor a natural component of the produced water, the bacterium Thermoanaerobacter spp., in the surrounding seawater, allowing