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Escola d’Enginyeria
Departament d'Enginyeria Química, Biològica i Ambiental
“Anaerobic microbial transformation of chlorinated alkanes in cultures derived from Besòs River estuary sediments”
PhD Thesis
Siti Hatijah Mortan
Doctorat en Ciència i Tecnologia Ambientals
Supervised by
Dr Ernest Marco Urrea
Dra Gloria Caminal Saperas
Bellaterra, Cerdanyola del Vallès, Barcelona
March 2017
Title: Anaerobic microbial transformation of chlorinated alkanes in cultures derived from Besòs River estuary sediments
Presented by: Siti Hatijah Mortan
Supervised by: Ernest Marco Urrea and Gloria Caminal Saperas
PhD program in Environmental Science and Technology Departament d'Enginyeria Química, Biològica i Ambiental Escola d’Enginyeria Universitat Autònoma de Barcelona, Bellatera, 2017
This work was supported by the Xarxa de Referència en Biotecnologia de la Generalitat de Catalunya (Premi Pot d’Idees 2016), the Spanish Ministry of Economy and Competitiveness and FEDER(project CTM2013-48545-C2-1-R) and supported by the Generalitat de Catalunya (Consolidated Research Group 2014-SGR-476). The author acknowledges the predoctoral fellowship from Ministry of Higher Education Malaysia and Universiti Malaysia Pahang.
Part of this work has been done in collaboration with the Department Isotope Biogeochemistry, Helmholtz Centre for Environmental Research-UFZ, Germany (Dr. L. Adrian); Departament de Genètica i Microbiologia, Universitat Autònoma de Barcelona (Dra. N. Gaju, Dra. M. Martínez, E. Parladé); Departament de Cristal-lografia, Mineralogia i Dipòsits Minerals, Facultat de Geologia, Universitat de Barcelona (Dra. M. Rosell, Dr. J. Palau); Department of Earth and Environmental Sciences, University of Waterloo, Canada (Dr. O. Shouakar-Stash).
ERNEST MARCO URREA, Professor Agregat Interí del Departament d'Enginyeria Química,
Biològica i Ambiental de la Universitat Autònoma de Barcelona, GLORIA CAMINAL
SAPERAS, Investigador Científic del Institut de Química Avançada de Catalunya (IQAC) del
CSIC,
CERTIFIQUEM:
Que el titulat en Enginyeria Bioquímica Siti Hatijah Mortan ha realitzat sota la nostra direcció, en els laboratoris del Departament d'Enginyeria Química, Biològica i Ambiental el treball que amb el títol “Anaerobic microbial transformation of chlorinated alkanes in cultures derived from Besòs River estuary sediments”, es presenta en aquesta memòria, la qual constitueix la seva Tesi per optar al Grau de Doctor per la Universitat Autònoma de Barcelona.
I perquè en prengueu coneixement i consti als efectes oportuns, presentem a l’Escola d’Enginyeria de la Universitat Autònoma de Barcelona l’esmentada Tesi, signant el present certificat a Bellaterra, Marc 2017.
Dr. Ernest Marco Urrea Dra. Gloria Caminal Saperas
ACKNOWLEDGEMENT
First and foremost, a very special gratitude to Dr Ernest Marco and Dr Gloria Caminal Saperas for being such a wonderful and helpful supervisors to me. Thank you very much for your guidance and I am highly indebted to both of you for all the encouragement, the patience for my shortcomings and open up the opportunity for me to work in such an interesting research.
A special thank you to Dr Lucia Martin-Gonzalez for took a great care of me in the lab and Alba Trueba-Santiso whom I’ve shared so many fun and memorable times working with the ‘anaerobic babies’. I would like to take this moment also to express my appreciation to Dr Teresa Vicent and everyone in “Toxics” Research Group, UAB for your continuous support and I am very grateful being a member of this extremely supportive research group.
Special mention to Dr Lorenz Adrian and his research group in the Department Isotope Biogeochemistry (UFZ), Leipzig, Germany for the opportunity to do my research stay in your lab. Working in UFZ was a great joy and experience, thanks to this wonderful group of people. Thank you very much also to Dr Monica Rosell from Departament de Mineralogia (UB), Dr Ivonne Nijenhuis (UFZ), Dr Nuria Gaju, Dr Maira Martinez-Alonso and Eloi Parlade from Departament de Genètica i Microbiologia (UAB) for our research collaboration.
I also want to thanks the members of the Departament d'Enginyeria Química, Biològica i Ambiental and Institut de Ciència i Tecnologia Ambiental (ICTA) for their helpful assistance. I would like to acknowledge Manuel Plaza and Lucia Delgado for their technical assistance.
Sincere thanks are extended to the examination committee for reviewing this thesis.
Last but not least, special thanks to my family and good friends who have provided me their support and understanding throughout this whole PhD journey.
‘Gracias a todos! Gracies a tothom!’
Abstract
ABSTRACT
Halogenated compounds (organohalides) are recalcitrant organic compounds, exhibiting
toxic effects on human health and the ecosystem. Among the organohalides, chlorinated alkanes
such as chloroethanes and chloropropanes received less attention despite their large use in the
industry and have caused serious environmental problems. Physicochemical processes are the
most commonly used technology to treat groundwaters contaminated with these compounds,
however these techniques imply high operational costs and require a post-treatment step to
completely destroy these compounds. Biodegradation has become a more suitable, cost-efficient
and environmental friendly technology to address contamination of these compounds.
In this study, we aimed to obtain an enrichment culture containing organohalide-respiring
bacteria able to transform halogenated alkanes. A stable nonmethanogenic enriched anaerobic
culture that exclusively dehalogenates vicinally chlorinated and brominated alkanes via dihaloelimination was established from Besòs River estuary sediments (Barcelona, Spain).
Application of genus specific primers targeting 16S rRNA gene sequences together with the observation of physiological characteristics of the dechlorinating culture indicated that a
Dehalogenimonas strain was the responsible for chlorinated alkane degradation in the
microcosms. The increase of Dehalogenimonas 16S rRNA gene copies using quantitative PCR
(qPCR) revealed that 1,2-dichloropropane (1,2-DCP) dechlorination was coupled to
Dehalogenimonas growth in this culture.
Carbon and dual carbon-chlorine (C-Cl) isotope fractionation during anaerobic
biodegradation of 1,2-DCP and 1,2-dichloroethane (1,2-DCA), respectively, by
Dehalogenimonas-containing enrichment culture were determined in this study. Compound
specific isotope analysis revealed that the Dehalogenimonas-catalyzed carbon isotopic
i
Abstract