https://theses.gla.ac.uk/ Theses Digitisation: https://www.gla.ac.uk/myglasgow/research/enlighten/theses/digitisation/ This is a digitised version of the original print thesis. Copyright and moral rights for this work are retained by the author A copy can be downloaded for personal non-commercial research or study, without prior permission or charge This work cannot be reproduced or quoted extensively from without first obtaining permission in writing from the author The content must not be changed in any way or sold commercially in any format or medium without the formal permission of the author When referring to this work, full bibliographic details including the author, title, awarding institution and date of the thesis must be given Enlighten: Theses https://theses.gla.ac.uk/ [email protected] EVALUATION OF METALLOTHIONEIN AS AN ECOTOXICOLOGICAL BIOMARKER IN NUCELLA LAPILLUS AND LITTORINA LITTOREA Kenneth M eI-Yee Leung Presented in candidature for the degree of Doctor of Philosophy, to the Institute of Biomedical and Life Sciences, University of Glasgow. March, 2000 ProQuest Number: 10390844 All rights reserved INFORMATION TO ALL USERS The quality of this reproduction is dependent upon the quality of the copy submitted. In the unlikely event that the author did not send a complete manuscript and there are missing pages, these will be noted. Also, if material had to be removed, a note will indicate the deletion. uest ProQuest 10390844 Published by ProQuest LLO (2017). C o pyright of the Dissertation is held by the Author. All rights reserved. This work is protected against unauthorized copying under Title 17, United States Code Microform Edition © ProQuest LLO. ProQuest LLO. 789 East Eisenhower Parkway P.Q. Box 1346 Ann Arbor, Ml 4 8 1 0 6 - 1346 GLASGOW 1 , UNIVERSITY ' 'L IB R A R Y , In Memory of JAMES HENRY SCOTT UNIVERSITY John Swire & Sons Ltd. GLASGOW Hospital for Marine Invertebrates 0 Dr. Ecotoxicology don't know what I may seem to the world, but, as to myself, I seem to have been only like a boy playing on the sea shore, and diverting myself in now and then finding a smoother pebble or a prettier shell than ordinary, whilst the great ocean of truth lay all undiscovered before me.” Sir Isaac Newton (1642-1727) Source; Carey J. (1995) The Faber Book of Science, f. f., London, pp. 30-34. P- dfWUiUt TWO-DOMAIN M l STRUCTURE (Romero et al. 1996, J Molecular Modelling 11:417-426) Candidate's Declaration I declare that the work recorded in this thesis is entirely my own, unless otherwise stated and that it is of my own composition. No part of this work has been submitted for any other degree. Kenneth Mei-Yee Leung March 2000 ^ i Dedication This thesis is dedicated with love and respect to my parents, Lam and Kam-Chu Leung ii ^ Acknowledgements I am greatly indebted to John Swire and Sons Ltd. for providing me with a James Henry Scott Scholarship that made this study possible. I would like to thank Sir Edward Scott (Director of John Swire and Sons Ltd.), Mr Graham Docherty and Ms Winnie Wong for their caring, friendship, wonderful arrangement and hospitality. I would like to acknowledge Prof. Mike Cowling for finding me the most suitable supervisor and arranging my registration in the earhest stage. Thanks and gratitude are due to my supervisor. Prof. Bob Furness for his continuous, generous help and advice, which made em invaluable contribution to this study. (Bob showed me all the possible sampling sites and helped sampling for the preliminary study, read through all my work even when he was very ill, and found funding for me to attend the SET AC conference and enjoy bird-watching in Germany during summer 1999). Thanks are also due to Dr Alan Taylor for allowing me to work and use all the facilities in his physiology laboratory, and for giving up his valuable time to me. He also taught me how to measure haemocyanin and oxygen carrying capacity of haemolymph in the blood samples of dogwhelks; and co-authored Chapter 3. I am also grateful to June Freel for providing efficient and excellent technical support throughout this study, helping me to set and sample Chelex and mussels, and teaching me about instrumentation and biochemical assays as well as Scottish language and culture (her cheerful smiles made the routine biochemical assay "a wee bit" more enjoyable), I would also like to thank die aquarium manager, John Laurie and store manager William Orr for their kindness and wonderful assistance over the last 3 years. Thanks to all the people who assisted with this study and taught me in different aspects. To Drs Ben Ross, Stuart Bearhop, Ian Morgan, Brendan Codley and Ms Andrea Fidgett, who improved the English of my written work, provided useful criticism and marvellous encouragement. To Drs Ian Morgan and Paul Parslow- Williams for showing me how to quantify proteins and RNA of muscle samples. To Dr Michael Burrows for the enthusiastic discussion and all tlie important information about dogwhelks. To Drs Neil Metcalfe and Ruedi Nager for making themselves ^ iii available and being patient when answering my statistical queries. To Drs Laurence Tetley and Hugh Elder, for allowing me to join their undergraduate course on "Principle and application of electi'onic microscopy" and letting me use the SEM. To Ms Margaret Mullm and Kate Orr for teaching and helping me to prepare tissue samples for histology. To Dr Jim Atkinson, for allowing and assisting me to set the Chelex and mussels at the pier of University Marine Biological Station Millport. To Dr Douglas Neil and Crant Stentiford, for all the enjoyable and fruitful discussions, and allowing me to be involved in the Nephrops project. Crateful thanks are also due to aU of my field assistants including Scott Ramsey, Mark Cottam, K.Y. Tong, Y.T. Chan, Diane Lu, Andrew Ng, and Louise Hung. For me it was a pleasure to study for a PhD in Glasgow, but without the support of many, colleagues and friends it would have been a much more difficult task. In particular I must thank my fellows: David Stevens, Stuart Bearhop, J.P. Granadeiro, Brendan Codley, Bernard Zonfrillo, Stuart Fleming, Steve Arnott, Crant Stentiford, Steve Votier, Jon Blount; aU my office mates: Ben Ross (my principal English teacher), Andrea Fidgett, Scott Ramsey, Mark Cottam, Annette Broderick, Isabel Coombs, Harriet Torlesse, Helen Whitaker, Mike Fowler, Henriikka Clarkeburn; all the other postgraduates and members of staff in the department. Thanks to all my friends in Glasgow, especially Ho-Man Chan, Raymond K.Y. Tong, aU brothers and sisters of CCCC. I thank all these people again for the laughter, sharing and precious friendships — without them, my PhD life would not be so fruitful, enjoyable and unforgettable. Many of my teachers and friends overseas, particularly in Hong Kong, helped me at one stage or another during the course of this study. Thanks to aU wonderful colleagues in the BCH Department of CityUHK. To Prof Wu, for his continuous encouragement, support and advice in many ways, and funding me to attend the international conference on marine pollution and ecotoxicology at CityUHK during summer 1998. To Dr Paul Lam, for showing me the advertisement about the James Hemy Scott Scholarships in tiie first place, and telling me about the excellent postgraduate course on "Conceptual and Analytical Approached to Problem Solving in Ecotoxicology" at Roskilde University of Denmark (March 1997). I must tliank Dr Valery Forbes (Roskilde University), for allowing me to take this course without paying the tuition fee. What I learned from this course was extremely useful for my PhD. Thanks are also due to Dr S.C. Cheung, for giving me invaluable advice on how iv to tag gastropods. I am indebted to King-Tai Wo, Drs Paul Lam and Graham Blackmore for their continuous help, particularly by always sending me any information that I needed. I also acknowledge Prof Wu and Dr T.C. Lau who collaborated for the Chelex project and performed metal analyses on the Chelex and mussels samples (most of the data are still being processed and therefore not included in this tiiesis). Although I could not include the research results obtained from Iceland in this thesis (most samples await metal and biochemical analyses), this trip to Iceland was one of the most important milestones in my PhD study. I am especially grateful for the TMR research grant (as a part of BIOICE project from EC) that allowed me to stay and work at the Sandgerdi Marine Centre (SMC), and carry out laboratory and field studies during August to November 1999. I am grateful for die enthusiastic support provided to me by Mr Gudmundur and Prof Jorundur Svavarsson. In particular. Prof Svavarsson patiently taught me how to identify of the various stages of imposex in dogwhelks, generously offered financial support for my field trips in Iceland (sampling of dogwhelks from South to the very North - Grimsey Island) and collaborated for the project. Thanks are also due to all colleagues and lovely people at Sandgerdi, especially Halldor, Reynir, Oli, Stebbi, Binna, Ragnar, Stuart and Ann, for their great help and wonderful friendship. I would like to thank my family, especially my sister (L.M. Leung) and her family (S.H. Wong, Y.Y. Wong and H.Y. Wong) as well as my best friends (C.W. Yu, C.L. Lam, C.L. Hui, W.L. Chan, and their families) for their loving care, friendship, strong and constant support in many ways. All these amazing graces have already been engraved in my thankful mind. Certainly, last but not least, I must especially thank my dearest Diane, as always, my source of strength and the colours of my life.