Effects of Environmental Stressors on the Black Bream
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Effects of environmental stressors on the black bream (Acanthopagrus butcheri). A thesis submitted in fulfilment of the requirements for the degree of Doctor of Philosophy Kathryn L. Hassell B. App. Sci (Hons) School of Applied Sciences College of Science, Engineering and Technology RMIT University September 2009 DECLARATION The contents of this thesis are a result of work undertaken whilst officially enrolled for the degree of Doctor of Philosophy in the School of Applied Sciences, Department of Biotechnology and Environmental Biology of the College of Science, Engineering and Technology at the Royal Melbourne Institute of Technology. All procedures involving vertebrate animals (fish) were conducted in accordance with the RMIT University Animal Ethics Committee, under the approved project AEC 0523. To the best of my knowledge, all work performed by others, published or unpublished, has been duly acknowledged in this thesis. This thesis has not been submitted, in whole or in part, to qualify for any other award. Signed:………………………………… Date:………………………………… ii ACKNOWLEDGEMENTS I would like to thank my three supervisors for all their support and encouragement throughout the duration of this PhD project. To Professor Dayanthi Nugegoda, thankyou for initially encouraging me and providing the opportunity to take on this project and for your support throughout. To Dr. Patrick Coutin, thankyou for your support, encouragement and enthusiasm, especially during the really challenging times. Your rapid responses to questions and queries, and the fact that you always made yourself available when I wanted to discuss my progress is particularly appreciated. To Sue Codi King, thankyou for all of your assistance with laboratory techniques and for all the long phone calls to discuss results and provide advice, support and encouragement. Thankyou to Dianne Rose and Dr. Warren Davies of the Victorian Environmental Protection Authority (EPA) for providing financial support to undertake this PhD project, as well as Dr. Vincent Pettigrove of Melbourne Water for assisting with field collections and providing the resources to do the field work. I would like to acknowledge Sven Inge Kristiansen and Janne Eidem of Biosense Laboratories (HIB-Thormøhlensgt 55, N-5008 Bergen, Norway) for conducting the initial screening of black bream plasma samples to establish a suitable vitellogenin primary antibody. I would like to thank all the fishermen that assisted with field collections throughout the project, since without their help I couldn’t have achieved the work. Thanks to Doc McKenzie, Dirk Erler, Geordie King and Frank Moletta for assistance in the Gippsland Lakes, to John Wood, John McGuckin and Ron Lewis for assistance in Mallacoota Inlet, to Anthony Hocking for assistance in the Yarra River and Maribyrnong River and Phil McAdam for assistance in the Werribee River and Little River. Thankyou to the staff at the Victorian Marine Science Consortium and DPI Queenscliff Centre who were so helpful to me whilst working in the Queenscliff laboratories. A iii special thanks to Liz McGrath for helping with all the administration details, to Rod Watson for helping me set up the laboratory to conduct the experiments, to Lachlan McKinnon and Nathan O’Mahony for helping with the management of broodstock in the Aquaculture facility, Mayumi and Graeme Allinson for measuring estrogens in water samples and finally to Kyne Krusic-Golub and Therese Stokie for analysing fish otoliths and providing age estimates for all the field collected black bream. Thankyou also to Shannon Fernandes at RMIT University for technical assistance throughout the project. Thankyou also to all the other postgraduate students that have been there to share in both the good and the bad sides of the PhD experience. Knowing that others are going through the same suite of ups and downs was both relieving and encouraging. Thankyou to the group at VMSC, Lorenz Frick, Tim Smith, Fabian Trinnie, David Phillips, Isla Fitridge, Fiona Warry, Peter Macreadie, Joel Williams, Matt Reardon, Justin Bell, Daniel Grixti and Corey Green, you have all contributed to the positive experience of working in the VMSC labs. Thanks also to everyone from the RMIT Ecotoxicology lab, especially Andrew Harford, Wilson Lennard, Lisa Toogood, Liz Fields and Scott Raymond for friendship and advice in the early stages of my project, to other students in the Ecotoxicology lab both past and present, for friendship, support and encouragement, as well as Fiona Baird and Edward Grixti for providing advice on immunological techniques and procedures. Finally I would like to say a big thankyou to my family for providing such great love and support throughout. Mum and Dad, Lloyd and Caroline you have all been so encouraging and supportive of my work despite it often meaning that I have had to spend less time with you. And to Nathan, I am most grateful of your love and support. You have been with me throughout the entire experience, and your encouragement and understanding has helped me stay grounded during the tough times and really contributed greatly to me completing this thesis and achieving my goals. iv PUBLICATIONS/CONFERENCES PUBLICATIONS • Hassell, K. L., Coutin, P. C. and Nugegoda, D. (2009) “A novel approach to controlling dissolved oxygen levels in laboratory experiments.” Journal of Experimental Marine Biology and Ecology, 371, 147-154. • Hassell, K. L., Toogood, L. G., Coutin, P. C. and Nugegoda, D. “Embryo development in the Australian black bream Acanthopagrus butcheri (Munro).” Aquaculture (submitted September, 2009). • Hassell, K.L., Coutin, P.C. and Nugegoda, D. (2008). Hypoxia, low salinity and lowered temperature reduce embryo survival and hatch rates in black bream (Acanthopagrus butcheri, Munro 1949). Journal of Fish Biology, 72, 1623-1636. • Hassell, K.L., Coutin, P.C. and Nugegoda, D. (2008). Hypoxia impairs embryo development and survival in black bream (Acanthopagrus butcheri). Marine Pollution Bulletin, 57, 302-306. • Codi King, S, Hassell, K.L., Nugegoda D. and Kristiansen, S. I. (2008). The assessment of vitellogenin as a biomarker of exposure to estrogenic compounds in two Australian Perciformes. Marine Environmental Research, 66, 116-118. v CONFERENCES • Kathryn Hassell, Patrick Coutin, Dayanthi Nugegoda. “The effects of environmental stressors on aspects of reproduction in black bream, Acanthopagrus butcheri”. Australian Society for Fish Biology (ASFB) Conference, Sydney, Australia. September 2008. (ORAL PRESENTATION) • Kathryn Hassell, Vincent Pettigrove and Dayanthi Nugegoda. “Black bream (Acanthopagrus butcheri) as a potential biomonitoring species for endocrine disrupting chemicals in estuaries of southern Australia”. Society for Ecotoxicology and Chemistry (SETAC) 5th World Congress, Sydney, Australia. August, 2008. (ORAL PRESENTATION) • Kathryn Hassell, Patrick Coutin, Dayanthi Nugegoda. “Black bream embryo survival and development is impaired by exposure to hypoxia”. 5th International Conference on Marine Pollution and Ecotoxicology (ICMPE), Hong Kong, Hong Kong. June, 2007. (ORAL PRESENTATION) • Susan Codi King, Kathryn Hassell, Dayanthi Nugegoda. “Induction of vitellogenin in barramundi and black bream as a biomarker of exposure to EDCs in Australian waters”. 5th International Conference on Marine Pollution and Ecotoxicology (ICMPE), Hong Kong, Hong Kong. June, 2007. (ORAL PRESENTATION) vi • Kathryn Hassell, Patrick Coutin, Dayanthi Nugegoda. “Climate change: impacts on embryos of the estuarine black bream”. Australian Marine Sciences Association (AMSA) Conference, Melbourne, Australia. June, 2007. (ORAL PRESENTATION) • Kathryn Hassell, Patrick Coutin, Dayanthi Nugegoda. “Mild hypoxia, lowered temperature and decreased salinity levels affect embryo survival and hatching in black bream, Acanthopagrus butcheri”. Australian Society of Fish Biology (ASFB) Conference, Tasmania, Australia. September, 2006. (ORAL PRESENTATION) vii TABLE OF CONTENTS Declaration.............................................................................................................................................ii Acknowledgements...............................................................................................................................iii Publications/Conferences......................................................................................................................v Table of Contents...............................................................................................................................viii List of Tables.......................................................................................................................................xv List of Figures....................................................................................................................................xvi List of Abbreviations..........................................................................................................................xx Summary................................................................................................................................................1 CHAPTER 1: INTRODUCTION................................................................................................................. 6 1.1 Environmental stressors……………………………………………….…………................. 7 1.1.1 Dissolved oxygen and hypoxia……………………………………...………………............... 7 1.1.1.1 Physiological effects of hypoxia on the early life stages of fishes………….………............... 11