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Claire Michelle Volschenk COPYRIGHT AND CITATION CONSIDERATIONS FOR THIS THESIS/ DISSERTATION o Attribution — You must give appropriate credit, provide a link to the license, and indicate if changes were made. You may do so in any reasonable manner, but not in any way that suggests the licensor endorses you or your use. o NonCommercial — You may not use the material for commercial purposes. o ShareAlike — If you remix, transform, or build upon the material, you must distribute your contributions under the same license as the original. How to cite this thesis Surname, Initial(s). (2012). Title of the thesis or dissertation (Doctoral Thesis / Master’s Dissertation). Johannesburg: University of Johannesburg. Available from: http://hdl.handle.net/102000/0002 (Accessed: 22 August 2017). TOXICITY TESTING FOR THE ESTABLISHMENT OF LC50 AND EC50 VALUES FOR TECHNICAL GRADE DDT, IN CONJUNCTION WITH BIOMARKERS AND BIO-ACCUMULATION IN SYNODONTIS ZAMBEZENSIS. BY CLAIRE MICHELLE VOLSCHENK THESIS SUBMITTED IN FULFILMENT OF THE REQUIREMENTS OF THE DEGREE PHILOSOPHIAE DOCTOR IN ZOOLOGY IN THE FACULTY OF SCIENCE AT THE UNIVERSITY OF JOHANNESBURG SUPERVISOR: PROF. RICHARD GREENFIELD CO-SUPERVISOR: PROF. JOHANNES H.J VAN VUREN NOVEMBER 2018 TABLE OF CONTENTS LIST OF FIGURES .................................................................................................... vii LIST OF TABLES ....................................................................................................... ix LIST OF ABBREVIATIONS ........................................................................................ x ACKNOWLEDGEMENTS ........................................................................................ xvi SUMMARY ............................................................................................................. xviii Chapter 1: General introduction ............................................................................. 1 1. General introduction ............................................................................................ 1 1.1 Background.................................................................................................... 2 1.2 Biological markers and selected indicator species ......................................... 3 1.3 Motivation for study ........................................................................................ 4 1.4 Hypotheses, aims and objectives .................................................................. 5 1.4.1 Hypotheses ............................................................................................. 6 1.4.2 Aims ........................................................................................................ 6 1.4.3 Objectives ................................................................................................ 7 1.5 Layout of thesis .............................................................................................. 7 Chapter 1: General introduction ....................................................................... 7 Chapter 2: Literature review ............................................................................. 8 Chapter 3: Focus species ................................................................................. 8 Chapter 4: Artificial breeding and embryonic development of Synodontis zambezensis (Peters, 1852) ............................................................................. 8 Chapter 5: Baseline bio-accumulation concentrations and resulting oxidative stress in Synodontis zambezensis after an acute laboratory exposure to 4, 4’- DDT .................................................................................................................. 9 Chapter 6: Biomarkers of exposure and energetics ......................................... 9 Chapter 7: Ribonucleic acid (RNA) as a biomarker .......................................... 9 Chapter 8: LC50 exposures and species sensitivity distributions ...................... 9 Chapter 9: Conclusions and recommendations ................................................ 9 Chapter 10: References ................................................................................... 9 Appendix A: Raw data ...................................................................................... 9 Appendix B: Published papers ....................................................................... 10 Appendix C: Conference contributions ........................................................... 10 1.6 Anticipated contribution ............................................................................... 10 i | P a g e Chapter 2: Literature review .................................................................................. 11 2. Introduction ....................................................................................................... 11 2.1 4,4’-DDT ...................................................................................................... 12 2.1.1 Historical use of 4,4’-DDT ...................................................................... 12 2.1.2 The use of 4,4’-DDT in South Africa ...................................................... 14 2.1.3 Intended target of 4,4’-DDT ................................................................... 15 2.1.4 The fate of 4,4’-DDT in the environment ................................................ 16 2.2 Bio-accumulation ......................................................................................... 18 2.2.1 Absorption and regulation ...................................................................... 19 2.3 Effects of 4,4’-DDT on fish and offspring ..................................................... 20 2.3.1 Developmental toxicity ........................................................................... 21 2.4 Toxicity testing framework ........................................................................... 22 2.4.1 Acute exposure...................................................................................... 22 2.4.2 Acute fish toxicity test (OECD TG, 203) ................................................ 23 2.5 Biomarkers................................................................................................... 23 2.5.1 Biomarkers of exposure ......................................................................... 24 Cytochrome P450 (CYP450) ....................................................................... 24 Acetylcholinesterase (AChE)....................................................................... 25 Cellular energy allocation (CEA) ................................................................. 25 2.5.2 Biomarkers of effect ............................................................................... 26 Oxidative stress biomarkers ........................................................................ 26 Superoxide dismutase (SOD) ...................................................................... 26 Catalase (CAT) ........................................................................................... 26 Malondialdehyde (MDA) .............................................................................. 27 Protein carbonyls (PC) ................................................................................ 27 Reduced glutathione (GSH) ........................................................................ 27 2.5.3 RNA as a biomarker .............................................................................. 28 Chapter 3: Focus species ...................................................................................... 29 3. Introduction ....................................................................................................... 30 3.1 Taxonomy .................................................................................................... 31 3.2 External anatomy ......................................................................................... 32 3.3 Distribution ................................................................................................... 33 3.4 Feeding habits ............................................................................................. 34 ii | P a g e 3.5 Artificial spawning and its importance .......................................................... 34 Chapter 4 : Artificial breeding and embryonic development of Synodontis zambezensis (Peters, 1852) ................................................................................... 36 Abstract ................................................................................................................. 37 4. Introduction ....................................................................................................... 37 4.1 Materials and methods ................................................................................ 38 4.1.1 Water quality preferences ...................................................................... 39 4.1.2 Artificial breeding setup ......................................................................... 39 4.1.3 Aquaspawn® Injection, egg stripping and fertilisation ........................... 39 4.1.4 Egg tumbling and holding tank .............................................................. 40 4.1.5 Larvae, fry and fingerling maintenance .................................................. 41 4.1.6 Microscopic observation of egg development ........................................ 42 4.2 Results and discussion ...............................................................................
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