by Wong On Nei. / Thesis submitted in: October 2004. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2005. / Includes bibliographical references (leaves 101-116). / Abstracts in English and Chinese. / ABSTRACT --- p.ii / 摘要 --- p.iv / ACKNOWLEDGEMENTS --- p.vi / TABLE OF CONTENTS --- p.vii / LIST OF TABLES --- p.xi / LIST OF FIGURES --- p.xiii / Chapter CHAPTER ONE --- INTRODUCTION --- p.1 / Chapter 1.1 --- Background --- p.1 / Chapter 1.2 --- Lethal and sublethal effects of SS on fish --- p.2 / Chapter 1.2.1 --- Biological effects --- p.2 / Chapter 1.2.2 --- Molecular biomarkers --- p.3 / Chapter 1.2.2.1 --- Acetylcholinesterase activity inhibition assay --- p.4 / Chapter 1.2.2.2 --- Induction of cytochrome P450 mRNA --- p.5 / Chapter 1.2.2.3 --- Induction of metallothionein mRNA --- p.6 / Chapter 1.2.3 --- Study on CYP1A and MT expression / induction --- p.8 / Chapter 1.2.3.1 --- Reverse Transcription (RT) --- p.8 / Chapter 1.2.3.2 --- Polymerase chain reaction (PCR) --- p.8 / Chapter 1.3 --- Objectives --- p.11 / Chapter CHAPTER TWO --- MATERIALS AND METHODS --- p.13 / Chapter 2.1 --- Study sites --- p.13 / Chapter 2.2 --- Sediment --- p.15 / Chapter 2.2.1 --- Sediment samples collection --- p.15 / Chapter 2.2.2 --- Sediment handling --- p.15 / Chapter 2.2.3 --- Sediment dry-wet (w/w) ratio measurement --- p.15 / Chapter 2.2.4 --- Heavy metal content analysis --- p.16 / Chapter 2.2.5 --- Organic content analysis --- p.16 / Chapter 2.3 --- Test organism --- p.17 / Chapter 2.4 --- Bioassays --- p.17 / Chapter 2.4.1 --- 10-day exposure treatments --- p.17 / Chapter 2.4.1.1 --- Experimental setup --- p.17 / Chapter 2.4.1.2 --- Procedure --- p.21 / Chapter 2.4.1.3 --- Tissue sample collection --- p.22 / Chapter 2.4.2 --- 30-day exposure treatments --- p.22 / Chapter 2.4.2.1 --- Behavioural observations --- p.23 / Chapter 2.4.2.2 --- Tissue sample collection --- p.23 / Chapter 2.5 --- Molecular Biomarkers --- p.25 / Chapter 2.5.1 --- Tested samples --- p.25 / Chapter 2.5.2 --- Acetylcholinesterase activity inhibition assay --- p.25 / Chapter 2.5.2.1 --- Acetylcholinesterase activity assay --- p.25 / Chapter 2.5.2.2 --- BioRad Bradford assay --- p.26 / Chapter 2.5.2.3 --- Calculation of specific enzyme activity --- p.26 / Chapter 2.5.3 --- Study on CYP1A and MT expression / induction --- p.27 / Chapter 2.5.3.1 --- Gill and liver tissue samples --- p.27 / Chapter 2.5.3.2 --- Preparation of ribonuclease free reagents and apparatus --- p.27 / Chapter 2.5.3.3 --- Isolation of total RNA --- p.27 / Chapter 2.5.3.4 --- Spectrophotometric analyses of DNA and RNA --- p.28 / Chapter 2.5.3.5 --- First strand cDNA synthesis --- p.28 / Chapter 2.5.3.6 --- Cloning and sequencing of CYP1A and MT gene --- p.29 / Chapter 2.5.3.7 --- RT-PCR co-amplification of CYP1A and 18S rRNA --- p.34 / Chapter 2.5.3.8 --- Real-time RT-PCR --- p.36 / Chapter CHAPTER THREE --- RESULTS --- p.39 / Chapter 3.1 --- Sediment chemistry --- p.39 / Chapter 3.1.1 --- Sediment dry-wet (w/w) ratio --- p.39 / Chapter 3.1.2 --- Heavy metal content of sediments --- p.39 / Chapter 3.1.3 --- Levels of total PCBs and PAHs in sediment --- p.39 / Chapter 3.2 --- Monitoring of test conditions --- p.42 / Chapter 3.3 --- Bioassays --- p.42 / Chapter 3.3.1 --- Survivorship --- p.42 / Chapter 3.3.2 --- Growth --- p.46 / Chapter 3.3.3 --- Feeding rate --- p.51 / Chapter 3.3.4 --- Behaviour --- p.54 / Chapter 3.3.5 --- Sediment clogging --- p.59 / Chapter 3.3.6 --- Body lesions --- p.59 / Chapter 3.3.7 --- Abnormal behaviour --- p.59 / Chapter 3.4 --- Molecular biomarkers --- p.63 / Chapter 3.4.1 --- Acetylcholinesterase activity inhibition assay --- p.63 / Chapter 3.4.2 --- Cloning and sequencing of CYP1A and MT gene --- p.66 / Chapter 3.4.3 --- RT-PCR co-amplification of CYP1A and 18S rRNA --- p.73 / Chapter 3.4.4 --- Real-time RT-PCR --- p.77 / Chapter CHAPTER FOUR --- DISCUSSION --- p.84 / Chapter 4.1 --- Sediment chemistry --- p.84 / Chapter 4.2 --- Biological responses --- p.85 / Chapter 4.3 --- Molecular biomarkers --- p.91 / Chapter 4.3.1 --- Acetylcholinesterase activity inhibition assay --- p.91 / Chapter 4.3.2 --- Cloning and sequencing of CYP1A and MT gene --- p.93 / Chapter 4.3.3 --- RT-PCR co-amplification of CYP1A and 18S rRNA --- p.93 / Chapter 4.3.4 --- Real-time RT-PCR --- p.95 / Chapter 4.4 --- Recommendations --- p.99 / Chapter 4.5 --- Conclusions --- p.100 / REFERENCES --- p.101 / APPENDIX --- p.117