Imposex in Marine Gastropods from the Atlantic Coast of South Africa
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Imposex in marine gastropods from the Atlantic coast of South Africa N Roos 21751412 Dissertation submitted in fulfilment of the requirements for the degree Magister Scientiae in Environmental Sciences at the Potchefstroom Campus of the North-West University Supervisor: Prof H Bouwman Co-supervisor: Dr A Averbuj April 2016 Imposex in marine gastropods from the Atlantic coast of South Africa DECLARATION I declare that “Imposex in marine gastropods from the Atlantic coast of South Africa” to be my own work, that is has not been previously submitted for any degree or examination purposes at this or any other university, and that all the sources used or quoted, have been indicated and acknowledged. Signed : Nicole Roos Date: 29 April 2016 ii Imposex in marine gastropods from the Atlantic coast of South Africa DEDICATION AND ACKNOWLEDGEMENTS First and foremost, I would like to thank my Holy Father and Saviour, Jesus Christ. Through His grace and mercy, I have been able to achieve that which I never thought possible. I would also like to thank the following people and institutions: • My thesis advisor, Prof Henk Bouwman from the North West University. Prof Bouwman encourgaed me to take charge of this study and make my own decisions. When I veered off the path, Prof was always there to steer me in the right direction. • My co-supervisor, Dr Andres Averbuj, from LARBIM-IBIOMAR-CONICET (Puerto Madryn, Argentina) for his expert guidance and help with this project. • Simone Roos for graphical and technical editing. • Martin van Rensburg for stastistical analysis. • My fellow students for their assitance in the field and laboratory: JP ‘Carlos’ Huizeman, Chani Swanepoel, Veronica van der Schyff and Marinus du Preez. • The South African National Parks board and CapeNature for permitting me to sample gastropods from their delicate ecosystems. • Lastly, but definitely not least I would like to express my utmost gratitude to my parents, partner and family for providing me with continous support and encouragement throughout my years of study and through the process of research and completing this thesis. This would not have been possible without you. Thank you. iii Imposex in marine gastropods from the Atlantic coast of South Africa ABSTRACT Tributyltin (TBT) is an organotin compound used as an antifouling agent in paint that is applied to the hulls of vessels to prevent the formation of biofilms. This highly toxic antifouling compound leaches into the water and has subsequent negative effects on marine life, especially marine gastropods. One of the most serious effects of exposure to TBT is the growth of a penis/vas deferens in female gastropods; this phenomenon is referred to as imposex (imposed sex). Marine gastropods are therefore useful bioindicators as imposex is related to TBT exposure. Imposex may affect the reproduction of these organisms and thus affect coastal ecology. The consumption of TBT-contaminated foodstuffs may also pose a risk to human health. The use of antifouling paints containing organotin compounds such as TBT continues in developing countries, despite the fact that the International Maritime Organization (IMO) banned the global use of organotin compounds in antifouling paints applied to ships in 2008, by country Parties electing to do so. This is the first report of an imposex-sediment survey used to evaluate TBT contamination along South Africa’s Atlantic coastline. This was achieved by sampling caenogastropods from sites presumed impacted and non-impacted by TBT pollution and by using various biological indices: Percentage Imposex (% I), Relative Penis Length Index (RPLI), Relative Penis Size Index (RPSI), and Male:Female ratio (M:F). TBT and DBT (dibutyltin) concentrations were measured in sediments to provide additional information regarding TBT pollution as gastropods usually inhabit these sediments and imposex may be related to these concentrations. This study showed that imposex is prevalent along the coastline and high concentrations of TBT have been measured, which is of major concern. Results have shown that TBT and DBT levels are related to boating activity, including imposex prevalence. Research will make valuable information available to necessary parties so that regulations and protective measures may be enforced to protect our marine life. Key words: TBT (Tributyltin); Imposex; Caenogastropods; Biomonitoring; RPLI; RPSI; M:F ratio; Organotin compounds (OTCs); Endocrine disruption; Marine sediments. iv Imposex in marine gastropods from the Atlantic coast of South Africa TABLE OF CONTENTS DECLARATION ........................................................................................................... II DEDICATION AND ACKNOWLEDGEMENTS ........................................................... III ABSTRACT ................................................................................................................. IV TABLE OF CONTENTS ............................................................................................... V LIST OF FIGURES .................................................................................................... VIII LIST OF TABLES ....................................................................................................... IX LIST OF ACRONYMS .................................................................................................. X CHAPTER 1: INTRODUCTION ................................................................................... 1 CHAPTER 2: LITERATURE REVIEW ......................................................................... 6 2.1 FOULING AND ANTIFOULING ................................................................................. 6 2.2 ORGANOTIN COMPOUNDS AND TBT ..................................................................... 7 2.2.1 Properties ..................................................................................................... 7 2.2.2 History of TBT .............................................................................................. 8 2.2.3 Effects and toxicity ....................................................................................... 9 2.2.4 Sources ...................................................................................................... 12 2.2.5 Routes of exposure .................................................................................... 13 2.2.6 TBT in the environment .............................................................................. 14 2.2.7 Other possible pollutants ............................................................................ 18 2.3 ENDOCRINE DISRUPTION .................................................................................. 19 2.3.1 TBT as an endocrine disruptor ................................................................... 22 2.4 GASTROPODS .................................................................................................. 23 2.4.1 The gastropod reproductive system ........................................................... 25 2.4.2 The gastropod endocrine system ............................................................... 26 2.5 IMPOSEX ......................................................................................................... 28 2.5.1 Imposex indices ......................................................................................... 34 v Imposex in marine gastropods from the Atlantic coast of South Africa 2.5.2 Pathway of TBT-induced imposex.............................................................. 36 2.6 BIOMONITORING AND GASTROPODS ................................................................... 37 2.7 ALTERNATIVES TO TBT AS ANTIFOULANTS ......................................................... 41 CHAPTER 3: MATERIALS AND METHODS ............................................................ 46 3.1 SPECIES SELECTION ......................................................................................... 46 3.2 SITE SELECTION ............................................................................................... 46 3.3 SITE DESCRIPTIONS ......................................................................................... 46 3.4 GASTROPOD DESCRIPTION ............................................................................... 60 3.5 GASTROPOD COLLECTION ................................................................................ 65 3.6 SEDIMENT COLLECTION .................................................................................... 66 3.7 MEASUREMENTS .............................................................................................. 66 3.8 IMPOSEX ANALYSIS .......................................................................................... 73 3.9 CHEMICAL ANALYSIS ........................................................................................ 74 3.10 STATISTICAL ANALYSIS ..................................................................................... 74 CHAPTER 4: RESULTS ............................................................................................ 75 4.1 TBT/DBT CONCENTRATIONS IN SEDIMENTS ....................................................... 75 4.1.1 TBT/DBT inputs.......................................................................................... 77 4.1.2 Boating Activity vs. DBT (µg/kg) ................................................................. 77 4.1.3 Boating Activity vs. TBT (µg/kg) ................................................................. 80 4.2.