Across Latitudes: Using Molluscs as Biomonitors to Assess Spatial and Temporal Distribution of Tributyltin (TBT) in Eastern Canada and the British Virgin Islands by Cassander Patrice Titley-O'Neal MSc. Applied Marine Science, University of Plymouth, 2001 BSc. Marine Biology, University of North Carolina at Wilmington, 1999 A DISSERTATION SUBMITTED IN PARTIAL FULFILLMENT OF THE REQUIREMENTS FOR THE DEGREE OF Doctor of Philosophy In the Graduate Academic Unit of Biology Supervisor: Bruce A. MacDonald, Ph.D, Department of Biology, Saint John Examining Board: Lucy Wilson, Ph.D, Department of Geology, Saint John (Chair) Heather L. Hunt, Ph. D, Department of Biology, Saint John James D. Kieffer, Ph.D, Department of Biology, Saint John External Examiner: Adrienne J. Bartlett, Ph. D, Environment Canada, Centre for Inland Waters This dissertation is accepted Dean of Graduate Studies THE UNIVERSITY OF NEW BRUNSWICK January 2011 ©Cassander P. 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Canada f Press toward the mark (Phillipians 3:14) DEDICATION This PhD thesis is dedicated to the two persons that mean the world to me. They are the ones who gave me life, who believed in me even when I didn't believe in myself, who gave me the strength to push on during the dark days, and who have instilled in me everything that I am today. In memory of my Dad (Amos Jerome O'Neal a.k.a Romey): Your untimely passing on November 27th, 2009 during the last stages of my PhD thesis rocked me to the core. I miss our late night chats on the telephone until the wee hours of the morning talking about life, fishing or whatever was important at the time. You always use to tell me, "Sandra... Some birds weren't meant to be in cages and you are one of them." I reflect on that often. It is because of you I pushed until the end even when I felt like giving up. I hope that with that bird's eye view from Heaven you can see that your little girl has made it! You will be in my heart FOREVER. R.I.P. in Smdm z ii To my Mom (Barbara Hyacinth Titley): It is because of you that I am even pursuing my PhD. You continually nagged me until I sent in the application for the scholarship even though I did not feel like applying. Thank you for your foresight. What can I say to the woman who has been and continues to be an unbelievable source of strength emotionally and psychologically? You are my strength, my strong tower, the wind beneath my wings. During every test and trial you would remind me, "Remember who you are in Christ!" Hopefully you will be able to rest easier and know that even though my world has turned upside down, and in spite of all the fiery darts, hurdles and speed bumps, both academically and personally, I have made it to the end of this PhD journey! •Sano iii ABSTRACT Three species of molluscs (Nucella lapillus, Mytilus edulis and Strombus gigas) were used as biomonitors to evaluate tributyltin (TBT) pollution spatially and temporally in two Maritime Provinces in Atlantic Canada and the British Virgin Islands (BVI) during the summers of 2007 to 2009. In addition, a review of the literature on the effects of organotins on female gastropods from 1962 to 2009 was undertaken in an effort to summarize how the terminology, science and theories have evolved over the last 40 plus years. In Canada, data confirm that TBT was bioavailable based on chemical analysis of samples collected at locations within harbour sites, and TBT levels were high enough to induce imposex, the superimposition of a penis and/or vas deferens, in N. lapillus at harbour sites where they were found. A comparison of the current study with previous surveys in the same geographic areas implies that the 1989 Canadian regulations on TBT have been effective in reducing TBT levels in sediment and imposex severity in N. lapillus in Saint John Harbour and Halifax Harbour. Conversely, during the same time period, TBT levels in sediment from Sydney Habour increased. Butyltins were also detected in sediment from reference sites where vessels < 25 m in length were docked at marinas and fishing harbours. In the BVI, data from this study verify that TBT is bioavailable based on the incidence of imposex in female S. gigas collected from areas adjacent to boating activity e.g. shipyards, marinas and harbours, and the presence of TBT and its degradative iv product, dibutyltin (DBT), in S. gigas, turtle grass (Thalassia testidium) and marine algae from corresponding sites. To date, female masculinisation (FM) of gastropods has been documented in > 260 species globally. Although most studies attribute FM in gastropods to TBT, there is evidence that "pseudo-imposex" occurred prior to TBT's use as a biocide in antifouling paint in the 1960s, "natural imposex" occurs, and there are other compounds besides TBT that can cause FM. Even after 40+ years of research, there is no consensus on how the actual mechanism of induction occurs, although five theories have been proposed. v ACKNOWLEDGEMENTS I would first of all like to give all honour, praise and thanks to God for without him this PhD would not have been attainable. No man is an island and the achievement of my PhD would not have been possible without financial support, colleagues, and the love and support of family and friends. At this time I would also like to recognise all the financial stakeholders that have contributed towards my PhD for without them the wheels of research would have ceased before I even got started: the Canadian Bureau for International Education (CBIE), the Natural Sciences and Engineering Research Council (NSERC) of Canada (Dr. Bruce MacDonald), Canadian Research Chair in Molecular Ecotoxicology (Dr. Emilien Pelletier), H. Lavity Stoutt Community College (HLSCC), Hon. Ralph T. O'Neal - Premier of the Virgin Islands, Ministry of Education and Culture, University of New Brunswick, Hon. Irene Penn-O'Neal, J. S. Little International Study Fellowship, and Society of Environmental Toxicology and Chemistry (SETAC). First and foremost, I must say sincerest appreciation to my supervisor, Dr. Bruce MacDonald, who allowed me to spread my wings and fly with my project. Thanks for the confidence, encouragement and numerous life lessons taught along the way. As we would say in the Virgin Islands, "Meh son I couldn'ta ask for a better supervisor. Big up!" I would also like to recogonise my supervisory committee, Drs. Karen Kidd, Deb MacLatchy and Kelly Munkittrick, for their invaluable insight, critiques and suggestions throughout the development and completion of my thesis. I say kudos and a special vi thank you to Kelly for his foresight, vigilance, and assistance with the development and production of the review article. To my French collaborators, Drs. Emilien Pelletier and Richard Saint-Louis, at the Institut des sciences de la mer de Rimouski (ISMER) at the Universite du Quebec a Rimouski, aka "Remoteski", merci beaucoup for the confidence, support and taking me on as a "stagiaire". Thanks to my field crew in Canada for all your assistance with collecting sediment, mussels and dogwhelks from the various sites throughout New Brunswick and Nova Scotia: my boat captains, Dr. Bruce Hatcher and Mr. Wayne Armstrong, and field assistants, Erin Holtz, Adrian Hamer and Matthew Hatcher. I would also like to thank the other arm of my field crew in the British Virgin Islands for their assistance with collecting samples there: Orville Phillip, Lenette Lewis, Geoff Brooks and my stand behind me captain Reynold "Al" Frazer. I would like to recognise Adrian Hamer for his invaluable assistance with the production of the review article, and I'm sure that by now you know more than you would ever want to know about imposex, intersex and female penes to last you a life time.