Ryerson University Digital Commons @ Ryerson Theses and dissertations 1-1-2013 Toxicity Assessment Of The Antimicrobial Triclocarban Using Sub-Lethal Behaviour And Reproduction Endpoints Melanie Raby Ryerson University Follow this and additional works at: http://digitalcommons.ryerson.ca/dissertations Part of the Environmental Sciences Commons Recommended Citation Raby, Melanie, "Toxicity Assessment Of The Antimicrobial Triclocarban Using Sub-Lethal Behaviour And Reproduction Endpoints" (2013). Theses and dissertations. Paper 2017. This Thesis is brought to you for free and open access by Digital Commons @ Ryerson. It has been accepted for inclusion in Theses and dissertations by an authorized administrator of Digital Commons @ Ryerson. For more information, please contact [email protected]. TOXICITY ASSESSMENT OF THE ANTIMICROBIAL TRICLOCARBAN USING SUB-LETHAL BEHAVIOUR AND REPRODUCTION ENDPOINTS by Melanie Raby Bachelor of Science (Honours) University of Waterloo, 2011 A thesis presented to Ryerson University in partial fulfillment of the requirement for the degree of Master of Applied Science in the Program of Environmental Applied Science and Management Toronto, Ontario, Canada, 2013 © MELANIE RABY, 2013 Author’s Declaration I hereby declare that I am the sole author of this thesis. This is a true copy of the thesis, including any required final revisions, and accepted by my examiners. I authorize Ryerson University to lend this thesis to other institutions or individuals for the purpose of scholarly research. I further authorize Ryerson University to reproduce this thesis by photocopying or by other means, in total or in part, at the request of other institutions or individuals for the purpose of scholarly research. I understand that my thesis may be made electronically available to the public. ii Abstract Toxicity assessment of the antimicrobial triclocarban using sub-lethal behaviour and reproduction endpoints Master of Applied Science, 2013 Melanie Raby Environmental Applied Science and Management Ryerson University Aquatic environments have long been used as disposal sites for domestic and industrial wastes, resulting in increasing chemical contamination, decreased water quality, and concern for ecosystem health and drinking water sources. This study utilized bioassays, the “golden standard” method to measure biological impact, to assess the toxicity of the widely found surface water contaminant, the antimicrobial triclocarban. Culturing protocols were implemented to provide healthy, age-synchronized organisms for bioassays. Behaviour and reproduction were demonstrated as useful endpoints while refining these methods using 4-chlorophenol and were successfully implemented in the toxicity assessment of triclocarban. While no sub-lethal behavioural impact was seen, 10.0 ppb triclocarban was found to delay reproduction in Daphnia magna. This delay could result in population, community, and ecosystem-level responses. iii Acknowledgements This project would not have been possible without the guidance, encouragement, and input of so many people. First, I would like to thank Dr. Lynda McCarthy who saw an ecotoxicologist in a naive biochemist. Thank you for the opportunity and encouragement throughout this journey. Thank you to Karen Puddephatt for teaching me the ropes, for your friendship and for being my knitting buddy. To Dr. Jorge Loyo, my partner in crime, thank you for helping me with “the water problem” and most importantly for your friendship and support. I must also extend my gratitude to fellow graduate students Mark Tiley and Daniel Johnson and research assistants Alicia Falls and Laura Taylor for their friendship and help in the lab. Additionally, I would like to thank Dave Poirier and Richard Chong-Kit of the MOE for their invaluable advice and their (many) gifts of Lumbriculus and Chlorella, along with Dr. Warren Norwood of CCIW for his helpful Hyalella azteca culturing advice. I would also like to thank Dr. Andrew Laursen for his guidance with my data analysis. Finally, I would like to thank Dr. Michal Bardecki, Dr. Stephen Wylie, and Dr. Gideon Wolfaardt for their roles as examiners on my defence committee. I would like to thank my parents, Laura and Rob, my sisters, Lisa and Christine, my grandmother Clara, as well as the Staples family for their love and support throughout this journey. Finally, a special thank you to my partner Robbie for his patience, encouragement and love. iv Table of Contents 1.0 INTRODUCTION ........................................................................................................................ 1 1.1 Overview ...................................................................................................................................... 1 1.2 Objectives ..................................................................................................................................... 2 1.3 Importance of Bioassays............................................................................................................... 2 1.3.1 Review of Bioassay Organisms in WWTP Effluent Toxicity Research ............................... 3 1.3.2 Single-Species Bioassays ...................................................................................................... 4 1.4 Protocol Development .................................................................................................................. 4 1.4.1 Reference Toxicants .............................................................................................................. 5 1.5 Bioassay Endpoints ...................................................................................................................... 9 1.5.1 Rationale for Selection of Test Endpoints ............................................................................. 9 1.5.2 Behaviour ............................................................................................................................ 10 1.5.3 Reproduction ....................................................................................................................... 19 1.6 Bioassay Organisms ................................................................................................................... 19 1.6.1 Rationale for Selection of Bioassay Organisms .................................................................. 19 1.6.2 Daphnia magna ................................................................................................................... 20 1.6.3 Hyalella azteca .................................................................................................................... 27 1.6.4 Lumbriculus variegatus ....................................................................................................... 33 1.6.5 Lemna minor ........................................................................................................................ 37 1.7 Triclocarban ................................................................................................................................ 41 1.7.1 Environmental Impact of Triclocarban ................................................................................ 42 2.0 METHODS ..................................................................................................................................... 45 2.1 General ....................................................................................................................................... 45 2.1.1 Dechlorination of Municipal Drinking Water ..................................................................... 45 2.1.2 Glassware Washing Procedure ............................................................................................ 46 2.1.3 General Bioassay Conditions............................................................................................... 46 2.2 Culturing and Age Synchronization ........................................................................................... 46 2.2.1 Daphnia magna ................................................................................................................... 47 2.2.2 Hyalella azteca .................................................................................................................... 52 2.2.3 Lumbriculus variegatus ....................................................................................................... 56 2.2.4 Lemna minor ........................................................................................................................ 58 2.3 Positive Toxicant ........................................................................................................................ 60 v 2.3.1 Behaviour Development ...................................................................................................... 60 2.3.2 Behaviour ............................................................................................................................ 62 2.3.3 Life Cycle ............................................................................................................................ 63 2.4 Triclocarban ................................................................................................................................ 65 2.4.1 Behaviour ............................................................................................................................ 65 2.4.2 Life Cycle ...........................................................................................................................