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Feeding Behaviour of Lumbriculus Variegatus As an Ecological Indicator of in Situ Sediment Contamination

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Feeding Behaviour of Lumbriculus Variegatus As an Ecological Indicator of in Situ Sediment Contamination Feeding behaviour of Lumbriculus variegatus as an ecological indicator of in situ sediment contamination Thesis submitted to the University of Stirling for the degree of Doctor of Philosophy by Philip Mark Williams Institute of Aquaculture January 2005 DECLARATION This thesis has been composed in its entirety by the candidate and no part of this work has been submitted for any other degree Candidate : Philip Williams 2 Abstract Previous studies have demonstrated that the feeding behaviour of Lumbriculus variegatus may be significantly inhibited during exposure to toxic substances. The potential use of an in situ sediment bioassay, using L.variegatus post-exposure feeding inhibition as an endpoint, was investigated. The bioassay consisted of exposing animals in the field for a six-day exposure period and feeding rates were measured immediately afterwards over a twenty-four hour post-exposure period. The bioassay methodology developed in the laboratory produced a consistent baseline response that was reliable and repeatable. Endpoint sensitivity was demonstrated under laboratory conditions, where bioassay organisms exhibited delayed recovery from feeding inhibition after previous exposure to sediment-associated contaminants. The apparent insensitivity of the bioassay to sediment- associated metals means that the technique should only be used as part of a suite of bioassays that employ representative deposit feeders. The ecological relevance of the bioassay endpoint was also demonstrated by comparing short-term measures of post- exposure feeding inhibition with the longer-term effects of a toxicant on L.variegatus populations. The bioassay methodology was successfully adapted for in situ use. Post-exposure feeding inhibition was detected at contaminated field sites. However, the consistent baseline response produced in the laboratory could not be replicated during deployments of the bioassay at upstream (“clean”) field sites. Increased environmental “noise” may have 3 been a result of a number of confounding factors that could limit the sensitivity of the bioassay endpoint if not adequately controlled. Despite the above concerns, the in situ bioassay is suggested to represent a useful tool, which uses a more realistic field exposure scenario to investigate the effects of sediment-associated toxicants with an important functional component of aquatic ecosystems. 4 Acknowledgements Firstly I would like to thank my supervisors Drs.Donald Baird and Nigel Wilby for their advice and support during the completion of this work and for critically reviewing this thesis. Special thanks also go to Matti Leppanen and Anne Conrad for helping me get things started, Cori Critchlow-Watton for spending time reviewing this thesis, Billy Struthers for his help with chemical analysis, Carlos Barata and Kate Howie for advice on statistics, and finally Brian Howie, Charlie Harrower, Jane Lewis and Betty Stenhouse for their help obtaining the equipment required for this work. Thanks to the staff and students at the University of New Brunswick (Fredericton and Saint John) and National Water Research Institute (UNB, Fredericton) who made my visit very enjoyable and productive. In particular Sandra Brasfield, Michelle Gray, Genvieve Vallieres, Marni Turnbul, Kristi Heard, Joseph Culp and Eric Luiker. Last, but not least, to Ruth, my family and friends for their support and encouragement throughout. 5 CONTENTS Abstract…………………………………………………………………………………….3 Acknowledgements………………………………………………………………………...5 List of Tables……………………………………………………………………………...15 List of Figures…………………………………………………………………………….18 1 CHAPTER 1 - INTRODUCTION .............................................................................23 1.1 GENERAL INTRODUCTION..............................................................................23 1.2 ESTABLISHED SEDIMENT ASSESSMENT TECHNIQUES...........................24 1.2.1 Chemistry-based assessments and sediment quality values ...........................24 1.2.2 Macro-invertebrate community analysis ........................................................26 1.2.3 Laboratory-based invertebrate bioassays........................................................28 1.2.4 Integrated “weight of evidence” approaches ..................................................30 1.3 FIELD (IN SITU) BIOASSAYS ............................................................................31 1.3.1 Pros and cons with in situ bioassays for sediment assessment.......................32 1.3.2 Single species in situ bioassays and sub-lethal endpoints ..............................34 1.3.3 Feeding inhibition as a bioassay endpoint ......................................................35 1.3.4 Feeding inhibition as an in situ bioassay endpoint .........................................37 1.4 SELECTION OF FEEDING INHIBITION OF LUMBRICULUS VARIEGATUS AS AN IN SITU BIOASSAY ENDPOINT ......................................................................38 6 1.4.1 Basic ecology..................................................................................................38 1.4.2 Ecological relevance of L.variegatus feeding inhibition................................40 1.4.3 Practical considerations with L.variegatus .....................................................41 1.4.4 Measuring the feeding behaviour of L.variegatus in situ ...............................42 1.5 BIOASSAY DEVELOPMENT CRITERIA..........................................................43 1.5.1 Bioassay relevance..........................................................................................43 1.5.2 Bioassay reliability.........................................................................................44 1.5.3 Cost and timing...............................................................................................45 1.6 THESIS AIMS .......................................................................................................45 2 CHAPTER 2 - GENERAL METHODS AND BIOASSAY DEVELOPMENT.....47 2.1 INTRODUCTION: GENERAL METHODS ........................................................47 2.2 AIMS: GENERAL METHODS.............................................................................48 2.3 MATERIALS AND METHODS: GENERAL METHODS..................................49 2.3.1 Maintenance of L. variegatus cultures............................................................49 2.3.1.1 Stirling 2002-2003 (Chapters 2 and 3) ....................................................49 2.3.1.2 New Brunswick 2003 (Chapter 4)...........................................................50 2.3.1.3 Stirling 2003-2004 (Chapter 5) ...............................................................51 2.3.2 Water quality analysis of cultures...................................................................52 2.3.2.1 Stirling 2002-2003 (Chapters 2 and 3) and 2003-2004 (Chapter 5)........52 2.3.2.2 New Brunswick 2003 (Chapter 4)...........................................................53 2.3.3 Collection of field sediments ..........................................................................54 2.3.3.1 Stirling 2002-2003 (Chapters 2 and 3) ....................................................54 2.3.3.2 Stirling 2003-2004 (Chapter 5) ...............................................................57 7 2.3.3.3 New Brunswick 2003 (Chapter 4)...........................................................57 2.4 INTRODUCTION: BIOASSAY DEVELOPMENT.............................................59 2.4.1 Factors influencing the feeding rate of L.variegatus ......................................60 2.4.1.1 Reproduction ...........................................................................................60 2.4.1.2 Worm size................................................................................................62 2.4.1.3 Population density...................................................................................62 2.4.1.4 Temperature.............................................................................................63 2.4.1.5 Additional considerations........................................................................64 2.5 AIMS: BIOASSAY DEVELOPMENT .................................................................65 2.6 MATERIALS AND METHODS: BIOASSAY DEVELOPMENT ......................65 2.6.1 Experiment 1- Controlling reproduction and worm size versus egestion rate65 2.6.1.1 Set-up and identification of non-feeding worms .....................................66 2.6.1.2 Measurement of L.variegatus egestion rate.............................................68 2.6.2 Experiment 2 – Number of animals per replicate and length of the post- exposure feeding period ................................................................................................70 2.6.3 Experiment 3 – The effects of exposure temperature on post-exposure egestion rate...................................................................................................................72 2.6.4 Experiment 4 - Statistical power of the proposed bioassay methodology......74 2.7 RESULTS ..............................................................................................................76 2.7.1 Experiment 1...................................................................................................76 2.7.2 Experiment 2...................................................................................................79 2.7.3 Experiment 3...................................................................................................80
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