For Zebra Mussel (Dreissena Polymorpha) Control in Irish Waters
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Assessment and Utilisation of Zequanox® for Zebra Mussel (Dreissena polymorpha) Control in Irish Waters by Sara Meehan Submitted to the Institute of Technology, Sligo in fulfilment of the requirement for the degree of Doctor of Philosophy. October 2014 Supervisor: Dr. Frances Lucy, Institute of Technology, Sligo Declaration I declare that this thesis is my own work, and that it has not been previously submitted to any other institute or university. I give permission to the Yeats library, Institute of Technology, Sligo to lend this thesis on request. ------------------------------- Sara Meehan October, 2014 For Daniel “And seek not the depths of your knowledge with staff or sounding line. For self is a sea boundless and measureless” -Kahlil Gibran Acknowledgements Without the help and support of a number of individuals this thesis would not exist. I wish to acknowledge the following. My wonderful supervisor Dr. Frances Lucy who upon embarking on this journey told me she would support me all the way to the pulpit, well I am nearly there and have felt her encouragement and support throughout this whole process, for that I will be forever grateful. My parents Joan and Peter and my partner Marc, all the behind the scenes support and encouragement helped me to continue and stay strong. Also Marc and Dad your help with sampling deserves an extra thank you, so thanks lads! The McDonnell’s, my second family, thank you for making Killala a home away from home there have been many times over the course of this research when a home cooked meal from Finola has been just what I needed! To the Celbridge girls, having wonderful friends to spend weekends with at home and constantly feeling your love and support has made being away from home that much easier. The Mayo girls and one American, thanks for welcoming me to the fold! All the walks, chats and nights out were such a welcome distraction and without your friendship Ballina would not have felt like home. To Nana and Grandad and all my Donegal and Limerick family thanks for all the prayers and support. Patricia and Liz thank you both for always remembering me. Aine my soul sister, for all the visits to Mayo you were always a welcome distraction with guaranteed laughs, I will be forever grateful for your friendship. Remembering my dear friend Liz, who from an early age impressed upon me the value and importance of hard work in achieving results. You were an inspiration then and still are now. i Molly, thank you for all the yoga. It helped keep me sane and stress free throughout my research. My office buddies old and new, having a fun and relaxing work environment has certainly helped me be more productive, thanks Lorraine and Rory for all the laughs. Sheila, Alan, Carlos and Joe thank you for being so supportive and kind when I first began my research you are part of the reason I was successful. Marrone Bio Innovations for the research funding and in particular Sarahann and Bridget, thank you for all your assistance and support, my fondest memories of my research are of your visits. IT Sligo, for the research funding and special thanks to all the administrative and technical staff for all your assistance. TSGE, in particular Adam and Claire thank you for always being at the other end of the phone willing to answer all my questions. Inland Fisheries Ireland, Environmental Protection Agency, National Parks and Wildlife Service, Department of Agriculture Food and the Marine, Waterways Ireland and Sligo County Council thank you for supporting my research. ii Abstract Since the first arrival of the zebra mussel (Dreissena polymorpha) to Ireland in the early 1990’s, there has been a rapid secondary spread of these highly invasive mussels within many Irish freshwater systems suitable for their establishment. This has resulted in direct invasive impacts to infested lakes, rivers and canals, both to the freshwater biota and to man-made infrastructure. The availability of an effective, yet environmentally benign, mussel control product to replace those currently used, such as chlorine and other biocides is particularly needed to mitigate mussel fouling impacts in drinking water plants and other infested facilities. Zequanox® is a selective control product for dreissenids, i.e. zebra and quagga (D.rostriformis bugensis) mussels. It is a natural biocide, with the active ingredient being dead Pseudomonas fluorescens CL145A cells; a patented strain of a naturally occurring soil bacterium, which works by disrupting the mussels’ digestive system. This product was developed in the USA, and has been tested and used in a number of North American trials, with the aim of commercialising a cross-continental, effective and regulatory compliant control product. Research outputs from this thesis were utilised in the Zequanox regulatory application for commercial use within Europe. Ecotoxicology trials were carried out on nine species found commonly in Irish ecosystems. Results indicated that Zequanox does not negatively affect eight of these organisms at concentrations and treatment lengths required to get a >80% zebra mussel kill. Field trials were carried out to demonstrate the effectiveness of Zequanox in industry and open water and to monitor water quality during and after treatment. These included a biobox trial at a drinking water treatment plant and an open water trial in a canal. Both trials achieved high levels of zebra mussel mortality (up to 81%) and provided insights into practical application techniques. Additional laboratory assays were undertaken to determine the exact response relationship of juvenile zebra mussels to Zequanox. The results showed that juvenile mortality, on both experimental and control plates, are decreased with reduced handling, iii with subsequent recommendations given for future juvenile treatment and counting procedures in the field. After the two field trials (drinking water treatment plant and canal) were undertaken, it was apparent that mortality rates after treatment were lower in Ireland and also took longer to occur than similar trials carried out in North America. It was suspected that the higher water temperatures during North American trials (>25C) meant that the zebra mussels were more metabolically active and therefore ingested more product. A number of comparative studies commenced to evaluate the effect of Zequanox on North American zebra mussels versus European zebra mussels. The results of this study showed that under the same temperature regimes mortality is similar. North American mussels were found to ingest more product in the initial eight hours, however by 24 hours, product concentration was similarly low for both mussel groups. The results of this trial allowed for industry recommendations to be made regarding the timing of treatments in Europe. In conclusion this research has bridged the gap between the use of Zequanox in Europe and North America, showing there is potential for Zequanox to control zebra mussels in Europe not only in industry but also in open water. This study has also demonstrated Zequanox’s potential to replace chlorine as the traditionally used control method, thereby reducing the environmental impact of mussel control on freshwater ecosystems. iv Contents Chapter 1 Introduction 1.1 Overview of Chapter 1 1.2 Introduction to Invasive Species 1 1.2.1 European and Irish Legislation Relevant to Invasive Species 2 1.3 The Zebra Mussel 3 1.3.1 Introduction to the Zebra Mussel 3 1.3.2 Zebra Mussel Reproduction 4 1.3.3 Feeding in Zebra Mussels 6 1.3.4 Environmental Requirements 8 1.4 Impacts of Zebra Mussels 10 1.4.1 Introduction 10 1.4.2 Impacts to the Benthic Environment 11 1.4.3 Impacts to Fish 11 1.4.4 Impacts to Water Clarity 12 1.4.5 Impacts to Elements and Compounds 13 1.4.6 Impacts to Unionids 14 1.4.7 Impacts to Industry 15 1.5 Control of Zebra Mussels 16 1.5.1 Introduction 16 1.5.2 Temperature 16 1.5.3 Chlorine 17 v 1.5.4 Other Chemicals 18 1.5.5 UV Light 19 1.5.6 Microbial Biofilm 19 1.5.7 Natural Antifouling Agents 20 1.5.8 Combinations 20 1.5.9 BioBullets 21 1.5.10 Zequanox 21 1.6 Zebra Mussel Monitoring 23 1.6.1 Introduction 23 1.6.2 Settlement Plates 25 1.6.3 Sidestream Monitoring 26 1.6.4 Mesocosm 27 1.7 Ecotoxicology Testing 27 1.7.1 Rational for Selecting Organisms for Ecotoxicology Trials 28 1.8 Aims and Objectives of Thesis 34 1.9 Tasks and Methods 35 Chapter 2 The impact of Zequanox® on Selected Non Target Irish Aquatic Organisms 2.1 Introduction to paper 37 2.2 Introduction 37 2.2.1 Guidelines for Ecotoxicology Testing 38 2.3 Objectives 39 vi 2.4 Materials and Methods 39 2.4.1 General Procedures 40 2.4.2 Test Organism Origin 43 2.4.3 Dilution water 45 2.4.4 Determination of Test Concentrations 46 2.4.5 Preparation of Test Concentrations 49 2.4.6 Test Conditions 50 2.4.7 Test Procedures 53 2.4.8 Validity of the Test 54 2.5 Results and Discussion 55 2.5.1 Water Quality 55 2.5.1.1 Water Quality Parameters (Raw Data Comparisons) 55 2.5.1.2 Statistical Analysis of Water Quality (Two-Way ANOVA) 57 2.5.2 Ecotoxicology 63 2.5.2.1 Chironomus 63 2.5.2.2 Asellus aquaticus 68 2.5.2.3 Ephemerella ignita 70 2.5.2.4 Mytilus edulis 73 2.5.2.5 Anodonta 75 2.5.2.6 Daphnia pulex 76 2.5.2.7 Austropotamobius pallipes 78 2.5.2.8 Lymnaea peregra 80 2.5.2.9 Salmo trutta (parr) 82 2.6 Overall Conclusions 84 vii Chapter 3 Comparing a Microbial Biocide and Chlorine as Zebra Mussel Control Strategies in an Irish Drinking Water Treatment Plant 3.1 Introduction to Paper 89 3.2 Introduction 90 3.2.1 Sligo Drinking Water Treatment Plant, Ireland