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Cyanobacterial Monitoring & Cyanotoxin Analysis ÅBO AKADEMIS FÖRLAG – ÅBO AKADEMI UNIVERSITY PRESS TOXIC Cyanobacterial Monitoring and Cyanotoxin Analysis Jussi Meriluoto and Geoffrey A. Codd Editors ACTA ACADEMIAE ABOENSIS, SER. B Mathematica et physica Mathematics, science, engineering Edited by Göran Högnäs Vol. 65, no. 1 TOXIC TOXIC Cyanobacterial Monitoring and Cyanotoxin Analysis Jussi Meriluoto and Geoffrey A. Codd Editors ÅBO 2005 ÅBO AKADEMIS FÖRLAG – ÅBO AKADEMI UNIVERSITY PRESS CIP Cataloguing in Publication TOXIC : cyanobacterial monitoring and cyanotoxin analysis / Jussi Meriluoto and Geoffrey A. Codd (editors). – Åbo : Åbo Akademi University Press, 2005. – (Acta Academiae Aboensis. Ser. B, Mathematica et physica, ISSN 0001-5105; vol. 65, no. 1) ISBN 951-765-259-3 ISSN 0001-5105 ISBN 951-765-259-3 Painotalo Gillot Oy Åbo 2005 i Preface This manual serves as a practical guide for the monitoring of harmful cyanobacteria (blue-green algae) and their toxins. It is one of the deliverables of the research project "TOXIC - Barriers against Cyanotoxins in Drinking Water". The TOXIC project was funded by the European Commission under the Fifth Framework Programme (contract number EVK1-CT-2002-00107) in 2002-2005. The TOXIC project involved ten European research groups in nine countries and comprised four programmes focussing on Raw Water Quality, Analysis, Treatment and Exploitation. This manual presents the core methods which have been developed, standardised and used within the Raw Water Quality and Analysis programmes. The research on raw water quality was coordinated by Prof. Geoffrey A. Codd at the University of Dundee, UK, while the analytical work was coordinated by Dr. Jussi Meriluoto at Åbo Akademi University, Turku, Finland. The book addresses an increasing worldwide need for standard protocols for cyanotoxin monitoring in lakes, reservoirs and treated waters. Toxic cyanobacteria are encountered around the world, and problems related to safe drinking water production are common. The presence of cyanobacterial toxins in drinking and bathing waters has been recognised as a human health hazard by the World Health Organization, and a provisional guideline value for the common hepatotoxin, microcystin-LR, in drinking water has been established. National legislation has been recently introduced in some European countries and elsewhere to control microcystins. Chronic exposure to low levels of cyanotoxins in drinking water is a health risk which is still only partially understood. Sample preparation protocols and analytical methods for microcystins, anatoxin-a and cylindrospermopsin have been developed and refined in the TOXIC project. These results, together with field data generated in the raw water quality subproject and effective water treatment options developed in the treatment-related workpackages of TOXIC, contribute to consumer health risk evaluation and minimisation. The coordinator of the TOXIC project was Dr. Wido Schmidt, DVGW Technologiezentrum Wasser, Branch Office Dresden. The authors of this book are indebted to Dr. Schmidt for his enthusiasm and project coordination done in a proficient and constructive manner. We would also like to thank all project partners for fruitful collaboration in the laboratory and field, and for their written contributions to this manual. This book is dedicated to the memory of Dr. Malgorzata Tarczynska. The information contained in this book does not necessarily reflect the position or opinion of the European Commission. The European Commission cannot be held liable for any use that may be made of the information contained herein. ii iii Contents Preface .....................................................................................................................i Contributors ...........................................................................................................v Abbreviations .......................................................................................................vii 1 Basic guide to detection and monitoring of potentially toxic cyanobacteria ... 1 Tomas Kull and Tore Lindholm, AAU 2 Practical guidance for environmental sampling of cyanobacteria and .......... 11 cyanobacterial toxins James S. Metcalf, Marianne Reilly and Geoffrey A. Codd, UDU 3 Microcystins and nodularins ........................................................................... 15 Lisa Spoof, AAU 4 Anatoxin-a ........................................................................................................ 41 James S. Metcalf and Geoffrey A. Codd, UDU 5 Cylindrospermopsin ......................................................................................... 43 James S. Metcalf and Geoffrey A. Codd, UDU 6 Analysis of other cyanobacterial toxins ........................................................... 47 James S. Metcalf and Geoffrey A. Codd, UDU 7 Selection of analytical methodology ................................................................ 49 Jussi Meriluoto, AAU 8 Applications and significance of results .......................................................... 51 Geoffrey A. Codd and James S. Metcalf, UDU 9 List of standard operating procedures (SOPs) ............................................... 59 SOPs, 22 contributions from AAU, UDU, ULO, TZW ......................................... 61 Annex 1. Validation of microcystin-related SOPs through intercalibration .. 147 exercises Olli Sjövall, Lisa Spoof and Jussi Meriluoto, AAU Annex 2. Best practice guidance for management of cyanotoxins in water .... 149 supplies Tom Hall, WRc iv v Contributors Peter Backlund, Department of Biochemistry and Pharmacy, Åbo Akademi University, Turku, Finland (current affiliation: Turku Regional Institute of Occupational Health, Turku, Finland) Geoffrey A. Codd, Division of Environmental and Applied Biology, School of Life Sciences, University of Dundee, Dundee, UK Tom Hall, WRc plc, Swindon, UK Katarzyna Izydorczyk, Department of Applied Ecology, University of Lodz, Lodz, Poland Tomas Kull, Department of Biology, Åbo Akademi University, Turku, Finland Tore Lindholm, Department of Biology, Åbo Akademi University, Turku, Finland Jussi Meriluoto, Department of Biochemistry and Pharmacy, Åbo Akademi University, Turku, Finland James S. Metcalf, Division of Environmental and Applied Biology, School of Life Sciences, University of Dundee, Dundee, UK Marianne Reilly, Division of Environmental and Applied Biology, School of Life Sciences, University of Dundee, Dundee, UK Wido Schmidt, DVGW Technologiezentrum Wasser, Branch Office Dresden, Germany Olli Sjövall, Department of Biochemistry and Pharmacy, Åbo Akademi University, Turku, Finland Lisa Spoof, Department of Biochemistry and Pharmacy, Åbo Akademi University, Turku, Finland Malgorzata Tarczynska, Department of Applied Ecology, University of Lodz, Lodz, Poland Deceased vi vii Abbreviations AAU .......................... Åbo Akademi University ACN........................... Acetonitrile Adda........................... 3-Amino-9-methoxy-2,6,8-trimethyl-10-phenyldeca- 4(E),6(E)-dienoic acid Antx-a ........................ Anatoxin-a CE .............................. Capillary electrophoresis Chl-a........................... Chlorophyll-a CID............................. Collision-induced dissociation CZE............................ Capillary zone electrophoresis Cyn ............................ Cylindrospermopsin DAD........................... Diode-array detector Dm (dm)..................... Demethyl DW............................. Dry weight ELISA ........................ Enzyme-linked immunosorbent assay ESI ............................. Electrospray ionisation FAB............................ Fast atom bombardment FID............................. Flame ionisation detector FISH .......................... Fluorescent in situ hybridisation Glu ............................. Glutamic acid GV ............................. Guideline value HPLC ......................... High-performance liquid chromatography i.p. ............................. Intraperitoneal IT................................ Ion-trap LC-MS ....................... Liquid chromatography-mass spectrometry LC-MS-MS................. Liquid chromatography-tandem mass spectrometry LD50 ........................... Lethal dose that kills 50% of the test organisms LLE............................ Liquid-liquid extraction LOAEL....................... Lowest observed adverse effect level LPS ............................ Lipopolysaccharide MALDI-TOF.............. Matrix-assisted laser desorption/ionisation-time-of-flight MC, MCYST.............. Microcystin Mdha .......................... N-methyldehydroalanine Mdhb.......................... 2-(Methylamino)-2(Z)-dehydrobutyric acid MEKC........................ Micellar electrokinetic chromatography MeOH ........................ Methanol MIP ............................ Molecular imprinted polymer MMPB........................ 3-Methoxy-2-methyl-4-phenylbutyric acid MRM.......................... Multiple reaction monitoring MS.............................. Mass spectrometry m/z.............................. Mass-to-charge ratio N-tot........................... Total nitrogen P-tot............................ Total phosphorus PDA .......................... Photodiode-array PPIA........................... Protein phosphatase inhibition assay Q ................................ Single-quadrupole QQQ........................... Triple-quadrupole RP .............................. Reversed-phase viii S/N ............................. Signal-to-noise SELDI-TOF...............
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