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A guide to state-of-the-art molecular tools for monitoring and managing the toxigenicity of cyanobacteria
Runaway eutrophication and climate change has made the monitoring and management of toxigenic organisms in the world's bodies of water more urgent than ever. In order to influence public policy regarding the detection and quantification of those organisms, it is incumbent upon scientists to raise the awareness of policy makers concerning the increased occurrence of toxigenic cyanobacteria and the threats they pose. As molecular methods can handle many samples in short time and help identify toxigenic organisms, they are reliable, cost-effective tools available for tracking toxigenic cyanobacteria worldwide. This volume arms scientists with the tools they need to track toxigenicity in surface waters and food supplies and, hopefully, to develop new techniques for managing the spread of toxic cyanobacteria.
This handbook offers the first comprehensive treatment of molecular tools for monitoring toxigenic cyanobacteria. Growing out of the findings of the landmark European Cooperation in Science and Technology Cyanobacteria project (CYANOCOST), it provides detailed, practical coverage of the full array of available molecular tools and protocols, from water sampling, nucleic acid extraction, and downstream analysis--including PCR and qPCR based methods--to genotyping (DGGE), diagnostic microarrays, and community characterization using next-gen sequencing techniques.
* Offers an overview of the latest trends in the field, while providing a foundation for understanding and applying the tools and techniques described
* Provides detailed coverage of the full range of molecular tools currently available, with expert guidance on the analysis and interpretation of results
* Includes step-by-step guidance on standard operational procedures, including molecular tests used in environmental monitoring, with individual chapters devoted to each procedure
* Complements the published Handbook of Cyanobacterial Monitoring and Cyanotoxin Analysis from the CyanoCOST project
This handbook is an indispensable working resource for scientists, lab technicians, and water management professionals and an excellent text/reference for graduate students and supervisors who use molecular tools. It will also be of great value to environmental health and protection officials and policy makers.
List of contents
List of Contributors xix
About the Editors xxiii
About the Book xxvii
Preface xxix
Acknowledgments xxxi
1 Introduction 1
Rainer Kurmayer, Kaarina Sivonen, and Nico Salmaso
1.1 A Brief Historical Overview 1
1.2 The Genetic Basis of Toxin Production 2
1.2.1 Microcystin and Nodularin 2
1.2.2 Cylindrospermopsin 5
1.2.3 Saxitoxin 6
1.2.4 Anatoxin 8
1.3 Application of Molecular Tools 8
1.4 Laboratory Safety Issues 13
1.5 References 14
2 Sampling and Metadata 19
Rainer Kurmayer, Guntram Christiansen, Konstantinos Kormas, Wim Vyverman, Elie Verleyen, Vitor Ramos, Vitor Vasconcelos, and Nico Salmaso
2.1 Introduction 19
2.2 Handling of Samples 20
2.3 Sample Contamination 21
2.4 Sampling 21
2.4.1 Quantitative Depth-Integrated and Discrete Sampling 21
2.4.2 Qualitative Plankton Net Sampling 22
2.4.3 Surface (Scum Material) Sampling 22
2.4.4 Benthic (Terrestrial) Cyanobacteria Sampling 22
2.4.5 Food Supplement Sampling 22
2.4.6 Isolation of Single Colonies/Filaments 22
2.5 Subsampling Food Supplement Samples 23
2.6 Sampling of Nucleic Acids 23
2.7 General Conclusions 24
2.8 References 24
SOP 2.1 Sampling and Filtration (DNA) 26
Rainer Kurmayer and Konstantinos Kormas
SOP 2.1.1 Introduction 26
SOP 2.1.2 Experimental 26
SOP 2.1.3 Procedure 27
SOP 2.1.4 Notes 28
SOP 2.1.5 References 29
SOP 2.2 Sampling of Benthic Cyanobacteria 29
Wim Vyverman and Elie Verleyen
SOP 2.2.1 Introduction 29
SOP 2.2.2 Experimental 30
SOP 2.2.3 Procedure 30
SOP 2.2.4 Notes 31
SOP 2.2.5 References 31
SOP 2.3 Isolation of Single Cyanobacteria Colonies/Filaments 32
Rainer Kurmayer
SOP 2.3.1 Introduction 32
SOP 2.3.2 Experimental 32
SOP 2.3.3 Procedure 33
SOP 2.3.4 Notes 33
SOP 2.3.5 References 33
SOP 2.4 Sampling Food Supplements 34
Vitor Ramos, Cristiana Moreira, and Vitor Vasconcelos
SOP 2.4.1 Introduction 34
SOP 2.4.2 Experimental 35
SOP 2.4.3 Procedure (Fig. 8.3) 35
SOP 2.4.4 Notes 36
SOP 2.4.5 References 36
SOP 2.5 Sampling and Filtration (RNA) 37
Rainer Kurmayer and Guntram Christiansen
SOP 2.5.1 Introduction 37
SOP 2.5.2 Experimental 37
SOP 2.5.3 Procedure 38
SOP 2.5.4 Notes 38
SOP 2.5.5 References 38
SOP 2.6 Sampling of Abiotic and Biotic Data and Recording Metadata 39
Elie Verleyen, Maxime Sweetlove, Dagmar Obbels, and Wim Vyverman
SOP 2.6.1 Introduction 39
SOP 2.6.2 Experimental 39
SOP 2.6.3 Type of Metadata and Additional Biotic and Abiotic Data 40
SOP 2.6.4 Notes 41
SOP 2.6.5 References 42
3 Isolation, Purification, and Cultivation of Toxigenic Cyanobacteria 43
Sigrid Haande, Iwona Jasser, Muriel Gugger, Camilla H.C. Hagman, Annick Wilmotte, and Andreas Ballot
3.1 Introduction 43
3.2 Methodical Principles for Cyanobacterial Isolation, Purification, and Cultivation 44
3.2.1 Sampling, Identification, and Treatments Prior to the Isolation of Cyanobacteria 44
3.2.2 Traditional Techniques for the Isolation and Purification of Cyanobacteria 45
3.2.3 Culture Media Preparation 47
3.2.4 Cultivation Conditions 48
3.3 General Conclusions 49
3.4 References 49
SOP 3.1 Isolation
About the author
EDITED BY
RAINER KURMAYER, PhD, is an Associate Professor at the University of Innsbruck, Research Institute for Limnology, Mondsee, Austria.
KAARINA SIVONEN, PhD, is a Professor of Microbiology at the University of Helsinki, Finland.
ANNICK WILMOTTE, PhD, is a FRS-FNRS Research Associate at InBios - Center for Protein Engineering, University of Liège, Belgium.
NICO SALMASO, PhD, is Head of the Hydrobiology Unit of the Istituto Agrario di S. Michele All'Adige, Fondazione E. Mach (FEM), Trento, Italy.
Summary
A guide to state-of-the-art molecular tools for monitoring and managing the toxigenicity of cyanobacteria Runaway eutrophication and climate change has made the monitoring and management of toxigenic organisms in the world s bodies of water more urgent than ever.
