HARMFUL CYANOBACTERIA

AQUATIC ECOLOGY SERIES

Volume 3

The titles published in this series are listed at the end of this volume.
HARMFUL CYANOBACTERIA

Edited by

JEF HUISMAN
Institute for Biodiversity and Ecosystem Dynamics, University of Amsterdam,
The Netherlands

and

HANS C.P. MATTHIJS

Institute for Biodiversity and Ecosystem Dynamics, University of Amsterdam,
The Netherlands

and

PETRA M. VISSER
Institute for Biodiversity and Ecosystem Dynamics, University of Amsterdam,
The Netherlands
A C.I.P. Catalogue record for this book is available from the Library of Congress.

ISBN 1-4020-3009-6 (HB)

ISBN 1-4020-3022-3 (e-book)

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Cover photo:
Dense bloom of the cyanobacterium /Microcystis/, Wann See, Germany
(photo taken by Gertrud Schlag).

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To Professor Luuc R. Mur
For his leading contributions
to European research on harmful cyanobacteria
For his guidance of a new generation of aquatic scientists
 AQUATIC ECOLOGY SERIES

Aquatic ecology is an extraordinarily broad and diverse discipline. Aquatic

ecology is the study of the functional relationships and productivity of organisms and
communities of waters as regulated by their physical, chemical, and biotic environment.
The marine environment extends broadly from the complex land-water coastal
environments to the open ocean. Limnology encompasses all inland aquatic
environments, including streams, rivers, lakes, reservoirs, and wetlands. Research has
accelerated in certain areas and been less active in others. Reassessments and syntheses
are stimulating to the discipline as a whole, as well as enormously useful to students and
researchers in ecological sciences.

A series of succinct monographs and specialized evaluations in aquatic ecology

has been developed. Subjects covered are topical (e.g., lake hydrodynamics, microbial
loop in aquatic ecosystems) rather than broad and superficial.

The treatments must be comprehensive and state-of-the-art, whether the topic is at

the biochemical, mathematical, population, community, or ecosystem level. The
objectives are to advance the topics by the development of arguments, with documented
support, that generate new insights, concepts, theories to stimulate thought, ideas,
directions, controversies. The books are intended for mature as well as emerging
scientists to stimulate intellectual leadership in the topics treated.

Receipt of manuscripts approximately 18 months after an agreement is desired,

for publication within 10 months thereafter. Electronic submission is essential with
hardcopy. Format and manuscript guidance will be provided.

For further information and book proposal details please contact:

Prof. Robert G. Wetzel, Series Editor Dr. Anna Besse, Publishing Editor
Department of Environmental Sciences Aquatic and Biogeosciences
and Engineering Springer
The University of North Carolina Van Godewijckstraat 30
Chapel Hill, North Carolina 27599-7431 P. O. Box 17
USA 3300 AA Dordrecht
Email: rwetzel@unc.eduu The Netherlands
Phone: 919 + 843-4916 Email: anna.besse@springer-sbm.com
Phone: 31 (0) 651 33 86 01
 CONTENTS

PREFACE ix

CONTRIBUTORS xi

1. HARMFUL CYANOBACTERIA 1
From mass mortalities to management measures
G.A. Codd, J. Lindsay, F.M. Young, L.F. Morrison & J.S. Metcalf

2. MOLECULAR BIOLOGY OF CYANOBACTERIAL TOXINS 25

Genetic basis of microcystin production
T. Börner & E. Dittmann

3. DYNAMICS OF CYANOBACTERIAL TOXINS 41

Sources of variability in microcystin concentrations
W.E.A. Kardinaal & P.M. Visser

4. NUTRIENT LIMITATION OF FRESHWATER CYANOBACTERIA 65

Tools to monitor phosphorus limitation at the individual level
M. Dignum, H.C.P. Matthijs, R..Pel, H.J. Laanbroek & L.R.. Mur

5. NUTRIENT LIMITATION OF MARINE CYANOBACTERIA 87

Molecular ecology of nitrogen limitation in an oligotrophic sea
A.F. Post

6. THE ECOPHYSIOLOGY OF THE HARMFUL 109

CYANOBACTERIUM MICROCYSTIS
Features explaining its success and measures for its control
P.M. Visser, B.W. Ibelings, L.R. Mur & A.E. Walsby

7. POPULATION DYNAMICS OF HARMFUL CYANOBACTERIA 143

Factors affecting species composition
J. Huisman & F.D. Hulot

8. OPTICAL DETECTION OF CYANOBACTERIAL BLOOMS 177

Shipboard observation and remote sensing
H.J. Gons, H. Hakvoort, S.W.M. Peters & S.G.H. Simis

9. WATER SAFETY PLANS 201

A better approach to prevent human exposure to harmful cyanobacteria
I. Chorus

INDEX 229
 PREFACE

Innocently, the white dog looks up: ‘What ‘s all that fuss about? Why am I called
back ashore, when I ‘m having so much fun in this green soup?’ Indeed, it is a wise
decision of the owner to call this dog back k ashore without delay. The green soup the
dog is standing in is a dense surface bloom of Microcystis. The cyanobacterium
Microcystis is a cosmopolitan species of freshwater lakes. It can produce
microcystins, a toxic substance that has caused numerous illnesses and deaths of
dogs, cattle, birds, and even humans.
Are all cyanobacteria harmful? No, not at all. Cyanobacteria are phototrophic
microorganisms that convert sunlight into organic biomass by means of
photosynthesis. As such, cyanobacteria are the aquatic equivalents of grasses and
trees on land. In fact, cyanobacteria were the first organisms in the evolutionary
history of our planet to produce oxygen. They are the evolutionary ancestors of all
modern plants. Together with the eukaryotic phytoplankton, cyanobacteria form the
basis of the aquatic food web.
However, a few cyanobacterial species are capable of producing powerful toxins.
Several of these harmful species are quite widespread. They occur throughout the
world, especially in fresh and brackish waters, and may occasionally reach high
population densities, as the cover of this book illustrates. Waters dominated by such
harmful cyanobacteria are often closed for recreation, and cannot be used for the
intake of drinking water or agricultural purposes. During the past few years, much
new knowledge on harmful cyanobacteria has been acquired, owing to scientific
advances in molecular biology, genomics, and the computational sciences and also
owing to an increased recognition of the problems caused by harmful cyanobacteria.
In fact, it is fair to say that we now know a lot more about harmful cyanobacteria
than only five years ago. The aim of this book, therefore, is to provide an up-to-date
review that makes this recently acquired knowledge accessible to a broad readership.
The first chapter (chapter 1) in this book introduces the subject, and explains the
problems associated with harmful cyanobacteria. Two subsequent chapters (chapters
2 and 3) focus on the molecular biology and physiology of toxin production, with
emphasis on the genetic regulation and variability in toxin production. Next, modern
approaches are discussed that allow rapid assessment of the extent to which
nutrients, such as nitrogen and phosphorus, limit the growth of cyanobacteria
(chapters 4 and 5). The subsequent chapter (chapter 6) reviews our current
knowledge of the physiology and ecology of Microcystis, one of the most
widespread and extensively studied harmful species. This is followed by a state-of-
the-art review of the population dynamics of harmful cyanobacteria, with particular
attention to environmental factors that favour cyanobacteria over other
phytoplankton species (chapter 7). Chapter 8 introduces novel techniques in remote
sensing, which allow monitoring of the development of cyanobacterial blooms from
aircraft and satellites. The final chapter (chapter 9) discusses how improvements in
water management, based on the new Water Safety Plans introduced by the World
Health Organization, may reduce human exposure to harmful cyanobacteria.

ix
x PREFACE

The idea for this book came up during a farewell symposium, organized at the
University of Amsterdam in September 2003, on the occasion of the retirement of
Professor Luuc Mur. For more than 40 years, Luuc Mur has been active in
phytoplankton biology, first as lecturer and later as Professor in Aquatic
Environmental Biology at the University of Amsterdam. He has educated numerous
undergraduate and graduate students on water management, the marvellous beauty
of the phytoplankton, and the potential trouble of harmful cyanobacteria. Many of
these former students are still very active in aquatic research and water management
today. During the 1970s and 1980s, Luuc Mur was among the first scientists to
recognize that advances in microbiology could be made useful in aquatic ecology. In
particular, phytoplankton species such as the cyanobacteria and green algae could be
studied in the laboratory using continuous-culture techniques. This culture technique
enabled detailed investigation of the growth kinetics of these microorganisms under
controlled laboratory conditions. In the 1980s and 1990s, with the increasing
eutrophication of many European lakes, the problems caused by harmful
cyanobacteria became widely apparent. With funding provided by the fourth
framework program of the European Committee, Luuc Mur initiated two research
projects (CYANOTOX and TOPIC) to study the toxicology, molecular biology,
physiology, and ecology of harmful cyanobacteria. These two research projects
brought together a diverse group of European scientists that have stood at the basis
of much of what is currently known about harmful cyanobacteria. It is our pleasure -
on behalf of all colleagues, students and friends - to thank Luuc Mur for his leading
contributions and guidance of a new generation of aquatic scientists.
This book would not have been possible without the financial support provided
by the Earth and Life Sciences Foundation (ALW), the Netherlands Organization for
Scientific Research (NWO), and the University of Amsterdam.
Special thanks go to all chapter authors, whose work provides the backbone of
this book, and to all reviewers that have been of great help in sharpening the
chapters. We thank Bibi Krot and Pascale Thiery for helping us in organizing the
symposium, and for their invaluable assistance during the development of this book.
We thank Gertrud Schlag for providing the beautiful photograph that illustrates the
cover, and Ellen Spanjaard for the cover design. Last but not least, we are most
grateful to Professor Robert Wetzel and the staff at Springer, especially Anna Besse-
Lototskaya and Judith Terpos, for their stimulating guidance.

Jef Huisman, Hans C.P. Matthijs & Petra M. Visser (editors)

Amsterdam, August 2004.
 CONTRIBUTORS

THOMAS BÖRNER is Professor of Genetics at the Humboldt University in Berlin.

His research focuses on plant molecular biology, and in particular on the molecular
biology of harmful cyanobacteria. Address: Institut für Biologie, Humboldt
Universität, Chausseestrasse 117, D-10115 Berlin, Germany.

INGRID CHORUS is research scientist at the Federal Environmental Agency

Germany in Berlin. Her research spans many aspects of harmful cyanobacteria,
including detection methods, toxin production and water management strategies.
Address: Institute for Water, Soil and Air Hygiene, Federal Environmental Agency
Germany, Corrensplatz 1, D-14195 Berlin, Germany.

GEOFFREY A. CODD is Professor of Microbiology at the University of Dundee.

His research focuses on the analysis, biology, and toxicology of cyanobacterial
toxins, with applications in water-quality management and health. Address: Division
of Environmental and Applied Biology, School of Life Sciences, University of
Dundee, Dundee DD1 4HN, United Kingdom.

MARCO DIGNUM is postdoctoral researcher at the Netherlands Institute of

Ecology (NIOO-KNAW). He works on the mechanisms and regulation of phosphate
uptake by freshwater cyanobacteria. Address: Centre for Limnology, Netherlands
Institute of Ecology, Rijksstraatweg 6, 3631 AC Nieuwersluis, Netherlands.

ELKE DITTMANN is Junior Professor of Molecular Ecology at the Humboldt

University in Berlin. Her research addresses the biochemistry of peptide and
polyketide biosynthesis in cyanobacteria, and the functional role of cyanobacterial
secondary metabolites. Address: Institut für Biologie, Humboldt Universität,
Chausseestrasse 117, D-10115 Berlin, Germany.

HERMAN J. GONS is senior scientist at the Netherlands Institute of Ecology

(NIOO-KNAW). His research interests include the role of viruses in plankton
communities, and the application of optical teledetection to study spatial
distributions of cyanobacteria. Address: Centre for Limnology, Netherlands Institute
of Ecology, Rijksstraatweg 6, 3631 AC Nieuwersluis, Netherlands.

HANS HAKVOORT is senior advisor at the Dutch Directorate-General for Public

Works and Water Management. His work specializes on the application of
geographical information systems for monitoring of water quality. Address: Geo-
information and ICT Department, PO Box 5023, 2600 GA Delft, Netherlands.

JEF HUISMAN is Professor of Aquatic Microbiology at the University of

Amsterdam. His research focuses on the population dynamics of phytoplankton,
using mathematical models, lab experiments, and field research. Address: Aquatic
Microbiology, Institute for Biodiversity and Ecosystem Dynamics, University of
Amsterdam, Nieuwe Achtergracht 127, 1018 WS Amsterdam, Netherlands.
xi
xii CONTRIBUTORS

FLORENCE D. HULOT is assistant professor at the Ecole Normale Supérieure in

Paris. She works on species interactions and biodiversity in aquatic ecosystems,
using mathematical models and experimental research. Address: Laboratoire
d'Ecologie, Ecole Normale Supérieure, 46 rue d'Ulm, 75230 Paris cedex 05, France.

BAS W. IBELINGS is postdoctoral researcher at the Netherlands Institute of

Ecology (NIOO-KNAW). His research interests include the ecophysiology of
freshwater phytoplankton, and the evolutionary dynamics of aquatic communities.
Address: Centre for Limnology, Netherlands Institute of Ecology, Rijksstraatweg 6,
3631 AC Nieuwersluis, Netherlands.

W. EDWIN A. KARDINAAL is PhD student at the University of Amsterdam. His

research addresses the dynamics and toxin production of freshwater cyanobacteria.
Address: Aquatic Microbiology, Institute for Biodiversity and Ecosystem Dynamics,
University of Amsterdam, Nieuwe Achtergracht 127, 1018 WS Amsterdam,
Netherlands.

HENDRIKUS J. LAANBROEK is director of the Centre for Limnology of the

Netherlands Institute of Ecology (NIOO-KNAW), and Professor in Soil Biology at
the University of Nijmegen. His research focuses on the role of microorganisms in
biogeochemical cycles. Address: Centre for Limnology, Netherlands Institute of
Ecology, Rijksstraatweg 6, 3631 AC Nieuwersluis, Netherlands.

JAIME LINDSAY is PhD student at the University of Dundee. Her research

includes investigations into the characterisation and significance of cyanobacterial
lipopolysaccharides and endotoxins. Address: Division of Environmental and
Applied Biology, School of Life Sciences, University of Dundee, Dundee DD1
4HN, United Kingdom.

HANS C.P. MATTHIJS is associate professor at the University of Amsterdam. He

investigates the molecular biology and physiology of cyanobacteria, with special
emphasis on nutrient acquisition and photosynthesis. Address: Aquatic
Microbiology, Institute for Biodiversity and Ecosystem Dynamics, University of
Amsterdam, Nieuwe Achtergracht 127, 1018 WS Amsterdam, Netherlands.

JAMES S. METCALF is postdoctoral researcher at the University of Dundee. His

current research interests include the chromatographic and molecular biological
analysis of cyanobacterial toxins in a wide range of biological and clinical materials.
Address: Division of Environmental and Applied Biology, School of Life Sciences,
University of Dundee, Dundee DD1 4HN, United Kingdom.

LOUISE F. MORRISON is PhD student at the University of Dundee. Her research

includes investigations into the occurrence and environmental fate of cyanobacterial
toxins. Address: Division of Environmental and Applied Biology, School of Life
Sciences, University of Dundee, Dundee DD1 4HN, United Kingdom.
 CONTRIBUTORS xiii

LUUC R. MUR is emeritus professor at the University of Amsterdam. His research

focuses on phytoplankton biology and water management. He has initiated several
European research programmes on harmful cyanobacteria. Present address: Aquatic
Microbiology, Institute for Biodiversity and Ecosystem Dynamics, University of
Amsterdam, Nieuwe Achtergracht 127, 1018 WS Amsterdam, Netherlands.

ROEL PEL is senior scientist at the Netherlands Institute of Ecology (NIOO-

KNAW). His research focuses on the applications of stable isotopes in microbial
ecology. Address: Centre for Limnology, Netherlands Institute of Ecology,
Rijksstraatweg 6, 3631 AC Nieuwersluis, Netherlands.

STEEF W.M. PETERS is senior scientist at the Free University in Amsterdam. He is

an expert on remote sensing and geographical information systems, with a specific
interest in aquatic ecosystems. Address: Institute for Environmental Studies, Free
University, De Boelelaan 1087, 1081 HV Amsterdam, Netherlands.

ANTON F. POST is senior scientist at the Interuniversity Institute for Marine

Sciences in Eilat, Israel, and professor at the Hebrew University. He investigates the
molecular ecology and genomics of marine cyanobacteria, with a specific focus on
nutrient stress responses. Address: Interuniversity Institute for Marine Sciences,
Coral Beach, PO Box 469, Eilat 88000, Israel.

STEFAN G.H. SIMIS is PhD student at the Netherlands Institute of Ecology

(NIOO-KNAW). The focus of his work is on the remote sensing of freshwater
cyanobacteria, and the impact of viruses on cyanobacterial blooms. Address: Centre
for Limnology, Netherlands Institute of Ecology, Rijksstraatweg 6, 3631 AC
Nieuwersluis, Netherlands.

PETRA M. VISSER is assistant professor at the University of Amsterdam. She

investigates the ecophysiology and population dynamics of freshwater
cyanobacteria. Address: Aquatic Microbiology, Institute for Biodiversity and
Ecosystem Dynamics, University of Amsterdam, Nieuwe Achtergracht 127, 1018
WS Amsterdam, Netherlands.

ANTHONY E. WALSBY is Melville Wills Professor of Botany at the University of

Bristol. His research focuses on the genetics, physiology and ecology of planktonic
cyanobacteria, using a combination of molecular approaches, field research, and
mathematical modeling. Address: School of Biological Sciences, University of
Bristol, Bristol BS8 1UG, United Kingdom.

FIONA YOUNG is PhD student at the University of Dundee. Her research interests
include the localisation of cyanobacterial toxins and their potential role in
cyanobacteria. Address: Division of Environmental and Applied Biology, School of
Life Sciences, University of Dundee, Dundee DD1 4HN, United Kingdom.