Ingrid Chorus

Toxic Cyanobacteria in Water: A guide to their public health consequences,monitoring and management

This book describes the present state of knowledge regarding the impact of cyanobacteria on health through the use of water. It considers aspects of risk management and details the information needed for protecting drinking water sources and recreational water bodies from the health hazards caused by cyanobacteria and their toxins. It also outlines the state of knowledge regarding the principal considerations in the design of programmes and studies for monitoring water resources and supplies and describes the approaches and procedures used. The development of this publication was guided by the recommendations of several expert meetings concerning drinking water (Geneva, December 1995; Bad Elster, June 1996) and recreational water (Bad Elster, June 1996; St Helier, May 1997). An expert meeting in Bad Elster, April 1997, critically reviewed the literature concerning the toxicity of cyanotoxins and developed the scope and content of this book. A draft manuscript was reviewed at an editorial meeting in November 1997, and a further draft was reviewed by the working group responsible for updating the Guidelines for Drinkingwater Quality in March 1998.

HEALTH RISKS CAUSED BY FRESHWATER CYANOBACTERIA IN RECREATIONAL WATERS

Toxic cyanobacteria are increasingly being perceived as a potential health hazard, particularly in waters used for recreation. A few countries are developing regulations to protect human health from these toxins, and the World Health Organization (WHO) has published both a guideline value for one cyanotoxin in drinking water and a procedural guideline for recreational waters. This article presents an overview of the currently known cyanotoxins and of documented cases of human illnesses attributed to them. It further discusses exposure pathways and approaches to risk management. In this context, the WHO guideline for recreational waters is presented, and monitoring approaches are outlined.Toxic cyanobacteria are increasingly being perceived as a potential health hazard, particularly in waters used for recreation. A few countries are developing regulations to protect human health from these toxins, and the World Health Organization (WHO) has published both a guideline value for one cyanotoxin in drinking water and a procedural guideline for recreational waters. This article presents an overview of the currently known cyanotoxins and of documented cases of human illnesses attributed to them. It further discusses exposure pathways and approaches to risk management. In this context, the WHO guideline for recreational waters is presented, and monitoring approaches are outlined.

Diversity of microcystin genes within a population of the toxic cyanobacterium Microcystis spp. in Lake Wannsee (Berlin, Germany).

In order to find out how many genotypes determine microcystin production of Microcystis spp. in field populations, single colonies (clones) were sampled from Lake Wannsee (Berlin, Germany), characterized morphologically, and subsequently analyzed by PCR for a region within the mcyB gene encoding the activation of one amino acid during microcystin biosynthesis. The different morphospecies varied considerably in the proportion of microcystin-producing genotypes. Most colonies (73%) of M. aeruginosa contained this gene whereas only 16% of the colonies assigned to M. ichthyoblabe and no colonies of M. wesenbergii gave a PCR product of the mcyB gene. Restriction fragment length polymorphism revealed seven restriction profiles showing low variability in nucleotide sequence within each restriction type (0.4-4%) and a low to high variability (1.6-38%) between restriction types. In addition, the sequences of amino acids within the mcyB gene were analyzed to compare the specificity of the amino acid activation during microcystin biosynthesis between restriction types and with the occurrence of amino acids in microcystin variants as detected by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. Most of the microcystin-producing colonies showed high similarity in the sequence of amino acids and contained microcystin-LR (LR refers to leucine and arginine in the variable positions of the heptapeptide), microcystin-RR, and microcystin-YR, as well as other variants in minor concentrations. It is concluded that the gene product found for most of the microcystin-producing colonies in the lake is rather unspecific and the diversity of microcystin variants in the lake results from activation of various amino acids during microcystin biosynthesis in the same genotypes.

The Abundance of Microcystin-Producing Genotypes Correlates Positively with Colony Size in Microcystis sp. and Determines Its Microcystin Net Production in Lake Wannsee

ABSTRACT The working hypotheses tested on a natural population of Microcystis sp. in Lake Wannsee (Berlin, Germany) were that (i) the varying abundance of microcystin-producing genotypes versus non-microcystin-producing genotypes is a key factor for microcystin net production and (ii) the occurrence of a gene for microcystin net production is related to colony morphology, particularly colony size. To test these hypotheses, samples were fractionated by colony size with a sieving procedure during the summer of 2000. Each colony size class was analyzed for cell numbers, the proportion of microcystin-producing genotypes, and microcystin concentrations. The smallest size class of Microcystis colonies (<50 μm) showed the lowest proportion of microcystin-producing genotypes, the highest proportion of non-microcystin-producing cells, and the lowest microcystin cell quotas (sum of microcystins RR, YR, LR, and WR). In contrast, the larger size classes of Microcystis colonies (>100 μm) showed the highest proportion of microcystin-producing genotypes, the lowest proportion of non-microcystin-producing cells, and the highest microcystin cell quotas. The microcystin net production rate was nearly one to one positively related to the population growth rate for the larger colony size classes (>100 μm); however, no relationship could be found for the smaller size classes. It was concluded that the variations found in microcystin net production between colony size classes are chiefly due to differences in genotype composition and that the microcystin net production in the lake is mainly influenced by the abundance of the larger (>100-μm) microcystin-producing colonies.

Cylindrospermopsin occurrence in two German lakes and preliminary assessment of toxicity and toxin production of Cylindrospermopsis raciborskii (Cyanobacteria) isolates

Cylindrospermopsis raciborskii, a freshwater cyanobacterium of tropical origin, is not only increasingly found in (sub) tropical water bodies, but also in temperate regions. Since this species may produce potent toxins such as cylindrospermopsin (CYN) and paralytic shellfish poisons, its massive occurrence in water bodies used as drinking water sources or for recreation is of major concern. The proliferation of C. raciborskii in German water bodies has been documented for the past decade. We investigated the occurrence of CYN in field populations and isolates of C. raciborskii from two lakes, and assessed the toxicity of culture isolates using the mouse bioassay, primary rat hepatocytes and human derived cell lines. We show for the first time the occurrence of CYN in German water bodies. None of seven isolates of C. raciborskii contained CYN, however, all isolates were toxic to primary rat hepatocytes, human hepatoblastoma (HEP-G2) and human colon adenocarcinoma (CACO-2) cells. Methanolic extracts were more toxic than aqueous extracts. Three isolates tested in the mouse bioassay were toxic at a concentration of 800 mg kg(-1) showing liver and spleen damage and inflammation of the intestine. These results give strong evidence that the German isolates of C. raciborskii contain currently not identified or unknown toxins.

Distribution of microcystin-producing and non-microcystin-producing Microcystis sp. in European freshwater bodies: detection of microcystins and microcystin genes in individual colonies.

Microcystis is a well-known cyanobacterial genus frequently producing hepatotoxins named microcystins. Toxin production is encoded by microcystin genes (mcy). This study aims (i) to relate the mcy occurrence in individual colonies to the presence of microcystin, (ii) to assess whether morphological characteristics (morphospecies) are related to the occurrence of mcy genes, and (iii) to test whether there are geographical variations in morphospecies specificity and abundance of mcy genes. Individual colonies of nine different European countries were analysed by (1) morphological characteristics, (2) PCR to amplify a gene region within mcyA and mcyB indicative for microcystin biosynthesis, (3) matrix-assisted laser desorption/ionisation time-of-flight mass spectrometry (MALDI-TOF MS) to detect microcystins. Almost one hundred percent of the colonies predicted to produce microcystins by PCR analysis were found to contain microcystins. A high similarity in microcystin variants in the different colonies selected from lakes across Europe was demonstrated. The different morphospecies varied in the frequency with which they contained mcy genes. Most colonies (>75%) of M. aeruginosa and M. botrys contained the mcy genes, whereas < or = 20% of the colonies identified as M. ichthyoblabe and M. viridis gave a PCR product of the mcy genes. No colonies of M. wesenbergii gave a PCR product of either mcy gene. In addition, a positive relationship was found between the size of the colony and the frequency of those containing the mcy genes. It is concluded that the analysis of morphospecies is indicative for microcystin production, although the quantitative analysis of microcystin concentrations in water remains indispensable for hazard control.

Microcystins (hepatotoxic heptapeptides) in german fresh water bodies

In 1995 and 1996 a total of 55 German water bodies with different trophic states were investigated for the presence of potentially microcystin‐producing cyanobacteria. The seston biomass of over 500 samples was analyzed by HPLC to assess microcystin content. The highest microcystin content per dry weight was found when Planktothrix rubescens was dominant, followed by Planktothrix agardhii and Microcystis spp. The microcystin to chlorophyll‐a ratio mostly varied between 0.1 and 0.5, with maxima of 1–2. In over 70% of the samples from pelagic water, particulate microcystin concentrations were below 10 μg L−1. By contrast, spatial and temporal concentrations varied by 4 orders of magnitude (2–25,000 μg L−1) at bathing sites in 1997 during a mass development of Microcystis spp. The seasonal peak of development of Microcystis spp. occurred from June to September and of Planktothrix agardhii from September to November. Planktothrix rubescens, however, was almost perennial. The seasonal levels of these cyanobacteria were clearly reflected by microcystin concentrations in the water bodies. ©1999 John Wiley & Sons, Inc. Environ Toxicol 14: 13–22, 1999

Equilibrium/steady-state concept in phytoplankton ecology

This paper summarises the outcomes of the 13th Workshop of the International Association of Phytoplankton Taxonomy and Ecology (IAP). The authors mostly addressed their contributions on the following topics: the effect of trophic state on the attainment of a steady-state; the establishment of equilibria in deep and shallow lakes; the role of spatial heterogeneity, disturbance, and stress in the establishment of equilibrium assemblages; the mechanisms leading to the steady state; the frequency and longevity of equilibrium phases, and the role of morphological and physiological plasticity of phytoplankton in maintaining the (apparently) same populations under different environmental conditions. The composition of steady-state assemblages is compared to that of phytoplankton functional groups (coda). Those functional associations recognised as steady-state assemblages appear to be strongly K-determined in many instances.

Anatoxin‐a and neurotoxic cyanobacteria in German lakes and reservoirs

During a screening program conducted from 1995 to 1997 in Germany, 80 water bodies were analyzed, some regularly, some sporadically, for content of the neurotoxin anatoxin‐a. Analysis for the majority of the samples was differentiated between anatoxin‐a contained in the cells and that dissolved in the water. A total of 554 extracts (from 393 samples, many with analysis of seston as well as filtrate for differentiation of intra‐ and extracellular toxin content) were extracted with CH2Cl2 and derivatized with pentafluorobenzylbromide for GC‐ECD analysis with a detection limit of 2.5×10−4 μg L1. Only 22% of all analyzed samples contained anatoxin‐a, and the highest concentration found was 13.1 μg L−1 (the sum of the toxin in the cells and in the water). If anatoxin‐a was detected, it was usually found in the seston fraction, but only in some of the corresponding filtrates (with one exception, where it was detected only in the filtrate). In these cases, concentrations were often even higher than in the seston. The earliest occurrence of anatoxin‐a during the year was in May and the latest in November. Usually Anabaena and/or Aphanizomenon were present when anatoxin‐a was found. ©1999 John Wiley & Sons, Inc. Environ Toxicol 14: 117–125, 1999

Seasonal dynamics of cylindrospermopsin and cyanobacteria in two German lakes

Cylindrospermopsin (CYN) is a potent hepatotoxin produced by different cyanobacteria of the order Nostocales. Questions of major concern are: which species produce CYN, which are the seasonal patterns of CYN dynamics and how are they regulated? Therefore, we studied for the first time the seasonal dynamics of particulate and dissolved CYN concentrations, cyanobacterial abundance and environmental factors in two German lakes over 2 years. Total CYN reached maximum concentrations of 0.34 and 1.80 microg L(-1) in Melangsee and Langer See, respectively. In both lakes, maxima of the dissolved CYN fraction occurred later in the season than those of the particulate fraction, and it reached higher concentrations. This indicates that CYN is poorly decomposed and accumulates in the water. The cyanobacterial community in both lakes included several potentially CYN-producing species that did not correlate with CYN concentrations. Significant correlations between the particulate CYN concentrations and species biovolume were only found for Aphanizomenon gracile (r(s)=0.803) in Langer See indicating that this species is a CYN producer. Different correlations of CYN with abiotic factors in the two lakes indicate the presence of further undetected CYN producers as well as different regulation mechanisms of their dynamics and the variability of CYN.