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Assessing potential health risks from microcystin toxins in blue-green algae dietary supplements.

The presence of blue-green algae (BGA) toxins in surface waters used for drinking water sources and recreation is receiving increasing attention around the world as a public health concern. However, potential risks from exposure to these toxins in contaminated health food products that contain BGA have been largely ignored. BGA products are commonly consumed in the United States, Canada, and Europe for their putative beneficial effects, including increased energy and elevated mood. Many of these products contain Aphanizomenon flos-aquae, a BGA that is harvested from Upper Klamath Lake (UKL) in southern Oregon, where the growth of a toxic BGA, Microcystis aeruginosa, is a regular occurrence. M. aeruginosa produces compounds called microcystins, which are potent hepatotoxins and probable tumor promoters. Because M. aeruginosa coexists with A. flos-aquae, it can be collected inadvertently during the harvesting process, resulting in microcystin contamination of BGA products. In fall 1996, the Oregon Health Division learned that UKL was experiencing an extensive M. aeruginosa bloom, and an advisory was issued recommending against water contact. The advisory prompted calls from consumers of BGA products, who expressed concern about possible contamination of these products with microcystins. In response, the Oregon Health Division and the Oregon Department of Agriculture established a regulatory limit of 1 microg/g for microcystins in BGA-containing products and tested BGA products for the presence of microcystins. Microcystins were detected in 85 of 87 samples tested, with 63 samples (72%) containing concentrations > 1 microg/g. HPLC and ELISA tentatively identified microcystin-LR, the most toxic microcystin variant, as the predominant congener.

Bacterial Toxins↗

Interaction of microcystin-LR with SuperChar: water decontamination and therapy.

Activated charcoal (SuperChar) has been recommended for therapeutic use against poisoning by several toxic agents, but it has not been tested against microcystin-LR toxicosis. Microcystin-LR, a cyclic heptapeptide isolated from fresh water blue-green algae, has been shown to be a potent hepatotoxin in animals and in man. Studies were performed to determine the degree of in vitro adsorption of microcystin-LR to SuperChar and to assess the efficacy of SuperChar as a therapeutic agent against microcystin-LR in vivo. Scatchard analysis of the in vitro data showed that microcystin-LR bound to SuperChar with a maximum binding capacity of 0.692 mM toxin/g SuperChar with a dissociation constant of 0.016 mM. The adsorption characteristics of microcystin-LR by SuperChar was applied successfully to the decontamination of water samples spiked with microcystin-LR. While an oral (po) dose of toxin mixed with SuperChar (0.31-0.36 g/kg) modulated the toxicity, an oral pretreatment with SuperChar did not prevent lethality induced by an oral or intraperitoneal (ip) dose of microcystin-LR in mice.

Animals↗

[Drinking water types, microcystins and colorectal cancer].

OBJECTIVE: To assess the relationship between microcystins (a blue-green algal toxin) in drinking water and colorectal cancer. METHODS: Eight townships were randomly selected as study sites in Haining city of Zhejiang province, China. Four hundred and eight colon and rectum carcinoma cases diagnosed from 1977 to 1996 were identified from cancer registry in the study sites. A retrospective survey on types of drinking water of all 408 cases was conducted. Population data and drinking water sources data were provided by local household registration and local health institution, respectively. Water samples from different sources (well, tap-water, river and pond) were collected and microcystins concentrations were measured by an indirect competitive ELISA method. RESULTS: The incidence rates of colorectal cancer were significantly higher among people drinking river or pond water than those who drank well water or tap-water in both males and females. And comparing with other sources to well water, the relative risks (RR) were much higher for people using river water (7.94) and pond water (7.70). The positive detection rates (> 50 pg/ml) of microcystin in well, tap-water, river and pond water were 0.00%, 0.00%, 36.23% and 17.14% respectively. The highest concentrations of microcystins were 1 083.43 pg/ml (river) and 1 937.94 pg/ml (pond) in the positive samples. Microcystins concentrations in river and pond were significantly higher than the concentrations in well and tap water (P < 0.01). The Spearman rank correlation analysis showed that in the study sites, the microcystins concentrations of river and pond water were positively correlated with the incidence of colorectal cancer (r = 0.881, P < 0.01). CONCLUSION: Drinking surface water (river or pond) is one of the risk factors for colorectal cancer. Microcystins may be associated with incidence of colorectal cancer. It is suggested that further study should be carried out to clarify the relationship between colorectal cancer and microcystin in drinking water.

Bacterial Toxins↗

[Microcystin contamination of fish on Poyang Lake in Jiangxi province].

In order to investigate the level of microcystin contamination of fish and its dynamic change in fish body, samples of freshwater as well as fish were collected from Poyang lake in Jiangxi province in July and October in 2000 respectively. Microcystin were tested for all samples via ELISA method. The results showed that: all freshwater samples collected in July and October contained blue-green alga and it has become a dominant alga. Microsystin content was higher in freshwater collected in October than that of in July. The average content level of microcystin in freshwater of river bank and Yongxiu river was 888.8 ng/ml, being ten times higher than that of water of Bang lake 1, Bang lake 2 and Big lake pool (83.6 pg/ml). The highest level of microcystin was 1036.9 pg/ml in freshwater of Yongxiu river. Microcystin could be detected in all samples of fish muscle and fish liver. The level of microcystin in muscle of fish collected in October was 1.2 to 2.0 times higher of that collected in July, and microcystin in liver of fish collected in October was 2.0 to 20 times higher of that collected in July. The average level of microcystin in fish liver in river bank and Yongxiu river was 27.2 ng/g, being significantly higher than that of in Bang lake1, Bang lake 2 and Big lake pool (2.8 ng/g).

Animals↗

Effect of microcystin on growth of single species and on mixed natural populations of heterotrophic nanoflagellates.

Natural populations of heterotrophic nanoflagellates were reduced in numbers and in situ growth rates during a toxic Microcystis bloom in a eutrophic lake. Microcystin was found in the particulate phase (algae) and in the dissolved phase (water) with maximum concentrations of 414 micrograms g freeze-dried algal material-1 and 141 micrograms l-1 lake water, respectively. An average reduction in growth rates of 49% was found when comparing the growth estimates before and after the toxic bloom had peaked. Similar reductions were found in laboratory experiments when growing mixed flagellate populations from two different systems with microcystin, but without predators. Concentrations of 10 micrograms toxin 1-1 reduced the growth rates by 36-41% (significantly different from the controls) and 1 microgram toxin 1-1 reduced the growth rates by 24-28% (not significant). Thus, natural populations of heterotrophic nanoflagellates seem very sensitive to microcystin. The growth characteristic of two cultured species was also tested in the presence of microcystin. Both the cultured species, Heteromita globosa and Spumella sp., grew well at 1-10 micrograms microcystin 1-1 and Spumella sp. was able to grow at 50 micrograms microcystin 1-1. However, the growth curves indicate that the decline in numbers during the stationary phase occurred faster in the presence of microcystin. The ecological consequences of a highly sensitive protozoan community may be that larger zooplankton species (copepods and cladocerans) are affected by reduced availability of food.

Animals↗

Low-energy collisionally activated decomposition and structural characterization of cyclic heptapeptide microcystins by electrospray ionization mass spectrometry.

Characteristics of electrospray ionization mass spectrometry/collision-induced dissociation (ESIMS/CID) mass spectra of microcystins, cyanobacterial cyclic heptapeptide hepatoxins, were examined. The collision conditions showed remarkable effects on the quality of the CID mass spectra, which were divided into three patterns according to the number of Arg residues. A characteristic cleavage reaction and neutral losses of MeOH, NH3 and guanidine group(s) from the (2S,3S,8S,9S)-3-amino-9-methoxy-2,6,8-trimethyl-10-phenyldeca-4 E,6E-dienoic acid (Adda) and Arg residues were observed in the ESI and ESIMS/CID mass spectra, suggesting the most probable protonation sites in [M + H]+ and [M + 2H]2+ ions of microcystins. Microcystins with no Arg residue showed only [M + H]+ ions with a proton reacting at the methoxyl group in the Adda residue, and the ESIMS/CID/MS data revealed their structures unambiguously. The protonation site in [M + H]+ ions of microcystins with Arg residue(s) was the guanidine group. The [M + 2H]2+ ions of microcystins possessing one Arg residue had one proton on the Arg residue and probably another proton on the Adda residue, while the [M + 2H]2+ ions of microcystins having two Arg residues showed protonation at both Arg residues and the ESIMS/CID/MS data assigned their sequences. Structures of microcystins possessing one Arg residue can be assigned by ESIMS/CID/MS of [M + H]+ ions combined with those of [M + 2H]2+ ions.

Anabaena↗

Recognition of hepatotoxic homologues of Microcystin using a combination of selective adsorption media.

Microcystin, a hepatotoxin produced by cyanobacteria, was chromatographically discriminated with newly prepared selective adsorption media. Since the name Microcystin refers to up to 70 different homologues, we have developed a novel technique to prepare selective recognition sites, based on the "Fragment Imprinting Effect". The homologues of Microcystin can be individually discriminated from each other by antigen-antibody assay as well as by C18-based HPLC. This is mainly due to differences in the three dimensional structure of Microcystin homologues; however, differences in toxicity are relatively small among the homologues. Therefore we have to measure the total amount of Microcystin homologues present to determine contamination by Microcystins. We have demonstrated the first group recognition for Microcystin homologues through combination of two different selective adsorption media prepared by a fragment imprinting technique.

Bacterial Toxins↗

Genotoxicity of cyanobacterial extracts containing microcystins from Polish water reservoirs as determined by SOS chromotest and comet assay.

Toxicity of cyanobacterial blooms, an increasing problem around the world, is connected to the increase in bloom samples containing microcystins, caused by excessive eutrophication of drinking- and recreational water reservoirs. Microcystins are the most common group of cyanobacterial hepatotoxins. In Poland they are produced mainly by the Microcystis genus. The toxicity of microcystins has been well documented, but investigation into their genotoxicity has been insufficient relative to the study of their overall toxicity. Therefore, the aim of this study was the estimation and comparison of the genotoxicity of cyanobacterial extracts with microcystins (CEMs) using the SOS chromotest (bacterial test) with Escherichia coli PQ37 and the comet assay with human lymphocytes. Cyanobacterial bloom samples were collected in the summer months from two Polish water reservoirs, one at Sulejów and one at Jeziorsko. The SOS chromotest, which used prokaryotic cells (without metabolic activation), and the comet assay, which used eukaryotic cells, both indicated the potential genotoxic effect of CEMs. Cyanobacterial extracts caused DNA damage in human lymphocytes in vitro. The maximum level of DNA damage was observed after 12 h incubation with CEMs. The bacterial test indicated a dependence of the degree of CEM genotoxicity, the composition, and the concentration of microcystins in each bloom sample examined with the time of exposure. Differences between the genotoxicity of cyanobacterial extract and the standard microcystin-LR were noticeable. This was probably caused by the interaction of different microcystin variants. The results showed that CEMs from Polish water reservoirs were genotoxic, which was reflected by the stimulation of the SOS repair system in bacterial cells (SOS chromotest) and by the damage induced in DNA in human lymphocytes (comet assay).

Comet Assay↗

Physiological and biochemical analyses of microcystin-RR toxicity to the cyanobacterium Synechococcus elongatus.

Freshwater Microcystis may form dense blooms in eutrophic lakes. It is known to produce a family of related cyclic hepatopeptides (microcystins, MC) that constitute a threat to aquatic ecosystems. Most toxicological studies of microcystins have focused on aquatic animals and plants, with few examining the possible effects of microcystins on phytoplankton. In this study we chose the unicellular Synechococcus elongatus (one of the most studied and geographically most widely distributed cyanobacteria in the picoplankton) as the test material and investigated the biological parameters: growth, pigment (chlorophyll-a, phycocyanin), photosynthetic activity, nitrate reductase activity, and protein and carbohydrate content. The results revealed that microcystin-RR concentrations above 100 microg x L(-1) significantly inhibited the growth of Synechococcus elongatus. In addition, a change in color of the toxin-treated algae (chlorosis) was observed in the experiments. Furthermore, MC-RR markedly inhibited the synthesis of the pigments chlorophyll-a and phycocyanin. A drastic reduction in photochemical efficiency of PSII (F(v)/F(m)) was found after a 96-h incubation. Changes in protein and carbohydrate concentrations and in nitrate reductase activity also were observed during the exposure period. This study aimed to evaluate the mechanisms of microcystin toxicity on a cyanobacterium, according to the physiological and biochemical responses of Synechococcus elongatus to different doses of microcystin-RR. The ecological role of microcystins as an allelopathic substance also is discussed in the article.

Chlorophyll↗

Chronic toxicity and responses of several important enzymes in Daphnia magna on exposure to sublethal microcystin-LR.

In the current study, the toxicological mechanisms of microcystin-LR and its disadvantageous effects on Daphnia magna were examined. Survival rate, number of newborn, activity of several important enzymes [glutathione S-transferase (GST), lactate dehydrogenase (LDH), phosphatases, and glutathione], accumulated microcystins, and ultrastructural changes in different organs of Daphnia were monitored over the course of 21-day chronic tests. The results indicated that low concentrations of dissolved microcystin had no harmful effect on Daphnia. On the contrary, stimulatory effects were detected. In the presence of toxin at high dosage and for long-term exposure, GST and glutathione levels decreased significantly. The decreased enzyme activity in the antioxidant system probably was caused by detoxification reactions with toxins. And these processes of detoxification at the beginning of chronic tests may enable phosphatases in Daphnia magna to withstand inhibition by the toxins. At the same time, we also found that the LDH activity in test animals increased with exposure to microcystin-LR, indicating that adverse effects occurred in Daphnia. With microcystin given at a higher dosage or for a longer exposure, the effect on Daphnia magna was fatal. In the meantime, microcystin began to accumulate in Daphnia magna, and phosphatase activity started to be inhibited. From the ultrastructure results of cells in D. magna, we obtained new information: the alimentary canal may be the target organ affected by exposure of microcystins to D. magna. The results of the current study also suggested that the oxidative damage and PPI (protein phosphatase inhibition) mechanisms of vertebrates also are adapted to Daphnia.

Adaptation, Physiological↗

Effects of microcystins on and toxin degradation by Poterioochromonas sp.

A Chrysophyceae species, Poterioochromonas sp., was isolated from Microcystis cultures. This species can efficiently prey on Microcystis and can grow faster phagotrophically than autotrophically. The growth of Poterioochromonas sp. was stimulated in the presence of microcystin-LR and microcystin-RR (in concentrations ranging from 0.1 to 4 mg/L). The growth rate of Poterioochromonas was 4-5 times higher than the control, indicating the toxins serve as growth stimuli for this organism. A subculture of toxin-treated cells, however, showed low cellular viability, suggesting that growth enhancement by microcystins was not a normal process. The antioxidant enzymatic activity of Poterioochromonas sp. was screened for toxicology analysis. Glutathione, malondialdehyde, and superoxide dismutase (SOD) content was up-regulated within 8 h of exposure to microcystin-LR (500 microg/L). A high level of SOD activity during exposure to the toxin indicated that SOD was involved in decreasing oxidative stress caused by microcystin-LR. Simultaneously with growth, Poterioochromonas was able to degrade microcystin-LR even, at a toxin concentration of 4 mg/L. This putative degradation mechanism in Poterioochromonas is explored further and discussed in this article. Our findings may shed light on understanding the role of Poterioochromonas in the aquatic ecosystem, in particular, as a grazer of toxic cyanobacteria and a biodegrader for microcystins.

Biodegradation, Environmental↗

Sublethal exposure from microcystins to renal insufficiency patients in Rio de Janeiro, Brazil.

In November 2001, a cyanobacterial bloom dominated by Microcystis and Anabaena occurred in the Funil Reservoir and the Guandu River, both of which supply drinking water to Rio de Janeiro, Brazil. Using ELISA, microcystins were detected at a concentration of 0.4 microg/L in the drinking water, whereas a concentration of 0.32 microg/L was detected in activated carbon column-treated water for use at the renal dialysis center of Clementino Fraga Filho Hospital (HUCFF) at the Federal University of Rio de Janeiro. A total of 44 hemodialysis patients who received care at this center were believed to be exposed. Initial ELISA analyses confirmed the presence of serum microcystin concentrations > or = 0.16 ng/mL in 90% of serum samples collected from these patients. Twelve patients were selected for continued monitoring over the following 2-month period. Serum microcystin concentrations ranged from < 0.16 to 0.96 ng/mL during the 57 days after documented exposure. ELISA-positive samples were found throughout the monitoring period, with the highest values detected 1 month after initial exposure. ESI LC/MS analyses indicated microcystins in the serum; however, MS/MS fragmentation patterns typical of microcystins were not identified. LC/MS analyses of MMPB for control serum spiked with MCYST-LR. and patient sera revealed a peak at retention time of 8.4 min and a mass of 207 m/z. These peaks are equivalent to the peak observed in the MMPB standard analysis. Taken together ELISA, LC/MS, and MMPB results indicate that these renal dialysis patients were exposed to microcystins. This documents another incident of human microcystin exposure during hemodialysis treatment.

Bacterial Toxins↗

Evaluating genotoxicity associated with microcystin-LR and its risk to source water safety in Meiliang Bay, Taihu Lake.

In recent years cyanobacteria blooms have become a severe problem in Taihu Lake, a large shallow eutrophic lake in China. Microcystins produced by certain genera of cyanobacteria can affect public health in this area because of their acute and chronic toxic effects. In this study, samples of cyanobacteria were collected and extracted by two solvent systems. The extracts were tested with three short-term genotoxicity assays, the ara test, the Ames test, and the SOS/umu test. In addition, temporal variation in the concentrations of microcystin-LR in the water samples was determined and monitored by an ELISA assay. Then the concentration of microcystin-LR in the drinking water was estimated. The risk of microcystin-LR exposure by drinking water was assessed according to tolerable daily intake (TDI). The three genotoxicity assays showed negative results regardless of the solvent system used, and there were clear inconsistencies in the spatiotemporal profiles of genotoxic potential and microcystin concentrations in Taihu Lake. Risk assessment showed that the drinking water from Taihu Lake was not safe from the end of July to the beginning of November because of a high concentration of microcystin-LR. Our study indicated the drinking water from Taihu Lake posed a risk because of the microcystin-LR, although it was neither genotoxic nor associated with genotoxicity of the lake water.

Environmental Monitoring↗

Suppression by carotenoids of microcystin-induced morphological changes in mouse hepatocytes.

Microcystin-LR is a liver tumor promoter in the okadaic acid class, a group of potent inhibitors of protein phosphatases 1 and 2A. Because of inhibition of protein phosphatases, microcystin-LR induces hyperphosphorylation of cellular proteins, including cytoskeletal proteins--cytokeratins 8 and 18--and causes morphological changes in mouse hepatocytes in primary culture. We studied the effects of carotenoids to antagonize microcystin-LR-induced morphological changes in hepatocytes. beta-carotene (100 nM to 100 microns) suppressed the morphological changes induced by 100 nM microcystin-LR in a dose-dependent manner. Other carotenoids tested exerted similar suppressive effects, although retinoids, such as all-trans retinol, all-trans retinoic acid, and 9-cis retinoic acid, were only weakly suppressive. The relative potency of the suppression correlated significantly with the number of conjugated double bonds in the trans configuration. beta-carotene strongly suppressed the hyperphosphorylation of cellular proteins induced by microcystin-LR without significant changes in the basal phosphorylation level. Other antioxidants, such as alpha-tocopherol, did not protect the cells against microcystin-LR. Taken together, the antagonistic effects of carotenoids against microcystin-LR are difficult to explain by their antioxidant or provitamin A activities. Suppression of the hyperphosphorylation of cellular proteins may be a novel mechanism by which carotenoids inhibit tumor promotion.

Animals↗

Molecular identification and evolution of the cyclic peptide hepatotoxins, microcystin and nodularin, synthetase genes in three orders of cyanobacteria.

The cyanobacterial hepatotoxins, microcystin and nodularin, are produced by a wide range of cyanobacteria. Microcystin production has been reported in the four cyanobacterial orders: Oscillatoriales, Chroococcales, Stigonematales, and Nostocales. The production of nodularin is a distinct characteristic of the Nostocales genus Nodularia. A single rapid method is needed to reliably detect cyanobacteria that are potentially capable of producing these hepatotoxins. To this end, a PCR was designed to detect all potential microcystin and nodularin-producing cyanobacteria from laboratory cultures as well as in harmful algal blooms. The aminotransferase (AMT) domain, which is located on the modules mcyE and ndaF of the microcystin and nodularin synthetase enzyme complexes, respectively, was chosen as the target sequence because of its essential function in the synthesis of all microcystins as well as nodularins. Using the described PCR, it was possible to amplify a 472 bp PCR product from the AMT domains of all tested hepatotoxic species and bloom samples. Sequence data provided further insight into the evolution of the microcystin and nodularin synthetases through bioinformatic analyses of the AMT in microcystin and nodularin synthetases, with congruence between the evolution of 16S rRNA and the AMT domain.

Bacterial Proteins↗

Cyclosporine A inhibition of microcystin toxins.

Cyclosporine A (CyA) given i.v. at a dose of 1.25 mg/mouse blocks a subsequent i.v. lethal dose (1.7-1.8 x LD50) of microcystin-LR for 24 hr, and is about 50% protective at 48 hr. Conversely, the fraction of mice that can be rescued by CyA (0.2 mg/mouse) after a lethal dose of microcystin-LR decreases rapidly with a pharmacodynamic half-time of only about 100 sec. The prophylactic action of CyA was tested against lethal doses of four microcystins. The acute lethality of 1.7-1.8 x LD50 dose of microcystin-LR, -RR, -LY, or -LA given 1 hr after administration of 0.2 mg of CyA is 0%, 0%, 58%, or 100%, respectively. Even a 0.6 mg/mouse dose of CyA is ineffective prophylaxis against a lethal dose of microcystin-LA. The inhibitory potency of CyA on microcystin toxicity can be completely reversed by the single L-amino acid substitution of alanine for arginine in the microcystin.

Animals↗

Changes in microcystin production by Microcystis aeruginosa exposed to phytoplanktivorous and omnivorous fish.

With direct exposure to phytoplanktivorous fish (Hypophthalmichthys molitrix), increased mass-specific microcystin production occurred in three monoclonal Microcystis aeruginosa strains (NIES 44, 88 and 99). Total mass-specific microcystin content of NIES 44 exposed to H. molitrix was over 50 times higher than controls (a mean value of 16.2 microgg(-1)-dry cell in controls versus 878.6 microgg(-1)-dry cell in treatments). Up to nine times higher microcystin levels were detected in NIES 88 exposed to H. molitrix compared to controls (a mean value of 553 in controls versus 5145 microgg(-1)-dry cell in treatments). The microcystin levels of all strains were significantly different between controls and H. molitrix treatments (P < 0.01 for NIES 44 and 88; P < 0.05 for NIES 99). The microcystin response to the omnivorous Carassius gibelio langsdorfi was weaker than that of H. molitrix, though the levels in all strains exposed to the fish were higher than in controls and a significant difference in microcystin production between controls and omnivorous fish treatments occurred for NIES 44 (a mean value of 6.9 in controls versus 41.5 microgg(-1)-dry cell in treatments; P < 0.01) and NIES 88 (a mean value of 359.8 versus 480.4 microgg(-1)-dry cell; P < 0.05). Microcystis cells were observed in the both fish faeces and gut contents, and microcystin was also detected in the body tissues (from 0.6 to 2.5 microgg(-1)-dry weight) and faeces of both fish species on the final day of experiment, although 98% of fish in three strains of Microcystis cultures had lost weight (mean +/- S.E. fish growth rate with M. aeruginosa; -0.90 +/- 0.06% per day, n = 96). This study showed that several M. aeruginosa strains increased toxin production when exposed to fish, especially phytoplanktivorous species, even though fish appeared not to feed vigorously on toxic Microcystis, and supports the hypothesis that this response is a fish-induced defence mediated by physical contact associated with feeding or by chemical cues (e.g. kairomones).

Analysis of Variance↗

Oral toxicity of the microcystin-containing cyanobacterium Planktothrix rubescens in European whitefish (Coregonus lavaretus).

The microcystin-producing cyanobacterium Planktothrix is one of the most widespread genera amongst toxin producing cyanobacteria in European lakes. In particular, the metalimnic blooms of Planktothrix rubescens have been associated with growing problems in the professional freshwater fishery as a decrease in yearly yields in the important coregonids fishery often coincides with the appearance of P. rubescens. P. rubescens is a cyanobacterial species known to produce toxic compounds, e.g. microcystins. Although microcystins have been reported to affect fish health, behaviour, development and growth and have also been associated with feral fish kills, there is currently no specific information on the effects of toxic Planktothrix filaments in fish and especially coregonids. Therefore, the aim of this study was to investigate the effects of an environmentally relevant dose of P. rubescens filaments orally applied to coregonids and to discuss the findings in the context of microcystin toxicity previously reported in carp and trout. A single dose of P. rubescens culture, at a density of 80,000 cells per 120 microl, was applied to coregonids thus corresponding to 0.6-0.9 microg microcystin-LR(equiv.)/kg body weight. Behavioural changes and opercular beat rates, growth, hepatosomatic index, condition and plasma glucose were determined. Liver, kidney, gill and the gastrointestinal tract were assessed histopathologically and immunohistologically. Exposed fish showed behavioural changes, increased opercular beat rates and elevated plasma glucose levels, possibly representing a physiological stress response. Histopathological alterations in liver, gastrointestinal tract and kidney, also immunopositive for microcystin suggested causality of tissue damage and the in situ presence of microcystins. The observed combination of stress and organ damage may explain the frequently reduced weight and thus the fitness noted in coregonids subjected to regular occurrences of stratified and dispersed P. rubescens blooms, e.g. in lake Ammersee, Bavaria, Germany.

Administration, Oral↗