microcystin and anatoxin-a

Global warming and eutrophication may lead to increased incidence of harmful algal blooms and related production of cyanotoxins that can be toxic to aquatic plants. Previous studies have evaluated the phytotoxic effects of cyanotoxins on aquatic plants. However, most studies have evaluated only a limited number of plant species and cyanotoxins; there is also considerable variability between studies, which obscures general patterns and hinders understanding of the phytotoxic effects of cyanotoxins. Here, we conducted a comprehensive meta-analysis by compiling 41 published papers to estimate the phytotoxic effects of anatoxin-a, cylindrospermopsin, and microcystins in 34 species of aquatic plants, with the aim of 1) investigating the phytotoxicity of different cyanotoxins to aquatic plants; 2) determining the aquatic plant species most sensitive to the phytotoxic effects of cyanotoxins; and 3) evaluating the bioaccumulation potential of cyanotoxins in aquatic plants. Most aquatic plants were negatively affected by cyanotoxin exposure and their response was dose-dependent; however, morphological indicators and photosynthesis of certain aquatic plants were marginally stimulated under low concentrations of anatoxin-a and cylindrospermopsin. Anatoxin-a showed the greatest bioaccumulation capacity in aquatic plants compared to cylindrospermopsin and microcystin variants. Bioaccumulation factors of cyanotoxins in aquatic plants generally decreased with increasing water exposure concentrations. Our study supports the One Health goal to manage the risk of public exposure to toxic substances, and indicates that cyanotoxins warrant further investigations in aquatic plants. Environmental managers and public health authorities need to be alert to the long-term exposure and chronic toxicity of cyanotoxins, and the potential trophic transfer of cyanotoxins from aquatic plants to higher-order organisms.

Topics: Alkaloids; Bacterial Toxins; Cyanobacteria Toxins; Harmful Algal Bloom; Microcystins; Tropanes

Mass developments of cyanobacteria ("blue-green algae") in lakes and brackish waters have repeatedly led to serious concerns due to their frequent association with toxins. Among these are the widespread hepatotoxins microcystin (MC) and nodularin (NOD). Here, we give an overview about the ecostrategies of the diverse toxin-producing species and about the genes and enzymes that are involved in the biosynthesis of the cyclic peptides. We further summarize current knowledge about toxicological mechanisms of MC and NOD, including protein phosphatase inhibition, oxidative stress and their tumor-promoting capabilities. One biotransformation pathway for MC is described. Mechanisms of cyanobacterial neurotoxins (anatoxin-a, homanatoxin-a, and anatoxin-a(s)) are briefly explained. We highlight selected cases of human fatalities related to the toxins. A special focus is given to evident cases of contamination of food supplements with cyanobacterial toxins, and to the necessary precautions.

Topics: Animals; Bacterial Toxins; Carcinogens; Chemical and Drug Induced Liver Injury; Cyanobacteria; Cyanobacteria Toxins; Dietary Supplements; Ecosystem; Food Contamination; Humans; Marine Toxins; Microcystins; Oxidative Stress; Peptides, Cyclic; Phosphoprotein Phosphatases; Tropanes

In order to evaluate the recurrence of toxic cyanobacterial blooms and to determine the survival capabilities of the resistance cells through time, a sedimentary core spanning 6700 years was drilled in the eutrophic Lake Aydat. A multiproxy approach (density, magnetic susceptibility, XRF, pollen and non-pollen palynomorph analyses), was used initially to determine the sedimentation model and the land uses around the lake. Comparison with the akinete count revealed that Nostocales cyanobacteria have been present in Lake Aydat over a six thousand year period. This long-term cyanobacterial recurrence also highlights the past presence of both the anaC and mcyB genes, involved in anatoxin-a and microcystin biosynthesis, respectively, throughout the core. The first appearance of cyanobacteria seems to be linked to the natural damming of the river, while the large increase in akinete density around 1800 cal.yr BP can be correlated with the intensification of human activities (woodland clearance, crop planting, grazing, etc.) in the catchment area of the lake, and marks the beginning of a long period of eutrophication. This first investigation into the viability and germination potential of cyanobacteria over thousands of years reveals the ability of intact akinetes to undergo cell divisions even after 1800 years of sedimentation, which is 10 times longer than previously observed. This exceptional cellular resistance, coupled with the long-term eutrophic conditions of this lake, could partly explain the past and current recurrences of cyanobacterial proliferations.

Topics: Cyanobacteria; Cyanobacteria Toxins; Environmental Monitoring; Eutrophication; Geologic Sediments; Lakes; Microcystins; Recurrence; Seasons; Tropanes; Water Pollution

The aim of this study was to obtain the first data on the occurrence and distribution of potentially toxic cyanobacteria and cyanotoxins in the Russian Easternmost part of the Gulf of Finland of the Baltic Sea. Studied samples were collected from 2012 to 2017 and three independent approaches - HPLC-HRMS, PCR and light microscopy were applied for cyanotoxins analysis and detection of toxigenic cyanobacteria. Aphanizomenon flos-aquae Ralfs ex Born. et Flah., Planktothrix agardhii (Gom.) Anag. et Kom., Microcystis aeruginosa (Kütz.) Kütz. and Dolichospermum spp. dominated among cyanobacteria in collected samples. In 2012-2013 during research cruises, microcystins concentrations varied from below detection levels to low (0.01-0.6 μg L

Topics: Cyanobacteria; Cyanobacteria Toxins; DNA, Bacterial; Eutrophication; Microcystins; Oceans and Seas; Russia; Seawater; Tropanes

Cyanobacterial harmful algal blooms (CyanoHABs) are complex communities that include coexisting toxic and non-toxic strains only distinguishable by genetic methods. This study shows a water-management oriented use of next generation sequencing (NGS) to specifically pinpoint toxigenic cyanobacteria within a bloom simultaneously containing three of the most widespread cyanotoxins (the hepatotoxins microcystins, MCs; and the neurotoxins anatoxin-a, ATX, and saxitoxins, STXs). The 2013 summer bloom in Rosarito reservoir (Spain) comprised 33 cyanobacterial OTUs based on 16S rRNA metabarcoding, 7 of which accounted for as much as 96.6% of the community. Cyanotoxins and their respective biosynthesis genes were concurrently present throughout the entire bloom event including: MCs and mcyE gene; ATX and anaF gene; and STXs and sxtI gene. NGS applied to amplicons of cyanotoxin-biosynthesis genes unveiled 6 toxigenic OTUs, comprising 3 involved in MCs production (Planktothrix agardhii and 2 Microcystis spp.), 2 in ATX production (Cuspidothrix issatschenkoi and Phormidium/Tychonema spp.) and 1 in STXs production (Aphanizomenon gracile). These toxigenic taxa were also present in 16S rRNA OTUs list and their relative abundance was positively correlated with the respective toxin concentrations. Our results point at MC-producing P. agardhii and ATX-producing C. issatschenkoi as the main contributors to the moderate toxin concentrations observed, and suggest that their distribution in Southern Europe is broader than previously thought. Our findings also stress the need for monitoring low-abundance cyanobacteria (<1% relative abundance) in cyanotoxicity studies, and provide novel data on the presence of picocyanobacteria and potentially ATX-producing benthic taxa (e.g., Phormidium) in deep thermally-stratified water bodies. This study showcases a straightforward use of amplicon metagenomics of cyanotoxin biosynthesis genes in a multi-toxin bloom thus illustrating the broad applicability of NGS for water management in risk-oriented monitoring of CyanoHABs.

Topics: Aphanizomenon; Bacterial Toxins; Cyanobacteria; Cyanobacteria Toxins; Europe; Harmful Algal Bloom; High-Throughput Nucleotide Sequencing; Microcystins; Microcystis; Neurotoxins; RNA, Ribosomal, 16S; Spain; Tropanes

The eutrophication of waterways has led to a rise in cyanobacterial, harmful algal blooms (CyanoHABs) worldwide. The deterioration of water quality due to excess algal biomass in lakes has been well documented (e.g., water clarity, hypoxic conditions), but health risks associated with cyanotoxins remain largely unexplored in the absence of toxin information. This study is the first to document the presence of dissolved microcystin, anatoxin-a, cylindrospermopsin, and β-

Topics: Alkaloids; Amino Acids, Diamino; Bacterial Toxins; Cyanobacteria; Cyanobacteria Toxins; Environmental Monitoring; Harmful Algal Bloom; Lakes; Microcystins; North Carolina; Tropanes; Uracil; Water Pollutants; Water Supply

Benthic algae fuel summer food webs in many sunlit rivers, and are hotspots for primary and secondary production and biogeochemical cycling. Concerningly, riverine benthic algal assemblages can become dominated by toxic cyanobacteria, threatening water quality and public health. In the Eel River in Northern California, over a dozen dog deaths have been attributed to cyanotoxin poisonings since 2000. During the summers of 2013-2015, we documented spatial and temporal patterns of cyanotoxin concentrations in the watershed, showing widespread distribution of anatoxin-a in benthic cyanobacterial mats. Solid phase adsorption toxin tracking (SPATT) samplers were deployed weekly to record dissolved microcystin and anatoxin-a levels at 10 sites throughout the watershed, and 187 Anabaena-dominated or Phormidium-dominated cyanobacterial mat samples were collected from 27 locations to measure intracellular anatoxin-a (ATX) and microcystins (MCY). Anatoxin-a levels were higher than microcystin for both SPATT (mean MCY = 0.8 and ATX = 4.8 ng g resin-1 day-1) and cyanobacterial mat samples (mean MCY = 0.074 and ATX = 1.89 μg g-1 DW). Of the benthic mats sampled, 58.9% had detectable anatoxin-a (max = 70.93 μg g-1 DW), while 37.6% had detectable microcystins (max = 2.29 μg g-1 DW). SPATT cyanotoxin levels peaked in mid-summer in warm mainstem reaches of the watershed. This is one of the first documentations of widespread anatoxin-a occurrence in benthic cyanobacterial mats in a North American watershed.

Topics: Anabaena; Animals; Bacterial Toxins; California; Cyanobacteria; Cyanobacteria Toxins; Dogs; Environmental Monitoring; Humans; Microcystins; Oscillatoria; Public Health; Rivers; Tropanes; Water Microbiology; Water Pollutants, Chemical; Water Quality

Benthic cyanobacteria in rivers produce cyanotoxins and affect aquatic food webs, but knowledge of their ecology lags behind planktonic cyanobacteria. The buoyancy of benthic Anabaena spp. mats was studied to understand implications for Anabaena dispersal in the Eel River, California. Field experiments were used to investigate the effects of oxygen bubble production and dissolution on the buoyancy of Anabaena dominated benthic mats in response to light exposure. Samples of Anabaena dominated mats were harvested from the South Fork Eel River and placed in settling columns to measure floating and sinking velocities, or deployed into in situ ambient and low light treatments to measure the effect of light on flotation. Floating and sinking occurred within minutes and were driven by oxygen bubbles produced during photosynthesis, rather than intracellular changes in carbohydrates or gas vesicles. Light experiment results showed that in a natural ambient light regime, mats remained floating for at least 4days, while in low light mats begin to sink in <24h. Floating Anabaena samples were collected from five sites in the watershed and found to contain the cyanotoxins anatoxin-a and microcystin, with higher concentrations of anatoxin-a (median 560, max 30,693ng/gDW) than microcystin (median 30, max 37ng/gDW). The ability of Anabaena mats to maintain their buoyancy will markedly increase their downstream dispersal distances. Increased buoyancy also allows toxin-containing mats to collect along shorelines, increasing threats to human and animal public health.

Topics: Anabaena; Bacterial Toxins; California; Cyanobacteria Toxins; Harmful Algal Bloom; Light; Microcystins; Population Dynamics; Rivers; Tropanes

Akinetes are resistant cells which have the ability to persist in sediment for several decades. We have investigated the temporal distribution of akinetes of two species,

Topics: Bacterial Toxins; Cyanobacteria; Cyanobacteria Toxins; Environmental Monitoring; Eutrophication; Genes, Bacterial; Geologic Sediments; History, 18th Century; History, 19th Century; History, 20th Century; History, 21st Century; Lakes; Microcystins; Tropanes; Water Pollution

Freshwater blooms of cyanobacteria (blue-green algae) in source waters are generally composed of several different strains with the capability to produce a variety of toxins. The major exposure routes for humans are direct contact with recreational waters and ingestion of drinking water not efficiently treated. The ultra high pressure liquid chromatography tandem mass spectrometry based analytical method presented here allows simultaneous analysis of 22 cyanotoxins from different toxin groups, including anatoxins, cylindrospermopsins, nodularin and microcystins in raw water and drinking water. The use of reference standards enables correct identification of toxins as well as precision of the quantification and due to matrix effects, recovery correction is required. The multi-toxin group method presented here, does not compromise sensitivity, despite the large number of analytes. The limit of quantification was set to 0.1 μg/L for 75% of the cyanotoxins in drinking water and 0.5 μg/L for all cyanotoxins in raw water, which is compliant with the WHO guidance value for microcystin-LR. The matrix effects experienced during analysis were reasonable for most analytes, considering the large volume injected into the mass spectrometer. The time of analysis, including lysing of cell bound toxins, is less than three hours. Furthermore, the method was tested in Swedish source waters and infiltration ponds resulting in evidence of presence of anatoxin, homo-anatoxin, cylindrospermopsin and several variants of microcystins for the first time in Sweden, proving its usefulness.

Topics: Alkaloids; Bacterial Toxins; Chemistry Techniques, Analytical; Chromatography, High Pressure Liquid; Cyanobacteria; Cyanobacteria Toxins; Drinking Water; Fresh Water; Humans; Marine Toxins; Microcystins; Sweden; Tandem Mass Spectrometry; Tropanes; Uracil

This work allowed assessing a widespread occurrence of Tychonema bourrellyi in the largest lakes south of the Alps (Garda, Iseo, Como and Maggiore). The taxonomy of the species was confirmed adopting a polyphasic approach, which included microscopic examinations, molecular (16S rRNA and rbcLX sequences) and (Lake Garda) ecological characterisations. Over 70% of the 36 isolates of Tychonema sampled from the four lakes tested positive for the presence of genes implicated in the biosynthesis of anatoxins (anaF and/or anaC) and for the production of anatoxin-a (ATX) and homoanatoxin-a (HTX). A detailed analysis carried out in Lake Garda showed strong ongoing changes in the cyanobacterial community, with populations of Tychonema developing with higher biovolumes compared to the microcystins (MCs) producer Planktothrix rubescens Moreover, the time × depth distribution of Tychonema was paralleled by a comparable distribution of ATX and HTX. The increasing importance of Tychonema in Lake Garda was also suggested by the opposite trends of ATX and MCs observed since 2009. These results suggest that radical changes are occurring in the largest lakes south of the Alps. Their verification and implications will require to be assessed by extending a complete experimental work to the other large perialpine lakes.

Topics: Bacterial Toxins; Bridged Bicyclo Compounds, Heterocyclic; Cyanobacteria; Cyanobacteria Toxins; Lakes; Microcystins; Phylogeny; RNA, Ribosomal, 16S; Switzerland; Tropanes

Due to changing global climatic conditions, a lot of attention has been given to cyanobacteria and their bioactive secondary metabolites. These conditions are expected to increase the frequency of cyanobacterial blooms, and consequently, the concentrations of cyanotoxins in aquatic ecosystems. Unfortunately, there are very few studies that address the effects of cyanotoxins on the physiology of phytoplankton species under different environmental conditions. In the present study, we investigated the effect of the cyanotoxin anatoxin-a (ATX-A) on Microcystis aeruginosa (cyanobacteria) and Acutodesmus acuminatus (chlorophyta) under varying light and nitrogen conditions. Low light (LL) and nitrogen limitation (LN) resulted in significant cell density reduction of the two species, while the effect of ATX-A on M. aeruginosa was not significant. However, under normal (NN) and high nitrogen (HN) concentrations, exposure to ATX-A resulted in significantly (p < 0.05) lower cell density of A. acuminatus. Pigment content of M. aeruginosa significantly (p < 0.05) declined in the presence of ATX-A, regardless of the light condition. Under each light condition, exposure to ATX-A caused a reduction in total microcystin (MC) content of M. aeruginosa. The detected MC levels varied as a function of nitrogen and ATX-A concentrations. The production of reactive oxygen species (H

Topics: Antioxidants; Cell Count; Cyanobacteria; Cyanobacteria Toxins; Hydrogen Peroxide; Light; Microcystins; Microcystis; Nitrogen; Phytoplankton; Scenedesmus; Tropanes

In the polar regions cyanobacteria are an important element of plant communities and represent the dominant group of primary producers. They commonly form thick highly diverse biological soil crusts that provide microhabitats for other organisms. Cyanobacteria are also producers of toxic secondary metabolites. In the present study we demonstrated that biocrust-forming cyanobacteria inhabiting the Kaffiøyra Plain, the north-west coast of Spitsbergen, are able to synthesize toxins, especially microcystins (MCs, from 0.123 to 11.058 μg MC-LR per g dry weight, DW) and anatoxin-a (ANTX-a, from 0.322 to 0.633 μg ANTX-a per g DW). To the best of our knowledge, this is the first report on the presence of ANTX-a in the entire polar region. The occurrence of cyanotoxins can exert a long-term impact on organisms co-existing in biocrust communities and can have far-reaching consequences for the entire polar ecosystem.

Topics: Arctic Regions; Biomass; Cyanobacteria; Cyanobacteria Toxins; Ecosystem; Fresh Water; Microcystins; Tropanes

Until recently, exposure pathways of concern for cyanotoxins have focused on recreational exposure, drinking water, and dermal contact. Exposure to cyanotoxins through fish consumption is a relatively new area of investigation. To address this concern, microcystins and other cyanotoxins were analyzed in fish collected from nine Washington lakes with recurrent toxic blooms using two types of enzyme-linked immunosorbent assays (ELISAs) and liquid chromatography/mass spectrometry/mass spectrometry (LC-MS/MS). Microcystins or microcystin-like compounds were elevated in fish liver relative to muscle and other tissues (liver>gut>muscle). Microcystin concentrations in fish fillet samples using anti-Adda ELISA (range 6.3-11 μg/kg wet weight) were consistently higher in all fish species than concentrations using anti-microcystin (MC)-leucine-arginine (LR) ELISA (range 0.25-2.4 μg/kg wet weight). MC-leucine-alanine (LA) was the only variant detected in fish (2.5-12 μg/kg in four livers) among the nine variants analyzed by LC-MS/MS. Fish fillets showed no accumulation of the MCs targeted by LC-MS/MS. Other cyanotoxins analyzed (anatoxin-a, saxitoxin, domoic acid, and okadaic acid) were not detected in fish. Based on this and evidence from other studies, we believe that people can safely consume two 8-oz fish fillet meals per week from lakes with blooms producing MCs (clean the fish and discard viscera).

Topics: Animals; Chromatography, Liquid; Cyanobacteria Toxins; Environmental Monitoring; Enzyme-Linked Immunosorbent Assay; Fishes; Lakes; Microcystins; Tandem Mass Spectrometry; Tropanes; Washington; Water Pollutants, Chemical

An analytical method based on on-line SPE-LC-HESI-MS/MS has been developed for the detection and quantification of eight selected cyanotoxins in algal bloom waters that include mycrocystins, anatoxin-a and cylindrospermopsin. The injection volume was 2 mL according to the expected concentration of cyanotoxins in matrix. The method provides an analysis time of 7 min per sample, acceptable recovery values (91-101%), good precision (RSD < 13%) and method limits of detection at the sub-microgram per liter levels (0.01-0.02 μg L(-1)). A detailed discussion on optimization parameters that have an impact on the overall performance of the method are presented. In particular, method optimization permitted the chromatographic separation of anatoxin-a and phenylalanine, an isobaric interference with a similar chromatographic characteristics. All optimization and validation experiments for the on-line SPE method and chromatographic separation were performed in environmentally relevant algal bloom water matrices. The applicability of the method was tested on several algal bloom water samples from monitored lakes across the province of Québec (Québec, Canada) known to produce cyanotoxins. All of the targeted cyanotoxins were detected with the exception of cylindrospermopsin. In addition, it was found that total microcystin concentrations in several surface water samples exceeded the proposed guidelines established by the province of Québec in Canada of 1.5 μg L(-1) as well as the World Health Organization of 1 μg L(-1) for both free and cell-bound microcystin-LR equivalent.

Topics: Alkaloids; Bacterial Toxins; Chromatography, Liquid; Cyanobacteria; Cyanobacteria Toxins; Environmental Monitoring; Harmful Algal Bloom; Marine Toxins; Microcystins; Solid Phase Extraction; Tandem Mass Spectrometry; Tropanes; Uracil

Several freshwater cyanobacteria species have the capability to produce toxic compounds, frequently referred to as cyanotoxins. The most prevalent of these cyanotoxins is microcystin LR. Recognizing the potential health risk, France, Italy, Poland, Australia, Canada, and Brazil have set either standards or guidelines for the amount of microcystin LR permissible in drinking water based on the World Health Organization guideline of one microg/L of microcystin LR. Recently, the United States Environmental Protection Agency has begun to evaluate the occurrence and health effects of cyanotoxins and their susceptibility to water treatment under the Safe Drinking Water Act through the Contaminant Candidate List (CCL). A recent update of the Contaminant Candidate List focuses research and data collection on the cyanotoxins microcystin LR, anatoxin-a, and cylindrospermopsin. Liquid Chromatography/Tandem-Mass Spectrometry (LC/MS/MS) is a powerful tool for the analysis of various analytes in a wide variety of matrices because of its sensitivity and selectivity. The use of smaller column media (sub 2 microm particles) was investigated to both improve the speed, sensitivity and resolution, and to quantify the CCL cyanotoxins, in a single analysis, using Ultra-Performance Liquid Chromatography (UPLC) combined with tandem mass spectrometry. Natural waters and spiked samples were analyzed to show proof-of-performance. The presented method was able to clearly resolve each of the cyanotoxins in less than eight minutes with specificity and high spike recoveries.

Topics: Alkaloids; Bacterial Toxins; Chromatography, High Pressure Liquid; Cyanobacteria; Cyanobacteria Toxins; Fresh Water; Marine Toxins; Microcystins; Saxitoxin; Spectrometry, Mass, Electrospray Ionization; Tandem Mass Spectrometry; Tropanes; Uracil; Water Pollutants, Chemical

Cyanobacterial toxins have adverse effects on both terrestrial and aquatic plants. Microcystins are cyclic heptapeptides and an important group of cyanotoxins. When lake water contaminated with cyanobacterial blooms is used for spray irrigation, these toxins can come in contact with agricultural plants. During the exposure to these toxins, reactive oxygen species can form. These reactive oxygen species have a strong reactivity and are able to interact with other cellular compounds (lipids, protein, and DNA). Plants have antioxidative systems that will limit the negative effects caused by reactive oxygen species. These systems consist of enzymes, such as superoxide dismutase, catalase, and ascorbate peroxidase, and nonenzymatic substances, such as reduced glutathione or vitamins. The aim of the present study was to investigate the effects of cyanobacterial toxins (microcystins and anatoxin-a) and cyanobacterial cell-free crude extract on alfalfa (Medicago sativa) seedlings. Inhibition of germination and root growth was observed with toxin concentrations of 5.0 microg/L. Also, oxidative damage, such as lipid peroxidation, was detected after the exposure of alfalfa seedlings to the toxin. Reactive oxygen detoxifying enzymes were elevated, showing a marked response in alfalfa to oxidative stress caused by the exposure to cyanobacterial metabolites that might influence the growth and development of these plants negatively.

Topics: Bacterial Toxins; Cyanobacteria; Cyanobacteria Toxins; Germination; Herbicides; Marine Toxins; Medicago sativa; Microcystins; Oxidative Stress; Paraquat; Peptides, Cyclic; Seeds; Tropanes

The first identification of anatoxin-a in a French lotic system is reported. Rapid deaths of dogs occurred in 2003 after the animals drank water from the shoreline of the La Loue River in eastern France. Sediments, stones and macrophytes surfaces at the margin of the river were covered by a thick biofilm containing large quantities of several benthic species of filamentous, non-heterocystous cyanobacteria. Known cyanotoxins, such as microcystins, saxitoxins and anatoxins were screened from biofilm samples by biochemical and analytical assays. A compound with similar UV spectra to the anatoxin-a standard was detected by high-performance liquid chromatography (HPLC) coupled with photo-diode array detector. This toxin was further identified by HPLC coupled with a UV detector and by electrospray ionisation-Quadrupole-Time-Of-Flight mass spectrometer, and confirmed by tandem mass spectrometry. These two techniques were necessary to discriminate anatoxin-a in phenylalanine-containing matrices such as liver samples of poisoned dogs. The toxin and the aromatic amino acid, phenylalanine, present the same pseudomolecular ion at m/z 166, but have differing fragmentation patterns, retention times and UV spectra. Finally, several cyanobacterial strains were isolated from the green biofilm and tested for anatoxin-a production. Phormidium favosum was identified as a new anatoxin-a producing species.

Topics: Animals; Bacterial Toxins; Cell Line, Tumor; Cyanobacteria; Cyanobacteria Toxins; Dogs; Environmental Monitoring; France; Gastrointestinal Contents; Intestines; Liver; Marine Toxins; Mice; Microcystins; Neurotoxins; Peptides, Cyclic; Poisoning; Rivers; Saxitoxin; Tropanes

Toxic and non-toxic cyanobacterial strains from Anabaena, Aphanizomenon, Calothrix, Cylindrospermum, Nostoc, Microcystis, Planktothrix (Oscillatoria agardhii), Oscillatoria and Synechococcus genera were examined by RFLP of PCR-amplified 16S rRNA genes and 16S rRNA gene sequencing. With both methods, high 16S rRNA gene similarity was found among planktic, anatoxin-a-producing Anabaena and non-toxic Aphanizomenon, microcystin-producing and non-toxic Microcystis, and microcystin-producing and non-toxic Planktothrix strains of different geographical origins. The respective sequence similarities were 99.9-100%, 94.2-99.9% and 99.3-100%. Thus the morphological characteristics (e.g. Anabaena and Aphanizomenon), the physiological (toxicity) characteristics or the geographical origins did not reflect the level of 16S rRNA gene relatedness of the closely related strains studied. In addition, cyanobacterial strains were fingerprinted with repetitive extragenic palindromic (REP)- and enterobacterial repetitive intergenic consensus (ERIC)-PCR. All the strains except two identical pairs of Microcystis strains had different band profiles. The overall grouping of the trees from the 16S rRNA gene and the REP- and ERIC-PCR analyses was similar. Based on the 16S rRNA gene sequence analysis, four major clades were formed. (i) The clade containing filamentous heterocystous cyanobacteria was divided into three discrete groups of Anabaena/Aphanizomenon, Anabaena/Cylindrospermum/ Nodularia/Nostoc and Calothrix strains. The three other clades contained (ii) filamentous non-heterocystous Planktothrix, (iii) unicellular non-heterocystous Microcystis and (iv) Synechococcus strains.

Topics: Anabaena; Animals; Bacterial Toxins; Cyanobacteria; Cyanobacteria Toxins; Marine Toxins; Microcystins; Molecular Sequence Data; Peptides, Cyclic; Phylogeny; Plankton; Polymorphism, Restriction Fragment Length; RNA, Ribosomal, 16S; Tropanes

Simple and easy-to-use bioassays with Artemia salina (brine shrimp) larvae, luminescent bacteria and Pseudomonas putida were evaluated for the detection of toxicity due to cyanobacterial hepato- and neurotoxins. The hepatotoxins and a neurotoxin, anatoxin-a, were extracted from laboratory-grown cultures and natural bloom samples by the solid phase fractionation method and dissolved in diluent for different bioassays. The toxin concentration of cyanobacterial extracts was determined with HPLC. The Artemia biotest appeared to be quite sensitive to cyanobacterial hepatotoxins, with LC 50 values of 3-17 mg l-1. The Artemia test was also shown to be of value for the detection of toxicity caused by anatoxin-a. The fractionated extract of anatoxin-a was not lethal to Artemia but it disturbed the ability of the larvae to move forwards. Filtered cyanobacterial cultures with anatoxin-a, on the other hand, caused mortality of Artemia larvae at concentrations of 2-14 mg l-1. With the solid phase fractionation of cyanobacterial samples, no non-specific toxicity due to compounds other than hepato- and neurotoxins was observed. In the luminescent bacteria test, the inhibition of luminescence did not correlate with the abundance of hepatotoxins or anatoxin-a. The growth of Ps. putida was enhanced, rather than inhibited by cyanobacterial toxin fractions.

Topics: Animals; Artemia; Bacterial Toxins; Cyanobacteria; Cyanobacteria Toxins; Marine Toxins; Microcystins; Peptides, Cyclic; Photobacterium; Pseudomonas putida; Tropanes; Vibrio