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The aim of the study was to determine the toxicity of the extract obtained from the cyanobacterial cells derived from the waters of Zemborzycki dam reservoir with use of a battery of biotests. The taxonomic identification of the bloom-forming cyanobacteria revealed high abundance of Aphanizomenon flos-aquae and Dolichospermum spp. (Anabaena spp.) and in a lower degree of Microcystis aeruginosa and Planktothrix agardhii. In the extract obtained from concentrated cyanobacterial cells, hepatotoxin microcystin-LR at a concentration of 22.89 ± 3.74 μg/L and neurotoxin Antx-a at 13.02 ± 0.01 μg/L have been detected. Toxicity of the extract was evaluated with the following assays: Daphtoxkit F magna with the crustacean Daphnia magna, Thamnotoxkit F with the crustacean Thamnocephalus platyurus, Rotoxkit F with the rotifer Brachionus calyciflorus and Protoxkit F with ciliate Tetrahymena thermophila. The most sensitive organism among all studied was T. platyurus for which EC50 was estimated to be 1.2% of the initial extract concentration. On the basis of the highest obtained value of the toxicity unit (TU = 83) the studied sample was classified to the IV class, which is of high acute toxicity. Additionally, it was found that reactivity on cyanobacterial products differs greatly among organisms used in bioassays, which indicate the need for using a set of biotests.

Microcystin-LR (MC-LR) is a growing issue as it is toxic and difficult to remove in drinking water treatment plants (DWTPs). Mesoporous carbon (MC) is evaluated as an alternative adsorbent for MC-LR removal and compared with three widely-used powdered activated carbons (PACs). MC was more favorable for MC-LR removal than PACs. MC-LR adsorption on MC was a rapid process ( k 2 = 1.02 × 10 −4 g/μg/min) that completed within 15 min, while adsorption on PACs took 60 min. The maximum adsorption capacity of MC-LR was 18,008 μg/g (MC), which was higher than that of the PACs. Two mechanisms were associated with adsorption: the small hydro-dynamic diameter of MC in an aqueous solution increased the instantaneous attraction of MC-LR to its surface, and the numerous mesopores enhanced pore diffusion. The MC could remove MC-LR to meet the drinking water guidance level (1 μg/L) from an the MC-LR concentration range of 5–20 μg/L in drinking water sources, and 10 min of treatment was sufficient to meet this level (MC dose = 20 mg/L). The field-scale DWTP was operated by adding 1 or 5 mg/L MC to the mixing basin, and 49.49% and 74.50% of MC-LR was removed, respectively. Geosmin and 2-methylisoborneol were slightly reduced when 5 mg/L of MC was applied. [ABSTRACT FROM AUTHOR]

Despite the importance of understanding the bloom mechanisms that influence cyanobacterial toxin production, the dynamics of toxic Microcystis subpopulations are largely unknown. Here, we quantified both toxic and entire (i.e., toxic and non-toxic) Microcystis populations based on the microcystin synthetase E ( mcyE) and 16S ribosomal RNA genes. Samples were collected from pelagic water and sediments twice per week from October to December 2011, and we investigated the effects of physicochemical factors (pH, water temperature, dissolved oxygen, nutrients, etc.) and biological factors (ciliates and zooplankton) on the abundance of toxic and non-toxic Microcystis. During the study period, Microcystis blooms were composed of toxic and non-toxic subpopulations. Resting stage Microcystis in sediment may be closely linked to Microcystis populations in pelagic water and may contribute to the toxic subpopulation composition in surface Microcystis blooms. In pelagic water, the toxic and entire Microcystis population had a significant positive correlation with the pH and water temperature ( p < 0.05). However, their responses to changes in environmental factors were thought to be distinct. The ratio of the toxic to non-toxic Microcystis subpopulations was significantly ( p < 0.05) enhanced by a lower pH and water temperature and an increase in protozoan grazers, reflecting environmental stresses. These results suggest that the toxic and non-toxic subpopulations of Microcystis have distinct tolerance levels against these stressors. The intracellular microcystin (MC) concentration was positively associated with the abundance of the mcyE-positive Microcystis. By comparison, the MC concentration in pelagic water body (extracellular) increased when Microcystis was lysed due to environmental stresses. [ABSTRACT FROM AUTHOR]

Based on field measurements of water surface reflectance spectra in Lake Taihu, we construct a model for distinguishing between cyanobacterial bloom and floating leaf vegetation by combining a chlorophyll spectral index with a baseline of phycocyanin. In situ $R_{{\rm{rs}}}$ measurements validation results show that this model performs well in distinguishing cyanobacterial bloom from floating leaf vegetation in Lake Taihu. We apply this model to 52 remote sensing images from the Medium-Resolution Imaging Spectrometer (MERIS) from 2003 to 2011. Using two different accuracy evaluation methods, we find an average recognition accuracy of more than 80% for cyanobacterial bloom and floating leaf vegetation when using optimal index thresholds. Using an average index threshold to extract cyanobacterial bloom and floating leaf vegetation from the images, the relative accuracies are 78.8% and 74.6%, respectively. If more efficiency is desired, these average thresholds can be used, which is convenient for batch processing and automated extraction of cyanobacterial bloom and floating leaf vegetation from remote sensing data. The overall distribution of cyanobacterial bloom and floating leaf vegetation in Lake Taihu from 2003 to 2011 is determined by overlapping the distribution maps from individual images, and the results of our analysis are consistent with previously published results. In addition, our analysis shows that this model is immune to perturbations from thin clouds and aerosols. [ABSTRACT FROM PUBLISHER]

Monitoring of water and surface sediment in a French eutrophic lake (Lake Aydat) was carried out over a 2-year period in order to determine whether akinetes in sediment could be representative of the most recent bloom and to estimate their germination potential. Sediment analysis revealed two akinete species, Dolichospermum macrosporum and Dolichospermum flos-aquae, present in the same proportions as observed for the pelagic populations. Moreover, similar spatial patterns observed for vegetative cells in the water column and akinete distributions in the sediment suggest that akinetes in the sediment may be representative of the previous bloom. However, the relationship between akinetes in the sediment and vegetative cells in the water column was not linear, and other factors may interfere. For example, our results highlighted horizontal transport of akinetes during the winter. The benthic overwinter phase did not seem to influence the percentages of intact akinetes, which remained stable at approximately 7% and 60% for D. macrosporum and D. flos-aquae, respectively. These percentages may thus be the result of processes that occurred in the water column. The intact overwintering akinetes showed germination rates of up to 90% after 72 h for D. flos-aquae or 144 h for D. macrosporum. The difference in akinete germination rates between these two species demonstrates different ecological strategies, which serve to expand the window for germination in time and space and thus optimize colonization of the water column by nostocalean cyanobacteria. [ABSTRACT FROM AUTHOR]

Cyanobacterial blooms often occur in aquaculture wastewater in China. A floating plant, water hyacinth has been widely used to treat this wastewater. Little is known, however, about bacterial community characteristics and the risk of potential pathogens in cyanobacteria-blooming aquaculture wastewater remediated by water hyacinth. In wastewater treated with water hyacinth, we used culture enumeration and high-throughput sequencing to explore the characteristics of bacterial communities, the status of coliform bacteria, and pathogenic bacteria potentially conducive to human disease. Our results indicated that the relative abundance of Acidobacteria, Planctomycetes, Actinobacteria, Chlorobi, Cyanobacteria, Proteobacteria, and phylum OD1 in cyanobacteria-blooming aquaculture wastewater were significantly influenced by water hyacinth. After 30 days, the relative abundance of Proteobacteria and phylum OD1 in the water hyacinth treatments increased remarkably, while the relative abundance of the other 5 phyla in treatment was significantly reduced compared with the controls. In 21 major families, the relative abundance of Comamonadaceae, Oxalobacteraceae, Rhodocylclaceae, and an unnamed group from phylum OD1 increased significantly in the water hyacinth treatments compared with the controls. The number of total coliforms in wastewater treated by water hyacinth was significantly elevated and higher than controls during the first 6-18 days, with the maximum reaching 23,800 MPN/L. The level of potential pathogenic bacteria in wastewater treated by water hyacinth significantly reduced compared with the controls after 18 days, but it significantly increased from the initial level. It appears that water hyacinth by itself is not an effective treatment for reducing potential pathogens in aquaculture water. [ABSTRACT FROM AUTHOR]

The degradation of freshwater quality induced by cyanobacterial blooms is a major global environmental concern. Microbially driven nitrogen removal could alleviate eutrophication to some degree in freshwater ecosystems. However, the response of anaerobic ammonium oxidizing (anammox) bacterial communities to cyanobacterial blooms remains poorly understood, especially in reservoir ecosystems. Here we compared the dynamics of anammox bacterial communities during and after a cyanobacterial bloom in a subtropical reservoir. Our data showed that a cyanobacterial bloom triggered a significant increase in bottom anammox bacterial abundance. During the bloom period, anammox bacterial abundance in bottom waters was 9-fold and 52-fold higher compared with non-bloom stratification and mixing periods, respectively. The community composition of anammox bacteria in surface waters changed substantially accompanied by the disappearance of the cyanobacterial bloom, and a shift of dominance from unidentified anammox genera to Ca. Brocadia was observed. Although Ca. Brocadia was always predominant in both middle and bottom waters, the non-bloom period had more unique taxa than the bloom period. Cyanobacterial bloom-related changes in environmental conditions (e.g. NH4-N and total organic carbon) and water stratification together influenced the distribution and dynamics of anammox bacteria. Altogether, our study lays the basis for a better understanding of the breakdown of cyanobacterial blooms in a stratified reservoir. [ABSTRACT FROM AUTHOR]

Many countries in the world still suffer from high toxic cyanobacterial blooms in inland waters used for human consumption. Regional climate change and human activities within watersheds exert a complex and diverse influence on aquatic ecosystem structure and function across space and time. However, the degree to which these factors may contribute to the long-term dynamics of plankton communities is still not well understood. Here, we explore the impacts of multiple disturbance events (e.g. human-resettlement, temperature change, rainfall, water level fluctuations), including six combined disturbances, on phytoplankton and cyanobacteria in two subtropical reservoirs over six years. Our data showed that combined environmental disturbances triggered two apparent and abrupt switches between cyanobacteria-dominated state and non-cyanobacterial taxa-dominated state. In late 2010, the combined effect of human-resettlement (emigration) and natural disturbances (e.g. cooling, rainfall, water level fluctuations) lead to a 60–90% decrease in cyanobacteria biomass accompanied by the disappearance of cyanobacterial blooms, in tandem with an abrupt and persistent shift in phytoplankton community. After summer 2014, however, combined weather and hydrological disturbances (e.g. warming, rainfall, water level fluctuations) occurred leading to an abrupt and marked increase of cyanobacteria biomass, associated with a return to cyanobacteria dominance. These changes in phytoplankton community were strongly related to the nutrient concentrations and water level fluctuations, as well as water temperature and rainfall. As both extreme weather events and human disturbances are predicted to become more frequent and severe during the twenty-first century, prudent sustainable management will require consideration of the background limnologic conditions and the frequency of disturbance events when assessing the potential impacts on reservoir biodiversity and ecosystem functioning and services. [ABSTRACT FROM AUTHOR]

In this study, the distribution, transfer and fate of both polychlorinated biphenyls (PCBs) and cyanotoxins via phytoplankton routes were systematically investigated in two Chinese lakes. Results indicated that PCB adsorption/bioaccumulation dynamics has significantly positive correlations with the biomass of green alga and diatoms. Total lipid content of phytoplankton is the major factor that influences PCB adsorption/bioaccumulation. Cyanobacterial blooms with relatively lower lipid content could also absorb high amount of PCBs due to their high cell density in the water columns, and this process was proposed as major route for the transfer of PCBs in Chinese eutrophic freshwater. According to these findings, a novel route on fates of PCBs via phytoplankton and a green bioadsorption concept were proposed and confirmed. In the practice of mechanical collections of bloom biomass from Lake Taihu, cyanotoxin/cyanobacteria and PCBs were found to be removed simultaneously very efficiently followed this theory. [ABSTRACT FROM AUTHOR]

An enclosure experiment was conducted between July and September 2009 to compare the effectiveness of a phosphate fixative, the lanthanum-modified bentonite clay Phoslock® (LMB), dredging, and their combination in controlling eutrophication in a hypertrophic urban pond in Heesch, The Netherlands. In total, 25 water quality variables were monitored. Multivariate analysis revealed that the combination LMB-treated and dredged enclosures deviated most from the pond (reference) and the controls, and showed the strongest eutrophication reduction. Overall, dredging significantly increased transparency, lowered turbidity, and improved the oxygen conditions in the enclosures compared to non-dredged ones. Nonetheless, one dredged enclosure deviated dramatically from the others, which might reflect methodological issues with dredging. The LMB treatment appeared to be less effective at mitigating eutrophication than dredging, and phosphate concentrations even increased during the experiment in the LMB-treated enclosures. Chemical equilibrium modeling suggested that humic substances could have formed complexes with lanthanum (La) from the LMB, rendering it unavailable for intercepting P over the course of the enclosure experiment. Residual lanthanum concentrations in combination dredging and LMB treatments exceeded the Dutch standard 10-fold. Total zooplankton abundance, and particularly Cladocera, increased in all enclosures over the course of the experiment. The limited effect of LMB in the enclosure experiment and the violation of the Dutch La standard when combined with dredging disqualify LMB as an intervention agent in the restoration of the pond. [ABSTRACT FROM AUTHOR]

Understanding of the bacterial community structure in drinking water resources helps to enhance the security of municipal water supplies. In this study, bacterial communities were surveyed in water and sediment during a heavy cyanobacterial bloom in a drinking water resource of Lake Taihu, China. A total of 325,317 high-quality sequences were obtained from different 16S ribosomal RNA (rRNA) regions (V3, V4, and V6) using the Miseq sequencing platform. A notable difference was shown between the water and sediment samples, as predominated by Cyanobacteria, Proteobacteria, and Actinobacteria in the water and Proteobacteria, Chloroflexi, and Verrucomicrobia in the sediment, respectively. The LD12 family dominated the water surface and was tightly associated with related indicators of cyanobacterial propagation, indicating involvement in the massive proliferation of cyanobacterial blooms. Alternatively, the genus Nitrospira dominated the sediment samples, which indicates that nitrite oxidation was very active in the sediment. Although pathogenic bacteria were not detected in a large amount, some genera such as Mycobacterium, Acinetobacter, and Legionella were still identified but in very low abundance. In addition, the effects of different V regions on bacterial diversity survey were evaluated. Overall, V4 and V3 were proven to be more promising V regions for bacterial diversity survey in water and sediment samples during heavy water blooms in Lake Taihu, respectively. As longer, cheaper, and faster DNA sequencing technologies become more accessible, we expect that bacterial community structures based on 16S rRNA amplicons as an indicator could be used alongside with physical and chemical indicators, to conduct comprehensive assessments for drinking water resource management. [ABSTRACT FROM AUTHOR]

The issues presented in this study concern the important problem of the occurrence of cyanobacterial blooms in surface water used for water supply purposes. The objective of this study was to analyze the occurrence of cyanotoxic risk in the catchment area of the Obrzyca River, which is a source of drinking water for the inhabitants of Zielona Góra. At the points of Uście and at its tributary at Wojnowo, the river was more exposed to the blooms of toxigenic algae, whose active microcystins synthesize more here than in other places (Sadowo, Chwalim, Ostrzyce, Lubiatów). Throughout the study, conducted in 2008-12, we observed several times that the amounts of cyanobacteria exceeded 500·103 individuals per litre (ind.·dm-3 ). The observed cyanobacterial blooms were dominated by the following species: Dolichospermum flos-aquae and Planktothrix agardhii in the Obrzyca River at Uście, and D. spiroides, D. affinis, and Aphanizomenon flos-aquae at its tributary in Wojnowo. The maximum value of intracellular MC-LR, totalling 21.4 μg·dm-3 , was found at Wojnowo in September 2011. The analysis of the physico-chemical water quality indicators showed that the places along the Obrzyca River where periodic cyanobacterial blooms took place were characterized by eutrophic conditions, and the total nitrogen to total phosphorus ratio (N/P) fell within the range of 10 to 16, and the water temperature exceeded 20ºC. In order to assess the toxic hazard caused by periodic cyanobacterial blooms in the catchment area of the Obrzyca River for drinking water for Zielona Góra residents, regular monitoring of the cyanobacteria and their toxins is required – especially at the points in Uście and Wojnowo as well as at the water intake at the water treatment plant (WTP) in Zawada (in central-western Poland). [ABSTRACT FROM AUTHOR]

Cyanobacterial blooms in eutrophic lakes are severe environmental problems worldwide. To characterize the spatiotemporal heterogeneity of cyanobacterial blooms, a high-throughput method is necessary for the specific detection of cyanobacteria. In this study, the cyanobacterial composition of three eutrophic waters in China (Taihu Lake, Dongqian Lake, and Dongzhen Reservoir) was determined by pyrosequencing the cpcBA intergenic spacer ( cpcBA -IGS) of cyanobacteria. A total of 2585 OTUs were obtained from the normalized cpcBA -IGS sequence dataset at a distance of 0.05. The 238 most abundant OTUs contained 92% of the total sequences and were classified into six cyanobacterial groups. The water samples of Taihu Lake were dominated by Microcystis , mixed Nostocales species, Synechococcus , and unclassified cyanobacteria. Besides, all the samples from Taihu Lake were clustered together in the dendrogram based on shared abundant OTUs. The cyanobacterial diversity in Dongqian Lake was dramatically decreased after sediment dredging and Synechococcus became exclusively dominant in this lake. The genus Synechococcus was also dominant in the surface water of Dongzhen Reservoir, while phylogenetically diverse cyanobacteria coexisted at a depth of 10 m in this reservoir. In summary, targeted deep sequencing based on cpcBA -IGS revealed a large diversity of bloom-forming cyanobacteria in eutrophic lakes and spatiotemporal changes in the composition of cyanobacterial communities. The genus Microcystis was the most abundant bloom-forming cyanobacteria in eutrophic lakes, while Synechococcus could be exclusively dominant under appropriate environmental conditions. [ABSTRACT FROM AUTHOR]

Research on the combined effects of antibiotic contaminants and environmental factors in cyanobacteria is still limited. This study focused on the action and its mechanism of spiramycin combined with changes in nitrogen and phosphorus level in Microcystis aeruginosa at environmentally relevant concentrations. Though photosynthetic activity was stimulated by spiramycin at a high nutrient level, no significant correlation ( p > 0.05) was found between photosynthesis-related proteins and growth-related proteins, and the growth rate was inhibited by 200 ng L −1 of spiramycin. At low nitrogen and low phosphorus levels, up-regulated photosynthesis-related proteins were closely correlated with ( p < 0.05) stress response-related, transcription-related and cell division-related proteins, which consequently led to stimulated growth of M. aeruginosa under spiramycin exposure. Spiramycin exposure also regulated the production of microcystins (MCs) and the expression of two microcystin synthetases (mcyB and mcyC). The spiramycin-induced protein secretion process and the up-regulation of ATP binding cassette transporters might contribute to the increased MC release. Enolase, superoxide dismutase, protein GrpE, DNA-directed RNA polymerase subunit alpha and serine protease were candidate target proteins of spiramycin in M. aeruginosa under different nutrient conditions. Coexisting spiramycin mitigated the threat of cyanobacteria to aquatic environments at a high nutrient level but aggravated cyanobacterial bloom at a low nitrogen level. [ABSTRACT FROM AUTHOR]

Increased nutrient enrichment in Mediterranean standing waters has enhanced the risk of being affected by cyanobacterial blooms. Because phosphorus abatement is shaped as a crucial strategy for controlling eutrophication, this study introduces a structural thinking, experiential learning laboratory with animation dynamic model elaborated for Cazalegas Reservoir (Spain) to assess the feasibility of implementing a set of internal and external control measures and hydromorphological adjustments to meet the goal of oligotrophication. This shallow reservoir is another case where recurrent eutrophication has led to reach annual mean total phosphorus concentrations (0.16 ± 0.08 mg total phosphorus/L) over the threshold of current water policies, triggering cyanobacterial growth up to undesirable levels in summer time (approximately 50,000 cells/mL). Modeling results showed that (i) after upgrading water treatment in the main tributary, (ii) applying a lanthanum-modified bentonite into the water column and sediment, and (iii) increasing reservoir water level, in-lake P concentrations and cyanobacterial abundance decreased in an 88% (below 0.01 mg total phosphorus/L) and 84% (below 6000 cells/mL), respectively in the most critical periods. However, the constraints of the proposed management strategies are associated with their costs of implementation and the time span for a stable trophic recovery of the reservoir. In that end, integrated management approaches are aimed to be adopted by water managers to reach adequate ecological status of freshwater bodies. [ABSTRACT FROM AUTHOR]

The cyanobacterium Aphanizomenon favaloroi was identified for the first time in Europe in the Mediterranean brackish Lake Vistonis during July-August 2014. It formed a dense bloom (from 16×106 to 81×106 trichomes L-1), causing a brown colouration of the lake water. When A. favaloroi formed 100% of the total phytoplankton biomass (44mg L -1), saxitoxins (saxitoxin and neo-saxitoxin) were detected in the lake seston (42 and 17 μgg-1 phytoplankton dry weight respectively), which was screened for saxitoxins, cylindrospermopsin, anatoxin-a, microcystins and nodularin. A massive fish kill coincided with the A. favaloroi bloom. This new saxitoxin-producing species of cyanobacteria, with traits for successful dispersal, may pose a health risk to animals and humans and cause adverse effects on water quality and water services because of its expansion potential. [ABSTRACT FROM AUTHOR]

• High frequency monitoring of phytoplankton and bacterioplankton dynamics in an eutrophic shallow lake. • Phytoplankton and associated bacterioplankton communities can be seen as a couple in interaction but with flexible species variations. • Free-living bacterioplankton is under less direct influence of the phytoplankton communities and environmental variables dynamics. Bloom-forming phytoplankton dynamics are still unpredictable, even though it is known that several abiotic factors, such as nutrient availability and temperature, are key factors for bloom development. We investigated whether biotic factors, i.e. the bacterioplankton composition (via 16SrDNA metabarcoding), were correlated with phytoplankton dynamics, through a weekly monitoring of a shallow lake known to host recurrent cyanobacterial blooms. We detected concomitant changes in both bacterial and phytoplankton community biomass and diversity. During the bloom event, a significant decrease in phytoplankton diversity, was detected, with a first co-dominance of Ceratium, Microcystis and Aphanizomenon , followed by a co-dominance of the two cyanobacterial genera. In the same time, we observed a decrease of the particle-associated (PA) bacterial richness and the emergence of a specific bacterial consortium that was potentially better adapted to the new nutritional niche. Unexpectedly, changes in PA bacterial communities occurred just before the development the emergence of the phytoplanktonic bloom and the associated modification of the phytoplanktonic community composition, suggesting that changes in environmental conditions leading to the bloom, were first sensed by the bacterial PA community. This last was quite stable throughout the bloom event, even though there were changes in the blooming species, suggesting that the association between cyanobacterial species and bacterial communities may not be as tight as previously described for monospecific blooming communities. Finally, the dynamics of the free-living (FL) bacterial communities displayed a different trajectory from those of the PA and phytoplankton communities. This FL communities can be viewed as a reservoir for bacterial recruitment for the PA fraction. Altogether, these data also highlight s that the spatial organization within these different microenvironments in the water column is a relevant factor in the structuring of these communities. [ABSTRACT FROM AUTHOR]

The three Gram-negative, catalase- and oxidase-positive bacterial strains RS43 T , HBC28, and HBC61 T , were isolated from fresh water and subjected to a polyphasic study. Comparison of 16S rRNA gene sequence initially indicated that strains RS43 T , HBC28, and HBC61 T were closely related to species of genus Curvibacter and shared the highest sequence similarity of 98.14%, 98.21%, and 98.76%, respectively, with Curvibacter gracilis 7-1 T . Phylogenetic analysis based on genome sequences placed all strains within the genus Curvibacter . The average nucleotide identity (ANI) and digital DNA-DNA hybridization (dDDH) values between the three strains and related type strains supported their recognition as two novel genospecies in the genus Curvibacter . Comparative genomic analysis revealed that the genus possessed an open pangenome. Based on KEGG BlastKOALA analyses, Curvibacter species have the potential to metabolize benzoate, phenylacetate, catechol, and salicylate, indicating their potential use in the elimination of these compounds from the water systems. The results of polyphasic characterization indicated that strain RS43 T and HBC61 T represent two novel species, for which the name Curvibacter microcysteis sp. nov. (type strain RS43 T =KCTC 92793 T =LMG 32714 T ) and Curvibacter cyanobacteriorum sp. nov. (type strain HBC61 T =KCTC 92794 T =LMG 32713 T ) are proposed.

Microcystins (MCs) are the most frequently found cyanobacterial toxins in freshwater systems. Many MC variants have been identified and variants differ in their toxicity. Recent studies showed that the variants MC-LW and MC-LF might be more toxic than MC-LR, the variant that is most abundant and mostly used for risk assessments. As little is known about the presence of these two variants in The Netherlands, we determined their occurrence by analyzing 88 water samples and 10 scum samples for eight MC variants ((dm-7-)MC-RR, MC-YR, (dm-7-)MC-LR, MC-LY, MC-LW and MC-LF) by liquid chromatography with tandem mass spectrometry detection. All analyzed MC variants were detected, and MC-LW and/or MC-LF were present in 32% of the MC containing water samples. When MC-LW and MC-LF were present, they contributed to nearly 10% of the total MC concentrations, but due to their suspected high toxicity, their average contribution to the total MC toxicity was estimated to be at least 45%. Given the frequent occurrence and possible high toxicity of MC-LW and MC-LF, it seems better to base health risk assessments on the toxicity contributions of different MC variants than on MC-LR concentrations alone.