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The authors acknowledge COST Action ES 1105 "CYANOCOST Cyanobacterial blooms and toxins in water resources: Occurrence impacts and management" and COST Action Global Change Biology ES 1201 NETLAKE -Networking Lake Observatories in Europe" for contributing to this study through networking and knowledge sharing with European experts in the field. We acknowledge the members of the Global Lake Ecological Observatory Network (GLEON) for their collaborative spirit and enthusiasm that inspired the grassroots effort of the EMLS. E.M. was supported by a grant from the Swiss State Secretariat for Education, Research and Innovation to Bas Ibelings and by supplementary funding from University of Geneva. We thank Wendy Beekman for the nutrient analysis. We thank Pieter Slot for assisting with the pigment analysis. We thank Dr. Ian Jones for valuable feedback on an earlier version of the manuscript. We thank the Leibniz Institute of Freshwater Ecology and the Aquatic Microbial Ecology Group for logistic and technical support of J. Fonvielle and H.-P. Grossart, and the Leibniz Association for financial support. H.P. was supported by the US National Science Foundation (1840715, 1831096). A.C.'s work was funded by the Spanish Agencia Estatal de Investigacion and EU funds through the project CLIMAWET (CGL2015-69557-R). The collection of data for Lough Erne and Lough Neagh were funded by the Department of Agriculture, Environment and Rural Affairs, Northern Ireland. We are grateful to Kristiina Vuorio from the Freshwater Centre of the Finnish Environment institute for her help in organizing, collecting and analysing samples by the University of Jyvaskyla and to Gerald Dorflinger from the Water Development Department of Cyprus for his assistance with the sampling in Cyprus and for granting the CUT team permission to use WDD's equipment. Finally, we would like to thank the numerous other assistants that helped realizing each local survey. Open access funding provided by Universite de Geneve., To determine the drivers of phytoplankton biomass, we collected standardized morphometric, physical, and biological data in 230 lakes across the Mediterranean, Continental, and Boreal climatic zones of the European continent. Multilinear regression models tested on this snapshot of mostly eutrophic lakes (median total phosphorus [TP] = 0.06 and total nitrogen [TN] = 0.7 mg L-1), and its subsets (2 depth types and 3 climatic zones), show that light climate and stratification strength were the most significant explanatory variables for chlorophyll a (Chl a) variance. TN was a significant predictor for phytoplankton biomass for shallow and continental lakes, while TP never appeared as an explanatory variable, suggesting that under high TP, light, which partially controls stratification strength, becomes limiting for phytoplankton development. Mediterranean lakes were the warmest yet most weakly stratified and had significantly less Chl a than Boreal lakes, where the temperature anomaly from the long-term average, during a summer heatwave was the highest (+4 degrees C) and showed a significant, exponential relationship with stratification strength. This European survey represents a summer snapshot of phytoplankton biomass and its drivers, and lends support that light and stratification metrics, which are both affected by climate change, are better predictors for phytoplankton biomass in nutrient-rich lakes than nutrient concentrations and surface temperature., European Cooperation in Science and Technology (COST) ES 1105 ES 1201, Swiss State Secretariat for Education, Research and Innovation, University of Geneva, Leibniz Association, National Science Foundation (NSF) 1840715 1831096, Spanish Agencia Estatal de Investigacion, European Commission CGL2015-69557-R, Department of Agriculture, Environment and Rural Affairs, Northern Ireland

Insight into how environmental change determines the production and distribution of cyanobacterial toxins is necessary for risk assessment. Management guidelines currently focus on hepatotoxins (microcystins). Increasing attention is given to other classes, such as neurotoxins (e.g., anatoxin-a) and cytotoxins (e.g., cylindrospermopsin) due to their potency. Most studies examine the relationship between individual toxin variants and environmental factors, such as nutrients, temperature and light. In summer 2015, we collected samples across Europe to investigate the effect of nutrient and temperature gradients on the variability of toxin production at a continental scale. Direct and indirect effects of temperature were the main drivers of the spatial distribution in the toxins produced by the cyanobacterial community, the toxin concentrations and toxin quota. Generalized linear models showed that a Toxin Diversity Index (TDI) increased with latitude, while it decreased with water stability. Increases in TDI were explained through a significant increase in toxin variants such as MC-YR, anatoxin and cylindrospermopsin, accompanied by a decreasing presence of MC-LR. While global warming continues, the direct and indirect effects of increased lake temperatures will drive changes in the distribution of cyanobacterial toxins in Europe, potentially promoting selection of a few highly toxic species or strains. © 2018 by the authors. Licensee MDPI, Basel, Switzerland., The authors acknowledge COST Action ES 1105 “CYANOCOST—Cyanobacterial blooms and toxins in water resources: Occurrence impacts and management” and COST Action ES 1201 “NETLAKE—Networking Lake Observatories in Europe” for contributing to this study through networking and knowledge sharing with European experts in the field. Evanthia Mantzouki was supported by a grant from the Swiss State Secretariat for Education, Research and Innovation (SERI) to Bas Ibelings and by supplementary funding from the University of Geneva. We thank Clare Ahnlund, Ena Suarez and Irene Gallego for helping out with the Swiss survey. We thank Wendy Beekman and Els J. Faassen for the nutrient and toxin analysis.

Under ongoing climate change and increasing anthropogenic activity, which continuously challenge ecosystem resilience, an in-depth understanding of ecological processes is urgently needed. Lakes, as providers of numerous ecosystem services, face multiple stressors that threaten their functioning. Harmful cyanobacterial blooms are a persistent problem resulting from nutrient pollution and climate-change induced stressors, like poor transparency, increased water temperature and enhanced stratification. Consistency in data collection and analysis methods is necessary to achieve fully comparable datasets and for statistical validity, avoiding issues linked to disparate data sources. The European Multi Lake Survey (EMLS) in summer 2015 was an initiative among scientists from 27 countries to collect and analyse lake physical, chemical and biological variables in a fully standardized manner. This database includes in-situ lake variables along with nutrient, pigment and cyanotoxin data of 369 lakes in Europe, which were centrally analysed in dedicated laboratories. Publishing the EMLS methods and dataset might inspire similar initiatives to study across large geographic areas that will contribute to better understanding lake responses in a changing environment., The authors acknowledge COST Action ES 1105 “CYANOCOST – Cyanobacterial blooms and toxins in water resources: Occurrence impacts and management” and COST Action ES 1201 “NETLAKE – Networking Lake Observatories in Europe” for contributing to this study through networking and knowledge sharing with European experts in the field. Evanthia Mantzouki was supported by a grant from the Swiss State Secretariat for Education, Research and Innovation (SERI) to Bas Ibelings and by supplementary funding from University of Geneva. We thank Wendy Beekman for the nutrient analysis and the University of Wageningen for covering the costs of this analysis from the personal funding of dr. Miquel Lürling. We thank Pieter Slot for assisting with the pigment analysis and the University of Amsterdam for covering the costs of the analysis through funding from the group of Prof. Jef Huisman and dr. Petra Visser (IBED). We would like to thank the Environmental Data Initiative for covering the cost of archiving the EMLS dataset. We would like to thank the Leibniz Institute of Freshwater Ecology and Inland Fisheries, Dept. of Experimental Limnology and the Aquatic Microbial Ecology Group for logistic and technical support of J. Fonvielle and H.-P. Grossart; and the Leibniz Association for financial support. The collection of data for Lough Erne and Lough Neagh were funded by the Department of Agriculture, Environment and Rural Affairs, Northern Ireland.

Blooms of toxic cyanobacteria are becoming increasingly frequent, mainly due to water quality degradation. This work applied qPCR as a tool for early warning of microcystin(MC)-producer cyanobacteria and risk assessment of water supplies. Specific marker genes for cyanobacteria, Microcystis and MC-producing Microcystis, were quantified to determine the genotypic composition of the natural Microcystis population. Correlations between limnological parameters, pH, water temperature, dissolved oxygen and conductivity and MC concentrations as well as Microcystis abundance were assessed. A negative significant correlation was observed between toxic (with mcy genes) to non-toxic (without mcy genes) genotypes ratio and the overall Microcystis density. The highest proportions of toxic Microcystis genotypes were found 4–6 weeks before and 8–10 weeks after the peak of the bloom, with the lowest being observed at its peak. These results suggest positive selection of non-toxic genotypes under favorable environmental growth conditions. Significant positive correlations could be found between quantity of toxic genotypes and MC concentration, suggesting that the method applied can be useful to predict potential MC toxicity risk. No significant correlation was found between the limnological parameters measured and MC concentrations or toxic genotypes proportions indicating that other abiotic and biotic factors should be governing MC production and toxic genotypes dynamics. The qPCR method here applied is useful to rapidly estimate the potential toxicity of environmental samples and so, it may contribute to the more efficient management of water use in eutrophic systems.

The effects of aqueous extracts from Microcysts aeruginosa strains (both microcystin-producers and non-microcystin producers) on germination and root growth were investigated for three economically important plant species: Festuca rubra L., Lolium perenne L., and Lactuca sativa L. There was a clear inhibition of root growth for L. sativa exposed to strains containing microcystins (5.9-56.4 microg L(-1)). The strain that produced the most pronounced effects contained the lowest concentration of microcystin suggesting that other cellular compounds may also affect growth.

Mussels such as Mytilus galloprovincialis accumulate cyanotoxins without being affected by them. Microcystins (MCs), being the most common and studied cyanotoxins, covalently bind to protein phosphatases but this bond may be affected by environmental conditions. In this work, we studied the effect of different environmental and cooking conditions on the bioavailability of MCs in contaminated mussels. Conservation of contaminated mussels at room temperature (20 °C) led to a significant reduction of detectable MCs in the mussels. The conservation in the fridge (4 °C) produced a significant difference with a decrease on the detectable MC after 24, 48 and 72 h of exposure. The effects of the cooking on the MCs availability showed that boiling does not significantly change the MCs content of the mussels. Nevertheless, the microwave treatment significantly reduced the MCs content in both time periods used (1 and 5 min). Our work shows that the bioavailability of MCs in contaminated mussels may be changed due to conservation or cooking conditions.

Urban lakes are prone to eutrophication because of a number of factors, including a high water residence time (slow flushing rate), and high nutrient loads from artificial feeding of waterfowl and fish by visitors and from waterfowl faeces, falling leaves and nutrient run-off from adjacent areas. This study focuses on the ecology of two similar-sized urban lakes in Porto, Portugal, that exhibit some differing characteristics conducive to different trophic states and nutrient removal efficiencies. Although similar in many aspects, the two reservoirs developed quite diverse phytoplankton communities. The higher nitrogen load to Palacio de Cristal Lake could be the result of the higher concentration in its underground influent stream, while a less-oxidized nitrogen form (ammonia) is higher in Serralves Lake. Nitrate concentrations are higher in Palacio de Cristal Lake than in Serralves as a result mainly of high nitrate loads (maximum values above 80 mg L ‐1 ) in its influent stream. Interestingly, however, its nitrate removal efficiency is greater than for Serralves Lake. Cyanobacteria are the dominal phytoplankton In both lakes throughout the year. However, only one genus of this group (Pseudoanabaena mucicola) was the dominant phytoplankton in Palacio de Cristal Lake, while five Cyanobacteria species co-dominated in Serralves lake. Regular monitoring of Cyanobacteria and their toxins in urban recreation lakes is advised.

An increasing body of evidence points out that allelopathy may be an important process shaping microbial communities in aquatic ecosystems. Cyanobacteria have well-documented allelopathic properties, mainly derived from the evaluation of the activity of allelopathic extracts or pure compounds towards monocultures of selected target microorganisms. Consequently, little is known regarding the community dynamics of microorganisms associated with allelopathic interactions. In this laboratory-based study, a Microcystis spp.-dominated microbial community from a freshwater lake was exposed, for 15 days, to exudates from the cyanobacterium Oscillatoria sp. strain LEGE 05292 in laboratory conditions. This cyanobacterium is known to produce the allelochemicals portoamides, which were among the exuded compounds. The community composition was followed (by means of polymerase chain reaction followed by denaturing gradient gel electrophoresis and microscopic analyses) and compared to that of a non-exposed situation. Following exposure, clear differences in the community structure were observed, in particular for cyanobacteria and unicellular eukaryotic taxa. Interestingly, distinct Microcystis genotypes present in the community were differentially impacted by the exposure, highlighting the fine-scale dynamics elicited by the exudates. These results support a role for cyanobacterial allelochemicals in the structuring of aquatic microbial communities.

Cyanobacteria are prokaryotic photosynthetic microorganisms that pose a serious threat to aquatic environments because they are able to form blooms under eutrophic conditions and produce toxins. Cylindrospermopsis raciborskii is a planktonic heterocystous filamentous cyanobacterium initially assigned to the tropics but currently being found in more temperate regions such as Portugal, the southernmost record for this species in Europe. Cylindrospermopsin originally isolated from C. raciborskii is a cytotoxic alkaloid that affects the liver, kidney, and other organs. It has a great environmental impact associated with cattle mortality and human morbidity. Aiming in monitoring this cyanobacterium and its related toxin, a shallow pond located in the littoral center of Portugal, Vela Lake, used for agriculture and recreational purposes was monitored for a 2-year period. To accomplish this, we used the real-time PCR methodology in field samples to quantify the variation of specific genetic markers with primers previously described characterizing total cyanobacteria (16S rRNA), C. raciborskii (rpoC1), and cylindrospermopsin synthetase gene (pks). The results report the high abundance of both cyanobacteria and C. raciborskii in Vela Lake, with C. raciborskii representing 0.4% to 58% of the total cyanobacteria population. Cylindrospermopsin synthetase gene was detected in one of the samples. We believe that with the approach developed in this study, it will be possible to monitor C. raciborskii population dynamics and seasonal variation, as well as the potential toxin production in other aquatic environments.

A monitoring program of cyanobacteria and cyanotoxins in the framework of the surveillance of the Water Treatment Plant efficiency of the municipality of Santa Comba Dao (Portugal) was conducted from 1994 until 2007. With these data we aimed to answer the question, are MCs produced evenly over the years in a single water body? Samples were taken by the local health authorities in the site of the Water treatment plant and analyzed for total phytoplankton, cyanobacteria and the hepatotoxic cyanotoxins microcystins (MCs). Apart from 1999 and 2000, cyanobacteria represented a high proportion of total phytoplankton, attaining during several months of the year more than 90% of total phytoplankton density. A total of 24 cyanobacteria species were identified and Microcystis aeruginosa, Anabaena flos-aquae and Aphanizomenon flos-aquae were the main potentially toxic cyanobacteria species present throughout the period. MC concentration varied between 0.3 µg MC-LR eq l−1 in October 98 and 87.0 µg MC-LR eq l−1 in September 2001. The evolution of the average cell quota does not reveal any especial trend, although in 2001 the quota was the highest but not significantly different from the other years. The threshold limit of 5000 cells per ml of Microcystis aeruginosa should be taken into consideration in monitoring programs because the 10 000 cells per ml proposed by several other national programs might not be enough to prevent human health risks. Due to high annual variation in MC cell quota, monitoring programs of cyanobacteria and MC should be extended in time, since short term studies do not provide us the data needed for a safe management of a water body used for human purposes.

Although toxic cyanobacteria are commonly described worldwide, only one recent paper on the toxicity of cyanobacteria in Mexico has been published reporting the occurrence of cylindrospermopsin and saxitoxins. Microcystins are the most commonly studied cyanotoxins in many countries and those that may cause the most dramatic problems in terms of human health. In this paper, we studied the occurrence of potentially toxic cyanobacteria in different ecosystems in Central Mexico. Samples were collected in natural lakes (Zumpango, Laguna Atotonilco and Cienega Chica), reservoirs (Los Angeles and Valle de Bravo), man-made channels (Cuemanco, Tlameleca) and urban lakes (Chapultepec). A multi-technique approach was applied by the use of molecular, immunological and chemical techniques. Cyanobacteria were found in all the sites, ranging from 1.6 x 10(3) cells/mL in Tlameleca to 7.5 x 10(6) cells/mL in Chapultepec Grande, representing between 67 and 100% of the total phytoplankton density. The concentration of total microcystins varied between 4.9 and 78.0 microg MC-LR eq/L. The results revealed the occurrence of MC-LR in all the sites analyzed by MALDI-TOF, and MC-FR, MC-RR and MC-H(4)YR in two sites. Most of the studied sites are used either as a source of drinking water, as a recreational area, or for agriculture irrigation purposes, so the risk for human health may be high if not properly monitored. This is the first report on the MC profiles and concentrations in blooms collected in Mexico.

The aim of this work was to test the efficacy of molecular techniques for detecting toxigenic cyanobacteria in environmental water samples collected from freshwater lakes, rivers and reservoirs in Portugal. Of 26 environmental samples tested, 21 were found to contain Microcystis using a genus-specific polymerase chain reaction (PCR). Another primer pair was applied to the same DNA template to test for the presence of microcystin synthetase genes. This primer pair resulted in the formation of a PCR product in 15 of the samples containing Microcystis and one sample that did not give a positive result in the Microcystis genus-specific PCR. A restriction assay using the enzyme EcoRV was then applied to show that in most cases, the gene fragment was from toxigenic strains of Microcystis and, in one above-mentioned case, from a microcystin-producing strain of Planktothrix. All environmental samples were examined microscopically to confirm the presence of cyanobacteria species. Samples were also tested for the presence of microcystins using the ELISA plate assay. There was good agreement between the results obtained with molecular techniques and those obtained from microscopy and chemical methods. The PCR techniques applied in this paper were found to be useful, particularly when the concentration of the target organism was very low compared with other organisms. This technique can be used to detect inocula for cyanobacterial populations and therefore provide a useful tool for assessing under which conditions particular species can grow into bloom populations.