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Summary: The mean share of heterocystous cyanobacteria in total chlorophyll-a production in coastal waters, based on cyanobacterial marker carotenoid and chloropigments preserved in recent sediments (0–5 cm, ca 30 years), has been studied in the Gulf of Gdańsk (southern Baltic) and for comparison in the Oslofjord/Drammensfjord (southern Norway). First of all, Baltic cyanobacteria, both from laboratory cultures and field samples, were analysed to select marker heterocysteous cyanobacteria carotenoids for sediments. The pigment relation to diatom percentages of different salinity preferences has been tested, to confirm origin of cyanobacteria. The results indicate that canthaxanthin is the best marker of heterocystous cyanobacteria in the southern Baltic Sea. These filamentous cyanobacteria inflow to the Gulf of Gdańsk from the open sea and their abundance has increased in the last thirty years, in comparison with previous time. In that period they made up ca 4.6% of the total chlorophyll-a production in the Gulf of Gdańsk. The estimate for Oslofjord, at the same assumptions, suggests that heterocystous cyanobacteria occurred there also (up to 5.8% of the total chlorophyll-a production), were of marine origin, but their abundance has decreased during the last thirty years. Such an estimate may be used in environmental modelling and can be applied to other coastal areas, once the marker pigments of the main cyanobacteria species have been identified, and the percentage of total chlorophyll-a produced in a basin, preserved in sediments, has been determined for such area. Keywords: Cyanobacteria, Carotenoids, Canthaxanthin, Gulf of Gdańsk (southern Baltic), Oslofjord

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

Highlights • Fish mortalities due to harmful algae cause substantial economic and social costs for the fish farming industry in the northeastern Atlantic, North Sea and adjacent European waters • Toxin syndromes associated with Diarrhetic Shellfish Toxins and Paralytic Shellfish Toxins and their regulation have the most profound effect on the bivalve aquaculture industry in the northeastern Atlantic region • Cyanobacteria and cyanotoxins are mainly problems in brackish water areas, particularly in the Baltic Sea • Emerging threats to the shellfish and finfish industries include the known presence of the phycotoxins azaspiracids and goniodomins • The IOC-ICES-PICESHAEDAT contains time-series baseline information on harmful algal events in Europe Harmful algal blooms (HAB) are recurrent phenomena in northern Europe along the coasts of the Baltic Sea, Kattegat-Skagerrak, eastern North Sea, Norwegian Sea and the Barents Sea. These HABs have caused occasional massive losses for the aquaculture industry and have chronically affected socioeconomic interests in several ways. This status review gives an overview of historical HAB events and summarises reports to the Harmful Algae Event Database from 1986 to the end of year 2019 and observations made in long term monitoring programmes of potentially harmful phytoplankton and of phycotoxins in bivalve shellfish. Major HAB taxa causing fish mortalities in the region include blooms of the prymnesiophyte Chrysochromulina leadbeateri in northern Norway in 1991 and 2019, resulting in huge economic losses for fish farmers. A bloom of the prymesiophyte Prymnesium polylepis (syn. Chrysochromulina polylepis) in the Kattegat-Skagerrak in 1988 was ecosystem disruptive. Blooms of the prymnesiophyte Phaeocystis spp. have caused accumulations of foam on beaches in the southwestern North Sea and Wadden Sea coasts and shellfish mortality has been linked to their occurrence. Mortality of shellfish linked to HAB events has been observed in estuarine waters associated with influx of water from the southern North Sea. The first bloom of the dictyochophyte genus Pseudochattonella was observed in 1998, and since then such blooms have been observed in high cell densities in spring causing fish mortalities some years. Dinoflagellates, primarily Dinophysis spp., intermittently yield concentrations of Diarrhetic Shellfish Toxins (DST) in blue mussels, Mytilus edulis, above regulatory limits along the coasts of Norway, Denmark and the Swedish west coast. On average, DST levels in shellfish have decreased along the Swedish and Norwegian Skagerrak coasts since approximately 2006, coinciding with a decrease in the cell abundance of D. acuta. Among dinoflagellates, Alexandrium species are the major source of Paralytic Shellfish Toxins (PST) in the region. PST concentrations above regulatory levels were rare in the Skagerrak-Kattegat during the three decadal review period, but frequent and often abundant findings of Alexandrium resting cysts in surface sediments indicate a high potential risk for blooms. PST levels often above regulatory limits along the west coast of Norway are associated with A. catenella (ribotype Group 1) as the main toxin producer. Other Alexandrium species, such as A. ostenfeldii and A. minutum, are capable of producing PST among some populations but are usually not associated with PSP events in the region. The cell abundance of A. pseudogonyaulax, a producer of the ichthyotoxin goniodomin (GD), has increased in the Skagerrak-Kattegat since 2010, and may constitute an emerging threat. The dinoflagellate Azadinium spp. have been unequivocally linked to the presence of azaspiracid toxins (AZT) responsible for Azaspiracid Shellfish Poisoning (AZP) in northern Europe. These toxins were detected in bivalve shellfish at concentrations above regulatory limits for the first time in Norway in blue mussels in 2005 and in Sweden in blue mussels and oysters (Ostrea edulis and Crassostrea gigas) in 2018. Certain members of the diatom genus Pseudo-nitzschia produce the neurotoxin domoic acid and analogs known as Amnesic Shellfish Toxins (AST). Blooms of Pseudo-nitzschia were common in the North Sea and the Skagerrak-Kattegat, but levels of AST in bivalve shellfish were rarely above regulatory limits during the review period. Summer cyanobacteria blooms in the Baltic Sea are a concern mainly for tourism by causing massive fouling of bathing water and beaches. Some of the cyanobacteria produce toxins, e.g. Nodularia spumigena, producer of nodularin, which may be a human health problem and cause occasional dog mortalities. Coastal and shelf sea regions in northern Europe provide a key supply of seafood, socioeconomic well-being and ecosystem services. Increasing anthropogenic influence and climate change create environmental stressors causing shifts in the biogeography and intensity of HABs. Continued monitoring of HAB and phycotoxins and the operation of historical databases such as HAEDAT provide not only an ongoing status report but also provide a way to interpret causes and mechanisms of HABs.

Mercury is a toxic element. It undergoes biomagnification in the marine trophic chain, which is why it is significant to identify the factors influencing its bioaccumulation on the first level of the trophic chain. At present, the input of heavy metals to the southern Baltic is being reduced. On the other hand, the parameters influencing mercury remobilisation in the environment are a subject to a long-time trend associated with climate changes. Examples include growing number of heavy rain events causing surges or floods, and increased frequency of storm winds leading to increased coastal erosion as well as overall temperature increase. The present studies were carried out in the coastal zone of the Gulf of Gdańsk (southern Baltic) for 18 months at two stations (Chałupy and Osłonino) located in the Puck Lagoon, and for 12 months in Gdynia. Climate changes influence the abundance and species composition of phytoplankton, which in consequence has an effect on Hg accumulation and magnification in the trophic chain, and in the human body as a result. Extreme phenomena such as land erosion or floods resulted in an additional inflow of nutrients, but also toxic substances, into the coastal zone. The bioconcentration factor (BCF) increased almost four times after abrasion of cliff. That was conducive to the growth of microflora, as well as increased Hg accumulation. The highest bioconcentration of Hg in phytoplankton was observed when the Mesodinium rubrum (spring and autumn) and Diatomophyceae (winter) prevailed in biomass. The BCF was then almost tenfold higher than during the rest of the year.

This study aims to demonstrate that factors associated with climate dynamics, such as temperature and wind, affect the ecosystem of the shallow Vistula Lagoon in the southern Baltic and cause nutrient forms phytoplankton interactions: the growth of biomass and constraints of it. This occurs through a network of direct and indirect relationships between environmental and phytoplankton factors, including interactions of positive and negative feedback loops. Path analysis supported by structural equation modeling (SEM) was used to test hypotheses regarding the impact of climate factors on algal assemblages. Increased phytoplankton biomass was affected directly by water temperature and salinity, while the wind speed effect was indirect as it resulted in increased concentrations of suspended solids (SS) in the water column. Simultaneously, the concentration of SS in the water was positively correlated with particulate organic carbon (POC), particulate nitrogen (PN), and particulate phosphorus (PP), and was negatively correlated with the total nitrogen to phosphorus (N:P) ratio. Particulate forms of C, N, and phosphorus (P), concentrations of soluble reactive phosphorus (SRP) and nitrate and nitrite nitrogen (NO3-N + NO2-N), and ratios of the total N:P and DIN:SRP, all indirectly effected Cyanobacteria C concentrations. These processes influence other phytoplankton groups (Chlorophyta, Bacillariophyceae and the picophytoplankton fraction). Increased levels of SRP associated with organic matter (POC), which stemmed from reduced DIN:SRP ratios, contributed to increased Cyanoprokaryota and picophytoplankton C concentrations, which created a positive feedback loop. However, a simultaneous reduction in the total N:P ratio could have inhibited increases in the biomass of these assemblages by limiting N, which likely formed a negative feedback loop. The study indicates that the nutrients-phytoplankton feedback loop phenomenon can intensify eutrophication in a temperate lagoon, including increases of the biomass of Cyanobacteria and picophytoplankton. However, it can also constrain this increase.

Studies of cyanobacterial bloom dynamics show that the highest biomass accumulation of Nodularia spumigena is observed in the shallowest area of the Gulf of Gdańsk in summer. In the same region and time, the highest fish abundance is observed. Mostly young individuals of gobies, small sandeel, flounder, three-spine stickleback and young herring occur. In this work we compare how toxic blooms of cyanobacteria influence the number and structure of fish communities in a coastal zone. The results obtained in our study were rather unexpected. More fish species were caught and the biomass of fish was higher during a bloom than in a month following the sampling (no bloom).

Almost every summer, dense blooms of filamentous cyanobacteria are formed in the Baltic Sea. These blooms may cause problems for tourism and ecosystem services, where surface accumulations and beach fouling are commonly occurring. Future changes in environmental drivers, including climate change and other anthropogenic disturbances, may further enhance these problems. By compiling monitoring data from countries adjacent to the Baltic Sea, we present spatial and temporal genus-specific distribution of diazotrophic filamentous cyanobacteria (Nostocales) during four decades (1979–2017). While the summer surface salinity decreased with a half up to one unit, the surface temperature in July-August increased with 2–3 °C in most sub-basins of the Baltic Sea, during the time period. The biovolumes of the toxic Nodularia spumigena did not change in any of the sub-basins during the period. On the other hand, the biovolume of the non-toxic Aphanizomenon sp. and the potentially toxic Dolichospermum spp. increased in the northern parts of the Baltic Sea, along with the decreased salinity and elevated temperatures, but Aphanizomenon sp. decreased in the southern parts despite decreased salinity and increased temperatures. These contradictory changes in biovolume of Aphanizomenon sp. between the northern and southern parts of the Baltic Sea may be due to basin-specific effects of the changed environmental conditions, or can be related to local adaptation by sub-populations of the genera. Overall, this comprehensive dataset presents insights to genus-specific bloom dynamics by potentially harmful diazotrophic filamentous cyanobacteria in the Baltic Sea. Highlights • Biovolumes of bloom-forming cyanobacteria during four decades in the Baltic Sea. • Aphanizomenon sp. has increased with decreased salinity in the Bothnian Sea. • Dolichospermum spp. has increased with temperature in Bothnian Sea. • The total biovolume of Nostocales has decreased in the Southern Baltic Proper. • The biovolume of the toxic Nodularia spumigena has not changed since the 1980s.

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.

Mercury is a neurotoxin, its main source in the human organism being fish and seafood. The first level in the marine food web is formed of planktonic and benthic photosynthetic microorganisms, which form a biofilm on the surface of the hard bottom (epilithon) or plants (epiphyton). They are carriers of nutritional as well as toxic substances and pass these on to subsequent levels of the trophic web. Their biomass is often dominated by diatoms. This was the basis for the presented study into Hg accumulation in epilithic, epiphytic and planktonic diatoms, which was carried out in 2012–2013 in the coastal zone of the Puck Lagoon and the Gulf of Gdańsk (southern Baltic). In this coastal area, both micro- and macroorganisms develop particularly intensively. The collected results indicate an increase in Hg concentration in the biofilm during the warm season which, with the lengthening of the vegetative period due to global warming in recent years, is of great significance. As a consequence, the annual mercury load entering the trophic web is larger in comparison with a year in which there is a long, cold winter. An important parameter influencing the accumulation of Hg was the function of those organisms from the biofilm-forming communities. In this case, the highest concentrations of Hg were measured in organisms forming high-profile guilds. Electronic supplementary material The online version of this article (10.1007/s11270-018-3814-1) contains supplementary material, which is available to authorized users.

The threat connected to mercury results from its capacity to be transported over long distances and its ability to bioaccumulate and biomagnify in the trophic chain, making it a global problem. Humans are situated at the top of the trophic ladder, and excess mercury manifests itself mainly in the onset of neurological conditions. The toxicity of mercury, as well as its residence time, depends on the form in which it occurs. However, analysis of mercury speciation is time-consuming and poses a high risk of additional or negative contamination. Hence, the mercury thermodesorption method, and particularly its use for fractionating Hg, offers many new possibilities. Here, the thermodesorption technique was applied to the determination of mercury fraction in particulate matter using a DMA-80 direct mercury analyser (Milestone, Italy). The presented method allows direct (without prior mineralisation) determination of labile and stabile mercury fractions within a relatively short time. Heating sample in subsequent temperatures enables determination the share of mercury adsorbed on the surface (mainly associated with halogenides (Hgads1) and HgSO4/HgO/HgF2 (Hgads2), as well as absorbed within the suspended particulate organic matter (Hgabs), in a relatively short time. This fractionation is of great importance in terms of estimating the transfer of mercury to and along the trophic chain. This method determines the contribution of two stable mercury fractions:: HgS and residual Hg, strongly bound to particulate matter matrix (Hgres). The novelty of this technique is also its joint ability to determine gaseous mercury bound to airboirne particulate matter, which will enable better understand Hg cycling in the atmosphere as well as mercury fraction in dry deposition flux. This method enables assessment of global mercury circulation in environment.

Mechanisms behind expansion of an invasive cyanobacterium Cylindrospermopsis raciborskii have not been fully resolved, and different hypotheses, such as global warming, are suggested. In the East-Central Europe, it is widely occurring in western part of Poland but only in single locations in the East due to some limiting factors. Therefore, broad-scale phytoplankton survey including 117 randomly selected lakes in Poland and Lithuania was conducted. The results showed that C. raciborskii occurred widely in western part of Poland but was absent from other regions and Lithuania except one lake. The regions in which C. raciborskii was present had higher annual mean air temperature, higher maximum air temperature of the warmest month and higher minimum temperature of the coldest month, demonstrating that average air temperature, and indirectly, the duration of growing season might be more important factor driving C. raciborskii distribution than measured in situ water temperature. In turn, the presence of C. raciborskii in single localities may be more related to physiological adaptations of separated ecotype. Collectively, these results provide novel evidence on the influence of temperature on C. raciborskii distribution in East-European regions but also indicate high ecological plasticity of this species.

Planktothtrix agardhii (Oscillatoriales) is a filamentous cyanobacterium, which frequently forms blooms in shallow, polymictic and eutrophicated waters. This species is also a rich source of unique linear and cyclic peptides. In the current study, the profile of the peptides in samples from the P. agardhii-dominated Siemianówka Dam Reservoir (SDR) (northeast Poland) was analyzed for four subsequent years (2009–2012). The LC–MS/MS analyses revealed the presence of 33 peptides. Twelve of the most abundant ones, including five microcystins, five anabaenopeptins, one aeruginosin and one planktocyclin, were present in all field samples collected during the study. The detection of different peptides in two P. agardhii isolates indicated that the SDR population was composed of several chemotypes, characterized by different peptide patterns. The total concentration of microcystins (MCs) positively correlated with the biomass of P. agardhii. Between subsequent years, the changes in the ratio of the total MCs concentration to the biomass of P. agardhii were noticed, but they were less than threefold. This is the first study on the production of different classes of non-ribosomal peptides by freshwater cyanobacteria in Poland. Electronic supplementary material The online version of this article (doi:10.1007/s00203-014-1008-9) contains supplementary material, which is available to authorized users.

The biovolume of phytoplankton must be assessed accurately in order to identify the ecological status of water bodies in line with the WFD requirements. Hence, the current study has been carried out to verify and improve the precision of as well as to facilitate and accelerate estimations of phytoplankton biovolume by reviewing and rearranging the basic geometrical shapes of these organisms applied in such evaluations. The latest standards comprise 17 geometric shapes and equations suitable for estimations of cell/filament/colony biovolume and additionally include taxa-specific ‘geometric correction factors’ to fit real shapes and ‘hidden dimension factors’ to achieve data on hardly measureable dimensions. This paper also discusses possible obstacles to making correct biovolume assessments, especially when analyzing taxa of special concern, e.g. Ceratium hirundinella, C. furcoides or Pediastrum duplex and Pseudopediastrum boryanum. Our comparison of two approaches, the previous and the new one, revealed that they yield statistically significantly different biovolume results of these species. Some recommendations how to deal with the new and old methods of biovolume estimations and how to reduce the possibility of errors with overestimation and underestimation were also given. The more recent method can be said to give more precise estimates of phytoplankton biovolume. Besides, it facilitates more rapid phytoplankton analyses in most cases, which is very useful when assessing the ecological status of lakes during routine monitoring programs.

The Vistula Lagoon (southern Baltic Sea) is a shallow and highly eutrophic water body, with frequent blooms of cyanobacteria dominated by Microcystis and Anabaena species. Two Microcystis strains, MK10.10 and MAKR0205, isolated from the lagoon were characterised in this work. The morphology of the isolates differed significantly with respect to cell size and their ability to form aggregates.Based on the 16S rRNA sequence and 16S-23S internal transcribed spacer (ITS) sequence, both isolates were classified as Microcystis aeruginosa. However, only one isolate, MK10.10, possessed the mcygenes responsible for microcystin biosynthesis and only this strain produced microcystins. The effects of environmental factors, such as light, temperature and salinity, on toxin production turned out to be minor. Under the culture conditions used in the experiments, the biomass of the toxic MK10.10 was alwayslower. Hybrid quadrupole-time-of-flight liquid chromatography/tandem mass spectrometry (QTOF-LC/MS/MS) was used to elucidate the structure of the microcystin (MC) variants produced by MK10.10. Based on molecular ion and fragmentation spectra, the toxins were identified as MC-LR, MC-VR and MC-HIlR. Our study confirmed that some morphological criteria could be useful in preliminarily assessing the potential toxicity of a Microcystis bloom.

Microcystins (MCs) and structurally related nodularins (NODs) are hepatotoxic cyclic peptides produced by bloom-forming cyanobacteria. These toxins have been implicated in the deaths of wild and domestic animals as well as in incidents of human illness. Cyanobacterial toxins occurring in the fresh and brackish waters of the Pomeranian Province, northern Poland were characterized in this study. Water samples collected from seven lakes in August and September 2005 were analysed by high performance liquid chromatography (HPLC), enzyme linked immunosorbent assay (ELISA) and protein phosphatase inhibition assay (PPIA). Cyanobacterial toxins present in field samples and in an isolated strain of Planktothrix agardhii were also characterized by liquid chromatography electrospray ionization tandem mass spectrometry (LC-ESI-MS/MS). In most of the fresh water samples MC-LR, MC-RR and MC-YR dominated. In the lakes where P. agardhii was most abundant demethylated microcystin variants tentatively identified as [D-Asp3]MC-LR, [D-Asp3]MC-YR and [D-Asp3]MC-RR, were found. Total concentrations of the toxins measured by HPLC ranged from 0.1 μg 1-1 to 305.4 μg 1-1. Nodularia spumigena bloom samples were collected from brackish waters of the Gulf of Gdańsk, southern Baltic, and LC-ISP-MS/MS of extract from these revealed the presence of two geometrical isomers of linear nodularin and nodularin variant with aspartic acid methyl ester [MeAsp1(OMe)]NOD.

Nodularia spumigena Mertens, Aphanizomenon flos-aquae (L.) Ralfs and some species of the genus Anabaena are the dominant cyanobacterial taxa occurring in the Gulf of Gdańsk. The heterocystous cyanobacteria use dissolved molecular N2 as an additional nitrogen source, and this allows them to bloom during the summer when growth of other phytoplankton species is normally nitrogen-limited. Although cyanobacterial blooms have been reported in the Baltic Sea since the mid-19th century, the extent and intensity of blooms have recently increased due to anthropogenic sources of eutrophication. Increased river phosphorus input and changes in the phosphorus to nitrogen ratio are implicated as causal factors. After us the initial cause of the cyanobacterial bloom is a low N:P ratio, which indicates phosphorus excess, i.e. favourable nutrient conditions for nitrogen-fixing algae. An N:P ratio of 10 has been considered an approximate value for the N:P requirements of Baltic phytoplankton. For several years this ratio has been lower than 10. The mean annual value of the N:P ratio for the water of the Gulf of Gdańsk ranged from 3 to 7. Differences in the intensity of blooms observed in different years could be linked to temperature. During hot summers, when the seawater temperature increased to 20°C, large blooms were noted. For the cyanobacterial blooms in the Baltic Sea, the low N:P ratio is the primary factor and high temperature is a starting point.

Cyanobacteria are effective producers of bioactive metabolites, including both acute toxins and potential pharmaceuticals. In the current work, the biological activity of 27 strains of Baltic cyanobacteria representing different taxonomic groups and chemotypes were tested in a wide variety of assays. The cyanobacteria showed strain-specific differences in the induced effects. The extracts from Nodularia spumigena CCNP1401 were active in the highest number of tests, including protease and phosphatase inhibition assays. Four strains from Nostocales and four from Oscillatoriales increased proliferation of mitogen-stimulated human T cells. In antimicrobial assays, Phormidium sp. CCNP1317 (Oscillatoriales) strongly inhibited the growth of six fouling Gammaproteobacteria. The growth of monocotyl Sorghum saccharatum was inhibited by both toxin-producing and ‘non-toxic’ strains. The Baltic cyanobacteria were also found to be a rich source of commercially important enzymes. Among the 19 enzymatic activities tested, alkaline phosphatase, acid phosphatase, esterase (C4 and C8), and naphthol-AS-BI-phosphohydrolase were particularly common. In the cyanobacterial extracts, different peptides which may have been responsible for the observed effects were identified using LC-MS/MS. Their structures were classified to microcystins, nodularins, anabaenopeptins, cyanopeptolins, aeruginosins, spumigins and nostocyclopeptides.

Background: It is generally believed that there are many natural sources of as yet unknown bioactive compounds with a high biotechnological potential. However, the common method based on the use of cell extracts in the preliminary screening for particular molecules or activities is problematic as amounts of obtained compounds may be low, and such experiments are hardly reproducible. Therefore, the aim of this work was to test whether a novel strategy to search for previously unknown biological activities can be efficient. This strategy is based on construction of metagenomic libraries and employment of Escherichia coli strains as cell factories producing compounds of properties potentially useful in biotechnology. Results: Three cyanobacterial metagenomic libraries were constructed in the fosmid system. The libraries were screened for various biological activities. Extracts from selected E. coli clones bearing constructs with fragments of cyanobacterial genomes revealed antimicrobial or anticancer activities. Interestingly, stimulation of growth of host bacteria bearing particular plasmids with certain cyanobacterial genes was detected, suggesting a potential possibility for improvement of E. coli cultivation during biotechnological production. The most interesting plasmids were sequenced, and putative mechanisms of biological effects caused by cyanobacterial gene products are discussed. Conclusions: The strategy of exploring cyanobacteria as sources of bioactive compounds, based on E. coli cell factories producing compounds due to expression of genes from metagenomic libraries, appears to be effective.

In this work, the authors examined the presence of cyanobacteria and cyanotoxins in 21 samples collected from fresh water bodies located in 5 provinces in Poland: Lublin (2), Podlasie (1), Pomerania (6), Warmia-Masuria (1) and Wielkopolska (11). In addition, to determine the general pattern of geographical distribution, frequency of cyanobacteria occurrence, and cyanotoxins production, the published data from 238 fresh water bodies in Poland were reviewed. On the basis of these collected results, we concluded that Planktothrix, Aphanizomenon, Microcystis and Dolichospermum were dominant. The general pattern in geographical distribution of the identified cyanobacterial genera was typical of other eutrophic waters in Europe. The production of cyanotoxins was revealed in 18 (86%) of the 21 samples analyzed in the present work and in 74 (75%) of the 98 total water bodies for which the presence of toxins had been examined. Among the 24 detected microcystin variants, [Asp3]MC-RR was most common. These results can be verified when more data from the less explored water bodies in the southern and eastern parts of Poland are available. The authors would like to acknowledge the European Cooperation in Science and Technology, COST Action ES 1105 "CYANOCOST- Cyanobacterial blooms and toxins in water resources: Occurrence, impacts and management" for adding value to this study through networking and knowledge sharing with European experts and researchers in the field. 42 4 358 378

In the Baltic Sea, cyanobacterial community is mainly composed of filamentous nitrogen-fixing forms, including the toxic Nodularia spumigena, and single-celled picocyanobacteria (Pcy), represented by Synechococcus spp. The main aim of the work was to test the hypothesis that the picocyanobacteria dependend on the presence of the nitrogen-fixing cyanobacteria. In addition, the contamination of blue mussels and fish with nodularin (NOD), the N. spumigena toxin, was examined. In years 2008–2011, the samples for the study were collected in the Southern Baltic Proper using FerryBox system and, occasionally, during research cruises. The analyses showed no correlation between the growth of the nitrogen-fixing cyanobacteria and Synechococcus. Compared with the previously published data, a shift in the composition of Pcy phenotypes was observed. This shift might be an indication of the proceeding changes induced by the reduced nutrient loading and/or climate change. Analyses of NOD revealed differences in the cyanotoxin concentrations between mussels of different shell size. The highest concentration of NOD was detected in the liver of round goby. However, temporarily, also the fish muscles were significantly contaminated with the toxin.