A Fuzzy Logic Model to Describe the Cyanobacteria Nodularia spumigena Blooms in the Gulf of Finland, Baltic Sea

A fuzzy logic model to describe the seasonal evolution of Nodularia spumigena blooms in the Gulf of Finland was built and calibrated on the basis of monitoring data. The model includes three phosphate sources: excess phosphate after the annual spring bloom and parameterised phosphate transport to the upper mixed layer by turbulent mixing and upwelling events. Surface layer temperature and wind mixing form the physical conditions controlling the growth of N. spumigena. Model simulations revealed that phosphate input caused by turbulent mixing and upwelling have to be taken into account to achieve the best fit with observed data. Testing the fuzzy model for early prediction of maximum N. spumigena biomass about a month before the usual occurrence of blooms, gave good results. The potential use of the model for prediction of bloom risk at a certain location along the Estonian or Finnish coast was tested. The bloom transport velocities used in the fuzzy model were pre-calculated by a 3D numerical circulation model for different wind regimes.     

Effect of salinity on photosynthetic activity of Nodularia spumigena

The aim of the study was to determine the influence of total dissolvedsolids/salinity (mg L^-1 TDS) on photosynthetic activity of Nodularia spumigena strain 001E isolated from Lake Alexandrina, SouthAustralia, using photosynthesis-irradiance (PI) curves. N. spumigena001E cultures were grown in ASM medium at a range of TDSconcentrations (360, 6,600, 13,200, 19,800, 26,400 mg L^-1)at an irradiance of 30 mol m^-2 s^-1 (PAR, 400–700 nm) at 25 °C. The PI relationship was determined at 25 °Cfor irradiances between 0 and 500 mol photon m^-2s^-1 (PAR). The initial slope of PI curve, , a function of lightharvesting efficiency and photosynthetic energy conversion, decreasedproportionally with an increase in salinity from 360 to 26,400 mgL^-1 TDS. The maximum rate of photosynthesis (P_max),occurred at 6,600 mg L^-1 TDS. No influence of salinity onI_k, the irradiance at which P_max was measured, or on R_d, the dark respiration rate, was identified.     

Effect of irradiance, temperature and salinity on growth and toxin production by Nodularia spumigena

The object of this work was to determine, using a full-factorial experiment, the influence of temperature, irradiance and salinity on growth and hepatotoxin production by Nodularia spumigena, isolated from Lake Alexandrina in the south-east of South Australia. Higher levels of biomass (determined as particulate organic carbon, POC), toxin production and intracellular toxin concentration per mg POC were produced under light limited conditions (30 mol m^–2 s^–1) and at salinities equal to or greater than those experienced in Lake Alexandrina. Both highest biomass and total toxin production rates were recorded at temperatures equal to or greater than those of the lake (20 and 30°C). The temperature at which maximum biomass and toxin production was recorded decreased from 30°C for cultures grown at 30 mol m^–2 s^–1 to 20°C when grown at 80 mol m^–2 s^–1. In contrast, intracellular toxin per mg POC was highest at the lowest growth temperature, 10°C, at both 30 and 80 mol m^–2 s^–1. It appears that the optimum temperature for biosynthetic pathways used in the production of toxin is lower than the optimum temperature for those pathways associated with growth. Intracellular toxin levels were higher in cells cultured at 10°C/30 mol m^–2 s^–1 whereas the majority of the toxin was extracellular in cells grown at 30°C/30 mol m^–2 s^–1. This implies that the highest concentration of toxin in lake water would occur under high temperature and high irradiance conditions. Individual environmental parameters of salinity, irradiance and temperature were all shown to influence growth and toxin production. Notwithstanding, the overall influence of these three parameters on toxin production was mediated through their effect upon growth rate.     

Sedimentation of Nodularia spumigena and distribution of nodularin in the food web during transport of a cyanobacterial bloom from the Baltic Sea to the Kattegat

Nodularia spumigena is a toxic cyanobacteria that blooms in the Baltic Sea every year. In the brackish water of the Baltic Sea, its toxin, nodularin, mainly affects the biota in the surface water due to the natural buoyancy of this species. However, the fate of the toxin is unknown, once the cyanobacteria bloom enters the more saline waters of the Kattegat. In order to investigate this knowledge gap, a bloom of N. spumigena was followed during its passage, carried by surface currents, from the Baltic Sea into the Kattegat area, through the Öresund strait. N. spumigena cells showed an increased cell concentration through the water column during the passage of the bloom (up to 130 10³ cells ml⁻¹), and cells (4.2 10³ cells ml⁻¹) could be found down to 20 m depth, below a pycnocline. Sedimentation trap samples from below the pycnocline (10–12 m depth) also showed an increased sedimentation of N. spumigena filaments during the passage of the bloom. The toxin nodularin was detected both in water samples (0.3–6.0 μg l⁻¹), samples of sedimenting material (a toxin accumulation rate of 20 μg m^-2 day⁻¹), zooplankton (up to 0.1 ng ind.⁻¹ in copepods), blue mussels (70–230 μg kg⁻¹ DW), pelagic and benthic fish (herring (1.0–3.4 μg kg⁻¹ DW in herring muscle or liver) and flounder (1.3-6.2 μg kg⁻¹ DW in muscle, and 11.7-26.3 μg kg⁻¹ DW in liver). A laboratory experiment showed that N. spumigena filaments developed a decreased buoyancy at increased salinities and that they were even sinking with a rate of up to 1,7 m day⁻¹ at the highest salinity (32 PSU). This has implications for the fate of brackish water cyanobacterial blooms, when these reach more saline waters. It can be speculated that a significant part of the blooms content of nodularin will reach benthic organisms in this situation, compared to blooms decaying in brackish water, where most of the bloom is considered to be decomposed in the surface waters.     

Intraspecific variation in the response of the cyanobacterium Nodularia spumigena to moderate UV-B radiation

Floating and nodularin-producing strains of Nodularia spumigena from the Baltic Sea are regarded as belonging to one species. However, intraspecific variation in the response of N. spumigena to environmental factors has been commonly overlooked. As blooms of N. spumigena occur in late summer, a period with strong light and stable water-column stratification, the cells can be expected to also be exposed to ultraviolet-B radiation (UV-B, 280–320 nm). The UV-B tolerance of four different strains of N. spumigena, isolated from the Baltic Sea, was investigated in the laboratory for 8 days, by measuring photosynthesis, growth and pigment composition. Variables included maximum quantum yield of photosynthesis (F_v/F_m, PAM fluorometry), growth rate (cell counts) and photosynthetic pigments, as well as mycosporine-like amino acids (HPLC). Intraspecific differences regardless of treatment were found for cell dimension, growth rate, F_v/F_m and pigment concentrations. UV-B related effects differed between strains. By Day 8 one of the four strains showed a lower F_v/F_m when treated with UV-B; in another strain the growth rate and cell numbers were lower. In three strains, UV-B exposure resulted in higher cell concentrations of carotenoids and chlorophyll a. In all strains, the concentrations of total mycosporine-like amino acids were 60–130% higher in the UV-B treated samples compared with samples shielded from UV-B. Although strain-specific differences in UV-B tolerance were observed, it is concluded that N. spumigena is a species that is not generally negatively affected by moderate levels of UV-B radiation.     

Effects of Cyanobacteria on Survival and Reproduction of the Littoral Crustacean Gammarus zaddachi (Amphipoda)

Toxic cyanobacteria can have harmful or fatal impacts on aquatic organisms. In the archipelagos of the northern Baltic Sea, the open sea blooms often drift into littoral areas, where they decompose and release toxins and other chemical compounds in the water. However, the effects of cyanobacteria on the littoral organisms have not previously been investigated. We studied the effects of three cyanobacteria species (toxic Nodularia spumigena, non-toxic N. sphaerocarpa and non-toxic Aphanizomenon flos-aquae) and purified dissolved nodularin (produced by N. spumigena) on a common littoral amphipod Gammarus zaddachi. Nodularin was transferred to eggs, juveniles and adults of G. zaddachi, but no significant negative effects of dissolved nodularin were detected on adults, eggs or juveniles. However, survival of adults decreased by the exposure to toxic N. spumigena cells. The egg hatching rate and juvenile survival were not affected when exposed to the three cyanobacteria species. In contrast, a weak decrease in the egg production and an increased abortion of embryos from the brood pouch of females was observed, the later indicating a failure in parental care. Further, a decrease in grazing rate on the filamentous green alga Enteromorpha intestinalis was observed. The results suggest that toxic cyanobacteria blooms are not extremely fatal, but may have, in high concentrations, negative effects on the adult survival, fecundity, and feeding behaviour of gammarids inhabiting the littoral zone.     

Nitrogen Fixation During Blooms of Nodularia in Coastal Waters and Backwaters of the Arkona Sea (Baltic Sea) in 1974

Nitrogen fixation was investigated by means of the acetylene reduction method during the development of a water bloom of Nodularia in coastal waters of the Baltic Sea west of the island of Hiddensee and in backwaters showing different degrees of eutrophication. Depending on plankton density, the values found varied greatly. The maximum of nitrogen fixation values found in extremely dense water blooms under special conditions (Baltic Sea, 2250 μg N₂/l · h; Kleiner Jasmunder Bodden, 374 μg N₂/l · h) are up to 10³ times higher than from other parts of the Baltic Sea or from inland waters. The average nitrogenase activity determined for coastal water populations of the Baltic Sea is 2.15 pg N₂/heterocyst · h and that of the inmost backwaters 0.77 pg N₂/heterocyst · h. The relationship between N₂-fixation and nutrient content in water is discussed.     

Carbon,nitrogen and O2 fluxes associated with the cyanobacterium Nodularia spumigena in the Baltic Sea

Photosynthesis, respiration, N₂ fixation and ammonium release were studied directly in Nodularia spumigena during a bloom in the Baltic Sea using a combination of microsensors, stable isotope tracer experiments combined with nanoscale secondary ion mass spectrometry (nanoSIMS) and fluorometry. Cell-specific net C- and N₂-fixation rates by N. spumigena were 81.6±6.7 and 11.4±0.9 fmol N per cell per h, respectively. During light, the net C:N fixation ratio was 8.0±0.8. During darkness, carbon fixation was not detectable, but N₂ fixation was 5.4±0.4 fmol N per cell per h. Net photosynthesis varied between 0.34 and 250 nmol O₂ h⁻¹ in colonies with diameters ranging between 0.13 and 5.0 mm, and it reached the theoretical upper limit set by diffusion of dissolved inorganic carbon to colonies (>1 mm). Dark respiration of the same colonies varied between 0.038 and 87 nmol O₂ h⁻¹, and it reached the limit set by O₂ diffusion from the surrounding water to colonies (>1 mm). N₂ fixation associated with N. spumigena colonies (>1 mm) comprised on average 18% of the total N₂ fixation in the bulk water. Net NH₄⁺ release in colonies equaled 8–33% of the estimated gross N₂ fixation during photosynthesis. NH₄⁺ concentrations within light-exposed colonies, modeled from measured net NH₄⁺ release rates, were 60-fold higher than that of the bulk. Hence, N. spumigena colonies comprise highly productive microenvironments and an attractive NH₄⁺ microenvironment to be utilized by other (micro)organisms in the Baltic Sea where dissolved inorganic nitrogen is limiting growth.     

Occurrence of the hepatotoxic cyanobacterium Nodularia spumigena in the Baltic Sea and structure of the toxin

Water blooms formed by potentially toxic species of cyanobacteria are a common phenomenon in the Baltic Sea in late summer. Twenty-five cyanobacterial bloom samples were collected from open and coastal waters of the Baltic Sea during 1985 to 1987, and their toxicity was determined by mouse bioassay. All of 5 bloom samples from the southern Baltic Sea, 6 of 6 from the open northern Baltic Sea (Gulf of Finland), and 7 of 14 Finnish coastal samples were found to contain hepatotoxic cyanobacteria. Nodularia spumigena and Aphanizomenon flos-aquae occurred together in high amounts in blooms from the open-sea areas. In addition, coastal samples contained the species Anabaena lemmermannii, Microcystis aeruginosa, and Oscillatoria agardhii. Eighteen hepatotoxic N. spumigena cultures were isolated from water bloom and open-sea water samples. High-pressure liquid chromatographic analysis of both hepatotoxic bloom samples and Nodularia strains showed a single toxic fraction. The toxin concentrations of the blooms were less than or equal to 2.4 mg/g of freeze-dried material, and those of laboratory-grown cultures were 2.5 to 8.0 mg/g of freeze-dried cells. A single toxin was isolated from three N. spumigena-containing bloom samples and three N. spumigena laboratory isolates. Amino acid analysis and low- and high-resolution fast-atom bombardment mass spectroscopy indicated that the toxin from all of the sources was a cyclic pentapeptide (molecular weight, 824) containing glutamic acid, beta-methylaspartic acid, arginine, N-methyldehydrobutyrine, and 3-amino-9-methoxy-2,6,8-trimethyl-10-phenyl-4,6-decadienoic acid.(ABSTRACT TRUNCATED AT 250 WORDS)     

Estimating nodularin content of cyanobacterial blooms from abundance of Nodularia spumigena and its characteristic pigments—a case study from the Baltic entrance area

The presence of toxin-producing cyanobacteria is of concern for the management of recreational waters. The abundance of toxic cyanobacteria and environmental concentrations of their toxins may fluctuate substantially within a short time emphasising the need for rapid methods to estimate toxin concentrations in the water. During late June-early September 2002 the abundance, biomass and characteristic pigments of Nodularia spumigena Mertens and the hepatotoxin nodularin produced by N. spumigena were analysed in water samples collected from the Baltic entrance area. Significant relationships were found between cell-bound concentrations of nodularin and the abundance and biomass of N. spumigena with a relationship of approximately 1 pg nodularin per Nodularia-cell. It is suggested that simple counts of Nodularia under the microscope may be used as a rapid on-site technique to estimate potential nodularin concentrations in recreational waters. Comparison with data from Australia shows that cell-bound concentration of nodularin per Nodularia-cell differs between geographically distant areas and therefore such relationships should be established for individual areas. The carotenoids echinenone, canthaxanthin and a cis-canthaxanthin-like caroteniod, identified from a laboratory culture of N. spumigena isolated from the Baltic Sea, were also significantly correlated with concentrations of cell-bound nodularin. However, Aphanizomenon and Anabaena, two genera commonly co-occurring with Nodularia, also contain these pigments and thus the significant correlations obtained presumably originate from Nodularia being the dominant cyanobacterium in all samples collected in 2002.     

Proteomic profiling of the Baltic Sea cyanobacterium Nodularia spumigena strain AV1 during ammonium supplementation

The cyanobacterium Nodularia spumigena dominates the annual, toxic summer blooms in the Baltic Sea. Although Nodularia has been receiving attention due to its production of the hepatotoxin nodularin, molecular data regarding the regulation of nitrogen fixation is lacking. We have previously reported that N. spumigena strain AV1, unlike model filamentous cyanobacteria, differentiates heterocysts in the absence of detectable nitrogen fixation activity. To further analyze the uncoupling between these two linked processes, we assessed the impact of ammonium ions on the N. spumigena metabolism using a proteomic approach. Proteomic profiling was performed at three different times during ammonium supplementation using quantitative 2-dimensional gel electrophoresis followed by MS/MS analysis. Using this approach, we identified 34 proteins, 28 of which were unique proteins that changed successively in abundance during growth on ammonium. Our results indicate that N. spumigena generally exhibits lower energy production and carbon fixation in the presence of ammonium and seems to be inefficient in utilizing ammonium as an external nitrogen source. The possibility of ammonium toxicity due to PSII damage was investigated and the results are discussed. Our findings have implications in regard to the strategies considered to manage the cyanobacterial blooms in the Baltic Sea.     

Occurrence of the hepatotoxic cyanobacterium Nodularia spumigena in the Baltic Sea and structure of the toxin.

Water blooms formed by potentially toxic species of cyanobacteria are a common phenomenon in the Baltic Sea in late summer. Twenty-five cyanobacterial bloom samples were collected from open and coastal waters of the Baltic Sea during 1985 to 1987, and their toxicity was determined by mouse bioassay. All of 5 bloom samples from the southern Baltic Sea, 6 of 6 from the open northern Baltic Sea (Gulf of Finland), and 7 of 14 Finnish coastal samples were found to contain hepatotoxic cyanobacteria. Nodularia spumigena and Aphanizomenon flos-aquae occurred together in high amounts in blooms from the open-sea areas. In addition, coastal samples contained the species Anabaena lemmermannii, Microcystis aeruginosa, and Oscillatoria agardhii. Eighteen hepatotoxic N. spumigena cultures were isolated from water bloom and open-sea water samples. High-pressure liquid chromatographic analysis of both hepatotoxic bloom samples and Nodularia strains showed a single toxic fraction. The toxin concentrations of the blooms were less than or equal to 2.4 mg/g of freeze-dried material, and those of laboratory-grown cultures were 2.5 to 8.0 mg/g of freeze-dried cells. A single toxin was isolated from three N. spumigena-containing bloom samples and three N. spumigena laboratory isolates. Amino acid analysis and low- and high-resolution fast-atom bombardment mass spectroscopy indicated that the toxin from all of the sources was a cyclic pentapeptide (molecular weight, 824) containing glutamic acid, beta-methylaspartic acid, arginine, N-methyldehydrobutyrine, and 3-amino-9-methoxy-2,6,8-trimethyl-10-phenyl-4,6-decadienoic acid.(ABSTRACT TRUNCATED AT 250 WORDS)     

Molecular identification of bacteria associated with filaments of Nodularia spumigena and their effect on the cyanobacterial growth

Colonial and filamentous cyanobacteria frequently have bacteria associated with their extracellular mucus zone or more tightly attached to their cells surface. The toxin-producing cyanobacterium Nodularia spumigena is an important component of the Baltic Sea plankton community, and its filaments are likely to provide a microenvironment suitable for the development of a particular bacteria flora. In the present work, 13 bacterial strains associated with filaments of N. spumigena from the Baltic Sea were isolated and identified by sequencing the 16S rRNA gene. Different bacterial lineages were found associated with the cyanobacterial filaments, including the alpha, beta, and gamma subdivisions of the class Proteobacter and the division Firmicutes (Gram-positive bacteria). Several 16S rRNA gene sequences were not closely related to previously reported sequences of cultured bacteria from the Baltic Sea or to any other reported sequence. Conversely, sequences related to the gamma Proteobacter genus Shewanella, a group previously described in the Baltic Sea, were found among the isolates. The bacterial isolates were grown and added to cultures of exponentially growing N. spumigena. Five isolates, related to the alpha and gamma Proteobacter and Firmicutes, affected negatively the cyanobacterial growth, leading to a lower biomass yield up to 38% relative to controls with no bacteria addition. Five gamma Proteobacter-related strains had no effect on the cyanobacterial growth, while three strains related to Shewanella baltica had a positive effect. Although none of the bacterial isolates showed strong algicidal effect, the observed stimulatory and retarding effects on N. spumigena growth under culture conditions denotes the importance of the associated bacterial community for the dynamics of these cyanobacterial populations in nature. Moreover, several new taxa recovered in this study probably belong to species not yet described.     

Phytoplankton vertical distributions and composition in Baltic Sea cyanobacterial blooms

We studied the vertical structure of the phytoplankton community in two toxic cyanobacterial blooms in the offshore Baltic Sea. In 1994, vertically separated potentially toxic, diazotrophic and mixotrophic species (belonging to Cyanophyceae, Dinophyceae and Prymnesiophyceae) dominated. In 1997, picocyanobacteria, mainly in colonies, made up 40–50% of the total phytoplankton carbon biomass in the top 20 m both day and night. Colony-forming species of picocyanobacteria seem to be occasionally important and hitherto underestimated in the Baltic Sea.We found species-specific depth distribution patterns. Nodularia spumigena and Anabaena spp. were observed mainly above 10 m depth, while Aphanizomenon sp. was mostly found deeper, especially at night. Dinophysis norvegica was only abundant near the seasonal pycnocline and showed very limited diurnal migration. Other flagellates, including small Cryptophyceae and 10 identified Chrysochromulina species, occurred down to 40 m depth. Their vertical migration may help to retrieve nutrients from below the summer pycnocline.We conclude that considerable differences in dominating functional groups may occur between years/bloom stages, and that the vertical distribution pattern of many species is recurring at similar environmental conditions, suggesting species-specific niche-separation.     

Seasonal variability in the individual carbon content of the calanoid copepod Acartia bifilosa from the northern Baltic Sea

The seasonal variation in the carbon content of the calanoid copepod Acartia bifilosa was studied in the northern Baltic Sea. Monthly length-weight relationships were followed from November 1990 to October 1991 by analysing the organic carbon content of individuals, using a high temperature combustion method. The monthly length (L) on carbon (C) regressions of copepodites were best described by power functions (C = aL^b), where 75% to 88% of the variation could be explained by length. Nauplii length explained less of their carbon content (63–71 %). The carbon content per length was highest in summer when the nutritional situation was good and temperature at its highest. However, the carbon-length regressions differed significantly between months, except in winter, when two subsequent months had similar regressions. The results pointed out the importance of seasonal variation as well as the risk of making errors if biomass estimates based on length on carbon regressions are used. Carbon analysis should, if possible, be done on every sample or should at least be tested as to whether the regressions are consistent with the study material before estimations are calculated.     

Steryl chlorin esters in sediments of the southern Baltic Sea

Sediments of the southern Baltic Sea were analysed for content of steryl chlorin esters, the chlorin compounds discovered recently in the marine environment. The chlorin esters occur in the Baltic sediments in substantial amounts and form a considerable fraction of the total chlorin content. Among the physicochemical parameters studied the highest correlation with the steryl chlorins showed organic carbon content in sediments and the content of fraction of sediments smaller than 10 μm. A significant correlation was observed between the steryl chlorins content and other chlorins as chlorophylla, phaeophytina, pyrophaeophytina as well as with β-carotene, the distinctly less significant correlation was with phaeophorbidea. This indicates an other way of formation of the steryl chlorins from algae than zooplankton grazing.     

Influence of the Thin Drift Algal Mats on the Distribution of Macrozoobenthos in Kõiguste Bay, NE Baltic Sea

Macrozoobenthic communities within and outside of the drift algal mats were compared in Kõiguste Bay, NE Baltic Sea. The patches of the drift algae were on average 0.5–1 km wide in diameter covering about 25% of the total bottom area of the bay. Thickness of the mat did not exceed 6 cm. The biomass of the mat varied between 35 and 1391 g dw m^?2. The drift algal mats had no clear negative effect on macrozoobenthos except for a few infaunal species. The drift algae favoured several detrivorous, herbivorous and carnivorous species. Among the studied variables, the thickness of algal mat and oxygen concentration at near-bottom layer explained the best the structure of macrozoobenthos. Total number of invertebrate species increased curvilinearly with the thickness of algal mat having the peak value at 3–5 cm thick algal mat. To conclude, moderate drift algal mats increased habitat complexity and, thus, the diversity of benthic faunal assemblages in otherwise poorly vegetated coastal areas.     

Localization of Fe-containing superoxide dismutase in cyanobacteria from the Baltic Sea: depth and light dependency

The abundance and cellular location of Fe-containing superoxide dismutase (Fe-SOD) in trichomes of Nodularia , Aphanizomenon and Anabaena collected from various depths in the Baltic Sea, and in trichomes of a cultured Nodularia strain, BC Nod-9427, isolated from the Baltic Sea, was examined by immunogold labelling. For trichomes collected from natural populations the areal concentration of Fe-SOD labelling decreased with depth: trichomes collected from surface accumulations had between 8 and 11 gold particles μm⁻² whereas trichomes collected from a depth of 18 m were unlabelled. When trichomes collected from a depth of 10 m (mean areal labelling density 0·5 gold particles μm⁻²) were exposed to the higher irradiances present at 1 m, the areal concentration of Fe-SOD increased to 3·5–4 gold particles μm⁻² within 4 h. When cultures of Nodularia strain BC Nod-9427, adapted to low light (10 μmol m⁻² s⁻¹), were transferred to an incident irradiance of 1350 μmol m⁻² s⁻¹, a doubling of the areal concentration of Fe-SOD gold label was observed within 1 h. Addition of 3-(3,4-dichlorophenyl)-1,1'-dimethylurea (DCMU) to cultures immediately before their transfer to increased illumination resulted in a decrease in areal Fe-SOD concentrations whereas addition of CdCl₂ caused an increase over and above that induced by the elevated irradiance. These results suggest that Baltic Sea cyanobacteria are able to modulate their Fe-SOD content but that this might be in response to oxidative stress rather than to light per se .