Tumor promoting effects of cyanobacterial extracts are potentiated by anthropogenic contaminants--evidence from in vitro study

Inhibition of gap junctional intercellular communication (GJIC) is affiliated with tumor promotion process and it has been employed as an in vitro biomarker for evaluation of tumor promoting effects of chemicals. In the present study we investigated combined effects of anthropogenic environmental contaminants 2,2′,4,4′,5,5′-hexachlorobiphenyl (PCB 153) and fluoranthene, cyanotoxins microcystin-LR and cylindrospermopsin, and extracts of laboratory cultures of cyanobacteria Aphanizomenon gracile and Cylindrospermopsis raciborskii, on GJIC in the rat liver epithelial cell line WB-F344. Binary mixtures of PCB 153 with fluoranthene and the mixtures of the two cyanobacterial strains elicited simple additive effects on GJIC after 30 min exposure, whereas microcystin-LR and cylindrospermopsin neither inhibited GJIC nor altered effects of PCB 153 or fluoranthene. However, synergistic effects were observed in the cells exposed to binary mixtures of anthropogenic contaminants (PCB 153 or fluoranthene) and cyanobacterial extracts. The synergistic effects were especially pronounced after prolonged (6-24 h) co-exposure to fluoranthene and A. gracile extract, when mixture caused nearly complete GJIC inhibition, while none of the individual components caused any downregulation of GJIC at the same concentration and exposure time. The effects of cyanobacterial extracts were independent of microcystin-LR or cylindrospermopsin, which were not detected in cyanobacterial biomass. It provides further evidence on the presence of unknown tumor promoting metabolites in cyanobacteria. Clear potentiation of the GJIC inhibition observed in the mixtures of two anthropogenic contaminants and cyanobacteria highlight the importance of combined toxic effects of chemicals in complex environmental mixtures.     

Toxic risk associated with sporadic occurrences of Microcystis aeruginosa blooms from tidal rivers in marine and estuarine ecosystems and its impact on Artemia franciscana nauplii populations

Microcystis aeruginosa is a species of freshwater cyanobacteria which can form harmful algal blooms in freshwater water bodies worldwide. However, in spite its sporadic occurrences for short periods of time in estuarine waters, their influence on zooplankton populations present in these ecosystems has not been extensively studied. In this work, Artemia franciscana was used as test organism model, studying mortality against several strains of M. aeruginosa with different degrees of toxigenicity, measuring whole-live cells and homogenate extracts. Results were compared with microcystin-LR equivalent content, measured by immunoassay. The results show that there were no significant differences between both exposure models (whole cells and extracts), and there are significant differences respect to the toxigenicity of cyanobacterial blooms depending of the M. aerugionosa strain involved in the process. Analysis of microcystin-LR equivalent concentration test immediately below the lowest significant concentration in all M. aerugionosa strains was used to determine the potential risk associated with the cell densities during a bloom. Comparison among the selected M. aerugionsa strains show that these factors have influence in the results obtained, and thus, several differences have been evidenced depending of the microcystin-LR equivalent production and the strain type involved.     

Impact of the toxicity of Cylindrospermopsis raciborskii (Woloszynska) Seenayya & Subba Raju on laboratory rats in vivo

In vivo laboratory studies of toxicity were performed on Wistar rats using a methanol extract produced by the natural population of Cylindrospermopsis raciborskii (abundance of 2.13 × 10⁵ trichomes mL⁻¹) collected at Aleksandrovac Lake (Serbia). HPLC analysis showed that the extract contains 6.65 μg cylindrospermopsin (CYN) mg⁻¹. The rats were killed 24 or 72 h after a single intraperitoneal injection of C. raciborskii extract in concentrations of 1500, 3000, 6000 and 12,000 μg kg⁻¹ body weight (bw) and an equivalent amount of CYN as present in the highest dose of the extract (79.80 μg CYN kg⁻¹ bw). The genotoxic effect on the livers treated with C. raciborskii was evaluated using comet assay and potential induction of oxidative stress as the toxicity mechanism associated with the presence of CYN in extract. The results from the analyses of DNA damage in the comet tail length, tail moment and percentage of DNA in the tail in the liver indicated that administration of extract and CYN present statistically significant difference when compared with the negative control group. Although an increase in the frequency of selected parameters induced by the CYN was observed in the liver, this damage was less than the damage resulting from the administration of the highest dose of extract. The changes in the biochemical parameters of the hepatic damage showed that the application of single doses of the extract and CYN did not cause serious liver damage in rats. The extract and CYN significantly increased oxidative stress in rats’ liver after a single exposure.

     

A rapid bioassay for detecting saxitoxins using a Daphnia acute toxicity test

Bioassays using Daphnia pulex and Moina micrura were designed to detect cyanobacterial neurotoxins in raw water samples. Phytoplankton and cyanotoxins from seston were analyzed during 15 months in a eutrophic reservoir. Effective time to immobilize 50% of the exposed individuals (ET₅₀) was adopted as the endpoint. Paralysis of swimming movements was observed between ∼0.5-3 h of exposure to lake water containing toxic cyanobacteria, followed by an almost complete recovery of the swimming activity within 24 h after being placed in control water. The same effects were observed in bioassays with a saxitoxin-producer strain of Cylindrospermopsis raciborskii isolated from the reservoir. Regression analysis showed significant relationships between ET₅₀vs. cell density, biomass and saxitoxins content, suggesting that the paralysis of Daphnia in lake water samples was caused by saxitoxins found in C. raciborskii. Daphnia bioassay was found to be a sensitive method for detecting fast-acting neurotoxins in natural samples, with important advantages over mouse bioassays.     

Survey of cyanobacterial toxins in Czech water reservoirs—the first observation of neurotoxic saxitoxins

The environmental occurrence and concentrations of cyanobacterial toxins (cyanotoxins) were investigated in the Czech Republic. Concentrations of microcystins (MCs), cylindrospermopsin (CYN) or saxitoxins (STXs) were determined immunochemically by ELISA assays in 30 water samples collected from the surface layers of 19 reservoirs during the summer season of 2010. MCs were detected in 18 reservoirs and 83 % of samples, with median and maximal concentration being 1.5 and 18.6 μg/L, respectively. The high frequency of MC occurrence coincided with prevalence of cyanobacterium Microcystis sp., which was detected in 87 % samples, followed by Dolichospermum (Anabaena) sp. observed in 33 % samples. CYN was detected by ELISA only in one sample at a concentration of 1.2 μg/L. STXs presence was indicated for the first time in Czech water reservoirs when the toxins were found at low concentrations (0.03–0.04 μg/L) in two samples (7 %) collected from two different reservoirs, where STXs co-occurred with MCs and eventually also with CYN. In both STX-positive samples, the phytoplankton community was dominated by Microcystis sp., but Dolichospermum sp. and/or Aphanizomenon sp. were also present as putative producers of STX and/or CYN. Cyanotoxins commonly occurred in Czech water reservoirs, and MCs frequently at concentrations possibly associated with human health risks. MCs were the most prevalent and abundant cyanotoxins, but also other cyanotoxins were detected, though sporadically. Further research and regulatory monitoring of cyanotoxins other than MCs is therefore required.     

Remote estimation of phycocyanin (PC) for inland waters coupled with YSI PC fluorescence probe

Nuisance cyanobacterial blooms degrade water resources through accelerated eutrophication, odor generation, and production of toxins that cause adverse effects on human health. Quick and effective methods for detecting cyanobacterial abundance in drinking water supplies are urgently needed to compliment conventional laboratory methods, which are costly and time consuming. Hyperspectral remote sensing can be an effective approach for rapid assessment of cyanobacterial blooms. Samples (n?=?250) were collected from five drinking water sources in central Indiana (CIN), USA, and South Australia (SA), which experience nuisance cyanobacterial blooms. In situ hyperspectral data were used to develop models by relating spectral signal with handheld fluorescence probe (YSI 6600 XLM-SV) measured phycocyanin (PC in cell/ml), a proxy pigment unique for indicating the presence of cyanobacteria. Three-band model (TBM), which is effective for chlorophyll-a estimates, was tuned to quantify cyanobacteria coupled with the PC probe measured cyanobacteria. As a comparison, two band model proposed by Simis et al. (Limnol Oceanogr, 50(11): 237–245, 2005; denoted as SM05) was paralleled to evaluate TBM model performance. Our observation revealed a high correlation between measured and estimated PC for SA dataset (R ²?=?0.96; range: 534–20,200 cell/ml) and CIN dataset (R ²?=?0.88; range: 1,300–44,500 cell/ml). The potential of this modeling approach for imagery data were assessed by simulated ESA/Centinel3/OLCI spectra, which also resulted in satisfactory performance with the TBM for both SA dataset (RMSE %?=?26.12) and CIN dataset (RMSE %?=?34.49). Close relationship between probe-measured PC and laboratory measured cyanobacteria biovolume was observed (R ²?=?0.93, p?<?0.0001) for the CIN dataset, indicating a stable performance for PC probe. Based on our observation, field spectroscopic measurement coupled with PC probe measurements can provide quantitative cyanobacterial bloom information from both relatively static and flowing inland waters. Hence, it has promising implications for water resource managers to obtain information for early warning detection of cyanobacterial blooms through the close association between probe measured PC values and cyanobacterial biovolume via remote sensing modeling.     

Biodegradation of multiple cyanobacterial metabolites in drinking water supplies

The fate of multiple cyanobacterial metabolites was assessed in two Australian source waters. The saxitoxins were the only metabolites shown to be non-biodegradable in Myponga Reservoir water, while microcystin-LR (MCLR) and geosmin were biodegradable in this water source. Likewise, cylindrospermopsin (CYN) was shown to be biodegradable in River Murray water. The order of ease of biodegradability followed the trend: MCLR > CYN > geosmin > saxitoxins. Biodegradation of the metabolites was affected by temperature and seasonal variations with more rapid degradation at 24 °C and during autumn compared with 14 °C and during winter. A microcystin-degrading bacterium was isolated and shown to degrade four microcystin variants within 4 h. This bacterium, designated as TT25, was shown to be 99% similar to a Sphingopyxis sp. based on a 16S rRNA gene fragment. Isolate TT25 was shown to contain a homologue of the mlrA gene; the sequence of which was 99% similar to that of a previously reported microcystin-degrader. Furthermore, isolate TT25 could degrade the microcystins in the presence of copper sulphate (0.5 mg L⁻¹ as Cu²⁺) which is advantageous for water authorities dosing such algicides into water bodies to control cyanobacterial blooms.     

Identification of microcystins from three collection strains of Microcystis aeruginosa

Microcystins (MCs) are toxic cyclic heptapeptides produced by various cyanobacteria genera, especially Microcystis. We identified 10 out of 12 MCs produced by three Microcystis aeruginosa strains from cyanobacteria collections, UTEX 2666, UTEX 2670 and UAM 1303, by using two analytical methods: Matrix-assisted Laser Desorption Ionization Time of Flight Mass Spectrometry (MALDI-TOF/MS) and HPLC Photodiode Array Detector coupled to a hybrid Quadrupole Time of Flight Mass Spectrometry (HPLC-PDA-QTOF/MS). MALDI-TOF/MS failed to detect non-polar MCs, such as MC-LY and MC-LW. HPLC-QTOF/MS permitted the accurate identification of most MCs present in methanolic extracts. Besides, three new MCs, namely: [D-Glu(OCH₃)⁶, D-Asp³] MC-LAba, MC-YL and MC-YM were detected by HPLC-QTOF/MS.     

Eutrophic urban ponds suffer from cyanobacterial blooms: Dutch examples

Ponds play an important role in urban areas. However, cyanobacterial blooms counteract the societal need for a good water quality and pose serious health risks for citizens and pets. To provide insight into the extent and possible causes of cyanobacterial problems in urban ponds, we conducted a survey on cyanobacterial blooms and studied three ponds in detail. Among 3,500 urban ponds in the urbanized Dutch province of North Brabant, 125 showed cyanobacterial blooms in the period 2009–2012. This covered 79 % of all locations registered for cyanobacterial blooms, despite the fact that urban ponds comprise only 11 % of the area of surface water in North Brabant. Dominant bloom-forming genera in urban ponds were Microcystis, Anabaena and Planktothrix. In the three ponds selected for further study, the microcystin concentration of the water peaked at 77 μg l^?1 and in scums at 64,000 μg l^?1, which is considered highly toxic. Microcystin-RR and microcystin-LR were the most prevalent variants in these waters and in scums. Cyanobacterial chlorophyll-a peaked in August with concentrations up to 962 μg l^?1 outside of scums. The ponds were highly eutrophic with mean total phosphorus concentrations between 0.16 and 0.44 mg l^?1, and the sediments were rich in potential releasable phosphorus. High fish stocks dominated by carp lead to bioturbation, which also favours blooms. As urban ponds in North Brabant, and likely in other regions, regularly suffer from cyanobacterial blooms and citizens may easily have contact with the water and may ingest cyanobacterial material during recreational activities, particularly swimming, control of health risk is of importance. Monitoring of cyanobacteria and cyanobacterial toxins in urban ponds is a first step to control health risks. Mitigation strategies should focus on external sources of eutrophication and consider the effect of sediment P release and bioturbation by fish.     

Alterations observed in the endothelial HUVEC cell line exposed to pure Cylindrospermopsin

The cyanobacterial toxin Cylindrospermopsin (CYN) is receiving great interest due to its increasing presence in waterbodies, which has lead to recognize it as a potential threat to drinking water safety. CYN is a potent inhibitor of protein and glutathione synthesis. The present work studies for the first time the effects of CYN in endothelial cells. The basal cytotoxicity endpoints studied at 24 and 48 h were total protein content (PC), neutral red (NR) uptake and the tretazolium salt, MTS, reduction. Moreover, the effect of subcytotoxic concentrations of CYN on the generation of intracellular reactive oxygen species (ROS), the activity of γ-glutamylcysteine synthetase (GCS) and glutathione (GSH) content have been investigated. In addition, morphological alterations of HUVEC cells subsequent to CYN exposure were recorded.The cytotoxicity endpoints revealed a decrease in the cellular viability in a time and concentration-dependent way. The most sensitive cytotoxicity endpoint was NR uptake assay, with reductions in cell viability of 95% at 48 h of exposure to 40 μg mL⁻¹ CYN. Intracellular ROS production was increased only at the lowest concentration assayed, while GCS activity and GSH content underwent concentration-dependent enhancements. The most remarkable morphological alterations observed were: nucleolar segregation with altered nuclei, degenerated Golgi apparatus, increases in the presence of granules and apoptosis.     

Identification and characterization of coagulation inhibitor proteins derived from cyanobacterium Microcystis aeruginosa

The excess growth of cyanobacteria in semi enclosed water areas caused by eutrophication brings about coagulation inhibition in drinking water treatment processes. In this study, coagulation inhibitor proteins produced by Microcystis aeruginosa, a major cyanobacterium in algal bloom, were acquired by a phage display technique, an aluminum-immobilized affinity chromatography and a protein expression technique using Escherichia coli cells. Two cyanobacterial peptides with a high ratio of metallophilic amino acids were recovered, which were a part of homologues of a thiol oxidase enzyme Ero1p and a trans-acting repressor ArsR. It was also shown that the homologue of ArsR exhibited a stronger inhibitory effect on the coagulation of kaolin suspension with polyaluminum chloride than the control proteins. This is the first report to identify a cyanobacterial cell component to inhibit coagulation. The compositions of polar amino acids were critical to explain the strength of coagulation inhibition potential. Polar proteins from cyanobacteria could collectively consume coagulants or stabilize clay particles, which would be plausible explanations for causing coagulation inhibition. Meanwhile, results from the kaolin coagulation tests using the control proteins implied that the neutralization of positive charges of coagulant constituents by simple electrostatic interactions might not be the key mechanism on the protein-induced coagulation inhibition.     

Differential Sensitivity of a Coccal Green Algal and a Cyanobacterial Species to Dissolved Natural Organic Matter (NOM) (8 pp)

Background

and Aim. In non-eutrophicated freshwaters, humic substances (HS) pose chemical stresses on aquatic organisms and, hence, separating sensitive from less sensitive or even tolerant species. One of the stresses, identified so far, is the reduction of photosynthetic oxygen production and reduction in growth in freshwater macrophytes and algae. In a previous paper, it has been shown that even closely related coccal green algae responded differently upon identical stress by HS, which is consistent with the hypothesis above. Due to their much simpler cellular ultrastructure, cyanobacteria are supposed to be more sensitive to HS exposure than eukaryote should be. One coccal green algal species (Desmodesmus communis) and one cyanobacterium (Chroococcus minutus) were exposed to four natural organic matter (NOM) isolates. One NOM has been isolated from a brown water lake (Schwarzer See) in Brandenburg State; three were obtained from a comprehensive Scandinavian NOM research project and originated from Norway (Birkenes), Finland (Hietajärvi), and Sweden (Svartberget).

Methods

Cultures of D. communis and C. minutus were obtained from the Culture Collection of Algae, Göttingen, and maintained in a common medium. The cultures were non-axenic. The algae and cyanobacteria were exposed under identical conditions to environmentally realistic NOM concentrations. Cell numbers were counted microscopically in Neugebauer cuvettes in triplicates. To avoid limitation by nutrient depletion, the experiments were terminated after 14 to 15 days. Until culture day 12, no growth limitation has been observed in the controls.

Results

All NOM isolates modulated the growth of the algae and cyanobacteria. During the early culture days, there was a slight growth promotion with the coccal green alga and to a much lesser degree with the cyanobacterium. Yet, the major effect were significant reductions in cell yield in both primary producer cultures. C. minutus was much more affected than D. communis. This applies particularly to the three tested Scandinavian NOM isolates, which were effective at concentrations even below 1 mg L-1 DOC.

Discussion

The growth promoting effect may be due to an increase in bioavailability of some trace nutrients in the presence of NOM, the release of some growth promoting substances by microbial or photochemical processing of the NOM, and/or a hormetic effect. The growth reducing effect can be explained as a herbicide-like mode of action that affects the photosystem II most prevalent by blocking the electron transport chain, absorption of electrons, or production of an internal oxidative stress after processing the bioconcentrated HS. Furthermore, it may be postulated that also photo-toxicity of these HS in the algal cells contributes to the overall toxicity; however, experimental evidence is lacking so far.

Conclusions

Upon exposure to HS, cyanobacteria appear to be much more sensitive than coccal green algae and respond in growth reduction. This high sensitivity of cyanobacteria to HS may explain phytoplankton patterns in the field. Eutrophic, humic-rich lakes do not support the cyanobacterial blooms characteristic of eutrophic, but humic-poor lakes. In the humic-rich systems, raphidophytes or, less frequent, specific coccal greens are more common. Obviously, cyanobacteria appear to be unable to make advantage of their accessory pigments (phycocyanin) to exploit the reddish light prevailing in humic-rich lakes.

Recommendation

. At present, no effective structure can be figured out which may be responsible for the adverse effect on the cyanobacterial species. It is reserved to future research whether or not HS may be applied more specifically (for instance, with elevated moieties of the effective structures) as a natural geochemical to combat cyanobacterial blooms.

     

Effects of suspended solids on cyanobacterial bloom formation under different wind fields

Wind waves and suspended solids (SS) generated by the resuspension of sediments are ubiquitous characteristics of lake ecosystems. However, their effects on phytoplankton remain poorly elucidated in shallow eutrophic lakes. Laboratory experiments were carried out to investigate the responses of Microcystis aeruginosa to SS under static (wind speed of 0 m/s) and breeze (wind speed of 3 m/s) conditions. Results showed that 50 mg/L SS can promote the growth of M. aeruginosa, accelerate the formation of colonies, and increase the floating rate under no-wind conditions. Comparing with static environment, breeze can significantly increase the growth rate of M. aeruginosa and benefit the formation of larger colonies of algae cells. Driven by wind and SS, the buoyancy of the cyanobacteria community in different experimental groups was obviously different. The specific performance was that low SS concentration and breeze were in favor of the floating of cyanobacteria, while high SS concentration went against the floating of algal cells. As a conclusion, wind speed of 3 m/s and 20–50 mg/L SS have a synergistic effect on the formation of cyanobacterial blooms. This study can provide an improved current understanding of bloom formation and turbidity management strategies in shallow eutrophic lakes.

     

On the successful use of a simplified model to simulate the succession of toxic cyanobacteria in a hypereutrophic reservoir with a highly fluctuating water level

Many freshwater bodies worldwide that suffer from harmful algal blooms would benefit for their management from a simple ecological model that requires few field data, e.g. for early warning systems. Beyond a certain degree, adding processes to ecological models can reduce model predictive capabilities. In this work, we assess whether a simple ecological model without nutrients is able to describe the succession of cyanobacterial blooms of different species in a hypereutrophic reservoir and help understand the factors that determine these blooms. In our study site, Karaoun Reservoir, Lebanon, cyanobacteria Aphanizomenon ovalisporum and Microcystis aeruginosa alternatively bloom. A simple configuration of the model DYRESM-CAEDYM was used; both cyanobacteria were simulated, with constant vertical migration velocity for A. ovalisporum, with vertical migration velocity dependent on light for M. aeruginosa and with growth limited by light and temperature and not by nutrients for both species. The model was calibrated on two successive years with contrasted bloom patterns and high variations in water level. It was able to reproduce the measurements; it showed a good performance for the water level (root-mean-square error (RMSE) lower than 1 m, annual variation of 25 m), water temperature profiles (RMSE of 0.22–1.41 °C, range 13–28 °C) and cyanobacteria biomass (RMSE of 1–57 μg Chl a L^?1, range 0–206 μg Chl a L^?1). The model also helped understand the succession of blooms in both years. The model results suggest that the higher growth rate of M. aeruginosa during favourable temperature and light conditions allowed it to outgrow A. ovalisporum. Our results show that simple model configurations can be sufficient not only for theoretical works when few major processes can be identified but also for operational applications. This approach could be transposed on other hypereutrophic lakes and reservoirs to describe the competition between dominant phytoplankton species, contribute to early warning systems or be used for management scenarios.     

Effects of Bacillus subtilis on the growth,colony maintenance,and attached bacterial community composition of colonial cyanobacteria

In freshwater aquaculture ponds, application of algicidal Bacillus is a promising way in the control of cyanobacterial blooms. To best understand Bacillus algicidal characters and mechanisms in the field, different-sized colonial cyanobacteria were isolated from an aquaculture pond, and the effects of B. subtilis on their growth, colony maintenance, and colony-attached bacterial community composition were investigated. The results showed that B. subtilis could inhibit the growth of colonial cyanobacteria. Bigger-sized colonies isolated from the field could spontaneously disintegrate into smaller-sized colonies in the laboratory. Algicidal B. subtilis could accelerate the disintegration of colonies and decrease colony size. B. subtilis not only decreased the colony-attached bacterial community diversity but also changed its composition. B. subtilis increased the relative abundances of some attached bacterial genera, including Pseudomonas, Shewanella, Bacillus, Shinella, Rhizobium, and Ensifer. These bacteria with algicidal, microcystin-degrading, and flocculating activities might be an important contributor to algicidal effects of B. subtilis on colonial cyanobacteria.

     

Influence of alum on cyanobacterial blooms and water quality of earthen fish ponds

Eruption of blue-green algal blooms occurs frequently in eutrophic lakes and fish ponds, with associated unpleasant odor and horrid scums. In the present study, we conducted a pre-test experiment in 3 m³ outdoor concrete ponds to determine the optimum concentration of aluminum sulfate (alum) required for reduction of the cyanobacterial blooms without negative effect on fish growth. As a consequence, 10 mg L^?1 alum was named as the optimum concentration that was applied in 1000 m³ earthen fish ponds. Obtained results showed that Secchi disc values significantly increased from 10 to 24 cm after 14 days of alum application. Alum-treated ponds showed a reduction in total phytoplankton counts by 94 and 96 % compared to the corresponding controls after 10 and 14 days, respectively. Abundance of blue-green algae in the treated ponds was decreased by 98 % compared to the corresponding control after 14 days of alum application. Consequently, dissolved oxygen, pH, total phosphorus, orthophosphate, and chlorophyll “a” content declined significantly. Our study revealed that using 10 mg L^?1 of alum is an effective way to control cyanobacterial blooms in eutrophic waters, especially in fish ponds, without negative effect in water quality.     

Nitrogen fixation by cyanobacteria stimulates production in Baltic food webs

Filamentous, nitrogen-fixing cyanobacteria form extensive summer blooms in the Baltic Sea. Their ability to fix dissolved N₂ allows cyanobacteria to circumvent the general summer nitrogen limitation, while also generating a supply of novel bioavailable nitrogen for the food web. However, the fate of the nitrogen fixed by cyanobacteria remains unresolved, as does its importance for secondary production in the Baltic Sea. Here, we synthesize recent experimental and field studies providing strong empirical evidence that cyanobacterial nitrogen is efficiently assimilated and transferred in Baltic food webs via two major pathways: directly by grazing on fresh or decaying cyanobacteria and indirectly through the uptake by other phytoplankton and microbes of bioavailable nitrogen exuded from cyanobacterial cells. This information is an essential step toward guiding nutrient management to minimize noxious blooms without overly reducing secondary production, and ultimately most probably fish production in the Baltic Sea.     

Temperature and nutrients are significant drivers of seasonal shift in phytoplankton community from a drinking water reservoir,subtropical China

Reservoirs are an important source of water supply in many densely populated areas in southeast China. Phytoplankton plays an important role in maintaining the structure and function of these reservoir ecosystems. Understanding of seasonal succession in phytoplankton communities and its driving factors is essential for effective water quality management in drinking-water reservoirs. In this study, water samples were collected monthly at the surface layers of riverine, transitional, and lacustrine zones from May 2010 to April 2011 in Tingxi Reservoir, southeast China. The phytoplankton showed distinct seasonal shifts in community structure at both taxonomic and functional levels. Cyanophyta was the dominant group in summer, especially species of Raphidiopsis in May and Aphanizomenon in June, and cyanobacterial dominance was promoted by both warmer conditions and excessive nutrients loading. Cyanophyta was gradually replaced by Cryptophyta (e.g., Chroomonas caudata) in abundance and by Bacillariophyta (Fragilaria sp. or Synedra sp. and Melosira sp.) in biomass with decreasing temperature. It appeared that seasonal shifts in phytoplankton composition were closely related to climate, nutrient status, and hydrology in this reservoir. Our partial RDA results clearly showed that water temperature and nutrients (TN and TP) were the most critical factors driving phytoplankton community shift in the abundance and biomass data, respectively. Further, with the global warming, cyanobacterial blooms may increase in distribution, duration, and intensity. In our study, the abundance and biomass of cyanobacteria had significant and positive correlations with temperature and phosphorus. Therefore, a stricter limit on nutrient input should be a priority in watershed management to protect drinking water from the effects of cyanobacterial blooms, especially in high-temperature period.     

An integrated method for removal of harmful cyanobacterial blooms in eutrophic lakes

As the eutrophication of lakes becomes an increasingly widespread phenomenon, cyanobacterial blooms are occurring in many countries. Although some research has been reported, there is currently no good method for bloom removal. We propose here a new two-step integrated approach to resolve this problem. The first step is the inactivation of the cyanobacteria via the addition of H(2)O(2). We found 60 mg/L was the lowest effective dose for a cyanobacterial concentration corresponding to 100 μg/L chlorophyll-a. The second step is the flocculation and sedimentation of the inactivated cyanobacteria. We found the addition of lake sediment clay (2 g/L) plus polymeric ferric sulfate (20 mg/L) effectively deposited them on the lake bottom. Since algaecides and flocculants had been used separately in previous reports, we innovatively combined these two types of reagents to remove blooms from the lake surface and to improve the dissolved oxygen content of lake sediments.     

Shewanella spp. from wastewater treatment plant-affected environment: isolation and characterization

Bacteria from the genus Shewanella are inhabitants of marine and freshwater ecosystems, recognized fish spoilage bacteria, but less known as fish disease agents. Shewanella spp. isolated from fish living in waters close to effluents of a wastewater treatment plant (WWTP) were not previously characterized. We have tested Shewanella isolates from WWTP-affected waters and related fish. Genotypic characterization identified most strains as S. baltica and S. oneidensis. In order to investigate the sensibility and accuracy of their MALDI-TOF MS identification, they were grown on two culture media enriched by various NaCl concentrations, incubated at different temperatures and duration. We analyzed their antimicrobial susceptibility on a panel of antimicrobial drugs and capacity for biofilm production. With a view to demonstrate their capacity to produce fatty acids, we assessed the impact of different culture media on their lipid profile. We performed zebrafish embryotoxicity tests to simulate the environmental infection of the earliest life stages in S. baltica-contaminated waters. The best MALDI-TOF MS identification scores were for strains cultivated on TSA for 24 h at 22 °C and with supplementation of 1.5% NaCl. Less than 17% of isolates demonstrated antimicrobial resistance. Most isolates were weak biofilm producers. Strain-to-strain variation of MIC and MBC was low. The major fatty acids were C15:0, C16:0, C16:1, C17:1, and iC15:0. Exposure of Danio rerio to different S. baltica concentrations induced severe effects on zebrafish development: decreased heartbeat rate, locomotor activity, and melanin pigmentation. S. baltica passed through chorionic pores of zebrafish.