Evaluating microcystinase A-based approach on microcystins degradation during harvested cyanobacterial blooms.

Addressing notorious and worldwide Microcystis blooms, mechanical algae harvesting is an effective emergency technology for bloom mitigation and removal of nutrient loads in waterbodies. However, the absence of effective methods for removal of cyanobacterial toxins, e.g., microcystins (MCs), poses a challenge to recycle the harvested Microcystis biomass. In this study, we therefore introduced a novel approach, the "captured biomass-MlrA enzymatic MC degradation", by enriching microcystinase A (MlrA) via fermentation and spraying it onto salvaged Microcystis slurry to degrade all MCs. After storing the harvested Microcystis slurry, a rapid release of extracellular MCs occurred within the initial 8 h, reaching a peak concentration of 5.33 µg/mL at 48 h during the composting process. Upon spraying the recombinant MlrA crude extract (about 3.36 U) onto the Microcystis slurry in a ratio of 0.1% (v/v), over 95% of total MCs were degraded within a 24-h period. Importantly, we evaluated the reliability and safety of using MlrA extracts to degrade MCs. Results showed that organic matter/nutrient contents, e.g. soluble proteins, polysaccharides, phycocyanin and carotenoids, were not significantly altered. Furthermore, the addition of MlrA extracts did not significantly change the bacterial community composition and diversity in the Microcystis slurry, indicating that the MlrA extracts did not increase the risk of pathogenic bacteria. Our study provides an effective and promising method for the pre-treatment of harvested Microcystis biomass, highlighting an ecologically sustainable framework for addressing Microcystis blooms.

Changes in CO2 concentration drive a succession of toxic and non-toxic strains of Microcystis blooms.

The dynamic changes between toxic and non-toxic strains of Microcystis blooms have always been a hot topic. Previous studies have found that low CO2 favors toxic strains, but how changing dissolved CO2 (CO2 [aq]) in water body influences the succession of toxic and non-toxic strains in Microcystis blooms remains uncertain. Here, we combined laboratory competition experiments, field observations, and a machine learning model to reveal the links between CO2 changes and the succession. Laboratory experiments showed that under low CO2 conditions (100-150 ppm), the toxic strains could make better use of CO2 (aq) and be dominant. The non-toxic strains demonstrated a growth advantage as CO2 concentration increased (400-1000 ppm). Field observations from June to November in Lake Taihu showed that the percentage of toxic strains increased as CO2 (aq) decreased. Machine learning highlighted links between the inorganic carbon concentration and the proportion of advantageous strains. Our findings provide new insights for cyanoHABs prediction and prevention.

Changes in nitrogen metabolism of phosphorus-starved bloom-forming cyanobacterium Microcystis aeruginosa: Implications for nutrient management.

The surplus of nitrogen plays a key role in the maintenance of cyanobacterial bloom when phosphorus has already been limited. However, the interplay between high nitrogen and low phosphorus conditions is not fully understood. Nitrogen metabolism is critical for the metabolism of cyanobacteria. Transcriptomic analysis in the present study suggested that nitrogen metabolism and ribosome biogenesis were the two most significantly changed pathways in long-term phosphorus-starved bloom-forming cyanobacteria Microcystis aeruginosa FACHB-905. Notably, the primary glutamine synthetase/glutamate synthase cycle, crucial for nitrogen metabolism, was significantly downregulated. Concurrently, nitrogen uptake showed a marked decrease due to reduced expression of nitrogen source transporters. The content of intracellular nitrogen reservoir phycocyanin also showed a drastic decrease upon phosphorus starvation. Our study demonstrated that long-term phosphorus-starved cells also suffered from nitrogen deficiency because of the reduction in nitrogen assimilation, which might be limited by the reduced ribosome biogenesis and the shortage of adenosine triphosphate. External nitrogen supply will not change the transcriptions of nitrogen metabolism-related genes significantly like that under phosphorus-rich conditions, but still help to maintain the survival of phosphorus-starved cells. The study deepens our understanding about the survival strategies of Microcystis cells under phosphorus starvation and the mutual dependence between nitrogen and phosphorus, which would provide valuable information for nutrient management in the eutrophicated water body.

The facilitating role of phycospheric heterotrophic bacteria in cyanobacterial phosphonate availability and Microcystis bloom maintenance.

BACKGROUND: Phosphonates are the main components in the global phosphorus redox cycle. Little is known about phosphonate metabolism in freshwater ecosystems, although rapid consumption of phosphonates has been observed frequently. Cyanobacteria are often the dominant primary producers in freshwaters; yet, only a few strains of cyanobacteria encode phosphonate-degrading (C-P lyase) gene clusters. The phycosphere is defined as the microenvironment in which extensive phytoplankton and heterotrophic bacteria interactions occur. It has been demonstrated that phytoplankton may recruit phycospheric bacteria based on their own needs. Therefore, the establishment of a phycospheric community rich in phosphonate-degrading-bacteria likely facilitates cyanobacterial proliferation, especially in waters with scarce phosphorus. We characterized the distribution of heterotrophic phosphonate-degrading bacteria in field Microcystis bloom samples and in laboratory cyanobacteria "phycospheres" by qPCR and metagenomic analyses. The role of phosphonate-degrading phycospheric bacteria in cyanobacterial proliferation was determined through coculturing of heterotrophic bacteria with an axenic Microcystis aeruginosa strain and by metatranscriptomic analysis using field Microcystis aggregate samples. RESULTS: Abundant bacteria that carry C-P lyase clusters were identified in plankton samples from freshwater Lakes Dianchi and Taihu during Microcystis bloom periods. Metagenomic analysis of 162 non-axenic laboratory strains of cyanobacteria (consortia cultures containing heterotrophic bacteria) showed that 20% (128/647) of high-quality bins from eighty of these consortia encode intact C-P lyase clusters, with an abundance ranging up to nearly 13%. Phycospheric bacterial phosphonate catabolism genes were expressed continually across bloom seasons, as demonstrated through metatranscriptomic analysis using sixteen field Microcystis aggregate samples. Coculturing experiments revealed that although Microcystis cultures did not catabolize methylphosphonate when axenic, they demonstrated sustained growth when cocultured with phosphonate-utilizing phycospheric bacteria in medium containing methylphosphonate as the sole source of phosphorus. CONCLUSIONS: The recruitment of heterotrophic phosphonate-degrading phycospheric bacteria by cyanobacteria is a hedge against phosphorus scarcity by facilitating phosphonate availability. Cyanobacterial consortia are likely primary contributors to aquatic phosphonate mineralization, thereby facilitating sustained cyanobacterial growth, and even bloom maintenance, in phosphate-deficient waters. Video Abstract.

Non-targeted metabolomic profiling of filamentous cyanobacteria Aphanizomenon flos-aquae exposed to a concentrated culture filtrate of Microcystis aeruginosa.

Microcystis and Aphanizomenon are two toxic cyanobacteria genera, which frequently cause blooms in freshwater lakes. In some cases, succession of these two genera was observed in natural water bodies. Among the diverse factors contributing to such succession of dominant cyanobacterial genera, an allelopathic effect was proposed to be involved after the growth inhibitory effect of several Microcystis species on A. flos-aquae was investigated. However, the response of target species exposed to Microcystis are poorly described. In the present study, we used two toxic cyanobacteria strains, Aphanizomenon flos-aquae (Aph1395) and Microcystis aeruginosa strain 905 (Ma905) as research subjects. Aph1395 was inhibited with a necessarily concentrated culture filtrate of Ma905 (MA905-SPE), and the response of the inhibited Aph1395 cells was explored via non-targeted metabolomic profiling. In total, 3735 features were significantly different in the Aph1395 treated with Ma905-SPE vs. those treated with BG11 medium. Among them, the annotations of 146 differential features were considered to be confident via MS/MS spectrum matching analysis. Based on the reported physiological functions of the annotated differential features, we proposed a putative model that in the growth-inhibited Aph1395, a suite of increased or decreased features with activities in apoptosis, growth inhibition, and stress response processes contributed to, or defended against, the allelopathic effect caused by Ma905. Our findings provide insights into the interaction between the bloom forming cyanobacterial species that share the same ecological environment.

Widespread Distribution and Adaptive Degradation of Microcystin Degrader (mlr-Genotype) in Lake Taihu, China.

Microbial degradation is an important route for removing environmental microcystins (MCs). Here, we investigated the ecological distribution of microcystin degraders (mlr-genotype), and the relationship between the substrate specificity of the microcystin degrader and the profile of microcystin congener production in the habitat. We showed that microcystin degraders were widely distributed and closely associated with Microcystis abundance in Lake Taihu, China. We characterized an indigenous degrader, Sphingopyxis N5 in the northern Lake Taihu, and it metabolized six microcystin congeners in increasing order (RR > LR > YR > LA > LF and LW). Such a substrate-specificity pattern was congruent to the order of the dominance levels of these congeners in northern Lake Taihu. Furthermore, a meta-analysis on global microcystin degraders revealed that the substrate-specificity patterns varied geographically, but generally matched the profiles of microcystin congener production in the degrader habitats, and the indigenous degrader typically metabolized well the dominant MC congeners, but not the rare congeners in the habitat. This highlighted the phenotypic congruence between microcystin production and degradation in natural environments. We theorize that such congruence resulted from the metabolic adaptation of the indigenous degrader to the local microcystin congeners. Under the nutrient microcystin selection, the degraders might have evolved to better exploit the locally dominant congeners. This study provided the novel insight into the ecological distribution and adaptive degradation of microcystin degraders.

Cyanobacterial blooms in China: diversity, distribution, and cyanotoxins.

Cyanobacterial blooms, which refer to the massive growth of harmful cyanobacteria, have altered the global freshwater ecosystems during the past decades. China has the largest population in the world, and it is suffering from the harmful effect of water eutrophication and cyanobacterial blooms along with rapid development of the economy and society. Research on cyanobacterial blooms and cyanotoxins in China have been overwhelmingly enhanced and emphasized during the past decades. In the present review, the research on cyanobacterial blooms in China is generally introduced, including the history of cyanobacterial bloom studies, the diversity of the bloom-forming cyanobacteria species (BFCS), and cyanotoxin studies in China. Most studies have focused on Microcystis, its blooms, and microcystins. Newly emerging blooms with the dominance of non-Microcystis BFCS have been gradually expanding to wide regions in China. Understanding the basic features of these non-Microcystis BFCS and their blooms, including their diversity, occurrence, physio-ecology, and harmful metabolites, will provide direction on future studies of cyanobacterial blooms in China.

Simultaneous Removal of the Freshwater Bloom-Forming Cyanobacterium Microcystis and Cyanotoxin Microcystins via Combined Use of Algicidal Bacterial Filtrate and the Microcystin-Degrading Enzymatic Agent, MlrA.

Freshwater cyanobacterial blooms (e.g., Microcystis blooms) constitute a major global environmental problem because of their risks to public health and aquatic ecological systems. Current physicochemical treatments of toxic cyanobacteria cause the significant release of cyanotoxin microcystins from damaged cells. Biological control is a promising eco-friendly technology to manage harmful cyanobacteria and cyanotoxins. Here, we demonstrated an efficient biological control strategy at the laboratory scale to simultaneously remove Microcystis and microcystins via the combined use of the algicidal bacterial filtrate and the microcystin-degrading enzymatic agent. The algicidal indigenous bacterium Paenibacillus sp. SJ-73 was isolated from the sediment of northern Lake Taihu, China, and the microcystin-degrading enzymatic agent (MlrA) was prepared via the heterologous expression of the mlrA gene in the indigenous microcystin-degrading bacterium Sphingopyxis sp. HW isolated from Lake Taihu. The single use of a fermentation filtrate (5%, v/v) of Paenibacillus sp. SJ-73 for seven days removed the unicellular Microcystis aeruginosa PCC 7806 and the native colonial Microcystis strain TH1701 in Lake Taihu by 84% and 92%, respectively, whereas the single use of MlrA removed 85% of microcystins. Used in combination, the fermentation filtrate and MlrA removed Microcystis TH1701 and microcystins by 92% and 79%, respectively. The present biological control thus provides an important technical basis for the further development of safe, efficient, and effective measures to manage Microcystis blooms and microcystins in natural waterbodies.

Quantitative Proteomic and Microcystin Production Response of Microcystis aeruginosa to Phosphorus Depletion.

Microcystis blooms are the most widely distributed and frequently occurring cyanobacterial blooms in freshwater. Reducing phosphorus is suggested to be effective in mitigating cyanobacterial blooms, while the underlying molecular mechanisms are yet to be elucidated. In the present study, isobaric tags for relative and absolute quantitation (iTRAQ)-based quantitative proteomics was employed to study the effects of phosphorus depletion on Microcystis aeruginosa FACHB-905. The production of microcystins (MCs), a severe hazard of Microcystis blooms, was also analyzed. In total, 230 proteins were found to be differentially abundant, with 136 downregulated proteins. The results revealed that, upon phosphorus limitation stress, Microcystis aeruginosa FACHB-905 raised the availability of phosphorus primarily by upregulating the expression of orthophosphate transport system proteins, with no alkaline phosphatase producing ability. Phosphorus depletion remarkably inhibited cell growth and the primary metabolic processes of Microcystis, including transcription, translation and photosynthesis, with structures of photosystems remaining intact. Moreover, expression of nitrogen assimilation proteins was downregulated, while proteins involved in carbon catabolism were significantly upregulated, which was considered beneficial for the intracellular balance among carbon, nitrogen and phosphorus. The expression of MC synthetase was not significantly different upon phosphorus depletion, while MC content was significantly suppressed. It is assumed that phosphorus depletion indirectly regulates the production of MC by the inhibition of metabolic processes and energy production. These results contribute to further understanding of the influence mechanisms of phosphorus depletion on both biological processes and MC production in Microcystis cells.

The involvement of α-proteobacteria Phenylobacterium in maintaining the dominance of toxic Microcystis blooms in Lake Taihu, China.

Lake Taihu in China has suffered serious harmful cyanobacterial blooms for decades. The algal blooms threaten the ecological sustainability, drinking water safety, and human health. Although the roles of abiotic factors (such as water temperature and nutrient loading) in promoting Microcystis blooms have been well studied, the importance of biotic factors (e.g. bacterial community) in promoting and meditating Microcystis blooms remains unclear. In this study, we investigated the ecological dynamics of bacterial community, the ratio of toxic Microcystis, as well as microcystin in Lake Taihu. High-throughput 16S rRNA sequencing and principal component analysis (PCA) revealed that the bacteria community compositions (BCCs) clustered into three groups, the partitioning of which corresponded to that of groups according to the toxic profiles (the ratio of toxic Microcystis to total Microcystis, and the microcystin concentrations) of the samples. Further Spearman's correlation network showed that the α-proteobacteria Phenylobacterium strongly positively correlated with the toxic profiles. Subsequent laboratory chemostats experiments demonstrated that three Phenylobacterium strains promoted the dominance of the toxic Microcystis aeruginosa PCC7806 when co-culturing with the non-toxic PCC7806 mcyB- mutant. Taken together, our data suggested that the α-proteobacteria Phenylobacterium may play a vital role in the maintenance of toxic Microcystis dominance in Lake Taihu.

Understanding the Differences in the Growth and Toxin Production of Anatoxin-Producing Cuspidothrix issatschenkoi Cultured with Inorganic and Organic N Sources from a New Perspective: Carbon/Nitrogen Metabolic Balance.

Cyanotoxins are the underlying cause of the threat that globally pervasive Cyanobacteria Harmful algal blooms (CyanoHABs) pose to humans. Major attention has been focused on the cyanobacterial hepatotoxin microcystins (MCs); however, there is a dearth of studies on cyanobacterial neurotoxin anatoxins. In this study, we explored how an anatoxin-producing Cuspidothrix issatschenkoi strain responded to culture with inorganic and organic nitrogen sources in terms of growth and anatoxins production. The results of our study revealed that ʟ- alanine could greatly boost cell growth, and was associated with the highest cell productivity, while urea significantly stimulated anatoxin production with the maximum anatoxin yield reaching 25.86 µg/mg dry weight, which was 1.56-fold higher than that in the control group (BG11). To further understand whether the carbon/nitrogen balance in C. issatschenkoi would affect anatoxin production, we explored growth and toxin production in response to different carbon/nitrogen ratios (C/N). Anatoxin production was mildly promoted when the C/N ratio was within low range, and significantly inhibited when the C/N ratio was within high range, showing approximately a three-fold difference. Furthermore, the transcriptional profile revealed that anaC gene expression was significantly up-regulated over 2-24 h when the C/N ratio was increased, and was significantly down-regulated after 96 h. Overall, our results further enriched the evidence that urea can stimulate cyanotoxin production, and ʟ-alanine could boost C. issatschenkoi proliferation, thus providing information for better management of aquatic systems. Moreover, by focusing on the intracellular C/N metabolic balance, this study explained the anatoxin production dynamics in C. issatschenkoi in response to different N sources.

The Proposed Neurotoxin ß-N-Methylamino-l-Alanine (BMAA) Is Taken up through Amino-Acid Transport Systems in the Cyanobacterium Anabaena PCC 7120.

Produced by cyanobacteria and some plants, BMAA is considered as an important environmental factor in the occurrence of some neurodegenerative diseases. Neither the underlying mechanism of its toxicity, nor its biosynthetic or metabolic pathway in cyanobacteria is understood. Interestingly, BMAA is found to be toxic to some cyanobacteria, making it possible to dissect the mechanism of BMAA metabolism by genetic approaches using these organisms. In this study, we used the cyanobacterium Anabaena PCC 7120 to isolate BMAA-resistant mutants. Following genomic sequencing, several mutations were mapped to two genes involved in amino acids transport, suggesting that BMAA was taken up through amino acid transporters. This conclusion was supported by the protective effect of several amino acids against BMAA toxicity. Furthermore, targeted inactivation of genes encoding different amino acid transport pathways conferred various levels of resistance to BMAA. One mutant inactivating all three major amino acid transport systems could no longer take up BMAA and gained full resistance to BMAA toxicity. Therefore, BMAA is a substrate of amino acid transporters, and cyanobacteria are interesting models for genetic analysis of BMAA transport and metabolism.

Microcystin-LR Degradation and Gene Regulation of Microcystin-Degrading Novosphingobium sp. THN1 at Different Carbon Concentrations.

The bacterium Novosphingobium sp. THN1 (THN1) is capable of degrading microcystin-LR (MC-LR). To study the ability of THN1 to degrade MC-LR and its possible mechanism(s) of regulation, we analyzed the effect of carbon concentrations on the degradation process. The MC-LR degradation rate peaked early and then declined during MC-LR biodegradation. Decreased levels of carbon in the medium caused the degradation peak to occur earlier. The expression of the functional gene mlrA, encoding a microcystinase, showed a similar trend to the MC-LR degradation rate at various carbon concentrations (r2 = 0.717, p < 0.05), suggesting that regulation of mlrA expression may play an important role in MC-LR degradation by THN1. The total bacterial biomass decreased when the carbon source was limited and did not correlate with the MC-LR degradation rate. Transcriptomic analysis showed that MC-LR degradation differentially regulated 62.16% (2597/4178) of THN1 genes. A considerable number of differentially expressed genes (DEGs) during MC-LR degradation encoded proteins related to carbon-, nitrogen-, and amino acid-related pathways. At 2 h of MC-LR degradation, most DEGs (29/33) involved in carbon and nitrogen metabolism were downregulated. This indicated that MC-LR may regulate carbon and nitrogen pathways of Novosphingobium sp. THN1. KEGG pathway analysis indicated that the upregulated DEGs during MC-LR degradation were mainly related to amino acid degradation and substrate metabolism pathways. Particularly, we detected increased expression of glutathione metabolism-related genes from transcriptomic data at 2 h of MC-LR degradation compared with the gene expression of 0 h, such as GST family protein, glutathione peroxidase, S-(hydroxymethyl) glutathione dehydrogenase, and glutathione-dependent disulfide-bond oxidoreductase that have been reported to be involved in microcystin degradation.

The highly heterogeneous methylated genomes and diverse restriction-modification systems of bloom-forming Microcystis.

The occurrence of harmful Microcystis blooms is increasing in frequency in a myriad of freshwater ecosystems. Despite considerable research pertaining to the cause and nature of these blooms, the molecular mechanisms behind the cosmopolitan distribution and phenotypic diversity in Microcystis are still unclear. We compared the patterns and extent of DNA methylation in three strains of Microcystis, PCC 7806SL, NIES-2549 and FACHB-1757, using Single Molecule Real-Time (SMRT) sequencing technology. Intact restriction-modification (R-M) systems were identified from the genomes of these strains, and from two previously sequenced strains of Microcystis, NIES-843 and TAIHU98. A large number of methylation motifs and R-M genes were identified in these strains, which differ substantially among different strains. Of the 35 motifs identified, eighteen had not previously been reported. Strain NIES-843 contains a larger number of total putative methyltransferase genes than have been reported previously from any bacterial genome. Genomic comparisons reveal that methyltransferases (some partial) may have been acquired from the environment through horizontal gene transfer.

Microcystin-Bound Protein Patterns in Different Cultures of Microcystis aeruginosa and Field Samples.

Micocystin (MC) exists in Microcystis cells in two different forms, free and protein-bound. We examined the dynamic change in extracellular free MCs, intracellular free MCs and protein-bound MCs in both batch cultures and semi-continuous cultures, using high performance liquid chromatography and Western blot. The results showed that the free MC per cell remained constant, while the quantity of protein-bound MCs increased with the growth of Microcystis cells in both kinds of culture. Significant changes in the dominant MC-bound proteins occurred in the late exponential growth phase of batch cultures, while the dominant MC-bound proteins in semi-continuous cultures remained the same. In field samples collected at different months in Lake Taihu, the dominant MC-bound proteins were shown to be similar, but the amount of protein-bound MC varied and correlated with the intracellular MC content. We identified MC-bound proteins by two-dimensional electrophoresis immunoblots and mass spectrometry. The 60 kDa chaperonin GroEL was a prominent MC-bound protein. Three essential glycolytic enzymes and ATP synthase alpha subunit were also major targets of MC-binding, which might contribute to sustained growth in semi-continuous culture. Our results indicate that protein-bound MC may be important for sustaining growth and adaptation of Microcystis sp.

Microcystin-degrading bacteria affect mcyD expression and microcystin synthesis in Microcystis spp.

Cyanobacterial blooms occur increasingly often and cause ecological, economic and human health problems worldwide. Microcystins (MCs) are the dominant toxins produced by cyanobacteria and are implicated in epidemic disease and environmental problems. Extensive research has been reported on the various regulating factors, e.g., light, temperature, nutrients such as nitrogen and phosphorus, pH, iron, xenobiotics, and predators, that influence microcystin (MC) synthesis, but little is known about the effects of cyanobacteria-associated bacteria on MC synthesis. A considerable number of studies have focused on interactions between Microcystis species and their associated bacteria. In this study, we evaluated the effects of MC-degrading bacteria (MCDB) on MC synthesis gene mcyD expression and MC synthesis in axenic strain PCC7806, non-axenic strain FACHB905, and colony strain FACHB1325 of Microcystis by quantitative real-time polymerase chain reaction (RT-PCR) assay and enzyme-linked immunosorbent assay (ELISA). We demonstrate for the first time that MCDB can induce and up-regulate the MC production and transcriptional response of the mcyD gene of toxic Microcystis. On day 4 of the culturing experiment, the intracellular MC concentration and transcriptional response of mcyD of FACHB1325 were up-regulated 1.9 and 5.3-fold over that of the control, and for FACHB905 were up-regulated 1.8 and 4.2-fold over that of the control, respectively. On day 10, the transcriptional response of mcyD was up-regulated 21.3-fold in PCC7806. These results indicate that there are interactions between toxic Microcystis and MCDB, and MCDB may play a role in regulating mcyD expression in toxic Microcystis.

Multi-Year Assessment of Toxic Genotypes and Microcystin Concentration in Northern Lake Taihu, China.

Lake Taihu is the third-largest freshwater lake in China and has been suffering from cyanobacterial blooms for over two decades. The northern part of the lake, Meiliang Bay, is known to be at high risk of dense and sustained Microcystis blooms and toxins. This study aimed to investigate and record the annual and seasonal dynamics of toxic genotype, Microcystis morphospecies succession and microcystin variation. It also aimed to find out the underlying driving factors influencing the dynamic changes. Microcystin (MC) and the Microcystis genotype were quantified using HPLC and quantitative real-time PCR, respectively. Our study, over three consecutive years, showed that the pattern of morphospecies succession was seasonally distinct and annually consistent. During the same period in 2012, 2013 and 2014, the average MC were, on dry weight basis, 733 µg·g(-1), 844 µg·g(-1), 870 µg·g(-1), respectively. The proportion of toxic Microcystis accounted for 41%, 44% and 52%, respectively. Cell bound microcystin was found to correlate with the percentage of toxic Microcystis. Based on historical and current data, we conclude that annual bloom toxicity was relatively stable or possibly increased over the last decade.

Harmful algal bloom removal and eutrophic water remediation by commercial nontoxic polyamine-co-polymeric ferric sulfate-modified soils.

Harmful algal bloom has posed great threat to drinking water safety worldwide. In this study, soils were combined with commercial nontoxic polyamine poly(epichlorohydrin-dimethylamine) (PN) and polymeric ferric sulfate (PFS) to obtain PN-PFS soils for Microcystis removal and eutrophic water remediation under static laboratory conditions. High pH and temperature in water could enhance the function of PN-PFS soil. Algal removal efficiency increased as soil particle size decreased or modified soil dose increased. Other pollutants or chemicals (such as C, P, and organic matter) in eutrophic water could participate and promote algal removal by PN-PFS soil; these pollutants were also flocculated. During PN-PFS soil application in blooming field samples, the removal efficiency of blooming Microcystis cells exceeded 99 %, the cyanotoxin microcystins reduced by 57 %. Water parameters (as TP, TN, SS, and SPC) decreased by about 90 %. CODMn, PO4-P, and NH4-N also sharply decreased by >45 %. DO and ORP in water improved. Netting and bridging effects through electrostatic attraction and complexation reaction could be the two key mechanisms of Microcystis flocculation and pollutant purification. Considering the low cost of PN-PFS soil and its nontoxic effect on the environment, we proposed that this soil combination could be applied to remove cyanobacterial bloom and remediate eutrophic water in fields.

Distribution and population dynamics of potential anatoxin-a-producing cyanobacteria in Lake Dianchi, China.

The occurrence of cyanobacterial blooms is often accompanied by a variety of toxic secondary metabolites known as cyanotoxins. Anatoxin-a (ATX-a) is a highly toxic cyanobacterial neurotoxin synthesized by numerous species (e.g., Aphanizomenon, Anabaena and Oscillatoria) that has received much public attention. In this study, we used molecular methods (PCR and qPCR) to track the presence and dynamics of ATX-a-producing cyanobacteria, Aphanizomenon and Anabaena in Lake Dianchi, China based on the anaC and cpcBA-IGS genes over a 23-month period (from June 2010 to April 2012). Results revealed that Aphanizomenon was the major potential ATX-a producer in Lake Dianchi and that they were most abundant in early spring and least abundant in summer, coinciding with observed Aphanizomenon blooms. It was found that the proportion of ATX-a toxigenic cells was lower in the northern part of the lake (2.1%) than the middle (16.7%) and southern parts (19.2%). The information on the spatio-temporal distributions of ATX-a-producing cyanobacteria obtained in this study will help to build management strategies to improve water quality for public health.

Seasonal dynamics of water bloom-forming Microcystis morphospecies and the associated extracellular microcystin concentrations in large, shallow, eutrophic Dianchi Lake.

The increasing occurrence of Microcystis blooms is of great concern to public health and ecosystem due to the potential hepatotoxic microcystins (MCs) produced by these colonial cyanobacteria. In order to interpret the relationships between variations of Microcystis morphospecies and extracellular MC concentrations, the seasonal dynamics of phytoplankton community composition, MC concentrations, and environmental parameters were monitored monthly from August, 2009 to July, 2010. The results indicated that Microcystis dominated total phytoplankton abundance from May to December (96%-99% of total biovolume), with toxic Microcystis viridis and non-toxic Microcystis wesenbergii dominating after July (constituting 65%-95% of the Microcystis population), followed by M. viridis as the sole dominant species from November to January (49%-93%). Correlation analysis revealed that water temperature and nutrient were the most important variables accounting for the occurrence of M. wesenbergii, while the dominance of M. viridis was related with nitrite and nitrate. The relatively low content of MCs was explained by the association with a large proportion of M. viridis and M. wesenbergii, small colony size of Microcystis populations, and low water temperature, pH and dissolved oxygen. The extracellular MC (mean of 0.5±0.2µg/L) of water samples analyzed by enzyme-linked immunosorbent assay (ELISA) demonstrated the low concentrations of MC in Dianchi Lake which implied the low potential risk for human health in the basin. The survey provides the first whole lake study of the occurrence and seasonal variability of Microcystis population and extracellular MCs that are of particular interest for water quality monitoring and management.