Cultivation and Characterization of the MaMV-DC Cyanophage that Infects Bloom-forming Cyanobacterium Microcystis aeruginosa

The MaMV-DC cyanophage,which infects the bloom-forming cyanobacterium Microcystis aeruginosa,was isolated from Lake Dianchi,Kunming,China.Twenty-one cyanobacterial strains were used to detect the host range of MaMV-DC.Microcystic aeruginosa FACHB-524 and plaque purification were used to isolate individual cyanophages,and culturing MaMV-DC with cyanobacteria allowed us to prepare purified cyanophages for further analysis.Electron microscopy demonstrated that the negatively stained viral particles are tadpole-shaped with an icosahedral head approximately 70 nm in diameter and a contractile tail approximately 160 nm in length.Using one-step growth experiments,the latent period and burst size of MaMV-DC were estimated to be 24–48 hours and approximately 80infectious units per cell,respectively.Restriction endonuclease digestion and agarose gel electrophoresis were performed using purified MaMV-DC genomic DNA,and the genome size was estimated to be approximately 160 kb.Sodium dodecyl sulfate polyacrylamide gel electrophoresis(SDS-PAGE)analysis revealed four major structural proteins.These results support the growing interest in using freshwater cyanophages to control bloom-forming cyanobacterium.

The trophic state of lake water regulates spatial-temporal variations of bloom-forming Microcystis

Although biomass variations in Microcystis and microcystin have been widely reported,few studies have addressed whether different trophic states of natural lake water affect the spatial-temporal variations in abundances of microcystin-producing Microcystis in a given bloom.In this study,we used a harmful algal bloom in Chaohu Lake,eastern China,as an example to investigate the mutual relationship between different nutrient states and environmental factors,and the impact on Microcystis.Overall,cyanobacteria and Microcystis were more abundant in the middle and western parts of the lake under high nutrients levels,while in the eastern part,nutrient concentrations were low enough to limit biomass,and their fluctuations affected the contents of toxic Microcystis.Moreover,microcystin concentration was correlated positively to nutrient levels and Microcystis biomass during bloom developing in 2013 from June to August.Temporally,the cellular content of total microcystin was lowest when the bloom peaked in intensity.Our results suggest that lake eutrophication not only results in cyanobacterial blooms,but may also increase the proportion of toxic Microcystis species and their cell-bound MCs contents(i.e.microcystin cell quotas) under mild eutrophication.The present investigation provided molecular evidence for the selection of MC-producing and non-MC-producing genotypes.The current study provides new evidence advocating the monitoring of partitions of large lakes when studying cyanobacteria and toxin-contaminated freshwaters,which will be beneficial for both water agencies and water researchers.

Role of Suspended Sediments and Mixing in Reducing Photoinhibition in the Bloom-Forming Cyanobacterium Microcystis

Toxic cyanobacterial blooms are becoming a global problem. Previous research of cyanobacterial bloom development has examined how high nutrient concentrations promote cyanobacteria dominance, and how positive buoyancy provides an ecological advantage over sinking phytoplankton. Tributaries responsible for loading nutrients into lakes often simultaneously contribute high concentrations of suspended sediments. High concentrations of suspended sediments may also influence blooms by affecting the ambient light climate, reducing photodamage, and increasing the efficiency of photosynthesis. We examined the effects of sediments and vertical mixing in potentially reducing photodamage to Microcystis by measuring photosynthetic parameters and pigment content of Microcystis in western Lake Erie during the 2008 bloom and in laboratory experiments. Photosynthetic efficiency increased with increasing sediment concentration in the lake and laboratory experiment. Content of photo-protective carotenoid pigments per dry weight decreased with increasing sediment concentrations, while the light-harvesting pigments, chl a and phycocyanin, increased with sediments. These results indicate that suspended sediments reduce photoinhibition for Microcystis. Further, photosynthetic damage was higher when Microcystis was concentrated on the surface compared to a mixed water column. Measurements of Microcystis abundance and light were also recorded, in addition to photosynthetic measurements. Greatest Microcystis abundances in Lake Erie were recorded during light-limiting conditions, which offer Microcystis both physiological and ecological benefits by reducing photoinhibition and increasing Microcystis’ advantage in light competition via buoyancy. Efforts to reduce cyanobacterial blooms may include reducing suspended sediments loads in combination with reducing nutrient loading.

Periodic Addition of Glucose Suppressed Cyanobacterial Abundance in Additive Lake Water Samples during the Entire Bloom Season

Previously, we showed that prophylactic addition of glucose to Harsha Lake water samples could inhibit cyanobacteria growth, at least for a short period of time. The current study tested cyanobacterial control with glucose for the entire Harsha Lake bloom season. Water samples (1000 ml) were collected weekly from Harsha Lake during the algal-bloom season starting June 9 and lasting until August 24, 2022. To each of two 7-liter polypropylene containers, 500 ml of Harsha Lake water was added, and the containers were placed in a controlled environment chamber. To one container labeled “Treated,” 0.15 g of glucose was added, and nothing was added to the container labeled “Control.” After that, three 25 ml samples from each container were collected and used for 16S rRNA gene sequencing each week. Then 1000 ml of Harsha Lake water was newly collected each week, with 500 ml added to each container, along with the addition of 0.15 g glucose to the “Treated” container. Sequencing data were used to examine differences in the composition of bacterial communities between Treated and Control containers. Treatment with glucose altered the microbial communities by 1) reducing taxonomic diversity, 2) largely eliminating cyanobacterial taxa, and 3) increasing the relative abundance of subsets of non-cyanobacterial taxa (such as Proteobacteria and Actinobacteriota). These effects were observed across time despite weekly inputs derived directly from Lake water. The addition of glucose to a container receiving weekly additions of Lake water suppressed the cyanobacterial populations during the entire summer bloom season. The glucose appears to stimulate the diversity of certain bacterial taxa at the expense of the cyanobacteria.

Ultra-high specific surface area activated carbon from Taihu cyanobacteria via KOH activation for enhanced methylene blue adsorption

Cyanobacteria-based activated carbon(CBAC)was successfully prepared by pyrolysis-activation of Taihu cyanobacteria.When the impregnation ratio and activated temperature were 2 and 800-C,respectively,the optimal CBACs possessed an ultra-high specific surface(2178.90 m^(2)·g^(-1))and plenty of micro-and meso-pores,as well as a high pore volume(1.01 cm^(3)·g^(-1)).Ascribed to ultra-high surface area,π-π interaction,electrostatic interaction,as well as hydrogen-bonding interactions,the CBACs displayed huge superiority in efficient dye removal.The saturated methylene blue adsorption capacity by CBACs could be as high as 1143.4 mg·g^(-1),superior to that of other reported biomass-activated carbons.The adsorption was endothermic and modeled well by the pseudo-second-order kinetic,intra-particle diffusion,and Langmuir models.This work presented the effectiveness of Taihu cyanobacteria adsorbent ascribed to its super large specific surface area and high adsorption ability.

Microscopic analysis on eukaryotic algae and cyanobacteria in nine seasonal lakes and ponds in Vestfjella,Dronning Maud Land,Antarctica

Antarctic continental lakes and ponds are among the most impoverished aquatic environments on earth but many of them support flourishing populations of cyanobacteria,eukaryotic algae,protozoans,and some multicellular animals.In this study,we present results of a microscopic analysis of cyanobacteria and eukaryotic algae from nine diverse types of Antarctic continental water bodies during one austral summer.The results supplement and enlarge our previous studies on the limnological characteristics of the epiglacial and supraglacial lakes and ponds in Dronning Maud Land,an area that has received little attention from limnologists.The taxon with highest frequency among the samples(n=79)was Mesotaenium cf.berggrenii,a eukaryotic Zygnematophyceae,which occurred in 82%of the samples with a maximum cell density of 68 cells·mL^(-1).The taxa with second and third highest frequency were the prokaryotes Gloeocapsopsis(60%)and Leptolyngbya(41%),followed by Chlamydomonas(34%)and Cyanothece(29%).The number of taxa varied between 7-21 among the lakes and ponds,being highest in a supraglacial lake,and lowest in an epiglacial lake.The results did not reveal any obvious correlation between the abundance of any taxa and the water chemistry,but water bodies with inorganic sediments had higher cell densities and biomasses than those without sediment.This suggests the importance of sediment in supporting biological diversity in these ultraoligotrophic lakes and ponds.

Measurement of Gas Vesicle in Cyanobacteria and Its Pretreatment Methods

[Objective]The aim was to measure gas vesicle in cyanobacteria and discuss its pretreatment methods.[Method]The capillary pressure method to determine gas vesicle in cyanobacteria was modified firstly,and then the lower detection limit and precision of modified apparatus were tested,finally the effects of two concentration methods and preservation methods on cell quantity and gas vesicle content of three cyanobacterias were studied.[Result]The lower detection limit and precision of modified apparatus to measure gas vesicle in three cyanobacterias were 0.001 8 μl/ml and 1% respectively.Unicellular Microcystis couldn＇t be concentrated effectively by filtration or centrifugation method,and the loss rate reached 50%.However,the colony of Microcystis and filamentous Planktothrix mougeotii could be effectively concentrated by centrifugation and filtration method respectively,with low loss rate.Besides,the effects of filtration and centrifugation on the content of gas vesicle in cells could be neglected.After preserved by direct refrigeration and adding Lugol＇s iodine solution for 7 d,there was no obvious change in cell concentration and gas vesicle content per cell,and the loss of gas vesicle under direct refrigeration was small,while the preservation of natural water samples should add Lugol＇s iodine solution.[Conclusion]The study could provide theoretical foundation for the researches on buoyancy regulation mechanism and blooming mechanism of cyanobacteria.

宁波沿海陆源排污口蓝藻(Cyanobacteria)和裸藻纲(Euglenoidea)生物的分布特点

采用高通量454焦磷酸测序方法对宁波沿海2个重点排污口、8个一般排污口的20个站位水样进行分析,得到2011年的3月、5月、8月和10月份各排污口蓝藻和含叶绿体单细胞生物的分布情况。研究结果表明:宁波沿海陆源排污口中含叶绿体单细胞生物数量最多,占91.857%,包括眼虫门(Euglenozoa)、绿藻门(Chlorophyta)、隐藻门(Cryptophyta)生物。其中主要为眼虫门的裸藻纲(Euglenoidea)生物占90.08%。蓝藻门的集球藻目(Synechococcophycideae)和颤藻亚目(Oscillatoriophycideae)含量相对较多,分别占5.70%和1.79%;排污口中裸藻纲生物数量季节性差异比较大,3月份、8月份数量相对较多,5月份和10月份较少,推测这类生物的数量与季节性温度变化有关;各排污口污染物、这两类生物检出量和种类有一定差异,在氮磷含量高的排污口(如S9号)其检出量并不高,推测检出量与排污口的主要污染物中氨氮含量没有直接关系,可能受氮磷比的影响。

Research process of the calcium signaling in cyanobacteria

The potential involvement of calcium in signalling in cyanobacteria has been investigated in recent years. Enough evidences showed that the cyanobacteria were capable of sensing and distinguishing different environmental stimuli, and making responses in ways of Ca^2＋ transients, which were the results of influx or efflux of Ca^2＋ aroused by different environmental stimuli. The calcium signal elicited by nitrogen starvation was crucial to heterocyst differentiation in filamentous cyanobacteria Anabaena species. Identification of a calcium-binding protein （CcbP） from Anabaena sp. PCC 7120 provided further evidence, and the degradation and down-regulation of CcbP accounted for the generation of calcium signal when nitrogen starvation exits. However, the encoding and decoding mechanisms of the calcium signals in cyanobacteria still remain unclear. In order to reveal the exact role of it, a detailed, systematic investigation will be needed, especially for the calcium dynamics at the single cell level.

The morphological and molecular detection for the presence of toxic Cylindrospermopsis(Nostocales, Cyanobacteria) in Beijing city, China

Cylindrospermopsis raciborskii and its highly similar relatives Raphidiopsis species have been recognized as globally invasive and expansive filamentous cyanobacteria causing water blooms. Reports on C. raciborsMi/Raphidiopsis species and their harmful metabolites such as hepatotoxic cylindrospermopsins （CYNs） in Chinese waters have been increasing, but mostly restricted to the southern regions of China. To further explore the existence and distribution of C. raciborskii in China, six water samples from Beijing city were morphologically and molecularly examined. Five samples of the six were shown to have Cylindrospermopsis filaments with straight and spiral morphotypes. PCR detection targeting on Cylindrospermopsis/Raphidiopsis specific 16S rRNA gene region also showed the positive amplification, and such amplifications were confirmed by sequencing and phylogenetic analysis. As well, three of the five Cylindrospermopsis containing samples were shown to have cyrJ--a gene of CYN synthesis gene duster. The results represented the presence of toxic Cylindrospermopsis at the most northern line in China so far, indicating rapid expansion of this harmful invasive cyanobacterium. It is strongly suggested that the monitoring on C. raciborskii/Raphidiopsis species and their production of cylindrospermopsin should be emphasized in Beijing and even more northem parts of China.

Fractional pyrolysis of Cyanobacteria from water blooms over HZSM-5 for high quality bio-oil production

Fractional pyrolysis and one-step pyrolysis of natural algae Cyanobacteria from Taihu Lake were comparatively studied from 200 to 500 ℃. One-step pyrolysis produced bio-oil with complex composition and low high heating value （HHV〈30.9 MJ/kg）. Fractional pyrolysis separated the degradation of different components in Cyanobacteria and improved the selectivity to products in bio-oil. That is, acids at 200 ℃, amides and acids at 300 ℃, phenols and nitriles at 400 ℃, and phenols at 500 ℃, were got as main products, respectively. HZSM-5 could promote the dehydration, cracking and aromatization of pyrolytic intermediates in fractional pyrolysis. At optimal HZSM-5 catalyst dosage of 1.0 g, the selectivity to products and the quality of bio-oil were improved obviously. The main products in bio-oil changed to nitriles （47.2%） at 300 ℃, indoles （51.3%） and phenols （36.3%） at 400 ℃. The oxygen content was reduced to 7.2 wt% and 9.4 wt%, and the HHV was raised to 38.1 and 37.3 MJ/kg at 300 and 400 ℃, respectively. Fractional catalytic pyrolysis was proposed to be an efficient method not only to provide a potential solution for alleviating environmental pressure from water blooms, but also to improve the selectivity to products and obtain high quality bio-oil.

Bacterial and cyanobacterial diversities determined by T-RFLP analyses in the Jiaozhou Bay

The methods of DAPI staining epifluorescence microscopy and T-RFLP analysis were used to analyze the microbial abundance and diversity in surface seawater sampled from 12 stations inside and outside of the Jiaozhou Bay during a survey on 12 and 13 September 2004. The abundance of total microbes is in the range of 10^6～ 10^7 cells/cm^3, similar to those of most semi-enclosed bays in the temperate areas in the world. The highest microbial densities occur in the northeastern part of the Jiaozhou Bay, around the mouths of Loushan and Licun Rivers and the Hongdao aquacultural farming areas, suggesting that the degree and characteristics of pollutions, along with geographical and hydrological effects, may be important determinants affecting the abundance and distribution of bacteria in the Jiaozhou Bay. Bacterial communities inside and outside of the Jiaozhou Bay can be grouped into three classes based on T-RFLP patterns and cluster analyses. Stations at the water channel of the bay mouth and outside, such as D1, D3, D5, D6 and D7, are grouped together to stand for the outside bacterial community interacting with the environment outside of the Jiaozhou Bay. Stations of the innermost side of the Jiaozhou Bay, such as A3, A5, B2 and Y1, are grouped together to stand for the residential bacteria community. Stations C1, C3 and CA are grouped together and may stand for the transitional bacterial assemblage between the residential community and the outside community. However, there is no such a defined relationship for the case of cyanobacterial diversity, indicating the fact that cyanobacteria are more flexible and adaptable to all kinds of conditions.

Removal of heavy metals from mine water by cyanobacterial calcification

The influence of different illumination intensities on cyanobacterial calcification induced removal of heavy metals from contaminated mine water was studied. Cyanobacterial calcification experiments were performed using a growth medium intended to simulate contaminated mine water. The results indicate that calcification can promote the removal of heavy metal ions. As the illumination intensity became stronger calcification rates increased and the removal of Zn2+ and Cd2+ became more obvious. When the illumination intensity was 10000 lux the removal of Pb2+ was the largest observed: stronger or weaker illumination reduced the amount of lead removed. The removal of three different heavy metals complies with an index function. For identical illumination intensities different ions were removed to different degrees.

Variation of phytoplankton community before an induced cyanobacterial(Arthrospira platensis) bloom

A cyanobacterial （Arthrospira platensis） bloom was induced in situ by nutrient manipulation in an enclosure. The succession of the phytoplankton community and the water chemistry variations before the appearance of bloom, as well as their relationship, were investigated. The cell pigment variations were studied simultaneously. The Pearson＇s correlation analysis showed that there was no significant correlation between water chemistry and green algal or cyanobacterial composition, indicating that water chemistry variations were not suitable to be used as indicators for cyanobacterial-bloom early-warning. However, the diversity index of the phytoplankton community decreased sharply before the bloom appeared. Therefore, the dynamics of phytoplankton community was put forward to be an indicator for cyanobacterial-bloom early-warning. In addition, the cell pigment variations represented the changes of community structure, which should be useful for studying the dynamics of phytoplankton community.

Community Structures of Phytoplankton with Emphasis on Toxic Cyanobacteria in an Ohio Inland Lake during Bloom Season

The community structures of phytoplankton are important factors and indicators of lake water quality. Harmful algal blooms severely impact water supply, recreational activities and wildlife habitat. This study aimed to examine the phytoplankton composition and variations using microscopy, and identify harmful Cyanobacteria in weekly samples taken from four sites at Harsha Lake in southwest Ohio. Over the course of the summer in 2015, the phytoplankton of Harsha Lake consisted mainly of 13 taxa belonging to Bacillariophyta, Chlorophyta, Cryptophyta, Cyanobacteria, Dinophyta and Euglenophyta. Their significant successions started with Bacillariophyta and/or Chlorophyta, then bloomed with Cyanobacteria and ended with Chlorophyta and/or Dinophyta. Cyanobacteria members: Microcystis, Planktothrix, Dolichospermum, Aphanizomenon, Cylindrospermopsis, and Oscillatoria from the Cyanophyceae were identified to be dominant genera. These organisms varied spatially and temporally in similar patterns along with the variations of nutrients and formed the summer bloom with the total biomasses ranging from 0.01 to 114.89 mg L-1 with mean of 22.88 mg L-1. M. aeruginosa and P. rubescens were revealed as the microcystin producers, while A. circinalis and Aphanizomenon sp. were identified as a saxitoxin producer through cloning and sequencing PCR products of mcyA, mcyE and sxtA genes. The biomasses of phytoplankton, Cyanobacteria and Microcystis were positively correlated to nutrients, especially to total nitrogen. The total ELISA measurement for microcystin positively correlated with Cyanobacteria (R2 = 0.66, P Microcystis (R2 = 0.64, P 2 = 0.59, P < 0.0001). The basic information on the occurrence and biomasses of Cyanobacteria and total phytoplankton, and the analysis for toxic species, which were the first report for the inland water in Ohio, USA, will document the succession patterns of phytoplankton and toxin production over a season and provide data to predict risk occurrence to both human and ecological factors.

Hepatitis B virus x gene and cyanobacterial toxins promote aflatoxin B_1-induced hepatotumorigenesis in mice

AIM： To assess the combinative role of aflatoxin B1 （AFB1）, cyanobacterial toxins （cyanotoxins）, and hepatitis B virus （HBV） x gene in hepatotumorigenicity. METHODS： One-week-old animals carrying HBV x gene and their wild-type littermates were intraperitoneally （ip） injected with either single-dose AFB1 [6 mg/kg body weight （bw）], repeated-dose cyanotoxins （microcystin- LR or nodularin, 10 μg/kg bw once a week for 15 wk）, DMSO （vehicle control） alone, or AFB1 followed by cyanotoxins a week later, and were sacrificed at 24 and 52 wk post-treatment. RESULTS： AFB1 induced liver tumors in 13 of 29 （44.8%） transcjenic mice at 52 wk post-treatment, significantly more frequent than in wild-type mice （13.3%）. This significant difference was not shown in the 24-wk study. Compared with AFB1 exposure alone, MC-LR and nodularin yielded approximately 3-fold and 6-fold increases in the incidence of AFB1-induced liver tumors in wild-type animals at 24 wk, respectively. HBV x gene did not further elevate the risk associated with coexposure to AFB1 and cyanotoxins. With the exception of an MC-LR-dosed wild-type mouse, no liver tumor was observed in mice treated with cyanotoxins alone at 24 wk. Neither DMSO-treated transgenic mice nor their wild-type littermates had pathologic alterations relevant to hepatotumorigenesis in even up to 52 wk. CONCLUSION： HBV x gene and nodularin promote the development of AFB1-induced liver tumors. Co-exposure to AFB1 and MC-LR tends to elevate the risk of liver tumors at 24 wk relative to exposure to one of them. The combinative effect of AFB1, cyanotoxins and HBVx on hepatotumorigenesis is weak at 24 wk.

Alternative Methods for Analysis of Cyanobacterial Populations in Drinking Water Supplies: Fluorometric and Toxicological Applications Using Phycocyanin

The management of cyanobacteria and potential exposure to associated biotoxins requires the allocation of scarce resources across a range of freshwater resources within various jurisdictions. Cost effective and reliable methods for sample processing and analysis form the foundation of the protocol yielding reliable data from which to derive important decisions. In this study the utilization of new methods to collect, process and analyze samples enhanced our ability to evaluate cyanobacterial populations. Extraction of phycocyanin using the single freeze thaw method provided more accurate and precise measurements (CV 4.7% and 6.4%), offering a simple and cost-effective means to overcome the influence of morphological variability. In-vacuo concentration of samples prior to ELISA analysis provided a detection limit of 0.001 μg·L?1 MC. Fractionation of samples (?1) = ?0.279 + (1.368 ? Log PC (μg·L?1) while in an Aphanizomemon spp. dominant system Log MC (ng·L?1) = 0.385 + (0.449 ? Log PC (μg·L?1). These methods and sampling protocol could be used in other aquatic systems across a broader regional landscape to estimate the levels of microcystins.

Interactive Effects of Temperature, Nitrogen, and Zooplankton on Growth and Protein and Carbohydrate Content of Cyanobacteria from Western Lake Erie

Harmful algal blooms (HABs) in freshwater ecosystems, especially of cyanobacterial species, are becoming more frequent and expanding geographically, including in Lake Erie in North America. HABs are the result of complex and synergistic environmental factors, though N or P eutrophication is a leading cause. With global mean temperatures expected to increase an additional 2°C - 5°C by 2100, cyanobacterial blooms are predicted to increase even more, given their typically-high temperature optimum for growth. We investigated how increases in temperature and nitrogen, singly or in combination, affect the growth, food quality, and herbivory of Lake Erie cyanobacteria. Algal community samples collected from Lake Erie, and isolated non-N-fixing (Microcystis aeruginosa) and N-fixing (Anabaena flos-aquae) cyanobacterial species, were cultured at 20°C, 25°C, or 30°C, and at 5, 50, 150, or 250 μM N, and then analyzed for growth and (for isolates) content of total protein and non-structural carbohydrates (NSC). Temperature and N both affected algal growth, and there were temperature × N interactions, which were sometimes affected by presence/absence of zooplankton. For example, cyanobacteria (but not green algae) growth increased with both temperature and N, especially from 25°C to 30°C, but N and herbivore presence increased cyanobacterial growth primarily only at 30°C. In general, temperature and N had little consistent effect on NSC, but increasing temperature and N tended to increase protein content in Microcystis and Anabaena (temperature effects mostly at higher N levels). In Anabaena, increases in N did not increase growth or protein at 20°C or 25°C, but did increase both at 30°C, indicating that N fixation is damaged at high temperatures and that high NO3 can overcome this damage. These results indicate that future global warming and continued eutrophication will increase cyanobacterial growth, as well influence algal herbivory and competition between N-fixing and non-N-fixing cyanobacteria.

Nitrogen Constrains the Growth of Late Summer Cyanobacterial Blooms in Lake Erie

Phosphorus (P) is generally considered to be the main limiting nutrient to freshwater phytoplankton productivity. However, recent research is drawing attention to the importance of nitrogen (N) in freshwater eutrophication and N often constrains growth of cyanobacteria in small lakes. In this study we determined phytoplankton nutrient limitation in a large lake, Lake Erie during two growing seasons. During 2010 and 2011, nutrient enrichment bioassays (+P, +N and, +P and N) were conducted monthly from June through September with water collected in Maumee Bay (site MB18) and in the center of the western basin (site WBC). Nutrient concentrations were monitored every other week. At MB18, total P concentration was often >3 mmol/L and nitrate concentration decreased from >250 mmol/L in early summer to mmol/L in late summer. Nitrogen and P levels were about five-fold less at WBC. Bioassays indicated that phytoplankton nutrient limitation varied in summer, spatially, and even among phytoplankton groups. For site MB18, +P increased chlorophyll concentration in one of the eight bioassays, indicating that P did not typically limit production. For site WBC, +P increased chlorophyll concentration in six of the eight bioassays. As a result of very low ambient nitrate concentration (mmol/L) in late summer, +N (without P) increased chlorophyll concentration, suggesting symptoms of N-limitation. The N-fixing cyanobacterium Anabaena became dominant following N-limitation. This study highlights the need to reduce P loading to restore water quality. Furthermore, due to low nitrate concentration, the severity of the cyanobacterial blooms could be worse if not for N-limitation in western Lake Erie.

Induced Accumulation of Polyphenolics and Flavonoids in Cyanobacteria under Salt Stress Protects Organisms through Enhanced Antioxidant Activity

Induced accumulation of polyphenolics and flavonoids in cyanobacterial strains grown under different salt concentrations is correlated with their growth under stress conditions and enhanced antioxidant activity. Plectonema boryanum, Hapalosiphon intricatus, Anabaena doliolum and Oscillatoria acuta grown for 21 days under different salt concentrations (80, 160, 240, 320 and 400 mM) in BG11 medium showed differential growth responses in terms of biomass, total protein, chlorophyll content, total content of polyphenol, flavonoid and carotenoid, accumulation of phenolic acids (gallic, caffeic, chlorogenic, ferullic and vanillic) and flavonoids (rutin and quercetin). Cyanobacterial extracts showed prominent free radical scavenging antioxidant activity (AOA) in terms of % DPPH discoloration. Highly significant (p < 0.05) and strong correlation was found between TPC and AOA (r = 0.974). Other positive but non-significant (p < 0.05) correlations were observed between AOA and gallic acid (r = 0.893) and AOA and caffeic acid (r = 0.931). Significant and strong correlation was also observed between gallic and caffeic acid (r = 0.973). Positive but lesser magnitude correlations were recorded between TPC and caffeic acid (r = 0.905), TPC and gallic acid (r = 0.920), gallic and vanillic acid (r = 0.916) and caffeic and vanillic acid (r = 0.814). An integrated combination of growth parameters, salt-induced accumulation of phenylpropanoids and stress-derived subsequent antioxidant property of cyanobacterial extracts is thought to provide evidence that secondary metabolic changes can act as the possible alternative mechanism to overcome stress-induced damages in cells.