The first study on the effects of microcystin-RR on gene expression profiles of antioxidant enzymes and heat shock protein-70 in Synechocystis sp PCC6803

Microcystins are heptapeptide toxins produced by cyanobacteria. Microcystin-RR(MC-RR) is a common variant among the 80 variants identified so far. There have been many investigations documenting the toxic effects of microcystins on animals and higher plants, but little is known on the toxic effects of microcystins on algae, especially at molecular level. We studied the effects of MC-RR on gene expression profile of a few antioxidant enzymes and heat shock protein-70 (Hsp70) in Synechocystis sp. PCC6803. After two days post-exposure, a high dose toxin (5 mg/l, about 4.8 x 10(-3) mM) significantly increased expression levels of the genes gpx1, sodB, katG, acnB, gamma-TMTand dnaK2, while a relatively low dose toxin (1 mg/l, about 9.63 x 10(-4) mM) induced a moderate and slow increase of gene expression. Our results indicate that MC-RR could induce the oxidative stress in Synechocystis sp. PCC6803 and the increase in gene expression of antioxidant enzymes and Hsp70 might protect the organism from the oxidative damage. in addition, cell aggregation was observed during the early period of exposure, which might be a specific oxidative stress reaction to MC-RR. (C) 2008 Elsevier Ltd. All rights reserved.

A high throughput Nile red method for quantitative measurement of neutral lipids in microalgae

Isolation of high neutral lipid-containing microalgae is key to the commercial success of microalgae-based biofuel production. The Nile red fluorescence method has been successfully applied to the determination of lipids in certain microalgae, but has been unsuccessful in many others, particularly those with thick, rigid cell walls that prevent the penetration of the fluorescence dye. The conventional "one sample at a time" method was also time-consuming. In this study, the solvent dimethyl sulfoxide (DMSO) was introduced to microalgal samples as the stain carrier at an elevated temperature. The cellular neutral lipids were determined and quantified using a 96-well plate on a fluorescence spectrophotometer with an excitation wavelength of 530 nm and an emission wavelength of 575 run. An optimized procedure yielded a high correlation coefficient (R-2 = 0.998) with the lipid standard triolein and repeated measurements of replicates. Application of the improved method to several green algal strains gave very reproducible results with relative standard errors of 8.5%, 3.9% and 8.6%, 4.5% for repeatability and reproducibility at two concentration levels (2.0 mu g/mL and 20 mu g/mL), respectively. Moreover, the detection and quantification limits of the improved Nile red staining method were 0.8 mu g/mL and 2.0 mu g/mL for the neutral lipid standard triolein, respectively. The modified method and a conventional gravimetric determination method provided similar results on replicate samples. The 96-well plate-based Nile red method can be used as a high throughput technique for rapid screening of a broader spectrum of naturally-occurring and genetically-modified algal strains and mutants for high neutral lipid/oil production. (C) 2009 Published by Elsevier B.V.

Grazing on toxic and non-toxic Microcystis aeruginosa PCC7820 by Unio douglasiae and Corbicula fluminea

To explore the potential grazing effects of mussels on Microcystis aeruginosa, a common bloom-forming phytoplankton, Unio douglasiae and Corbicula fluminea were fed with Scenedesmus obliquus, toxic and non-toxic strains of Microcystis aeruginosa as single food and as mixtures in the laboratory. When fed with single foods, U. douglasiae has similar clearance rates on the three algae populations, while C. fluminea has significantly lower clearance rate on toxic M. aeruginosa than those on the other two algae populations. When fed with mixture foods, both the mussels show significantly higher clearance rates than on single foods. The clearance rates of U. douglasiae on the different food mixtures are not significantly different, and C. fluminea has a significantly lower clearance rate on the toxic food mixtures than that on non-toxic food mixtures. Although the relative lower clearance rates of C. fluminea on toxic food, we may still deduce that both the mussels can exert grazing pressure on phytoplankton. The deduction is supported by the composition of the excretion products. The excretion products (faeces and pseudofaeces) of both mussels contained mainly S. obliquus. In both mixed-food treatments, the ratios of S. obliquus to M. aeruginosa in the excrete products are significantly higher than those in the foods. Therefore, it can be concluded that both mussels prefer M. aeruginosa to S. obliquus, and can cause grazing pressure on M. aeruginosa.

Effects of arsenate on the growth and microcystin production of Microcystis aeruginosa isolated from Taiwan as influenced by extracellular phosphate

Arsenic pollution and eutrophication are both prominent issues in the aquaculture ponds of Taiwan. It is important to study the effects of arsenic on algal growth and toxin production in order to assess the ecological risk of arsenic pollution, or at least to understand naturally occurring ponds. The sensitivity of algae to arsenate has often been linked to the structural similarities between arsenate and phosphate. Thus, in this study we examined the effects of arsenate (10(-8) to 10(-4) M) on Microcystis aeruginosa TY-1 isolated from Taiwan, under two phosphate regimes. The present study showed that M. aeruginosa TY-1 was arsenate tolerant up to 10(-4) M, and that this tolerance was not affected by extracellular phosphate. However, it seems that extracellular phosphate contributed to microcystin production and leakage by M. aeruginosa in response to arsenate. Under normal phosphate conditions, total toxin yields after arsenate treatment followed a typical inverted U-shape hormesis, with a peak value of 2.25 +/- 0.06 mg L-1 in the presence of 10(-7) M arsenate, whereas 10(-8) to 10(-6) M arsenate increased leakage of similar to 75% microcystin. Under phosphate starvation, total toxin yields were not affected by arsenate, while 10(-6) and 10(-5) M arsenate stimulated microcystin leakage. It is suggested that arsenate may play a role in the process of microcystin biosynthesis and excretion. Given the arsenic concentrations in aquaculture ponds in Taiwan, arsenate favors survival of toxic M. aeruginosa in such ponds, and arsenate-stimulated microcystin production and leakage may have an impact on the food chain.

Changes in benthic algal communities following construction of a run-of-river dam

Ecological responses to dam construction are poorly understood, especially for downstream benthic algal communities. We examined the responses of benthic algal communities in downstream reaches of a tributary of the Xiangxi River, China, to the construction of a small run-of-river dam. From February 2003 to August 2006, benthic algae, chemical factors, and habitat characteristics were monitored upstream and downstream of the dam site. This period spanned 6 mo before dam construction and 37 mo after dam construction. Benthic algal sampling yielded 199 taxa in 59 genera that belonged to Bacillariophyta, Chlorophyta, and Cyanophyta. Some physical factors (flow velocity, water depth, and channel width) and 3 algal metrics (diatom species richness, Margalef diversity, and % erect individuals) were significantly affected by the dam construction, whereas chemical factors (e.g., NH4-N, total N, SiO2) were not. Nonmetric multidimensional scaling (NMS) ordinations showed that overall algal assemblage structure downstream of the dam sites was similar to that of upstream control sites before dam construction and for 1 year after dam construction (p > 0.05). However, sites belonging to upstream and downstream reaches were well separated on NMS axis 1 during the 2(nd) and 3(rd) years after dam construction. Our results suggest that impacts of dam construction on benthic algal communities took 2 to 3 y to emerge. Further development of a complete set of indicators is needed to address the impact of small-dam construction. Our observations underscore the need for additional studies that quantify ecological responses to dam construction over longer time spans.

UV-B-induced Oxidative Damage and Protective Role of Exopolysaccharides in Desert Cyanobacterium Microcoleus vaginatus

UV-B-induced oxidative damage and the protective effect of exopolysaccharides (EPS) in Microcoleus vaginatus, a cyanobacterium isolated from desert crust, were investigated. After being irradiated with UV-B radiation, photosynthetic activity (Fv/Fm), cellular total carbohydrates, EPS and sucrose production of irradiated cells decreased, while reducing sugars, reactive oxygen species (ROS) generation, malondialdehyde (MDA) production and DNA strand breaks increased significantly. However, when pretreated with 100 mg/L exogenous EPS, EPS production in the culture medium of UV-B stressed cells decreased significantly; Fv/Fm, cellular total carbohydrates, reducing sugars and sucrose synthase (SS) activity of irradiated cells increased significantly, while ROS generation, MDA production and DNA strand breaks of irradiated cells decreased significantly. The results suggested that EPS exhibited a significant protective effect on DNA strand breaks and lipid peroxidation by effectively eliminating ROS induced by UV-B radiation in M. vaginatus.

Microcystin-RR induces physiological stress and cell death in the cyanobacterium Aphanizomenon sp DC01 isolated from Lake Dianchi, China

The phytoplankton community in Lake Dianchi (Yunnan Province, Southwestern China) is dominated in April by a bloom of Aphanizomenon, that disappears Suddenly and is displaced by a Microcystis bloom in May. The reasons for the rapid bloom disappearance phenomenon and the temporal variability in the composition of phytoplankton assemblages are poorly understood. Cell growth, ultrastructure and physiological changes were examined in cultures of Aphanizomenon sp. DC01 isolated from Lake Dianchi exposed to different closes of rnicrocystin-RR (MC-RR) produced by the Microcystis bloom. MC-RR concentrations above 100 mu g L-1 markedly inhibited the pigment (chlorophyll-a, phycocyanin) synthesis and caused an increase of soluble carbohydrate and protein contents and nitrate reductase activity of toxin-treated blue-green algae. A drastic. reduction in photochemical efficiency of PSII (Fv/Fm) was also found. Morphological examinationn showed that the Aphanizomenon filaments disintegrated and file cells lysed gradually after 48 h Of toxin exposure. Transmission electron microscopy revealed that cellular inclusions of stressed cells almost leaked out completely and the cell membranes were grossly damaged. These findings demonstrate the allelopathic activity of Microcystis aeruginosa inducing physiological stress and cell death of Aphanizomenon sp. DC01 Although the active concentrations of microcystin were rather high, we propose that microcystin may function as allelopathic Substance due to inhomogeneous toxin concentrations close to Microcystis cells. Hence, it may play a role in species Succession of Aphanizomenon and Microcystis in Lake Dianchi.

A Review of Nontraditional Biomanipulation

The aim of this review is to identify problems, find general patterns, and extract recommendations for successful management using nontraditional biomanipulation to improve water quality. There are many obstacles that prevent traditional biomanipulation from achieving expectations: expending largely to remove planktivorous fish, reduction of external and internal phosphorus, and macrophyte re-establishment. Grazing pressure from large zooplankton is decoupled in hypereutrophic waters where cyanobacterial blooms flourish. The original idea of biomanipulation (increased zooplankton grazing rate as a tool for controlling nuisance algae) is not the only means of controlling nuisance algae via biotic manipulations. Stocking phytoplanktivorous fish may be considered to be a nontraditional method; however, it can be an effective management tool to control nuisance algal blooms in tropical lakes that are highly productive and unmanageable to reduce nutrient concentrations to low levels. Although small enclosures increase spatial overlap between predators and prey, leading to overestimates of the impact of predation, microcosm and whole-lake experiments have revealed similar community responses to major factors that regulate lake communities, such as nutrients and planktivorous fish. Both enclosure experiments and large-scale observations revealed that the initial phytoplankton community composition greatly impacted the success of biomanipulation. Long-term observations in Lake Donghu and Lake Qiandaohu have documented that silver carp (Hypophthalmichthys molitrix) and bighead carp (H. nobilis) (two filter-feeding planktivorous species commonly used in management) can suppress Microcystis blooms efficiently. The introduction of silver and bighead carp could be an effective management technique in eutrophic systems that lack macrozooplankton. We confirmed that nontraditional biomanipulation is only appropriate if the primary aim is to reduce nuisance blooms of large algal species, which cannot be controlled effectively by large herbivorous zooplankton. Alternatively, this type of biomanipulation did not work efficiently in less eutrophic systems where nanophytoplankton dominated.

Enhanced protection against oxidative stress in an astaxanthin-overproduction Haematococcus mutant (Chlorophyceae)

Many unicellular green algae can become yellow or red in various natural habitats due to mass accumulation of a secondary carotenoid, such as lutein, or astaxanthin. The accumulation of secondary carotenoids is generally thought to be a survival strategy of the algae under photo-oxidative stress or other adverse environmental conditions. The physiological role of the carotenoids in stress response is less well understood at the subcellular or molecular level. In this study, a stable astaxanthin overproduction mutant (MT 2877) was isolated by chemical mutagenesis of a wild type (WT) of the green microalga Haematococcus pluvialis Flotow NIES-144. MT 2877 was identical to the WT with respect to morphology, pigment composition, and growth kinetics during the early vegetative stage of the life cycle. However, it had the ability to synthesize and accumulate about twice the astaxanthin content of the WT under high light, or under high light in the presence of excess amounts of ferrous sulphate and sodium acetate. Under stress, the mutant exhibited higher photosynthetic activities than the WT, based on considerably higher chlorophyll fluorescence induction, chlorophyll autofluorescence intensities, and oxygen evolution rates. Cell mortality caused by stress was reduced by half in the mutant culture compared with the WT. Enhanced protection of the mutant against stress is attributed to its accelerated carotenogenesis and accumulation of astaxanthin. Our results suggest that MT 2877, or other astaxanthin overproduction Haematococcus mutants, may offer dual benefits, as compared with the wild type, by increasing cellular astaxanthin content while reducing cell mortality during stress-induced carotenogenesis.

Effects of gibberellin A(3) on growth and microcystin production in Microcystis aeruginosa (cyanophyta)

Environmental. factors that affect the growth and microcystin production of microcystis have received worldwide attention because of the hazards microcystin poses to environmental safety and public health. Nevertheless, the effects of organic anthropogenic pollution on microcystis are rarely discussed. Gibberellin A(3) (GA(3)) is a vegetable hormone widely used in agriculture and horticulture that can contaminate water as an anthropogenic pollutant. Because of its common occurrence, we studied the effects of GA3 on growth and microcystin production of Microcystis aeruginosa (M. aeruginosa) PCC7806 with different concentrations (0.001-25mg/L) in batch culture. The control was obtained without gibberellin under the same culture conditions. Growth, estimated by dry weight and cell number, increased after the GA3 treatment. GA3 increased the amounts of chlorophyll a, phycocyanin and cellular-soluble protein in the cells of M. aeruginosa PCC7806, but decreased the accumulation of water-soluble carbohydrates. In addition, GA3 was observed to affect nitrogen absorption of the test algae, but to have no effect on the absorption of phosphorus. The amount of microcystin measured by enzyme-Linked immunosorbent assay (ELISA) increased in GA3 treatment groups, but the stimulatory effects were different in different culture phases. It is suggested that GA3 increases M. aeruginosa growth by stimulating its absorbance of nitrogen and increasing its ability to use carbohydrates, accordingly increasing cellular pigments and thus finally inducing accumulation of protein and microcystin. (C) 2007 Elsevier GmbH. All rights reserved.

Nitrogen dynamics in large shallow eutrophic Lake Chaohu, China

Temporal and spatial dynamics of nitrogen in lake and interstitial water were studied monthly in a large shallow, eutrophic lake in subtropical China from October 2002 to September 2003. The distribution of nitrogen was consistent with the idea that high nitrogen concentrations in the western part of the lake resulted from high levels of the nutrients from the surrounding cities through sewage-drainage systems. Nitrate was the predominant form of nitrogen in the overlying water, while ammonium was predominant in the interstitial water, indicating that strong oxidative nutrient regeneration occurred near the sediment-water interface. Nitrate could be an important dissolved inorganic matter source for phytoplankton, which in turn influenced the seasonal variations of nitrate concentrations in lake water. Significant positive correlation between ammonium fluxes and water temperature was observed and could probably be attributed to the intensified ammonification and nitrate reduction with increased temperature. Positive correlation between ammonium fluxes and algae biomass and Chl a concentrations may indicate that phytoplankton was an important factor driving ammonium fluxes in our study lake, and vice versa that higher fluxes of ammonium supported a higher biomass of the phytoplankton.

Control of lunar and martian dust - Experimental insights from artificial and natural cyanobacterial and algal crusts in the desert of Inner Mongolia, China

Studies on the colonization of environmentally extreme ground surfaces were conducted in a Mars-like desert area of Inner Mongolia, People's Republic of China, with microalgae and cyanobacteria. We collected and mass-cultured cyanobacterial strains from these regions and investigated their ability to form desert crusts artificially. These crusts had the capacity to resist sand wind erosion after just 15 days of growth. Similar to the surface of some Chinese deserts, the surface of Mars is characterized by a layer of fine dust, which will challenge future human exploration activities, particularly in confined spaces that will include greenhouses and habitats. We discuss the use of such crusts for the local control of desert sands in enclosed spaces on Mars. These experiments suggest innovative new directions in the applied use of microbe-mineral interactions to advance the human exploration and settlement of space.

A simple closed aquatic ecosystem (CAES) for space

A closed aquatic ecosystem (CAES) was developed to stud), the effects of microgravity on the function of closed ecosystems aboard the Chinese retrieved satellite and on the spacecraft SHENZHOU-II. These systems housed a small freshwater snail (Bulinus australianus) and an autotrophic green algae (Chlorella pyrenoidosa). The results of the test on the satellite were that the concentration of algae changed little, but that the snails died during the experiments. We then sought to optimize the function of the control system, the cultural conditions and the data acquisition system and carried out an experiment on the spacecraft SHENZHOU-II. Using various sensors to monitor the CAES, real-time data regarding the operation of the CAES in microgravity was acquired. In addition, all on-board Ig centrifuge was included to identify gravity-related factors. It was found that microgravity is the major factor affecting the operation of the CAES in space. The change in biomass of the primary producer during each day in microgravity was larger than that of the control groups. The mean biomass concentration per day in the microgravity group decreased, but that of the control groups increased for several days and then leveled off. Space effects on the biomass of a primary producer may be a result of microgravity effects leading to increasing metabolic rates of the consumer combined with decreases in photosynthesis. (c) 2007 COSPAR. Published by Elsevier Ltd. All rights reserved.

Reduction in microcystin concentrations in large and shallow lakes: Water and sediment-interface contributions

Blooms of cyanobacteria, or blue-greens, are known to produce chemicals, such as microcystins, which can be toxic to aquatic and terrestrial organisms. Although previous studies have examined the fate of microcystins in freshwater lakes, primary elimination pathways and factors affecting degradation and loss have not been fully explained. The goal of the present study was to explore sources of algal toxins and investigate the distribution and biodegradation of microcystins in water and sediment through laboratory and field analyses. Water and sediment samples were collected monthly from several locations in Lake Taihu from February 2005 to January 2006. Samples were analyzed for the presence of microcystin. Water and sediment were also used in laboratory studies to determine microcystin degradation rates by spiking environmental samples with known concentrations of the chemical and observing concentration changes over time. Some water samples were found to efficiently degrade microcystins. Microcystin concentrations dropped faster in water collected immediately above lake sediment (overlying water). Degradation in sediments was higher than in water. Based on spatial distribution analyses of microcystin in Lake Taihu, higher concentrations (relative to water concentrations) of the chemical were found in lake sediments. These data suggest that sediments play a critical role in microcystin degradation in aquatic systems. The relatively low levels of microcystins found in the environment are most likely due to bacterial biodegradation. Sediments play a crucial role as a source (to the water column) of bio-degrading bacteria and as a carbon-rich environment for bacteria to proliferate and metabolize microcystin and other biogenic toxins produced by cyanobacteria. These, and other, data provide important information that may be applied to management strategies for improvement of water quality in lakes, reservoirs and other water bodies. (C) 2007 Elsevier Ltd. All rights reserved.

The response of antioxidant systems in Nostoc sphaeroides against UV-B radiation and the protective effects of exogenous antioxidants

UV radiation is one of many harmful factors found in space that are detrimental to organisms on earth in space exploration. In the present work, we examined the role of antioxidant system in Nostoc sphaeroides Kutz (Cyanobacterium) and the effects of exogenously applied antioxidant molecules on its photosynthetic rate under UV-B radiation. It was found that UV-B radiation promoted the activity of antioxidant system to protect photosystem 11 (PSII) and exogenously applied antioxidant: sodium nitroprusside (SNP) and N-acetylcysteine (NAC) had an obvious protection on PSII activity under UV-B radiation. The activity of superoxide dismutase (SOD, EC 1.15.1.1), catalase (CAT, EC 1.11.1.6), peroxidase (POD, EC 1.11.1.7) and content of NIDA (malondialdehyde) and ASC (ascorbate) were improved by 0.5 mM and 1 mM SNP, but 0.1 mM SNP decreased the activity of antioxidant system. Addition of exogenous NAC decreased the activity of SOD, POD, CAT and the content MDA and ASC. In contrast, exogenously applied NAC increased GSH content. The results suggest that exogenous SNP and NAC may protect algae by different mechanisms: SNP may play double roles as both sources of reactive free radicals as well as ROS scavengers in mediating the protective role of PSII on algae under UV-B radiation. On the other hand, NAC functions as an antioxidant or precursor of glutathione, which could protect PSII directly from UV-B radiation. (c) 2007 COSPAR, Published by Elsevier Ltd. All rights reserved.