Allelopathic mechanism of pyrogallol to Microcystis aeruginosa PCC7806 (Cyanobacteria): From views of gene expression and antioxidant system

Pyrogallol is a potent allelochemical on Microcystis aeruginosa, but its allelopathic mechanism is not fully known. In order to explore this mechanism, gene expressions for prx, mcyB, psbA, recA, grpE, fabZ under pyrogallol stress were studied, and activities of the main antioxidant enzymes were also measured. The results showed that expression of grpE and recA showed no significant change under pyrogallol stress, while psbA and mcyB were up-regulated at 4 mg L-1. Both prx and fabZ were up-regulated even under exposure to 1 mg L-1 pyrogallol concentration. The activities of superoxide dismutase (SOD) and catalase (CAT) were enhanced under pyrogallol stress. Levels of malodialdehyde (MDA) at 2 and 4 mg L-1 pyrogallol were significantly higher than those of the controls. It was concluded that oxidant damage is an important mechanism for the allelopathic effect of pyrogallol on M. aeruginosa. (c) 2009 Elsevier Ltd. All rights reserved.

Plasma biochemical responses of the planktivorous filter-feeding silver carp (Hypophthalmichthys molitrix) and bighead carp (Aristichthys nobilis) to prolonged toxic cyanobacterial blooms in natural waters

The planktivorous filter-feeding silver carp (Hypophthalmichthys molitrix) and bighead carp (Aristichthys nobilis) are the attractive candidates for bio-control of plankton communities to eliminate odorous populations of cyanobacteria. However, few studies focused on the health of such fishes in natural water body with vigorous toxic blooms. Blood parameters are useful and sensitive for diagnosis of diseases and monitoring of the physiological status of fish exposed to toxicants. To evaluate the impact of toxic cyanobacterial blooms on the planktivorous fish, 12 serum chemistry variables were investigated in silver carp and bighead carp for 9 months, in a large net cage in Meiliang Bay, a hypereutrophic region of Lake Taihu. The results confirmed adverse effects of cyanobacterial blooms on two phytoplanktivorous fish, which mainly characterized with potential toxicogenomic effects and metabolism disorders in liver, and kidney dysfunction. In addition, cholestasis was intensively implied by distinct elevation of all four related biomarkers (ALP, GGT, DBIL, TBIL) in bighead carp. The combination of LDH, AST activities and DBIL, URIC contents for silver carp, and the combination of ALT. ALP activities and TBIL, DBIL. URIC concentrations for bighead carps were found to most strongly indicate toxic effects from cyanobacterial blooms in such fishes by a multivariate discriminant analysis. (C) 2009 Elsevier B.V. All rights reserved.

EFFECTS OF IRON ON PHYSIOLOGICAL AND BIOCHEMICAL CHARACTERISTICS OF Microcystis wesenbergii (Kom.) Kom. (Cyanobacterium)

Iron is an essential trace element for biological requirements of phytoplankton. Effects of iron on physiological and biochemical characteristics of Microcystis wesenbergii were conducted in this study. Results showed that 0.01 mu M [Fe3+] seriously inhibited growth and chlorophyll synthesis of M. wesenbergii, and induced temporary increase of ATPase activities, however, NR. ACP and ALP activities were restrained by iron limitation. Interestingly, iron addition on day 8 resulted in the gradual restoration of structures and functions of above enzymes and resisted a variety of stresses from iron limitation. M. wesenbergii in 10 mu M [Fe3+] treatment group grew normally. enzymes maintained normal levels, and residual phosphate contents in cultures first sharply decreased, then smoothly as M. wesenbergii has a characteristic of luxury consumption of phosphorus. Above parameters in 100 mu M [Fe3+] treatment group were almost same with those in 10 mu M [Fe3+] treatment group except for NR, ACP and ALP activities. In 100 mu M [Fe3+] treatment group, activities of ACP and ALP had temporary increase because phosphate and ferric iron could form insoluble compound - ferric phosphate (Fe3PO4) through adsorption effect. resulting in lack of bioavailable phosphate in culture media. The experiment suggested that too low or too high iron can affect obviously physiological and biochemical characteristics of M. wesenbergii.

Food Consumption by In Situ Pen-Cultured Planktivorous Fishes and Effects on an Algal Bloom in Lake Taihu, China

Silver carp (Hypophthalmichthys molitrix) and bighead carp (Aristichthys nobilis) were used as a new pen-cultureed biomanipulation technique to control algal blooms in Meiliang Bay of Lake Taihu. In order to evaluate the capacity of these two fishes to decrease algal blooms, diel feeding samplings were carried out in May (without algal blooms) and September (with algal blooms) in 2005. Based on estimated food consumption by the Elliott-Persson model, silver carp increased daily food consumption from 2.07 g dry weight per 100 g wet body weight in May before the outbreak of algal blooms to 4.98 g dry weight per 100 g wet body weight in September during algal blooms outbreak. However, no obvious variation of food consumption was observed in bighead carp during the study period. This species 1.88 and 1.54 g dry weight of plankton per 100 g wet body weight in May and September, respectively. Silver carp had a higher feeding capacity for plankton than bighead carp. Biotic factors (i.e., fish size and conspecific competition with natural species in the lake) may affect the feeding behaviors of both carps as well as seasonal variation of plankton communities in the pen.

Effects of Arsenate on Microcystin Content and Leakage of Microcystis Strain PCC7806 Under Various Phosphate Regimes

Both arsenic pollution and eutrophication are prominent environmental issues when considering the problem of global water pollution. It is important to reveal the effects of arsenic species on cyanobacterial growth and toxin yields to assess ecological risk of arsenic pollution or at least understand naturally occurring blooms. The sensitivity of cyanobacteria to arsenate has often been linked to the structural similarities of arsenate and phosphate. Thus, we approached the effect of arsenate with concentrations from 10(-8) to 10(-4) M on Microcystis strain PCC7806 under various phosphate regimes. The present study showed that Microcystis strain PCC7806 was arsenate tolerant up to 10(-4) M. And such tolerance was without reference to both content of intra- and extra-cellular phosphate. It seems that arsenate involved the regulation of microcystin synthesis and cellular polyphosphate contributed to microcystin production of Microcystis responding to arsenate, since there was a positive linear correlation of the cellular microcystin quota with the exposure concentration of arsenate when the cells were not preconditioned to phosphate starvation. It is presumed that arsenate could help to actively export microcystins from living Microcystis cells when preconditioned to phosphate starvation and incubated with the medium containing 1 mu M phosphate. This study firstly provided evidence that microcystin content and/or release of Microcystis might be impacted by arsenate if it exists in harmful algal blooms. (C) 2008 Wiley Periodicals, Inc. Environ Toxicol 24:97 94, 2009.

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.

Non-microcystin producing Microcystis wesenbergii (Komarek) Komarek (Cyanobacteria) representing a main waterbloom-forming species in Chinese waters

It is well known that several morphospecies of Microcystis, such as Microcystis aeruginosa (Kutzing) Lemmermann and Microcystis viridis (A. Brown) Lemmermann can produce hepatotoxic microcystins. However, previous studies gave contradictory conclusions about microcystin production of Microcystis wesenbergii (Komarek) Komarek. In the present study, ten Microcystis morphospecies were identified in waterblooms of seven Chinese waterbodies, and Microcystis wesenbergii was shown as the dominant species in these waters. More than 250 single colonies of M. wesenbergii were chosen, under morphological identification, to examine whether M. wesenbergii produce hepatotoxic microcystin by using multiplex PCR for molecular detection of a region (mcyA) of microcystin synthesis genes, and chemical analyses of microcystin content by ELISA and HPLC for 21 isolated strains of M. wesenbergii from these waters were also performed. Both molecular and chemical methods demonstrated that M. wesenbergii from Chinese waters did not produce microcystin. (C) 2007 Elsevier Ltd. All rights reserved.

In vivo studies on the toxic effects of microcystins on mitochondrial electron transport chain and ion regulation in liver and heart of rabbit

This study examined the toxic effects of microcystins on mitochondria of liver and heart of rabbit in vivo. Rabbits were injected i.p. with extracted microcystins (mainly MC-RR and -LR) at two doses, 12.5 and 50 MCLReq. mu g/kg bw, and the changes in mitochondria of liver and heart were studied at 1, 3,12, 24 and 48 h after injection. MCs induced damage of mitochondrial morphology and lipid peroxidation in both liver and heart. MCs influenced respiratory activity through inhibiting NADH dehydrogenase and enhancing succinate dehydrogenase (SDH). MCs altered Na+-K+-ATPase and Ca2+-Mg2+-ATPase activities of mitochondria and consequently disrupted ionic homeostasis, which might be partly responsible for the loss of mitochondrial membrane potential (MMP). MCs were highly toxic to mitochondria with more serious damage in liver than in heart. Damage of mitochondria showed reduction at 48 h in the low dose group, suggesting that the low dose of MCs might have stimulated a compensatory response in the rabbits. (C) 2008 Elsevier Inc. All rights reserved.

Studies on the toxic effects of La3+ to Tetrahymena thermophila by microcalorimetry

The toxic effects of La3+ on Tetrahymena thermophila have been studied by microcalorimetry at 28 degrees C. The metabolic rate constant (r) and peak time were linked to the concentration of La3+. The changes of metabolic rate constant indicated that low-concentration La3+ (0-75 mg/L) had no significant effects on the metabolism of Tetrahymena cells but high-concentration La3+ (100-175 mg/L) could inhibit their metabolism. From the results obtained by cell counting and fluorescence depolarization measurements, the inhibition of metabolism resulted from the decrease in cell number and the reduction in cell membrane fluidity. According to the results, it is clear that the metabolic mechanism of Tetrahymena cells has been changed with the addition of high-concentration La3+. In addition, microcalorimetry of Tetrahymena could be a sensible, easy-to-use, and convenient method for monitoring the potential effects of rare earth elements on cells and the freshwater ecosystem.

Polyphasic characterization of water bloom forming Raphidiopsis species (cyanobacteria) from central China

Two strains of Raphidiopsis Fritsch et Rich were isolated from a fishpond in Wuhan city, China and rendered axenic, and characterized by a combination of morphological, physiological, biochemical and genetic methods. Morphologically the strains were identified as Raphidiopsis mediterranea Skuja (straight trichomes) and R. curvata Fritsch et Rich (coiled trichomes). These two strains demonstrated slight differences in optimal temperature range and GC content, while sharing some common characteristics including inability to grow hetertrophically, similar salinity tolerance (up to 0.78%) and an identical fatty acid composition. Cyanotoxins were not found in the strain of R. mediterranea, however, the strain of R. curvata contained both deoxycylindrospermopsin and cylindrospermopsin. Phylogenetic affiliations inferred from 16S rRNA gene sequences demonstrated that both Raphidiopsis strains clustered with Cylindrospermopsis, demonstrating their phylogenetic ties to Nostocaceae. (c) 2007 Elsevier B.V. All rights reserved.

Stable carbon isotope variations in surface bloom scum and subsurface seston among shallow eutrophic lakes

Carbon stable isotope analysis of surface bloom scum and subsurface seston samples was conducted in shallow eutrophic lakes in China during warm seasons from 2003 to 2004. delta C-13 values of bloom scum were always higher (averaged 5 parts per thousand) than those of seston in this study, and the possible reasons were attributed to (i) direct use of atmospheric CO2 at the air-water interface, (ii) decrease in C-13 fractionation due to higher carbon fixation, (iii) active CO2 transport, and/or (iv) HCO3 accumulation. Negative correlation between delta C-13(scum) - delta C-13(seston) and pH in the test lakes indicated that phytoplankton at the subsurface water column increased isotopic enrichment under the-carbon limitation along with the increase of pH, which might in turn decreased the differences in 313 C between the subsurface seston and the surface scums. Significant positive correlations of seston 8 13C with total concentrations of nitrogen and phosphorus in water column suggested that the increase in delta C-13 of seston with trophic state was depending on nutrient (N or P, or both) supply. Our study showed that delta C-13 of phytoplankton was indicative of carbon utilization, primary productivity, and nutrient supply among the eutrophic lakes. (C) 2007 Elsevier B.V All rights reserved.

Microcalorimetric study on the toxic effect of Pb2+ to Tetrahymena

The toxic effect of Pb2+ has been studied in eukaryotic cells by using Tetrahymena as a target. The maximum power (P (m)) and the growth rate constant (k) were determined, which showed that values of P (m) and k were linked to the concentration (C) of Pb2+. The addition of Pb2+ caused a decrease of the maximum heat production and growth rate constant, indicating that Tetrahymena growth was inhibited in the presence of Pb2+, and Pb2+ took part in the metabolism of cells. From micrographs, morphological changes of Tetrahymena were observed with addition of Pb2+, indicating that the toxic effect of Pb2+ derived from destroying the membrane of surface of Tetrahymena. According to the thermogenic curves and photos of Tetrahymena under different conditions, it is clear that metabolic mechanism of Halobacterium halobium R1 growth has been changed with the addition of Pb2+.

Toxicity of microcystins in the isolated hepatocytes of common carp (Cyprinus carpio L.)

The toxicity of hepatotoxic microcystins produced mainly by Microcystis aeruginosa in mammals and fishes was well studied in recent years. However, there were scarcely reports in toxic effects of microcystins on isolated hepatocytes of fishes, especially investigation of microcystin-induced apoptosis and/or necrosis in carp hepatocytes. In the present study, the isolated hepatocytes of common carp were exposed to various concentrations of microcystins (0.01, 0.1, 1, 10, 100, 1000 mu g L-1) for 2, 4, 8, 16 and 24 h, respectively, and cytotoxicity of microcystins in the toxin-treated cells was determined. Results of this study showed that cytotoxicity of microcystins on carp hepatocytes was time and dose-dependent, and the approximate LC50 of microcystins in carp hepatocytes was 169.2 mu g L-1. The morphological changes typical of apoptosis, such as blebbing of cell membrane, condensation and fragmentation of cell nucleus were observed in the hepatocytes exposed to microcystins (1, 10 and 100 mu g L-1) using fluorescence and differential interference contrast microscopy. Agarose gel electrophoresis of DNA demonstrated a typical apoptotic "ladder pattern" in microcystin-treated hepatocytes after 16 h of exposure. Results of the present study indicated that the form of cell death in microcystin-treated hepatocytes depend on the exposure dose of toxin. When lower concentration of microcystins (10 and 100 mu g L-1) was used for exposure, carp hepatocytes died in apoptosis while, when higher one used (1000 mu g L-1), they died in the form of necrosis. (C) 2006 Elsevier Inc. All rights reserved.

Effects of iron on growth, pigment content, photosystem II efficiency, and siderophores production of Microcystis aeruginosa and Microcystis wesenbergii

Changes in growth, photosynthetic pigments, and photosystem II (PS II) photochemical efficiency as well as production of siderophores of Microcystis aeruginosa and Microcystis wesenbergii were determined in this experiment. Results showed growths of M. aeruginosa and M. wesenbergii, measured by means of optical density at 665 nm, were severely inhibited under an iron-limited condition, whereas they thrived under an iron-replete condition. The contents of chlorophyll-a, carotenoid, phycocyanin, and allophycocyanin under an iron-limited condition were lower than those under an iron-replete condition, and they all reached maximal contents on day 4 under the iron-limited condition. PS II photochemical efficiencies (maximal PS II quantum yield), saturating light levels (I-k ) and maximal electron transport rates (ETRmax) of M. aeruginosa and M. wesenbergii declined sharply under the iron-limited condition. The PS II photochemical efficiency and ETRmax of M. aeruginosa rose , whereas in the strain of M. wesenbergii, they declined gradually under the iron-replete condition. In addition, I-k of M. aeruginosa and M. wesenbergii under the iron-replete condition did not change obviously. Siderophore production of M. aeruginosa was higher than that of M. wesenbergii under the iron-limited condition. It was concluded that M. aeruginosa requires higher iron concentration for physiological and biochemical processes compared with M. wesenbergii, but its tolerance against too high a concentration of iron is weaker than M. wesenbergii.