Lirong Song

Responses of antioxidant systems in the hepatocytes of common carp (Cyprinus carpio L.) to the toxicity of microcystin-LR

The freshwater, bloom-forming cyanobacterium (blue-green alga) Microcystis aeruginosa produces a peptide hepatotoxin, which causes the damage of animal liver. Recently, toxic Microcystis blooms frequently occur in the eutrophic Dianchi Lake (300 km2 and located in the South-Western of China). Microcystin-LR from Microcystis in Dianchi was isolated and purified by high performance liquid chromatography (HPLC) and its toxicity to mouse and fish liver was studied (Li et al., 2001). In this study, six biochemical parameters (reactive oxygen species, glutathione, superoxide dismutase, catalase, glutathione peroxide and glutathione S-transferase) were determined in common carp hepatocytes when the cells were exposed to 10 microg microcystin-LR per litre. The results showed that reactive oxygen species (ROS) contents increased by more than one-time compared with the control after 6 h exposure to the toxin. In contrast, glutathione (GSH) levels in the hepatocytes exposed to microcystin-LR decreased by 47% compared with the control. The activities of superoxide dismutase (SOD), catalase (CAT) and glutathione peroxide (GSH-Px) increased significantly after 6 h exposure to microcystin-LR, but glutathione S-transferase (GST) activity showed no difference from the control. These results suggested that the toxicity of microcystin-LR caused the increase of ROS contents and the depletion of GSH in hepatocytes exposed to the toxin and these changes led to oxidant shock in hepatocytes. Increases of SOD, CAT and GSH-Px activities revealed that these three kinds of antioxidant enzymes might play important roles in eliminating the excessive ROS. This paper also examined the possible toxicity mechanism of microcystin-LR on the fish hepatocytes and the results were similar to those with mouse hepatocytes.

Effect of desert soil algae on the stabilization of fine sands

Four filamentous cyanobacteria, Microcoleusvaginatus, Phormidium tenue,Scytonemajavanicum (Kutz.) and Nostoc sp., and asingle-celled green alga, Desmococcus olivaceus, allisolated from Shapotou (Ningxia Hui Autonomous Region of China), were batchcultured and inoculated onto unconsolidated sand in greenhouse and fieldexperiments. Their ability to reduce wind erosion in sands was quantified byusing a wind tunnel laboratory. The major factors related to cohesion of algalcrusts, such as biomass, species, species combinations, bioactivity, niche,growth phase of algae, moisture, thickness of the crusts, dust accretion(including dust content and manner of dust added) and other cryptogams(lichens,fungi and mosses) were studied. The best of the five species were M.vaginatus and P. tenue, while the best mix wasablend of 80% M. vaginatus and 5% each of P.tenue,S. javanicum,Nostocsp. and D. olivaceus. The threshold friction velocity wassignificantly increased by the presence of all of the cyanobacterial species,while the threshold impact velocity was notably increased only by thefilamentous species. Thick crusts were less easily eroded than thin crusts,while biomass was more effective than thickness. Dust was incorporated bestintoMicrocoleus crust when added in small amounts over time,and appeared to increase growth of the cyanobacterium as well as strengthen thecohesion of the crust. Microbial crust cohesion was mainly attributed to algalaggregation, while lichens, fungi and mosses affected more the soil structureand physico-chemical properties.

The toxic effects of microcystin-LR on embryo-larval and juvenile development of loach, Misguruns mizolepis Gunthe

Microcystin-LR, a specific and potent hepatotoxin, was tested for its effects on loach embryo-larval and juvenile development. The results of this study showed that loach embryos were more sensitive when exposed to microcystin-LR at a later than at an earlier stage of development. Juveniles were far less sensitive to MC-LR than were embryos and larvae. Mortality and developmental abnormality were proven to be dose-dependent and to be stage-specific sensitive. Among the abnormal changes noted were: pericardial edema and tubular heart, bradycardia, homeostasis, poor yolk resumption, small head, curved body and tail, and abnormal hatching. Liver and heart were the main targets of microcystin-LR toxicity. Ultrastructural analysis documented a complex set of sublethal effects of microcystin-LR on loach hepatocytes, chiefly including morphological alteration in nuclear and RER of loach liver cells. In addition, microcystin-LR was lethal to loach juvenile in the subacute (7 days) exposure (LC(50)=593.3 microg/l).

The role of microcystins in maintaining colonies of bloom-forming Microcystis spp.

Microcystis is a cosmopolitan genus of cyanobacteria and occurs in many different forms. Large surface blooms of the cyanobacterium are well known in eutrophic lakes throughout the globe. We evaluated the role of microcystins (MCs) in promoting and maintaining bloom-forming cell aggregates at environmentally relevant MC concentrations (0.25-10 µg l(-1)). MCs significantly enhanced Microcystis colony sizes. Colonial diameters in microcystin-RR (MC-RR)-treated cultures (at 1 µg l(-1)) were significantly larger than control colonies, by factors of 1.5, 2.6 and 2.7 in Microcystis wesenbergii DC-M1, M. ichthyoblabe TH-M1 and Microcystis sp. FACHB1027 respectively. Depletion of extracellular MC concentrations caused Microcystis colony size to decrease, suggesting that released MCs are intimately involved in the maintenance of Microcystis colonial size. MC-RR exposure did not influence Microcystis growth rate, but did significantly increase the production of extracellular polysaccharides (EPS). In addition, MC-RR exposure appeared to trigger upregulation of certain parts of four polysaccharide biosynthesis-related genes: capD, csaB, tagH and epsL. These results strongly indicate that induction of polysaccharides by MC-RR was the major mechanism through which MCs enhanced colony formation in Microcystis spp. Cellular release of MCs, therefore, may play a key role in the persistence of algal colonies and the dominance of Microcystis.

Microcystin production of Microcystis viridis (cyanobacteria) under different culture conditions

Toxic cyanobacterium Microcystis viridis was isolated from the eutrophic Dianchi Lake of south‐western China. Three microcystins were detected from this strain by high‐pressure liquid chromatography. Among them, [Dha7] microcystin‐RR is the major component and represents about 70–80% of the total toxins of this strain. We examined its microcystin production pattern in relation to culture conditions such as light intensity, nutrition, temperature, pH and growth phase. Of all the factors examined, light intensity and pH influenced the toxin production much more than other factors. At 25°C, a high production of [Dha7] microcystin‐RR was obtained at light intensity 15 μE s‐1 m2, and both low and high pH (pH 7.0 and pH 9.2) enhanced the yield of microcystin‐RR. No strong relationships were found between total toxin and temperature ranging from 15°C. The highest production of total toxins was measured in the mid‐exponential growth phase and lowest production towards the declining growth phase. Remarkable changes in toxin production and morphology emerged coincidentally following the transfer of nitrogen‐deficient cells into fresh MA medium during the time course. These findings may shed light on the correlation between toxin production and morphogenesis of this strain.

Comparative studies on physiological responses to phosphorus in two phenotypes of bloom-forming Microcystis

Toxic Microcystis blooms frequently occur in eutrophic water bodies and exist in the form of colonial and unicellular cells. In order to understand the mechanism of Microcystis dominance in freshwater bodies, the physiological and biochemical responses of unicellular (4 strains) and colonial (4 strains) Microcystis strains to phosphorus (P) were comparatively studied. The two phenotype strains exhibit physiological differences mainly in terms of their response to low P concentrations. The growth of four unicellular and one small colonial Microcystis strain was significantly inhibited at a P concentration of 0.2 mg l−1; however, that of the large colonial Microcystis strains was not inhibited. The results of phosphate uptake experiments conducted using P-starved cells indicated that the colonial strains had a higher affinity for low levels of P. The unicellular strains consumed more P than the colonial strains. Alkaline phosphatase activity in the unicellular strains was significantly induced by low P concentrations. Under P-limited conditions, the oxygen evolution rate, Fv/Fm, and ETRmax were lower in unicellular strains than in colonial strains. These findings may shed light on the mechanism by which colonial Microcystis strains have an advantage with regard to dominance and persistence in fluctuating P conditions.

Health risks associated with consumption of microcystin-contaminated fish and shellfish in three Chinese lakes: significance for freshwater aquacultures.

The risks associated with consuming aquatic products were systematically evaluated by analyzing 26 economically important fish and shellfish species which were harvested monthly from three large lakes in China during the fisheries catch season. Results indicate most of the aquatic products from the three large lakes seem to be unsafe for human consumption due to microcystin accumulations, with the estimated daily intake (EDI) values 5-148 times, 2-50 times and 1.5-4 times higher than the tolerable daily intake (TDI) value in Taihu, Chaohu and Dianchi, respectively. In addition, the toxin accumulation in the harvested organisms varied intensity from month to month and by species which suggests that consumption risks may be reduced or avoided by either adjusting the legal fishing seasons or the species of fish and shellfish harvested. This study will provide new information about the risks associated with the consumption of aquatic products and suggests possible management strategies to reduce or avoid potential health risks.

Activation of Nrf2 by Microcystin-LR Provides Advantages for Liver Cancer Cell Growth

Microcystin-LR (MC-LR) is a potent heptapeptide hepatotoxin at high doses, but its underlying mechanism of promoting liver cell proliferation at low doses is unclear. The transcription factor nuclear factor erythroid 2-related factor 2 (Nrf2) is key in mediating the protective antioxidant response against various environmental toxicants, but emerging data suggest that constitutive activation of Nrf2 contributes to a malignant phenotype. The purpose of this study was to characterize the interactions and effects of Nrf2 activation on cell proliferation induced by MC-LR treatment. Treatment of HepG2 and Hep3B cells with MC-LR resulted in significant increases in Nrf2-ARE binding activities in the nuclear fractions and upregulation of its downstream genes HO-1 and NQO1. A possible mechanism may be that MC-LR binds to the cytosolic regulator protein Keap1 to liberate Nrf2. Nrf2 knockdown inhibited MC-LR-induced cell proliferation and cell cycle progression. Together, these results indicate that MC-LR-induced upregulation of Nrf2 in cancer cells promotes liver cancer cell growth and suggest a positive role of Nrf2 in tumorigenesis.

Physiological Comparison between Colonial and Unicellular forms of Microcystis Aeruginosa Kütz. (Cyanobacteria)

Z-X. Wu and L-R. Song. 2007. Physiological comparison between colonial and unicellular forms of Microcystis aeruginosa Kütz. (Cyanobacteria). Phycologia 47: 98–104. DOI: 10.2216/07–36.1 In order to gain insight into the bloom sustainment of colonial Microcystis aeruginosa Kütz., physiological characterizations were undertaken in this study. Compared with unicellular Microcystis, colonial Microcystis phenotypes exhibited a higher maximum photosynthetic rate (Pm), a higher maximum electron transfer rate (ETRmax), higher phycocyanin content, and a higher affinity for inorganic carbon (K0.5 DIC ≤ 8.4 ± 0.7 µM) during the growth period monitored in this study. This suggests that photosynthetic efficiency is a dominant physiological adaptation found in colonial Microcystis, thus promoting bloom sustainment. In addition, the high content of soluble and total carbohydrates in colonial Microcystis suggests that this phenotype may possess a higher ability to tolerate enhanced stress conditions when compared to unicellular (noncolonial) phenotypes. Therefore, high photosynthetic activities and high tolerance abilities may explain the bloom sustainment of colonial Microcystis in eutrophic lakes.

Gene expression profiles in liver of zebrafish treated with microcystin-LR

Microcystin-LR (MC-LR) is the most frequently studied cyclic heptatoxin produced by cyanobacteria, which has tremendous negative impacts on fish, while its molecular mechanism behind remained unclear at present. Here, Affymetrix Zebrafish GeneChip was used to identify alterations in gene expression of zebrafish (Danio rerio) after MC-LR exposure. Among the 14,900 transcripts in the microarray, 273 genes were differentially expressed, in which 243 genes were elevated and 30 were decreased. According to GOstat analysis, MC-LR mainly influenced the cell cycle and mitogen-activated protein kinases (MAPK) signaling pathways. In addition, many immune-related genes were also influenced. These data suggest that MC-LR could promote tumorigenesis and cause immunotoxicity in fish.