Microcystinase(MlrA)is a key endopeptidase that catalyzes microcystin degradation without generating harmful byproduct.However,the application of MlrA in the field is primarily impeded by its limited productivity and ...Microcystinase(MlrA)is a key endopeptidase that catalyzes microcystin degradation without generating harmful byproduct.However,the application of MlrA in the field is primarily impeded by its limited productivity and short lifespan.Therefore,the MlrA's function was studied by modelling its structure,which subsequently increased its heterologous expression and high-temperature stability.Results demonstrate that after the irregular sequence at the C-terminus of MlrA was removed,enzyme solubility was significantly decreased.In addition,three fusion tags,namely maltose-binding protein,glutathione S-transferase(GST),and N-utilization substance A(NusA)were used to enhance the overexpression of soluble recombinant MlrA,among which NusA-MlrA exhibited the highest solubility.Moreover,NusA-MlrA was active in pH 4-10 at 20-80℃;even at 80℃,approximately 35.8%of fusion protein remained active.NusA-MlrA retained 89%of MlrA's activity even after 7 d of storage at 50℃;and on day 7,the protein retained>90%of its activity at pH 7.Finally,a stable,soluble,and long-lasting heterologous MlrA was successfully constructed that could eliminate microcystins in Escherichia coli C43(DE3).This study enriched the comprehension of MlrA's structure and enzymatic properties,by particularly addressing the endopeptidase's low expression and short lifespan,which improved its suitability for future applications.展开更多
Using the Widdel medium with extracted microcystin(MC)as the sole carbon and nitrogen sources,the MC-degrading bacteria community S_6 was enriched from the sediment of Litopenaeus vannamei pond,and a novel MC-degradin...Using the Widdel medium with extracted microcystin(MC)as the sole carbon and nitrogen sources,the MC-degrading bacteria community S_6 was enriched from the sediment of Litopenaeus vannamei pond,and a novel MC-degrading bacteria strain was isolated from S_6.According to 16S rDNA gene sequence and biochemical characteristics,the isolated strain was identified and named Nitratireductor aquimarinus D_(1).Fulvic acid(FA),as a widely existing photosensitizer involved in MC photodegradation,coexists with MC-degrading bacteria in natural water.The synergistic effects of N.aquimarinus D_(1) and FA on MC degradation were evaluated via comparing the degradation rate of MC induced by N.aquimarinus D_(1) and FA alone and in combination under natural light conditions.Compared with the control group,the supplementation of N.aquimarinus D_(1) and FA alone or in combination could significantly increase the degradation rate of MC(P<0.05).In the first 36 h,the degradation effect of FA on MC was better than that of N.aquimarinus D_(1),but the degradation effect was opposite at 48 h.N.aquimarinus D_(1) and FA did not show synergistic effect on MC degradation until 48 h.In the application of N.aquimarinus and FA to degrade MC in aquaculture pond,there might be a time-lag effect in the synergistic degradation.展开更多
Aeration induced turbulence was considered as an important measure to control the occurrence of cyanobacterial blooms in many lakes.Different aeration intensities were set for the culture of Microcystis aeruginosa bas...Aeration induced turbulence was considered as an important measure to control the occurrence of cyanobacterial blooms in many lakes.Different aeration intensities were set for the culture of Microcystis aeruginosa based on the formation of Microcystis colony at high iron concentrations.The turbulent dissipation rate was calculated using a computational fluid dynamics(CFD)model to evaluate the intensity of turbulence.The effects of turbulence on the formation of Microcystis colony and the release of microcystin were analyzed.Results show that turbulence produced by aeration promoted the growth of Microcystis compared to that in stagnant water.Low intensity turbulence(4×10^(-8)-1×10^(-7) m2/s3)promoted the formation of Microcystis colonies,but high intensity turbulence(1.28×10^(-6)-1.8×10^(-5) m^(2)/s^(3))did not.The increase in the number of cells per colony was slower than that in total biomass,indicating that the low intensity turbulence induced colony formation via cell division,while the high level turbulence disaggregated colonies formed by both cell division and cell adhesion.Low aeration intensity induced more production of reactive oxygen species(ROS)and malondialdehyde(MDA)in the cells of Microcystis than those in high aeration intensity.In addition,the content of microcystin(MC)-LR in the cells was positively correlated with turbulence intensity,showing that turbulence affected not only the growth and aggregation of Microcystis colonies but also their toxin production.These findings provide a better understanding of the cyanobacterial bloom formation mechanisms and help to propose feasible methods to prevent the formation of Microcystis colonies in a natural environment.展开更多
[Objective] The relationship between three isomers of microcystins and environmental factors were studied in the fields.[Method] Three isomers of microcystins （MC-LR,RR and YR） from water of five sampling spots in a...[Objective] The relationship between three isomers of microcystins and environmental factors were studied in the fields.[Method] Three isomers of microcystins （MC-LR,RR and YR） from water of five sampling spots in a northern reservoir were observed for one year with High Performance Liquid Chromatography analytical method in order to study the relationship between three isomers and environmental factors.[Result] The three isomers of microcystins showed positive correlation with chlorophyll a;LR and YR isomers all had significant linear positive correlations with the water temperature,but the RR isomer showed no significant correlation with the water temperature;LR and YR isomers had relatively significantly correlativities with the contents of total nitrogen,nitrate nitrogen and organic nitrogen,while the RR isomer only showed a significant negative correlation with the content of nitrate nitrogen;LR and RR isomers both showed significant positive correlations with the contents of total phosphorus and organic phosphorus,while the phosphorus hardly affected the YR isomer and showed no evident correlation.[Conclusion] The relationship between three isomers of microcystins and environmental factors such as chlorophyll a,water temperature,nitrogen,phosphorus were studied and investigated the reasons,which might offered a reference for controlling the growth of blue algae in water and toxin synthesis.展开更多
Lake Taihu,a large,shallow hypertrophic freshwater lake in eastern China,has experienced lake-wide toxic cyanobacterial blooms annually during summer season in the past decades.Spatial changes in the abundance of hepa...Lake Taihu,a large,shallow hypertrophic freshwater lake in eastern China,has experienced lake-wide toxic cyanobacterial blooms annually during summer season in the past decades.Spatial changes in the abundance of hepatotoxin microcystin-producing and nonmicrocystin producing Microcystis populations were investigated in the lake in August of 2009 and 2010.To monitor the densities of the total Microcystis population and the potential microcystin-producing subpopulation,we used a quantitative real-time PCR assay targeting the phycocyanin intergenic spacer（PC-IGS） and the microcystin synthetase gene（mcyD）,respectively.On the basis of quantification by real-time PCR analysis,the abundance of potential toxic Microcystis genotypes and the ratio of the mcyD subpopulation to the total Microcystis varied significantly,from 4.08×104 to 5.22×107 copies/mL,from 5.7% to 65.8%,respectively.Correlation analysis showed a strong positive relationship between chlorophyll-a,toxic Microcystis and total Microcystis;the abundance of toxic Microcystis correlated positively with total phosphorus and ortho-phosphate concentrations,but negatively with TN：TP ratio and nitrate concentrations.Meanwhile the proportion of potential toxic genotypes within Microcystis population showed positive correlation with total phosphorus and ortho-phosphate concentrations.Our data suggest that increased phosphorus loading may be a significant factor promoting the occurrence of toxic Microcystis bloom in Lake Taihu.展开更多
For purifying raw water for tap water treatment, the aquatic vegetable bed （AVB） experiment has been carded out in a hypertrophic waterfront of Taihu Lake, China. The average removal rates of total microcystin-RR an...For purifying raw water for tap water treatment, the aquatic vegetable bed （AVB） experiment has been carded out in a hypertrophic waterfront of Taihu Lake, China. The average removal rates of total microcystin-RR and microcystin-LR are 63.0% and 66. 7%, respectively. Experiments indicate that lpomoea aquatica can absorb microcystin by using enzyme-linked immunosorbent assay （ELISA）, and the roots absorb more toxins than leaves and stems. Fluorescence in situ hybridization （FISH） is used to analyze the density of microcystin degrading bacteria in the AVB sediment. Two species of microcystin degrading bacteria are detected, which indicate that microcystin bio-degradation process happened in the AVB. Protozoa and metazoa are abundant in root spheres. Aspidisca sp., Vorticella sp., Philodina sp., and Lecane sp. are dominant species. The predation functions of protozoa and metazoa have a positive effect on the removal of cyanobacteria and microcystin.展开更多
Microcystins(MCs) are cyclic hepatotoxic peptides produced by the bloom-forming cyanobacterium Microcystis and present a public health hazard to humans and livestock. The removal of MCs from contaminated water with po...Microcystins(MCs) are cyclic hepatotoxic peptides produced by the bloom-forming cyanobacterium Microcystis and present a public health hazard to humans and livestock. The removal of MCs from contaminated water with powdered activated carbon(PAC) has been employed as a simple and economic treatment strategy. In this study, PAC-Fe(Ⅲ) was prepared and utilized for the fast and efficient removal of MCs from water. PAC-Fe(Ⅲ) exhibited superior microcystin-LR(MC-LR) removal capacity and efficiency compared to the unmodified PAC. The MC-LR removal efficiency of PAC-Fe(Ⅲ) increased with decreasing p H within the pH range of 4.3 to 9.6. PAC-Fe(Ⅲ) could be reused for 3 times by methanol elution while the MC-LR removal efficiency was still over 70 percent. The removal efficiency was positively correlated to the ionic strength of water and negatively correlated to alkalinity. Natural organic matter(NOM) such as humic acid(HA) and salicylic acid(SA) generated low interference with MC-LR adsorption by PAC-Fe(Ⅲ). The complexation reaction between Fe^(3+) in PAC-Fe(Ⅲ) and the functional groups of MCLR was suggested as the key mechanism of MC-LR removal by PAC-Fe(Ⅲ). The results suggest that Femodified PAC is a promising material for the treatment of MC-contaminated waters.展开更多
[Objective] To study the differences in the tolerance of leguminous crops and cruciferous crops to microcystin （MC）. [Methed] The cruciferous typical crops oilseed rapes, pakchois, cabbages and leguminous typical cr...[Objective] To study the differences in the tolerance of leguminous crops and cruciferous crops to microcystin （MC）. [Methed] The cruciferous typical crops oilseed rapes, pakchois, cabbages and leguminous typical crops soybeans, peas and broad beans were selected as the materials to test the effects of MC of differ- ent concentrations on the germination, growth and development of leguminous crops and cruciferous crops. The measurement indicators included germination rate, plant height, chlorophyll, etc. [Result] The MC had great effects on the oilseed rape and pakchois of cruciferous crops, and smaller effects on cabbage, while the leguminous crops were generally not affected. [Conclusion] Leguminous crops are more tolerant to MC than cruciferous crops and more preferential in MC polluted regions.展开更多
The variations of environmental abundance and microcystin-LR （MC-LR） production ability of toxic Microcystis in the Nanquan region of Lake Taihu are investigated by real-time quantitative PCR （RTQ-PCR） and high pe...The variations of environmental abundance and microcystin-LR （MC-LR） production ability of toxic Microcystis in the Nanquan region of Lake Taihu are investigated by real-time quantitative PCR （RTQ-PCR） and high performance liquid chromatography （HPLC） from May to December in 2009. Simultaneously, degrees of water pollution and eutrophication are monitored. The results indicate that the water quality in the Nanquan region of Lake Taihu is in a moderate degree of pollution and eutrophication. Algal density exceeds the threshold of bloom from May to November. The environmental abundance of toxic Microcystis is more than 40% from May to October and then significantly declines to 5.66% due to the obvious reduction in the water temperature in December. From May to December, the MC-LR production ability of toxic Microcystis ranges from 1.661 to 9.293 μg/108cells. With the significant drops in water temperature and algal density, the MC-LR production ability of toxic Microcystis is obviously increased from November to December. It is concluded that the lake presents Microcystis bloom and the toxic Microcystis becomes dominant during most of the year. The environmental abundance and the MC-LR production ability of toxic Microcystis have a close relationship with water temperature. The effective control of toxic Microcystis should be considered in both the bloom period and the non-bloom period of winter since the MC-LR production ability of toxic Microcystis obviously increases in winter.展开更多
Objective To investigate the association of microcystin (MC) in drinking water with the incidence of colorectal cancer. Methods The study was designed as a retrospective cohort. Eight townships or towns were random...Objective To investigate the association of microcystin (MC) in drinking water with the incidence of colorectal cancer. Methods The study was designed as a retrospective cohort. Eight townships or towns were randomly selected as the study sites in Haining City of Zhejiang Province, China. 408 cases of colon and rectum carcinomas diagnosed from 1977 to 1996 in the study sites were included, and a survey on types of drinking water of these patients was conducted. Samples of different water sources (well, tap, river and pond) were collected separately and microcystin concentrations were determined by indirect competitive ELISA method. Results The incidence rate of colorectal cancer was significantly higher in population who drank river and pond water than those who drank well and tap water. Compared to well water, the relative risk (RR) for colorectal cancer was 1.88 (tap), 7.94 (river) and 7.70 (pond) respectively. The positive rate (>50 pg/mL) of microcystin in samples of well, tap, river and pond water was 0, 0, 36.23% and 17.14% respectively. The concentration of microcystin in river and pond water was significantly higher than that in well and tap water (P<0.01). Spearman rank correlation analysis showed that in the study sites, the microcystin concentration of river and pond water was positively associated with the incidence of colorectal cancer (rs= 0.881, P<0.01). Conclusions The types of drinking water are positively associated with the incidence of colorectal cancer in the study sites, and this may be related to microcystin contamination of drinking water. Further biological study is needed to support the possible causative role of mycrocystin in carcinogenesis of colon and rectum.展开更多
Microcystins （MCs） produced by cyanobacteria are strong hepatotoxins and classified as possible carcinogens. MCs pose a considerable threat to human health through tainted drinking and surface waters. Herein filtrat...Microcystins （MCs） produced by cyanobacteria are strong hepatotoxins and classified as possible carcinogens. MCs pose a considerable threat to human health through tainted drinking and surface waters. Herein filtrated water from a waterworks in Harbin, China, was spiked with microcysfin-LR （MC-LR） extracted from a toxic scum of microcystis aeruginosa, and the spiked sample waters were treated using UV irradiation with consequent ozonation process （UV/O3）, compared with ozonation at a dose range commonly applied in water treatment plants, UV irradiation at 254 nm and UV irradiation combined with ozonation （UV＋O3）, respectively. The remaining of toxins were analyzed using high-performance liquid chromatography and also determined using a protein phosphatase type 2A inhibition assay, which was utilized to evaluate the reduction in toxicity. Results indicated that in comparison to other three processes （O3, UV, and UV＋O3）, UV/O3 process could effectively decrease both the concentration and toxicity of MC-LR at 100 μg/L level after 5 min UV irradiation with consequent 5 min ozonation at 0.2 mg/L （below 1 μg/L ）, while 0.5 mg/L ozone dose was required for the level below 0.1 μg/L. The addition of an UV treatment step to the existing treatment train may induce significant transformation of micropollutants and breaks down the natural organic matters into moieties unfavorable for ozone decomposition, stabilizing the ozone residual. These findings suggested that sequential use of UV and ozone may be a suitable method for the removal of these potentially hazardous microcystins from drinking water.展开更多
The indigenous bacterial strain MC-LTH11 with the capability of degrading microcystin-RR MC-RR and microcystin-LR MC-LR was successfully isolated from Lake Taihu.The bacterium was identified as Stenotrophomonas sp. wh...The indigenous bacterial strain MC-LTH11 with the capability of degrading microcystin-RR MC-RR and microcystin-LR MC-LR was successfully isolated from Lake Taihu.The bacterium was identified as Stenotrophomonas sp. which possessed a mlrA gene. The MC-LTH11 thoroughly degraded MC-RR and MC-LR with the initial concentration of 37.13 mg/L and 18.49 mg /L respectively in the medium containing crude microcystins extract within 6 d.The degradation rates were affected by temperature pH initial MCs concentration and the kinds of media. Additionally the bacterial strain MC-LTH11 also degraded thoroughly microcystins in the water body of Lake Taihu within 1 d.These results suggest that the Stenotrophomonas sp.MC-LTH11 has the capacity to bioremediate water bodies contaminated by microcystins and may contribute to the degradation of microcystins after the outbreak of harmful cyanobacterial blooms in Lake Taihu.展开更多
A method using the time-resolved fluorescence technology to establish a highly sensitive microcystin-LR （MC-LR） indirect com- petitive immunoassay was proposed in this work. This method was used to monitor the MC-LR...A method using the time-resolved fluorescence technology to establish a highly sensitive microcystin-LR （MC-LR） indirect com- petitive immunoassay was proposed in this work. This method was used to monitor the MC-LR level in source water and treated drinking water from Taihu Lake. Algae in the water samples were removed by eentrifugation, and the MC-LR level was quantified using this method. Testing results showed that the sensitivity of this method was 0.01 μg/L, and the dynamic measuring range was from 0.05 to 2 μg/L. The av- erage recovery was 115%, and the variation （CV） within and between different batches were 7.3% and 9.7%, respectively. Testing results also indicated that this time-resolved fluoroimmunoassay was sensitive and accurate in measuring MC-LR level, especially for quantitative analy- sis MC-LR level in bulk water.展开更多
Seven types of microcystins,isolated from Microcystis waterbloom in Lake Dalai, were characterized.The major toxins:MCYST-LR,MCYST-RR,[D-Asp^3]MCYST-LR and [Dha^7]MCYST-LR were identified by high performance liquid ch...Seven types of microcystins,isolated from Microcystis waterbloom in Lake Dalai, were characterized.The major toxins:MCYST-LR,MCYST-RR,[D-Asp^3]MCYST-LR and [Dha^7]MCYST-LR were identified by high performance liquid chromatography(HPLC),as com- pared with the authentic microcystins.The minor toxins:MCYST FR,[L-Mser^7]MCYST-LR and an unknown MCYST which was most likely to be MCYST-(H_4)YR were identified with frit- fast atom bombardment liquid chromatography/mass spectrometry(Frit-FAB LC/MS)and amino acid analysis.The toxigenic diversity in blue-green algae(cyanobacteria)was discussed.展开更多
Harmful cyanobacterial blooms are a growing environmental problem worldwide in natural waters, the biodegradation is found to be the most efficient method for removing microcystins 0VICs） produced by harmful cyanobac...Harmful cyanobacterial blooms are a growing environmental problem worldwide in natural waters, the biodegradation is found to be the most efficient method for removing microcystins 0VICs） produced by harmful cyanobacteria. Based on the isolation of a promising bacterial strain of Sphingopyxis sp. USTB-05 for biodegrading MCs, we for the first time cloned and expressed a gene USTB-O5-A （HM245411） that is responsible for the first step in the biodegradation of microcystin LR （MC-LR） in E. coli DH5ct, with a cloning vector of pGEM-T easy and an expression vector of pGEX-4T-1, respectively. The cell-free extracts （CE） of recombinant E. coli DH5ct containing USTB-O5-A had high activity for biodegrading MC-LR. The initial MC-LR concentration of 40 mg/L was completely biodegraded within 1 hr in the presence of CE with a protein concentration of 0.35 mg/mL. Based on an analysis of the liquid chromatogram-mass spectrum （LC-MS）, the enzyme encoded by gene USTB-O5-A was found to be active in cleaving the target peptide bond between 3-amino-9-methoxy-2,6, 8-trimethyl-10-phenyl-deca-4,6-dienoic acid （Adda） and arginine of MC-LR, and converting cyclic MC-LR to linear MC-LR as a first product that is much less toxic than parent MC-LR, which offered direct evidence for the first step on the pathway of MC-LR biodegradation by Sphingopyxis sp. USTB-05.展开更多
Microcystis panniformis is a bloom forming species with flat panniform-like colonies. This species was recently found in Lake Taihu, China. To specifically characterize M. panniformis based on isolated strains, morpho...Microcystis panniformis is a bloom forming species with flat panniform-like colonies. This species was recently found in Lake Taihu, China. To specifically characterize M. panniformis based on isolated strains, morphological examination on colonial transition and genetic examination are needed. Three M. panniformis strains isolated from a water bloom sample in Lake Taihu were characterized by molecular analysis and toxin quantification. Phylogenetic analysis based on both 16S rRNA gene and internal transcribed spacer(ITS) between 16S and 23S rRNA genes were performed and compared to facilitate easy identification of the species.Relatively high similarities(98%–99%) were shown in 16S rDNA sequences between the strains of M. panniformis and those of other Microcystis species, whereas the similarities for ITS sequences were 88%–95%. In the phylogenetic tree based on the 16S rDNA sequences, the M. panniformis and M. aeruginosa strains were intermixed together with no clear division,whereas all of the M. panniformis strains were clustered together in a single clade based on the ITS sequences based phylogenyetic tree. The mcyE gene was detected in all three strains, and microcystin was determined by high-performance liquid chromatography. The molecular detection and toxin production of M. panniformis strains are of great significance for the environmental risk assessment of Microcystis blooms.展开更多
基金Supported by the National Key R&D Program of China(No.2018YFA0903100)the National Natural Science Foundation of China(No.32071601)。
文摘Microcystinase(MlrA)is a key endopeptidase that catalyzes microcystin degradation without generating harmful byproduct.However,the application of MlrA in the field is primarily impeded by its limited productivity and short lifespan.Therefore,the MlrA's function was studied by modelling its structure,which subsequently increased its heterologous expression and high-temperature stability.Results demonstrate that after the irregular sequence at the C-terminus of MlrA was removed,enzyme solubility was significantly decreased.In addition,three fusion tags,namely maltose-binding protein,glutathione S-transferase(GST),and N-utilization substance A(NusA)were used to enhance the overexpression of soluble recombinant MlrA,among which NusA-MlrA exhibited the highest solubility.Moreover,NusA-MlrA was active in pH 4-10 at 20-80℃;even at 80℃,approximately 35.8%of fusion protein remained active.NusA-MlrA retained 89%of MlrA's activity even after 7 d of storage at 50℃;and on day 7,the protein retained>90%of its activity at pH 7.Finally,a stable,soluble,and long-lasting heterologous MlrA was successfully constructed that could eliminate microcystins in Escherichia coli C43(DE3).This study enriched the comprehension of MlrA's structure and enzymatic properties,by particularly addressing the endopeptidase's low expression and short lifespan,which improved its suitability for future applications.
基金Supported by the National Natural Science Foundation of China(Nos.32172978,31772857)the Natural Science Foundation of Tianjin(No.22JCYBJC00430)+2 种基金the Tianjin Science and Technology Planning Project(No.22ZYCGSN00050)the Fundamental Research Funds of Tianjin Universities(Nos.2020ZD06,2021KJ110)the Gansu Science and Technology Project(Nos.21CX6NP223,2023ZZ1NC006)。
文摘Using the Widdel medium with extracted microcystin(MC)as the sole carbon and nitrogen sources,the MC-degrading bacteria community S_6 was enriched from the sediment of Litopenaeus vannamei pond,and a novel MC-degrading bacteria strain was isolated from S_6.According to 16S rDNA gene sequence and biochemical characteristics,the isolated strain was identified and named Nitratireductor aquimarinus D_(1).Fulvic acid(FA),as a widely existing photosensitizer involved in MC photodegradation,coexists with MC-degrading bacteria in natural water.The synergistic effects of N.aquimarinus D_(1) and FA on MC degradation were evaluated via comparing the degradation rate of MC induced by N.aquimarinus D_(1) and FA alone and in combination under natural light conditions.Compared with the control group,the supplementation of N.aquimarinus D_(1) and FA alone or in combination could significantly increase the degradation rate of MC(P<0.05).In the first 36 h,the degradation effect of FA on MC was better than that of N.aquimarinus D_(1),but the degradation effect was opposite at 48 h.N.aquimarinus D_(1) and FA did not show synergistic effect on MC degradation until 48 h.In the application of N.aquimarinus and FA to degrade MC in aquaculture pond,there might be a time-lag effect in the synergistic degradation.
基金Supported by the National Natural Science Foundation of China(No.51979236)。
文摘Aeration induced turbulence was considered as an important measure to control the occurrence of cyanobacterial blooms in many lakes.Different aeration intensities were set for the culture of Microcystis aeruginosa based on the formation of Microcystis colony at high iron concentrations.The turbulent dissipation rate was calculated using a computational fluid dynamics(CFD)model to evaluate the intensity of turbulence.The effects of turbulence on the formation of Microcystis colony and the release of microcystin were analyzed.Results show that turbulence produced by aeration promoted the growth of Microcystis compared to that in stagnant water.Low intensity turbulence(4×10^(-8)-1×10^(-7) m2/s3)promoted the formation of Microcystis colonies,but high intensity turbulence(1.28×10^(-6)-1.8×10^(-5) m^(2)/s^(3))did not.The increase in the number of cells per colony was slower than that in total biomass,indicating that the low intensity turbulence induced colony formation via cell division,while the high level turbulence disaggregated colonies formed by both cell division and cell adhesion.Low aeration intensity induced more production of reactive oxygen species(ROS)and malondialdehyde(MDA)in the cells of Microcystis than those in high aeration intensity.In addition,the content of microcystin(MC)-LR in the cells was positively correlated with turbulence intensity,showing that turbulence affected not only the growth and aggregation of Microcystis colonies but also their toxin production.These findings provide a better understanding of the cyanobacterial bloom formation mechanisms and help to propose feasible methods to prevent the formation of Microcystis colonies in a natural environment.
基金Supported by Technical Innovation Project of Ministry of Water Conservancy(SCX2002-04)National Nature Science Foundation of China(50579015)~~
文摘[Objective] The relationship between three isomers of microcystins and environmental factors were studied in the fields.[Method] Three isomers of microcystins （MC-LR,RR and YR） from water of five sampling spots in a northern reservoir were observed for one year with High Performance Liquid Chromatography analytical method in order to study the relationship between three isomers and environmental factors.[Result] The three isomers of microcystins showed positive correlation with chlorophyll a;LR and YR isomers all had significant linear positive correlations with the water temperature,but the RR isomer showed no significant correlation with the water temperature;LR and YR isomers had relatively significantly correlativities with the contents of total nitrogen,nitrate nitrogen and organic nitrogen,while the RR isomer only showed a significant negative correlation with the content of nitrate nitrogen;LR and RR isomers both showed significant positive correlations with the contents of total phosphorus and organic phosphorus,while the phosphorus hardly affected the YR isomer and showed no evident correlation.[Conclusion] The relationship between three isomers of microcystins and environmental factors such as chlorophyll a,water temperature,nitrogen,phosphorus were studied and investigated the reasons,which might offered a reference for controlling the growth of blue algae in water and toxin synthesis.
基金supported by the National Basic Research Program (973) of China (No. 2008CB418000)
文摘Lake Taihu,a large,shallow hypertrophic freshwater lake in eastern China,has experienced lake-wide toxic cyanobacterial blooms annually during summer season in the past decades.Spatial changes in the abundance of hepatotoxin microcystin-producing and nonmicrocystin producing Microcystis populations were investigated in the lake in August of 2009 and 2010.To monitor the densities of the total Microcystis population and the potential microcystin-producing subpopulation,we used a quantitative real-time PCR assay targeting the phycocyanin intergenic spacer（PC-IGS） and the microcystin synthetase gene（mcyD）,respectively.On the basis of quantification by real-time PCR analysis,the abundance of potential toxic Microcystis genotypes and the ratio of the mcyD subpopulation to the total Microcystis varied significantly,from 4.08×104 to 5.22×107 copies/mL,from 5.7% to 65.8%,respectively.Correlation analysis showed a strong positive relationship between chlorophyll-a,toxic Microcystis and total Microcystis;the abundance of toxic Microcystis correlated positively with total phosphorus and ortho-phosphate concentrations,but negatively with TN：TP ratio and nitrate concentrations.Meanwhile the proportion of potential toxic genotypes within Microcystis population showed positive correlation with total phosphorus and ortho-phosphate concentrations.Our data suggest that increased phosphorus loading may be a significant factor promoting the occurrence of toxic Microcystis bloom in Lake Taihu.
基金The National Natural Science Foundation of China(No50378014),the National High Technology Research and Develop-ment Program of China (863Program) (No2002AA601011)
文摘For purifying raw water for tap water treatment, the aquatic vegetable bed （AVB） experiment has been carded out in a hypertrophic waterfront of Taihu Lake, China. The average removal rates of total microcystin-RR and microcystin-LR are 63.0% and 66. 7%, respectively. Experiments indicate that lpomoea aquatica can absorb microcystin by using enzyme-linked immunosorbent assay （ELISA）, and the roots absorb more toxins than leaves and stems. Fluorescence in situ hybridization （FISH） is used to analyze the density of microcystin degrading bacteria in the AVB sediment. Two species of microcystin degrading bacteria are detected, which indicate that microcystin bio-degradation process happened in the AVB. Protozoa and metazoa are abundant in root spheres. Aspidisca sp., Vorticella sp., Philodina sp., and Lecane sp. are dominant species. The predation functions of protozoa and metazoa have a positive effect on the removal of cyanobacteria and microcystin.
基金Supported by the National Natural Science Foundation of China(Nos.31660138,31400405)the Jiangxi Water Science and Technology Fund(Nos.TG201501,KT201602)the Science and Technology Project from Ministry of Water Resources,China(Nos.ZXKT201508,SKY201503)
文摘Microcystins(MCs) are cyclic hepatotoxic peptides produced by the bloom-forming cyanobacterium Microcystis and present a public health hazard to humans and livestock. The removal of MCs from contaminated water with powdered activated carbon(PAC) has been employed as a simple and economic treatment strategy. In this study, PAC-Fe(Ⅲ) was prepared and utilized for the fast and efficient removal of MCs from water. PAC-Fe(Ⅲ) exhibited superior microcystin-LR(MC-LR) removal capacity and efficiency compared to the unmodified PAC. The MC-LR removal efficiency of PAC-Fe(Ⅲ) increased with decreasing p H within the pH range of 4.3 to 9.6. PAC-Fe(Ⅲ) could be reused for 3 times by methanol elution while the MC-LR removal efficiency was still over 70 percent. The removal efficiency was positively correlated to the ionic strength of water and negatively correlated to alkalinity. Natural organic matter(NOM) such as humic acid(HA) and salicylic acid(SA) generated low interference with MC-LR adsorption by PAC-Fe(Ⅲ). The complexation reaction between Fe^(3+) in PAC-Fe(Ⅲ) and the functional groups of MCLR was suggested as the key mechanism of MC-LR removal by PAC-Fe(Ⅲ). The results suggest that Femodified PAC is a promising material for the treatment of MC-contaminated waters.
基金Supported by the Key Technologies R&D Program of Henan Province(092102110105)the Docforal Foundation of Henan Institute of Engineering(D09010)+1 种基金the Critical Patented Projects in the Control and Management of National Polluted Water Bodies(2009ZX07104-005-03,2009ZX07102-003-004,2008ZX07101-007)the Open Lab Project of Hydrobiology Institutes of Chinese Academy of Sciences(2009FBZ09)~~
文摘[Objective] To study the differences in the tolerance of leguminous crops and cruciferous crops to microcystin （MC）. [Methed] The cruciferous typical crops oilseed rapes, pakchois, cabbages and leguminous typical crops soybeans, peas and broad beans were selected as the materials to test the effects of MC of differ- ent concentrations on the germination, growth and development of leguminous crops and cruciferous crops. The measurement indicators included germination rate, plant height, chlorophyll, etc. [Result] The MC had great effects on the oilseed rape and pakchois of cruciferous crops, and smaller effects on cabbage, while the leguminous crops were generally not affected. [Conclusion] Leguminous crops are more tolerant to MC than cruciferous crops and more preferential in MC polluted regions.
基金The National Key Technologies R& D Program of Chinaduring the11th Five-Year Plan Period(No.2008ZX07101-011)the National Natural Science Foundation of China(No.30972440)+1 种基金the NaturalScience Foundation of Jiangsu Province(No.BK2008320)the 333 Project Foundation of Jiangsu Province(No.07056)
文摘The variations of environmental abundance and microcystin-LR （MC-LR） production ability of toxic Microcystis in the Nanquan region of Lake Taihu are investigated by real-time quantitative PCR （RTQ-PCR） and high performance liquid chromatography （HPLC） from May to December in 2009. Simultaneously, degrees of water pollution and eutrophication are monitored. The results indicate that the water quality in the Nanquan region of Lake Taihu is in a moderate degree of pollution and eutrophication. Algal density exceeds the threshold of bloom from May to November. The environmental abundance of toxic Microcystis is more than 40% from May to October and then significantly declines to 5.66% due to the obvious reduction in the water temperature in December. From May to December, the MC-LR production ability of toxic Microcystis ranges from 1.661 to 9.293 μg/108cells. With the significant drops in water temperature and algal density, the MC-LR production ability of toxic Microcystis is obviously increased from November to December. It is concluded that the lake presents Microcystis bloom and the toxic Microcystis becomes dominant during most of the year. The environmental abundance and the MC-LR production ability of toxic Microcystis have a close relationship with water temperature. The effective control of toxic Microcystis should be considered in both the bloom period and the non-bloom period of winter since the MC-LR production ability of toxic Microcystis obviously increases in winter.
基金This project was supported by the Scientific Foundation of Ministry of Health China (No. 96-1-350)
文摘Objective To investigate the association of microcystin (MC) in drinking water with the incidence of colorectal cancer. Methods The study was designed as a retrospective cohort. Eight townships or towns were randomly selected as the study sites in Haining City of Zhejiang Province, China. 408 cases of colon and rectum carcinomas diagnosed from 1977 to 1996 in the study sites were included, and a survey on types of drinking water of these patients was conducted. Samples of different water sources (well, tap, river and pond) were collected separately and microcystin concentrations were determined by indirect competitive ELISA method. Results The incidence rate of colorectal cancer was significantly higher in population who drank river and pond water than those who drank well and tap water. Compared to well water, the relative risk (RR) for colorectal cancer was 1.88 (tap), 7.94 (river) and 7.70 (pond) respectively. The positive rate (>50 pg/mL) of microcystin in samples of well, tap, river and pond water was 0, 0, 36.23% and 17.14% respectively. The concentration of microcystin in river and pond water was significantly higher than that in well and tap water (P<0.01). Spearman rank correlation analysis showed that in the study sites, the microcystin concentration of river and pond water was positively associated with the incidence of colorectal cancer (rs= 0.881, P<0.01). Conclusions The types of drinking water are positively associated with the incidence of colorectal cancer in the study sites, and this may be related to microcystin contamination of drinking water. Further biological study is needed to support the possible causative role of mycrocystin in carcinogenesis of colon and rectum.
基金supported by the National High Technology Research and Development Program(863) of China(No. 2007AA06Z339)the 11th Five-year Plan of National Science and Technology Infrastructure Program of China(No. 2006BAJ08B02)
文摘Microcystins （MCs） produced by cyanobacteria are strong hepatotoxins and classified as possible carcinogens. MCs pose a considerable threat to human health through tainted drinking and surface waters. Herein filtrated water from a waterworks in Harbin, China, was spiked with microcysfin-LR （MC-LR） extracted from a toxic scum of microcystis aeruginosa, and the spiked sample waters were treated using UV irradiation with consequent ozonation process （UV/O3）, compared with ozonation at a dose range commonly applied in water treatment plants, UV irradiation at 254 nm and UV irradiation combined with ozonation （UV＋O3）, respectively. The remaining of toxins were analyzed using high-performance liquid chromatography and also determined using a protein phosphatase type 2A inhibition assay, which was utilized to evaluate the reduction in toxicity. Results indicated that in comparison to other three processes （O3, UV, and UV＋O3）, UV/O3 process could effectively decrease both the concentration and toxicity of MC-LR at 100 μg/L level after 5 min UV irradiation with consequent 5 min ozonation at 0.2 mg/L （below 1 μg/L ）, while 0.5 mg/L ozone dose was required for the level below 0.1 μg/L. The addition of an UV treatment step to the existing treatment train may induce significant transformation of micropollutants and breaks down the natural organic matters into moieties unfavorable for ozone decomposition, stabilizing the ozone residual. These findings suggested that sequential use of UV and ozone may be a suitable method for the removal of these potentially hazardous microcystins from drinking water.
基金The National Natural Science Foundation of China(No.30972440)the National Science and Technology Major Project(No.2012ZX07101-005)
文摘The indigenous bacterial strain MC-LTH11 with the capability of degrading microcystin-RR MC-RR and microcystin-LR MC-LR was successfully isolated from Lake Taihu.The bacterium was identified as Stenotrophomonas sp. which possessed a mlrA gene. The MC-LTH11 thoroughly degraded MC-RR and MC-LR with the initial concentration of 37.13 mg/L and 18.49 mg /L respectively in the medium containing crude microcystins extract within 6 d.The degradation rates were affected by temperature pH initial MCs concentration and the kinds of media. Additionally the bacterial strain MC-LTH11 also degraded thoroughly microcystins in the water body of Lake Taihu within 1 d.These results suggest that the Stenotrophomonas sp.MC-LTH11 has the capacity to bioremediate water bodies contaminated by microcystins and may contribute to the degradation of microcystins after the outbreak of harmful cyanobacterial blooms in Lake Taihu.
基金Project supported by Foundation of Public Health Department of Jiangsu (HD200865)National High-tech Research and Development Program (863 program) (2008AA10Z415)
文摘A method using the time-resolved fluorescence technology to establish a highly sensitive microcystin-LR （MC-LR） indirect com- petitive immunoassay was proposed in this work. This method was used to monitor the MC-LR level in source water and treated drinking water from Taihu Lake. Algae in the water samples were removed by eentrifugation, and the MC-LR level was quantified using this method. Testing results showed that the sensitivity of this method was 0.01 μg/L, and the dynamic measuring range was from 0.05 to 2 μg/L. The av- erage recovery was 115%, and the variation （CV） within and between different batches were 7.3% and 9.7%, respectively. Testing results also indicated that this time-resolved fluoroimmunoassay was sensitive and accurate in measuring MC-LR level, especially for quantitative analy- sis MC-LR level in bulk water.
文摘Seven types of microcystins,isolated from Microcystis waterbloom in Lake Dalai, were characterized.The major toxins:MCYST-LR,MCYST-RR,[D-Asp^3]MCYST-LR and [Dha^7]MCYST-LR were identified by high performance liquid chromatography(HPLC),as com- pared with the authentic microcystins.The minor toxins:MCYST FR,[L-Mser^7]MCYST-LR and an unknown MCYST which was most likely to be MCYST-(H_4)YR were identified with frit- fast atom bombardment liquid chromatography/mass spectrometry(Frit-FAB LC/MS)and amino acid analysis.The toxigenic diversity in blue-green algae(cyanobacteria)was discussed.
基金supported by the National Natural Science Foundation of China (No. 21177009)
文摘Harmful cyanobacterial blooms are a growing environmental problem worldwide in natural waters, the biodegradation is found to be the most efficient method for removing microcystins 0VICs） produced by harmful cyanobacteria. Based on the isolation of a promising bacterial strain of Sphingopyxis sp. USTB-05 for biodegrading MCs, we for the first time cloned and expressed a gene USTB-O5-A （HM245411） that is responsible for the first step in the biodegradation of microcystin LR （MC-LR） in E. coli DH5ct, with a cloning vector of pGEM-T easy and an expression vector of pGEX-4T-1, respectively. The cell-free extracts （CE） of recombinant E. coli DH5ct containing USTB-O5-A had high activity for biodegrading MC-LR. The initial MC-LR concentration of 40 mg/L was completely biodegraded within 1 hr in the presence of CE with a protein concentration of 0.35 mg/mL. Based on an analysis of the liquid chromatogram-mass spectrum （LC-MS）, the enzyme encoded by gene USTB-O5-A was found to be active in cleaving the target peptide bond between 3-amino-9-methoxy-2,6, 8-trimethyl-10-phenyl-deca-4,6-dienoic acid （Adda） and arginine of MC-LR, and converting cyclic MC-LR to linear MC-LR as a first product that is much less toxic than parent MC-LR, which offered direct evidence for the first step on the pathway of MC-LR biodegradation by Sphingopyxis sp. USTB-05.
基金supported by the National Water Science and Technology Project (No.2017ZX07203-003)the National Basic Research Program of China (No.2017YFA0605201)the support from the National Natural Science Foundation of China (No.412111020)
文摘Microcystis panniformis is a bloom forming species with flat panniform-like colonies. This species was recently found in Lake Taihu, China. To specifically characterize M. panniformis based on isolated strains, morphological examination on colonial transition and genetic examination are needed. Three M. panniformis strains isolated from a water bloom sample in Lake Taihu were characterized by molecular analysis and toxin quantification. Phylogenetic analysis based on both 16S rRNA gene and internal transcribed spacer(ITS) between 16S and 23S rRNA genes were performed and compared to facilitate easy identification of the species.Relatively high similarities(98%–99%) were shown in 16S rDNA sequences between the strains of M. panniformis and those of other Microcystis species, whereas the similarities for ITS sequences were 88%–95%. In the phylogenetic tree based on the 16S rDNA sequences, the M. panniformis and M. aeruginosa strains were intermixed together with no clear division,whereas all of the M. panniformis strains were clustered together in a single clade based on the ITS sequences based phylogenyetic tree. The mcyE gene was detected in all three strains, and microcystin was determined by high-performance liquid chromatography. The molecular detection and toxin production of M. panniformis strains are of great significance for the environmental risk assessment of Microcystis blooms.
