海河天津段微囊藻及其毒素的空间分布及与水环境因子的关系

采取荧光定量PCR技术及ELISA酶联免疫吸附法,以mcyD基因和PC-IGS基因为靶基因对夏季蓝藻暴发期间海河天津市区段的产毒微囊藻种群丰度和毒素含量进行了研究.结果表明：夏季海河天津市区段微囊藻种群丰度具有明显差异性：产毒微囊藻种群丰度为1.16×10⁴~2.48×10⁷copies/mL占总微囊藻种群的4.25%~28.59%.藻毒素含量最高点为8号采样点天津站,每升水体中藻细胞共含毒素195.51μg,除去藻细胞水中毒素浓度为0.97μg/L.总的来说,海河天津市区段微囊藻总基因拷贝数较高,产毒微囊藻丰度在不同采样点间差异较大,影响水体中微囊藻丰度的最主要环境因素是pH值.海河水体中毒素含量没有超过安全阈值,但是单位体积水体中藻细胞内毒素含量很高,具有较高的毒素释放潜力,可能对下游河口生态系统造成潜在威胁.     

微囊藻毒素的提取纯化及制备方法研究

蓝藻水华污染所带来的主要危害是由毒蓝藻向水体中释放多种不同类型的微囊藻毒素(Microcystis,MCs),其中微囊藻毒素MC-LR和MC-RR是我国富营养化水体中最主要的2种藻毒素.为解决对MCs进行深入研究中的MCs纯品缺少问题,迫切需要研究1种有效提取.纯化、制备MCs的方法.以天然水华蓝藻为原料,建立了以甲醇溶液提取、固相萃取和半制备色谱分离为主要步骤的微囊藻毒素提取、纯化和制备的方法.通过优化提取、分离和制备条件,制备了一定量的微囊藻毒素MC-LR和MC-RR高纯度样品.样品经HPLC鉴定分析,纯度可达98%以上.干燥后可得2种微囊藻毒素纯品分别为MC-RR121.1μg,MC-LR62.8μg.     

洱海水华期间饮用水源区产毒微囊藻和微囊藻毒素-LR的分布特征

水华期间微囊藻毒素的释放会严重影响洱海饮用水源水质安全.应用荧光定量PCR结合酶联免疫法快速检测洱海水华高发秋季(8—11月)主要旅游区和饮用水源地产毒微囊藻丰度和微囊藻毒素LR浓度.结果表明,荧光定量PCR方法能够快速定量洱海中总微囊藻种群和产毒微囊藻丰度,并且能有效预测洱海水体产毒潜能.洱海总微囊藻平均丰度为8.15×10~6copies·L~(-1),9月份均值最高,产毒微囊藻平均丰度为6.42×10~5copies·L~(-1),也于9月达到最高值,占总微囊藻的比例为1.0%~69.8%,水温和TP显著影响洱海总微囊藻丰度,低温、低P是洱海微囊藻水华的限制因子.洱海毒素峰值期为10月上旬,期间水源地微囊藻毒素和胞外微囊藻毒素全部检出,叶绿素a显著影响总微囊藻毒素和胞外微囊藻毒素分布,说明在一定程度上Chl a值能预测水体微囊藻释放的毒素风险.洱海总微囊藻毒素LR最高值达2.17μg·L~(-1),已超过集中式生活饮用水地表水源地对MC-LR的限值(1.0μg·L~(-1)),表明水华期间洱海饮用水水源安全问题不容忽视.     

太湖水中微囊藻毒素的测定及其分布特征

为了调查太湖水体中微囊藻毒素的污染程度,运用固相萃取-高效液相色谱技术测定了微囊藻毒素(MC-RR,MC-LR)的含量水平。该方法线性范围0.2～5 mg/L,相关系数大于0.99,2种藻毒素的最低检测限分别为0.05μg/L(MC-RR)和0.048μg/L(MC-LR)。结果表明,夏季太湖水体中MCs总体含量高于冬季;微囊藻毒素MC-LR的含量大于MC-RR。总体上看来,太湖北部(梅梁湾)水域中藻毒素的污染比其它区域水体严重。     

基于Monte Carlo模拟法对水源水体中微囊藻毒素的健康风险评估

调查水源水体中微囊藻毒素MCs(MC-RR、MC-LR和MC-YR)的污染情况,结合调查情况应用蒙特卡洛法(Monte Carlo)模拟量化人群通过饮水途径摄入微囊藻毒素的风险.在珠江西航道沿线设置5个采样点,在2016年1~6月期间共采集90份水样,根据国标(GB/T 20466-2006)推荐的HPLC方法检测水体中的微囊藻毒素,运用专业风险评估软件@Risk7.0,构建非参数概率评估模型,对通过饮水途径摄入微囊藻毒素(暴露)风险进行概率评估.对随机采集90份水源水体中微囊藻毒素MCs质量浓度检测值进行分布拟合,并运用Chi-Squared、Anderson-Darling、Kolmogorov-Smirnov这3种统计方法进行拟合度检验,根据3种评估拟合结果,确定最佳拟合分布模型.结果表明,在检测的90个水样品中,MC-RR的检出率最高,达到51.11%,质量浓度范围为0.001 7~0.386 3μg·L~(-1);其次为MC-LR和MC-YR,检出率分别是47.78%和21.11%,质量浓度范围分别是0.028 5~0.279 6μg·L~(-1)和0.003 0~0.136 2μg·L~(-1),水源水体中3种微囊藻毒素以MC-RR为主,最大检出质量浓度为0.386 3μg·L~(-1),MC-YR的含量最低.采用软件@Risk7.0分布拟合结果显示,MC-LR质量浓度最适的拟合分布为Ext Value Min模型(0.113 91,0.098 462),MC-RR质量浓度最适的拟合分布为Logistic(0.058 064,0.053 044).健康风险评估表明,MC-LR对人体健康危害的风险高于MC-RR的风险,儿童比成人更易于受到MCs污染的威胁.MC-LR对儿童健康危害的致癌年风险数值大于美国环保署(USEPA)推荐的最大可接受风险水平1×10-4;MC-LR对成人的致癌暴露年风险数值大于国际辐射防护委员会(ICRP)推荐的最大可接受风险水平5×10-5,表明水源水体中的MCs对人体健康存在潜在的危害,有必要加强饮用水源水体的保护与监控,为有效控制水源地水质污染和更好地保障人民健康奠定基础.     

巢湖夏季和冬季有毒微囊藻和无毒微囊藻种群丰度研究

应用荧光定量PCR技术研究了巢湖夏季和冬季有毒微囊藻和无毒微囊藻种群丰度的空间分布,分析了有毒微囊藻和无毒微囊藻种群丰度与环境因子之间的关系,同时应用HPLC方法分析了水体中溶解性微囊藻毒素浓度.结果表明：巢湖有毒微囊藻和无毒微囊藻种群丰度存在季节和空间差异.总的来说,夏季有毒微囊藻和无毒微囊藻种群丰度高于冬季,西湖区...     

微囊藻毒素分离及鉴定

微囊藻毒素是一类由蓝藻产生的具有肝毒性的生物毒素。微囊藻毒素标准品是开展为微囊藻毒素相关研究的必需的实验材料,文章以滇池天然水华蓝藻为原料,建立了以5%乙酸和75%的甲醇溶液提取,通过优化提取、分离和制备条件,制备了一定量的微囊藻毒素高纯度样品,经HPLC鉴定分析,纯度可达90%以上。文章在相同的质谱条件下分别对MC-LR和[Dha7]MC-LR等毒素进行了质谱检测,一级质谱结果表明,MC-LR和[Dha7]MC-LR的一价电离离子峰分别为995和981,MC-RR的二价电离离子峰为520,并分析它们的二级质谱裂解特征,确定三种毒素化学结构MC-RR为环(Ala-Arg-MeAsp-Arg-Adda-Glu-Mdha)、MC-LR为环(Ala-Leu-MeAsp-Arg-Adda-Glu-Mdha)、[Dha7]MC-LR为环(Ala-Leu-MeAsp-Arg-Adda-Glu-Dha)。改良后的检测方法同样应用于检测受微囊藻毒素污染的武汉东湖水样。     

微囊藻毒素的分离、纯化和制备

以天然水华蓝藻为原料,建立了以甲醇溶液提取、固相萃取和半制备色谱分离为主要步骤的微囊藻毒素分离纯化方法.通过优化提取、分离和制备条件,制备了一定量的微囊藻毒素MC-LR和MC-RR高纯度样品,样品经HPLC鉴定分析,纯度可达98%以上,干燥后可得2种微囊藻毒素纯品分别为MC-RR:121.1μg,MC-LR:62.8μg.     

太湖蓝藻水华期间水体和底泥中产毒微囊藻与非产毒微囊藻种群丰度研究

运用实时荧光定量PCR(Quantitative real-time PCR,QPCR)技术研究了太湖蓝藻水华期间不同湖区水体和底泥中产毒微囊藻与总微囊藻种群丰度.结果表明,湖区间产毒微囊藻种群和总微囊藻种群丰度明显不同:在水体中,竺山湾(N5)和梅梁湾(N2)产毒微囊藻和总微囊藻种群丰度高于贡湖湾(N4)和湖心(S4...     

太湖表层底泥中产毒蓝藻群落结构和种群丰度的时空变化

为了研究太湖底泥中产毒蓝藻群落结构和种群丰度的时空变化,分别于2010年冬季(2月)、春季(4月)和夏季(6月)采集太湖不同湖区底泥样品,应用基于mcyA基因的聚合酶链式反应-变性梯度凝胶电泳(PCR-DGGE)和实时定量PCR(QPCR)技术分析底泥样品.结果表明,产毒蓝藻在太湖底泥中分布广泛,其群落结构和种群丰度存在空间和季节差异性:不同湖区间群落组成相似性系数变化范围为67%～97%;冬季、春季和夏季底泥中产毒蓝藻群落结构的香农多样性指数变化范围分别为2.774～3.392、1.649～2.619和0.681～1.996,呈逐渐下降趋势;且香农多样性指数与样品中优势基因型所占比例呈极显著负相关(p<0.01,r=-0.95);从冬季到夏季,底泥中蓝藻复苏进入水体中,产毒蓝藻种群丰度逐渐降低.底泥中产毒蓝藻群落结构和种群丰度的动态变化与温度、光照、溶解氧、电导率和pH值密切相关.     

Toxic effects of acetochlor, methamidophos and their combination on nifH gene in soil

Toxic effects of two agrochemicals on nifH gene in agricultural black soil were investigated using denaturing gradient gel electrophoresis （DGGE） and sequencing approaches in a microcosm experiment. Changes of soil nifH gene diversity and composition were examined following the application of acetochlor, methamidophos and their combination. Acetochlor reduced the nifH gene diversity （both in gene richness and diversity index values） and caused changes in the nifH gene composition. The effects of acetochlor on nifH gene were strengthened as the concentration of acetochlor increased. Cluster analysis of DGGE banding patterns showed that nifH gene composition which had been affected by low concentration of acetochlor （50 mg/kg） recovered firstly. Methamidophos reduced nifH gene richness that except at 4 weeks. The medium concentration of methamidophos （150 mg/kg） caused the most apparent changes in nifH gene diversity at the first week while the high concentration of methamidophos （250 mg/kg） produced prominent effects on nifH gene diversity in the following weeks. Cluster analysis showed that minimal changes of nifH gene composition were found at 1 week and maximal changes at 4 weeks. Toxic effects of acetochlor and methamidophos combination on nifH gene were also apparent. Different nifH genes （bands） responded differently to the impact of agrochemicals： four individual bands were eliminated by the application of the agrochemicals, five bands became predominant by the stimulation of the agrochemicals, and four bands showed strong resistance to the influence of the agrochemicals. Fifteen prominent bands were partially sequenced, yielding 15 different nifH sequences, which were used for phylogenetic reconstructions. All sequences were affiliated with the alpha- and beta-proteobacteria, showing higher similarity to eight different diazotrophic genera.     

EDTA-enhanced phytoremediation of lead contaminated soil by Bidens maximowicziana

Phytoremediation is a potential cleanup technology for the removal of heavy metals from contaminated soils.Bidens maximowicziana is a new Pb hyperaccumulator,which not only has remarkable tolerance to Pb but also extraordinary accumulation capacity for Pb.The maximum Pb concentration was 1509.3 mg/kg in roots and 2164.7 mg/kg in overground tissues.The Pb distribution order in the B. maximowicziana was:leaf>stem>root.The effect of amendments on phytoremediation was also studied.The mobility of soil Pb and the Pb concentrations in plants were both increased by EDTA application.Compared with CK(control check),EDTA application promoted translocation of Pb to overground parts of the plant.The Pb concentrations in overground parts of plants was increased from 24.23-680.56 mg/kg to 29.07-1905.57 mg/kg.This research demonstrated that B.maximowicziana appeared to be suitable for phytoremediation of Pb contaminated soil,especially,combination with EDTA.     

Distribution and ecological effect of mercury in Laogang landfill, Shanghai, China

Laogang landfill near Shanghai is the largest landfill in China, and receives about 10000 t of daily garbage per day, Samples of topsoil and plants were analyzed to evaluate mercury pollution from the landfill. For topsoil samples, there were significant correlations among total mercury （HgT）, combinative mercury （Hgc） and gaseous mercury （HgG）, and content of total organic carbon （TOC）, but, no significantly relationship was found between Hg content and filling time. Hg content changes in vertical profiles with time showed that the average Hgv of profiles 1992, 1996, and 2000 was similar, but their average HgG was quite different. HgT was significantly correlated with Hgc in profile 1992 and 2000, and Hgv was significantly correlated with Hg6 in profile 1996. HgG/Hgv ratio in profile samples decreased in the order of （HgG,/HgT）1992〉（HgG/HgT）1996〉〉（HgG/HgT）2000. A simple outline of Hg release in landfill could be drawn： with increasing of filling time, degradation undergoes different biodegradation, accordingly, gaseous mercury goes through small, more, and small proportion to total mercury. Distribution of Hg in plants was inhomogeneous, following the order of leaf〉root〉stem. The highest value of leaf may be associated with higher atmospheric Hg from landfill. Ligneous plants （e.g. Phyllostachys glanca, Prunus salicina and Ligustrum lucidum） are capable of enriching more Hg than herbaceous plants.     

Arsenic uptake by arbuscular mycorrhizal maize （Zea mays L.） grown in an arsenic-contaminated soil with added phosphorus

The effects of arbuscular mycorrhizal (AM) fungus (Glomus mosseae) and phosphorus (P) addition (100 mg/kg soil) on arsenic (As) uptake by maize plants (Zea mays L.) from an As-contaminated soil were examined in a glasshouse experiment.Non-mycorrhizal and zero-P addition controls were included.Plant biomass and concentrations and uptake of As,P,and other nutrients,AM colonization,root lengths,and hyphal length densities were determined.The results indicated that addition of P significantly inhibited root colonization and development of extraradical mycelium.Root length and dry weight both increased markedly with mycorrhizal colonization under the zero-P treatments,but shoot and root biomass of AM plants was depressed by P application.AM fungal inoculation decreased shoot As concentrations when no P was added,and shoot and root As concentrations of AM plants increased 2.6 and 1.4 times with P addition,respectively.Shoot and root uptake of P,Mn,Cu,and Zn increased,but shoot Fe uptake decreased by 44.6%,with inoculation, when P was added.P addition reduced shoot P,Fe,Mn,Cu,and Zn uptake of AM plants,but increased root Fe and Mn uptake of the nonmycorrhizal ones.AM colonization therefore appeared to enhance plant tolerance to As in low P soil,and have some potential for the phytostabilization of As-contaminated soil,however,P application may introduce additional environmental risk by increasing soil As mobility.     

Enzymatic oxidation of aqueous pentachlorophenol

The influence of the nonionic surfactant Tween 80 on pentachlorophenol （PCP） oxidation catalyzed by horseradish peroxidase was studied. The surfactant was tested at concentrations below and above its critical micelle concentration （CMC）. Enhancement of PCP removal was observed at sub-CMCs. The presence of Tween 80 in the reaction mixture reduced enzyme inactivation which occurred through a combination of free radical attack and sorption by precipitated products. A simple first-order model was able to simulate time profiles for enzyme inactivation in the presence or absence of Tween 80. At supra-CMCs, the surfactant caused noticeable reductions in PCP removal, presumably through micelle partitioning of PCP which precluded the hydrophobic PCP molecule from interacting with the enzyme.     

Decomposition of alachlor by ozonation and its mechanism

Decomposition and corresponding mechanism of alachlor, an endocrine disruptor in water by ozonation were investigated. Results showed that alachlor could not be completely mineralized by ozone alone. Many intermediates and final products were formed during the process, including aromatic compounds, aliphatic carboxylic acids, and inorganic ions. In evoluting these products, some of them with weak polarity were qualitatively identified by GC-MS. The information of inorganic ions suggested that the dechlorination was the first and the fastest step in the ozonation of alachlor.     

Degradation of metabolites accumulated of benzo[a]pyrene by coupling Penicillium Chrysogenum with KMnO4

Several main metabolites of benzo[a]pyrene （BaP） formed by Penicillium chrysogenum, Benzo[a]pyrene-1,6-quinone （BP 1,6- quinone）, trans-7,8-dihydroxy-7,8-dihydrobenzo[a]pyrene （BP 7,8-diol）, 3-hydroxybenzo[a]pyrene （3-OHBP）, were identified by high-performance liquid chromatography （HPLC）. The three metabolites were liable to be accumulated and were hardly further metabolized because of their toxicity to microorganisms. However, their further degradation was essential for the complete degradation of BaP. To enhance their degradation, two methods, degradation by coupling Penicillium chrysogenum with KMnO4 and degradation only by Penicillium chrysogenum, were compared; Meanwhile, the parameters of degradation in the superior method were optimized. The results showed that （1） the method of coupling Penicillium chrysogenum with KMnO4 was better and was the first method to be used in the degradation of BaP and its metabolites; （2） the metabolite, BP 1,6-quinone was the most liable to be accumulated in pure cultures; （3） the effect of degradation was the best when the concentration of KMnO4 in the cultures was 0.01% （w/v）, concentration of the three compounds was 5 mg/L and pH was 6.2. Based on the experimental results, a novel concept with regard to the bioremediation of BaP-contaminated environment was discussed, considering the influence on environmental toxicity of the accumulated metabolites.     

Impact of atrazine and nitrogen fertilizers on the sorption of chlorotoluron in soil and model sorbents

Sorption of chlorotoluron in ammonium sulfate, urea and atrazine multi-solutes system was investigated by batch experiments. The results showed application of nitrogen fertilizers to the soil could affect the behavior of chlorotoluron. At the same concentration of N, sorption of chlorotoluron decreased as the concentration of atrazine increased on the day 0 and 6 in soil, respectively. The sorption of chlorotoluron increased from 0 to 6 d when soils were preincubated with deionized water, ammonium sulfate and urea solution for 6 d. That indicated incubation time was one of the most important factors for the sorption of chlorotoluron in nitrogen fertilizers treatments. The individual sorption isotherms of chlorotoluron in rubbery polymer and silica were strictly linear in single solute system, but there were competition sorption between pesticides or between pesticides and nitrogen fertilizers. That indicated the sorption taken place by concurrent solid-phase dissolution mechanism and sorption on the interface of water-organic matter or water-mineral matter.     

基于综合水质标识指数和对应分析法的校园河道水质评价

以三峡大学的校园河道求索溪为研究对象,利用综合水质标识指数法确定求索溪水质类别,分析其水质时空变化规律,并利用对应分析法得出求索溪中不同监测点的主要污染因子.研究结果表明:求索溪整体的综合水质标识指数为7.423,整体水质为劣V类(地表水环境质量标准GB 3838-2002)且黑臭.从时间变化来看,求索溪4月份的水质最差,5月份次之,4、5月份所有监测点的水质都劣于V类且黑臭;8月份水质最好,水质为Ⅳ类;从空间分布来看,8个监测点综合水质标识指数均超过6.0,水质为劣V类,其中6号监测点的水质相对最好,监测点3号的水质相对最差;对应分析法得出求索溪的整体水体污染程度受总氮因子的影响最大,其次为总磷.该研究拟为求索溪及类似校园河道的水环境治理研究提供基础依据和参考.     

Potential of plant polyphenol oxidases in the decolorization and removal of textile and non-textile dyes

In this study an effort has been made to use plant polyphenol oxidases; potato （Solanum tuberosum） and brinjal （Solanum melongena）, for the treatment of various important dyes used in textile and other industries. The ammonium sulphate fractionated enzyme preparations were used to treat a number of dyes under various experimental conditions. Majority of the treated dyes were maximally decolorized at pH 3.0. Some of the dyes were quickly decolorized whereas others were marginally decolorized. The initial first hour was sufficient for the maximum decolorization of dyes. The rate of decolorization was quite slow on long treatment of dyes. Enhancement in the dye decolorization was noticed on increasing the concentration of enzymes. The complex mixtures of dyes were treated with both preparations of polyphenol oxidases in the buffers of varying pH values. Potato polyphenol oxidase was significantly more effective in decolorizing the dyes to higher extent as compared to the enzyme obtained from brinjal polyphenol oxidase. Decolorization of dyes and their mixtures, followed by the formation of an insoluble precipitate, which could be easily removed simply by centrifugation.