恩诺沙星完全抗原的合成及间接竞争ELISA方法的建立

为了建立环境新型抗生素类污染物恩诺沙星的免疫检测方法,采用间接竞争ELISA方法,用酶标仪测定酶标二抗与底物反应生成的有色产物量来确定恩诺沙星含量.研究了恩诺沙星完全抗原的合成方法,以及间接竞争ELISA的优化条件和实际水样的检测效果.结果表明,通过碳二亚胺法(EDC+NHS)成功合成了恩诺沙星的理想完全抗原,采用BCA法测定完全抗原浓度,紫外吸收法和MADLI-TOF MS法测得完全抗原的偶联比为14~16.建立的恩诺沙星间接竞争ELISA检测方法,检测限为1.92μg·L~(-1),定量检测区间为7.33~238.24μg·L~(-1).采用优化后的实验条件测定了清华大学地下水和饮用水配制的恩诺沙星加标样品,发现所有样品的变异系数和回收率分别为3%~11%和82%~122%,表明建立的间接竞争ELISA方法适用于实际水样中恩诺沙星的快速简便检测.     

城市生活垃圾填埋场中抗生素残留特征研究

为研究城市生活垃圾填埋场中抗生素的残留及其浓度特征,采集上海市老港填埋场中固体废弃物和渗滤液样本,采用超高效液相色谱三重四级杆质谱联用技术(UPLC-3Q-MS)检测抗生素浓度.结果显示,固体废弃物和渗滤液中四环素类、磺胺类、喹诺酮类、大环内脂类及β-内酰胺类抗生素的检出浓度范围分别为0.1~4.9μg·kg~(-1),nd~3245.0 ng·L~(-1)、nd~16.3μg·kg~(-1),nd~1360.0 ng·L~(-1)、0.2~367.5μg·kg~(-1),nd~6950.0ng·L~(-1)、LOQ~8.4μg·kg~(-1),0.3~9450.0 ng·L~(-1)及nd~2.8μg·kg~(-1),nd~293.5 ng·L~(-1).抗生素浓度在垃圾填埋场中的分布存在时空差异,填埋场1.5 m处固体废弃物中抗生素的浓度高于0.5 m处,新鲜渗滤液中抗生素浓度大于老龄渗滤液.研究结果为可今后垃圾填埋场中残留抗生素的治理提供理论依据.     

环境中典型抗生素生态安全阈值

随着人们生活水平的提高,抗生素的使用量日趋增加,对生态环境造成潜在风险.以12种典型抗生素(克拉霉素、金霉素、青霉素、头孢噻肟、头孢唑啉、硫酸链霉素、新霉素、磺胺甲恶唑、林可霉素、氯霉素、诺氟沙星、恩诺沙星)为研究对象,根据欧盟化学物质风险评价技术指导文件(TGD),推导不同环境介质中抗生素的预测无效应浓度(PNEC).结果表明,12种典型抗生素PNECwater值大小依次为：磺胺甲恶唑(0.0075μg·L^-1)<青霉素(0.10μg·L^-1)<诺氟沙星(0.1128μg·L^-1)<克拉霉素(0.20μg·L^-1)<金霉素(0.24μg·L^-1)<新霉素(0.60μg·L^-1)<硫酸链霉素(1.00μg·L^-1)<林可霉素(2.00μg·L^-1)<氯霉素(12.5μg·L^-1)=恩诺沙星(12.5μg·L^-1)<头...     

胶网藻对水体中恩诺沙星的毒性响应及去除作用

恩诺沙星是医药及水产养殖业广泛使用的喹诺酮类抗生素,是我国地表水中残留量较多的抗生素种类之一;然而有关该抗生素对水体中浮游植物毒理作用以及后者对其去除作用的研究还鲜见报道.本研究以绿藻门的胶网藻(Dictyosphaerium sp.)为对象,通过12 d室内培养实验,分析在不同浓度(0、 5、 25、 50和100 mg·L~(-1))恩诺沙星暴露的条件下,胶网藻的生长、光合作用活力和胞外多糖的响应变化及其对不同浓度恩诺沙星的去除效率.结果表明,各处理组中胶网藻的生物量和光合色素含量均随培养时间的增加而升高,但与对照组相比,恩诺沙星对胶网藻的生长和光合色素积累均产生显著抑制作用(P0.01).通过拟合分析,计算出恩诺沙星对胶网藻96 h半数致死浓度(LC_(50))为(241.29±7.33) mg·L~(-1),表明胶网藻能够适应高浓度的恩诺沙星胁迫.同时发现,当恩诺沙星浓度5 mg·L~(-1)时,能够促进胶网藻的最大光合速率(F_v/F_m),但浓度5 mg·L~(-1)时则对胶网藻具有抑制作用(P0.01);实际光合速率(Yeild)和最大电子传递速率(ETR_(max))在12 d内呈现先降后升的变化趋势,胶网藻在6 d后可逐渐适应胁迫环境而恢复一定的光合活力.此外,恩诺沙星可刺激胶网藻释放胞外多糖,水溶性多糖(RPS)和胞外胶鞘多糖(CPS)含量均随恩诺沙星浓度升高而增加.实验结束时, 4个无藻对照组(5、 25、 50和100 mg·L~(-1))中恩诺沙星的去除率分别为7.27%、 5.56%、 5.30%和4.88%,而添加胶网藻的处理组其去除率分别为对照组的3.21、 3.01、 2.69和2.83倍,表明胶网藻对于恩诺沙星的去除具有显著的促进作用(P0.01).本研究结果为理解喹诺酮类抗生素对水体初级生产者的生态毒性提供了新资料,同时也为水体抗生素残留的生态去除和淡水微藻的生物资源化利用提供了新的思路.     

基于叶绿素荧光成像技术的藻毒性检测法的建立及在环境监测中的应用

应用新型大面积调制叶绿素荧光成像系统(Maxi-Imaging-PAM),以蛋白核小球藻作为指示生物,以典型的光合作用抑制剂敌草隆和莠灭净为测试物质,建立了微孔板式藻毒性检测方法,确定了实验参数对结果的影响,并将该方法应用于检测石油废水的藻毒性.结果表明,最适实验条件为:藻细胞的暴露浓度在3.45×10~6～5.28×10~6 cell·mL~(-1)范围内,暴露时间为2 h时,毒性检测最灵敏,获得的敌草隆和莠灭净的半最大效应浓度(EC_(50))分别为(4.33±0.38)μg·L~(-1)和(3.87±0.27)μg·L~(-1).将该方法成功应用于某石油废水的藻毒性检测,确定其EC_(50)值为1.03%±0.07%(稀释百分数),相当于414.96μg·L~(-1)的敌草隆的毒性.本研究建立的微孔板式藻毒性检测方法具有灵敏度高、所需样品量少、可大批量测试等优点,对毒理学发展具有重要意义.     

莲花水库水体中抗生素污染特征及生态风险评价

采用固相萃取-高效液相色谱串联质谱法对厦门市新建饮用水源地莲花水库中4类(四环素类、喹诺酮类、大环内酯类和磺胺类)13种典型抗生素进行了检测,并评价了其污染特征和生态风险等级.结果表明,除红霉素、磺胺二甲嘧啶和磺胺甲■唑外,其余10种抗生素均有不同程度检出,总浓度范围为n.d.～925.26 ng·L~(-1).其中阿奇霉素的浓度最高(n.d.～232.61 ng·L~(-1)),检出率为75%;其次为恩诺沙星(n.d.～187.69ng·L~(-1))、四环素(n.d.～155.05 ng·L~(-1))和环丙沙星(n.d.～83.66 ng·L~(-1)),检出率均超过60%.抗生素浓度随采样点呈现出上游莲花溪S1澳溪支流S2库区下游S3入库口S4库区中心S5的趋势.抗生素季节分布特征较为明显,枯水期总浓度明显高于丰水期和平水期.生态环境风险评价表明氧氟沙星、恩诺沙星和环丙沙星的生态环境风险较高,环丙沙星为主要风险因子;枯水期的抗生素联合风险商值比丰水期和平水期高,且大于1,对生态环境存在较高风险,应引起相关部门足够重视.     

天津市规模化奶牛养殖场废水中典型抗生素处理效果及生态风险评估

为了解天津市规模化奶牛养殖场废水处理前后典型抗生素的浓度水平及处理效果,使用固相萃取-超高效液相色谱-串联三重四极杆质谱仪(SPE-UPLC-MS/MS)技术,对天津市12家规模化奶牛养殖场废水中7种典型抗生素进行调查监测.结果表明,12家规模化奶牛养殖场废水处理前后均不同程度地检测出抗生素残留.废水处理前,替米考星(TIL)、盐酸土霉素(OTC)、司帕沙星(SPA)、磺胺噻唑(STZ)、氧氟沙星(OFL)和盐酸沙拉沙星(SAR)检出率均为100%,磺胺嘧啶(SDZ)检出率为83. 33%.其中替米考星和盐酸土霉素为未处理废水中主要的抗生素成分,浓度为25. 21μg·L~(-1)和9. 87μg·L~(-1).处理后废水中磺胺嘧啶和氧氟沙星的检出率下降至25. 00%和41. 66%,主要成分为替米考星和盐酸土霉素,但浓度有着明显地降低,分别为11. 30μg·L~(-1)和3. 71μg·L~(-1).不同的规模化奶牛养殖场对于抗生素的处理效果在24. 95%～81. 05%之间.厌氧-缺氧-好氧(AAO)处理工艺的综合处理效果优于厌氧好氧(AO)处理工艺.氧氟沙星、盐酸沙拉沙星、盐酸土霉素为处理后废水中主要的高风险污染物,各规模化奶牛养殖场都含有一种或多种RQs 1的抗生素,其排放对环境构成一定的生态风险.     

基于稀疏表达的水体遥感反射率高光谱重构及其应用

高光谱重构技术可以有效地突破多光谱卫星传感器波段设置的限制,获得更多更有效的地物光谱信息.本研究基于稀疏表达方法提出了一种针对水体遥感反射率的高光谱重构算法,以太湖、杭州湾的原位水体光谱数据为数据源,在5种常用水色传感器(Sentinel-2A MSI、MERIS、MODIS Aqua、GOCI以及ⅦRS)上进行了高光谱重构实验,最后将该算法应用于GOCI数据,进行了算法适用性验证.结果表明:(1)基于稀疏表达的高光谱重构算法可以在不利用实测光谱数据的情况下实现高光谱重构,光谱重构精度高于多元回归光谱重构算法;(2)基于稀疏表达的高光谱重构算法在5种水色传感器上都取得了较好的效果,平均相对误差均在10%以下,均方根误差均在0.005 sr-1以下;(3)相比于原始GOCI多光谱数据,经稀疏表达高光谱重构后的GOCI数据在叶绿素a浓度和总悬浮物浓度估算精度上有不同程度提升.其中对叶绿素a浓度估算而言,平均相对误差从80.6%减少至51.5%,均方根误差从12.175μg·L~(-1)减少至7.125μg·L~(-1);对悬浮物浓度估算而言,平均相对误差从19.1%减少至18.8%,均方根误差从29.048 mg·L~(-1)减少至28.596 mg·L~(-1).     

倏逝波光纤传感器快速检测诺氟沙星

基于间接竞争免疫分析原理,利用倏逝波光纤生物传感平台研发了一种诺氟沙星检测方法,实现了水中诺氟沙星的快速､灵敏检测.研究表明,诺氟沙星检测的优化条件为:抗体浓度为1μg/mL､预反应时间为1min,反应时间为4min.优化条件下,诺氟沙星检测限可达1.89μg/L.包被抗原修饰的光纤探头与荧光标记诺氟沙星抗体具有良好的特异性和稳定性,可重复使用400次以上.自来水､景观水､二沉池出水等水样的加标回收实验结果表明,该传感器具有良好的精密度和准确性,受环境基质的影响较小,能够用于实际水样中诺氟沙星的快速检测.     

贵阳市污水处理厂中典型抗生素的污染水平及生态风险

为了解贵阳市污水处理厂对进水中典型抗生素的去除情况及污水处理厂出水对受纳水体中水生生物的影响,对贵阳市两座污水处理厂进出水及受纳水体中9种典型抗生素进行调查.结果表明,污水处理厂进出水中抗生素均有不同程度检出,进出水中抗生素的浓度范围在ND～835. 60 ng·L~(-1)和ND～286. 60 ng·L~(-1)之间,其中氧氟沙星(OFX)浓度最高,进水分别为835. 60 ng·L~(-1)和539. 00 ng·L~(-1),出水分别为11. 74 ng·L~(-1)和286. 60 ng·L~(-1).污水处理厂对抗生素的去除率为-42. 29%～100%,其中四环素(TC)被完全去除.进出水中抗生素的浓度与国内外其它地区污水处理厂相比处于较低水平.通过对受纳水体中抗生素的检测分析发现,受纳水体中OFX的浓度最高,污水处理厂出水是受纳水体中抗生素的来源之一.通过生态风险评估也发现OFX对受纳水体中水生生物存在高风险(RQ 1).     

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.     

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.     

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.     

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.     

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.     

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.     

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.     

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.     

Removal of heavy metals from sewage sludge by low costing chemical method and recycling in agriculture

ＲｅｍｏｖａｌｏｆｈｅａｖｙｍｅｔａｌｓｆｒｏｍｓｅｗａｇｅｓｌｕｄｇｅｂｙｌｏｗｃｏｓｔｉｎｇｃｈｅｍｉｃａｌｍｅｔｈｏｄａｎｄｒｅｃｙｃｌｉｎｇｉｎａｇｒｉｃｕｌｔｕｒｅＷｕＱｉｔａｎｇ，ＮｙｉｒａｎｄｅｇｅＰａｓｃａｓｉｅ，ＭｏＣｅｈｕｉＦ...