FDA水解酶分析法表征近海泥滩微生物活性

针对近海潮间带环境高盐、有机质组成复杂和微生物活性低的特点,根据微生物酶解荧光素双醋酸酯(FDA)产生荧光素的原理,建立了适于泥滩类潮间带沉积环境中微生物活性检测的荧光光度法.从样品浸提液、反应产物检测仪器、预处理方法、实验条件等不同方面进行优化,确定泥滩类潮间带沉积环境中微生物活性的最优检测方法和条件如下:沉积物湿样以灭菌陈海水为介质,加入吐温-80分散剂,充分振荡后静置,使较大颗粒自然沉降;取上层悬浊液,经无菌滤膜(1.2μm,多次煮沸灭菌)过滤,得到待测菌液;取待测菌液加入适量FDA溶液,在25~30℃下避光反应180 min,以丙酮为终止剂终止反应,25 min内以分子荧光光度计(激发波长488 nm,发射波长530 nm)测定反应产物的荧光强度,该方法的检测范围(以干重计)是3.0×103~1.1×105个·g-1.微生物活性以单位质量样品的荧光素含量表示(μg·g-1,干重).     

水中大肠菌群微滴数字PCR定量检测方法的建立

以大肠菌群的lacZ基因为靶基因,建立一种快速、稳定、灵敏、特异的微滴数字聚合酶链式反应(Droplet Digital Polymerase Chain Reaction,ddPCR)定量检测方法。对ddPCR反应体系中试剂浓度、退火温度等条件优化筛选的同时,考察了方法的线性范围、精密度和定量限。最终确定反应体系中的最佳引物和探针浓度分别为0.2和0.5μmol/L,最佳退火温度为56℃。大肠菌群lacZ基因组DNA浓度范围为3.95～7.80×10~4拷贝(copies)/20μL ddPCR反应液时,方法的线性关系良好(R~2=0.999)。文章所建立的方法可检出每微升单个拷贝数的大肠菌群lacZ基因组DNA,且具有快速测定、灵敏度高、特异性强、重复性良好等特点,为建立水污染早期应急机制提供了重要的参考价值。     

高温水萃取在食品安全快速检测中的应用研究

采用高温水萃取方法对食品中农药残留、甲醛、亚硝酸盐、抗生素等成分进行快速萃取,结合分光光度法、液相色谱等分析方法进行了上述成分的快速检测研究。结果表明,蔬菜中甲胺磷的萃取条件为pH=8.0的磷酸缓冲溶液、萃取温度120℃、萃取时间2～3min;腐竹中甲醛的萃取条件为0.1mLH3PO4溶液/10mL、萃取温度120℃、萃取时间5min;肉制品中亚硝酸盐的萃取条件为5mL硼砂饱和溶液/10mL、萃取温度120℃、萃取时间5min;饲料中四环素的萃取条件为pH=2.0的盐酸溶液、萃取温度100℃、萃取时间4min。与现有的蒸馏、振荡萃取、超声萃取等预处理方法比较,高温水萃取法具有萃取效率高、精密度好、快速、简便和不使用有机溶剂的优点,可以满足实际样品快速检测的需要。     

环介导等温扩增(LAMP)技术检测水源病原菌

微生物指标是水质安全的重要指标之一,但是由于技术原因,大部分病原微生物并未被纳入水质监测中。目前各类水质标准中只有病原指示微生物的指标,然而这些指标与病原微生物的相关性并不理想。因此,开发快速简便的病原微生物检测技术是改进水质生物检测和监测的关键。环介导等温扩增技术是一种快速检测DNA且不依赖昂贵检测设备的PCR技术。尝试利用该技术检测水中的8种常见指示菌和病原菌:总大肠杆菌、沙门氏菌、鲍氏志贺氏菌、大肠埃希氏菌O157∶H7、小肠耶尔森氏菌、嗜肺军团菌、绿脓杆菌和结核杆菌,获得了针对其中5种细菌的毒力基因的特异性引物和最优等温PCR条件。环介导等温扩增技术的灵敏度可在45min内检测出100个基因拷贝,并能在原污水中对病原菌进行定性检测。     

金/钴氢氧化物膜修饰玻碳电极同时测定邻苯二酚和对苯二酚

文章将纳米金与钴氢氧化物膜的催化作用有效结合,制备了GNPs/CoOOH复合修饰电极,该修饰电极在碱性条件下对邻苯二酚和对苯二酚具有较强的电催化活性。考察了支持电解质酸度及纳米金沉积时间对邻苯二酚和对苯二酚电化学响应的影响,选取0.1 mol/L PBS(pH 10.0)作为支持电解质,纳米金的最佳沉积时间为4 min。在优化的实验条件下,利用差示脉冲伏安法(DPV)对邻苯二酚和对苯二酚进行选择性检测:当两者浓度同时改变时,对苯二酚和邻苯二酚的氧化峰电流与其浓度分别在7~100μmol/L和6~100μmol/L范围内呈良好的线性关系,对应的检出限分别为0.9、0.8μmol/L(S/N=3)。该复合修饰电极具有较好的重现性、稳定性及较强的抗干扰能力。     

纳米Fe、Si体系对3,3′,4,4′-四氯联苯的脱氯降解

研究了纳米Fe、Si体系降解3,3′,4,4′-四氯联苯(PCB77)的动力学差异.结果表明,纳米Fe0、纳米Fe3O4和纳米Si0对PCB77均有降解作用,该降解为还原脱氯反应.降解过程符合准一级反应动力学,反应速率常数Kobs分别为0.0177,0.0038,0.0045h-1.PCB77初始浓度为5mg/L,纳米材料投加量为5g/L,溶液pH4.5条件下,纳米Fe0体系对PCB77降解效果最为显著,64h时PCB77残留率仅为19.83%,氯离子浓度为50.3μmol/L,反应体系pH值从4.5升至5.26.纳米双元体系Fe0和Si0、Fe3O4和Si0对PCB77降解过程也符合准一级反应动力学,反应速率常数Kobs分别为0.0114,0.004h-1,其中纳米Fe0和Si0体系降解效果优于纳米Fe3O4和Si0体系.PCB77残留率分别为34.91%和66.62%,氯离子浓度分别为40.07,20.47μmol/L,反应体系pH值变化不明显.随着溶液初始pH值增加,纳米Fe0、纳米Fe3O4降解PCB77效果明显降低,但溶液pH值升高有利于纳米Si0对PCB77的降解.两组纳米双元体系对PCB77的降...     

九龙江下游水源水中新发病原微生物和抗生素抗性基因的定量PCR检测

新发水传病原微生物和抗生素抗性基因对于饮用水的生物安全性有重要意义.以7种典型的水传病原微生物和4种抗生素抗性基因作为检测指标,对闽西南地区的重要水源九龙江下游水源水进行了检测.从九龙江下游4个采样点和附近某水厂进出水采集水样,用0.22μm的滤膜过滤后,进行DNA提取及荧光定量PCR检测.结果表明,九龙江下游水源水中Salmonella enterica(Salmonella spp.)、Legionella pneumophila(L.pneumophila)和Pseudomonas aeruginosa(P.aeruginosa)等个别指标可达到103、104、105copies.mL-1的水平.有机物浓度可能对病原微生物和抗生素抗性基因的水平有较大影响.水厂常规工艺对于大多数病原微生物和抗生素抗性基因可以实现有效去除,但对Salmonella spp.的去除效果不理想.因此为保证饮用水安全,需要在清理污染源、提高处理工艺水平以及末端控制方面加强管理.     

高效液相色谱法测定工业废水中邻苯二胺

通过实验建立了工业废水中邻苯二胺的高效液相色谱分析方法.采用Kromasii-C18(150mm×4.6mm,5μm)色谱柱,流速1 mL/min,柱温30 ℃,进样体积10μL,流动相为V(甲醇):V(0.01 mol/L醋酸铵溶液)=35:65,测定波长290 nm.结果线性范围为:1-100 mg/L,平均加标回收率为101.2%,相对标准偏差为0.75%.     

油田污水中可溶性硫化物快速检测方法

采用硫离子选择电极法快速检测油田水中硫化物,优化了操作条件,在溶液pH值大于11.5和中速搅拌（800 r/min左右）条件下,单个样品分析时间少于15 min.空气中的氧对测定结果干扰明显,通过取样时加入硫化物稳定剂方法可消除空气中氧的影响.方法的精密度小于4.0％,与碘量法相比,相对偏差小于15％,方法检测下限为0.35 mg/L.     

多壁碳纳米管修饰电极对药品中扑热息痛的测定

该文首先对比了扑热息痛(PCT)在多壁碳纳米管、金纳米颗粒以及金纳米颗粒与碳纳米管混合掺杂等不同纳米材料修饰氧化铟锡(ITO)电极表面的电化学反应行为,确定多壁碳纳米管对PCT的催化作用后,对溶液p H、扫描速度等测定条件进行优化。在最佳响应条件下,运用方波伏安法(SWV)在5.0×10-9~100.0×10-9mol/L浓度范围内,建立PCT浓度与响应电流的线性方程。并将该修饰电极成功用于5种治疗流感药品中扑热息痛浓度的快速测定。     

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.     

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.     

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.     

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

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Isolation and preliminary characterization of a 3-chlorobenzoate degrading bacteria

A study was conducted to compare the diversity of 2-, 3-, and 4-chlorobenzoate degraders in two pristine soils and one contaminated sewage sludge. These samples contained strikingly different populations of mono-chlorobenzoate degraders. Although fewer cultures were isolated in the uncontaminated soils than contaminated one, the ability of microbial populations to mineralize chlorobenzoate was widespread. The 3- and 4-chlorobenzoate degraders were more diverse than the 2-chlorobenzoate degraders. One of the strains isolated from the sewage sludge was obtained. Based on its phenotype, chemotaxonomic properties and 16S rRNA gene, the organism S-7 was classified as Rhodococcus erythropolis. The strain can grow at temperature from 4 to 37℃. It can utilize several （halo）aromatic compounds. Moreover, strain S-7 can grow and use 3-chlorobenzoate as sole carbon source in a temperatures range of 10-30℃ with stoichiometric release of chloride ions. The psychrotolerant ability was significant for bioremediation in low temperature regions. Catechol and chlorocatechol 1,2-dioxygenase activities were present in cell free extracts of the strain, but no （chloro）catechol 2,3- dioxygenase activities was detected. Spectral conversion assays with extracts from R. erythropolis S-7 showed accumulation of a compound with a similar UV spectrum as chloro-cis,cis-muconate from 3-chlorobenzoate. On the basis of these results, we proposed that S-7 degraded 3-chlorobenzoate through the modified ortho-cleave pathway.     

Single and joint effects of pesticides and mercury on soil urease

The influence of two pesticides including chlorimuron-ethyl and furadan and mercury （Hg） on urease activity in 4 soils （meadow burozem and phaeozem） was investigated. The soils were exposed to various concentrations of the two pesticides and Hg individually and simultaneously. Results showed that there was a close relationship between urease activity and organic matter content in soil. Chlorimuron-ethyl and furadan could both activate urease in the 4 soils. The maximum increment of urease activity by chlorimuronethyl was up to 14%-18%. There was almost an equal increase （up to 13%-21%） in the urease activity by furadan. On the contrary, Hg markedly inhibited soil urease activity. A logarithmic equation was used to describe the relationship （P〈0.05） between the concentration of Hg and the activity of soil urease in the 4 tested soils. Semi-effect dose （ED50） values by the stress of Hg based on the inhibition of soil urease in the 4 soils were 88, 5.5, 24 and 20 mg/kg, respectively, according to the calculation of the corresponding equations. The interactive effect of chlorimuron-ethyl or furadan with metal Hg on soil urease was mainly synergic at the highest tested concentrations.     

Organochlorine pesticides in soils under different land usage in the Taihu Lake region, China

A field study was conducted in the Taihu Lake region, China in 2004 to reveal the organochlorine pesticide concentrations in soils after the ban of these substances in the year 1983. Thirteen organochlorine pesticides (OCPs) were analyzed in soils from paddy field, tree land and fallow land. Total organochlorine pesticide residues were higher in agricultural soils than in uncultivated fallow land soils. Among all the pesticides, ΣDDX (DDD, DDE and DDT) had the highest concentration for all the soil samples, ranging from 3.10 ng/g to 166.55 ng/g with a mean value of 57.04 ng/g and followed by ΣHCH, ranging from 0.73 ng/g to 60.97 ng/g with a mean value of 24.06 ng/g. Dieldrin, endrin, HCB and α-endosulfan were also found in soils with less than 15 ng/g. Ratios of p,p'-(DDD DDE)/DDT in soils under three land usages were: paddy field ＞ tree land ＞ fallow land, indicating that land usage inlfuenced the degradation of DDT in soils. Ratios of p,p'-(DDD DDE)/DDT ＞1, showing aged residues of DDTs in soils of the Taihu Lake region. The results were discussed with data from a former study that showed very low actual concentrations of HCH and DDT in soils in the Taihu Lake region, but according to the chemical half-lives and their concentrations in soils in 1980s, the concentration of DDT in soils seemed to be underestimated. In any case our data show that the ban on the use of HCH and DDT resulted in a tremendous reduction of these pesticide residues in soils, but there are still high amounts of pesticide residues in soils, which need more remediation processes.     

Alterations of organ histopathology and metallolhionein mRNA expression in silver barb, Puntius gonionotus during subchronic cadmium exposure

Common silver barb,Puntius gonionotus,exposed to the nominal concentration of 0.06 mg/L Cd for 60 d,were assessed for histopathological alterations(gills,liver and kidney),metal accumulation,and metallothionein(MT)mRNA expression.Fish exhibited pathological symptoms such as hypertrophy and hyperplasia of primary and secondary gill lamellae,vacuolization in hepatocytes,and prominent tubular and glomerular damage in the kidney.In addition,kidney accumulated the highest content of cadmium,more than gills and liver.Expression of MT mRNA was increased in both liver and kidney of treated fish.Hepatic MT levels remained high after fish were removed to Cd-free water.In contrast,MT expression in kidney was peaked after 28 d of treatment and drastically dropped when fish were removed to Cd-free water.The high concentrations of Cd in hepatic tissues indicated an accumulation site or permanent damage on this tissue.