城市生活污水中志贺氏菌ipaH毒力基因的定量PCR检测

基于ipaH毒力基因的实时荧光定量PCR检测,建立适合城市污水中志贺氏菌的定量检测方法。使用从临床分离出的志贺氏菌构建重组质粒作为实时荧光定量PCR的标准品。在ipaH基因模板量2.58×100～2.58×106copy范围内具有良好的线性关系,每100 mL水样中含有2.58×101copy以上的ipaH基因即可被检出。从西安市生活污水分离得到2株野生型志贺氏菌,分析ipaH基因数量和菌体数量的关系,从而确定ipaH基因定量检测志贺氏菌的可行性。该方法灵敏、快速、特异性好,适用于城市生活污水中志贺氏菌的检测。     

城市污水处理工艺对宿主特异性标记物和肠道病原菌的去除效果

采用自制小型反应器模拟实验和实时荧光定量PCR检测方法,针对指示人类、犬类粪便污染的宿主特异性标记物,以及病原性大肠埃希氏菌、沙门氏菌和志贺氏菌等典型的肠道病原菌,分别考察了序批式活性污泥法(SBR)、混凝、过滤工艺对它们的去除效果。研究结果表明,SBR工艺的去除效果优于混凝和过滤工艺,对标记物和病原菌毒力基因的去除率都在93.5%以上。SBR工艺中初期短时间的搅拌即可去除大部分标记物和病原菌。在这3种处理工艺中,宿主特异性标记物与肠道病原菌毒力基因的去除率具有显著相关性,这说明宿主特异性标记物能够在一定程度上反映污水处理过程中肠道病原菌的含量。     

实时荧光定量PCR对A2/O短程硝化系统内氨氧化菌的定量分析

通过控制好氧区低DO浓度以及缩短好氧实际水力停留时间(actual hydraulic retention time,AHRT),在处理低C/N比实际生活污水的A2/O工艺中,成功启动并维持了短程硝化反硝化;系统亚硝酸盐积累率稳定维持在90%左右,氨氮去除率在95%以上。通过提取富集氨氧化菌(ammonia oxidizing bacteria,AOB)的基因组DNA,经两次常规PCR扩增和琼脂糖凝胶电泳,以纯化回收的DNA扩增片段作为实时荧光定量PCR检测AOB数量的DNA标准品,建立了检测AOB数量的实时荧光定量PCR标准曲线。利用实时荧光定量PCR技术比较了A2/O系统在不同运行条件及亚硝酸盐积累率情况下AOB菌群数量。结果表明,随着系统亚硝酸盐积累率的上升,系统内AOB菌群数量也大幅上升。全程硝化和短程硝化时,系统内的AOB菌群数量分别为5.28×109cells/g MLVSS和3.95×1010cells/g MLVSS。此外,亚硝酸盐积累率的下降相对于AOB菌群数量的下降有一定的滞后性。     

水中抑制物对定量PCR与膜过滤法检测E.coli的影响

针对水中病原微生物的污染,选择大肠埃希氏菌E.coli作为病原示踪剂,以E.coli染色体上β-葡萄糖醛酸酶uid A目的基因建立了SYBR Green实时荧光定量PCR的检测方法。该定量PCR方法灵敏度高,检测限可达104CFU/L,线性关系良好,相关系数为R2=0.999。研究表明,定量PCR与菌液浓度呈显著正相关R2=0.935。通过人工投加腐殖酸、COD,研究了水中抑制物对定量PCR和膜过滤MF培养法的影响。结果显示,腐殖酸可使E.coli在培养皿上的菌落变小聚集,且显色不明显,当腐殖酸量为20 mg/L时,浓度为50 CFU/100 m L的E.coli完全受到抑制,没有菌落出现。腐殖酸对PCR扩增的抑制作用明显,当腐殖酸浓度增加到10 mg/L,定量PCR检测结果基因拷贝数减少约1 log,增加到20 mg/L定性PCR检测结果为阴性。     

土霉素生产废水生物处理装置中微生物群落结构特征解析

抗生素对细菌具有强抑制作用,从而会影响废水生物处理系统中的微生物群落结构。在用定量PCR方法对土霉素生产废水处理装置(进水中土霉素浓度为1 662.1±248.6μg/L)中的细菌16S rRNA基因和真菌18S rRNA基因的含量进行比较的基础上,利用16S rRNA基因克隆文库方法对污泥中的细菌群落结构进行了详细解析。定量PCR结果显示,土霉素废水活性污泥中真菌(18S rRNA)/细菌(16S rRNA)基因的拷贝数比例高达1.2,明显较高于非抗生素肌苷废水活性污泥中的比例1.52×10-6,表明真菌对于土霉素废水中有机物的去除可能发挥重要作用。细菌克隆文库分析结果显示,Alpha变形菌和Beta变形菌是主要的优势菌,比例分别为23.7%和22.0%,其次是酸杆菌(17.0%)和拟杆菌(11.9%)。     

水中肠道病毒检测方法的分析评价

采用Hep-2细胞分离培养水中的肠道病毒,根据肠道病毒5’-UTR核酸的保守区,建立了一种细胞培养与实时荧光定量PCR(ICC-RT-q PCR)联合检测感染性肠道病毒的方法。对RT-q PCR、TCID50和ICC-RT-q PCR 3种检测方法进行灵敏度、相关性分析,评价RT-q PCR、ICC-RT-q PCR用于估计水中感染性病毒含量的可行性。结果表明,RT-q PCR方法高估了水中病毒的感染性。肠道病毒低浓度(4.4×10-1~4.4×103TCID50/m L)时TCID50与ICC-RT-q PCR线性关系良好,相关系数R2=0.99。水样经浓缩,ICC-RT-q PCR检测病毒含量为1.0×103~3.2×104copies/L,估计含量为3~30 TCID50/L,与病毒感染性11~29 TCID50/L结果相近,检出率为83.3%。高于TCID50的检测结果(33.3%)。因此,ICC-RT-q PCR方法快速、灵敏,可对水中感染性肠道病毒进行准确的定量分析。     

环境因素对不同工艺人工湿地反硝化功能基因丰度影响

反硝化细菌在人工湿地对N元素去除的过程中发挥着关键作用.为了解湿地类型、基质、植被和水力负荷及物理参数对人工湿地反硝化功能细菌丰度的影响,构建了9个不同工艺组合的人工湿地小试装置.利用实时荧光定量PCR技术检测湿地基质中反硝化功能基因nirS,nirK和nosZ的丰度.结果显示,湿地中nirS,nirK和nosZ丰度(以每ng总DNA中的拷贝数计)分别为4.88×103～2.13×105 copies/ng,1.2×101 ～ 2.14×103 copies/ng和1.98×103～ 2.09 × 105copies/ng,其中nirS显著高于nirK丰度,nirS丰度与湿地出水的pH显著正相关.分析湿地4种设计参数对反硝化功能基因丰度的影响程度,结果显示,对nirS丰度影响程度大小依次为湿地类型＞植被类型＞水力负荷＞基质类型,水平潜流和上行垂直流人工湿地中nirS丰度显著高于下行垂直流人工湿地,无植物人工湿地nirS丰度显著高于有植物人工湿地;对nirK丰度影响程度大小依次为湿地类型＞植被类型＞基质类型＞水力负荷,水平潜流人工湿地nirK丰度显著高于下行垂直流湿地;对nosZ丰度影响程度大小依次为湿地类型＞水力负荷＞植被类型＞基质类型,但影响都不显著.     

生物滴滤塔处理苯乙烯气流的工效和生物膜微群落的分析

采用培养驯化污泥菌种、类球形陶粒和循环液等构建生物滴滤塔.研究评价气体苯乙烯浓度、气体流量、循环液喷淋量对生物滴滤塔工效的影响,并对生物膜微群落中的微种群作了定性定量检测.当进口气体苯乙烯小于1 000 mg/m3、气体流量为200 L/h、循环液流量为10 L/h时,苯乙烯净化效率达90%以上,生化去除量为30 mg/(L·h);单位体积生物膜填料对苯乙烯的最大生化去除量为35 mg/(L·h).湿润生物膜微群落的优势菌种群包括恶臭假单胞菌、梭形芽孢杆菌、罗非氏不动杆菌等5种,恶臭假单胞菌等非芽孢杆菌的最大活菌数为5.5×107 CFU/g,并随生物滴滤塔运行时间延长有减少趋势.     

反相高效液相色谱法测定污水中壬基酚聚氧乙烯醚总量

建立了固相萃取一反相液相色谱一荧光检测法分析污水中壬基酚聚氧乙烯醚总量的方法.固相萃取采用Wa-ters Oasis HLB固相萃取柱(30 μg×60 mg×3 mL),6 mL二氯甲烷/甲醇(90/10,V/V)混合溶剂洗脱,洗脱液用氮吹仪浓缩至1 mL,回收率为99.68%.采用ZORBAX Eclipse XDBC-18(4.6 mm×150 mm,5 μm)反相色谱柱,甲醇/水为流动相,流速1.0 mL/min,以每分钟增加2.5%的梯度陡度进行洗脱,流动相中甲醇比例由70%增加到100%,柱温23℃,进样量15μL.荧光检测器的激发波长230 nm,发射波长313 nm.采用外标法定量.该方法的空白加标和污水样品加标回收率均在90%以上,标准偏差小于10%,定量下限为0.97～1.90 μg/L.利用该方法分别对污水处理厂进水水样、二级处理出水水样、再生水水样进行了测定,测定结果的准确度和精密度满足痕量分析的要求.     

具有降解亚硝酸盐活性的解淀粉芽孢杆菌的分离与安全性分析

从养殖污泥中分离筛选了一株优良的亚硝酸盐降解菌YX01,经ARIS 2X细菌自动鉴定系统以及16SrRNA系统发育分析,该菌株被鉴定为解淀粉芽孢杆菌(Bacillus am yloliquefaciens,GenBank登录号为JX649949),其16S rRNA序列与基因库中 芽孢杆菌属菌株的16S rRNA序列有99％～100％的同源性,而且与解淀粉芽孢杆菌菌株LD5(GenBank登录号:GQ853414)的亲缘关系最近.该亚硝酸盐降解菌YX01在浓度为2.0×107～2.0×109cfu/L时对小球藻(Chlorella sp.)生长有促进作用；在浓度大于2.0×1011cfu/L时对小球藻生长具有显著抑制作用；亚硝酸盐降解菌YX01对小球藻的半数抑制浓度(IC50)大于2.0×1011cfu/L.亚硝酸盐降解菌YX01对大型蚤(Daphnia magna)的48h-半数致死浓度(LC50)大于2.0×1011cfu/L,对草鱼(Ctenopharyngodon idellus)的96h-LC50大于2.0×1011cfu/kg.经分析,亚硝酸盐降解菌YX01为无毒菌株.     

次氯酸钠发生器的研制

介绍了电解法生产次氯酸钠的原理 ,并在原有生产工艺的基础上进行了重新设计和对设备的重新选择、改造 ,得出了各个工艺参数的最佳值 ,生产出高品质的次氯酸钠     

Estimation of uncertainty of sampling for analysis of pesticide residues

Abstract

A computer model was used to take random samples from primary sample populations obtained from field trials to simulate the uncertainty of sampling for residue analysis of plant commodities and soil. The results indicate about 40%, 30% and 20% relative uncertainty when random samples of size 5, 10 and 25 are taken respectively, from a single lot. Therefore the sample size should be the same for establishing and enforcing legal limits.     

Evaluation of the presence of drugs of abuse in tap waters

A total of seventy samples of drinking water were tested for non-controlled and illicit drugs. Of these, 43 were from Spanish cities, 15 from seven other European countries, three from Japan and nine from seven different Latin American countries. The most frequently detected compounds were caffeine, nicotine, cotinine, cocaine and its metabolite benzoylecgonine, methadone and its metabolite EDDP. The mean concentrations of non-controlled drugs were: for caffeine 50 and 19 ng L⁻¹, in Spanish and worldwide drinking water respectively and for nicotine 13 and 19 ng L⁻¹. Illicit drugs were sparsely present and usually at ultratrace level (<1 ng L⁻¹). For example, cocaine has mean values of 0.4 (Spain) and 0.3 ng L⁻¹ (worldwide), whereas for benzoylecgonine, these mean values were 0.4 and 1.8 ng L⁻¹, respectively. Higher concentrations of benzoylecgonine were found in Latin American samples (up to 15 ng L⁻¹). No opiates were identified in any sample but the presence of methadone and EDDP was frequently detected. Total mean values for EDDP were 0.4 ng L⁻¹ (Spain) and 0.3 ng L⁻¹ (worldwide). Very few samples tested positive for amphetamines, in line with the reactivity of chlorine with these compounds. No cannabinoids, LSD, ketamine, fentanyl and PCP were detected.     

Mechanisms of biodegradation of dibenzoate plasticizers

Biodegradation mechanisms were elucidated for three dibenzoate plasticizers: diethylene glycol dibenzoate (D(EG)DB), dipropylene glycol dibenzoate (D(PG)DB), both of which are commercially available, and 1,6-hexanediol dibenzoate, a potential green plasticizer. Degradation studies were done using Rhodococcus rhodochrous in the presence of pure alkanes as a co-substrate. As expected, the first degradation step for all of these systems was the hydrolysis of one ester bond with the release of benzoic acid and a monoester. Subsequent biodegradation of the monobenzoates of diethylene glycol (D(EG)MB) and dipropylene glycol (D(PG)MB) was very slow, leading to significant accumulation of these monoesters. In contrast, 1,6-hexanediol monobenzoate was quickly degraded and characterization of the metabolites indicated that the biodegradation proceeded by way of the oxidation of the alcohol group to generate 6-(benzoyloxy) hexanoic acid followed by β-oxidation steps. This pathway was blocked for D(EG)MB and D(PG)MB by the presence of an ether function.The use of a pure hydrocarbon as a co-substrate resulted in the formation of another class of metabolites; namely the esters of the alcohols formed by the oxidation of the alkanes and the benzoic acid released by hydrolysis of the original diesters. These metabolites were biodegraded without the accumulation of any intermediates.     

混凝试验搅拌器的研制

影响混凝效果的因素众多,混凝沉淀烧杯试验是进行水的混合、絮凝、沉淀工艺研究、设计和生产指导的最有效方法之一,阐述了智能型混凝试验搅拌器的设计原理和技术性能.     

不同泥源对厌氧氨氧化反应器启动的影响

采用2套上流式生物膜反应器,分别接种少量厌氧氨氧化污泥和大量硝化污泥,考察其对厌氧氨氧化反应器启动的影响。污泥接种入反应器后,测得接种厌氧氨氧化污泥的反应器(R1)内MLSS为0.22 g/L,另一个反应器(R2)MLSS为2.7 g/L。与直接接种厌氧氨氧化污泥相比,R1经过72 d的运行才显现出厌氧氨氧化特性。经过114 d的培养,前者氮去除速率由0.23 kg/(m3.d)提升到5.29 kg/(m3.d),总氮去除率大于89%;R2的氮去除速率由0.01 kg/(m3.d)提升到1.1 kg/(m3.d),总氮去除率大于84.6%。说明普通污泥启动需要一个较长的筛选过程,直接接种少量的厌氧氨氧化污泥比接种普通的污泥能够更快启动厌氧氨氧化反应器。     

生物质快速热裂解主要参数对生物油产率的影响

以松木木屑为原料,在自制的小型流化床上,开展了生物质热裂解温度、生物质粒径和进料速率对生物油产率的影响实验研究.结果表明,在热裂解温度分别为450、475、500、525和550℃条件下,当热裂解温度为500℃时,生物油产率最高,平均产率达到53.33%（质量百分比）.反应温度越高,炭产量越低,不可冷凝气体产量越高,气体发热值越高;粒径＜1 mm的生物质其粒径对生物油产率影响不大;生物质进料速率增加时,生物油产率增加.本研究为生物能的利用提供了新的途径.     

An application of the theory of kinematics of mixing to the study of tropospheric dispersion

The most common technique used for numerical simulations of tracer mixing is that of the numerical solution of the advection–diffusion equation with the unresolved fluxes parameterized using the similarity theory. Despite correct predictions of the overall directions of transport, models based on a numerical solution of the advection–diffusion equation lack sufficient accuracy to correctly reproduce the coupling of mixing with small scale processes which are sensitive to the microstructure of the tracer distribution. The objective of this paper is to revisit the basic formalism employed in numerical models used to investigate atmospheric tracers. The main mathematical method proposed here is the theory of kinematics of mixing which could be applied effectively for simulations of atmospheric transport processes. At the beginning of the paper, we introduce simple mathematical transformations in order to demonstrate how complex topological structures are created by mixing processes. These idealistic flow systems are essential to explain transport properties of much more complex three-dimensional geophysical flows. An example of the application of the kinematics of mixing to the analysis of tracer transport on a planetary scale is presented in the following sections. The complex filamentary structures simulated in the numerical experiment are evaluated using some commonly applied statistical measures in order to compare the results with the data published in the literature. The results of the experiment are also analysed with the help of simple conceptual models of fluid filaments. The microstructure of the tracer distribution introduced in the paper is essential to increase our understanding of atmospheric transport and to develop more realistic parameterizations of small-scale mixing. The presented results could also be used to improve calculations of the coupling between microphysical processes and tracer mixing.     

产油微藻培养与回收的关键技术研究进展

寻找廉价而高效的替代原料是实现生物柴油产业化的关键所在.微藻以含油量高、生长周期短、环境适应能力强、生物产量高等优点,有望成为一种极具潜力的生物柴油生产原料.然而,目前尚存在微藻培养低效成本高和微藻回收效率低两大难题.综述了微藻培养与回收过程中的关键技术,并对存在的两大难题及其改进技术进行了详细的探讨.最后,总结并展望了微藻培养、回收技术未来的发展趋势.     

造纸废水混凝处理中SFT助凝替代性研究

中小造纸厂废水处理常用PAC作混凝剂 ,PAM作助凝剂。由于PAM成本很高 ,影响了处理设备的投运率。用超细滑石粉 (SFT)替代PAM助凝 ,与混凝剂PAC配合 ,其混凝处理效果基本相当 ,但是处理成本降低 0 .10元 /m3 。由于SFT属环境无害材料 ,不会给排泥带来二次污染