采用发光菌和统计法评估地下水水质 ——以“珠江三角洲”滨海小流域为例

以珠江三角洲滨海小流域地下水为研究对象,采用费氏弧菌(Vibrio ficheri)和化学指标主成分分析法综合评价不同人类活动背景下的地下水(8个饮用水井和6个观测井)以及海水水质.结果表明:15个水样中的阳离子以K+和Na+为主,占阳离子总量的平均百分比为71.55%±14.67%.阴离子的浓度差异较大,饮用水井(T1~T8)的阴离子以HCO3-为主,观测井(D1~D6)的阴离子以Cl-为主,平均百分比分别为53.84%±18.21%和66.37%±22.16%.除T3、T4和D3无毒外,其余水样对发光菌具有一定的抑制效应,其中D2和D4对发光菌的抑制率大于30%,为中毒性,海水水样则对发光菌产生了“刺激发光”效应,30min的发光抑制率为-12.45%;通过水样化学指标的主成分分析,计算井水的综合得分,排泄区D2~D4 (8~15m潜水含水层)得分较高,D5 (承压含水层)、D6 (基岩裂隙含水层)以及D1(5m潜水含水层)得分较低,唐家镇除T1地下水埋深最深(17.92m),得分最高,其余地下水(T2~T8)位于地表以下10m内的含水层,得分较低;发光抑制率与第一主成分具有良好的负相关性(P<0.05),经拟合方程预测的毒性结果准确率为75%,“假阴性”水样个数为0.     

二级处理出水的UV-TiO2消毒及大肠杆菌和粪肠球菌光复活试验

对污水处理厂二级处理出水进行UV和UV-TiO2光催化消毒,研究了复活光强度(0~41μW·cm-2)对不同方法消毒后水中大肠杆菌和粪肠球菌复活的影响.结果表明,UV-TiO2消毒效率远高于单纯的UV消毒.大肠杆菌经UV-TiO2消毒后,只有在较强的复活光照射下才会产生一定的光复活,而单纯UV消毒后即使在较低的复活光强下也会出现较强的光复活.UVTiO2消毒对粪肠球菌的光复活现象抑制明显,在小于21μW·cm-2的低复活光照射下粪肠球菌经72 h照射后几乎不发生光复活现象,只有在较强的复活光照射下才会产生一定的光复活.     

UV和H2 O2联合消毒灭活饮用水中大肠杆菌研究

以大肠杆菌为研究对象,对单独紫外线(UV)、H2O2及两者不同组合方式的灭活效果进行了分析.结果表明,单独H2O2对大肠杆菌基本无灭活效果,加入浓度为20 mg·L-1的H2O2接触时间为30 min时,灭活率仅为0.02个对数级;单独UV工艺可在一定程度上灭活大肠杆菌,紫外线剂量为10 mJ·cm-2时,灭活率可达到4.51个对数级.紫外线和H2O2联合消毒在一定程度上提高了消毒效果,且不同的组合方式对灭活效果有很大的影响.先UV后H2O2消毒效果优于先H2O2后UV,当紫外线剂量为5 mJ·cm-2时,加入H2O2分别反应2.5、5、10、20 min后,两者的灭活率分别较单独UV消毒增加0.09、0.35、0.38、0.68和0.01、0.07、0.14、0.53个对数级.而UV与H2O2同时消毒效果优于UV与H2O2顺序消毒,当紫外线剂量为5 mJ·cm-2时,加入H2O2反应20 min后,灭活率较UV-H2O2和H2O2-UV工艺分别增加了0.43和0.58个对数级.随着紫外线强度的增加,大肠杆菌的灭活效果不断提高.     

Quantification of viable bacteria in wastewater treatment plants by using propidium monoazide combined with quantitative PCR (PMA-qPCR)

The detection of viable bacteria in wastewater treatment plants(WWTPs) is very important for public health, as WWTPs are a medium with a high potential for waterborne disease transmission. The aim of this study was to use propidium monoazide(PMA) combined with the quantitative polymerase chain reaction(PMA-qPCR) to selectively detect and quantify viable bacteria cells in full-scale WWTPs in China. PMA was added to the concentrated WWTP samples at a final concentration of 100 μmol/L and the samples were incubated in the dark for 5 min, and then lighted for 4 min prior to DNA extraction and qPCR with specific primers for Escherichia coli and Enterococci, respectively. The results showed that PMA treatment removed more than 99% of DNA from non-viable cells in all the WWTP samples, while matrices in sludge samples markedly reduced the effectiveness of PMA treatment. Compared to qPCR, PMA-qPCR results were similar and highly linearly correlated to those obtained by culture assay, indicating that DNA from non-viable cells present in WWTP samples can be eliminated by PMA treatment, and that PMA-qPCR is a reliable method for detection of viable bacteria in environmental samples. This study demonstrated that PMA-qPCR is a rapid and selective detection method for viable bacteria in WWTP samples, and that WWTPs have an obvious function in removing both viable and non-viable bacteria. The results proved that PMA-qPCR is a promising detection method that has a high potential for application as a complementary method to the standard culture-based method in the future.     

Comparative cytotoxicity of nanoparticles and ions to Escherichia coli in binary mixtures

The objective of this study was to understand toxicity of mixture of nanoparticles (NPs) (ZnO and TiO₂) and their ions to Escherichia coli. Results indicated the decrease in percentage growth of E. coli with the increase in concentration of NPs both in single and mixture setups. Even a small concentration of 1 mg/L was observed to be significantly toxic to E. coli in binary mixture setup (exposure concentration: 1 mg/L ZnO and 1 mg/L TiO₂; 21.15% decrease in plate count concentration with respect to control). Exposure of E. coli to mixture of NPs at 1000 mg/L (i.e., 1000 mg/L ZnO and 1000 mg/L TiO₂) resulted in 99.63% decrease in plate count concentration with respect to control. Toxic effects of ions to E. coli were found to be lesser than their corresponding NPs. The percentage growth reduction was found to be 36% for binary mixture of zinc and titanium ions at the highest concentration (i.e., 803.0 mg/L Zn and 593.3 mg/L Ti where ion concentrations are equal to the Zn ions present in 1000 mg/L ZnO NP solution and Ti^+ 4 ions present in 1000 mg/L TiO₂ NP solution). Nature of mixture toxicity of the two NPs to E. coli was found to be antagonistic. The alkaline phosphatase (Alp) assay indicated that the maximum damage was observed when E. coli was exposed to 1000 mg/L of mixture of NPs. This study tries to fill the knowledge gap on information of toxicity of mixture of NPs to bacteria which has not been reported earlier.     

电化学消毒法对水中大肠杆菌的灭活特性

本研究通过电化学消毒小试装置,系统考察了电极板材料、极板间距、p H、水温、Na Cl浓度等因素对水中大肠杆菌CGMCC 1.3373灭活效果的影响,并进一步考察了电化学消毒对抗生素抗性大肠杆菌CGMCC 1.1595的灭活效果及其抗性变化.结果发现,在相同的电化学消毒条件下,阳极采用钌铱涂层的钛电极板的灭活效果显著优于铅涂层和铱钽涂层电极板,并且极板间距为50 mm时灭活率最高.进一步研究表明,电化学消毒法对大肠杆菌的灭活率随水的p H升高而降低,随水温和Na Cl浓度升高而升高.在相同消毒条件下,电化学法对四环素抗性大肠杆菌的灭活率显著低于普通大肠杆菌.消毒后存活的抗性菌对四环素、青霉素、氯霉素的抗性,随消毒时间延长呈现先下降后上升的规律.     

微生物杀灭效果试验中的游离氯转化研究

在实验室研究游离氯灭活微生物的试验中发现,由于试验体系中有机氮的存在,投加游离氯消毒后,游离氯会迅速与之反应转化为无消毒作用的有机氯胺.在对大肠杆菌、铁细菌、铜绿假单胞菌的研究中均发现同样的问题,投加2 mg·L^-1的游离氯消毒剂对初始菌浓度为10⁸ CFU·mL^-1的大肠杆菌进行灭活试验,反应5　min后游离氯就已经为0,而一氯胺和二氯胺的量分别为0.92　mg·L^-1和0.4 mg·L^-1.为了降低有机氮的干扰,采用多次离心纯化菌液、膜过滤纯化菌液和增加受试菌液的稀释倍数的方法进行测试,发现当菌液浓度降至10⁵CFU·mL^-1以下时,游离氯转氯胺的比例可以明显降低;而2种纯化过程并不能分离菌液中的有机氮,进而改善试验中出现的游离氯转氯胺的现象.该结论也说明参加反应的有机氮应来源于细菌自身的细胞物质,有机氮的干扰在消毒的相关研究中需要引起关注.     

连续流光催化消毒器灭活E.coli及病毒的性能

紫外消毒是污水处理常用技术.紫外线仅能破坏微生物的遗传物质,阻断其繁殖,并不能彻底杀死微生物,一旦停止紫外光照射,微生物可在修复酶的作用下恢复活性.光催化过程能产生强氧化性·OH,破坏细胞壁和细胞膜,彻底杀死微生物.为考察光催化技术消毒效率、抑制复活性能及应用潜力,在市售紫外消毒器中安装了光催化组件,搭建了连续流动式光催化消毒器.以大肠杆菌(E.coli)和噬菌体为目标物评价其消毒能力.紫外剂量50 m J·cm-2条件下,初始浓度3.40×106CFU·m L-1的E.coli原水流经紫外消毒器后被去除4.12 log10的E.coli,而流经光催化消毒器后去除4.79 log10.同时光催化处理后E.coli的光复活率仅为紫外消毒后的17%.连续运行40 h的灭菌能力稳定.光催化设备对噬菌体F2和噬菌体MS2的去除率分别为6.37 log10和6.00 log10,表明该设备也具有病毒去除能力.     

南方某城市地表水体中粪源性污染指示微生物的分布特征研究

随着城市居民亲水娱乐活动日益增加,研究城市地表水体的粪源性微生物污染特征,开展人体健康风险评价,并以此为基础开展有效的水质监测和风险管理,对于人口密集的城市区域尤为重要.以我国南方某城市的5个典型地表水体为案例,监测总大肠菌（TC）、粪大肠菌（FC）、大肠埃希氏菌（EC）和肠球菌（ENT）这4种粪源性污染指示微生物以及常规理化水质指标,研究指示微生物在城市地表水体中的浓度水平、时间和空间分布特征以及不同水质指标之间的相关性.结果表明,案例水体中TC和FC通常在10^3～10^7个.L^-1,EC和ENT通常分别低于200个·L^-1和105个·L^-1;4种指示微生物的浓度分布呈现季节变化特征,并且能够指示水体受到周边环境的污染状况;4种指示微生物指标之间以及微生物指标与常规理化水质指标（如溶解氧等）之间具有一定的相关性.     

生物传感细胞ADP1_pWHlux在水环境急性毒性检测中的应用

针对天然发光菌和以模式微生物为宿主构建的生物传感细胞在急性毒性检测应用中对测试条件要求苛刻等适用性问题,本研究将1株基因工程构建的生物传感细胞不动杆菌ADP1_p WHlux应用于急性毒性检测,建立检测方法,考察其灵敏度及检测范围.结果表明,ADP1_p WHlux发光受急性毒物的抑制,毒物剂量与发光抑制存在剂量效应关系.在4 mg·L-1Hg Cl2诱导下仅5 min可作出响应,暴露30~60 min后可以给出较为准确的结果.对Hg Cl2的检出限可达0.04 mg·L-1.对我国生活饮用水卫生标准的指标中Be2+、Ba2+、Cu2+、Ni2+检出效果明显,对Be2+、Ba2+、Cu2+的检测范围均在0.025~250 mg·L-1,对Ni2+的检测范围在0.002 5~250 mg·L-1,对Pb2+、Br O-3、Cl O-2的检出限均在0.002 5 mg·L-1,对Cl O-3检出限为0.025mg·L-1.采用ADP1_p WHlux生物传感细胞检测方法对北京市清河水环境急性毒性进行评价,表明ADP1_p WHlux生物传感细胞检测方法可用于污染水样检测.     

A GFP-based bacterial biosensor with chromosomally integrated sensing cassette for quantitative detection of Hg(II) in environment

A mercury biosensor was constructed by integrating biosensor genetic elements into E. coli JM109 chromosome in a single copy number, using the attP/attB recombination mechanism of λphage. The genetic elements used include a regulatory protein gene (merR) along with operator/promoter (O/P) derived from the mercury resistance operon from pDU1358 plasmid of Serratia marcescens. The expression of reporter gene gfp is also controlled by merR/O/P. Integration of the construct into the chromosome was done to increase the stability and precision of the biosensor. This biosensor could detect Hg(II) ions in the concentration range of 100-1700 nmol/L, and manifest the result as the expression of GFP. The GFP expression was significantly different (P ≤ 0.05) for each concentration of inducing Hg(II) ions in the detection range, which reduces the chances of misinterpretation of results. A model using regression method was also derived for the quantification of the concentration of Hg(II) in water samples.     

双级虚拟撞击采样器应用于固定污染源PM10和PM2.5排放测量

为贯彻落实《大气污染防治行动计划》,环境保护部指导各地开展大气污染源排放清单编制工作,其中包括固定源PM_(10)和PM_(2.5)的排放清单.但目前国内尚无固定源PM_(10)和PM_(2.5)标准采样方法.本研究提出了适合我国固定源PM_(10)和PM_(2.5)测量的双级虚拟撞击采样方法,开发了相应的分级采样系统,并用该方法对各类固定源进行了现场测试.测试结果表明,所测试的煤粉炉电厂的烟囱入口PM_(2.5)质量浓度为(0.93±0.03)mg·m~(-3),PM_(10)质量浓度为(1.13±0.11)mg·m~(-3).所测试的垃圾焚烧电厂的烟囱排放口PM_(2.5)质量浓度为(3.3±0.65)mg·m~(-3),PM_(10)质量浓度为(6.9±0.86)mg·m~(-3).所测试的大型循环流化床发电厂的烟囱排放口PM_(2.5)质量浓度为(0.59±0.04)mg·m~(-3),PM_(10)质量浓度为(1.12±0.16)mg·m~(-3).所测试的钢铁转炉的烟囱排放口PM_(2.5)质量浓度为(0.15±0.04)mg·m~(-3),PM_(10)质量浓度为(0.43±0.15)mg·m~(-3).     

固定化混合菌Z1对制革废水Cr3+和OPnEO的同步去除及其响应面法优化

以微紫青霉菌B5和醋酸钙不动杆H1按照1∶1配置成混合菌Z1,并分别以聚乙二醇、改性秸秆、海藻酸钠对混合菌Z1进行包埋,探究其对Cr~(3+)和OPnEO的同步去除处理情况,结果得到OPnEO的去除率分别为53.42%、46.29%、55.14%;而Cr~(3+)的去除率分别达到58.41%、54.24%、60.95%,确定海藻酸钠为Z1的最佳包埋材料.以Cr~(3+)去除率为响应值,利用Plackett-Burman实验确定Cr~(3+)的初始浓度、pH值、温度为影响混合菌Z1去除Cr~(3+)的3个主要因素;运用最陡爬坡实验设计,结合Box-Behnken实验设计及响应面法分析,确定海藻酸钠固定化混合菌Z1在初始pH 7.64、Cr~(3+)初始浓度856.34 mg·L~(-1)、温度30.39℃、OPnEO初始浓度950 mg·L~(-1)、外加2 m L胰蛋白胨条件下培养7 d,Cr~(3+)的去除率达到77.69%,比单因素实验条件下Cr~(3+)的去除率提高7%.