聚合酶链反应—变性梯度凝胶电泳技术研究梯田式人工湿地微生物群落动态变化

采用聚合酶链反应(PCR)—变性梯度凝胶电泳(DGGE)技术对梯田式人工湿地(TTCW)和复合垂直流式人工湿地(IVCW)生活污水处理系统内的微生物群落结构的动态变化及其对生活污水处理效果的影响进行了对比研究。结果表明:(1)TTCW和IVCW对生活污水均具有较好处理效果,其COD去除率平均分别为76.2%和72.6%、TP去除率平均分别为90.7%和85.6%、NH4+-N去除率平均分别为65.2%和58.8%;TTCW增氧作用明显。(2)2种人工湿地内均存在丰富的微生物群落,并均存在共有的微生物种群;在湿地沿程不同位置均存在一定数量的优势种群。(3)TTCW强化了大气复氧能力,使得湿地内DO较相同深度的IVCW的高。TTCW具有较强的污染物去除能力,特别是具有高效的脱氮除磷能力。这种能力可能主要是TTCW中微生物群落结构的变化所致。     

SBR中SRT对总细菌群落结构的影响研究

为了揭示序批式反应器不同污泥停留时间(SRT)下总细菌群落结构的异同及SRT变化对总细菌群落结构的影响,应用聚合酶链式反应-变性梯度凝胶电泳(PCR-DGGE)进行研究。通过克隆测序发现,不同的SRT条件下生物多样性和种群结构会有所差异,既存在各SRT条件下相同的优势菌群(Escherichia coli和Aeromonas sp.),也存在某些SRT下特有的优势菌群(Uncultured Peptostreptococcaceae),SRT为40 d时检测到以降解硫酸盐获得能源的优势微生物。研究还表明,SRT为40 d时多样性指数取得最大值,各SRT条件下微生物的种群相似性差别较大。     

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.     

空调冷却塔水与空气军团菌的PCR方法快速检测

应用 Enviro Amp PCR-反向杂交法(涉及 5S rRNA基因和 mip基因 )和半巢式 PCR(涉及 16S rRNA基因 )成功检测人工气溶胶化的军团菌 ,这些方法进一步用于检测空调系统冷却塔的水及由其产生的气溶胶中的军团菌 ,从空气样品中检测到军团菌属及嗜肺军团菌 .检测的冷却塔水样品 100%含军团菌属细菌 ,其中 41.6%含嗜肺军团菌 .2种方法相比 ,半巢式 PCR法更经济、方便 .     

降解五氯酚的微好氧颗粒污泥的培养及其微生物种群分析

在微好氧的条件下培养出颗粒污泥并用含五氯酚(PCP)的废水对颗粒污泥进行驯化,研究了培养过程中颗粒污泥的MLSS、SVI、粒径以及对COD和PCP处理能力的变化.颗粒污泥培养成熟后, PCP和COD的去除率分别达到85.3%和86.1%.用扫描电镜观察了颗粒污泥的结构,颗粒污泥内细菌种群丰富,大多数为短杆菌和球菌.同时采用聚合酶链式反应-变性梯度凝胶电泳(PCR-DGGE)技术对培养出的微好氧颗粒污泥以及用PCP驯化后的颗粒污泥中微生物的种群进行了初步分析和对比,用PCP驯化后的微生物种群发生了较明显的动态变化,新增了某些菌群,如不动杆菌(Acinetobacter)、变形杆菌(Proteobacterium)和硫酸盐还原菌等.图6表2参20     

Estrogenic activities of two synthetic pyrethroids and their metabolites

Synthetic pyrethroids(SPs) are among the most common pesticides in current use,and so far,several SPs have been assessed for their potential estrogenicities by various methods.Previous studies have shown that the estrogenicities partly come from their metabolites.Although considerable information is available with respect to the metabolism and environmental degradation of SPs,little is known about the estrogenicities of the metabolites.In this study,permethrin(PM) and β-cypermethrin(CP),as well as their metabolites(3-phenoxybenzoic alcohol(PBCOH),3-phenoxybenzaldehyde(PBCHO) and 3-phenoxybenzoic acid(PBCOOH) were evaluated for their estrogenic activities in the MCF-7 human breast carcinoma cell line.In the MCF-7 cell proliferation assay,PM and CP exhibited significant estrogenic activities at 10-7 mol/L,comparable to 17β-estradiol(E2 ) of 10-9 mol/L,with the relative proliferative effect ratios of 55.4% and 56.3%,respectively.The real-time quantitative polymerase chain reaction(qRT-PCR) results confirmed the estrogenicities of PM and CP with significant alteration of pS2 and ERα mRNA levels observed at 10-6 mol/L.For the three major metabolites,PBCOH and PBCOOH exhibited estrogenic activities in all assays,while no significant estrogenic responses was observed for PBCHO compared to the vehicle control.In particular,PBCOH had even slightly stronger estrogenic activity than its parent compounds,indicating that metabolism may be one of the reasons for the estrogenicities of the SPs.Given the widespread use of SPs,the toxicological effects of parent compounds and their metabolites should be taken into consideration in the risk assessment of SPs.     

Amplification of plasmid DNA bound on soil colloidal particles and clay minerals by the polymerase chain reaction

Polymerase chain reaction(PCR)was used to amplify a 600-base pair(bp)sequence of plasmid pGEX-2T DNA bound on soil colloidal particles from Brown soil(Alfisol)and Red soil(Ultisol),and three different minerals(goethite,kaolinite,montmorillonite). DNA bound on soil colloids,kaolinite,and montmorillonite was not amplified when the complexes were used directly but amplification occurred when the soil colloid or kaolinite-DNA complex was diluted,10- and 20-fold.The montmorillonite-DNA complex required at least 100-fold dilution before amplification could be detected.DNA bound on goethile was amplified irrespective of whether the complex was used directly,or diluted 10- and 20-fold.The amplification of mineral-bound plasmid DNA by PCR is,therefore,markedly influenced by the type and concentration of minerals used.This information is of fundamental importance to soil molecular microbial ecology with particular reference to monitoring the fate of genetically engineered microorganisms and their recombinant DNA in soil environments.     

通讯

纳米银通过结合RNA聚合酶抑制红系前体细胞RNA的转录由于其广谱的抗菌活性,纳米银在日常生活和医药卫生中得到广泛的应用.纳米银的大量生产和使用将导致其在生产、运输、使用和排放等环节释放入环境,其潜在的环境与健康风险已日益受到关注.研究发现,纳米银可以引发呼吸系统(肺)、消化系统(肝脏)、神经系统(神经元)和生殖系统的组织损伤.然而,目前对纳米银毒性效应的分子作用机制尚待深入研究.     

北江河水中抗生素抗性基因污染初步研究（Preliminary Studies on the Pollution Levels of Antibiotic Resistance Genes in the Water of Beijiang River, South China）

环境中细菌的耐药性,尤其是对抗生素的耐药性已成为影响生态环境的重要因素.论文采用抗生素抗性平板法调查了北江河水中四环素、红霉素及磺胺类这3类抗生素耐药性细菌的存在,采用定性PCR及荧光定量PCR方法分别研究了该水域sul1和sul2这2种磺胺类抗生素抗性基因(ARGs)的存在和含量水平.结果表明,所采集北江水域的9个样品中有5个对四环素有耐药性,7个对红霉素有耐药性,8个对磺胺二甲嘧啶有耐药性;定性PCR实验并经基因测序结果证实,5个样品含有sul1,4个样品含有sul2.进一步的PCR定量分析结果显示,7个样品中均检出sul1和sul2磺胺抗性基因,它们与内对照基因16S-rRNA表达量比值分别在10-2.56~10-0.52及10-3.25~10-1.24范围内,该结果显著高于美国科罗拉多州北部河流的研究结果.此外,数据分析也发现,sul1和sul2磺胺抗性基因的含量水平与该区域水中磺胺含量分布具有一定的相关性,表明外源性抗生素对河流的污染是诱导抗性基因的重要因素.     

动物源性食品中抗生素类污染物生物检测技术研究进展

抗生素类污染物在动物源性食品中的残留,因其对人类健康有着潜在危害而受到了广泛的关注,因此迫切需要建立快速、灵敏、高效、便捷的检测方法以便对其及时管理和防治.生物检测技术在实现对抗生素高灵敏度、高特异性、低成本、快速的检测方面具有很强的优势,主要包括微生物检测法、免疫分析法和生物传感器法等.本文基于免疫抗体和核酸适配体两...