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1.
PCR-DGGE技术解析生物制氢反应器微生物多样性   总被引:18,自引:9,他引:18  
为了揭示发酵法生物制氢反应器厌氧活性污泥的微生物种群多样性 ,从运行不同时期取厌氧活性污泥 ,通过细胞裂解直接提取活性污泥的基因组DNA .以细菌 16SrRNA基因通用引物F338GC/R5 34进行V3高变异区域PCR扩增 ,长约 200bp的PCR产物经变性梯度凝胶电泳 (DGGE)分离后 ,获得微生物群落的特征DNA指纹图谱 .研究表明 ,不同时期的厌氧活性污泥中存在共同种属和各自的特异种属 ,群落结构和优势种群数量具有时序动态性 ,微生物多样性呈现出协同变化的特征 .微生物多样性由强化到减弱 ,群落结构之间的相似性逐渐升高 ,演替速度由快速到缓慢 .优势种群经历了动态演替过程 ,最终形成特定种群构成的顶级群落 .  相似文献   

2.
采用聚合酶链式-变性梯度凝胶电泳(PCR-DGGE)技术,研究了膜生物反应器(MBR)和传统活性污泥工艺(CAS)反应器中微生物在贫营养条件下的总细菌群落结构.结果表明,在培养过程中,污泥的微生物种群经历了一个比较明显的变化过程,且以CAS污泥微生物种群的变化更为明显,演替过程中既有原始优势种群的消亡,又有新的优势种群...  相似文献   

3.
MBR中微生物群落结构的演变与分析   总被引:17,自引:3,他引:14       下载免费PDF全文
张斌  孙宝盛  季民  赵祖国 《环境科学学报》2008,28(11):2192-2199
为了揭示膜.生物反应器中微生物群落结构多样性的演变过程,通过细胞裂解法直接提取不同时期污泥中的基因组DNA,利用基于16SrDNA的PCR-DGGE技术获得了微生物群落的DNA特征指纹图谱,并对条带进行了统计分析和切胶测序,使用序列数据进行了同源性分析并建立了系统发育树.DGGE分析表明,在反应器运行前17d内污泥中微生物群落结构变化很大,与接种污泥的相似性系数下降到了29.2%,从而说明MBR中处理工艺和进水水质的改变导致微生物群落结构多样性降低.在试验过程中,Pscudomonas和Aeromonas hydrophila等种群一直保持着较为稳定的优势地位,也有原始种群如Bacillus sp.的消亡和以Enterococcus faecalis、Comamonas sp.、Fusobacterium sp.等为代表的次级种群的强化和演变.UPGMA聚类分析将DGGE图谱区分为3大类群并对应于各自的运行时期.测序结果表明,MBR中微生物菌群间进化距离较大,其中Proteobaeteria纲和Bacillus属细菌较多.在反应器运行后期演变为优势地位的菌群(如Comamonas sp.)加剧了膜污染物的产生和积累.  相似文献   

4.
处理不同废水MBR系统中微生物群落结构的比较   总被引:10,自引:4,他引:6       下载免费PDF全文
张斌  孙宝盛  刘慧娜  刘宪华  季民 《环境科学》2008,29(10):2944-2949
为了研究膜生物反应器中微生物群落结构与系统处理效能的关系.为工艺改进提供依据,从4种处理不同水质的MBR污泥中提取细菌总基因组DNA,采用PCR-DGGE和克隆测序技术对系统中的微生物群落结构进行了解析,根据序列数据进行同源性分析并建立了系统发育树.结果表明,在长期稳定运行后不同MBR中形成了各自特有的生态群落,进水水质对总细菌的群落结构有着较大的影响,处理含有较复杂成分废水的反应器中,种群多样性较高,Shannon指数分别为0.77和0.78.总细菌中,主要优势种群以Proteobacteria纲(8个OUTs)和Bacillus属(2个OUTs)为主.不同反应器中氨氧化菌群结构较为相似,存在着相同的顶级优势群落;测序结果表明MBR中存在着多种亚硝化菌属,其中以亚硝化单胞菌属最为普遍,并鉴定出2株反硝化菌属UncuItured Achromobacter sp.和Uncultured denitrifying bacterium,说明反应器中可能同时存在多种硝化和脱氮途径.  相似文献   

5.
秦宇  郭劲松  方芳  杨国红 《环境科学》2009,30(2):493-498
为研究溶解氧及曝停比对单级自养脱氮系统微生物群落结构的影响,从不同溶解氧水平及曝停比条件下的SBBR单级自养脱氮反应器中采集活性污泥及生物膜样品,进行PCR-DGGE及条带统计分析.结果表明,经过1.5 a稳定运行,该系统内微生物群落结构与接种污泥相比已变得简单且较稳定.曝停比为2 h∶2 h的条件下,中高低3种溶解氧水平中,生物膜微生物群落丰富度值均高于活性污泥.DO在(曝气)2.0 mg/L(停曝) 0.4 mg/L时系统运行效能最佳,微生物群落丰富度值最高,生物膜和活性污泥样品中条带数分别约为14条和10条,微生物的多样性及相互协同代谢过程是维持单级自养脱氮系统具有较高运行效能的一个关键因素.此外,曝停比对单级自养脱氮系统微生物群落结构有较大影响.3 h∶5 h的较长曝停周期下,活性污泥与生物膜微生物组成接近,相似性为100%,各类细菌虽在活性污泥与生物膜中均能生存但活性较低,系统运行效能差.  相似文献   

6.
针对课题组开发的两相一体式污泥浓缩消化反应器(TISTD),采用16S rDNA、PCR-TGGE等分子生物学技术,对反应器在不同阶段稳态下的内、外反应室中微生物的多样性和种群结构进行了研究.结果表明:TISTD反应器内外反应室中微生物群落均呈现高度的多样性分布,优势菌群明显,群落结构差异性很大,且外反应室较内反应室具有更多的微生物种群数量,外室是水解酸化的场所,污泥在外室经过微生物的水解酸化后进入内室,内室是严格厌氧的产甲烷场所;从反应器运行的不同时期来看,反应器中微生物优势种群呈现一定的差异,随着反应器负荷的改变,微生物的群落结构和种群数量也呈现出明显的演替过程,优势种群的功能地位处在动态变化中.微生物种群之间通过协同和竞争作用,形成了特定生态位的群落结构.研究结果解释了TISTD反应器同时浓缩和消化的工作机理,并为其推广、应用及优化调控提供了理论基础.  相似文献   

7.
通过运行双室的异养-硫自养联合反应器(下部为异养区、上部为自养区),去除高浓度高氯酸盐(ClO_4~-)废水.考察不同进水碳氯比(C/Cl)和ClO_4~-浓度条件下,ClO_4~-的降解特性及出水硫酸盐(SO_4~(2-))浓度.结果表明,通过调节进水C/Cl比由2~1,反应器在36 d快速成功启动;进水ClO_4~-浓度由250 mg·L~(-1)提升至400 mg·L~(-1)时,微生物对高浓度ClO_4~-表现出良好的耐受性,ClO_4~-的去除率始终稳定在95%以上;通过调整C/Cl比至1.2,可减少自养区ClO_4~-负荷,使出水SO_4~(2-)浓度降至250 mg·L~(-1)以下.此外,本研究证实了细菌分泌的溶解性有机物中色氨酸和酪氨酸是造成自养区出水TOC升高的原因.由于采用异养-硫自养联合工艺,控制碳源的投加,从而减少了异养区污泥的产量;同时,异养区产生碱度可以平衡自养区产生的酸度,减少了自养过程中碱度的投加;异养区产生碳酸盐也可以作为自养区的碳源,实现了异养区和自养区功能上的"互补".  相似文献   

8.
为研究低温条件下曝气生物滤池(BAF)处理高盐贫营养含氮废水的启动特性,在系统运行水力停留时间为1h,水温为12~16℃,气水比为2:1(溶解氧控制在3~5mg/L),进水p H值为7.38~8.23,高锰酸盐指数为5.11~9.46mg/L,氨氮为3.27~4.88mg/L的工况条件下,对比考察了接种普通活性污泥和海水底泥沸石BAF处理人工模拟海水养殖废水的低温启动规律和微生物种群特性.结果表明,接种海水底泥与普通活性污泥的BAF反应器分别于39和35d完成挂膜启动,氨氮平均去除率分别为91%、95.7%,出水氨氮平均浓度分别为0.36、0.17mg/L;高锰酸盐指数的平均去除率分别为31.9%、36.5%,出水平均浓度分别为4.96、4.63mg/L.变性梯度凝胶电泳(DGGE)和16Sr DNA基因测序分析发现,两座反应器微生物种群结构发生了较大变化,普通活性污泥、海水底泥及其对应接种种泥的BAF反应器运行稳定一个月后的反冲洗泥样多样性指数分别为2.41、2.63、2.88、2.65;随着运行时间的延长,两种接种方式BAF的微生物种群数量及丰度差异逐渐减弱,优势种群趋于一致,表现出很强的相似性;部分耐盐的氨氧化菌成为优势菌种,稳定运行的BAF生物种群以γ-变形菌纲为主.  相似文献   

9.
采用PCR—DGGE技术,对降解PCP厌氧生物反应器污泥颗粒化过程微生物种群结构的时空分布变化特性进行了初步研究.结果表明,随着PCP负荷增加与污泥的颗粒化,污泥真细菌、古细菌组成均发生明显变化.当反应器稳定运行时,反应器内上下层污泥微生物种群结构组成相似,真细菌相似性Cs在71.4~77.6之间,古细菌Cs在70.4~76.7之间.反应器一旦受污染物负荷冲击干扰,反应器上下层污泥真细菌组成相似性Cs降至43.3,古细菌降至35.2.测序表明,颗粒污泥中存在不可培养的脱氯细菌,优势产甲烷细菌为Methanosaeta concilii、Methanobacterium sp.、Methanobrevibacter sp.和Methanothfix soehngenii.  相似文献   

10.
序批式生物反应器填埋场脱氮微生物多样性分析   总被引:7,自引:6,他引:1       下载免费PDF全文
为探究序批式生物反应器填埋场脱氮过程中的微生物作用机制,本研究采用建立脱氮功能基因(amoA、nosZ)克隆文库及PCR-RFLP技术对序批式生物反应器填埋场垃圾稳定化后期的主要脱氮功能微生物多样性进行分析.结果表明,矿化垃圾反应器中检测到的氨氧化细菌存在高度多样性,大部分为未知类群,且均为不可培养菌或未经分离获得的细菌,经系统发育树分析系统内氨氧化细菌以β-变形菌门中的亚硝化单胞菌属(Nitrosomonas)为主;新鲜垃圾反应器中反硝化细菌种群丰富,主要有β-变形菌纲中的陶厄氏菌属(Thauera)和硫杆菌属(Thiobacillus).Thauera属在好氧条件下具有反硝化特性,Thiobacillus属中的脱氮硫杆菌(Thiobacillus denitrificans)是一种硫自养反硝化菌,可见新鲜垃圾单元稳定化后期以好氧反硝化和自养反硝化的脱氮途径为主.此外文库中检测到的一部分反硝化细菌可能归属于α-变形菌纲的慢生根瘤菌科(Bradyrhizobiaceae).  相似文献   

11.
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.  相似文献   

12.
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.  相似文献   

13.
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.  相似文献   

14.
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.  相似文献   

15.
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.  相似文献   

16.
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.  相似文献   

17.
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.  相似文献   

18.
The potential harm of heavy metals is a primary concern in application of sludge to the agricultural land. A pot experiment was conducted to evaluate the effect of two sludges on fractionation of Zn and Cu in soil and their phytotoxicity to pakchoi. The loamy soil was mixed with 0%, 20%, 40%, 60% and 80% (by weight) of digested sewage sludge (SS) and composted sludge (SC). The additions of both sludges caused a significant raise in all fractions, resulting in that exchangeable (EXCH) and organic bound (OM) became predominance of Zn and organic bound Cu occupied the largest portion. There was more available amount of Zn and Cu in SS treatments than SC treatments. During the pot experiment, the concentration of Zn in EXCH, carbonate (CAR) and OM and Cu in EXCH and OM fractions decreased in all treatments, so their bioavailability reduced. Germination rate and plant biomass decreased when the addition rate was high and the best yield appeared in 20% mixtures at the harvest of pakchoi. The two sludges increased tissue contents of Zn and Cu especially in the SS treatments. Zn in pakchoi was not only in relationship to ΔEXCH and ΔCAR forms but also in ΔOM forms in the sludge-soil mixtures. Tissue content of Cu in pakchoi grown on SC-soils could not be predicted by ΔEXCH. These correlation rates between Zn and Cu accumulation in pakchoi and variation of different fractions increased with time, which might indicate that sludges represented stronger impacts on the plant in long-term land application.  相似文献   

19.
A hydroponic experiment was carried out to study intraspecific differences in the effects of different concentrations of cadmium (Cd)(0-10 mg/L) and arsenate (As(V)) (0-8 mg/L) on the growth parameters and accumulation of Cd and As in six wheat varieties Jing-9428, Duokang-1, Jingdong-11, Jing-411, Jingdong-8 and Zhongmai-8. The endpoints of wheat seedlings, including seed germination,biomass, root length and shoot height, decreased with increasing the Cd and As concentrations. Significant differences in seed germination, biomass, root length, shoot height and the accumulation of Cd and As were observed between the treatments and among the varieties (p < 0.05). The lethal dosage 50% were about 20, 80, 60, 60, 80 and 20 mg As/L for Jing-9428, Duokang-1, Jingdong-11,Jing-411, Jingdong-8 and Zhongmai-8, respectively, and the corresponding values for Cd were about 30, 80, 20, 40, 60 and 10 mg Cd/L, respectively. Among the six varieties, Duokang-1 was found to be the most resistant to Cd and As toxicity, and Zhongmai-8 was the most sensitive to Cd and As co-contamination. The resistance of the six varieties was found dependant on the seedling uptake of Cd and As. Duokang-1 was the most suitable for cultivation in Cd and As co-contaminated soils.  相似文献   

20.
Decomposition of alachlor by ozonation and its mechanism   总被引:1,自引:0,他引:1  
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.  相似文献   

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