一组中温厌氧消化菌群高温启动过程中的菌群多样性变化研究

该研究利用37℃培养的中温厌氧消化菌群为菌源,直接提温至50℃驯化培养,获得连续处理高浓度糖蜜废水的厌氧消化产甲烷菌群,并考察高温驯化过程中菌群结构及多样性的变化特征.结果表明,中温厌氧消化菌群直接转入高温培养后,在高浓度有机废水连续进料的条件下,厌氧消化过程能够快速启动生成甲烷,并在22 d后形成稳定的高温厌氧消化产甲烷菌群,平均甲烷生成效率为162.3 m L CH_4/g COD.乙酸和丙酸是厌氧发酵液内的2类主要有机酸,产气稳定期间的质量浓度分别为25.3和145.3 mg·L~(-1).转入高温培养后,菌群结构产生巨大变化,细菌变异程度强于古菌,并逐渐稳定成为以代谢糖、多种有机酸的细菌和产甲烷古菌为主要优势菌群的高温厌氧消化菌群.克隆结果显示细菌菌群以Thermacetogenium和Acetomicrobium faecal为主要优势菌群,分别占细菌克隆文库的33.44%和20.99%;古菌菌群以Methanosaeta和Methanoculleus为主要优势菌群,占古菌克隆文库的56.40%和39.75%.转入高温培养后,产甲烷古菌的总生物量下降,含量约为7.6×106拷贝/g活性污泥.研究结果对阐明温度选择压力对厌氧消化菌群结构与功能影响,改进高温厌氧消化菌群富集方法具有重要意义.     

多相内循环厌氧反应器内颗粒污泥特性分析

该文对处理柠檬酸废水的多相内循环厌氧反应器中颗粒污泥的理化特性和微生物特性进行了实验研究。扫描电镜下厌氧颗粒污泥呈黑色、球形、表面光滑且有空隙;筛分法得到90%的污泥粒径分布在0.45~2.0 mm范围内;重力沉降法得到90%以上颗粒污泥的沉降速度大于50 m/h,沉降性能良好;高温灼烧法测得颗粒污泥的挥发分占总固体的64.56%,有机物含量高。采用454高通量测序的方法对厌氧颗粒污泥中的微生物进行鉴别,细菌中共有约30个菌门和100个菌属,其中Anaerofustis、Anaerolineae、Spirochaeta和Longilinea是优势菌属,分别占细菌总量的4.91%、4.78%、4.41%、3.98%。这些细菌的共同特征是可以将大分子有机物水解并转化成酸。古生菌中广古菌门(Euryarchaeota)占92.48%,其中甲烷鬃菌属(Methanosaeta)占7.81%,甲烷杆菌属(Methanobacterium)占49.35%,主要作用是产甲烷。微生物的多样性指数较高,表明厌氧颗粒污泥中微生物种类丰富,有利于厌氧反应器的稳定运行。     

高通量测序研究酒精废水治理中厌氧活性污泥的微生物菌群

厌氧消化是治理酒精生产废水的主要技术,本实验基于Illumina Mi Seq高通量测序研究了厌氧活性污泥的微生物菌群,在属(genus)水平上分析了污泥中细菌和古菌的种类和相对丰度,探讨了细菌和古菌的功能.结果表明:高温厌氧活性污泥(TAS)中相对丰度≥1.0%的细菌属有10个,包括Coprothermobacter、Longilinea、Levilinea等,它们的主要功能是分解蛋白质、代谢碳水化合物生成有机酸、乙酸氧化、硫酸盐还原.中温厌氧活性污泥(MAS)中相对丰度≥1.0%的细菌属有21个,包括Acinetobacter、Succinibibrio、Meniscus、Longilinea等,它们的主要功能是代谢碳水化合物生成有机酸、脂肪酸和有机酸氧化、分解结构顽固化合物.TAS中古菌属主要是产甲烷古菌Methanosaeta、Methanobacterium和Methanothermbacter,它们的相对丰度分别为57.72%,28.35%和9.90%.MAS中古菌属主要是产甲烷古菌Methanobacterium、Methanosarcina、Methanosaeta和Methanosphaera,它们的相对丰度分别为33.21%、23.70%、16.63%和11.77%.高通量测序研究充分展示了厌氧活性污泥的微生物菌群,可为阐释酒精废水治理中厌氧消化技术的生物机制及其技术改良提供参考.     

高效厌氧产甲烷颗粒污泥中微生物多样性及定量化研究

以小试高效厌氧反应器不同运行阶段的颗粒污泥为对象,利用变性梯度凝胶电泳(DGGE)、荧光原位杂交(FISH)和实时定量链式聚合酶反应(RTQ-PCR)等技术研究了其中微生物种群的多样性、特征微生物的空间分布和定量关系,结果表明:随着反应器有机负荷逐渐提高,颗粒污泥中古菌的群落结构的变化较细菌更为明显;细菌多分布在颗粒外层,而古菌则主要分布在颗粒内层;古菌含量略少于细菌,但有逐渐增多的趋势;产甲烷丝菌在古菌中的含量增加明显.     

阿维菌素废水工业化UASB颗粒污泥产甲烷菌群分析

产甲烷菌群是厌氧颗粒污泥的重要功能菌群,采用荧光原位杂交(fluorescencein situhybridization,FISH)技术对阿维菌素废水处理工业化UASB颗粒污泥产甲烷菌群进行分析.结果表明,不同形成阶段(不同粒径)颗粒污泥表面和内部剖面,一般产甲烷菌(methanogens)、产甲烷杆菌(Methanobacteriales)和产甲烷八叠球菌(Methanosarcinales)的分布形态相同,但相对丰度存在差异.颗粒污泥内部剖面产甲烷菌群相对丰度大于表面;产甲烷杆菌相对丰度大于产甲烷八叠球菌;颗粒污泥表面和内部剖面产甲烷菌群相对丰度范围为(25.50±8.63)%~(48.67±8.87)%;1.0~2.0 mm粒径颗粒污泥产甲烷菌群相对丰度最大,内部剖面和表面一般产甲烷菌的相对丰度分别为(47.08±8.26)%和(48.67±8.87)%.废水阿维菌素残留可能对产甲烷菌群具有抑制作用.不同粒径颗粒污泥最大比产甲烷活性范围为1.311~1.562 g/(g.d),变化趋势与COD去除率变化趋势相同,而且与产甲烷菌群相对丰度变化趋势一致,表明颗粒污泥生物活性与产甲烷菌群密切相关.     

序批式接触氧化反应器中细菌多样性及其功能

采用分子生物学手段,通过构建16S rDNA基因文库,对新型剩余污泥减量化处理系统——序批式生物砾间接触氧化反应器中细菌多样性进行了系统发育分析,并讨论了多种细菌共存对剩余污泥减量化的贡献.共有72个克隆子用于细菌系统发育分析. 结果表明:填料表面附着细菌与孔隙内细菌由变型菌属、噬纤维菌-屈挠杆菌-拟杆菌组(CFB)、硝化杆菌科、低G+C革兰氏阳性细菌、高G+C革兰氏阳性细菌、疣微菌科和绿菌科等七大类细菌组成. 其中,优势菌群分别是以兼具呼吸/发酵代谢方式的β变形菌纲(分别占生物膜和内泥中克隆子总量的18%和35%)和δ变形菌纲,以及以呼吸/发酵为主要代谢方式的CFB(分别占克隆子总量的24%和23%). 多种细菌对剩余污泥减量化的主要功能可归纳为能量解偶联、共代谢作用、生物溶胞作用和慢性生长种群的影响.      

16S rDNA克隆文库方法分析好氧颗粒污泥细菌组成

采用构建16S rDNA克隆文库方法对好氧颗粒污泥的细菌种群多样性进行研究. 随机测定了82个克隆子序列(700 bp),Blast比对结果表明,好氧颗粒污泥中微生物群落具有高度多样性,可分为7个主要类群,其中,β变形菌(β-Proteobacteria)类群和鞘脂杆菌(Sphingobacteria)类群在文库中所占比例最大,分别为34.16%和30.50%;其次是Candidate division TM7类群、黄杆菌(Flavobacteria)类群和γ变形菌(γ-Proteobacteria)类群,分别为9.76%,7.32%和7.32%;放线菌(Actinobacteria)类群和α变形菌(α-Proteobacteria)类群所占比例相对较小,分别为4.88%和1.22%. 序列分析结果表明,好氧颗粒污泥中不仅含有对好氧颗粒污泥形成和稳定运行具有重要作用的食酸菌属(Acidovorax)细菌、假单胞菌(Pseudomonas)等细菌,还含有对CODCr和氨氮具有很好去除能力的Micropruina glycogenica,丛毛单胞菌科(Comamonadaceae)等细菌.      

活性炭在中高温条件下对玉米秸秆厌氧发酵的影响

为提高玉米秸秆厌氧发酵的产气效果,本文研究活性炭在中温(38℃)和高温(50℃)条件下对玉米秸秆厌氧发酵产甲烷的效果及微生物学机制.结果表明,添加活性炭能显著促进秸秆厌氧发酵产甲烷,中、高温试验组(添加活性炭)的累积产甲烷量分别比对照提高了63%和96%;DGGE的结果显示,高温试验组(添加活性炭)和对照组(未添加活性炭)的发酵液中的优势细菌菌群分别是Clostridiale bacterium和Bacillus,中温对照组发酵液和中温试验组发酵液未发现明显优势菌种.添加活性炭分别有利于氢营养型的甲烷鬃毛菌(Methanosaeta concilii strain)和乙酸营养型的醋酸甲烷八叠球菌(Methanosarcina acetivorans strain),在中温、高温试验组的发酵液中形成优势古菌菌群.中温试验组活性炭载体上的优势古菌菌群为甲烷鬃毛菌(Methanosaeta concilii strain),而高温试验组活性炭载体上的优势古菌菌群主要为甲烷八叠球菌嗜高温菌属(Methanosarcina thermophila strain).     

高钙废水颗粒污泥中古菌菌群结构变化的分析

为了解高浓度Ca~(2+)与颗粒污泥中古菌菌群结构的关系,利用高通量测序分析IC(Internal Circulation)反应器(以废纸造纸废水为处理对象)内不同高度处颗粒污泥中古菌菌群组成,并利用SEM(Scanning Electron Microscope)和EDS(Energy Dispersive X-ray Spectroscopy)分析颗粒污泥的形态结构和元素组成,同时借助红外光谱和XRD(X-ray diffraction)分析不同高度处颗粒污泥化学结构中各官能团的分布情况和无机成分.结果表明,随着反应器高度的增加,颗粒污泥中的无机成分CaCO_3也随着增加,而种泥中钙含量只有1.81%.种泥和ICR反应器内的颗粒污泥中古菌在门分类水平上主要包括Euryarchaeota和Bathyarchaeota,两者之和占90%以上.在属分类水平上,主要包括Methanosaeta、Methanoregula、Methanobacterium、Methanosarcina、Methanolinea、Methanospirillum等.种泥中古菌的物种多样性要明显高于IC反应器中,在IC反应器中不同高度处古菌生物多样性由高到低的顺序是5 m2 m7 m,5 m处的古菌物种更加丰富.随着反应器高度的增加,乙酸营养型甲烷菌的相对丰度呈现递增趋势,氢营养型甲烷菌则呈现递减趋势,而种泥中氢营养型甲烷菌相对丰度较大.这表明颗粒污泥在高钙废水的长期作用下,氢营养型甲烷菌的相对丰度减少,乙酸营养型甲烷菌的相对丰度增多,系统中古菌物种的相对丰度和多样性有所下降.CaCO_3沉淀使颗粒污泥的产甲烷活性降低,主要表现为其对氢营养型甲烷菌具有较大的抑制作用.     

污泥性质对微波预处理-厌氧消化的影响及古菌群落结构分析

污泥性质对预处理强化厌氧消化工艺的效果有较大影响.本研究对比了某实际污水厂A~2O工艺和A~2O-MBR工艺产生的剩余污泥在微波预处理-厌氧消化过程中污泥性质的变化与产气效果,并考察了在预处理和厌氧消化过程中污泥的古菌群落结构变化.结果表明,A~2O工艺剩余污泥有机质含量比A~2O-MBR污泥高出16. 4%(分别为66. 4%和50. 0%),SCOD、溶解性蛋白质和多糖分别为后者1. 24、2. 02和4. 84倍,具有更好的可生物降解性.虽然预处理对生物降解性差的A~2O-MBR污泥有机物释放效果更好,但A~2O污泥在微波预处理-厌氧消化后产甲烷量比相应处理后的A~2O-MBR污泥多26. 1%.两种剩余污泥的古菌群落结构差异较大,A~2O-MBR污泥中甲烷丝菌属和甲烷八叠球菌属丰度分别比A~2O污泥多3. 68%和19. 73%.预处理对古菌群落的丰富度和均匀度影响相对较小,但厌氧消化后波动较大.污泥中有机组分不同是引起古菌群落结构变化的重要影响因素.     

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.     

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.     

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.     

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.