好氧-沉淀-厌氧工艺剩余污泥减量性能和机理研究

探讨了污泥衰减、能量解偶联、低污泥产率厌氧反应对OSA工艺污泥减量作用的影响.结果表明,污泥衰减是由微生物死亡及吸附于污泥表面的颗粒有机物水解酸化和微生物内源代谢2部分组成.间歇试验污泥厌氧过程中,上清液SCOD、NH 4-N、TP浓度均随厌氧时间逐渐升高,OSA污泥厌氧16 h后溶解性蛋白质高达33.09 mg/L,上升幅度高于多糖浓度的变化,证实了污泥水解现象.OSA污泥内源SOUR可达8 mg/(g·h),是CAS污泥内源SOUR的1.7倍以上,说明OSA系统中较高的内源代谢促使污泥减量.污泥衰减是OSA工艺污泥减量的决定性原因,可占OSA污泥减量效果的66.7%左右.间歇实验证实了OSA系统由于厌氧一好氧耦合环境,存在能量解偶联现象,但由于这种作用引发的污泥减量仅占7.5%左右.OSA工艺污泥厌氧池释放的SCOD作为缺氧反硝化、厌氧释磷、硫酸盐还原及产甲烷的二次基质,由于这些厌氧反应污泥产率低于好氧代谢,使得系统污泥产率下降,约有23.5%的污泥减量是源于这种因素.OSA污泥减量是由多方面因素综合作用的结果.     

铁元素对硫酸盐还原过程的影响及微生物群落响应

采用常规化学分析和微生物群落变性梯度凝胶电泳(DGGE)监测技术,探讨投加不同价态铁元素对硫酸盐还原过程的影响以及相应的微生物群落动态响应.结果表明,反应器启动后5d内硫酸盐去除率达到80%,在此过程中群落条带逐渐减少,但与已分离的硫酸盐还原菌(SRB)菌株一致的条带并没出现;Fe³⁺的投加极显著地改变了原有的高效群落结构,硫酸盐去除率降至20%,氧化还原电位(ORP)有所上升;而Fe²⁺和Fe0的投加未改变已经形成的顶极群落结构模式,也未显著降低硫酸盐去除率,仅硫化物的浓度变化对Fe²⁺的投加有短暂的响应.     

堆肥化过程中微生物群落的动态

通过变性梯度凝胶电泳（DGGE）和平板计数法对堆肥化过程中微生物的区系动态变化进行了研究.结果表明,在堆肥化过程中,微生物数量总的趋势是细菌的数量最多,放线菌次之,真菌的数量最少,中温微生物的数量始终高于高温微生物.当发酵结束后,中温微生物的数量低于发酵初始水平,高温放线菌和高温真菌的数量试验结束后高于初始水平,高温细菌的数量在整个堆肥化过程中变化不大.通过DGGE分析表明,发酵过程中细菌的种类发生了明显的更迭现象.发酵初期Bdellovibrio、Clostridia bacterium、Bacillus、Clostridium等占优势,中期Beta proteobacterium、Petrobacter succinimandens、Nitrospirae bacterium、Clostridium等占优势,后期Clostridium、Beta proteobacterium、Paenibacillus等占优势,而在整个堆肥化过程中Clostridium都是优势种.     

活性污泥厌氧Fe(Ⅲ)还原氨氧化现象初探

采用常规化学分析和微生物群落变性梯度凝胶电泳(DGGE)监测技术,探究了厌氧条件下活性污泥中Fe(Ⅲ)还原氨氧化(Feammox)反应的存在及微生物群落动态响应.结果表明,当反应器运行至第24 d时NH_4~+发生转化,同时检测到NO_3~-和Fe(Ⅱ)的生成,表明活性污泥中存在着Fe(Ⅲ)还原NH_4~+氧化反应,产物主要为NO_3~-和Fe(Ⅱ),并伴随少量N_2生成.经过84d培养,氨氮最大转化量达29.85 mg·L~(-1),转化率为59.7%,出水NO_3~-最高值达24.56 mg·L~(-1).活性污泥中Feammox为产酸过程,体系中p H值下降.整个培养过程中微生物群落条带分布发生变化,参与活性污泥中Feammox反应的部分群落在培养过程获得保留,部分优势菌群获得富集.     

Monitoring impact of mefenacet treatment on soil microbial communities by PCR-DGGE fingerprinting and conventional testing procedures

The effect of acetanilide herbicide mefenacet on soil microbial communities was studied using paddy soil samples with different short-term treatments. The culturable bacteria （plate counts）, dehydrogenase activity and changes in community structure （denaturing gradient gel electrophoresis （DGGE） analysis） were used for biological community assessments. Mefenacet was a significant stimulus to cultural aerobic bacteria and dehydrogenase activity while Sphingobacterium multivorum Y1, a bacterium efficiently degrading the mefenacet, only induced the increasing colony-forming unit （CFU） of bacteria but little effect on dehydrogenase activity during the whole experiment. The degree of similarity between the 16S rDNA profiles of the communities was quantified by numerically analyzing the DGGE band patterns. Similarity dendrograms showed that the microbial community structures of the mefenacet-treated and non-treated soils were not significantly different. But supplement of S. multivorum Y1 could increase the diversity of the microbial community in the mefenacet-polluted paddy soil. This work is a new attempt to apply the S. multivorum Y1 for remediation of the mefenacet-polluted environments.     

接种纤维素降解菌对牛粪堆肥微生物群落的影响

以牛粪和稻草为堆肥原料,利用传统培养法和PCR-DGGE技术相结合对接种纤维素降解菌后堆肥的微生物群落的动态变化进行了分析.结果表明,接种菌堆肥与自然堆肥相比,微生物数量的变化趋势快于自然堆肥,接种菌在堆肥初期能够激发微生物增殖,快速启动堆肥发酵,缩短堆肥进程.接种菌堆肥比自然堆肥的DGGE凝胶图谱条带的差异性大,接种...     

复合菌系BYND-8的种群组成及其对沼气产量的影响

为了明确1组在中温下(30℃)高效分解木质纤维素的复合菌系的菌群组成.研究复合菌系预处理秸秆对沼气发酵的影响,利用平板分离法和变性梯度凝胶电泳(DGGE)法研究了中温木质纤维素降解复合菌系BYND-8的菌种组成多样性,通过添加该复合系菌液到以牛粪为原料的沼气发酵体系,研究了添加秸秆降解液对沼气产量的影响.利用平板法分离...     

农业废物好氧堆肥过程因子对细菌群落结构的影响

采用聚合酶链式反应-变性梯度凝胶电泳(PCR-DGGE)技术研究了农业废物堆肥过程中细菌种群随时间的变化.同时,应用Quantity One2.0和Canoco4.5软件对获得的堆肥细菌种群数据与堆肥过程因子:环境温度、堆体温度、pH、含水率、水溶性有机碳(WSC)、C/N、水溶性氨氮(NH4+-N)和硝氮(NO3--N)进行冗余分析,并做出样点、种群与堆肥过程因子的二维排序图.结果表明,细菌群落(样点)以堆肥过程因子为梯度大体可划分为升温期(1～2d)、高温期(3～11d)、降温期(12～18d)和腐熟期(19～36d)4个阶段,每一阶段均有对应种群存在.不同的堆肥过程因子对细菌种群的影响大小依次为:NO3--N堆体温度WSCC/NNH4+-N含水率pH环境温度,其中,堆体温度、WSC、NO3--N、NH4+-N对细菌种群的影响极显著(p0.01),C/N、pH对细菌种群的影响显著(p0.05),含水率、环境温度对细菌种群的影响不显著.     

应用变性梯度凝胶电泳研究厦门西海域超微型真核浮游生物多样性

应用变性梯度凝胶电泳(DGGE)方法对厦门西海域超微型真核浮游生物的遗传多样性进行周年变化特征研究分析.结果表明,厦门西海域典型测站的超微型真核浮游生物群落具有丰富的多样性组成;厦门西海域超微型真核浮游生物群落遗传结构和主要类群组成季节性变化显著:不同典型测站之间的超微型真核浮游生物群落遗传结构的周年变化特征相似;不同典型测站之间的超微型真核浮游生物群落的类群组成也很相似,由绿藻、Stramenopiles、定鞭金藻、甲藻、未确定类群(unidentified groups)和其他类群组成.     

PCR-DGGE技术解析生物制氢反应器微生物多样性

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 .     

Effect of methamidophos on soil fungi community in microcosms by plate count, DGGE and clone library analysis

Methamidophos was widely used a pesticide in northern China. The potential influences of methamidophos on soil fungal community in black soil were assessed by plate count, 28S rDNA-PCR-DGGE, and clone library analysis. Three methamidophos levels (50, 150, and 250 mg/kg) were tested in soil microcosms. Results from plate count during a 60-d microcosm experiment showed that high concentrations of methamidophos (250 mg/kg) could significantly stimulate fungal populations. DCGE (denaturing gradient gel electrophoresis) fingerprinting patterns showed a significant difference between the responses of culturable and total fungi communities under the stress of methamidophos. Shannon diversity indices calculated from DGGE profiles indicated that culturable fungi in all microcosms with methamidophos treatment increased after 1 week of incubation. However, the diversity indices of total fungi decreased in the first week, as compared to the stimulation of culturable fungi. At the 8th week, however, all the microcosms treated by methamidophos were similar to the control microcosms in community structure as suggested by the Shannon diversity indices for both culturable and total fungi. In contrast, after 1 week the fungal structure of culturable and unculturable both were disturbed to different extent under the stresses of methamidophos by clustering analysis. Clone sequencing analysis indicated the stimulation of pathogenic and unculturable fungal populations by methamidophos treatment, suggetsing potential risks of plant disease outbreak.     

生物造粒流化床污水处理反应器中微生物生长比较分析

利用传统细菌计数方法及分子生物学方法对新近发展起来的生物造粒流化床反应器与A2/O工艺的微生物特性进行比较研究.反应器中好氧菌、反硝化菌及反硫化菌的计数结果显示,单位质量的活性污泥中,生物造粒流化床中好氧菌的分布数是A2/O工艺好氧区的1～2倍,反硝化菌总数介于A2/O工艺的好氧区和缺氧区之间,而反硫化菌的总数介于A2/O工艺的缺氧区和厌氧区之间.用PCR-DGGE技术对生物造粒流化床反应器及A2/O工艺中的微生物进行多样性分析,结果显示,生物造粒流化床反应器的微生物群落比A2/O工艺丰富;DGGE指纹图谱聚类分析表明,生物造粒流化床反应器与A2/O工艺的微生物群落相似性为57.6%.     

高效稳定纤维素分解菌复合系WSC-6的稳定性

对筛选到的一组纤维素分解菌复合系WSC-6,通过变性梯度胶电泳(DGGE)方法研究了菌种的组成稳定性.结果表明,在连续继代培养的第74～83代复合系的菌种组成没有变化,非常稳定.多代继代培养过程中各代的pH值变化趋势一致,pH值从发酵开始的8.7下降到纤维素旺盛分解时的6.5以下;随着分解结束,pH值逐渐恢复到发酵开始时的水平并保持稳定,具有较强的自我调节能力.多代继代培养后复合系各代的滤纸纤维素分解率和CMC糖化差异很小;在发酵液起始pH4～10的范围内,复合系对pH值具有缓冲能力,并正常分解纤维素;经过70～100℃高温处理10min后再转接的复合系对纤维素仍然具有分解能力,功能稳定.     

不同预处理方式对剩余污泥中活菌菌群及ARGs的影响

剩余污泥作为抗生素抗性基因（ARGs）的储库,对其进行预处理有助于后续厌氧消化（AD）,同时控制ARGs进入AD后的传播风险.本文采用叠氮碘化丙锭（PMA）处理手段,来研究不同预处理方式对活菌菌群及携带ARGs的影响.研究结果显示,热碱、热水解和微波预处理对污泥ARGs去除效果较优,分别为3.32log、3.13log和2.95log;超声波预处理对活菌ARGs仅去除0.58log.在热水解预处理时间延长过程中,菌群大量死亡出现在1~2h间,4h后去除率达2.42log.变形菌门是污泥中最主要的优势菌门,厚壁菌门、绿弯菌门在微波、热水解预处理后实现较大提升.相关性结果显示,sulI与tetC高度相似,且同时与多个科属的微生物成显著正相关,推测其宿主范围较广,且sulI与tetC在预处理中较好的去除率说明其宿主菌对预处理耐受性差;ermB、tetA与占比较高的菌群没有正相关.多数ARGs携带菌对热水解、热碱和微波不耐受,污泥预处理能够降低ARGs丰度,适当延长低温热水解预处理时间,才能有效削减污泥中的ARGs.     

MBR和CAS工艺污泥在贫营养培养条件下的微生物群落结构研究

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...     

Characterization of depth-related microbial communities in lake sediment by denaturing gradient gel electrophoresis of amplified 16S rRNA fragments

The characterization of microbial communities of different depth sediment samples was examined by a culture-independent method and compared with physicochemical parameters, those are organic matter （OM）, total nitrogen （TN）, total phosphorus （TP）, pH and redox potential （Eh）. Total genomic DNA was extracted from samples derived from different depths. After they were amplified with the GC-341 f/907r primer sets of partial bacterial 16S rRNA genes, the products were separated by denaturing gradient gel electrophoresis （DGGE）. The profile of DGGE fingerprints of different depth sediment samples revealed that the community structure remained relatively stable along the entire 45 cm sediment core, however, principal-component analysis of DGGE patterns revealed that at greater sediment depths, successional shifts in community structure were evident. The principle coordinates analysis suggested that the bacterial communities along the sediment core could be separated into two groups, which were located 0-20 cm and 21-45 cm, respectively. The sequencing dominant bands demonstrated that the major phylogenetic groups identified by DGGE belonged to Bacillus, Bacterium, Brevibacillus, Exiguobacterium, γ-Proteobacterium, Acinetobacter sp. and some uncultured or unidentified bacteria. The results indicated the existence of highly diverse bacterial community in the lake sediment core.     

Evaluation of a fetus at risk for dihydropteridine reductase deficiency by direct mutation analysis using denaturing gradient gel electrophoresis

Dihydropteridine reductase (DHPR) is an enzyme involved in the recycling of tetrahydrobiopterin (BH₄), which is an obligate co-factor of the aromatic amino acid hydroxylases. DHPR deficiency is a rare, autosomal recessive disorder caused by mutations in the QDPR gene. DHPR-deficient patients are diagnosed by a lack of response to a low phenylalanine diet and by severe neurological symptoms. Final diagnosis is made by measurements of neurotransmitters and pterin metabolites in cerebrospinal fluid (CSF) and urine, in addition to DHPR enzyme activity, which can be assessed in whole red blood cells. Treatment of DHPR deficiency can be difficult and the outcome is not always satisfying, even if all treatment strategies are followed. Therefore prenatal diagnosis is of great importance in affected families. Prenatal diagnosis is possible by measuring DHPR activity in different cell types but this is time consuming. More than 25 different mutations have to date been identified in the QDPR gene and direct identification of a mutation in a fetus would be easy and rapid. We have developed a method based on denaturing gradient gel electrophoresis (DGGE) for the analysis of the QDPR gene. The method is useful for rapid and simultaneous scanning of all exons and flanking intronic sequences of the QDPR gene. We describe the first prenatal diagnosis conducted using this method. Copyright © 2001 John Wiley & Sons, Ltd.     

Methanogenic community dynamics in anaerobic co-digestion of fruit and vegetable waste and food waste

A lab-scale continuously-stirred tank reactor (CSTR), used for anaerobic co-digestion of fruit and vegetable waste (FVW) and food waste (FW) at different mixture ratios, was operated for 178 days at the organic loading rate of 3 kg VS (volatile solids)/(m³.day). The dynamics of the Archaeal community and the correlations between environmental variables and methanogenic community structure were analyzed by polymerase chain reactions - denaturing gradient gel electrophoresis (PCR-DGGE) and redundancy analysis (RDA), respectively. PCR-DGGE results demonstrated that the mixture ratio of FVW to FW altered the community composition of Archaea. As the FVW/FW ratio increased, Methanoculleus, Methanosaeta and Methanosarcina became the predominant methanogens in the community. Redundancy analysis results indicated that the shift of the methanogenic community was significantly correlated with the composition of acidogenic products and methane production yield. Different mixture ratios of substrates led to different compositions of intermediate metabolites, which may affect the methanogenic community. These results suggested that the analysis of microbial communities could be used to diagnose anaerobic processes.     

Response of anaerobes to methyl fluoride, 2-bromoethanesulfonate and hydrogen during acetate degradation

To use the selective inhibition method for quantitative analysis of acetate metabolism in methanogenic systems,the responses of microbial communities and metabolic activities,which were involved in anaerobic degradation of acetate,to the addition of methyl fluoride(CH3F),2-bromoethanesulfonate(BES)and hydrogen were investigated in a thermophilic batch experiment.Both the methanogenic inhibitors,i.e.,CH3F and BES,showed their effectiveness on inhibiting CH4 production,whereas acetate metabolism other than acetoclastic methanogenesis was stimulated by BES,as reflected by the fluctuated acetate concentration.Syntrophic acetate oxidation was thermodynamically blocked by hydrogen(H2),while H2-utilizing reactions as hydrogenotrophic methanogenesis and homoacetogenesis were correspondingly promoted.Results of PCR-DGGE fingerprinting showed that,CH3F did not influence the microbial populations significantly.However,the BES and hydrogen notably altered the bacterial community structures and increased the diversity.BES gradually changed the methanogenic community structure by affecting the existence of different populations to different levels,whilst H2 greatly changed the abundance of different methanogenic populations,and induced growth of new species.     

快速木质纤维素分解菌复合系MC1对秸秆的分解能力及稳定性

以天然水稻秸秆为材料研究了快速降解木质纤维素的细菌复合系MC1对木质纤维素的分解能力;并在不同条件保藏、高温处理以及利用变性梯度胶电泳(DGGE)技术研究了复合系的稳定性.结果表明,复合系MC1在50℃液体静止培养条件下8～10d,把培养液2%干重的水稻秸秆完全分解溶化;经过9d的培养,水稻秸秆的总干重减少81%,其中纤维素减少99%,半纤维素减少74%,木质素减少51%.连续继代培养4a、常温干燥保存4a、-20℃冷冻藏4a、培养液直接在室温和4℃保存1a、90℃处理30min仍具旺盛的分解能力并稳定传代.平板培养基培养证明MC1全部由细菌组成,16SrDNA变性梯度胶电泳(DGGE)检测结果,在6个月内主要条带几乎没有变化,说明MC1的菌种组成相当稳定.MC1对纤维素的分解利用具重要前景.