利用PCR-DGGE技术分析生物陶粒硝化反应器中微生物群落动态

为了揭示生物陶粒硝化反应器中活性污泥的微生物种群多样性,从运行不同时期的反应器中提取活性污泥,利用PCR-DGGE技术初步分析了生物陶粒反应器细菌的种群演替情况.结果表明,随着进水COD值的逐阶段降低,氨氮去除率逐步提高到64.38%.群落结构和优势种群的数量具有时序动态性,微生物多样性与废水的处理效果出现协同变化的特征.测序结果表明,生物陶粒反应器中运行第21天的优势种群除了自养细菌,还有异养细菌存在.其中自养细菌为Nitrosospira.sp和Nitrobacter.sp,异养细菌为Bacillus.sp、Pseudomonas.sp和Pseudochrobactrum.sp.     

Decrease of NH4+-N by bacterioplankton accelerated the removal of cyanobacterial blooms in aerated aquatic ecosystem

We used aerated systems to assess the influence of the bacterioplankton community on cyanobacterial blooms in algae/post-bloom of Lake Taihu, China. Bacterioplankton community diversity was evaluated by polymerase chain reaction-denaturing gradient gel electrophoresis(PCR-DGGE) fingerprinting. Chemical analysis and nitrogen dynamic changes illustrated that NH4+-N was nitrified to NO2-N and NO3-N by bacterioplankton. Finally, NH4+-N was exhausted and NO3-N was denitrified to NO2-N, while the accumulation of NO2-N indicated that bacterioplankton with completely aerobic denitrification ability were lacking in the water samples collected from Lake Taihu. We suggested that adding completely aerobic denitrification bacteria(to denitrify NO2-N to N2)would improve the water quality. PCR-DGGE and sequencing results showed that more than 1/3 of the bacterial species were associated with the removal of nitrogen, and Acidovorax temperans was the dominant one. PCR-DGGE, variation of nitrogen, removal efciencies of chlorophyll-a and canonical correspondence analysis indicated that the bacterioplankton significantly influenced the physiological and biochemical changes of cyanobacteria. Additionally, the unweighted pair-group method with arithmetic means revealed there was no obvious harm to the microecosystem from aeration. The present study demonstrated that bacterioplankton can play crucial roles in aerated ecosystems, which could control the impact of cyanobacterial blooms in eutrophicated fresh water systems.     

蔗渣垫料在零排放发酵床养猪中的应用研究

目前微生物发酵床养猪垫料是选用木屑、谷壳作为原料,来实现零排放养猪。文章选用农业固体废弃物——蔗渣为垫料应用于零排放漏缝发酵床养猪,跟踪了3个月垫料在实际使用中的温度、有机质、氮、磷、钾、微生物指标,对蔗渣的使用效果进行了评价,并建立PCR反应体系来扩增DNA,利用PCR-DGGE技术对垫料中的微生物群落多样性进行研究。结果表明:蔗渣作为垫料,能使垫床温度控制在40⁶5℃之间,保证了良好的发酵效果、且有效活菌数达到0.22×109cfu/g,可以对猪粪便等有机质进行较彻底的消解,因此蔗渣可以作为一种垫料应用于零排放漏缝发酵床;垫料中总养分达到1.371 7%,使用后的垫料可以作为一种原料生产生物有机肥。     

利用PCR-DGGE溯源典型农村塘坝饮用水中的污染

以华南地区典型农村塘坝饮用水为研究对象,采用基于大肠杆菌的两种特异性基因(β-D-葡萄糖苷酶编码基因uidA和膜外周磷通道蛋白编码基因phoE)的群体差异分析(PCR-DGGE),对饮用水中的污染情况进行了溯源研究.结果表明,采用富集培养的混合菌体DNA作为模板扩增大肠杆菌特异性基因更加实用.uidA和phoE的引物均...     

The exposition of a calcareous Mediterranean soil to toxic concentrations of Cr, Cd and Pb produces changes in the microbiota mainly related to differential metal bioavailability

The involvement of the bacterial community of an agricultural Mediterranean calcareous soil in relation to several heavy metals has been studied in microcosms under controlled laboratory conditions. Soil samples were artificially polluted with Cr(VI), Cd(II) and Pb(II) at concentrations ranging from 0.1 to 5000 mg kg⁻¹ and incubated along 28 d. The lowest concentrations with significant effects in soil respirometry were 10 mg kg⁻¹ Cr and 1000 mg kg⁻¹ Cd and Pb. However, only treatments showing more than 40% inhibition of respirometric activity led to significant changes in bacterial composition, as indicated by PCR-DGGE analyses. Presumable Cr- and Cd-resistant bacteria were detected in polluted microcosms, but development of the microbiota was severely impaired at the highest amendments of both metals. Results also showed that bioavailability is an important factor determining the impact of the heavy metals assayed, and even an inverted potential toxicity ranking could be achieved if their soluble fraction is considered instead of the total concentration. Moreover, multiresistant bacteria were isolated from Cr-polluted soil microcosms, some of them showing the capacity to reduce Cr(VI) concentrations between 26% and 84% of the initial value. Potentially useful strains for bioremediation were related to Arthrobacter crystallopoietes, Stenotrophomonas maltophilia and several species of Bacillus.     

间歇运行式生物滴滤池处理油漆废气中试研究

为研究间歇运行式生物滴滤池对油漆生产厂废气净化能力,建立一座中试规模生物滴滤池（BTF）,接种降解菌群,采用8 h/d运行方式净化某油漆厂包装车间废气,并用PCR-DGGE技术揭示BTF细菌群落结构与工艺运行条件间的联系。油漆厂包装车间废气中挥发性有机物（VOCs）主要为甲苯、乙苯、混合二甲苯（间、对和邻二甲苯）,BTF对甲苯、乙苯、混合二甲苯最大去除率分别为88.8%、83.7%和86.3%。DGGE分析显示,BTF稳定运行时,主要优势菌相对丰度较为稳定（F,P>0.05）,其细菌多样性显著低于启动期（F,P>0.05）;同时,下层填料细菌多样性高于上层填料,其细菌结构变化也较上层明显;另外,提升培养液浓度至2倍和4倍对菌群结构亦无显著影响。     

Biological treatment of fish processing wastewater: A case study from Sfax City (Southeastern Tunisia)

The present work presents a study of the biological treatment of fish processing wastewater at salt concentration of 55 g/L. Wastewater was treated by both continuous stirred-tank reactor (CSTR) and membrane bioreactor (MBR) during 50 and 100 days, respectively. These biological processes involved salt-tolerant bacteria from natural hypersaline environments at different organic loading rates (OLRs). The phylogenetic analysis of the corresponding excised DGGE bands has demonstrated that the taxonomic affiliation of the most dominant species includes Halomonadaceae and Flavobacteriaceae families of the Proteobacteria (Gamma-proteobacteria class) and the Bacteroidetes phyla, respectively. The results of MBR were better than those of CSTR in the removal of total organic carbon with efficiencies from 97.9% to 98.6%. Nevertheless, salinity with increasing OLR aggravates fouling that requires more cleaning for a membrane in MBR while leads to deterioration of sludge settleability and effluent quality in CSTR.     

Diversity of methanotrophs in a simulated modified biocover reactor

A simulated landfill biocover microcosm consisting of a modifying ceramsite material and compost were investigated. Results show that the mixture can improve the material porosity and achieve a stable and highly efficient (100%) methane oxidation over an extended operating period. The diversity of the methanotrophic community in the microcosm was assessed. Type I methanotrophs were enhanced in the microcosm due to the increased air diffusion and distribution, whereas the microbial diversity and population density of type II methanotrophs were not significantly affected. Moreover, the type I methanotrophic community structure significantly varied with the reactor height, whereas that of type II methanotrophic communities did not exhibit a spatial variation. Phylogenetic analysis showed that type I methanotroph-based nested polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE) resulted in the detection of eight different populations, most of which are related to Methylobacter sp., whereas that of type II resulted in the detection of nine different populations, most of which are related to Methylocystaceae. Methanotrophic community analysis also indicated that a number of new methanotrophic genera not closely related to any known methanotrophic populations were present.     

梯度负荷下果蔬垃圾厌氧消化性能及微生物群落结构的研究

为优化果蔬垃圾厌氧消化工艺,提高厌氧消化性能,本文通过逐级提高CSTR反应器进料负荷,研究不同负荷下的厌氧消化性能及相应的微生物群落结构变化规律.结果表明,随着进料负荷的增高,容积产气率、甲烷产气量、氨氮、碱度、TCOD、SCOD均逐渐增高,在最高负荷(负荷以VS计)2.50g·L-·1d-1时分别达到最大值:1.22L·L-1d-1,5.10L·d-1,1563.86mg·L-1,7572.23mg·L-1,13283.26mg·L-1,2075.03mg·L-1,甲烷含量及VFA分别稳定在52.46%～54.59%和(879.30±18.69)mg·L-1;同时利用PCR-DGGE技术系统分析了厌氧消化中细菌与古细菌的群落结构,测序结果表明,整个过程中拟杆菌(Bacteroidetes)、甲烷鬃菌(Methanosaeta)及甲烷螺菌(Methanospirillum)为优势微生物,随着负荷的提高,甲烷鬃毛菌(Methanosaeta)活性逐渐降低;聚类分析及主成分分析表明,低负荷条件下(1.50g·L-·1d-1、1.75g·L-·1d-1),微生物种类(细菌、古细菌)差别不明显,且基本处于同一阶段.     

MBR活性污泥培养驯化过程中生物多样性研究

以MBR反应器启动调试阶段的活性污泥为主要研究对象,系统考察了传统活性污泥法(Conventional activated sludge,CAS)污泥接种至MBR反应器内污泥培养驯化过程中生物多样性情况及微生物群落结构的演变规律.同时,在传统污水污泥检测指标的基础上,对各阶段污泥中总细菌基因组DNA进行提取,应用PCR-DGGE分子生物学技术获得了相应的凝胶电泳图谱并进一步分析了菌群间的相似性.结果表明,以CAS污泥为接种污泥在MBR反应器内培养驯化过程中微生物的多样性变化突出,细菌群落结构演替明显,不同阶段菌群间的相似性说明了各阶段菌群的演变关系:污泥培养驯化是一个逐步有序的过程,微生物随反应器内不同时期及环境的变化而调整,逐渐演变成适应MBR工艺的群落结构.