厌氧型甲烷氧化菌降解煤吸附甲烷实验研究

以贵州六枝特区龙岭煤矿大巷中泥样作为厌氧型甲烷氧化菌富集源,以甲烷作为培养过程中唯一碳源,从中筛选出可以在低氧(1.99%)或无氧条件下对甲烷具有较高降解效能的菌种,并自主开发出甲烷氧化菌降解煤吸附甲烷实验分析系统。实验结果表明,在压力为1~5 MPa范围内无论是低氧或无氧状况下,甲烷压力越大越有利于其降解;稀氧条件下煤样对甲烷的吸附量相对于纯甲烷气体吸附量有所降低,然而在同等压力条件下稀氧环境中二氧化碳的增加量及甲烷的降解率都要明显大于无氧条件下,低氧状况下甲烷的最高降解率为47%,最大二氧化碳生成量可达40 cm^3。     

Characterization of Methylocystis strain JTA1 isolated from aged refuse and its tolerance to chloroform

To accelerate the efficiency of methane biodegradation in landfills,a Gram-negative,rod-shaped,non-motile,non-spore-forming bacterium,JTA1,which can utilize methane as well as acetate,was isolated from the Laogang MSW landfills,Shanghai,China.Strain JTA1 was a member of genus Methylocystis on the basis of 16S rRNA and pmoA gene sequence similarity.The maximum specific cell growth rates(μmax=0.042 hr-1,R2=0.995) was derived through Boltzmann simulation,and the apparent half-saturation constants(Km(app)=7.08mmol/L,R2=0.982) was calculated according to Michaelis-Menton hyperbolic model,indicating that Methylocystis strain JTA1 had higher-affinity potential for methane oxidation than other reported methanotrophs.By way of adding the strain JTA1 culture,the methane consumption of aged refuse reached 115mL,almost two times of control experiment.In addition,high tolerance of Methylocystis strain JTA1 to chloroform could facilitate the methane oxidation of aged refuse bio-covers.At the chloroform concentration of 50mg/L,the methane-oxidation rate of bio-cover reached 0.114mL/(day·g),much higher than the highest rate,0.0135mL/(day·g),of reported bio-covers.In conclusion,strain JTA1 opens up a new possibility for environmental biotechnology,such as soil or landfills bioremediation and wastewater decontamination.     

3D model for a secondary facultative pond

This paper describes a comprehensive model of wastewater treatment in secondary facultative ponds, which combines 3D hydrodynamics with a mechanistic water quality model. The hydrodynamics are based on the Navier-Stokes equation for incompressible fluids under shallow water and Boussinesq assumptions capturing the flow dynamics along length, breadth and depth of the pond. The water quality sub model is based on the Activated Sludge Model (ASM) concept, describing COD and nutrient removal as function of bacterial growth following Monod kinetics, except for Escherichia coli removal, which was modelled as first order decay. The model was implemented in the Delft3D software and was used to evaluate the effect of wind and the addition of baffles on the water flow pattern, temperature profiles in the pond and treatment efficiency. In contrast to earlier models reported in the literature, our simulation results did not show any significant improvement in COD removal (based on the ASM concept) with addition of baffles or under intermittent wind-induced mixing. However, E. coli removal efficiency, based on a first order decay approach, showed a fair improvement in the presence of baffles or intermittent wind-induced mixing. Furthermore, simulations with continuous wind effect showed a decrease in removal efficiency for COD but a further increase in E. coli removal efficiency. Such contrasting results for two different approaches in modelling could indicate that the first order decay concept might not be appropriate to describe all the interactions between biochemical processes in a pond. However, these interpretations remain theoretical, as the model needs validation with field data.     

Diversity of methanotrophs in Zoige wetland soils under both anaerobic and aerobic conditions

Zoige wetland is one of the most important methane emission centers in China. The oxidation of methane in the wetland a ects global warming, soil ecology and atmospheric chemistry. Despite their global significance, microorganisms that consume methane in Zoige wetland remain poorly characterized. In this study, we investigated methanotrophs diversity in soil samples from both anaerobic site and aerobic site in Zoige wetland using pmoA gene as a molecular marker. The cloning library was constructed according to the pmoA sequences detected. Four clusters of methanotrophs were detected. The phylogenetic tree showed that all four clusters detected were a liated to type I methanotrophs. Two novel clusters (cluster 1, cluster 2) were found to relate to none of the recognized genera of methanotrophs. These clusters have no cultured representatives and reveal an ecological adaptation of particular uncultured methanotrophs in Zoige wetland. Two clusters were belonging to Methylobacter and Methylococcus separately. Denaturing gradient gel electrophoresis gel bands pattern retrieved from these two samples revealed that the community compositions of anaerobic soil and aerobic soil were di erent from each other while anaerobic soil showed a higher metanotrophs diversity. Real-time PCR assays of the two samples demonstrated that aerobic soil sample in Zoige wetland was 1.5 times as much copy numbers as anaerobic soil. These data illustrated that methanotrophs are a group of microorganisms influence the methane consumption in Zoige wetland.     

覆盖层甲烷氧化动力学和甲烷氧化菌群落结构

基于实际填埋场覆盖土建立了可实时在线监测生物气的模拟覆盖层系统,连续监测了不同垂直梯度生物气浓度变化.甲烷通量不变时,覆盖层内不同梯度生物气浓度基本保持不变,系统持续稳定运行,甲烷通量变化后2~3 h生物气可再次持续稳定.考察了覆盖层甲烷氧化特性与甲烷通量的关系,深度大于20 cm,氧气浓度随甲烷通量的增大呈减小趋势,表层氧气浓度与甲烷通量无相关性,不同梯度的甲烷氧化速率与甲烷通量呈正相关(R2变化范围0.851~0.999).为避免覆盖土脱离系统环境造成的误差,以动态连续监测结果为基础,利用双基质Michaelis-Menten方程拟合了覆盖层甲烷氧化动力学(R2范围为0.902~0.955),得到覆盖土半饱和常数Km为0.157~0.729,Km随深度的增加而增大.利用高通量测序技术分析了原始覆盖土和经模拟覆盖层运行后的甲烷氧化菌群落结构,运行后甲烷氧化菌OUT数量显著增多,优势菌群为Ⅰ型菌的Methylobacter和Methylophilaceae及Ⅱ型菌Methylocystis.     

内蒙古高原干涸湖泊反硝化及甲烷氧化细菌的群落分析

以内蒙古高原半干旱草原区干涸湖泊为研究对象,应用末端限制性片段长度多态性(T-RFLP)技术对湿地土壤中的反硝化和甲烷氧化关联菌群的关键功能基因nosZ和pmoA进行了研究,根据末端限制性片段(T-RFs)分析了两种菌群的群落结构及多样性变化,以及环境因素所产生的影响,并探讨了土壤微生物群落对湖泊干涸的响应机制.结果表明:干涸湖泊的湖心、湖底土壤反硝化和甲烷氧化细菌多样性指数较高,群落结构较复杂;而较早干涸的湖坡以及相邻草原对照区,两个菌群多样性指数较低,群落结构相对单一.从湖心到相邻草原,反硝化和甲烷氧化细菌的群落相似度逐渐降低,显示两个菌群处于动态演替中.湖泊干涸过程中,水分含量和有机质含量对反硝化和甲烷氧化细菌的群落结构影响显著;高pH值和高铵态氮分别对反硝化和甲烷氧化细菌有抑制作用.随着湖泊干涸,土壤微生物群落因环境扰动而受到胁迫,这一过程会加剧CO2等温室气体的排放.     

松嫩平原芦苇湿地退化与修复过程中土壤细菌和甲烷代谢微生物的群落结构

湿地是全球CH₄重要的源与汇.受人为活动和气候条件影响,我国湿地退化严重,相关部门近年来已逐步开展湿地生态修复的工作.为研究湿地退化与修复过程中细菌和甲烷代谢微生物群落结构的响应,以松嫩平原芦苇湿地为研究对象,采集原始未退化芦苇湿地土壤、退化的和正在修复的芦苇湿地土壤,采用基于细菌16S rRNA基因、产甲烷菌mcrA基因和甲烷氧化菌pmoA基因的高通量测序技术研究细菌和甲烷代谢微生物的多样性和群落组成.结果表明,芦苇湿地退化导致土壤细菌和产甲烷菌的α多样性降低,甲烷氧化菌的α多样性升高,而细菌和产甲烷菌的α多样性与土壤含水率呈显著正相关关系,含水率越高的湿地土壤产甲烷菌的多样性也越高.原始未退化芦苇湿地土壤中细菌Rhizobiales和产甲烷菌Methanobacteriaceae的相对丰度较高;湿地退化导致根际促生菌Rhizobiales的相对丰度下降,致病菌Burkholderiaceae、耐污染细菌Sphingomonas、抗辐射细菌Rubrobacter以及Type Ⅰ型耐受极端环境的甲烷好氧氧化菌Methylobacter、Methylomonas和Methylococcus的相对丰度上升;正在修复的芦苇湿地土壤中细菌Bacillus和产甲烷菌Methanosarcinaceae、Methanomicrobiaceae以及Type Ⅱ型甲烷好氧氧化菌Methylocystis的相对丰度较高.因此,不同的芦苇湿地状态可以间接改变土壤性状进而改变湿地甲烷代谢菌群落结构.     

水稻土甲烷氧化菌对镉胁迫的响应

重金属污染影响土壤微生物群落结构与活性,间接影响土壤碳（如CO2、CH4）的生物地球化学循环和全球气候变化。甲烷氧化菌氧化消耗CH4,降低大气中CH4含量,在缓解由温室气体导致的全球温暖化方面起着重要作用。本研究通过短期土壤培养实验,比较研究了不同强度重金属镉（Cd）胁迫下,水稻土中甲烷氧化菌的多度、群落组成及其氧化CH4潜势的差异。结果表明,添加Cd含量越大,水稻土氧化CH4潜势越弱,甲烷氧化菌pmoA基因拷贝数显著减少;甲烷氧化菌多度与水稻土氧化CH4潜势之间存在显著正相关关系（P〈0.001）。群落组成分析发现,在相对低含量Cd（1 mg.kg-1）条件下,有新的甲烷氧化菌菌属出现,而添加较高含量Cd（10 mg.kg-1）时甲烷氧化菌种类减少。总之,Cd胁迫降低水稻土中甲烷氧化菌多样性及其氧化CH4潜势,可能导致原位CH4消耗减少,从而增加稻田CH4排放。     

基于甲烷氧化菌的城镇污水厂尾水极限脱氮系统构建及机制

好氧甲烷耦合反硝化(AME-D)在城镇污水厂尾水深度脱氮方面具有巨大的应用潜力,研究采用改良型反硝化生物滤池,利用低浓度甲烷构建出AME-D极限脱氮系统.研究发现该系统在间歇式运行方式下,出水中总氮和氨氮的平均浓度能达到1.05 mg·L^-1和0.54 mg·L^-1,其平均去除率分别为94.77%和93.30%.拉曼光谱分析结果显示,由NO^-₃对称伸缩引起的峰明显消失,由醇COH面外弯曲或C—H面外弯曲振动吸收引起峰明显增强,甲烷被氧化形成的中间产物可能主要为醇类物质. 16S rRNA基因测序结果表明,系统中的甲烷氧化菌主要为Methylocystis(0.27%)、Methylosarcina(0.10%)和Methyloparacoccus(0.12%),反硝化菌主要为Pseudomonas(56.92%)、Paenibacillus(3.52%)和Lysinibacillus(3.00%),硝化菌主要为Nitrospira(0.1%),说明AME-D极限脱氮系统的脱氮功能是由好氧甲烷氧化菌、...     

Nitrifying and denitrifying bacteria in aerobic granules formed in sequencing batch airlift reactors

The purpose of this study was to investigate nitrifying bacteria and denitrifying bacteria isolated from aerobic granules. Aerobic granules were formed in an internal-circulate sequencing batch airlift reactor (SBAR) and biodegradation of NH₃ ⁻-N was analyzed in the reactor. Bacteria were isolated and determined from aerobic granules using selected media. The growth properties and morphology of bacteria colonies were observed by controlling aerobic or anaerobic conditions in the culture medium. It was found that bacteria in aerobic granules were diverse and some of them were facultative aerobes. The diversity of bacteria in aerobic granules was a premise of simultaneous nitrification and denitrification. Translated from China Environmental Science, 2005, 25(2): 218–221 [译自: 中国环境科学]