Methanotrophic community structure of aged refuse and its capability for methane bio-oxidation

Aged refuse from waste landfills closed for eight years was examined and found to contain rich methanotrophs capable of biooxidation for methane. Specially, community structure and methane oxidation capability of methanotrophs in the aged refuse were studied. The amount of methanotrophs ranged 61.97 103–632.91 103 cells/g (in dry basis) in aged refuse from Shanghai Laogang Landfill. Type I and II methanotrophs were found in the aged refuse in the presence of sterilized sewage sludge and only Type I methanotrophs were detected in the presence of nitrate minimal salt medium (NMS). The clone sequences of the pmoA gene obtained from the aged refuse were similar to the pmoA gene of Methylobacter, Methylocaldum, and Methylocystis, and two clones were distinct with known genera of Type I methanotrophs according to phylogenetic analysis. Aged refuse enriched with NMS was used for methane biological oxidation and over 93% conversions were obtained.     

Absorption of phosphorus from wastewater by aged refuse excavated from municipal solid waste landfill

Municipal solid waste(refuse) landfill stabilizes as the refuse degrades. After years of biodegradation, the refuse in the landfill ecomes stabilized and aged, which may vary with the local climate, humidity, and composition of refuse placed. In this work, it is found hat the refuse with an age of over 8 years at Shanghai Refuse Landfill has been significantly stabilized and sufficiently aged and is thus uitable for excavation. The 8-year old aged refuse is mechanically screened, and the fine fractions of refuse(aged refuse) with a diameter ess than 2 cm are then used as a biological absorbent for removal of both inorganic and organic phosphorus in livestock wastewater and repared aqueous solution. It is proved that the aged refuse is very effective for the quantitative removal of both types of phosphorus. The bsorption mechanism is proposed. It is considered that phosphorus is firstly absorbed onto the surface of the aged refuse and then used s a substrate for the growth of microorganisms which ultimately leave the aged refuse as sludge.     

Landfill leachate treatment in assisted landfill bioreactor

Landfill is the major disposal route of municipal solid waste（MSW） in most Asian countries. Leachate from landfill presents a strong wastewater that needs intensive treatment before discharge. Direct recycling was proposed as an effective alternative for leachate treatment by taking the landfill as a bioreactor. This process was proved not only considerably reducing the pollution potential of leachate, but also enhancing organic degradation in the landfill. However, as this paper shows, although direct leachate recycling was effective in landfilled MSW with low food waste fraction （3.5%, w/w）, it failed in MSW containing 54% food waste, as normally noted in Asian countries. The initial acid stuck would inhibit methanogenesis to build up, hence strong leachate was yielded from landfill to threaten the quality of receiving water body. We demonstrated the feasibility to use an assisted bioreactor landfill, with a well-decomposed refuse layer as ex-situ anaerobic digester to reducing COD loading in leachate. By doing so, the refuse in simulated landfill column （2.3 m high） could be stabilized in 30 weeks while the COD in leachate reduced by 95%（61000 mg/L to 3000 mg/L）. Meanwhile, the biogas production was considerably enhanced, signaling by the much greater amount and much higher methane content in the biogas.     

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.     

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.     

Evolution on qualities of leachate and landfill gas in the semi-aerobic landfill

To study the characteristics of stabilization in semi-aerobic landfill, large-scale simulated landfill was constructed based on the semiaerobic landfill theory. Consequently, the concentrations of chemical oxygen demand (COD), ammonia nitrogen, and nitrite nitrogen, and the pH value in leachate, as well as the component contents of landfill gas composition (methane, carbon dioxide, and oxygen) in landfill were regularly monitored for 52 weeks. The results showed that COD and ammonia concentrations declined rapidly and did not show the accumulating rule like anaerobic landfill, and remained at about 300 and 100 mg/L, respectively, after 48 weeks. Meanwhile, the descending rate reached 98.9% and 96.9%, respectively. Nitrate concentration increased rapidly after 24 weeks and fluctuated between 220–280 mg/L after 43 weeks. The pH values were below 7 during the first 8 weeks and after that leachates appeared to be alkaline. Carbon dioxide was the main composition in landfill gas and its concentration remained at a high level through the whole stabilization process. The average contents of carbon dioxide, oxygen, and methane varied between 19 vol.%–28 vol.%, 2 vol.%–8 vol.%, and 5 vol.%–13 vol.%, respectively. A relative equilibrium was reached after 48 weeks. The highest temperature in the landfill chamber could amount to 75.8 degrees centigrade.     

Excavation and characterization of refuse in closed landfill

Refuse in landfills becomes stabilized as organic matter in refuse degrades and soluble inorganic substances dissolve during their longterm stabilization process.In this paper,this process is also referred to as mineralization process and the resultant stabilized refuse referred to as aged refuse.Aged refuse contains a wide spectrum and huge quantity of microorganisms with strong decomposition capability for refractory organic matter present in some wastewater such as leachate.In this study,aged refuse excavated from 2 to 10 years old closed landfill compartments in Shanghai Refuse Landfill is characterized in terms of particulate distribution by screening,total nitrogen,total phosphorus,biodegradable matter.The approaches for redevelopment of both land and aged refuse in the stabilized landfills are proposed.     

Three-stage aged refuse biofilter for the treatment of landfill leachate

A field-scale aged refuse(AR) biofilter constructed in Shanghai Refuse Landfill,containing about 7000 m3 aged refuse inside,was evaluated for its performance in the treatment of landfill leachate. This AR biofilter can be divided into three stages and can manage 50 m3 landfill leachate per day. The physical,chemical,and biological characteristics of AR were analyzed for evaluating the AR biofilter as leachate treatment host. The results revealed that over 87.8%-96.2% of COD and 96.9%-99.4% of ammonia nitrog...     

Comparison between controlled landfill reactor and conditioned landfill bioreactor

Bioreactor landfills allow a more active landfill management that recognizes the biological, chemical and physical processes involved in a landfill environment. The laboratory-scale simulators of landfill reactors treating municipal solid wastes were studied, the effect of solid waste size, leachate recirculation, nutrient balance, pH value, moisture content and temperature on the rate of municipal solid waste (MSW) biodegradation were determined, and it indicated the optimum pH value, moisture content and temperature decomposing MSW. The results of waste biodegradation were compared with that of the leachate-recirculated landfill simulator and conservative sanitary landfill simulator. In the control experiment the antitheses of a decreasing trend of the organic load, measured as biological oxygen demand and chemical oxygen demand, was shown. An obvious enhancement of effective disposal from conservative sanitary landfill (CSL) simulator, to the leachate-recirculated landfill (LRL) simulator and to the conditioned bioreactor landfill(CBL) simulator would be noted, through displaying the compared results of solid waste settlement, heavy, metal concentration in leachate, methane production rate, biogas composition, BOD and COD as well as their ratio.     

Enrichments of methanotrophic–heterotrophic cultures with high poly-β-hydroxybutyrate (PHB) accumulation capacities

Methanotrophic–heterotrophic communities were selectively enriched from sewage sludge to obtain a mixed culture with high levels of poly-β-hydroxybutyrate(PHB)accumulation capacity from methane.Methane was used as the carbon source,N_2as sole nitrogen source,and oxygen and Cu content were varied.Copper proved essential for PHB synthesis.All cultures enriched with Cu could accumulate high content of PHB(43.2%–45.9%),while only small amounts of PHB were accumulated by cultures enriched without Cu(11.9%–17.5%).Batch assays revealed that communities grown with Cu and a higher O_2content synthesized more PHB,which had a wider optimal CH_4:O_2range and produced a high PHB content(48.7%)even though in the presence of N_2.In all methanotrophic–heterotrophic communities,both methanotrophic and heterotrophic populations showed the ability to accumulate PHB.Although methane was added as the sole carbon source,heterotrophs dominated with abundances between 77.2%and 85.6%.All methanotrophs detected belonged to type II genera,which formed stable communities with heterotrophs of different PHB production capacities.     

甲烷氧化菌在填埋场覆盖层中的工程应用(Ⅱ):基质选择性及长效性

针对已筛选获得的甲烷氧化混合菌,选取填埋场覆盖土(LCS)、矿化垃圾(AR)和塘渣(TZ)3种填埋场周边易得的材料为供试生存基质,从基质选择性及长效性角度进行了甲烷减排应用条件的探究及使用效能评估.结果表明,在TZ、LCS、AR、TZ-AR和LCS-AR这5种生存基质中,TZ-AR最适合甲烷氧化混合菌的生长,且TZ与AR的复配比例以5∶5为最佳.甲烷氧化混合菌在TZ-AR的粒径≤4 mm和含水率为20%时具有最高甲烷氧化能力.一次性接种甲烷氧化混合菌在静态体系中的最佳使用有效期为31 d.其在接种量为0.08、0.16、0.20 m L·g-1和0.25m L·g-1时甲烷氧化速率无明显差异,从工程应用角度而言,8%的接种量为最佳.     

生活垃圾填埋场甲烷自然减排的新途径:厌氧与好氧的共氧化作用

采用暴雨过后垃圾填埋表层30～60 cm的覆土、表层1.5 m以下的垃圾,以及刚刚开挖出来的9年矿化垃圾进行硫酸盐还原菌阳性反应试验,结果表明生活垃圾填埋体不同填埋层都存在不同数量级的硫酸盐还原菌,且底层矿化垃圾中的硫酸盐还原菌的数量最多,表层覆土中最少.颗粒大小比例为50%:50%的垃圾样品表现出最佳的甲烷好氧与厌氧...     

硝酸盐对矿化垃圾中兼/厌氧甲烷氧化的影响

设计全因子实验研究了NO-3-N对填埋10~12 a的矿化垃圾中兼/厌氧甲烷氧化作用的影响.结果表明,兼/厌氧条件下,NO-3-N能促进矿化垃圾中CH4的去除.初始CH4和NO-3-N对CH4去除和N2产生有明显影响,且两者具有交互作用(P<0.05).CH4去除量随着初始CH4体积分数的增加而增加,添加一定含量的NO-3-N能促进CH4去除,同时通入一定体积分数CH4可以明显促进反硝化作用,说明矿化垃圾中NO-3-N还原能与兼/厌氧甲烷氧化耦合.本实验条件下,初始CH4体积分数为30%,NO-3-N含量为110 mg·kg-1时耦合效应较好.     

Interaction and independence on methane oxidation of landfill cover soil among three impact factors: water, oxygen and ammonium

To understand the influence patterns and interactions of three important environmental factors, i.e. soil water content, oxygen concentration, and ammonium addition, on methane oxidation, the soils from landfill cover layers were incubated under full factorial parameter settings. In addition to the methane oxidation rate, the quantities and community structures of methanotrophs were analyzed to determine the methane oxidation capacity of the soils. Canonical correspondence analysis was utilized to distinguish the important impact factors. Water content was found to be the most important factor influencing the methane oxidation rate and Type II methanotrophs, and the optimum value was 15% (w/w), which induced methane oxidation rates 10- and 6- times greater than those observed at 5% (w/w) and 20% (w/w), respectively. Ambient oxygen conditions were more suitable for methane oxidation than 3% oxygen. The addition of 100 mg-N·kg_drysoil⁻¹ of ammonium induced different effects on methane oxidation capacity when conducted at low or high water content. With regard to the methanotrophs, Type II was sensitive to the changes of water content, while Type I was influenced by oxygen content. Furthermore, the methanotrophic acidophile, Verrucomicrobia, was detected in soils with a pH of 4.9, which extended their known living environments.     

氨氧化菌对填埋场CH4协同氧化及铵抑制作用的研究

借助乙炔(C₂H₂)抑制和添加外源铵盐,采用批式培养试验,在初始CH₄浓度为16%的条件下模拟填埋场高CH₄浓度环境,通过分析样品中氨氧化菌对CH₄氧化的贡献率及铵对CH₄氧化的抑制率,研究了填埋场覆盖土、矿化垃圾、砂土和黏土中氨氧化菌对CH₄协同氧化及铵抑制作用. 结果表明:4种供试样品中氨氧化菌对CH₄氧化的贡献率在5.64%~16.24%之间,次序为砂土＜黏土＜覆盖土＜矿化垃圾,覆盖土中的贡献率为14.90%,比矿化垃圾低8.25%,填埋场样品(矿化垃圾和覆盖土)是一般土壤(砂土和黏土)的1.8~10.9倍. 铵对CH₄氧化过程的抑制率在11.90%~24.84%之间,次序为砂土＜黏土＜覆盖土＜矿化垃圾,覆盖土中为23.21%,比矿化垃圾低6.56%,填埋场样品是一般土壤的0.9~2.1倍. 填埋场样品中氨氧化菌对CH₄氧化的贡献率及铵对CH₄氧化的抑制率明显高于一般土壤.      

硝酸盐和甲烷对覆土中苯系物厌氧氧化的影响

苯系物是填埋场填埋气中恶臭有机气体的重要成分之一,填埋气中的CH4则是重要的温室气体.填埋覆土层中的微生物可以氧化CH4和苯系物,因此,强化微生物的氧化效能有助于削减和控制填埋气的污染.电子受体还原可耦合甲烷和某些有机物的厌氧氧化,从而去除甲烷和有机物.鉴此,本研究通过静态培养试验,分析了电子受体SO2-4共存条件下,NO-3和CH4共存对覆土中苯系物厌氧降解的影响.结果表明不外加NO-3时,苯系物抑制CH4的降解,加入NO-3后,苯系物共存反而有利于CH4的去除;单独添加NO-3或CH4都能促进填埋覆土中苯系物的去除;而同时添加NO-3和CH4能更好地促进苯系物的去除,甲苯、二甲苯和异丙苯的去除率最高可达65%、88%和82%,远高于不添加NO-3和CH4对照处理的53%、76%和31%;NO-3还原与CH4厌氧氧化耦合过程能同步促进苯系物的厌氧氧化.