Landfill methane oxidation in soil and bio-based cover systems: a review

Mitigation of landfill gases has gained the utmost importance in recent years due to the increase in methane (CH₄) emissions from landfills worldwide. This, in turn, can contribute to global warming and climatic changes. The concept of microbially mediated methane oxidation in landfill covers by using methanotrophic microorganisms has been widely adopted as a method to counter the rise in methane emissions. Traditionally, landfill soil covers were used to achieve methane oxidation, thereby reducing methane emissions. Meanwhile, the continual rise of CH₄ emissions from landfills and the significant need to and importance of developing a better technology has led researchers to explore different methods to enhance microbial methane oxidation by using organic rich materials such as compost in landfill covers. The development and field application of such bio-based systems, explored by various researches worldwide, eventually led to more widely accepted and better performing cover systems capable of reducing CH₄ emissions from landfills. However, the long-term performance of bio-based cover systems were found to be negatively affected by factors such as the material’s ability to self-degrade, causing CH₄ to be generated rather than oxidized as well as the greater potential for forming pore-clogging exopolymeric substances. In order to design an effective cover system for landfills, it is essential to have a thorough understanding of the concepts incorporated into methodologies currently in favor along with their pros and cons. This review summarizes previous laboratory and field-scale studies conducted on various soil and bio-based cover systems, along with the modeling mechanisms adopted for quantifying CH₄ oxidation rates. Finally, several issues and challenges in developing effective and economical soil and bio-based cover systems are presented.     

Methanogenic interactions in model landfill co-cultures with paper as the carbon source

In a simple laboratory model system containing co-cultures of cellulolytic bacteria from landfills and Methanobrevibacter smithii , methanogenesis was optimal at 37°C. Although paper solubilization was greatest in cultures containing Eubacterium strain LF 9, methanogenesis was greatest in cultures containing Eubacterium strain LF 11. Coating filter paper with printer's ink reduced methanogenesis. Cultures grown on newspaper produced less methane than those grown on filter paper. However, the rates of methane production from these substrates were similar when calculated in relation to microbial biomass present.     

增温对冻土区泥炭沼泽土壤孔隙水甲烷关联微生物和溶解性有机碳的影响

多年冻土区泥炭沼泽土壤孔隙水甲烷关联微生物及底物的研究有助于深入理解气候变化背景下寒区湿地生态系统甲烷循环过程。选取大兴安岭连续多年冻土区柴桦-泥炭藓和狭叶杜香-泥炭藓两种典型植被群落泥炭沼泽,设置开顶箱(Open Top Chamber,OTC)增温实验。于生长季(6月、7月、8月和9月)采集土壤孔隙水样品,对比分析OTC内外土壤孔隙水中产甲烷菌数量、甲烷氧化菌数量及溶解性有机碳(Dissolved Organic Carbon,DOC)浓度的动态变化特征,并探究土壤孔隙水甲烷关联微生物与DOC浓度的关系。结果表明:增温提高了生长季大兴安岭多年冻土区土壤孔隙水中产甲烷菌数量和DOC浓度,而对甲烷氧化菌数量的影响因月份而异。生长季柴桦-泥炭藓和狭叶杜香-泥炭藓泥炭沼泽土壤孔隙水中产甲烷菌数量的平均增加幅度分别为54.52%和44.97%,DOC浓度的平均增加幅度分别为34.16%和28.33%。增温使得生长季柴桦-泥炭藓和狭叶杜香-泥炭藓泥炭沼泽土壤孔隙水中甲烷氧化菌平均数量分别降低了46.20%和31.42%。一元线性回归分析结果表明,土壤孔隙水中DOC浓度可分别解释柴桦-泥炭藓和狭叶杜香-泥炭藓泥炭沼泽土壤孔隙水中产甲烷菌数量变化的29.00%和24.10%(P<0.01),而对两种植被群落下土壤孔隙水中甲烷氧化菌数量的影响并不显著(P>0.05)。     

Elimination of methane generated from landfills by biofiltration: a review

The production of biogas in landfills, its composition and the problems resulting from its generation are all reviewed. Biofiltration is a promising option for the control of emissions to atmosphere of the methane contained in biogas issued from the smaller and/or older landfills. A detailed review of the methane biofiltration literature is presented. The microorganisms, mainly the methanotrophs, involved in the methane biodegradation process, and their needs in terms of oxygen and carbon dioxide utilization, are described. Moreover, the influence of nutrients such as copper, nitrogen and phosphorus, and the process operating conditions such as temperature, pH and moisture content of the biofilter bed, are also presented. Finally, the performance of various filter beds, in terms of their elimination capacities, is presented for laboratory scale biofilters and landfill covers.     

垃圾填埋场N2O排放通量及测定方法研究进展

氧化亚氮(N₂O)是第三大温室气体和最主要的臭氧层破坏气体.填埋是目前城市生活垃圾处理处置的主要方式,而垃圾填埋场是N₂O的排放源之一.实验室研究和现场测定均表明,生活垃圾填埋场可以有高的N₂O释放通量,但不同填埋场测定数据差异很大.目前,对生活垃圾填埋场N₂O排放量的原位准确测定以及排放机理和重要性的认识仍有很多不足.本文概述了生活垃圾填埋场N₂O排放研究现状,从垃圾堆体和覆土层两部分探讨了传统厌氧卫生填埋场的N₂O产生和排放机理,并就此对新型脱氮型生物反应器填埋场做了相应探讨.最后,就静态箱法、涡度相关法等N₂O通量测定方法在填埋场的适用性进行了讨论,并展望了填埋场N₂O排放的研究方向.     

Extraction of methane-oxidizing bacteria from soil particles

Abstract: We present a method for extraction of active methane (CH₄)-oxidizing bacteria from soil samples. The method is based on physical dispersion of bacteria from the soil particles followed by separation of bacteria and soil particles by floatation in the density media Nycodenz or Percoll. Separation on Nycodenz produced very pure bacterial suspensions while separation on Percoll produced rather impure suspensions. However, more than 60% of the methane-oxidizing activity was irreversibly inhibited in the procedure using Nycodenz compared to less than 10% irreversible inhibition when Percoll was employed. The bacterial suspensions extracted from soil can be used to study the physiology and ecology of soil bacteria that oxidize methane at atmospheric concentrations. Our data indicated that these bacteria are extremely difficult to dislodge from particles compared to the majority of bacteria in soil. Tentatively, we interpret the strong attachment to long residence time (i.e. slow turnover) of the methane-oxidizing bacteria. A slow turnover/growth rate would explain why soil disturbances, like cultivation, have a long lasting effect on the oxidation of atmospheric methane in soil.     

Microbial transformation of chitin in soil under anaerobic conditions

Anaerobic chitinolytic complex was studied in three soil types: chernozem, gray forest soil, and chestnut soil. The abundance and biomass of anaerobic chitinolytic microbial complex of fungi, bacteria, and actinomycetes were evaluated by luminescent microscopy. The dynamics of methane emission from soil during chitinolytic succession was studied by gas chromatography. All three studied microbial groups proved to participate in chitin transformation in soil under anaerobic conditions. The highest biomass growth was observed among prokaryotes, particularly actinomycetes, whose biomass doubled. The increase in methane emission during chitinolytic succession was most pronounced in soils with low organic matter content.     

Characterization of methanotrophic bacterial populations in soils showing atmospheric methane uptake.

The global methane cycle includes both terrestrial and atmospheric processes and may contribute to feedback regulation of the climate. Most oxic soils are a net sink for methane, and these soils consume approximately 20 to 60 Tg of methane per year. The soil sink for atmospheric methane is microbially mediated and sensitive to disturbance. A decrease in the capacity of this sink may have contributed to the approximately 1%. year(-1) increase in the atmospheric methane level in this century. The organisms responsible for methane uptake by soils (the atmospheric methane sink) are not known, and factors that influence the activity of these organisms are poorly understood. In this study the soil methane-oxidizing population was characterized by both labelling soil microbiota with (14)CH(4) and analyzing a total soil monooxygenase gene library. Comparative analyses of [(14)C]phospholipid ester-linked fatty acid profiles performed with representative methane-oxidizing bacteria revealed that the soil sink for atmospheric methane consists of an unknown group of methanotrophic bacteria that exhibit some similarity to type II methanotrophs. An analysis of monooxygenase gene libraries from the same soil samples indicated that an unknown group of bacteria belonging to the alpha subclass of the class Proteobacteria was present; these organisms were only distantly related to extant methane-oxidizing strains. Studies on factors that affect the activity, population dynamics, and contribution to global methane flux of "atmospheric methane oxidizers" should be greatly facilitated by use of biomarkers identified in this study.     

The biogeochemical controls of N2O production and emission in landfill cover soils: the role of methanotrophs in the nitrogen cycle

Emissions of N2O from cover soils of both abandoned (> 30 years) and active landfills greatly exceed the maximum fluxes previously reported for tropical soils, suggesting high microbial activities for N2O production. Low soil matrix potentials (<-0.7 MPa) indicate that nitrification was the most likely mechanism of N2O formation during most of the time of sampling. Soil moisture had a strong influence on N2O emissions. The production of N2O was stimulated by as much as 20 times during laboratory incubations, when moisture was increased from -2.0 MPa to -0.6 MPa. Additional evidence from incubation experiments and delta13C analyses of fatty acids (18:1) diagnostic of methanotrophs suggests that N2O is formed in these soils by nitrification via methanotrophic bacteria. In a NH3(g)-amended landfill soil, the rate of N2O production was significantly increased when incubated with 100 ppmv methane compared with 1.8 ppmv (atmospheric) methane. Preincubation of a landfill soil with 1% CH4 for 2 weeks resulted in higher rates of N2O production when subsequently amended with NH3(g) relative to a control soil preincubated without CH4. At one location, at the soil depth (9-16 cm) of maximum methane consumption and N2O production, we observe elevated concentrations of organic carbon and nitrogen and distinct minima in delta15N (+1.0%) and delta13C (-33.8%) values for organic nitrogen and organic carbon respectively. A delta13C value of -39.3% was measured for 18:1 carbon fatty acids in this soil, diagnostic of type II methanotrophs. The low delta15N value for organic nitrogen is consistent with N2 fixation by type II methanotrophs. These observations all point to a methanotrophic origin for the organic matter at this depth. The results of this study corroborate previous reports of methanotrophic nitrification and N2O formation in aqueous and soil environments and suggest a predominance of type II rather than type I or type X methanotrophs in this landfill soil.     

Responses of atmospheric methane consumption by soils to global climate change

Soils consume about 40 Tg methane from the atmosphere annually. Thus, soils contribute significantly to the atmospheric methane budget. However, responses of atmospheric methane consumption to climate change are uncertain. Predicting these responses requires an understanding of the effect on methane consumption of specific variables (temperature and soil water content) as well as interactions among parameters (methane, ammonium, water content). Key considerations involve the limitations of diffusive transport and controls of methane diffusivity; limitation of methanotrophic activity by water stress; relatively slow growth rates of methane-oxidizing bacteria on atmospheric methane; ammonium toxicity. Interactions among these parameters may be particularly important, and lead to responses contrary to those predicted from changes in temperature and water content alone. Results from a number of analyses indicate that atmospheric methane consumption is especially sensitive to anthropogenic disturbances, which typically decrease activity. Continued increases in wet and dry ammonium deposition are likely to exacerbate inhibition resulting from changes in land use. Changes in hydrological regimes could further decrease activity if dry periods increase water stress at soil depths currently colonized by methanotrophs. Future trends in the soil methane sink are likely to lead to enhanced accumulation of atmospheric methane.     

稻田土壤亚硝酸盐型甲烷厌氧氧化菌群落结构的时空特征

稻田是温室气体甲烷的重要排放源之一,对全球气候变化具有重要影响.由隶属于NC10门的Candidatus Methylomirabilis oxyfera(M.oxyfera)-like细菌介导的亚硝酸盐型甲烷厌氧氧化是控制稻田甲烷排放的新途径.目前,有关此类微生物群落在稻田土壤中的时空分布特征及其环境影响因素尚不明确...     

Physiological Studies of Methane- and Methanol-Oxidizing Bacteria: Immunological Comparison of a Primary Alcohol Dehydrogenase from Methylococcus capsulatus and Pseudomonas sp. M27

A primary alcohol dehydrogenase was purified from cell extracts of two apparently unrelated microorganisms, namely, Pseudomonas sp. M27 and Methylococcus capsulatus. Rabbit antiserum prepared against the purified enzyme from M. capsulatus revealed distinctive antigenic determinants by quantitative and gel precipitin reactions. Rabbit antiserum to M27 enzyme detected both distinctive and shared antigenic determinants. Certain methane- and methanol-oxidizing bacteria were grouped on the basis of serological cross-reacting enzyme specificities.     

Effect of various anionic species on net methane production in flooded rice soils

In a laboratory incubation study, effect of various anions on net methane production in two rice soils (alluvial and acid sulphate) under flooded conditions was examined. Methane production was considerable in alluvial soil and almost negligible in acid sulphate soil, albeit with a higher density of viable methanogens, during 30-day incubation without salts. Sodium salts of hydroxide and phosphate further stimulated methane production in alluvial soil and marginally in acid sulphate soil. But, addition of sodium molybdate, a selective inhibitor of sulphate-reducing bacteria, increased the production of methane in acid sulphate soil. In contrast, nitrite, nitrate, sulphite and sulphate suppressed the production of methane in both soils. Acetate served as an excellent substrate for methanogenesis in alluvial soil, but not in acid sulphate soil. Succinate and citrate also stimulated methane production especially in alluvial soil, but after a longer lag. In acid sulphate soil, most of the added carbon in the form of sodium salts of carboxylic acids was converted to CO2 and not methane, which is consistent with their preferential use by the sulphate-reducing bacteria. In general, none of the amendments could increase production of methane in acid sulphate soil to the same level as in alluvial soil.     

Effect of ducks on CH4 emission from paddy soils and its mechanism research in the rice-duck ecosystem

The effect of ducks on CH₄ emission from paddy soils and its mechanism were probed in order to decide the optimum number of ducks in the rice-duck ecosystem. Methane emission fluxes from paddy soils were measured by the static box technique. The correlations between methane emission and soil physical and chemical characteristics were also analyzed. The results showed that significant differences (p < 0.01) existed in the dissolved oxygen content of water body in the treatment fields, and the more the ducks, the higher the dissolved oxygen content. Secondly, the soil redox matter content and methanogenic bacteria population of the rice-duck ecosystem reduced more sharply than those of the no-duck rice farming, resulting in a lower methane production. Thirdly, the amount of methane emission differed between the treatments—the more the ducks, the less the methane emission. Other related analyses showed that the negative correlation was significant (p < 0.001) between the methane emission flux and dissolved oxygen content of water body. However, CH4 emission flux had significantly positive correlation (p < 0.01) with the soil redox matter content and rice field methanogenic population.     

典型草地土壤好氧甲烷氧化的微生物生态过程

【目的】针对我国甘肃三个典型生态区草地土壤(玛曲MQ、临泽LZ和环县HX),研究其甲烷氧化潜力、甲烷氧化菌(methane-oxidizingbacteria,MOB)丰度及可能存在的群落分异规律。【方法】通过原位分析、室内高浓度甲烷模拟培养三种典型土壤及实时荧光定量、高通量测序的方法研究甲烷氧化菌标靶基因pmoA序列的组成及其丰度变化规律。【结果】三种典型草地土壤的原位甲烷氧化菌的丰度存在显著差异,表现为MQ>HX>LZ,其数量范围为为0.18–6.86×10^7g/d.w.s.;甲烷氧化潜力也表现出类似规律,其通量为109–169mg/(m^2·h);甲烷氧化潜力与原位土壤中甲烷氧化菌丰度有正相关。三种草地土壤甲烷氧化菌存在明显的空间异质性,采用高通量测序的方法,发现三种草地原位土壤中的优势类群为USCγ(Upland Soil Cluster gamma,USCγ);然而,室内高浓度甲烷氧化过程中,传统的甲烷氧化菌均发生明显增加,MQ土壤中TypeⅡ的Methylocystis为优势类群,而LZ和HX土壤的优势类群均为TypeⅠ型Methylosarcina。【结论】这些研究结果表明,我国甘肃典型草地土壤中也存在难培养的大气甲烷氧化菌和经典的可培养甲烷氧化菌,这些微生物极可能氧化极低浓度的大气甲烷,也可能利用闭蓄于土壤中的高浓度甲烷生长。未来应采用先进技术原位观测大气甲烷氧化过程并分离相应微生物类群,研究草地土壤甲烷氧化菌地理分异规律及其环境驱动机制。     

Studies on methanol-oxidizing bacteria

Three methanol-oxidizing bacteria were isolated by enrichment culture technique from soil. Two of them werePseudomonas spp. The third one was obligate methylotroph. Their growth characteristics have been described. Cell-suspension experiments withPseudomonas RJ₁ suggest that methanol-, formaldehyde-, and formate-oxidizing enzymes were present. Formate-oxidizing enzyme was detected only fromPseudomonas RJ₁. It had a pH optimum of 7.0 and required nicotinamide adenine dinucleotide (NAD) for activity. Cyanide at 1 × 10⁻⁵ m concentration inhibited the enzyme activity completely.     

湿地松和马尾松人工林土壤甲烷代谢微生物群落的结构特征

土壤甲烷代谢微生物影响甲烷的产生和氧化,然而关于林型对土壤中甲烷代谢微生物群落结构影响的研究较少。采用基因芯片GeoChip 3.0研究了湿地松人工林和马尾松人工林土壤甲烷代谢微生物群落结构特征。结果如下,(1)两种林型的甲烷代谢微生物群落结构存在极显著差异(P=0.008),林型能解释其34.9%的变异;(2)产甲烷菌(包含甲基辅酶M还原α亚基基因mcrA的微生物)的优势菌群发生了变化,湿地松人工林的的优势菌为Methanocorpusculum labreanum Z,马尾松人工林的优势菌群除Methanocorpusculum labreanum Z外,还包括产甲烷古菌和Methanosarcina mazei Gol;(3)甲烷营养菌(包含甲烷单加氧酶基因pmoA基因的微生物)的优势菌为Ⅱ型,有3种不可培养细菌只在湿地松人工林检测到,在马尾松人工林中未检测到;(4)mcrA基因丰度或同源基因数量与土壤容重正相关,与土壤粘粒含量呈显著负相关;pmoA基因信号强度或多样性指数与土壤全碳含量、全磷含量和速效氮含量显著正相关。总之,相比本地种马尾松人工林,引进种湿地松人工林的土壤甲烷代谢微生物群落结构发生了显著变化。     

Release of fossil methane from mineral soil particles, and its implication for estimation of methane oxidation in a mineral subsoil

In a preliminary experiment we found that methane evolved from a sandy subsoil during aerobic incubation of shaken soil slurries. In the study presented here the methane was found to be released from the sand particles by mechanical weathering, caused by the grinding effect of the shaking. Large amounts of gas (about 0.5 ml gas g^–1 soil) were extracted by intense grinding of the soil in gas tight serum vials. Methane was the main hydrocarbon in the emitted gas, but also a considerable amount of ethane was present, as well as minor amounts of heavier hydrocarbons (up to C₆). The ¹³C-values of the emitted methane and ethane were –33 and –29 , respectively. Together these results demonstrate a thermogenic origin of the gas. This paper also reports the results of an incubation experiment where possible methane oxidation was looked for. If a possible release of methane is not accounted for, methane oxidation may be overlooked, as illustrated in this paper. Methane consumption was detected only in soil from 40 cm, in contrast to soil sampled at 100 cm and deeper where a slight production was measured. When methane oxidation was inhibited by dimethyl-ether, a significant release of methane was seen. The release was probably caused by chemical weathering. When this methane release was taken into account, methane oxidation was found to be present at all measured depths (40 to 200 cm). Fertilization with urea inhibited the methane oxidation at 40 cm but not at deeper layers. It is hypothesized that ammonia oxidizing bacteria were the main methane oxidizers in this mineral subsoil (deeper than 1 m), and that oxidation of methane might be a survival mechanism for ammonia oxidizers in ammonia limited environments.     

Applications of a colorimetric plate assay for soluble methane monooxygenase activity.

A straightforward method is described for screening methanotrophic colonies for soluble methane monooxygenase (sMMO) activity on solid media. Such activity results in the development of a colored complex between 1-naphthol, which is formed when sMMO reacts with naphthalene, and o-dianisidine (tetrazotized). Methanotrophic colonies expressing sMMO turned deep purple when exposed successively to naphthalene and o-dianisidine. The method was evaluated within the contexts of two potential applications. The first was for the enumeration of Methylosinus trichosporium OB3b in a methane-amended, unsaturated soil column dedicated to vinyl chloride treatment. The second application was for the isolation and enumeration of sMMO-bearing methanotrophs from sanitary landfill soils. The technique was effective in both applications.     

Radioactive fingerprinting of microorganisms that oxidize atmospheric methane in different soils.

Microorganisms that oxidize atmospheric methane in soils were characterized by radioactive labelling with (14)CH(4) followed by analysis of radiolabelled phospholipid ester-linked fatty acids ((14)C-PLFAs). The radioactive fingerprinting technique was used to compare active methanotrophs in soil samples from Greenland, Denmark, the United States, and Brazil. The (14)C-PLFA fingerprints indicated that closely related methanotrophic bacteria were responsible for the oxidation of atmospheric methane in the soils. Significant amounts of labelled PLFAs produced by the unknown soil methanotrophs coeluted with a group of fatty acids that included i17:0, a17:0, and 17:1omega8c (up to 9.0% of the total (14)C-PLFAs). These PLFAs are not known to be significant constituents of methanotrophic bacteria. The major PLFAs of the soil methanotrophs (73.5 to 89.0% of the total PLFAs) coeluted with 18:1 and 18:0 fatty acids (e.g., 18:1omega9, 18:1omega7, and 18:0). The (14)C-PLFAs fingerprints of the soil methanotrophs that oxidized atmospheric methane did not change after long-term methane enrichment at 170 ppm CH(4). The (14)C-PLFA fingerprints of the soil methanotrophs were different from the PLFA profiles of type I and type II methanotrophic bacteria described previously. Some similarity at the PLFA level was observed between the unknown soil methanotrophs and the PLFA phenotype of the type II methanotrophs. Methanotrophs in Arctic, temperate, and tropical regions assimilated between 20 and 54% of the atmospheric methane that was metabolized. The lowest relative assimilation (percent) was observed for methanotrophs in agricultural soil, whereas the highest assimilation was observed for methanotrophs in rain forest soil. The results suggest that methanotrophs with relatively high carbon conversion efficiencies and very similar PLFA compositions dominate atmospheric methane metabolism in different soils. The characteristics of the methane metabolism and the (14)C-PLFA fingerprints excluded any significant role of autotrophic ammonia oxidizers in the metabolism of atmospheric methane.