Methane oxidation in landfill cover soil

Methane oxidation in the cover soil of the Khmet'evo municipal landfill in Moscow oblast was investigated. Methane emission from the experimental parcel of the site was highly inhomogeneous. At a depth of 45-60 cm, the pore gas mainly consisted of CH4 (60-70%) and CO2 (30-40%). In the upper horizons of the cover soil, the concentration of these gases sharply decreased. Techniques for estimation of the methane-oxidizing activity in the cover soil of the landfill were tested. The rate of methane oxidation in the soil, the factor limiting methane emission from the surface of the site, correlated with the cell number of culturable methanotrophic bacteria. The method of indirect immunofluorescence revealed ten known species of methanotrophic bacteria in enrichment cultures obtained from samples of the cover soil. Our results also indicate the presence of unknown psychrotolerant methanotrophs that are active at the low temperatures characteristic of Moscow oblast.     

填埋覆土甲烷氧化微生物及甲烷氧化作用机理研究进展

甲烷是一种长期存在于大气中的温室气体,它对温室效应的贡献率是二氧化碳的26倍.生活垃圾填埋场是大气甲烷的主要产生源之一,由其产生的甲烷约占全球甲烷排放总量的1.5%～15%.甲烷氧化微生物在调节全球甲烷平衡中起着重要作用.垃圾填埋场覆土具有相当强的甲烷氧化能力.填埋覆土甲烷氧化菌及其氧化作用机理的研究,已成为环境微生物学研究领域的热点之一.本文对生活垃圾填埋场填埋覆土中甲烷氧化微生物、甲烷氧化机理及动力学机制、甲烷与微量填埋气体的共氧化机制以及影响甲烷氧化的环境因子研究的最新进展进行综述,并对生活垃圾填埋场甲烷氧化微生物的研究进行展望.     

Activity of a Methanotrophic Consortium Isolated from Landfill Cover Soil: Response to Temperature,pH, CO2, and Porous Adsorbent

A robust, naturally evolving methanotrophic community in landfill cover soil (LFCS) can be the simplest way to mitigate landfill methane emission. In this study, bacterial community composition in LFCS and methane oxidation potential of enriched methanotrophic consortium, in comparison to that of axenic Methylosinus sporium, was investigated. Growth and methane oxidation of the consortium was studied in liquid phase batch experiments under varying temperature (20–40°C), pH (5–10), headspace CO2, and in presence of porous adsorbent (1.3 cm³ sponge cubes). The 16S rRNA gene analysis revealed presence of both type-I and type-II methanotrophs along with few obligate methylotroph in LFCS. Though the optimal growth condition of the consortium was at 30°C and pH 7, it was more resilient in comparison to M. sporium. With increasing availability of porous adsorbent, methane consumption by the consortium was significantly improved (p < 0.001) reaching a maximum specific methane oxidation rate of 11.4 μmol mg^?1 biomass h^?1. Thus, inducing naturally thriving methanotrophs in LFCS is a better alternative to axenic methanotrophic culture in methane emission management.     

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.     

Quantification of methane oxidation in the rhizosphere of emergent aquatic macrophytes: defining upper limits

Rates of rhizospheric methane oxidation were evaluated by aerobic incubations of subcores collected in flooded anoxic soils populated by emergent macrophytes, by greenhouse whole plant incubations, and by CH₄ stable isotopic analysis. Subcore incubations defined upper limits for rhizospheric methane oxidation on an areal basis which were equal to or greater than emission rates. These rates are considered upper limits because O₂ did not limit CH₄ uptake as is likely to occur in situ. The ratio of maximum potential methane oxidation (MO) to methane emission (ME) ranged from 0.7 to 1.9 in Louisiana rice (Oryza sativa), from 1.0 to 4.0 in a N. Florida Sagittaria lancifolia marsh, and from 5.6 to 51 in Everglades Typha domingensis and Cladium jamaicense areas. Methane oxidation/methane emission ratios determined in whole plant incubations of Sagittaria lancifolia under oxic and anoxic conditions ranged from 0.5 to 1.6. Methane oxidation activity associated with emergent aquatic macrophytes was found primarily in fine root material. A weak correlation was observed between live root biomass and CH₄ uptake in Typha. Rhizomes showed small or zero rates of methane uptake and no uptake was associated with plant stems. Methane stable isotope data from a S. lancifolia marsh were as follows: CH₄ emitted from plants: −61.6 ± 0.3%; CH₄ within stems: −42.0 ± 0.2%; CH₄ within sedimentary bubbles: −51.7 ± 0.3%). The ¹³C enrichment observed relative to emitted CH₄ could be due to preferential mobilization of CH₄ containing the lighter isotope and/or the action of methanotrophic bacteria.     

Microbiological and biogeochemical processes in a pockmark of the Gdansk depression,Baltic Sea

Comprehensive microbiological and biogeochemical investigation of a pockmark within one of the sites of gas-saturated sediments in the Gdansk depression, Baltic Sea was carried out during the 87th voyage of the Professor Shtokman research vessel. Methane content in the near-bottom water and in the underlying sediments indicates stable methane flow from the sediment into the water. In the 10-m water layer above the pockmark, apart from methane anomalies, elevated numbers of microorganisms and enhanced rates of dark CO₂ fixation (up to 1.15 µmol C/(l day)) and methane oxidation (up to 2.14 nmol CH₄/(l day)) were revealed. Lightened isotopic composition of suspended organic matter also indicates high activity of the near-bottom microbial community. Compared to the background stations, methane content in pockmark sediments increased sharply from the surface to 40–60 ml/dm³ in the 20–30 cm horizon. High rates of bacterial sulfate reduction (SR) were detected throughout the core (0–40 cm); the maximum of 74 µmol S/(dm³ day) was located in subsurface horizons (15–20 cm). The highest rates of anaerobic methane oxidation (AMO), up to 80 µmol/dm³ day), were detected in the same horizon. Good coincidence of the AMO and SR profiles with stoichiometry close to 1: 1 is evidence in favor of a close relation between these processes performed by a consortium of methanotrophic archaea and sulfate-reducing bacteria. Methane isotopic composition in subsurface sediments of the pockmark (from ?53.0 to ?56.5‰) does not rule out the presence of methane other than the biogenic methane from the deep horizons of the sedimentary cover.     

The processes of methane formation and oxidation in the soils of the Russian arctic tundra

Methane emission from the following types of tundra soils was studied: coarse humic gleyey loamy cryo soil, peaty gley soil, and peaty gleyey midloamy cryo soil of the arctic tundra. All the soils studied were found to be potential sources of atmospheric methane. The highest values of methane emission were recorded in August at a soil temperature of 8-10 degrees C. Flooded parcels were the sources of atmospheric methane throughout the observation period. The rates of methane production and oxidation in tundra soils of various types at 5 and 15 degrees C were studied by the radioisotope method. Methane oxidation was found to occur in bog water, in the green part of peat moss, and in all the soil horizons studied. Methane formation was recorded in the horizons of peat, in clay with plant roots, and in peaty moss dust of the bogey parcels. At both temperatures, the methane oxidation rate exceeded the rate of methane formation in all the horizons of the mossy-lichen tundra and of the bumpy sinkhole complex. Methanogenesis prevailed only in a sedge-peat moss bog at 15 degrees C. Enrichment bacterial cultures oxidizing methane at 5 and 15 degrees C were obtained. Different types of methanotrophic bacteria were shown to be responsible for methane oxidation under these conditions. A representative of type I methylotrophs oxidized methane at 5 degrees C, and Methylocella tundrae, a psychroactive representative of an acidophilic methanotrophic genus Methylocella, at 15 degrees C.     

The processes of methane production and oxidation in the soils of the Russian Arctic tundra

Methane emission from the following types of tundra soils was studied: coarse humic gleyey loamy cryo soil, peaty gleyey soil, and peaty gleyey midloamy cryo soil of the arctic tundra. All the soils studied were found to be potential sources of atmospheric methane. The highest values of methane emission were recorded in August at a soil temperature of 8–10°C. Flooded parcels were the sources of atmospheric methane throughout the observation period. The rates of methane production and oxidation in tundra soils of various types were studied by the radioisotope method at 5 and 15°C. Methane oxidation was found to occur in bog water, in the green part of peat moss, and in all the soil horizons studied. Methane production was recorded in the horizons of peat, in clay with plant roots, and in peaty moss dust of the bogey parcels. At both temperatures, the methane oxidation rate exceeded the rate of methane production in all the horizons of the mossy-lichen tundra and of the hillock tundra with flat-bottom depressions. Methanogenesis prevailed only in a sedge-peat moss bog at 15°C. Bacterial enrichment cultures oxidizing methane at 5 and 15°C were obtained. Different types of methanotrophic bacteria were shown to be responsible for methane oxidation under these conditions. A representative of type I methylotrophs oxidized methane at 5°C, and Methylocella tundrae, a psychroactive representative of an acidophilic methanotrophic genus Methylocella, at 15°C.__________Translated from Mikrobiologiya, Vol. 74, No. 2, 2005, pp. 261–270.Original Russian Text Copyright © 2005 by Berestovskaya, Rusanov, Vasileva, Pimenov.     

Effect of Afforestation and Reforestation of Pastures on the Activity and Population Dynamics of Methanotrophic Bacteria

We investigated the effect of afforestation and reforestation of pastures on methane oxidation and the methanotrophic communities in soils from three different New Zealand sites. Methane oxidation was measured in soils from two pine (Pinus radiata) forests and one shrubland (mainly Kunzea ericoides var. ericoides) and three adjacent permanent pastures. The methane oxidation rate was consistently higher in the pine forest or shrubland soils than in the adjacent pasture soils. A combination of phospholipid fatty acid (PLFA) and stable isotope probing (SIP) analyses of these soils revealed that different methanotrophic communities were active in soils under the different vegetations. The C₁₈ PLFAs (signature of type II methanotrophs) predominated under pine and shrublands, and C₁₆ PLFAs (type I methanotrophs) predominated under pastures. Analysis of the methanotrophs by molecular methods revealed further differences in methanotrophic community structure under the different vegetation types. Cloning and sequencing and terminal-restriction fragment length polymorphism analysis of the particulate methane oxygenase gene (pmoA) from different samples confirmed the PLFA-SIP results that methanotrophic bacteria related to type II methanotrophs were dominant in pine forest and shrubland, and type I methanotrophs (related to Methylococcus capsulatus) were dominant in all pasture soils. We report that afforestation and reforestation of pastures caused changes in methane oxidation by altering the community structure of methanotrophic bacteria in these soils.     

Molecular analysis of high-affinity methane-oxidizing enrichment cultures isolated from a forest biocenosis and agrocenoses

Methane oxidation by microorganisms inhabiting aerobic soils is a key process involved in the regulation of the concentration of this significant greenhouse gas in the atmosphere; however, the microorganisms responsible for this process remain unknown. Three stable methane-oxidizing cultures were isolated from samples of forest soils (FS) and agricultural soils (AS) of Moscow oblast, as well as from soil samples collected from a Belgian agrocenosis (BS). The obtained enrichment cultures exhibit a high affinity for methane; their k_m values range from 54.2 to 176.8 nM CH₄ and are comparable to those of aerobic soils. Analysis of the fragments of the ribosomal (16S rRNA) and functional (pmoA) genes of methanotrophs by PCR— DGGE and cloning demonstrated the presence of bacteria belonging to the genera Methylocystis in FS, Methylosinus in AS and BS, and Methylocella in BS. It was established that Methylocystis and Methylosinus detected in the enrichment cultures contain the genes encoding the synthesis of the active center of two membrane-bound particulate methane monooxygenases; it is likely that one of these genes (pmoA2) is responsible for the capacity of these microorganisms for oxidation of atmospheric methane.     

Methane Oxidation and the Competition for Oxygen in the Rice Rhizosphere

A mechanistic approach is presented to describe oxidation of the greenhouse gas methane in the rice rhizosphere of flooded paddies by obligate methanotrophic bacteria. In flooded rice paddies these methanotrophs compete for available O₂ with other types of bacteria. Soil incubation studies and most-probable-number (MPN) counts of oxygen consumers show that microbial oxygen consumption rates were dominated by heterotrophic and methanotrophic respiration. MPN counts of methanotrophs showed large spatial and temporal variability. The most abundant methanotrophs (a Methylocystis sp.) and heterotrophs (a Pseudomonas sp. and a Rhodococcus sp.) were isolated and characterized. Growth dynamics of these bacteria under carbon and oxygen limitations are presented. Theoretical calculations based on measured growth dynamics show that methanotrophs were only able to outcompete heterotrophs at low oxygen concentrations (frequently <5 μM). The oxygen concentration at which methanotrophs won the competition from heterotrophs did not depend on methane concentration, but it was highly affected by organic carbon concentrations in the paddy soil. Methane oxidation was severely inhibited at high acetate concentrations. This is in accordance with competition experiments between Pseudomonas spp. and Methylocystis spp. carried out at different oxygen and carbon concentrations. Likely, methane oxidation mainly occurs at microaerophilic and low-acetate conditions and thus not directly at the root surface. Acetate and oxygen concentrations in the rice rhizosphere are in the critical range for methane oxidation, and a high variability in methane oxidation rates is thus expected.     

Seasonal Dynamics of Atmospheric Methane Oxidation in Gray Forest Soils

Seasonal fluctuations in the methane fluxes in the soil–atmosphere system were determined for gray forest soils of Central Russia. Consumption of atmospheric methane was found to exceed methane emission in gray forest soils under forest and in the agrocenosis. The average annual rates of atmospheric methane consumption by the soil under forest and in the agrocenosis were 0.026 and 0.008 mg C-CH₄/(m² h), respectively. The annual rate of atmospheric methane oxidation in the gray forest soils of Moscow oblast was estimated to be 0.68 kton. Seasonal fluctuations in the methane oxidation activity were due to changes in the hydrothermal conditions and in the reserves of readily decomposable organic matter and mineral nitrogen, as well as to changes in the activity of methane oxidizers.     

Methane oxidation rates in forest soils and their controlling variables: a review and a case study in Korea

Methane is one of the strongest of the greenhouse gases, being 30-fold more radiatively active than carbon dioxide on a molar basis. In addition, its atmospheric concentrations have increased by 1% per year since the Industrial Revolution. As such, the dynamics of methane is of great importance for the prediction of global climatic changes caused by increasing concentrations of greenhouse gases in the atmosphere. One of the most important biological sinks for methane is forest soils, where methanotrophic bacteria oxidize methane to carbon dioxide. Based on data mined from a review of the literature, we determined that the mean methane oxidation rate was 1.90 mg CH₄ m⁻² day⁻¹ and that the main variables controlling this rate were soil water content and inorganic nitrogen in the soils. In contrast, the effects of temperature and pH are minimal. In addition to reviewing the literature, we monitored methane oxidation rates in a temperate forest soil in Korea on a monthly basis for a year, using a static chamber method. The mean oxidation rate was 1.96 mg CH₄ m⁻² day⁻¹ and was positively correlated with nitrate concentration in the soil.     

Responses of oxidation rate and microbial communities to methane in simulated landfill cover soil microcosms

CH4 oxidation capacities and microbial community structures developed in response to the presence of CH4 were investigated in two types of landfill cover soil microcosms, waste soil (fine material in stabilized waste) and clay soil. CH4 emission fluxes were lower in the waste soil cover over the course of the experiment. After exposure to CH4 flow for 120 days, the waste soil developed CH4 oxidation capacity from 0.53 to 11.25-13.48micromol CH4gd.w.(-1)h(-1), which was ten times higher than the clay soil. The topsoils of the two soil covers were observed dried and inhibited CH4 oxidation. The maximum CH4 oxidation rate occurred at the depth of 10-20cm in the waste soil cover (the middle layer), whereas it took place mainly at the depth of 20-30cm in the clay soil cover (the bottom layer). The amounts of the phospholipid fatty acid (PLFA) biomarks 16:1omega8c and 18:1omega8c for type I and II methanotrophs, respectively, showed that type I methanotrophic bacteria predominated in the clay soil, while the type II methanotrophic bacteria were abundant in the waste soil, and the highest population in the middle layer. The results also indicated that a greater active methanotrophic community was developed in the waste soil relative to the clay soil.     

Optimization of diagnostic microarray for application in analysing landfill methanotroph communities under different plant covers

Landfill sites are responsible for 6-12% of global methane emission. Methanotrophs play a very important role in decreasing landfill site methane emissions. We investigated the methane oxidation capacity and methanotroph diversity in lysimeters simulating landfill sites with different plant vegetations. Methane oxidation rates were 35 g methane m-2 day-1 or higher for planted lysimeters and 18 g methane m-2 day-1 or less for bare soil controls. Best methane oxidation, as displayed by gas depth profiles, was found under a vegetation of grass and alfalfa. Methanotroph communities were analysed at high throughput and resolution using a microbial diagnostic microarray targeting the particulate methane monooxygenase (pmoA) gene of methanotrophs and functionally related bacteria. Members of the genera Methylocystis and Methylocaldum were found to be the dominant members in landfill site simulating lysimeters. Soil bacterial communities in biogas free control lysimeters, which were less abundant in methanotrophs, were dominated by Methylocaldum. Type Ia methanotrophs were found only in the top layers of bare soil lysimeters with relatively high oxygen and low methane concentrations. A competetive advantage of type II methanotrophs over type Ia methanotrophs was indicated under all plant covers investigated. Analysis of average and individual results from parallel samples was used to identify general trends and variations in methanotroph community structures in relation to depth, methane supply and plant cover. The applicability of the technology for the detection of environmental perturbations was proven by an erroneous result, where an unexpected community composition detected with the microarray indicated a potential gas leakage in the lysimeter being investigated.     

Culturable psychrotolerant methanotrophic bacteria in landfill cover soil

Methanotrophs closely related to psychrotolerant members of the genera Methylobacter and Methylocella were identified in cultures enriched at 10°C from landfill cover soil samples collected in the period from April to November. Mesophilic methanotrophs of the genera Methylobacter and Methylosinus were found in cultures enriched at 20°C from the same cover soil samples. A thermotolerant methanotroph related to Methylocaldum gracile was identified in the culture enriched at 40°C from a sample collected in May (the temperature of the cover soil was 11.5–12.5°C). In addition to methanotrophs, methylobacteria of the genera Methylotenera and Methylovorus and members of the genera Verrucomicrobium, Pseudomonas, Pseudoxanthomonas, Dokdonella, Candidatus Protochlamydia, and Thiorhodospira were also identified in the enrichment cultures. A methanotroph closely related to the psychrotolerant species Methylobacter tundripaludum (98% sequence identity of 16S rRNA genes with the type strain SV96^T) was isolated in pure culture. The introduction of a mixture of the methanotrophic enrichments, grown at 15°C, into the landfill cover soil resulted in a decrease in methane emission from the landfill surface in autumn (October, November). The inoculum used was demonstrated to contain methanotrophs closely related to Methylobacter tundripaludum SV96.     

Methanotrophic Activity,Abundance, and Diversity in Forested Swamp Pools: Spatiotemporal Dynamics and Influences on Methane Fluxes

Methane oxidation (methanotrophy) in the water column and sediments of forested swamp pools likely control seasonal and annual emission of CH₄ from these systems, but the methanotrophic microbial communities, their activities, locations, and overall impact, is poorly understood. Several techniques including ¹⁴CH₄ oxidation assays, culture-based most probable number (MPN) estimates of methane-oxidizing bacteria (MOB) and protozoan abundance, MOB strain isolation and characterization, and PCR techniques were used to investigate methanotrophy at a forested swamp near Ithaca, New York. The greatest methanotrophic activity and largest numbers of MOB occurred predominantly at the low oxygen sediment/water interface in the water column. Seasonally, methanotrophic activity was very dynamic, ranging from 0.1 to 61.9 μ moles CH₄ d^?1 g^?1 dry sediment, and correlated most strongly with dissolved inorganic carbon (r = 0.896). Incorporation of methanotrophic variables (abundance and activity) into existing CH₄ flux regression models improved model fit, particularly during mid summer when CH₄ fluxes were most dynamic. Annually integrated methane flux and methanotrophic activity measurements indicate that differences in methanotrophic activity at the sediment/water interface likely accounted for differences in the annual CH₄ emission from the field site. Direct isolations of MOB resulted in the repeated isolation of organisms most closely related to Methylomonas methanica S1. A single acidophilic, type II MOB related to Methylocella palustris K was also isolated. Using a PCR-based MPN method and 16S rRNA genome copy number from isolates and control strains, type I and type II MOB were enumerated and revealed type I dominance of the sediment-associated MOB community.     

Methane formation and oxidation in the meromictic oligotrophic Lake Gek-Gel (Azerbaijan)

The production and oxidation of methane and diversity of culturable aerobic methanotrophic bacteria in the water column and upper sediments of the meromictic oligotrophic Lake Gek-Gel (Azerbaijan) were studied by radioisotope, molecular, and microbiological techniques. The rate of methane oxidation was low in the aerobic mixolimnion, increased in the chemocline, and peaked at the depth where oxygen was detected in the water column. Aerobic methanotrophic bacteria of type II belonging to the genus Methylocystis were identified in enrichment cultures obtained from the chemocline. Methane oxidation in the anaerobic water of the monimolimnion was much more intense than in the aerobic zone. However, below 29–30 m methane concentration increased and reached 68 μM at the bottom. The highest rate of methane oxidation under anaerobic conditions was revealed in the upper layer of bottom sediments. The rate of methane oxidation significantly exceeding that of methane production suggests a deep source of methane in this lake.     

Effects of N-fertilisation on CH4 oxidation and production, and consequences for CH4 emissions from microcosms and rice fields

The world's growing human population causes an increasing demand for food, of which rice is one of the most important sources. In rice production nitrogen is often a limiting factor. As a consequence increasing amounts of fertiliser will have to be applied to maximise yields. There is an ongoing discussion on the possible effects of fertilisation on CH₄ emissions. We therefore investigated the effects of N‐fertiliser (urea) on CH₄ emission, production and oxidation in rice microcosms and field experiments. In the microcosms, a substantial but short‐lived reduction of CH₄ emission was observed after N‐addition to 43‐d‐old rice plants. Methane oxidation increased by 45%, demonstrated with inhibitor measurements and model calculations based on stable carbon isotope data (δ¹³CH₄). A second fertilisation applied to 92‐d‐old plants had no effect on CH₄ emission rates. The positive effect of additional N on methanotrophic bacteria was also found in vitro for potential CH₄ oxidation rates in soil and root samples from the microcosm and field experiments, indicated by elevated initial oxidation rates and reduced lag‐phases. Fertilisation did not affect methane production in the microcosms. In the field, the effects were diverse: methane production was inhibited in the topsoil, but stimulated instead in the bulk soil. Stimulation occurred probably in the anaerobic food chain at the level of hydrolytic or fermenting bacteria, because acetate, a methanogenic precursor, increased simultaneously. Combining field, microcosm and laboratory experiments we conclude that any agricultural treatment improving the N‐supply to the rice plants will also be favourable for the CH₄ oxidising bacteria. However, N‐fertilisation had only a transient influence and was counter‐balanced in the field by an elevated CH₄ production. A negative effect of the fertilisation was a transient increase of N₂O emissions from the microcosms. However, integrating over the season the global warming potential (GWP) of N₂O emitted after fertilisation was still negligible compared to the GWP of emitted CH₄.