Mol Of Methane Research Articles

Incidence of Predominant Methanogenic Flora in Irritable Bowel Syndrome Patients and Apparently Healthy Controls from North India

Flatulence is a common symptom in patients with irritable bowel syndrome (IBS). This may be due to production of hydrogen by intestinal flora. With the presence of methanogenic flora, 4 mol of hydrogen (H(2)) are used with 1 mol of carbon dioxide (CO(2)) to produce 1 mol of methane (CH(4)), a process greatly reducing the volume of gas in the colon. However, the prevalence of methanogenic flora has not yet been reported in healthy and IBS patients from North India. Therefore, this study was planned. This study was conducted prospectively and included 345 patients with irritable bowel syndrome (fulfilling Rome II criteria) and 254 age- and gender-matched apparently healthy controls. Each subject underwent a hydrogen breath test using 10 g lactulose after an overnight fast. An SC Microlyser from Quintron, USA, was used to measure methane and hydrogen at baseline and at every 30 min for 4 h. Subjects with fasting methane concentration <10 p.p.m. were labeled as low methane producers (LMPs) and > 10 p.p.m. as predominant methane producers (PMPs). The IBS and control groups included 66.78% and 67.53% men, respectively. Mean age in the two groups was 48.52 +/- 30.54 years (range 15-68 years) and 45.67 +/- 30.54 years (range 15-78 years), respectively. Prevalence of predominant methanogenic flora in IBS and control groups was 14.5% (50/345) and 34.6% (88/254), respectively (P < 0.001). Fifty-two out of 254 (20.6 %) were PMPs and 36 out of 254 (14.0%) were LMPs in controls. In contrast to this, IBS patients had 17 out of 354 (4.9%) that were PMPs and 33 out of 345 (9.6%) that were LMPs. Methanogenic flora was significantly lower in IBS patients from North India than in apparently healthy subjects. This may be one of the causes of flatulence in IBS patients.

Dominant sugar utilizers in sediment of Lake Constance depend on syntrophic cooperation with methanogenic partner organisms

Six strains of novel bacteria were isolated from profundal sediment of Lake Constance, a deep freshwater lake in Germany, by direct dilution of the sediment in mineral agar medium containing a background lawn of the hydrogen-scavenging Methanospirillum hungatei as a syntrophic partner. The numbers of colony-forming units obtained after incubation for more than 2 months were in the same range as those of total bacterial counts determined by DAPI staining (up to 10(8) cells per millilitre) suggesting that these organisms were dominant members of the community. Identical dilution series in the absence of methanogenic partners yielded numbers that were lower by two to three orders of magnitude. The dominant bacteria were isolated in defined co-culture with M. hungatei, and were further characterized. Growth was slow, with doubling times of 22-28 h at 28 degrees C. Cells were small, 0.5 x 5 microm in size, Gram-positive, and formed terminal oval spores. At 20 degrees C, glucose was fermented by the co-culture strain BoGlc83 nearly stoichiometrically to 2 mol of acetate and 1 mol of methane plus CO(2). At higher temperatures, also lactate and traces of succinate were formed. Anaerobic growth depended strictly on the presence of a hydrogen-scavenging partner organism and was inhibited by bromoethane sulfonate, which together indicate the need for a syntrophic partnership for this process. Strain BoGlc83 grew also aerobically in the absence of a partner organism. All enzymes involved in ATP formation via glycolysis and acetyl CoA were found, most of them at activities equivalent to the physiological substrate turnover rate. This new type of sugar-fermenting bacterium appears be the predominant sugar utilizer in this environment. The results show that syntrophic relationships can play an important role also for the utilization of substrates which otherwise can be degraded in pure culture.

Sol-gel synthesis of iron catalysers supported on silica and titanium for selectively oxidising methane to formaldehyde

Iron materials supported on silica were prepared by the sol-gel method for evaluating catalytic activity in selective oxidation of methane to formaldehyde. Four catalysts were prepared, one corresponding to the silica support (catalyst 1S), another to the titanium support (catalyst 1T) and two more having 0.5% weight iron loads, one for the silica support (catalyst 2FS) and the last one the titanium support (catalyst 2FT). The higher BET areas were 659 and 850 m2/g for catalysts 1S and 2FS, respectively while catalysts 1T and 2FT displayed areas of 65 and 54 m2/g, respectively. Scanning and transmission electronic microscopy displayed an amorphous structure in the silica-supported materials while titanium-supported materials displayed dense materials having defined structure. X-ray diffraction confirmed the silica’s amorphous structure in 1S and 2FS catalysts and displayed the 1T and 2FT catalysts’ anatase structure. The programmed temperature reduction for the 1S and 2FS catalysts did not display reducible species, while displaying hydrogen consumption peaks related to Fe3O4 reduction to α-Fe via FexO route for 1T and 2FT catalysts. The electronic spectroscopy X-ray photo confirmed the Fe(III) specie as having 710.6 e.V binding energy for both 2FS and 2FT catalysts. Catalytic activity was carried out at atmospheric pressure in a quartz reactor, reaction mixture as CH4/O2/N2 =7.5/1/4 at 400-800°C temperature range. The reaction products were analysed by gas chromatography on Hayesep R and T columns using 5Å molecular screening. The best response for selective oxidation of methane to formaldehyde was displayed by the 2FS catalyst with 3.4% mol methane conversion at 650°C, 11.9% mol formaldehyde selectivity and 0.0211 g HCHO/Kg catalyst yield.

Biological Hydrogen Production Using Chloroform-treated Methanogenic Granules

In fermentative hydrogen production, the low-hydrogen-producing bacteria retention rate limits the suspended growth reactor productivity because of the long hydraulic retention time (HRT) required to maintain adequate bacteria population. Traditional bacteria immobilization methods such as calcium alginate entrapment have many application limitations in hydrogen fermentation, including limited duration time, bacteria leakage, cost, and so on. The use of chloroform-treated anaerobic granular sludge as immobilized hydrogen-producing bacteria in an immobilized hydrogen culture may be able to overcome the limitations of traditional immobilization methods. This paper reports the findings on the performance of fed-batch cultures and continuous cultures inoculated with chloroform-treated granules. The chloroform-treated granules were able to be reused over four fed-batch cultures, with pH adjustment. The upflow reactor packed with chloroform-treated granules was studied, and the HRT of the upflow reactor was found to be as low as 4 h without any decrease in hydrogen production yield. Initial pH and glucose concentration of the culture medium significantly influenced the performance of the reactor. The optimum initial pH of the culture medium was neutral, and the optimum glucose concentration of the culture medium was below 20 g chemical oxygen demand/L at HRT 4 h. This study also investigated the possibility of integrating immobilized hydrogen fermentation using chloroform-treated granules with immobilized methane production using untreated granular sludge. The results showed that the integrated batch cultures produced 1.01 mol hydrogen and 2 mol methane per mol glucose. Treating the methanogenic granules with chloroform and then using the treated granules as immobilized hydrogen-producing sludge demonstrated advantages over other immobilization methods because the treated granules provide hydrogen-producing bacteria with a protective niche, a long duration of an active culture, and excellent settling velocity. This integrated two-stage design for immobilized hydrogen fermentation and methane production offers a promising approach for modifying current anaerobic wastewater treatment processes to harvest hydrogen from the existing systems.

Thermodynamic analyses of hydrogen production from sub-quality natural gas: Part II: Steam reforming and autothermal steam reforming

Part I of this paper analyzed sub-quality natural gas (SQNG) pyrolysis and autothermal pyrolysis. Production of hydrogen via direct thermolysis of SQNGs produces only 2 mol of hydrogen and 1 mol of carbon per mole of methane (CH 4). Steam reforming of SQNG (SRSQNG) could become a more effective approach because the processes produce two more moles of hydrogen via water splitting. A Gibbs reactor unit operation in the AspenPlus™ chemical process simulator was employed to accomplish equilibrium calculations for the SQNG + H 2O and SQNG + H 2O + O 2 systems. The results indicate that water and oxygen inlet flow rates do not significantly affect the decomposition of hydrogen sulfide (H 2S) at temperatures lower than 1000 °C. The major co-product of the processes is carbonyl sulfide (COS) while sulfur dimer (S 2) and carbon disulfide (CS 2) are minor by-products within this temperature range. At higher temperatures (>1300 °C), CS 2 and S 2 become major co-products. No sulfur dioxide (SO 2) or sulfur trioxide (SO 3) is formed during either SRSQNG or autothermal SRSQNG processes, indicating that no environmentally harmful acidic gases are generated.

Hydrogen production for fuel cells by autothermal reforming of methane over sulfide nickel catalyst on a gamma alumina support

Experimental and modelling studies have been conducted on catalytic autothermal reforming (ATR) of methane for hydrogen production over a sulfide nickel catalyst on a gamma alumina support. The experiments are performed with different feedstock under thermally neutral conditions. The results show that the performance of the reformer is dependent on the molar air-to-fuel ratio (A/F), the molar water-to-fuel ratio (W/F) and the flowrate of the feedstock mixture. The optimum conditions for high methane conversion and high hydrogen yield are A/F = 3–3.5, W/F = 2–2.5 and a fuel flowrate below 120–250 l h −1. Under these conditions, a methane conversion of 95–99% and a hydrogen yield of 39–41% on a dry basis can be achieved and 1 mole of methane can produce 1.8 moles of hydrogen at an equilibrium reactor temperature of not exceeding 850 °C. A two-dimensional reactor model is developed to simulate the conversion behaviour of the reactor for further study of the reforming process. The model includes all aspects of the major chemical kinetics and the heat and mass transfer phenomena in the reactor. The predicted results are successfully validated with experimental data.

Evidence of intense archaeal and bacterial methanotrophic activity in the Black Sea water column.

In the northwestern Black Sea, methane oxidation rates reveal that above shallow and deep gas seeps methane is removed from the water column as efficiently as it is at sites located off seeps. Hence, seeps should not have a significant impact on the estimated annual flux of approximately 4.1 x 10(9) mol methane to the atmosphere [W. S. Reeburgh, B. B. Ward, S. C. Wahlen, K. A. Sandbeck, K. A. Kilatrick, and L. J. Kerkhof, Deep-Sea Res. 38(Suppl. 2):S1189-S1210, 1991]. Both the stable carbon isotopic composition of dissolved methane and the microbial community structure analyzed by fluorescent in situ hybridization provide strong evidence that microbially mediated methane oxidation occurs. At the shelf, strong isotope fractionation was observed above high-intensity seeps. This effect was attributed to bacterial type I and II methanotrophs, which on average accounted for 2.5% of the DAPI (4',6'-diamidino-2-phenylindole)-stained cells in the whole oxic water column. At deep sites, in the oxic-anoxic transition zone, strong isotopic fractionation of methane overlapped with an increased abundance of Archaea and Bacteria, indicating that these organisms are involved in the oxidation of methane. In underlying anoxic water, we successfully identified the archaeal methanotrophs ANME-1 and ANME-2, eachof which accounted for 3 to 4% of the total cell counts. ANME-1 and ANME-2 appear as single cells in anoxicwater, compared to the sediment, where they may form cell aggregates with sulfate-reducing bacteria (A. Boetius, K. Ravenschlag, C. J. Schubert, D. Rickert, F. Widdel, A. Giesecke, R. Amann, B. B. Jørgensen, U. Witte, and O. Pfannkuche, Nature 407:623-626, 2000; V. J. Orphan, C. H. House, K.-U. Hinrichs, K. D. McKeegan, and E. F. DeLong, Proc. Natl. Acad. Sci. USA 99:7663-7668, 2002).

Benzoate fermentation by the anaerobic bacterium Syntrophus aciditrophicus in the absence of hydrogen-using microorganisms.

The anaerobic bacterium Syntrophus aciditrophicus metabolized benzoate in pure culture in the absence of hydrogen-utilizing partners or terminal electron acceptors. The pure culture of S. aciditrophicus produced approximately 0.5 mol of cyclohexane carboxylate and 1.5 mol of acetate per mol of benzoate, while a coculture of S. aciditrophicus with the hydrogen-using methanogen Methanospirillum hungatei produced 3 mol of acetate and 0.75 mol of methane per mol of benzoate. The growth yield of the S. aciditrophicus pure culture was 6.9 g (dry weight) per mol of benzoate metabolized, whereas the growth yield of the S. aciditrophicus-M. hungatei coculture was 11.8 g (dry weight) per mol of benzoate. Cyclohexane carboxylate was metabolized by S. aciditrophicus only in a coculture with a hydrogen user and was not metabolized by S. aciditrophicus pure cultures. Cyclohex-1-ene carboxylate was incompletely degraded by S. aciditrophicus pure cultures until a free energy change (DeltaG') of -9.2 kJ/mol was reached (-4.7 kJ/mol for the hydrogen-producing reaction). Cyclohex-1-ene carboxylate, pimelate, and glutarate transiently accumulated at micromolar levels during growth of an S. aciditrophicus pure culture with benzoate. High hydrogen (10.1 kPa) and acetate (60 mM) levels inhibited benzoate metabolism by S. aciditrophicus pure cultures. These results suggest that benzoate fermentation by S. aciditrophicus in the absence of hydrogen users proceeds via a dismutation reaction in which the reducing equivalents produced during oxidation of one benzoate molecule to acetate and carbon dioxide are used to reduce another benzoate molecule to cyclohexane carboxylate, which is not metabolized further. Benzoate fermentation to acetate, CO(2), and cyclohexane carboxylate is thermodynamically favorable and can proceed at free energy values more positive than -20 kJ/mol, the postulated minimum free energy value for substrate metabolism.

Effect of substrate and feed antibiotics on in vitro production of volatile fatty acids and methane in caecal contents of chickens

An attempt was done to identify some factors influencing the caecal fermentation pattern in poultry. Experiments were conducted to study effects of carbohydrate substrates (feed components and supplements) and antibiotics on the formation of volatile fatty acids (VFA) and methane in in vitro incubations of the caecal contents of 7-week-old chickens. Stoichiometry of fermentation differed in cultures with different carbohydrates. Fermentation pattern characterized by high propionate and low acetate production was found in cultures with lactose (0.447 and 0.376 mol/1 mol of VFA produced, respectively) and, to a lesser extent, also in cultures with raffinose. Acetate was the predominant metabolite of starch, pectin and xylan (0.727, 0.773 and 0.685 mol/1 mol of VFA produced, respectively). Fermentation of inulin resulted in high proportion of butyrate, 0.221 mol/1 mol of VFA. Other polysaccharides produced only 0.060–0.111 mol of butyrate per 1 mol of VFA. Oligosaccharides (lactose, raffinose) were fermented more rapidly than polysaccharides. Fermentation of inulin yielded more VFA than fermentation of starch, pectin and xylan. No production of VFA from carboxymethylcellulose was observed. On average, 11 mols of VFA were produced per mol of methane. Lasalocid significantly increased molar proportion of propionate, which is potentially beneficial from the point of view of salmonellae control. The magnitude of improvement, however, was small. Other feed antibiotics tested (avoparcin, bacitracin, lincomycin, spiramycin, tylosin, virginiamycin) produced only non-significant or marginal fermentation shifts. Formation of valerate, isoacids and methane was not significantly influenced by the substrate or by antibiotic treatment.

Syntrophus aciditrophicus sp. nov., a new anaerobic bacterium that degrades fatty acids and benzoate in syntrophic association with hydrogen-using microorganisms.

Strain SBT is a new, strictly anaerobic, gram-negative, nonmotile, non-sporeforming, rod-shaped bacterium that degrades benzoate and certain fatty acids in syntrophic association with hydrogen/formate-using microorganisms. Strain SBT produced approximately 3 mol of acetate and 0.6 mol of methane per mol of benzoate in coculture with Methanospirillum hungatei strain JF1. Saturated fatty acids, some unsaturated fatty acids, and methyl esters of butyrate and hexanoate also supported growth of strain SBT in coculture with Desulfovibrio strain G11. Strain SBT grew in pure culture with crotonate, producing acetate, butyrate, caproate, and hydrogen. The molar growth yield was 17 +/- 1 g cell dry mass per mol of crotonate. Strain SBT did not grow with fumarate, iron(III), polysulfide, or oxyanions of sulfur or nitrogen as electron acceptors with benzoate as the electron donor. The DNA base composition of strain SBT was 43.1 mol% G+C. Analysis of the 16 S rRNA gene sequence placed strain SBT in the delta-subdivision of the Proteobacteria, with sulfate-reducing bacteria. Strain SBT was most closely related to members of the genus Syntrophus. The clear phenotypic and genotypic differences between strain SBT and the two described species in the genus Syntrophus justify the formation of a new species, Syntrophus aciditrophicus.

Surface Organometallic Chemistry of Tin: Reactivity of Two Chiral Organostannics, ((−)-menthyl)Me3Sn and ((−)-menthyl)Me2SnH, toward Silica

Two chiral organostannics, the tetraalkyl ((−)-menthyl)Me3Sn (1) and the hydride ((−)-menthyl)Me2SnH (2), were synthesized. Their reactions with the surface of partially dehydroxylated silica were followed by analysis of evolved gases, IR, and 13C and 119Sn MAS NMR spectroscopic characterizations of the grafted organometallics. With both organostannics, a well-defined chiral surface organometallic compound, ≡Si−O−Sn((−)-menthyl)Me2 (3), is obtained from the reaction of 1 mol of organotin complex with 1 mol of silica surface hydroxyl groups, with formation of 1 mol of methane (from 1) or dihydrogen (from 2). Side reactions leading to nonchiral species, such as ≡Si−O−SnMe3 (4) or involving two surface silanol groups for one organotin molecule, giving for instance (≡Si−O)2Sn((−)-menthyl)Me (5), also occur limiting thus the selectivity of the grafting reaction toward 3. The amount of grafted tin and the nature of the grafted tin species depend on the type of organostannic and on temperature. When using the tetraalkyl ((−)-menthyl)Me3Sn as a precursor, temperatures higher than 100 °C are required in order to graft tin onto the silica surface. Although the amount of grafted tin increases with temperature, the selectivity of the reaction toward 3 remains constant at 85% for temperatures lower than 160 °C and decreases at higher temperatures. With ((−)-menthyl)Me2SnH, tin grafting occurs already at room temperature, but a temperature of 80 °C allows one to graft high tin amounts (4−6 wt % Sn) within a reasonable time, with 95−100% of the grafted tin present as 3 on the silica surface. The reaction is less selective at 100 °C. The chiral surface organometallic species 3 is thermally stable up to 160−200 °C.

Fermentation Patterns in Rabbit Caecal Cultures Supplied with Plant Polysaccharides and Lactate

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Methanogenic transformation of methylfurfural compounds to furfural

The metabolic conversion of 5-methylfurfural and 2-methylfurfural to furfural by a methanogenic bacterium, Methanococcus sp. strain B, was studied. This bacterium was found to use methylfurfural compounds as a growth substrate and to convert them stoichiometrically to furfural. For every mole of methylfurfurals metabolized, almost 1 mol of furfural and 0.7 mol of methane were produced. Several methanogenic bacteria did not carry out this conversion. The metabolic conversion of methylfurfurals is likely to be of value in the anaerobic treatment of methylfurfural-containing wastewaters such as those produced by the paper and pulp industries and oatmeal processing industries. This study adds to the list of the limited number of compounds that are known to serve as electron donors for methanogenesis.

96/04803 Natural gas to syncrude: Making the process pay off

Natural gas is a premium fuel because of its high heating value (210.8 kcal/g{center_dot}mol methane) and clean burning characteristics. Transportation costs from source to market, however, may prevent its use as a fuel. Alternative uses for natural gas include conversions to chemicals and to liquid fuels, which often can be conveniently carried out at the location of the gas source. However, conversions to alternative fuels with lower heating values ({approximately} 156--167 kcal/g{center_dot}carbon atom for hydrocarbons; 170.9 kcal/g{center_dot}mol for methanol) can be rational and economical only under special circumstances. This article presents a brief review and comparison of the salient features of the three major conversion technologies: ammonia, methanol, and syncrude syntheses. Because synthesis gas is a common intermediate in these syntheses, the fundamentals of synthesis gas generation are also included. The objective is to illuminate past progress and to evaluate the possibility for significant advances in these proven commercial technologies.

Measurement of the Carbon Isotopic Composition of Methane Using Helicoidal Laser Eigenstates

The spatially generalized Jones matrix formalism is used to design a laser cavity to make intracavity measurements of the carbon isotopic composition of methane. The method is based on a double optical lever effect for helicoidally polarized eigenstates, permitting to measure successively the ~~CH4 and ~~CH4 concentrations. To choose the probed isotope, one simply tunes the frequency of the laser by Zeeman effect. The experiment exhibits a good agreement with the predictions and permits to measure the ~~CH4/~~CH4 composition ratio of methane with an uncertainty of the order of +0.07% for a sample containing only 6 X 10~~ mole of methane.

Methanogenic conversion of 3-s-methylmercaptopropionate to 3-mercaptopropionate

Anaerobic metabolism of dimethylsulfoniopropionate, an osmolyte of marine algae, in anoxic intertidal sediments involves either cleavage to dimethylsulfide or demethylation to 3-S-methylmercaptopropionate (MMPA) and subsequently to 3-mercaptopropionate. The methanogenic archaea Methanosarcina sp. strain MTP4 (DSM 6636), Methanosarcina acetivorans DSM 2834, and Methanosarcina (Methanolobus) siciliae DSM 3028 were found to use MMPA as a growth substrate and to convert it stoichiometrically to 3-mercaptopropionate. Approximately 0.75 mol of methane was formed per mol of MMPA degraded; methanethiol was not detected as an intermediate. Eight other methanogenic strains did not carry out this conversion. We also studied the conversion of MMPA in anoxic marine sediment slurries. Addition of MMPA (500 (mu)M) resulted in the production of methanethiol which was subsequently converted to methane (417 (mu)M). In the presence of the antibiotics ampicillin, vancomycin, and kanamycin (20 (mu)g/ml each), 275 (mu)M methane was formed from 380 (mu)M MMPA; no methanethiol was formed during these incubations. Only methanethiol was formed from MMPA when 2-bromoethanesulfonate (25 mM) was added to a sediment suspension. These results indicate that in natural environments MMPA could be directly or indirectly a substrate for methanogenic archaea.

Do gas phase adducts form during metalorganic vapour phase epitaxial growth of gallium arsenide?

The new primary arsine hex-5-enylarsine (hexAsH 2) has been synthesized and decomposed in the liquid and gas phases in the presence and absence of trimethylgallium. From the gas phase decomposition by itself in vacuo the main product is hex-1-ene, although smaller amounts of cyclic C 6 products and of conjugated hexadienes are observed. Under growth conditions (1 atm of H 2 > 350°C) the hex-1-ene formed fragments completely because of hexAsH 2 first gives the adduct [Me 3Ga·AshexH 2] but this sequentially loses two moles of methane before giving GaAs, methylenecyclopentane and methane, showing that methylenecyclopentane will necessarily be a product if adduct formation occurs. In the gas phase in vacuo the major C 6 product from Me 3Ga and hexAsH 2 is hex-1-ene although there is a small increase in the proportion of methylenecyclopentane and cyclohexene, suggesting that two pathways, one involving adducts (minor) and the other not, operate. In hydrogen, the major C 6 products are methylenecyclopentane and cyclohexane confirming that adducts can be important, but, since hex-1-ene fragments under these conditions it is not possible to quantify the extent of adduct formation. These results are analysed in terms of the calculated equilibrium constant for adduct formation together with the residence time in the cold zone of the reactor and the probability of bimolecular reactions occurring. It is concluded that comparative studies under a variety of different conditions are essential for a full understanding of the processes occurring during growth.

Mesophilic syntrophic acetate oxidation during methane formation by a triculture at high ammonium concentration

In a mesophilic (37°C) triculture at a high ammonium concentration and pH8, methanogenesis from acetate occurred via syntrophic acetate oxidation. Studies with 14C-labelled substrates showed that the amount of labelled methane formed from 1-14C-labelled acetate was equal to that formed from 2-14C-labelled acetate. Labelled methane was also formed from H14CO3-. These results clearly showed that both the methyl and carboxyl groups of acetate were oxidized to CO2 and that CO2 was reduced to CH4 through hydrogenotrophic methanogenesis. During growth of the triculture, a significant isotopic exchange between the carboxyl group of acetate and bicarbonate occurred. As a result, there was a decrease in the specific activity of 1-14C-acetate, and the production of 14CO2 was slightly higher from 1-14C- than from 2-14C-acetate. For each mole acetate degraded, 0.94 mol methane was formed; 9.2 mmol acetate was metabolized during the 294 days of incubation.

Pyruvate as a substrate for growth and methanogenesis forMethanosarcina barkeri

Methanosarcina barkeri strains (227, MS, and UBS) were tested for their ability to utilize pyruvate for growth and methanogenesis. All three strains grown on methanol required 4–5 weeks of adaptation for growth on pyruvate, whereas they required only 2–3 weeks of adaptation for growth on acetate. The adapted cells had a lag of 3–4 days for growth on acetate and 5–10 days for growth on pyruvate. Equimolar amounts of methane were produced from acetate, whereas 0.6–0.7 mol of methane was produced per mol of pyruvate. The optimal concentration of pyruvate for growth and methanogenesis for all three strains was 100 mM, and doubling times were in the range of 135–170 h.

Methane emission and methane oxidation in land-fill cover soil

The vertical profiles of methane and oxygen concentrations were measured in the cover soil at four sites in a restored and covered landfill. At sites 2 and 3 within the landfill area methane was detectable even to the soil surface and emission of methane occured at these two sites. Measured methane emission rates varied seasonally and appeared to be most influenced by soil water content. On an annual basis methane emissions at these two sites were 495 and 909 mol methane m−2 y−1, respectively. At sites 1 and 4 methane was detected in the cover soil but was not present in the immediate subsurface layer, and emission of methane did not occur. Oxidation of methane by bacteria within the soil profile at these two sites appeared to prevent methane emission from the surface. A methane-oxidising microflora had been enriched in the soils of all four landfill sites, as shown by counts of methanotrophs and methylotrophs garden soil not subjected to elevated methane. Counts of methanotrophs and methylotrophs were generally higher in those soil strata where methane concentrations were greatest. Methane oxidation rates were maximum at soil depths where gradients of methane and oxygen overlapped, usually 10–30 cm depth. The depth integrated rates of methane oxidation were very high at sites 2 and 3, the sites also where methane was emitted from the soil surface. A maximum oxidation rate of 450 mmol CH4 m−2 d−1 was measured at site 3. The data suggested that the microflora in the soil above landfill adapted to the presence of elevated methane concentrations by selection of a more methanotrophic community which was able to rapidly oxidise methane. Optimisation of microbial oxidation of methane by bacteria in landfill cover soil may provide a cheap management strategy to minimise the emissions of methane to the atmosphere from landfill.