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PubMed2001

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 O(2) 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 microM). 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.

Culture Media↗

PubMed2003

Growth and methane oxidation rates of anaerobic methanotrophic archaea in a continuous-flow bioreactor.

Anaerobic methanotrophic archaea have recently been identified in anoxic marine sediments, but have not yet been recovered in pure culture. Physiological studies on freshly collected samples containing archaea and their sulfate-reducing syntrophic partners have been conducted, but sample availability and viability can limit the scope of these experiments. To better study microbial anaerobic methane oxidation, we developed a novel continuous-flow anaerobic methane incubation system (AMIS) that simulates the majority of in situ conditions and supports the metabolism and growth of anaerobic methanotrophic archaea. We incubated sediments collected from within and outside a methane cold seep in Monterey Canyon, Calif., for 24 weeks on the AMIS system. Anaerobic methane oxidation was measured in all sediments after incubation on AMIS, and quantitative molecular techniques verified the increases in methane-oxidizing archaeal populations in both seep and nonseep sediments. Our results demonstrate that the AMIS system stimulated the maintenance and growth of anaerobic methanotrophic archaea, and possibly their syntrophic, sulfate-reducing partners. Our data demonstrate the utility of combining physiological and molecular techniques to quantify the growth and metabolic activity of anaerobic microbial consortia. Further experiments with the AMIS system should provide a better understanding of the biological mechanisms of methane oxidation in anoxic marine environments. The AMIS may also enable the enrichment, purification, and isolation of methanotrophic archaea as pure cultures or defined syntrophic consortia.

5'-Nucleotidase↗

PubMed2005

In vitro study of lipid biosynthesis in an anaerobically methane-oxidizing microbial mat.

The anaerobic oxidation of methane (AOM) is a key process in the global methane cycle, and the majority of methane formed in marine sediments is oxidized in this way. Here we present results of an in vitro 13CH4 labeling study (delta13CH4, approximately 5,400 per thousand) in which microorganisms that perform AOM in a microbial mat from the Black Sea were used. During 316 days of incubation, the 13C uptake into the mat biomass increased steadily, and there were remarkable differences for individual bacterial and archaeal lipid compounds. The greatest shifts were observed for bacterial fatty acids (e.g., hexadec-11-enoic acid [16:1Delta11]; difference between the delta13C at the start and the end of the experiment [Deltadelta13C(start-end)], approximately 160 per thousand). In contrast, bacterial glycerol diethers exhibited only slight changes in delta13C (Deltadelta13C(start-end), approximately 10 per thousand). Differences were also found for individual archaeal lipids. Relatively high uptake of methane-derived carbon was observed for archaeol (Deltadelta13C(start-end), approximately 25 per thousand), a monounsaturated archaeol, and biphytanes, whereas for sn-2-hydroxyarchaeol there was considerably less change in the delta13C (Deltadelta13C(start-end), approximately 2 per thousand). Moreover, an increase in the uptake of 13C for compounds with a higher number of double bonds within a suite of polyunsaturated 2,6,10,15,19-pentamethyleicosenes indicated that in methanotrophic archaea there is a biosynthetic pathway similar to that proposed for methanogenic archaea. The presence of group-specific biomarkers (for ANME-1 and ANME-2 associations) and the observation that there were differences in 13C uptake into specific lipid compounds confirmed that multiple phylogenetically distinct microorganisms participate to various extents in biomass formation linked to AOM. However, the greater 13C uptake into the lipids of the sulfate-reducing bacteria (SRB) than into the lipids of archaea supports the hypothesis that there is autotrophic growth of SRB on small methane-derived carbon compounds supplied by the methane oxidizers.

Anaerobiosis↗

PubMed1971

Production of bacterial cells from methane.

A mixed methane-oxidizing bacterial culture capable of stable and predictable growth in continuous culture was isolated. The culture consisted of two types of gram-negative nonsporulating rods resembling pseudomonads. The culture grew well at 45 C on an inorganic medium without asepsis. Specific metal requirements for Ca(2+), Cu(2+), MoO(4) (2-), Zn(2+), Mn(2+), Mg(2+), and Fe(3+) (or Fe(2+)) were shown. The cells grown in continuous culture contained 11.7 to 12.1% total nitrogen. From an animal nutrition standpoint, the distribution of amino acids was satisfactory. The continuous fermentation was operated over a range of steady-state dilution rates from 0.085 to 0.301 hr(-1). The maximum specific growth rate for the culture, mu(max), was 0.303 hr(-1) (doubling time 2.29 hr). The average yield for all fermentations analyzed was 0.616 g (dry weight of cells per g of methane used and 0.215 g (dry weight) of cells per g of oxygen used. The yields on both methane and oxygen were higher for the oxygen-limited than for the methane-limited fermentations. The maximum productivity attained in the fermentor was 2.39 g (dry weight) of cells per hr per liter at a dilution rate of 0.187 hr(-1) and a cell concentration of 12.8 g (dry weight) of cells per liter. The limit on maximum cell productivity was determined only by the mass transfer rate of oxygen in the fermentor. The simultaneous volumetric mass-transfer coefficients (k(L)a in hr(-1)) for oxygen and methane were determined. The results appear to indicate an oxygen to methane mass-transfer coefficient ratio of approximately 1.4.

Amino Acids↗

PubMed1989

Methane production in ruminants: its effect on the doubly labeled water method.

The doubly labeled water (DLW) technique for measuring CO2 production (rCO2) in free-living animals requires an assessment of the elimination of both 2H and 18O from the body over a long period of time. To calculate rCO2, it is necessary to calculate water turnover (rH2O) from the 2H flux rate. In ruminant animals, the accuracy of this calculation is affected by the loss of 2H in methane. We have quantified the effect of methane production (rCH4) on the 2H flux rate, determined in four sheep given 2H2O. The methane produced was depleted in 2H relative to the urine. A relationship between the enrichment of the methane and urine was established. The ratio of urine to methane enrichment was found on average to be 0.6536, and this value was unaffected by the level of rCH4 but showed some dependence on the absolute concentration of 2H in urine. For this reason, the ratio value obtained from four sheep not given 2H2O was different, a mean of 0.6886 was measured, this ratio was unaffected by changes in the diet supplied to the animals. Computer modeling was used to illustrate the dependence of the isotopically derived value for rCO2 on not only rCH4 but also the magnitude of rCO2 itself. The effect of rCH4 on the DLW method can be predicted from the observed ratio of rCO2 to rCH4 and the value of 0.6536 obtained for the ratio of methane to urine enrichment. With the use of data from several studies at this Institute, a limited range of 10 to 20 was found for rCO2/rCH4 in animals fed at or above maintenance.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

PubMed2003

Atmospheric methane transport near landfill sites.

Methane production rates that have been estimated by a biogas production model (MICROGEN) are combined with an air dispersion model in order to determine the spatial distribution of methane around landfill sites. By utilising dispersion models under extreme atmospheric conditions, a maximum methane concentration around the landfills can be determined. The factors that enhance the maximum methane concentrations, using the meteorological model CALMET in conjunction with the dispersion model CALPUFF, are found to be the wind speed and the percentage of cloud cover. The rates of temperature and pressure variation, as well as the land use category seem to have no effect on the maximum methane concentrations. A rapid reduction of methane concentration is observed a few metres away from the landfill centre while a slower reduction is observed at distances greater than 300 m from it. The performance of this methodology is evaluated by comparing measured concentrations with model predictions.

Air Movements↗

PubMed2006

A model to estimate the methane generation rate constant in sanitary landfills using fuzzy synthetic evaluation.

This paper presents a model using fuzzy synthetic evaluation to estimate the methane generation rate constant, k, for landfills. Four major parameters, precipitation, temperature, waste composition and landfill depth were used as inputs to the model. Whereas, these parameters are known to impact the methane generation, mathematical relationships between them and the methane generation rate constant required to estimate methane generation in landfills, are not known. In addition, the spatial variations of k within a landfill combined with the necessity of site-specific information to estimate its value, makes k one of the most elusive parameters in the accurate prediction of methane generation within a landfill. In this paper, a fuzzy technique was used to develop a model to predict the methane generation rate constant. The model was calibrated and verified using k values from 42 locations. Data from 10 sites were used to calibrate the model and the rest were used to verify it. The model predictions are reasonably accurate. A sensitivity analysis was also conducted to investigate the effect of uncertainty in the input parameters on the generation rate constant.

Bacteria, Anaerobic↗

PubMed2006

Thermophilic anaerobic fermentation of olive pulp for hydrogen and methane production: modelling of the anaerobic digestion process.

The present study investigates the thermophilic biohydrogen and methane production from olive pulp, which is the semi-solid residue coming from the two-phase processing of olives. It focussed on: a) production of methane from the raw olive pulp; b) anaerobic bio-production of hydrogen from the olive pulp; c) subsequent anaerobic treatment of the hydrogen-effluent with the simultaneous production of methane; and d) development of a mathematical model able to describe the anaerobic digestion of the olive pulp and the effluent of hydrogen producing process. Both continuous and batch experiments were performed. The hydrogen potential of the olive pulp amounted to 1.6 mmole H2 per g TS. The methane potential of the raw olive pulp and hydrogen-effluent was as high as 19 mmole CH4 per g TS suggesting that: a) olive pulp is a suitable substrate for methane production; and b) biohydrogen production can be very efficiently coupled with a subsequent step for methane production.

Anaerobiosis↗

PubMed2002

The effect of oilseeds in diets of lactating cows on milk production and methane emissions.

Thirty-six lactating multiparous Holstein cows were assigned to diets that contained 2.3, 4.0, and 5.6% fat for an entire lactation to determine the effect of oilseeds on milk composition, production, and methane emissions. The diets were formulated so that whole cottonseeds and canola oilseeds provided equal amounts of added fat. Methane emissions were measured every 3 mo from two replicates of four cows per treatment using a room tracer approach. Dry matter intakes and yields of milk and FCM were greater for cows fed the diets containing oilseeds. Although the concentration of protein in milk was reduced, yields of both protein and fat tended to be increased by the addition of fat. Within the milk fat, the concentrations of C10, C12, C14:0, and C16:0 were reduced and concentrations of C18, C18:1, and trans-C18:1 were increased in response to dietary oilseeds. In serum, urea-N was increased by the dietary oilseeds. Supplementation of diets with oilseeds did not affect methane emissions but tended to increase the efficiency of milk produced per unit of methane emitted. A 1.7% addition of fat to the control diet from a combination of oilseed types increased yields of milk without reducing methane emission rates. The strategy of using unsaturated fats from oilseeds to substantially reduce methane emissions was ineffective, although yield of milk was increased.

Animal Feed↗

PubMed2004

Methane production by mixed ruminal cultures incubated in dual-flow fermentors.

This study evaluated the effects of dilution rate and forage-to-concentrate ratio on gas production by rumen microbes. Continuous cultures were used to monitor methane production at three liquid dilution rates (3.2, 6.3, or 12.5%/h) and three forage-to-concentrate ratios (70:30, 50:50, or 30:70). Filtered ruminal contents were allowed 6 d of adaptation to diets followed by 7 d of data collection. Forage consisted of pelleted alfalfa and the concentrate mix included ground corn, soybean meal, and a mineral and vitamin premix. The experiment was replicated in a split-plot design. Total volatile fatty acid production averaged 58.0 mmol/d and was not affected by treatment. Molar proportion of acetate increased with increasing forage-to-concentrate ratio. Molar proportion of propionate tended to decrease at dilution rate of 12.5%/h and increased with the medium and low forage-to-concentrate ratio. Culture pH tended to be greater at a dilution rate of 12.5%/h. Methane production that was calculated from stoichiometric equations was not affected by treatments. However, methane production based on methane concentration in fermentor headspace resulted in an interaction effect of treatments. Stoichiometric equations underestimated methane output at higher dilution rates and with high forage diets. Total diet fermentability was lowest at dilution rate of 3.2%/h. Increasing dilution rates increased microbial yield; increasing the proportion of concentrate improved microbial efficiency. Dilution rate and forage-to-concentrate ratio altered the partition of substrate by microbes. Methane production based on actual concentrations differed from values estimated using stoichiometry of end-product appearance.

Animals↗

PubMedDate not supplied

[Intensity of the microbiological processes of the methane cycle in different types of Baltic lakes].

The intensity of the microbiological processes of methane formation (MF) and methane oxidation (MO) was determined in the sediments and water of different types of Baltic lakes. The emission of methane from the lake sediments and methane distribution in the water column of the lakes were studied as functions of the lake productivity and hydrologic conditions. During summers, the intensity of MF in the lake sediments and waters varied from 0.001 to 106 ml CH4/(dm3 day) and from 0 to 3.2 ml CH4/(1 day), respectively, and the intensity of MO in the sediments and water varied from 0 to 11.2 ml CH4/(dm3 day) and from 0 to 1.1 ml CH4/(1 day), respectively. The total methane production (MP) in the lakes varied from 15 to 5000 ml CH4/(m2 day). In anoxic waters, the MP comprised 9-18% of the total PM in the lakes. The consumption of organic carbon for methanogenesis varied from 0.03 to 9.7 g/(m2 day). The role of the methane cycle in the degradation of organic matter in the lakes increased with their productivity.

Baltic States↗

PubMed2002

Novel anaerobic process for the recovery of methane and compost from food waste.

Multi-step sequential batch two-phase anaerobic composting (MUSTAC) process was used to recover methane and composted material from food waste. The MUSTAC process consists of five leaching beds for hydrolysis, acidification and post-treatment, and an upflow anaerobic sludge blanket (UASB) reactor for methane recovery. This process involves the combined methods of sequential batch operation and two-phase anaerobic digestion for simple operation and high efficiency. Rumen microorganisms are inoculated due to their enhanced cellulolytic activity. Each leaching bed is operated in a sequential batch mode. Five leaching beds are operated in a multi-step mode with a two-day interval between degradation stages. Acidified products in the leachate from the leaching beds are converted to methane in the UASB reactor. The MUSTAC process demonstrated that it was capable of removing 84.9% of volatile solids (VS) and converting 85.6% of biochemical methane potential (BMP) into methane at 10.9 kg VS/m3 x d in 10 days. Methane gas production rate was 2.31 m3/m3 x d. The output from the post-treatment of residues in the same leaching bed without troublesome moving met the Korean regulation on compost, indicating that it could be used for soil amendment.

Bacteria, Anaerobic↗

PubMed2003

[Methane oxidation in soil profiles of broad-leaved Korean pine mixed forest in Changbai Mountain].

Samples were collected from dark-brown forest soil in different depths in the virgin broad-leaved/Korean pine mixed forest in Changbai Mountain. Laboratory study on methane oxidation of forest soil revealed the vertical profiles of methane oxidation activity. The maximum methane oxidation occurred at the horizon 5-15 cm below the soil surface. When incubated with 400 ppmv methane, it had the highest uptake rate of 3.3 nmol.h-1.g-1 dw. Soil at horizon below 25 cm had a negligible capacity to oxidize methane. The methane-oxidation capacity of surface soil (0-5 cm) decreased because of the inhibition effect of high concentration NH4+ on methanotrophs.

Methane↗

PubMedDate not supplied

[Methane content in bottom sediments and water mass of the Black Sea].

The methane content in bottom sediments and water column of the Black Sea was determined using various methods of desorption and analysis of gases and various methods of calculating their concentrations. The head-space method with the use of salting out and calculation by internal standard proved to be the most accurate procedure for the analysis of methane concentration in bottom sediments. The methane content in bottom sediments increased with the depth of sediment sampling. In the upper 50-70 cm of shelf sediments, two minimums of methane concentration were revealed; in deep-sea sediments, only one minimum was recorded (in the 20-50 cm horizons). In the water column, methane concentrations slowly grew from the surface to a depth of 150-200 m and abruptly increased to a depth of 700-1200 m, remaining virtually constant in underlying layers. In certain deep-sea regions, peaks of methane content in the 1000-1200 m horizons of the water column were revealed, which were most probably due to local influx of abyssal waters enriched with this gas.

Geologic Sediments↗

PubMedDate not supplied

[Anaerobic methane oxidation and sulfate reduction in bacterial mats of coral-like carbonate structures in the Black Sea].

A detailed study of the processes of anaerobic methane oxidation and sulfate reduction in the bacterial mats occurring on coral-like carbonate structures in the region of methane seeps in the Black Sea, as well as of the phenotypic diversity of sulfate-reducing bacteria developing in this zone, has been performed. The use of the radioisotopic method shows the microbial mat structure to be heterogeneous. The peak activity of the two processes was revealed when a mixture of the upper (dark) and underlying (intensely pink) layers was introduced into an incubation flask, which confirms the suggestion that methanotrophic archaea and sulfate-reducing bacteria closely interact in the process of anaerobic methane oxidation. Direct correlation between the rate of anaerobic methane oxidation and the methane and electron acceptor concentrations in the medium has been experimentally demonstrated. Several enrichment and two pure cultures of sulfate-reducing bacteria have been obtained from the near-bottom water and bacterial mats. Both strains were found to completely oxidize the substrates to CO2 and H2S. The bacteria grow at temperatures ranging from -1 to 18 (24) degrees C, with an optimum in the 10-18 degrees C range, and require the presence of 1.5-2.5% NaCl and 0.07-0.2% MgCl2 x 6H2O. Regarding the aggregate of their phenotypic characteristics (cell morphology, spectrum of growth substrates, the capacity for complete oxidation), the microorganisms isolated have no analogues among the psychrophilic sulfate-reducing bacteria already described. The results obtained demonstrate the wide distribution of psychrophilic sulfate-reducing bacteria in the near-bottom water and bacterial mats covering the coral-like carbonate structures occurring in the region of methane seeps in the Black Sea, as well as the considerable catabolic potential of this physiological group of psychrophilic anaerobes in deep-sea habitats.

Anaerobiosis↗

PubMed2005

Potential for biohydrogen and methane production from olive pulp.

The present study investigates the potential for thermophilic biohydrogen and methane production from olive pulp, which is the semi-solid residue coming from the two-phase processing of olives. It focussed on: a) production of methane from the raw olive pulp, b) anaerobic bio-production of hydrogen from the olive pulp, and c) subsequent anaerobic treatment of the hydrogen-effluent with the simultaneous production of methane. Both continuous and batch experiments were performed. The hydrogen potential of the olive pulp amounted to 1.6 mmole H2 per g TS. The methane potential of the raw olive pulp and hydrogen-effluent was as high as 19 mmole CH4 per g TS. This suggests that olive pulp is an ideal substrate for methane production and it shows that biohydrogen production can be very efficiently coupled with a subsequent step for methane production.

Anaerobiosis↗

PubMed2006

[On-line method for measurement of the carbon isotope ratio of atmospheric methane and its application to atmosphere of Yakela condensed gas field].

An on-line method for measurement of the 13C/12C ratio of methane by a gas chromatography/high-temperature conversion/ isotope ratio mass spectrometry (GC/C/MS) technique was developed. This method is less laborious, more rapid (45 min), of high precision (+/- 0.4 x 10(-3)) and by using a small amount of sample (about 200 mL of atmosphere). Its application to isotopic characterization, and hence methane source identification, was demonstrated by examination of atmosphere sample collected in Yakela condensed gas field, China. The average 13C/12C ratio of atmospheric methane in Yakela field was -45.0 x 10(-3) heavier by 1.2 x 10(-3) -2.0 x 10(-3) than the global average. This is caused by seepage and diffusing of methane from Yakela condensed gas reservoir. The concentrations of atmospheric methane in daytimes are found to be lower than those in nighttimes, and the corresponding 13C/12C ratios in daytimes are lighter compared to those in nighttimes, a phenomena probably caused by the fact that a small part of methane from Yakela condensate reservoir is consumed in soil's surface under sunlight.

Air↗

PubMed1984

Relation of breath methane with obesity and other factors.

In a study of breath methane concentration 43 percent of males and 49 percent of females produced detectable amounts. The proportion of methane producers increased significantly with age in males but not in females. Obesity as measured by the sum of triceps, forearm, suprailiac and subscapular skinfolds was significantly less in producers than non-producers. Amongst methane producers there was also an inverse relationship between skinfold thickness and the breath methane concentration. Breath methane concentrations were lower in males than females. In addition, a higher proportion of methane producers was found amongst secretors of blood group substances (ABH antigen) than non-secretors.

ABO Blood-Group System↗