Production of High-Viscosity Whey-Glucose Broths by a Xanthomonas campestris Strain

Acclimation of a sandy soil to an air-natural gas mixture stimulated the biological oxidation of chloroform to carbon dioxide. Acetylene and methane inhibited chloroform oxidation. Chloroform oxidation continued up to 31 days in the absence of methane. Chloroform oxidation rates increased at chloroform concentrations up to 5 μg g of soil^-1.     

Oxidation and Assimilation of Atmospheric Methane by Soil Methane Oxidizers

The metabolism of atmospheric methane in a forest soil was studied by radiotracer techniques. Maximum (sup14)CH(inf4) oxidation (163.5 pmol of C cm(sup-3) h(sup-1)) and (sup14)C assimilation (50.3 pmol of C cm(sup-3) h(sup-1)) occurred at the A(inf2) horizon located 15 to 18 cm below the soil surface. At this depth, 31 to 43% of the atmospheric methane oxidized was assimilated into microbial biomass; the remaining methane was recovered as (sup14)CO(inf2). Methane-derived carbon was incorporated into all major cell macromolecules by the soil microorganisms (50% as proteins, 19% as nucleic acids and polysaccharides, and 5% as lipids). The percentage of methane assimilated (carbon conversion efficiency) remained constant at temperatures between 5 and 20(deg)C, followed by a decrease at 30(deg)C. The carbon conversion efficiency did not increase at methane concentrations between 1.7 and 1,000 ppm. In contrast, the overall methane oxidation activity increased at elevated methane concentrations, with an apparent K(infm) of 21 ppm (31 nM CH(inf4)) and a V(infmax) of 188 pmol of CH(inf4) cm(sup-3) h(sup-1). Methane oxidizers from soil depths with maximum methanotrophic activity respired approximately 1 to 3% of the assimilated methane-derived carbon per day. This apparent endogenous respiration did not change significantly in the absence of methane. Similarly, the potential for oxidation of atmospheric methane was relatively insensitive to methane starvation. Soil samples from depths above and below the zone with maximum atmospheric methane oxidation activity showed a dramatic increase in the turnover of the methane assimilated (>20 times increase). Physical disturbance such as sieving or mixing of soil samples decreased methane oxidation and assimilation by 50 to 58% but did not alter the carbon conversion efficiency. Ammonia addition (0.1 or 1.0 (mu)mol g [fresh weight](sup-1)) decreased both methane oxidation and carbon conversion efficiency. This resulted in a dramatic decrease in methane assimilation (85 to 99%). In addition, ammonia-treated soil showed up to 10 times greater turnover of the assimilated methane-derived carbon (relative to untreated soil). The results suggest a potential for microbial growth on atmospheric methane. However, growth was regulated strongly by soil parameters other than the methane concentration. The pattern observed for metabolism of atmospheric methane in soils was not consistent with the physiology of known methanotrophic bacteria.     

水稻土中CH4氧化的研究

在实验室条件下研究了水稻土中CH₄氧化的特性.结果表明,在早稻种植前采集的水稻土不能氧化大气中的CH₄,但当所供给的CH₄浓度>10μl·L^-1时,能迅速氧化CH₄,所供给的CH₄浓度越高,氧化CH₄的速度越大.经高浓度(>1000μl·L^-1)的CH₄预培养10d,可使本来不具有氧化大气CH₄能力的土壤氧化大气CH_4.大田CH₄排放通量高的水稻土,氧化CH₄的能力较大.     

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.     

Spatial distribution of methane-oxidizing activity in a flooded rice soil

In a study on spatial distribution of methane oxidation in an unplanted flooded field, methane-oxidizing activity, analysed in soil samples under laboratory conditions, decreased with increasing depth (25 cm and beyond). In a flooded field planted to rice, rates of methane oxidation followed the order : rhizosphere (collected from roots at 10-20 cm depth) > surface soil at (0-1 cm) > subsurface soil at 10-20 cm depth, diagonally 10-15 cm away from the centre of hill. Application of ammonium sulfate and, to a lesser extent, urea to surface, rhizosphere and subsurface soil samples from flooded field planted to rice effected a distinct inhibition of methane oxidation. Nitrification inhibitors (thiourea, sodium thiosulfate and dicyandiamide) were also effective in inhibiting methane oxidation. Both surface and rhizosphere soil samples harbored higher populations of methane-oxidizing bacteria than the subsurface soil. Inhibition of methane oxidation in surface and rhizosphere soil samples concomitant with the suppression of autotrophic ammonium oxidizers by nitrification inhibitors implicates an active involvement of autotrophic ammonium oxidizers in methane oxidation.     

Unexpected stimulation of soil methane uptake as emergent property of agricultural soils following bio‐based residue application

Intensification of agriculture to meet the global food, feed, and bioenergy demand entail increasing re‐investment of carbon compounds (residues) into agro‐systems to prevent decline of soil quality and fertility. However, agricultural intensification decreases soil methane uptake, reducing, and even causing the loss of the methane sink function. In contrast to wetland agricultural soils (rice paddies), the methanotrophic potential in well‐aerated agricultural soils have received little attention, presumably due to the anticipated low or negligible methane uptake capacity in these soils. Consequently, a detailed study verifying or refuting this assumption is still lacking. Exemplifying a typical agricultural practice, we determined the impact of bio‐based residue application on soil methane flux, and determined the methanotrophic potential, including a qualitative (diagnostic microarray) and quantitative (group‐specific qPCR assays) analysis of the methanotrophic community after residue amendments over 2 months. Unexpectedly, after amendments with specific residues, we detected a significant transient stimulation of methane uptake confirmed by both the methane flux measurements and methane oxidation assay. This stimulation was apparently a result of induced cell‐specific activity, rather than growth of the methanotroph population. Although transient, the heightened methane uptake offsets up to 16% of total gaseous CO₂ emitted during the incubation. The methanotrophic community, predominantly comprised of Methylosinus may facilitate methane oxidation in the agricultural soils. While agricultural soils are generally regarded as a net methane source or a relatively weak methane sink, our results show that methane oxidation rate can be stimulated, leading to higher soil methane uptake. Hence, even if agriculture exerts an adverse impact on soil methane uptake, implementing carefully designed management strategies (e.g. repeated application of specific residues) may compensate for the loss of the methane sink function following land‐use change.     

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.     

长白山阔叶红松林不同深度土壤CH4氧化研究

采集长白山阔叶红松林下不同深度的暗棕色森林土壤,在实验室条件下测定其对高低浓度CH4的氧化。结果表明,土壤氧化CH4的能力随深度变化明显;5～15cm土层具有最大CH4氧化活性,在400ppmv CH4浓度下此土层土壤最大氧化速率可达3．3nmolCH4·h^-1·g^-1 dw;25cm以下土层基本没有CH4氧化活性;因0～5cm土层土壤含有高浓度NH4^+抑制了CH4氧化菌的活性,所以此层土壤对CH4吸收能力下降。     

Mechanistic Analysis of Ammonium Inhibition of Atmospheric Methane Consumption in Forest Soils

Methane consumption by forest soil was studied in situ and in vitro with respect to responses to nitrogen additions at atmospheric and elevated methane concentrations. Methane concentrations in intact soil decreased continuously from atmospheric levels at the surface to 0.5 ppm at a depth of 14 cm. The consumption rate of atmospheric methane in soils, however, was highest in the 4- to 8-cm depth interval (2.9 nmol per g of dry soil per day), with much lower activities below and above this zone. In contrast, extractable ammonium and nitrate concentrations were highest in the surface layer (0 to 2 cm; 22 and 1.6 μmol per g of dry soil, respectively), as was potential ammonium-oxidizing activity (19 nmol per g of dry soil per day). The difference in zonation between ammonium oxidation and methane consumption suggested that ammonia-oxidizing bacteria did not contribute significantly to atmospheric methane consumption. Exogenous ammonium inhibited methane consumption in situ and in vitro, but the pattern of inhibition did not conform to expectations based on simple competition between ammonia and methane for methane monooxygenase. The extent of ammonium inhibition increased with increasing methane concentration. Inhibition by a single ammonium addition remained constant over a period of 39 days. In addition, nitrite, the end product of methanotrophic ammonia oxidation, was a more effective inhibitor of methane consumption than ammonium. Factors that stimulated ammonium oxidation in soil, e.g., elevated methane concentrations and the availability of cosubstrates such as formate, methanol, or β-hydroxybutyrate, enhanced ammonium inhibition of methane oxidation, probably as a result of enhanced nitrite production.     

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.     

Chloroform in runoff water—a two-year study in a small catchment in Southeast Sweden

Chloroform concentrations were observed and input and output fluxes estimated over a 2-yr period in a small coniferous catchment (0.22 km²) in southeast Sweden. Water discharge was measured daily, and runoff water was sampled bi-weekly for chloroform analysis. An approximate chloroform budget was calculated, which indicated that the annual output of 6 μg m⁻² yr⁻¹ was approximately six times higher than the input, inferring an internal source of chloroform in the catchment. To the best of our knowledge, neither flux estimates nor mass balances have previously been made for chloroform on a catchment scale, nor have data regarding natural runoff variation with time been gathered. Concentrations of chloroform in runoff were found to be generally high during wet periods, such as spring, but also peaked during summer rain events. The observed pattern suggests that chloroform is formed in surface soil layers and transported to the outlet under high-flow conditions and during dry-period rain events; it is lost through degradation or evaporation during drier periods due to longer soil water residence times. The data suggest that the variation among replicates increases with concentration; this emphasizes the need to know what the degree of on-site variation is, so one can collect a sufficient number of replicates to permit detection of spatial or temporal changes.     

植物在CH4产生、氧化和排放中的作用

综合评述了植物对CH4产生、内源CH4氧化和CH4排放的影响．不同植物释放根系分泌物能力的不同是造成CH4产生量差异的主要原因。而植物不同生育期分泌分泌物能力的差异是造成季节性变化的关键．植物泌O2能力的高低和季节性变化通过影响内源CH4的氧化来改变CH4的排放数量．植物问通气组织数量和密度的差异及其随生育期的变化,通过影响对CH4的传输能力来改变CH4的排放量．因此,植物排放CH4的通量及其季节性变化规律是由植物根系分泌分泌物能力、分泌O2能力和传输CH4能力综合决定的．     

Zero methane emission bogs: extreme rhizosphere oxygenation by cushion plants in Patagonia

? Vascular wetland plants may substantially increase methane emissions by producing root exudates and easily degradable litter, and by providing a low-resistance diffusion pathway via their aerenchyma. However, model studies have indicated that vascular plants can reduce methane emission when soil oxygen demand is exceeded by oxygen released from roots. Here, we tested whether these conditions occur in bogs dominated by cushion plants. ? Root-methane interactions were studied by comparing methane emissions, stock and oxygen availability in depth profiles below lawns of either cushion plants or Sphagnum mosses in Patagonia. ? Cushion plants, Astelia pumila and Donatia fascicularis, formed extensive root systems up to 120 cm in depth. The cold soil (< 10°C) and highly decomposed peat resulted in low microbial activity and oxygen consumption. In cushion plant lawns, high soil oxygen coincided with high root densities, but methane emissions were absent. In Sphagnum lawns, methane emissions were substantial. High methane concentrations were only found in soils without cushion plant roots. ? This first methane study in Patagonian bog vegetation reveals lower emissions than expected. We conclude that cushion plants are capable of reducing methane emission on an ecosystem scale by thorough soil and methane oxidation.     

Long-term effects of mineral versus organic fertilizers on activity and structure of the methanotrophic community in agricultural soils

Agricultural practices, such as mineral nitrogen fertilization, have an impact on the soil's ability to oxidize methane, but little is known about the shifts in the methanotrophic community composition associated with these practices. Therefore, the long-term effect of both mineral (NH4NO3) and organic (manure and GFT-compost) fertilizer applications on the soil methanotrophic community activity and structure were investigated. Both high and low affinity methane oxidation rates were lower in the soil treated with mineral fertilizer compared to the other soils. An enhanced nitrate concentration was observed in the mineral fertilized soil but nitrate did not show a direct affect on the high affinity methane oxidation. In contrast, the low affinity methane oxidation was slowed down by increased nitrate concentrations, which suggests a direct effect of nitrate on low affinity methane oxidation. Denaturing gradient gel electrophoresis (DGGE) analysis of 16S rRNA gene fragments specific for methanotrophs revealed a distinct community between the mineral and organic fertilized soils as extra Type I methanotrophic bands (phylotypes) became visible in the organic fertilized soils. These phylotypes were not visible in the patterns of the added organic fertilizers suggesting an indirect effect of the organic fertilizers on the methanotrophic community. Additionally, a molecular analysis was performed after the low affinity methane oxidation test. The enhanced methane concentrations used in the test enriched certain low affinity methanotrophs in the organic fertilized soils but not in the mineral fertilized soil. Supporting the molecular and functional observations, fatty acids characteristic for methanotrophs were less abundant in the soil treated with mineral fertilizer compared to the soil treated with compost. In conclusion, the function and molecular and chemical composition of the methanotrophic community are all altered in soil fertilized with mineral fertilizer.     

Methanol Improves Methane Uptake in Starved Methanotrophic Microorganisms

Methanotrophs in enrichment cultures grew and sustained atmospheric methane oxidation when supplied with methanol. If they were not supplied with methanol or formate, their atmospheric methane oxidation came to a halt, but it was restored within hours in response to methanol or formate. Indigenous forest soil methanotrophs were also dependent on a supply of methanol upon reduced methane access but only when exposed to a methane-free atmosphere. Their immediate response to each methanol addition, however, was to shut down the oxidation of atmospheric methane and to reactivate atmospheric methane oxidation as the methanol was depleted.     

A methane-driven microbial food web in a wetland rice soil

Methane oxidation is a key process controlling methane emission from anoxic habitats into the atmosphere. Methanotrophs, responsible for aerobic methane oxidation, do not only oxidize but also assimilate methane. Once assimilated, methane carbon may be utilized by other organisms. Here we report on a microbial food web in a rice field soil driven by methane. A thin layer of water-saturated rice field soil was incubated under opposing gradients of oxygen and (13)C-labelled methane. Bacterial and eukaryotic communities incorporating methane carbon were analysed by RNA-stable isotope probing (SIP). Terminal restriction fragment length polymorphism (T-RFLP) and cloning showed that methanotrophs were the most prominent group of bacteria incorporating methane carbon. In addition, a few Myxobacteria-related sequences were obtained from the 'heavy' rRNA fraction. Denaturing gradient gel electrophoresis (DGGE) targeting 18S rRNA detected various groups of protists in the 'heavy' rRNA fraction including naked amoeba (Lobosea and Heterolobosea), ciliates (Colpodea) and flagellates (Cercozoa). Incubation of soil under different methane concentrations in air resulted in the development of distinct protozoan communities. These results suggest that methane carbon is incorporated into non-methanotrophic pro- and microeukaryotes probably via grazing, and that methane oxidation is a shaping force of the microeukaryotic community depending on methane availability.     

温度对甲烷产生和氧化的影响

综述了温度对土壤产甲烷和氧化甲烷的影响及其机制．温度主要通过土壤中产甲烷菌的优势菌发生更替来改变土壤的产甲烷能力．较高温条件下产甲烷菌以乙酸和H2／CO2都能利用的甲烷八叠球菌(Methanosarcinaceae)为主,使得土壤处于较高的产甲烷状态．较低温条件下产甲烷菌以只能利用乙酸的甲烷毛菌(Methanosaetaceae)为主,土壤形成甲烷的能力相对较弱．温度提高可以显著地增加甲烷的产生,Q10为1．5-28,平均4．1,但是温度效应明显受控于底物浓度,提高底物浓度降低了产甲烷菌对底物的亲和力,相应地增加了度效应,因此在较低温条件下提高底物浓度可以促进甲烷的产生．温度对大气甲烷氧化的影响弱于产甲烷,甲烷氧化菌较少受温度变化的影响,即便在较低温条件下,土壤也具有一定的氧化大气甲烷能力,原因尚不清楚,可能与甲烷氧化菌对大气甲烷具有较高的亲和力有关,有待进一步研究．     

Biogeochemical processes of methane cycle in the soils, swamps and lakes of Western Siberia

The biogeochemical processes of methane production and oxidation were studied in the upper horizons of tundra and taiga soils and of raised bogs and lake bottom sediments nearby the Tarkosalinsk gas field in western Siberia. Both in dry and water-logged soils, the total methane concentration (in soil particles and gaseous phase) was an order of magnitude higher than in the soil gaseous phase alone (22 and 1.1 nl/cm3, respectively). In bogs and lake bottom sediments, methane concentration was as high as 11 microliters/cm3. Acetate was the major precursor of the newly formed methane. The rate of aceticlastic methanogenesis reached 55 ng C/(cm3 day), whereas that of autotrophic methanogenesis was an order of magnitude lower. The most active methane production and oxidation were observed in bogs and lake sediments where the delta 13C values of CO2 were inversely related to the intensity of bacterial methane oxidation. Methane diffusing from bogs and lake bottom sediments showed delta 13C values ranging from -78 to -47@1000, whereas the delta 13C value of carbon dioxide ranged from -18 to -6@1000. In these ecosystems, methane emission comprised from 3 to 206 mg CH4/(m2 day). Conversely, the dry and water-logged soils of tundra and taiga took up atmospheric methane at a rate varying from 0.3 to 5.3 mg CH4/(m2 day). Methane consumption in soils was of biological rather than of adsorptive nature. This was confirmed by the radioisotopic method and chamber experiments, in which weighting of methane carbon was observed (the delta 13C value changed from -51 to -41@1000).     

Microbial removal of halogenated methanes, ethanes, and ethylenes in an aerobic soil exposed to methane

Abstract Contamination of ground water with halogenated aliphatic hydrocarbons threatens this source of drinking water. In order to study microbial processes that may enhance the removal of these compounds, Lincoln fine sand was exposed to an atmosphere containing methane (4%) to enrich microorganisms capable of growth on this gaseous hydrocarbon. The methane-enriched soil was then tested to determine whether the enriched microbes could remove seven halocarbons from aqueous solution. Removal of dichloromethane. trans -1,2-dichloroethylene, chloroform, 1,2-dichloroethane, trichloroethylene, and 1,1,1-trichloroethane was significantly different in methane-enriched soil compared to non-enriched soil (ANOVA, 95% significance level). Tetrachloroethylene was not removed. Autoclaving the methane-enriched soil inhibited completely the removal of all the compounds. Once the soil was enriched with methane, its presence in the headspace was not required for removal of several of the compounds but methane was required for their complete removal. These results suggest that methane stimulation of microbial communities may be an alternative treatment technology for bioremediation of contaminated subsurface soils and ground water.