Structural and functional response of methane-consuming microbial communities to different flooding regimes in riparian soils

Climate change will lead to more extreme precipitation and associated increase of flooding events of soils. This can turn these soils from a sink into a source of atmospheric methane. The latter will depend on the balance of microbial methane production and oxidation. In the present study, the structural and functional response of methane oxidizing microbial communities was investigated in a riparian flooding gradient. Four sites differing in flooding frequency were sampled and soil-physico-chemistry as well as methane oxidizing activities, numbers and community composition were assessed. Next to this, the active community members were determined by stable isotope probing of lipids. Methane consumption as well as population size distinctly increased with flooding frequency. All methane consumption parameters (activity, numbers, lipids) correlated with soil moisture, organic matter content, and conductivity. Methane oxidizing bacteria were present and activated quickly even in seldom flooded soils. However, the active species comprised only a few representatives belonging to the genera Methylobacter, Methylosarcina, and Methylocystis, the latter being active only in permanently or regularly flooded soils.This study demonstrates that soils exposed to irregular flooding harbor a very responsive methane oxidizing community that has the potential to mitigate methane produced in these soils. The number of active species is limited and dominated by one methane oxidizing lineage. Knowledge on the characteristics of these microbes is necessary to assess the effects of flooding of soils and subsequent methane cycling therein.     

Methane oxidation and methylotroph population dynamics in groundwater mesocosms

Extraction of natural gas from unconventional hydrocarbon reservoirs by hydraulic fracturing raises concerns about methane migration into groundwater. Microbial methane oxidation can be a significant methane sink. Here, we inoculated replicated, sand-packed, continuous mesocosms with groundwater from a field methane release experiment. The mesocosms experienced thirty-five weeks of dynamic methane, oxygen and nitrate concentrations. We determined concentrations and stable isotope signatures of methane, carbon dioxide and nitrate and monitored microbial community composition of suspended and attached biomass. Methane oxidation was strictly dependent on oxygen availability and led to enrichment of ¹³C in residual methane. Nitrate did not enhance methane oxidation under oxygen limitation. Methylotrophs persisted for weeks in the absence of methane, making them a powerful marker for active as well as past methane leaks. Thirty-nine distinct populations of methylotrophic bacteria were observed. Methylotrophs mainly occurred attached to sediment particles. Abundances of methanotrophs and other methylotrophs were roughly similar across all samples, pointing at transfer of metabolites from the former to the latter. Two populations of Gracilibacteria (Candidate Phyla Radiation) displayed successive blooms, potentially triggered by a period of methane famine. This study will guide interpretation of future field studies and provides increased understanding of methylotroph ecophysiology.     

Ecological observations on Heterotrophic,methane oxidizing and sulfate reducing bacteria in Pond

Numbers of heterotrophic, methane oxidizing and sulfate reducing bacteria were counted in Lake Vechten. A dynamic distribution pattern was found in the stratified lake. A maximum of heterotrophs (numbers of 10⁹ bact./l) occured in the deepest part of the lake in spring and in the metalimnion during summer-stratification. These bacteria use nearly all available oxygen in the hypolimnion. It was found that the concentration of available organic material and the oxygen tension caused the numbers of heterotrophs in the metalimnion to be high.The maximal numbers of methane oxidizers (numbers of 5.10⁵ bact./l) were found at a depth of maximal methane concentration: the de-oxygenated hypolimnion. Preliminary evidence indicated that these organisms were facultative methane oxidizers and must be regarded as micro-aerophyllics. By oxidizing methane they removed the residual oxygen under the metalimnion.The sulfate reducing bacteria could be observed in the hypolimnion only. Decreased SO _inf4 ^sup–2 concentration and increased numbers, of bacterai were found in the bottom water. An association between the methane oxidizers and the sulfate reducers could be deduced. It was assumed that favourable redox requirements for obligate anaerobic sulfate reducers were the results of the activities of the methane oxidizing bacteria.The dynamic distribution equilibrium of the investigated groups of bacteria was disturbed by the autumn turn-over. The heterotrophic and methane oxidizing bacteria decreased in number at that period and were equally distributed, no sulfate reducers could be detected in the free water of Lake Vechten.     

Comparative studies of products produced from four different biomass samples via deoxy-liquefaction

The objective of this study is to investigate the distribution of products, i.e. gas, liquid oil and char from four different biomass samples (legume straw, corn stalk, cotton stalk and wheat straw) and analysis of the oil for the differences in the hydrocarbon composition with respect to the materials by deoxygenate liquefaction (abbr. deoxy-liquefaction). GC/MS was used to analyze the gas and oil components. According to the similarity of the natural petroleum and bio-petroleum, a new standard for bio-petroleum was established in this paper. The striking characteristic of the bio-petroleum was H/C>1.5, oxygen content <6% and the HHV>40 MJ/kg, containing mainly alkanes, cycloalkanes and aromatic hydrocarbons. In this paper, only the oil produced from legume straw and corn stalk could be called bio-petroleum. The oil derived from different samples contained almost the same compounds, while the relative content varied based on the different content of the main biomass components (lignin and holocellulose). The gaseous products were carbon dioxide, carbon monoxide, methane and hydrogen. In addition, small amount of ethylene, ethane and propane was also observed in gas. The major gas product was carbon dioxide (81.29-86.33%) for all samples.     

Methanogenesis in granular sludge exposed to oxygen

Abstract Substrate competition between methanogenic and facultative bacteria under highly aerobic conditions was investigated in batch experiments. Natural mixed cultures of anaerobic bacteria immobilized in granular sludge were able to concurrently utilize oxygen and produce methane when supplied with ethanol as substrate. The most oxygen tolerant sludge converted 3 to 25% of substrate chemical oxygen demand to methane after 3 days while 23 to 2 mg 1⁻¹ of dissolved oxygen were present in the media. The tolerance of methanogens to oxygen and their coexistence with facultative bacteria were evident even after long periods of oxygen exposure. Eventually, methane oxidizing bacteria developed in the co-culture. The consumption of oxygen by facultative bacteria, creating anaerobic microniches inside the granules, is hypothesized to protect the methanogens.     

Assessment of in-situ bioremediation at a refinery waste-contaminated site and an aviation gasoline contaminated site

A combination of geochemical, microbiological and isotopic methods were used to evaluate in-situ bioremediation of petroleum hydrocarbons at one site contaminated with refinery waste and a second site contaminated with aviation gasoline at Alameda Point, California. At each site, geochemical and microbiological characteristics from four locations in the contaminated zone were compared to those from two uncontaminated background locations. At both sites, the geochemical indicators of in-situbiodegradation includeddepleted soil gas and groundwater oxygen, elevated groundwater alkalinity, and elevated soil gas carbon dioxide and methane in the contaminated zone relative to the background. Radiocarbon content of methane and carbon dioxide measured in soil gas at both sites indicated that they were derived from hydrocarbon contaminant degradation. Direct microscopy of soil core samples using cell wall stains and activity stains, revealed elevated microbial numbers and enhanced microbial activities in contaminated areas relative to background areas, corroborating geochemical findings. While microbial plate counts and microcosm studies using soil core samples provided laboratory evidence for the presence of some microbial activity and contaminant degradation abilities, they did not correlate well with either contaminant location, geochemical, isotopic, or direct microscopy data.     

Harnessing a methane‐fueled,sediment‐free mixed microbial community for utilization of distributed sources of natural gas

Harnessing the metabolic potential of uncultured microbial communities is a compelling opportunity for the biotechnology industry, an approach that would vastly expand the portfolio of usable feedstocks. Methane is particularly promising because it is abundant and energy‐rich, yet the most efficient methane‐activating metabolic pathways involve mixed communities of anaerobic methanotrophic archaea and sulfate reducing bacteria. These communities oxidize methane at high catabolic efficiency and produce chemically reduced by‐products at a comparable rate and in near‐stoichiometric proportion to methane consumption. These reduced compounds can be used for feedstock and downstream chemical production, and at the production rates observed in situ they are an appealing, cost‐effective prospect. Notably, the microbial constituents responsible for this bioconversion are most prominent in select deep‐sea sediments, and while they can be kept active at surface pressures, they have not yet been cultured in the lab. In an industrial capacity, deep‐sea sediments could be periodically recovered and replenished, but the associated technical challenges and substantial costs make this an untenable approach for full‐scale operations. In this study, we present a novel method for incorporating methanotrophic communities into bioindustrial processes through abstraction onto low mass, easily transportable carbon cloth artificial substrates. Using Gulf of Mexico methane seep sediment as inoculum, optimal physicochemical parameters were established for methane‐oxidizing, sulfide‐generating mesocosm incubations. Metabolic activity required >～40% seawater salinity, peaking at 100% salinity and 35 °C. Microbial communities were successfully transferred to a carbon cloth substrate, and rates of methane‐dependent sulfide production increased more than threefold per unit volume. Phylogenetic analyses indicated that carbon cloth‐based communities were substantially streamlined and were dominated by Desulfotomaculum geothermicum. Fluorescence in situ hybridization microscopy with carbon cloth fibers revealed a novel spatial arrangement of anaerobic methanotrophs and sulfate reducing bacteria suggestive of an electronic coupling enabled by the artificial substrate. This system: 1) enables a more targeted manipulation of methane‐activating microbial communities using a low‐mass and sediment‐free substrate; 2) holds promise for the simultaneous consumption of a strong greenhouse gas and the generation of usable downstream products; and 3) furthers the broader adoption of uncultured, mixed microbial communities for biotechnological use.     

全程硝化菌微生物学特性及在水处理领域的应用潜力

全程硝化菌是近期微生物氮循环领域的重大发现之一,引发了对其全球分布、系统发育特征和生理生化特性的广泛关注。本文综述了全程硝化菌在土壤、地表水、废水处理系统等生境的分布规律及影响因子;并从底物亲和力、代谢多样性等方面阐述了其与传统硝化微生物间的竞争互作和生态位分离机制;基于上述特征提出全程硝化菌在水处理领域中的应用前景,可能与其他脱氮微生物如反硝化菌、厌氧氨氧化菌和厌氧甲烷氧化菌等耦合实现在低氨氮、低溶解氧条件下的污水深度脱氮,从而节省能耗并降低温室气体排放。未来研究应继续深入研究全程硝化菌的生理生化特性,评价其生态功能和对氮素地球化学循环的贡献,并探索其在生物水处理等领域的应用潜力。     

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.     

Mimicking the oxygen minimum zones: stimulating interaction of aerobic archaeal and anaerobic bacterial ammonia oxidizers in a laboratory‐scale model system

In marine oxygen minimum zones (OMZs), ammonia‐oxidizing archaea (AOA) rather than marine ammonia‐oxidizing bacteria (AOB) may provide nitrite to anaerobic ammonium‐oxidizing (anammox) bacteria. Here we demonstrate the cooperation between marine anammox bacteria and nitrifiers in a laboratory‐scale model system under oxygen limitation. A bioreactor containing ‘Candidatus Scalindua profunda’ marine anammox bacteria was supplemented with AOA (Nitrosopumilus maritimus strain SCM1) cells and limited amounts of oxygen. In this way a stable mixed culture of AOA, and anammox bacteria was established within 200 days while also a substantial amount of endogenous AOB were enriched. ‘Ca. Scalindua profunda’ and putative AOB and AOA morphologies were visualized by transmission electron microscopy and a C₁₈ anammox [3]‐ladderane fatty acid was highly abundant in the oxygen‐limited culture. The rapid oxygen consumption by AOA and AOB ensured that anammox activity was not affected. High expression of AOA, AOB and anammox genes encoding for ammonium transport proteins was observed, likely caused by the increased competition for ammonium. The competition between AOA and AOB was found to be strongly related to the residual ammonium concentration based on amoA gene copy numbers. The abundance of archaeal amoA copy numbers increased markedly when the ammonium concentration was below 30 μM finally resulting in almost equal abundance of AOA and AOB amoA copy numbers. Massive parallel sequencing of mRNA and activity analyses further corroborated equal abundance of AOA and AOB. PTIO addition, inhibiting AOA activity, was employed to determine the relative contribution of AOB versus AOA to ammonium oxidation. The present study provides the first direct evidence for cooperation of archaeal ammonia oxidation with anammox bacteria by provision of nitrite and consumption of oxygen.     

In situ stimulation of bacterial sulfate reduction in sulfate-limited freshwater lake sediments

Abstract By adding sulfate in the form of solid gypsum, it was possible to transform in situ a predominantly methanogenic sediment ecosystem into a sulfate-reducing one. The concentrations of sulfate, sulfide, methane, acetate, propionate, soluble iron, and manganese were determined in the porewater before and after the transition. Although sulfate was no longer limiting, acetate and propionate continued to accumulate and reached much higher concentrations than under sulfate-limited conditions. Metabolic activities of fermenting bacteria and of sulfate reducers, which belong to the group that incompletely oxidizes organic material, might be responsible for the increased production of volatile fatty acids. The elevated concentrations of soluble Fe(II)²⁺ and Mn(II)²⁺ observed in the porewater stem from iron and manganese compounds which may be reduced chemically by hydrogen sulfide and other microbially produced reducing agents or directly through increased activities of the iron and manganese reducing bacteria. In the horizon with high sulfate-reducing activities the methane concentrations in the porewater were lower than in non-stimulated sediment regions. The shape of the concentration depth profile indicates methane consumption through sulfate reducing processes. The in situ experiment demonstrates the response of a natural microbial ecosystem to fluctuations in the environmental conditions.     

Thermophilic anaerobic microbial communities that transform cellulose into methane (biogas)

The project is devoted to the screening of active anaerobic microbial communities which produce biogas via the decomposition of cellulose in thermophilic conditions (+55°C). Twenty-four samples were isolated from different natural and anthropogenic sources that contain desired microbial organisms. Growth medium was chosen to optimize the conditions for proliferation and selection of cellulolytic and methanogenic microorganisms. During the study of biogas formation dynamics, the most productive communities that remain active during five passages were selected. The biogas composition (methane, carbon dioxide, hydrogen) was investigated by gas chromatography. On average, the methane content in the gas mixture reached 60%. Microscopic studies revealed the presence of various morphotypes of microbial cells; their ratio varied during the stabilization of communities. The significance of the research on the transformation of cellulose into biogas is discussed.     

Functional Groups and Activities of Bacteria in a Highly Acidic Volcanic Mountain Stream and Lake in Patagonia,Argentina

Acidic volcanic waters are naturally occurring extreme habitats that are subject of worldwide geochemical research but have been little investigated with respect to their biology. To fill this gap, the microbial ecology of a volcanic acidic river (pH approximately equal to 0-1.6), Rio Agrio, and the recipient lake Caviahue in Patagonia, Argentina, was studied. Water and sediment samples were investigated for Fe(II), Fe(III), methane, bacterial abundances, biomass, and activities (oxygen consumption, iron oxidation and reduction). The extremely acidic river showed a strong gradient of microbial life with increasing values downstream and few signs of life near the source. Only sulfide-oxidizing and fermentative bacteria could be cultured from the upper part of Rio Agrio. However, in the lower part of the system, microbial biomass and oxygen penetration and consumption in the sediment were comparable to non-extreme aquatic habitats. To characterize similarities and differences of chemically similar natural and man-made acidic waters, our findings were compared to those from acidic mining lakes in Germany. In the lower part of the river and the lake, numbers of iron and sulfur bacteria and total biomass in sediments were comparable to those known from acidic mining lakes. Bacterial abundance in water samples was also very similar for both types of acidic water (around 10(5) mL(-1)). In contrast, Fe(II) oxidation and Fe(III) reduction potentials appeared to be lower despite higher biogenic oxygen consumption and higher photosynthetic activity at the sediment-water interface. Surprisingly, methanogenesis was detected in the presence of high sulfate concentrations in the profundal sediment of Lake Caviahue. In addition to supplementing microbiological knowledge on acidic volcanic waters, our study provides a new view of these extreme sites in the general context of aquatic habitats.     

Ethane oxidation by methane-oxidizing bacteria

The relationship between the rates of methane and ethane oxidation by washed suspensions of methane-oxidizing bacteria has been investigated. Considerable differences between bacterial strains were observed. Two closely related Methylomonas strains which differed in their oxidizing capacity were further investigated. The low ethane oxidation rate of one strain could be strongly stimulated by the addition of oxidizable co-substrates, and the presence of ethane stimulated formate oxidation. The other strain had a much higher ethane oxidation rate and stimulation by co-substrates was negligible.Differences between the levels of dissimilative enzymes in cell-free extracts could not be detected. Attempts to produce extracts with methane mono-oxygenase activity failed. When cells were made permeable with chitosan the results suggested that strains with a low ethane oxidizing capacity obtain the required reductant for the mono-oxygenase from endogenous respiration. In strains with a high ethane oxidation rate, the reductant appears to be derived from oxidation of ethanol or acetaldehyde.     

气态烃诱导下油气微生物数量及功能基因变化特征

【目的】烃类渗漏是地球化学循环中的自然垂直运移现象。油气微生物勘探技术就是通过检测油气微生物在烃类渗漏条件下形成的异常特征进而预测下伏油气藏。然而,上浮烃类比重小,使得油气微生物丰度小,缺乏对油气藏地表烃类与油气微生物的深入认识。【方法】本研究在人工模拟环境下,采用培养法和定量PCR对油气微生物数量和油气功能基因的变化特征进行研究。【结果】人工模拟的不同渗漏环境,在一定的驯化培养周期下分别考察甲烷氧化菌和丁烷氧化菌数量变化特征,甲烷氧化菌与丁烷氧化菌在不同烃源中呈现不同的发育情况;同时,在气态烃高浓度阳性对照和微渗漏条件下,油气指示基因pmo A与bom X基因丰度呈现增长现象,然而经过吹脱实验基因丰度仍能指示出曾经发育的油气微生物,在油气微生物勘探检测分析时间尺度上精度高于数量水平的检测。【结论】本研究考察了土壤中油气微生物的数量和功能基因变化特征,为烃类渗漏与油气微生物之间的相关性研究奠定了基础,为油气微生物勘探提供直接实验依据。     

Development of respirometry methods to assess the microbial activity of thermophilic bioleaching archaea

Respirometry methods have been used for many years to assess the microbial activity of mainly heterotrophic bacteria. Using this technique, the consumption of oxygen and evolution of carbon dioxide for heterotrophic carbon catabolism can be used to assess microbial activity. In the case of autotrophic bioleaching bacteria, carbon dioxide is used as a carbon source resulting in the consumption of both oxygen and carbon dioxide. The use of such respirometry techniques at high temperatures (up to 80 degrees C) for the investigation of bioleaching Archaea, however, poses particular difficulties. At these elevated temperatures, the solubility of oxygen into the liquid phase is particularly poor. This work details specific methods by which high temperature constraints are overcome while monitoring the activity of thermophilic Archaea using a Micro-Oxymax respirometer (Columbus Instruments). The use of elevated headspace oxygen concentrations, in order to overcome low oxygen solubility, is demonstrated as well as the effect of such elevated oxygen concentrations on microbial oxygen consumption rates. The relative rates of oxygen and carbon dioxide consumption are also illustrated during the oxidation of a chalcopyrite concentrate. In addition, this paper details generic methods by which respirometry data can be used to quantify inhibitory effects of a compound such as Na(2)SO(4). The further use of such data in predicting minimum hydraulic reactor retention times for continuous culture bioleaching reactors, as a function of concentration of potentially inhibitory compounds, is also demonstrated.     

Acetylene Reduction Assays for Nitrogen Fixation Freshwaters: a Note of Caution

Lake water samples were observed to transform [14-C]ethylene into water-soluble compounds that were undetectable by conventional acetylene reduction assay procedures. Methane oxidizing bacteria, which are known to be common in freshwaters, appeared to be responsible for this activity. As much as 28 percent of added ethylene has been observed to be transformed and this figure is probably an underestimate. It is suggested that acetylene reduction assays may not be accurately applied to samples containing methane oxidizing bacteria.     

Phylogenetic diversity and activity of anaerobic microorganisms of high-temperature horizons of the Dagang Oilfield (China)

The number of microorganisms of major metabolic groups and the rates of sulfate-reducing and methanogenic processes in the formation waters of the high-temperature horizons of Dagang oilfield have been determined. Using cultural methods, it was shown that the microbial community contained aerobic bacteria oxidizing crude oil, anaerobic fermentative bacteria, sulfate-reducing bacteria, and methanogenic bacteria. Using cultural methods, the possibility of methane production from a mixture of hydrogen and carbon dioxide (H2 + CO2) and from acetate was established, and this result was confirmed by radioassays involving NaH14CO3 and 14CH3COONa. Analysis of 16S rDNA of enrichment cultures of methanogens demonstrated that these microorganisms belong to Methanothermobacter sp. (M. thermoautotrophicus), which consumes hydrogen and carbon dioxide as basic substrates. The genes of acetate-utilizing bacteria were not identified. Phylotypes of the representatives of Thermococcus spp. were found among 16S rDNAs of archaea. 16S rRNA genes of bacterial clones belong to the orders Thermoanaerobacteriales (Thermoanaerobacter, Thermovenabulum, Thermacetogenium, and Coprothermobacter spp.), Thermotogales, Nitrospirales (Thermodesulfovibrio sp.) and Planctomycetales. 16S rDNA of a bacterium capable of oxidizing acetate in the course of syntrophic growth with H2-utilizing methanogens was found at high-temperature petroleum reservoirs for the first time. These results provide further insight into the composition of microbial communities of high-temperature petroleum reservoirs, indicating that syntrophic processes play an important part in acetate degradation accompanied by methane production.     

Improved enrichment culture technique for methane-oxidizing bacteria from marine ecosystems: the effect of adhesion material and gas composition

Cultivation of microbial representatives of specific functional guilds from environmental samples depends largely on the suitability of the applied growth conditions. Especially the cultivation of marine methanotrophs has received little attention, resulting in only a limited number of ex situ cultures available. In this study we investigated the effect of adhesion material and headspace composition on the methane oxidation activity in methanotrophic enrichments obtained from marine sediment. Addition of sterilized natural sediment or alternatively the addition of acid-washed silicon dioxide significantly increased methane oxidation. This positive effect was attributed to bacterial adhesion on the particles via extracellular compounds, with a minimum amount of particles required for effect. As a result, the particles were immobilized, thus creating a stratified environment in which a limited diffusive gas gradients could build up and various microniches were formed. Such diffusive gas gradient might necessitate high headspace concentrations of CH₄ and CO₂ for sufficient concentrations to reach the methane-oxidizing bacteria in the enrichment culture technique. Therefore, high concentrations of methane and carbon dioxide, in addition to the addition of adhesion material, were tested and indeed further stimulated methane oxidation. Use of adhesion material in combination with high concentrations of methane and carbon dioxide might thus facilitate the cultivation and subsequent enrichment of environmentally important members of this functional guild. The exact mechanism of the observed positive effects on methane oxidation and the differential effect on methanotrophic diversity still needs to be explored.     

Wildfire reduces carbon dioxide efflux and increases methane uptake in ponderosa pine forest soils of the southwestern USA

Severe wildfire may cause long-term changes in the soil-atmosphere exchange of carbon dioxide and methane, two gases known to force atmospheric warming. We examined the effect of a severe wildfire 10?years after burning to determine decadal-scale changes in soil gas fluxes following fire, and explored mechanisms responsible for these dynamics. We compared soil carbon dioxide efflux, methane uptake, soil temperature, soil water content, soil O horizon mass, fine root mass, and microbial biomass between a burned site and an unburned site that had similar stand conditions to the burned site before the fire. Compared to the unburned site, soil carbon dioxide efflux was 40% lower and methane uptake was 49% higher at the burned site over the 427-day measurement period. Soil O horizon mass, microbial biomass, fine root mass, and surface soil water content were lower at the burned site than the unburned site, but soil temperature was higher. A regression model showed soil carbon dioxide efflux was more sensitive to changes in soil temperature at the burned site than the unburned site. The relative importance of methane uptake to carbon dioxide efflux was higher at the burned site than the unburned site, but methane uptake compensated for only 1.5% of the warming potential of soil carbon dioxide efflux at the burned site. Our results suggest there was less carbon available at the burned site for respiration by plants and microbes, and the loss of the soil O horizon increased methane uptake in soil at the burned site.