Metabolic responses of novel cellulolytic and saccharolytic agricultural soil Bacteria to oxygen

Cellulose is the most abundant biopolymer in terrestrial ecosystems and is degraded by microbial communities in soils. However, relatively little is known about the diversity and function of soil prokaryotes that might participate in the overall degradation of this biopolymer. The active cellulolytic and saccharolytic Bacteria in an agricultural soil were evaluated by 16S rRNA ¹³C‐based stable isotope probing. Cellulose, cellobiose and glucose were mineralized under oxic conditions in soil slurries to carbon dioxide. Under anoxic conditions, these substrates were converted primarily to acetate, butyrate, carbon dioxide, hydrogen and traces of propionate and iso‐butyrate; the production of these fermentation end‐products was concomitant with the apparent reduction of iron(III). [¹³C]‐cellulose was mainly degraded under oxic conditions by novel family‐level taxa of the Bacteroidetes and Chloroflexi, and a known family‐level taxon of Planctomycetes, whereas degradation under anoxic conditions was facilitated by the Kineosporiaceae (Actinobacteria) and cluster III Clostridiaceae and novel clusters within Bacteroidetes. Active aerobic sub‐communities in oxic [¹³C]‐cellobiose and [¹³C]‐glucose treatments were dominated by Intrasporangiaceae and Micrococcaceae (Actinobacteria) whereas active cluster I Clostridiaceae (Firmicutes) were prevalent in anoxic treatments. A very large number (i.e. 28) of the detected taxa did not closely affiliate with known families, and active Archaea were not detected in any of the treatments. These collective findings suggest that: (i) a large uncultured diversity of soil Bacteria was involved in the utilization of cellulose and products of its hydrolysis, (ii) the active saccharolytic community differed phylogenetically from the active cellulolytic community, (iii) oxygen availability impacted differentially on the activity of taxa and (iv) different redox guilds (e.g. fermenters and iron reducers) compete or interact during cellulose degradation in aerated soils.     

Redox fluctuation structures microbial communities in a wet tropical soil

Frequent high-amplitude redox fluctuation may be a strong selective force on the phylogenetic and physiological composition of soil bacterial communities and may promote metabolic plasticity or redox tolerance mechanisms. To determine effects of fluctuating oxygen regimens, we incubated tropical soils under four treatments: aerobic, anaerobic, 12-h oxic/anoxic fluctuation, and 4-day oxic/anoxic fluctuation. Changes in soil bacterial community structure and diversity were monitored with terminal restriction fragment length polymorphism (T-RFLP) fingerprints. These profiles were correlated with gross N cycling rates, and a Web-based phylogenetic assignment tool was used to infer putative community composition from multiple fragment patterns. T-RFLP ordinations indicated that bacterial communities from 4-day oxic/anoxic incubations were most similar to field communities, whereas those incubated under consistently aerobic or anaerobic regimens developed distinctly different molecular profiles. Terminal fragments found in field soils persisted either in 4-day fluctuation/aerobic conditions or in anaerobic/12-h treatments but rarely in both. Only 3 of 179 total fragments were ubiquitous in all soils. Soil bacterial communities inferred from in silico phylogenetic assignment appeared to be dominated by Actinobacteria (especially Micrococcus and Streptomycetes), "Bacilli," "Clostridia," and Burkholderia and lost significant diversity under consistently or frequently anoxic incubations. Community patterns correlated well with redox-sensitive processes such as nitrification, dissimilatory nitrate reduction to ammonium (DNRA), and denitrification but did not predict patterns of more general functions such as N mineralization and consumption. The results suggest that this soil's indigenous bacteria are highly adapted to fluctuating redox regimens and generally possess physiological tolerance mechanisms which allow them to withstand unfavorable redox periods.     

The effect of sediment redox potential and soil acidity on nitrogen uptake,anaerobic root respiration,and growth of rice (Oryza sativa)

Summary Oryza sativa Loisel cultivar Mars., a common lowland rice variety was grown under controlled soil redox conditions (Eh) and acidity (pH). The effect of two variables (Eh and pH) on growth, anaerobic root respiration, and uptake of added labelled nitrogen, was investigated. Plant growth, estimated by dry weight showed significantly higher growth under reducing sediment redox potentials (−200 mV and 0 mV) and at a soil pH of 6.5 Using the activity of the inducible enzyme alcohol dehydrogenase (ADH) as an indicator of anaerobic root respiration, a decrease in redox potential resulted in an increase in root ADH. However, growth paralled increases in anaerobic root respiration suggesting nitrogen transformation in the soil to be a primary parameter governing growth. Labelled nitrogen uptake which was greater under anaerobic conditions apparently led to greater growth of lowland rice in the highly reduced or anaerobic soil treatments.     

Microbial communities acclimate to recurring changes in soil redox potential status

Rapidly fluctuating environmental conditions can significantly stress organisms, particularly when fluctuations cross thresholds of normal physiological tolerance. Redox potential fluctuations are common in humid tropical soils, and microbial community acclimation or avoidance strategies for survival will in turn shape microbial community diversity and biogeochemistry. To assess the extent to which indigenous bacterial and archaeal communities are adapted to changing in redox potential, soils were incubated under static anoxic, static oxic or fluctuating redox potential conditions, and the standing (DNA‐based) and active (RNA‐based) communities and biogeochemistry were determined. Fluctuating redox potential conditions permitted simultaneous CO₂ respiration, methanogenesis, N₂O production and iron reduction. Exposure to static anaerobic conditions significantly changed community composition, while 4‐day redox potential fluctuations did not. Using RNA : DNA ratios as a measure of activity, 285 taxa were more active under fluctuating than static conditions, compared with three taxa that were more active under static compared with fluctuating conditions. These data suggest an indigenous microbial community adapted to fluctuating redox potential.     

Redox Fluctuation Structures Microbial Communities in a Wet Tropical Soil

Frequent high-amplitude redox fluctuation may be a strong selective force on the phylogenetic and physiological composition of soil bacterial communities and may promote metabolic plasticity or redox tolerance mechanisms. To determine effects of fluctuating oxygen regimens, we incubated tropical soils under four treatments: aerobic, anaerobic, 12-h oxic/anoxic fluctuation, and 4-day oxic/anoxic fluctuation. Changes in soil bacterial community structure and diversity were monitored with terminal restriction fragment length polymorphism (T-RFLP) fingerprints. These profiles were correlated with gross N cycling rates, and a Web-based phylogenetic assignment tool was used to infer putative community composition from multiple fragment patterns. T-RFLP ordinations indicated that bacterial communities from 4-day oxic/anoxic incubations were most similar to field communities, whereas those incubated under consistently aerobic or anaerobic regimens developed distinctly different molecular profiles. Terminal fragments found in field soils persisted either in 4-day fluctuation/aerobic conditions or in anaerobic/12-h treatments but rarely in both. Only 3 of 179 total fragments were ubiquitous in all soils. Soil bacterial communities inferred from in silico phylogenetic assignment appeared to be dominated by Actinobacteria (especially Micrococcus and Streptomycetes), “Bacilli,” “Clostridia,” and Burkholderia and lost significant diversity under consistently or frequently anoxic incubations. Community patterns correlated well with redox-sensitive processes such as nitrification, dissimilatory nitrate reduction to ammonium (DNRA), and denitrification but did not predict patterns of more general functions such as N mineralization and consumption. The results suggest that this soil's indigenous bacteria are highly adapted to fluctuating redox regimens and generally possess physiological tolerance mechanisms which allow them to withstand unfavorable redox periods.     

Ex situ slurry phase bioremediation of chrysene contaminated soil with the function of metabolic function: process evaluation by data enveloping analysis (DEA) and Taguchi design of experimental methodology (DOE)

Bioremediation of chrysene in soil matrix was evaluated in soil slurry phase bioreactor in conjugation with metabolic functions (aerobic, anoxic and anaerobic), microenvironment (single and mixed) conditions and nature of mixed consortia (native/resident mixed microflora and bioaugmented inoculum). Twelve experiments were operated independently in agitated-batch reactor keeping all other operating conditions constant (substrate loading rate--0.084 g chrysene/kg soil-day; soil loading rate--10 kg soil/m(3)-day (3:25 soil water ratio); operating temperature--35+/-2 degrees C). Data envelopment analysis (DEA) procedure was employed to analyze the performance of experimental variations in terms of chrysene degradation and pH. The efficacy of anoxic metabolism over the corresponding aerobic and anaerobic metabolic functions was documented. Aerobic metabolic function showed effective degradation capability under mixed microenvironment after augmentation with anaerobic inoculum. Anaerobic metabolic function showed lowest degradation potential. Application of bioaugmentation showed positive influence on the chrysene degradation rate. Design of experimental methodology (DOE) by Taguchi approach was applied to evaluate the effect of four selected factors (native soil microflora, microenvironment, metabolic function and bioaugmentation) on the chrysene degradation process. The optimized factors derived from analysis depicted the requirement of native soil microflora under anoxic metabolic function using mixed microenvironment after augmenting with anaerobic inoculum for achieving effective chrysene degradation efficacy.     

Redox Fluctuations Frame Microbial Community Impacts on N-cycling Rates in a Humid Tropical Forest Soil

Fluctuating soil redox regimes may facilitate the co-occurrence of microbial nitrogen transformations with significantly different sensitivities to soil oxygen availability. In an upland humid tropical forest, we explored the impact of fluctuating redox regimes on gross nitrogen cycling rates and microbial community composition. Our results suggest that the rapidly fluctuating redox conditions that characterize these upland soils allow anoxic and oxic N processing to co-occur. Gross nitrogen mineralization was insensitive to soil redox fluctuations. In contrast, nitrifiers in this soil were directly affected by low redox periods, yet retained some activity even after 3–6 weeks of anoxia. Dissimilatory nitrate reduction to ammonium (DNRA) was less sensitive to oxygen exposure than expected, indicating that the organisms mediating this reductive process were also tolerant of unfavorable (oxic) conditions. Denitrification was a stronger sink for NO₃⁻ in consistently anoxic soils than in variable redox soils. Microbial biomass and community composition were maintained with redox fluctuation, but biomass decreased and composition changed under static oxic and anoxic soil regimes. Bacterial community structure was significantly correlated with rates of nitrification, denitrification and DNRA, suggesting that redox-control of soil microbial community structure was an important determinant of soil N-cycling rates. Specific nitrogen cycling functional groups in this environment (such as nitrifiers, DNRA organisms, and denitrifiers) appear to have adapted to nutrient resources that are spatially and temporally variable. In soils where oxygen is frequently depleted and re-supplied, characteristics of microbial tolerance and resilience can frame N cycling patterns.     

Comparative Assessment of the Aerobic and Anaerobic Microfloras of Earthworm Guts and Forest Soils

Aerobic and anaerobic microbial potentials of guts from earthworms (Lumbricus rubellus Hoffmeister and Octolasium lacteum (Oerl.)) collected from a beech forest were evaluated. On the basis of enumeration studies, microbes capable of growth under both aerobic and anaerobic conditions were more numerous in the earthworm intestine than in the beech forest soil from which the worms were obtained. The intestine of worms displayed nearly equivalent aerobic and anaerobic microbial growth potentials; in comparison, soils displayed greater aerobic than anaerobic microbial growth potentials. Hence, the ratio of microbes capable of growth under obligately anaerobic conditions to those capable of growth under aerobic conditions was higher with the worm intestine than with the soil. Process level studies corroborated these population differentials: (i) under anaerobic conditions, worm gut homogenates consumed glucose, cellobiose, or ferulate more readily than did soil homogenates; and (ii) under aerobic conditions, worm gut homogenates consumed cellobiose or oxygen more readily than did soil homogenates. Collectively, these results reinforce the general concept that the earthworm gut is not microbiologically equivalent to soil and also suggest that the earthworm gut might constitute a microhabitat enriched in microbes capable of anaerobic growth and activity.     

Influence of Microbial Composition on the Carbon Storage Capacity of the Mangrove Soil at the Land-ocean Boundary in the Sundarban Mangrove Ecosystem,India

Spatiotemporal assessment of the mangrove soil of Indian Sundarban revealed that decomposition rate of the organic matter was significantly lower in the anoxic condition than that of the oxic condition. Higher degree of enzyme activity in the oxic soil than the anoxic condition suggested that slower biomineralization in anoxic condition would facilitate long-term storage of organic matter in that particular ecosystem. Microbial population of nitrifying bacteria, phosphate solubilizing bacteria, cellulose degrading bacteria and fungi showed significant reduction in anoxic incubation than that in oxic incubation. In contrary, sulfate reducing bacteria and free living N2 fixing bacteria showed higher population in anoxic incubation indicating their preference for anaerobic condition. Soil CO₂ emission rate decreased with the increase in anoxicity and was largely dependent on the soil redox potential, organic carbon and microbial population of the mangrove soil.     

The leucine incorporation method estimates bacterial growth equally well in both oxic and anoxic lake waters.

Bacterial biomass production is often estimated from incorporation of radioactively labeled leucine into protein, in both oxic and anoxic waters and sediments. However, the validity of the method in anoxic environments has so far not been tested. We compared the leucine incorporation of bacterial assemblages growing in oxic and anoxic waters from three lakes differing in nutrient and humic contents. The method was modified to avoid O(2) contamination by performing the incubation in syringes. Isotope saturation levels in oxic and anoxic waters were determined, and leucine incorporation rates were compared to microscopically observed bacterial growth. Finally, we evaluated the effects of O(2) contamination during incubation with leucine, as well as the potential effects of a headspace in the incubation vessel. Isotope saturation occurred at a leucine concentration of above about 50 nM in both oxic and anoxic waters from all three lakes. Leucine incorporation rates were linearly correlated to observed growth, and there was no significant difference between oxic and anoxic conditions. O(2) contamination of anoxic water during 1-h incubations with leucine had no detectable impact on the incorporation rate, while a headspace in the incubation vessel caused leucine incorporation to increase in both anoxic and O(2)-contaminated samples. The results indicate that the leucine incorporation method relates equally to bacterial growth rates under oxic and anoxic conditions and that incubation should be performed without a headspace.     

Enhanced nutrient removal in three types of step feeding process from municipal wastewater

An anoxic/oxic step feeding process was improved to enhance nutrient removal by reconfiguring the process into (1) anaerobic/anoxic/oxic step feeding process or (2) modified University of Capetown (UCT) step feeding process. Enhanced nitrogen and phosphorus removal and optimized organics utilization were obtained simultaneously in the modified UCT type with both internal and sludge recycle ratios of 75% as well as anaerobic/anoxic/oxic volume ratio of 1:3:6. Specifically, the UCT configuration and optimized operational conditions lead to the enrichment of denitrifying phosphorus removal microorganisms and achieved improved anaerobic P-release and anoxic P-uptake activities, which were beneficial to the denitrifying phosphorus removal activities and removal efficiencies. Due to high mixed liquor suspended solid and uneven distributed dissolved oxygen, 35% of total nitrogen was eliminated through simultaneous nitrification and denitrification process in aerobic zones. Moreover, 62 ± 6% of influent chemical oxygen demands was involved in the denitrification or phosphorus release processes.     

Microbial Aldicarb Transformation in Aquifer, Lake, and Salt Marsh Sediments

The microbial transformation of [N-methyl-(sup14)C]aldicarb, a carbamate pesticide, occurred in aquifer, lake, and salt marsh sediments. Microbial degradation of aldicarb took place within 21 days in aquifer sediments from sites previously exposed to aldicarb (Jamesport, Long Island, N.Y.) but did not occur in sediments which were not previously exposed (Connetquot State Park, Long Island, N.Y.). At the Jamesport sites, higher aldicarb transformation rates occurred in deep, anoxic sediments than in shallow, oxic sediments. There was a significant negative relationship (P < 0.05) between transformation rates and ambient dissolved O(inf2) levels. Aldicarb hydrolysis rates in Jamesport sediments were 10- to 1,000-fold lower than rates previously reported for soils. In addition, aldicarb degradation rates were not significantly correlated with measurements of bacterial activity and density previously determined in the same sediments. Substantially higher aldicarb degradation rates were found in anoxic lake and salt marsh than in aquifer sediments. Furthermore, we investigated the anaerobic microbial processes involved in aldicarb transformation by adding organic substrates (acetate, glucose), an alternative electron acceptor (nitrate), and microbial inhibitors (molybdate, 2-bromoethanesulfonic acid) to anoxic aquifer, lake, and salt marsh sediments. The results suggest that a methanogenic consortium was important in aldicarb transformation or in the use of aldicarb-derived products such as methylamine. In addition, microbial aldicarb transformation proceeded via different pathways under oxic and anoxic conditions. In the presence of O(inf2), aldicarb transformation was mainly via an oxidation pathway, while in the absence of O(inf2), degradation took place through a hydrolytic pathway (including the formation of methylamine precursors). Under anoxic conditions, therefore, aldicarb can be transformed by microbial consortia to yield products which can be of direct benefit to natural populations of methanogens present in sediments.     

Internal wave-induced redox shifts affect biogeochemistry and microbial activity in sediments: a simulation experiment

Internal waves (seiches) are well-studied physical processes in stratified lakes, but their effects on sediment porewater chemistry and microbiology are still largely unexplored. Due to pycnocline oscillations, sediments are exposed to recurrent changes between epilimnetic and hypolimnetic water. This results in strong differences of environmental conditions, which should be reflected in the responses of redox-sensitive biogeochemical processes at both, the sediment–water interface and deeper sediment layers. We tested in a series of mesocosm experiments the influence of seiche-induced redox changes on porewater chemistry and bacterial activity in the sediments under well controlled conditions. Thereby, we excluded effects of changes in current and temperature regimes. For a period of 10 days, intact sediment cores from oligotrophic Lake Stechlin were incubated under constant (either oxic or anoxic) or alternating redox conditions. Solute concentrations were measured as porewater profiles in the sediment, while microbial activity was determined in the upper 0.5 cm of sediment. Oxic and alternating redox conditions resulted in similar ammonium, phosphate, and methane porewater concentrations, while concentrations of each analyte were considerably higher in anoxic cores. Microbial activity was clearly lower in the anoxic cores than in the oxic and the alternating cores. In conclusion, cores with intermittent anoxic phases of up to 24 hours do not differ in biogeochemistry and microbial activities from static oxic sediments. However, due to various physical processes seiches cause oxygen to penetrate deeper into sediment layers, which affects sediment redox gradients and increase microbial activity in seiche-influenced sediments.     

Effect of positive redox potentials (>+400mV) on the expression of anaerobic respiratory enzymes in Escherichia coli

The expression of fumarate reductase and other enzymes of anaerobic respiration in Escherichia coli was studied as a function of the redox potential (Eh) in the medium. Redox potentials up to +300 mV allowed full expression of fumarate reductase (frd) genes. Higher values resulted in decreased expression. The relationship between Eh and expression of frd could be approximated by the Nernst equation, assuming a redox couple with a midpoint potential Eo' = +400 mV to 440 mV. At Eh values greater than +510 mV (generated anaerobically by hexacyanoferrate(III] the degree of repression was the same as that obtained by O2. Hexacyanoferrate(III) also caused decreased activities of dimethylsulphoxide (DMSO), nitrite and nitrate reductases. Since expression of these enzymes depends on FNR, the gene activator of anaerobic respiratory genes, it is suggested that the function of FNR is controlled by a redox couple of Eo' = +400 mV to 440 mV.     

Hydrogen Concentration Profiles at the Oxic-Anoxic Interface: a Microsensor Study of the Hindgut of the Wood-Feeding Lower Termite Reticulitermes flavipes (Kollar)

Molecular hydrogen is a key intermediate in lignocellulose degradation by the microbial community of termite hindguts. With polarographic, Clark-type H(inf2) microelectrodes, we determined H(inf2) concentrations at microscale resolution in the gut of the wood-feeding lower termite Reticulitermes flavipes (Kollar). Axial H(inf2) concentration profiles obtained from isolated intestinal tracts embedded in agarose Ringer solution clearly identified the voluminous hindgut paunch as the site of H(inf2) production. The latter was strictly coupled with both a low redox potential (E(infh) = -200 mV) and the absence of oxygen, in agreement with the growth requirements of the cellulolytic, H(inf2)-producing flagellates located in the hindgut paunch. Luminal H(inf2) partial pressures were much higher than expected (ca. 5 kPa) and increased more than threefold when the guts were incubated under a N(inf2) headspace. Radial H(inf2) concentration gradients showed a steep decrease from the gut center towards the periphery, indicating the presence of H(inf2)-consuming activities both within the lumen and at the gut epithelium. Measurements under controlled gas headspace showed that the gut wall was also a sink for externally supplied H(inf2), both under oxic and anoxic conditions. With O(inf2) microelectrodes, we confirmed that the H(inf2) sink below the gut epithelium is located within the microoxic gut periphery, but the H(inf2)-consuming activity itself, at least a substantial part of it, was clearly due to an anaerobic process. These results are in accordance with the recently reported presence of methanogens attached in large numbers to the luminal side of the hindgut epithelium of R. flavipes. If the oxygen partial pressure was increased, O(inf2) penetrated deeper and H(inf2) production was suppressed; it ceased completely as soon as the gut was fully oxic. In experiments with living termites, externally supplied H(inf2) (20 kPa) stimulated methane formation five- to sixfold to 0.93 (mu)mol (g of termite)(sup-1) h(sup-1), indicating that the methanogenic activity in R. flavipes hindguts is not saturated for hydrogen under in situ conditions. This rate was in good agreement with the H(inf2) uptake rates exhibited by isolated hindguts, which would account for more than half of the CH(inf4) formed by living termites under comparable conditions.     

Differential regulation of periplasmic nitrate reductase gene (napKEFDABC) expression between aerobiosis and anaerobiosis with nitrate in a denitrifying phototroph Rhodobacter sphaeroides f. sp. denitrificans

A denitrifying phototroph, Rhodobacter sphaeroides f. sp. denitrificans, has the ability to denitrify by respiring nitrate. The periplasmic respiratory nitrate reductase (Nap) catalyses the first step in denitrification and is encoded by the genes, napKEFDABC. By assaying the ss-galactosidase activity of napKEFD-lacZ fusions in wild type and nap mutant cells grown under various growth conditions, the environmental signal for inducing nap expression was examined. Under anoxic conditions with nitrate, nap genes expression in the wild-type strain was highest in the dark, and somewhat lowered by incident light, but that of the napA, napB, and napC mutant strains was low, showing that nap expression is dependent on nitrate respiration. Under oxic conditions, both the wild type and nap mutant cells showed high ss-galactosidase activities, comparable to the wild-type grown under anoxic conditions with nitrate. Myxothiazol, a specific inhibitor of the cytochrome bc (1) complex, did not affect the beta-galactosidase activity in the wild-type cells grown aerobically, suggesting that the redox state of the quinone pool was not a candidate for the activation signal for aerobic nap expression. These results suggested that the trans-acting regulatory signals for nap expression differ between anoxic and oxic conditions. Deletion analysis showed that the nucleotide sequence from -135 to -88 with respect to the translational start point is essential for nap expression either under anoxic or oxic conditions, suggesting that the same cis-acting element is involved in regulating nap expression under either anoxic with nitrate or oxic conditions.     

Control of Methane Metabolism in a Forested Northern Wetland,New York State,by Aeration,Substrates, and Peat Size Fractions

Although many northern peat-forming wetlands (peatlands) are a suitable habitat for anaerobic CH 4 -producing bacteria (methanogens), net CH 4 fluxes are typically low in forested systems. We examined whether soil factors (aeration, substrate availability, peat size fractions) constrained net CH 4 production in peat from a Sphagnum -moss dominated, forested peatland in central New York State. The mean rate of net CH 4 production measured at 24° C was 79 nmol g -1 d -1 , and the mean rate of CO 2 production (respiration) was 5.7 w mol g -1 d -1 , in surface (0 to 10 cm) and subsurface (30 to 40 cm) peat. Saturated peat (900% water content) exposed to oxic conditions for 2 days or 14 days showed no net CH 4 production when subsequently exposed to anoxic conditions. Rates of CO 2 production, measured concomitantly, were essentially the same under oxic and anoxic conditions, and net CH 4 consumption under oxic conditions was barely affected by short-term exposure to anoxic conditions. Therefore, methanogens were particularly sensitive to aeration. Net CH 4 production in whole peat increased within hours of adding either acetate, glucose, or ethanol, substrates that methanogens can convert directly or indirectly into CH 4 , indicating that availability of these substrate might limit net CH 4 production in situ. In longer incubations of 30 days, only ethanol addition stimulated a large increase in net CH 4 production, suggesting growth in the population of methanogens when ethanol was available. We fractionated peat into size fractions and the largest sized fraction (> 1.19 mm), composed mostly of roots, showed the greatest net CH 4 production, although net CH 4 production in smaller fractions showed the largest response to ethanol addition. The circumstantial evidence presented here, that ethanol coming from plant roots supports net CH 4 production in forested sites, merits more research.     

Acetogenic Capacities and the Anaerobic Turnover of Carbon in a Kansas Prairie Soil

To assess the anaerobic capacities of a temperate grassland soil, a Kansas prairie soil was incubated anaerobically as either soil-water (1:2) suspensions or as soil microcosms at 78% soil water-holding capacity. Prairie soil formed acetate and CO(inf2) as the two main initial carbonaceous products from the anaerobic turnover of endogenous organic matter. Metabolic capacities of soil suspensions and microcosms were similar. Rates of acetate formation from endogenous organic matter in soil-water suspensions incubated at 40, 30, and 15(deg)C approximated 3.3, 2.4, and 1.1 (mu)g of acetate per g (dry weight) of soil per h, respectively. Supplemental H(inf2) and CO(inf2) were subject to consumption with the apparent concomitant synthesis of acetate in both soil suspensions and soil microcosms. In soil microcosms, rates of H(inf2)-dependent acetogenesis at 30 and 55(deg)C were nearly equivalent. The uptake of supplemental H(inf2) was not coupled to methanogenesis under any condition examined. These anaerobic activities were relatively stable when soils were subjected to either aerobic drying or alternating periods of O(inf2) enrichment. On the basis of the formation of nitrogen (N(inf2)), denitrification was engaged during anaerobic incubation periods; nitrous oxide (N(inf2)O) was also formed under certain conditions. Although extended incubation of soil induced the delayed methanogenic turnover of acetate, acetate was subject to immediate turnover under either O(inf2)- or nitrate-enriched conditions. These studies support the following concepts: (i) obligately anaerobic bacteria such as acetogenic bacteria are stable to periods of aerobiosis and are active in the anaerobic microsites of oxic soils, and (ii) acetate synthesized in anaerobic microsites of oxic terrestrial soils constitutes a trophic link to both aerobic and anaerobic microbial communities.     

Enterobacteriaceae facilitate the anaerobic degradation of glucose by a forest soil

Anoxic micro zones that occur in soil aggregates of oxic soils may be temporarily extended after rainfall and thus facilitate the anaerobic degradation of organic compounds in soils. The microbial degradation of glucose by anoxic slurries of a forest soil yielded acetate, CO₂, H₂, succinate, and ethanol, products indicative of mixed acid fermentation. Prokaryotes involved in this process were identified by time-resolved 16S rRNA gene-targeted stable isotope probing with [¹³C-U]-glucose. All labeled phylotypes from the ¹³C-enriched 16S rRNA gene were most closely related to Rahnella and Ewingella , enterobacterial genera known to catalyze mixed acid fermentation. These results indicate that facultative aerobes, in particular Enterobacteriaceae , (1) can outcompete obligate anaerobes when conditions become anoxic in forest soils and (2) may be involved in the initial decomposition of monosaccharides in anoxic micro zones of aerated forest soils.