53 research outputs found

    Solute Concentrations Influence Microbial Methanogenesis in Coal-bearing Strata of the Cherokee Basin, USA

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    Microorganisms have contributed significantly to subsurface energy resources by converting organic matter in hydrocarbon reservoirs into methane, the main component of natural gas. In this study, we consider environmental controls on microbial populations in coal-bearing strata of the Cherokee basin, an unconventional natural gas resource in southeast Kansas, USA. Pennsylvanian-age strata in the basin contain numerous thin (0.4-1.1 m) coalbeds with marginal thermal maturities (0.5-0.7% Ro) that are interbedded with shale and sandstone. We collected gas, water, and microbe samples from 16 commercial coalbed methane wells for geochemical and microbiological analysis. The water samples were Na-Cl type with total dissolved solids (TDS) content ranging from 34.9 to 91.3 g L?1. Gas dryness values [C1/(C2 + C3)] averaged 2640 and carbon and hydrogen isotope ratios of methane differed from those of carbon dioxide and water, respectively, by an average of 65 and 183%. These values are thought to be consistent with gas that formed primarily by hydrogenotrophic methanogenesis. Results from cultivation assays and taxonomic analysis of 16S rRNA genes agree with the geochemical results. Cultivable methanogens were present in every sample tested, methanogen sequences dominate the archaeal community in each sample (avg 91%), and few archaeal sequences (avg 4.2%) were classified within Methanosarcinales, an order of methanogens known to contain methylotrophic methanogens. Although hydrogenotrophs appear dominant, geochemical and microbial analyses both indicate that the proportion of methane generated by acetoclastic methanogens increases with the solute content of formation water, a trend that is contrary to existing conceptual models. Consistent with this trend, beta diversity analyses show that archaeal diversity significantly correlates with formation water solute content. In contrast, bacterial diversity more strongly correlates with location than solute content, possibly as a result of spatial variation in the thermal maturity of the coalbeds.Citation: Kirk MF, Wilson BH, Marquart KA, Zeglin LH, Vinson DS and Flynn TM (2015) Solute Concentrations Influence Microbial Methanogenesis in Coal-bearing Strata of the Cherokee Basin, USA. Front. Microbiol. 6:1287. doi: 10.3389/fmicb.2015.0128

    Innate and adaptive humoral responses coat distinct commensal bacteria with immunoglobulin A

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    Immunoglobulin A (IgA) is prominently secreted at mucosal surfaces and coats a fraction of the intestinal microbiota. However, the commensal bacteria bound by IgA are poorly characterized and the type of humoral immunity they elicit remains elusive. We used bacterial flow cytometry coupled with 16S rRNA gene sequencing (IgA-Seq) in murine models of immunodeficiency to identify IgA-bound bacteria and elucidate mechanisms of commensal IgA targeting. We found that residence in the small intestine, rather than bacterial identity, dictated induction of specific IgA. Most commensals elicited strong T-independent (TI) responses that originated from the orphan B1b lineage and from B2 cells, but excluded natural antibacterial B1a specificities. Atypical commensals including segmented filamentous bacteria and Mucispirillum evaded TI responses but elicited T-dependent IgA. These data demonstrate exquisite targeting of distinct commensal bacteria by multiple layers of humoral immunity and reveal a specialized function of the B1b lineage in TI mucosal IgA responses

    Microbial diversity and groundwater chemistry in a pristine aquifer

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    The Mahomet aquifer, a pristine water resource spanning much of central Illinois, is home to a diverse ecosystem of bacteria and archaea. We examined this microbial community at 18 wells across the aquifer by filtering cells suspended in groundwater and trapping sediment-bound populations on sterile samplers incubated in the wells themselves. After extracting DNA directly from these samples, we characterized both the suspended and attached communities with terminal restriction fragment length polymorphism (T-RFLP) and large-scale sequencing of the 16S rRNA gene. Using multivariate statistics, we quantified how the distribution of bacterial populations corresponds to the geochemical zonation of groundwater. Groundwater in this area of the Mahomet aquifer is divided into areas of high and low sulfate, and the concentration of dissolved methane in these areas is inversely proportional to that of sulfate. Although groundwater in every well we sampled was considerably richer in ferrous iron than sulfide, we found that the presence of iron-reducing bacteria does not exclude sulfate reducers as previously assumed. Instead, where the concentration of sulfate is high, sulfate reducers comprise a proportion of the bacterial community (18%) nearly equivalent to that of iron reducers (23%). Iron reducers only dominate the bacterial community in wells with sulfate concentrations of 0.2 mM or less, where clones classified as Geobacter, Geothrix, or Desulfuromonas represent more than half of all sequences analyzed. We observed, in contrast, no statistically significant relationship between the structure of the bacterial community and the concentration of either ferrous iron or dissolved sulfide. This result calls into question the use of iron and sulfide as indicators of the nature of subsurface bacterial activity. Sulfate reducers and iron reducers do not appear to be segregated into discrete zones in the aquifer, as commonly believed to occur as a result of competitive exclusion. Instead, we found the two groups co-existing in the subsurface in a relationship that we argue is not only competitive, but mutualistic. Through further use of sediment traps to evaluate the attached microbial community, we found that overall structure of bacterial communities in the Mahomet is resistant to a shift in the availability of sulfate. After one year of incubation within a well where the concentration of sulfate was either high (1.5 mM) or low (0.04 mM), sediment traps were switched between the two wells for an additional year of incubation. Despite the more than 40-fold change in the concentration of sulfate, these switched traps remained more similar to the community from the well in which they were initially incubated. While the overall community composition did not change significantly, certain bacterial groups associated with sulfate reduction (Desulfobacter and Desulfobulbus) were found to increase or decrease along with the concentration of sulfate. These results show that while the abundance of many populations is at least partly controlled by the evolutionary history of that particular community, certain functional groups of critical biogeochemical importance are sensitive to local changes. While measures of overall community similarity are useful, the specific abundance of these taxa must be accounted for

    Electron Donor Utilization and Secondary Mineral Formation during the Bioreduction of Lepidocrocite by Shewanella putrefaciens CN32

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    The bioreduction of Fe(III) oxides by dissimilatory iron reducing bacteria (DIRB) may result in the production of a suite of Fe(II)-bearing secondary minerals, including magnetite, siderite, vivianite, green rusts, and chukanovite; the formation of specific phases controlled by the interaction of various physiological and geochemical factors. In an effort to better understand the effects of individual electron donors on the formation of specific Fe(II)-bearing secondary minerals, we examined the effects of a series of potential electron donors on the bioreduction of lepidocrocite (γ-FeOOH) by Shewanella putrefaciens CN32. Biomineralization products were identified by X-ray diffraction, Mössbauer spectroscopy, and scanning electron microscopy. Acetate, citrate, ethanol, glucose, glutamate, glycerol, malate, and succinate were not effectively utilized for the bioreduction of lepidocrocite by S. putrefaciens CN32; however, substantial Fe(II) production was observed when formate, lactate, H2, pyruvate, serine, or N acetylglucosamine (NAG) was provided as an electron donor. Carbonate or sulfate green rust was the dominant Fe(II)-bearing secondary mineral when formate, H2, lactate, or NAG was provided, however, siderite formed with pyruvate or serine. Geochemical modeling indicated that pH and carbonate concentration are the key factors determining the prevalence of carbonate green rust verses siderite

    Parallelized, Aerobic, Single Carbon-Source Enrichments from Different Natural Environments Contain Divergent Microbial Communities

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    Microbial communities that inhabit environments such as soil can contain thousands of distinct taxa, yet little is known about how this diversity is maintained in response to environmental perturbations such as changes in the availability of carbon. By utilizing aerobic substrate arrays to examine the effect of carbon amendment on microbial communities taken from six distinct environments (soil from a temperate prairie and forest, tropical forest soil, subalpine forest soil, and surface water and soil from a palustrine emergent wetland), we examined how carbon amendment and inoculum source shape the composition of the community in each enrichment. Dilute subsamples from each environment were used to inoculate 96-well microtiter plates containing triplicate wells amended with one of 31 carbon sources from six different classes of organic compounds (phenols, polymers, carbohydrates, carboxylic acids, amines, amino acids). After incubating each well aerobically in the dark for 72 h, we analyzed the composition of the microbial communities on the substrate arrays as well as the initial inocula by sequencing 16S rRNA gene amplicons using the Illumina MiSeq platform. Comparisons of alpha and beta diversity in these systems showed that, while the composition of the communities that grow to inhabit the wells in each substrate array diverges sharply from that of the original community in the inoculum, these enrichment communities are still strongly affected by the inoculum source. We found most enrichments were dominated by one or several OTUs most closely related to aerobes or facultative anaerobes from the Proteobacteria (e.g., Pseudomonas, Burkholderia, and Ralstonia) or Bacteroidetes (e.g., Chryseobacterium). Comparisons within each substrate array based on the class of carbon source further show that the communities inhabiting wells amended with a carbohydrate differ significantly from those enriched with a phenolic compound. Selection therefore seems to play a role in shaping the communities in the substrate arrays, although some stochasticity is also seen whereby several replicate wells within a single substrate array display strongly divergent community compositions. Overall, the use of highly parallel substrate arrays offers a promising path forward to study the response of microbial communities to perturbations in a changing environment
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