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Sample records for including cultivated methanogens

  1. Cultivation of methanogenic community from subseafloor sediments using a continuous-flow bioreactor

    PubMed Central

    Imachi, Hiroyuki; Aoi, Ken; Tasumi, Eiji; Saito, Yumi; Yamanaka, Yuko; Saito, Yayoi; Yamaguchi, Takashi; Tomaru, Hitoshi; Takeuchi, Rika; Morono, Yuki; Inagaki, Fumio; Takai, Ken

    2011-01-01

    Microbial methanogenesis in subseafloor sediments is a key process in the carbon cycle on the Earth. However, the cultivation-dependent evidences have been poorly demonstrated. Here we report the cultivation of a methanogenic microbial consortium from subseafloor sediments using a continuous-flow-type bioreactor with polyurethane sponges as microbial habitats, called down-flow hanging sponge (DHS) reactor. We anaerobically incubated methane-rich core sediments collected from off Shimokita Peninsula, Japan, for 826 days in the reactor at 10 °C. Synthetic seawater supplemented with glucose, yeast extract, acetate and propionate as potential energy sources was provided into the reactor. After 289 days of operation, microbiological methane production became evident. Fluorescence in situ hybridization analysis revealed the presence of metabolically active microbial cells with various morphologies in the reactor. DNA- and RNA-based phylogenetic analyses targeting 16S rRNA indicated the successful growth of phylogenetically diverse microbial components during cultivation in the reactor. Most of the phylotypes in the reactor, once it made methane, were more closely related to culture sequences than to the subsurface environmental sequence. Potentially methanogenic phylotypes related to the genera Methanobacterium, Methanococcoides and Methanosarcina were predominantly detected concomitantly with methane production, while uncultured archaeal phylotypes were also detected. Using the methanogenic community enrichment as subsequent inocula, traditional batch-type cultivations led to the successful isolation of several anaerobic microbes including those methanogens. Our results substantiate that the DHS bioreactor is a useful system for the enrichment of numerous fastidious microbes from subseafloor sediments and will enable the physiological and ecological characterization of pure cultures of previously uncultivated subseafloor microbial life. PMID:21654849

  2. Cultivation of methanogenic community from 2-km deep subseafloor coalbeds using a continuous-flow bioreactor

    NASA Astrophysics Data System (ADS)

    Imachi, H.; Tasumi, E.; Morono, Y.; Ito, M.; Takai, K.; Inagaki, F.

    2013-12-01

    Deep subseafloor environments associated with hydrocarbon reservoirs have been least explored by previous scientific drilling and hence the nature of deep subseafloor life and its ecological roles in the carbon cycle remain largely unknown. In this study, we performed cultivation of subseafloor methanogenic communities using a continuous-flow bioreactor with polyurethane sponges, called down-flow hanging sponge (DHS) reactor. The sample used for the reactor cultivation was obtained from 2 km-deep coalbeds off the Shimokita Peninsula of Japan, the northwestern Pacific, during the Integrated Ocean Drilling Program (IODP) Expedition 337 using a riser drilling technology of the drilling vessel Chikyu. The coalbed samples were incubated anaerobically in the DHS reactor at the in-situ temperature of 40°C. Synthetic seawater supplemented with a tiny amount of yeast extract, acetate, propionate and butyrate was provided into the DHS reactor. After 34 days of the bioreactor operation, a small production of methane was observed. The methane concentration was gradually increased and the stable carbon isotopic composition of methane was consistency 13C-depleted during the bioreactor operation, indicating the occurrence of microbial methanogenesis. Microscopic observation showed that the enrichment culture contained a variety of microorganisms, including methanogen-like rod-shaped cells with F420 auto-fluorescence. Interestingly, many spore-like particles were observed in the bioreactor enrichment. Phylogenetic analysis of 16S rRNA genes showed the growth of phylogenetically diverse bacteria and archaea in the DHS reactor. Predominant archaeal components were closely related to hydrogenotrophic methanogens within the genus Methanobacterium. Some predominant bacteria were related to the spore-formers within the class Clostridia, which are overall in good agreement with microscopic observations. By analyzing ion images using a nano-scale secondary ion mass spectrometry (Nano

  3. Genome sequence of a mesophilic hydrogenotrophic methanogen Methanocella paludicola, the first cultivated representative of the order Methanocellales.

    PubMed

    Sakai, Sanae; Takaki, Yoshihiro; Shimamura, Shigeru; Sekine, Mitsuo; Tajima, Takahisa; Kosugi, Hiroki; Ichikawa, Natsuko; Tasumi, Eiji; Hiraki, Aiko T; Shimizu, Ai; Kato, Yumiko; Nishiko, Rika; Mori, Koji; Fujita, Nobuyuki; Imachi, Hiroyuki; Takai, Ken

    2011-01-01

    We report complete genome sequence of a mesophilic hydrogenotrophic methanogen Methanocella paludicola, the first cultured representative of the order Methanocellales once recognized as an uncultured key archaeal group for methane emission in rice fields. The genome sequence of M. paludicola consists of a single circular chromosome of 2,957,635 bp containing 3004 protein-coding sequences (CDS). Genes for most of the functions known in the methanogenic archaea were identified, e.g. a full complement of hydrogenases and methanogenesis enzymes. The mixotrophic growth of M. paludicola was clarified by the genomic characterization and re-examined by the subsequent growth experiments. Comparative genome analysis with the previously reported genome sequence of RC-I(MRE50), which was metagenomically reconstructed, demonstrated that about 70% of M. paludicola CDSs were genetically related with RC-I(MRE50) CDSs. These CDSs included the genes involved in hydrogenotrophic methane production, incomplete TCA cycle, assimilatory sulfate reduction and so on. However, the genetic components for the carbon and nitrogen fixation and antioxidant system were different between the two Methanocellales genomes. The difference is likely associated with the physiological variability between M. paludicola and RC-I(MRE50), further suggesting the genomic and physiological diversity of the Methanocellales methanogens. Comparative genome analysis among the previously determined methanogen genomes points to the genome-wide relatedness of the Methanocellales methanogens to the orders Methanosarcinales and Methanomicrobiales methanogens in terms of the genetic repertoire. Meanwhile, the unique evolutionary history of the Methanocellales methanogens is also traced in an aspect by the comparative genome analysis among the methanogens. PMID:21829548

  4. Genome Sequence of a Mesophilic Hydrogenotrophic Methanogen Methanocella paludicola, the First Cultivated Representative of the Order Methanocellales

    PubMed Central

    Sakai, Sanae; Takaki, Yoshihiro; Shimamura, Shigeru; Sekine, Mitsuo; Tajima, Takahisa; Kosugi, Hiroki; Ichikawa, Natsuko; Tasumi, Eiji; Hiraki, Aiko T.; Shimizu, Ai; Kato, Yumiko; Nishiko, Rika; Mori, Koji; Fujita, Nobuyuki; Imachi, Hiroyuki; Takai, Ken

    2011-01-01

    We report complete genome sequence of a mesophilic hydrogenotrophic methanogen Methanocella paludicola, the first cultured representative of the order Methanocellales once recognized as an uncultured key archaeal group for methane emission in rice fields. The genome sequence of M. paludicola consists of a single circular chromosome of 2,957,635 bp containing 3004 protein-coding sequences (CDS). Genes for most of the functions known in the methanogenic archaea were identified, e.g. a full complement of hydrogenases and methanogenesis enzymes. The mixotrophic growth of M. paludicola was clarified by the genomic characterization and re-examined by the subsequent growth experiments. Comparative genome analysis with the previously reported genome sequence of RC-IMRE50, which was metagenomically reconstructed, demonstrated that about 70% of M. paludicola CDSs were genetically related with RC-IMRE50 CDSs. These CDSs included the genes involved in hydrogenotrophic methane production, incomplete TCA cycle, assimilatory sulfate reduction and so on. However, the genetic components for the carbon and nitrogen fixation and antioxidant system were different between the two Methanocellales genomes. The difference is likely associated with the physiological variability between M. paludicola and RC-IMRE50, further suggesting the genomic and physiological diversity of the Methanocellales methanogens. Comparative genome analysis among the previously determined methanogen genomes points to the genome-wide relatedness of the Methanocellales methanogens to the orders Methanosarcinales and Methanomicrobiales methanogens in terms of the genetic repertoire. Meanwhile, the unique evolutionary history of the Methanocellales methanogens is also traced in an aspect by the comparative genome analysis among the methanogens. PMID:21829548

  5. 29 CFR 780.110 - Operations included in “cultivation and tillage of the soil.”

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... 29 Labor 3 2012-07-01 2012-07-01 false Operations included in âcultivation and tillage of the soil... FAIR LABOR STANDARDS ACT General Scope of Agriculture Cultivation and Tillage of the Soil § 780.110 Operations included in “cultivation and tillage of the soil.” “Cultivation and tillage of the soil”...

  6. 29 CFR 780.110 - Operations included in “cultivation and tillage of the soil.”

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... 29 Labor 3 2013-07-01 2013-07-01 false Operations included in âcultivation and tillage of the soil... FAIR LABOR STANDARDS ACT General Scope of Agriculture Cultivation and Tillage of the Soil § 780.110 Operations included in “cultivation and tillage of the soil.” “Cultivation and tillage of the soil”...

  7. 29 CFR 780.110 - Operations included in “cultivation and tillage of the soil.”

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... 29 Labor 3 2014-07-01 2014-07-01 false Operations included in âcultivation and tillage of the soil... FAIR LABOR STANDARDS ACT General Scope of Agriculture Cultivation and Tillage of the Soil § 780.110 Operations included in “cultivation and tillage of the soil.” “Cultivation and tillage of the soil”...

  8. Novel molecular markers for the detection of methanogens and phylogenetic analyses of methanogenic communities.

    PubMed

    Dziewit, Lukasz; Pyzik, Adam; Romaniuk, Krzysztof; Sobczak, Adam; Szczesny, Pawel; Lipinski, Leszek; Bartosik, Dariusz; Drewniak, Lukasz

    2015-01-01

    Methanogenic Archaea produce approximately one billion tons of methane annually, but their biology remains largely unknown. This is partially due to the large phylogenetic and phenotypic diversity of this group of organisms, which inhabit various anoxic environments including peatlands, freshwater sediments, landfills, anaerobic digesters and the intestinal tracts of ruminants. Research is also hampered by the inability to cultivate methanogenic Archaea. Therefore, biodiversity studies have relied on the use of 16S rRNA and mcrA [encoding the α subunit of the methyl coenzyme M (methyl-CoM) reductase] genes as molecular markers for the detection and phylogenetic analysis of methanogens. Here, we describe four novel molecular markers that should prove useful in the detailed analysis of methanogenic consortia, with a special focus on methylotrophic methanogens. We have developed and validated sets of degenerate PCR primers for the amplification of genes encoding key enzymes involved in methanogenesis: mcrB and mcrG (encoding β and γ subunits of the methyl-CoM reductase, involved in the conversion of methyl-CoM to methane), mtaB (encoding methanol-5-hydroxybenzimidazolylcobamide Co-methyltransferase, catalyzing the conversion of methanol to methyl-CoM) and mtbA (encoding methylated [methylamine-specific corrinoid protein]:coenzyme M methyltransferase, involved in the conversion of mono-, di- and trimethylamine into methyl-CoM). The sensitivity of these primers was verified by high-throughput sequencing of PCR products amplified from DNA isolated from microorganisms present in anaerobic digesters. The selectivity of the markers was analyzed using phylogenetic methods. Our results indicate that the selected markers and the PCR primer sets can be used as specific tools for in-depth diversity analyses of methanogenic consortia. PMID:26217325

  9. Novel molecular markers for the detection of methanogens and phylogenetic analyses of methanogenic communities

    PubMed Central

    Dziewit, Lukasz; Pyzik, Adam; Romaniuk, Krzysztof; Sobczak, Adam; Szczesny, Pawel; Lipinski, Leszek; Bartosik, Dariusz; Drewniak, Lukasz

    2015-01-01

    Methanogenic Archaea produce approximately one billion tons of methane annually, but their biology remains largely unknown. This is partially due to the large phylogenetic and phenotypic diversity of this group of organisms, which inhabit various anoxic environments including peatlands, freshwater sediments, landfills, anaerobic digesters and the intestinal tracts of ruminants. Research is also hampered by the inability to cultivate methanogenic Archaea. Therefore, biodiversity studies have relied on the use of 16S rRNA and mcrA [encoding the α subunit of the methyl coenzyme M (methyl-CoM) reductase] genes as molecular markers for the detection and phylogenetic analysis of methanogens. Here, we describe four novel molecular markers that should prove useful in the detailed analysis of methanogenic consortia, with a special focus on methylotrophic methanogens. We have developed and validated sets of degenerate PCR primers for the amplification of genes encoding key enzymes involved in methanogenesis: mcrB and mcrG (encoding β and γ subunits of the methyl-CoM reductase, involved in the conversion of methyl-CoM to methane), mtaB (encoding methanol-5-hydroxybenzimidazolylcobamide Co-methyltransferase, catalyzing the conversion of methanol to methyl-CoM) and mtbA (encoding methylated [methylamine-specific corrinoid protein]:coenzyme M methyltransferase, involved in the conversion of mono-, di- and trimethylamine into methyl-CoM). The sensitivity of these primers was verified by high-throughput sequencing of PCR products amplified from DNA isolated from microorganisms present in anaerobic digesters. The selectivity of the markers was analyzed using phylogenetic methods. Our results indicate that the selected markers and the PCR primer sets can be used as specific tools for in-depth diversity analyses of methanogenic consortia. PMID:26217325

  10. Higher Temperature and Hydrogen Availability Stimulated the Methanogenic Activity in East Antarctic Subglacial Sediment

    NASA Astrophysics Data System (ADS)

    Ma, H.

    2014-12-01

    Subglacial ecosystem has been recognized as an environment with considerable methanogenic activity, and therefore is of significant impact on global methane budget and climate change. Although the methanogens have been discovered at a few subglacial environments, the methanogenic activity there is yet insufficiently studied, especially on the effects of environmental parameters, due to technical difficulties on sampling and cultivation. Here, in this study, we attempt to access the methanogenic activity and community structure in response to temperature and substrate availability. An integrated approach including in vitro cultivation and molecular techniques were employed. A subglacial sediment from Larsemann Hills, East Antarctica was incubated at different temperatures (1, 4, 12 oC) supplied with H2+CO2 or sodium acetate to estimate the methanogenic activity. The McrA gene which is a specific marker for methanogens was amplified with primer ME and ML to construct phylogenetic trees. This functional gene was also quantified by Q-PCR before and after the incubation to estimate the increase of methanogens. After 8 months a highest methanogenesis rate of 226 pmol/ day/ gram sediment was observed at 12 oC with H2 supplying, which was 2 times higher than that with acetate supplying, clearly suggesting that H2 is a preferable substrate than acetate. The methanogenesis rate without supplying extra substrate showed positive temperature dependence with rate of 23.3, 24.8, 131 pmol/day/gram sediment at 1 oC, 4 oC, and 12 oC, respectively. The McrA copy number was increased more than 300 times and 50 times with H2 and acetate supplying respectively after the incubation. 94% and 67% of the mcrA gene sequences were classed into methanomicrobiales which were hydrogen-trophic methanogens in the two clone libraries with primer ML and ME respectively. This finding suggests the potential effect of methanogenesis under glacier on the climate change.

  11. 29 CFR 780.110 - Operations included in “cultivation and tillage of the soil.”

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... the ground for a proper seedbed or building terraces on farmland to check soil erosion are included... 29 Labor 3 2010-07-01 2010-07-01 false Operations included in âcultivation and tillage of the soil... FAIR LABOR STANDARDS ACT General Scope of Agriculture Cultivation and Tillage of the Soil §...

  12. New approach to the cultivation of methanogenic bacteria: 2-mercaptoethanesulfonic acid (HS-CoM)-dependent growth of Methanobacterium ruminantium in a pressureized atmosphere.

    PubMed Central

    Balch, W E; Wolfe, R S

    1976-01-01

    The sensitivity of the requirement of Methanobacterium ruminantium strain M1 to a new coenzyme, 2-mercaptoethanesulfonic acid (HS-CoM) was examined by use of new techniques that were developed for rapid and efficient handling of large numbers of cultures of methanogenic bacteria. The system uses sealed tubes that contain a gas mixture of 80% hydrogen and 20% carbon dioxide under a pressure of 2 to 3 atm. This modification of the Hungate technique reduces variability among replicate cultures and simplifies the dispensing, sterilization, and storage of liquid media as well as the transfer and maintenance of methanogenic bacteria. Results indicate a limit of sensitivity of the assay at 5 nM HS-CoM, with half-maximal growth at 25 nM HS-CoM. Coenzyme activity could be replaced by 2,2'-dithiodiethanesulfonic acid at a half-molar equivalent of the HS-CoM concentration, or by 2-(methylthio)ethanesulfonic acid on an equimolar basis. These data reveal a very sensitive and precise requirement for HS-CoM in the nutrition of this fastidious anaerobe. Images PMID:827241

  13. Phylogenetic Characterization of Methanogenic Assemblages in Eutrophic and Oligotrophic Areas of the Florida Everglades†

    PubMed Central

    Castro, Hector; Ogram, Andrew; Reddy, K. R.

    2004-01-01

    Agricultural activities have produced well-documented changes in the Florida Everglades, including establishment of a gradient in phosphorus concentrations in Water Conservation Area 2A (WCA-2A) of the northern Everglades. An effect of increased phosphorus concentrations is increased methanogenesis in the eutrophic regions compared to the oligotrophic regions of WCA-2A. The goal of this study was to identify relationships between eutrophication and composition and activity of methanogenic assemblages in WCA-2A soils. Distributions of two genes associated with methanogens were characterized in soils taken from WCA-2A: the archaeal 16S rRNA gene and the methyl coenzyme M reductase gene. The richness of methanogen phylotypes was greater in eutrophic than in oligotrophic sites, and sequences related to previously cultivated and uncultivated methanogens were found. A preferential selection for the order Methanomicrobiales was observed in mcrA clone libraries, suggesting primer bias for this group. A greater diversity within the Methanomicrobiales was observed in mcrA clone libraries than in 16S rRNA gene libraries. 16S rRNA phylogenetic analyses revealed a dominance of clones related to Methanosaeta spp., an acetoclastic methanogen dominant in environments with low acetate concentrations. A significant number of clones were related to Methanomicrobiales, an order characterized by species utilizing hydrogen and formate as methanogenic substrates. No representatives of the orders Methanobacteriales and Methanococcales were found in any 16S rRNA clone library, although some Methanobacteriales were found in mcrA libraries. Hydrogenotrophs are the dominant methanogens in WCA-2A, and acetoclastic methanogen genotypes that proliferate in low acetate concentrations outnumber those that typically dominate in higher acetate concentrations. PMID:15528519

  14. The Geobiochemistry of Methanogen Proteins

    NASA Astrophysics Data System (ADS)

    Prasad, A.; Shock, E.

    2013-12-01

    A principle of geobiochemistry is that adaptation over evolutionary time includes a thermodynamic drive to minimize costs of making biomolecules like proteins and lipids. If so, then biomolecule abundances will reflect, at least in part, their relative stabilities at the conditions imposed by external environments. We tested this hypothesis by comparing relative stabilities of 138 orthologous proteins between a representative lake-sediment methanogen (Methanoculleus marisnigri) and a representative rumen methanogen (Methanospirillum hungatei) at the compositional constraints of their respective environments. Chemical affinities of the proteins were calculated based on pH, temperature, and concentrations of dissolved hydrogen, bicarbonate, ammonia, and hydrogen sulfide, together with standard Gibbs energies of formation of proteins from the elements predicted with a group additivity algorithm for unfolded proteins [1]. Methanogens were chosen as they are chemoautotrophs and their metabolism proceeds at relatively small affinities. Also, they are found in a variety of compositionally varying habitats like rumen, sediments, hydrothermal systems and sewage. The methanogens selected belong to the same order of taxonomy and are closely related. Preliminary results show that a majority of the proteins belonging to the rumen methanogen (66%) are more stable in the rumen environment, while a majority of the proteins belonging to the lake-sediment methanogen (58%) are more stable at sediment conditions. In a separate observation, it was noted that while the complete protein ';proteasome subunit alpha' of another rumen methanogen (Methanobrevibacter smithii) was less stable in its more reducing habitat as compared to a sewage methanogen (Methanothermobacter thermoautotophicus), its first 26 amino acid residues (N terminal) were in fact more stable in its own environment. These 26 residues are reported to be unique as compared to other proteasome proteins and are suggested to

  15. Methanogens in the Solar System

    NASA Astrophysics Data System (ADS)

    Taubner, Ruth-Sophie; Schleper, Christa; Firneis, Maria G.; Rittmann, Simon

    2015-04-01

    The last decade of space science revealed that potential habitats in the Solar System may not be limited to the classical habitable zone supporting life as we know it. These microorganisms were shown to thrive under extremophilic growth conditions. Here, we outline the main eco-physiological characteristics of methanogens like their response on temperature, pressure, or pH changes or their resistance against radiation or desiccation. They can withstand extreme environmental conditions which makes them intriguing organisms for astrobiological studies. On Earth, they are found for example in wetlands, in arctic and antarctic subglacial environments, in ruminants, and even in the environment surrounding the Mars Desert Research Station in Utah. These obligate anaerobic chemolithoautotrophs or chemolithoheterotrophs are able to use e.g. hydrogen and C1 compounds like CO2, formate, or methanol as energy source and carbon source, respectively. We point out their capability to be able to habitat potential extraterrestrial biospheres all over the planetary system. We will give an overview about these possible environments on Mars, icy moons like Europa or Enceladus, and minor planets. We present an overview about studies of methanogens with an astrobiological relevance and we show our conclusions about the role of methanogens for the search for extraterrestrial life in the Solar System. We will present first results of our study about the possibility to cultivate methanogens under Enceladus-like conditions. For that, based on the observations obtained by the Cassini spacecraft concerning the plume compounds, we produce a medium with a composition similar to the ocean composition of this icy moon which is far more Enceladus-like than in any (published) experiment before. Eventually, we give an outlook on the feasibility and the necessity of future astrobiological studies with these microbes. We point out the importance of future in-situ or even sample and return missions to

  16. Greenhouse gas emissions from shifting cultivation in the tropics, including uncertainty and sensitivity analysis

    NASA Astrophysics Data System (ADS)

    Silva, J. M. N.; Carreiras, J. M. B.; Rosa, I.; Pereira, J. M. C.

    2011-10-01

    Annual emissions of CO2, CH4, CO, N2O, and NOx from biomass burning in shifting cultivation systems in tropical Asia, Africa, and America were estimated at national and continental levels as the product of area burned, aboveground biomass, combustion completeness, and emission factor. The total area of shifting cultivation in each country was derived from the Global Land Cover 2000 map, while the area cleared and burned annually was obtained by multiplying the total area by the rotation cycle of shifting cultivation, calculated using cropping and fallow lengths reported in the literature. Aboveground biomass accumulation was estimated as a function of the duration and mean temperature of the growing season, soil texture type, and length of the fallow period. The uncertainty associated with each model variable was estimated, and an uncertainty and sensitivity analysis of greenhouse gas estimates was performed with Monte Carlo and variance decomposition techniques. Our results reveal large uncertainty in emission estimates for all five gases. In the case of CO2, mean (standard deviation) emissions from shifting cultivation in Asia, Africa, and America were estimated at 241 (132), 205 (139), and 295 (197) Tg yr-1, respectively. Combustion completeness and emission factors were the model inputs that contributed the most to the uncertainty of estimates. Our mean estimates are lower than the literature values for atmospheric emission from biomass burning in shifting cultivation systems. Only mean values could be compared since other studies do not provide any measure of uncertainty.

  17. Hydrogenotrophic methanogens dominate in biogas reactors fed with defined substrates.

    PubMed

    Kampmann, K; Ratering, S; Baumann, R; Schmidt, M; Zerr, W; Schnell, S

    2012-09-01

    Methanogenic communities in 200L biogas reactors containing liquid manure were investigated for 33 d. The reactors were consecutively fed with casein, starch and cream. Real-time PCR with primers targeting the gene for methyl coenzyme-M reductase (mcrA) resulted in copy numbers of up to 2.1×10(9) g dry mass(-1). Single strand conformation polymorphism (SSCP) analysis revealed a stable community consisting of few hydrogenotrophic methanogens. One of the two most abundant species was closely related to Methanospirillum hungatei, whereas the other one was only distantly related to other methanogens, with Methanopyrus kandleri being the closest cultivated relative. Most probable number (MPN) cultivations were accomplished with a sample from a 600 m(3) reactor from which all manures used in the experiments originated, and equal cell counts of ca. 10(9) g dry mass(-1) were found for cultivations with acetate, H(2) and methanol. SSCP analysis of these samples and sequencing of the DNA bands identified different hydrogenotrophic methanogens in all samples, and acetoclastic methanogens closely related to Methanosarcina mazei in the samples cultivated with acetate and methanol. As the acetoclastic species were not found in any other SSCP sample, it was supposed that the ammonia values in the manure of the laboratory biogas reactor, which ranged from 2.48 to 3.61 g NH(4)-NL(-1), inhibited the growth of the acetoclastic methanogens. PMID:22918024

  18. 29 CFR 780.110 - Operations included in “cultivation and tillage of the soil.”

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... 29 Labor 3 2011-07-01 2011-07-01 false Operations included in âcultivation and tillage of the soil.â 780.110 Section 780.110 Labor Regulations Relating to Labor (Continued) WAGE AND HOUR DIVISION, DEPARTMENT OF LABOR STATEMENTS OF GENERAL POLICY OR INTERPRETATION NOT DIRECTLY RELATED TO REGULATIONS EXEMPTIONS APPLICABLE TO AGRICULTURE,...

  19. Osmoregulation in methanogens

    SciTech Connect

    Roberts, M.F.

    1993-01-01

    Our major goal of our work has been to develop and use NMR techniques to study how methanogenic archaebacteria deal with osmotic stress with the hope of providing insights into increasing the salt tolerance of other cells. The project has three main sections: (i) in vivo studies of methanogens; (ii) use of [sup l3]C- and [sup l5]N- labeled potential precursors and in vitro analyses of specific label uptake for elucidation of osmolyte dynamics and biosynthetic pathways of osmolytes in these organisms, and isolation of key biosynthetic enzymes; and (iii) collaborative studies on identification of organic solutes in other methanogens.

  20. Microbial dark matter ecogenomics reveals complex synergistic networks in a methanogenic bioreactor.

    PubMed

    Nobu, Masaru K; Narihiro, Takashi; Rinke, Christian; Kamagata, Yoichi; Tringe, Susannah G; Woyke, Tanja; Liu, Wen-Tso

    2015-08-01

    Ecogenomic investigation of a methanogenic bioreactor degrading terephthalate (TA) allowed elucidation of complex synergistic networks of uncultivated microorganisms, including those from candidate phyla with no cultivated representatives. Our previous metagenomic investigation proposed that Pelotomaculum and methanogens may interact with uncultivated organisms to degrade TA; however, many members of the community remained unaddressed because of past technological limitations. In further pursuit, this study employed state-of-the-art omics tools to generate draft genomes and transcriptomes for uncultivated organisms spanning 15 phyla and reports the first genomic insight into candidate phyla Atribacteria, Hydrogenedentes and Marinimicrobia in methanogenic environments. Metabolic reconstruction revealed that these organisms perform fermentative, syntrophic and acetogenic catabolism facilitated by energy conservation revolving around H2 metabolism. Several of these organisms could degrade TA catabolism by-products (acetate, butyrate and H2) and syntrophically support Pelotomaculum. Other taxa could scavenge anabolic products (protein and lipids) presumably derived from detrital biomass produced by the TA-degrading community. The protein scavengers expressed complementary metabolic pathways indicating syntrophic and fermentative step-wise protein degradation through amino acids, branched-chain fatty acids and propionate. Thus, the uncultivated organisms may interact to form an intricate syntrophy-supported food web with Pelotomaculum and methanogens to metabolize catabolic by-products and detritus, whereby facilitating holistic TA mineralization to CO2 and CH4. PMID:25615435

  1. Ammonia effect on hydrogenotrophic methanogens and syntrophic acetate-oxidizing bacteria.

    PubMed

    Wang, Han; Fotidis, Ioannis A; Angelidaki, Irini

    2015-11-01

    Ammonia-rich substrates can cause inhibition on anaerobic digestion process. Syntrophic acetate-oxidizing bacteria (SAOB) and hydrogenotrophic methanogens are important for the ammonia inhibitory mechanism on anaerobic digestion. The roles and interactions of SAOB and hydrogenotrophic methanogens to ammonia inhibition effect are still unclear. The aim of the current study was to determine the ammonia toxicity levels of various pure strains of SAOB and hydrogenotrophic methanogens. Moreover, ammonia toxicity on the syntrophic-cultivated strains of SAOB and hydrogenotrophic methanogens was tested. Thus, four hydrogenotrophic methanogens (i.e. Methanoculleus bourgensis, Methanobacterium congolense, Methanoculleu thermophilus and Methanothermobacter thermautotrophicus), two SAOB (i.e. Tepidanaerobacter acetatoxydans and Thermacetogenium phaeum) and their syntrophic cultivation were assessed under 0.26, 3, 5 and 7 g NH4 (+)-N L(-1). The results showed that some hydrogenotrophic methanogens were equally, or in some cases, more tolerant to high ammonia levels compared to SAOB. Furthermore, a mesophilic hydrogenotrophic methanogen was more sensitive to ammonia toxicity compared to thermophilic methanogens tested in the study, which is contradicting to the general belief that thermophilic methanogens are more vulnerable to high ammonia loads compared to mesophilic. This unexpected finding underlines the fact that the complete knowledge of ammonia inhibition effect on hydrogenotrophic methanogens is still absent. PMID:26490748

  2. Modeling the significance of including C redistribution when determining changes in net carbon storage along a cultivated toposequence

    NASA Astrophysics Data System (ADS)

    Chirinda, Ngonidzashe; Olesen, Jørgen E.; Heckrath, Goswin; Paradelo Pérez, Marcos; Taghizadeh-Toosi, Arezoo

    2016-04-01

    Globally, soil carbon (C) reserves are second only to those in the ocean, and accounts for a significant C reservoir. In the case of arable soils, the quantity of stored C is influenced by various factors (e.g. management practices). Currently, the topography related influences on in-field soil C dynamics remain largely unknown. However, topography is known to influence a multiplicity of factors that regulate C input, storage and redistribution. To understand the patterns and untangle the complexity of soil C dynamics in arable landscapes, our study was conducted with soils from shoulderslope and footslope positions on a 7.1 ha winter wheat field in western Denmark. We first collected soil samples from shoulderslope and footslope positions with various depth intervals down to 100 cm and analyzed them for physical and chemical properties including texture and soil organic C contents. In-situ carbon dioxide (CO2) concentrations were measured at different soil profile depths at both positions for a year. Soil moisture content and temperature at 5 and 40 cm depth was measured continuously. Additionally, surface soil CO2 fluxes at shoulderslope and footslope positions were measured. We then used measurement data collected from the two landscape positions to calibrate the one-dimensional mechanistic model SOILCO2 module of the HYDRUS-1D software package and obtained soil CO2 fluxes from soil profile at two landscape positions. Furthermore, we tested whether the inclusion of vertical and lateral soil C movement improved the modeling of C dynamics in cultivated landscapes. For that, soil profile CO2 fluxes were compared with those obtained using a simple process-based soil whole profile C model, C-TOOL, which was modified to include vertical and lateral movement of C on landscape. Our results highlight the need to consider vertical and lateral soil C movement in the modeling of C dynamics in cultivated landscapes, for better qualification of net carbon storage.

  3. Nickel isotopes and methanogens

    NASA Astrophysics Data System (ADS)

    Neubeck, A.; Ivarsson, M.

    2013-12-01

    Methanogens require Ni for their growth and as a consequence the microbial fractionation of Ni isotopes can be used as a biomarker for activity of methanogenic communities1. Anaerobic laboratory experiments was performed using methanogens to investigate methanogenic growth in a modified nutrient media2 with olivine Fo91 (5g/l) added as an additional mineral nutrient source and as the only H2 provider. One of the investigated methanogens showed an increased growth in the experiments with added olivine. There were also a close relationship between the mobilized Ni and the growth of the methanogen. Ni is an element that previously has been neglected in the study of fossilized microorganisms and their interaction with mineral substrates and, thus, there are no records or published data of Ni in association with microfossils. However, we have detected enrichments of Ni in fossilized microorganisms and ichno-fossils, respectively, from three separate locations. Ni is not present in the host rock in any of the samples. Thus, Ni is present in association with fossilized microorganisms from environments and more extensive analysis is required to understand the magnitude, uptake, preservation and fractionation of Ni in microfossils. In order to analyze Ni isotope fractionation from microbe-mineral interaction, we plan to use a high-resolution Laser-Ablation Time-of-Flight Mass Spectrometer (LMS)3. In situ profile ablation will provide detailed and localized data on fractionation patterns between microfossils and their host rock. Also, this technique will allow us to identify the change in Ni isotopic fractionation in rock samples caused by abiotic and biogenic processes in a faster and easier way and with less risk for contamination compared to the wet chemistry analyses of Ni isotopes. 1. Cameron, V., Vance, D., Archer, C. & House, C. H. A biomarker based on the stable isotopes of nickel. Proceedings of the National Academy of Sciences 106, 10944-10948 (2009). 2. Schn

  4. Method for Indirect Quantification of CH4 Production via H2O Production Using Hydrogenotrophic Methanogens.

    PubMed

    Taubner, Ruth-Sophie; Rittmann, Simon K-M R

    2016-01-01

    Hydrogenotrophic methanogens are an intriguing group of microorganisms from the domain Archaea. Methanogens exhibit extraordinary ecological, biochemical, and physiological characteristics and possess a huge biotechnological potential. Yet, the only possibility to assess the methane (CH4) production potential of hydrogenotrophic methanogens is to apply gas chromatographic quantification of CH4. In order to be able to effectively screen pure cultures of hydrogenotrophic methanogens regarding their CH4 production potential we developed a novel method for indirect quantification of the volumetric CH4 production rate by measuring the volumetric water production rate. This method was established in serum bottles for cultivation of methanogens in closed batch cultivation mode. Water production was estimated by determining the difference in mass increase in a quasi-isobaric setting. This novel CH4 quantification method is an accurate and precise analytical technique, which can be used to rapidly screen pure cultures of methanogens regarding their volumetric CH4 evolution rate. It is a cost effective alternative determining CH4 production of methanogens over CH4 quantification by using gas chromatography, especially if applied as a high throughput quantification method. Eventually, the method can be universally applied for quantification of CH4 production from psychrophilic, thermophilic and hyperthermophilic hydrogenotrophic methanogens. PMID:27199898

  5. Method for Indirect Quantification of CH4 Production via H2O Production Using Hydrogenotrophic Methanogens

    PubMed Central

    Taubner, Ruth-Sophie; Rittmann, Simon K.-M. R.

    2016-01-01

    Hydrogenotrophic methanogens are an intriguing group of microorganisms from the domain Archaea. Methanogens exhibit extraordinary ecological, biochemical, and physiological characteristics and possess a huge biotechnological potential. Yet, the only possibility to assess the methane (CH4) production potential of hydrogenotrophic methanogens is to apply gas chromatographic quantification of CH4. In order to be able to effectively screen pure cultures of hydrogenotrophic methanogens regarding their CH4 production potential we developed a novel method for indirect quantification of the volumetric CH4 production rate by measuring the volumetric water production rate. This method was established in serum bottles for cultivation of methanogens in closed batch cultivation mode. Water production was estimated by determining the difference in mass increase in a quasi-isobaric setting. This novel CH4 quantification method is an accurate and precise analytical technique, which can be used to rapidly screen pure cultures of methanogens regarding their volumetric CH4 evolution rate. It is a cost effective alternative determining CH4 production of methanogens over CH4 quantification by using gas chromatography, especially if applied as a high throughput quantification method. Eventually, the method can be universally applied for quantification of CH4 production from psychrophilic, thermophilic and hyperthermophilic hydrogenotrophic methanogens. PMID:27199898

  6. Greenhouse Gas Emissions and Global Warming Potential of Traditional and Diversified Tropical Rice Rotation Systems including Impacts of Upland Crop Management Practices i.e. Mulching and Inter-crop Cultivation

    NASA Astrophysics Data System (ADS)

    Janz, Baldur; Weller, Sebastian; Kraus, David; Wassmann, Reiner; Butterbach-Bahl, Klaus; Kiese, Ralf

    2016-04-01

    Paddy rice cultivation is increasingly challenged by irrigation water scarcity, while at the same time changes in demand (e.g. changes in diets or increasing demand for biofuels) will feed back on agricultural practices. These factors are changing traditional cropping patterns from flooded double-rice systems to the introduction of well-aerated upland crop systems in the dry season. Emissions of methane (CH4) are expected to decrease, while emissions of nitrous oxide (N2O) will increase and soil organic carbon (SOC) stocks will most likely be volatilized in the form of carbon dioxide (CO2). We measured greenhouse gas (GHG) emissions at the International Rice Research Institute (IRRI) in the Philippines to provide a comparative assessment of the global warming potentials (GWP) as well as yield scaled GWPs of different crop rotations and to evaluate mitigation potentials or risks of new management practices i.e. mulching and inter-crop cultivation. New management practices of mulching and intercrop cultivation will also have the potential to change SOC dynamics, thus can play the key role in contributing to the GWP of upland cropping systems. To present, more than three years of continuous measurement data of CH4 and N2O emissions in double-rice cropping (R-R) and paddy rice rotations diversified with either maize (R-M) or aerobic rice (R-A) in upland cultivation have been collected. Introduction of upland crops in the dry season reduced irrigation water use and CH4 emissions by 66-81% and 95-99%, respectively. Moreover, for practices including upland crops, CH4 emissions in the subsequent wet season with paddy rice were reduced by 54-60%. Although annual N2O emissions increased twice- to threefold in the diversified systems, the strong reduction of CH4 led to a significantly lower (p<0.05) annual GWP (CH4+ N2O) as compared to the traditional double-rice cropping system. Measurements of soil organic carbon contents before and three years after introduction of upland

  7. Dehydropyrrolizidine alkaloids, including monoesters with an unusual esterifying acid, from cultivated Crotalaria juncea (Sunn Hemp cv. 'Tropic Sun')

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Cultivation of Crotalaria juncea L. (Sunn Hemp cv. ‘Tropic Sun’) is recommended as a green manure crop in a rotation cycle to improve soil condition, help control erosion, suppress weeds, and reduce soil nematodes. Because C. juncea belongs to a genus that is known for the production of toxic dehydr...

  8. Light sensitivity of methanogenic archaebacteria

    SciTech Connect

    Olson, K.D.; McMahon, C.W.; Wolfe, R.S. )

    1991-09-01

    Representatives of four families of methanogenic archaebacteria (archaea), Methanobacterium thermoautotrophicum {Delta}H, Methanobacterium thermoautotrophicum Marburg, Methanosarcina acetivorans, Methanococcus voltae, and Methanomicrobium mobile, were found to be light sensitive. The facultative anaerobic eubacteria Escherichia coli and Salmonella typhimurium, however, were tolerant of light when grown anaerobically under identical light conditions. Interference filters were used to show that the growth of the methanogens is inhibited by light in the blue end of the visible spectrum (370 to 430 nm).

  9. Enumeration of methanogens with a focus on fluorescence in situ hybridization

    NASA Astrophysics Data System (ADS)

    Kumar, Sanjay; Dagar, Sumit Singh; Mohanty, Ashok Kumar; Sirohi, Sunil Kumar; Puniya, Monica; Kuhad, Ramesh C.; Sangu, K. P. S.; Griffith, Gareth Wyn; Puniya, Anil Kumar

    2011-06-01

    Methanogens, the members of domain Archaea are potent contributors in global warming. Being confined to the strict anaerobic environment, their direct cultivation as pure culture is quite difficult. Therefore, a range of culture-independent methods have been developed to investigate their numbers, substrate uptake patterns, and identification in complex microbial communities. Unlike other approaches, fluorescence in situ hybridization (FISH) is not only used for faster quantification and accurate identification but also to reveal the physiological properties and spatiotemporal dynamics of methanogens in their natural environment. Aside from the methodological aspects and application of FISH, this review also focuses on culture-dependent and -independent techniques employed in enumerating methanogens along with associated problems. In addition, the combination of FISH with micro-autoradiography that could also be an important tool in investigating the activities of methanogens is also discussed.

  10. Role for acetotrophic methanogens in methanogenic biodegradation of vinyl chloride

    SciTech Connect

    Bradley, P.M.; Chapelle, F.H.

    1999-10-01

    Under methanogenic conditions, stream-bed sediment microorganisms rapidly degraded [1,2-{sup 14}C]vinyl chloride to {sup 14}CH{sub 4} and {sup 14}CO{sub 2}. Amendment with 2-bromoethanesulfonic acid eliminated {sup 14}CH{sub 4} production and decreased {sup 14}CO{sub 2} recovery by an equal molar amount. Results obtained with [{sup 14}C]ethene, [{sup 14}C]acetate, or {sup 14}CO{sub 2} as substrates indicated that acetotrophic methanogens were responsible for the production of {sup 14}CH{sub 4} during biodegradation of [1,2-{sup 14}C]VC.

  11. Role for acetotrophic methanogens in methanogenic biodegradation of vinyl chloride

    USGS Publications Warehouse

    Bradley, P.M.; Chapelle, F.H.

    1999-01-01

    Under methanogenic conditions, stream-bed sediment microorganisms rapidly degraded [1,2-14C]vinyl chloride to 14CH4 and 14CO2. Amendment with 2-bromoethanesulfonic acid eliminated 14CH4 production and decreased 14CO2 recovery by an equal molar amount. Results obtained with [14C]ethene, [14C]acetate, or 14CO2 as substrates indicated that acetotrophic methanogens were responsible for the production of 14CH4 during biodegradation of [1,2-14C]VC.Under methanogenic conditions, stream-bed sediment microorganisms rapidly degraded [1,2-14C]vinyl chloride to 14CH4 and 14CO2. Amendment with 2-bromoethanesulfonic acid eliminated 14CH4 production and decreased 14CO2 recovery by an equal molar amount. Results obtained with [14C]-ethene, [14C]acetate, or 14CO2 as substrates indicated that acetotrophic methanogens were responsible for the production of 14CH4, during biodegradation of [1,2-14C]VC.

  12. Aerobic culture of methanogenic archaea without an external source of hydrogen.

    PubMed

    Khelaifia, S; Lagier, J-C; Nkamga, V D; Guilhot, E; Drancourt, M; Raoult, D

    2016-06-01

    Culturing methanogenic archaea is fastidious, expensive, and requires an external source of hydrogen and carbon dioxide. Until now, these microorganisms have only been cultivated under strictly anaerobic conditions. We previously developed a single versatile culture medium containing sugars and anti-oxydants for cultivating all human known methanogens. Performing aerobic cultures in the presence of Bacteroides thetaiotaomicron, which produces hydrogen, allows for cultivation of Methanobrevibacter smithii which itself produces methane. To obtain colonies, we cultivated M. smithii in an agar plate in the upper part of a double chamber flask with a liquid culture of B. thetaiotaomicron in the lower compartment. We subsequently cultured four other methanogenic species for the first time and successfully isolated 13 strains of M. smithii and nine strains of Methanobrevibacter oralis from 100 stools and 45 oral samples. This procedure allows aerobic isolation and antibiotic susceptibility testing. This changes the ability to routinely study methanogens, which have been neglected in clinical microbiology laboratories and may be useful for biogas production. PMID:27010812

  13. Methanogenic Hydrocarbon Degradation: Evidence from Field and Laboratory Studies.

    PubMed

    Jiménez, Núria; Richnow, Hans H; Vogt, Carsten; Treude, Tina; Krüger, Martin

    2016-01-01

    Microbial transformation of hydrocarbons to methane is an environmentally relevant process taking place in a wide variety of electron acceptor-depleted habitats, from oil reservoirs and coal deposits to contaminated groundwater and deep sediments. Methanogenic hydrocarbon degradation is considered to be a major process in reservoir degradation and one of the main processes responsible for the formation of heavy oil deposits and oil sands. In the absence of external electron acceptors such as oxygen, nitrate, sulfate or Fe(III), fermentation and methanogenesis become the dominant microbial metabolisms. The major end product under these conditions is methane, and the only electron acceptor necessary to sustain the intermediate steps in this process is CO2, which is itself a net product of the overall reaction. We are summarizing the state of the art and recent advances in methanogenic hydrocarbon degradation research. Both the key microbial groups involved as well as metabolic pathways are described, and we discuss the novel insights into methanogenic hydrocarbon-degrading populations studied in laboratory as well as environmental systems enabled by novel cultivation-based and molecular approaches. Their possible implications on energy resources, bioremediation of contaminated sites, deep-biosphere research, and consequences for atmospheric composition and ultimately climate change are also addressed. PMID:26959375

  14. Dynamics of the Methanogenic Archaea in Tropical Estuarine Sediments

    PubMed Central

    Torres-Alvarado, María del Rocío; Fernández, Francisco José; Ramírez Vives, Florina; Varona-Cordero, Francisco

    2013-01-01

    Methanogenesis may represent a key process in the terminal phases of anaerobic organic matter mineralization in sediments of coastal lagoons. The aim of the present work was to study the temporal and spatial dynamics of methanogenic archaea in sediments of tropical coastal lagoons and their relationship with environmental changes in order to determine how these influence methanogenic community. Sediment samples were collected during the dry (February, May, and early June) and rainy seasons (July, October, and November). Microbiological analysis included the quantification of viable methanogenic archaea (MA) with three substrates and the evaluation of kinetic activity from acetate in the presence and absence of sulfate. The environmental variables assessed were temperature, pH, Eh, salinity, sulfate, solids content, organic carbon, and carbohydrates. MA abundance was significantly higher in the rainy season (106–107 cells/g) compared with the dry season (104–106 cells/g), with methanol as an important substrate. At spatial level, MA were detected in the two layers analyzed, and no important variations were observed either in MA abundance or activity. Salinity, sulfate, solids, organic carbon, and Eh were the environmental variables related to methanogenic community. A conceptual model is proposed to explain the dynamics of the MA. PMID:23401664

  15. Adaptation of a methanogenic consortium to arsenite inhibition

    PubMed Central

    Rodriguez-Freire, Lucia; Moore, Sarah E.; Sierra-Alvarez, Reyes; Field, James A.

    2016-01-01

    Arsenic (As) is a ubiquitous metalloid known for its adverse effects to human health. Microorganisms are also impacted by As toxicity, including methanogenic archaea, which can affect the performance of process in which biological activity is required (i.e. stabilization of activated sludge in wastewater treatment plants). The novel ability of a mixed methanogenic granular sludge consortium to adapt to the inhibitory effect of arsenic (As) was investigated by exposing the culture to approximately 0.92 mM of AsIII for 160 d in an arsenate (AsV) reducing bioreactor using ethanol as the electron donor. The results of shaken batch bioassays indicated that the original, unexposed sludge was severely inhibited by arsenite (AsIII) as evidenced by the low 50% inhibition concentrations (IC50) determined, i.e., 19 and 90 μM for acetoclastic- and hydrogenotrophic methanogenesis, respectively. The tolerance of the acetoclastic and hydrogenotrophic methanogens in the sludge to AsIII increased 47-fold (IC50 = 910 μM) and 12-fold (IC50= 1100 μM), respectively, upon long-term exposure to As. In conclusion, the methanogenic community in the granular sludge demonstrated a considerable ability to adapt to the severe inhibitory effects of As after a prolonged exposure period. PMID:26823637

  16. Relating methanogen community structure and anaerobic digester function.

    PubMed

    Bocher, B T W; Cherukuri, K; Maki, J S; Johnson, M; Zitomer, D H

    2015-03-01

    Much remains unknown about the relationships between microbial community structure and anaerobic digester function. However, knowledge of links between community structure and function, such as specific methanogenic activity (SMA) and COD removal rate, are valuable to improve anaerobic bioprocesses. In this work, quantitative structure-activity relationships (QSARs) were developed using multiple linear regression (MLR) to predict SMA using methanogen community structure descriptors for 49 cultures. Community descriptors were DGGE demeaned standardized band intensities for amplicons of a methanogen functional gene (mcrA). First, predictive accuracy of MLR QSARs was assessed using cross validation with training (n = 30) and test sets (n = 19) for glucose and propionate SMA data. MLR equations correlating band intensities and SMA demonstrated good predictability for glucose (q(2) = 0.54) and propionate (q(2) = 0.53). Subsequently, data from all 49 cultures were used to develop QSARs to predict SMA values. Higher intensities of two bands were correlated with higher SMA values; high abundance of methanogens associated with these two bands should be encouraged to attain high SMA values. QSARs are helpful tools to identify key microorganisms or to study and improve many bioprocesses. Development of new, more robust QSARs is encouraged for anaerobic digestion or other bioprocesses, including nitrification, nitritation, denitrification, anaerobic ammonium oxidation, and enhanced biological phosphorus removal. PMID:25562581

  17. STUDIES OF METHANOGENIC BACTERIA IN SLUDGE

    EPA Science Inventory

    Methanogenic bacteria were isolated from mesophilic anaerobic digesters. The isolates were able to utilize H2 and CO2 acetate, formate and methanol, but were not able to metabolize propionate and butyrate. It was shown the propionate and butyrate are not substrates for methanogen...

  18. Enrichment of hydrogenotrophic methanogens in coupling with methane production using an electrochemical bioreactor.

    PubMed

    Jeon, Bo Young; Kim, Sung Yong; Park, Yong Keun; Park, Doo Hyun

    2009-12-01

    Anaerobic digestion sludge was cultivated in an electrochemical bioreactor (ECB) to enrich the hydrogenotrophic methanogens. A modified graphite felt cathode with neutral red (NR-cathode) was charged with electrochemical reducing power generated from a solar cell. The methane and carbon dioxide collected in a Teflon bag from the ECB were more than 80 ml/l of reactant/day and less than 20 ml/l of reactant/day, respectively, whereas the methane and carbon dioxide collected from a conventional bioreactor (CB) was around 40 ml/l of reactant/day, respectively. Moreover, the maximal volume ratios of methane to carbon dioxide (M/C ratio) collected in the Teflon bag from the ECB and CB were 7 and 1, respectively. The most predominant methanogens isolated from the CB on the 20th, 80th, and 150th days of incubation were hydrogenotrophs. The methanogenic diversity analyzed by temperature gradient gel electrophoresis (TGGE) of the 16S rDNA variable region was higher in the ECB than in the CB. The DNA extracted from the TGGE bands was more than 95% homologous with hydrogenotrophic methanogens in the CB. In conclusion, the ECB was demonstrated as a useful system for enriching hydrogenotrophic methanogens and increasing the M/C ratio of the gas product. PMID:20075635

  19. A mechanistic model of hydrogen-methanogen dynamics in the rumen.

    PubMed

    Wang, Yuancheng; Janssen, Peter H; Lynch, Tammy A; Brunt, Bruce van; Pacheco, David

    2016-03-21

    Existing mathematical models to estimate methane production in the rumen are based on calculation of hydrogen balances without considering the presence of methanogens. In this study, a mechanistic model of methane production is proposed that depicts the interaction between hydrogen concentration and methanogens in the rumen. Analytical results show that it meets biological expectations, namely increased fractional passage rate leads to a greater growth rate of methanogens, and a greater steady state hydrogen concentration. This model provides a basis on which to develop a more comprehensive model of methane production in the rumen that includes thermodynamics and feed fermentation pathways. PMID:26780651

  20. Impact of dewatering technologies on specific methanogenic activity.

    PubMed

    Batstone, Damien J; Lu, Yang; Jensen, Paul D

    2015-10-01

    Dewatering methods for recuperative thickening and final dewatering can potentially impact methanogenic activity and microbial community. This influences both the feasibility of recuperative thickening to increase solids residence time within a digester, and the utilisation of dewatered digestate as inoculum for new digesters. Thickening technology can reduce methanogenic activity through either air contact (rotary drum, DAF, or belt filter press), or by lysing cells through shear (centrifuge). To assess this, two plants with recuperative thickening (rotary drum) in their anaerobic digester, and five without recuperative thickening, had specific methanogenic activity tested in all related streams, including dewatering feed, thickened return, final cake, and centrate. All plants had high speed centrifuges for final dewatering. The digester microbial community was also assessed through 16s pyrotag sequencing and subsequent principal component analysis (PCA). The specific methanogenic activity of all samples was in the expected range of 0.2-0.4 gCOD gVS(-1)d(-1). Plants with recuperative thickening did not have lower digester activity. Centrifuge based dewatering had a significant and variable impact on methanogenic activity in all samples, ranging between 20% and 90% decrease but averaging 54%. Rotary drum based recuperative thickening had a far smaller impact on activity, with a 0% per-pass drop in activity in one plant, and a 20% drop in another. However, the presence of recuperative thickening was a major predictor of overall microbial community (PC1, p = 0.0024). Microbial community PC3 (mainly driven by a shift in methanogens) was a strong predictor for sensitivity in activity to shear (p = 0.0005, p = 0.00001 without outlier). The one outlier was related to a plant producing the wettest cake (17% solids). This indicates that high solids is a potential driver of sensitivity to shear, but that a resilient microbial community can also bestow resilience

  1. Diffusional Properties of Methanogenic Granular Sludge: 1H NMR Characterization

    PubMed Central

    Lens, Piet N. L.; Gastesi, Rakel; Vergeldt, Frank; van Aelst, Adriaan C.; Pisabarro, Antonio G.; Van As, Henk

    2003-01-01

    The diffusive properties of anaerobic methanogenic and sulfidogenic aggregates present in wastewater treatment bioreactors were studied using diffusion analysis by relaxation time-separated pulsed-field gradient nuclear magnetic resonance (NMR) spectroscopy and NMR imaging. NMR spectroscopy measurements were performed at 22°C with 10 ml of granular sludge at a magnetic field strength of 0.5 T (20 MHz resonance frequency for protons). Self-diffusion coefficients of H2O in the investigated series of mesophilic aggregates were found to be 51 to 78% lower than the self-diffusion coefficient of free water. Interestingly, self-diffusion coefficients of H2O were independent of the aggregate size for the size fractions investigated. Diffusional transport occurred faster in aggregates growing under nutrient-rich conditions (e.g., the bottom of a reactor) or at high (55°C) temperatures than in aggregates cultivated in nutrient-poor conditions or at low (10°C) temperatures. Exposure of aggregates to 2.5% glutaraldehyde or heat (70 or 90°C for 30 min) modified the diffusional transport up to 20%. In contrast, deactivation of aggregates by HgCl2 did not affect the H2O self-diffusion coefficient in aggregates. Analysis of NMR images of a single aggregate shows that methanogenic aggregates possess a spin-spin relaxation time and self-diffusion coefficient distribution, which are due to both physical (porosity) and chemical (metal sulfide precipitates) factors. PMID:14602624

  2. Acceleration of cellulose degradation and shift of product via methanogenic co-culture of a cellulolytic bacterium with a hydrogenotrophic methanogen.

    PubMed

    Sasaki, Daisuke; Morita, Masahiko; Sasaki, Kengo; Watanabe, Atsushi; Ohmura, Naoya

    2012-10-01

    Although the effects of syntrophic relationships between bacteria and methanogens have been reported in some environments, those on cellulose decomposition using cellulolytic bacteria from methanogenic reactors have not yet been examined. The effects of syntrophic co-culture on the decomposition of a cellulosic material were investigated in a co-culture of Clostridium clariflavum strain CL-1 and the hydrogenotrophic methanogen Methanothermobacter thermautotrophicus strain ΔH and a single-culture of strain CL-1 under thermophilic conditions. In this study, strain CL-1 was newly isolated as a cellulolytic bacterium from a thermophilic methanogenic reactor used for degrading garbage slurry. The degradation efficiency and cell density of strain CL-1 were 2.9- and 2.7-fold higher in the co-culture than in the single-culture after 60 h of incubation, respectively. Acetate, lactate and ethanol were the primary products in both cultures, and the concentration of propionate was low. The content of acetate to total organic acids plus ethanol was 59.3% in the co-culture. However, the ratio decreased to 24.9% in the single-culture, although acetate was the primary product. Therefore, hydrogen scavenging by the hydrogenotrophic methanogen strain ΔH could shift the metabolic pathway to the acetate production pathway in the co-culture. Increases in the cell density and the consequent acceleration of cellulose degradation in the co-culture would be caused by increases in adenosine 5'-triphosphate (ATP) levels, as the acetate production pathway includes ATP generation. Syntrophic cellulose decomposition by the cellulolytic bacteria and hydrogenotrophic methanogens would be the dominant reaction in the thermophilic methanogenic reactor degrading cellulosic materials. PMID:22652087

  3. Environmental controls on methanogen viability in the hydrothermal waters of the El Tatio geyser field, Chile.

    NASA Astrophysics Data System (ADS)

    Franks, M. A.; Bennett, P. C.; Omelon, C.; Engel, A. S.

    2007-12-01

    At the El Tatio geyser field, a unique hydrothermal site located in the Andes Mountains in Chile, methanogenic archaea were found in only two of the hundreds of hydrothermal features. Reported here is an investigation into the environmental and geochemical controls on the distribution of methanogenic archaea. Located in the hyper- arid Atacama Desert, El Tatio waters are characterized by high salinity (95-175mM), Na-Cl type waters and circum-neutral pH (6.5-7), with very low inorganic carbon (0.1-0.5 mM TIC), but very high concentrations of As and Sb (300-700 uM As, 10-30uM Sb). Extensive bacterial mats thrive in most of the shallow run-off streams originating from hydrothermal features. In order to determine geochemical controls on methanogen populations, major and trace elements, including As and Sb speciation and concentrations, were determined using IC and HPLC-ICP-MS methods. The structure of microbial communities was analyzed using MPN enumeration of methanogens, culturing, and phylogenetic analysis using molecular techniques. Here, as in many hydrothermal regions, temperature and geochemical gradients influence the microbial ecology. Results from MPN enumeration indicate methanogen populations are dominated by H2-utilizing (carbonate reducing) archaea at both of the sites, with some acetate-oxidizing archaea present. These sites contain comparatively high DIC concentrations; however, it is unclear whether this is a control or a product of methanogenic archaea. Water quality analyses also show a strong correlation between antimony concentrations and the presence of methanogens; methanogenic archaea being present only at sites with 17 uM Sb concentrations or less.

  4. Methanogenic food web in the gut contents of methane-emitting earthworm Eudrilus eugeniae from Brazil.

    PubMed

    Schulz, Kristin; Hunger, Sindy; Brown, George G; Tsai, Siu M; Cerri, Carlos C; Conrad, Ralf; Drake, Harold L

    2015-08-01

    The anoxic saccharide-rich conditions of the earthworm gut provide an ideal transient habitat for ingested microbes capable of anaerobiosis. It was recently discovered that the earthworm Eudrilus eugeniae from Brazil can emit methane (CH4) and that ingested methanogens might be associated with this emission. The objective of this study was to resolve trophic interactions of bacteria and methanogens in the methanogenic food web in the gut contents of E. eugeniae. RNA-based stable isotope probing of bacterial 16S rRNA as well as mcrA and mrtA (the alpha subunit of methyl-CoM reductase and its isoenzyme, respectively) of methanogens was performed with [(13)C]-glucose as a model saccharide in the gut contents. Concomitant fermentations were augmented by the rapid consumption of glucose, yielding numerous products, including molecular hydrogen (H2), carbon dioxide (CO2), formate, acetate, ethanol, lactate, succinate and propionate. Aeromonadaceae-affiliated facultative aerobes, and obligate anaerobes affiliated to Lachnospiraceae, Veillonellaceae and Ruminococcaceae were associated with the diverse fermentations. Methanogenesis was ongoing during incubations, and (13)C-labeling of CH4 verified that supplemental [(13)C]-glucose derived carbon was dissimilated to CH4. Hydrogenotrophic methanogens affiliated with Methanobacteriaceae and Methanoregulaceae were linked to methanogenesis, and acetogens related to Peptostreptoccocaceae were likewise found to be participants in the methanogenic food web. H2 rather than acetate stimulated methanogenesis in the methanogenic gut content enrichments, and acetogens appeared to dissimilate supplemental H2 to acetate in methanogenic enrichments. These findings provide insight on the processes and associated taxa potentially linked to methanogenesis and the turnover of organic carbon in the alimentary canal of methane-emitting E. eugeniae. PMID:25615437

  5. Community Structure in Methanogenic Enrichments Provides Insight into Syntrophic Interactions in Hydrocarbon-Impacted Environments

    PubMed Central

    Fowler, S. Jane; Toth, Courtney R. A.; Gieg, Lisa M.

    2016-01-01

    The methanogenic biodegradation of crude oil involves the conversion of hydrocarbons to methanogenic substrates by syntrophic bacteria and subsequent methane production by methanogens. Assessing the metabolic roles played by various microbial species in syntrophic communities remains a challenge, but such information has important implications for bioremediation and microbial enhanced energy recovery technologies. Many factors such as changing environmental conditions or substrate variations can influence the composition and biodegradation capabilities of syntrophic microbial communities in hydrocarbon-impacted environments. In this study, a methanogenic crude oil-degrading enrichment culture was successively transferred onto the single long chain fatty acids palmitate or stearate followed by their parent alkanes, hexadecane or octadecane, respectively, in order to assess the impact of different substrates on microbial community composition and retention of hydrocarbon biodegradation genes. 16S rRNA gene sequencing showed that a reduction in substrate diversity resulted in a corresponding loss of microbial diversity, but that hydrocarbon biodegradation genes (such as assA/masD encoding alkylsuccinate synthase) could be retained within a community even in the absence of hydrocarbon substrates. Despite substrate-related diversity changes, all communities were dominated by hydrogenotrophic and acetotrophic methanogens along with bacteria including Clostridium sp., members of the Deltaproteobacteria, and a number of other phyla. Microbial co-occurrence network analysis revealed a dense network of interactions amongst syntrophic bacteria and methanogens that were maintained despite changes in the substrates for methanogenesis. Our results reveal the effect of substrate diversity loss on microbial community diversity, indicate that many syntrophic interactions are stable over time despite changes in substrate pressure, and show that syntrophic interactions amongst

  6. Methanogenic food web in the gut contents of methane-emitting earthworm Eudrilus eugeniae from Brazil

    PubMed Central

    Schulz, Kristin; Hunger, Sindy; Brown, George G; Tsai, Siu M; Cerri, Carlos C; Conrad, Ralf; Drake, Harold L

    2015-01-01

    The anoxic saccharide-rich conditions of the earthworm gut provide an ideal transient habitat for ingested microbes capable of anaerobiosis. It was recently discovered that the earthworm Eudrilus eugeniae from Brazil can emit methane (CH4) and that ingested methanogens might be associated with this emission. The objective of this study was to resolve trophic interactions of bacteria and methanogens in the methanogenic food web in the gut contents of E. eugeniae. RNA-based stable isotope probing of bacterial 16S rRNA as well as mcrA and mrtA (the alpha subunit of methyl-CoM reductase and its isoenzyme, respectively) of methanogens was performed with [13C]-glucose as a model saccharide in the gut contents. Concomitant fermentations were augmented by the rapid consumption of glucose, yielding numerous products, including molecular hydrogen (H2), carbon dioxide (CO2), formate, acetate, ethanol, lactate, succinate and propionate. Aeromonadaceae-affiliated facultative aerobes, and obligate anaerobes affiliated to Lachnospiraceae, Veillonellaceae and Ruminococcaceae were associated with the diverse fermentations. Methanogenesis was ongoing during incubations, and 13C-labeling of CH4 verified that supplemental [13C]-glucose derived carbon was dissimilated to CH4. Hydrogenotrophic methanogens affiliated with Methanobacteriaceae and Methanoregulaceae were linked to methanogenesis, and acetogens related to Peptostreptoccocaceae were likewise found to be participants in the methanogenic food web. H2 rather than acetate stimulated methanogenesis in the methanogenic gut content enrichments, and acetogens appeared to dissimilate supplemental H2 to acetate in methanogenic enrichments. These findings provide insight on the processes and associated taxa potentially linked to methanogenesis and the turnover of organic carbon in the alimentary canal of methane-emitting E. eugeniae. PMID:25615437

  7. Community Structure in Methanogenic Enrichments Provides Insight into Syntrophic Interactions in Hydrocarbon-Impacted Environments.

    PubMed

    Fowler, S Jane; Toth, Courtney R A; Gieg, Lisa M

    2016-01-01

    The methanogenic biodegradation of crude oil involves the conversion of hydrocarbons to methanogenic substrates by syntrophic bacteria and subsequent methane production by methanogens. Assessing the metabolic roles played by various microbial species in syntrophic communities remains a challenge, but such information has important implications for bioremediation and microbial enhanced energy recovery technologies. Many factors such as changing environmental conditions or substrate variations can influence the composition and biodegradation capabilities of syntrophic microbial communities in hydrocarbon-impacted environments. In this study, a methanogenic crude oil-degrading enrichment culture was successively transferred onto the single long chain fatty acids palmitate or stearate followed by their parent alkanes, hexadecane or octadecane, respectively, in order to assess the impact of different substrates on microbial community composition and retention of hydrocarbon biodegradation genes. 16S rRNA gene sequencing showed that a reduction in substrate diversity resulted in a corresponding loss of microbial diversity, but that hydrocarbon biodegradation genes (such as assA/masD encoding alkylsuccinate synthase) could be retained within a community even in the absence of hydrocarbon substrates. Despite substrate-related diversity changes, all communities were dominated by hydrogenotrophic and acetotrophic methanogens along with bacteria including Clostridium sp., members of the Deltaproteobacteria, and a number of other phyla. Microbial co-occurrence network analysis revealed a dense network of interactions amongst syntrophic bacteria and methanogens that were maintained despite changes in the substrates for methanogenesis. Our results reveal the effect of substrate diversity loss on microbial community diversity, indicate that many syntrophic interactions are stable over time despite changes in substrate pressure, and show that syntrophic interactions amongst

  8. Sensitivity and adaptability of methanogens to perchlorates: Implications for life on Mars

    NASA Astrophysics Data System (ADS)

    Kral, Timothy A.; Goodhart, Timothy H.; Harpool, Joshua D.; Hearnsberger, Christopher E.; McCracken, Graham L.; McSpadden, Stanley W.

    2016-01-01

    In 2008, the Mars Phoenix Lander discovered perchlorate at its landing site, and in 2012, the Curiosity rover confirmed the presence of perchlorate on Mars. The research reported here was designed to determine if certain methanogens could grow in the presence of three different perchlorate salt solutions. The methanogens tested were Methanothermobacter wolfeii, Methanosarcina barkeri, Methanobacterium formicicum and Methanococcus maripaludis. Media were prepared containing 0%, 0.5%, 1.0%, 2%, 5% and 10% wt/vol magnesium perchlorate, sodium perchlorate, or calcium perchlorate. Organisms were inoculated into their respective media followed by incubation at each organism's growth temperature. Methane production, commonly used to measure methanogen growth, was measured by gas chromatography of headspace gas samples. Methane concentrations varied with species and perchlorate salt tested. However, all four methanogens produced substantial levels of methane in the presence of up to 1.0% perchlorate, but not higher. The standard procedure for growing methanogens typically includes sodium sulfide, a reducing agent, to reduce residual molecular oxygen. However, the sodium sulfide may have been reducing the perchlorate, thus allowing for growth of the methanogens. To investigate this possibility, experiments were conducted where stainless steel nails were used instead of sodium sulfide as the reducing agent. Prior to the addition of perchlorate and inoculation, the nails were removed from the liquid medium. Just as in the prior experiments, the methanogens produced methane at comparable levels to those seen with sodium sulfide as the reductant, indicating that sodium sulfide did not reduce the perchlorate to any significant extent. Additionally, cells metabolizing in 1% perchlorate were transferred to 2%, cells metabolizing in 2% were transferred to 5%, and finally cells metabolizing in 5% were transferred to 10%. All four species produced methane at 2% and 5%, but not 10

  9. Methanogenic archaea are globally ubiquitous in aerated soils and become active under wet anoxic conditions

    PubMed Central

    Angel, Roey; Claus, Peter; Conrad, Ralf

    2012-01-01

    The prototypical representatives of the Euryarchaeota—the methanogens—are oxygen sensitive and are thought to occur only in highly reduced, anoxic environments. However, we found methanogens of the genera Methanosarcina and Methanocella to be present in many types of upland soils (including dryland soils) sampled globally. These methanogens could be readily activated by incubating the soils as slurry under anoxic conditions, as seen by rapid methane production within a few weeks, without any additional carbon source. Analysis of the archaeal 16S ribosomal RNA gene community profile in the incubated samples through terminal restriction fragment length polymorphism and quantification through quantitative PCR indicated dominance of Methanosarcina, whose gene copy numbers also correlated with methane production rates. Analysis of the δ13C of the methane further supported this, as the dominant methanogenic pathway was in most cases aceticlastic, which Methanocella cannot perform. Sequences of the key methanogenic enzyme methyl coenzyme M reductase retrieved from the soil samples before incubation confirmed that Methanosarcina and Methanocella are the dominant methanogens, though some sequences of Methanobrevibacter and Methanobacterium were also detected. The global occurrence of only two active methanogenic archaea supports the hypothesis that these are autochthonous members of the upland soil biome and are well adapted to their environment. PMID:22071343

  10. Stereochemical studies of acyclic isoprenoids-XII. Lipids of methanogenic bacteria and possible contributions to sediments

    USGS Publications Warehouse

    Risatti, J.B.; Rowland, S.J.; Yon, D.A.; Maxwell, J.R.

    1984-01-01

    Abundant volatile lipids of Methanobacterium thermoautotrophicum and Methanosarcina barkeri include isoprenoid hydrocarbons (??? C30), and C15, C20 and C25 isoprenoid alcohols. M. barkeri contains 2,6,10,15,19-pentamethyleicosane, whose relative stereochemistry is the same as found in marine sediments, indicating that it is a marker of methanogenic activity. The C20, C30 and C25 alkenes in M. thermoautotrophicum also have a preferred sterochemistry; the latter have the 2,6,10,14,18-pentamethyleicosanyl skeleton, suggesting that the alkane in marine sediments may derive from methanogens. The stereochemistry of squalane in a marine sediment is also compatible with an origin in methanogens; in contrast, the stereochemistry of pristane in M. thermoautotrophicum indicates a fossil fuel contaminant origin, suggesting that this and certain other alkanes reported in archaebacteria might also be of contaminant origin. There is, therefore, little evidence at present that the pristane in immature marine sediments originates in methanogens. The C15 and C20 saturated alcohols in M. thermoautotrophicum have mainly the all-R configuration. If this is generally true for methanogens, the C20 alcohol in the Messel shale may originate mainly from methanogens, whereas that in the Green River shale may originate mainly from photosynthetic organisms. ?? 1984.

  11. Molecular analysis of methanogens involved in methanogenic degradation of tetramethylammonium hydroxide in full-scale bioreactors.

    PubMed

    Whang, Liang-Ming; Hu, Tai-Ho; Liu, Pao-Wen Grace; Hung, Yu-Ching; Fukushima, Toshikazu; Wu, Yi-Ju; Chang, Shao-Hsiung

    2015-02-01

    This study investigated methanogenic communities involved in degradation of tetramethylammonium hydroxide (TMAH) in three full-scale bioreactors treating TMAH-containing wastewater. Based on the results of terminal-restriction fragment-length polymorphism (T-RFLP) and quantitative PCR analyses targeting the methyl-coenzyme M reductase alpha subunit (mcrA) genes retrieved from three bioreactors, Methanomethylovorans and Methanosarcina were the dominant methanogens involved in the methanogenic degradation of TMAH in the bioreactors. Furthermore, batch experiments were conducted to evaluate mcrA messenger RNA (mRNA) expression during methanogenic TMAH degradation, and the results indicated that a higher level of TMAH favored mcrA mRNA expression by Methansarcina, while Methanomethylovorans could only express considerable amount of mcrA mRNA at a lower level of TMAH. These results suggest that Methansarcina is responsible for methanogenic TMAH degradation at higher TMAH concentrations, while Methanomethylovorans may be important at a lower TMAH condition. PMID:25261128

  12. Pulp mill wastewater sediment reveals novel methanogenic and cellulolytic populations.

    PubMed

    Yang, Chunyu; Wang, Wei; Du, Miaofen; Li, Chunfang; Ma, Cuiqing; Xu, Ping

    2013-02-01

    Pulp mill wastewater generated from wheat straw is characterized as high alkalinity and very high COD pollution load. A naturally developed microbial community in a pulp mill wastewater storage pool that had been disused were investigated in this study. Owing to natural evaporation and a huge amount of lignocellulose's deposition, the wastewater sediment contains high concentrations of organic matters and sodium ions, but low concentrations of chloride and carbonate. The microbiota inhabiting especially anaerobic community, including methanogenic arhcaea and cellulolytic species, was studied. All archaeal sequences fall into 2 clusters of family Halobacteriaceae and methanogenic archaeon in the phylum Euryarchaeota. In the methanogenic community, phylogenetic analysis of methyl coenzyme M reductase A (mcrA) genes targeted to novel species in genus Methanoculleus or novel genus of order Methanomicrobiales. The predominance of Methanomicrobiales suggests that methanogenesis in this system might be driven by the hydrogenotrophic pathway. As the important primary fermenter for methane production, the cellulolytic community of enzyme GHF48 was found to be dominated by narrower breadth of novel clostridial cellulase genes. Novel anoxic functional members in such extreme sediment provide the possibility of enhancing the efficiency of anoxic treatment of saline and alkaline wastewaters, as well as benefiting to the biomass transformation and biofuel production processes. PMID:23228889

  13. Methanogenic paraffin degradation proceeds via alkane addition to fumarate by 'Smithella' spp. mediated by a syntrophic coupling with hydrogenotrophic methanogens.

    PubMed

    Wawrik, Boris; Marks, Christopher R; Davidova, Irene A; McInerney, Michael J; Pruitt, Shane; Duncan, Kathleen E; Suflita, Joseph M; Callaghan, Amy V

    2016-09-01

    Anaerobic microbial biodegradation of recalcitrant, water-insoluble substrates, such as paraffins, presents unique metabolic challenges. To elucidate this process, a methanogenic consortium capable of mineralizing long-chain n-paraffins (C28 -C50 ) was enriched from San Diego Bay sediment. Analysis of 16S rRNA genes indicated the dominance of Syntrophobacterales (43%) and Methanomicrobiales (26%). Metagenomic sequencing allowed draft genome assembly of dominant uncultivated community members belonging to the bacterial genus Smithella and the archaeal genera Methanoculleus and Methanosaeta. Five contigs encoding homologs of the catalytic subunit of alkylsuccinate synthase (assA) were detected. Additionally, mRNA transcripts for these genes, including a homolog binned within the 'Smithella' sp. SDB genome scaffold, were detected via RT-PCR, implying that paraffins are activated via 'fumarate addition'. Metabolic reconstruction and comparison with genome scaffolds of uncultivated n-alkane degrading 'Smithella' spp. are consistent with the hypothesis that syntrophically growing 'Smithella' spp. may achieve reverse electron transfer by coupling the reoxidation of ETFred to a membrane-bound FeS oxidoreductase functioning as an ETF:menaquinone oxidoreductase. Subsequent electron transfer could proceed via a periplasmic formate dehydrogenase and/or hydrogenase, allowing energetic coupling to hydrogenotrophic methanogens such as Methanoculleus. Ultimately, these data provide fundamental insight into the energy conservation mechanisms that dictate interspecies interactions salient to methanogenic alkane mineralization. PMID:27198766

  14. Novel energetic coupling in the methanogenic bacteria

    SciTech Connect

    Carper, S.W.; Rogers, K.R.; Lancaster, J.R. Jr.

    1986-05-01

    The Archaebacteria are a diverse group of organisms which have been proposed to be a third major line of descent, separate and distinct from the prokaryotes and eukaryotes. Among the unique bioenergetic systems in this group are the light-driven ion pumps bacteriorhodopsin and halorhodopsin, present in the halobacteria. The methanogens are a major member of the Archaebacteria, and couple ATP synthesis to electron transfer to generate methane from several simple substrates. The authors report here studies on the basic overall scheme of energetics in these unique organisms, including the following observations: ATP synthesis coupled to electron transfer does not require the obligatory intermediacy of a transmembrane ion gradient, but appears to occur by a direct mechanism. Intracellular ion homeostasis is maintained by a sodium cycle, involving an electrogenic sodium-translocating ATPase. Based on EPR examinations of crude preparations in the presence of physiological electron donors or acceptors, a minimal electron transfer chain is proposed, involving nickel, flavin, and at least three iron-sulfur centers.

  15. Considerations in the use of fluorescence in situ hybridization (FISH) and confocal laser scanning microscopy to characterize rumen methanogens and define their spatial distributions.

    PubMed

    Valle, Edith R; Henderson, Gemma; Janssen, Peter H; Cox, Faith; Alexander, Trevor W; McAllister, Tim A

    2015-06-01

    In this study, methanogen-specific coenzyme F420 autofluorescence and confocal laser scanning microscopy were used to identify rumen methanogens and define their spatial distribution in free-living, biofilm-, and protozoa-associated microenvironments. Fluorescence in situ hybridization (FISH) with temperature-controlled hybridization was used in an attempt to describe methanogen diversity. A heat pretreatment (65 °C, 1 h) was found to be a noninvasive method to increase probe access to methanogen RNA targets. Despite efforts to optimize FISH, 16S rRNA methanogen-specific probes, including Arch915, bound to some cells that lacked F420, possibly identifying uncharacterized Methanomassiliicoccales or reflecting nonspecific binding to other members of the rumen bacterial community. A probe targeting RNA from the methanogenesis-specific methyl coenzyme M reductase (mcr) gene was shown to detect cultured Methanosarcina cells with signal intensities comparable to those of 16S rRNA probes. However, the probe failed to hybridize with the majority of F420-emitting rumen methanogens, possibly because of differences in cell wall permeability among methanogen species. Methanogens were shown to integrate into microbial biofilms and to exist as ecto- and endosymbionts with rumen protozoa. Characterizing rumen methanogens and defining their spatial distribution may provide insight into mitigation strategies for ruminal methanogenesis. PMID:25924182

  16. Co-occurence of Crenarchaeota, Thermoplasmata and methanogens in anaerobic sludge digesters.

    PubMed

    Chouari, Rakia; Guermazi, Sonda; Sghir, Abdelghani

    2015-05-01

    16S rRNA Crenarchaeota and Thermoplasmata sequences retrieved from 22 anaerobic digesters were analysed. 4.8 and 0.53 % of archaeal sequences were simultaneously affiliated to these lineages. A core of 2 operational taxonomic units (OTUs) representing 0.6 to -33.6 % of all archaeal sequences were defined for the Crenarchaeotes and identified to already known but not yet cultivable organisms in almost half of the digesters sampled. For the Thermoplasmata, apparently less abundant with 0.7 to -4.7 % of the archaeal sequences, 3 OTUs were identified. We showed here that Crenarchaeotes coexist with methanogens and are particularly abundant when Arch I lineage (also called WSA2 by Hugenholtz) is dominant in digesters. Moreover, Thermoplasmata were detected when Crenarchaeota were present. Interactions between methanogens, Crenarchaeotea and Thermoplamata were thus discussed. PMID:25739565

  17. Ether polar lipids of methanogenic bacteria: structures, comparative aspects, and biosyntheses.

    PubMed Central

    Koga, Y; Nishihara, M; Morii, H; Akagawa-Matsushita, M

    1993-01-01

    Complete structures of nearly 40 ether polar lipids from seven species of methanogens have been elucidated during the past 10 years. Three kinds of variations of core lipids, macrocyclic archaeol and two hydroxyarchaeols, were identified, in addition to the usual archaeol and caldarchaeol (for the nomenclature of archaeal [archaebacterial] ether lipids, see the text). Polar head groups of methanogen phospholipids include ethanolamine, serine, inositol, N-acetylglucosamine, dimethyl- and trimethylaminopentanetetrol, and glucosaminylinositol. Glucose is the sole hexose moiety of glycolipids in most methanogens, and galactose and mannose have been found in a few species. Methanogen lipids are characterized by their diversity in phosphate-containing polar head groups and core lipids, which in turn can be used for chemotaxonomy of methanogens. This was shown by preliminary simplified analyses of lipid component residues. Core lipid analysis by high-pressure liquid chromatography provides a method of determining the methanogenic biomass in natural samples. There has been significant progress in the biosynthetic studies of methanogen lipids in recent years. In vivo incorporation experiments have led to delineation of the outline of the synthetic route of the diphytanylglycerol ether core. The mechanisms of biosynthesis of tetraether lipids and various polar lipids, and cell-free systems of either lipid synthesis, however, remain to be elucidated. The significance and the origin of archaeal ether lipids is discussed in terms of the lipid composition of bacteria living in a wide variety of environments, the oxygen requirement for biosynthesis of hydrocarbon chains, and the physicochemical properties and functions of lipids as membrane constituents. PMID:8464404

  18. Ether polar lipids of methanogenic bacteria: structures, comparative aspects, and biosyntheses.

    PubMed

    Koga, Y; Nishihara, M; Morii, H; Akagawa-Matsushita, M

    1993-03-01

    Complete structures of nearly 40 ether polar lipids from seven species of methanogens have been elucidated during the past 10 years. Three kinds of variations of core lipids, macrocyclic archaeol and two hydroxyarchaeols, were identified, in addition to the usual archaeol and caldarchaeol (for the nomenclature of archaeal [archaebacterial] ether lipids, see the text). Polar head groups of methanogen phospholipids include ethanolamine, serine, inositol, N-acetylglucosamine, dimethyl- and trimethylaminopentanetetrol, and glucosaminylinositol. Glucose is the sole hexose moiety of glycolipids in most methanogens, and galactose and mannose have been found in a few species. Methanogen lipids are characterized by their diversity in phosphate-containing polar head groups and core lipids, which in turn can be used for chemotaxonomy of methanogens. This was shown by preliminary simplified analyses of lipid component residues. Core lipid analysis by high-pressure liquid chromatography provides a method of determining the methanogenic biomass in natural samples. There has been significant progress in the biosynthetic studies of methanogen lipids in recent years. In vivo incorporation experiments have led to delineation of the outline of the synthetic route of the diphytanylglycerol ether core. The mechanisms of biosynthesis of tetraether lipids and various polar lipids, and cell-free systems of either lipid synthesis, however, remain to be elucidated. The significance and the origin of archaeal ether lipids is discussed in terms of the lipid composition of bacteria living in a wide variety of environments, the oxygen requirement for biosynthesis of hydrocarbon chains, and the physicochemical properties and functions of lipids as membrane constituents. PMID:8464404

  19. Identification and quantification of methanogenic archaea in adult chicken ceca

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Methanogens, members of the domain Archaea, have been isolated from various animals but few reports exists regarding the isolation of methanogens from chicken, goose, and turkey feces. By using molecular methods for the identification and quantification of methanogenic archea in adult chicken ceca,...

  20. Cattle Manure Enhances Methanogens Diversity and Methane Emissions Compared to Swine Manure under Rice Paddy.

    PubMed

    Kim, Sang Yoon; Pramanik, Prabhat; Bodelier, Paul L E; Kim, Pil Joo

    2014-01-01

    Livestock manures are broadly used in agriculture to improve soil quality. However, manure application can increase the availability of organic carbon, thereby facilitating methane (CH4) production. Cattle and swine manures are expected to have different CH4 emission characteristics in rice paddy soil due to the inherent differences in composition as a result of contrasting diets and digestive physiology between the two livestock types. To compare the effect of ruminant and non-ruminant animal manure applications on CH4 emissions and methanogenic archaeal diversity during rice cultivation (June to September, 2009), fresh cattle and swine manures were applied into experimental pots at 0, 20 and 40 Mg fresh weight (FW) ha-1 in a greenhouse. Applications of manures significantly enhanced total CH4 emissions as compared to chemical fertilization, with cattle manure leading to higher emissions than swine manure. Total organic C contents in cattle (466 g kg-1) and swine (460 g kg-1) manures were of comparable results. Soil organic C (SOC) contents were also similar between the two manure treatments, but dissolved organic C (DOC) was significantly higher in cattle than swine manure. The mcrA gene copy numbers were significantly higher in cattle than swine manure. Diverse groups of methanogens which belong to Methanomicrobiaceae were detected only in cattle-manured but not in swine-manured soil. Methanogens were transferred from cattle manure to rice paddy soils through fresh excrement. In conclusion, cattle manure application can significantly increase CH4 emissions in rice paddy soil during cultivation, and its pretreatment to suppress methanogenic activity without decreasing rice productivity should be considered. PMID:25494364

  1. Cattle Manure Enhances Methanogens Diversity and Methane Emissions Compared to Swine Manure under Rice Paddy

    PubMed Central

    Kim, Sang Yoon; Pramanik, Prabhat; Bodelier, Paul L. E.; Kim, Pil Joo

    2014-01-01

    Livestock manures are broadly used in agriculture to improve soil quality. However, manure application can increase the availability of organic carbon, thereby facilitating methane (CH4) production. Cattle and swine manures are expected to have different CH4 emission characteristics in rice paddy soil due to the inherent differences in composition as a result of contrasting diets and digestive physiology between the two livestock types. To compare the effect of ruminant and non-ruminant animal manure applications on CH4 emissions and methanogenic archaeal diversity during rice cultivation (June to September, 2009), fresh cattle and swine manures were applied into experimental pots at 0, 20 and 40 Mg fresh weight (FW) ha−1 in a greenhouse. Applications of manures significantly enhanced total CH4 emissions as compared to chemical fertilization, with cattle manure leading to higher emissions than swine manure. Total organic C contents in cattle (466 g kg−1) and swine (460 g kg−1) manures were of comparable results. Soil organic C (SOC) contents were also similar between the two manure treatments, but dissolved organic C (DOC) was significantly higher in cattle than swine manure. The mcrA gene copy numbers were significantly higher in cattle than swine manure. Diverse groups of methanogens which belong to Methanomicrobiaceae were detected only in cattle-manured but not in swine-manured soil. Methanogens were transferred from cattle manure to rice paddy soils through fresh excrement. In conclusion, cattle manure application can significantly increase CH4 emissions in rice paddy soil during cultivation, and its pretreatment to suppress methanogenic activity without decreasing rice productivity should be considered. PMID:25494364

  2. Methanogens at the top of the world: occurrence and potential activity of methanogens in newly deglaciated soils in high-altitude cold deserts in the Western Himalayas

    PubMed Central

    Aschenbach, Katrin; Conrad, Ralf; Řeháková, Klára; Doležal, Jiří; Janatková, Kateřina; Angel, Roey

    2013-01-01

    Methanogens typically occur in reduced anoxic environments. However, in recent studies it has been shown that many aerated upland soils, including desert soils also host active methanogens. Here we show that soil samples from high-altitude cold deserts in the western Himalayas (Ladakh, India) produce CH4 after incubation as slurry under anoxic conditions at rates comparable to those of hot desert soils. Samples of matured soil from three different vegetation belts (arid, steppe, and subnival) were compared with younger soils originating from frontal and lateral moraines of receding glaciers. While methanogenic rates were higher in the samples from matured soils, CH4 was also produced in the samples from the recently deglaciated moraines. In both young and matured soils, those covered by a biological soil crust (biocrust) were more active than their bare counterparts. Isotopic analysis showed that in both cases CH4 was initially produced from H2/CO2 but later mostly from acetate. Analysis of the archaeal community in the in situ soil samples revealed a clear dominance of sequences related to Thaumarchaeota, while the methanogenic community comprised only a minor fraction of the archaeal community. Similar to other aerated soils, the methanogenic community was comprised almost solely of the genera Methanosarcina and Methanocella, and possibly also Methanobacterium in some cases. Nevertheless, ~103 gdw−1 soil methanogens were already present in the young moraine soil together with cyanobacteria. Our results demonstrate that Methanosarcina and Methanocella not only tolerate atmospheric oxygen but are also able to survive in these harsh cold environments. Their occurrence in newly deglaciated soils shows that they are early colonizers of desert soils, similar to cyanobacteria, and may play a role in the development of desert biocrusts. PMID:24348469

  3. Methanogens at the top of the world: occurrence and potential activity of methanogens in newly deglaciated soils in high-altitude cold deserts in the Western Himalayas.

    PubMed

    Aschenbach, Katrin; Conrad, Ralf; Reháková, Klára; Doležal, Jiří; Janatková, Kateřina; Angel, Roey

    2013-01-01

    Methanogens typically occur in reduced anoxic environments. However, in recent studies it has been shown that many aerated upland soils, including desert soils also host active methanogens. Here we show that soil samples from high-altitude cold deserts in the western Himalayas (Ladakh, India) produce CH4 after incubation as slurry under anoxic conditions at rates comparable to those of hot desert soils. Samples of matured soil from three different vegetation belts (arid, steppe, and subnival) were compared with younger soils originating from frontal and lateral moraines of receding glaciers. While methanogenic rates were higher in the samples from matured soils, CH4 was also produced in the samples from the recently deglaciated moraines. In both young and matured soils, those covered by a biological soil crust (biocrust) were more active than their bare counterparts. Isotopic analysis showed that in both cases CH4 was initially produced from H2/CO2 but later mostly from acetate. Analysis of the archaeal community in the in situ soil samples revealed a clear dominance of sequences related to Thaumarchaeota, while the methanogenic community comprised only a minor fraction of the archaeal community. Similar to other aerated soils, the methanogenic community was comprised almost solely of the genera Methanosarcina and Methanocella, and possibly also Methanobacterium in some cases. Nevertheless, ~10(3) gdw(-1) soil methanogens were already present in the young moraine soil together with cyanobacteria. Our results demonstrate that Methanosarcina and Methanocella not only tolerate atmospheric oxygen but are also able to survive in these harsh cold environments. Their occurrence in newly deglaciated soils shows that they are early colonizers of desert soils, similar to cyanobacteria, and may play a role in the development of desert biocrusts. PMID:24348469

  4. Quantitative detection of syntrophic fatty acid-degrading bacterial communities in methanogenic environments.

    PubMed

    Mathai, Prince P; Zitomer, Daniel H; Maki, James S

    2015-06-01

    In methanogenic habitats, volatile fatty acids (VFA), such as propionate and butyrate, are major intermediates in organic matter degradation. VFA are further metabolized to H(2), acetate and CO(2) by syntrophic fatty acid-degrading bacteria (SFAB) in association with methanogenic archaea. Despite their indispensable role in VFA degradation, little is known about SFAB abundance and their environmental distribution. To facilitate ecological studies, we developed four novel genus-specific quantitative PCR (qPCR) assays, with primer sets targeting known SFAB: Syntrophobacter, Smithella, Pelotomaculum and Syntrophomonas. Primer set specificity was confirmed using in silico and experimental (target controls, clone libraries and melt-curve analysis) approaches. These qPCR assays were applied to quantify SFAB in a variety of mesophilic methanogenic habitats, including a laboratory propionate enrichment culture, pilot- and full-scale anaerobic reactors, cow rumen, horse faeces, an experimental rice paddy soil, a bog stream and swamp sediments. The highest SFAB 16S rRNA gene copy numbers were found in the propionate enrichment culture and anaerobic reactors, followed by the bog stream and swamp sediment samples. In addition, it was observed that SFAB and methanogen abundance varied with reactor configuration and substrate identity. To our knowledge, this research represents the first comprehensive study to quantify SFAB in methanogenic habitats using qPCR-based methods. These molecular tools will help investigators better understand syntrophic microbial communities in engineered and natural environments. PMID:25814038

  5. Growth of Methanogenic Bacteria in Pure Culture with 2-Propanol and Other Alcohols as Hydrogen Donors

    PubMed Central

    Widdel, Friedrich

    1986-01-01

    Two types of mesophilic, methanogenic bacteria were isolated in pure culture from anaerobic freshwater and marine mud with 2-propanol as the hydrogen donor. The freshwater strain (SK) was a Methanospirillum species, the marine, salt-requiring strain (CV), which had irregular coccoid cells, resembled Methanogenium sp. Stoichiometric measurements revealed formation of 1 mol of CH4 by CO2 reduction, with 4 mol of 2-propanol being converted to acetone. In addition to 2-propanol, the isolates used 2-butanol, H2, or formate but not methanol or polyols. Acetate did not serve as an energy substrate but was necessary as a carbon source. Strain CV also oxidized ethanol or 1-propanol to acetate or propionate, respectively; growth on the latter alcohols was slower, but final cell densities were about threefold higher than on 2-propanol. Both strains grew well in defined, bicarbonate-buffered, sulfide-reduced media. For cultivation of strain CV, additions of biotin, vitamin B12, and tungstate were necessary. The newly isolated strains are the first methanogens that were shown to grow in pure culture with alcohols other than methanol. Bioenergetic aspects of secondary and primary alcohol utilization by methanogens are discussed. Images PMID:16347050

  6. Response of Methanogens in Arctic Sediments to Temperature and Methanogenic Substrate Availability

    PubMed Central

    Blake, Lynsay I.; Tveit, Alexander; Øvreås, Lise; Head, Ian M.; Gray, Neil D.

    2015-01-01

    Although cold environments are major contributors to global biogeochemical cycles, comparatively little is known about their microbial community function, structure, and limits of activity. In this study a microcosm based approach was used to investigate the effects of temperature, and methanogenic substrate amendment, (acetate, methanol and H2/CO2) on methanogen activity and methanogen community structure in high Arctic wetlands (Solvatnet and Stuphallet, Svalbard). Methane production was not detected in Stuphallet sediment microcosms (over a 150 day period) and occurred within Solvatnet sediments microcosms (within 24 hours) at temperatures from 5 to 40°C, the maximum temperature being at far higher than in situ maximum temperatures (which range from air temperatures of -1.4 to 14.1°C during summer months). Distinct responses were observed in the Solvatnet methanogen community under different short term incubation conditions. Specifically, different communities were selected at higher and lower temperatures. At lower temperatures (5°C) addition of exogenous substrates (acetate, methanol or H2/CO2) had no stimulatory effect on the rate of methanogenesis or on methanogen community structure. The community in these incubations was dominated by members of the Methanoregulaceae/WCHA2-08 family-level group, which were most similar to the psychrotolerant hydrogenotrophic methanogen Methanosphaerula palustris strain E1-9c. In contrast, at higher temperatures, substrate amendment enhanced methane production in H2/CO2 amended microcosms, and played a clear role in structuring methanogen communities. Specifically, at 30°C members of the Methanoregulaceae/WCHA2-08 predominated following incubation with H2/CO2, and Methanosarcinaceaeand Methanosaetaceae were enriched in response to acetate addition. These results may indicate that in transiently cold environments, methanogen communities can rapidly respond to moderate short term increases in temperature, but not

  7. Volatile hydrocarbons inhibit methanogenic crude oil degradation

    PubMed Central

    Sherry, Angela; Grant, Russell J.; Aitken, Carolyn M.; Jones, D. Martin; Head, Ian M.; Gray, Neil D.

    2014-01-01

    Methanogenic degradation of crude oil in subsurface sediments occurs slowly, but without the need for exogenous electron acceptors, is sustained for long periods and has enormous economic and environmental consequences. Here we show that volatile hydrocarbons are inhibitory to methanogenic oil biodegradation by comparing degradation of an artificially weathered crude oil with volatile hydrocarbons removed, with the same oil that was not weathered. Volatile hydrocarbons (nC5–nC10, methylcyclohexane, benzene, toluene, and xylenes) were quantified in the headspace of microcosms. Aliphatic (n-alkanes nC12–nC34) and aromatic hydrocarbons (4-methylbiphenyl, 3-methylbiphenyl, 2-methylnaphthalene, 1-methylnaphthalene) were quantified in the total hydrocarbon fraction extracted from the microcosms. 16S rRNA genes from key microorganisms known to play an important role in methanogenic alkane degradation (Smithella and Methanomicrobiales) were quantified by quantitative PCR. Methane production from degradation of weathered oil in microcosms was rapid (1.1 ± 0.1 μmol CH4/g sediment/day) with stoichiometric yields consistent with degradation of heavier n-alkanes (nC12–nC34). For non-weathered oil, degradation rates in microcosms were significantly lower (0.4 ± 0.3 μmol CH4/g sediment/day). This indicated that volatile hydrocarbons present in the non-weathered oil inhibit, but do not completely halt, methanogenic alkane biodegradation. These findings are significant with respect to rates of biodegradation of crude oils with abundant volatile hydrocarbons in anoxic, sulphate-depleted subsurface environments, such as contaminated marine sediments which have been entrained below the sulfate-reduction zone, as well as crude oil biodegradation in petroleum reservoirs and contaminated aquifers. PMID:24765087

  8. Microbiology and biochemistry of the methanogenic archaeobacteria

    NASA Astrophysics Data System (ADS)

    Abbanat, Darren R.; Aceti, David J.; Baron, Stephen F.; Terlesky, Katherine C.; Ferry, James C.

    The methane producing bacteria area diverse group of organisms that function in nature with other groups of strictly anaerobic bacteria to convert complex organic matter to methane and carbon dioxide. The methanogens belong to the archaeobacteria, a third primary kingdom distinct from all other procaryotes (eubacteria) and eucaryotes. The distinction is based on the unique structures of cell wall and membrane components present in archaeobacteria, as well as differences in the highly conserved 16s rRNA sequences among the three kingdoms. In addition, the methanogens contain several novel cofactors that function as one-carbon carriers during the reduction of carbon dioxide to methane with electrons derived from the oxidation of H2 or formate. Methanogens also convert acetate to methane by a pathway distinct from that for carbon dioxide reduction. The pathway involves activation of acetate to acetyl-SCoA followed by decarbonylation and reduction of the methyl group to methane coupled to the oxidation of the carbonyl group to carbon dioxide.

  9. Bioactive fractions from the pasture legume Biserrula pelecinus L. have an anti-methanogenic effect against key rumen methanogens.

    PubMed

    Banik, Bidhyut K; Durmic, Zoey; Erskine, William; Revell, Clinton K; Vadhanabhuti, Joy; McSweeney, Christopher S; Padmanabha, Jagadish; Flematti, Gavin R; Algreiby, Azizah A; Vercoe, Philip E

    2016-06-01

    Methanogenic archaea (methanogens) are common inhabitants of the mammalian intestinal tract. In ruminants, they are responsible for producing abundant amounts of methane during digestion of food, but selected bioactive plants and compounds may inhibit this activity. Recently, we have identified that, Biserrula pelecinus L. (biserrula) is one such plant and the current study investigated the specific anti-methanogenic activity of the plant. Bioassay-guided extraction and fractionation, coupled with in vitro fermentation batch culture were used to select the most bioactive fractions of biserrula. The four fractions were then tested against five species of methanogens grown in pure culture. Fraction bioactivity was assessed by measuring methane production and amplification of the methanogen mcrA gene. Treatments that showed bioactivity were subcultured in fresh broth without the bioactive fraction to distinguish between static and cidal effects. All four fractions were active against pure cultures, but the F2 fraction was the most consistent inhibitor of both methane production and cell growth, affecting four species of methanogens and also producing equivocal-cidal effects on the methanogens. Other fractions had selective activity affecting only some methanogens, or reducing either methane production or methanogenic cell growth. In conclusion, the anti-methanogenic activity of biserrula can be linked to compounds contained in selected bioactive fractions, with the F2 fraction strongly affecting key rumen methanogens. Further study is required to identify the specific plant compounds in biserrula that are responsible for the anti-methanogenic activity. These findings will help devise novel strategies to control methanogen populations and activity in the rumen, and consequently contribute in reducing greenhouse gas emissions from ruminants. PMID:27060275

  10. Methanogens: Methane Producers of the Rumen and Mitigation Strategies

    PubMed Central

    Hook, Sarah E.; Wright, André-Denis G.; McBride, Brian W.

    2010-01-01

    Methanogens are the only known microorganisms capable of methane production, making them of interest when investigating methane abatement strategies. A number of experiments have been conducted to study the methanogen population in the rumen of cattle and sheep, as well as the relationship that methanogens have with other microorganisms. The rumen methanogen species differ depending on diet and geographical location of the host, as does methanogenesis, which can be reduced by modifying dietary composition, or by supplementation of monensin, lipids, organic acids, or plant compounds within the diet. Other methane abatement strategies that have been investigated are defaunation and vaccines. These mitigation methods target the methanogen population of the rumen directly or indirectly, resulting in varying degrees of efficacy. This paper describes the methanogens identified in the rumens of cattle and sheep, as well as a number of methane mitigation strategies that have been effective in vivo. PMID:21253540

  11. High tolerance of methanogens in granular sludge to oxygen

    SciTech Connect

    Kato, M.T.; Field, J.A.; Lettinga, G. . Dept. of Environmental Technology)

    1993-12-01

    This research assessed the effect of oxygen exposure on the methanogenic activity of anaerobic granular sludges. The toxicity of oxygen to acetoclastic methanogens in five different anaerobic granular sludges was determined in serum flasks with effective gas-to-liquid volumes of 4.65 to 1. The amount of oxygen that caused 50% inhibition of the methanogenic activity after 3 days of exposure ranged from 7% to 41% oxygen in the head space. These results indicate that methanogens located in granular sludge have a high tolerance for oxygen. The most important factor contributing to the tolerance was the oxygen consumption by facultative bacteria metabolizing biodegradable substrates. Uptake of oxygen by these bacteria creates anaerobic microenvironments where the methanogenic bacteria are protected. The results also indicate that methanogens in sludge consortia still have some tolerance to oxygen, even in the absence of facultative substrate for oxygen respiration.

  12. Methanopyrus kandleri: an archaeal methanogen unrelated to all other known methanogens

    NASA Technical Reports Server (NTRS)

    Burggraf, S.; Stetter, K. O.; Rouviere, P.; Woese, C. R.

    1991-01-01

    Analysis of its 16S rRNA sequence shows that the newly discovered hyperthermophilic methanogen, Methanopryus kandleri, is phylogenetically unrelated to any other known methanogen. The organism represents a separate lineage originating near the root of the archaeal tree. Although the 16S rRNA sequence of Mp. kandleri resembles euryarchaeal 16S rRNAs more than it does crenarchaeal, it shows more crenarchaeal signature features than any known euryarchaeal rRNA. Attempts to place it in relation to the root of the archaeal tree show that the Mp. kandleri lineage likely arises from the euryarchaeal branch of the tree. While the existence of so deeply branching a methanogenic lineage brings into question the thesis that methanogenesis evolved from an earlier metabolism similar to that seen in Thermococcus, it at the same time reinforces the notion that the aboriginal [correction of aborginal] archaeon was a thermophile.

  13. Levels of water-soluble vitamins in methanogenic and non-methanogenic bacteria

    SciTech Connect

    Leigh, J.A.

    1983-03-01

    The levels of seven water-soluble vitamins in Methanobacterium thermoautotropicum, Methanococcus voltae, Escherichia coli, Bacillus subtillis, Pseudomonas fluorescens, and Bacteroides thetaiotaomicron were compared by using a vitamin-requiring Leuconostoc strain. Both methanogens contained levels of folic acid and pantothenic acid which were approximately two orders of magnitude lower than levels in the nonmethanogens. Methanobacterium thermoautotrophicum contained levels of thiamine, biotin, nicotinic acid, and pyridoxine which were approximately one order of magnitude lower than levels in the nonmethanogens. The thiamine level in Methanococcus voltae was approximately one order of magnitude lower than levels in the nonmethanogens. Only the levels of riboflavin (and nicotinic acid and pyridoxine in Methanococcus voltae) were approximately equal in the methanogens and nonmethanogens. Folic acid may have been present in extracts of methanogens merely as a precursor, by-product, or hydrolysis product of methanopterin.

  14. Glycine betaine as a direct substrate for methanogens (Methanococcoides spp.).

    PubMed

    Watkins, Andrew J; Roussel, Erwan G; Parkes, R John; Sass, Henrik

    2014-01-01

    Nine marine methanogenic Methanococcoides strains, including the type strains of Methanococcoides methylutens, M. burtonii, and M. alaskense, were tested for the utilization of N-methylated glycines. Three strains (NM1, PM2, and MKM1) used glycine betaine (N,N,N-trimethylglycine) as a substrate for methanogenesis, partially demethylating it to N,N-dimethylglycine, whereas none of the strains used N,N-dimethylglycine or sarcosine (N-methylglycine). Growth rates and growth yields per mole of substrate with glycine betaine (3.96 g [dry weight] per mol) were similar to those with trimethylamine (4.11 g [dry weight] per mol). However, as glycine betaine is only partially demethylated, the yield per methyl group was significantly higher than with trimethylamine. If glycine betaine and trimethylamine are provided together, trimethylamine is demethylated to dimethyl- and methylamine with limited glycine betaine utilization. After trimethylamine is depleted, dimethylamine and glycine betaine are consumed rapidly, before methylamine. Glycine betaine extends the range of substrates that can be directly utilized by some methanogens, allowing them to gain energy from the substrate without the need for syntrophic partners. PMID:24162571

  15. Community and Proteomic Analysis of Methanogenic Consortia Degrading Terephthalate

    PubMed Central

    Wu, Feng-Yau; Chuang, Hui-Ping; Chen, Wei-Yu; Huang, Hung-Jen; Chen, Shu-Hui; Liu, Wen-Tso

    2013-01-01

    Degradation of terephthalate (TA) through microbial syntrophy under moderately thermophilic (46 to 50°C) methanogenic conditions was characterized by using a metagenomic approach (A. Lykidis et al., ISME J. 5:122–130, 2011). To further study the activities of key microorganisms responsible for the TA degradation, community analysis and shotgun proteomics were used. The results of hierarchical oligonucleotide primer extension analysis of PCR-amplified 16S rRNA genes indicated that Pelotomaculum, Methanosaeta, and Methanolinea were predominant in the TA-degrading biofilms. Metaproteomic analysis identified a total of 482 proteins and revealed a distinctive distribution pattern of microbial functions expressed in situ. The results confirmed that TA was degraded by Pelotomaculum spp. via the proposed decarboxylation and benzoyl-coenzyme A-dependent pathway. The intermediate by-products, including acetate, H2/CO2, and butyrate, were produced to support the growth of methanogens, as well as other microbial populations that could further degrade butyrate. Proteins related to energy production and conservation, and signal transduction mechanisms (that is, chemotaxis, PAS/GGDEF regulators, and stress proteins) were highly expressed, and these mechanisms were important for growth in energy-limited syntrophic ecosystems. PMID:23064332

  16. Glycine Betaine as a Direct Substrate for Methanogens (Methanococcoides spp.)

    PubMed Central

    Watkins, Andrew J.; Roussel, Erwan G.; Parkes, R. John

    2014-01-01

    Nine marine methanogenic Methanococcoides strains, including the type strains of Methanococcoides methylutens, M. burtonii, and M. alaskense, were tested for the utilization of N-methylated glycines. Three strains (NM1, PM2, and MKM1) used glycine betaine (N,N,N-trimethylglycine) as a substrate for methanogenesis, partially demethylating it to N,N-dimethylglycine, whereas none of the strains used N,N-dimethylglycine or sarcosine (N-methylglycine). Growth rates and growth yields per mole of substrate with glycine betaine (3.96 g [dry weight] per mol) were similar to those with trimethylamine (4.11 g [dry weight] per mol). However, as glycine betaine is only partially demethylated, the yield per methyl group was significantly higher than with trimethylamine. If glycine betaine and trimethylamine are provided together, trimethylamine is demethylated to dimethyl- and methylamine with limited glycine betaine utilization. After trimethylamine is depleted, dimethylamine and glycine betaine are consumed rapidly, before methylamine. Glycine betaine extends the range of substrates that can be directly utilized by some methanogens, allowing them to gain energy from the substrate without the need for syntrophic partners. PMID:24162571

  17. Study of methanogen communities associated with different rumen protozoal populations

    PubMed Central

    Belanche, Alejandro; de la Fuente, Gabriel; Newbold, Charles J

    2014-01-01

    Protozoa-associated methanogens (PAM) are considered one of the most active communities in the rumen methanogenesis. This experiment investigated whether methanogens are sequestrated within rumen protozoa, and structural differences between rumen free-living methanogens and PAM. Rumen protozoa were harvested from totally faunated sheep, and six protozoal fractions (plus free-living microorganisms) were generated by sequential filtration. Holotrich-monofaunated sheep were also used to investigate the holotrich-associated methanogens. Protozoal size determined the number of PAM as big protozoa had 1.7–3.3 times more methanogen DNA than smaller protozoa, but also more endosymbiotic bacteria (2.2- to 3.5-fold times). Thus, similar abundance of methanogens with respect to total bacteria were observed across all protozoal fractions and free-living microorganisms, suggesting that methanogens are not accumulated within rumen protozoa in a greater proportion to that observed in the rumen as a whole. All rumen methanogen communities had similar diversity (22.2 ± 3.4 TRFs). Free-living methanogens composed a conserved community (67% similarity within treatment) in the rumen with similar diversity but different structures than PAM (P < 0.05). On the contrary, PAM constituted a more variable community (48% similarity), which differed between holotrich and total protozoa (P < 0.001). Thus, PAM constitutes a community, which requires further investigation as part of methane mitigation strategies. PMID:25195951

  18. Molecular Biology and Genetics of the Acetate-Utilizing Methanogenic Bacteria

    SciTech Connect

    Robert P. Gunsalus

    2003-07-21

    Methane biosynthesis by the Methanosarcina species, in contrast to other methanogens, occurs from the full range of methanogenic substrates that include acetate, methanol, tri-methyl, di-methyl, and methyl-amine, methyl-sulfides, and in limited instances, H2/CO2. The Methanosarcina are also versatile in their ability to adapt and grow in habitats of varying osmolarity ranging from fresh water environments, marine environments, and to hyper saline environments (ca to 1.2 M NaCl). To facilitate studies that address the biochemistry, molecular biology and physiology of these organisms, we have constructed a whole-genome microarray to identify classes of differentially expressed genes in M. mazei strain Goe1. We propose to further identify and examine how genes and their proteins involved in the synthesis and transport of osmolytes in the cell are regulated. These compounds include N-epsilon-acetyl-beta-lysine, alpha-glutamate, betaine, and potassium whose levels within the cell are modulated in order to provide appropriate osmotic balance. We will identify differentially expressed genes involved in hydrogen and carbon dioxide sequestration since M. mazei strain Goe1 is currently the only practical model for such study. Finally, we will explore the essential roles of two metals, molybdate and tungstate, in methanogen regulation and metabolism of these environmentally essential organsims. The above studies will advance our general understanding of how methanogens respond to their environmental signals, and adapt by adjusting their physiology to thrive in changing anaerobic habitats whether natural or man-made.

  19. [Influence of Temperature on the Anaerobic Packed Bed Reactor Performance and Methanogenic Community].

    PubMed

    Xie, Hai-ying; Wang, Xin; Li, Mu-yuan; Yan, Xu-you; Igarashi, Yasuo; Luo, Feng

    2015-11-01

    This study aimed to analyze the effect of temperature on performance and microbial community structure of an anaerobic packed bed reactor (APBR). The temperature was increased step-wise from room temperature (22 degrees C ± 1 degrees C) to psychrophilic (15 degrees C ± 1 degrees C), mesophilic (37 degrees C ± 1 degrees C) and thermophilic (55 degrees C ± 1 degrees C). The results showed that, in the temperature changing process, the higher the temperature of APBR was, the higher COD removal rate and daily gas production were. After temperature changed to psychrophilic, mesophilic and thermophilic, COD removal rate and daily gas production were 25%, 45%, 60% and 2.3 L x d(-1), 4.0 L x d(-1), 8.5 L x d(-1) respectively. However, there was no significant change in biogas composition (-60%). A sudden temperature change caused a simultaneous increase in the concentration of volatile fatty acids (VFA), which had been fluctuating. Using 16S rRNA gene clone library screening, Euryarchaeota was commonly found, including important methanogens: MBT (Methanobacteriales), Mst (Methanosaetaceae) , Msc (Methanosarcinaceae) and MMB (Methanomicrobiales), as well as thermophilic bacteria and few spring Archaea. However, the diversity of methanogenic groups was reduced, especially at mesophilic. The results of quantitative PCR showed that the 16S rRNA gene concentrations of Mst, MMB and Msc were reduced by temperature changes. Although the relative proportion of every kind of methanogen was significantly affected, Mst was the dominant methanogen. PMID:26911011

  20. Decreasing ammonia inhibition in thermophilic methanogenic bioreactors using carbon fiber textiles.

    PubMed

    Sasaki, Kengo; Morita, Masahiko; Hirano, Shin-ichi; Ohmura, Naoya; Igarashi, Yasuo

    2011-05-01

    Ammonia accumulation is one of the main causes of the loss of methane production observed during fermentation. We investigated the effect of addition of carbon fiber textiles (CFT) to thermophilic methanogenic bioreactors with respect to ammonia tolerance during the process of degradation of artificial garbage slurry, by comparing the performance of the reactors containing CFT with the performance of reactors without CFT. Under total ammonia-N concentrations of 3,000 mg L(-1), the reactors containing CFT were found to mediate stable removal of organic compounds and methane production. Under these conditions, high levels of methanogenic archaea were retained at the CFT, as determined by 16S rRNA gene analysis for methanogenic archaea. In addition, Methanobacterium sp. was found to be dominant in the suspended fraction, and Methanosarcina sp. was dominant in the retained fraction of the reactors with CFT. However, the reactors without CFT had lower rates of removal of organic compounds and production of methane under total ammonia-N concentrations of 1,500 mg L(-1). Under this ammonia concentration, a significant accumulation of acetate was observed in the reactors without CFT (130.0 mM), relative to the reactors with CFT (4.2 mM). Only Methanobacterium sp. was identified in the reactors without CFT. These results suggest that CFT enables stable proliferation of aceticlastic methanogens by preventing ammonia inhibition. This improves the process of stable garbage degradation and production of methane in thermophilic bioreactors that include high levels of ammonia. PMID:21468711

  1. Genomic Characterization of Methanomicrobiales Reveals Three Classes of Methanogens

    SciTech Connect

    Anderson, Iain; Ulrich, Luke; Lupa, Boguslaw; Susanti, Dwi; Porat, I.; Hooper, Sean; Lykidis, A; Sieprawska-Lupa, Magdalena; Dharmarajan, Lakshmi; Goltsman, Eugene; Lapidus, Alla L.; Saunders, Elizabeth H; Han, Cliff; Land, Miriam L; Lucas, Susan; Mukhopadhyay, Biswarup; Whitman, William; Woese, Carl; Bristow, James; Kyrpides, Nikos C

    2009-01-01

    Background Methanomicrobiales is the least studied order of methanogens. While these organisms appear to be more closely related to the Methanosarcinales in ribosomal-based phylogenetic analyses, they are metabolically more similar to Class I methanogens. Methodology/Principal Findings In order to improve our understanding of this lineage, we have completely sequenced the genomes of two members of this order, Methanocorpusculum labreanum Z and Methanoculleus marisnigri JR1, and compared them with the genome of a third, Methanospirillum hungatei JF-1. Similar to Class I methanogens, Methanomicrobiales use a partial reductive citric acid cycle for 2-oxoglutarate biosynthesis, and they have the Eha energy-converting hydrogenase. In common with Methanosarcinales, Methanomicrobiales possess the Ech hydrogenase and at least some of them may couple formylmethanofuran formation and heterodisulfide reduction to transmembrane ion gradients. Uniquely, M. labreanum and M. hungatei contain hydrogenases similar to the Pyrococcus furiosus Mbh hydrogenase, and all three Methanomicrobiales have anti-sigma factor and anti-anti-sigma factor regulatory proteins not found in other methanogens. Phylogenetic analysis based on seven core proteins of methanogenesis and cofactor biosynthesis places the Methanomicrobiales equidistant from Class I methanogens and Methanosarcinales. Conclusions/Significance Our results indicate that Methanomicrobiales, rather than being similar to Class I methanogens or Methanomicrobiales, share some features of both and have some unique properties. We find that there are three distinct classes of methanogens: the Class I methanogens, the Methanomicrobiales (Class II), and the Methanosarcinales (Class III).

  2. Genomic Characterization of Methanomicrobiales Reveals Three Classes of Methanogens

    SciTech Connect

    Anderson, Iain; Ulrich, Luke E.; Lupa, Boguslaw; Susanti, Dwi; Porat, Iris; Hooper, Sean D.; Lykidis, Athanasios; Sieprawska-Lupa, Magdalena; Dharmarajan, Lakshmi; Goltsman, Eugene; Lapidus, Alla; Saunders, Elizabeth; Han, Cliff; Land, Miriam; Lucas, Susan; Mukhopadhyay, Biswarup; Whitman, William B.; Woese, Carl; Bristow, James; Kyrpides, Nikos

    2009-05-01

    Methanomicrobiales is the least studied order of methanogens. While these organisms appear to be more closely related to the Methanosarcinales in ribosomal-based phylogenetic analyses, they are metabolically more similar to Class I methanogens. In order to improve our understanding of this lineage, we have completely sequenced the genomes of two members of this order, Methanocorpusculum labreanum Z and Methanoculleus marisnigri JR1, and compared them with the genome of a third, Methanospirillum hungatei JF-1. Similar to Class I methanogens, Methanomicrobiales use a partial reductive citric acid cycle for 2-oxoglutarate biosynthesis, and they have the Eha energy-converting hydrogenase. In common with Methanosarcinales, Methanomicrobiales possess the Ech hydrogenase and at least some of them may couple formylmethanofuran formation and heterodisulfide reduction to transmembrane ion gradients. Uniquely, M. labreanum and M. hungatei contain hydrogenases similar to the Pyrococcus furiosus Mbh hydrogenase, and all three Methanomicrobiales have anti-sigma factor and anti-anti-sigma factor regulatory proteins not found in other methanogens. Phylogenetic analysis based on seven core proteins of methanogenesis and cofactor biosynthesis places the Methanomicrobiales equidistant from Class I methanogens and Methanosarcinales. Our results indicate that Methanomicrobiales, rather than being similar to Class I methanogens or Methanomicrobiales, share some features of both and have some unique properties. We find that there are three distinct classes of methanogens: the Class I methanogens, the Methanomicrobiales (Class II), and the Methanosarcinales (Class III).

  3. A 1,681-locus consensus genetic map of cultivated cucumber including 67 NB-LRR resistance gene homolog and ten gene loci

    PubMed Central

    2013-01-01

    Background Cucumber is an important vegetable crop that is susceptible to many pathogens, but no disease resistance (R) genes have been cloned. The availability of whole genome sequences provides an excellent opportunity for systematic identification and characterization of the nucleotide binding and leucine-rich repeat (NB-LRR) type R gene homolog (RGH) sequences in the genome. Cucumber has a very narrow genetic base making it difficult to construct high-density genetic maps. Development of a consensus map by synthesizing information from multiple segregating populations is a method of choice to increase marker density. As such, the objectives of the present study were to identify and characterize NB-LRR type RGHs, and to develop a high-density, integrated cucumber genetic-physical map anchored with RGH loci. Results From the Gy14 draft genome, 70 NB-containing RGHs were identified and characterized. Most RGHs were in clusters with uneven distribution across seven chromosomes. In silico analysis indicated that all 70 RGHs had EST support for gene expression. Phylogenetic analysis classified 58 RGHs into two clades: CNL and TNL. Comparative analysis revealed high-degree sequence homology and synteny in chromosomal locations of these RGH members between the cucumber and melon genomes. Fifty-four molecular markers were developed to delimit 67 of the 70 RGHs, which were integrated into a genetic map through linkage analysis. A 1,681-locus cucumber consensus map including 10 gene loci and spanning 730.0 cM in seven linkage groups was developed by integrating three component maps with a bin-mapping strategy. Physically, 308 scaffolds with 193.2 Mbp total DNA sequences were anchored onto this consensus map that covered 52.6% of the 367 Mbp cucumber genome. Conclusions Cucumber contains relatively few NB-LRR RGHs that are clustered and unevenly distributed in the genome. All RGHs seem to be transcribed and shared significant sequence homology and synteny with the melon

  4. Energetics of syntrophic cooperation in methanogenic degradation.

    PubMed Central

    Schink, B

    1997-01-01

    Fatty acids and alcohols are key intermediates in the methanogenic degradation of organic matter, e.g., in anaerobic sewage sludge digestors or freshwater lake sediments. They are produced by classical fermenting bacteria for disposal of electrons derived in simultaneous substrate oxidations. Methanogenic bacteria can degrade primarily only one-carbon compounds. Therefore, acetate, propionate, ethanol, and their higher homologs have to be fermented further to one-carbon compounds. These fermentations are called secondary or syntrophic fermentations. They are endergonic processes under standard conditions and depend on intimate coupling with methanogenesis. The energetic situation of the prokaryotes cooperating in these processes is problematic: the free energy available in the reactions for total conversion of substrate to methane attributes to each partner amounts of energy in the range of the minimum biochemically convertible energy, i.e., 20 to 25 kJ per mol per reaction. This amount corresponds to one-third of an ATP unit and is equivalent to the energy required for a monovalent ion to cross the charged cytoplasmic membrane. Recent studies have revealed that syntrophically fermenting bacteria synthesize ATP by substrate-level phosphorylation and reinvest part of the ATP-bound energy into reversed electron transport processes, to release the electrons at a redox level accessible by the partner bacteria and to balance their energy budget. These findings allow us to understand the energy economy of these bacteria on the basis of concepts derived from the bioenergetics of other microorganisms. PMID:9184013

  5. Comparative analysis of metagenomes from three methanogenic hydrocarbon-degrading enrichment cultures with 41 environmental samples.

    PubMed

    Tan, Boonfei; Fowler, S Jane; Abu Laban, Nidal; Dong, Xiaoli; Sensen, Christoph W; Foght, Julia; Gieg, Lisa M

    2015-09-01

    Methanogenic hydrocarbon metabolism is a key process in subsurface oil reservoirs and hydrocarbon-contaminated environments and thus warrants greater understanding to improve current technologies for fossil fuel extraction and bioremediation. In this study, three hydrocarbon-degrading methanogenic cultures established from two geographically distinct environments and incubated with different hydrocarbon substrates (added as single hydrocarbons or as mixtures) were subjected to metagenomic and 16S rRNA gene pyrosequencing to test whether these differences affect the genetic potential and composition of the communities. Enrichment of different putative hydrocarbon-degrading bacteria in each culture appeared to be substrate dependent, though all cultures contained both acetate- and H2-utilizing methanogens. Despite differing hydrocarbon substrates and inoculum sources, all three cultures harbored genes for hydrocarbon activation by fumarate addition (bssA, assA, nmsA) and carboxylation (abcA, ancA), along with those for associated downstream pathways (bbs, bcr, bam), though the cultures incubated with hydrocarbon mixtures contained a broader diversity of fumarate addition genes. A comparative metagenomic analysis of the three cultures showed that they were functionally redundant despite their enrichment backgrounds, sharing multiple features associated with syntrophic hydrocarbon conversion to methane. In addition, a comparative analysis of the culture metagenomes with those of 41 environmental samples (containing varying proportions of methanogens) showed that the three cultures were functionally most similar to each other but distinct from other environments, including hydrocarbon-impacted environments (for example, oil sands tailings ponds and oil-affected marine sediments). This study provides a basis for understanding key functions and environmental selection in methanogenic hydrocarbon-associated communities. PMID:25734684

  6. Comparative analysis of metagenomes from three methanogenic hydrocarbon-degrading enrichment cultures with 41 environmental samples

    PubMed Central

    Tan, Boonfei; Jane Fowler, S; Laban, Nidal Abu; Dong, Xiaoli; Sensen, Christoph W; Foght, Julia; Gieg, Lisa M

    2015-01-01

    Methanogenic hydrocarbon metabolism is a key process in subsurface oil reservoirs and hydrocarbon-contaminated environments and thus warrants greater understanding to improve current technologies for fossil fuel extraction and bioremediation. In this study, three hydrocarbon-degrading methanogenic cultures established from two geographically distinct environments and incubated with different hydrocarbon substrates (added as single hydrocarbons or as mixtures) were subjected to metagenomic and 16S rRNA gene pyrosequencing to test whether these differences affect the genetic potential and composition of the communities. Enrichment of different putative hydrocarbon-degrading bacteria in each culture appeared to be substrate dependent, though all cultures contained both acetate- and H2-utilizing methanogens. Despite differing hydrocarbon substrates and inoculum sources, all three cultures harbored genes for hydrocarbon activation by fumarate addition (bssA, assA, nmsA) and carboxylation (abcA, ancA), along with those for associated downstream pathways (bbs, bcr, bam), though the cultures incubated with hydrocarbon mixtures contained a broader diversity of fumarate addition genes. A comparative metagenomic analysis of the three cultures showed that they were functionally redundant despite their enrichment backgrounds, sharing multiple features associated with syntrophic hydrocarbon conversion to methane. In addition, a comparative analysis of the culture metagenomes with those of 41 environmental samples (containing varying proportions of methanogens) showed that the three cultures were functionally most similar to each other but distinct from other environments, including hydrocarbon-impacted environments (for example, oil sands tailings ponds and oil-affected marine sediments). This study provides a basis for understanding key functions and environmental selection in methanogenic hydrocarbon-associated communities. PMID:25734684

  7. Hydrogen-limited growth of hyperthermophilic methanogens at deep-sea hydrothermal vents

    PubMed Central

    Ver Eecke, Helene C.; Butterfield, David A.; Huber, Julie A.; Lilley, Marvin D.; Olson, Eric J.; Roe, Kevin K.; Evans, Leigh J.; Merkel, Alexandr Y.; Cantin, Holly V.; Holden, James F.

    2012-01-01

    Microbial productivity at hydrothermal vents is among the highest found anywhere in the deep ocean, but constraints on microbial growth and metabolism at vents are lacking. We used a combination of cultivation, molecular, and geochemical tools to verify pure culture H2 threshold measurements for hyperthermophilic methanogenesis in low-temperature hydrothermal fluids from Axial Volcano and Endeavour Segment in the northeastern Pacific Ocean. Two Methanocaldococcus strains from Axial and Methanocaldococcus jannaschii showed similar Monod growth kinetics when grown in a bioreactor at varying H2 concentrations. Their H2 half-saturation value was 66 μM, and growth ceased below 17–23 μM H2, 10-fold lower than previously predicted. By comparison, measured H2 and CH4 concentrations in fluids suggest that there was generally sufficient H2 for Methanocaldococcus growth at Axial but not at Endeavour. Fluids from one vent at Axial (Marker 113) had anomalously high CH4 concentrations and contained various thermal classes of methanogens based on cultivation and mcrA/mrtA analyses. At Endeavour, methanogens were largely undetectable in fluid samples based on cultivation and molecular screens, although abundances of hyperthermophilic heterotrophs were relatively high. Where present, Methanocaldococcus genes were the predominant mcrA/mrtA sequences recovered and comprised ∼0.2–6% of the total archaeal community. Field and coculture data suggest that H2 limitation may be partly ameliorated by H2 syntrophy with hyperthermophilic heterotrophs. These data support our estimated H2 threshold for hyperthermophilic methanogenesis at vents and highlight the need for coupled laboratory and field measurements to constrain microbial distribution and biogeochemical impacts in the deep sea. PMID:22869718

  8. Hydrogen-limited growth of hyperthermophilic methanogens at deep-sea hydrothermal vents.

    PubMed

    Ver Eecke, Helene C; Butterfield, David A; Huber, Julie A; Lilley, Marvin D; Olson, Eric J; Roe, Kevin K; Evans, Leigh J; Merkel, Alexandr Y; Cantin, Holly V; Holden, James F

    2012-08-21

    Microbial productivity at hydrothermal vents is among the highest found anywhere in the deep ocean, but constraints on microbial growth and metabolism at vents are lacking. We used a combination of cultivation, molecular, and geochemical tools to verify pure culture H(2) threshold measurements for hyperthermophilic methanogenesis in low-temperature hydrothermal fluids from Axial Volcano and Endeavour Segment in the northeastern Pacific Ocean. Two Methanocaldococcus strains from Axial and Methanocaldococcus jannaschii showed similar Monod growth kinetics when grown in a bioreactor at varying H(2) concentrations. Their H(2) half-saturation value was 66 μM, and growth ceased below 17-23 μM H(2), 10-fold lower than previously predicted. By comparison, measured H(2) and CH(4) concentrations in fluids suggest that there was generally sufficient H(2) for Methanocaldococcus growth at Axial but not at Endeavour. Fluids from one vent at Axial (Marker 113) had anomalously high CH(4) concentrations and contained various thermal classes of methanogens based on cultivation and mcrA/mrtA analyses. At Endeavour, methanogens were largely undetectable in fluid samples based on cultivation and molecular screens, although abundances of hyperthermophilic heterotrophs were relatively high. Where present, Methanocaldococcus genes were the predominant mcrA/mrtA sequences recovered and comprised ∼0.2-6% of the total archaeal community. Field and coculture data suggest that H(2) limitation may be partly ameliorated by H(2) syntrophy with hyperthermophilic heterotrophs. These data support our estimated H(2) threshold for hyperthermophilic methanogenesis at vents and highlight the need for coupled laboratory and field measurements to constrain microbial distribution and biogeochemical impacts in the deep sea. PMID:22869718

  9. Application of the fluorescent-antibody technique to the study of a methanogenic bacterium in lake sediments.

    PubMed

    Strayer, R F; Tiedje, J M

    1978-01-01

    Fluorescent antibody (FA) was prepared for a methanogenic bacterium isolated from Wintergreen Lake pelagic sediment. The isolate resembles Methanobacterium formicicum. The FA did not cross-react with 9 other methanogens, including M. formicicum strains, or 24 heterotrophs, 18 of which had been isolated from Wintergreen Lake sediment. FA-reacting methanogens were detected in heat-fixed smears of several different lake sediments and anaerobic sewage sludge. Pretreatment of all samples with either rhodamine-conjugated geletin or bovine serum albumin adequately controlled nonspecific absorption of the FA. Autofluorescent particles were observed in the sediment samples but, with experience, they could easily be distinguished from FA-reacting bacteria. FA direct counts of the specific methanogen in Wintergreen Lake sediments were made on four different sampling dates and compared with five-tube most-probable-number estimates of the total methanogenic population that was present in the same samples. The FA counts ranged from 3.1 X 10(6) to 1.4 X 10(7)/g of dry sediment. The highest most-probable-number estimates were at least an order ofmagnitude lower. PMID:341807

  10. Application of the fluorescent-antibody technique to the study of a methanogenic bacterium in lake sediments.

    PubMed Central

    Strayer, R F; Tiedje, J M

    1978-01-01

    Fluorescent antibody (FA) was prepared for a methanogenic bacterium isolated from Wintergreen Lake pelagic sediment. The isolate resembles Methanobacterium formicicum. The FA did not cross-react with 9 other methanogens, including M. formicicum strains, or 24 heterotrophs, 18 of which had been isolated from Wintergreen Lake sediment. FA-reacting methanogens were detected in heat-fixed smears of several different lake sediments and anaerobic sewage sludge. Pretreatment of all samples with either rhodamine-conjugated geletin or bovine serum albumin adequately controlled nonspecific absorption of the FA. Autofluorescent particles were observed in the sediment samples but, with experience, they could easily be distinguished from FA-reacting bacteria. FA direct counts of the specific methanogen in Wintergreen Lake sediments were made on four different sampling dates and compared with five-tube most-probable-number estimates of the total methanogenic population that was present in the same samples. The FA counts ranged from 3.1 X 10(6) to 1.4 X 10(7)/g of dry sediment. The highest most-probable-number estimates were at least an order ofmagnitude lower. Images PMID:341807

  11. Effects of soil structure destruction on methane production andcarbon partitioning between methanogenic pathways in tropical rain forestsoils

    SciTech Connect

    Teh, Yit Arn; Silver, Whendee L.

    2005-01-25

    Controls on methanogenesis are often determined fromlaboratory incubations of soils converted to slurries. Destruction ofsoil structure during slurry conversion may disrupt syntrophicassociations, kill methanogens, and/or alter the microsite distributionof methanogenic activity, suppressing CH4 production. The effects ofslurry conversion on methanogenesis were investigated to determine ifdisruption of aggregate structure impacted methanogenesis, substrateutilization, and C partitioning between methanogenic pathways. Soils werecollected from the tropical rain forest life zone of the LuquilloExperimental Forest, Puerto Rico, and exposed to different physicaldisturbances, including flooding and physical homogenization. Slurryconversion negatively impacted methanogenesis. Rates of CH4 productiondeclined by a factor of 17 after well-aggregated soils were converted toslurries. Significantly more 13C-acetate was recovered in CO2 compared toCH4 after slurry conversion, suggesting that methanogens consumed lessacetate after slurry conversion and may have competed less effectivelywith other anaerobes for acetate. Isotopic data indicate that therelative partitioning of C between aceticlastic and hydrogenotrophicpathways wasunchanged after slurry conversion. These data suggest thatexperiments which destroy soil structure may significantly underestimatemethanogenesis and overestimate the potential for other microorganisms tocompete with methanogens for organic substrates. Current knowledge of thefactors that regulate methanogenesis in soil may be biased by thefindings of slurry-based experiments, that do not accurately representthe complex, spatially heterogeneous conditions found in well-aggregatedsoils.

  12. [Community Structure and Succession of Methanogens in Beishenshu Landfill, Beijing].

    PubMed

    Song, Li-na; Wang, Lei; Xia, Meng-jing; Su, Yue; Li, Zhen-shan

    2015-09-01

    Methanogens are the key microorganisms for landfill stabilization. RT-PCR and qPCR detecting system were employed to determine the types and abundance of methanogens in 2-15 year-old solid wastes that sampled from Beishenshu Landfill, Beijing. The organic components were almost stable and the pH values were in alkaline range, which indicated that the landfill was in the methanogenic process. Methanobacterials, Methanosaeta, and Methanosarcina were detected, among which Methanosaeta and Methanosarcina are acetoclastic, and Methanobacterials are hydrogenotrophic. As landfill processing, within this time range, although the bacterial abundance was significantly decreased, the amount of methanogens was first increased and then decreased, and finally became stable after being landfilled for 9 years. Methanosarcina was the dominate taxa. Significant correlations were found between the methanogens and the volatile fatty acids, but the correlations between methanogens and larger molecular organic matters were relatively weak or even absent. Taken together, our study revealed that the amount of methanogens were affected by substrates, but hardly influenced by the conversion of large molecules in these wastes landfilled for more than 2 years. PMID:26717715

  13. Microbial precipitation of dolomite in methanogenic groundwater

    NASA Astrophysics Data System (ADS)

    Roberts, Jennifer A.; Bennett, Philip C.; González, Luis A.; MacPherson, G. L.; Milliken, Kitty L.

    2004-04-01

    We report low-temperature microbial precipitation of dolomite in dilute natural waters from both field and laboratory experiments. In a freshwater aquifer, microorganisms colonize basalt and nucleate nonstoichiometric dolomite on cell walls. In the laboratory, ordered dolomite formed at near-equilibrium conditions from groundwater with molar Mg:Ca ratios of <1; dolomite was absent in sterile experiments. Geochemical and microbiological data suggest that methanogens are the dominant metabolic guild in this system and are integral to dolomite precipitation. We hypothesize that the attached microbial consortium reacts with the basalt surface, releasing Mg and Ca into solution, which drives dolomite precipitation via nucleation on the cell wall. These findings provide insight into the long-standing dolomite problem and suggest a fundamental role for microbial processes in the formation of dolomite across a wide range of environmental conditions.

  14. Diversity of methanogens in the hindgut of captive white rhinoceroses, Ceratotherium simum

    PubMed Central

    2013-01-01

    Background The white rhinoceros is on the verge of extinction with less than 20,200 animals remaining in the wild. In order to better protect these endangered animals, it is necessary to better understand their digestive physiology and nutritional requirements. The gut microbiota is nutritionally important for herbivorous animals. However, little is known about the microbial diversity in the gastrointestinal tract (GIT) of the white rhinoceros. Methanogen diversity in the GIT may be host species-specific and, or, function-dependent. To assess methanogen diversity in the hindgut of white rhinoceroses, an archaeal 16S rRNA gene clone library was constructed from pooled PCR products obtained from the feces of seven adult animals. Results Sequence analysis of 153 archaeal 16S rRNA sequences revealed 47 unique phylotypes, which were assigned to seven operational taxonomic units (OTUs 1 to 7). Sequences assigned to OTU-7 (64 out of 153 total sequencs – 42%) and OTU-5 (18%, 27/153) had 96.2% and 95.5% identity to Methanocorpusculum labreanum, respectively, making Methanocorpusculum labreanum the predominant phylotype in these white rhynoceroses. Sequences belonging to OTU-6 (27%, 42/153) were related (97.6%) to Methanobrevibacter smithii. Only 4% of the total sequences (6/153) were assigned to Methanosphaera stadtmanae (OTU-1). Sequences belonging to OTU-2 (4%, 6/153), OTU-3 (3%, 5/153) and OTU-4 (2%, 3/153) were distantly related (87.5 to 88,4%) to Methanomassiliicoccus luminyensis and were considered to be novel species or strains that have yet-to-be cultivated and characterized. Conclusion Phylogenetic analysis indicated that the methanogen species in the hindgut of white rhinoceroses were more similar to those in the hindgut of horses. Our findings may help develop studies on improving the digestibility of forage for sustainable management and better health of these endangered animals. PMID:24228793

  15. Methanogenic Conversion of CO2 Into CH4

    SciTech Connect

    Stevens, S.H., Ferry, J.G., Schoell, M.

    2012-05-06

    This SBIR project evaluated the potential to remediate geologic CO2 sequestration sites into useful methane gas fields by application of methanogenic bacteria. Such methanogens are present in a wide variety of natural environments, converting CO2 into CH4 under natural conditions. We conclude that the process is generally feasible to apply within many of the proposed CO2 storage reservoir settings. However, extensive further basic R&D still is needed to define the precise species, environments, nutrient growth accelerants, and economics of the methanogenic process. Consequently, the study team does not recommend Phase III commercial application of the technology at this early phase.

  16. Methanogenic Community Dynamics during Anaerobic Utilization of Agricultural Wastes.

    PubMed

    Ziganshin, A M; Ziganshina, E E; Kleinsteuber, S; Pröter, J; Ilinskaya, O N

    2012-10-01

    This work is devoted to the investigation of the methanogenic archaea involved in anaerobic digestion of cattle manure and maize straw on the basis of terminal restriction fragment length polymorphism (T-RFLP) analysis of archaeal 16S rRNA genes. The biological diversity and dynamics of methanogenic communities leading to anaerobic degradation of agricultural organic wastes with biogas production were evaluated in laboratory-scale digesters. T-RFLP analysis, along with the establishment of archaeal 16S rRNA gene clone libraries, showed that the methanogenic consortium consisted mainly of members of the generaMethanosarcinaandMethanoculleus,with a predominance ofMethanosarcinaspp. throughout the experiment. PMID:23346384

  17. The role of hydrogenotrophic methanogens in an acidogenic reactor.

    PubMed

    Huang, Wenhai; Wang, Zhenyu; Zhou, Yan; Ng, Wun Jern

    2015-12-01

    A laboratory-scale acidogenic anaerobic sequencing batch reactor was set up to test the effect of pH change on microbial community structure of the reactor biomass and process performance. No immediate performance change on acidogenesis was observed after the pH change. However, as the hydrogenotrophic methanogen population decreased, hydrogen content in biogas increased followed by a sharp decrease in volatile fatty acids (VFAs) with acetic acid (HAc) in particular. Recovery of reactor performance following pH correction was only apparent after recovery of hydrogenotrophic methanogen population. These suggested hydrogenotrophic methanogens played a very important role in performance of the acidogenic process. PMID:25466820

  18. The potential of Methanogenic Life in the Solar System

    NASA Astrophysics Data System (ADS)

    Taubner, R.-S.; Firneis, M. G.; Leitner, J. J.; Schleper, C.; Rittmann, S. K.-M. R.

    2015-10-01

    Methanogens from the domain Archaea are obligate anaerobic chemolithoautotrophs or chemolithoheterotrophs producing methane (CH4). For the CH4-production they primarily use various C1 typecompounds (like carbon monoxide (CO), carbon dioxide (CO2), formate (HCO- 2)), but some strains are also able to utilize methanol (CH3OH), acetate, or even methylsulfides for energy production. The capability of methanogens thriving under various extreme environments on Earth is astonishing. Their enormous diversity and the similarity between their growth conditions and the environmental conditions on extraterrestrial bodies throughout the Solar System make methanogens to an ideal test object for astrobiological experiments.

  19. The Cultivated Classroom.

    ERIC Educational Resources Information Center

    Schilder, Rosalind

    1983-01-01

    Teachers who follow this monthly schedule for starting and cultivating plants in their classrooms can look forward to blooms and greenery throughout the year. Advice on choosing plants, making cuttings, forcing bulbs, rooting sweet potatoes and pineapples, and holding a Mother's Day plant sale is included. (PP)

  20. A hydrogen-based subsurface microbial community dominated by methanogens

    USGS Publications Warehouse

    Chapelle, F.H.; O'Neill, K.; Bradley, P.M.; Methe, B.A.; Ciufo, S.A.; Knobel, L.L.; Lovley, D.R.

    2002-01-01

    The search for extraterrestrial life may be facilitated if ecosystems can be found on Earth that exist under conditions analogous to those present on other planets or moons. It has been proposed, on the basis of geochemical and thermodynamic considerations, that geologically derived hydrogen might support subsurface microbial communities on Mars and Europa in which methanogens form the base of the ecosystem1-5. Here we describe a unique subsurface microbial community in which hydrogen-consuming, methane-producing Archaea far outnumber the Bacteria. More than 90% of the 16s ribosomal DNA sequences recovered from hydrothermal waters circulating through deeply buried igneous rocks in Idaho are related to hydrogen-using methanogenic microorganisms. Geochemical characterization indicates that geothermal hydrogen, not organic carbon, is the primary energy source for this methanogen-dominated microbial community. These results demonstrate that hydrogen-based methanogenic communities do occur in Earth's subsurface, providing an analogue for possible subsurface microbial ecosystems on other planets.

  1. Single-cell genome and metatranscriptome sequencing reveal metabolic interactions of an alkane-degrading methanogenic community

    PubMed Central

    Embree, Mallory; Nagarajan, Harish; Movahedi, Narjes; Chitsaz, Hamidreza; Zengler, Karsten

    2014-01-01

    Microbial interactions have a key role in global geochemical cycles. Although we possess significant knowledge about the general biochemical processes occurring in microbial communities, we are often unable to decipher key functions of individual microorganisms within the environment in part owing to the inability to cultivate or study them in isolation. Here, we circumvent this shortcoming through the use of single-cell genome sequencing and a novel low-input metatranscriptomics protocol to reveal the intricate metabolic capabilities and microbial interactions of an alkane-degrading methanogenic community. This methanogenic consortium oxidizes saturated hydrocarbons under anoxic conditions through a thus-far-uncharacterized biochemical process. The genome sequence of a dominant bacterial member of this community, belonging to the genus Smithella, was sequenced and served as the basis for subsequent analysis through metabolic reconstruction. Metatranscriptomic data generated from less than 500 pg of mRNA highlighted metabolically active genes during anaerobic alkane oxidation in comparison with growth on fatty acids. These data sets suggest that Smithella is not activating hexadecane by fumarate addition. Differential expression assisted in the identification of hypothetical proteins with no known homology that may be involved in hexadecane activation. Additionally, the combination of 16S rDNA sequence and metatranscriptomic data enabled the study of other prevalent organisms within the consortium and their interactions with Smithella, thus yielding a comprehensive characterization of individual constituents at the genome scale during methanogenic alkane oxidation. PMID:24152715

  2. Effect of salinity on methanogenic propionate degradation by acclimated marine sediment-derived culture.

    PubMed

    Miura, Toyokazu; Kita, Akihisa; Okamura, Yoshiko; Aki, Tsunehiro; Matsumura, Yukihiko; Tajima, Takahisa; Kato, Junichi; Nakashimada, Yutaka

    2015-12-01

    Degradation of propionate under high salinity is needed for biomethane production from salt-containing feedstocks. In this study, marine sediment-derived culture was evaluated to determine the effect of salinity on methanogenic propionate degradation. Microbes in marine sediments were subjected to fed-batch cultivation on propionate for developing acclimatized cultures. The rate of propionate degradation increased eightfold during 10 rounds of cultivation. Microbial community composition was determined through pyrosequencing of 16S rRNA gene amplicons after 10 rounds of cultivation. Taxa analysis was conducted for the reads obtained by pyrosequencing. Known propionate degraders were undetectable in the acclimated culture. Comparison of bacterial taxa in the original sediment with those in the acclimated culture revealed that the populations of four bacterial taxa were significantly increased during acclimation. Methanolobus was the predominant archaea genus in the acclimated culture. The propionate degradation rate of the acclimated culture was not affected by salinity of up to equivalent of 1.9 % NaCl. The rate decreased at higher salinity levels and was more than 50 % of the maximum rate even at equivalent of 4.3 % NaCl. PMID:26364311

  3. A comment on methanogenic bacteria and the primitive ecology

    NASA Technical Reports Server (NTRS)

    Woese, C. R.

    1977-01-01

    As the phenotype of methanogenic bacteria is suggested to have been one of the major factors creating a dynamic balance between CO2 and CH4 in the primitive atmosphere, these organisms are thought to be very ancient. Their antiquity may be further postulated by comparative characterization of their ribosomal RNA. Accepting this antiquity, it is concluded that a carbon-dioxide-methane cycle, driven by photosynthesis, was the major carbon cycle in primitive ecology, and that photosynthesis and methanogens were thus contemporaneous.

  4. Methanogen communities in a municipal landfill complex in China.

    PubMed

    Tang, Wei; Wang, Yangqing; Lei, Yu; Song, Liyan

    2016-05-01

    Landfills are significant global sources of atmospheric methane, but little is known about the ecology and community structure of methanogens in these sites. Here, we investigated the methanogen community based on methyl coenzyme M reductase A gene amplicons in the vertical profiles of three different sites at a municipal landfill complex in China. Links between methanogen communities and refuse properties were explored using multivariate analysis. Clone library results showed that most clones (92%) were related to the hydrogenotrophic methanogens, Methanomicrobiales. Almost all of the Methanomicrobiales clones retrieved in this study are members of the genus Methanoculleus Eight clones were affiliated with the genus Methanofollis The remaining clones were clustered within the genus Methanosarcina Terminal restriction fragment length polymorphism profiles showed that the landfill was predominated by 22 taxa, making up 69%-96% of the community. Of these, a single taxon comprised 36%-65% of the communities across all sites and depths. Principal components analysis separated the methanogen community into three groups, irrespective of site or depth. Redundancy analysis suggested that total phosphorus and pH play roles in structuring methanogen communities in landfills. PMID:27036145

  5. Evidence for para dechlorination of polychlorobiphenyls by methanogenic bacteria

    SciTech Connect

    Ye, D.; Quensen, J.F.; Tiedje, J.M.

    1995-06-01

    When microorganisms eluted from upper Hudson River sediment were cultured without any substrate except polychlorobiphenyl (PCB)-free Hudson River sediment, methane formation was the terminal step of the anaerobic food chain. In sediments containing Aroclor 1242, addition of eubacterium-inhibiting antibiotics, which should have directly inhibited fermentative bacteria and thereby should have indirectly inhibited methanogens, resulted in no dechlorination activity or methane production. However, when substrates for methanogenic bacteria were provided along with the antibiotics (to free the methanogens from dependence on eubacteria), concomitant methane production and dechlorination of PCBs were observed. The dechlorination of Aroclor 1242 was from the para positions, a pattern distinctly different from, and more limited than, the pattern observed with untreated or pasteurized inocula. Both methane production and dechlorination in cultures amended with antibiotics plus methanogenic substrates were inhibited by 2-bromoethanesulfonic acid. These results suggest that the methanogenic bacteria are among the physiological groups capable of anaerobic dechlorination of PCBs, but that the dechlorination observed with methanogenic bacteria is less extensive than the dechlorination observed with more complex anaerobic consortia. 27 refs., 5 figs., 1 tab.

  6. Methanosalsum natronophilum sp. nov., and Methanocalculus alkaliphilus sp. nov., haloalkaliphilic methanogens from hypersaline soda lakes.

    PubMed

    Sorokin, Dimitry Y; Abbas, Ben; Merkel, Alexander Y; Rijpstra, W Irene C; Damsté, Jaap S Sinninghe; Sukhacheva, Marina V; van Loosdrecht, Mark C M

    2015-10-01

    Two groups of haloalkaliphilic methanogenic archaea were dominating in enrichments from hypersaline soda lake sediments at pH 10. At moderate salt concentrations with formate or H2 as electron donor, methanogens belonging to the genus Methanocalculus were enriched, while at high salt concentrations with methylated substrates, a group related to Methanosalsum zhilinae was dominating. For both groups, several pure cultures were obtained including the type strains AMF2T for the Methanocalculus group and AME2T for the Methanosalsum group. The Methanocalculus group is characterized by lithoheterotrophic growth with either formate (preferable substrate) or H2 at moderate salinity up to 1.5-2 M total Na+ and obligate alkaliphilic growth with an optimum at pH 9.5. According to phylogenetic analysis, the group also includes closely related strains isolated previously from the low-salt alkaline Lonar Lake. The novel Methanosalsum group is characterized by high salt tolerance (up to 3.5 M total Na+) and obligate alkaliphilic growth with an optimum at pH 9.5. It has a typical methylotrophic substrate profile, utilizing methanol, methylamines and dimethyl sulfide (at low concentrations) as methanogenic substrates. On the basis of physiological and phylogenetic data, it is proposed that the two groups of soda lake methanogenic isolates are assigned into two novel species, Methanocalculus alkaliphilus sp. nov. (type strain AMF2T = DSM 24457T = UNIQEM U859T) and Methanosalsum natronophilum sp. nov. (type strain AME2T = DSM 24634T = NBRC 110091T). PMID:26228570

  7. Evaluation of methanogenic treatment of TMAH (tetra-methyl ammonium hydroxide) in a full-scale TFT-LCD wastewater treatment process.

    PubMed

    Hu, T H; Whang, L M; Lei, C N; Chen, C F; Chiang, T Y; Lin, L B; Chen, H W; Liu, P W G; Cheng, S S

    2010-01-01

    This study evaluated TMAH biodegradation under methanogenic conditions. Under methanogenic conditions, a sludge from a full-scale UASB treating TFT-LCD wastewater was able to degrade 2,000 mg/L of TMAH within 10 h and attained a specific degradation rate of 19.2 mgTMAH/gVSS-h. Furthermore, several chemicals including some surfactants, DMSO, and sulfate were examined for their potential inhibitory effects on TMAH biodegradation under methanogenic conditions. The results indicated that surfactant S1 (up to 2%) and DMSO (up to 1,000 mg/L) presented negligible inhibitory effects on TMAH degradation, while surfactant S2 (0.2-1%) might inhibit methanogenic reaction without any TMAH degradation for 3-5 h. At sulfate concentrations higher than 300 mg/L, a complete inhibition of methanogenic reaction and TMAH biodegradation was observed. Results from cloning and sequencing of archaeal 16S rRNA gene fragments showed that Methanosarcina barkeri and Methanosarcina mazei were the dominant methanogens in the UASB treating TMAH-containing TFT-LCD wastewater. PMID:20651446

  8. Dechlorination of polychlorinated methanes by a sequential methanogenic-denitrifying bioreactor system.

    PubMed

    Yu, Z; Smith, G B

    2000-04-01

    A two-stage bioreactor has been developed to link dechlorination of halogenated methane compounds to the anaerobic processes of methanogenesis and denitrification. A digester methanogenic consortium was shown to dechlorinate chloroform (CF) and carbon tetrachloride (CT) to dichloromethane (DCM), and DCM was then mineralized by an acclimated denitrifying biological activated carbon consortium. Combining these two processes, a sequential methanogenic-denitrifying bioreactor (SMDB) system that completely degraded polychlorinated methanes including CT, CF, and DCM was developed. More than 95% of the added CT and CF was dechlorinated in the methanogenic bioreactor with methanol as the primary substrate, and the resultant DCM was biodegraded in the denitrifying bioreactor with nitrate as the electron acceptor. In the denitrifying bioreactor, the residual CF was completely removed, and the DCM removal efficiency was more than 95%. This novel bioreactor system eliminates the need for aeration and so avoids the air contamination associated with aerobic biotreatment of volatile chlorinated pollutants. This SMDB system provides an alternative to conventional biotreatment of wastewaters and other matrices contaminated with polychlorinated methanes and is, to our knowledge, the first report on such a sequential anoxic system. PMID:10803908

  9. Terrestrial models for extraterrestrial life: methanogens and halophiles at Martian temperatures

    NASA Astrophysics Data System (ADS)

    Reid, I. N.; Sparks, W. B.; Lubow, S.; McGrath, M.; Livio, M.; Valenti, J.; Sowers, K. R.; Shukla, H. D.; MacAuley, S.; Miller, T.; Suvanasuthi, R.; Belas, R.; Colman, A.; Robb, F. T.; Dassarma, P.; Müller, J. A.; Coker, J. A.; Cavicchioli, R.; Chen, F.; Dassarma, S.

    2006-08-01

    Cold environments are common throughout the Galaxy. We are conducting a series of experiments designed to probe the low-temperature limits for growth in selected methanogenic and halophilic Archaea. This paper presents initial results for two mesophiles, a methanogen, Methanosarcina acetivorans, and a halophile, Halobacterium sp. NRC-1, and for two Antarctic cold-adapted Archaea, a methanogen, Methanococcoides burtonii, and a halophile, Halorubrum lacusprofundi. Neither mesophile is active at temperatures below 5 °C, but both cold-adapted microorganisms show significant growth at sub-zero temperatures (-2 °C and -1 °C, respectively), extending previous low-temperature limits for both species by 4 5 °C. At low temperatures, both H. lacusprofundi and M. burtonii form multicellular aggregates, which appear to be embedded in extracellular polymeric substances. This is the first detection of this phenomenon in Antarctic species of Archaea at cold temperatures. The low-temperature limits for both psychrophilic species fall within the temperature range experienced on present-day Mars and could permit survival and growth, particularly in sub-surface environments. We also discuss the results of our experiments in the context of known exoplanet systems, several of which include planets that intersect the Habitable Zone. In most cases, those planets follow orbits with significant eccentricity, leading to substantial temperature excursions. However, a handful of the known gas giant exoplanets could potentially harbour habitable terrestrial moons.

  10. Field Evidence for Magnetite Formation by a Methanogenic Microbial Community

    NASA Astrophysics Data System (ADS)

    Rossbach, S.; Beaver, C. L.; Williams, A.; Atekwana, E. A.; Slater, L. D.; Ntarlagiannis, D.; Lund, A.

    2015-12-01

    The aged, subsurface petroleum spill in Bemidji, Minnesota, has been surveyed with magnetic susceptibility (MS) measurements. High MS values were found in the free-product phase around the fluctuating water table. Although we had hypothesized that high MS values are related to the occurrence of the mineral magnetite resulting from the activity of iron-reducing bacteria, our microbial analysis pointed to the presence of a methanogenic microbial community at the locations and depths of the highest MS values. Here, we report on a more detailed microbial analysis based on high-throughput sequencing of the 16S rRNA gene of sediment samples from four consecutive years. In addition, we provide geochemical data (FeII/FeIII concentrations) to refine our conceptual model of methanogenic hydrocarbon degradation at aged petroleum spills and demonstrate that the microbial induced changes of sediment properties can be monitored with MS. The methanogenic microbial community at the Bemidji site consisted mainly of the syntrophic, hydrocarbon-degrading Smithella and the hydrogenotrophic, methane-generating Methanoregula. There is growing evidence in the literature that not only Bacteria, but also some methanogenic Archaea are able to reduce iron. In fact, a recent study reported that the methanogen Methanosarcina thermophila produced magnetite during the reduction of ferrihydrite in a laboratory experiment when hydrogen was present. Therefore, our finding of high MS values and the presence of magnetite in the methanogenic zone of an aged, subsurface petroleum spill could very well be the first field evidence for magnetite formation during methanogenic hydrocarbon degradation.

  11. Fate of neptunium in an anaerobic, methanogenic microcosm.

    SciTech Connect

    Banaszak, J. E.

    1998-12-21

    Neptunium is found predominantly as Np(IV) in reducing environments, but Np(V) in aerobic environments. However, currently it is not known how the interplay between biotic and abiotic processes affects Np redox speciation in the environment. In order to evaluate the effect of anaerobic microbial activity on the fate of Np in natural systems, Np(V) was added to a microcosminoculated with anaerobic sediments from a metal-contaminated fresh water lake. The consortium included metal-reducing, sulfate-reducing, and methanogenic microorganisms, and acetate was supplied as the only exogenous substrate. Addition of more than 10{sup {minus}5} M Np did not inhibit methane production. Total Np volubility in the active microcosm, as well as in sterilized control samples, decreased by nearly two orders of magnitude. A combination of analytical techniques, including VIS-NIR absorption spectroscopy and XANES, identified Np(IV) as the oxidation state associated with the sediments. The similar results from the active microcosm and the abiotic controls suggest that microbian y produced Mn(II/HI) and Fe(II) may serve as electron donors for Np reduction.

  12. Distribution of compatible solutes in the halophilic methanogenic archaebacteria.

    PubMed Central

    Lai, M C; Sowers, K R; Robertson, D E; Roberts, M F; Gunsalus, R P

    1991-01-01

    Accumulation of compatible solutes, by uptake or de novo synthesis, enables bacteria to reduce the difference between osmotic potentials of the cell cytoplasm and the extracellular environment. To examine this process in the halophilic and halotolerant methanogenic archaebacteria, 14 strains were tested for the accumulation of compatible solutes in response to growth in various extracellular concentrations of NaCl. In external NaCl concentrations of 0.7 to 3.4 M, the halophilic methanogens accumulated K+ ion and low-molecular-weight organic compounds. beta-Glutamate was detected in two halotolerant strains that grew below 1.5 M NaCl. Two unusual beta-amino acids, N epsilon-acetyl-beta-lysine and beta-glutamine (3-aminoglutaramic acid), as well as L-alpha-glutamate were compatible solutes among all of these strains. De novo synthesis of glycine betaine was also detected in several strains of moderately and extremely halophilic methanogens. The zwitterionic compounds (beta-glutamine, N epsilon-acetyl-beta-lysine, and glycine betaine) and potassium were the predominant compatible solutes among the moderately and extremely halophilic methanogens. This is the first report of beta-glutamine as a compatible solute and de novo biosynthesis of glycine betaine in the methanogenic archaebacteria. PMID:1909318

  13. Distribution of compatible solutes in the halophilic methanogenic archaebacteria

    SciTech Connect

    Meichin Lai; Sowers, K.R.; Gunsalus, R.P. ); Robertson, D.E.; Roberts, M.F. )

    1991-09-01

    Accumulation of compatible solutes, by uptake or de novo synthesis, enables bacteria to reduce the difference between osmotic potentials of the cell cytoplasm and the extracellular environment. To examine this process in the halophilic and halotolerant methanogenic archaebacteria, 14 strains were tested for the accumulation of compatible solutes in response to growth in various extracellular concentrations of NaCl. In external NaCl concentrations of 0.7 to 3.4 M, the halophilic methanogens accumulated K{sup +} ion and low-molecular-weight organic compounds. {beta}-Glutamate was detected in two halotolerant strains that grew below 1.5 M NaCl. Two unusual {beta}-amino acids, N{sub {var epsilon}}-acetyl-{beta}-lysine and {beta}-glutamine (3-aminoglutaramic acid), as well as L-{alpha}-glutamate were compatible solutes among all of these strains. De novo synthesis of glycine betaine was also detected in several strains of moderately and extremely halophilic methanogens. The zwitterionic compounds ({beta}-glutamine, N{sub {var epsilon}}-acetyl-{beta}-lysine,a nd glycine betaine) and potassium were the predominant compatible solutes among the moderately and extremely halophilic methanogens. This is the first report of {beta}-glutamine as a compatible solute and de novo biosynthesis of glycine betaine in the methanogenic archaebacteria.

  14. Methanogenic archaea isolated from Taiwan's Chelungpu fault.

    PubMed

    Wu, Sue-Yao; Lai, Mei-Chin

    2011-02-01

    Terrestrial rocks, petroleum reservoirs, faults, coal seams, and subseafloor gas hydrates contain an abundance of diverse methanoarchaea. However, reports on the isolation, purification, and characterization of methanoarchaea in the subsurface environment are rare. Currently, no studies investigating methanoarchaea within fault environments exist. In this report, we succeeded in obtaining two new methanogen isolates, St545Mb(T) of newly proposed species Methanolobus chelungpuianus and Methanobacterium palustre FG694aF, from the Chelungpu fault, which is the fault that caused a devastating earthquake in central Taiwan in 1999. Strain FG694aF was isolated from a fault gouge sample obtained at 694 m below land surface (mbls) and is an autotrophic, mesophilic, nonmotile, thin, filamentous-rod-shaped organism capable of using H(2)-CO(2) and formate as substrates for methanogenesis. The morphological, biochemical, and physiological characteristics and 16S rRNA gene sequence analysis revealed that this isolate belongs to Methanobacterium palustre. The mesophilic strain St545Mb(T), isolated from a sandstone sample at 545 mbls, is a nonmotile, irregular, coccoid organism that uses methanol and trimethylamine as substrates for methanogenesis. The 16S rRNA gene sequence of strain St545Mb(T) was 99.0% similar to that of Methanolobus psychrophilus strain R15 and was 96 to 97.5% similar to the those of other Methanolobus species. However, the optimal growth temperature and total cell protein profile of strain St545Mb(T) were different from those of M. psychrophilus strain R15, and whole-genome DNA-DNA hybridization revealed less than 20% relatedness between these two strains. On the basis of these observations, we propose that strain St545Mb(T) (DSM 19953(T); BCRC AR10030; JCM 15159) be named Methanolobus chelungpuianus sp. nov. Moreover, the environmental DNA database survey indicates that both Methanolobus chelungpuianus and Methanobacterium palustre are widespread in the

  15. Thermodynamics and H2 Transfer in a Methanogenic, Syntrophic Community

    PubMed Central

    Hamilton, Joshua J.; Calixto Contreras, Montserrat; Reed, Jennifer L.

    2015-01-01

    Microorganisms in nature do not exist in isolation but rather interact with other species in their environment. Some microbes interact via syntrophic associations, in which the metabolic by-products of one species serve as nutrients for another. These associations sustain a variety of natural communities, including those involved in methanogenesis. In anaerobic syntrophic communities, energy is transferred from one species to another, either through direct contact and exchange of electrons, or through small molecule diffusion. Thermodynamics plays an important role in governing these interactions, as the oxidation reactions carried out by the first community member are only possible because degradation products are consumed by the second community member. This work presents the development and analysis of genome-scale network reconstructions of the bacterium Syntrophobacter fumaroxidans and the methanogenic archaeon Methanospirillum hungatei. The models were used to verify proposed mechanisms of ATP production within each species. We then identified additional constraints and the cellular objective function required to match experimental observations. The thermodynamic S. fumaroxidans model could not explain why S. fumaroxidans does not produce H2 in monoculture, indicating that current methods might not adequately estimate the thermodynamics, or that other cellular processes (e.g., regulation) play a role. We also developed a thermodynamic coculture model of the association between the organisms. The coculture model correctly predicted the exchange of both H2 and formate between the two species and suggested conditions under which H2 and formate produced by S. fumaroxidans would be fully consumed by M. hungatei. PMID:26147299

  16. Methanogen diversity in the rumen of Indian Surti buffalo (Bubalus bubalis), assessed by 16S rDNA analysis.

    PubMed

    Singh, K M; Tripathi, A K; Pandya, P R; Parnerkar, S; Rank, D N; Kothari, R K; Joshi, C G

    2012-06-01

    The methanogenic communities in buffalo rumen were characterized using a culture-independent approach of a pooled sample of rumen fluid from three adult Surti buffaloes. Buffalo rumen is likely to include species of various methanogens, so 16S rDNA sequences were amplified and cloned from the sample. A total of 171 clones were sequenced to examine 16S rDNA sequence similarity. About 52.63% sequences (90 clones) had ≥ 90% similarity, whereas, 46.78% of the sequences (81 clones) were 75-89% similar to 16S rDNA database sequences, respectively. Phylogenetic analyses were also used to infer the makeup of methanogenic communities in the rumen of Surti buffalo. As a result, we distinguished 23 operational taxonomic units (OTUs) based on unique 16S rDNA sequences: 12 OTUs (52.17%) affiliated to Methanomicrobiales order, 10 OTUs (43.47%) of the order Methanobacteriales and one OTU (4.34%) of Methanosarcina barkeri like clone, respectively. In addition, the population of Methanomicrobiales and Methabacteriales orders were also observed, accounting 4% and 2.17% of total archea. This study has revealed the largest assortment of hydrogenotrophic methanogens phylotypes ever identified from rumen of Surti buffaloes. PMID:21507441

  17. Growth of Methanogens on a Mars Soil Simulant

    NASA Astrophysics Data System (ADS)

    Kral, Timothy A.; Bekkum, Curtis R.; McKay, Christopher P.

    2004-12-01

    Currently, the surface of Mars is probably too cold, too dry, and too oxidizing for life, as we know it, to exist. But the subsurface is another matter. Life forms that might exist below the surface could not obtain their energy from photosynthesis, but rather they would have to utilize chemical energy. Methanogens are one type of microorganism that might be able to survive below the surface of Mars. A potential habitat for existence of methanogens on Mars might be a geothermal source of hydrogen, possibly due to volcanic or hydrothermal activity, or the reaction of basalt and anaerobic water, carbon dioxide, which is abundant in the martian atmosphere, and of course, subsurface liquid water. We report here that certain methanogens can grow on a Mars soil simulant when supplied with carbon dioxide, molecular hydrogen, and varying amounts of water.

  18. Phylogenomic Data Support a Seventh Order of Methylotrophic Methanogens and Provide Insights into the Evolution of Methanogenesis

    PubMed Central

    Borrel, Guillaume; O’Toole, Paul W.; Harris, Hugh M.B.; Peyret, Pierre; Brugère, Jean-François; Gribaldo, Simonetta

    2013-01-01

    Increasing evidence from sequence data from various environments, including the human gut, suggests the existence of a previously unknown putative seventh order of methanogens. The first genomic data from members of this lineage, Methanomassiliicoccus luminyensis and “Candidatus Methanomethylophilus alvus,” provide insights into its evolutionary history and metabolic features. Phylogenetic analysis of ribosomal proteins robustly indicates a monophyletic group independent of any previously known methanogenic order, which shares ancestry with the Marine Benthic Group D, the Marine Group II, the DHVE2 group, and the Thermoplasmatales. This phylogenetic position, along with the analysis of enzymes involved in core methanogenesis, strengthens a single ancient origin of methanogenesis in the Euryarchaeota and indicates further multiple independent losses of this metabolism in nonmethanogenic lineages than previously suggested. Genomic analysis revealed an unprecedented loss of the genes coding for the first six steps of methanogenesis from H2/CO2 and the oxidative part of methylotrophic methanogenesis, consistent with the fact that M. luminyensis and “Ca. M. alvus” are obligate H2-dependent methylotrophic methanogens. Genomic data also suggest that these methanogens may use a large panel of methylated compounds. Phylogenetic analysis including homologs retrieved from environmental samples indicates that methylotrophic methanogenesis (regardless of dependency on H2) is not restricted to gut representatives but may be an ancestral characteristic of the whole order, and possibly also of ancient origin in the Euryarchaeota. 16S rRNA and McrA trees show that this new order of methanogens is very diverse and occupies environments highly relevant for methane production, therefore representing a key lineage to fully understand the diversity and evolution of methanogenesis. PMID:23985970

  19. Bacteroides propionicifaciens sp. nov., isolated from rice-straw residue in a methanogenic reactor treating waste from cattle farms.

    PubMed

    Ueki, Atsuko; Abe, Kunihiro; Kaku, Nobuo; Watanabe, Kazuya; Ueki, Katsuji

    2008-02-01

    Two strictly anaerobic bacterial strains (SV434(T) and S562) were isolated from rice-straw residue in a methanogenic reactor treating waste from cattle farms in Japan. They had identical 16S rRNA gene sequences and showed almost the same phenotypic properties. The cells of both strains were Gram-negative, non-motile, non-spore-forming rods; extraordinarily long rods often occurred. Remarkable stimulation of growth occurred with the addition of haemin and cobalamin (vitamin B(12)) to the medium. The supplementary cobalamin and haemin could be replaced if autoclaved and clarified sludge fluid obtained from the reactor was added. Both strains utilized a range of growth substrates, including arabinose, fructose, galactose, glucose, mannose, cellobiose, maltose, glycogen, starch, dextrin, amygdalin, lactate and pyruvate. Both strains produced acetate and propionate with a small amount of succinate from these substrates in the presence of haemin and cobalamin. Both strains were slightly alkaliphilic, having a pH optimum at about 7.9. The temperature range for growth was 5-35 degrees C, the optimum being 30 degrees C. The NaCl concentration range for growth was 0-4 % (w/v). Catalase activity was not detected in cells cultivated without haemin, whereas cells cultivated with haemin usually had the enzyme activity. Oxidase and nitrate-reducing activities were not detected. Aesculin was hydrolysed, but gelatin was not hydrolysed. Both strains were sensitive to bile acids. The major cellular fatty acids of both strains were anteiso-C(15 : 0) and iso-C(15 : 0). Menaquinones MK-8(H(0)) and MK-9(H(0)) were the major respiratory quinones and the genomic DNA G+C contents were 46.2-47.5 mol%. A phylogenetic analysis based on 16S rRNA gene sequences placed both strains in the phylum Bacteroidetes. Bacteroides coprosuis (isolated from swine-manure storage pits) was the species most closely related to both strains (95.9 % 16S rRNA gene sequence similarity to the type strain). On the

  20. Syntrophic Degradation of Lactate in Methanogenic Co-cultures

    SciTech Connect

    Meyer, Birte; Stahl, David

    2010-05-17

    In environments where the amount of the inorganic electron acceptors (oxygen, nitrate, sulfate, sulfur oroxidized metal ions (Fe3+;Mn4+) is insufficient for complete breakdown of organic matter, methane is formed as the major reduced end product. In such methanogenic environments organic acids are degraded by syntrophic associations of fermenting, acetogenic bacteria (e.g., sulfate-reducing bacteria (SRB) as"secondary fermenters") and methanogenic archaea. In these consortia, the conversion of lactate to acetate, CO2 and methane depends on the cooperating activities of both metabolically distinct microbial groups that are tightly linked by the need to maintain the exchanged metabolites (hydrogenandformate) at very low concentrations.

  1. An ancient divergence among the bacteria. [methanogenic phylogeny

    NASA Technical Reports Server (NTRS)

    Balch, W. E.; Magrum, L. J.; Fox, G. E.; Wolfe, R. S.; Woese, C. R.

    1977-01-01

    The 16S ribosomal RNZs from two species of met methanogenic bacteria, the mesophile Methanobacterium ruminantium and the thermophile Methanobacterium thermoautotrophicum, have been characterized in terms of the oligonucleotides produced by digestion with T1 ribonuclease. These two organisms are found to be sufficiently related that they can be considered members of the same genus or family. However, they bear only slight resemblance to 'typical' Procaryotic genera; such as Escherichia, Bacillus and Anacystis. The divergence of the methanogenic bacteria from other bacteria may be the most ancient phylogenetic event yet detected - antedating considerably the divergence of the blue green algal line for example, from the main bacterial line.

  2. Molecular Signatures of Methanogens in Cultures and Environmental Samples

    NASA Astrophysics Data System (ADS)

    Summons, R. E.; Embaye, T.; Jahnke, L. L.; Baumgartner, M.

    2002-12-01

    The core lipids of methanogens comprise C20 and C40 isoprenoid chains, linked through ether bonds to glycerol. Additional structural diversity is encoded into the polar head groups that are attached to the glycerol ether cores. These compounds are potentially very useful as taxonomic markers in microbial mats and other environmental samples while the nature of the hydrocarbon chains provide a means to identify methanogenic inputs to ancient sediments. The structural diversity of methanogen polar lipids is most valuable when it can be directly correlated to 16S rRNA phylogeny. On the other hand, this diversity can also leads to analytical challenges because there is no single approach that works for all structural types. While some intact methanogen lipids have been identified using mass spectrometry and NMR spectroscopy, the most common means of analysing the lipid cores involves cleavage of the ether bonds using HI and subsequent reduction of the alkyl iodides to hydrocarbons with LiAlH4. One class of methanogenic lipids, the 3?-hydroxyarchaeols, escaped detection for some years because strong acid treatments in the analysis protocols destroyed hydroxyl-containing isoprenoid chains. We have been systematically re-examining the lipids of methanogens, using milder procedures involving weak acid hydrolysis of polar head groups, derivatisation to form trimethylsilyl ethers and analysis by GC-MS. As well as archaeol, sn-2- and sn-3-hydroxyarchaeol, we have tentatively identified a dihydroxyarchaeol in several Methanococcus sp. For Methanococcus thermolithotrophicus an analysis of the total lipid extracts using BBr3 as an ether cleavage reagent followed by LiBEt3H, reduction revealed a very complex mixture consisting of phytane, phytenes, biphytane, biphytenes and a suite of related alcohols. The latter compounds were analysed by GC-MS as their trimethylsilyl ethers and found to comprise a mixture tentatively identified as phytan-N-ol and biphytan-N-ol where N= 3 or 7

  3. Enzymatic lysis of the pseudomurein-containing methanogen Methanobacterium formicicum.

    PubMed Central

    Bush, J W

    1985-01-01

    A streptomycete isolated from cow manure produces an extracellular enzyme capable of lysing the pseudomurein-containing methanogen Methanobacterium formicicum. The lytic activity has been partially purified from culture fluid and appears to be a serine protease. Similar lytic activity has been fractionated from pronase. Optimal conditions have been developed for lysis of M. formicicum by commercial preparations of proteinase K. The three lytic enzymes have been partially characterized. The results with the three enzyme preparations tend to confirm that proteolytic enzymes are capable of lysing methanogen cells. Images PMID:3891731

  4. Snapshot of methanogen sensitivity to temperature in Zoige wetland from Tibetan plateau

    PubMed Central

    Fu, Li; Song, Tianze; Lu, Yahai

    2015-01-01

    Zoige wetland in Tibetan plateau represents a cold environment at high altitude where significant methane emission has been observed. However, it remains unknown how the production and emission of CH4 from Zoige wetland will respond to a warming climate. Here we investigated the temperature sensitivity of methanogen community in a Zoige wetland soil under the laboratory incubation conditions. One soil sample was collected and the temperature sensitivity of the methanogenic activity, the structure of methanogen community and the methanogenic pathways were determined. We found that the response of methanogenesis to temperature could be separated into two phases, a high sensitivity in the low temperature range and a modest sensitivity under mesophilic conditions, respectively. The aceticlastic methanogens Methanosarcinaceae were the main methanogens at low temperatures, while hydrogenotrophic Methanobacteriales, Methanomicrobiales, and Methanocellales were more abundant at higher temperatures. The total abundance of mcrA genes increased with temperature indicating that the growth of methanogens was stimulated. The growth of hydrogenotrophic methanogens, however, was faster than aceticlastic ones resulting in the shift of methanogen community. Determination of carbon isotopic signatures indicated that methanogenic pathway was also shifted from mainly aceticlastic methanogenesis to a mixture of hydrogenotrophic and aceticlastic methanogenesis with the increase of temperature. Collectively, the shift of temperature responses of methanogenesis was in accordance with the changes in methanogen composition and methanogenic pathway in this wetland sample. It appears that the aceticlastic methanogenesis dominating at low temperatures is more sensitive than the hydrogenotrophic one at higher temperatures. PMID:25745422

  5. Effects of Spartina alterniflora invasion on the communities of methanogens and sulfate-reducing bacteria in estuarine marsh sediments

    PubMed Central

    Zeleke, Jemaneh; Sheng, Qiang; Wang, Jian-Gong; Huang, Ming-Yao; Xia, Fei; Wu, Ji-Hua; Quan, Zhe-Xue

    2013-01-01

    The effect of plant invasion on the microorganisms of soil sediments is very important for estuary ecology. The community structures of methanogens and sulfate-reducing bacteria (SRB) as a function of Spartina alterniflora invasion in Phragmites australis-vegetated sediments of the Dongtan wetland in the Yangtze River estuary, China, were investigated using 454 pyrosequencing and quantitative real-time PCR (qPCR) of the methyl coenzyme M reductase A (mcrA) and dissimilatory sulfite-reductase (dsrB) genes. Sediment samples were collected from two replicate locations, and each location included three sampling stands each covered by monocultures of P. australis, S. alterniflora and both plants (transition stands), respectively. qPCR analysis revealed higher copy numbers of mcrA genes in sediments from S. alterniflora stands than P. australis stands (5- and 7.5-fold more in the spring and summer, respectively), which is consistent with the higher methane flux rates measured in the S. alterniflora stands (up to 8.01 ± 5.61 mg m−2 h−1). Similar trends were observed for SRB, and they were up to two orders of magnitude higher than the methanogens. Diversity indices indicated a lower diversity of methanogens in the S. alterniflora stands than the P. australis stands. In contrast, insignificant variations were observed in the diversity of SRB with the invasion. Although Methanomicrobiales and Methanococcales, the hydrogenotrophic methanogens, dominated in the salt marsh, Methanomicrobiales displayed a slight increase with the invasion and growth of S. alterniflora, whereas the later responded differently. Methanosarcina, the metabolically diverse methanogens, did not vary with the invasion of, but Methanosaeta, the exclusive acetate utilizers, appeared to increase with S. alterniflora invasion. In SRB, sequences closely related to the families Desulfobacteraceae and Desulfobulbaceae dominated in the salt marsh, although they displayed minimal changes with the S

  6. Effective Suppression of Methane Emission by 2-Bromoethanesulfonate during Rice Cultivation

    PubMed Central

    Waghmode, Tatoba R.; Haque, Md. Mozammel; Kim, Sang Yoon; Kim, Pil Joo

    2015-01-01

    2-bromoethanesulfonate (BES) is a structural analogue of coenzyme M (Co-M) and potent inhibitor of methanogenesis. Several studies confirmed, BES can inhibit CH4 prodcution in rice soil, but the suppressing effectiveness of BES application on CH4 emission under rice cultivation has not been studied. In this pot experiment, different levels of BES (0, 20, 40 and 80 mg kg-1) were applied to study its effect on CH4 emission and plant growth during rice cultivation. Application of BES effectively suppressed CH4 emission when compared with control soil during rice cultivation. The CH4 emission rates were significantly (P<0.001) decreased by BES application possibly due to significant (P<0.001) reduction of methnaogenic biomarkers like Co-M concentration and mcrA gene copy number (i.e. methanogenic abunadance). BES significantly (P<0.001) reduced methanogen activity, while it did not affect soil dehydrogenase activity during rice cultivation. A rice plant growth and yield parameters were not affected by BES application. The maximum CH4 reduction (49% reduction over control) was found at 80 mg kg-1 BES application during rice cultivation. It is, therefore, concluded that BES could be a suitable soil amendment for reducing CH4 emission without affecting rice plant growth and productivity during rice cultivation. PMID:26562416

  7. Effective Suppression of Methane Emission by 2-Bromoethanesulfonate during Rice Cultivation.

    PubMed

    Waghmode, Tatoba R; Haque, Md Mozammel; Kim, Sang Yoon; Kim, Pil Joo

    2015-01-01

    2-bromoethanesulfonate (BES) is a structural analogue of coenzyme M (Co-M) and potent inhibitor of methanogenesis. Several studies confirmed, BES can inhibit CH4 prodcution in rice soil, but the suppressing effectiveness of BES application on CH4 emission under rice cultivation has not been studied. In this pot experiment, different levels of BES (0, 20, 40 and 80 mg kg-1) were applied to study its effect on CH4 emission and plant growth during rice cultivation. Application of BES effectively suppressed CH4 emission when compared with control soil during rice cultivation. The CH4 emission rates were significantly (P<0.001) decreased by BES application possibly due to significant (P<0.001) reduction of methnaogenic biomarkers like Co-M concentration and mcrA gene copy number (i.e. methanogenic abunadance). BES significantly (P<0.001) reduced methanogen activity, while it did not affect soil dehydrogenase activity during rice cultivation. A rice plant growth and yield parameters were not affected by BES application. The maximum CH4 reduction (49% reduction over control) was found at 80 mg kg-1 BES application during rice cultivation. It is, therefore, concluded that BES could be a suitable soil amendment for reducing CH4 emission without affecting rice plant growth and productivity during rice cultivation. PMID:26562416

  8. NATURAL ATTENUATION OF MTBE IN THE SUBSURFACE UNDER METHANOGENIC CONDITIONS

    EPA Science Inventory

    This case study was conducted at the former Fuel Farm Site at the U.S.Coast Guard Support Center at Elizabeth City, North Carolina. The study is intended to answer the following questions. Can MTBE be biodegraded under methanogenic conditions in ground water that was contaminated...

  9. Methane as a product of chloroethene biodegradation under methanogenic conditions

    USGS Publications Warehouse

    Bradley, P.M.; Chapelle, F.H.

    1999-01-01

    Radiometric detection headspace analyses of microcosms containing bed sediments from two geographically distinct sites indicated that 10-39% of the radiolabeled carbon transformed during anaerobic biodegradation of [1,2- 14C]trichloroethene (TCE) or [1,2-14C]vinyl chloride (VC) under methanogenic conditions was ultimately incorporated into 14CH4. The results demonstrate that, in addition to ethene, ethane, and CO2, CH4 can be a significant product of chloroethene biodegradation in some methanogenic sediments.Radiometric detection headspace analyses of microcosms containing bed sediments from two geographically distinct sites indicated that 10-39% of the radiolabeled carbon transformed during anaerobic biodegradation of [1,2-14C]trichloroethene (TCE) or [1,2-14C]vinyl chloride (VC) under methanogenic conditions was ultimately incorporated into 14CH4. The results demonstrate that, in addition to ethene, ethane, and CO2, CH4 can be a significant product of chloroethene biodegradation in some methanogenic sediments.

  10. Survival of methanogens during desiccation: implications for life on Mars.

    PubMed

    Kendrick, Michael G; Kral, Timothy A

    2006-08-01

    The relatively recent discoveries that liquid water likely existed on the surface of past Mars and that methane currently exists in the martian atmosphere have fueled the possibility of extant or extinct life on Mars. One possible explanation for the existence of the methane would be the presence of methanogens in the subsurface. Methanogens are microorganisms in the domain Archaea that can metabolize molecular hydrogen as an energy source and carbon dioxide as a carbon source and produce methane. One factor of importance is the arid nature of Mars, at least at the surface. If one is to assume that life exists below the surface, then based on the only example of life that we know, liquid water must be present. Realistically, however, that liquid water may be seasonal just as it is at some locations on our home planet. Here we report on research designed to determine how long certain species of methanogens can survive desiccation on a Mars soil simulant, JSC Mars-1. Methanogenic cells were grown on JSC Mars-1, transferred to a desiccator within a Coy anaerobic environmental chamber, and maintained there for varying time periods. Following removal from the desiccator and rehydration, gas chromatographic measurements of methane indicated survival for varying time periods. Methanosarcina barkeri survived desiccation for 10 days, while Methanobacterium formicicum and Methanothermobacter wolfeii were able to survive for 25 days. PMID:16916281

  11. Survival of Methanogens During Desiccation: Implications for Life on Mars

    NASA Astrophysics Data System (ADS)

    Kendrick, Michael G.; Kral, Timothy A.

    2006-08-01

    The relatively recent discoveries that liquid water likely existed on the surface of past Mars and that methane currently exists in the martian atmosphere have fueled the possibility of extant or extinct life on Mars. One possible explanation for the existence of the methane would be the presence of methanogens in the subsurface. Methanogens are microorganisms in the domain Archaea that can metabolize molecular hydrogen as an energy source and carbon dioxide as a carbon source and produce methane. One factor of importance is the arid nature of Mars, at least at the surface. If one is to assume that life exists below the surface, then based on the only example of life that we know, liquid water must be present. Realistically, however, that liquid water may be seasonal just as it is at some locations on our home planet. Here we report on research designed to determine how long certain species of methanogens can survive desiccation on a Mars soil simulant, JSC Mars-1. Methanogenic cells were grown on JSC Mars-1, transferred to a desiccator within a Coy anaerobic environmental chamber, and maintained there for varying time periods. Following removal from the desiccator and rehydration, gas chromatographic measurements of methane indicated survival for varying time periods. Methanosarcina barkeri survived desiccation for 10 days, while Methanobacterium formicicum and Methanothermobacter wolfeii were able to survive for 25 days.

  12. Metabolism of 3-methylindole by a methanogenic consortium

    SciTech Connect

    Jidong Gu; Berry, D.F. )

    1992-08-01

    A methanogenic 3-methylindole (3-MI)-degrading consortium, enriched from wetland soil, completely mineralized 3-MI. Degradation proceeded through an initial hydroxylation reaction forming 3-methyloxindole. The consortium was unable to degrade oxindole or isatin, suggesting a new pathway for 3-MI fermentation. 3-Methylindole was identified by mass spectroscopy, ultraviolet spectroscopy, and proton nuclear magnetic resonance spectrometry.

  13. Multiple Syntrophic Interactions in a Terephthalate-Degrading Methanogenic Consortium

    SciTech Connect

    Lykidis, Athanasios; Chen, Chia-Lung; Tringe, Susannah G.; McHardy, Alice C.; Copeland, Alex 5; Kyrpides, Nikos C.; Hugenholtz, Philip; Liu, Wen-Tso

    2010-08-05

    Terephthalate (TA) is one of the top 50 chemicals produced worldwide. Its production results in a TA-containing wastewater that is treated by anaerobic processes through a poorly understood methanogenic syntrophy. Using metagenomics, we characterized the methanogenic consortium tinside a hyper-mesophilic (i.e., between mesophilic and thermophilic), TA-degrading bioreactor. We identified genes belonging to dominant Pelotomaculum species presumably involved in TA degradation through decarboxylation, dearomatization, and modified ?-oxidation to H{sub 2}/CO{sub 2} and acetate. These intermediates are converted to CH{sub 4}/CO{sub 2} by three novel hyper-mesophilic methanogens. Additional secondary syntrophic interactions were predicted in Thermotogae, Syntrophus and candidate phyla OP5 and WWE1 populations. The OP5 encodes genes capable of anaerobic autotrophic butyrate production and Thermotogae, Syntrophus and WWE1 have the genetic potential to oxidize butyrate to COsub 2}/H{sub 2} and acetate. These observations suggest that the TA-degrading consortium consists of additional syntrophic interactions beyond the standard H{sub 2}-producing syntroph ? methanogen partnership that may serve to improve community stability.

  14. Methanogenic community shift in anaerobic batch digesters treating swine wastewater.

    PubMed

    Kim, Woong; Lee, Seungyong; Shin, Seung Gu; Lee, Changsoo; Hwang, Kwanghyun; Hwang, Seokhwan

    2010-09-01

    Qualitative and quantitative molecular analysis techniques were used to determine associations between differences in methanogenic microbial communities and the efficiency of batch anaerobic digesters. Two bioreactors were initially seeded with anaerobic sludge originating from a local municipal wastewater treatment plant and then supplemented with swine wastewater. Differences were observed in the total amount of methane produced in the two bioreactors (7.9L/L, and 4.5L/L, respectively). To explain these differences, efforts were taken to characterize the microbial populations present using a PCR-based DGGE analysis with methanogenic primer and probe sets. The groups Methanomicrobiales (MMB), Methanobacteriales (MBT), and Methanosarcinales (MSL) were detected, but Methanococcales (MCC) was not detected. Following this qualitative assay, real-time PCR was used to investigate quantitative differences in the populations of these methanogenic orders. MMB was found to be the dominant order present and its abundance patterns were different in the two digesters. The population profiles of the other methanogenic groups also differed. Through redundancy analysis, correlations between the concentrations of the different microbes and chemical properties such as volatile fatty acids were calculated. Correlations between MBT and MSL populations and chemical properties were found to be consistent in both digesters, however, differences were observed in the correlations between MMB and propionate. These results suggest that interactions between populations of MMB and other methanogens affected the final methane yield, despite MMB remaining the dominant group overall. The exact details of why changes in the MMB community caused different profiles of methane production could not be ascertained. However, this research provides evidence that microbial behavior is important for regulating the performance of anaerobic processes. PMID:20692007

  15. Anaerobic degradation of phthalate isomers by methanogenic consortia

    SciTech Connect

    Kleerebezem, R.; Pol, L.W.H.; Lettinga, G.

    1999-03-01

    Three methanogenic enrichment cultures, grown on ortho-phthalate, iso-phthalate, or terephthalate were obtained from digested sewage sludge or methanogenic granular sludge. Cultures grown on one of the phthalate isomers were not capable of degrading the other phthalate isomers. All three cultures had the ability to degrade benzoate. Maximum specific growth rates ({mu}{sub S}{sup max}) and biomass yields (Y{sub X{sub tot}S}) of the mixed cultures were determined by using both the phthalate isomers and benzoate as substrates. Comparable values for these parameters were found for all three cultures. Values for {mu}{sub X}{sup max} and Y{sub X{sub tot}S} were higher for growth on benzoate compared to the phthalate isomers. Based on measured and estimated values for the microbial yield of the methanogens in the mixed culture, specific yields for the phthalate and benzoate fermenting organisms were calculated. A kinetic model, involving three microbial species, was developed to predict intermediate acetate and hydrogen accumulation and the final production of methane. Values for the ratio of the concentrations of methanogenic organisms, versus the phthalate isomer and benzoate fermenting organisms, and apparent half-saturation constants (K{sub S}) for the methanogens were calculated. By using this combination of measured and estimated parameter values, a reasonable description of intermediate accumulation and methane formation was obtained, with the initial concentration of phthalate fermenting organisms being the only variable. The energetic efficiency for growth of the fermenting organisms on the phthalate isomers was calculated to be significantly smaller than for growth on benzoate.

  16. Ruminal Methanogen Community in Dairy Cows Fed Agricultural Residues of Corn Stover, Rapeseed, and Cottonseed Meals.

    PubMed

    Wang, Pengpeng; Zhao, Shengguo; Wang, Xingwen; Zhang, Yangdong; Zheng, Nan; Wang, Jiaqi

    2016-07-13

    The purpose was to reveal changes in the methanogen community in the rumen of dairy cows fed agricultural residues of corn stover, rapeseed, and cottonseed meals, compared with alfalfa hay or soybean meal. Analysis was based on cloning and sequencing the methyl coenzyme M reductase α-subunit gene of ruminal methanogens. Results revealed that predicted methane production was increased while population of ruminal methanogens was not significantly affected when cows were fed diets containing various amounts of agricultural residues. Richness and diversity of methanogen community were markedly increased by addition of agricultural residues. The dominant ruminal methanogens shared by all experimental groups belonged to rumen cluster C, accounting for 71% of total, followed by the order Methanobacteriales (29%). Alterations of ruminal methanogen community and prevalence of particular species occurred in response to fed agricultural residue rations, suggesting the possibility of regulating target methanogens to control methane production by dairy cows fed agricultural residues. PMID:27322573

  17. Linkage among Vegetation, Microbes and Methanogenic Pathways in Alaskan Peatlands

    NASA Astrophysics Data System (ADS)

    Zhang, L.; Sidelinger, W.; Shu, H.; Varner, R. K.; Hines, M. E.

    2014-12-01

    Northern wetlands are thought to account for one third of the naturally emitted CH4. However, methane production pathways in northern peatlands are poorly understood, yet are predicted to change in response to vegetation shifts due to warming. Previous studies noted that acetate conversion to methane (acetoclastic methanogenesis, AM) in northern wetlands is largely impeded and acetate accumulates, however AM tends to increase with minerotrophy. To understand methanogenic pathways and to provide linkage among pathways, we studied Alaskan wetlands in 2013 and 2014. In 2013, laboratory incubations were conducted in three peatlands representing trophic gradients from bogs to fens. During 2014, 37 different sites in Fairbanks and Anchorage were studied that represented wetlands with pH values from 3.5 to 5.5 and vegetation from primarily Sphagnum to sedges. Measurements in 2014 included vegetation composition, gases (CH4, CO2, H2, and CO), 13CH4 and 13CO2, volatile fatty acids, DOC, other electron acceptors. Further incubation studies are being conducted to decipher controls on decomposition pathways. Gene sequencing was used to characterize microbial community composition, and metagenomic and transcriptomics were conducted to describe community activity. Results showed that methanogenesis was higher in fens than bogs, but hydrogenotrophic methanogenesis (HM) was dominant at all sites. End product ratios showed that AM was occurring in fens, albeit slowly. Fermentation was an important end-point in decomposition and microbial syntrophy was weak. These data, regardless of trophic status, differed greatly from data obtained from temperate wetlands in which terminal respiratory processes were strong and C flow through syntrophy was important. Trophic status influenced C flow in the Alaskan sites, but terminal processes were weak and end product formation tended to end at primary fermentation, which dominated as the terminal step in decomposition.

  18. Complete genome sequence of Methanolinea tarda NOBI-1T, a hydrogenotrophic methanogen isolated from methanogenic digester sludge

    DOE PAGESBeta

    Yamamoto, Kyosuke; Tamaki, Hideyuki; Cadillo-Quiroz, Hinsby; Imachi, Hiroyuki; Kyrpides, Nikos; Woyke, Tanja; Goodwin, Lynne; Zinder, Stephen H.; Kamagata, Yoichi; Liu, Wen -Tso

    2014-09-04

    In this study, we report a 2.0-Mb complete genome sequence of Methanolinea tarda NOBI-1T, a methanogenic archaeon isolated from an anaerobic digested sludge. This is the first genome report of the genus Methanolinea isolate belonging to the family Methanoregulaceae, a recently proposed novel family within the order Methanomicrobiales.

  19. How Specific Microbial Communities Benefit the Oil Industry: Significant Contribution of Methyl/Methanol-Utilising Methanogenic Pathway in a Subsurface Biogas Environment

    NASA Astrophysics Data System (ADS)

    Strąpoć, Dariusz; Ashby, Matt; Wood, Ladonna; Levinson, Rick; Huizinga, Bradley

    Methanogenesis is considered the main terminal process of subsurface anaerobic organic-matter degradation. Previous geochemical studies have reported CO2-reducing and acetoclastic methanogenesis as the predominant subsurface methanogenic pathways for primary and secondary biogenic gas generation (i.e. in oil biodegradation or coalbed methane settings). In lab-scale experiments and microbiology literature, however, methanogens have been shown to be able to utilise a wider variety of substrates, typically containing methyl groups, i.e. dimethyl sulphide (DMS), methyl amines (e.g. TMA), formate, and methanol. Additional methanogenic substrates include CO and other primary alcohols and secondary alcohols (Whitman et al., 2006; Fig. 25.1). Here, we describe a volumetrically important natural biogenic gas field in which these methylotrophic pathways have contributed significantly to biomethane formation.

  20. The alkaloid gramine in the anaerobic digestion process-inhibition and adaptation of the methanogenic community.

    PubMed

    Popp, Denny; Harms, Hauke; Sträuber, Heike

    2016-08-01

    As many plant secondary metabolites have antimicrobial activity, microorganisms of the anaerobic digestion process might be affected when plant material rich in these compounds is digested. Hitherto, the effects of plant secondary metabolites on the anaerobic digestion process are poorly investigated. In this study, the alkaloid gramine, a constituent of reed canary grass, was added daily to a continuous co-digestion of grass silage and cow manure. A transient decrease of the methane yield by 17 % and a subsequent recovery was observed, but no effect on other process parameters. When gramine was infrequently spiked in higher amounts, the observed inhibitory effect was even more pronounced including a 53 % decrease of the methane yield and an increase of acetic acid concentrations up to 96 mM. However, the process recovered and the process parameters were finally at initial values (methane yield around 255 LN CH4 per gram volatile solids of substrate and acetic acid concentration lower than 2 mM). The bacterial communities of the reactors remained stable upon gramine addition. In contrast, the methanogenic community changed from a well-balanced mixture of five phylotypes towards a strong dominance of Methanosarcina (more than two thirds of the methanogenic community) while Methanosaeta disappeared. Batch inhibition assays revealed that acetic acid was only converted to methane via acetoclastic methanogenesis which was more strongly affected by gramine than hydrogenotrophic methanogenesis and acetogenesis. Hence, when acetoclastic methanogenesis is the dominant pathway, a shift of the methanogenic community is necessary to digest gramine-rich plant material. PMID:27138201

  1. Flaxseed supplementation decreases methanogenic gene abundance in the rumen of dairy cows.

    PubMed

    Li, L; Schoenhals, K E; Brady, P A; Estill, C T; Perumbakkam, S; Craig, A M

    2012-11-01

    The objective of this study was to investigate the effects of a flaxseed-supplemented diet on archaeal abundance and gene expression of methanogens in the rumen of dairy cows. In all, 11 non-lactating dairy cows were randomly divided into two groups: group A (five cows) and B (six cows). The two diets fed were: (1) the control diet, a conventional dry cow ration; and (2) the flaxseed-supplemented diet, the conventional dry cow ration adjusted with 12.16% ground flaxseed incorporated into the total mixed ration. A cross-over experiment was performed with the two groups of cows fed the two different diets for five 21-day periods, which included the first adaptation period followed by two treatment and two wash out periods. At the end of each feeding period, rumen fluid samples were collected via rumenocentesis and DNA was extracted. Quantitative PCR was utilized to analyze the gene abundance of 16S ribosomal RNA (16S rRNA) targeting the ruminal archaea population and the mcrA gene coding for methyl coenzyme-M reductase subunit A, a terminal enzyme in the methanogenesis pathway. Results demonstrated a 49% reduction of 16S rRNA and 50% reduction of mcrA gene abundances in the rumen of dairy cows fed the flaxseed-supplemented diet in comparison with those fed the control diet. This shows flaxseed supplementation effectively decreases the methanogenic population in the rumen. Future studies will focus on the mechanisms for such reduction in the rumen of dairy cattle, as well as the relationship between methanogenic gene expression and methane production. PMID:22717375

  2. Cultivating Mentors.

    ERIC Educational Resources Information Center

    McGee, Rick; Cody, Heather Heim

    1995-01-01

    Describes the Mayo Minority Scholars Program, which helps American Indian and other minority students become doctors, medical faculty, or biomedical researchers by matching them with appropriate mentors. Relates the authors' perspectives and experiences as mentor and student. Includes program and contact information. (JAT)

  3. Finding a robust strain for biomethanation: anaerobic fungi (Neocallimastigomycota) from the Alpine ibex (Capra ibex) and their associated methanogens.

    PubMed

    Leis, Stefanie; Dresch, Philipp; Peintner, Ursula; Fliegerová, Katerina; Sandbichler, Adolf Michael; Insam, Heribert; Podmirseg, Sabine Marie

    2014-10-01

    Anaerobic fungi occupy the rumen and digestive tract of herbivores, where they play an important role in enzymatic digestion of lignocellulosic and cellulosic substrates, i.e. organic material that their hosts are unable to decompose on their own. In this study we isolated anaerobic fungi from a typical alpine herbivore, the Alpine ibex (C. ibex). Three fungal strains, either as pure culture (ST2) or syntrophic co-culture with methanogens (ST3, ST4) were successfully obtained and morphologically characterised by different microscopy- and staining-techniques and by rDNA ITS gene sequencing. The isolated fungi were identified as Neocallimastix frontalis (ST2) and Caecomyces communis (ST3 and ST4). We introduce a novel field of application for lactofuchsin-staining, combined with confocal laser scanning microscopy. This approach proved as an effective method to visualize fungal structures, especially in the presence of plant biomass, generally exhibiting high autofluorescence. Moreover, we could demonstrate that fungal morphology is subject to changes depending on the carbon source used for cultivation. Oxygen tolerance was confirmed for both, C. communis-cultures for up to three, and for the N. frontalis-isolate for up to 12 h, respectively. With PCR, FISH and an oligonucleotide microarray we found associated methanogens (mainly Methanobacteriales) for C. communis, but not for N. frontalis. PMID:24384307

  4. Methanogen prevalence throughout the gastrointestinal tract of pre-weaned dairy calves

    PubMed Central

    Zhou, Mi; Chen, Yanhong; Griebel, Philip J; Guan, Le Luo

    2014-01-01

    The methanogenic community throughout the gastrointestinal tract (GIT) of pre-weaned calves has not been well studied. The current study firstly investigated the distribution and composition of the methanogenic community in the rumen, ileum, and colon of 3–4 week-old milk-fed dairy calves (n = 4) using 16S rRNA gene clone library analysis. The occurrence of methanogens in the GIT of pre-weaned calves was further validated by using PCR-denaturing gradient gel electrophoresis (PCR-DGGE), and quantitative real-time PCR (qPCR) was applied to quantify the methanogenic community in the rumen, jejunum, ileum, cecum, colon and rectum of 8 3–4 week old animals. Both cloning libraries and PCR-DGGE revealed that phylotypes close to Methanobrevibacter were the main taxon along the GIT in pre-weaned sucking calves. The composition and abundance of methanogens varied significantly among individual animals, suggesting that host conditions may influence the composition of the symbiotic microbiota. Segregation of methanogenic communities throughout the GIT was also observed within individual animals, suggesting possible functional differences among methanogens residing in different GIT regions. This is the first study to analyze methanogenic communities throughout the GIT of milk-fed newborn dairy calves and reveal both their diversity and abundance. The identification of methanogens in the lower GIT of pre-weaned dairy calves warrants further investigation to better define methanogen roles in GIT function and their impact on host metabolism and health. PMID:25483332

  5. The complete genome sequence of the rumen methanogen Methanobrevibacter millerae SM9.

    PubMed

    Kelly, William J; Pacheco, Diana M; Li, Dong; Attwood, Graeme T; Altermann, Eric; Leahy, Sinead C

    2016-01-01

    Methanobrevibacter millerae SM9 was isolated from the rumen of a sheep maintained on a fresh forage diet, and its genome has been sequenced to provide information on the phylogenetic diversity of rumen methanogens with a view to developing technologies for methane mitigation. It is the first rumen isolate from the Methanobrevibacter gottschalkii clade to have its genome sequence completed. The 2.54 Mb SM9 chromosome has an average G + C content of 31.8 %, encodes 2269 protein-coding genes, and harbors a single prophage. The overall gene content is comparable to that of Methanobrevibacter ruminantium M1 and the type strain of M. millerae (ZA-10(T)) suggesting that the basic metabolism of these two hydrogenotrophic rumen methanogen species is similar. However, M. millerae has a larger complement of genes involved in methanogenesis including genes for methyl coenzyme M reductase II (mrtAGDB) which are not found in M1. Unusual features of the M. millerae genomes include the presence of a tannase gene which shows high sequence similarity with the tannase from Lactobacillus plantarum, and large non-ribosomal peptide synthase genes. The M. millerae sequences indicate that methane mitigation strategies based on the M. ruminantium M1 genome sequence are also likely to be applicable to members of the M. gottschalkii clade. PMID:27536339

  6. Vaccination of Sheep with a Methanogen Protein Provides Insight into Levels of Antibody in Saliva Needed to Target Ruminal Methanogens

    PubMed Central

    Subharat, Supatsak; Shu, Dairu; Zheng, Tao; Buddle, Bryce M.; Kaneko, Kan; Hook, Sarah; Janssen, Peter H.; Wedlock, D. Neil

    2016-01-01

    Methane is produced in the rumen of ruminant livestock by methanogens and is a major contributor to agricultural greenhouse gases. Vaccination against ruminal methanogens could reduce methane emissions by inducing antibodies in saliva which enter the rumen and impair ability of methanogens to produce methane. Presently, it is not known if vaccination can induce sufficient amounts of antibody in the saliva to target methanogen populations in the rumen and little is known about how long antibody in the rumen remains active. In the current study, sheep were vaccinated twice at a 3-week interval with a model methanogen antigen, recombinant glycosyl transferase protein (rGT2) formulated with one of four adjuvants: saponin, Montanide ISA61, a chitosan thermogel, or a lipid nanoparticle/cationic liposome adjuvant (n = 6/formulation). A control group of sheep (n = 6) was not vaccinated. The highest antigen-specific IgA and IgG responses in both saliva and serum were observed with Montanide ISA61, which promoted levels of salivary antibodies that were five-fold higher than the second most potent adjuvant, saponin. A rGT2-specific IgG standard was used to determine the level of rGT2-specific IgG in serum and saliva. Vaccination with GT2/Montanide ISA61 produced a peak antibody concentration of 7 × 1016 molecules of antigen-specific IgG per litre of saliva, and it was estimated that in the rumen there would be more than 104 molecules of antigen-specific IgG for each methanogen cell. Both IgG and IgA in saliva were shown to be relatively stable in the rumen. Salivary antibody exposed for 1–2 hours to an in vitro simulated rumen environment retained approximately 50% of antigen-binding activity. Collectively, the results from measuring antibody levels and stablility suggest a vaccination-based mitigation strategy for livestock generated methane is in theory feasible. PMID:27472482

  7. Vaccination of Sheep with a Methanogen Protein Provides Insight into Levels of Antibody in Saliva Needed to Target Ruminal Methanogens.

    PubMed

    Subharat, Supatsak; Shu, Dairu; Zheng, Tao; Buddle, Bryce M; Kaneko, Kan; Hook, Sarah; Janssen, Peter H; Wedlock, D Neil

    2016-01-01

    Methane is produced in the rumen of ruminant livestock by methanogens and is a major contributor to agricultural greenhouse gases. Vaccination against ruminal methanogens could reduce methane emissions by inducing antibodies in saliva which enter the rumen and impair ability of methanogens to produce methane. Presently, it is not known if vaccination can induce sufficient amounts of antibody in the saliva to target methanogen populations in the rumen and little is known about how long antibody in the rumen remains active. In the current study, sheep were vaccinated twice at a 3-week interval with a model methanogen antigen, recombinant glycosyl transferase protein (rGT2) formulated with one of four adjuvants: saponin, Montanide ISA61, a chitosan thermogel, or a lipid nanoparticle/cationic liposome adjuvant (n = 6/formulation). A control group of sheep (n = 6) was not vaccinated. The highest antigen-specific IgA and IgG responses in both saliva and serum were observed with Montanide ISA61, which promoted levels of salivary antibodies that were five-fold higher than the second most potent adjuvant, saponin. A rGT2-specific IgG standard was used to determine the level of rGT2-specific IgG in serum and saliva. Vaccination with GT2/Montanide ISA61 produced a peak antibody concentration of 7 × 1016 molecules of antigen-specific IgG per litre of saliva, and it was estimated that in the rumen there would be more than 104 molecules of antigen-specific IgG for each methanogen cell. Both IgG and IgA in saliva were shown to be relatively stable in the rumen. Salivary antibody exposed for 1-2 hours to an in vitro simulated rumen environment retained approximately 50% of antigen-binding activity. Collectively, the results from measuring antibody levels and stablility suggest a vaccination-based mitigation strategy for livestock generated methane is in theory feasible. PMID:27472482

  8. Methanohalophilus zhilinae sp. nov., an alkaliphilic, halophilic, methylotrophic methanogen

    NASA Technical Reports Server (NTRS)

    Mathrani, I. M.; Boone, D. R.; Mah, R. A.; Fox, G. E.; Lau, P. P.

    1988-01-01

    Methanohalophilus zhilinae, a new alkaliphilic, halophilic, methylotrophic species of methanogenic bacteria, is described. Strain WeN5T (T = type strain) from Bosa Lake of the Wadi el Natrun in Egypt was designated the type strain and was further characterized. This strain was nonmotile, able to catabolize dimethylsulfide, and able to grow in medium with a methyl group-containing substrate (such as methanol or trimethylamine) as the sole organic compound added. Sulfide (21 mM) inhibited cultures growing on trimethylamine. The antibiotic susceptibility pattern of strain WeN5T was typical of the pattern for archaeobacteria, and the guanine-plus-cytosine content of the deoxyribonucleic acid was 38 mol%. Characterization of the 16S ribosomal ribonucleic acid sequence indicated that strain WeN5T is phylogenetically distinct from members of previously described genera other than Methanohalophilus and supported the partition of halophilic methanogens into their own genus.

  9. Composition and Role of Extracellular Polymers in Methanogenic Granules

    PubMed Central

    Veiga, M. C.; Jain, M. K.; Wu, W.; Hollingsworth, R. I.; Zeikus, J. G.

    1997-01-01

    Methanobacterium formicicum and Methanosarcina mazeii are two prevalent species isolated from an anaerobic granular consortium grown on a fatty acid mixture. The extracellular polysaccharides (EPS) were extracted from Methanobacterium formicicum and Methanosarcina mazeii and from the methanogenic granules to examine their role in granular development. The EPS made up approximately 20 to 14% of the extracellular polymer extracted from the granules, Methanobacterium formicicum, and Methanosarcina mazeii. The EPS produced by Methanobacterium formicicum was composed mainly of rhamnose, mannose, galactose, glucose, and amino sugars, while that produced by Methanosarcina mazeii contained ribose, galactose, glucose, and glucosamine. The same sugars were also present in the EPS produced by the granules. These results indicate that the two methanogens, especially Methanobacterium formicicum, contributed significantly to the production of the extracellular polymer of the anaerobic granules. Growth temperature, substrates (formate and H(inf2)-CO(inf2)), and the key nutrients (nitrogen and phosphate concentrations) affected polymer production by Methanobacterium formicicum. PMID:16535504

  10. Geographic information system-based identification of suitable cultivation sites for wood-cultivated ginseng.

    PubMed

    Beon, Mu Sup; Park, Jun Ho; Kang, Hag Mo; Cho, Sung Jong; Kim, Hyun

    2013-10-01

    Wood-cultivated ginseng, including roots in its dried form, is produced in forest land without using artificial facilities such as light barriers. To identify suitable sites for the propagation of wood-cultivated ginseng, factor combination technique (FCT) and linear combination technique (LCT) were used with geographic information system and the results were superimposed onto an actual wood-cultivated ginseng plantation. The LCT more extensively searched for suitable sites of cultivation than that by the FCT; further, the LCT probed wide areas considering the predominance of precipitous mountains in Korea. In addition, the LCT showed the much higher degree of overlap with the actual cultivation sites; therefore, the LCT more comprehensively reflects the cultivator's intention for site selection. On the other hand, the inclusion of additional factors for the selection of suitable cultivation sites and experts' opinions may enhance the effectiveness and accuracy of the LCT for site application. PMID:24235864

  11. Reducing methane emissions in sheep by immunization against rumen methanogens.

    PubMed

    Wright, A D G; Kennedy, P; O'Neill, C J; Toovey, A F; Popovski, S; Rea, S M; Pimm, C L; Klein, L

    2004-09-28

    This work was conducted to determine if methane emissions from sheep immunized with an anti-methanogen vaccine were significantly lower than methane emissions from non-immunized sheep, to test the effectiveness of two different vaccine formulations (VF) on methane abatement, and to compare methane emissions measured using a closed-circuit respiration chamber and the sulphur-hexafluoride (SF6) tracer technique. Thirty mature wether sheep were randomly allocated to three treatment groups (n = 10). One group received an immunization of adjuvant only on days 0 and 153 (control), a second group received an immunization with a 3-methanogen mix on days 0 and 153 (VF3 + 3), and a third group received an immunization of a 7-methanogen mix on day 0 followed by a 3-methanogen mix on day 153 (VF7 + 3). Four weeks post-secondary immunization, there was a significant 7.7% reduction in methane production per kg dry matter intake in the VF7 + 3 group compared to the controls (P = 0.051). However, methane emissions from sheep immunized with VF7 + 3 were not significantly different when compared to the sheep in the control group (P = 0.883). The average IgG and IgA antibody titres in both plasma and saliva of the VF3 + 3 immunized sheep were four to nine times higher than those immunized with VF7 + 3 (P< 0.001) at both 3 and 6 weeks post-secondary immunization. Data also revealed that SF6 methane estimates were consistently higher than the respiration chamber estimates and that there was no significant correlation between the SF6 methane estimates and the respiration chamber methane estimates (R2 = 0.11). PMID:15364447

  12. Transformation of phenol into phenylalanine by a methanogenic consortium

    SciTech Connect

    Lepine, F.; Milot, S.; Beaudet, R.; Villemur, R.

    1996-03-01

    Phenol is a widely used chemical found in many wastewaters of industrial origin. The degradation of phenol by methanogenic bacterial consortia has been reported by many investigators. To better characterise the metabolism of this consortium, a new metabolic pathway of benzoic acid, an intermediary in the degradation of phenol, is reported. This study describes the transformations of benzoic acid into 3-phenylpropionic acid and phenylalanine. 25 refs., 5 figs.

  13. An Intertwined Evolutionary History of Methanogenic Archaea and Sulfate Reduction

    PubMed Central

    Susanti, Dwi; Mukhopadhyay, Biswarup

    2012-01-01

    Hydrogenotrophic methanogenesis and dissimilatory sulfate reduction, two of the oldest energy conserving respiratory systems on Earth, apparently could not have evolved in the same host, as sulfite, an intermediate of sulfate reduction, inhibits methanogenesis. However, certain methanogenic archaea metabolize sulfite employing a deazaflavin cofactor (F420)-dependent sulfite reductase (Fsr) where N- and C-terminal halves (Fsr-N and Fsr-C) are homologs of F420H2 dehydrogenase and dissimilatory sulfite reductase (Dsr), respectively. From genome analysis we found that Fsr was likely assembled from freestanding Fsr-N homologs and Dsr-like proteins (Dsr-LP), both being abundant in methanogens. Dsr-LPs fell into two groups defined by following sequence features: Group I (simplest), carrying a coupled siroheme-[Fe4-S4] cluster and sulfite-binding Arg/Lys residues; Group III (most complex), with group I features, a Dsr-type peripheral [Fe4-S4] cluster and an additional [Fe4-S4] cluster. Group II Dsr-LPs with group I features and a Dsr-type peripheral [Fe4-S4] cluster were proposed as evolutionary intermediates. Group III is the precursor of Fsr-C. The freestanding Fsr-N homologs serve as F420H2 dehydrogenase unit of a putative novel glutamate synthase, previously described membrane-bound electron transport system in methanogens and of assimilatory type sulfite reductases in certain haloarchaea. Among archaea, only methanogens carried Dsr-LPs. They also possessed homologs of sulfate activation and reduction enzymes. This suggested a shared evolutionary history for methanogenesis and sulfate reduction, and Dsr-LPs could have been the source of the oldest (3.47-Gyr ago) biologically produced sulfide deposit. PMID:23028926

  14. Genome sequencing of methanogenic Archaea Methanosarcina mazei TUC01 strain isolated from an Amazonian Flooded Area

    NASA Astrophysics Data System (ADS)

    Baraúna, R. A.; Graças, D. A.; Ramos, R. T.; Carneiro, A. R.; Lopes, T. S.; Lima, A. R.; Zahlouth, R. L.; Pellizari, V. H.; Silva, A.

    2013-05-01

    Methanosarcina mazei is a strictly anaerobic methanogen from the Methanosarcinales order. This species is known for its broad catabolic range among methanogens and is widespread throughout diverse environments. The draft genome of a strain cultivated from the sediment of the Tucuruí hydroelectric power station, the fourth largest hydroelectric dam in the world, is described here. Approximately 80% of methane is produced by biogenic sources, such as methanogenic archaea from M. mazei species. Although the methanogenesis pathway is well known, some aspects of the core genome, genome evolution and shared genes are still unclear. A sediment sample from the Tucuruí hydropower station reservoir was inoculated in mineral media supplemented with acetate and methanol. This media was maintained in an H2:CO2 (80:20) atmosphere to enrich and cultivate M. mazei. The enrichment was conducted at 30°C under standard anaerobic conditions. After several molecular and cellular analyses, total DNA was extracted from a non-pure culture of M. mazei, amplified using phi29 DNA polymerase (BioLabs) and finally used as a source template for genome sequencing. The draft genome was obtained after two rounds of sequencing. First, the genome was sequenced using a SOLiD System V3 with a mate-paired library, which yielded 24,405,103 and 24,399,268 reads (50 bp) for the R3 and F3 tags, respectively. The second round of sequencing was performed using the SOLiD 5500 XL platform with a mate-paired library, resulting in a total of 113,588,848 reads (60 bp) for each tag (F3 and R3). All reads obtained by this procedure were filtered using Quality Assessment software, whereby reads with an average quality score below Phred 20 were removed. Velvet and Edena were used to assemble the reads, and Simplifier was used to remove the redundant sequences. After this, a total of 16,811 contigs were obtained. M. mazei GO1 (AE008384) genome was used to map the contigs and generate the scaffolds. We used the

  15. Hydrogen consumption by methanogens on the early Earth

    NASA Technical Reports Server (NTRS)

    Kral, T. A.; Brink, K. M.; Miller, S. L.; McKay, C. P.; Bada, J. L. (Principal Investigator)

    1998-01-01

    It is possible that the first autotroph used chemical energy rather than light. This could have been the main source of primary production after the initial inventory of abiotic organic material had been depleted. The electron acceptor most readily available for use by this first chemoautotroph would have been CO2. The most abundant electron donor may have been H2 that would have been outgassing from volcanoes at a rate estimated to be as large as 10(12) moles yr-1, as well as from photo-oxidation of Fe+2. We report here that certain methanogens will consume H2 down to partial pressures as low as 4 Pa (4 x 10(-5) atm) with CO2 as the sole carbon source at a rate of 0.7 ng H2 min-1 microgram-1 cell protein. The lower limit of pH2 for growth of methanogens can be understood on the basis that the pH2 needs to be high enough for one ATP to be synthesized per CO2 reduced. The pH2 values needed for growth measured here are consistent with those measured by Stevens and McKinley for growth of methanogens in deep basalt aquifers. H2-consuming autotrophs are likely to have had a profound effect on the chemistry of the early atmosphere and to have been a dominant sink for H2 on the early Earth after life began rather than escape from the Earth's atmosphere to space.

  16. Environmental selection of planktonic methanogens in permafrost thaw ponds.

    PubMed

    Crevecoeur, Sophie; Vincent, Warwick F; Lovejoy, Connie

    2016-01-01

    The warming and thermal erosion of ice-containing permafrost results in thaw ponds that are strong emitters of methane to the atmosphere. Here we examined methanogens and other Archaea, in two types of thaw ponds that are formed by the collapse of either permafrost peat mounds (palsas) or mineral soil mounds (lithalsas) in subarctic Quebec, Canada. Using high-throughput sequencing of a hypervariable region of 16S rRNA, we determined the taxonomic structure and diversity of archaeal communities in near-bottom water samples, and analyzed the mcrA gene transcripts from two sites. The ponds at all sites were well stratified, with hypoxic or anoxic bottom waters. Their archaeal communities were dominated by Euryarchaeota, specifically taxa in the methanogenic orders Methanomicrobiales and Methanosarcinales, indicating a potentially active community of planktonic methanogens. The order Methanomicrobiales accounted for most of the mcrA transcripts in the two ponds. The Archaeal communities differed significantly between the lithalsa and palsa ponds, with higher alpha diversity in the organic-rich palsa ponds, and pronounced differences in community structure. These results indicate the widespread occurrence of planktonic, methane-producing Archaea in thaw ponds, with environmental selection of taxa according to permafrost landscape type. PMID:27501855

  17. Kinetics of chlorinated ethylene dehalogenation under methanogenic conditions

    SciTech Connect

    Skeen, R.S.; Gao, J.; Hooker, B.S.

    1995-12-20

    Kinetics were determined for methanogenic activity and chlorinated ethylene dehalogenation by a methanol-enriched, anaerobic sediment consortium. The culture reductively dechlorinated perchloroethylene (PCE) to trichloroethylene (TCE), 1,1-dichloroethylene (1,1-DCE), vinylchloride (VC), and ethylene and ethane. The absence of methanol or the addition of 2-bromoethanesulfonic acid in the presence of methanol suppressed both methanogenic activity and dechlorination. In contrast, acetate production continued in the presence of 2-bromoethanesulfonic acid. These results suggest that dechlorination was strongly linked to methane formation and not to acetate production. A kinetic model, developed to describe both methanogenesis and dechlorination, successfully predicted experimentally measured concentrations of biomass, methane, substrate, and chlorinated ethylenes. The average maximum specific dehalogenation rates for PCE, TCE, 1,1-DCE, and VC were 0.9 {+-} 0.6, 0.4 {+-} 0.1, 12 {+-} 0.1, and 2.5 {+-} 1.7 {micro}mol contaminant/g {center_dot} DW/day, respectively. This pattern for dechlorination rates is distinctly different than that reported for transition metal cofactors, where rates drop by approximately one order of magnitude as each successive chlorine is removed. The experimental results and kinetic analysis suggest that it will be impractical to targeting methanol consuming methanogenic organisms for in-situ groundwater restoration.

  18. Fast pyrolysis product distribution of biopretreated corn stalk by methanogen.

    PubMed

    Wang, Tipeng; Ye, Xiaoning; Yin, Jun; Jin, Zaixing; Lu, Qiang; Zheng, Zongming; Dong, Changqing

    2014-10-01

    After pretreated by methanogen for 5, 15 and 25 days, corn stalk (CS) were pyrolyzed at 250, 300, 350, 400, 450 and 500 °C by Py-GC/MS and product distribution in bio-oil was analyzed. Results indicated that methanogen pretreatment changed considerably the product distribution: the contents of sugar and phenols increased; the contents of linear carbonyls and furans decreased; the contents of linear ketones and linear acids changed slightly. Methanogen pretreatment improved significantly the pyrolysis selectivity of CS to phenols especially 4-VP. At 250 °C, the phenols content increased from 42.25% for untreated CS to 79.32% for biopretreated CS for 5 days; the 4-VP content increased from 28.6% to 60.9%. Increasing temperature was contributed to convert more lignin into 4-VP, but decreased its content in bio-oil due to more other chemicals formed. The effects of biopretreatment time on the chemicals contents were insignificant. PMID:25113883

  19. Environmental selection of planktonic methanogens in permafrost thaw ponds

    PubMed Central

    Crevecoeur, Sophie; Vincent, Warwick F.; Lovejoy, Connie

    2016-01-01

    The warming and thermal erosion of ice-containing permafrost results in thaw ponds that are strong emitters of methane to the atmosphere. Here we examined methanogens and other Archaea, in two types of thaw ponds that are formed by the collapse of either permafrost peat mounds (palsas) or mineral soil mounds (lithalsas) in subarctic Quebec, Canada. Using high-throughput sequencing of a hypervariable region of 16S rRNA, we determined the taxonomic structure and diversity of archaeal communities in near-bottom water samples, and analyzed the mcrA gene transcripts from two sites. The ponds at all sites were well stratified, with hypoxic or anoxic bottom waters. Their archaeal communities were dominated by Euryarchaeota, specifically taxa in the methanogenic orders Methanomicrobiales and Methanosarcinales, indicating a potentially active community of planktonic methanogens. The order Methanomicrobiales accounted for most of the mcrA transcripts in the two ponds. The Archaeal communities differed significantly between the lithalsa and palsa ponds, with higher alpha diversity in the organic-rich palsa ponds, and pronounced differences in community structure. These results indicate the widespread occurrence of planktonic, methane-producing Archaea in thaw ponds, with environmental selection of taxa according to permafrost landscape type. PMID:27501855

  20. Environmental selection of planktonic methanogens in permafrost thaw ponds

    NASA Astrophysics Data System (ADS)

    Crevecoeur, Sophie; Vincent, Warwick F.; Lovejoy, Connie

    2016-08-01

    The warming and thermal erosion of ice-containing permafrost results in thaw ponds that are strong emitters of methane to the atmosphere. Here we examined methanogens and other Archaea, in two types of thaw ponds that are formed by the collapse of either permafrost peat mounds (palsas) or mineral soil mounds (lithalsas) in subarctic Quebec, Canada. Using high-throughput sequencing of a hypervariable region of 16S rRNA, we determined the taxonomic structure and diversity of archaeal communities in near-bottom water samples, and analyzed the mcrA gene transcripts from two sites. The ponds at all sites were well stratified, with hypoxic or anoxic bottom waters. Their archaeal communities were dominated by Euryarchaeota, specifically taxa in the methanogenic orders Methanomicrobiales and Methanosarcinales, indicating a potentially active community of planktonic methanogens. The order Methanomicrobiales accounted for most of the mcrA transcripts in the two ponds. The Archaeal communities differed significantly between the lithalsa and palsa ponds, with higher alpha diversity in the organic-rich palsa ponds, and pronounced differences in community structure. These results indicate the widespread occurrence of planktonic, methane-producing Archaea in thaw ponds, with environmental selection of taxa according to permafrost landscape type.

  1. Draft Genome Sequence of Bacillus pumilus Fairview, an Isolate Recovered from a Microbial Methanogenic Enrichment of Coal Seam Gas Formation Water from Queensland, Australia

    PubMed Central

    Greenfield, Paul; Tran-Dinh, Nai; Midgley, David J.

    2014-01-01

    Despite its global abundance, Bacillus pumilus is poorly studied. The Fairview strain was obtained from a methanogenic anaerobic coal digester. The draft genome sequence was 3.8 Mbp long and contained 3,890 protein-coding genes. Like the SAFR-032 strain, it includes B. pumilus-specific proteins that likely confer enhanced resistance to environmental stresses. PMID:24744330

  2. Methane production potentials, pathways, and communities of methanogens in vertical sediment profiles of river Sitka

    PubMed Central

    Mach, Václav; Blaser, Martin B.; Claus, Peter; Chaudhary, Prem P.; Rulík, Martin

    2015-01-01

    Biological methanogenesis is linked to permanent water logged systems, e.g., rice field soils or lake sediments. In these systems the methanogenic community as well as the pathway of methane formation are well-described. By contrast, the methanogenic potential of river sediments is so far not well-investigated. Therefore, we analyzed (a) the methanogenic potential (incubation experiments), (b) the pathway of methane production (stable carbon isotopes and inhibitor studies), and (c) the methanogenic community composition (terminal restriction length polymorphism of mcrA) in depth profiles of sediment cores of River Sitka, Czech Republic. We found two depth-related distinct maxima for the methanogenic potentials (a) The pathway of methane production was dominated by hydrogenotrophic methanogenesis (b) The methanogenic community composition was similar in all depth layers (c) The main TRFs were representative for Methanosarcina, Methanosaeta, Methanobacterium, and Methanomicrobium species. The isotopic signals of acetate indicated a relative high contribution of chemolithotrophic acetogenesis to the acetate pool. PMID:26052322

  3. Effects of nitrate dosing on sulfidogenic and methanogenic activities in sewer sediment.

    PubMed

    Liu, Yiwen; Sharma, Keshab R; Ni, Bing-Jie; Fan, Lu; Murthy, Sudhir; Tyson, Gene Q; Yuan, Zhiguo

    2015-05-01

    Nitrate dosing is widely used to control sulfide and methane formation in sewers. The impact of nitrate on sulfide and methane production by sewer biofilms in rising mains has been elucidated recently. However, little is known about the effect of nitrate on biologically active sewer sediment, which is substantially thicker than rising main biofilms (centimeters vs. hundreds of micrometers, respectively). In this study, we investigated the effect of nitrate addition to sewer sediment cultivated in lab-scale sewer sediment reactors. Batch test results showed that nitrate addition does not suppress sulfide production in sewer sediment, but it reduces sulfide accumulation through anoxic sulfide oxidation in the sediment and hence, also reduces sulfide accumulation in the bulk water. Microsensor measurement of sediment sulfide revealed the presence of sulfide oxidation and sulfide production zones with the interface dynamically regulated by the depth of nitrate penetration. In contrast, the methane production activity of sewer sediment was substantially reduced, likely due to the long-term inhibitory effects of nitrate on methanogens. Pore water measurements showed that methane production activity in the sediment zone with frequent nitrate exposure was completely suppressed, and consequently, the methane production zone re-established deeper in the sediment where nitrate penetration was infrequent. PMID:25727155

  4. A phytoene desaturase homolog gene from the methanogenic archaeon Methanosarcina acetivorans is responsible for hydroxyarchaeol biosynthesis.

    PubMed

    Mori, Takeshi; Isobe, Keisuke; Ogawa, Takuya; Yoshimura, Tohru; Hemmi, Hisashi

    2015-10-16

    Hydroxyarchaeols are the typical core structures of archaeal membrane lipids uniquely produced by a limited number of methanogenic lineages, which are mainly classified in orders Methanosarcinales and Methanococcales. However, the biosynthetic machinery that is used for the biosynthesis of hydroxyarcheol core lipids has not been discovered. In this study, the ma0127 gene from Methanosarcina acetivorans, which encodes a phytoene desaturase-like protein, was found to be responsible for the hydration of a geranylgeranyl group in an archaeal-lipid precursor, sn-2,3-O-digeranylgeranylglyceryl phosphoglycerol, produced in Escherichia coli cells expressing several archaeal enzymes. LC-ESI-tandem-MS analyses proved that hydration occurs at the 2',3'-double bond of the geranylgeranyl group, yielding a 3'-hydroxylated lipid precursor. This result suggests that the encoded protein MA0127 is a hydratase involved in hydroxyarchaeol biosynthesis, because M. acetivorans is known to produce hydroxyarchaeol core lipids with a 3'-hydroxyphytanyl group. Furthermore, the distribution of the putative orthologs of ma0127 among methanogens is generally in good agreement with that of hydroxyarchaeol producers, including anaerobic methanotrophs (ANMEs). PMID:26361140

  5. Physiological and Transcriptomic Analyses of the Thermophilic, Aceticlastic Methanogen Methanosaeta thermophila Responding to Ammonia Stress

    PubMed Central

    Kato, Souichiro; Sasaki, Konomi; Watanabe, Kazuya; Yumoto, Isao; Kamagata, Yoichi

    2014-01-01

    The inhibitory effects of ammonia on two different degradation pathways of methanogenic acetate were evaluated using a pure culture (Methanosaeta thermophila strain PT) and defined co-culture (Methanothermobacter thermautotrophicus strain TM and Thermacetogenium phaeum strain PB), which represented aceticlastic and syntrophic methanogenesis, respectively. Growth experiments with high concentrations of ammonia clearly demonstrated that sensitivity to ammonia stress was markedly higher in M. thermophila PT than in the syntrophic co-culture. M. thermophila PT also exhibited higher sensitivity to high pH stress, which indicated that an inability to maintain pH homeostasis is an underlying cause of ammonia inhibition. Methanogenesis was inhibited in the resting cells of M. thermophila PT with moderate concentrations of ammonia, suggesting that the inhibition of enzymes involved in methanogenesis may be one of the major factors responsible for ammonia toxicity. Transcriptomic analysis revealed a broad range of disturbances in M. thermophila PT cells under ammonia stress conditions, including protein denaturation, oxidative stress, and intracellular cation imbalances. The results of the present study clearly demonstrated that syntrophic acetate degradation dominated over aceticlastic methanogenesis under ammonia stress conditions, which is consistent with the findings of previous studies on complex microbial community systems. Our results also imply that the co-existence of multiple metabolic pathways and their different sensitivities to stress factors confer resiliency on methanogenic processes. PMID:24920170

  6. Energetic and hydrogen limitations of thermophilic and hyperthermophilic methanogens

    NASA Astrophysics Data System (ADS)

    Stewart, L. C.; Holden, J. F.

    2013-12-01

    Deep-sea hydrothermal vents are a unique ecosystem, based ultimately not on photosynthesis but chemosynthetic primary production. This makes them an excellent analog environment for the early Earth, and for potential extraterrestrial habitable environments, such as those on Mars and Europa. The habitability of given vent systems for chemoautotrophic prokaryotes can be modeled energetically by estimating the available Gibbs energy for specific modes of chemoautotrophy, using geochemical data and mixing models for hydrothermal fluids and seawater (McCollom and Shock, 1997). However, modeling to date has largely not taken into account variation in organisms' energy demands in these environments. Controls on maintenance energies are widely assumed to be temperature-dependent, rising with increasing temperature optima (Tijhuis et al., 1993), and species-independent. The impacts of other environmental stressors and particular energy-gathering strategies on maintenance energies have not been investigated. We have undertaken culture-based studies of growth and maintenance energies in thermophilic and hyperthermophilic methanogenic (hydrogenotrophic) archaea from deep-sea hydrothermal vents to investigate potential controls on energy demands in hydrothermal vent microbes, and to quantify their growth and maintenance energies for future bioenergetic modeling. We have investigated trends in their growth energies over their full temperature range and a range of nitrogen concentrations, and in their maintenance energies at different hydrogen concentrations. Growth energies in these organisms appear to rise with temperature, but do not vary between hyperthermophilic and thermophilic methanogens. Nitrogen availability at tested levels (40μM - 9.4 mM) does not appear to affect growth energies in all but one tested organism. In continuous chemostat culture, specific methane production varied with hydrogen availability but was similar between a thermophilic and a hyperthermophilic

  7. Estimation of Methanogen Biomass by Quantitation of Coenzyme M

    PubMed Central

    Elias, Dwayne A.; Krumholz, Lee R.; Tanner, Ralph S.; Suflita, Joseph M.

    1999-01-01

    Determination of the role of methanogenic bacteria in an anaerobic ecosystem often requires quantitation of the organisms. Because of the extreme oxygen sensitivity of these organisms and the inherent limitations of cultural techniques, an accurate biomass value is very difficult to obtain. We standardized a simple method for estimating methanogen biomass in a variety of environmental matrices. In this procedure we used the thiol biomarker coenzyme M (CoM) (2-mercaptoethanesulfonic acid), which is known to be present in all methanogenic bacteria. A high-performance liquid chromatography-based method for detecting thiols in pore water (A. Vairavamurthy and M. Mopper, Anal. Chim. Acta 78:363–370, 1990) was modified in order to quantify CoM in pure cultures, sediments, and sewage water samples. The identity of the CoM derivative was verified by using liquid chromatography-mass spectroscopy. The assay was linear for CoM amounts ranging from 2 to 2,000 pmol, and the detection limit was 2 pmol of CoM/ml of sample. CoM was not adsorbed to sediments. The methanogens tested contained an average of 19.5 nmol of CoM/mg of protein and 0.39 ± 0.07 fmol of CoM/cell. Environmental samples contained an average of 0.41 ± 0.17 fmol/cell based on most-probable-number estimates. CoM was extracted by using 1% tri-(N)-butylphosphine in isopropanol. More than 90% of the CoM was recovered from pure cultures and environmental samples. We observed no interference from sediments in the CoM recovery process, and the method could be completed aerobically within 3 h. Freezing sediment samples resulted in 46 to 83% decreases in the amounts of detectable CoM, whereas freezing had no effect on the amounts of CoM determined in pure cultures. The method described here provides a quick and relatively simple way to estimate methanogenic biomass. PMID:10584015

  8. Biological hydrogen production using chloroform-treated methanogenic granules.

    PubMed

    Hu, Bo; Chen, Shulin

    2008-03-01

    In fermentative hydrogen production, the low-hydrogen-producing bacteria retention rate limits the suspended growth reactor productivity because of the long hydraulic retention time (HRT) required to maintain adequate bacteria population. Traditional bacteria immobilization methods such as calcium alginate entrapment have many application limitations in hydrogen fermentation, including limited duration time, bacteria leakage, cost, and so on. The use of chloroform-treated anaerobic granular sludge as immobilized hydrogen-producing bacteria in an immobilized hydrogen culture may be able to overcome the limitations of traditional immobilization methods. This paper reports the findings on the performance of fed-batch cultures and continuous cultures inoculated with chloroform-treated granules. The chloroform-treated granules were able to be reused over four fed-batch cultures, with pH adjustment. The upflow reactor packed with chloroform-treated granules was studied, and the HRT of the upflow reactor was found to be as low as 4 h without any decrease in hydrogen production yield. Initial pH and glucose concentration of the culture medium significantly influenced the performance of the reactor. The optimum initial pH of the culture medium was neutral, and the optimum glucose concentration of the culture medium was below 20 g chemical oxygen demand/L at HRT 4 h. This study also investigated the possibility of integrating immobilized hydrogen fermentation using chloroform-treated granules with immobilized methane production using untreated granular sludge. The results showed that the integrated batch cultures produced 1.01 mol hydrogen and 2 mol methane per mol glucose. Treating the methanogenic granules with chloroform and then using the treated granules as immobilized hydrogen-producing sludge demonstrated advantages over other immobilization methods because the treated granules provide hydrogen-producing bacteria with a protective niche, a long duration of an active

  9. Biological Hydrogen Production Using Chloroform-treated Methanogenic Granules

    NASA Astrophysics Data System (ADS)

    Hu, Bo; Chen, Shulin

    In fermentative hydrogen production, the low-hydrogen-producing bacteria retention rate limits the suspended growth reactor productivity because of the long hydraulic retention time (HRT) required to maintain adequate bacteria population. Traditional bacteria immobilization methods such as calcium alginate entrapment have many application limitations in hydrogen fermentation, including limited duration time, bacteria leakage, cost, and so on. The use of chloroform-treated anaerobic granular sludge as immobilized hydrogen-producing bacteria in an immobilized hydrogen culture may be able to overcome the limitations of traditional immobilization methods. This paper reports the findings on the performance of fed-batch cultures and continuous cultures inoculated with chloroform-treated granules. The chloroform-treated granules were able to be reused over four fed-batch cultures, with pH adjustment. The upflow reactor packed with chloroform-treated granules was studied, and the HRT of the upflow reactor was found to be as low as 4 h without any decrease in hydrogen production yield. Initial pH and glucose concentration of the culture medium significantly influenced the performance of the reactor. The optimum initial pH of the culture medium was neutral, and the optimum glucose concentration of the culture medium was below 20 g chemical oxygen demand/L at HRT 4 h. This study also investigated the possibility of integrating immobilized hydrogen fermentation using chloroform-treated granules with immobilized methane production using untreated granular sludge. The results showed that the integrated batch cultures produced 1.01 mol hydrogen and 2 mol methane per mol glucose. Treating the methanogenic granules with chloroform and then using the treated granules as immobilized hydrogen-producing sludge demonstrated advantages over other immobilization methods because the treated granules provide hydrogen-producing bacteria with a protective niche, a long duration of an active

  10. Effects of triclosan, diclofenac, and nonylphenol on mesophilic and thermophilic methanogenic activity and on the methanogenic communities.

    PubMed

    Symsaris, Evangelos C; Fotidis, Ioannis A; Stasinakis, Athanasios S; Angelidaki, Irini

    2015-06-30

    In this study, a toxicity assay using a mesophilic wastewater treatment plant sludge-based (SI) and a thermophilic manure-based inoculum (MI), under different biomass concentrations was performed to define the effects of diclofenac (DCF), triclosan (TCS), and nonylphenol (NP) on anaerobic digestion (AD) process. Additionally, the influence of DCF, TCS, and NP on the relative abundance of the methanogenic populations was investigated. Results obtained demonstrated that, in terms of methane production, SI inoculum was more resistant to the toxicity effect of DCF, TCS, and NP, compared to the MI inoculum. The IC50 values were 546, 35, and 363 mg L(-1) for SI inoculum and 481, 32, and 74 mg L(-1) for MI inoculum for DCF, TCS, and NP, respectively. For both inocula, higher biomass concentrations reduced the toxic effect of TCS (higher methane production up to 64%), contrary to DCF, where higher biomass loads decreased methane yield up to 31%. Fluorescence in situ hybridization analysis showed that hydrogenotrophic methanogens were more resistant to the inhibitory effect of DCF, TCS, and NP compared to aceticlastic methanogens. PMID:25768988

  11. Differences in the Rumen Methanogen Populations of Lactating Jersey and Holstein Dairy Cows under the Same Diet Regimen▿†

    PubMed Central

    King, Erin E.; Smith, Rachel P.; St-Pierre, Benoit; Wright, André-Denis G.

    2011-01-01

    In the dairy cattle industry, Holstein and Jersey are the breeds most commonly used for production. They differ in performance by various traits, such as body size, milk production, and milk composition. With increased concerns about the impact of agriculture on climate change, potential differences in other traits, such as methane emission, also need to be characterized further. Since methane is produced in the rumen by methanogenic archaea, we investigated whether the population structure of methanogen communities would differ between Holsteins and Jerseys. Breed-specific rumen methanogen 16S rRNA gene clone libraries were constructed from pooled PCR products obtained from lactating Holstein and Jersey cows, generating 180 and 185 clones, respectively. The combined 365 sequences were assigned to 55 species-level operational taxonomic units (OTUs). Twenty OTUs, representing 85% of the combined library sequences, were common to both breeds, while 23 OTUs (36 sequences) were found only in the Holstein library and 12 OTUs (18 sequences) were found only in the Jersey library, highlighting increased diversity in the Holstein library. Other differences included the observation that sequences with species-like sequence identity to Methanobrevibacter millerae were represented more highly in the Jersey breed, while Methanosphaera-related sequences and novel uncultured methanogen clones were more frequent in the Holstein library. In contrast, OTU sequences with species-level sequence identity to Methanobrevibacter ruminantium were represented similarly in both libraries. Since the sampled animals were from a single herd consisting of two breeds which were fed the same diet and maintained under the same environmental conditions, the differences we observed may be due to differences in host breed genetics. PMID:21705541

  12. Shifts in methanogenic community composition and methane fluxes along the degradation of discontinuous permafrost

    PubMed Central

    Liebner, Susanne; Ganzert, Lars; Kiss, Andrea; Yang, Sizhong; Wagner, Dirk; Svenning, Mette M.

    2015-01-01

    The response of methanogens to thawing permafrost is an important factor for the global greenhouse gas budget. We tracked methanogenic community structure, activity, and abundance along the degradation of sub-Arctic palsa peatland permafrost. We observed the development of pronounced methane production, release, and abundance of functional (mcrA) methanogenic gene numbers following the transitions from permafrost (palsa) to thaw pond structures. This was associated with the establishment of a methanogenic community consisting both of hydrogenotrophic (Methanobacterium, Methanocellales), and potential acetoclastic (Methanosarcina) members and their activity. While peat bog development was not reflected in significant changes of mcrA copy numbers, potential methane production, and rates of methane release decreased. This was primarily linked to a decline of potential acetoclastic in favor of hydrogenotrophic methanogens. Although palsa peatland succession offers similarities with typical transitions from fen to bog ecosystems, the observed dynamics in methane fluxes and methanogenic communities are primarily attributed to changes within the dominant Bryophyta and Cyperaceae taxa rather than to changes in peat moss and sedge coverage, pH and nutrient regime. Overall, the palsa peatland methanogenic community was characterized by a few dominant operational taxonomic units (OTUs). These OTUs seem to be indicative for methanogenic species that thrive in terrestrial organic rich environments. In summary, our study shows that after an initial stage of high methane emissions following permafrost thaw, methane fluxes, and methanogenic communities establish that are typical for northern peat bogs. PMID:26029170

  13. Shifts in methanogenic community composition and methane fluxes along the degradation of discontinuous permafrost.

    PubMed

    Liebner, Susanne; Ganzert, Lars; Kiss, Andrea; Yang, Sizhong; Wagner, Dirk; Svenning, Mette M

    2015-01-01

    The response of methanogens to thawing permafrost is an important factor for the global greenhouse gas budget. We tracked methanogenic community structure, activity, and abundance along the degradation of sub-Arctic palsa peatland permafrost. We observed the development of pronounced methane production, release, and abundance of functional (mcrA) methanogenic gene numbers following the transitions from permafrost (palsa) to thaw pond structures. This was associated with the establishment of a methanogenic community consisting both of hydrogenotrophic (Methanobacterium, Methanocellales), and potential acetoclastic (Methanosarcina) members and their activity. While peat bog development was not reflected in significant changes of mcrA copy numbers, potential methane production, and rates of methane release decreased. This was primarily linked to a decline of potential acetoclastic in favor of hydrogenotrophic methanogens. Although palsa peatland succession offers similarities with typical transitions from fen to bog ecosystems, the observed dynamics in methane fluxes and methanogenic communities are primarily attributed to changes within the dominant Bryophyta and Cyperaceae taxa rather than to changes in peat moss and sedge coverage, pH and nutrient regime. Overall, the palsa peatland methanogenic community was characterized by a few dominant operational taxonomic units (OTUs). These OTUs seem to be indicative for methanogenic species that thrive in terrestrial organic rich environments. In summary, our study shows that after an initial stage of high methane emissions following permafrost thaw, methane fluxes, and methanogenic communities establish that are typical for northern peat bogs. PMID:26029170

  14. Hydrogen consumption by methanogens on the early Earth.

    PubMed

    Kral, T A; Brink, K M; Miller, S L; McKay, C P

    1998-06-01

    It is possible that the first autotroph used chemical energy rather than light. This could have been the main source of primary production after the initial inventory of abiotic organic material had been depleted. The electron acceptor most readily available for use by this first chemoautotroph would have been CO2. The most abundant electron donor may have been H2 that would have been outgassing from volcanoes at a rate estimated to be as large as 10(12) moles yr-1, as well as from photo-oxidation of Fe+2. We report here that certain methanogens will consume H2 down to partial pressures as low as 4 Pa (4 x 10(-5) atm) with CO2 as the sole carbon source at a rate of 0.7 ng H2 min-1 microgram-1 cell protein. The lower limit of pH2 for growth of methanogens can be understood on the basis that the pH2 needs to be high enough for one ATP to be synthesized per CO2 reduced. The pH2 values needed for growth measured here are consistent with those measured by Stevens and McKinley for growth of methanogens in deep basalt aquifers. H2-consuming autotrophs are likely to have had a profound effect on the chemistry of the early atmosphere and to have been a dominant sink for H2 on the early Earth after life began rather than escape from the Earth's atmosphere to space. PMID:9611769

  15. Kinetics of phenol biodegradation by an immobilized methanogenic consortium

    SciTech Connect

    Dwyer, D.F.; Krumme, M.L.; Boyd, S.A.; Tiedje, J.M.

    1986-08-01

    A phenol-degrading methanogenic enrichment was successfully immobilized in agar as shown by the stoichiometric conversion of phenol to CH/sub 4/ and CO/sub 2/. The enrichment contained members of three physiological groups necessary for the syntrophic mineralization of phenol: a phenol-oxidizing bacterium, a Methanothrix-like bacterium, and an H/sub 2/-utilizing methanogen. The immobilization technique resulted in the cells being embedded in a long, thin agar strans (1 mm in diameter by 2 to 50 cm in length) that resembled spaghetti. Immobilization had three effects as shown by a comparative kinetic analysis of phenol degradation by free versus immobilized cells. (1) The maximum rate of degradation was reduced from 14.8 to 10.0 ..mu..g of phenol per h; (2) the apparent K/sub m/ for the overall reaction was reduced from 90 to 46 ..mu..g of phenol per ml. probably because of the retention of acetate, H/sub 2/ and CO/sub 2/ in the proximity of immobilized methanogens; and (3) the cells were protected from substrate inhibition caused by high concentrations of phenol, which increased the apparent K/sub i/ value from 900 to 1725 ..mu..g of phenol per ml. Estimates for the kinetic parameters K/sub m/, K/sub i/, and V/sub max/ were used in a modified substrate inhibition model that simulated rates of phenol degradation for given phenol concentrations. The simulated rates were in close agreement with experimentally derived rates for both stimulatory and inhibitory concentrations of phenol.

  16. Degradation of hydrocarbons under methanogenic conditions in different geosystems

    NASA Astrophysics Data System (ADS)

    Straaten, Nontje; Jiménez García, Núria; Richnow, Hans-Hermann; Krueger, Martin

    2014-05-01

    With increasing energy demand the search for new resources is becoming increasingly important for the future energy supply. Therefore the knowledge about fossil fuels like oil or natural gas and their extraction should be expanded. Biodegraded oil is found in many reservoirs worldwide. Consequently, it is very important to get insight in the microbial communities and metabolic processes involved in hydrocarbon degradation. Due to the lack of alternative electron acceptors in hydrocarbon-rich geosystems, degradation often takes place under methanogenic conditions. The aim of the present study is to identify the microorganisms and mechanisms involved in the degradation of complex hydrocarbons, like BTEX and polycyclic aromatic hydrocarbons, using culture dependent and independent techniques. For this purpose enrichment cultures from marine sediments, shales, coal and oil reservoirs are monitored for their capability to degrade alkanes and aromatic compounds. Moreover the environmental samples of these different geosystems analysed for evidence for the in situ occurrence of methanogenic oil degradation. The gas geochemical data provided in several cases hints for a recent biological origin of the methane present. First results of the microbial community analysis showed in environmental samples and enrichment cultures the existence of Bacteria known to degrade hydrocarbons. Also a diverse community of methanogenic Archaea could be found in the clone libraries. Additionally, in oil and coal reservoir samples the degradation of model hydrocarbons, e.g. methylnaphthalene, hexadecane and BTEX, to CH4 was confirmed by 13C-labeling. To explore the mechanisms involved in biodegradation, the enrichments as well as the original environmental samples are further analysed for the presence of respective functional genes.

  17. Effect of temperature on perchloroethylene dechlorination by a methanogenic consortium

    SciTech Connect

    Gao, J.; Skeen, R.S.; Hooker, B.S.

    1995-04-01

    The effect of temperature on the kinetics of growth, substrate metabolism, and perchloroethylene (PCE) dechlorination by a methanogenic consortium is reported. In all cases, a simple kinetic model accurately reflected experimental data. Values for the substrate and methane yield coefficients, and the maximum specific growth rate are fairly consistent at each temperature. Also, the substrate and methane yield coefficients show little temperature sensitivity. In contrast, both the maximum specific growth rate and the PCE dechlorination yield coefficient (Y{sub PCE}) are temperature dependent.

  18. Elucidation of methanogenic coenzyme biosyntheses: from spectroscopy to genomics.

    PubMed

    Graham, David E; White, Robert H

    2002-04-01

    Methanogenesis, the anaerobic production of methane from CO2 or simple carbon compounds, requires seven organic coenzymes. This review describes pathways for the biosynthesis of methanofuran, 5,6,7,8-tetrahydromethanopterin, coenzyme F420, coenzyme M (2-mercaptoethanesulfonic acid) and coenzyme B (7-mercaptoheptanoyl-L-threonine phosphate). Spectroscopic evidence for the pathways is reviewed and recent efforts are described to identify and characterize the biosynthetic enzymes from methanogenic archaea. The literature from 1971 to September 2001 is reviewed, and 169 references are cited. PMID:12013276

  19. Anaerobic biodegradation of cyanide under methanogenic conditions.

    PubMed Central

    Fallon, R D; Cooper, D A; Speece, R; Henson, M

    1991-01-01

    Upflow, anaerobic, fixed-bed, activated charcoal biotreatment columns capable of operating at free cyanide concentrations of greater than 100 mg liter-1 with a hydraulic retention time of less than 48 h were developed. Methanogenesis was maintained under a variety of feed medium conditions which included ethanol, phenol, or methanol as the primary reduced carbon source. Under optimal conditions, greater than 70% of the inflow free cyanide was removed in the first 30% of the column height. Strongly complexed cyanides were resistant to removal. Ammonia was the nitrogen end product of cyanide transformation. In cell material removed from the charcoal columns, [14C]bicarbonate was the major carbon end product of [14C]cyanide transformation. PMID:1872600

  20. Methane Production and Methanogenic Archaea in the Digestive Tracts of Millipedes (Diplopoda)

    PubMed Central

    Šustr, Vladimír; Chroňáková, Alica; Semanová, Stanislava; Tajovský, Karel; Šimek, Miloslav

    2014-01-01

    Methane production by intestinal methanogenic Archaea and their community structure were compared among phylogenetic lineages of millipedes. Tropical and temperate millipedes of 35 species and 17 families were investigated. Species that emitted methane were mostly in the juliform orders Julida, Spirobolida, and Spirostreptida. The irregular phylogenetic distribution of methane production correlated with the presence of the methanogen-specific mcrA gene. The study brings the first detailed survey of methanogens’ diversity in the digestive tract of millipedes. Sequences related to Methanosarcinales, Methanobacteriales, Methanomicrobiales and some unclassified Archaea were detected using molecular profiling (DGGE). The differences in substrate preferences of the main lineages of methanogenic Archaea found in different millipede orders indicate that the composition of methanogen communities may reflect the differences in available substrates for methanogenesis or the presence of symbiotic protozoa in the digestive tract. We conclude that differences in methane production in the millipede gut reflect differences in the activity and proliferation of intestinal methanogens rather than an absolute inability of some millipede taxa to host methanogens. This inference was supported by the general presence of methanogenic activity in millipede faecal pellets and the presence of the 16S rRNA gene of methanogens in all tested taxa in the two main groups of millipedes, the Helminthophora and the Pentazonia. PMID:25028969

  1. Draft Genome Sequence of the Rumen Methanogen Methanobrevibacter olleyae YLM1

    PubMed Central

    Kelly, William J.; Li, Dong; Lambie, Suzanne C.; Cox, Faith; Attwood, Graeme T.; Altermann, Eric

    2016-01-01

    Methanobrevibacter olleyae YLM1 is a hydrogenotrophic methanogen, isolated from the rumen of a lamb. Its genome has been sequenced to provide information on the genomic diversity of rumen methanogens and support the development of approaches to reduce methane formation by ruminants. PMID:27056228

  2. Molecular Diversity and Activity of Methanogens in the Subseafloor at Deep-Sea Hydrothermal Vents of the Pacific Ocean (Invited)

    NASA Astrophysics Data System (ADS)

    Huber, J. A.; Merkel, A.; Holden, J. F.; Lilley, M. D.; Butterfield, D. A.

    2009-12-01

    , including mesophiles and hyper/thermophiles, but that some methanogens recovered from Marker 113 are surviving at low or sub-optimal hydrogen levels. Vent 9m had a community composition similar to Marker 113, dominated by Methanococcales, and Zen Gardens, also at Axial, had a population of methanogens very different from either site. The community there was composed of members of the order Methanomicrobiales, including mesophilic methanogens previously only found in terrestrial environments. Along the Mariana Arc, Shrimp City vent at NW Rota-1 had some of the highest levels of methane detected on the entire arc, and mcrA analysis indicates members of the Methanococcales were present, as well as Methanosarcinales and anaerobic methane oxidizers. An integrated comparison of organismal and geochemical diversity will be presented to link energy transfer in these diverse hydrothermal systems from mantle to microbes.

  3. Spirulina cultivation in China

    NASA Astrophysics Data System (ADS)

    Wu, Bo-Tang; Xiang, Wen-Zhou; Zeng, Cheng-Kui

    1998-03-01

    This paper reviews and discusses the development and many problems of Spirulina cultivation in China, points out the advantages and disadvantages of open photobioreactor system, and predicts that seawater Spirulina cultivation will be a new trend to be strengthened and emphasized due to its special physiological characteristics, easier management, lower fertilizer cost, and higher resistance to contaminants and rare pollution of chemicals.

  4. Oligonucleotide primers, probes and molecular methods for the environmental monitoring of methanogenic archaea

    PubMed Central

    Narihiro, Takashi; Sekiguchi, Yuji

    2011-01-01

    Summary For the identification and quantification of methanogenic archaea (methanogens) in environmental samples, various oligonucleotide probes/primers targeting phylogenetic markers of methanogens, such as 16S rRNA, 16S rRNA gene and the gene for the α‐subunit of methyl coenzyme M reductase (mcrA), have been extensively developed and characterized experimentally. These oligonucleotides were designed to resolve different groups of methanogens at different taxonomic levels, and have been widely used as hybridization probes or polymerase chain reaction primers for membrane hybridization, fluorescence in situ hybridization, rRNA cleavage method, gene cloning, DNA microarray and quantitative polymerase chain reaction for studies in environmental and determinative microbiology. In this review, we present a comprehensive list of such oligonucleotide probes/primers, which enable us to determine methanogen populations in an environment quantitatively and hierarchically, with examples of the practical applications of the probes and primers. PMID:21375721

  5. Chloramphenicol acetyltransferase should not provide methanogens with resistance to chloramphenicol. [Methanococcus voltae; Methanococcus vannielii; Methanococcus deltae; Methanobrevibacter smithii

    SciTech Connect

    Beckler, G.S.; Hook, L.A.; Reeve, J.N.

    1984-04-01

    Growth of the four methanogens investigated was inhibitied by chloramphenicol-3-acetate; therefore, introduction of chloramphenicol acetyltransferase-encoding genes should not confer chloramphenicol resistance on these methanogens. Reduction of the aryl nitro group of chloramphenicol produced a compound which did not inhibit the growth of these methanogens. 9 references.

  6. Complete Genome Sequence of Methanogenic Archaeon ISO4-G1, a Member of the Methanomassiliicoccales, Isolated from a Sheep Rumen

    PubMed Central

    Kelly, William J.; Li, Dong; Lambie, Suzanne C.; Jeyanathan, Jeyamalar; Cox, Faith; Li, Yang; Attwood, Graeme T.; Altermann, Eric

    2016-01-01

    Methanogenic archaeon ISO4-G1 is a methylotrophic methanogen belonging to the order Methanomassiliicoccales that was isolated from a sheep rumen. Its genome has been sequenced to provide information on the genetic diversity of rumen methanogens in order to develop technologies for ruminant methane mitigation. PMID:27056226

  7. Methane formation and methane oxidation by methanogenic bacteria.

    PubMed Central

    Zehnder, A J; Brock, T D

    1979-01-01

    Methanogenic bacteria were found to form and oxidize methane at the same time. As compared to the quantity of methane formed, the amount of methane simultaneously oxidized varied between 0.3 and 0.001%, depending on the strain used. All the nine tested strains of methane producers (Methanobacterium ruminantium, Methanobacterium strain M.o.H., M. formicicum, M. thermoautotrophicum, M. arbophilicum, Methanobacterium strain AZ, Methanosarcina barkeri, Methanospirillum hungatii, and the "acetate organism") reoxidized methane to carbon dioxide. In addition, they assimilated a small part of the methane supplied into cell material. Methanol and acetate also occurred as oxidation products in M. barkeri cultures. Acetate was also formed by the "acetate organism," a methane bacterium unable to use methanogenic substrates other than acetate. Methane was the precursor of the methyl group of the acetate synthesized in the course of methane oxidation. Methane formation and its oxidation were inhibited equally by 2-bromoethanesulfonic acid. Short-term labeling experiments with M. thermoautotrophicum and M. hungatii clearly suggest that the pathway of methane oxidation is not identical with a simple back reaction of the methane formation process. Images PMID:762019

  8. Methanogens rapidly transition from methane production to iron reduction.

    PubMed

    Sivan, O; Shusta, S S; Valentine, D L

    2016-03-01

    Methanogenesis, the microbial methane (CH4 ) production, is traditionally thought to anchor the mineralization of organic matter as the ultimate respiratory process in deep sediments, despite the presence of oxidized mineral phases, such as iron oxides. This process is carried out by archaea that have also been shown to be capable of reducing iron in high levels of electron donors such as hydrogen. The current pure culture study demonstrates that methanogenic archaea (Methanosarcina barkeri) rapidly switch from methanogenesis to iron-oxide reduction close to natural conditions, with nitrogen atmosphere, even when faced with substrate limitations. Intensive, biotic iron reduction was observed following the addition of poorly crystalline ferrihydrite and complex organic matter and was accompanied by inhibition of methane production. The reaction rate of this process was of the first order and was dependent only on the initial iron concentrations. Ferrous iron production did not accelerate significantly with the addition of 9,10-anthraquinone-2,6-disulfonate (AQDS) but increased by 11-28% with the addition of phenazine-1-carboxylate (PCA), suggesting the possible role of methanophenazines in the electron transport. The coupling between ferrous iron and methane production has important global implications. The rapid transition from methanogenesis to reduction of iron-oxides close to the natural conditions in sediments may help to explain the globally-distributed phenomena of increasing ferrous concentrations below the traditional iron reduction zone in the deep 'methanogenic' sediment horizon, with implications for metabolic networking in these subsurface ecosystems and in past geological settings. PMID:26762691

  9. Restricted diversity of dental calculus methanogens over five centuries, France.

    PubMed

    Huynh, Hong T T; Nkamga, Vanessa D; Signoli, Michel; Tzortzis, Stéfan; Pinguet, Romuald; Audoly, Gilles; Aboudharam, Gérard; Drancourt, Michel

    2016-01-01

    Methanogens are acknowledged archaeal members of modern dental calculus microbiota and dental pathogen complexes. Their repertoire in ancient dental calculus is poorly known. We therefore investigated archaea in one hundred dental calculus specimens collected from individuals recovered from six archaeological sites in France dated from the 14(th) to 19(th) centuries AD. Dental calculus was demonstrated by macroscopic and cone-beam observations. In 56 calculus specimens free of PCR inhibition, PCR sequencing identified Candidatus Methanobrevibacter sp. N13 in 44.6%, Methanobrevibacter oralis in 19.6%, a new Methanomassiliicoccus luminyensis-like methanogen in 12.5%, a Candidatus Nitrososphaera evergladensis-like in one and Methanoculleus bourgensis in one specimen, respectively. One Candidatus Methanobrevibacter sp. N13 dental calculus was further documented by fluorescent in situ hybridization. The prevalence of dental calculus M. oralis was significantly lower in past populations than in modern populations (P = 0.03, Chi-square test). This investigation revealed a previously unknown repertoire of archaea found in the oral cavity of past French populations as reflected in preserved dental calculus. PMID:27166431

  10. Study of methanogens by genetic techniques. A subcontract progress report

    SciTech Connect

    Baresi, L.; Bertani, G.

    1984-06-01

    Genetic studies of methanogenic bacteria could lead to better exploitation of these organisms in the production of methane from biomass. The objective of this study is to develop a workable genetic system for these bacteria. We have tried to apply standard genetic techniques to these slow growing, strictly anaerobic bacteria. We have been able to isolate several types of mutants both spontaneous and induced. For Methanococcus voltae we have (a) established survival curves to ultraviolet light and gamma ray irradiation, (b) isolated by direct selection mutants resistant to bromo-ethane-sulfonate and to 5-methyl-tryptophan, and a double mutant exhibiting both resistances, (c) isolated after mutagenesis two nutritional mutants (one requiring histidine and the other requiring adenine and possibly another factor). For Methanobacterium thermoautotrophicum we have (a) established an ultraviolet light survival curve, (b) isolated by direct selection mutants resistant to bromo-ethane-sulfonate and to gentamicin. We have developed a routine technique for the purpose of isolating phages capable of infecting methanogenic bacteria. 10 figures, 1 table.

  11. Genome Copy Numbers and Gene Conversion in Methanogenic Archaea▿

    PubMed Central

    Hildenbrand, Catherina; Stock, Tilmann; Lange, Christian; Rother, Michael; Soppa, Jörg

    2011-01-01

    Previous studies revealed that one species of methanogenic archaea, Methanocaldococcus jannaschii, is polyploid, while a second species, Methanothermobacter thermoautotrophicus, is diploid. To further investigate the distribution of ploidy in methanogenic archaea, species of two additional genera—Methanosarcina acetivorans and Methanococcus maripaludis—were investigated. M. acetivorans was found to be polyploid during fast growth (tD = 6 h; 17 genome copies) and oligoploid during slow growth (doubling time = 49 h; 3 genome copies). M. maripaludis has the highest ploidy level found for any archaeal species, with up to 55 genome copies in exponential phase and ca. 30 in stationary phase. A compilation of archaeal species with quantified ploidy levels reveals a clear dichotomy between Euryarchaeota and Crenarchaeota: none of seven euryarchaeal species of six genera is monoploid (haploid), while, in contrast, all six crenarchaeal species of four genera are monoploid, indicating significant genetic differences between these two kingdoms. Polyploidy in asexual species should lead to accumulation of inactivating mutations until the number of intact chromosomes per cell drops to zero (called “Muller's ratchet”). A mechanism to equalize the genome copies, such as gene conversion, would counteract this phenomenon. Making use of a previously constructed heterozygous mutant strain of the polyploid M. maripaludis we could show that in the absence of selection very fast equalization of genomes in M. maripaludis took place probably via a gene conversion mechanism. In addition, it was shown that the velocity of this phenomenon is inversely correlated to the strength of selection. PMID:21097629

  12. Restricted diversity of dental calculus methanogens over five centuries, France

    PubMed Central

    Huynh, Hong T. T.; Nkamga, Vanessa D.; Signoli, Michel; Tzortzis, Stéfan; Pinguet, Romuald; Audoly, Gilles; Aboudharam, Gérard; Drancourt, Michel

    2016-01-01

    Methanogens are acknowledged archaeal members of modern dental calculus microbiota and dental pathogen complexes. Their repertoire in ancient dental calculus is poorly known. We therefore investigated archaea in one hundred dental calculus specimens collected from individuals recovered from six archaeological sites in France dated from the 14th to 19th centuries AD. Dental calculus was demonstrated by macroscopic and cone-beam observations. In 56 calculus specimens free of PCR inhibition, PCR sequencing identified Candidatus Methanobrevibacter sp. N13 in 44.6%, Methanobrevibacter oralis in 19.6%, a new Methanomassiliicoccus luminyensis-like methanogen in 12.5%, a Candidatus Nitrososphaera evergladensis-like in one and Methanoculleus bourgensis in one specimen, respectively. One Candidatus Methanobrevibacter sp. N13 dental calculus was further documented by fluorescent in situ hybridization. The prevalence of dental calculus M. oralis was significantly lower in past populations than in modern populations (P = 0.03, Chi-square test). This investigation revealed a previously unknown repertoire of archaea found in the oral cavity of past French populations as reflected in preserved dental calculus. PMID:27166431

  13. Recovery of palladium(II) by methanogenic granular sludge.

    PubMed

    Pat-Espadas, Aurora M; Field, James A; Otero-Gonzalez, Lila; Razo-Flores, Elías; Cervantes, Francisco J; Sierra-Alvarez, Reyes

    2016-02-01

    This is the first report that demonstrates the ability of anaerobic methanogenic granular sludge to reduce Pd(II) to Pd(0). Different electron donors were evaluated for their effectiveness in promoting Pd reduction. Formate and H2 fostered both chemically and biologically mediated Pd reduction. Ethanol only promoted the reduction of Pd(II) under biotic conditions and the reduction was likely mediated by H2 released from ethanol fermentation. No reduction was observed in biotic or abiotic assays with all other substrates tested (acetate, lactate and pyruvate) although a large fraction of the total Pd was removed from the liquid medium likely due to biosorption. Pd(II) displayed severe inhibition towards acetoclastic and hydrogenotrophic methanogens, as indicated by 50% inhibiting concentrations as low as 0.96 and 2.7 mg/L, respectively. The results obtained indicate the potential of utilizing anaerobic granular sludge bioreactor technology as a practical and promising option for Pd(II) reduction and recovery offering advantages over pure cultures. PMID:26408982

  14. Absolute Quantification of Individual Biomass Concentrations in a Methanogenic Coculture

    PubMed Central

    2014-01-01

    Identification of individual biomass concentrations is a crucial step towards an improved understanding of anaerobic digestion processes and mixed microbial conversions in general. The knowledge of individual biomass concentrations allows for the calculation of biomass specific conversion rates which form the basis of anaerobic digestion models. Only few attempts addressed the absolute quantification of individual biomass concentrations in methanogenic microbial ecosystems which has so far impaired the calculation of biomass specific conversion rates and thus model validation. This study proposes a quantitative PCR (qPCR) approach for the direct determination of individual biomass concentrations in methanogenic microbial associations by correlating the native qPCR signal (cycle threshold, Ct) to individual biomass concentrations (mg dry matter/L). Unlike existing methods, the proposed approach circumvents error-prone conversion factors that are typically used to convert gene copy numbers or cell concentrations into actual biomass concentrations. The newly developed method was assessed and deemed suitable for the determination of individual biomass concentrations in a defined coculture of Desulfovibrio sp. G11 and Methanospirillum hungatei JF1. The obtained calibration curves showed high accuracy, indicating that the new approach is well suited for any engineering applications where the knowledge of individual biomass concentrations is required. PMID:24949269

  15. Seasonal dynamics of bacterial and archaeal methanogenic communities in flooded rice fields and effect of drainage

    PubMed Central

    Breidenbach, Björn; Conrad, Ralf

    2015-01-01

    We studied the resident (16S rDNA) and the active (16S rRNA) members of soil archaeal and bacterial communities during rice plant development by sampling three growth stages (vegetative, reproductive and maturity) under field conditions. Additionally, the microbial community was investigated in two non-flooded fields (unplanted, cultivated with upland maize) in order to monitor the reaction of the microbial communities to non-flooded, dry conditions. The abundance of Bacteria and Archaea was monitored by quantitative PCR showing an increase in 16S rDNA during reproductive stage and stable 16S rRNA copies throughout the growth season. Community profiling by T-RFLP indicated a relatively stable composition during rice plant growth whereas pyrosequencing revealed minor changes in relative abundance of a few bacterial groups. Comparison of the two non-flooded fields with flooded rice fields showed that the community composition of the Bacteria was slightly different, while that of the Archaea was almost the same. Only the relative abundance of Methanosarcinaceae and Soil Crenarchaeotic Group increased in non-flooded vs. flooded soil. The abundance of bacterial and archaeal 16S rDNA copies was highest in flooded rice fields, followed by non-flooded maize and unplanted fields. However, the abundance of ribosomal RNA (active microbes) was similar indicating maintenance of a high level of ribosomal RNA under the non-flooded conditions, which were unfavorable for anaerobic bacteria and methanogenic archaea. This maintenance possibly serves as preparedness for activity when conditions improve. In summary, the analyses showed that the bacterial and archaeal communities inhabiting Philippine rice field soil were relatively stable over the season but reacted upon change in field management. PMID:25620960

  16. Taxonomic status and ecologic function of methanogenic bacteria isolated from the oral cavity of humans

    SciTech Connect

    Kemp, C.W.

    1985-01-01

    The detection of methane gas in samples of dental plaque and media inoculated with dental plaque was attributed to the presence of methane-producing bacteria in the plaque microbiota. The results of a taxonomic analysis of the 12 methanogenic isolates obtained from human dental plaque, (ABK1-ABK12), placed the organisms in the genus Methanobrevibacter. A DNA-DNA hybridization survey established three distinct genetic groups of oral methanogens based on percent homology values. The groups exhibited less than 32% homology between themselves and less than 17% homology with the three known members of the genus methanobrevibacter. The ecological role of the oral methanogens was established using mixed cultures of selected methanogenic isolates (ABK1, ABK4, ABK6, or ABK7) with oral heterotrophic bacteria. Binary cultures of either Streptococcus mutans, Streptococcus sanguis, Veillonella rodentium, Lactobacillus casei, or Peptostreptococcus anaerobius together with either methanogenic isolates ABK6 or ABK7 were grown to determine the effect of the methanogens on the distribution of carbon end products produced by the heterotrophs. Binary cultures of S. mutans and ABK7 exhibited a 27% decrease in lactic acid formation when compared to pure culture of S. mutans. The decrease in lactic acid production was attributed to the removal of formate by the methanogen, (ABK7), which caused an alteration in the distribution of carbon end products by S. mutans.

  17. “Methanoplasmatales,” Thermoplasmatales-Related Archaea in Termite Guts and Other Environments, Are the Seventh Order of Methanogens

    PubMed Central

    Paul, Kristina; Nonoh, James O.; Mikulski, Lena

    2012-01-01

    The Euryarchaeota comprise both methanogenic and nonmethanogenic orders and many lineages of uncultivated archaea with unknown properties. One of these deep-branching lineages, distantly related to the Thermoplasmatales, has been discovered in various environments, including marine habitats, soil, and also the intestinal tracts of termites and mammals. By comparative phylogenetic analysis, we connected this lineage of 16S rRNA genes to a large clade of unknown mcrA gene sequences, a functional marker for methanogenesis, obtained from the same habitats. The identical topologies of 16S rRNA and mcrA gene trees and the perfect congruence of all branches, including several novel groups that we obtained from the guts of termites and cockroaches, strongly suggested that they stem from the same microorganisms. This was further corroborated by two highly enriched cultures of closely related methanogens from the guts of a higher termite (Cubitermes ugandensis) and a millipede (Anadenobolus sp.), which represented one of the arthropod-specific clusters in the respective trees. Numerous other pairs of habitat-specific sequence clusters were obtained from the guts of other termites and cockroaches but were also found in previously published data sets from the intestinal tracts of mammals (e.g., rumen cluster C) and other environments. Together with the recently described Methanomassiliicoccus luminyensis isolated from human feces, which falls into rice cluster III, the results of our study strongly support the idea that the entire clade of “uncultured Thermoplasmatales” in fact represents the seventh order of methanogenic archaea, for which the provisional name “Methanoplasmatales” is proposed. PMID:23001661

  18. Cultivation of parasites

    PubMed Central

    Ahmed, Nishat Hussain

    2014-01-01

    Parasite cultivation techniques constitute a substantial segment of present-day study of parasites, especially of protozoa. Success in establishing in vitro and in vivo culture of parasites not only allows their physiology, behavior and metabolism to be studied dynamically, but also allows the nature of the antigenic molecules in the excretory and secretory products to be vigorously pursued and analyzed. The complex life-cycles of various parasites having different stages and host species requirements, particularly in the case of parasitic helminths, often make parasite cultivation an uphill assignment. Culturing of parasites depends on the combined expertise of all types of microbiological cultures. Different parasites require different cultivation conditions such as nutrients, temperature and even incubation conditions. Cultivation is an important method for diagnosis of many clinically important parasites, for example, Entamoeba histolytica, Trichomonas vaginalis, Leishmania spp., Strongyloides stercoralis and free-living amoebae. Many commercial systems like InPouch TV for T. vaginalis, microaerophilous stationary phase culture for Babesia bovis and Harada-Mori culture technique for larval-stage nematodes have been developed for the rapid diagnosis of the parasitic infections. Cultivation also has immense utility in the production of vaccines, testing vaccine efficacy, and antigen - production for obtaining serological reagents, detection of drug-resistance, screening of potential therapeutic agents and conducting epidemiological studies. Though in vitro cultivation techniques are used more often compared with in vivo techniques, the in vivo techniques are sometimes used for diagnosing some parasitic infections such as trypanosomiasis and toxoplasmosis. Parasite cultivation continues to be a challenging diagnostic option. This review provides an overview of intricacies of parasitic culture and update on popular methods used for cultivating parasites. PMID

  19. Selenocysteine, Pyrrolysine, and the Unique Energy Metabolism of Methanogenic Archaea

    DOE PAGESBeta

    Rother, Michael; Krzycki, Joseph A.

    2010-01-01

    Methanogenic archaea are a group of strictly anaerobic microorganisms characterized by their strict dependence on the process of methanogenesis for energy conservation. Among the archaea, they are also the only known group synthesizing proteins containing selenocysteine or pyrrolysine. All but one of the known archaeal pyrrolysine-containing and all but two of the confirmed archaeal selenocysteine-containing protein are involved in methanogenesis. Synthesis of these proteins proceeds through suppression of translational stop codons but otherwise the two systems are fundamentally different. This paper highlights these differences and summarizes the recent developments in selenocysteine- and pyrrolysine-related research on archaea and aims to putmore » this knowledge into the context of their unique energy metabolism.« less

  20. Conversion of indole to oxindole under methanogenic conditions

    SciTech Connect

    Berry, D.F.; Madsen, E.L.; Bollag, J.M.

    1987-01-01

    Aromatic N-heterocyclic compounds are often present in aqueous effluents associated with coal mining and processing operations. The environmental fate of these chemicals is of great concern because they are toxic and may contaminate both surface water and groundwater. Previous investigations of microbial metabolism of aromatic chemicals under aerobic and anaerobic conditions suggest that microorganisms may play a key role in determining the fate of this class of compounds. When indole was incubated under methanogenic conditions with an inoculum of sewage sludge, the chemical was metabolized within 10 days and temporary formation of an intermediate was observed. The metabolite was isolated by thin-layer chromatography and determined to be 1,3-dihydro-2H-indol-2-one (oxindole) by UV spectroscopy (lambda/sub max/, 247 nm) and mass spectrometry (m/z, 133). The methane produced (net amount) indicated nearly complete mineralization of indole.

  1. Methanogenic burst in the end-Permian carbon cycle

    PubMed Central

    Rothman, Daniel H.; Fournier, Gregory P.; French, Katherine L.; Alm, Eric J.; Boyle, Edward A.; Cao, Changqun; Summons, Roger E.

    2014-01-01

    The end-Permian extinction is associated with a mysterious disruption to Earth’s carbon cycle. Here we identify causal mechanisms via three observations. First, we show that geochemical signals indicate superexponential growth of the marine inorganic carbon reservoir, coincident with the extinction and consistent with the expansion of a new microbial metabolic pathway. Second, we show that the efficient acetoclastic pathway in Methanosarcina emerged at a time statistically indistinguishable from the extinction. Finally, we show that nickel concentrations in South China sediments increased sharply at the extinction, probably as a consequence of massive Siberian volcanism, enabling a methanogenic expansion by removal of nickel limitation. Collectively, these results are consistent with the instigation of Earth’s greatest mass extinction by a specific microbial innovation. PMID:24706773

  2. Osmoregulation in methanogens. Progress report, May 15, 1991--January 15, 1993

    SciTech Connect

    Roberts, M.F.

    1993-01-01

    Our major goal of our work has been to develop and use NMR techniques to study how methanogenic archaebacteria deal with osmotic stress with the hope of providing insights into increasing the salt tolerance of other cells. The project has three main sections: (i) in vivo studies of methanogens; (ii) use of {sup l3}C- and {sup l5}N- labeled potential precursors and in vitro analyses of specific label uptake for elucidation of osmolyte dynamics and biosynthetic pathways of osmolytes in these organisms, and isolation of key biosynthetic enzymes; and (iii) collaborative studies on identification of organic solutes in other methanogens.

  3. Quantification of methanogenic groups in anaerobic biological reactors by oligonucleotide probe hybridization.

    PubMed Central

    Raskin, L; Poulsen, L K; Noguera, D R; Rittmann, B E; Stahl, D A

    1994-01-01

    The microbial community structure of anaerobic biological reactors was evaluated by using oligonucleotide probes complementary to conserved tracts of the 16S rRNAs of phylogenetically defined groups of methanogens. Phylogenetically defined groups of methanogens were quantified and visualized, respectively, by hybridization of 32P- and fluorescent-dye-labeled probes to the 16S rRNAs from samples taken from laboratory acetate-fed chemostats, laboratory municipal solid waste digestors, and full-scale sewage sludge digestors. Methanosarcina species, members of the order Methanobacteriales, and Methanosaeta species were the most abundant methanogens present in the chemostats, the solid-waste digestors, and the sewage sludge digestors, respectively. PMID:7517129

  4. Progression of methanogenic degradation of crude oil in the subsurface

    USGS Publications Warehouse

    Bekins, B.A.; Hostettler, F.D.; Herkelrath, W.N.; Delin, G.N.; Warren, E.; Essaid, H.I.

    2005-01-01

    Our results show that subsurface crude-oil degradation rates at a long-term research site were strongly influenced by small-scale variations in hydrologic conditions. The site is a shallow glacial outwash aquifer located near Bemidji in northern Minnesota that became contaminated when oil spilled from a broken pipeline in August 1979. In the study area, separate-phase oil forms a subsurface oil body extending from land surface to about 1 m (3.3 ft) below the 6-8-m (20-26 ft)-deep water table. Oil saturation in the sediments ranges from 10-20% in the vadose zone to 30-70% near the water table. At depths below 2 m (6.6 ft), degradation of the separate-phase crude oil occurs under methanogenic conditions. The sequence of methanogenic alkane degradation depletes the longer chain n-alkanes before the shorter chain n-alkanes, which is opposite to the better known aerobic sequence. The rates of degradation vary significantly with location in the subsurface. Oil-coated soils within 1.5 m (5 ft) of land surface have experienced little degradation where soil water saturation is less than 20%. Oil located 2-8 m (6.6-26 ft) below land surface in areas of higher recharge has been substantially degraded. The best explanation for the association between recharge and enhanced degradation seems to be increased downward transport of microbial growth nutrients to the oil body. This is supported by observations of greater microbial numbers at higher elevations in the oil body and significant decreases with depth in nutrient concentrations, especially phosphorus. Our results suggest that environmental effects may cause widely diverging degradation rates in the same spill, calling into question dating methods based on degradation state. Copyright ?? 2005. The American Association of Petroleum Geologists/Division of Environmental Geosciences. All rights reserved.

  5. A Methodological Investigation of Cultivation.

    ERIC Educational Resources Information Center

    Rubin, Alan M.; And Others

    Cultivation theory states that television engenders negative emotions in heavy viewers. Noting that cultivation methodology contains an apparent response bias, a study examined relationships between television exposure and positive restatements of cultivation concepts and tested a more instrumental media uses and effects model. Cultivation was…

  6. Assessing the Ecophysiology of Methanogens in the Context of Recent Astrobiological and Planetological Studies

    NASA Astrophysics Data System (ADS)

    Taubner, Ruth-Sophie; Schleper, Christa; Firneis, Maria G.; Rittman, Simon K.-M. R.

    2015-12-01

    Among all known microbes capable of thriving under extreme and, therefore, potentially extraterrestrial environmental conditions, methanogens from the domain Archaea are intriguing organisms. This is due to their broad metabolic versatility, enormous diversity, and ability to grow under extreme environmental conditions. Several studies revealed that growth conditions of methanogens are compatible with environmental conditions on extraterrestrial bodies throughout the Solar System. Hence, life in the Solar System might not be limited to the classical habitable zone. In this contribution we assess the main ecophysiological characteristics of methanogens and compare these to the environmental conditions of putative habitats in the Solar System, in particular Mars and icy moons. Eventually, we give an outlook on the feasibility and the necessity of future astrobiological studies concerning methanogens.

  7. Transformations of 1- and 2-carbon halogenated aliphatic organic compounds under methanogenic conditions.

    PubMed Central

    Bouwer, E J; McCarty, P L

    1983-01-01

    Several 1- and 2-carbon halogenated aliphatic organic compounds present at low concentrations (less than 100 micrograms/liter) were degraded under methanogenic conditions in batch bacterial cultures and in a continuous-flow methanogenic fixed-film laboratory-scale column. Greater than 90% degradation was observed within a 2-day detention time under continuous-flow methanogenic conditions with acetate as a primary substrate. Carbon-14 measurements indicated that chloroform, carbon tetrachloride, and 1,2-dichloroethane were almost completely oxidized to carbon dioxide, confirming removal by biooxidation. The initial step in the transformations of tetrachloroethylene and 1,1,2,2-tetrachloroethane to nonchlorinated end products appeared to be reductive dechlorination to trichloroethylene and 1,1,2-trichloroethane, respectively. Transformations of the brominated aliphatic compounds appear to be the result of both biological and chemical processes. The data suggest that transformations of halogenated aliphatic compounds can occur under methanogenic conditions in the environment. PMID:6859849

  8. Assessing the Ecophysiology of Methanogens in the Context of Recent Astrobiological and Planetological Studies.

    PubMed

    Taubner, Ruth-Sophie; Schleper, Christa; Firneis, Maria G; Rittmann, Simon K-M R

    2015-01-01

    Among all known microbes capable of thriving under extreme and, therefore, potentially extraterrestrial environmental conditions, methanogens from the domain Archaea are intriguing organisms. This is due to their broad metabolic versatility, enormous diversity, and ability to grow under extreme environmental conditions. Several studies revealed that growth conditions of methanogens are compatible with environmental conditions on extraterrestrial bodies throughout the Solar System. Hence, life in the Solar System might not be limited to the classical habitable zone. In this contribution we assess the main ecophysiological characteristics of methanogens and compare these to the environmental conditions of putative habitats in the Solar System, in particular Mars and icy moons. Eventually, we give an outlook on the feasibility and the necessity of future astrobiological studies concerning methanogens. PMID:26703739

  9. SEQUENTIAL REDUCTIVE DEHALOGATION OF CHLOROANILINES BY MICROORGANISMS FROM A METHANOGENIC AQUIFER

    EPA Science Inventory

    Chloroaniline-based compounds are widely used chem- icals and important contaminants of aquatic and terrestrial environments. We have found that chloroanilines can be biologically dehalogenated in polluted aquifers when methanogenic, but not sulfate-reducing conditions prevail. T...

  10. SEQUENTIAL REDUCTIVE DEHALOGENATION OF CHLORANILINES BY MICROORGANISMS FROM A METHANOGENIC AQUIFER

    EPA Science Inventory

    Chloroaniline-based compounds are widely used chemicals and important contaminants of aquatic and terrestrial environments. We have found that chloroanilines can be bio-logically dehalogenated in polluted aquifers when methanogenic, but not sulfate-reducing conditions prevail. Th...

  11. BIOTRANSFORMATIONS OF SELECTED ALKYLBENZENES AND HALOGENATED ALIPHATIC HYDROCARBONS IN METHANOGENIC AQUIFER MATERIAL: A MICROCOSM STUDY

    EPA Science Inventory

    Leachates from municipal landfills commonly contain a variety of organic contaminants of industrial origin. The behavior of these compounds in anaerobic, and particularly in methanogenic, subsurface materials is poorly understood. The behavior of benzene, toluene, ethylbenzene, o...

  12. Assessing the Ecophysiology of Methanogens in the Context of Recent Astrobiological and Planetological Studies

    PubMed Central

    Taubner, Ruth-Sophie; Schleper, Christa; Firneis, Maria G.; Rittmann, Simon K.-M. R.

    2015-01-01

    Among all known microbes capable of thriving under extreme and, therefore, potentially extraterrestrial environmental conditions, methanogens from the domain Archaea are intriguing organisms. This is due to their broad metabolic versatility, enormous diversity, and ability to grow under extreme environmental conditions. Several studies revealed that growth conditions of methanogens are compatible with environmental conditions on extraterrestrial bodies throughout the Solar System. Hence, life in the Solar System might not be limited to the classical habitable zone. In this contribution we assess the main ecophysiological characteristics of methanogens and compare these to the environmental conditions of putative habitats in the Solar System, in particular Mars and icy moons. Eventually, we give an outlook on the feasibility and the necessity of future astrobiological studies concerning methanogens. PMID:26703739

  13. Colonization of rice roots with methanogenic archaea controls photosynthesis-derived methane emission.

    PubMed

    Pump, Judith; Pratscher, Jennifer; Conrad, Ralf

    2015-07-01

    The methane emitted from rice fields originates to a large part (up to 60%) from plant photosynthesis and is formed on the rice roots by methanogenic archaea. To investigate to which extent root colonization controls methane (CH4 ) emission, we pulse-labeled rice microcosms with (13) CO2 to determine the rates of (13) CH4 emission exclusively derived from photosynthates. We also measured emission of total CH4 ((12+13) CH4 ), which was largely produced in the soil. The total abundances of archaea and methanogens on the roots and in the soil were analysed by quantitative polymerase chain reaction of the archaeal 16S rRNA gene and the mcrA gene coding for a subunit of the methyl coenzyme M reductase respectively. The composition of archaeal and methanogenic communities was determined with terminal restriction fragment length polymorphism (T-RFLP). During the vegetative growth stages, emission rates of (13) CH4 linearly increased with the abundance of methanogenic archaea on the roots and then decreased during the last plant growth stage. Rates of (13) CH4 emission and the abundance of methanogenic archaea were lower when the rice was grown in quartz-vermiculite with only 10% rice soil. Rates of total CH4 emission were not systematically related to the abundance of methanogenic archaea in soil plus roots. The composition of the archaeal communities was similar under all conditions; however, the analysis of mcrA genes indicated that the methanogens differed between the soil and root. Our results support the hypothesis that rates of photosynthesis-driven CH4 emission are limited by the abundance of methanogens on the roots. PMID:25367104

  14. Mesophilic fermentation of renewable biomass: does hydraulic retention time regulate methanogen diversity?

    PubMed

    Krakat, Niclas; Schmidt, Stefan; Scherer, Paul

    2010-09-01

    The present long-term study (about 1,100 days) monitored the diversity of methanogens during the mesophilic, anaerobic digestion of beet silage. Six fermentor samples were analyzed by ribosomal RNA gene restriction analysis, fluorescence in situ hybridization, and fluorescence microscopy. Hydrogenotrophic methanogens dominated within the population in all samples analyzed. Multidimensional scaling revealed that a rapid decrease in hydraulic retention time resulted in increased species richness, which in turn led to slightly higher CH(4) yields. PMID:20675458

  15. A T-RFLP database for the rapid profiling of methanogenic communities in anaerobic digesters.

    PubMed

    Bühligen, Franziska; Lucas, Rico; Nikolausz, Marcell; Kleinsteuber, Sabine

    2016-06-01

    We present a simple protocol for the cost- and time-efficient profiling of methanogens based on T-RFLP fingerprinting of mcrA amplicons. Sequence data were compiled from mesophilic lab-scale and full-scale biogas reactors operated under various conditions and fed with various substrates. The database facilitates the rapid identification of methanogens, thus reducing the need of cloning and sequencing. PMID:27046270

  16. Methanogenic archaea diversity in hyporheic sediments of a small lowland stream.

    PubMed

    Brablcová, Lenka; Buriánková, Iva; Badurová, Pavlína; Chaudhary, Prem Prashant; Rulík, Martin

    2015-04-01

    Abundance and diversity of methanogenic archaea were studied at five localities along a longitudinal profile of a Sitka stream (Czech Republic). Samples of hyporheic sediments were collected from two sediment depths (0-25 cm and 25-50 cm) by freeze-core method. Methanogen community was analyzed by fluorescence in situ hybridization (FISH), denaturing gradient gel electrophoresis (DGGE) and sequencing method. The proportion of methanogens to the DAPI-stained cells varied among all localities and depths with an average value 2.08 × 10(5) per g of dry sediment with the range from 0.37 to 4.96 × 10(5) cells per g of dry sediment. A total of 73 bands were detected at 19 different positions on the DGGE gel and the highest methanogen diversity was found at the downstream located sites. There was no relationship between methanogen diversity and sediment depth. Cluster analysis of DGGE image showed three main clusters consisting of localities that differed in the number and similarity of the DGGE bands. Sequencing analysis of representative DGGE bands revealed phylotypes affiliated with members belonging to the orders Methanosarcinales, Methanomicrobiales and Methanocellales. The knowledge about occurrence and diversity of methanogenic archaea in freshwater ecosystems are essential for methane dynamics in river sediments and can contribute to the understanding of global warming process. PMID:25460192

  17. Performance of methanogenic reactors in temperature phased two-stage anaerobic digestion of swine wastewater.

    PubMed

    Kim, Woong; Shin, Seung Gu; Cho, Kyungjin; Lee, Changsoo; Hwang, Seokhwan

    2012-12-01

    The present study investigated the shifts in the chemical profiles of a two-phase anaerobic digestion system in methanogenic and acidogenic reactors for the treatment of swine wastewater. Acidogenic and methanogenic digesters were used with overall HRTs ranging from 27 to 6 d. In the optimized thermophilic/acidogenic phase throughout the entire experimental period, VS was reduced by 13.8% (1.6%); however, COD hardly decreased because of the thermophilic hydrolysis of organic materials, such as carbohydrates, proteins, and lipids, without any significant consumption of volatile fatty acids. In the methanogenic/mesophilic phase, COD was reduced by 65.8 (1.1)% compared to a 47.4 (2.9)% reduction in VS reduction efficiency with the gradual increase in methane production during a methanogenic HRT between 25 and 10 d. A high protein degradation rate was observed in the optimized acidogenic phase, which is assumed to be due to the low content of carbohydrates in raw swine wastewater as well as the readily thermophilic hydrolysis of proteins. Two-phase systems of anaerobic digestion consisting of optimized thermophilic and mesophilic methanogenic digesters showed a stable performance with respect to VS reduction efficiency with OLRs less than 3 g VS/L·d, in other words, more than 10 days of methanogenic HRT in this study. PMID:23041140

  18. Methanogenic archaea in marcellus shale: a possible mechanism for enhanced gas recovery in unconventional shale resources.

    PubMed

    Tucker, Yael Tarlovsky; Kotcon, James; Mroz, Thomas

    2015-06-01

    Marcellus Shale occurs at depths of 1.5-2.5 km (5000 to 8000 feet) where most geologists generally assume that thermogenic processes are the only source of natural gas. However, methanogens in produced fluids and isotopic signatures of biogenic methane in this deep shale have recently been discovered. This study explores whether those methanogens are indigenous to the shale or are introduced during drilling and hydraulic fracturing. DNA was extracted from Marcellus Shale core samples, preinjected fluids, and produced fluids and was analyzed using Miseq sequencing of 16s rRNA genes. Methanogens present in shale cores were similar to methanogens in produced fluids. No methanogens were detected in injected fluids, suggesting that this is an unlikely source and that they may be native to the shale itself. Bench-top methane production tests of shale core and produced fluids suggest that these organisms are alive and active under simulated reservoir conditions. Growth conditions designed to simulate the hydrofracture processes indicated somewhat increased methane production; however, fluids alone produced relatively little methane. Together, these results suggest that some biogenic methane may be produced in these wells and that hydrofracture fluids currently used to stimulate gas recovery could stimulate methanogens and their rate of producing methane. PMID:25924080

  19. Reduction of hexavalent chromium by the thermophilic methanogen Methanothermobacter thermautotrophicus

    NASA Astrophysics Data System (ADS)

    Singh, Rajesh; Dong, Hailiang; Liu, Deng; Zhao, Linduo; Marts, Amy R.; Farquhar, Erik; Tierney, David L.; Almquist, Catherine B.; Briggs, Brandon R.

    2015-01-01

    Despite significant progress on iron reduction by thermophilic microorganisms, studies on their ability to reduce toxic metals are still limited, despite their common co-existence in high temperature environments (up to 70 °C). In this study, Methanothermobacter thermautotrophicus, an obligate thermophilic methanogen, was used to reduce hexavalent chromium. Experiments were conducted in a growth medium with H2/CO2 as substrate with various Cr6+ concentrations (0.2, 0.4, 1, 3, and 5 mM) in the form of potassium dichromate (K2Cr2O7). Time-course measurements of aqueous Cr6+ concentrations using 1,5-diphenylcarbazide colorimetric method showed complete reduction of the 0.2 and 0.4 mM Cr6+ solutions by this methanogen. However, much lower reduction extents of 43.6%, 13.0%, and 3.7% were observed at higher Cr6+ concentrations of 1, 3 and 5 mM, respectively. These lower extents of bioreduction suggest a toxic effect of aqueous Cr6+ to cells at this concentration range. At these higher Cr6+ concentrations, methanogenesis was inhibited and cell growth was impaired as evidenced by decreased total cellular protein production and live/dead cell ratio. Likewise, Cr6+ bioreduction rates decreased with increased initial concentrations of Cr6+ from 13.3 to 1.9 μM h-1. X-ray absorption near-edge structure (XANES) spectroscopy revealed a progressive reduction of soluble Cr6+ to insoluble Cr3+ precipitates, which was confirmed as amorphous chromium hydroxide by selected area electron diffraction pattern. However, a small fraction of reduced Cr occurred as aqueous Cr3+. Scanning and transmission electron microscope observations of M. thermautotrophicus cells after Cr6+ exposure suggest both extra- and intracellular chromium reduction mechanisms. Results of this study demonstrate the ability of M. thermautotrophicus cells to reduce toxic Cr6+ to less toxic Cr3+ and its potential application in metal bioremediation, especially at high temperature subsurface radioactive waste disposal

  20. Methanogenic Oil Degradation in the Dagang Oil Field

    NASA Astrophysics Data System (ADS)

    Jiménez, Núria; Cai, Minmin; Straaten, Nontje; Yao, Jun; Richnow, Hans Hermann; Krüger, Martin

    2014-05-01

    Anaerobic biodegradation is one of the main in situ oil transformation processes in subsurface oil reservoirs. Recent studies have provided evidence of biodegradation of residual oil constituents under methanogenic conditions. Methane, like other biogenic gases, may contribute to reduce the viscosity of oil and enhance its flow characteristics (making it more available) but it can also be used as a energy source. So the aim of the present study was to provide reliable information on in situ biotransformation of oil under methanogenic conditions, and to assess the feasibility of implementing a MEOR strategy at this site. For this reason, chemical and isotopic analyses of injection and production fluids of the Dagang oil field (Hebei province, China) were performed. Microbial abundances were assessed by qPCR, and clone libraries were performed to study the diversity. In addition, microcosms with either oil or 13C-labelled hydrocarbons were inoculated with injection or production waters to characterize microbial processes in vitro. Geochemical and isotopic data were consistent with in situ biogenic methane production linked to aliphatic and aromatic hydrocarbon degradation: GC-MS profiles of petroleum samples were nearly devoid of n-alkanes, linear alkylbenzenes, and alkyltoluenes, and light PAH, confirming that Dagang oil is mostly highly weathered. In addition, carbon and hydrogen isotopic signatures of methane (δ13CCH4 and δDCH4, respectively), and the bulk isotopic discrimination (Δδ13C) between methane and CO2 (between 32 and 65 ) were in accordance with previously reported values for methane formation during hydrocarbon degradation. Furthermore, methane-producing Archaea and hydrocarbon-degrading Bacteria were abundant in produced oil-water samples. On the other hand, our laboratory degradation experiments revealed that autochthonous microbiota are capable of significantly degrade oil within several months, with biodegradation patterns resembling those

  1. Reduction of hexavalent chromium by the thermophilic methanogen Methanothermobacter thermautotrophicus

    DOE PAGESBeta

    Singh, Rajesh; Dong, Hailiang; Liu, Deng; Zhao, Linduo; Marts, Amy R.; Farquhar, Erik; Tierney, David L.; Almquist, Catherine B.; Briggs, Brandon R.

    2014-10-22

    Despite the significant progress on iron reduction by thermophilic microorganisms, studies on their ability to reduce toxic metals are still limited, despite their common co-existence in high temperature environments (up to 70°C). In this study, Methanothermobacter thermautotrophicus, an obligate thermophilic methanogen, was used to reduce hexavalent chromium. Experiments were conducted in a growth medium with H2/CO2 as substrate with various Cr6+ concentrations (0.2, 0.4, 1, 3, and 5 mM) in the form of potassium dichromate (K2Cr2O7). Time-course measurements of aqueous Cr6+ concentrations with the 1, 5-diphenylcarbazide colorimetric method showed complete reduction of the 0.2 and 0.4 mM Cr6+ solutions bymore » this methanogen. However, much lower reduction extents of 43.6%, 13.0%, and 3.7% were observed at higher Cr6+ concentrations of 1, 3 and 5 mM, respectively. These lower extents of bioreduction suggest a toxic effect of aqueous Cr6+ to cells at this concentration range. At these higher Cr6+ concentrations, methanogenesis was inhibited and cell growth was impaired as evidenced by decreased total cellular protein production and live/dead cell ratio. Likewise, Cr6+ bioreduction rates decreased with increased initial concentrations of Cr6+ from 13.3 to1.9 μM h₋1. X-ray absorption near-edge structure (XANES) spectroscopy revealed a progressive reduction of soluble Cr6+ to insoluble Cr3+ precipitates, which was confirmed as amorphous chromium hydroxide by X-ray diffraction and selected area electron diffraction pattern. However, a small fraction of reduced Cr occurred as aqueous Cr3+. Scanning and transmission electron microscope observations of M. thermautotrophicus cells after Cr6+ exposure suggest both extra- and intracellular chromium reduction mechanisms. Results of this study demonstrate the ability of M. thermautotrophicus cells to reduce toxic Cr6+ to less toxic Cr3+ and its potential application in metal bioremediation, especially at high temperature

  2. Reduction of hexavalent chromium by the thermophilic methanogen Methanothermobacter thermautotrophicus

    PubMed Central

    Singh, Rajesh; Dong, Hailiang; Liu, Deng; Zhao, Linduo; Marts, Amy R.; Farquhar, Erik; Tierney, David L.; Almquist, Catherine B.; Briggs, Brandon R.

    2015-01-01

    Despite the significant progress on iron reduction by thermophilic microorganisms, studies on their ability to reduce toxic metals are still limited, despite their common co-existence in high temperature environments (up to 70°C). In this study, Methanothermobacter thermautotrophicus, an obligate thermophilic methanogen, was used to reduce hexavalent chromium. Experiments were conducted in a growth medium with H2/CO2 as substrate with various Cr6+ concentrations (0.2, 0.4, 1, 3, and 5 mM) in the form of potassium dichromate (K2Cr2O7). Time-course measurements of aqueous Cr6+ concentrations with the 1, 5-diphenylcarbazide colorimetric method showed complete reduction of the 0.2 and 0.4 mM Cr6+ solutions by this methanogen. However, much lower reduction extents of 43.6%, 13.0%, and 3.7% were observed at higher Cr6+ concentrations of 1, 3 and 5 mM, respectively. These lower extents of bioreduction suggest a toxic effect of aqueous Cr6+ to cells at this concentration range. At these higher Cr6+ concentrations, methanogenesis was inhibited and cell growth was impaired as evidenced by decreased total cellular protein production and live/dead cell ratio. Likewise, Cr6+ bioreduction rates decreased with increased initial concentrations of Cr6+ from 13.3 to1.9 µM h−1. X-ray absorption near-edge structure (XANES) spectroscopy revealed a progressive reduction of soluble Cr6+ to insoluble Cr3+ precipitates, which was confirmed as amorphous chromium hydroxide by X-ray diffraction and selected area electron diffraction pattern. However, a small fraction of reduced Cr occurred as aqueous Cr3+. Scanning and transmission electron microscope observations of M. thermautotrophicus cells after Cr6+ exposure suggest both extra- and intracellular chromium reduction mechanisms. Results of this study demonstrate the ability of M. thermautotrophicus cells to reduce toxic Cr6+ to less toxic Cr3+ and its potential application in metal bioremediation, especially at high temperature

  3. Reduction of hexavalent chromium by the thermophilic methanogen Methanothermobacter thermautotrophicus

    SciTech Connect

    Singh, Rajesh; Dong, Hailiang; Liu, Deng; Zhao, Linduo; Marts, Amy R.; Farquhar, Erik; Tierney, David L.; Almquist, Catherine B.; Briggs, Brandon R.

    2014-10-22

    Despite the significant progress on iron reduction by thermophilic microorganisms, studies on their ability to reduce toxic metals are still limited, despite their common co-existence in high temperature environments (up to 70°C). In this study, Methanothermobacter thermautotrophicus, an obligate thermophilic methanogen, was used to reduce hexavalent chromium. Experiments were conducted in a growth medium with H2/CO2 as substrate with various Cr6+ concentrations (0.2, 0.4, 1, 3, and 5 mM) in the form of potassium dichromate (K2Cr2O7). Time-course measurements of aqueous Cr6+ concentrations with the 1, 5-diphenylcarbazide colorimetric method showed complete reduction of the 0.2 and 0.4 mM Cr6+ solutions by this methanogen. However, much lower reduction extents of 43.6%, 13.0%, and 3.7% were observed at higher Cr6+ concentrations of 1, 3 and 5 mM, respectively. These lower extents of bioreduction suggest a toxic effect of aqueous Cr6+ to cells at this concentration range. At these higher Cr6+ concentrations, methanogenesis was inhibited and cell growth was impaired as evidenced by decreased total cellular protein production and live/dead cell ratio. Likewise, Cr6+ bioreduction rates decreased with increased initial concentrations of Cr6+ from 13.3 to1.9 μM h₋1. X-ray absorption near-edge structure (XANES) spectroscopy revealed a progressive reduction of soluble Cr6+ to insoluble Cr3+ precipitates, which was confirmed as amorphous chromium hydroxide by X-ray diffraction and selected area electron diffraction pattern. However, a small fraction of reduced Cr occurred as aqueous Cr3+. Scanning and transmission electron microscope observations of M. thermautotrophicus cells after Cr6+ exposure suggest both extra- and intracellular chromium reduction mechanisms. Results of

  4. Relationship between Trophic Status and Methanogenic Pathways in Alaskan Peatlands

    NASA Astrophysics Data System (ADS)

    Zhang, L.; Liu, X.; Sidelinger, W.; Wang, Y.; Hines, M. E.; Langford, L.; Chanton, J.

    2015-12-01

    To improve predictions of naturally emitted CH4 from northern wetlands, it is necessary to further examine the methanogenic pathways in these wetlands. Stable isotope C ratios (δ13C) have been used as a robust tool to distinguish different pathways, but different sources of parent compounds (acetate and CO2) with unique δ13C may add complexity to previously established criteria. Large portions of peatlands accommodate a mixture of different sphagna and sedges. Plant species may look very similar and belong to the same genus but are different morphologically and physiologically. To better understand the relationships between surface vegetation patterns and methanogenic pathways, 26 peatland sites were studied in Fairbanks and Anchorage, Alaska in summers of 2014 and 2015. These sites were ordinated using multiple factor analysis into 3 clusters based on pH, temp, CH4 and volatile fatty acids production rates, δ13C values, and surface vegetation species/pattern. In the low-pH trophic cluster (pH~3.5), non-vascular/vascular plant ratios (NV/V) were ~ 0.87 and dominated by diverse Sphagnum species and specific sedges (Eriophorum vaginatum), and fermentation was the dominant end-point in decomposition with no CH4 detected. Although NV/V is about the same in the intermediate cluster (0.74) (pH~4.5), and Sphagnum squarrosum was largely present, both hydrogenotrophic (HM) and acetoclastic methanogenesis (AM) were very active. Syntrophy was present at certain sites, which may provide CO2 with unique δ13C for CH4 production. At the highest pH trophic cluster examined in this study (pH~5), non-vascular plants were almost not existent and Carex aquatilis dominated. CH4 production rates (mainly HM) were slower than those in the intermediate cluster and the apparent fractionation factor a was lower than in the sites with syntrophy, which warrants further investigation of the position and compound specific δ13C analysis of volatile fatty acids.

  5. Seaweed cultivation for renewable resources

    SciTech Connect

    Bird, K.T.; Benson, P.H.

    1987-01-01

    In the 1970's and 80's, major research and development programs were launched to explore the possibility of using marine biomass as a source of energy. This volume, not only reviews the accomplishments of the aforementioned programs, but also describes how this research relates to seaweed cultivation for other products, such as food, feed, and high value chemicals. Topics covered include the features of marine biomass production, biotechnological manipulations of marine algae, and marine biomass conversion to energy, as well as economics. The chapters synthesize a large number of technical reports, journal articles, symposia and conference proceedings and technology transfer meetings.

  6. Methane production improvement by modulation of solid phase immersion in dry batch anaerobic digestion process: Dynamic of methanogen populations.

    PubMed

    André, L; Ndiaye, M; Pernier, M; Lespinard, O; Pauss, A; Lamy, E; Ribeiro, T

    2016-05-01

    Several 60L dry batch anaerobic digestion (AD) reactors were implemented with or without liquid reserve on cattle manure. The immersed part modulation of cattle manure increased the methane flow of about 13%. The quantitative real time PCR and the optimized DNA extraction were implemented and validated to characterize and quantify the methanogen dynamic in dry batch AD process. Final quantities of methanogens converged toward the same level in several inocula at the end of AD. Methanogen dynamic was shown by dominance of Methanosarcinaceae for acetotrophic methanogens and Methanobacteriales for the hydrogenotrophic methanogens. Overall, methanogens populations were stabilized in liquid phase, except Methanosaetaceae. Solid phase was colonized by Methanomicrobiales and Methanosarcinaceae populations giving a support to biofilm development. The methane increase could be explained by a raise of Methanosarcinaceae population in presence of a total contact between solid and liquid phases. Methanosarcinaceae was a bio-indicator of the methane production. PMID:26897414

  7. Reactor performance of a 750 m(3) anaerobic digestion plant: varied substrate input conditions impacting methanogenic community.

    PubMed

    Wagner, Andreas Otto; Malin, Cornelia; Lins, Philipp; Gstraunthaler, Gudrun; Illmer, Paul

    2014-10-01

    A 750 m(3) anaerobic digester was studied over a half year period including a shift from good reactor performance to a reduced one. Various abiotic parameters like volatile fatty acids (VFA) (formic-, acetic-, propionic-, (iso-)butyric-, (iso-)valeric-, lactic acid), total C, total N, NH4 -N, and total proteins, as well as the organic matter content and dry mass were determined. In addition several process parameters such as temperature, pH, retention time and input of substrate and the concentrations of CH4, H2, CO2 and H2S within the reactor were monitored continuously. The present study aimed at the investigation of the abundance of acetogens and total cell numbers and the microbial methanogenic community as derived from PCR-dHPLC analysis in order to put it into context with the determined abiotic parameters. An influence of substrate quantity on the efficiency of the anaerobic digestion process was found as well as a shift from a hydrogenotrophic in times of good reactor performance towards an acetoclastic dominated methanogenic community in times of reduced reactor performance. After the change in substrate conditions it took the methano-archaeal community about 5-6 weeks to be affected but then changes occurred quickly. PMID:24727280

  8. Cultivating Leaders from Within

    ERIC Educational Resources Information Center

    Burdette, Maggie; Schertzer, Kristen

    2005-01-01

    A major problem faced by school districts in the US is the paucity of applicants for the posts of school principals. A solution adopted by The Capistrano Unified School District (CUSD) in Orange County California was the cultivation of good leaders from within the district through the Teaching Assistant Principal (TAP) program.

  9. Bacterial surface antigens defined by monoclonal antibodies: the methanogens

    SciTech Connect

    Conway de Macario, E.; Macario, A.J.L.; Magarinos, M.C.; Jovell, R.J.; Kandler, O.

    1982-01-01

    The methanogens (MB) are unique microbes of great evolutionary interest with applications in biotechnology-bioengineerings and are important in digestive processes. Their cell-wall composition is distinctively different from that of Eubacteria, e.g. the Methanobacteriaceae possess the peptidoglycan pseudomurein rather than murein. The range of cell-wall compositions among MB and their evolutionary and functional significance is not well known. The authors undertook a systematic study of the MB's surface structure using monoclonal antibodies through the following steps: (1) generation of hybridomas that produce antibody to several MB from 3 of their 4 families; (2) development of immunoenzymatic assays for MB's antigens and antibodies; (3) determination of the fine specificity of monoclonal antibodies by inhibition-blocking tests using cell-wall extracts and compounds of known structure; thus a set of monoclonal probes of predetermined specificity was assembled; and (4) resolution of surface determinants of MB representative of the Methanobacteriaceae using the monoclonal probes. Specific markers of MB strains were characterized. Two epitopes were identified within the pseudomurein molecule.

  10. Reductive dechlorination of chlorophenols by a pentachlorophenol- acclimated methanogenic consortium.

    PubMed Central

    Nicholson, D K; Woods, S L; Istok, J D; Peek, D C

    1992-01-01

    Anaerobic digester sludge fed 5,300 mg of acetate per liter, 3.4 microM pentachlorophenol, and nutrients for 10 days biotransformed pentachlorophenol by sequential ortho dechlorinations to produce 2,3,4,5-tetrachlorophenol and 3,4,5-trichlorophenol. Upon acclimation to 3.4 microM pentachlorophenol for 6 months, the methanogenic consortium removed chlorines from the ortho, meta, and para positions of pentachlorophenol and its reductive dechlorination products. Pentachlorophenol was degraded to produce 2,3,4,5-tetrachlorophenol, 2,3,4,6-tetrachlorophenol, and 2,3,5,6-tetrachlorophenol. Dechlorination of 2,3,4,5-tetrachlorophenol produced 3,4,5-trichlorophenol, which was subsequently degraded to produce 3,4-dichlorophenol and 3,5-dichlorophenol. 2,3,4,6-Tetrachlorophenol was dechlorinated at the ortho and meta positions to produce 2,4,6-trichlorophenol and 2,4,5-trichlorophenol. 2,3,5,6-Tetrachlorophenol yielded 2,3,5-trichlorophenol, followed by production of 3,5-dichlorophenol. 2,4,6-Trichlorophenol was degraded to form 2,4-dichlorophenol, and 2,4,5-trichlorophenol was dechlorinated at two positions to form 2,4-dichlorophenol and 3,4-dichlorophenol. Of the three dichlorophenols produced (2,4-dichlorophenol, 3,4-dichlorophenol, and 3,5-dichlorophenol), only 2,4-dichlorophenol was degraded significantly within 3 weeks, to produce 4-chlorophenol. PMID:1637165

  11. Interaction of acetogens and methanogens in anaerobic freshwater sediments.

    PubMed

    Jones, J G; Simon, B M

    1985-04-01

    Anaerobic decomposition processes in the profundal sediments of Blelham Tarn (English Lake District) are often limited during late summer by the input of organic carbon. The concentration of acetate in the interstitial water fell from about 100 microM (immediately after sedimentation of the spring diatom bloom) to a relatively constant value of about 20 microM in late summer, during which acetate utilization appeared to be balanced by production. Addition of chloroform and molybdate caused an accumulation of cold acetate in large sediment cores and of [14C]acetate in small cores to which [14C]bicarbonate had been added. In both cases chloroform caused the greater accumulation, implying that acetoclastic methanogens were the more active consumers. The conversion of 14CO2 to [14C]acetate was inversely related, with depth, to its conversion to 14CH4. Methanogenesis from CO2 decreased during late summer, whereas acetogenesis and acetoclastic methanogenesis increased over the same time period. The production of acetate from CO2 was generally equivalent to less than 10% of the acetate carbon utilized but could be as high as 25% of that value. Hydrogen consumption by acetogens could be as high as 50% of that utilized in methanogenesis. The role of acetogenic bacteria in anaerobic processes may therefore be of greater significance in lakes such as Blelham Tarn than in more eutrophic systems. PMID:4004224

  12. Continuous culture of Methanococcus jannaschii, an extremely thermophilic methanogen

    SciTech Connect

    Jiahuey Tsao; Kaneshiro, S.M.; Shusan Yu; Clark, D.S. . Dept. of Chemical Engineering)

    1994-02-05

    Methanococcus jannaschii, an extremely thermophilic methanogen isolated from a deep-sea hydrothermal vent was grown at 80 C in continuous culture on a mineral salts medium gassed with H[sub 2] and CO[sub 2] at three different flow rates. The maximum specific growth rate was 0.56 h[sup [minus]1], and the maximum specific methane productivity was 0.32 (mol g[sup [minus]1] h[sup [minus]1]). Uncoupling of growth and methane production was evidenced by an increase in the non-growth-associated rate of methane formation, [Beta], with increasing gaseous input. The specific hydrogenase activity exhibited growth-associated behavior at low growth rates, but showed no dependence on growth at higher growth rates. The growth dependence of hydrogenase activity is consistent with the pressure dependence of hydrogenase activity measured in previous experiments. In contrast, the specific protease activity was independent of the growth rate over the entire range of dilution rates studied.

  13. Methanogenic archaea and sulfate reducing bacteria co-cultured on acetate: teamwork or coexistence?

    PubMed Central

    Ozuolmez, Derya; Na, Hyunsoo; Lever, Mark A.; Kjeldsen, Kasper U.; Jørgensen, Bo B.; Plugge, Caroline M.

    2015-01-01

    Acetate is a major product of fermentation processes and an important substrate for sulfate reducing bacteria and methanogenic archaea. Most studies on acetate catabolism by sulfate reducers and methanogens have used pure cultures. Less is known about acetate conversion by mixed pure cultures and the interactions between both groups. We tested interspecies hydrogen transfer and coexistence between marine methanogens and sulfate reducers using mixed pure cultures of two types of microorganisms. First, Desulfovibrio vulgaris subsp. vulgaris (DSM 1744), a hydrogenotrophic sulfate reducer, was cocultured together with the obligate aceticlastic methanogen Methanosaeta concilii using acetate as carbon and energy source. Next, Methanococcus maripaludis S2, an obligate H2- and formate-utilizing methanogen, was used as a partner organism to M. concilii in the presence of acetate. Finally, we performed a coexistence experiment between M. concilii and an acetotrophic sulfate reducer Desulfobacter latus AcSR2. Our results showed that D. vulgaris was able to reduce sulfate and grow from hydrogen leaked by M. concilii. In the other coculture, M. maripaludis was sustained by hydrogen leaked by M. concilii as revealed by qPCR. The growth of the two aceticlastic microbes indicated co-existence rather than competition. Altogether, our results indicate that H2 leaking from M. concilii could be used by efficient H2-scavengers. This metabolic trait, revealed from coculture studies, brings new insight to the metabolic flexibility of methanogens and sulfate reducers residing in marine environments in response to changing environmental conditions and community compositions. Using dedicated physiological studies we were able to unravel the occurrence of less obvious interactions between marine methanogens and sulfate-reducing bacteria. PMID:26074892

  14. Substrate sources regulate spatial variation of metabolically active methanogens from two contrasting freshwater wetlands.

    PubMed

    Lin, Yongxin; Liu, Deyan; Ding, Weixin; Kang, Hojeong; Freeman, Chris; Yuan, Junji; Xiang, Jian

    2015-12-01

    There is ample evidence that methane (CH4) emissions from natural wetlands exhibit large spatial variations at a field scale. However, little is known about the metabolically active methanogens mediating these differences. We explored the spatial patterns in active methanogens of summer inundated Calamagrostis angustifolia marsh with low CH4 emissions and permanently inundated Carex lasiocarpa marsh with high CH4 emissions in Sanjiang Plain, China. In C. angustifolia marsh, the addition of (13)C-acetate significantly increased the CH4 production rate, and Methanosarcinaceae methanogens were found to participate in the consumption of acetate. In C. lasiocarpa marsh, there was no apparent increase in the CH4 production rate and no methanogen species were labeled with (13)C. When (13)CO2-H2 was added, however, CH4 production was found to be due to Fen Cluster (Methanomicrobiales) in C. angustifolia marsh and Methanobacterium Cluster B (Methanobacteriaceae) together with Fen Cluster in C. lasiocarpa marsh. These results suggested that CH4 was produced primarily by hydrogenotrophic methanogens using substrates mainly derived from plant litter in C. lasiocarpa marsh and by both hydrogenotrophic and acetoclastic methanogens using substrates mainly derived from root exudate in C. angustifolia marsh. The significantly lower CH4 emissions measured in situ in C. angustifolia marsh was primarily due to a deficiency of substrates compared to C. lasiocarpa marsh. Therefore, we speculate that the substrate source regulates both the type of active methanogens and the CH4 production pathway and consequently contributes to the spatial variations in CH4 productions observed in these freshwater marshes. PMID:26286511

  15. The quantitative significance of Syntrophaceae and syntrophic partnerships in methanogenic degradation of crude oil alkanes

    PubMed Central

    Gray, N D; Sherry, A; Grant, R J; Rowan, A K; Hubert, C R J; Callbeck, C M; Aitken, C M; Jones, D M; Adams, J J; Larter, S R; Head, I M

    2011-01-01

    Libraries of 16S rRNA genes cloned from methanogenic oil degrading microcosms amended with North Sea crude oil and inoculated with estuarine sediment indicated that bacteria from the genera Smithella (Deltaproteobacteria, Syntrophaceace) and Marinobacter sp. (Gammaproteobacteria) were enriched during degradation. Growth yields and doubling times (36 days for both Smithella and Marinobacter) were determined using qPCR and quantitative data on alkanes, which were the predominant hydrocarbons degraded. The growth yield of the Smithella sp. [0.020 g(cell-C)/g(alkane-C)], assuming it utilized all alkanes removed was consistent with yields of bacteria that degrade hydrocarbons and other organic compounds in methanogenic consortia. Over 450 days of incubation predominance and exponential growth of Smithella was coincident with alkane removal and exponential accumulation of methane. This growth is consistent with Smithella's occurrence in near surface anoxic hydrocarbon degrading systems and their complete oxidation of crude oil alkanes to acetate and/or hydrogen in syntrophic partnership with methanogens in such systems. The calculated growth yield of the Marinobacter sp., assuming it grew on alkanes, was [0.0005 g(cell-C)/g(alkane-C)] suggesting that it played a minor role in alkane degradation. The dominant methanogens were hydrogenotrophs (Methanocalculus spp. from the Methanomicrobiales). Enrichment of hydrogen-oxidizing methanogens relative to acetoclastic methanogens was consistent with syntrophic acetate oxidation measured in methanogenic crude oil degrading enrichment cultures. qPCR of the Methanomicrobiales indicated growth characteristics consistent with measured rates of methane production and growth in partnership with Smithella. PMID:21914097

  16. [Dendrobium officinale stereoscopic cultivation method].

    PubMed

    Si, Jin-Ping; Dong, Hong-Xiu; Liao, Xin-Yan; Zhu, Yu-Qiu; Li, Hui

    2014-12-01

    The study is aimed to make the most of available space of Dendrobium officinale cultivation facility, reveal the yield and functional components variation of stereoscopic cultivated D. officinale, and improve quality, yield and efficiency. The agronomic traits and yield variation of stereoscopic cultivated D. officinale were studied by operating field experiment. The content of polysaccharide and extractum were determined by using phenol-sulfuric acid method and 2010 edition of "Chinese Pharmacopoeia" Appendix X A. The results showed that the land utilization of stereoscopic cultivated D. officinale increased 2.74 times, the stems, leaves and their total fresh or dry weight in unit area of stereoscopic cultivated D. officinale were all heavier than those of the ground cultivated ones. There was no significant difference in polysaccharide content between stereoscopic cultivation and ground cultivation. But the extractum content and total content of polysaccharide and extractum were significantly higher than those of the ground cultivated ones. In additional, the polysaccharide content and total content of polysaccharide and extractum from the top two levels of stereoscopic culture matrix were significantly higher than that of the ones from the other levels and ground cultivation. Steroscopic cultivation can effectively improves the utilization of space and yield, while the total content of polysaccharides and extractum were significantly higher than that of the ground cultivated ones. The significant difference in Dendrobium polysaccharides among the plants from different height of stereo- scopic culture matrix may be associated with light factor. PMID:25911804

  17. Methanogenic degradation of lignin-derived monoaromatic compounds by microbial enrichments from rice paddy field soil.

    PubMed

    Kato, Souichiro; Chino, Kanako; Kamimura, Naofumi; Masai, Eiji; Yumoto, Isao; Kamagata, Yoichi

    2015-01-01

    Anaerobic degradation of lignin-derived aromatics is an important metabolism for carbon and nutrient cycles in soil environments. Although there are some studies on degradation of lignin-derived aromatics by nitrate- and sulfate-reducing bacteria, knowledge on their degradation under methanogenic conditions are quite limited. In this study, methanogenic microbial communities were enriched from rice paddy field soil with lignin-derived methoxylated monoaromatics (vanillate and syringate) and their degradation intermediates (protocatechuate, catechol, and gallate) as the sole carbon and energy sources. Archaeal community analysis disclosed that both aceticlastic (Methanosarcina sp.) and hydrogenotrophic (Methanoculleus sp. and Methanocella sp.) methanogens dominated in all of the enrichments. Bacterial community analysis revealed the dominance of acetogenic bacteria (Sporomusa spp.) only in the enrichments on the methoxylated aromatics, suggesting that Sporomusa spp. initially convert vanillate and syringate into protocatechuate and gallate, respectively, with acetogenesis via O-demethylation. As the putative ring-cleavage microbes, bacteria within the phylum Firmicutes were dominantly detected from all of the enrichments, while the dominant phylotypes were not identical between enrichments on vanillate/protocatechuate/catechol (family Peptococcaceae bacteria) and on syringate/gallate (family Ruminococcaceae bacteria). This study demonstrates the importance of cooperation among acetogens, ring-cleaving fermenters/syntrophs and aceticlastic/hydrogenotrophic methanogens for degradation of lignin-derived aromatics under methanogenic conditions. PMID:26399549

  18. Windrow composting mitigated CH4 emissions: characterization of methanogenic and methanotrophic communities in manure management.

    PubMed

    Chen, Ruirui; Wang, Yiming; Wei, Shiping; Wang, Wei; Lin, Xiangui

    2014-12-01

    With increasing livestock breeding, methane (CH4 ) emissions from manure management will increasingly contribute more to atmospheric CH4 concentration. The dynamics of methanogens and methanotrophs have not yet been studied in the manure environment. The current study combines surface CH4 emissions with methanogenic and methanotrophic community analyses from two management practices, windrow composting (WCOM) and solid storage (SSTO). Our results showed that there was an c. 50% reduction of CH4 emissions with WCOM compared with SSTO over a 50-day period. A sharp decrease in the quantities of both methanogens and methanotrophs in WCOM suggested that CH4 mitigation was mainly due to decreased CH4 production rather than increased CH4 oxidation. Pyrosequencing analysis demonstrated that aeration caused a clear shift of dominant methanogens in the manure, with specifically a significant decrease in Methanosarcina and increase in Methanobrevibacter. The composition of methanogenic community was influenced by manure management and regulated CH4 production. A sharp increase in the quantity of methanotrophs in SSTO suggested that microbial CH4 oxidation is an important sink for the CH4 produced. The increased abundance of Methylococcaceae in SSTO suggested that Type I methanotrophs have an advantage in CH4 oxidation in occupying niches under low CH4 and high O2 conditions. PMID:25135448

  19. Methanogenic activity and diversity in the centre of the Amsterdam Mud Volcano, Eastern Mediterranean Sea.

    PubMed

    Lazar, Cassandre Sara; John Parkes, R; Cragg, Barry A; L'Haridon, Stephane; Toffin, Laurent

    2012-07-01

    Marine mud volcanoes are geological structures emitting large amounts of methane from their active centres. The Amsterdam mud volcano (AMV), located in the Anaximander Mountains south of Turkey, is characterized by intense active methane seepage produced in part by methanogens. To date, information about the diversity or the metabolic pathways used by the methanogens in active centres of marine mud volcanoes is limited. (14)C-radiotracer measurements showed that methylamines/methanol, H(2)/CO(2) and acetate were used for methanogenesis in the AMV. Methylotrophic methanogenesis was measured all along the sediment core, Methanosarcinales affiliated sequences were detected using archaeal 16S PCR-DGGE and mcrA gene libraries, and enrichments of methanogens showed the presence of Methanococcoides in the shallow sediment layers. Overall acetoclastic methanogenesis was higher than hydrogenotrophic methanogenesis, which is unusual for cold seep sediments. Interestingly, acetate porewater concentrations were extremely high in the AMV sediments. This might be the result of organic matter cracking in deeper hotter sediment layers. Methane was also produced from hexadecanes. For the most part, the methanogenic community diversity was in accordance with the depth distribution of the H(2)/CO(2) and acetate methanogenesis. These results demonstrate the importance of methanogenic communities in the centres of marine mud volcanoes. PMID:22458514

  20. Methanogenic degradation of lignin-derived monoaromatic compounds by microbial enrichments from rice paddy field soil

    PubMed Central

    Kato, Souichiro; Chino, Kanako; Kamimura, Naofumi; Masai, Eiji; Yumoto, Isao; Kamagata, Yoichi

    2015-01-01

    Anaerobic degradation of lignin-derived aromatics is an important metabolism for carbon and nutrient cycles in soil environments. Although there are some studies on degradation of lignin-derived aromatics by nitrate- and sulfate-reducing bacteria, knowledge on their degradation under methanogenic conditions are quite limited. In this study, methanogenic microbial communities were enriched from rice paddy field soil with lignin-derived methoxylated monoaromatics (vanillate and syringate) and their degradation intermediates (protocatechuate, catechol, and gallate) as the sole carbon and energy sources. Archaeal community analysis disclosed that both aceticlastic (Methanosarcina sp.) and hydrogenotrophic (Methanoculleus sp. and Methanocella sp.) methanogens dominated in all of the enrichments. Bacterial community analysis revealed the dominance of acetogenic bacteria (Sporomusa spp.) only in the enrichments on the methoxylated aromatics, suggesting that Sporomusa spp. initially convert vanillate and syringate into protocatechuate and gallate, respectively, with acetogenesis via O-demethylation. As the putative ring-cleavage microbes, bacteria within the phylum Firmicutes were dominantly detected from all of the enrichments, while the dominant phylotypes were not identical between enrichments on vanillate/protocatechuate/catechol (family Peptococcaceae bacteria) and on syringate/gallate (family Ruminococcaceae bacteria). This study demonstrates the importance of cooperation among acetogens, ring-cleaving fermenters/syntrophs and aceticlastic/hydrogenotrophic methanogens for degradation of lignin-derived aromatics under methanogenic conditions. PMID:26399549

  1. Molecular diversity and tools for deciphering the methanogen community structure and diversity in freshwater sediments.

    PubMed

    Chaudhary, Prem Prashant; Brablcová, Lenka; Buriánková, Iva; Rulík, Martin

    2013-09-01

    Methanogenic archaeal communities existing in freshwater sediments are responsible for approximately 50 % of the total global emission of methane. This process contributes significantly to global warming and, hence, necessitates interventional control measures to limit its emission. Unfortunately, the diversity and functional interactions of methanogenic populations occurring in these habitats are yet to be fully characterized. Considering several disadvantages of conventional culture-based methodologies, in recent years, impetus is given to molecular biology approaches to determine the community structure of freshwater sedimentary methanogenic archaea. 16S rRNA and methyl coenzyme M reductase (mcrA) gene-based cloning techniques are the first choice for this purpose. In addition, electrophoresis-based (denaturing gradient gel electrophoresis, temperature gradient gel electrophoresis, and terminal restriction fragment length polymorphism) and quantitative real-time polymerase chain reaction techniques have also found extensive applications. These techniques are highly sensitive, rapid, and reliable as compared to traditional culture-dependent approaches. Molecular diversity studies revealed the dominance of the orders Methanomicrobiales and Methanosarcinales of methanogens in freshwater sediments. The present review discusses in detail the status of the diversity of methanogens and the molecular approaches applied in this area of research. PMID:23877581

  2. Methanogenic degradation of petroleum hydrocarbons in subsurface environments remediation, heavy oil formation, and energy recovery.

    PubMed

    Gray, N D; Sherry, A; Hubert, C; Dolfing, J; Head, I M

    2010-01-01

    Hydrocarbons are common constituents of surface, shallow, and deep-subsurface environments. Under anaerobic conditions, hydrocarbons can be degraded to methane by methanogenic microbial consortia. This degradation process is widespread in the geosphere. In comparison with other anaerobic processes, methanogenic hydrocarbon degradation is more sustainable over geological time scales because replenishment of an exogenous electron acceptor is not required. As a consequence, this process has been responsible for the formation of the world's vast deposits of heavy oil, which far exceed conventional oil assets such as those found in the Middle East. Methanogenic degradation is also a potentially important component of attenuation in hydrocarbon contamination plumes. Studies of the organisms, syntrophic partnerships, mechanisms, and geochemical signatures associated with methanogenic hydrocarbon degradation have identified common themes and diagnostic markers for this process in the subsurface. These studies have also identified the potential to engineer methanogenic processes to enhance the recovery of energy assets as biogenic methane from residual oils stranded in petroleum systems. PMID:20602990

  3. Effect of adding carbon fiber textiles to methanogenic bioreactors used to treat an artificial garbage slurry.

    PubMed

    Sasaki, Kengo; Morita, Masahiko; Hirano, Shin-Ichi; Ohmura, Naoya; Igarashi, Yasuo

    2009-08-01

    To compare the performances and microbial populations of methanogenic reactors with and without carbon fiber textiles (CFT), we operated small-scale (200 ml) reactors using a slurry of artificial garbage. For both types of reactors, the organic loading rate (OLR) was stepwisely and rapidly increased in the same manner. Start-up period was shortened by adding CFT. Reactors with CFT showed greater efficiency for removal of suspended solid and volatile suspended solid than reactors without CFT at a long hydraulic retention time (HRT) between 8 and 13 days. The reactors with CFT maintained stable methane production at an OLR of 15.3 g dichromate chemical oxygen demand (CODcr)/l/day and DNAs from microorganisms were highly concentrated in adhering fractions on CFT. As shown by quantitative PCR analysis, the proportions of methanogenic archaea were conserved more than 25% in adhering fractions on CFT in reactors with CFT. By contrast, reactors without CFT showed accumulation of volatile fatty acid and deteriorated at an OLR of 2.4 gCODcr/l/day. Methanogenic proportions dropped to 17.1% in suspended fractions of reactors without CFT. Denaturing gradient gel electrophoresis (DGGE) analysis revealed that all archaeal DGGE bands in both types of reactors were related to methanogens, but more bands were observed in reactors with CFT. Thus the higher performance of reactors with CFT likely reflects the greater abundance of microorganisms and methanogenic diversity. PMID:19619860

  4. Molecular Analysis of Methanogen Richness in Landfill and Marshland Targeting 16S rDNA Sequences

    PubMed Central

    Yadav, Shailendra; Kundu, Sharbadeb; Ghosh, Sankar K.; Maitra, S. S.

    2015-01-01

    Methanogens, a key contributor in global carbon cycling, methane emission, and alternative energy production, generate methane gas via anaerobic digestion of organic matter. The methane emission potential depends upon methanogenic diversity and activity. Since they are anaerobes and difficult to isolate and culture, their diversity present in the landfill sites of Delhi and marshlands of Southern Assam, India, was analyzed using molecular techniques like 16S rDNA sequencing, DGGE, and qPCR. The sequencing results indicated the presence of methanogens belonging to the seventh order and also the order Methanomicrobiales in the Ghazipur and Bhalsawa landfill sites of Delhi. Sequences, related to the phyla Crenarchaeota (thermophilic) and Thaumarchaeota (mesophilic), were detected from marshland sites of Southern Assam, India. Jaccard analysis of DGGE gel using Gel2K showed three main clusters depending on the number and similarity of band patterns. The copy number analysis of hydrogenotrophic methanogens using qPCR indicates higher abundance in landfill sites of Delhi as compared to the marshlands of Southern Assam. The knowledge about “methanogenic archaea composition” and “abundance” in the contrasting ecosystems like “landfill” and “marshland” may reorient our understanding of the Archaea inhabitants. This study could shed light on the relationship between methane-dynamics and the global warming process. PMID:26568700

  5. Structure and Function of Methanogens along a Short-Term Restoration Chronosequence in the Florida Everglades▿

    PubMed Central

    Smith, Jason M.; Castro, Hector; Ogram, Andrew

    2007-01-01

    The removal of plants and soil to bedrock to eradicate exotic invasive plants within the Hole-in-the-Donut (HID) region, part of the Everglades National Park (Florida), presented a unique opportunity to study the redevelopment of soil and the associated microbial communities in the context of short-term primary succession and ecosystem restoration. The goal of this study was to identify relationships between soil redevelopment and activity and composition of methanogenic assemblages in HID soils. Methane production potentials indicated a general decline in methanogenic activity with restoration age. Microcosm incubations strongly suggested hydrogenotrophic methanogenesis as the most favorable pathway for methane formation in HID soils from all sites. Culture-independent techniques targeting methyl coenzyme M reductase genes (mcrA) were used to assess the dynamics of methanogenic assemblages. Clone libraries were dominated by sequences related to hydrogenotrophic methanogens of the orders Methanobacteriales and Methanococcales and suggested a general decline in the relative abundance of Methanobacteriales mcrA with time since restoration. Terminal restriction fragment length polymorphism analysis indicated methanogenic assemblages remain relatively stable between wet and dry seasons. Interestingly, analysis of soils across the restoration chronosequence indicated a shift in Methanobacteriales populations with restoration age, suggesting genotypic shifts due to site-specific factors. PMID:17449688

  6. Cultivating an entrepreneurial mindset.

    PubMed

    Matheson, Sandra A

    2013-01-01

    Now as never before, familiar challenges require bold, novel approaches. Registered dietitians will benefit by cultivating an entrepreneurial mindset that involves being comfortable with uncertainty, learning to take calculated risks, and daring to just try it. An entrepreneur is someone who takes risks to create something new, usually in business. But the entrepreneurial mindset is available to anyone prepared to rely only on their own abilities for their economic security and expect no opportunity without first creating value for others. PMID:24018008

  7. Reductive dehalogenation of chlorinated benzenes and toluenes under methanogenic conditions.

    PubMed Central

    Ramanand, K; Balba, M T; Duffy, J

    1993-01-01

    The anaerobic metabolism of chlorinated benzenes and toluenes was evaluated in soil slurry microcosms under methanogenic conditions. A mixture of hexachlorobenzene, pentachlorobenzene, and 1,2,4-trichlorobenzene (TCB) in soil slurries was biotransformed through sequential reductive dechlorination to chlorobenzene (CB). The metabolic pathway for hexachlorobenzene and pentachlorobenzene decay proceeded via 1,2,3,4-tetrachlorobenzene (TTCB)-->1,2,3-TCB + 1,2,4-TCB-->1,2-dichlorobenzene (DCB) + 1,4-DCB-->CB. In a mineral salts medium, the CB-adapted soil microorganisms dehalogenated individual 1,2,4,5-TTCB, 1,2,3,4-TTCB, 1,2,3-TCB, and 1,2,4-TCB but not 1,2,3,5-TTCB or 1,3,5-TCB. Similarly, a mixture of 2,3,6-trichlorotoluene (TCT), 2,5-dichlorotoluene (DCT), and 3,4-DCT was reductively dechlorinated in soil slurries to predominantly toluene and small amounts of 2-, 3-, and 4-chlorotoluene (CT). Toluene was further degraded. When tested individually in a mineral salts medium, the CT-adapted soil microorganisms dechlorinated several TCT and DCT isomers. Key metabolic routes for TCTs followed: 2,3,6-TCT-->2,5-DCT-->2-CT-->toluene; 2,4,5-TCT-->2,5-DCT + 3,4-DCT-->3-CT + 4-CT-->toluene. Among DCTs tested, 2,4-DCT and 3,4-DCT were dechlorinated via the removal of o- and m-chlorine, respectively, to 4-CT and subsequently to toluene via p-chlorine removal. Likewise, 2,5-DCT was dechlorinated via 2-CT to toluene. Evidently, microorganisms capable of removing o-, m-, and p-chlorines are present in the soil system, as reflected by the dechlorination of different isomers of CBs and CTs to CB and toluene, respectively. These findings help clarify the metabolic fate of chlorinated benzenes and toluenes in anaerobic environments. PMID:8250553

  8. Transduction-like gene transfer in the methanogen Methanococcus voltae

    NASA Technical Reports Server (NTRS)

    Bertani, G.

    1999-01-01

    Strain PS of Methanococcus voltae (a methanogenic, anaerobic archaebacterium) was shown to generate spontaneously 4.4-kbp chromosomal DNA fragments that are fully protected from DNase and that, upon contact with a cell, transform it genetically. This activity, here called VTA (voltae transfer agent), affects all markers tested: three different auxotrophies (histidine, purine, and cobalamin) and resistance to BES (2-bromoethanesulfonate, an inhibitor of methanogenesis). VTA was most effectively prepared by culture filtration. This process disrupted a fraction of the M. voltae cells (which have only an S-layer covering their cytoplasmic membrane). VTA was rapidly inactivated upon storage. VTA particles were present in cultures at concentrations of approximately two per cell. Gene transfer activity varied from a minimum of 2 x 10(-5) (BES resistance) to a maximum of 10(-3) (histidine independence) per donor cell. Very little VTA was found free in culture supernatants. The phenomenon is functionally similar to generalized transduction, but there is no evidence, for the time being, of intrinsically viral (i.e., containing a complete viral genome) particles. Consideration of VTA DNA size makes the existence of such viral particles unlikely. If they exist, they must be relatively few in number;perhaps they differ from VTA particles in size and other properties and thus escaped detection. Digestion of VTA DNA with the AluI restriction enzyme suggests that it is a random sample of the bacterial DNA, except for a 0.9-kbp sequence which is amplified relative to the rest of the bacterial chromosome. A VTA-sized DNA fraction was demonstrated in a few other isolates of M. voltae.

  9. Transformation of toluene and benzene by mixed methanogenic cultures.

    PubMed Central

    Grbić-Galić, D; Vogel, T M

    1987-01-01

    The aromatic hydrocarbons toluene and benzene were anaerobically transformed by mixed methanogenic cultures derived from ferulic acid-degrading sewage sludge enrichments. In most experiments, toluene or benzene was the only semicontinuously supplied carbon and energy source in the defined mineral medium. No exogenous electron acceptors other than CO2 were present. The cultures were fed 1.5 to 30 mM unlabeled or 14C-labeled aromatic substrates (ring-labeled toluene and benzene or methyl-labeled toluene). Gas production from unlabeled substrates and 14C activity distribution in products from the labeled substrates were monitored over a period of 60 days. At least 50% of the substrates were converted to CO2 and methane (greater than 60%). A high percentage of 14CO2 was recovered from the methyl group-labeled toluene, suggesting nearly complete conversion of the methyl group to CO2 and not to methane. However, a low percentage of 14CO2 was produced from ring-labeled toluene or from benzene, indicating incomplete conversion of the ring carbon to CO2. Anaerobic transformation pathways for unlabeled toluene and benzene were studied with the help of gas chromatography-mass spectrometry. The intermediates detected are consistent with both toluene and benzene degradation via initial oxidation by ring hydroxylation or methyl oxidation (toluene), which would result in the production of phenol, cresols, or aromatic alcohol. Additional reactions, such as demethylation and ring reduction, are also possible. Tentative transformation sequences based upon the intermediates detected are discussed. PMID:3105454

  10. Transduction-Like Gene Transfer in the Methanogen Methanococcus voltae

    PubMed Central

    Bertani, Giuseppe

    1999-01-01

    Strain PS of Methanococcus voltae (a methanogenic, anaerobic archaebacterium) was shown to generate spontaneously 4.4-kbp chromosomal DNA fragments that are fully protected from DNase and that, upon contact with a cell, transform it genetically. This activity, here called VTA (voltae transfer agent), affects all markers tested: three different auxotrophies (histidine, purine, and cobalamin) and resistance to BES (2-bromoethanesulfonate, an inhibitor of methanogenesis). VTA was most effectively prepared by culture filtration. This process disrupted a fraction of the M. voltae cells (which have only an S-layer covering their cytoplasmic membrane). VTA was rapidly inactivated upon storage. VTA particles were present in cultures at concentrations of approximately two per cell. Gene transfer activity varied from a minimum of 2 × 10−5 (BES resistance) to a maximum of 10−3 (histidine independence) per donor cell. Very little VTA was found free in culture supernatants. The phenomenon is functionally similar to generalized transduction, but there is no evidence, for the time being, of intrinsically viral (i.e., containing a complete viral genome) particles. Consideration of VTA DNA size makes the existence of such viral particles unlikely. If they exist, they must be relatively few in number;perhaps they differ from VTA particles in size and other properties and thus escaped detection. Digestion of VTA DNA with the AluI restriction enzyme suggests that it is a random sample of the bacterial DNA, except for a 0.9-kbp sequence which is amplified relative to the rest of the bacterial chromosome. A VTA-sized DNA fraction was demonstrated in a few other isolates of M. voltae. PMID:10321998

  11. Analysis of dsDNA and RNA viromes in methanogenic digesters reveals novel viral genetic diversity.

    PubMed

    Calusinska, Magdalena; Marynowska, Martyna; Goux, Xavier; Lentzen, Esther; Delfosse, Philippe

    2016-04-01

    Although viruses are not the key players of the anaerobic digestion process, they may affect the dynamics of bacterial and archaeal populations involved in biogas production. Until now viruses have received very little attention in this specific habitat; therefore, as a first step towards their characterization, we optimized a virus filtration protocol from anaerobic sludge. Afterwards, to assess dsDNA and RNA viral diversity in sludge samples from nine different reactors fed either with waste water, agricultural residues or solid municipal waste plus agro-food residues, we performed metagenomic analyses. As a result we showed that, while the dsDNA viromes (21 assigned families in total) were dominated by dsDNA phages of the order Caudovirales, RNA viruses (14 assigned families in total) were less diverse and were for the main part plant-infecting viruses. Interestingly, less than 2% of annotated contigs were assigned as putative human and animal pathogens. Our study greatly extends the existing view of viral genetic diversity in methanogenic reactors and shows that these viral assemblages are distinct not only among the reactor types but also from nearly 30 other environments already studied, including the human gut, fermented food, deep sea sediments and other aquatic habitats. PMID:26568175

  12. On the Cultivation of Innovative Talents in Colleges and Universities

    ERIC Educational Resources Information Center

    Yu, Changli; Jia, Hongchun

    2009-01-01

    It is the sure pursuit for the Colleges and Universities to cultivate the innovative talents for the society. The cultivation of innovative talents in Colleges and Universities plays a crucial role not only in economic and social development, but also in schools' and personal development. The internal quality of innovative talents includes the…

  13. Cultivating Discontinuity: Pentecostal Pedagogies of Yielding and Control

    ERIC Educational Resources Information Center

    Brahinsky, Josh

    2013-01-01

    Exploring missionary study at an Assemblies of God Bible college through ethnography and training manuals demonstrates systematic pedagogies that cultivate sensory capabilities encouraging yielding, opening to rupture, and constraint. Ritual theory and the Anthropology of Christianity shift analytic scales to include "cultivation," a…

  14. The molecular genetics of cultivated mushrooms.

    PubMed

    Whiteford, J R; Thurston, C F

    2000-01-01

    The types, economic significance and methods of production of the principal cultivated mushrooms are described in outline. These organisms are all less than ideal for conventional genetic analysis and breeding, so molecular methods afford a particular opportunity to advance our understanding of their biology and potentially give the prospect of improvement by gene manipulation. The sequences described are limited to those found in GenBank by August 1999. The gene sequences isolated from the white button mushroom Agaricus bisporus, the shiitake Lentinula edodes, the oyster mushrooms Pleurotus spp., the paddy straw mushroom Volvariella volvacea and the enotake Flammulina velutipes are described. The largest group are genes from A. bisporus, which includes 29 for intracellular proteins and 12 for secreted proteins. In comparison, only a total of 26 sequences can be reported for the other cultivated species. A. bisporus is also the only cultivated species for which molecular karyotyping is already supported by reliable markers for all 13 of its chromosomes. PMID:10907549

  15. Genome Sequence of “Candidatus Methanomassiliicoccus intestinalis” Issoire-Mx1, a Third Thermoplasmatales-Related Methanogenic Archaeon from Human Feces

    PubMed Central

    Borrel, Guillaume; Harris, Hugh M. B.; Parisot, Nicolas; Gaci, Nadia; Tottey, William; Mihajlovski, Agnès; Deane, Jennifer; Gribaldo, Simonetta; Bardot, Olivier; Peyretaillade, Eric; Peyret, Pierre; O’Toole, Paul W.

    2013-01-01

    “Candidatus Methanomassiliicoccus intestinalis” Issoire-Mx1 is a methanogenic archaeon found in the human gut and is a representative of the novel order of methanogens related to Thermoplasmatales. Its complete genome sequence is presented here. PMID:23846268

  16. Fermentation enhancement of methanogenic archaea consortia from an Illinois basin coalbed via DOL emulsion nutrition.

    PubMed

    Xiao, Dong; Peng, Su-Ping; Wang, En-Yuan

    2015-01-01

    Microbially enhanced coalbed methane technology must be used to increase the methane content in mining and generate secondary biogenic gas. In this technology, the metabolic processes of methanogenic consortia are the basis for the production of biomethane from some of the organic compounds in coal. Thus, culture nutrition plays an important role in remediating the nutritional deficiency of a coal seam. To enhance the methane production rates for microorganism consortia, different types of nutrition solutions were examined in this study. Emulsion nutrition solutions containing a novel nutritional supplement, called dystrophy optional modification latex, increased the methane yield for methanogenic consortia. This new nutritional supplement can help methanogenic consortia form an enhanced anaerobic environment, optimize the microbial balance in the consortia, and improve the methane biosynthesis rate. PMID:25884952

  17. Fermentation Enhancement of Methanogenic Archaea Consortia from an Illinois Basin Coalbed via DOL Emulsion Nutrition

    PubMed Central

    Xiao, Dong; Peng, Su-Ping; Wang, En-Yuan

    2015-01-01

    Microbially enhanced coalbed methane technology must be used to increase the methane content in mining and generate secondary biogenic gas. In this technology, the metabolic processes of methanogenic consortia are the basis for the production of biomethane from some of the organic compounds in coal. Thus, culture nutrition plays an important role in remediating the nutritional deficiency of a coal seam. To enhance the methane production rates for microorganism consortia, different types of nutrition solutions were examined in this study. Emulsion nutrition solutions containing a novel nutritional supplement, called dystrophy optional modification latex, increased the methane yield for methanogenic consortia. This new nutritional supplement can help methanogenic consortia form an enhanced anaerobic environment, optimize the microbial balance in the consortia, and improve the methane biosynthesis rate. PMID:25884952

  18. Toward the identification of methanogenic archaeal groups as targets of methane mitigation in livestock animalsr

    PubMed Central

    St-Pierre, Benoit; Cersosimo, Laura M.; Ishaq, Suzanne L.; Wright, André-Denis G.

    2015-01-01

    In herbivores, enteric methane is a by-product from the digestion of plant biomass by mutualistic gastrointestinal tract (GIT) microbial communities. Methane is a potent greenhouse gas that is not assimilated by the host and is released into the environment where it contributes to climate change. Since enteric methane is exclusively produced by methanogenic archaea, the investigation of mutualistic methanogen communities in the GIT of herbivores has been the subject of ongoing research by a number of research groups. In an effort to uncover trends that would facilitate the development of efficient methane mitigation strategies for livestock species, we have in this review summarized and compared currently available results from published studies on this subject. We also offer our perspectives on the importance of pursuing current research efforts on the sequencing of gut methanogen genomes, as well as investigating their cellular physiology and interactions with other GIT microorganisms. PMID:26284054

  19. Factors influencing the degradation of garbage in methanogenic bioreactors and impacts on biogas formation.

    PubMed

    Morita, Masahiko; Sasaki, Kengo

    2012-05-01

    Anaerobic digestion of garbage is attracting much attention because of its application in waste volume reduction and the recovery of biogas for use as an energy source. In this review, various factors influencing the degradation of garbage and the production of biogas are discussed. The surface hydrophobicity and porosity of supporting materials are important factors in retaining microorganisms such as aceticlastic methanogens and in attaining a higher degradation of garbage and a higher production of biogas. Ammonia concentration, changes in environmental parameters such as temperature and pH, and adaptation of microbial community to ammonia have been related to ammonia inhibition. The effects of drawing electrons from the methanogenic community and donating electrons into the methanogenic community on methane production have been shown in microbial fuel cells and bioelectrochemical reactors. The influences of trace elements, phase separation, and co-digestion are also summarized in this review. PMID:22395906

  20. Investigation into the effect of high concentrations of volatile fatty acids in anaerobic digestion on methanogenic communities

    SciTech Connect

    Franke-Whittle, Ingrid H.; Walter, Andreas; Ebner, Christian; Insam, Heribert

    2014-11-15

    Highlights: • Different methanogenic communities in mesophilic and thermophilic reactors. • High VFA levels do not cause major changes in archaeal communities. • Real-time PCR indicated greater diversity than ANAEROCHIP microarray. - Abstract: A study was conducted to determine whether differences in the levels of volatile fatty acids (VFAs) in anaerobic digester plants could result in variations in the indigenous methanogenic communities. Two digesters (one operated under mesophilic conditions, the other under thermophilic conditions) were monitored, and sampled at points where VFA levels were high, as well as when VFA levels were low. Physical and chemical parameters were measured, and the methanogenic diversity was screened using the phylogenetic microarray ANAEROCHIP. In addition, real-time PCR was used to quantify the presence of the different methanogenic genera in the sludge samples. Array results indicated that the archaeal communities in the different reactors were stable, and that changes in the VFA levels of the anaerobic digesters did not greatly alter the dominating methanogenic organisms. In contrast, the two digesters were found to harbour different dominating methanogenic communities, which appeared to remain stable over time. Real-time PCR results were inline with those of microarray analysis indicating only minimal changes in methanogen numbers during periods of high VFAs, however, revealed a greater diversity in methanogens than found with the array.

  1. Quantitative analysis of ruminal methanogenic microbial populations in beef cattle divergent in phenotypic residual feed intake (RFI) offered contrasting diets

    PubMed Central

    2014-01-01

    Background Methane (CH4) emissions in cattle are an undesirable end product of rumen methanogenic fermentative activity as they are associated not only with negative environmental impacts but also with reduced host feed efficiency. The aim of this study was to quantify total and specific rumen microbial methanogenic populations in beef cattle divergently selected for residual feed intake (RFI) while offered (i) a low energy high forage (HF) diet followed by (ii) a high energy low forage (LF) diet. Ruminal fluid was collected from 14 high (H) and 14 low (L) RFI animals across both dietary periods. Quantitative real time PCR (qRT-PCR) analysis was conducted to quantify the abundance of total and specific rumen methanogenic microbes. Spearman correlation analysis was used to investigate the association between the relative abundance of methanogens and animal performance, rumen fermentation variables and diet digestibility. Results Abundance of methanogens, did not differ between RFI phenotypes. However, relative abundance of total and specific methanogen species was affected (P < 0.05) by diet type, with greater abundance observed while animals were offered the LF compared to the HF diet. Conclusions These findings suggest that differences in abundance of specific rumen methanogen species may not contribute to variation in CH4 emissions between efficient and inefficient animals, however dietary manipulation can influence the abundance of total and specific methanogen species. PMID:25276350

  2. Detection and Isolation Techniques for Methanogens from Microbial Mats (in the El Tatio Geyser Field, Chile)

    NASA Astrophysics Data System (ADS)

    Pearson, E. Z.; Franks, M. A.; Bennett, P.

    2010-12-01

    Isolating methanogenic archea from an extreme environment such as El Tatio (high altitude, arid climate) gives insight to the methanogenic taxas able to adapt and grow under extreme conditions. The hydrothermal waters at El Tatio geyser field demonstrate extreme geochemical conditions, with discharge water from springs and geysers at local boiling temperature (85° C) with high levels of arsenic and low DIC levels. Despite these challenges, many of El Tatio’s hundred plus hydrothermal features host extensive microbial mat communities, many showing evidence of methanogenesis. When trying to isolate methanogens unique to this area, various approaches and techniques were used. To detect the presence of methanogens in samples taken from the field, dissolved methane concentrations were determined via gas chromatography (GC) analysis. Samples were then selected for culturing and most probable number (MPN) enumeration, where growth was assessed using both methane production and observations of fluorescence under UV light. PCR was used to see if the archeal DNA was apparent directly from the field, and shotgun cloning was done to determine phylogenetic affiliation. Several culturing techniques were carried out in an attempt to isolate methanogens from samples that showed evidence of methanogenesis. The slant culturing method was used because of the increased surface area for colonization combined with the relative ease of keeping anaerobic. After a few weeks, when colonies were apparent, some were aseptically selected and inoculated to observe growth in a liquid media containing ampicillin to inhibit bacterial growth. Culturing techniques proved successful after inoculation, showing a slow growth of methanogens via GC and autofluorescence. Further PCR tests and subsequent sequencing were done to confirm and identify isolates.

  3. Distribution, Activities, and Interactions of Methanogens and Sulfate-Reducing Prokaryotes in the Florida Everglades

    PubMed Central

    Bae, Hee-Sung; Holmes, M. Elizabeth; Chanton, Jeffrey P.; Reddy, K. Ramesh

    2015-01-01

    To gain insight into the mechanisms controlling methanogenic pathways in the Florida Everglades, the distribution and functional activities of methanogens and sulfate-reducing prokaryotes (SRPs) were investigated in soils (0 to 2 or 0 to 4 cm depth) across the well-documented nutrient gradient in the water conservation areas (WCAs) caused by runoff from the adjacent Everglades Agricultural Area. The methyl coenzyme M reductase gene (mcrA) sequences that were retrieved from WCA-2A, an area with relatively high concentrations of SO42− (≥39 μM), indicated that methanogens inhabiting this area were broadly distributed within the orders Methanomicrobiales, Methanosarcinales, Methanocellales, Methanobacteriales, and Methanomassiliicoccales. In more than 3 years of monitoring, quantitative PCR (qPCR) using newly designed group-specific primers revealed that the hydrogenotrophic Methanomicrobiales were more numerous than the Methanosaetaceae obligatory acetotrophs in SO42−-rich areas of WCA-2A, while the Methanosaetaceae were dominant over the Methanomicrobiales in WCA-3A (with relatively low SO42− concentrations; ≤4 μM). qPCR of dsrB sequences also indicated that SRPs are present at greater numbers than methanogens in the WCAs. In an incubation study with WCA-2A soils, addition of MoO42− (a specific inhibitor of SRP activity) resulted in increased methane production rates, lower apparent fractionation factors [αapp; defined as (amount of δ13CO2 + 1,000)/(amount of δ13CH4 + 1,000)], and higher Methanosaetaceae mcrA transcript levels compared to those for the controls without MoO42−. These results indicate that SRPs play crucial roles in controlling methanogenic pathways and in shaping the structures of methanogen assemblages as a function of position along the nutrient gradient. PMID:26276115

  4. Distribution, activities, and interactions of methanogens and sulfate-reducing prokaryotes in the Florida Everglades.

    PubMed

    Bae, Hee-Sung; Holmes, M Elizabeth; Chanton, Jeffrey P; Reddy, K Ramesh; Ogram, Andrew

    2015-11-01

    To gain insight into the mechanisms controlling methanogenic pathways in the Florida Everglades, the distribution and functional activities of methanogens and sulfate-reducing prokaryotes (SRPs) were investigated in soils (0 to 2 or 0 to 4 cm depth) across the well-documented nutrient gradient in the water conservation areas (WCAs) caused by runoff from the adjacent Everglades Agricultural Area. The methyl coenzyme M reductase gene (mcrA) sequences that were retrieved from WCA-2A, an area with relatively high concentrations of SO4 (2-) (≥39 μM), indicated that methanogens inhabiting this area were broadly distributed within the orders Methanomicrobiales, Methanosarcinales, Methanocellales, Methanobacteriales, and Methanomassiliicoccales. In more than 3 years of monitoring, quantitative PCR (qPCR) using newly designed group-specific primers revealed that the hydrogenotrophic Methanomicrobiales were more numerous than the Methanosaetaceae obligatory acetotrophs in SO4 (2-)-rich areas of WCA-2A, while the Methanosaetaceae were dominant over the Methanomicrobiales in WCA-3A (with relatively low SO4 (2-) concentrations; ≤4 μM). qPCR of dsrB sequences also indicated that SRPs are present at greater numbers than methanogens in the WCAs. In an incubation study with WCA-2A soils, addition of MoO4 (2-) (a specific inhibitor of SRP activity) resulted in increased methane production rates, lower apparent fractionation factors [αapp; defined as (amount of δ(13)CO2 + 1,000)/(amount of δ(13)CH4 + 1,000)], and higher Methanosaetaceae mcrA transcript levels compared to those for the controls without MoO4 (2-). These results indicate that SRPs play crucial roles in controlling methanogenic pathways and in shaping the structures of methanogen assemblages as a function of position along the nutrient gradient. PMID:26276115

  5. Complete Genome Sequence of the Genetically Tractable Hydrogenotrophic Methanogen Methanococcus maripaludis

    SciTech Connect

    Hendrickson, E. L.; Kaul, R.; Zhou, Y.; Bovee, D.; Chapman, P.; Chung, J.; DeMarcario, E. Conway; Dodsworth, J. A.; Gillett, W.; Graham, D. E.; Hackett, M.; Haydock, A. K.; Kang, A.; Land, Miriam L; Levy, R.; Lie, T. J.; Major, T. A.; Moore, B. C.; Porat, I.; Palmeriri, A.; Rouse, G.; Saenphimmachak, C.; Soll, D; Van Dien, S.; Wang, T.; Whitman, W. B.; Xia, Q.; Zhang, Y.; Larimer, Frank W; Olson, M. V.; Leigh, J. A.

    2004-01-01

    The genome sequence of the genetically tractable, mesophilic, hydrogenotrophic methanogen Methanococcus maripaludis contains 1,722 protein-coding genes in a single circular chromosome of 1,661,137 bp. Of the protein-coding genes (open reading frames [ORFs]), 44% were assigned a function, 48% were conserved but had unknown or uncertain functions, and 7.5% (129 ORFs) were unique to M. maripaludis. Of the unique ORFs, 27 were confirmed to encode proteins by the mass spectrometric identification of unique peptides. Genes for most known functions and pathways were identified. For example, a full complement of hydrogenases and methanogenesis enzymes was identified, including eight selenocysteine-containing proteins, with each being paralogous to a cysteine-containing counterpart. At least 59 proteins were predicted to contain iron-sulfur centers, including ferredoxins, polyferredoxins, and subunits of enzymes with various redox functions. Unusual features included the absence of a Cdc6 homolog, implying a variation in replication initiation, and the presence of a bacterial-like RNase HI as well as an RNase HII typical of the Archaea. The presence of alanine dehydrogenase and alanine racemase, which are uniquely present among the Archaea, explained the ability of the organism to use L- and D-alanine as nitrogen sources. Features that contrasted with the related organism Methanocaldococcus jannaschii included the absence of inteins, even though close homologs of most intein-containing proteins were encoded. Although two-thirds of the ORFs had their highest Blastp hits in Methanocaldococcus jannaschii, lateral gene transfer or gene loss has apparently resulted in genes, which are often clustered, with top Blastp hits in more distantly related groups.

  6. Hydrobiogeochemical controls on a low-carbon emitting energy extraction mechanism: exploring methanogenic crude oil biodegradation

    NASA Astrophysics Data System (ADS)

    Shelton, Jenna; McIntosh, Jennifer; Akob, Denise; Spear, John; Warwick, Peter; McCray, John

    2016-04-01

    Exploiting naturally-occurring microbial communities in the deep subsurface could help mitigate the effects of CO2 emissions to the atmosphere. These microbial communities, a combination of methanogens and syntrophic bacteria, can perform methanogenic crude oil biodegradation, namely the conversion of crude oil to natural gas, and have also been detected in biodegraded, methanogenic reservoirs. These microbes could target residual crude oil, a high-carbon, hard-to-obtain fossil fuel source, and convert it to natural gas, effectively "producing" a lower CO2 per BTU fuel source. Yet, little is known about what geochemical parameters are driving microbial population dynamics in biodegraded, methanogenic oil reservoirs, and how the presence of specific microbial communities may impact methanogenic crude oil biodegradation. To investigate methanogenic crude oil biodegradation, 22 wells along a subsurface hydrogeochemical gradient in the southeastern USA were sampled for DNA analysis of the microbial community, and geochemical analysis of produced water and crude oil. A statistical comparison of microbial community structure to formation fluid geochemical parameters, amount of crude oil biodegradation, and relative extent of methanogenesis revealed that relative degree of biodegradation (high, medium, or low), chloride concentration (550 mM to 2100 mM), well depth (393 m to 1588 m), and spatial location within the reservoir (i.e., oil field location) are the major drivers of microbial diversity. There was no statistical evidence for correlation between extent of methanogenesis and the subsurface community composition. Despite the dominance of methanogens in these sampled wells, methanogenic activity was not predicted solely based on the microbial community composition. Crude oil biodegradation, however, correlates with both community composition and produced water geochemistry, suggesting a co-linear system and implying that microbial communities associated with degree

  7. Growth of Methanogens on a Mars Soil Simulant Under Simulated Martian Conditions

    NASA Astrophysics Data System (ADS)

    Kral, Timothy A.; Bekkum, Curtis R.; McKay, Christopher P.

    2004-06-01

    Due to the hostile conditions at the surface, any life forms existing on Mars today would most likely inhabit a subsurface environment where conditions are potentially wetter and warmer, but organic compounds may be lacking and light energy for photosynthesis would be absent. Methanogens, members of the domain Archaea, are microorganisms from planet Earth that can grow under these relatively extreme conditions. We have demonstrated that certain methanogenic species can indeed grow on a Mars soil simulant, JSC Mars-1, with limited amounts of water, under conditions approaching a possible subsurface environment on Mars.

  8. Potential shuttle vectors based on the methanogen plasmid pME2001

    SciTech Connect

    Meile, L.; Reeve, J.N.

    1985-01-01

    Methane is produced by anaerobic archaebacteria known as methanogens. Currently the only available plasmid from a methanogen is pME2001. The authors incorporated pME2001 into plasmids which should be capable of replication in a range of microbial host species. Plasmid pET2411, a recombinant plasmid formed by joining pBR322 to pME2001, directs the synthesis of pME2001 encoded polypeptides in Escherichia coli but cannot replicate in E. coli in the absence of E. coli DNA polymerase I. 23 references, 3 figures, 1 table.

  9. Changes in methanogenic substrate utilization and communities with depth in a salt-marsh, creek sediment in southern England

    NASA Astrophysics Data System (ADS)

    John Parkes, R.; Brock, Fiona; Banning, Natasha; Hornibrook, Edward R. C.; Roussel, Erwan G.; Weightman, Andrew J.; Fry, John C.

    2012-01-01

    A combined biogeochemical and molecular genetic study of creek sediments (down to 65 cm depth) from Arne Peninsula salt-marsh (Dorset, UK) determined the substrates used for methanogenesis and the distribution of the common methanogens, Methanosarcinales and Methanomicrobiales capable of metabolising these substrates. Methane concentrations increased by 11 cm, despite pore water sulphate not being removed until 45 cm. Neither upward methane diffusion or anaerobic oxidation of methane seemed to be important in this zone. In the near-surface sulphate-reduction zone (5-25 cm) turnover time to methane for the non-competitive methanogenic substrate trimethylamine was most rapid (80 days), and were much longer for acetate (7900 days), methanol (40,500 days) and bicarbonate (361,600 days). Methylamine-utilizing Methanosarcinales were the dominant (60-95%) methanogens in this zone. In deeper sediments rates of methanogenesis from competitive substrates increased substantially, with acetate methanogenic rates becoming ˜100 times greater than H 2/CO 2 methanogenesis below 50 cm. In addition, there was a dramatic change in methanogen diversity with obligate acetate-utilizing, Methanosaeta related sequences being dominant. At a similar depth methanol turnover to methane increased to its most rapid (1700 days). This activity pattern is consistent with deeper methanogen populations (55 cm) being dominated by acetate-utilizing Methanosaeta with H 2/CO 2 and alcohol-utilizing Methanomicrobiales also present. Hence, there is close relationship between the depth distribution of methanogenic substrate utilization and specific methanogens that can utilize these compounds. It is unusual for acetate to be the dominant methanogenic substrate in coastal sediments and δ13C-CH 4 values (-74 to -71‰) were atypical for acetate methanogenesis, suggesting that common stable isotope proxy models may not apply well in this type of dynamic anoxic sediment, with multiple methanogenic substrates.

  10. Complete genome sequence of Methanolinea tarda NOBI-1T, a hydrogenotrophic methanogen isolated from methanogenic digester sludge

    SciTech Connect

    Yamamoto, Kyosuke; Tamaki, Hideyuki; Cadillo-Quiroz, Hinsby; Imachi, Hiroyuki; Kyrpides, Nikos; Woyke, Tanja; Goodwin, Lynne; Zinder, Stephen H.; Kamagata, Yoichi; Liu, Wen -Tso

    2014-09-04

    In this study, we report a 2.0-Mb complete genome sequence of Methanolinea tarda NOBI-1T, a methanogenic archaeon isolated from an anaerobic digested sludge. This is the first genome report of the genus Methanolinea isolate belonging to the family Methanoregulaceae, a recently proposed novel family within the order Methanomicrobiales.

  11. Archaeal Community Changes Associated with Cultivation of Amazon Forest Soil with Oil Palm

    PubMed Central

    Tupinambá, Daiva Domenech; Cantão, Maurício Egídio; Costa, Ohana Yonara Assis; Bergmann, Jessica Carvalho; Kruger, Ricardo Henrique; Kyaw, Cynthia Maria; Barreto, Cristine Chaves; Quirino, Betania Ferraz

    2016-01-01

    This study compared soil archaeal communities of the Amazon forest with that of an adjacent area under oil palm cultivation by 16S ribosomal RNA gene pyrosequencing. Species richness and diversity were greater in native forest soil than in the oil palm-cultivated area, and 130 OTUs (13.7%) were shared between these areas. Among the classified sequences, Thaumarchaeota were predominant in the native forest, whereas Euryarchaeota were predominant in the oil palm-cultivated area. Archaeal species diversity was 1.7 times higher in the native forest soil, according to the Simpson diversity index, and the Chao1 index showed that richness was five times higher in the native forest soil. A phylogenetic tree of unclassified Thaumarchaeota sequences showed that most of the OTUs belong to Miscellaneous Crenarchaeotic Group. Several archaeal genera involved in nutrient cycling (e.g., methanogens and ammonia oxidizers) were identified in both areas, but significant differences were found in the relative abundances of Candidatus Nitrososphaera and unclassified Soil Crenarchaeotic Group (prevalent in the native forest) and Candidatus Nitrosotalea and unclassified Terrestrial Group (prevalent in the oil palm-cultivated area). More studies are needed to culture some of these Archaea in the laboratory so that their metabolism and physiology can be studied. PMID:27006640

  12. The Cultivated Mind: From Mental Mediation to Cultivation.

    ERIC Educational Resources Information Center

    Josephs, Ingrid E.; Fuhrer, Urs

    1998-01-01

    Examines Simmel's principle of cultivation whereby the cultivated mind is constructed through ongoing transactions of people with their cultural environment, cultural forms currently overlooked. Cultural forms result from externalizations of former person-culture transactions. Argues that development is structured through person-culture…

  13. Methanogenic pathway and community structure in a thermophilic anaerobic digestion process of organic solid waste.

    PubMed

    Sasaki, Daisuke; Hori, Tomoyuki; Haruta, Shin; Ueno, Yoshiyuki; Ishii, Masaharu; Igarashi, Yasuo

    2011-01-01

    The methanogenic pathway and microbial community in a thermophilic anaerobic digestion process of organic solid waste were investigated in a continuous-flow stirred-tank reactor using artificial garbage slurry as a feedstock. The decomposition pathway of acetate, a significant precursor of CH(4) and a key intermediate metabolite in the anaerobic digestion process, was analyzed by using stable isotopes. A tracer experiment using (13)C-labeled acetate revealed that approximately 80% of the acetate was decomposed via a non-aceticlastic oxidative pathway, whereas the remainder was converted to methane via an aceticlastic pathway. Archaeal 16S rRNA analyses demonstrated that the hydrogenotrophic methanogens Methanoculleus spp. accounted for >90% of detected methanogens, and the aceticlastic methanogens Methanosarcina spp. were the minor constituents. The clone library targeting bacterial 16S rRNA indicated the predominance of the novel Thermotogales bacterium (relative abundance: ~53%), which is related to anaerobic acetate oxidizer Thermotoga lettingae TMO, although the sequence similarity was low. Uncultured bacteria that phylogenetically belong to municipal solid waste cluster I were also predominant in the microflora (~30%). These results imply that the microbial community in the thermophilic degrading process of organic solid waste consists exclusively of unidentified bacteria, which efficiently remove acetate through a non-aceticlastic oxidative pathway. PMID:20851673

  14. N epsilon-acetyl-beta-lysine: an osmolyte synthesized by methanogenic archaebacteria.

    PubMed Central

    Sowers, K R; Robertson, D E; Noll, D; Gunsalus, R P; Roberts, M F

    1990-01-01

    Methanosarcina thermophila, a nonmarine methanogenic archaebacterium, can grow in a range of saline concentrations. At less than 0.4 M NaCl, Ms. thermophila accumulated glutamate in response to increasing osmotic stress. At greater than 0.4 M NaCl, this organism synthesized a modified beta-amino acid that was identified as N epsilon-acetyl-beta-lysine by NMR spectroscopy and ion-exchange HPLC. This beta-amino acid derivative accumulated to high intracellular concentrations (up to 0.6 M) in Ms. thermophila and in another methanogen examined--Methanogenium cariaci, a marine species. The compound has features that are characteristic of a compatible solute: it is neutrally charged at physiological pH and it is highly soluble. When the cells were grown in the presence of exogenous glycine betaine, a physiological compatible solute, N epsilon-acetyl-beta-lysine synthesis was repressed and glycine betaine was accumulated. N epsilon-acetyl-beta-lysine was synthesized by species from three phylogenetic families when grown in high solute concentrations, suggesting that it may be ubiquitous among the methanogens. The ability to control the biosynthesis of N epsilon-acetyl-beta-lysine in response to extracellular solute concentration indicates that the methanogenic archaebacteria have a unique beta-amino acid biosynthetic pathway that is osmotically regulated. PMID:2123548

  15. Can abundance of methanogen be a good indicator for CH4 flux in soil ecosystems?

    PubMed

    Kim, Jinhyun; Lee, Seung-Hoon; Jang, Inyoung; Jeong, Sangseom; Kang, Hojeong

    2015-12-01

    Methane, which is produced by methanogenic archaea, is the second most abundant carbon compound in the atmosphere. Due to its strong radiative forcing, many studies have been conducted to determine its sources, budget, and dynamics. However, a mechanistic model of methane flux has not been developed thus far. In this study, we attempt to examine the relevance of the abundance of methanogen as a biological indicator of methane flux in three different types of soil ecosystems: permafrost, rice paddy, and mountainous wetland. We measured the annual average methane flux and abundance of methanogen in the soil ecosystems in situ. The correlation between methane flux and the abundance of methanogen exists only under a specific biogeochemical conditions such as SOM of higher than 60%, pH of 5.6-6.4, and water-saturated. Except for these conditions, significant correlations were absent. Therefore, microbial abundance information can be applied to a methane flux model selectively depending on the biogeochemical properties of the soil ecosystem. PMID:26123992

  16. Rumen Methanogenic Genotypes Differ in Abundance According to Host Residual Feed Intake Phenotype and Diet Type

    PubMed Central

    Carberry, CIara A.; Kenny, David A.; Creevey, Christopher J.

    2014-01-01

    Methane is an undesirable end product of rumen fermentative activity because of associated environmental impacts and reduced host feed efficiency. Our study characterized the rumen microbial methanogenic community in beef cattle divergently selected for phenotypic residual feed intake (RFI) while offered a high-forage (HF) diet followed by a low-forage (LF) diet. Rumen fluid was collected from 14 high-RFI (HRFI) and 14 low-RFI (LRFI) animals at the end of both dietary periods. 16S rRNA gene clone libraries were used, and methanogen-specific tag-encoded pyrosequencing was carried out on the samples. We found that Methanobrevibacter spp. are the dominant methanogens in the rumen, with Methanobrevibacter smithii being the most abundant species. Differences in the abundance of Methanobrevibacter smithii and Methanosphaera stadtmanae genotypes were detected in the rumen of animals offered the LF compared to the HF diet while the abundance of Methanobrevibacter smithii genotypes was different between HRFI and LRFI animals irrespective of diet. Our results demonstrate that while a core group of methanogen operational taxonomic units (OTUs) exist across diet and phenotype, significant differences were observed in the distribution of genotypes within those OTUs. These changes in genotype abundance may contribute to the observed differences in methane emissions between efficient and inefficient animals. PMID:24212580

  17. Biokinetic and molecular studies of methanogens in phased anaerobic digestion systems.

    PubMed

    Zamanzadeh, Mirzaman; Parker, Wayne J; Verastegui, Yris; Neufeld, Josh D

    2013-12-01

    The influence of differing operational conditions of two-stage digesters on biokinetic characteristics and communities of methanogenic archaea was evaluated. Operating temperature of each phase influenced the archaeal communities significantly. Also, a strong correlation was observed between community composition and temperature and pH. The maximum specific substrate utilization rates (k max) of acetoclastic methanogens in the mesophilic and thermophilic 1st phases were 11.4 and 22.0 mgCOD mgCOD(-1)d(-1), respectively, whereas significantly lower k max values were estimated for the mesophilic and thermophilic 2nd-phase digesters which were 7.6 and 16.6 mgCOD mgCOD(-1)d(-1), respectively. It appeared that the biokinetic characteristics of the acetoclastic methanogen communities were reliant on digester loading rates. Also, higher temperature dependency coefficients (θ) were observed for the long retention time digesters when compared to the values computed for the 1st-phase digesters. Accordingly, the implementation of two sets of biokinetic parameters for acetoclastic methanogen will improve modeling of phased anaerobic digesters. PMID:24125797

  18. Ruminal fermentation of anti-methanogenic nitrate- and nitro-containing forages in vitro

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Nitrate, 3-nitro-1-propionic acid (NPA), and 3-nitro-1-propanol (NPOH) can accumulate in forages and be poisonous to animals if fed at high enough amounts. These chemicals are also recognized as potent anti-methanogenic compounds, but plants naturally containing these chemicals have been studied li...

  19. Reduction of structural Fe(III) in nontronite by methanogen Methanosarcina barkeri

    NASA Astrophysics Data System (ADS)

    Liu, Deng; Dong, Hailiang; Bishop, Michael E.; Wang, Hongmei; Agrawal, Abinash; Tritschler, Sarah; Eberl, Dennis D.; Xie, Shucheng

    2011-02-01

    Clay minerals and methanogens are ubiquitous and co-exist in anoxic environments, yet it is unclear whether methanogens are able to reduce structural Fe(III) in clay minerals. In this study, the ability of methanogen Methanosarcina barkeri to reduce structural Fe(III) in iron-rich smectite (nontronite NAu-2) and the relationship between iron reduction and methanogenesis were investigated. Bioreduction experiments were conducted in growth medium using three types of substrate: H 2/CO 2, methanol, and acetate. Time course methane production and hydrogen consumption were measured by gas chromatography. M. barkeri was able to reduce structural Fe(III) in NAu-2 with H 2/CO 2 and methanol as substrate, but not with acetate. The extent of bioreduction, as measured by the 1,10-phenanthroline method, was 7-13% with H 2/CO 2 as substrate, depending on nontronite concentration (5-10 g/L). The extent was higher when methanol was used as a substrate, reaching 25-33%. Methanogenesis was inhibited by Fe(III) reduction in the H 2/CO 2 culture, but enhanced when methanol was used. High charge smectite and biogenic silica formed as a result of bioreduction. Our results suggest that methanogens may play an important role in biogeochemical cycling of iron in clay minerals and may have important implications for the global methane budget.

  20. Methanogens couple anaerobic Fe(O) oxidation and CHCl{sub 3} reduction

    SciTech Connect

    Weathres, L.J.; Parkin, G.F.; Novak, P.J.; Alvarez, P.J.J.

    1995-12-01

    Recent studies have shown that chlorinated aliphatic hydrocarbons (CAHs) may be reduced by metallic iron. These studies have focused on abiotic processes, while limited attention has been given to combined microbial and abiotic dechlorination. Methanogenic bacteria can use metallic iron as an energy source, by coupling the anodic dissolution of iron with the consumption of water-derived hydrogen. Hence, under methanogenic conditions in the presence of zero-valent iron, two degradation mechanisms may be important Fe(O) may reduce CAHs abiotically, and Fe(O) may indirectly reduce CAHs via biodehalogenation. Our results using steel wool and methanogenic cell suspension indicate that methanogens coupled the biocorrosion of iron metal and biodehalogenation of CHCl{sub 3} via cometabolism, with water-derived hydrogen acting as energy source. This work demonstrates that, at least under specific conditions, biodehalogenation dominates abiotic mechanisms in aqueous systems containing zero-valent g iron. Further work is required to determine if these results are sustainable in flow-through environments.

  1. Degradation of methanethiol in anaerobic sewers and its correlation with methanogenic activities.

    PubMed

    Sun, Jing; Hu, Shihu; Sharma, Keshab Raj; Ni, Bing-Jie; Yuan, Zhiguo

    2015-02-01

    Methanethiol (MT) is considered one of the predominant odorants in sewer systems. Therefore, understanding MT transformation in sewers is essential to sewer odor assessment and abatement. In this study, we investigated the degradation of MT in laboratory anaerobic sewers. Experiments were carried out in seven anaerobic sewer reactors with biofilms at different stages of development. MT degradation was found to be strongly dependent on the methanogenic activity of sewer biofilms. The MT degradation rate accelerated with the increase of methanogenic activity of sewer biofilms, resulting in MT accumulation (i.e. net production) in sewer reactors with relatively low methanogenic activities, and MT removal in reactors with higher methanogenic activities. A Monod-type kinetic expression was developed to describe MT degradation kinetics in anaerobic sewers, in which the maximum degradation rate was modeled as a function of the maximum methane production rate through a power function. It was also found that MT concentration had a linear relationship with acetate concentration, which may be used for preliminary assessment of MT presence in anaerobic sewers. PMID:25437340

  2. Methanogenic Inhibition by Roxarsone (4-Hydroxy-3-nitrophenylarsonic acid) and Related Aromatic Arsenic Compounds

    PubMed Central

    Sierra-Alvarez, Reyes; Cortinas, Irail; Field, Jim A.

    2009-01-01

    Roxarsone (4-hydroxy-3-nitro-phenylarsonic acid) and p-arsanilic acid (4-aminophenylarsonic acid) are feed additives widely used in the broiler and swine industry. This study evaluated the inhibitory effect of roxarsone, p-arsanilic, and other phenylarsonic compounds on the activity of acetate- and H2-utilizing methanogenic microorganisms. Roxarsone, p-arsanilic, and 4-hydroxy-3-aminophenylarsonic acid (HAPA) inhibited acetoclastic and hydrogenotrophic methanogens when supplemented at concentrations of 1 mM, and their inhibitory effect increased sharply with incubation time. Phenylarsonic acid (1 mM) inhibited acetoclastic but not H2-utilizing methanogens. HAPA, a metabolite from the anaerobic biodegradation of roxarsone, was found to be sensitive to autooxidation by oxygen. The compound (2.6 mM) caused low methanogenic inhibition (only 14.2%) in short-term assays of 12 h when autooxidation was prevented by supplementing HAPA solutions with ascorbate. However, ascorbate-free HAPA solutions underwent spontaneous autooxidation in the presence of oxygen, leading to the formation of highly inhibitory compounds. These results confirm the microbial toxicity of organoarsenic compounds, and they indicate that biotic as well as abiotic transformations can potentially impact the fate and microbial toxicity of these contaminants in the environment. PMID:19889499

  3. Analysis of rumen methanogen diversity in water buffaloes (Bubalus bubalis) under three different diets.

    PubMed

    Franzolin, Raul; St-Pierre, Benoit; Northwood, Korinne; Wright, André-Denis G

    2012-07-01

    The water buffalo (Bubalus bubalis) is a prominent livestock species for the production of milk and meat in many countries. We investigated the diversity of rumen methanogens in Mediterranean water buffaloes maintained in Brazil under different diets: corn silage, grazing pasture, or sugar cane. A total of 467 clones were isolated from three methanogen 16S rRNA gene clone libraries that each represented a distinct feed type. The 467 clones were assigned to 19 species-level operational taxonomic units (OTUs). Four OTUs were represented in all three libraries, eight OTUs were library-specific, six OTUs were found in only the corn silage and pasture grazing libraries, and one OTU was shared only between pasture grazing and sugar cane libraries. We found that Methanobrevibacter-related sequences were the most abundant in the water buffaloes sampled for our analysis, in contrast to previously reported studies showing that Methanomicrobium mobile-like methanogens were the most abundant methanogens in water buffaloes of Murrah and Surti breeds sampled in India. Considering the worldwide distribution of water buffaloes and the likely wide variety of diets provided, our results combined with studies from other groups support that larger scope analyses of microbiomes for this livestock species would provide great insight into the contribution of geographical location, breed, and diet in determining the population structure of rumen microorganisms. PMID:22286379

  4. Complete Genome Sequence of the Methanogen Methanoculleus bourgensis BA1 Isolated from a Biogas Reactor

    PubMed Central

    Maus, Irena; Wibberg, Daniel; Winkler, Anika; Pühler, Alfred; Schnürer, Anna

    2016-01-01

    Methanoculleus bourgensis BA1, a hydrogenotrophic methanogen, was isolated from a laboratory-scale biogas reactor operating under an elevated ammonium concentration. Here, the complete genome sequence of M. bourgensis BA1 is reported. The availability of the BA1 genome sequence enables detailed comparative analyses involving other Methanoculleus spp. representing important members of microbial biogas communities. PMID:27340059

  5. High Concentrations of Methyl Fluoride Affect the Bacterial Community in a Thermophilic Methanogenic Sludge

    PubMed Central

    Hao, Liping; Lü, Fan; Wu, Qing; Shao, Liming; He, Pinjing

    2014-01-01

    To precisely control the application of methyl fluoride (CH3F) for analysis of methanogenic pathways, the influence of 0–10% CH3F on bacterial and archaeal communities in a thermophilic methanogenic sludge was investigated. The results suggested that CH3F acts specifically on acetoclastic methanogenesis. The inhibitory effect stabilized at an initial concentration of 3–5%, with around 90% of the total methanogenic activity being suppressed, and a characteristic of hydrogenotrophic pathway in isotope fractionation was demonstrated under this condition. However, extended exposure (12 days) to high concentrations of CH3F (>3%) altered the bacterial community structure significantly, resulting in increased diversity and decreased evenness, which can be related to acetate oxidation and CH3F degradation. Bacterial clone library analysis showed that syntrophic acetate oxidizing bacteria Thermacetogenium phaeum were highly enriched under the suppression of 10% CH3F. However, the methanogenic community did not change obviously. Thus, excessive usage of CH3F over the long term can change the composition of the bacterial community. Therefore, data from studies involving the use of CH3F as an acetoclast inhibitor should be interpreted with care. Conversely, CH3F has been suggested as a factor to stimulate the enrichment of syntrophic acetate oxidizing bacteria. PMID:24658656

  6. Formation of methane and carbon dioxide from dimethylselenide in anoxic sediments and by a methanogenic bacterium

    USGS Publications Warehouse

    Oremland, Ronald S.; Zehr, Jon P.

    1986-01-01

    Anaerobic San Francisco Bay salt marsh sediments rapidly metabolized [14C]dimethylselenide (DMSe) to 14CH4 and 14CO2. Addition of selective inhibitors (2-bromoethanesulfonic acid or molybdate) to these sediments indicated that both methanogenic and sulfate-respiring bacteria could degrade DMSe to gaseous products. However, sediments taken from the selenium-contaminated Kesterson Wildlife Refuge produced only 14CO2 from [14C]DMSe, implying that methanogens were not important in the Kesterson samples. A pure culture of a dimethylsulfide (DMS)-grown methylotrophic methanogen converted [14C]DMSe to 14CH4 and14CO2. However, the organism could not grow on DMSe. Addition of DMS to either sediments or the pure culture retarded the metabolism of DMSe. This effect appeared to be caused by competitive inhibition, thereby indicating a common enzyme system for DMS and DMSe metabolism. DMSe appears to be degraded as part of the DMS pool present in anoxic environments. These results suggest that methylotrophic methanogens may demethylate methylated forms of other metals and metalloids found in nature.

  7. Peptidolytic Microbial Community of Methanogenic Reactors from two Modified Uasbs of Brewery Industries.

    PubMed

    Díaz, C; Baena, S; Patel, B K C; Fardeau, M L

    2010-07-01

    We studied the peptide-degrading anaerobic communities of methanogenic reactors from two mesophilic full-scale modified upflow anaerobic sludge blanket (UASB) reactors treating brewery wastewater in Colombia. Most probable number (MPN) counts varied between 7.1 x 10(8) and 6.6 × 10(9) bacteria/g volatile suspended solids VSS (Methanogenic Reactor 1) and 7.2 × 10(6) and 6.4 × 10(7) bacteria/g (VSS) (Methanogenic Reactor 2). Metabolites detected in the highest positive MPN dilutions in both reactors were mostly acetate, propionate, isovalerate and, in some cases, negligible concentrations of butyrate. Using the highest positive dilutions of MPN counts, 50 dominant strains were isolated from both reactors, and 12 strains were selected for sequencing their 16S rRNA gene based on their phenotypic characteristics. The small-subunit rRNA gene sequences indicated that these strains were affiliated to the families Propionibacteriaceae, Clostridiaceae and Syntrophomonadaceae in the low G + C gram-positive group and Desulfovibrio spp. in the class δ-Proteobacteria. The main metabolites detected in the highest positive dilutions of MPN and the presence of Syntrophomonadaceae indicate the effect of the syntrophic associations on the bioconversion of these substrates in methanogenic reactors. Additionally, the potential utilization of external electron acceptors for the complete degradation of amino acids by Clostridium strains confirms the relevance of these acceptors in the transformation of peptides and amino acids in these systems. PMID:24031547

  8. Peptidolytic Microbial Community of Methanogenic Reactors from two Modified Uasbs of Brewery Industries

    PubMed Central

    Díaz, C.; Baena, S.; Patel, B.K.C.; Fardeau, M.L.

    2010-01-01

    We studied the peptide-degrading anaerobic communities of methanogenic reactors from two mesophilic full-scale modified upflow anaerobic sludge blanket (UASB) reactors treating brewery wastewater in Colombia. Most probable number (MPN) counts varied between 7.1 x 108 and 6.6 × 109 bacteria/g volatile suspended solids VSS (Methanogenic Reactor 1) and 7.2 × 106 and 6.4 × 107 bacteria/g (VSS) (Methanogenic Reactor 2). Metabolites detected in the highest positive MPN dilutions in both reactors were mostly acetate, propionate, isovalerate and, in some cases, negligible concentrations of butyrate. Using the highest positive dilutions of MPN counts, 50 dominant strains were isolated from both reactors, and 12 strains were selected for sequencing their 16S rRNA gene based on their phenotypic characteristics. The small-subunit rRNA gene sequences indicated that these strains were affiliated to the families Propionibacteriaceae, Clostridiaceae and Syntrophomonadaceae in the low G + C gram-positive group and Desulfovibrio spp. in the class δ-Proteobacteria. The main metabolites detected in the highest positive dilutions of MPN and the presence of Syntrophomonadaceae indicate the effect of the syntrophic associations on the bioconversion of these substrates in methanogenic reactors. Additionally, the potential utilization of external electron acceptors for the complete degradation of amino acids by Clostridium strains confirms the relevance of these acceptors in the transformation of peptides and amino acids in these systems. PMID:24031547

  9. Rumen methanogenic genotypes differ in abundance according to host residual feed intake phenotype and diet type.

    PubMed

    Carberry, Ciara A; Waters, Sinéad M; Waters, Sinead M; Kenny, David A; Creevey, Christopher J

    2014-01-01

    Methane is an undesirable end product of rumen fermentative activity because of associated environmental impacts and reduced host feed efficiency. Our study characterized the rumen microbial methanogenic community in beef cattle divergently selected for phenotypic residual feed intake (RFI) while offered a high-forage (HF) diet followed by a low-forage (LF) diet. Rumen fluid was collected from 14 high-RFI (HRFI) and 14 low-RFI (LRFI) animals at the end of both dietary periods. 16S rRNA gene clone libraries were used, and methanogen-specific tag-encoded pyrosequencing was carried out on the samples. We found that Methanobrevibacter spp. are the dominant methanogens in the rumen, with Methanobrevibacter smithii being the most abundant species. Differences in the abundance of Methanobrevibacter smithii and Methanosphaera stadtmanae genotypes were detected in the rumen of animals offered the LF compared to the HF diet while the abundance of Methanobrevibacter smithii genotypes was different between HRFI and LRFI animals irrespective of diet. Our results demonstrate that while a core group of methanogen operational taxonomic units (OTUs) exist across diet and phenotype, significant differences were observed in the distribution of genotypes within those OTUs. These changes in genotype abundance may contribute to the observed differences in methane emissions between efficient and inefficient animals. PMID:24212580

  10. Methanogenic archaea diversity in hyporheic sediments of a small lowland stream

    NASA Astrophysics Data System (ADS)

    Brablcova, Lenka; Buriánková, Iva; Rulík, Martin

    2015-04-01

    Abundance and diversity of methanogenic archaea were studied at five localities along a longitudinal profile of a Sitka stream (Czech Republic). Samples of hyporheic sediments were collected from two sediment depths (0-25 cm and 25-50 cm) by freeze-core method. Methanogen community was analyzed by fluorescence in situ hybridization (FISH), denaturing gradient gel electrophoresis (DGGE) and sequencing method. The proportion of methanogens to the DAPI-stained cells varied among all localities and depths with an average value 2.08 × 105 per g of dry sediment. A total of 73 bands were detected at 19 different positions on the DGGE gel and the highest methanogen diversity was found at the downstream located sites. Cluster analysis of DGGE image showed three main clusters consisting of localities that differed in the number and similarity of the DGGE bands. Sequencing analysis of representative DGGE bands revealed phylotypes affiliated with members belonging to the orders Methanosarcinales, Methanomicrobiales and Methanocellales. The authors are thankful to the European Social Fund and state budget of the Czech Republic for providing the financial support during this study. This work is a part of the POSTUP II project CZ.1.07/2.3.00/30.0041, which is mutually financed by the previously stated funding agencies.

  11. Efficient treatment of garbage slurry in methanogenic bioreactor packed by fibrous sponge with high porosity.

    PubMed

    Sasaki, Kengo; Sasaki, Daisuke; Morita, Masahiko; Hirano, Shin-Ichi; Matsumoto, Norio; Ohmura, Naoya; Igarashi, Yasuo

    2010-05-01

    Adding a supporting material to a methanogenic bioreactor treating garbage slurry can improve efficiency of methane production. However, little is known on how characteristics (e.g., porosity and hydrophobicity) of the supporting material affect the bioreactor degrading garbage slurry. We describe the reactor performances and microbial communities in bioreactors containing hydrophilic or hydrophobic sheets, or fibrous hydrophilic or hydrophobic sponges. The porosity affected the efficiency of methane production and solid waste removal more than the hydrophilic or hydrophobic nature of the supporting material. When the terminal restriction fragment length polymorphism technique was used at a lower organic loading rate (OLR), microbial diversities in the suspended fraction were retained on the hydrophobic, but not the hydrophilic, sheets. Moreover, real-time quantitative polymerase chain reaction (PCR) performed at a higher OLR revealed that the excellent performance of reactors containing fibrous sponges with high porosity (98%) was supported by a clear increase in the numbers of methanogens on these sponges, resulting in larger total numbers of methanogens in the reactors. In addition, the bacterial communities in fractions retained on both the hydrophobic and hydrophilic fibrous sponges differed from those in the suspended fraction, thus increasing bacterial diversity in the reactor. Thus, higher porosity of the supporting material improves the bioreactor performance by increasing the amount of methanogens and bacterial diversity; surface hydrophobicity contributes to maintaining the suspended microbial community. PMID:20162271

  12. Assessment of fertilizer potential of the struvite produced from the treatment of methanogenic landfill leachate using low-cost reagents.

    PubMed

    Siciliano, Alessio

    2016-03-01

    Leachates generated in methanogenic landfills contain high strength of ammonium nitrogen which removal is hard to be accomplished by means of conventional techniques. The chemical precipitation of struvite, which is a mineral that could be reused as a slow-release fertilizer, is an effective process in the removal and recovery of NH4 amount of high-concentrated wastewaters. In this paper, a struvite precipitation process using unconventional reagents is proposed for a sustainable recovery of nitrogen content. In particular, seawater bittern, a by-product of marine salt manufacturing, and bone meal, a by-product of the thermal treatment of meat waste, have been used as low-cost sources of magnesium and phosphorus, respectively. The process enables the removal of more than 98 % ammonia load, the recovery about 99 and 95 % of phosphorus and magnesium, respectively, and the production of a precipitate containing struvite crystals. Heavy metals concentrations of produced precipitate were below the threshold values specified by the EC Directive for use of sewage sludges as fertilizers. Specific agronomic tests were conducted to investigate the fertilizing value of precipitate recovered from landfill leachate. The fertilizing effect of struvite deposit in cultivating Spinacia oleracea was compared with that of vegetable soil and commercial fertilizer. The growth of selected vegetable in the pots with struvite precipitate resulted significantly greater in both than those in the control pots and in the pots with the complex fertilizer. Furthermore, the struvite application as fertilizer did not result in more heavy metals in the vegetables respect those from soil and model fertilizer. PMID:26604197

  13. Hydrocarbon activation under sulfate-reducing and methanogenic conditions proceeds by different mechanisms.

    NASA Astrophysics Data System (ADS)

    Head, Ian; Gray, Neil; Aitken, Caroline; Sherry, Angela; Jones, Martin; Larter, Stephen

    2010-05-01

    Microbial degradation of alkanes typically involves their conversion to fatty acids which are then catabolised by beta-oxidation. The critical step in this process is activation of the hydrocarbon. Under oxic conditions this is catalyzed by monooxygenase enzymes with the formation of long chain alcohols. In the absence of oxygen alternative alkane activation mechanisms have been observed or proposed. Fumarate addition to alkanes to form alkyl succinates is considered a central process in anaerobic hydrocarbon degradation. Comparative studies of crude oil degradation under sulphate-reducing and methanogenic conditions revealed distinctive patterns of compound class removal and metabolite formation. Alkyl succinates derived from C7 to C26 n-alkanes and branched chain alkanes were found in abundance in sulfate-reducing systems but these were not detected during methanogenic crude oil degradation. Only one other mechanism of alkane activation has been elucidated to date. This involves addition of carbon derived from bicarbonate/CO2 to C-3 of an alkane chain to form a 2-ethylalkane with subsequent removal of the ethyl group leading to the formation of a fatty acid 1 carbon shorter than the original alkane. 2-ethylalkanes have never been detected as metabolites of anaerobic alkane degradation and were not detected in crude oil-degrading methanogenic systems. Due to the range of alkanes present in crude oil it was not possible to infer the generation of C-odd acids from C-even alkanes which is characteristic of the C-3 carboxylation mechanism. Furthermore genes homologous to alkysuccinate synthetases were not detected in the methanogenic hydrocarbon degrading community by pyrosequencing of total DNA extracted from methanogenic enrichments cultures. beta-oxidation genes were detected and intriguingly, alcohol and aldehyde dehydrogenase genes were present. This offers the possibility that alkane activation in the methanogenic system does not proceed via acid metabolites

  14. How to cultivate Ectocarpus.

    PubMed

    Coelho, Susana M; Scornet, Delphine; Rousvoal, Sylvie; Peters, Nick T; Dartevelle, Laurence; Peters, Akira F; Cock, J Mark

    2012-02-01

    This article describes the standard procedure for growing Ectocarpus in the laboratory. The culture is started with partheno-sporophyte (or sporophyte) filaments because this is the stage that is usually maintained in strain collections. The standard medium is Provasoli-enriched natural seawater (PES), but Ectocarpus can also be grown in artificial seawater, which allows more precise control over the culture conditions. The algae can be cultivated either in plastic Petri dishes or in 10-L bottles with bubbling, if large amounts of biomass are required. Standard growth conditions are 13°C with a 12h/12h d/night cycle and 20 µmol photons m(-2) s(-1) irradiance using daylight-type fluorescent tubes. All manipulations of Ectocarpus cultures should be performed in a clean environment (if possible, under a laminar flow hood). Forceps should be dipped in ethanol and allowed to dry under the hood. PMID:22301662

  15. Investigation into the effect of high concentrations of volatile fatty acids in anaerobic digestion on methanogenic communities.

    PubMed

    Franke-Whittle, Ingrid H; Walter, Andreas; Ebner, Christian; Insam, Heribert

    2014-11-01

    A study was conducted to determine whether differences in the levels of volatile fatty acids (VFAs) in anaerobic digester plants could result in variations in the indigenous methanogenic communities. Two digesters (one operated under mesophilic conditions, the other under thermophilic conditions) were monitored, and sampled at points where VFA levels were high, as well as when VFA levels were low. Physical and chemical parameters were measured, and the methanogenic diversity was screened using the phylogenetic microarray ANAEROCHIP. In addition, real-time PCR was used to quantify the presence of the different methanogenic genera in the sludge samples. Array results indicated that the archaeal communities in the different reactors were stable, and that changes in the VFA levels of the anaerobic digesters did not greatly alter the dominating methanogenic organisms. In contrast, the two digesters were found to harbour different dominating methanogenic communities, which appeared to remain stable over time. Real-time PCR results were inline with those of microarray analysis indicating only minimal changes in methanogen numbers during periods of high VFAs, however, revealed a greater diversity in methanogens than found with the array. PMID:25164858

  16. Shifts in methanogen community structure and function across a coastal marsh transect: effects of exotic Spartina alterniflora invasion

    PubMed Central

    Yuan, Junji; Ding, Weixin; Liu, Deyan; Kang, Hojeong; Xiang, Jian; Lin, Yongxin

    2016-01-01

    Invasion of Spartina alterniflora in coastal areas of China increased methane (CH4) emissions. To elucidate the underlying mechanisms, we measured CH4 production potential, methanogen community structure and biogeochemical factors along a coastal wetland transect comprised of five habitat regions: open water, bare tidal flat, invasive S. alterniflora marsh and native Suaeda salsa and Phragmites australis marshes. CH4 production potential in S. alterniflora marsh was 10 times higher than that in other regions, and it was significantly correlated with soil organic carbon, dissolved organic carbon and trimethylamine concentrations, but was not correlated with acetate or formate concentrations. Although the diversity of methanogens was lowest in S. alterniflora marsh, invasion increased methanogen abundance by 3.48-fold, compared with native S. salsa and P. australis marshes due to increase of facultative Methanosarcinaceae rather than acetotrophic and hydrogenotrophic methanogens. Ordination analyses suggested that trimethylamine was the primary factor regulating shift in methanogen community structure. Addition of trimethylamine increased CH4 production rates by 1255-fold but only by 5.61- and 11.4-fold for acetate and H2/CO2, respectively. S. alterniflora invasion elevated concentration of non-competitive trimethylamine, and shifted methanogen community from acetotrophic to facultative methanogens, which together facilitated increased CH4 production potential. PMID:26728134

  17. Investigation into the effect of high concentrations of volatile fatty acids in anaerobic digestion on methanogenic communities

    PubMed Central

    Franke-Whittle, Ingrid H.; Walter, Andreas; Ebner, Christian; Insam, Heribert

    2014-01-01

    A study was conducted to determine whether differences in the levels of volatile fatty acids (VFAs) in anaerobic digester plants could result in variations in the indigenous methanogenic communities. Two digesters (one operated under mesophilic conditions, the other under thermophilic conditions) were monitored, and sampled at points where VFA levels were high, as well as when VFA levels were low. Physical and chemical parameters were measured, and the methanogenic diversity was screened using the phylogenetic microarray ANAEROCHIP. In addition, real-time PCR was used to quantify the presence of the different methanogenic genera in the sludge samples. Array results indicated that the archaeal communities in the different reactors were stable, and that changes in the VFA levels of the anaerobic digesters did not greatly alter the dominating methanogenic organisms. In contrast, the two digesters were found to harbour different dominating methanogenic communities, which appeared to remain stable over time. Real-time PCR results were inline with those of microarray analysis indicating only minimal changes in methanogen numbers during periods of high VFAs, however, revealed a greater diversity in methanogens than found with the array. PMID:25164858

  18. Shifts in methanogen community structure and function across a coastal marsh transect: effects of exotic Spartina alterniflora invasion

    NASA Astrophysics Data System (ADS)

    Yuan, Junji; Ding, Weixin; Liu, Deyan; Kang, Hojeong; Xiang, Jian; Lin, Yongxin

    2016-01-01

    Invasion of Spartina alterniflora in coastal areas of China increased methane (CH4) emissions. To elucidate the underlying mechanisms, we measured CH4 production potential, methanogen community structure and biogeochemical factors along a coastal wetland transect comprised of five habitat regions: open water, bare tidal flat, invasive S. alterniflora marsh and native Suaeda salsa and Phragmites australis marshes. CH4 production potential in S. alterniflora marsh was 10 times higher than that in other regions, and it was significantly correlated with soil organic carbon, dissolved organic carbon and trimethylamine concentrations, but was not correlated with acetate or formate concentrations. Although the diversity of methanogens was lowest in S. alterniflora marsh, invasion increased methanogen abundance by 3.48-fold, compared with native S. salsa and P. australis marshes due to increase of facultative Methanosarcinaceae rather than acetotrophic and hydrogenotrophic methanogens. Ordination analyses suggested that trimethylamine was the primary factor regulating shift in methanogen community structure. Addition of trimethylamine increased CH4 production rates by 1255-fold but only by 5.61- and 11.4-fold for acetate and H2/CO2, respectively. S. alterniflora invasion elevated concentration of non-competitive trimethylamine, and shifted methanogen community from acetotrophic to facultative methanogens, which together facilitated increased CH4 production potential.

  19. Hybrid origins of cultivated potatoes

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Wild and cultivated potatoes, Solanum section Petota, is taxonomically difficult, partly because of interspecific hybridization at both the diploid and polyploid levels. The taxonomy of cultivated potatoes is particularly controversial. With DNA sequence data of the GBSSI (waxy) gene we here infer r...

  20. Micrometeorological principles of protected cultivation

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Protected cultivation is a broad term commonly used among producers of specialty crops. Techniques can range from complex fixed structures to field site selection, to straightforward cultural practices in the field. This introduction to the ASHS workshop "Protected cultivation for fruit crops" consi...

  1. Starting from grape cultivation.

    PubMed

    Yoshida, A

    1992-06-01

    Rapid population growth can only be stopped by lowering the fertility rate. The UNFPA recommends improving the employment opportunities for women as the single best way of achieving this reduction. An example of this phenomenon is the grape cultivation in the Nordeste (Northeastern) region of Brazil. This area is the poorest part of Brazil and has the highest proportion of indigent people. These people have been deforesting the Amazon in search of a better life. What they have done is sterilize the land and turned a tropical rain forest into a desert. In an effort to reverse this trend, grape cultivation has been introduced in an area called Petrolina. The area is very dry with less than 500 mm of precipitation annually. They do have access to a 5000 square kilometer artificial lake (the largest in the world) and the 3rd largest river in Brazil (the Sao Francisco). In an effort to avoid using agricultural medicines, the vines are fertilized with organic matter created on the farm and little or no pesticides are used since pests do not live in such an arid region. It has taken 20 years of trial and error, but the quality of the grapes is now very high and is competitive on the world market. Because of climate and location, harvesting is done year round which increases the productivity of the land. The farm managers have found that married women make the best workers and have the highest level of productivity. Age at 1st marriage averages 24-25, compared with 15-16 for unemployed women in the same area. The fertility rate averages 50% of that for unemployed women in the same area. Agricultural development offers the best opportunity for the women of developing countries. It can pay a high wage, reduce fertility, and replant desert areas. PMID:12285551

  2. Seasonal Changes in Methanogenesis and Methanogenic Community in Three Peatlands, New York State

    PubMed Central

    Sun, Christine L.; Brauer, Suzanna L.; Cadillo-Quiroz, Hinsby; Zinder, Stephen H.; Yavitt, Joseph B.

    2012-01-01

    Fluctuating environmental conditions can promote diversity and control dominance in community composition. In addition to seasonal temperature and moisture changes, seasonal supply of metabolic substrates selects populations temporally. Here we demonstrate cascading effects in the supply of metabolic substrates on methanogenesis and community composition of anaerobic methanogenic archaea in three contrasting peatlands in upstate New York. Fresh samples of peat soils, collected about every 3 months for 20 months and incubated at 22 ± 2°C regardless of the in situ temperature, exhibited potential rates of methane (CH4) production of 0.02–0.2 mmol L−1 day−1 [380–3800 nmol g−1 (dry) day−1). The addition of acetate stimulated rates of CH4 production in a fen peatland soil, whereas addition of hydrogen (H2), and simultaneous inhibition of H2-consuming acetogenic bacteria with rifampicin, stimulated CH4 production in two acidic bog soils, especially, in autumn and winter. The methanogenic community structure was characterized using T-RFLP analyses of SSU rRNA genes. The E2 group of methanogens (Methanoregulaceae) dominated in the two acidic bog peatlands with relatively greater abundance in winter. In the fen peatland, the E1 group (Methanoregulaceae) and members of the Methanosaetaceae were co-dominant, with E1 having a high relative abundance in spring. Change in relative abundance profiles among methanogenic groups in response to added metabolic substrates was as predicted. The acetate-amendment increased abundance of Methanosarcinaceae, and H2-amendment enhanced abundance of E2 group in all peat soils studied, respectively. Additionally, addition of acetate increased abundance of Methanosaetaceae only in the bog soils. Variation in the supply of metabolic substrates helps explain the moderate diversity of methanogens in peatlands. PMID:22408638

  3. Roles of thermophilic thiosulfate-reducing bacteria and methanogenic archaea in the biocorrosion of oil pipelines

    PubMed Central

    Liang, Renxing; Grizzle, Robert S.; Duncan, Kathleen E.; McInerney, Michael J.; Suflita, Joseph M.

    2014-01-01

    Thermophilic sulfide-producing microorganisms from an oil pipeline network were enumerated with different sulfur oxyanions as electron acceptors at 55°C. Most-probable number (MPN) analysis showed that thiosulfate-reducing bacteria were the most numerous sulfidogenic microorganisms in pipeline inspection gauge (PIG) scrapings. Thiosulfate-reducing and methanogenic enrichments were obtained from the MPN cultures that were able to use yeast extract as the electron donor. Molecular analysis revealed that both enrichments harbored the same dominant bacterium, which belonged to the genus Anaerobaculum. The dominant archaeon in the methanogenic enrichment was affiliated with the genus Methanothermobacter. With yeast extract as the electron donor, the general corrosion rate by the thiosulfate-reducing enrichment (8.43 ± 1.40 milli-inch per year, abbreviated as mpy) was about 5.5 times greater than the abiotic control (1.49 ± 0.15 mpy), while the comparable measures for the methanogenic culture were 2.03 ± 0.49 mpy and 0.62 ± 0.07 mpy, respectively. Total iron analysis in the cultures largely accounted for the mass loss of iron measured in the weight loss determinations. Profilometry analysis of polished steel coupons incubated in the presence of the thiosulfate-reducing enrichment revealed 59 pits over an area of 71.16 mm2, while only 6 pits were evident in the corresponding methanogenic incubations. The results show the importance of thiosulfate-utilizing, sulfide-producing fermentative bacteria such as Anaerobaculum sp. in the corrosion of carbon steel, but also suggest that Anaerobaculum sp. are of far less concern when growing syntrophically with methanogens. PMID:24639674

  4. The role of methanogens in acetic acid production under different salinity conditions.

    PubMed

    Xiao, Keke; Guo, Chenghong; Maspolim, Yogananda; Zhou, Yan; Ng, Wun Jern

    2016-10-01

    In this study, a fed-batch acidogenic reactor was operated at a 3 d hydraulic retention time (HRT) and fed with alkaline pre-treated sludge to investigate salinity effects on methanogens' abundance, activities and their consumption of produced acetic acid (HAc) and total volatile fatty acids (VFAs). The salinity concentration was increased step-wise by adding sodium chloride. At 3‰ (parts per thousand) salinity, the average produced volatile fatty acids (VFAs) concentration was 2410.16 ± 637.62 mg COD L(-1) and 2.70 ± 0.36 L methane was produced daily in the acidogenic reactor. Further batch tests indicated methanogens showed a HAc degradation rate of 3.81 mg COD g(-1) VSS h(-1) at initial HAc concentration of 1150 mg COD L(-1), and showed tolerance up to 16‰ salinity (3.76 g Na(+) L(-1)) as indicated by a constant HAc degradation rate. The microbiological study indicated this can be related to the predominance of acetate-utilizing Methanosarcinaceae and Methanomicrobiales in the reactor. However, with salinity increased to 20‰ and 40‰, increases in VFAs and HAc production and decreases in methane production, methanogens population, acidogenic bacteria population and acidification extent were observed. This study demonstrated presence of acetate-utilizing methanogens in an acidogenic reactor and their high tolerance to salinity, as well as their negative impacts on net VFAs production. The results would suggest the presence of methanogens in the acidogenic reactor should not be ignored and the recovery of methane from the acidogenic reactor needs to be considered to avoid carbon loss. PMID:27421101

  5. Methanogenic Population and CH4 Production in Swedish Dairy Cows Fed Different Levels of Forage

    PubMed Central

    Schnürer, A.; Arthurson, V.; Bertilsson, J.

    2012-01-01

    Methanogenic community structure, methane production (CH4), and volatile fatty acid (VFA) profiles were investigated in Swedish dairy cows fed a diet with a forage/concentrate ratio of 500/500 or 900/100 g/kg of dry matter (DM) of total DM intake (DMI). The rumen methanogenic population was evaluated using terminal restriction fragment length polymorphism (T-RFLP) analysis, 16S rRNA gene libraries, and quantitative real-time PCR (qRT-PCR). Mean CH4 yields did not differ (P > 0.05) between diets, being 16.9 and 20.2 g/kg DMI for the 500/500 and 900/100 diets, respectively. The T-RFLP analysis revealed that populations differed between individual cows and that each individual population responded differently to the diets. The 16S rRNA gene libraries revealed that Methanobrevibacter spp. dominated for both diets. CH4 production was positively correlated with a dominance of sequences representing T-RFs related to Methanobrevibacter thaueri, Methanobrevibacter millerae, and Methanobrevibacter smithii relative to Methanobrevibacter ruminantium and Methanobrevibacter olleyae. Total numbers of methanogens and total numbers of Methanobacteriales were significantly higher with the 500/500 diet (P < 0.0004 and P < 0.002, respectively). However, no relationship was found between CH4 production and total number of methanogens. No differences were seen in total VFA, propionic acid, or acetic acid contents, but the molar proportion of butyric acid in the rumen was higher for the 500/500 diet than for the 900/100 diet (P < 0.05). Interestingly, the results also revealed that a division of the identified methanogenic species into two groups, suggested in the work of King et al. (E. E. King, R. P. Smith, B. St-Pierre, and A. D. G. Wright, Appl. Environ. Microbiol. 77:5682–5687, 2011), increased the understanding of the variation in CH4 production between different cows. PMID:22752163

  6. Laser spectroscopic real time measurements of methanogenic activity under simulated Martian subsurface analog conditions

    NASA Astrophysics Data System (ADS)

    Schirmack, Janosch; Böhm, Michael; Brauer, Chris; Löhmannsröben, Hans-Gerd; de Vera, Jean-Pierre; Möhlmann, Diedrich; Wagner, Dirk

    2014-08-01

    On Earth, chemolithoautothrophic and anaerobic microorganisms such as methanogenic archaea are regarded as model organisms for possible subsurface life on Mars. For this reason, the methanogenic strain Methanosarcina soligelidi (formerly called Methanosarcina spec. SMA-21), isolated from permafrost-affected soil in northeast Siberia, has been tested under Martian thermo-physical conditions. In previous studies under simulated Martian conditions, high survival rates of these microorganisms were observed. In our study we present a method to measure methane production as a first attempt to study metabolic activity of methanogenic archaea during simulated conditions approaching conditions of Mars-like environments. To determine methanogenic activity, a measurement technique which is capable to measure the produced methane concentration with high precision and with high temporal resolution is needed. Although there are several methods to detect methane, only a few fulfill all the needed requirements to work within simulated extraterrestrial environments. We have chosen laser spectroscopy, which is a non-destructive technique that measures the methane concentration without sample taking and also can be run continuously. In our simulation, we detected methane production at temperatures down to -5 °C, which would be found on Mars either temporarily in the shallow subsurface or continually in the deep subsurface. The pressure of 50 kPa which we used in our experiments, corresponds to the expected pressure in the Martian near subsurface. Our new device proved to be fully functional and the results indicate that the possible existence of methanogenic archaea in Martian subsurface habitats cannot be ruled out.

  7. Trace Elements Affect Methanogenic Activity and Diversity in Enrichments from Subsurface Coal Bed Produced Water

    PubMed Central

    Ünal, Burcu; Perry, Verlin Ryan; Sheth, Mili; Gomez-Alvarez, Vicente; Chin, Kuk-Jeong; Nüsslein, Klaus

    2012-01-01

    Microbial methane from coal beds accounts for a significant and growing percentage of natural gas worldwide. Our knowledge of physical and geochemical factors regulating methanogenesis is still in its infancy. We hypothesized that in these closed systems, trace elements (as micronutrients) are a limiting factor for methanogenic growth and activity. Trace elements are essential components of enzymes or cofactors of metabolic pathways associated with methanogenesis. This study examined the effects of eight trace elements (iron, nickel, cobalt, molybdenum, zinc, manganese, boron, and copper) on methane production, on mcrA transcript levels, and on methanogenic community structure in enrichment cultures obtained from coal bed methane (CBM) well produced water samples from the Powder River Basin, Wyoming. Methane production was shown to be limited both by a lack of additional trace elements as well as by the addition of an overly concentrated trace element mixture. Addition of trace elements at concentrations optimized for standard media enhanced methane production by 37%. After 7 days of incubation, the levels of mcrA transcripts in enrichment cultures with trace element amendment were much higher than in cultures without amendment. Transcript levels of mcrA correlated positively with elevated rates of methane production in supplemented enrichments (R2 = 0.95). Metabolically active methanogens, identified by clone sequences of mcrA mRNA retrieved from enrichment cultures, were closely related to Methanobacterium subterraneum and Methanobacterium formicicum. Enrichment cultures were dominated by M. subterraneum and had slightly higher predicted methanogenic richness, but less diversity than enrichment cultures without amendments. These results suggest that varying concentrations of trace elements in produced water from different subsurface coal wells may cause changing levels of CBM production and alter the composition of the active methanogenic community. PMID

  8. Interspecies distances between propionic acid degraders and methanogens in syntrophic consortia for optimal hydrogen transfer.

    PubMed

    Felchner-Zwirello, Monika; Winter, Josef; Gallert, Claudia

    2013-10-01

    A mixed culture from an anaerobic biowaste digester was enriched on propionate and used to investigate interspecies hydrogen transfer in dependence of spatial distances between propionate degraders and methanogens. From 20.3 mM propionate, 20.8 mM acetate and 15.5 mM methane were formed. Maximum specific propionate oxidation and methane formation rates were 49 and 23 mmol mg(-1) day(-1), respectively. Propionate oxidation was inhibited by only 20 mM acetate by about 50 %. Intermediate formate formation during inhibited methanogensis was observed. The spatial distribution and the biovolume fraction of propionate degraders and of methanogens in relation to the total population during aggregate formation were determined. Measurements of interbacterial distances were conducted with fluorescence in situ hybridization by application of group-specific 16S rRNA-targeted probes and 3D image analyses. With increasing incubation time, floc formation and growth up to 54 μm were observed. Propionate degraders and methanogens were distributed randomly in the flocs. The methanogenic biovolume fraction was high at the beginning and remained constant over 42 days, whereas the fraction of propionate degraders increased with time during propionate feeding. Interbacterial distances between propionate degraders and methanogens decreased with time from 5.30 to 0.29 μm, causing an increase of the maximum possible hydrogen flux from 1.1 to 10.3 nmol ml(-1) min(-1). The maximum possible hydrogen flux was always higher than the hydrogen formation and consumption rate, indicating that reducing the interspecies distance by aggregation is advantageous in complex ecosystems. PMID:23233207

  9. Roles of thermophilic thiosulfate-reducing bacteria and methanogenic archaea in the biocorrosion of oil pipelines.

    PubMed

    Liang, Renxing; Grizzle, Robert S; Duncan, Kathleen E; McInerney, Michael J; Suflita, Joseph M

    2014-01-01

    Thermophilic sulfide-producing microorganisms from an oil pipeline network were enumerated with different sulfur oxyanions as electron acceptors at 55°C. Most-probable number (MPN) analysis showed that thiosulfate-reducing bacteria were the most numerous sulfidogenic microorganisms in pipeline inspection gauge (PIG) scrapings. Thiosulfate-reducing and methanogenic enrichments were obtained from the MPN cultures that were able to use yeast extract as the electron donor. Molecular analysis revealed that both enrichments harbored the same dominant bacterium, which belonged to the genus Anaerobaculum. The dominant archaeon in the methanogenic enrichment was affiliated with the genus Methanothermobacter. With yeast extract as the electron donor, the general corrosion rate by the thiosulfate-reducing enrichment (8.43 ± 1.40 milli-inch per year, abbreviated as mpy) was about 5.5 times greater than the abiotic control (1.49 ± 0.15 mpy), while the comparable measures for the methanogenic culture were 2.03 ± 0.49 mpy and 0.62 ± 0.07 mpy, respectively. Total iron analysis in the cultures largely accounted for the mass loss of iron measured in the weight loss determinations. Profilometry analysis of polished steel coupons incubated in the presence of the thiosulfate-reducing enrichment revealed 59 pits over an area of 71.16 mm(2), while only 6 pits were evident in the corresponding methanogenic incubations. The results show the importance of thiosulfate-utilizing, sulfide-producing fermentative bacteria such as Anaerobaculum sp. in the corrosion of carbon steel, but also suggest that Anaerobaculum sp. are of far less concern when growing syntrophically with methanogens. PMID:24639674

  10. Methanogenic activities in anaerobic membrane bioreactors (AnMBR) treating synthetic municipal wastewater.

    PubMed

    Ho, Jaeho; Sung, Shihwu

    2010-04-01

    Two laboratory-scale anaerobic membrane bioreactors, AnMBR 1 and AnMBR 2, were run in parallel at 25 and 15 degrees C, respectively. Total chemical oxygen demand (COD) removal efficiency was more than 95% and 85% for AnMBR 1 and 2, respectively. The COD removal of AnMBR 1 was mostly carried out biologically. However, the physical removal on the membrane surface compensated for the decreased biological removal rate in AnMBR 2. The membrane in AnMBR systems is likely not only to retain all biomass in the reactor, but also complement decreased biological removal efficiency at low temperature by rejecting soluble organics. Specific methanogenic activity (SMA) test was used to investigate the methanogenic activity profiles of suspended and attached sludge in AnMBRs treating synthetic municipal wastewater at 25 and 15 degrees C. The methanogenic activity was 51.8 ml CH(4)/g VSSd on day 1 and eventually increased 27% and reached 65.7 ml CH(4)/g VSSd on day 75 for AnMBR 1. However, the methanogenic activity of AnMBR 2 sludge was lower than that of AnMBR 1. The microbial activity of suspended sludge continuously increased, while that of attached sludge gradually decreased in this study. The methanogenic activity of attached sludge was far lower than that of suspended sludge. The role of attached sludge on the membrane in AnMBRs as a biofilm for biological organic removal was minimal compared to suspended sludge. PMID:20022745

  11. Use of a Hierarchical Oligonucleotide Primer Extension Approach for Multiplexed Relative Abundance Analysis of Methanogens in Anaerobic Digestion Systems

    PubMed Central

    Chuang, Hui-Ping; Hsu, Mao-Hsuan; Chen, Wei-Yu

    2013-01-01

    In this study, we established a rapid multiplex method to detect the relative abundances of amplified 16S rRNA genes from known cultivatable methanogens at hierarchical specificities in anaerobic digestion systems treating industrial wastewater and sewage sludge. The method was based on the hierarchical oligonucleotide primer extension (HOPE) technique and combined with a set of 27 primers designed to target the total archaeal populations and methanogens from 22 genera within 4 taxonomic orders. After optimization for their specificities and detection sensitivity under the conditions of multiple single-nucleotide primer extension reactions, the HOPE approach was applied to analyze the methanogens in 19 consortium samples from 7 anaerobic treatment systems (i.e., 513 reactions). Among the samples, the methanogen populations detected with order-level primers accounted for >77.2% of the PCR-amplified 16S rRNA genes detected using an Archaea-specific primer. The archaeal communities typically consisted of 2 to 7 known methanogen genera within the Methanobacteriales, Methanomicrobiales, and Methanosarcinales and displayed population dynamic and spatial distributions in anaerobic reactor operations. Principal component analysis of the HOPE data further showed that the methanogen communities could be clustered into 3 distinctive groups, in accordance with the distribution of the Methanosaeta, Methanolinea, and Methanomethylovorans, respectively. This finding suggested that in addition to acetotrophic and hydrogenotrophic methanogens, the methylotrophic methanogens might play a key role in the anaerobic treatment of industrial wastewater. Overall, the results demonstrated that the HOPE approach is a specific, rapid, and multiplexing platform to determine the relative abundances of targeted methanogens in PCR-amplified 16S rRNA gene products. PMID:24077716

  12. Study on two methylotrophic and halophilic methanogens, Methanosarcina siciliae HI350 and Methanolobus taylorii GS-16

    SciTech Connect

    Ni, S.

    1994-01-01

    Strain HI350, similar to Methanolobus siciliae T4/M[sup T] (T = type strain) morphologically and physiologically, was isolated from an oil well in the Gulf of Mexico. Catabolic substrates included methanol, trimethylamine, dimethyl sulfide, and methane thiol, but not H[sub 2]-CO[sub 2], formate, or acetate. Growth was fastest in the presence of 0.4 to 0.6 M Na[sup +], in the presence of 60 to 200 mM Mg[sup 2+], at pH 6.5 to 6.8, and at 40[degrees]C. Methanolobus siciliae T4/M[sup T] was closely related to Methanosarcina. Transfer of Methanolobus siciliae T4/M[sup T] to the genus Methanosarcina as Methanosarcina siciliae is proposed with strain HI350 as its reference strain. Degradation of dimethyl sulfide or methane thiol by strain HI350 was complete, and stoichiometric quantities of methane and hydrogen sulfide were formed. Studies of cell-free extracts suggested that enzymes for degradation of dimethyl sulfide and methane thiol were inducible, whereas those for the degradation of methanol or trimethylamine were constitutive. Methanolobus taylorii GS-16, a moderately halophilic and alkcaliphilic methanogen, grows over a wide pH range. The key observation indicative of the involvement of K[sup +] transport in cytosolic acidification was that valinomycin (0.8 [mu]M), a K[sup +] uniporter, inhibited the growth of strain GS-16 only at alkaline pH. Experiments with resting cells indicated that, at alkaline pH, valinomycin uncoupled catabolic reactions from ATP synthesis. Thus, a K[sup +]/H[sup +] antiporter was proposed to account for the K[sup +] extrusion and the uncoupling effect of valinomycin at alkaline pH.

  13. Propioniciclava tarda gen. nov., sp. nov., isolated from a methanogenic reactor treating waste from cattle farms.

    PubMed

    Sugawara, Yoshimichi; Ueki, Atsuko; Abe, Kunihiro; Kaku, Nobuo; Watanabe, Kazuya; Ueki, Katsuji

    2011-09-01

    Two facultatively anaerobic bacterial strains, designated WR061(T) and WR054, were isolated from rice-straw residue in a methanogenic reactor treating waste from cattle farms in Japan. The two strains were phylogenetically positioned close to one another and had almost the same phenotypic properties. Cells were Gram-reaction-positive, non-motile, non-spore-forming, irregular rods. Cobalamin (vitamin B₁₂) was required for growth. The strains utilized various carbohydrates, including hexoses and disaccharides, and produced acetate and propionate from these carbohydrates. Pentoses and polysaccharides were not utilized. They grew at 20-37 °C (optimum 35 °C) and pH 5.3-8.0 (optimum pH 6.8-7.5). Catalase and nitrate-reducing activities were detected. Aesculin was hydrolysed. The major cellular fatty acids were anteiso-C₁₅:₀ and C₁₅:₀ DMA, the major respiratory quinone was menaquinone MK-9(H₄) and the genomic DNA G+C content was 69.3-69.5  mol%. The diagnostic diamino acid in the peptidoglycan was meso-diaminopimelic acid. Phylogenetic analysis based on 16S rRNA gene sequences placed the strains in the phylum Actinobacteria. Both strains were remotely related to the species in the family Propionibacteriaceae and Propionibacterium propionicum JCM 5830(T) was the most closely related type strain with a sequence similarity of 91.6 %. Based on phylogenetic, physiological and chemotaxonomic analyses, the two novel strains together represent a novel species of a new genus, for which the name Propioniciclava tarda gen. nov., sp. nov. is proposed. The type strain is WR061(T) ( = JCM 15804(T)  = DSM 22130(T)). PMID:20971831

  14. Bacteroides paurosaccharolyticus sp. nov., isolated from a methanogenic reactor treating waste from cattle farms.

    PubMed

    Ueki, Atsuko; Abe, Kunihiro; Ohtaki, Yoshimi; Kaku, Nobuo; Watanabe, Kazuya; Ueki, Katsuji

    2011-02-01

    A strictly anaerobic bacterial strain (WK042(T)) was isolated from rice-straw residue in a methanogenic reactor treating waste from cattle farms in Japan. Cells were Gram-staining-negative, non-motile, non-spore-forming rods. Growth was stimulated well by haemin, and was enhanced by cobalamin (vitamin B(12)). Strain WK042(T) utilized arabinose, xylose, glucose, mannose and aesculin as preferred substrates. Maltose, dextrin, glycogen, starch and pectin were also utilized, although growth on these substrates was much slower. The strain produced acetate, propionate and succinate from these saccharides. The strain was slightly alkaliphilic, with optimum growth at pH 7.7. The temperature range for growth was 10-40 °C, the optimum being 35 °C. The strain was sensitive to bile. The major cellular fatty acids were anteiso-C(15 : 0), iso-C(17 : 0) 3-OH and C(15 : 0). Menaquinone 11 (MK-11) was the major respiratory quinone and the genomic DNA G+C content was 41.0 mol%. Phylogenetic analysis based on 16S rRNA gene sequences placed the strain in the phylum Bacteroidetes. Strain WK042(T) was related distantly to the type strains of species in the cluster including Bacteroides massiliensis, Bacteroides vulgatus and Bacteroides dorei (91-92 % 16S rRNA gene sequence similarity). Based on data from the present phylogenetic, physiological and chemotaxonomic analyses, strain WK042(T) is considered to represent a novel species of the genus Bacteroides, for which the name Bacteroides paurosaccharolyticus sp. nov. is proposed. The type strain is WK042(T) (=JCM 15092(T) =DSM 21004(T)). PMID:20348319

  15. Methanogenic biodegradation of creosote-derived contaminants in natural and simulated ground water ecosystems

    SciTech Connect

    Godsy, E.M.

    1993-12-31

    Wastes from an abandoned wood preserving plant in Pensacola, Florida have contaminated the near surface sand-and-gravel aquifer with creosote-derived compounds and pentachlorophenol. The contaminated ground water is enriched in organic fatty acids, benzoic acid, phenol, 2-, 3-, 4-methylphenol, indole, oxindole, quinoline, isoquinoline, 1(2H)-quinolinone, 2(1H)-isoquinolinone, benzothiophene, benzofuran, naphthalene, and indene. Evidence is presented that the methanogenic degradation of the compounds listed above and concomitant microbial growth in batch microcosms derived from contaminated aquifer material can be described using Monod kinetics. The K{sub s} values obtained for the biodegradation of the phenolic compounds in this study are much lower than published values, indicating that the aquifer microorganisms may have developed enzyme systems and transport mechanisms that are adapted to low nutrient conditions. The values for k{sub d} are much less than {mu}{sub max}, and can be neglected in the microcosm studies. The low Y values, approximately an order of magnitude lower than theoretical values, and the low numbers of microorganisms in the aquifer derived microcosms suggest that these organisms may use some unique strategies to survive in the subsurface environment. Studies of the degradation pathways of the homocyclic and heterocyclic aromatic compounds on the basis of intermediate compounds have revealed that the process consists of both a major and a minor pathway. The first transformation step of the major pathway consists of oxidation and cleavage of the heterocyclic ring. After cleavage of this ring, the substituent side chains and the remaining homocyclic ring are subjected to various reactions including oxidation, decarboxylation, desulfurylation or ammonification, and O-methylation. These reactions are followed by the reduction of the homocyclic ring, cleavage of this ring, {beta}-oxidation, and mineralization.

  16. Enzymological and genetic studies of one-carbon reactions in methanogenic bacteria. Annual report, January 1987-December 1987

    SciTech Connect

    Ferry, J.G.

    1988-01-01

    This project is aimed at determining the biochemical mechanisms of one-carbon reactions and the genetic-control mechanisms of methanogenesis, an important step in the conversion of biomass to methane. Two enzymes (acetate kinase and a ferredoxin) were purified from methanogenic bacteria. These results extend the understanding of how methanogens regulate metabolism and identify a control point that could be exploited to improve the rates of acetate conversion to methane. A plasmid DNA from methanogens was purified and characterized which could potentially provide a tool for genetic engineering to provide new methods to further study steps in the pathway.

  17. Methanogenic activity in plankton samples and fish intestines A mechanism for in situ methanogenesis in oceanic surface waters

    USGS Publications Warehouse

    Oremland, Ronald S.

    1979-01-01

    When plankton samples were incubated anaerobically with a cysteine-sulfide reducing agent, pronounced methane evolution occurred. This activity was inhibited by air, CHCl3, C2H2, and 2-bromoethanesulfonic acid. Adding [14C]CO32− resulted in accumulation of [14C]CH4. Portions of the digestive tracts of three fishes were incubated in methanogenic media, and two of the samples showed the presence of methanogenic bacteria.

  18. Field and laboratory studies of methane oxidation in an anoxic marine sediment: Evidence for a methanogen-sulfate reducer consortium

    SciTech Connect

    Hoehler, T.M.; Alperin, M.J.; Albert, D.B.

    1994-12-01

    Field and laboratory studies of anoxic sediments from Cape Lookout Bight, North Carolina, suggest that anaerobic methane oxidation is mediated by a consortium of methanogenic and sulfate-reducing bacteria. A seasonal survey of methane oxidation and CO{sub 2} reduction rates indicates that methane production was confined to sulfate-depleted sediments at all times of year, while methane oxidation occurred in two modes. In the summer, methane oxidation was confined to sulfate-depleted sediments and occurred at rates lower than those of CO{sub 2} reduction. In the winter, net methane oxidation occurred in an interval at the base of the sulfate-containing zone. Sediment incubation experiments suggest both methanogens and sulfate reducers were responsible for the observed methane oxidation. In one incubation experiment both modes of oxidation were partially inhibited by 2-bromoethanesulfonic acid (a specific inhibitor of methanogens). This evidence, along with the apparent confinement of methane oxidation to sulfate-depleted sediments in the summer, indicates that methanogenic bacteria are involved in methane oxidation. In a second incubation experiment, net methane oxidation was induced by adding sulfate to homogenized methanogenic sediments, suggesting that sulfate reducers also a play a role in the process. We hypothesize that methanogens oxidize methane and produce hydrogen via a reversal of CO{sub 2} reduction. The hydrogen is efficiently removed and maintained at low concentrations by sulfate reducers. Pore water H{sub 2} concentrations in the sediment incubation experiments (while net methane oxidation was occurring) were low enough that methanogenic bacteria could derive sufficient energy for growth from the oxidation of methane. The methanogen-sulfate reducer consortium may also be a feasible mechanism for previously documented anaerobic methane oxidation in both freshwater and marine environments. 63 refs., 6 refs.

  19. Confocal Raman microspectroscopy reveals a convergence of the chemical composition in methanogenic archaea from a Siberian permafrost-affected soil.

    PubMed

    Serrano, Paloma; Hermelink, Antje; Lasch, Peter; de Vera, Jean-Pierre; König, Nicole; Burckhardt, Oliver; Wagner, Dirk

    2015-12-01

    Methanogenic archaea are widespread anaerobic microorganisms responsible for the production of biogenic methane. Several new species of psychrotolerant methanogenic archaea were recently isolated from a permafrost-affected soil in the Lena Delta (Siberia, Russia), showing an exceptional resistance against desiccation, osmotic stress, low temperatures, starvation, UV and ionizing radiation when compared to methanogens from non-permafrost environments. To gain a deeper insight into the differences observed in their resistance, we described the chemical composition of methanogenic strains from permafrost and non-permafrost environments using confocal Raman microspectroscopy (CRM). CRM is a powerful tool for microbial identification and provides fingerprint-like information about the chemical composition of the cells. Our results show that the chemical composition of methanogens from permafrost-affected soils presents a high homology and is remarkably different from strains inhabiting non-permafrost environments. In addition, we performed a phylogenetic reconstruction of the studied strains based on the functional gene mcrA to prove the different evolutionary relationship of the permafrost strains. We conclude that the permafrost methanogenic strains show a convergent chemical composition regardless of their genotype. This fact is likely to be the consequence of a complex adaptive process to the Siberian permafrost environment and might be the reason underlying their resistant nature. PMID:26499486

  20. Abundance and potential metabolic activity of methanogens in well-aerated forest and grassland soils of an alpine region.

    PubMed

    Hofmann, Katrin; Praeg, Nadine; Mutschlechner, Mira; Wagner, Andreas O; Illmer, Paul

    2016-02-01

    Although methanogens were recently discovered to occur in aerated soils, alpine regions have not been extensively studied for their presence so far. Here, the abundance of archaea and the methanogenic guilds Methanosarcinales, Methanococcales, Methanobacteriales, Methanomicrobiales and Methanocella spp. was studied at 16 coniferous forest and 14 grassland sites located at the montane and subalpine belts of the Northern Limestone Alps (calcareous) and the Austrian Central Alps (siliceous) using quantitative real-time PCR. Abundance of archaea, methanogens and the methanogenic potentials were significantly higher in grasslands than in forests. Furthermore, methanogenic potentials of calcareous soils were higher due to pH. Methanococcales, Methanomicrobiales and Methanocella spp. were detected in all collected samples, which indicates that they are autochthonous, while Methanobacteriales were absent from 4 out of 16 forest soils. Methanosarcinales were absent from 10 out of 16 forest soils and 2 out of 14 grassland soils. Nevertheless, together with Methanococcales they represented the majority of the 16S rRNA gene copies quantified from the grassland soils. Contrarily, forest soils were clearly dominated by Methanococcales. Our results indicate a higher diversity of methanogens in well-aerated soils than previously believed and that pH mainly influences their abundances and activities. PMID:26712349

  1. Divergent responses of methanogenic archaeal communities in two rice cultivars to elevated ground-level O3.

    PubMed

    Zhang, Jianwei; Tang, Haoye; Zhu, Jianguo; Lin, Xiangui; Feng, Youzhi

    2016-06-01

    Inhibitive effect of elevated ground-level ozone (O3) on paddy methane (CH4) emission varies with rice cultivars. However, little information is available on its microbial mechanism. For this purpose, the responses of methane-metabolizing microorganisms, methanogenic archaea and methanotrophic bacteria to O3 pollution were investigated in the O3-tolerant (YD6) and the O3-sensitive (IIY084) cultivars at two rice growth stages in Free Air Concentration Elevation of O3 (O3-FACE) system of China. It was found that O3 pollution didn't change the abundances of Type I and Type II methanotrophic bacteria at two rice stages. For methanogenic archaea, their abundances in both cultivars were decreased by O3 pollution at the tillering stage. Furthermore, a greater negative influence on methanogenic archaeal community was observed on IIY084 than on YD6: at tillering stage, the alpha diversity indices of methanogenic archaeal community in IIY084 was decreased to a greater extent than in YD6; IIY084 shifted methanogenic archaeal community composition and decreased the abundances and the diversities of Methanosarcinaceae and Methanosaetaceae as well as the abundance of Methanomicrobiales, while the diversity of Methanocellaceae were increased in YD6. These findings indicate that the variations in the responses of paddy CH4 emission to O3 pollution between cultivars could result from the divergent responses of their methanogenic archaea. PMID:26895536

  2. Methanogenic diversity and activity in hypersaline sediments of the centre of the Napoli mud volcano, Eastern Mediterranean Sea.

    PubMed

    Lazar, Cassandre Sara; Parkes, R John; Cragg, Barry A; L'Haridon, Stéphane; Toffin, Laurent

    2011-08-01

    Submarine mud volcanoes are a significant source of methane to the atmosphere. The Napoli mud volcano, situated in the brine-impacted Olimpi Area of the Eastern Mediterranean Sea, emits mainly biogenic methane particularly at the centre of the mud volcano. Temperature gradients support the suggestion that Napoli is a cold mud volcano with moderate fluid flow rates. Biogeochemical and molecular genetic analyses were carried out to assess the methanogenic activity rates, pathways and diversity in the hypersaline sediments of the centre of the Napoli mud volcano. Methylotrophic methanogenesis was the only significant methanogenic pathway in the shallow sediments (0-40 cm) but was also measured throughout the sediment core, confirming that methylotrophic methanogens could be well adapted to hypersaline environments. Hydrogenotrophic methanogenesis was the dominant pathway below 50 cm; however, low rates of acetoclastic methanogenesis were also present, even in sediment layers with the highest salinity, showing that these methanogens can thrive in this extreme environment. PCR-DGGE and methyl coenzyme M reductase gene libraries detected sequences affiliated with anaerobic methanotrophs (mainly ANME-1) as well as Methanococcoides methanogens. Results show that the hypersaline conditions in the centre of the Napoli mud volcano influence active biogenic methane fluxes and methanogenic/methylotrophic diversity. PMID:21382146

  3. Molecular biology and physiology of methanogenic archaebacteria. Annual report, July 1988-June 1989

    SciTech Connect

    Nagle, D.P.; McCarthy, D.; Tanner, R.S.

    1989-06-27

    Methane-producing archaebacteria are worthy of their novel biology and potential in anaerobic bioprocessing. This work continues to study the biochemistry, genetics, and molecular biology of the thermophilic autotroph Methanobacterium thermoautotrophicum. DNA from antimetabolite-resistant mutant strains was used to transform sensitive recipient cells to resistance, and DNA was cloned into Escherichia coli plasmids. This DNA will be mutated with transposons in the E. coli host, then isolated and used to transform methanogen cells to selectable mutant phenotypes. Mutant strains resistant to purine analogs were used to determine that wild type cells of M. thermoautotrophicum possess an almost complete set of enzymes for uptake, activation, and interconversion of purine bases and nucleosides. These mutants and the information about the pathways will be the basis for generating a genetic map. Metabolic studies of a unique formate auxotroph revealed a new role for this one carbon compound in the anabolic metabolism of this methanogen.

  4. Clogging of landfill tyre and aggregate drainage layers by methanogenic leachate and implications for practice.

    PubMed

    Beaven, R P; Hudson, A P; Knox, K; Powrie, W; Robinson, J P

    2013-02-01

    This paper reports the results of pilot scale tests carried out to investigate the clogging of shredded and baled tyres in comparison with aggregates when percolated by leachates representative of those generated by methanogenic stage landfills. Realistic lifetime loading rates of methanogenic leachate were applied, and clogging was not generally apparent in any of the drainage media studied. This is in apparent contrast to many other studies that have demonstrated the susceptibility of all forms of drainage media to biological and chemical clogging when percolated with high strength organic and calcium rich leachates. The reasons for this difference are identified, the implications for landfill drainage system design are discussed and some suggestions for operational practice are presented for discussion. PMID:23206518

  5. Methanogenesis facilitated by geobiochemical iron cycle in a novel syntrophic methanogenic microbial community.

    PubMed

    Jiang, Shenghua; Park, Sunhwa; Yoon, Younggun; Lee, Ji-Hoon; Wu, Wei-Min; Phuoc Dan, Nguyen; Sadowsky, Michael J; Hur, Hor-Gil

    2013-09-01

    Production and emission of methane have been increasing concerns due to its significant effect on global climate change and the carbon cycle. Here we report facilitated methane production from acetate by a novel community of methanogens and acetate oxidizing bacteria in the presence of poorly crystalline akaganeite slurry. Comparative analyses showed that methanogenesis was significantly enhanced by added akaganeite and acetate was mostly stoichiometrically converted to methane. Electrons produced from anaerobic acetate oxidation are transferred to akaganeite nanorods that likely prompt the transformation into goethite nanofibers through a series of biogeochemical processes of soluble Fe(II) readsorption and Fe(III) reprecipitation. The methanogenic archaea likely harness the biotransformation of akaganeite to goethite by the Fe(III)-Fe(II) cycle to facilitate production of methane. These results provide new insights into biogeochemistry of iron minerals and methanogenesis in the environment, as well as the development of sustainable methods for microbial methane production. PMID:23919295

  6. Comparison of metabolomic profiles of microbial communities between stable and deteriorated methanogenic processes.

    PubMed

    Sasaki, Daisuke; Sasaki, Kengo; Tsuge, Yota; Morita, Masahiko; Kondo, Akihiko

    2014-11-01

    Central metabolite profiles from glucose in microbial communities during methanogenic process were compared between a stable methanogenic reactor (MR) and a deteriorated reactor (DR). The concentrations of intracellular metabolites related to the Embden-Meyerhof and pentose phosphate pathways, with the exception of pyruvate, remained high in the MR, showing increased carbon flux in the glycolysis pathway during stable methanogenesis. Extracellular acetate temporarily accumulated in the MR, consistent with higher ATP level in the MR. Intracellular concentrations of the intermediates in the reductive branch of tricarboxylic acid cycle, malate, fumarate, and succinate were higher in the DR. Low NADH/NAD(+) ratio both in the MR and DR would suggest NADH consumption during acetate and lactate/succinate production in the MR and DR, respectively. Intracellular glutamate levels were higher in the MR, correlating with lower NADPH/NADP(+) ratio concentrations in the MR. These findings contribute to a better understanding of the metabolic state during stable methanogenesis. PMID:25237777

  7. Evidence of Active Methanogen Communities in Shallow Sediments of the Sonora Margin Cold Seeps

    PubMed Central

    L'Haridon, Stéphane; Godfroy, Anne; Roussel, Erwan G.; Cragg, Barry A.; Parkes, R. John; Toffin, Laurent

    2015-01-01

    In the Sonora Margin cold seep ecosystems (Gulf of California), sediments underlying microbial mats harbor high biogenic methane concentrations, fueling various microbial communities, such as abundant lineages of anaerobic methanotrophs (ANME). However, the biodiversity, distribution, and metabolism of the microorganisms producing this methane remain poorly understood. In this study, measurements of methanogenesis using radiolabeled dimethylamine, bicarbonate, and acetate showed that biogenic methane production in these sediments was mainly dominated by methylotrophic methanogenesis, while the proportion of autotrophic methanogenesis increased with depth. Congruently, methane production and methanogenic Archaea were detected in culture enrichments amended with trimethylamine and bicarbonate. Analyses of denaturing gradient gel electrophoresis (DGGE) fingerprinting and reverse-transcribed PCR-amplified 16S rRNA sequences retrieved from these enrichments revealed the presence of active methylotrophic Methanococcoides burtonii relatives and several new autotrophic Methanogenium lineages, confirming the cooccurrence of Methanosarcinales and Methanomicrobiales methanogens with abundant ANME populations in the sediments of the Sonora Margin cold seeps. PMID:25769831

  8. Diversity and variability of methanogens during the shift from mesophilic to thermohilic conditions while biogas production.

    PubMed

    Ziembińska-Buczyńska, A; Banach, A; Bacza, T; Pieczykolan, M

    2014-12-01

    Anaerobic digestion (AD) is the most popular path of organic waste disposal. It is often used in wastewater treatment plants for excessive sludge removal. Methanogenic fermentation had usually been performed under mesophilic conditions, but in the past few years the thermophilic processes have become more popular due to economics and sludge sanitation. Methanogens, the group of microorganisms responsible for methane production, are thought to be sensitive to temperature change and it has already been proven that the communities performing methanogenesis under mesophilic and thermophilic conditions differ. But in most cases the research performed on methanogen diversity and changeability was undertaken in two separate anaerobic chambers for meso- and thermophilic conditions. It is also known that there is a group of microorganisms performing AD which are insensitive to temperature. Also the linkage between digester performance and its microbial content and community changeability is still not fully understood. That is why in this experiment we analyzed the bacterial community performing methanogenesis in a pilot scale anaerobic chamber during the shift from mesophilic to thermophilic conditions to point at the group of temperature tolerant microorganisms and their performance. The research was performed with PCR-DGGE (polymerase chain reaction-denaturing gradient gel electrophoresis). It occurred that the community biodiversity decreased together with a temperature increase. The changes were coherent for both the total bacteria community and methanogens. These bacterial shifts were also convergent with biogas production-it decreased in the beginning of the thermophilic phase with the bacterial biodiversity decrease and increased when the community seemed to be restored. DGGE results suggest that among a wide variety of microorganisms involved in AD there is a GC-rich group relatively insensitive towards temperature change, able to adapt quickly to shifts in

  9. Microbial diversity and methanogenic activity of Antrim Shale formation waters from recently fractured wells.

    PubMed

    Wuchter, Cornelia; Banning, Erin; Mincer, Tracy J; Drenzek, Nicholas J; Coolen, Marco J L

    2013-01-01

    The Antrim Shale in the Michigan Basin is one of the most productive shale gas formations in the U.S., but optimal resource recovery strategies must rely on a thorough understanding of the complex biogeochemical, microbial, and physical interdependencies in this and similar systems. We used Illumina MiSeq 16S rDNA sequencing to analyze the diversity and relative abundance of prokaryotic communities present in Antrim shale formation water of three closely spaced recently fractured gas-producing wells. In addition, the well waters were incubated with a suite of fermentative and methanogenic substrates in an effort to stimulate microbial methane generation. The three wells exhibited substantial differences in their community structure that may arise from their different drilling and fracturing histories. Bacterial sequences greatly outnumbered those of archaea and shared highest similarity to previously described cultures of mesophiles and moderate halophiles within the Firmicutes, Bacteroidetes, and δ- and ε-Proteobacteria. The majority of archaeal sequences shared highest sequence similarity to uncultured euryarchaeotal environmental clones. Some sequences closely related to cultured methylotrophic and hydrogenotrophic methanogens were also present in the initial well water. Incubation with methanol and trimethylamine stimulated methylotrophic methanogens and resulted in the largest increase in methane production in the formation waters, while fermentation triggered by the addition of yeast extract and formate indirectly stimulated hydrogenotrophic methanogens. The addition of sterile powdered shale as a complex natural substrate stimulated the rate of methane production without affecting total methane yields. Depletion of methane indicative of anaerobic methane oxidation (AMO) was observed over the course of incubation with some substrates. This process could constitute a substantial loss of methane in the shale formation. PMID:24367357

  10. Enhanced methanogenesis from hexadecane and ethylbenzene under non-methanogenic conditions

    NASA Astrophysics Data System (ADS)

    Siegert, Michael; Cichocka, Danuta; Herrmann, Steffi; Richnow, Hans-Hermann; Springael, Dirk; Krüger, Martin

    2010-05-01

    Microbially enhanced oil recovery (MEOR) may provide access to remaining, but yet inaccessible petroleum in reservoirs. The microbial conversion of heavy hydrocarbon remnants into gaseous methane could at least provide access to energy which would otherwise be lost. On the other hand, methanogenesis could remove toxic hydrocarbons from contaminated aquifers and sediments. Therefore, sediment samples from a contaminated sea port basin were investigated to assess the in situ potential for methanogenic hydrocarbon degradation. Since this process is believed to be a sequential syntrophic procedure, non-methanogenic conditions were created in sediment microcosms to facilitate the first hydrocarbon attacking step. To achieve this, a high electron potential was created by the addition of ferrihydrite, manganese oxide, nitrate or sulfate as electron acceptors. Hexadecane, ethylbenzene or naphthalene were used as model carbon substrates. Methanogenesis evolved rapidly from set ups treated with iron and manganese, but not nitrate, reflecting the in situ conditions at the site. Surprisingly, on sulfate methanogenesis was neither inhibited nor significantly supported. Methane formation rates were the highest with hexadecane as substrate, followed by ethylbenzene and naphthalene. Methane was removed in high rates at the same time by anaerobic methanotrophs. The microbial community in situ and in vitro was dominated by members of the Geobacteraceae. Their methanogenic partners were quantified, targeting the genes encoding for the methyl coenzyme M reductase (mcrA). Methane consumption in the microcosms and the presence of methanotrophic anaerobes belonging to the ANME-1 and ANME-2 clusters suggest anaerobic methanotrophy as an accompanying process. mcrA genes belonging to the ANME-1 & -2 clusters were detected in lower copy numbers than the methanogenic mcrA, which is in good agreement with the activity measurements. These results indicate that the in situ stimulation of

  11. Methanogenesis in a thermophilic (58 degrees C) anaerobic digestor: Methanothrix sp. as an important aceticlastic methanogen

    SciTech Connect

    Zinder, S.H.; Cardwell, S.C.; Anguish, T.; Lee, M.; Koch, M.

    1984-04-01

    Aceticlastic methanogens and other microbial groups were enumerated in a 58 degrees C laboratory-scale (3 liter) anaerobic digestor which was fed air-classified municipal refuse, a lignocellulosic waste (loading rate = 1.8 to 2.7 g of volatile solids per liter per day: retention time = 10 days). Two weeks after start-up, Methanosarcina sp. was present in high numbers (10/sup 5/ to 10/sup 6/ CFU/ml) and autofluorescent Methanosarcina clumps were abundant in sludge. After 4 months of operation, numbers of Methanosarcina sp. dropped 2 to 3 orders of magnitude and large numbers (most probable number = 10/sup 6/ to 10/sup 7/ /ml) Methanothrix sp. were found. Methanothrix sp. had apparently displaced Methanosarcina sp. as the dominant aceticlastic methanogen. During the period when Methanothrix sp. was dominant, acetate concentrations varied between 0.3 and 1.5 mumol/ml during the daily feeding cycle, and acetate was the precursor of 63 to 66% of the methane produced during peak digestor methanogenesis. The apparent Km value obtained for methanogenesis from acetate, 0.3 mumol/ml, indicated that the aceticlastic methanogens were saturated for substrate during most of the digestor cycle. CO2 reducing methanogens were capable of methanogenesis at rates more than 12 times greater than those usually found in the digestor. Added propionate (4.5 mumol/ml) was metabolized slowly and slightly inhibited methanogenesis. Added n-butyrate, isobutyrate, or n-valerate (4.5 mumol/ml each) were broken down within 24 h. Isobutyrate was oxidized to acetate, a novel reaction possibly involving isomerization to n-butyrate. The rapid growth rate and versatile metabolism of Methanosarcina sp. make it a likely organism to be involved in start-up whereas the low Km value of Methanothrix sp. for acetate may cause it to be favored in stable digestors operated with long retention times.

  12. Propidium monoazide treatment to distinguish between live and dead methanogens in pure cultures and environmental samples.

    PubMed

    Heise, Janine; Nega, Marcella; Alawi, Mashal; Wagner, Dirk

    2016-02-01

    In clinical trials investigating human health and in the analysis of microbial communities in cultures and natural environments, it is a substantial challenge to differentiate between living, potentially active communities and dead cells. The DNA-intercalating dye propidium monoazide (PMA) enables the selective masking of DNA from dead, membrane-compromised cells immediately before DNA extraction. In the present study, we evaluated for the first time a PMA treatment for methanogenic archaea in cultures and particle-rich environmental samples. Using microscopic analyses, we confirmed the applicability of the LIVE/DEAD(®) BacLight™ kit to methanogenic archaea and demonstrated the maintenance of intact cell membranes of methanogens in the presence of PMA. Although strain-specific differences in the efficiency of PMA treatment to methanogenic archaea were observed, we developed an optimal procedure using 130 μM PMA and 5min of photo-activation with blue LED light. The results showed that the effectiveness of the PMA treatment strongly depends on the texture of the sediment/soil: silt and clay-rich sediments represent a challenge at all concentrations, whereas successful suppression of DNA from dead cells with compromised membranes was possible for low particle loads of sandy soil (total suspended solids (TSS)≤200 mg mL(-1)). Conclusively, we present two strategies to overcome the problem of insufficient light activation of PMA caused by the turbidity effect (shielding) in particle-rich environmental samples by (i) dilution of the particle-rich sample and (ii) detachment of the cells and the free DNA from the sediment prior to a PMA treatment. Both strategies promise to be usable options for distinguishing living cells and free DNA in complex environmental samples. PMID:26656002

  13. Microbial diversity and methanogenic activity of Antrim Shale formation waters from recently fractured wells

    PubMed Central

    Wuchter, Cornelia; Banning, Erin; Mincer, Tracy J.; Drenzek, Nicholas J.; Coolen, Marco J. L.

    2013-01-01

    The Antrim Shale in the Michigan Basin is one of the most productive shale gas formations in the U.S., but optimal resource recovery strategies must rely on a thorough understanding of the complex biogeochemical, microbial, and physical interdependencies in this and similar systems. We used Illumina MiSeq 16S rDNA sequencing to analyze the diversity and relative abundance of prokaryotic communities present in Antrim shale formation water of three closely spaced recently fractured gas-producing wells. In addition, the well waters were incubated with a suite of fermentative and methanogenic substrates in an effort to stimulate microbial methane generation. The three wells exhibited substantial differences in their community structure that may arise from their different drilling and fracturing histories. Bacterial sequences greatly outnumbered those of archaea and shared highest similarity to previously described cultures of mesophiles and moderate halophiles within the Firmicutes, Bacteroidetes, and δ- and ε-Proteobacteria. The majority of archaeal sequences shared highest sequence similarity to uncultured euryarchaeotal environmental clones. Some sequences closely related to cultured methylotrophic and hydrogenotrophic methanogens were also present in the initial well water. Incubation with methanol and trimethylamine stimulated methylotrophic methanogens and resulted in the largest increase in methane production in the formation waters, while fermentation triggered by the addition of yeast extract and formate indirectly stimulated hydrogenotrophic methanogens. The addition of sterile powdered shale as a complex natural substrate stimulated the rate of methane production without affecting total methane yields. Depletion of methane indicative of anaerobic methane oxidation (AMO) was observed over the course of incubation with some substrates. This process could constitute a substantial loss of methane in the shale formation. PMID:24367357

  14. Limited aeration of methanogenic systems for treatment of sulfate-containing wastewater

    SciTech Connect

    Zitomer, D.H.; Shrout, J.D.

    1998-07-01

    Wastewaters with high sulfate concentrations are often not readily amenable to methanogenic treatment due to production of potentially inhibitory hydrogen sulfide. Some reports indicate that treatment has been enhanced by air sparging of recycle flows to air-strip hydrogen sulfide and create a selective environment for reduction of sulfate to elemental sulfur, rather than sulfides and thiosulfate. In this report, direct aeration of methanogenic expanded beds was employed for treatment of high-sulfate wastewater and compared to a strictly anaerobic control. An influent wastewater COD:SO{sup {minus}2}{sub 4} ratio as low as 4,4:1 did not discernibly inhibit methane production in the anaerobic control. At a relatively high OLR of 30g COD/L{sub A}-d and a relatively short HRT of 4.2 hours, the COD removal of the control was 97%. Methanogenesis was also sustained in the aerated reactors which had oxygen transfer rates of between 2 to 5% of the OLR. These aerated reactors converted approximately 70% of the COD to methane. Future research will involve higher sulfate loadings and aeration rates to determine possible benefits of direct aeration of methanogenic expanded beds.

  15. Choline and N,N-Dimethylethanolamine as Direct Substrates for Methanogens

    PubMed Central

    Watkins, Andrew J.; Roussel, Erwan G.; Webster, Gordon; Parkes, R. John

    2012-01-01

    Choline (N,N,N-trimethylethanolamine), which is widely distributed in membrane lipids and is a component of sediment biota, has been shown to be utilized anaerobically by mixed prokaryote cultures to produce methane but not by pure cultures of methanogens. Here, we show that five recently isolated Methanococcoides strains from a range of sediments (Aarhus Bay, Denmark; Severn Estuary mudflats at Portishead, United Kingdom; Darwin Mud Volcano, Gulf of Cadiz; Napoli mud volcano, eastern Mediterranean) can directly utilize choline for methanogenesis producing ethanolamine, which is not further metabolized. Di- and monomethylethanolamine are metabolic intermediates that temporarily accumulate. Consistent with this, dimethylethanolamine was shown to be another new growth substrate, but monomethylethanolamine was not. The specific methanogen inhibitor 2-bromoethanesulfonate (BES) inhibited methane production from choline. When choline and trimethylamine are provided together, diauxic growth occurs, with trimethylamine being utilized first, and then after a lag (∼7 days) choline is metabolized. Three type strains of Methanococcoides (M. methylutens, M. burtonii, and M. alaskense), in contrast, did not utilize choline. However, two of them (M. methylutens and M. burtonii) did metabolize dimethylethanolamine. These results extend the known substrates that can be directly utilized by some methanogens, giving them the advantage that they would not be reliant on bacterial syntrophs for their substrate supply. PMID:23001649

  16. In vitro Evaluation of Different Feeds for Their Potential to Generate Methane and Change Methanogen Diversity

    PubMed Central

    Kim, Seon-Ho; Mamuad, Lovelia L.; Jeong, Chang-Dae; Choi, Yeon-Jae; Lee, Sung Sill; Ko, Jong-Youl; Lee, Sang-Suk

    2013-01-01

    Optimization of the dietary formulation is the most effective way to reduce methane. Nineteen feed ingredients (brans, vegetable proteins, and grains) were evaluated for their potential to generate methane and change methanogen diversity using an in vitro ruminal fermentation technique. Feed formulations categorized into high, medium and low production based on methane production of each ingredient were then subjected to in vitro fermentation to determine the real methane production and their effects on digestibility. Methanogen diversity among low, medium and high-methane producing groups was analyzed by PCR-DGGE. The highest methane production was observed in Korean wheat bran, soybean and perilla meals, and wheat and maize of brans, vegetable protein and cereal groups, respectively. On the other hand, corn bran, cotton seed meal and barley led to the lowest production in the same groups. Nine bacteria and 18 methanogen 16s rDNA PCR-DGGE dominant bands were identified with 83% to 99% and 92% to 100% similarity, respectively. Overall, the results of this study showed that methane emissions from ruminants can be mitigated through proper selection of feed ingredients to be used in the formulation of diets. PMID:25049760

  17. Anaerobic metabolism of nitroaromatic compounds by sulfate-reducing and methanogenic bacteria

    SciTech Connect

    Boopathy, R.; Kulpa, C.F.

    1994-06-01

    Ecological observations suggest that sulfate-reducing and methanogenic bacteria might metabolize nitroaromatic compounds under anaerobic conditions if appropriate electron donors and electron acceptors are present in the environment, but this ability had not been demonstrated until recently. Most studies on the microbial metabolism of nitroaromatic compounds used aerobic microorganisms. In most cases no mineralization of nitroaromatics occurs, and only superficial modifications of the structures are reported. However, under anaerobic sulfate-reducing conditions, the nitroaromatic compounds reportedly undergo a series of reductions with the formation of amino compounds. For example, trinitrotoluene under sulfate-reducing conditions is reduced to triaminotoluene by the enzyme nitrite reductase, which is commonly found in many Desulfovibrio spp. The removal of ammonia from triaminotoluene is achieved by reductive deamination catalyzed by the enzyme reductive deaminase, with the production of ammonia and toluene. Some sulfate reducers can metabolize toluene to CO{sub 2}. Similar metabolic processes could be applied to other nitroaromatic compounds like nitrobenzene, nitrobenzoic acids, nitrophenols, and aniline. Many methanogenic bacteria can reduce nitroaromatic compounds to amino compounds. In this paper we review the anaerobic metabolic processes of nitroaromatic compounds under sulfate-reducing And methanogenic conditions.

  18. Biodegradation of C7 and C8 iso-alkanes under methanogenic conditions.

    PubMed

    Abu Laban, Nidal; Dao, Anh; Semple, Kathleen; Foght, Julia

    2015-12-01

    Iso-alkanes comprise a substantial proportion of petroleum and refined products that impact the environment, but their fate is cryptic under methanogenic conditions. We investigated methanogenic biodegradation of C7 and C8 iso-alkanes found in naphtha, specifically 2-methylhexane, 3-methylhexane, 2-methylheptane, 4-methylheptane and 3-ethylhexane. These were incubated as a mixture or individually with enrichment cultures derived from oil sands tailings ponds that generate methane from naphtha components; substrate depletion and methane production were monitored for up to 663 days. 3-Methylhexane and 4-methylheptane were degraded both singly and in the mixture, whereas 2-methylhexane and 2-methylheptane resisted degradation as single substrates but were depleted in the iso-alkane mixture, suggesting co-metabolism. 3-Ethylhexane was degraded neither singly nor with co-substrates. Putative metabolites consistent with succinylated C7 and C8 were detected, suggesting activation by addition of iso-alkanes to fumarate and corresponding to detection of alkylsuccinate synthase-like genes. 454 pyrotag sequencing, cloning and terminal restriction fragment length polymorphism of 16S rRNA genes revealed predominance of a novel member of the family Peptococcaceae (order Clostridiales) and Archaea affiliated with Methanoregula and Methanosaeta. We report here isomer-specific metabolism of C7 -C8 iso-alkanes under methanogenic conditions and propose their activation by a novel Peptococcaceae via addition to fumarate. PMID:25331365

  19. Conductive iron oxide minerals accelerate syntrophic cooperation in methanogenic benzoate degradation.

    PubMed

    Zhuang, Li; Tang, Jia; Wang, Yueqiang; Hu, Min; Zhou, Shungui

    2015-08-15

    Recent studies have suggested that conductive iron oxide minerals can facilitate syntrophic metabolism of the methanogenic degradation of organic matter, such as ethanol, propionate and butyrate, in natural and engineered microbial ecosystems. This enhanced syntrophy involves direct interspecies electron transfer (DIET) powered by microorganisms exchanging metabolic electrons through electrically conductive minerals. Here, we evaluated the possibility that conductive iron oxides (hematite and magnetite) can stimulate the methanogenic degradation of benzoate, which is a common intermediate in the anaerobic metabolism of aromatic compounds. The results showed that 89-94% of the electrons released from benzoate oxidation were recovered in CH4 production, and acetate was identified as the only carbon-bearing intermediate during benzoate degradation. Compared with the iron-free controls, the rates of methanogenic benzoate degradation were enhanced by 25% and 53% in the presence of hematite and magnetite, respectively. This stimulatory effect probably resulted from DIET-mediated methanogenesis in which electrons transfer between syntrophic partners via conductive iron minerals. Phylogenetic analyses revealed that Bacillaceae, Peptococcaceae, and Methanobacterium are potentially involved in the functioning of syntrophic DIET. Considering the ubiquitous presence of iron minerals within soils and sediments, the findings of this study will increase the current understanding of the natural biological attenuation of aromatic hydrocarbons in anaerobic environments. PMID:25827267

  20. Methane Production on Rock and Soil Substrates by Methanogens: Implications for Life on Mars

    NASA Astrophysics Data System (ADS)

    Kozup, H. A.; Kral, T. A.

    2009-12-01

    In order to understand the methanogens as models for possible life on Mars, and some of the factors likely to be important in determining their abundance and distribution, we have measured their ability to produce methane on a few types of inorganic rock and soil substrates. Since organic materials have not been detected in measurable quantities at the surface of Mars, there is no reason to believe that they would exist in the subsurface. Samples of three methanogens (Methanosarcina barkeri, Methanobacterium formicicum, and Methanothermobacter wolfeii) were placed on four substrates (sand, gravel, basalt, and a Mars soil simulant, JSC Mars-1) and methane production measured. Glass beads were used as a control substrate. As in earlier experiments with JSC Mars-1 soil simulant, a crushed volcanic tephra, methane was produced by all three methanogens when placed on the substrates, sand and gravel. None produced methane on basalt in these experiments, a mineral common in Martian soil. While these substrates do not represent the full range of materials likely to be present on the surface of Mars, the present results suggest that while some surface materials on Mars may not support this type of organism, others might.

  1. Methanogen Diversity in Indigenous and Introduced Ruminant Species on the Tibetan Plateau.

    PubMed

    Huang, Xiao Dan; Martinez-Fernandez, Gonzalo; Padmanabha, Jagadish; Long, Ruijun; Denman, Stuart E; McSweeney, Christopher S

    2016-01-01

    Host factors are regarded as important in shaping the archaeal community in the rumen but few controlled studies have been performed to demonstrate this across host species under the same environmental conditions. A study was designed to investigate the structure of the methanogen community in the rumen of two indigenous (yak and Tibetan sheep) and two introduced domestic ruminant (cattle and crossbred sheep) species raised and fed under similar conditions on the high altitude Tibetan Plateau. The methylotrophic Methanomassiliicoccaceae was the predominant archaeal group in all animals even though Methanobrevibacter are usually present in greater abundance in ruminants globally. Furthermore, within the Methanomassiliicoccaceae family members from Mmc. group 10 and Mmc. group 4 were dominant in Tibetan Plateau ruminants compared to Mmc. group 12 found to be highest in other ruminants studied. Small ruminants presented the highest number of sequences that belonged to Methanomassiliicoccaceae compared to the larger ruminants. Although the methanogen community structure was different among the ruminant species, there were striking similarities between the animals in this environment. This indicates that factors such as the extreme environmental conditions and diet on the Tibetan Plateau might have a greater impact on rumen methanogen community compared to host differences. PMID:27274707

  2. Anaerobic digestion of renewable biomass: thermophilic temperature governs methanogen population dynamics.

    PubMed

    Krakat, Niclas; Westphal, A; Schmidt, S; Scherer, P

    2010-03-01

    Beet silage and beet juice were digested continuously as representative energy crops in a thermophilic biogas fermentor for more than 7 years. Fluorescence microscopy of 15 samples covering a period of 650 days revealed that a decrease in temperature from 60 degrees C to 55 degrees C converted a morphologically uniform archaeal population (rods) into a population of methanogens exhibiting different cellular morphologies (rods and coccoid cells). A subsequent temperature increase back to 60 degrees C reestablished the uniform morphology of methanogens observed in the previous 60 degrees C period. In order to verify these observations, representative samples were investigated by amplified rRNA gene restriction analysis (ARDRA) and fluorescence in situ hybridization (FISH). Both methods confirmed the temperature-dependent population shift observed by fluorescence microscopy. Moreover, all samples investigated demonstrated that hydrogenotrophic Methanobacteriales dominated in the fermentor, as 29 of 34 identified operational taxonomic units (OTUs) were assigned to this order. This apparent discrimination of acetoclastic methanogens contradicts common models for anaerobic digestion processes, such as anaerobic digestion model 1 (ADM1), which describes the acetotrophic Euryarchaeota as predominant organisms. PMID:20097828

  3. RDX biodegradation by a methanogenic enrichment culture obtained from an explosives manufacturing wastewater treatment plant

    SciTech Connect

    Adrian, N.R.; Sutherland, K.

    1998-12-01

    This study examined the biodegradation of RDX in wastewater from an industrial wastewater treatment plant at the Holston Army Ammunition Plant in Kingsport, TN. Serum bottles containing 100 ml of a basal salts medium amended with 10 percent (v/v) sludge from the anoxic filter at the plant were amended with RDX and incubated under methanogenic conditions. Biodegradation intermediates corresponding to the mono-, di-, and trinitroso- RDX were observed. A methanogenic enrichment culture, derived from the wastewater, biodegraded 25% micrometer RDX in less than 16 days when ethanol was supplied as an electron donor. Methane production in the ethanol amended bottles was only observed after RDX had been depleted, while RDX unamended controls experienced no lag in methane production. The addition of 5 mM BESA to the culture inhibited methane production, but not RDX and ethanol degradation. These findings demonstrate the importance of adding reduced cosubstrates to enhance RDX biodegradation, and support the hypothesis that RDX is serving as a terminal electron acceptor in methanogenic environments.

  4. Significance of ether lipids derived from methanogens in sediments from modern and ancient basins

    SciTech Connect

    Shouten, S.; Sinninghe Damste, J.S.; Hoefs, M.J.L.; De Leeuw, J.W. Netherlands and Utrecht Univ. )

    1996-01-01

    High amounts of biphytanyl carbon skeletons with 0-3 cyclopentyl rings and low amounts of 2, 6, 10, 14, 18-pentamethyleicosane (PME) were detected after treatment with hydrogen iodide of fractions of several sediment extracts. These compounds are biosynthesized by archaebacteria only. Since the sediments were deposited in normal marine environments, these lipids are derived from methanogenic bacteria and more specifically Methanosarcina barkeri. In recent sediments from in and below the extensive oxygen-minimum zone in the Arabian Sea high amounts were detected in surface sediment samples where oxygen is still present. Our data suggest that these ether-bound compounds may be indicators for methane-production in the upper part of the water column and possibly an extensive oxygen-minimum zone. The concentrations varied strongly (2 orders of magnitude) in the Miocene Monterey Formation suggesting significant variations in water column methanogenic activity. The stable carbon isotopic compositions of the acyclic and cyclic biphytane carbon skeletons are several per mil heavier than those of compounds derived from photoautotrophic organisms whereas the stable carbon isotopic compositions of PME are several per mil lighter. This indicates the presence of different species of methanogens utilizing different carbon substrates.

  5. Significance of ether lipids derived from methanogens in sediments from modern and ancient basins

    SciTech Connect

    Shouten, S.; Sinninghe Damste, J.S.; Hoefs, M.J.L.; De Leeuw, J.W. |

    1996-12-31

    High amounts of biphytanyl carbon skeletons with 0-3 cyclopentyl rings and low amounts of 2, 6, 10, 14, 18-pentamethyleicosane (PME) were detected after treatment with hydrogen iodide of fractions of several sediment extracts. These compounds are biosynthesized by archaebacteria only. Since the sediments were deposited in normal marine environments, these lipids are derived from methanogenic bacteria and more specifically Methanosarcina barkeri. In recent sediments from in and below the extensive oxygen-minimum zone in the Arabian Sea high amounts were detected in surface sediment samples where oxygen is still present. Our data suggest that these ether-bound compounds may be indicators for methane-production in the upper part of the water column and possibly an extensive oxygen-minimum zone. The concentrations varied strongly (2 orders of magnitude) in the Miocene Monterey Formation suggesting significant variations in water column methanogenic activity. The stable carbon isotopic compositions of the acyclic and cyclic biphytane carbon skeletons are several per mil heavier than those of compounds derived from photoautotrophic organisms whereas the stable carbon isotopic compositions of PME are several per mil lighter. This indicates the presence of different species of methanogens utilizing different carbon substrates.

  6. Magnetite particles triggering a faster and more robust syntrophic pathway of methanogenic propionate degradation.

    PubMed

    Cruz Viggi, Carolina; Rossetti, Simona; Fazi, Stefano; Paiano, Paola; Majone, Mauro; Aulenta, Federico

    2014-07-01

    Interspecies electron transfer mechanisms between Bacteria and Archaea play a pivotal role during methanogenic degradation of organic matter in natural and engineered anaerobic ecosystems. Growing evidence suggests that in syntrophic communities electron transfer does not rely exclusively on the exchange of diffusible molecules and energy carriers such as hydrogen or formate, rather microorganisms have the capability to exchange metabolic electrons in a more direct manner. Here, we show that supplementation of micrometer-size magnetite (Fe3O4) particles to a methanogenic sludge enhanced (up to 33%) the methane production rate from propionate, a key intermediate in the anaerobic digestion of organic matter and a model substrate to study energy-limited syntrophic communities. The stimulatory effect most probably resulted from the establishment of a direct interspecies electron transfer (DIET), based on magnetite particles serving as electron conduits between propionate-oxidizing acetogens and carbon dioxide-reducing methanogens. Theoretical calculations revealed that DIET allows electrons to be transferred among syntrophic partners at rates which are substantially higher than those attainable via interspecies H2 transfer. Besides the remarkable potential for improving anaerobic digestion, which is a proven biological strategy for renewable energy production, the herein described conduction-based DIET could also have a role in natural methane emissions from magnetite-rich soils and sediments. PMID:24901501

  7. Choline and N,N-dimethylethanolamine as direct substrates for methanogens.

    PubMed

    Watkins, Andrew J; Roussel, Erwan G; Webster, Gordon; Parkes, R John; Sass, Henrik

    2012-12-01

    Choline (N,N,N-trimethylethanolamine), which is widely distributed in membrane lipids and is a component of sediment biota, has been shown to be utilized anaerobically by mixed prokaryote cultures to produce methane but not by pure cultures of methanogens. Here, we show that five recently isolated Methanococcoides strains from a range of sediments (Aarhus Bay, Denmark; Severn Estuary mudflats at Portishead, United Kingdom; Darwin Mud Volcano, Gulf of Cadiz; Napoli mud volcano, eastern Mediterranean) can directly utilize choline for methanogenesis producing ethanolamine, which is not further metabolized. Di- and monomethylethanolamine are metabolic intermediates that temporarily accumulate. Consistent with this, dimethylethanolamine was shown to be another new growth substrate, but monomethylethanolamine was not. The specific methanogen inhibitor 2-bromoethanesulfonate (BES) inhibited methane production from choline. When choline and trimethylamine are provided together, diauxic growth occurs, with trimethylamine being utilized first, and then after a lag (∼7 days) choline is metabolized. Three type strains of Methanococcoides (M. methylutens, M. burtonii, and M. alaskense), in contrast, did not utilize choline. However, two of them (M. methylutens and M. burtonii) did metabolize dimethylethanolamine. These results extend the known substrates that can be directly utilized by some methanogens, giving them the advantage that they would not be reliant on bacterial syntrophs for their substrate supply. PMID:23001649

  8. Methanogen Diversity in Indigenous and Introduced Ruminant Species on the Tibetan Plateau

    PubMed Central

    Huang, Xiao Dan; Martinez-Fernandez, Gonzalo; Padmanabha, Jagadish; Long, Ruijun; Denman, Stuart E.; McSweeney, Christopher S.

    2016-01-01

    Host factors are regarded as important in shaping the archaeal community in the rumen but few controlled studies have been performed to demonstrate this across host species under the same environmental conditions. A study was designed to investigate the structure of the methanogen community in the rumen of two indigenous (yak and Tibetan sheep) and two introduced domestic ruminant (cattle and crossbred sheep) species raised and fed under similar conditions on the high altitude Tibetan Plateau. The methylotrophic Methanomassiliicoccaceae was the predominant archaeal group in all animals even though Methanobrevibacter are usually present in greater abundance in ruminants globally. Furthermore, within the Methanomassiliicoccaceae family members from Mmc. group 10 and Mmc. group 4 were dominant in Tibetan Plateau ruminants compared to Mmc. group 12 found to be highest in other ruminants studied. Small ruminants presented the highest number of sequences that belonged to Methanomassiliicoccaceae compared to the larger ruminants. Although the methanogen community structure was different among the ruminant species, there were striking similarities between the animals in this environment. This indicates that factors such as the extreme environmental conditions and diet on the Tibetan Plateau might have a greater impact on rumen methanogen community compared to host differences. PMID:27274707

  9. [Artificial cultivation modes for Dendrobium officinale].

    PubMed

    Si, Jin-Ping; Yu, Qiao-Xian; Song, Xian-Shui; Shao, Wei-Jiang

    2013-02-01

    Since the beginning of the new century, the artificial cultivation of Dendrobium officinale has made a breakthrough progress. This paper systematically expounds key technologies, main features and cautions of the cultivation modes e.g. bionic-facility cultivation, the original ecological cultivation, and potting cultivation for D. officinale, which can provide useful information for the development and improvement of D. officinale industry. PMID:23713268

  10. Time-Resolved DNA Stable Isotope Probing Links Desulfobacterales- and Coriobacteriaceae-Related Bacteria to Anaerobic Degradation of Benzene under Methanogenic Conditions

    PubMed Central

    Noguchi, Mana; Kurisu, Futoshi; Kasuga, Ikuro; Furumai, Hiroaki

    2014-01-01

    To identify the microorganisms involved in benzene degradation, DNA-stable isotope probing (SIP) with 13C-benzene was applied to a methanogenic benzene-degrading enrichment culture. Pyrosequencing of ribosomal RNA (rRNA) gene sequences revealed that the community structure was highly complex in spite of a 3-year incubation only with benzene. The culture degraded 98% of approximately 1 mM 13C-benzene and mineralized 72% of that within 63 d. The terminal restriction fragment length polymorphism (T-RFLP) profiles of the buoyant density fractions revealed the incorporation of 13C into two phylotypes after 64 d. These two phylotypes were determined to be Desulfobacterales- and Coriobacteriaceae-related bacteria by cloning and sequencing of the 16S rRNA gene in the 13C-labeled DNA abundant fraction. Comparative pyrosequencing analysis of the buoyant density fractions of 12C- and 13C-labeled samples indicated the incorporation of 13C into three bacterial and one archaeal OTUs related to Desulfobacterales, Coriobacteriales, Rhodocyclaceae, and Methanosarcinales. The first two OTUs included the bacteria detected by T-RFLP-cloning-sequencing analysis. Furthermore, time-resolved SIP analysis confirmed that the activity of all these microbes appeared at the earliest stage of degradation. In this methanogenic culture, Desulfobacterales- and Coriobacteriaceae-related bacteria were most likely to be the major benzene degraders. PMID:24909708

  11. DNA stable-isotope probing of oil sands tailings pond enrichment cultures reveals different key players for toluene degradation under methanogenic and sulfidogenic conditions.

    PubMed

    Laban, Nidal Abu; Dao, Anh; Foght, Julia

    2015-05-01

    Oil sands tailings ponds are anaerobic repositories of fluid wastes produced by extraction of bitumen from oil sands ores. Diverse indigenous microbiota biodegrade hydrocarbons (including toluene) in situ, producing methane, carbon dioxide and/or hydrogen sulfide, depending on electron acceptor availability. Stable-isotope probing of cultures enriched from tailings associated specific taxa and functional genes to (13)C6- and (12)C7-toluene degradation under methanogenic and sulfate-reducing conditions. Total DNA was subjected to isopycnic ultracentrifugation followed by gradient fraction analysis using terminal restriction fragment length polymorphism (T-RFLP) and construction of 16S rRNA, benzylsuccinate synthase (bssA) and dissimilatory sulfite reductase (dsrB) gene clone libraries. T-RFLP analysis plus sequencing and in silico digestion of cloned taxonomic and functional genes revealed that Clostridiales, particularly Desulfosporosinus (136 bp T-RF) contained bssA genes and were key toluene degraders during methanogenesis dominated by Methanosaeta. Deltaproteobacterial Desulfobulbaceae (157 bp T-RF) became dominant under sulfidogenic conditions, likely because the Desulfosporosinus T-RF 136 apparently lacks dsrB and therefore, unlike its close relatives, is presumed incapable of dissimilatory sulfate reduction. We infer incomplete oxidation of toluene by Desulfosporosinus in syntrophic association with Methanosaeta under methanogenic conditions, and complete toluene oxidation by Desulfobulbaceae during sulfate reduction. PMID:25873466

  12. Slash and Burn Agriculture: A Dynamic Spatio-temporal Model of Shifting Cultivation Locations and Areas

    NASA Astrophysics Data System (ADS)

    Plagge, C. E.; Frolking, S.; Chini, L. P.; Hurtt, G.

    2008-12-01

    Shifting cultivation is a form of agriculture, also known as slash-and-burn or swidden agriculture, in which a plot of forest is cleared and then cultivated continuously for several years, after which it is abandoned to revert to natural vegetation, and then is subsequently re-cleared after a longer fallow period. Shifting cultivation is an important form of agriculture because it affects soil erosion rates, canopy cover in tropical forests, nutrient deficiency in soils, and also has an impact on the global carbon cycle. Because it is generally outside of the larger economy, shifting cultivation is not well-represented in large-scale earth system analyses. We investigated a new way to model shifting cultivation which will be included in a global land-use transitions model to better quantify this type of land use, both historically and into the future. Ultimately this study will improve simulations of changes in the Earth system and will aid in the study of the carbon cycle and thus climate change. Our model calculates the area of shifting cultivation in square kilometers per half-degree grid cell, using gridded population data, the fraction of that population that is rural, the fraction of global population that practices shifting cultivation, the crop area needed per person, and the length of cultivation plus the fallow. Locations of shifting cultivation were further constrained by variables such as potential vegetation biomass density, population density, fraction of land already in use, GDP per capita, and average winter temperatures. With this model, we generated global estimates for total cultivated area, total population involved in shifting cultivation, and total shifting cultivation area including fallow lands. From this model it was estimated that the total global area of shifting cultivation in 2000 was approximately 1.5 million km2 with 90,000 km2 of that actually in cultivation by 190 million people.

  13. Microbiological characterization and specific methanogenic activity of anaerobe sludges used in urban solid waste treatment

    SciTech Connect

    Sandoval Lozano, Claudia Johanna Vergara Mendoza, Marisol; Carreno de Arango, Mariela; Castillo Monroy, Edgar Fernando

    2009-02-15

    This study presents the microbiological characterization of the anaerobic sludge used in a two-stage anaerobic reactor for the treatment of organic fraction of urban solid waste (OFUSW). This treatment is one alternative for reducing solid waste in landfills at the same time producing a biogas (CH{sub 4} and CO{sub 2}) and an effluent that can be used as biofertilizer. The system was inoculated with sludge from a wastewater treatment plant (WWTP) (Rio Frio Plant in Bucaramanga-Colombia) and a methanogenic anaerobic digester for the treatment of pig manure (Mesa de los Santos in Santander). Bacterial populations were evaluated by counting groups related to oxygen sensitivity, while metabolic groups were determined by most probable number (MPN) technique. Specific methanogenic activity (SMA) for acetate, formate, methanol and ethanol substrates was also determined. In the acidogenic reactor (R1), volatile fatty acids (VFA) reached values of 25,000 mg L{sup -1} and a concentration of CO{sub 2} of 90%. In this reactor, the fermentative population was predominant (10{sup 5}-10{sup 6} MPN mL{sup -1}). The acetogenic population was (10{sup 5} MPN mL{sup -1}) and the sulphate-reducing population was (10{sup 4}-10{sup 5} MPN mL{sup -1}). In the methanogenic reactor (R2), levels of CH{sub 4} (70%) were higher than CO{sub 2} (25%), whereas the VFA values were lower than 4000 mg L{sup -1}. Substrate competition between sulphate-reducing (10{sup 4}-10{sup 5} MPN mL{sup -1}) and methanogenic bacteria (10{sup 5} MPN mL{sup -1}) was not detected. From the SMA results obtained, acetoclastic (2.39 g COD-CH{sub 4} g{sup -1} VSS{sup -1} day{sup -1}) and hydrogenophilic (0.94 g COD-CH{sub 4} g{sup -1} VSS{sup -1} day{sup -1}) transformations as possible metabolic pathways used by methanogenic bacteria is suggested from the SMA results obtained. Methanotrix sp., Methanosarcina sp., Methanoccocus sp. and Methanobacterium sp. were identified.

  14. Microbiological characterization and specific methanogenic activity of anaerobe sludges used in urban solid waste treatment.

    PubMed

    Lozano, Claudia Johanna Sandoval; Mendoza, Marisol Vergara; de Arango, Mariela Carreño; Monroy, Edgar Fernando Castillo

    2009-02-01

    This study presents the microbiological characterization of the anaerobic sludge used in a two-stage anaerobic reactor for the treatment of organic fraction of urban solid waste (OFUSW). This treatment is one alternative for reducing solid waste in landfills at the same time producing a biogas (CH(4) and CO(2)) and an effluent that can be used as biofertilizer. The system was inoculated with sludge from a wastewater treatment plant (WWTP) (Río Frío Plant in Bucaramanga-Colombia) and a methanogenic anaerobic digester for the treatment of pig manure (Mesa de los Santos in Santander). Bacterial populations were evaluated by counting groups related to oxygen sensitivity, while metabolic groups were determined by most probable number (MPN) technique. Specific methanogenic activity (SMA) for acetate, formate, methanol and ethanol substrates was also determined. In the acidogenic reactor (R1), volatile fatty acids (VFA) reached values of 25,000 mg L(-1) and a concentration of CO(2) of 90%. In this reactor, the fermentative population was predominant (10(5)-10(6)MPN mL(-1)). The acetogenic population was (10(5)MPN mL(-1)) and the sulphate-reducing population was (10(4)-10(5)MPN mL(-1)). In the methanogenic reactor (R2), levels of CH(4) (70%) were higher than CO(2) (25%), whereas the VFA values were lower than 4000 mg L(-1). Substrate competition between sulphate-reducing (10(4)-10(5)MPN mL(-1)) and methanogenic bacteria (10(5)MPN mL(-1)) was not detected. From the SMA results obtained, acetoclastic (2.39 g COD-CH(4)g(-1)VSS(-1)day(-1)) and hydrogenophilic (0.94 g COD-CH(4)g(-1)VSS(-1)day(-1)) transformations as possible metabolic pathways used by methanogenic bacteria is suggested from the SMA results obtained. Methanotrix sp., Methanosarcina sp., Methanoccocus sp. and Methanobacterium sp. were identified. PMID:18707861

  15. Genotypic diversity among rhizospheric bacteria of three legumes assessed by cultivation-dependent and cultivation-independent techniques.

    PubMed

    Pongsilp, Neelawan; Nimnoi, Pongrawee; Lumyong, Saisamorn

    2012-02-01

    The genotypic diversity of rhizospheric bacteria of 3 legumes including Vigna radiata, Arachis hypogaea and Acacia mangium was compared by using cultivation-dependent and cultivation-independent methods. For cultivation-dependent method, Random amplified polymorphic DNA (RAPD) profiles revealed that the bacterial genetic diversity of V. radiata and A. mangium rhizospheres was higher than that of A. hypogaea rhizosphere. For cultivation-independent method, Denaturing gradient gel electrophoresis (DGGE) profiles of PCR-amplified 16S rRNA genes revealed the difference in bacterial community and diversity of rhizospheres collected from 3 legumes. The ribotype richness which indicates species diversity, was highest in V. radiata rhizosphere, followed by A. hypogaea and A. mangium rhizospheres, respectively. Three kinds of media were used to cultivate different target groups of bacteria. The result indicates that the communities of cultivable bacteria in 3 rhizospheres recovered from nutrient agar (NA) medium were mostly different from each other, while Bradyrhizobium selective medium (BJSM) and nitrogen-free medium shaped the communities of cultivable bacteria. Nine isolates grown on BJSM were identified by 16S rRNA gene sequence analysis. These isolates were very closely related (with 96% to 99% identities) to either one of the three groups including Cupriavidus-Ralstonia group, Bacillus group and Bradyrhizobium-Bosea-Afipia group. The rhizospheres were also examined for their enzymatic patterns. Of 19 enzymes tested, 3 rhizospheres were distinguishable by the presence or the absence of leucine acrylamidase and acid phosphatase. The selected cultivable bacteria recovered from NA varied in their abilities to produce indole-acetic acid and ammnonia. The resistance to 10 antibiotics was indistinguishable among bacteria isolated from different rhizospheres. PMID:22806857

  16. Family acholeplasmataceae (including phytoplasmas)

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The family Acholeplasmataceae was originally established to accommodate the genus Acholeplasma, comprising the mollicutes that could be cultivated without the supplement of cholesterol and that use UGA as a stop codon instead of coding for tryptophan. It was later shown that the phytoplasmas, a larg...

  17. Investigation of Methanogen Diversity in Stored Swine Manure

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Consolidated storage of swine manure is associated with the microbial production of a variety of odors and emissions, including ammonia, organic acids, alcohols, and hydrogen sulfide. Large quantities of methane are also produced from such facilities. In the United States, methane emissions from l...

  18. KINETICS OF ETHANOL BIODEGRADATION UNDER METHANOGENIC CONDITIONS IN GASOLINE SPILLS

    EPA Science Inventory

    Ethanol is commonly used as a fuel oxygenate. A concern has been raised that biodegradation of ethanol from a spill of gasoline may inhibit the natural biodegradation of fuel hydrocarbons, including benzene. Ethanol is miscible in water, and ethanol is readily metabolized by mi...

  19. Role of methanogens and other bacteria in degradation of dimethyl sulfide and methanethiol in anoxic freshwater sediments

    SciTech Connect

    Lomans, B.P.; Op den Camp, H.J.M.; Pol, A.; Drift, C. van der; Vogels, G.D.

    1999-05-01

    The roles of several trophic groups of organisms (methanogens and sulfate- and nitrate-reducing bacteria) in the microbial degradation of methanethiol (MT) and dimethyl sulfide (DMS) were studied in freshwater sediments. The incubation of DMS- and MT-amended slurries revealed that methanogens are the dominant DMS and MT utilizers in sulfate-poor freshwater systems. In sediment slurries, which were depleted of sulfate, 75 {micro}mol of DMS was stoichiometrically converted into 112 {micro}mol of methane. The addition of methanol or MT to DMS-degrading slurries at concentrations similar to that of DMS reduced DMS degradation rates. This indicates that the methanogens in freshwater sediments, which degrade DMS, are also consumers of methanol and MT. To verify whether a competition between sulfate-reducing and methanogenic bacteria for DMS or MT takes place in sulfate-rich freshwater systems, the effects of sulfate and inhibitors, like bromoethanesulfonic acid, molybdate, and tungstate, on the degradation of MT and DMS were studied. The results for these sulfate-rich and sulfate-amended slurry incubations clearly demonstrated that besides methanogens, sulfate-reducing bacteria take part in MT and DMS degradation in freshwater sediments, provided that sulfate is available. The possible involvement of an interspecies hydrogen transfer in these processes is discussed. In general, the study provides evidence for methanogenesis as a major sink for MT and DMS in freshwater sediments.

  20. Classes of organic molecules targeted by a methanogenic microbial consortium grown on sedimentary rocks of various maturities

    PubMed Central

    Meslé, Margaux; Dromart, Gilles; Haeseler, Frank; Oger, Philippe M.

    2015-01-01

    Organic-rich shales are populated by methanogenic consortia that are able to degrade the fossilized organic matter into methane gas. To identify the organic fraction effectively degraded, we have sequentially depleted two types of organic-rich sedimentary rocks, shale, and coal, at two different maturities, by successive solvent extractions to remove the most soluble fractions (maltenes and asphaltenes) and isolate kerogen. We show the ability of the consortia to produce methane from all rock samples, including those containing the most refractory organic matter, i.e., the kerogen. Shales yielded higher methane production than lignite and coal. Mature rocks yielded more methane than immature rocks. Surprisingly, the efficiency of the consortia was not influenced by the removal of the easily biodegradable fractions contained in the maltenes and asphaltenes. This suggests that one of the limitations of organic matter degradation in situ may be the accessibility to the carbon and energy source. Indeed, bitumen has a colloidal structure that may prevent the microbial consortia from reaching the asphaltenes in the bulk rock. Solvent extractions might favor the access to asphaltenes and kerogen by modifying the spatial organization of the molecules in the rock matrix. PMID:26136731

  1. Classes of organic molecules targeted by a methanogenic microbial consortium grown on sedimentary rocks of various maturities.

    PubMed

    Meslé, Margaux; Dromart, Gilles; Haeseler, Frank; Oger, Philippe M

    2015-01-01

    Organic-rich shales are populated by methanogenic consortia that are able to degrade the fossilized organic matter into methane gas. To identify the organic fraction effectively degraded, we have sequentially depleted two types of organic-rich sedimentary rocks, shale, and coal, at two different maturities, by successive solvent extractions to remove the most soluble fractions (maltenes and asphaltenes) and isolate kerogen. We show the ability of the consortia to produce methane from all rock samples, including those containing the most refractory organic matter, i.e., the kerogen. Shales yielded higher methane production than lignite and coal. Mature rocks yielded more methane than immature rocks. Surprisingly, the efficiency of the consortia was not influenced by the removal of the easily biodegradable fractions contained in the maltenes and asphaltenes. This suggests that one of the limitations of organic matter degradation in situ may be the accessibility to the carbon and energy source. Indeed, bitumen has a colloidal structure that may prevent the microbial consortia from reaching the asphaltenes in the bulk rock. Solvent extractions might favor the access to asphaltenes and kerogen by modifying the spatial organization of the molecules in the rock matrix. PMID:26136731

  2. Integrating cultivation history into EBIPM

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Ecologically based invasive plant management (EBIPM) is a systematic thinking and planning process to assist with applying the appropriate combination of tools and strategies to addrress the underlying cause of invasion rather than simply controlling invasive annual grass abundance. Cultivation his...

  3. Cultivating Spontaneous Self-Discipline.

    ERIC Educational Resources Information Center

    O'Shaughnessy, Molly

    1998-01-01

    Draws on contemporary sources to provide strategies for cultivating self-discipline. Advocates self-healing for the adult to be free from destructive attitudes and personal history that can keep adults from being mindful of the child's needs, perspective, and potential. Concludes with ways to facilitate a truly Montessori approach to discipline.…

  4. Cultivating Spiritual Reflectivity in Teachers.

    ERIC Educational Resources Information Center

    Mayes, Clifford

    2001-01-01

    Discusses the need to address spiritual beliefs in teacher education, arguing that: because spiritual impulses are so primary for many people, they must be addressed in teacher education; educators must recognize and cultivate students' spiritual motivations to teach and foster reflectivity; and student teachers must learn what they can legally…

  5. Plasmid DNA from the acetotrophic methanogen Methanosarcina acetivorans

    SciTech Connect

    Sowers, K.R.; Gunsalus, R.P. )

    1988-10-01

    Nine acetotrophic and three methylotrophic strains of methane-producing bacteria were screened for the presence of plasmid DNA. Plasmids were detected in three marine isolates, including Methanosarcina acetivorans. All three plasmids appeared to be similar based on size and restriction site analyses. The plasmid from M. acetivorans, designated pC2A, was approximately 5.1 kilobase pairs in size and was estimated to be present in a low copy number of six plasmids per genome. Multimers were also observed. A restriction map was constructed. The function of this plasmid is cryptic.

  6. High-sulfate, high-chemical oxygen demand wastewater treatment using aerated methanogenic fluidized beds

    SciTech Connect

    Zitomer, D.H.; Shrout, J.D.

    2000-02-01

    Many industrial wastewaters have both high organic pollution and sulfate (SO{sub 4}{sup {minus}2}) concentrations. Although biological conversion of organics to methane may be an economical chemical oxygen demand (COD) removal option, significant inhibition of methane production results from reduction of SO{sub 4}{sup {minus}2} to hydrogen sulfide (H{sub 2}S), which is inhibitory to methanogenic microorganisms. Therefore, sulfate-containing wastewater is often not amenable to conventional anaerobic treatment. Recently, limited aeration of recycle flow to hybrid and baffled reactors has been used to treat this wastewater and has been shown to reduce aqueous H{sub 2}S concentrations by causing production of uninhibitory sulfur (S{degree}) and thiosulfate (S{sub 2}O{sub 3}{sup {minus}2}) as well as gas stripping volatile H{sub 2}S. In this study, directly aerated methanogenic fluidized bed reactors (FBRs) achieved increased methane production compared to strictly anaerobic FBRs treating high-sulfate wastewater. Oxygen transfer satisfying up to 28% of the COD load resulted in maximum specific oxygen utilization rates of 0.20 mg oxygen/g volatile solids{center{underscore}dot}min, with significant, concomitant methane production. Under typically inhibitory SO{sub 4}{sup {minus}2} loading, higher aeration caused increased effluent SO{sub 4}{sup {minus}2}, increased H{sub 2}S mass in the offgas, and lower reactor H{sub 2}S concentration. As a result, COD removal increased from 25% for a strictly anaerobic FBR to 87% for an aerated FBR. In addition, aerated systems required significantly less alkalinity supplementation to maintain a pH value of 7, ostensibly because of stripping of acidic carbon dioxide. The potential pH increase associated with aeration also shifts sulfide speciation to less toxic disulfide. Direct, limited aeration of methanogenic FBRs is described as a method for increased COD removal when treating high-COD, high-sulfate wastewater.

  7. NMR studies of osmoregulation in methanogenic archaebacteria. [NMR (nuclear magnetic resonance)

    SciTech Connect

    Robertson, D.E.

    1991-01-01

    Methanogens are strict anaerobic archaebacteria whose metabolism centers around the reduction of CO[sub 2] to CH[sub 4]. Their environments are often extreme (high temperatures, high salt, few nutrients, etc.) and they may have evolved unique ways to handle these stresses. It is proposed that methanogenic archaebacteria respond to osmotic stress by accumulating a series of organic solutes. In two strains of marine methanogens, Methanogenium cariaci and Methanococcus thermolithotrophicus, four key organic solutes are observed: L-[alpha]-glutamate, [beta]-glutamate, N[sup e]-acetyl-[beta]-lysine, and glycine betaine. The first three of these are synthesized de novo; glycine betaine is transported into the Mg. cariaci cells from the medium. In the absence of betaine, Mg. cariaci synthesizes N[sup e]-acetyl-[beta]-lysine as the dominant osmolyte. Mc. thermolithotrophicus also synthesizes N[sup e]-acetyl-[beta]-lysine but only at salt concentrations greater than 1 M. In Mc. thermolithotrophicus intracellular potassium ion concentrations, determined by [sup 39]K NMR spectroscopy, are balanced by the total concentration of anionic amino acid species, [alpha]-glutamate and [beta]-glutamate. Turnover of the organic solutes has been monitored using [sup 13]C-pulse/[sup 12]C-chase, and [sup 15]N-pulse/[sup 14]N-chase experiments. The [beta]-amino acids exhibit slower turnover rates compared to L-[alpha]-glutamate or aspartate, consistent with their role as compatible solutes. Biosynthetic information for the [beta]-amino acids was provided by [sup 13]C-label incorporation and steady state labeling experiments. [beta]-glutamate shows a lag in [sup 13]C uptake from [sup 13]CO[sub 2], indicative of its biosynthesis from a precursor not in equilibrium with the soluble L-[alpha]-glutamate pool, probably a macromolecule. A novel biosynthetic pathway is proposed for N[sup e]-acetyl-[beta]-lysine from the diaminopimelate pathway.

  8. Oceanic nickel depletion and a methanogen famine before the Great Oxidation Event.

    PubMed

    Konhauser, Kurt O; Pecoits, Ernesto; Lalonde, Stefan V; Papineau, Dominic; Nisbet, Euan G; Barley, Mark E; Arndt, Nicholas T; Zahnle, Kevin; Kamber, Balz S

    2009-04-01

    It has been suggested that a decrease in atmospheric methane levels triggered the progressive rise of atmospheric oxygen, the so-called Great Oxidation Event, about 2.4 Gyr ago. Oxidative weathering of terrestrial sulphides, increased oceanic sulphate, and the ecological success of sulphate-reducing microorganisms over methanogens has been proposed as a possible cause for the methane collapse, but this explanation is difficult to reconcile with the rock record. Banded iron formations preserve a history of Precambrian oceanic elemental abundance and can provide insights into our understanding of early microbial life and its influence on the evolution of the Earth system. Here we report a decline in the molar nickel to iron ratio recorded in banded iron formations about 2.7 Gyr ago, which we attribute to a reduced flux of nickel to the oceans, a consequence of cooling upper-mantle temperatures and decreased eruption of nickel-rich ultramafic rocks at the time. We measured nickel partition coefficients between simulated Precambrian sea water and diverse iron hydroxides, and subsequently determined that dissolved nickel concentrations may have reached approximately 400 nM throughout much of the Archaean eon, but dropped below approximately 200 nM by 2.5 Gyr ago and to modern day values ( approximately 9 nM) by approximately 550 Myr ago. Nickel is a key metal cofactor in several enzymes of methanogens and we propose that its decline would have stifled their activity in the ancient oceans and disrupted the supply of biogenic methane. A decline in biogenic methane production therefore could have occurred before increasing environmental oxygenation and not necessarily be related to it. The enzymatic reliance of methanogens on a diminishing supply of volcanic nickel links mantle evolution to the redox state of the atmosphere. PMID:19360085

  9. Peat: home to novel syntrophic species that feed acetate- and hydrogen-scavenging methanogens.

    PubMed

    Schmidt, Oliver; Hink, Linda; Horn, Marcus A; Drake, Harold L

    2016-08-01

    Syntrophic bacteria drive the anaerobic degradation of certain fermentation products (e.g., butyrate, ethanol, propionate) to intermediary substrates (e.g., H2, formate, acetate) that yield methane at the ecosystem level. However, little is known about the in situ activities and identities of these syntrophs in peatlands, ecosystems that produce significant quantities of methane. The consumption of butyrate, ethanol or propionate by anoxic peat slurries at 5 and 15 °C yielded methane and CO2 as the sole accumulating products, indicating that the intermediates H2, formate and acetate were scavenged effectively by syntrophic methanogenic consortia. 16S rRNA stable isotope probing identified novel species/strains of Pelobacter and Syntrophomonas that syntrophically oxidized ethanol and butyrate, respectively. Propionate was syntrophically oxidized by novel species of Syntrophobacter and Smithella, genera that use different propionate-oxidizing pathways. Taxa not known for a syntrophic metabolism may have been involved in the oxidation of butyrate (Telmatospirillum-related) and propionate (unclassified Bacteroidetes and unclassified Fibrobacteres). Gibbs free energies (ΔGs) for syntrophic oxidations of ethanol and butyrate were more favorable than ΔGs for syntrophic oxidation of propionate. As a result of the thermodynamic constraints, acetate transiently accumulated in ethanol and butyrate treatments but not in propionate treatments. Aceticlastic methanogens (Methanosarcina, Methanosaeta) appeared to outnumber hydrogenotrophic methanogens (Methanocella, Methanoregula), reinforcing the likely importance of aceticlastic methanogenesis to the overall production of methane. ΔGs for acetogenesis from H2 to CO2 approximated to -20 kJ mol(-1) when acetate concentrations were low, indicating that acetogens may have contributed to the flow of carbon and reductant towards methane. PMID:26771931

  10. Analysis of methanogenic and methanotrophic activity at the western margin of the Greenland Ice Sheet

    NASA Astrophysics Data System (ADS)

    Broemsen, E. L.; Webster, K. D.; Dieser, M.; Pratt, L. M.; Christner, B. C.

    2012-12-01

    Anoxic conditions in environments beneath the world's glaciers and ice sheets provide plausible habitats supporting the microbial production of methane. Recent reports of potential methane sources beneath the Greenland Ice Sheet (GrIS) suggest in situ production by an active community of methanogens. Beneath the GrIS, microbially derived methane can be dissolved in subglacial water, and during periods of melting, can exchange with the atmosphere at sites of subglacial discharge. Transfer of methane from subglacial fluids to the atmosphere could be a significant climate factor, but few data are available to make such assessments. The specific aim of this study was to characterize the composition and activity of methanogens and methanotrophs present in samples of subglacial outflow at the ice sheet margin near Kangerlussuaq, Greenland. Subglaical water was collected twice-weekly over a nine week period (mid July to mid September of 2012) and the dissolved methane concentration in the samples was determined via gas chromatography. Extracted RNA and DNA from the subglacial water was analyzed by analysis of 16s rRNA and rRNA genes present in the subglacial assemblages. From the molecular results we infer the presence of active methanogens related to the order Methanosarcinales. Further, locally elevated concentrations of atmospheric methane as high as 1.92 ± 0.03 ppmv, were detected in the ice tunnel of the subglacial outflow using open-path laser spectrometry. From these data we estimate rates of methane release at the ice sheet margin during the summer melt months at this geographical location. The results provide a context for addressing the impact that deglaciation will have on the release of greenhouse gases from ice sheets on a warming Earth.

  11. Methanotrophic and Methanogenic Communities in Swiss Alpine Fens Dominated by Carex rostrata and Eriophorum angustifolium

    PubMed Central

    Cheema, Simrita; Henneberger, Ruth

    2015-01-01

    Vascular plants play a key role in controlling CH4 emissions from natural wetlands, because they influence CH4 production, oxidation, and transport to the atmosphere. Here we investigated differences in the abundance and composition of methanotrophic and methanogenic communities in three Swiss alpine fens dominated by different vascular plant species under natural conditions. The sampling locations either were situated at geographically distinct sites with different physicochemical properties but the same dominant plant species (Carex rostrata) or were located within the same site, showing comparable physicochemical pore water properties, but had different plant species (C. rostrata or Eriophorum angustifolium). All three locations were permanently submerged and showed high levels of CH4 emissions (80.3 to 184.4 mg CH4 m−2 day−1). Soil samples were collected from three different depths with different pore water CH4 and O2 concentrations and were analyzed for pmoA and mcrA gene and transcript abundance and community composition, as well as soil structure. The dominant plant species appeared to have a significant influence on the composition of the active methanotrophic communities (transcript level), while the methanogenic communities differed significantly only at the gene level. Yet no plant species-specific microbial taxa were discerned. Moreover, for all communities, differences in composition were more pronounced with the site (i.e., with different physicochemical properties) than with the plant species. Moreover, depth significantly influenced the composition of the active methanotrophic communities. Differences in abundance were generally low, and active methanotrophs and methanogens coexisted at all three locations and depths independently of CH4 and O2 concentrations or plant species. PMID:26092454

  12. Biogas production and methanogenic archaeal community in mesophilic and thermophilic anaerobic co-digestion processes.

    PubMed

    Yu, D; Kurola, J M; Lähde, K; Kymäläinen, M; Sinkkonen, A; Romantschuk, M

    2014-10-01

    Over 258 Mt of solid waste are generated annually in Europe, a large fraction of which is biowaste. Sewage sludge is another major waste fraction. In this study, biowaste and sewage sludge were co-digested in an anaerobic digestion reactor (30% and 70% of total wet weight, respectively). The purpose was to investigate the biogas production and methanogenic archaeal community composition in the anaerobic digestion reactor under meso- (35-37 °C) and thermophilic (55-57 °C) processes and an increasing organic loading rate (OLR, 1-10 kg VS m(-3) d(-1)), and also to find a feasible compromise between waste treatment capacity and biogas production without causing process instability. In summary, more biogas was produced with all OLRs by the thermophilic process. Both processes showed a limited diversity of the methanogenic archaeal community which was dominated by Methanobacteriales and Methanosarcinales (e.g. Methanosarcina) in both meso- and thermophilic processes. Methanothermobacter was detected as an additional dominant genus in the thermophilic process. In addition to operating temperatures, the OLRs, the acetate concentration, and the presence of key substrates like propionate also affected the methanogenic archaeal community composition. A bacterial cell count 6.25 times higher than archaeal cell count was observed throughout the thermophilic process, while the cell count ratio varied between 0.2 and 8.5 in the mesophilic process. This suggests that the thermophilic process is more stable, but also that the relative abundance between bacteria and archaea can vary without seriously affecting biogas production. PMID:24837280

  13. Response of the methanogenic microbial communities in Amazonian oxbow lake sediments to desiccation stress.

    PubMed

    Conrad, Ralf; Ji, Yang; Noll, Matthias; Klose, Melanie; Claus, Peter; Enrich-Prast, Alex

    2014-06-01

    Methanogenic microbial communities in soil and sediment function only when the environment is inundated and anoxic. In contrast to submerged soils, desiccation of lake sediments happens only rarely. However, some predictions suggest that extreme events of drying will become more common in the Amazon region, and this will promote an increase in sediments drying and exposure. We asked whether and how such methanogenic communities can withstand desiccation stress. Therefore, we determined the rates and pathways of CH(4) production (analysis of CH(4) and δ(13) C of CH(4), CO(2) and acetate), the copy numbers of bacterial and archaeal 16S rRNA genes and mcrA genes (quantitative PCR), and the community composition of Archaea and Bacteria (T-RFLP and pyrosequencing) in oxbow lake sediments of rivers in the Brazilian Amazon region. The rivers were of white water, black water and clear water type. The measurements were done with sediment in fresh state and after drying and rewetting. After desiccation and rewetting the composition of both, the archaeal and bacterial community changed. Since lake sediments from white water rivers exhibited only negligible methanogenic activity, probably because of relatively high iron and low organic matter content, they were not further analysed. The other sediments produced CH(4), with hydrogenotrophic methanogenesis usually accounting for > 50% of total activity. After desiccation and rewetting, archaeal and bacterial gene copy numbers decreased. The bacterial community showed a remarkable increase of Clostridiales from about 10% to > 30% of all Bacteria, partially caused by proliferation of specific taxa as the numbers of OTU shared with fresh sediment decreased from about 9% to 3%. Among the Archaea, desiccation specifically enhanced the relative abundance of either Methanocellales (black water) and/or Methanosarcinaceae (clear water). Despite the changes in gene copy numbers and composition of the microbial community, rates of CH

  14. Detection of methanogenic archaea in the pitchers of the Northern pitcher plant (Sarracenia purpurea ).

    PubMed

    Krieger, Joseph R; Kourtev, Peter S

    2012-02-01

    Carnivorous plants of the genus Sarracenia rely on microorganisms in their pitchers to decompose drowned insects. The environment inside pitchers is considered to be aerobic; however, there might be zones, such as at the bottom of the pitcher, where anaerobic conditions develop. Samples of the sediment at the bottom of Sarracenia purpurea pitchers were analyzed for the presence of archaea, using PCR and sequencing of the 16S rRNA gene. Archaeal DNA was detected in 20% of sampled pitchers. All sequences were closely related to Methanobrevibacter . Therefore, pitchers may contain anoxic zones inhabited by methanogens. PMID:22263906

  15. Investigation of methanogenic community structures in rural biogas digesters from different climatic regions in Yunnan, southwest China.

    PubMed

    Dong, Minghua; Wu, Yan; Li, Qiumin; Tian, Guangliang; Yang, Bin; Li, Yingjuan; Zhang, Lijuan; Wang, Yongxia; Xiao, Wei; Yin, Fang; Zhao, Xingling; Zhang, Wudi; Cui, Xiaolong

    2015-05-01

    Understanding of the microbial community structures of the biogas digesters in different climatic regions can help improve the methane production in the fermentation process. The methanogenic archaeal diversity in four rural biogas digesters (BNA, JSA, LJA, and XGA) was investigated by a culture-independent rRNA approach in different climatic regions in Yunnan. Community structure composed of 711 clones in the all libraries. A total of 33 operational taxonomic units (OTUs) were detected, and major groups of methanogens were the orders Methanosarcinales and Methanomicrobiales. 63.2 % of all archaeal OTUs belong to the order Methanosarcinales which mostly contain acetotrophic methanogens. Methanomicrobiales (19.5 % in all OTUs) were detected in considerable number. Additionally, there were minor rates of uncultured archaea. The principal component analysis indicated that the genus Methanosaeta was mainly affected by the fermentation temperatures. PMID:25575901

  16. Anaerobic 2-propanol degradation in anoxic paddy soil and the possible role of methanogens in its degradation.

    PubMed

    Tonouchi, Akio

    2004-08-01

    The anaerobic degradation of 2-propanol in anoxic paddy soil was studied with soil cultures and a 2-propanol-utilizing methanogen. Acetone was the first and the major intermediate involved in the methanogenic degradation of 2-propanol. Analyses with a methanogenesis inhibitor, bacteria antibiotics, and the addition of H2 to the gas phase revealed that 2-propanol oxidation to acetone directly occurred using 2-propanol-utilizing methanogens, but not with H2-producing syntrophic bacteria, for which the removal of acetone is required for complete 2-propanol oxidation. The 2-propanol-utilizing strain IIE1, which is phylogenetically closely related to Methanoculleus palmolei, was isolated from paddy soil, and the potential role of the strain in 2-propanol degradation was investigated. 2-Propanol is one of the representative fermentation intermediates in anaerobic environments. This is the first report on the anaerobic 2-propanol degradation process. PMID:15297909

  17. Methanol induces low temperature resilient methanogens and improves methane generation from domestic wastewater at low to moderate temperatures.

    PubMed

    Saha, Shaswati; Badhe, Neha; De Vrieze, Jo; Biswas, Rima; Nandy, Tapas

    2015-01-01

    Low temperature (<20 °C) limits bio-methanation of sewage. Literature shows that hydrogenotrophic methanogens can adapt themselves to low temperature and methanol is a preferred substrate by methanogens in cold habitats. The study hypothesizes that methanol can induce the growth of low-temperature resilient, methanol utilizing, hydrogenotrophs in UASB reactor. The hypothesis was tested in field conditions to evaluate the impact of seasonal temperature variations on methane yield in the presence and absence of methanol. Results show that 0.04% (v/v) methanol increased methane up to 15 times and its effect was more pronounced at lower temperatures. The qPCR analysis showed the presence of Methanobacteriales along with Methanosetaceae in large numbers. This indicates methanol induced the growth of both the hydrogenotrophic and acetoclastic groups through direct and indirect routes, respectively. This study thus demonstrated that methanol can impart resistance in methanogenic biomass to low temperature and can improve performance of UASB reactor. PMID:25913884

  18. Hydrogen Limitation and Syntrophic Growth among Natural Assemblages of Thermophilic Methanogens at Deep-sea Hydrothermal Vents

    PubMed Central

    Topçuoğlu, Begüm D.; Stewart, Lucy C.; Morrison, Hilary G.; Butterfield, David A.; Huber, Julie A.; Holden, James F.

    2016-01-01

    Thermophilic methanogens are common autotrophs at hydrothermal vents, but their growth constraints and dependence on H2 syntrophy in situ are poorly understood. Between 2012 and 2015, methanogens and H2-producing heterotrophs were detected by growth at 80°C and 55°C at most diffuse (7–40°C) hydrothermal vent sites at Axial Seamount. Microcosm incubations of diffuse hydrothermal fluids at 80°C and 55°C demonstrated that growth of thermophilic and hyperthermophilic methanogens is primarily limited by H2 availability. Amendment of microcosms with NH4+ generally had no effect on CH4 production. However, annual variations in abundance and CH4 production were observed in relation to the eruption cycle of the seamount. Microcosm incubations of hydrothermal fluids at 80°C and 55°C supplemented with tryptone and no added H2 showed CH4 production indicating the capacity in situ for methanogenic H2 syntrophy. 16S rRNA genes were found in 80°C microcosms from H2-producing archaea and H2-consuming methanogens, but not for any bacteria. In 55°C microcosms, sequences were found from H2-producing bacteria and H2-consuming methanogens and sulfate-reducing bacteria. A co-culture of representative organisms showed that Thermococcus paralvinellae supported the syntrophic growth of Methanocaldococcus bathoardescens at 82°C and Methanothermococcus sp. strain BW11 at 60°C. The results demonstrate that modeling of subseafloor methanogenesis should focus primarily on H2 availability and temperature, and that thermophilic H2 syntrophy can support methanogenesis within natural microbial assemblages and may be an important energy source for thermophilic autotrophs in marine geothermal environments. PMID:27547206

  19. Reactive transport model of growth and methane production by high-temperature methanogens in hydrothermal regions of the subseafloor

    NASA Astrophysics Data System (ADS)

    Stewart, L. C.; Algar, C. K.; Topçuoğlu, B. D.; Fortunato, C. S.; Larson, B. I.; Proskurowski, G. K.; Butterfield, D. A.; Vallino, J. J.; Huber, J. A.; Holden, J. F.

    2014-12-01

    Hydrogenotrophic methanogens are keystone high-temperature autotrophs in deep-sea hydrothermal vents and tracers of habitability and biogeochemical activity in the hydrothermally active subseafloor. At Axial Seamount, nearly all thermophilic methanogens are Methanothermococcus and Methanocaldococcus species, making this site amenable to modeling through pure culture laboratory experiments coupled with field studies. Based on field microcosm incubations with 1.2 mM, 20 μM, or no hydrogen, the growth of methanogens at 55°C and 80°C is limited primarily by temperature and hydrogen availability, with ammonium amendment showing no consistent effect on total methane output. The Arrhenius constants for methane production by Methanocaldococcus jannaschii (optimum 82°C) and Methanothermococcus thermolithotrophicus (optimum 65°C) were determined in pure culture bottle experiments. The Monod constants for hydrogen concentration were measured by growing both organisms in a 2-liter chemostat at two dilution rates; 55°C, 65°C and 82°C; and variable hydrogen concentrations. M. jannaschii showed higher ks and Vmax constants than M. thermolithotrophicus. In the field, hydrogen and methane concentrations in hydrothermal end-member and low-temperature diffuse fluids were measured, and the concentrations of methanogens that grow at 55°C and 80°C in diffuse fluids were determined using most-probable-number estimates. Methane concentration anomalies in diffuse fluids relative to end-member hydrothermal concentrations and methanogen cell concentrations are being used to constrain a 1-D reactive transport model using the laboratory-determined Arrhenius and Monod constants for methane production by these organisms. By varying flow path length and subseafloor cell concentrations in the model, our goal is to determine solutions for the potential depth of the subseafloor biosphere coupled with the amount of methanogenic biomass it contains.

  20. Application of stable isotope measurements and microbiological analysis for detecting methanogenic activity in a temperate forest wetland

    NASA Astrophysics Data System (ADS)

    Itoh, M.; Katsuyama, C.; Kondo, N.; Ohte, N.; Kato, K.

    2009-12-01

    Generally, forest soils act as a sink for methane (CH4). However, wetlands in riparian zones are recently reported to be “hot spots” of CH4 emissions, especially in forests under a humid climate. To understand how environmental conditions (i.e. hydrological and/or geomorphic condition) control on CH4 production, we investigated both methanogenic pathways (CO2/H2 reduction and acetate fermentation) and metahanogenic microbial communities in a wetland in a temperate forest catchment, central Japan. We used stable carbon isotopic analysis for detecting change in methanogenic pathways, and applied microbiological analysis for understanding the structure of methanogenic community. CH4 emission rates in wetland were strongly dependent on soil temperatures, and were highest in summer and lowest in winter. δ13CO2 increased with CH4 production in every summer, suggesting preferential use of 12CO2 as substrate for CO2/H2 reduction methanogenesis during high CH4 production period. δ13CH4 also increased in summer with δ13CO2. δ13CH4 changed more wildly than δ13CO2 did in summer with normal precipitation when CH4 production was strongly activated under high temperature and high groundwater table condition. This indicates increase in acetoclastic methanogenesis under hot and wet condition, considering that acetclastic methnogens produce heavier CH4 than that from CO2/H2 reducing pathway. Methanogen community composition estimated by cloning and sequence analyses implied that both acetoclastic and CO2/H2 reducing methanogens prevailed in wetland soil sampled in summer. This was consistent with the results of isotope measuremaents. Our results contribute to understand fully how the CH4 production changes with environmental conditions, with considering the activities of both main methanogenic pathway (from CO2 and acetate).

  1. Hydrogen Limitation and Syntrophic Growth among Natural Assemblages of Thermophilic Methanogens at Deep-sea Hydrothermal Vents.

    PubMed

    Topçuoğlu, Begüm D; Stewart, Lucy C; Morrison, Hilary G; Butterfield, David A; Huber, Julie A; Holden, James F

    2016-01-01

    Thermophilic methanogens are common autotrophs at hydrothermal vents, but their growth constraints and dependence on H2 syntrophy in situ are poorly understood. Between 2012 and 2015, methanogens and H2-producing heterotrophs were detected by growth at 80°C and 55°C at most diffuse (7-40°C) hydrothermal vent sites at Axial Seamount. Microcosm incubations of diffuse hydrothermal fluids at 80°C and 55°C demonstrated that growth of thermophilic and hyperthermophilic methanogens is primarily limited by H2 availability. Amendment of microcosms with NH4 (+) generally had no effect on CH4 production. However, annual variations in abundance and CH4 production were observed in relation to the eruption cycle of the seamount. Microcosm incubations of hydrothermal fluids at 80°C and 55°C supplemented with tryptone and no added H2 showed CH4 production indicating the capacity in situ for methanogenic H2 syntrophy. 16S rRNA genes were found in 80°C microcosms from H2-producing archaea and H2-consuming methanogens, but not for any bacteria. In 55°C microcosms, sequences were found from H2-producing bacteria and H2-consuming methanogens and sulfate-reducing bacteria. A co-culture of representative organisms showed that Thermococcus paralvinellae supported the syntrophic growth of Methanocaldococcus bathoardescens at 82°C and Methanothermococcus sp. strain BW11 at 60°C. The results demonstrate that modeling of subseafloor methanogenesis should focus primarily on H2 availability and temperature, and that thermophilic H2 syntrophy can support methanogenesis within natural microbial assemblages and may be an important energy source for thermophilic autotrophs in marine geothermal environments. PMID:27547206

  2. Characterization of a Methanogenic Community within an Algal Fed Anaerobic Digester

    PubMed Central

    Ellis, Joshua T.; Tramp, Cody; Sims, Ronald C.; Miller, Charles D.

    2012-01-01

    The microbial diversity and metabolic potential of a methanogenic consortium residing in a 3785-liter anaerobic digester, fed with wastewater algae, was analyzed using 454 pyrosequencing technology. DNA was extracted from anaerobic sludge material and used in metagenomic analysis through PCR amplification of the methyl-coenzyme M reductase α subunit (mcrA) gene using primer sets ML, MCR, and ME. The majority of annotated mcrA sequences were assigned taxonomically to the genera Methanosaeta in the order Methanosarcinales. Methanogens from the genus Methanosaeta are obligate acetotrophs, suggesting this genus plays a dominant role in methane production from the analyzed fermentation sample. Numerous analyzed sequences within the algae fed anaerobic digester were unclassified and could not be assigned taxonomically. Relative amplicon frequencies were determined for each primer set to determine the utility of each in pyrosequencing. Primer sets ML and MCR performed better quantitatively (representing the large majority of analyzed sequences) than primer set ME. However, each of these primer sets was shown to provide a quantitatively unique community structure, and thus they are of equal importance in mcrA metagenomic analysis. PMID:23724331

  3. Effect of humic acid in leachate on specific methanogenic activity of anaerobic granular sludge.

    PubMed

    Guo, Mengfei; Xian, Ping; Yang, Longhui; Liu, Xi; Zhan, Longhui; Bu, Guanghui

    2015-01-01

    In order to find out the effects of humic acid (HA) in anaerobic-treated landfill leachate on granular sludge, the anaerobic biodegradability of HA as well as the influences of HA on the total cumulative methane production, the anaerobic methanization process and the specific methanogenic activity (SMA) of granular sludge are studied in this paper. Experimental results show that as a non-biodegradable organic pollutant, HA is also difficult to be decomposed by microbes in the anaerobic reaction process. Presence of HA and changes in the concentration have no significant influences on the total cumulative methane production and the anaerobic methanization process of granular sludge. Besides, the total cumulative methane production cannot reflect the inhibition of toxics on the methanogenic activity of granular sludge on the premise of sufficient reaction time. Results also show that HA plays a promoting role on SMA of granular sludge. Without buffering agent the SMA value increased by 19.2% on average due to the buffering and regulating ability of HA, while with buffering agent the SMA value increased by 5.4% on average due to the retaining effect of HA on the morphology of the sludge particles. However, in the presence of leachate the SMA value decreased by 27.6% on average, because the toxic effect of the toxics in the leachate on granular sludge is much larger than the promoting effect of HA. PMID:25915186

  4. Lack of mutagenicity of methanogenic digester effluent in the Salmonella/microsome test

    SciTech Connect

    Wei, C.; Oblinger, J.L.; Richter, M.F.

    1984-05-01

    Kaplan Industries (Bartow, FL) has inititated a large-scale waste utilization program to dispose 500 tons/day of manure from a 20,000 head cattle feed lot. About 30% of the solid material in the manure is separated and recycled as an organic fertilizer. The liquid portion, with some suspended solids, is transferred to methane digesters. Methanogenic digester effluent is centrifuged to yield a 25% protein product that can be used as an animal feed supplement; the liquid effluent is used for agricultural irrigation. Studies on the use of municipal sewage sludge as fertilizer on farmlands have demonstrated that high levels of PCB's have been found in crops grown on PCB-contaminated sludge-amended soils. Since similar reaction products, or their metabolites may be produced from heavy metals, pesticide residues, antibiotics or residues from feed present in animal manure during anaerobic conversion of this biomass to methane, better assessment of the potential hazard of digester effluent is needed. The Ames Salmonella/microsome assay for mutagenicity has been widely applied in in vitro tests for potential carcinogenicity and mutagenicity of environmental chemicals or mixtures. In this study they used the three most sensitive strains of Salmonella typhimurium, i.e., TA 97, TA 98 and TA 100, to evaluate the possible presence of genotoxic compounds in the methanogenic digester effluent.

  5. Molecular biology and genetics of the acetate-utilizing methanogenic bacteria

    SciTech Connect

    Gunsalus, R.P.

    1991-01-01

    Acetate conversion to methane and C0{sub 2} by the methanogenic archaebacteria is a primary rate limiting step in anaerobic biodegradative processes in nature. However, the genetic study of these organisms has not been experimentally tractable due to the inability to grow and plate the organisms as single cells, and to extract high molecular weight DNA and RNA without shearing. The acetate-utilizing species, Methanosarcina thermolphila TM-1, is being used for the proposed genetic and molecular studies because, unlike previously described acetotrophic methanosarcina that have a thick heteropolysaccharide cell wall, this species can be cultured in a unicellular form that has a protein cell wall lacking the heteropolysaccharide layer. These cells can be gently disrupted to obtain protoplasts or lysed to yield intact genomic DNA and RNA. Experiments are in progress to develop a gene transfer system in this bacterial species. Methods are being developed and refined for the efficient plating of M. thermophila on defined media, for chemical mutagenesis, and for the isolation of mutants defective in acetate utilization. Chromosomal DNA libraries have been constructed from M. thermophila and are being used to clone genes involved in the acetate utilization pathway (e.g. carbon monoxide dehydrogenase). Once cloned, analysis of the molecular mechanisms responsible for their regulatory control will be performed. These studies should aid our understanding of the pathway for acetate utilization in M. thermophila and serve as a model for elucidating regulatory mechanisms in the acetotrophic methanogens.

  6. Methanoregula boonei gen. nov., sp. nov., an acidiphilic methanogen isolated from an acidic peat bog.

    PubMed

    Bräuer, Suzanna L; Cadillo-Quiroz, Hinsby; Ward, Rebekah J; Yavitt, Joseph B; Zinder, Stephen H

    2011-01-01

    A novel acidiphilic, hydrogenotrophic methanogen, designated strain 6A8(T), was isolated from an acidic (pH 4.0-4.5) and ombrotrophic (rain-fed) bog located near Ithaca, NY, USA. Cultures were dimorphic, containing thin rods (0.2-0.3 μm in diameter and 0.8-3.0 μm long) and irregular cocci (0.2-0.8 μm in diameter). The culture utilized H(2)/CO(2) to produce methane but did not utilize formate, acetate, methanol, ethanol, 2-propanol, butanol or trimethylamine. Optimal growth conditions were near pH 5.1 and 35 °C. The culture grew in basal medium containing as little as 0.43 mM Na(+) and growth was inhibited completely by 50 mM NaCl. To our knowledge, strain 6A8(T) is one of the most acidiphilic (lowest pH optimum) and salt-sensitive methanogens in pure culture. Acetate, coenzyme M, vitamins and yeast extract were required for growth. It is proposed that a new genus and species be established for this organism, Methanoregula boonei gen. nov., sp. nov. The type strain of Methanoregula boonei is 6A8(T) (=DSM 21154(T) =JCM 14090(T)). PMID:20154331

  7. Effects of auxiliary carbon sources and electron acceptors on methanogenic degradation of chlorinated phenols

    SciTech Connect

    Haeggblom, M.M.; Rivera, M.D.; Young, L.Y. )

    1993-08-01

    The authors studied the degradation of chlorinated phenols under methanogenic conditions by establishing enrichment cultures on 4-chlorophenol and 2,4-dichlorophenol with or without a supplementary substrate. p-Cresol was chosen as a nonchlorinated aromatic compound structurally similar to the chlorophenols, and propionate was chosen as a readily utilizable carbon source. 2,4-Dichlorophenol was dechlorinated to 4-chlorophenol, which was degraded without further detection of metabolites. The rates of chlorophenols and supplementary substrates. The addition of p-cresol or propionate as an auxiliary carbon source enhanced the rate of 4-chlorophenol degradation. Methanogenic cultures capable of ortho dechlorination were repeatedly subcultured by dilution into fresh media and refeeding of 2,6-dichlorophenol and either p-cresol or propionate as auxiliary substrates. 2,6-Dichlorophenol was sequentially dechlorinated to 2-chlorophenol and phenol and ultimately mineralized to methane and carbon dioxide. Cultures adapted to 2,4- or 2,6-dichlorophenol also readily dechlorinated other dichlorophenols containing an ortho chlorine. The alternative electron acceptors nitrate, sulfite, and thiosulfate completely inhibited dechlorination of 2,6-dichlorophenol, whereas sulfate slowed the dechlorination rate.

  8. Decolorization of a reactive copper-phthalocyanine dye under methanogenic conditions.

    PubMed

    Beydili, M I; Matthews, R D; Pavlostathis, S G

    2001-01-01

    The objective of this research was to assess the biological decolorization of the copper-phthalocyanine dye Reactive Blue 7 (RB7) under methanogenic conditions using a mixed, methanogenic culture in a repetitive dye addition batch assay. The initial rate of decolorization was 13.2 mg/L-d and 5.7 mg/L-d for the first and second dye addition, respectively. For an initial RB7 concentration of ca. 300 mg/L, the extent of decolorization remained constant (about 62%) for each repetitive RB7 addition and resulted in a residual color build up. Declining absorbance ratio values (A664/A620) with increasing incubation time confirmed that the observed color removal was due to transformation as opposed to adsorption on the biomass. Chemical decolorization assays using sodium dithionite as the reducing agent resulted in similar absorbance spectra to that obtained after biological decolorization. In addition, in both the chemical and biological decolorization assays, partial oxidation of the reduced dye solution upon exposure to air resulted in higher residual color, indicating that the reduction and decolorization of RB7 are partially reversible. These results also suggest that RB7 reduction and decolorization both chemically and biologically most likely followed a similar reduction mechanism. PMID:11380199

  9. Stable carbon isotope fractionation by methanogens growing on different Mars regolith analogs

    NASA Astrophysics Data System (ADS)

    Sinha, Navita; Kral, Timothy A.

    2015-07-01

    In order to characterize stable carbon (13C/12C) isotope fractionation of metabolically produced methane by methanogens in martian settings, Methanothermobacter wolfeii, Methanosarcina barkeri, and Methanobacterium formicicum were cultured on four different Mars regolith analogs - JSC Mars-1, Artificial Mars Simulant, montmorillonite, and Mojave Mars Simulant - and also in their growth supporting media. These chemoautotrophic methanogens utilize CO2 for their carbon source and H2 for their energy source. When compared to the carbon isotope signature of methane when grown on their respective growth media, M. wolfeii and M. barkeri demonstrated variability in carbon isotope fractionation values during methanogenesis on the Mars analogs, while M. formicicum showed subtle or negligible difference in carbon isotope fractionation values. Interestingly, M. wolfeii and M. barkeri have shown relatively consistent enriched values of 12C on montmorillonite, a kind of clay found on Mars, compared to other Mars regolith analogs. In general, M. barkeri showed large carbon isotope fractionation compared to M. wolfeii and M. formicicum during methanognesis on various kinds of analogs. Stable carbon isotope fractionation is one of the techniques used to infer different origins, environments, and pathways of methanogensis. The results obtained in this novel research can provide clues to determine ambiguous sources of methane on Mars.

  10. A novel group of abyssal methanogenic archaebacteria (Methanopyrus) growing at 110 °C

    NASA Astrophysics Data System (ADS)

    Huber, R.; Kurr, M.; Jannasch, H. W.; Stetter, K. O.

    1989-12-01

    THE organisms with the highest growth temperature known so far are members of the archaebacterial genus Pyrodictium1'2. These anaerobic sulphur reducers thrive at temperatures of up to 110 °C within a shallow hydrothermal system off Vulcano, Italy. Only a few hyperthermophilic methanogens are known-members of the genus Methanothermus, which grow exclusively within terrestrial fields of fumaroles from which sulphurous gas is emitted and show an upper growth temperature of 97 °C (ref. 3), and some members of the genus Methanococcus, which grow within deep-sea hydro-thermal systems at temperatures up to about 90 °C (ref. 4). We have now isolated a novel group of methanogenic archaebacteria growing at least at 110°C from sediment samples taken by the research submersible Alvin at the Guaymas Basin hot vents (Gulf of California). This finding demonstrates the unexpected biogenic methanogenesis at temperatures above 100 °C, and, in view of biogeochemistry, could explain isotope discrimination at temperatures that were thought to be unfavourable for biological methanogenesis.

  11. Biodegradation of polychlorinated methanes in methanogenic systems. Annual report, 1 Dec 88-1 Mar 89

    SciTech Connect

    Freedman, D.L.

    1991-01-01

    This research investigated biodegradation of several halogenated methanes under methanogenic conditions. Biodegradation of dichloromethane to C02 and acetic acid (both environmentally acceptable products) was demonstrated in a fixed-film reactor, operated at 20 deg C, a residence time as low as 0.25 day, and an influent concentration of 91 micro M. The biodegradability of chloroform was examined in a dichloromethane-degrading enrichment culture. Sustained consumption of chloroform (approximately 8.5 micron M) was achieved only when vitamin B 12 was also added. Initially, equimolar amounts of chloroform and B 12 were added; when the amount of B 12 added was gradually decreased to zero, chloroform degradation continued at the same rapid rate. Biodegradation of 14C chloroform yielded approximately 77% C02, 10% carbon monoxide, 7% soluble compounds (about 15% of which consisted of acetate), and 2% nonsoluble compounds; no 14CH4 was formed. Brominated methanes (dibromomethane, bromochloromethane, and bromomethane) degraded much more slowly' if at all, than dichloromethane, they inhibited methanogenesis and dichloromethane degradation. Sustained degradation of chloromethane was demonstrated for an extended period without the need for an electron donor. Preliminary results suggest chloromethane serves as an electron donor under methanogenic conditions, just as dichloromethane does. Biotransformation of carbon tetrachloride was also demonstrated in the dichloromethane-degrading enrichment culture.

  12. Squalenes, phytanes and other isoprenoids as major neutral lipids of methanogenic and thermoacidophilic 'archaebacteria'

    NASA Technical Reports Server (NTRS)

    Tornabene, T. G.; Langworthy, T. A.; Holzer, G.; Oro, J.

    1979-01-01

    The neutral lipids from nine species of methanogenic bacteria (five methanobacilli, two methanococci, a methanospirillum and a methanosarcina) and two thermoacidophilic bacteria (Thermo-plasma and Sulfolobus) have been analyzed. The neutral lipids were found to comprise a wide range (C14 to C30) of polyisoprenyl hydrocarbons with varying degrees of saturation. The principal components represented the three major isoprenoid series (C20 phytanyl, C25 pentaisoprenyl, and C30 squalenyl), in contrast with the neutral lipids of extreme halophiles, which consist predominantly of C2O (phytanyl, geranylgeraniol), C30 (squalenes), C40 (carotenes) and C50 (bacterioruberins compounds), as reported by Kates (1978). These results, which indicate strong general similarities between genetically diverse organisms, support the classification of these organisms in a separate phylogenetic group. The occurrence of similar isoprenoid compounds in petroleum and ancient sediments and the fact that the methanogens, halophiles and thermoacidophiles live in conditions presumed to have prevailed in archaen times suggest that the isoprenoid compounds in petroleum compounds and sediment may have been directly synthesized by organisms of this type

  13. Interaction of anammox bacteria and inactive methanogenic granules under high nitrogen selective pressure.

    PubMed

    Ni, Shou-Qing; Fessehaie, Anania; Lee, Po-Heng; Gao, Bao-Yu; Xu, Xing; Sung, Shihwu

    2010-09-01

    Granular anammox reactors usually adopted anaerobic/aerobic granules as source sludge, in which the washout of other species and enrichment of anammox biomass were very slow because of the competition of the coexisting bacteria. In this study, inactive methanogenic granules were proved to be suitable for rapid anammox granulation under high nitrogen concentrations by investigating their interaction with anammox bacteria. The start-up nitrite concentration was significantly higher than the published toxic level for anammox bacteria and other lab-scale studies. The nitrogen loading rate increased from 141 to 480 mg/L/d in 120 days operation with a total nitrogen removal efficiency of 96.0+/-0.6%. Anammox granules with a diameter of 1.3+/-0.4mm were observed over the course of three months. Molecular analysis showed that over 67% of the cells in the anammox granules were anammox bacteria after 90 days. The accommodations and proliferations of anammox bacteria in the inactive methanogenic granules might be the main reason for the high anammox purity in a short period. The important role of the extracellular polymer in the granule structure was observed via morphological observation. PMID:20444597

  14. The carbon isotope biogeochemistry of acetate from a methanogenic marine sediment

    NASA Technical Reports Server (NTRS)

    Blair, N. E.; Carter, W. D., Jr.

    1992-01-01

    The delta C-13 value of porewater acetate isolated from the anoxic sediments of Cape Lookout Bight (North Carolina) ranged from -17.6 percent in the sulfate reduction zone to -2.8 percent in the underlying methanogenic zone. The large C-13 enrichment in the sulfate-depleted sediments appears to be associated with the dissimilation of acetate to CH4 and CO2. Fractionation factors for that process were estimated to be 1.032 +/- 0.014 and 1.036 +/- 0.019 for the methyl and carboxyl groups. A subsurface maximum in delta C-13 of the total acetate molecule, as well as the methyl and carboxyl carbons at 10-15 cm depth within the sediment column, indicate that changes in the relative rates of acetate cycling pathways occur in the methanogenic zone. The methyl group of the acetate was depleted in C-13 by 7-14 percent relative to the carboxyl moiety. The intramolecular heterogeneity may be the result of both synthetic and catabolic isotope effects.

  15. Transformation of indole by methanogenic and sulfate-reducing microorganisms isolated from digested sludge

    SciTech Connect

    Shanker, R.; Bollag, J.M. )

    1990-01-01

    In the present study, mineralization of an aromatic N-heterocyclic molecule, indole, by microorganisms present in anaerobically digested sewage sludge was examined. The first step in indole mineralization was the formation of a hydroxylated intermediate, oxindole. The rate of transformation of indole to oxindole and its subsequent disappearance was dependent on the concentration of inoculum and indole and the incubation temperature. Methanogenesis appeared to be the dominant process in the mineralization of indole in 10% digested sludge even in the presence of high concentrations of sulfate. Enrichment of the digested sludge with sulfate as an electron acceptor allowed the isolation of a metabolically stable mixed culture of anaerobic bacteria which transformed indole to oxindole and acetate, and ultimately to methane and carbon dioxide. This mixed culture exhibited a predominance of sulfate-reducers over methanogens with more than 75% of the substrate mineralized to carbon dioxide. The investigation demonstrates that indole can be transformed by both methanogenic and sulfate-reducing microbial populations.

  16. Effects of biopretreatment on pyrolysis behaviors of corn stalk by methanogen.

    PubMed

    Wang, Tipeng; Ye, Xiaoning; Yin, Jun; Lu, Qiang; Zheng, Zongming; Dong, Changqing

    2014-07-01

    The study investigated the effects of methanogen pretreatment on pyrolysis behaviors of corn stalk (CS) by using Py-GC/MS analysis and thermogravimetric analysis. Results indicated that biopretreatment changed considerably the pyrolysis behaviors of CS from four weight loss stages to two weight loss stages. Increasing biopretreatment time from 5 days to 25 days enhanced the kinds and contents of chemicals in volatile products. In pyrolysis products, the contents of sugars, linear ketones and furans decreased from 1.43%, 12.60% and 7.38% to 1.25%, 10.22% and 3.25%, respectively, and the contents of phenols increased from 15.08% to 27.84%. The most content change from 6.83% to 13.63% indicated that methanogen pretreatment improved the pyrolysis selectivity of CS to product the 4-VP, but it was disadvantageous to 5-hydroxymethyl furfural, levoglucose and furfural. The changes of chemical compositions and structure of CS after biopretreatment were the main reason of the differences. PMID:24855936

  17. The complete genome sequence of the rumen methanogen Methanobacterium formicicum BRM9

    PubMed Central

    2014-01-01

    Methanobacterium formicicum BRM9 was isolated from the rumen of a New Zealand Friesan cow grazing a ryegrass/clover pasture, and its genome has been sequenced to provide information on the phylogenetic diversity of rumen methanogens with a view to developing technologies for methane mitigation. The 2.45 Mb BRM9 chromosome has an average G + C content of 41%, and encodes 2,352 protein-coding genes. The genes involved in methanogenesis are comparable to those found in other members of the Methanobacteriaceae with the exception that there is no [Fe]-hydrogenase dehydrogenase (Hmd) which links the methenyl-H4MPT reduction directly with the oxidation of H2. Compared to the rumen Methanobrevibacter strains, BRM9 has a much larger complement of genes involved in determining oxidative stress response, signal transduction and nitrogen fixation. BRM9 also has genes for the biosynthesis of the compatible solute ectoine that has not been reported to be produced by methanogens. The BRM9 genome has a prophage and two CRISPR repeat regions. Comparison to the genomes of other Methanobacterium strains shows a core genome of ~1,350 coding sequences and 190 strain-specific genes in BRM9, most of which are hypothetical proteins or prophage related. PMID:25780506

  18. Toxicity of N-substituted aromatics to acetoclastic methanogenic activity in granular sludge.

    PubMed Central

    Donlon, B A; Razo-Flores, E; Field, J A; Lettinga, G

    1995-01-01

    N-substituted aromatics are important priority pollutants entering the environment primarily through anthropogenic activities associated with the industrial production of dyes, explosives, pesticides, and pharmaceuticals. Anaerobic treatment of wastewaters discharged by these industries could potentially be problematical as a result of the high toxicity of N-substituted aromatics. The objective of this study was to examine the structure-toxicity relationships of N-substituted aromatic compounds to acetoclastic methanogenic bacteria. The toxicity was assayed in serum flasks by measuring methane production in granular sludge. Unacclimated cultures were used to minimize the biotransformation of the toxic organic chemicals during the test. The nature and the degree of the aromatic substitution were observed to have a profound effect on the toxicity of the test compound. Nitroaromatic compounds were, on the average, over 500-fold more toxic than their corresponding aromatic amines. Considering the facile reduction of nitro groups by anaerobic microorganisms, a dramatic detoxification of nitroaromatics towards methanogens can be expected to occur during anaerobic wastewater treatment. While the toxicity exerted by the N-substituted aromatic compounds was closely correlated with compound apolarity (log P), it was observed that at any given log P, N-substituted phenols had a toxicity that was 2 orders of magnitude higher than that of chlorophenols and alkylphenols. This indicates that toxicity due to the chemical reactivity of nitroaromatics is much more important than partitioning effects in bacterial membranes. PMID:8526501

  19. Investigations of methane emissions from rice cultivation in Indian context.

    PubMed

    Anand, Shalini; Dahiya, R P; Talyan, Vikash; Vrat, Prem

    2005-05-01

    The increasing demand of the growing population requires enhancement in the production of rice. This has a direct bearing on the global environment since the rice cultivation is one of the major contributors to the methane emissions. As the rice cultivation is intensified with the current practices and technologies, the methane fluxes from paddy fields will substantially rise. Improved high yielding rice varieties together with efficient cultivation techniques will certainly contribute to the curtailment of the methane emission fluxes. In this paper, the system dynamic approach is used for estimating the methane emissions from rice fields in India till the year 2020. Mitigation options studied for curtailing the methane emissions include rice production management, use of low methane emitting varieties of rice, water management and fertilizer amendment. The model is validated quantitatively and sensitivity tests are carried out to examine the robustness of the model. PMID:15788188

  20. Stable acetate production in extreme-thermophilic (70°C) mixed culture fermentation by selective enrichment of hydrogenotrophic methanogens

    PubMed Central

    Zhang, Fang; Zhang, Yan; Ding, Jing; Dai, Kun; van Loosdrecht, Mark C. M.; Zeng, Raymond J.

    2014-01-01

    The control of metabolite production is difficult in mixed culture fermentation. This is particularly related to hydrogen inhibition. In this work, hydrogenotrophic methanogens were selectively enriched to reduce the hydrogen partial pressure and to realize efficient acetate production in extreme-thermophilic (70°C) mixed culture fermentation. The continuous stirred tank reactor (CSTR) was stable operated during 100 days, in which acetate accounted for more than 90% of metabolites in liquid solutions. The yields of acetate, methane and biomass in CSTR were 1.5 ± 0.06, 1.0 ± 0.13 and 0.4 ± 0.05 mol/mol glucose, respectively, close to the theoretical expected values. The CSTR effluent was stable and no further conversion occurred when incubated for 14 days in a batch reactor. In fed-batch experiments, acetate could be produced up to 34.4 g/L, significantly higher than observed in common hydrogen producing fermentations. Acetate also accounted for more than 90% of soluble products formed in these fed-batch fermentations. The microbial community analysis revealed hydrogenotrophic methanogens (mainly Methanothermobacter thermautotrophicus and Methanobacterium thermoaggregans) as 98% of Archaea, confirming that high temperature will select hydrogenotrophic methanogens over aceticlastic methanogens effectively. This work demonstrated a potential application to effectively produce acetate as a value chemical and methane as an energy gas together via mixed culture fermentation. PMID:24920064

  1. Stable acetate production in extreme-thermophilic (70°C) mixed culture fermentation by selective enrichment of hydrogenotrophic methanogens

    NASA Astrophysics Data System (ADS)

    Zhang, Fang; Zhang, Yan; Ding, Jing; Dai, Kun; van Loosdrecht, Mark C. M.; Zeng, Raymond J.

    2014-06-01

    The control of metabolite production is difficult in mixed culture fermentation. This is particularly related to hydrogen inhibition. In this work, hydrogenotrophic methanogens were selectively enriched to reduce the hydrogen partial pressure and to realize efficient acetate production in extreme-thermophilic (70°C) mixed culture fermentation. The continuous stirred tank reactor (CSTR) was stable operated during 100 days, in which acetate accounted for more than 90% of metabolites in liquid solutions. The yields of acetate, methane and biomass in CSTR were 1.5 +/- 0.06, 1.0 +/- 0.13 and 0.4 +/- 0.05 mol/mol glucose, respectively, close to the theoretical expected values. The CSTR effluent was stable and no further conversion occurred when incubated for 14 days in a batch reactor. In fed-batch experiments, acetate could be produced up to 34.4 g/L, significantly higher than observed in common hydrogen producing fermentations. Acetate also accounted for more than 90% of soluble products formed in these fed-batch fermentations. The microbial community analysis revealed hydrogenotrophic methanogens (mainly Methanothermobacter thermautotrophicus and Methanobacterium thermoaggregans) as 98% of Archaea, confirming that high temperature will select hydrogenotrophic methanogens over aceticlastic methanogens effectively. This work demonstrated a potential application to effectively produce acetate as a value chemical and methane as an energy gas together via mixed culture fermentation.

  2. DEPENDENCE OF A HIGH-RATE, PCE-DECHLORINATING ENRICHMENT CULTURE ON METHANOGENIC ACTIVITY. (R825549C053)

    EPA Science Inventory

    The role served by the presence of methanogenic activity within a tetrachloroethene (PCE)-dechlorinating culture was investigated through a series of supplementation experiments. An acclimated lactate-enrichment culture (LEC 1) capable of rapidly converting PCE to ethene was s...

  3. MICROBIAL REDUCTIVE DECHLORINATION OF HEXACHLORO-1,3-BUTADIENE IN A METHANOGENIC ENRICHMENT CULTURE. (R825513C007)

    EPA Science Inventory

    Sequential reductive dechlorination of hexachloro-1,3-butadiene (HCBD) was achieved by a mixed, methanogenic culture enriched from a contaminated estuarine sediment. Both methanol and lactate served as carbon and electron sources. Methanol was stoichiometrically converted to m...

  4. Thioredoxin-linked redox control of metabolism in Methanocaldococcus jannaschii, an evolutionarily deeply-rooted hyperthermophilic methanogenic archaeon

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Thioredoxin (Trx), a small redox protein, controls multiple processes in eukaryotes and bacteria by changing the thiol redox status of selected proteins. We have investigated this aspect in methanarchaea. These ancient methanogens produce methane almost exclusively from H2 plus CO2 carried approxima...

  5. [Impacts of cultivated land conversion on cultivated land productivity in China: prediction and analysis].

    PubMed

    Jiang, Qun-ou; Deng, Xiang-zheng; Lin, Ying-zhi; Cui, Yong-wei

    2010-12-01

    This paper simulated the spatial patterns of cultivated land in China under the future scenario by using the Dynamics of Land System (DLS) model, and then estimated the cultivated land productivity at the grid pixel dimensions based on the Estimation System of Land Production (ESLP). In addition, the spatial patterns of cultivated land productivity in each of China agro-ecological zones were analyzed. On this basis, this paper predicted the impacts of cultivated land conversion on the cultivated land production in China in 2000-2020, and identified the major affecting factors on the cultivated land production. The research results indicated that the impact of improving the cultivated land productivity on the cultivated land production would be wunch more remarkable than that from the magnitude of cultivated land conversion in regions where there were high potential to imrprove the cultivated land productivity. However, in the regions with nearly no room to improve the productivity, cultivated land conversion would produce more apparent impacts on the total cultivated land production. In this sense, it was of significance for the national food security in China to adjust the cultivated land conversion to ensure the 0.12 billion hm2 of cultivated land, and to increase investment and improve management level to increase per unit grain yield. PMID:21442997

  6. Bacteria and Methanogens Differ along the Gastrointestinal Tract of Chinese Roe Deer (Capreolus pygargus)

    PubMed Central

    Li, Zhipeng; Zhang, Zhigang; Xu, Chao; Zhao, Jingbo; Liu, Hanlu; Fan, Zhongyuan; Yang, Fuhe; Wright, André-Denis G.; Li, Guangyu

    2014-01-01

    The current study provides the insight into the bacteria in the gastrointestinal tract (GIT) and methanogens presented in the rumen and cecum of the Chinese roe deer (Capreolus pygargus). The ruminal, ileal, cecal, and colonic contents, as well as feces, were obtained from each of the three, free-range, roe deer ingesting natural pasture after euthanasia. For the bacterial community, a total of 697,031 high-quality 16S rRNA gene sequences were generated using high-throughput sequencing, and assigned to 2,223 core operational taxonomic units (OTUs) (12 bacterial phyla and 87 genera). The phyla Firmicutes (51.2%) and Bacteroidetes (39.4%) were the dominant bacteria in the GIT of roe deer. However, the bacterial community in the rumen was significantly (P<0.01) different from the other sampled regions along the GIT. Secondly, Prevotella spp., Anaerovibrio spp., and unidentified bacteria within the families Veillonellaceae and Paraprevotellaceae were more abundant in the rumen than in the other regions. Unidentified bacteria within the family Enterobacteriaceae, Succinivibrio spp., and Desulfovibrio spp. were more predominant in the colon than in other regions. Unidentified bacteria within the family Ruminococcaceae, and Bacteroides spp. were more prevalent in the ileum, cecum and fecal pellets. For methanogens in the rumen and cecum, a total of 375,647 high quality 16S rRNA gene sequences were obtained and assigned to 113 core OTUs. Methanobrevibacter millerae was the dominant species accounting for 77.3±7.4 (S.E) % and 68.9±4.4 (S.E) % of total sequences in the rumen and cecum of roe deer, respectively. However, the abundance of Methanobrevibacter smithii was higher in the rumen than in the cecum (P = 0.004). These results revealed that there was intra variation in the bacterial community composition across the GIT of roe deer, and also showed that the methanogen community in the rumen differed from that in the cecum. PMID:25490208

  7. Seaweed cultivation: Traditional way and its reformation

    NASA Astrophysics Data System (ADS)

    Fei, Xiu-Geng; Bao, Ying; Lu, Shan

    1999-09-01

    Seaweed cultivation or phycoculture has been developed rather fast in recent years. The total production of cultivated seaweed at present is about 6250×103 tons fresh weight. The total cultivation area is estimated as 200×103 hectare. The annual total value of cultivated seaweeds has been estimated to be more than 3 billion US dollars. Phycoculture provides many job opportunities for the coastal region people, has the potential to improve marine environments and thus even induce global change. All traditional cultivation methods and techniques are based on or start from the individual plant or the cultivated seaweed population. Modern biological science and biotechnology achievements have benefited agriculture a lot, but traditional seaweed cultivation has not changed much since its founding. This is because seaweed cultivation has been quite conservative for quite a long period and has accumulated many problems requiring solution. Four main problems might be the most universal ones holding back further development of the industry. New ways of seaweed cultivation must be developed, new techniques must be perfected, and new problems solved. This paper mainly discusses the main problems of traditional seaweed cultivation at present and its possible further development and reformation in the future.

  8. Formation of unilamellar liposomes from total polar lipid extracts of methanogens.

    PubMed Central

    Choquet, C G; Patel, G B; Beveridge, T J; Sprott, G D

    1992-01-01

    Unilamellar liposomes were formed by controlled detergent dialysis of mixed micelles consisting of acetone-insoluble total polar lipids extracted from various methanogens and the detergent n-octyl-beta-D-glucopyranoside. The final liposome populations were studied by dynamic light scattering and electron microscopy. Unilamellar liposomes with mean diameters smaller than 100 nm were obtained with lipid extracts of Methanococcus voltae, Methanosarcina mazei, Methanosaeta concilii, and Methanococcus jannaschii (grown at 50 degrees C), whereas larger (greater than 100-nm) unilamellar liposomes were obtained with lipid extracts of M. jannaschii grown at 65 degrees C. These liposomes were shown to be closed intact vesicles capable of retaining entrapped [14C]sucrose for extended periods of time. With the exception of Methanospirillum hungatei liposomes, all size distributions of the different liposome populations were fairly homogeneous. Images PMID:1444403

  9. [Methanogenic destruction of (amino)aromatic compounds by anaerobic microbial communities].

    PubMed

    Lin'kova, Iu V; D'iakonova, A T; Gladchenko, M A; Kaliuzhnyĭ, S A; Kotova, I B; Stams, A; Netrusov, A I

    2011-01-01

    Destruction of a number of aromatic substrates by anaerobic microbial communities was studied. Active methanogenic microbial communities decomposing aminoaromatic acids and azo dyes into CH4 and CO2 were isolated. Products of primary conversion were found to be 2-hydroxybenzyl and benzyl alcohols gradually transforming into benzoate. It was shown that isolated microbial communities are capable of converting the initial substrates--benzyl alcohol, benzoate, salicylic acid, and golden yellow azo dye--into biogas without a lag-phase but with different velocities. Aromatic and linear intermediates of biodestruction of aromatic amines by obtained enrichment cultures were determined for the first time. Selective effect of aromatic substrates on a microbial community that was expressed in decrease in diversity and gradual change of dominant morphotypes was revealed. PMID:22232897

  10. Methanomassiliicoccus luminyensis gen. nov., sp. nov., a methanogenic archaeon isolated from human faeces.

    PubMed

    Dridi, Bédis; Fardeau, Marie-Laure; Ollivier, Bernard; Raoult, Didier; Drancourt, Michel

    2012-08-01

    During attempts to obtain novel, human-associated species of the domain Archaea, a coccoid micro-organism, designated strain B10(T), was isolated in pure culture from a sample of human faeces collected in Marseille, France. On the basis of its phenotypic characteristics and 16S rRNA and mcrA gene sequences, the novel strain was classified as a methanogenic archaeon. Cells of the strain were non-motile, Gram-staining-positive cocci that were approximately 850 nm in diameter and showed autofluorescence at 420 nm. Cells were lysed by 0.1% (w/v) SDS. With hydrogen as the electron donor, strain B10(T) produced methane by reducing methanol. The novel strain was unable to produce methane when hydrogen or methanol was the sole energy source. In an atmosphere containing CO(2), strain B10(T) could not produce methane from formate, acetate, trimethylamine, 2-butanol, 2-propanol, cyclopentanol, 2-pentanol, ethanol, 1-propanol or 2,3-butanediol. Strain B10(T) grew optimally with 0.5-1.0% (w/v) NaCl, at pH 7.6 and at 37 °C. It required tungstate-selenite for growth. The complete genome of the novel strain was sequenced; the size of the genome was estimated to be 2.05 Mb and the genomic DNA G+C content was 59.93 mol%. In phylogenetic analyses based on 16S rRNA gene sequences, the highest sequence similarities (98.0-98.7%) were seen between strain B10(T) and several uncultured, methanogenic Archaea that had been collected from the digestive tracts of a cockroach, a chicken and mammals. In the same analysis, the non-methanogenic 'Candidatus Aciduliprofundum boonei' DSM 19572 was identified as the cultured micro-organism that was most closely related to strain B10(T) (83.0% 16S rRNA gene sequence similarity). Each of the three treeing algorithms used in the analysis of 16S rRNA gene sequences indicated that strain B10(T) belongs to a novel order that is distinct from the Thermoplasmatales. The novel strain also appeared to be distinct from Methanosphaera stadtmanae DSM 3091(T) (72

  11. Methanogenic population dynamics during start-up of anaerobic digesters treating municipal solid waste and biosolids

    SciTech Connect

    Griffin, M.E.; McMahon, K.D.; Mackie, R.I.; Raskin, L.

    1998-02-05

    An aggressive start-up strategy was used to initiate codigestion in two anaerobic, continuously mixed bench-top reactors at mesophilic (37 C) and thermophilic (55 C) conditions. The digesters were inoculated with mesophilic anaerobic sewage sludge and cattle manure and were fed a mixture of simulated municipal solid waste and biosolids in proportions that reflect US production rates. The design organic loading rate was 3.1 kg volatile solids/m{sup 3}/day and the retention time was 20 days. Ribosomal RNA-targeted oligonucleotide probes were used to determine the methanogenic community structure in the inocula and the digesters. Chemical analyses were performed to evaluate digester performance. The aggressive start-up strategy was successful for the thermophilic reactor, despite the use of a mesophilic inoculum.

  12. Methanogenic Activity and Structural Characteristics of the Microbial Biofilm On a Needle-Punched Polyester Support

    PubMed Central

    Harvey, Martin; Forsberg, Cecil W.; Beveridge, Terry J.; Pos, Jack; Ogilvie, John R.

    1984-01-01

    In a downflow stationary fixed-film anaerobic reactor receiving a swine waste influent, few bacteria were observed to be tightly adherent to the surfaces of the needle-punched polyester support material. However, there was a morphologically complex, dense population of bacteria trapped within the matrix. Frequently large microcolonies of a uniform morphological type of bacteria were observed. These were particularly evident for methanosarcina-like bacteria which grew forming large aggregates of unseparated cells. Leafy deposits of electron-dense, calcium- and phosphorus-enriched material coated the polyester matrix and some cells. As the biofilm matured there was more extensive mineral deposition which completely entrapped cells. The entrapped cells appeared to autolyze, and many were partially degraded. Further impregnation of the matrix with minerals and apparent cell death may eventually have a deleterious effect on the methanogenic activity of the biofilm. Images PMID:16346629

  13. Isolation and Characterization of a Halophilic Methanogen from Great Salt Lake †

    PubMed Central

    Paterek, J. Robert; Smith, Paul H.

    1985-01-01

    A halophilic methanogenic microorganism isolated from sediments collected from the southern arm of Great Salt Lake, Utah, is described. Cells were irregular, nonmotile cocci approximately 1.0 μm in diameter and stained gram negative. Colonies from anaerobic plates and roll tubes were foamy, circular, and cream-yellow. Methanol, methylamine, dimethylamine, and trimethylamine supported growth and methanogenesis. Hydrogen-carbon dioxide, formate, and acetate were not utilized. Sodium and magnesium were required for growth; the optimum NaCl concentration ranged between 1.0 and 2.0 M, with the minimum doubling time occurring at 2.0 M. The optimum growth temperature was 35°C, with maximum growth rate occurring at pH 7.5. The DNA base composition was 48.5 mol% guanine + cytosine. SLP is the type strain designation (= ATCC 35705). Images PMID:16346919

  14. Chemical and carbon isotopic composition of dissolved organic carbon in a regional confined methanogenic aquifer

    USGS Publications Warehouse

    Aravena, R.; Wassenaar, L.I.; Spiker, E. C.

    2004-01-01

    This study demonstrates the advantage of a combined use of chemical and isotopic tools to understand the dissolved organic carbon (DOC) cycle in a regional confined methanogenic aquifer. DOC concentration and carbon isotopic data demonstrate that the soil zone is a primary carbon source of groundwater DOC in areas close to recharge zones. An in-situ DOC source linked to organic rich sediments present in the aquifer matrix is controlling the DOC pool in the central part of the groundwater flow system. DOC fractions, 13C-NMR on fulvic acids and 14C data on DOC and CH4 support the hypothesis that the in-situ DOC source is a terrestrial organic matter and discard the Ordovician bedrock as a source of DOC. ?? 2004 Taylor and Francis Ltd.

  15. Methane formation from fructose by syntrophic associations of Acetobacterium woodii and different strains of methanogens.

    PubMed

    Winter, J U; Wolfe, R S

    1980-01-01

    When Acetobacterium woodii was co-cultured in continuous or in stationary culture with Methanobacterium strain AZ, fructose instead of being converted to 3 mol of acetate was converted to 2 mol of acetate and 1 mol each of carbon dioxide and methane, showing that interspecies hydrogen transfer occurred. In continuous culture the organisms formed a close physical association in clumps; the doubling time for each organism was 6 h at 33 degrees C. Methane mainly was derived from carbon positions 3 and 4 of the sugar, but other carbons also yielded methane; this was shown to be due to carbon dioxide-acetate exchange reactions by A. woodii in a manner similar to that carried out by Clostridium thermoaceticum. Four other methanogens, Methanobacterium M.o.H. and M.o.H. G, Methanobacterium formicicum, and Methanosarcina barkeri (not acetate-adapted) also produced similar results, when co-cultured with A. woodii. PMID:6769417

  16. Isolation and characterization of a halophilic methanogen from Great Salt Lake

    SciTech Connect

    Paterek, J.R.; Smith, P.H.

    1985-10-01

    A halophilic methanogenic microorganism isolated from sediments collected from the southern arm of Great Salt Lake, Utah, is described. Cells were irregular, nonmotile cocci approximately 1.0 ..mu..m in diameter and stained gram negative. Colonies from anaerobic plates and roll tubes were foamy, circular, and cream-yellow. Methanol, methylamine, dimethylamine, and trimethylamine supported growth and methanogenesis. Hydrogen-carbon dioxide, formate, and acetate were not utilized. Sodium and magnesium were required for growth; the optimum NaCl concentration ranged between 1.0 and 2.0 M, with the minimum doubling time occurring at 2.0 M. The optimum growth temperature was 35/sup 0/C, with maximum growth occurring at pH 7.5. The DNA base composition was 48.5 mol% guanine + cytosine. SLP is the type strain designation (= ATCC 35705).

  17. [Cobalt(III)-EDTA]- Reduction by Thermophilic Methanogen Methanothermobacter Thermautotrophicus

    SciTech Connect

    Singh, Rajesh; Dong, Hailiang; Liu, Deng; Marts, Amy R.; Tierney, David L.; Almquist, Catherine B.

    2015-06-30

    Cobalt is a metal contaminant at high temperature radioactive waste disposal sites. In previous studies have largely focused on mesophilic microorganisms to remediate cobalt, despite the presence of thermophilic microorganisms at such sites. In this study,Methanothermobacter thermautotrophicus, a thermophilic methanogen, was used to reduce Co(III) in the form of [Co(III)–EDTA]-. Bioreduction experiments were conducted in a growth medium with H2/CO2 as a growth substrate at initial Co(III) concentrations of 1, 2, 4, 7, and 10 mM. At low Co(III) concentrations (< 4 mM), a complete reduction was observed within a week. Wet chemistry, X-ray absorption near-edge structure (XANES) and electron paramagnetic resonance (EPR) analyses were all consistent in revealing the reduction kinetics. But, at higher concentrations (7 and 10 mM) the reduction extents only reached 69.8% and 48.5%, respectively, likely due to the toxic effect of Co(III) to the methanogen cells as evidenced by a decrease in total cellular protein at these Co(III) concentrations. Methanogenesis was inhibited by Co(III) bioreduction, possibly due to impaired cell growth and electron diversion from CO2 to Co(III). Overall, our results demonstrated the ability of M. thermautotrophicus to reduce Co(III) to Co(II) and its potential application for remediating 60Co contaminant at high temperature subsurface radioactive waste disposal sites.

  18. Response of a Rice Paddy Soil Methanogen to Syntrophic Growth as Revealed by Transcriptional Analyses

    PubMed Central

    Liu, Pengfei; Yang, Yanxiang; Lü, Zhe

    2014-01-01

    Members of Methanocellales are widespread in paddy field soils and play the key role in methane production. These methanogens feature largely in these organisms' adaptation to low H2 and syntrophic growth with anaerobic fatty acid oxidizers. The adaptive mechanisms, however, remain unknown. In the present study, we determined the transcripts of 21 genes involved in the key steps of methanogenesis and acetate assimilation of Methanocella conradii HZ254, a strain recently isolated from paddy field soil. M. conradii was grown in monoculture and syntrophically with Pelotomaculum thermopropionicum (a propionate syntroph) or Syntrophothermus lipocalidus (a butyrate syntroph). Comparison of the relative transcript abundances showed that three hydrogenase-encoding genes and all methanogenesis-related genes tested were upregulated in cocultures relative to monoculture. The genes encoding formylmethanofuran dehydrogenase (Fwd), heterodisulfide reductase (Hdr), and the membrane-bound energy-converting hydrogenase (Ech) were the most upregulated among the evaluated genes. The expression of the formate dehydrogenase (Fdh)-encoding gene also was significantly upregulated. In contrast, an acetate assimilation gene was downregulated in cocultures. The genes coding for Fwd, Hdr, and the D subunit of F420-nonreducing hydrogenase (Mvh) form a large predicted transcription unit; therefore, the Mvh/Hdr/Fwd complex, capable of mediating the electron bifurcation and connecting the first and last steps of methanogenesis, was predicted to be formed in M. conradii. We propose that Methanocella methanogens cope with low H2 and syntrophic growth by (i) stabilizing the Mvh/Hdr/Fwd complex and (ii) activating formate-dependent methanogenesis. PMID:24837392

  19. Methanoculleus sediminis sp. nov., a methanogen from sediments near a submarine mud volcano.

    PubMed

    Chen, Sheng-Chung; Chen, Mei-Fei; Lai, Mei-Chin; Weng, Chieh-Yin; Wu, Sue-Yao; Lin, Saulwood; Yang, Tsanyao F; Chen, Po-Chun

    2015-07-01

    A mesophilic, hydrogenotrophic methanogen, strain S3Fa(T), was isolated from sediments collected by Ocean Researcher I cruise ORI-934 in 2010 near the submarine mud volcano MV4 located at the upper slope of south-west Taiwan. The methanogenic substrates utilized by strain S3Fa(T) were formate and H2/CO2 but not acetate, secondary alcohols, methylamines, methanol or ethanol. Cells of strain S3Fa(T) were non-motile, irregular cocci, 0.5-1.0 μm in diameter. The surface-layer protein showed an Mr of 128,000.The optimum growth conditions were 37 °C, pH 7.1 and 0.17 M NaCl. The DNA G+C content of the genome of strain S3Fa(T) was 62.3 mol%. Phylogenetic analysis revealed that strain S3Fa(T) was most closely related to Methanoculleus marisnigri JR1(T) (99.3% 16S rRNA gene sequence similarity). Genome relatedness between strain S3Fa(T) and Methanoculleus marisnigri JR1(T) was computed using both genome-to-genome distance analysis (GGDA) and average nucleotide identity (ANI) with values of 46.3-55.5% and 93.08%, respectively. Based on morphological, phenotypic, phylogenetic and genomic relatedness data, it is evident that strain S3Fa(T) represents a novel species of the genus Methanoculleus, for which the name Methanoculleus sediminis sp. nov. is proposed. The type strain is S3Fa(T) ( = BCRC AR10044(T) = DSM 29354(T)). PMID:25855623

  20. Assessing the impact of alumina nanoparticles in an anaerobic consortium: methanogenic and humus reducing activity.

    PubMed

    Alvarez, Luis H; Cervantes, Francisco J

    2012-09-01

    The impact of γ-Al(2)O(3) nanoparticles (NP) on specific methanogenic activity (SMA) and humus reducing activity (HRA) in an anaerobic consortium was evaluated. SMA in sludge incubations without γ-Al(2)O(3) was always higher compared with those performed in the presence of 100 g/L of γ-Al(2)O(3). Nevertheless, the SMA in incubations with γ-Al(2)O(3) was not completely inhibited, indicating that some methanogenic microorganisms were physiologically active even in the presence of γ-Al(2)O(3) NP during the incubation period (~400 h). SMA and HRA of the anaerobic consortium were also conducted in the presence of γ-Al(2)O(3) NP coated with humic acids (HA). Microbial HA reduction occurred 3.7-fold faster using HA immobilized on γ-Al(2)O(3) NP (HA(Imm)), compared with the control with suspended HA (HA(Sus)). Furthermore, immobilized HA decreased the toxicological effects of γ-Al(2)O(3) NP on methanogenesis. Scanning electron microscopy (SEM) images revealed cell membrane damage in those sludge incubations exposed to uncoated γ-Al(2)O(3) NP. In contrast, cell damage was not observed in incubations with HA-coated γ-Al(2)O(3) NP. Methanogenesis out-competed microbial humus reduction regardless if HA was HA(Imm) or HA(Sus). The present study provides a clear demonstration that HA immobilized in γ-Al(2)O(3) NP are effective terminal electron acceptor for microbial respiration and suggests that HA could mitigate the toxicological effects of metal oxide NP on anaerobic microorganisms. PMID:22159738

  1. Molecular Biomarker-Based Biokinetic Modeling of a PCE-Dechlorinating and Methanogenic Mixed Culture

    SciTech Connect

    Heavner, Gretchen L. W.; Rowe, Annette R.; Mansfeldt, Cresten B.; Pan, Ju Khuan; Gossett, James M.; Richardson, Ruth E.

    2013-04-16

    Bioremediation of chlorinated ethenes via anaerobic reductive dechlorination relies upon the activity of specific microbial population-most notably Dehalococcoides (DHC) strains. In the lab and field Dehalococcoides grow most robustly in mixed communities which usually contain both fermenters and methanogens. Recently, researchers have been developing quantitative molecular biomarkers to aid in field site diagnostics and it is hoped that these biomarkers could aid in the modeling of anaerobic reductive dechlorination. A comprehensive biokinetic model of a community containing Dehalococcoides mccartyi (formerly D. ethenogenes) was updated to describe continuously fed reactors with specific biomass levels based on quantitative PCR (qPCR)-based population data (DNA and RNA). The model was calibrated and validated with subsets of chemical and molecular biological data from various continuous feed experiments (n = 24) with different loading rates of the electron acceptor (1.5 to 482 μeeq/L-h), types of electron acceptor (PCE, TCE, cis-DCE) and electron donor to electron acceptor ratios. The resulting model predicted the sum of dechlorination products vinyl chloride (VC) and ethene (ETH) well. However, VC alone was under-predicted and ETH was over predicted. Consequently, competitive inhibition among chlorinated ethenes was examined and then added to the model. Additionally, as 16S rRNA gene copy numbers did not provide accurate model fits in all cases, we examined whether an improved fit could be obtained if mRNA levels for key functional enzymes could be used to infer respiration rates. The resulting empirically derived mRNA “adjustment factors” were added to the model for both DHC and the main methanogen in the culture (a Methanosaeta species) to provide a more nuanced prediction of activity. Results of this study suggest that at higher feeding rates competitive inhibition is important and mRNA provides a more accurate indicator of a population’s instantaneous

  2. Chloroform degradation in methanogenic methanol enrichment cultures and by Methanosarcina barkeri 227.

    PubMed Central

    Bagley, D M; Gossett, J M

    1995-01-01

    The effects of methanol addition and consumption on chloroform degradation rate and product distribution in methanogenic methanol enrichment cultures and in cultures of Methanosarcina barkeri 227 were investigated. Degradation of chloroform with initial concentrations up to 27.3 microM in enrichment cultures and 4.8 microM in pure cultures was stimulated by the addition of methanol. However, methanol consumption was inhibited by as little as 2.5 microM chloroform in enrichment cultures and 0.8 microM chloroform in pure cultures, suggesting that the presence of methanol, not its exact concentration or consumption rate, was the most significant variable affecting chloroform degradation rate. Methanol addition also significantly increased the number of moles of dichloromethane produced per mole of chloroform consumed. In enrichment cultures, the number of moles of dichloromethane produced per mole of chloroform consumed ranged from 0.7 (methanol consumption essentially uninhibited) to 0.35 (methanol consumption significantly inhibited) to less than 0.2 (methanol not added to the culture). In pure cultures, the number of moles of dichloromethane produced per mole of chloroform consumed was 0.47 when methanol was added and 0.24 when no methanol was added. Studies with [14C]chloroform in both enrichment and pure cultures confirmed that methanol metabolism stimulated dichloromethane production compared with CO2 production. The results indicate that while the addition of methanol significantly stimulated chloroform degradation in both methanogenic methanol enrichment cultures and cultures of M. barkeri 227, the prospects for use of methanol as a growth substrate for anaerobic chloroform-degrading systems may be limited unless the increased production of undesirable chloroform degradation products and the inhibition of methanol consumption can be mitigated. PMID:7574627

  3. Molecular biomarker-based biokinetic modeling of a PCE-dechlorinating and methanogenic mixed culture.

    PubMed

    Heavner, Gretchen L W; Rowe, Annette R; Mansfeldt, Cresten B; Pan, Ju Khuan; Gossett, James M; Richardson, Ruth E

    2013-04-16

    Bioremediation of chlorinated ethenes via anaerobic reductive dechlorination relies upon the activity of specific microbial populations--most notably Dehalococcoides (DHC) strains. In the lab and field Dehalococcoides grow most robustly in mixed communities which usually contain both fermenters and methanogens. Recently, researchers have been developing quantitative molecular biomarkers to aid in field site diagnostics and it is hoped that these biomarkers could aid in the modeling of anaerobic reductive dechlorination. A comprehensive biokinetic model of a community containing Dehalococcoides mccartyi (formerly D. ethenogenes) was updated to describe continuously fed reactors with specific biomass levels based on quantitative PCR (qPCR)-based population data (DNA and RNA). The model was calibrated and validated with subsets of chemical and molecular biological data from various continuous feed experiments (n = 24) with different loading rates of the electron acceptor (1.5 to 482 μeeq/L-h), types of electron acceptor (PCE, TCE, cis-DCE) and electron donor to electron acceptor ratios. The resulting model predicted the sum of dechlorination products vinyl chloride (VC) and ethene (ETH) well. However, VC alone was under-predicted and ETH was over predicted. Consequently, competitive inhibition among chlorinated ethenes was examined and then added to the model. Additionally, as 16S rRNA gene copy numbers did not provide accurate model fits in all cases, we examined whether an improved fit could be obtained if mRNA levels for key functional enzymes could be used to infer respiration rates. The resulting empirically derived mRNA "adjustment factors" were added to the model for both DHC and the main methanogen in the culture (a Methanosaeta species) to provide a more nuanced prediction of activity. Results of this study suggest that at higher feeding rates competitive inhibition is important and mRNA provides a more accurate indicator of a population's instantaneous

  4. Low pressure and desiccation effects on methanogens: Implications for life on Mars

    NASA Astrophysics Data System (ADS)

    Kral, Timothy A.; Altheide, Travis S.; Lueders, Adrienne E.; Schuerger, Andrew C.

    2011-02-01

    Conditions on the surface of Mars would appear to be too hostile for life as we know it. But the subsurface is another matter. If liquid water is present, even intermittently, life forms present would at least be protected from the lethal radiation bombarding the surface. However, life would have to contend with variations in pressure and possibly extended periods of desiccation. The research reported here involves both active metabolism (methanogenesis) at 400 and 50 mbar of pressure, pressures that would be found in the near subsurface of Mars, and survival following desiccation at both 1 bar (a pressure that would be found in the Martian subsurface) and 6 mbar (the lowest pressure at the surface and very near subsurface). The three methanogens tested for active metabolism, Methanothermobacter wolfeii, Methanosarcina barkeri and Methanobacterium formicicum, all demonstrated methane production at both 400 and 50 mbar on JSC Mars-1, a Mars soil simulant. Methane production at 50 mbar was much reduced compared to that at 400 mbar, most likely due to the greater stress at the lower pressure. In desiccation survival experiments, M. barkeri had survived 330 days of desiccation at 1 bar, while M. wolfeii and M. formicicum survived 180 and 120 days, respectively. Methanococcus maripaludis did not survive desiccation at all at 1 bar. At 6 mbar, M. wolfeii, M. barkeri and M. formicicum survived 120 days of desiccation while M. maripaludis survived 60 days. These results along with results from previous research would seem to indicate that there is no reason that methanogens could not inhabit the subsurface of Mars.

  5. Ruminal Fermentation of Anti-Methanogenic Nitrate- and Nitro-Containing Forages In Vitro

    PubMed Central

    Anderson, Robin C.; Ripley, Laura H.; Bowman, Jan G. P.; Callaway, Todd R.; Genovese, Kenneth J.; Beier, Ross C.; Harvey, Roger B.; Nisbet, David J.

    2016-01-01

    Nitrate, 3-nitro-1-propionic acid (NPA) and 3-nitro-1-propanol (NPOH) can accumulate in forages and be poisonous to animals if consumed in high enough amounts. These chemicals are also recognized as potent anti-methanogenic compounds, but plants naturally containing these chemicals have been studied little in this regard. Presently, we found that nitrate-, NPA-, or NPOH-containing forages effectively decreased methane production, by 35–87%, during in vitro fermentation by mixed cultures of ruminal microbes compared to fermentation by cultures incubated similarly with alfalfa. Methane production was further decreased during the incubation of mixed cultures also inoculated with Denitrobacterium detoxificans, a ruminal bacterium known to metabolize nitrate, NPA, and NPOH. Inhibition of methanogens within the mixed cultures was greatest with the NPA- and NPOH-containing forages. Hydrogen accumulated in all the mixed cultures incubated with forages containing nitrate, NPA or NPOH and was dramatically higher, exceeding 40 μmol hydrogen/mL, in mixed cultures incubated with NPA-containing forage but not inoculated with D. detoxificans. This possibly reflects the inhibition of hydrogenase-catalyzed uptake of hydrogen produced via conversion of 50 μmol added formate per milliliter to hydrogen. Accumulations of volatile fatty acids revealed compensatory changes in fermentation in mixed cultures incubated with the nitrate-, NPA-, and NPOH-containing forages as evidenced by lower accumulations of acetate, and in some cases, higher accumulations of butyrate and lower accumulations of ammonia, iso-buytrate, and iso-valerate compared to cultures incubated with alfalfa. Results reveal that nitrate, NPA, and NPOH that accumulate naturally in forages can be made available within ruminal incubations to inhibit methanogenesis. Further research is warranted to determine if diets can be formulated with nitrate-, NPA-, and NPOH-containing forages to achieve efficacious mitigation

  6. Ruminal Fermentation of Anti-Methanogenic Nitrate- and Nitro-Containing Forages In Vitro.

    PubMed

    Anderson, Robin C; Ripley, Laura H; Bowman, Jan G P; Callaway, Todd R; Genovese, Kenneth J; Beier, Ross C; Harvey, Roger B; Nisbet, David J

    2016-01-01

    Nitrate, 3-nitro-1-propionic acid (NPA) and 3-nitro-1-propanol (NPOH) can accumulate in forages and be poisonous to animals if consumed in high enough amounts. These chemicals are also recognized as potent anti-methanogenic compounds, but plants naturally containing these chemicals have been studied little in this regard. Presently, we found that nitrate-, NPA-, or NPOH-containing forages effectively decreased methane production, by 35-87%, during in vitro fermentation by mixed cultures of ruminal microbes compared to fermentation by cultures incubated similarly with alfalfa. Methane production was further decreased during the incubation of mixed cultures also inoculated with Denitrobacterium detoxificans, a ruminal bacterium known to metabolize nitrate, NPA, and NPOH. Inhibition of methanogens within the mixed cultures was greatest with the NPA- and NPOH-containing forages. Hydrogen accumulated in all the mixed cultures incubated with forages containing nitrate, NPA or NPOH and was dramatically higher, exceeding 40 μmol hydrogen/mL, in mixed cultures incubated with NPA-containing forage but not inoculated with D. detoxificans. This possibly reflects the inhibition of hydrogenase-catalyzed uptake of hydrogen produced via conversion of 50 μmol added formate per milliliter to hydrogen. Accumulations of volatile fatty acids revealed compensatory changes in fermentation in mixed cultures incubated with the nitrate-, NPA-, and NPOH-containing forages as evidenced by lower accumulations of acetate, and in some cases, higher accumulations of butyrate and lower accumulations of ammonia, iso-buytrate, and iso-valerate compared to cultures incubated with alfalfa. Results reveal that nitrate, NPA, and NPOH that accumulate naturally in forages can be made available within ruminal incubations to inhibit methanogenesis. Further research is warranted to determine if diets can be formulated with nitrate-, NPA-, and NPOH-containing forages to achieve efficacious mitigation in

  7. Methylmercury decomposition in sediments and bacterial cultures: Involvement of methanogens and sulfate reducers in oxidative demethylation

    USGS Publications Warehouse

    Oremland, R.S.; Culbertson, C.W.; Winfrey, M.R.

    1991-01-01

    Demethylation of monomethylmercury in freshwater and estuarine sediments and in bacterial cultures was investigated with 14CH3HgI. Under anaerobiosis, results with inhibitors indicated partial involvement of both sulfate reducers and methanogens, the former dominating estuarine sediments, while both were active in freshwaters. Aerobes were the most significant demethylators in estuarine sediments, but were unimportant in freshwater sediments. Products of anaerobic demethylation were mainly 14CO2 as well as lesser amounts of 14CH4. Acetogenic activity resulted in fixation of some 14CO2 produced from 14CH3HgI into acetate. Aerobic demethylation in estuarine sediments produced only 14CH4, while aerobic demethylation in freshwater sediments produced small amounts of both 14CH4 and 14CO2. Two species of Desulfovibrio produced only traces of 14CH4 from 14CH3HgI, while a culture of a methylotrophic methanogen formed traces of 14CO2 and 14CH4 when grown on trimethylamine in the presence of the 14CH3HgI. These results indicate that both aerobes and anaerobes demethylate mercury in sediments, but that either group may dominate in a particular sediment type. Aerobic demethylation in the estuarine sediments appeared to proceed by the previously characterized organomercurial-lyase pathway, because methane was the sole product. However, aerobic demethylation in freshwater sediments as well as anaerobic demethylation in all sediments studied produced primarily carbon dioxide. This indicates the presence of an oxidative pathway, possibly one in which methylmercury serves as an analog of one-carbon substrates.

  8. In Vitro Cultivation of Cryptosporidium Species

    PubMed Central

    Arrowood, Michael J.

    2002-01-01

    The in vitro cultivation of protozoan parasites of the genus Cryptosporidium has advanced significantly in recent years. These obligate, intracellular parasites colonize the epithelium of the digestive and respiratory tracts, are often difficult to obtain in significant numbers, produce durable oocysts that defy conventional chemical disinfection methods, and are persistently infectious when stored at refrigerated temperatures (4 to 8°C). While continuous culture and efficient life cycle completion (oocyst production) have not yet been achieved in vitro, routine methods for parasite preparation and cell culture infection and assays for parasite life cycle development have been established. Parasite yields may be limited, but in vitro growth is sufficient to support a variety of research studies, including assessing potential drug therapies, evaluating oocyst disinfection methods, and characterizing life cycle stage development and differentiation. PMID:12097247

  9. 34 CFR 303.15 - Include; including.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... 34 Education 2 2010-07-01 2010-07-01 false Include; including. 303.15 Section 303.15 Education Regulations of the Offices of the Department of Education (Continued) OFFICE OF SPECIAL EDUCATION AND REHABILITATIVE SERVICES, DEPARTMENT OF EDUCATION EARLY INTERVENTION PROGRAM FOR INFANTS AND TODDLERS...

  10. Iron sulfide attenuates the methanogenic toxicity of elemental copper and zinc oxide nanoparticles and their soluble metal ion analogs.

    PubMed

    Gonzalez-Estrella, Jorge; Gallagher, Sara; Sierra-Alvarez, Reyes; Field, Jim A

    2016-04-01

    Elemental copper (Cu(0)) and zinc oxide (ZnO) nanoparticle (NP) toxicity to methanogens has been attributed to the release of soluble metal ions. Iron sulfide (FeS) partially controls the soluble concentration of heavy metals and their toxicity in aquatic environments. Heavy metals displace the Fe from FeS forming poorly soluble metal sulfides in the FeS matrix. Therefore, FeS may be expected to attenuate the NP toxicity. This work assessed FeS as an attenuator of the methanogenic toxicity of Cu(0) and ZnO NPs and their soluble salt analogs. The toxicity attenuation capacity of fine (25-75μm) and coarse (500 to 1200μm) preparations of FeS (FeS-f and FeS-c respectively) was tested in the presence of highly inhibitory concentrations of CuCl2, ZnCl2 Cu(0) and ZnO NPs. FeS-f attenuated methanogenic toxicity better than FeS-c. The results revealed that 2.5× less FeS-f than FeS-c was required to recover the methanogenic activity to 50% (activity normalized to uninhibited controls). The results also indicated that a molar FeS-f/Cu(0) NP, FeS-f/ZnO NP, FeS-f/ZnCl2, and FeS-f/CuCl2 ratio of 2.14, 2.14, 4.28, and 8.56 respectively, was necessary to recover the methanogenic activity to >75%. Displacement experiments demonstrated that CuCl2 and ZnCl2 partially displaced Fe from FeS. As a whole, the results indicate that not all the sulfide in FeS was readily available to react with the soluble Cu and Zn ions which may explain the need for a large stoichiometric excess of FeS to highly attenuate Cu and Zn toxicity. Overall, this study provides evidence that FeS attenuates the toxicity caused by Cu(0) and ZnO NPs and their soluble ion analogs to methanogens. PMID:26803736

  11. Effects of cultivation frequency on sugarcane yields

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Reducing the number of cultivations during one or more years of a four year crop cycle reduces production expenses and could increase profitability if yields are not adversely affected. This study was initiated to determine the effects of cultivation on yields of sugarcane grown on a clay soil both ...

  12. Effects of cultivation frequency on sugarcane yields

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Reducing the number of cultivations during one or more years of a four year crop cycle may potentially reduce production expenses for Louisiana sugarcane growers. This study was initiated to determine the effects of cultivation on yields of sugarcane grown on a clay soil both on an annual basis and ...

  13. Allelopathy as a potential strategy to improve microalgae cultivation

    PubMed Central

    2013-01-01

    One of the main obstacles for continuous productivity in microalgae cultivation is the presence of biological contaminants capable of eliminating large numbers of cells in a matter of days or even hours. However, a number of strategies are being used to combat and prevent contamination in microalgae cultivation. These strategies include the use of extreme conditions in the culture media such as high salinity and high pH to create an unfavorable environment for the competitive organisms or predators of the microalgae. Numerous studies have explored the potential of naturally occurring bioactive secondary metabolites, which are natural products from plants and microorganisms, as a source of such compounds. Some of these compounds are herbicides, and marine and freshwater microalgae are a source of these compounds. Microalgae produce a remarkable diversity of biologically active metabolites. Results based on the allelopathic potential of algae have only been described for laboratory-scale production and not for algae cultivation on a pilot scale. The adoption of allelopathy on microalgal strains is an unexplored field and may be a novel solution to improve algae production. Here we present information showing the diversity of allelochemicals from microalgae and the use of an allelopathic approach to control microalgae cultivation on a pilot scale based on R&D activities being carried out in Brazil for biodiesel production. PMID:24499580

  14. Allelopathy as a potential strategy to improve microalgae cultivation.

    PubMed

    Bacellar Mendes, Leonardo Brantes; Vermelho, Alane Beatriz

    2013-01-01

    One of the main obstacles for continuous productivity in microalgae cultivation is the presence of biological contaminants capable of eliminating large numbers of cells in a matter of days or even hours. However, a number of strategies are being used to combat and prevent contamination in microalgae cultivation. These strategies include the use of extreme conditions in the culture media such as high salinity and high pH to create an unfavorable environment for the competitive organisms or predators of the microalgae. Numerous studies have explored the potential of naturally occurring bioactive secondary metabolites, which are natural products from plants and microorganisms, as a source of such compounds. Some of these compounds are herbicides, and marine and freshwater microalgae are a source of these compounds. Microalgae produce a remarkable diversity of biologically active metabolites. Results based on the allelopathic potential of algae have only been described for laboratory-scale production and not for algae cultivation on a pilot scale. The adoption of allelopathy on microalgal strains is an unexplored field and may be a novel solution to improve algae production. Here we present information showing the diversity of allelochemicals from microalgae and the use of an allelopathic approach to control microalgae cultivation on a pilot scale based on R&D activities being carried out in Brazil for biodiesel production. PMID:24499580

  15. Methyl coenzyme M reductase (mcrA) gene abundance correlates with activity measurements of methanogenic H2/CO2-enriched anaerobic biomass

    PubMed Central

    Morris, Rachel; Schauer-Gimenez, Anne; Bhattad, Ujwal; Kearney, Colleen; Struble, Craig A; Zitomer, Daniel; Maki, James S

    2014-01-01

    Biologically produced methane (CH4) from anaerobic digesters is a renewable alternative to fossil fuels, but digester failure can be a serious problem. Monitoring the microbial community within the digester could provide valuable information about process stability because this technology is dependent upon the metabolic processes of microorganisms. A healthy methanogenic community is critical for digester function and CH4 production. Methanogens can be surveyed and monitored using genes and transcripts of mcrA, which encodes the α subunit of methyl coenzyme M reductase – the enzyme that catalyses the final step in methanogenesis. Using clone libraries and quantitative polymerase chain reaction, we compared the diversity and abundance of mcrA genes and transcripts in four different methanogenic hydrogen/CO2 enrichment cultures to function, as measured by specific methanogenic activity (SMA) assays using H2/CO2. The mcrA gene copy number significantly correlated with CH4 production rates using H2/CO2, while correlations between mcrA transcript number and SMA were not significant. The DNA and cDNA clone libraries from all enrichments were distinctive but community diversity also did not correlate with SMA. Although hydrogenotrophic methanogens dominated these enrichments, the results indicate that this methodology should be applicable to monitoring other methanogenic communities in anaerobic digesters. Ultimately, this could lead to the engineering of digester microbial communities to produce more CH4 for use as renewable fuel. PMID:24320083

  16. The Community Land Model underestimates land-use CO2 emissions by neglecting soil disturbance from cultivation

    NASA Astrophysics Data System (ADS)

    Levis, S.; Hartman, M. D.; Bonan, G. B.

    2014-04-01

    The Community Land Model (CLM) can simulate planting and harvesting of crops but does not include effects of cultivation on soil carbon decomposition. The biogeochemistry model DayCent does account for cultivation and provides a baseline for evaluating the CLM. With the goal of representing cultivation effects on soil carbon decomposition, we implemented the DayCent cultivation parameterization in the CLM and compared CLM and DayCent simulations at eight Midwestern United States sites with and without the cultivation parameterization. Cultivation decreases soil carbon by about 1350 gC m-2 in the CLM and 1660 gC m-2 in DayCent across the eight sites from the first cultivation (early 1900s) to 2010. CLM crop simulations without cultivation have soil carbon gain, not loss, over this period, in contrast to the expected declining trends in agricultural soil carbon. A global cultivation simulation for 1973-2004 reduces ecosystem carbon by 0.4 Pg yr-1 over temperate corn, soybean, and cereal crop areas, which occupy approximately 1/3 of global crop area. Earth System Models may improve their atmospheric CO2 and soil carbon simulations by accounting for enhanced decomposition from cultivation.

  17. Earth System Models that simulate crops underestimate CO2 emissions from land use by neglecting soil disturbance due to cultivation

    NASA Astrophysics Data System (ADS)

    Levis, S.; Hartman, M. D.; Bonan, G. B.

    2013-12-01

    The Community Land Model (CLM) can simulate planting and harvesting of crops but does not include effects of cultivation on soil carbon decomposition. The biogeochemistry model DayCent does account for cultivation and provides a baseline for evaluating the CLM. With the goal of representing cultivation effects on soil carbon decomposition, we implemented the DayCent cultivation parameterization in the CLM and compared CLM and DayCent simulations at eight Midwestern United States sites with and without the cultivation parameterization. Cultivation decreases soil carbon by about 1350 g C m-2 in the CLM and 1660 g C m-2 in DayCent across the eight sites from first cultivation (early 1900s) to 2010. CLM crop simulations without cultivation have soil carbon gain, not loss, over this period, in contrast to the expected declining trends in agricultural soil carbon. A global cultivation simulation for 1973-2004 reduces ecosystem carbon by 0.4 Pg yr-1 over temperate corn, soybean, and cereal crop areas, which occupy approximately 1/3 of global crop area. Earth System Models may improve their atmospheric CO2 and soil carbon simulations by accounting for enhanced decomposition from cultivation.

  18. Comparison of methanogenic community structure and anaerobic process performance treating swine wastewater between pilot and optimized lab scale bioreactors.

    PubMed

    Kim, Woong; Cho, Kyungjin; Lee, Seungyong; Hwang, Seokhwan

    2013-10-01

    To investigate methanogenic community structure and process performance of anaerobic digestion treating swine wastewater at different scale, a pilot plant with 20 m(3) of effective working volume and lab scale methanogenic digester with 6L working volume were operated for 71 days and 6 turnover periods, respectively. During the steady state of anaerobic digestion, COD and VS removal efficiency in pilot plant were 65.3±3.2, 51.6±4.3%, respectively, which was similar to those in lab scale. However, calculated VFAs removal efficiency and methane yield were lower in pilot plant than in lab scale digester. Also, organics removal efficiencies, which consist of total carbohydrates, proteins, and lipids, were different between pilot and lab scale. These results were thought to be due to the ratio of carbohydrates to proteins in the raw swine wastewater. As a result of qualitative microbial analysis, Methanoculleus receptaculii, and Methanoculleus bourgensis, were commonly concerned with methane production. PMID:23489568

  19. Shifts in methanogenic subpopulations measured with antibody probes in a fixed-bed loop anaerobic bioreactor treating sulfite evaporator condensate

    SciTech Connect

    Macario, A.J.L.; de Macario, E.C. ); Ney, U.; Schoberth, S.M.; Sahm, H. )

    1989-08-01

    A fixed-bed loop, high-rate anaerobic bioreactor treating sulfite evaporator condensate was sampled when it reached steady state and afterwards following perturbations during a 14-month period. By using immunotechnology, it was observed that shifts in methanogenic subpopulations occurred in association with perturbations, such as restarting and relocating the biomass into a different tank. Methanogens related to Methanobacterium bryantii MoHG and Methanobrevibacter smithii ALI were numerous throughout the observation period, while Methanosarcina mazei S6 and Methanosarcina thermophila TM1 were found in the early and late samples, respectively. Also, Methanobacterium formicicum was more numerous at the top portion of the bioreactor, while Methanobrevibacter arboriphilus AZ and DC were at the bottom. Sample formalinization required for prolonged storage proved suitable for antigen preservation.

  20. Low-strength ultrasonication positively affects methanogenic granules toward higher AD performance: Implications from microbial community shift.

    PubMed

    Cho, Si-Kyung; Kim, Dong-Hoon; Quince, Christopher; Im, Wan-Taek; Oh, Sae-Eun; Shin, Seung Gu

    2016-09-01

    To elucidate the enhanced methane yield from organic wastes, the effects of low-strength ultrasonication on the microbial community structures in upflow anaerobic sludge blanket reactors were for the first time analyzed using pyrosequencing. Interestingly, a more even microbial community was observed in the ultrasonicated granules than in the control, which could compensate for the decreased richness and resulted in comparable (archaea) or even higher (bacteria) diversity. The ultrasonicated granules contained higher levels of δ-Proteobacteria, of which many are reportedly potential syntrophs, as well as methanogenic genera Methanosaeta, Methanotorris, and Methanococcus. The increased presence of syntrophic bacteria with their methanogenic partners was discussed with respect to hydrogen flux; their selective proliferation seems to be responsible for the enhanced anaerobic performance. This study is the first research shedding light on the novel function of low-strength ultrasound shifting the microbial structure towards better biogas production performance, and will facilitate application of low-strength ultrasound to other bioprocesses. PMID:27150761

  1. Complete genome sequence of the hyperthermophilic methanogen Methanocaldococcus bathoardescens JH146(T) isolated from the basalt subseafloor.

    PubMed

    Kim, You-Tae; Jung, Jong-Hyun; Stewart, Lucy C; Kwon, Soon-Wo; Holden, James F; Park, Cheon-Seok

    2015-12-01

    Methanocaldococcus bathoardescens JH146(T) is a hyperthermophilic and obligate hydrogenotrophic methanogen isolated from low-temperature (26 °C) hydrothermal vent fluid at Axial Seamount in the northeastern Pacific Ocean. It is most closely related to the N2-fixing methanogen Methanocaldococcus sp. FS406-22; however, they differ in that JH146 cannot fix N2 or reductively assimilate nitrate. In this study, we present the complete genome sequence of strain JH146(T) (1,607,556 bp) with its 1635 protein coding genes, and 41 RNA genes. Our analysis focuses on its methane production via the acetyl-CoA pathway and its deleted gene clusters related to nitrogen assimilation. This study extends our understanding of methanogenesis at high temperatures and the impact of these organisms on the biogeochemistry of subseafloor hydrothermal environments and the deep sea. PMID:26094864

  2. Lipid Biomarkers for Methanogens in Hypersaline Cyanobacterial Mats for Guerrero Negro, Baja California Sur

    NASA Technical Reports Server (NTRS)

    Jahnke, Linda L.; Embaye, Tsegereda; Summons, Roger E.; Fonda, Mark (Technical Monitor)

    2002-01-01

    Analyses of sediments from the vicinity of active methane seeps have uncovered a particular suite of lipid biomarker patterns that characterize methane consuming archaea and their syntrophic, sulfate reducing partners. These isoprenoid biomarkers, largely identified by their anomalously light carbon isotopic signatures, have been a topic of intense research activity and are recorded in numerous methane-rich environments from Holocene to Cenozoic. This phenomenon has implications for depleted kerogens at 2.7 Ga on early Earth (Hinrichs 2002). In contrast, the lipid biosignatures of methane producing archaea are not readily identified through distinct isotopic labels and have received comparably little attention in analyses of archaea in environmental samples. Indeed, environmental analyses generally detect only free archaeal lipids, not the intact, polar molecules found in the membrane of living organisms. As part of the Ames NAI, the 'Early Microbial Ecosystem Research Group' (EMERG) is working to understand microbial processes in the hypersaline cyanobacterial mats growing in the salt evaporation ponds of the Exportadora de Sal at Guerrero Negro, Baja California Sur, Mexico. The aim of this study was to develop methods by which we could identify the organisms responsible for methane generation in this environment. While the ester-bound fatty acids, hopanoids and wax esters provide a means to identify most of the bacterial components of these mats, the archaea which Ere evidently present through genomic assays and the fact of intense methane production (Hoehler et al. 200l), have not been identified through their corresponding lipid signatures. Archaeal core lipids present a number of analytical challenges. The core lipids of methanogens comprise C20, C40 and sometimes C25 isoprenoid chains, linked through ether bonds to glycerol. As well as archaeal (C20), sn-2- and sn-3-hydroxyarchaeol are associated particularly with methylotrophic methanogens. Recently, we have

  3. An Overview of Talent Cultivation Models in Foreign Vocational Colleges

    ERIC Educational Resources Information Center

    Wang, Youhua

    2010-01-01

    A talent cultivation model refers to a model of the construction and operation of the cultivation process adopted by a school to achieve its cultivation objectives. Four models, with their differences and similarities, are introduced in this paper.

  4. Examination of the rumen bacteria and methanogenic archaea of wild impalas (Aepyceros melampus melampus) from Pongola, South Africa.

    PubMed

    Cersosimo, Laura M; Lachance, Hannah; St-Pierre, Benoit; van Hoven, Wouter; Wright, André-Denis G

    2015-04-01

    Although the rumen microbiome of domesticated ruminants has been evaluated, few studies have explored the rumen microbiome of wild ruminants, and no studies have identified the rumen microbiome in the impala (Aepyceros melampus melampus). In the present study, next-generation sequencing and real-time polymerase chain reaction were used to investigate the diversity and density of the bacteria and methanogenic archaea residing in the rumen of five adult male impalas, culled during the winter dry season in Pongola, South Africa. A total of 15,323 bacterial 16S rRNA gene sequences (from five impala), representing 3,892 different phylotypes, were assigned to 1,902 operational taxonomic units (OTUs). A total of 20,124 methanogen 16S rRNA gene sequence reads (from four impala), of which 5,028 were unique, were assigned to 344 OTUs. From the total sequence reads, Bacteroidetes, Proteobacteria, and Firmicutes were the most abundant bacterial phyla. While the majority of the bacterial genera found were unclassified, Prevotella and Cupriavidus were the most abundant classified genera. For methanogens, the genera Methanobrevibacter and Methanosphaera represented 94.3% and 4.0% of the classified sequences, respectively. Most notable was the identification of Methanobrevibacter thaueri-like 16S rRNA gene sequence reads in all four impala samples, representing greater than 30% of each individual's total sequences. Both data sets are accessible through NCBI's Sequence Read Archive (SRA), under study accession number SRP [048619]. The densities of bacteria (1.26 × 10(10)-3.82 × 10(10) cells/ml whole rumen contents) and methanogens (4.48 × 10(8)-7.2 × 10(9) cells/ml of whole rumen contents) from five individual impala were similar to those typically observed in domesticated ruminants. PMID:25351144

  5. Metabolism of Reduced Methylated Sulfur Compounds in Anaerobic Sediments and by a Pure Culture of an Estuarine Methanogen

    PubMed Central

    Kiene, Ronald P.; Oremland, Ronald S.; Catena, Anthony; Miller, Laurence G.; Capone, Douglas G.

    1986-01-01

    Addition of dimethylsulfide (DMS), dimethyldisulfide (DMDS), or methane thiol (MSH) to a diversity of anoxic aquatic sediments (e.g., fresh water, estuarine, alkaline/hypersaline) stimulated methane production. The yield of methane recovered from DMS was often 52 to 63%, although high concentrations of DMS (as well as MSH and DMDS) inhibited methanogenesis in some types of sediments. Production of methane from these reduced methylated sulfur compounds was blocked by 2-bromoethanesulfonic acid. Sulfate did not influence the metabolism of millimolar levels of DMS, DMDS, or MSH added to sediments. However, when DMS was added at ∼2-μM levels as [14C]DMS, metabolism by sediments resulted in a 14CH4/14CO2 ratio of only 0.06. Addition of molybdate increased the ratio to 1.8, while 2-bromoethanesulfonic acid decreased it to 0, but did not block 14CO2 production. These results indicate the methanogens and sulfate reducers compete for DMS when it is present at low concentrations; however, at high concentrations, DMS is a “noncompetitive” substrate for methanogens. Metabolism of DMS by sediments resulted in the appearance of MSH as a transient intermediate. A pure culture of an obligately methylotrophic estuarine methanogen was isolated which was capable of growth on DMS. Metabolism of DMS by the culture also resulted in the transient appearance of MSH, but the organism could grow on neither MSH nor DMDS. The culture metabolized [14C]-DMS to yield a 14CH4/14CO2 ratio of ∼2.8. Reduced methylated sulfur compounds represent a new class of substrates for methanogens and may be potential precursors of methane in a variety of aquatic habitats. PMID:16347202

  6. Aceticlastic and NaCl-Requiring Methanogen “Methanosaeta pelagica” sp. nov., Isolated from Marine Tidal Flat Sediment

    PubMed Central

    Iino, Takao; Suzuki, Ken-Ichiro; Yamaguchi, Kaoru; Kamagata, Yoichi

    2012-01-01

    Among methanogens, only 2 genera, Methanosaeta and Methanosarcina, are known to contribute to methanogenesis from acetate, and Methanosaeta is a specialist that uses acetate specifically. However, Methanosaeta strains so far have mainly been isolated from anaerobic digesters, despite the fact that it is widespread, not only in anaerobic methanogenic reactors and freshwater environments, but also in marine environments, based upon extensive 16S rRNA gene-cloning analyses. In this study, we isolated an aceticlastic methanogen, designated strain 03d30qT, from a tidal flat sediment. Phylogenetic analyses based on 16S rRNA and mcrA genes revealed that the isolate belongs to the genus Methanosaeta. Unlike the other known Methanosaeta species, this isolate grows at Na+ concentrations of 0.20 to 0.80 M, with an optimum concentration of 0.28 M. Quantitative estimation using real-time PCR detected the 16S rRNA gene of the genus Methanosaeta in the marine sediment, and relative abundance ranged from 3.9% to 11.8% of the total archaeal 16S rRNA genes. In addition, the number of Methanosaeta organisms increased with increasing depth and was much higher than that of Methanosarcina organisms, suggesting that aceticlastic methanogens contribute to acetate metabolism to a greater extent than previously thought in marine environments, where sulfate-reducing acetate oxidation prevails. This is the first report on marine Methanosaeta species, and based on phylogenetic and characteristic studies, the name “Methanosaeta pelagica” sp. nov. is proposed for this novel species, with type strain 03d30q. PMID:22344667

  7. Methanogens and Methanogenesis in the Rumens and Ceca of Lambs Fed Two Different High-Grain-Content Diets

    PubMed Central

    Morgavi, D. P.; Martin, C.

    2013-01-01

    The amount and nature of dietary starch are known to influence the extent and site of feed digestion in ruminants. However, how starch degradability may affect methanogenesis and methanogens along the ruminant's digestive tract is poorly understood. This study examined the diversity and metabolic activity of methanogens in the rumen and cecum of lambs receiving wheat or corn high-grain-content diets. Methane production in vivo and ex situ was also monitored. In vivo daily methane emissions (CH4 g/day) were 36% (P < 0.05) lower in corn-fed lambs than in wheat-fed lambs. Ex situ methane production (μmol/h) was 4-fold higher for ruminal contents than for cecal contents (P < 0.01), while methanogens were 10-fold higher in the rumen than in the cecum (mcrA copy numbers; P < 0.01). Clone library analysis indicated that Methanobrevibacter was the dominant genus in both sites. Diet induced changes at the species level, as the Methanobrevibacter millerae-M. gottschalkii-M. smithii clade represented 78% of the sequences from the rumen of wheat-fed lambs and just about 52% of the sequences from the rumen of the corn-fed lambs. Diet did not affect mcrA expression in the rumen. In the cecum, however, expression was 4-fold and 2-fold lower than in the rumen for wheat- and corn-fed lambs, respectively. Though we had no direct evidence for compensation of reduced rumen methane production with higher cecum methanogenesis, the ecology of methanogens in the cecum should be better considered. PMID:23241983

  8. Methanogenic community change in a full-scale UASB reactor operated at a low F/M ratio.

    PubMed

    Kolukirik, M; Ince, O; Ince, B Kasapgil

    2007-06-01

    A full-scale upflow anaerobic sludge blanket (UASB) reactor was investigated in terms of archaeal composition, acetoclastic methanogenic capacity and performance over a 2-year period. Performance of the reactor in terms of COD removal efficiency varied between 60% and 80% at organic loading rates (OLRs) in the range of 2.5-12 kg COD m-3 d-1. The reactor had been operated under a F/M (food to microorganisms) ratio of 0.02-0.03 gCOD gTVS-1 d-1, which is much lower than the typical values reported for similar reactors. According to specific methanogenic activity (SMA) tests the anaerobic sludge was operating at only 12-34% of its potential acetoclastic methanogenic capacity. These results demonstrated that the UASB reactor was under loaded compared to its maximum loading capacity. All other operational parameters had been maintained within their desired ranges. The SMA test and Fluorescence in situ hybridization (FISH) results revealed that a decrease in the acetoclastic methanogenic activity of the UASB sludge from 344 mL CH4 gTVS-1 d-1 to 109 mL CH4 gTVS-1 d-1 coincided with a decrease in the relative abundance of acetoclastic Methanosaeta from 90%+/-1.2 to 79%+/-1.4 of the archaeal population, and an increase in the relative abundance of hydrogenotrophic Methanobacteriales from non-detectable levels to 24%+/-0.7% of the archaeal population during the 2-year operation of the reactor. The relative abundance of archaeal cells within the UASB sludge was in the range of 15-17%. PMID:17558771

  9. Metabolism of reduced methylated sulfur compounds in anaerobic sediments and by a pure culture of an estuarine methanogen

    USGS Publications Warehouse

    Kiene, R.P.; Oremland, Ronald S.; Catena, Anthony; Miller, Laurence G.; Capone, D.G.

    1986-01-01

    Addition of dimethylsulfide (DMS), dimethyldisulfide (DMDS), or methane thiol (MSH) to a diversity of anoxic aquatic sediments (e.g., fresh water, estuarine, alkaline/hypersaline) stimulated methane production. The yield of methane recovered from DMS was often 52 to 63%, although high concentrations of DMS (as well as MSH and DMDS) inhibited methanogenesis in some types of sediments. Production of methane from these reduced methylated sulfur compounds was blocked by 2-bromoethanesulfonic acid. Sulfate did not influence the metabolism of millimolar levels of DMS, DMDS, or MSH added to sediments. However, when DMS was added at ∼2-μM levels as [14C]DMS, metabolism by sediments resulted in a 14CH4/14CO2 ratio of only 0.06. Addition of molybdate increased the ratio to 1.8, while 2-bromoethanesulfonic acid decreased it to 0, but did not block 14CO2 production. These results indicate the methanogens and sulfate reducers compete for DMS when it is present at low concentrations; however, at high concentrations, DMS is a “noncompetitive” substrate for methanogens. Metabolism of DMS by sediments resulted in the appearance of MSH as a transient intermediate. A pure culture of an obligately methylotrophic estuarine methanogen was isolated which was capable of growth on DMS. Metabolism of DMS by the culture also resulted in the transient appearance of MSH, but the organism could grow on neither MSH nor DMDS. The culture metabolized [14C]-DMS to yield a 14CH4/14CO2 ratio of ∼2.8. Reduced methylated sulfur compounds represent a new class of substrates for methanogens and may be potential precursors of methane in a variety of aquatic habitats.

  10. Three stages MBR (methanogenic, aerobic biofilm and membrane filtration) for the treatment of low-strength wastewaters.

    PubMed

    Buntner, D; Sánchez Sánchez, A; Garrido, J M

    2011-01-01

    The use of a new three stages MBR process with a first methanogenic UASB stage, a second stage with aerobic biofilm growing on small carrier elements maintained in suspension and third stage with membrane filtration module is presented. The objective of the first methanogenic chamber is to diminish COD of the raw wastewater, producing a biogas rich in methane, and decrease the sludge production. In the second stage, the remaining soluble biodegradable COD is oxidized by heterotrophs. In the third stage, the membrane modules could be operated at higher fluxes than those reported for AnMBR systems, and similar to those obtained in aerobic MBRs. In this sense, the concept of these three stages MBR is to join the advantages of the methanogenic and aerobic membrane bioreactor processes, by reducing energy requirements for aeration, producing biogas with high methane percentage and a permeate with very low COD content. A synthetic wastewater was fed to the three stages MBR. COD in the influent was between 200 and 1,200 mg/L, ammonium ranged from 10 to 35 mg/L and phosphorous concentration was 8 mg/L. OLR in-between 1 and 3 kg COD/(m3 d) and a HRT of 13-21 h were applied. Temperature was between 17.5 and 23.2 degrees C. During the whole operating period the COD removal efficiencies were in the range of 90 and 96% of which in between 40 and 80% was removed in the first methanogenic chamber. Biogas production with methane content between 75 and 80% was observed. With regard to membrane operation, average permeabilities around 150 L/(m2 h bar) were achieved, operating with fluxes of 11-15 L/(m2 h). PMID:22097013

  11. Methanomethylovorans uponensis sp. nov., a methylotrophic methanogen isolated from wetland sediment.

    PubMed

    Cha, In-Tae; Min, Ui-Gi; Kim, So-Jeong; Yim, Kyung June; Roh, Seong Woon; Rhee, Sung-Keun

    2013-12-01

    A novel mesophilic, methylotrophic, methanogenic archaeon, designated strain EK1(T), was enriched and isolated from wetland sediment. Phylogenetic analysis showed that strain EK1(T) was affiliated with the genus Methanomethylovorans within the family Methanosarcinaceae, and shared the highest 16S rRNA and methyl-coenzyme M reductase alpha-subunit gene sequence similarity with the type strain of Methanomethylovorans hollandica (98.8 and 92.6 %, respectively). The cells of strain EK1(T) were observed to be Gram-negative, non-motile and irregular cocci that did not lyse in 0.1 % (w/v) sodium dodecyl sulfate. Methanol, mono-, di- and trimethylamine, dimethyl sulfide and methanethiol were found to be used as catabolic and methanogenic substrates, whereas H2/CO2, formate, 2-propanol and acetate were not. Growth was observed at 25-40 °C (optimum, 37 °C), at pH 5.5-7.5 (optimum, pH 6.0-6.5) and in the presence of 0-0.1 M NaCl (optimum, 0 M). Growth and methane production rates were stimulated in the presence of H2/CO2 although methane production and growth yields were not significantly affected; acetate, formate, 2-propanol and CO/CO2/N2 did not affect methane production. CoCl2 (0.6-2.0 μM) and FeCl2 (25 mg/l) stimulated growth, while yeast extract and peptone did not. The DNA-DNA hybridization experiment revealed a relatedness of <20 % between EK1(T) and the type strains of the genus Methanomethylovorans. The DNA G+C content of strain EK1(T) was determined to be 39.2 mol%. Based on the polyphasic taxonomic study, strain EK1(T) represents a novel species belonging to the genus Methanomethylovorans, for which the name Methanomethylovorans uponensis sp. nov. is proposed. The type strain is strain EK1(T)(=NBRC 109636(T) = KCTC 4119(T) = JCM 19217(T)). PMID:24000091

  12. Characterization of metabolic performance of methanogenic granules treating brewery wastewater: role of sulfate-reducing bacteria.

    PubMed Central

    Wu, W M; Hickey, R F; Zeikus, J G

    1991-01-01

    the metabolism of H2, formate, and acetate, but ethanol and propionate were converted via sulfate reduction by approximately 28 and 60%, respectively. In the presence of 2.0 mM molybdate, syntrophic propionate and ethanol conversion by the granules was inhibited by 97 and 29%, respectively. The data show that in this granular microbial consortium, methanogens and sulfate-reducing bacteria did not compete for common substrates. Syntrophic propionate and ethanol conversion was likely performed primarily by sulfate-reducing bacteria, while H2, formate, and acetate were consumed primarily by methanogens. Images PMID:1785921

  13. Long-Term Incubation Reveals Methanogenic Biodegradation of C5 and C6 iso-Alkanes in Oil Sands Tailings.

    PubMed

    Siddique, Tariq; Mohamad Shahimin, Mohd Faidz; Zamir, Saima; Semple, Kathleen; Li, Carmen; Foght, Julia M

    2015-12-15

    iso-Alkanes are major components of petroleum and have been considered recalcitrant to biodegradation under methanogenic conditions. However, indigenous microbes in oil sands tailings ponds exposed to solvents rich in 2-methylbutane, 2-methylpentane, 3-methylpentane, n-pentane, and n-hexane produce methane in situ. We incubated defined mixtures of iso- or n-alkanes with mature fine tailings from two tailings ponds of different ages historically exposed to different solvents: one, ~10 years old, receiving C5-C6 paraffins and the other, ~35 years old, receiving naphtha. A lengthy incubation (>6 years) revealed iso-alkane biodegradation after lag phases of 900-1800 and ~280 days, respectively, before the onset of methanogenesis, although lag phases were shorter with n-alkanes (~650-1675 and ~170 days, respectively). 2-Methylpentane and both n-alkanes were completely depleted during ~2400 days of incubation, whereas 2-methylbutane and 3-methylpentane were partially depleted only during active degradation of 2-methylpentane, suggesting co-metabolism. In both cases, pyrotag sequencing of 16S rRNA genes showed codominance of Peptococcaceae with acetoclastic (Methanosaeta) and hydrogenotrophic (Methanoregula and Methanolinea) methanogens. These observations are important for predicting long-term greenhouse-gas emissions from oil sands tailings ponds and extend the known range of hydrocarbons susceptible to methanogenic biodegradation in petroleum-impacted anaerobic environments. PMID:26571341

  14. Impact of temperature, microwave radiation and organic loading rate on methanogenic community and biogas production during fermentation of dairy wastewater.

    PubMed

    Zielińska, Magdalena; Cydzik-Kwiatkowska, Agnieszka; Zieliński, Marcin; Dębowski, Marcin

    2013-02-01

    This study analyzed dairy wastewater fermentation in convection- and microwave-heated hybrid reactors at loadings of 1 and 2 kg COD/(m3 d) and temperatures of 35 and 55 °C. The biomass was investigated at a molecular level to determine the links between the operational parameters of anaerobic digestion and methanogenic Archaea structure. The highest production of biogas with methane content of ca. 67% was noted in the mesophilic microwave-heated reactors. The production of methane-rich biogas and the overall diversity of Archaea was determined by Methanosarcinaceae presence. The temperature and the application of microwaves were the main factors explaining the variations in the methanogen community. At 35 °C, the microwave heating stimulated the growth of highly diverse methanogen assemblages, promoting Methanosarcina barkeri presence and excluding Methanosarcina harudinacea from the biomass. A temperature increase to 55 °C lowered Methanosarcinaceae abundance and induced a replacement of Methanoculleus palmolei by Methanosarcina thermophila. PMID:23262005

  15. A Reactive-Transport Model Describing Methanogen Growth and Methane Production in Diffuse Flow Vents at Axial Seamount

    NASA Astrophysics Data System (ADS)

    Algar, C. K.

    2015-12-01

    Hydrogenotrophic methanogenesis is an important mode of metabolism in deep-sea hydrothermal vents. Diffuse vent fluids often show a depletion in hydrogen with a corresponding increase in methane relative to pure-mixing of end member fluid and seawater, and genomic surveys show an enrichment in genetic sequences associated with known methanogens. However, because we cannot directly sample the subseafloor habitat where these organisms are living, constraining the size and activity of these populations remains a challenge and limits our ability to quantify the role they play in vent biogeochemistry. Reactive-transport modeling may provide a useful tool for approaching this problem. Here we present a reactive-transport model describing methane production along the flow-path of hydrothermal fluid from its high temperature end-member to diffuse venting at the seafloor. The model is set up to reflect conditions at several diffuse vents in the Axial Seamount. The model describes the growth of the two dominant thermophilic methanogens, Methanothermococcus and Methanocaldococcus, observed at Axial seamount. Monod and Arrhenius constants for Methanothermococcus thermolithotrophicus and Methanocaldococcus jannaschii were obtained for the model using chemostat and bottle experiments at varying temperatures. The model is used to investigate the influence of different mixing regimes on the subseafloor populations of these methanogens. By varying the model flow path length and subseafloor cell concentrations, and fitting to observed hydrogen and methane concentrations in the venting fluid, the subseafloor biomass, fluid residence time, and methane production rate can be constrained.

  16. Forensic fingerprinting of oil-spill hydrocarbons in a methanogenic environment-Mandan, ND and Bemidji, MN

    USGS Publications Warehouse

    Hostettler, F.D.; Wang, Y.; Huang, Y.; Cao, W.; Bekins, B.A.; Rostad, C.E.; Kulpa, C.F.; Laursen, A.

    2007-01-01

    In recent decades forensic fingerprinting of oil-spill hydrocarbons has emerged as an important tool for correlating oils and for evaluating their source and character. Two long-term hydrocarbon spills, an off-road diesel spill (Mandan, ND) and a crude oil spill (Bemidji, MN) experiencing methanogenic biodegradation were previously shown to be undergoing an unexpected progression of homologous n-alkane and n-alkylated cyclohexane loss. Both exhibited degradative losses proceeding from the high-molecular-weight end of the distributions, along with transitory concentration increases of lower-molecular-weight homologs. Particularly in the case of the diesel fuel spill, these methanogenic degradative patterns can result in series distributions that mimic lower cut refinery fuels or admixture with lower cut fuels. Forensic fingerprinting in this long-term spill must therefore rely on more recalcitrant series, such as polycyclic aromatic hydrocarbon or drimane sesquiterpane profiles, to prove if the spilled oil is single-sourced or whether there is verifiable admixture with other extraneous refinery fuels. Degradation processes impacting n-alkanes and n-alkylated ring compounds, which make these compounds unsuitable for fingerprinting, nevertheless are of interest in understanding methanogenic biodegradation. Copyright ?? Taylor & Francis Group, LLC.

  17. Methanogenic population dynamics and performance of an anaerobic membrane bioreactor (AnMBR) treating swine manure under high shear conditions.

    PubMed

    Padmasiri, Sudini I; Zhang, Jiangzhao; Fitch, Mark; Norddahl, Birgir; Morgenroth, Eberhard; Raskin, Lutgarde

    2007-01-01

    A 6-L, completely mixed anaerobic bioreactor with an external ultrafiltration membrane module was operated for 300 days to evaluate the startup and performance of an anaerobic membrane bioreactor (AnMBR) treating swine manure. The reactor had a successful startup at the initial loading rate of 1g volatile solids (VS)/L/day. After a two-fold increase in loading rate followed by a sudden, two-fold increase in flow velocity through the membrane module on day 75, the performance of the AnMBR deteriorated as measured by volatile fatty acid (VFA) accumulation, decrease in pH, and decrease in biogas production. The methanogenic population dynamics in the reactor were monitored with terminal restriction fragment length polymorphism (T-RFLP). Changes in the relative levels of Methanosarcinaceae and Methanosaetaceae were consistent with changes in VFA concentrations, i.e., high and low levels of acetate corresponded to a high abundance of Methanosarcinaceae and Methanosaetaceae, respectively. The levels of hydrogenotrophic methanogens of the order of Methanomicrobiales increased during decreased reactor performance suggesting that syntrophic interactions involving hydrogenotrophic methanogens remained intact regardless of the degree of shear in the AnMBR. PMID:17109913

  18. Warmer temperature accelerates methane emissions from the Zoige wetland on the Tibetan Plateau without changing methanogenic community composition

    NASA Astrophysics Data System (ADS)

    Cui, Mengmeng; Ma, Anzhou; Qi, Hongyan; Zhuang, Xuliang; Zhuang, Guoqiang; Zhao, Guohui

    2015-06-01

    Zoige wetland, locating on the Tibet Plateau, accounts for 6.2% of organic carbon storage in China. However, the fate of the organic carbon storage in the Zoige wetland remains poorly understood despite the Tibetan Plateau is very sensitive to global climate change. As methane is an important greenhouse gas and methanogenesis is the terminal step in the decomposition of organic matter, understanding how methane emissions from the Zoige wetland is fundamental to elucidate the carbon cycle in alpine wetlands responding to global warming. In this study, microcosms were performed to investigate the effects of temperature and vegetation on methane emissions and microbial processes in the Zoige wetland soil. A positive correlation was observed between temperature and methane emissions. However, temperature had no effect on the main methanogenic pathway—acetotrophic methanogenesis. Moreover, methanogenic community composition was not related to temperature, but was associated with vegetation, which was also involved in methane emissions. Taken together, these results indicate temperature increases methane emissions in alpine wetlands, while vegetation contributes significantly to methanogenic community composition and is associated with methane emissions. These findings suggest that in alpine wetlands temperature and vegetation act together to affect methane emissions, which furthers a global warming feedback loop.

  19. Isolation of a methanogenic bacterium, Methanosarcina sp. strain FR, for its ability to degrade high concentration of perchloroethylene.

    PubMed

    Cabirol, N; Villemur, R; Perrier, J; Jacob, F; Fouillet, B; Chambon, P

    1998-12-01

    Tetrachloroethylene (PCE) is a toxic compound essentially used as a degreasing and dry-cleaning solvent. A methanogenic and sulfate-reducing consortium that dechlorinates and mineralizes high concentrations of PCE was derived from anaerobically digested sludge obtained from a waste water treatment plant (Bourg-en-Bresse, France). A methanogenic bacterium, strain FR, was isolated from this acclimated consortium. On the basis of morphological and physiological characteristics, strain FR was classified in the genus of Methanosarcina. Phylogeny analysis with the 16S rRNA gene sequence revealed that strain FR is highly related to Methanosarcina mazei and Methanosarcina frisia (99.6 and 99.5% identity, respectively). High concentrations (50-87 microM) of PCE were completely dechlorinated by strain FR cultures at the rate of 76 nM-mg protein(-1).day(-1). PCE dechlorination produced a nonidentified compound. The tracer experiments with [13C]PCE revealed that the product was nonchlorinated. Dechlorination of PCE to trichloroethylene was still active in the presence of boiled cell extract of the strain FR. However, no further dechlorination was observed. This result suggests that a cofactor rather than an enzymatic system is responsible for the first dechlorination of PCE. Dechlorination-active fractions purified from cell extracts on a XAD-4 column revealed the presence of F(420), F(430), and cobamides cofactors. This is the first report of the isolation of a methanogenic bacterium with the ability to dechlorinate high concentrations of PCE to a nonchlorinated product. PMID:10383226

  20. ESA Experiments with the European Modular Cultivation System (EMCS)

    NASA Astrophysics Data System (ADS)

    Brillouet, Claude; Briganti, Luca; Schwarzwalder, Achim

    2008-06-01

    The European Modular Cultivation System (EMCS) is an ESA developed facility dedicated to gravitational biology and especially to plant research. However, experiments using small animals, like insects and small invertebrates are also possible. EMCS is onboard the International Space Station (ISS) since July 2006 and four experiments, including two from ESA, have been already performed. Several others are in their final development phase and shall be flown within the next following years.

  1. Characterization of Persistent Virus-Like Particles in Two Acetate-Fed Methanogenic Reactors

    PubMed Central

    Chien, I-Chieh; Meschke, John Scott; Gough, Heidi L.; Ferguson, John F.

    2013-01-01

    The objective of this study was to characterize the morphology, size-distribution, concentration and genome size of virus-like particles (VLPs) in two acetate-fed Methanosaeta-dominated reactors to better understand the possible correlation between viruses and archaeal hosts. The study reactors were dominated by a single genus of acetoclastic methanogen, Methanosaeta, which was present at 6 to 13 times higher than the combined bacterial populations consisting of Proteobacteria, Firmicutes, and Bacteroidetes. Epifluorescent microscopy showed VLPs concentration of 7.1 ± 1.5×107 VLPs/ml and 8.4 ± 4.3×107 VLPs/ml in the two laboratory reactors. Observations of no detectable import of VLPs with the reactor feed combined long operational time since the last inocula were introduced suggests that the VLP populations were actively propagating in the reactors. Transmission electron microscopy images showed VLPs with morphology consistent with Siphoviridae in both reactors, and VLPs with morphologies consistent with Myoviridae in one of the reactors. The morphology, size-distribution and genome size of VLPs were distinct between reactors suggesting that unique viral populations inhabited each reactor, though the hosts of these VLPs remain unclear. PMID:24278372

  2. Molecular characterization of methanogenic N(5)-methyl-tetrahydromethanopterin: Coenzyme M methyltransferase.

    PubMed

    Upadhyay, Vikrant; Ceh, Katharina; Tumulka, Franz; Abele, Rupert; Hoffmann, Jan; Langer, Julian; Shima, Seigo; Ermler, Ulrich

    2016-09-01

    Methanogenic archaea share one ion gradient forming reaction in their energy metabolism catalyzed by the membrane-spanning multisubunit complex N(5)-methyl-tetrahydromethanopterin: coenzyme M methyltransferase (MtrABCDEFGH or simply Mtr). In this reaction the methyl group transfer from methyl-tetrahydromethanopterin to coenzyme M mediated by cobalamin is coupled with the vectorial translocation of Na(+) across the cytoplasmic membrane. No detailed structural and mechanistic data are reported about this process. In the present work we describe a procedure to provide a highly pure and homogenous Mtr complex on the basis of a selective removal of the only soluble subunit MtrH with the membrane perturbing agent dimethyl maleic anhydride and a subsequent two-step chromatographic purification. A molecular mass determination of the Mtr complex by laser induced liquid bead ion desorption mass spectrometry (LILBID-MS) and size exclusion chromatography coupled with multi-angle light scattering (SEC-MALS) resulted in a (MtrABCDEFG)3 heterotrimeric complex of ca. 430kDa with both techniques. Taking into account that the membrane protein complex contains various firmly bound small molecules, predominantly detergent molecules, the stoichiometry of the subunits is most likely 1:1. A schematic model for the subunit arrangement within the MtrABCDEFG protomer was deduced from the mass of Mtr subcomplexes obtained by harsh IR-laser LILBID-MS. PMID:27342374

  3. High-Pressure Equipment for Growing Methanogenic Microorganisms on Gaseous Substrates at High Temperature

    PubMed Central

    Bernhardt, Günther; Jaenicke, Rainer; Lüdemann, Hans-Dietrich

    1987-01-01

    High-pressure, high-temperature investigations on thermophilic microorganisms that grow on hydrogen or other gaseous substrates require instrumentation which provides sufficient substrate for cell proliferation up to 2 × 108 to 3 × 108 cells per ml under isothermal and isobaric conditions. To minimize H2 leakage and to optimize reproducibility at high pressure and high temperature, 10-ml nickel tubes with a liquid/gas ratio of 1:2 were used in a set of autoclaves connected in series. By applying a hydraulic pump and a 2.5-kW heating device, fast changes in temperature (up to 400°C) and pressure (up to 400 MPa) can be accomplished within less than 10 min. To quantify bacterial growth, determinations of cell numbers per unit volume yielded optimum accuracy. Preliminary experiments with the thermophilic, methanogenic archaebacterium Methanococcus thermolithotrophicus showed that bacterial growth depends on both temperature and pressure. At the optimum temperature, increased hydrostatic pressure up to 50 MPa enhanced the growth yield; at a pressure of >75 MPa, cell lysis dominated. Changes in cell proliferation were accompanied by changes in morphology. PMID:16347413

  4. Anaerobic degradation of toluene and o-xylene by a methanogenic consortium.

    PubMed Central

    Edwards, E A; Grbić-Galić, D

    1994-01-01

    Toluene and o-xylene were completely mineralized to stoichiometric amounts of carbon dioxide, methane, and biomass by aquifer-derived microorganisms under strictly anaerobic conditions. The source of the inoculum was creosote-contaminated sediment from Pensacola, Fla. The adaptation periods before the onset of degradation were long (100 to 120 days for toluene degradation and 200 to 255 days for o-xylene). Successive transfers of the toluene- and o-xylene-degrading cultures remained active. Cell density in the cultures progressively increased over 2 to 3 years to stabilize at approximately 10(9) cells per ml. Degradation of toluene and o-xylene in stable mixed methanogenic cultures followed Monod kinetics, with inhibition noted at substrate concentrations above about 700 microM for o-xylene and 1,800 microM for toluene. The cultures degraded toluene or o-xylene but did not degrade m-xylene, p-xylene, benzene, ethylbenzene, or naphthalene. The degradative activity was retained after pasteurization or after starvation for 1 year. Degradation of toluene and o-xylene was inhibited by the alternate electron acceptors oxygen, nitrate, and sulfate. Degradation was also inhibited by the addition of preferred substrates such as acetate, H2, propionate, methanol, acetone, glucose, amino acids, fatty acids, peptone, and yeast extract. These data suggest that the presence of natural organic substrates or contaminants may inhibit anaerobic degradation of pollutants such as toluene and o-xylene at contaminated sites. Images PMID:8117084

  5. Micro-scale H2–CO2 Dynamics in a Hydrogenotrophic Methanogenic Membrane Reactor

    PubMed Central

    Garcia-Robledo, Emilio; Ottosen, Lars D. M.; Voigt, Niels V.; Kofoed, M. W.; Revsbech, Niels P.

    2016-01-01

    Biogas production is a key factor in a sustainable energy supply. It is possible to get biogas with very high methane content if the biogas reactors are supplied with exogenous hydrogen, and one of the technologies for supplying hydrogen is through gas permeable membranes. In this study the activity and stratification of hydrogen consumption above such a membrane was investigated by use of microsensors for hydrogen and pH. A hydrogenotrophic methanogenic community that was able to consume the hydrogen flux within 0.5 mm of the membrane with specific rates of up to 30 m3 H2 m-3 day-1 developed within 3 days in fresh manure and was already established at time zero when analyzing slurry from a biogas plant. The hydrogen consumption was dependent on a simultaneous carbon dioxide supply and was inhibited when carbon dioxide depletion elevated the pH to 9.2. The activity was only partially restored when the carbon dioxide supply was resumed. Bioreactors supplied with hydrogen gas should thus be carefully monitored and either have the hydrogen supply disrupted or be supplemented with carbon dioxide when the pH rises to values about 9. PMID:27582736

  6. Temperature impacts differentially on the methanogenic food web of cellulose-supplemented peatland soil.

    PubMed

    Schmidt, Oliver; Horn, Marcus A; Kolb, Steffen; Drake, Harold L

    2015-03-01

    The impact of temperature on the largely unresolved intermediary ecosystem metabolism and associated unknown microbiota that link cellulose degradation and methane production in soils of a moderately acidic (pH 4.5) fen was investigated. Supplemental [(13) C]cellulose stimulated the accumulation of propionate, acetate and carbon dioxide as well as initial methane production in anoxic peat soil slurries at 15°C and 5°C. Accumulation of organic acids at 15°C was twice as fast as that at 5°C. 16S rRNA [(13) C]cellulose stable isotope probing identified novel unclassified Bacteria (79% identity to the next cultured relative Fibrobacter succinogenes), unclassified Bacteroidetes (89% identity to Prolixibacter bellariivorans), Porphyromonadaceae, Acidobacteriaceae and Ruminococcaceae as main anaerobic degraders of cellulose-derived carbon at both 15°C and 5°C. Holophagaceae and Spirochaetaceae were more abundant at 15°C. Clostridiaceae dominated the degradation of cellulose-derived carbon only at 5°C. Methanosarcina was the dominant methanogenic taxa at both 15°C and 5°C. Relative abundance of Methanocella increased at 15°C whereas that of Methanoregula and Methanosaeta increased at 5°C. Thaumarchaeota closely related to Nitrosotalea (presently not known to grow anaerobically) were abundant at 5°C but absent at 15°C indicating that Nitrosotalea sp. might be capable of anaerobic growth at low temperatures in peat. PMID:24813682

  7. Methanosarcina horonobensis sp. nov., a methanogenic archaeon isolated from a deep subsurface Miocene formation.

    PubMed

    Shimizu, Satoru; Upadhye, Rahul; Ishijima, Yoji; Naganuma, Takeshi

    2011-10-01

    A methanogenic organism, designated strain HB-1(T), from the domain Archaea was isolated from groundwater sampled from a subsurface Miocene formation located in Horonobe, Hokkaido, Japan. The strain grew on methanol, dimethylamine, trimethylamine, dimethylsulfide and acetate but not on monomethylamine, H(2)/CO(2), formate, 2-propanol, 2-butanol or cyclopentanol. Cells were Gram-reaction-negative, non-motile, irregular cocci that were 1.4-2.9 µm in diameter and occurred singly or in pairs. The strain grew at 20-42 °C (optimum 37 °C), at pH 6.0-7.75 (optimum pH 7.0-7.25) and in 0-0.35 M NaCl (optimum 0.1 M). The G+C content of the genomic DNA was 41.4 mol%. 16S rRNA gene sequencing revealed that the strain was a member of the genus Methanosarcina but that it clearly differed from all recognized species of this genus (93.1-97.9 % sequence similarity). The phenotypic and phylogenetic features of strain HB-1(T) indicate that it represents a novel species of the genus Methanosarcina, for which the name Methanosarcina horonobensis sp. nov. is proposed. The type strain is HB-1(T) ( = DSM 21571(T)  = JCM 15518(T)  = NBRC 102577(T)). PMID:21112985

  8. Methanosarcina subterranea sp. nov., a methanogenic archaeon isolated from a deep subsurface diatomaceous shale formation.

    PubMed

    Shimizu, Satoru; Ueno, Akio; Naganuma, Takeshi; Kaneko, Katsuhiko

    2015-04-01

    A methanogenic archaeon, strain HC-2(T), was isolated from a deep diatomaceous shale formation. The strain grew on methanol, monomethylamine, dimethylamine, trimethylamine and dimethylsulphide, but not on acetate, H2/CO2, formate, 2-propanol, 2-butanol or cyclopentanol. Cells were Gram-stain-negative, non-motile, and coccus-like, 0.9-1.4 µm in diameter, and occurred singly, in pairs, or as aggregates. The strain grew at 10-40 °C (optimum 35 °C), pH 5.9-7.4 (optimum pH 6.6-6.8) and in 0-0.6 M NaCl (optimum 0.1-0.2 M). The genomic DNA G+C content was 41.5 mol% and the 16S rRNA gene sequence was closely related to those of Methanosarcina lacustris DSM 13486(T) (99.1%) and Methanosarcina siciliae DSM 3028(T) (98.3%). Values for DNA-DNA hybridization with these strains were less than 30%. The phenotypic and phylogenetic features of HC-2(T) indicate that it represents a novel species of the genus Methanosarcina , for which the name Methanosarcina subterranea sp. nov. is proposed. The type strain is HC-2(T) ( = DSM 22503(T) = JCM 15540(T) = NBRC 102578(T)). PMID:25604340

  9. Methanogenic degradation kinetics of phenolic compounds in aquifer-derived microcosms

    USGS Publications Warehouse

    Godsy, E.M.; Goerlitz, D.F.; Grbic-Galic, D.

    1992-01-01

    In this segment of a larger multidisciplinary study of the movement and fate of creosote derived compounds in a sand-and-gravel aquifer, we present evidence that the methanogenic degradation of the major biodegradable phenolic compounds and concomitant microbial growth in batch microcosms derived from contaminated aquifer material can be described using Monod kinetics. Substrate depletion and bacterial growth curves were fitted to the Monod equations using nonlinear regression analysis. The method of Marquardt was used for the determination of parameter values that best fit the experimental data by minimizing the residual sum of squares. The Monod kinetic constants (??max, Ks, Y, and kd) that describe phenol, 2-, 3-, and 4-methylphenol degradation and concomitant microbial growth were determined under conditions that were substantially different from those previously reported for microcosms cultured from sewage sludge. The Ks values obtained in this study are approximately two orders of magnitude lower than values obtained for the anaerobic degradation of phenol in digesting sewage sludge, indicating that the aquifer microorganisms have developed enzyme systems that are adapted to low nutrient conditions. The values for kd are much less than ??max, and can be neglected in the microcosms. The extremely low Y values, approximately 3 orders of magnitude lower than for the sewage sludge derived cultures, and the very low numbers of microorganisms in the aquifer derived microcosms suggest that these organisms use some unique strategies to survive in the subsurface environment. ?? 1992 Kluwer Academic Publishers.

  10. Structural, mass and elemental analyses of storage granules in methanogenic archaeal cells

    PubMed Central

    Toso, Daniel B.; Henstra, Anne M.; Gunsalus, Robert P.; Zhou, Z. Hong

    2013-01-01

    Summary Storage granules are an important component of metabolism in many organisms spanning the bacterial, eukaryal and archaeal domains, but systematic analysis of their organization inside cells is lacking. In this study, we identify and characterize granulelike inclusion bodies in a methanogenic archaeon, Methanospirillum hungatei, an anaerobic microorganism that plays an important role in nutrient recycling in the ecosystem. Using cryo electron microscopy, we show that granules in mature M. hungatei are amorphous in structure with a uniform size. Energy dispersive X-ray spectroscopy analysis establishes that each granule is a polyphosphate body (PPB) that consists of high concentrations of phosphorous and oxygen, and increased levels of iron and magnesium. By scanning transmission electron tomography, we further estimate that the mass density within a PPB is a little less than metal titanium at room temperature and is about four times higher than that of the surrounding cytoplasm. Finally, three-dimensional cryo electron tomography reveals that PPBs are positioned off-centre in their radial locations relative to the cylindrical axis of the cell, and almost uniformly placed near cell ends. This positioning ability points to a genetic program that spatially and temporally directs the accumulation of polyphosphate into a storage granule, perhaps for energy-consuming activities, such as cell maintenance, division or motility. PMID:21854518

  11. Enhancement of anaerobic methanogenesis at a short hydraulic retention time via bioelectrochemical enrichment of hydrogenotrophic methanogens.

    PubMed

    Li, Yang; Zhang, Yaobin; Liu, Yiwen; Zhao, Zhiqiang; Zhao, Zisheng; Liu, Sitong; Zhao, Huimin; Quan, Xie

    2016-10-01

    Anaerobic digestion (AD) is an important energy strategy for converting organic waste to CH4. A major factor limiting the practical applicability of AD is the relatively long hydraulic retention time (HRT) which declines the treatment efficiency of digesters. A coupling process of anaerobic digestion and 'electromethanogenesis' was proposed to enhance anaerobic digestion at a short HRT in this study. Microorganisms analysis indicated that the electric-biological reactor enriched hydrogenotrophic methanogens in both cathodic biofilm and suspended sludge, helping achieve the high organic removal (71.0% vs 42.3% [control reactor]) and CH4 production (248.5mL/h vs 51.3mL/h), while the additional electric input was only accounted for 25.6% of the energy income from the increased CH4 production. This study demonstrated that a bioelectrochemical enhanced anaerobic reactor could improve the CH4 production and organic removal at a short HRT, providing an economically feasible scheme to treat wastewater. PMID:27394997

  12. Pressure-Enhanced Activity and Stability of a Hyperthermophilic Protease from a Deep-Sea Methanogen

    PubMed Central

    Michels, P. C.; Clark, D. S.

    1997-01-01

    We describe the properties of a hyperthermophilic, barophilic protease from Methanococcus jannaschii, an extremely thermophilic deep-sea methanogen. This enzyme is the first protease to be isolated from an organism adapted to a high-pressure-high-temperature environment. The partially purified enzyme has a molecular mass of 29 kDa and a narrow substrate specificity with strong preference for leucine at the P1 site of polypeptide substrates. Enzyme activity increased up to 116(deg)C and was measured up to 130(deg)C, one of the highest temperatures reported for the function of any enzyme. In addition, enzyme activity and thermostability increased with pressure: raising the pressure to 500 atm increased the reaction rate at 125(deg)C 3.4-fold and the thermostability 2.7-fold. Spin labeling of the active-site serine revealed that the active-site geometry of the M. jannaschii protease is not grossly different from that of several mesophilic proteases; however, the active-site structure may be relatively rigid at moderate temperatures. The barophilic and thermophilic behavior of the enzyme is consistent with the barophilic growth of M. jannaschii observed previously (J. F. Miller et al., Appl. Environ. Microbiol. 54:3039-3042, 1988). PMID:16535711

  13. Comprehensive modeling of methanogenic biofilms in fluidized bed systems: Mass transfer limitations and multisubstrate aspects

    SciTech Connect

    Buffiere, P.; Steyer, J.P.; Moletta, R.; Fonade, C.

    1995-12-20

    Anaerobic digestion--a process where organic matter is degraded into a gas mixture of methane and carbon dioxide--is particularly adapted for food industry wastewater treatment because it reduces the chemical oxygen demand (COD) of the influent and produces valuable energy (methane). A cognitive model for anaerobic digestion in fluidized bed reactors is developed. The general pathway of the process is divided into five main reactions performed by different bacterial groups. Molecular diffusion of each substrate involved in the reaction scheme is described. Effectiveness factor calculations are performed in steady state for each bacterial group taken into account in the process. The case of a single substrate removal is discussed, and optimal biofilm sizes are found. Sequential substrate removal is investigated, and different kinetic regimes are characterized. The influence of biofilm size and primary substrate removal is discussed in the case of standard concentrations in the liquid phase. This study shows that, according to the theoretical model the limiting step of the process may be different and depends in a large way on mass transfer effects. Finally, importance of biofilm size is compared for acidogenic and methanogenic steps: each reaction is found to be optimized for different biofilm thicknesses. This result may be of interest for design purposes and further dynamic modeling. Concluding remarks concerning the validation of the model are made, and a comparison to experimental data from the literature is presented.

  14. Enrichment and Characterization of a Psychrotolerant Consortium Degrading Crude Oil Alkanes Under Methanogenic Conditions.

    PubMed

    Ding, Chen; Ma, Tingting; Hu, Anyi; Dai, Lirong; He, Qiao; Cheng, Lei; Zhang, Hui

    2015-08-01

    Anaerobic alkane degradation via methanogenesis has been intensively studied under mesophilic and thermophilic conditions. While there is a paucity of information on the ability and composition of anaerobic alkane-degrading microbial communities under low temperature conditions. In this study, we investigated the ability of consortium Y15, enriched from Shengli oilfield, to degrade hydrocarbons under different temperature conditions (5-35 °C). The consortium could use hexadecane over a low temperature range (15-30 °C). No growth was detected below 10 °C and above 35 °C, indicating the presence of cold-tolerant species capable of alkane degradation. The preferential degradation of short chain n-alkanes from crude oil was observed by this consortium. The structure and dynamics of the microbial communities were examined using terminal restriction fragment length polymorphism (T-RFLP) fingerprinting and Sanger sequencing of 16S rRNA genes. The core archaeal communities were mainly composed of aceticlastic Methanosaeta spp. Syntrophaceae-related microorganisms were always detected during consecutive transfers and dominated the bacterial communities, sharing 94-96 % sequence similarity with Smithella propionica strain LYP(T). Phylogenetic analysis of Syntrophaceae-related clones in diverse methanogenic alkane-degrading cultures revealed that most of them were clustered into three sublineages. Syntrophaceae clones retrieved from this study were mainly clustered into sublineage I, which may represent psychrotolerant, syntrophic alkane degraders. These results indicate the wide geographic distribution and ecological function of syntrophic alkane degraders. PMID:25783218

  15. The presence of hydrogenotrophic methanogens in the inoculum improves methane gas production in microbial electrolysis cells

    PubMed Central

    Siegert, Michael; Li, Xiu-Fen; Yates, Matthew D.; Logan, Bruce E.

    2015-01-01

    High current densities in microbial electrolysis cells (MECs) result from the predominance of various Geobacter species on the anode, but it is not known if archaeal communities similarly converge to one specific genus. MECs were examined here on the basis of maximum methane production and current density relative to the inoculum community structure. We used anaerobic digester (AD) sludge dominated by acetoclastic Methanosaeta, and an anaerobic bog sediment where hydrogenotrophic methanogens were detected. Inoculation using solids to medium ratio of 25% (w/v) resulted in the highest methane production rates (0.27 mL mL−1 cm−2, gas volume normalized by liquid volume and cathode projected area) and highest peak current densities (0.5 mA cm−2) for the bog sample. Methane production was independent of solid to medium ratio when AD sludge was used as the inoculum. 16S rRNA gene community analysis using pyrosequencing and quantitative PCR confirmed the convergence of Archaea to Methanobacterium and Methanobrevibacter, and of Bacteria to Geobacter, despite their absence in AD sludge. Combined with other studies, these findings suggest that Archaea of the hydrogenotrophic genera Methanobacterium and Methanobrevibacter are the most important microorganisms for methane production in MECs and that their presence in the inoculum improves the performance. PMID:25642216

  16. Micro-scale H2-CO2 Dynamics in a Hydrogenotrophic Methanogenic Membrane Reactor.

    PubMed

    Garcia-Robledo, Emilio; Ottosen, Lars D M; Voigt, Niels V; Kofoed, M W; Revsbech, Niels P

    2016-01-01

    Biogas production is a key factor in a sustainable energy supply. It is possible to get biogas with very high methane content if the biogas reactors are supplied with exogenous hydrogen, and one of the technologies for supplying hydrogen is through gas permeable membranes. In this study the activity and stratification of hydrogen consumption above such a membrane was investigated by use of microsensors for hydrogen and pH. A hydrogenotrophic methanogenic community that was able to consume the hydrogen flux within 0.5 mm of the membrane with specific rates of up to 30 m(3) H2 m(-3) day(-1) developed within 3 days in fresh manure and was already established at time zero when analyzing slurry from a biogas plant. The hydrogen consumption was dependent on a simultaneous carbon dioxide supply and was inhibited when carbon dioxide depletion elevated the pH to 9.2. The activity was only partially restored when the carbon dioxide supply was resumed. Bioreactors supplied with hydrogen gas should thus be carefully monitored and either have the hydrogen supply disrupted or be supplemented with carbon dioxide when the pH rises to values about 9. PMID:27582736

  17. Recovery of Elemental Tellurium Nanoparticles by the Reduction of Tellurium Oxyanions in a Methanogenic Microbial Consortium

    PubMed Central

    Ramos-Ruiz, Adriana; Field, Jim A.; Wilkening, Jean V.; Sierra-Alvarez, Reyes

    2016-01-01

    This research focuses on the microbial recovery of elemental tellurium (Te0) from aqueous streams containing soluble tellurium oxyanions, tellurate (TeVI) and tellurite (TeIV). An anaerobic mixed microbial culture occurring in methanogenic granular sludge was able to biocatalyze the reduction of both Te oxyanions to produce Te0 nanoparticles (NPs) in sulfur-free medium. TeIV reduction was 7-fold faster than that of TeVI, such that TeIV did not accumulate to a great extent during TeVI reduction. Endogenous substrates in the granular sludge provided the electron equivalents required to reduce Te oxyanions; however, the reduction rates were modestly increased with an exogenous electron donor such as H2. The effect of four redox mediators (anthraquinone-2,6-disulfonate, hydroxocobalamin, riboflavin, and lawsone) was also tested. Riboflavin increased the rate of TeIV reduction by 11-fold and also enhanced the fraction Te recovered as extracellular Te0 NPs from 21% to 64%. Lawsone increased the rate of TeVI reduction by 5-fold and the fraction of Te recovered as extracellular material increased from 49% to 83%. The redox mediators and electron donors also impacted the morphologies and localization of Te0 NPs, suggesting that NP production can be tailored for a particular application. PMID:26735010

  18. Impact of methanogenic pre-treatment on the performance of an aerobic MBR system.

    PubMed

    Sánchez, A; Buntner, D; Garrido, J M

    2013-03-01

    The combination of anaerobic treatment with an aerobic MBR as a polishing step is an alternative to treat some industrial wastewater and/or urban wastewaters generated in warm climate countries. In this work a pilot-scale UASB reactor and an aerobic MBR as a polishing step was operated. The impact of the methanogenic stage on membrane fouling was studied. Operating fluxes of 11-18 L m(-2) h(-1) and permeabilities of 100-250 L m(-2) h(-1) bar(-1) were reported. It was demonstrated that the recirculation of aerobic biomass to the anaerobic stage provoked a release of biopolymers due to the hydrolysis of aerobic biomass in these conditions. Depending on biomass concentration in membrane chamber, the presence of biopolymers worsened membrane performance. Fouling rate was three times higher when biomass concentration decreased from 8 to 2 g L(-1), with similar concentrations of biopolymers present. Moreover, the presence of plastic support in the aerobic stage was shown to improve membrane performance, decreasing the concentrations of the studied fouling indicators. Carbohydrate fraction of soluble microbial products, biopolymer clusters (BPC) and transparent exopolymer particles (TEP) concentrations were studied as possible fouling indicators for this system. A strong correlation between both colloidal fraction of BPC (cBPC) and TEP with membrane fouling rate was observed. PMID:23245539

  19. Transformation of trinitrotoluene to triaminotoluene by mixed cultures incubated under methanogenic conditions

    SciTech Connect

    Hwang, P.; Chow, T.; Adrian, N.R.

    2000-04-01

    2,4,6-Trinitrotoluene (TNT) is an explosive widely used by the military. Although it is no longer manufactured in the US, large amounts of wastewater are generated annually from load, assembly, packing, and demilitarization operations. Granular-activated carbon adsorption is the standard technology for treating wastewater containing TNT and maintaining discharges within the limits established under the National Pollutant Discharge Elimination System. Studies evaluating biological treatment of pink water with an anaerobic fluidized-bed, granular-activated carbon bioreactor have been promising, but the fate of TNT is unknown. The authors investigated the anaerobic transformation of TNT by biofilm microorganisms obtained from a wastewater treatment plant receiving explosive manufacturing wastewater. The TNT was transformed to a mixture of 2-amino-4,6-dinitrotoluene; 4-amino-2,6-dinitrotoluene; 2,4-diamino-6-nitrotoluene; and 2,6-diamino-4-nitrotoluene before culminating in the formation of triaminotoluene (TAT). Triaminotoluene was susceptible to further degradation under anaerobic conditions, but its fate was not determined. Methane formation was inhibited but resumed after the depletion of the diaminonitrotoluene isomers. These studies demonstrate near stoichiometric formation of TAT from TNT and the transformation of 2-amino-4,6-dinitrotoluene to 2,4-diamino-6-nitrotoluene and 2,6-diamino-4-nitrotoluene by a mixed culture incubated under methanogenic conditions. This evidence indicates TAT is also a likely end-product of TNT biodegradation in the anaerobic fluidized fed bioreactor.

  20. Effect of isobutanol on toluene biodegradation in nitrate amended, sulfate amended and methanogenic enrichment microcosms.

    PubMed

    Jayamani, Indumathy; Cupples, Alison M

    2013-09-01

    Isobutanol is an alternate fuel additive that is being considered because of economic and lower emission benefits. However, future gasoline spills could result in co-contamination of isobutanol with gasoline components such as benzene, toluene, ethyl-benzene and xylene. Hence, isobutanol could affect the degradability of gasoline components thereby having an effect on contaminant plume length and half-life. In this study, the effect of isobutanol on the biodegradation of a model gasoline component (toluene) was examined in laboratory microcosms. For this, toluene and isobutanol were added to six different toluene degrading laboratory microcosms under sulfate amended, nitrate amended or methanogenic conditions. While toluene biodegradation was not greatly affected in the presence of isobutanol in five out of the six different experimental sets, toluene degradation was completely inhibited in one set of microcosms. This inhibition occurred in sulfate amended microcosms constructed with inocula from wastewater treatment plant activated sludge. Our data suggest that toluene degrading consortia are affected differently by isobutanol addition. These results indicate that, if co-contamination occurs, in some cases the in situ half-life of toluene could be significantly extended. PMID:23224907

  1. Isolation and characterization of an H/sub 2/-oxidizing thermophilic methanogen

    SciTech Connect

    Ferguson, T.J.; Mah, R.A.

    1983-01-01

    A thermophilic methanogen was isolated from enrichment cultures originally inoculated with sludge from an anaerobic kelp digester (55 degrees C). This isolate exhibited a temperature optimum of 55 to 60 degrees C and a maximum near 70 degrees C. Growth occurred throughout the pH range of 5.5 to 9.0, with optimal growth near pH 7.2. Altough 4% salt was present in the isolation medium, salt was not required for optimal growth. The thermophile utilized formate or H/sub 2/CO/sub 2/ but not acetate, methanol, or methylamines for growth and methanogenesis. Growth in complex medium was very rapid, and a minimum doubling time of 1.8 hours was recorded in media supplemented with rumen fluid. Growth in defined media required the addition of acetate and an unknown factor(s) from digester supernatant, rumen fluid, or Trypticase. Cells in liquid culture were oval to coccoid, 0.7 to 1.8 ..mu.. meters in diameter, often occurring in pairs. The cells were easily lysed upon exposure to oxygen or 0.08 mg of sodium dodecyl sulfate per ml. The isolate was sensitive to tetracycline and chloramphenicol but not penicillin G or cycloserine. The DNA base composition was 59.69 mol% guanine plus cytosine. (Refs. 34).

  2. Methanogenic biodegradation of creosote contaminants in natural and simulated ground-water ecosystems

    USGS Publications Warehouse

    Godsy, E. Michael; Goerlitz, Donald; Grbic-Galic, Dunja

    1992-01-01

    Wastes from a wood preserving plant in Pensacola, Florida have contaminated the near-surface sand-and-gravel aquifer with creosote-derived compounds and pentachlorophenol. Contamination resulted from the discharge of plant waste waters t