Selenate removal in methanogenic and sulfate-reducing upflow anaerobic sludge bed reactors

NARCIS (Netherlands)

Lenz, M.; Hullebusch, van E.D.; Hommes, G.; Corvini, P.F.X.; Lens, P.N.L.

2008-01-01

This paper evaluates the use of upflow anaerobic sludge bed (UASB) bioreactors (30 degrees C, pH = 7.0) to remove selenium oxyanions from contaminated waters (790 mu g Se L-1) under methanogenic and sulfate-reducing conditions using lactate as electron donor. One UASB reactor received sulfate at

Toxicity assessment of inorganic nanoparticles to acetoclastic and hydrogenotrophic methanogenic activity in anaerobic granular sludge.

Science.gov (United States)

Gonzalez-Estrella, Jorge; Sierra-Alvarez, Reyes; Field, James A

2013-09-15

Release of engineered nanoparticles (NPs) to municipal wastewater from industrial and residential sources could impact biological systems in wastewater treatment plants. Methanogenic inhibition can cause failure of anaerobic waste(water) treatment. This study investigated the inhibitory effect of a wide array of inorganic NPs (Ag(0), Al₂O₃, CeO₂, Cu(0), CuO, Fe(0), Fe₂O₃, Mn₂O₃, SiO₂, TiO₂, and ZnO supplied up to 1500 mgL(-1)) to acetoclastic and hydrogenotrophic methanogenic activity of anaerobic granular sludge. Of all the NPs tested, only Cu(0) and ZnO caused severe methanogenic inhibition. The 50% inhibiting concentrations determined towards acetoclastic and hydrogenotrophic methanogens were 62 and 68 mgL(-1) for Cu(0) NP; and 87 and 250 mgL(-1) for ZnO NP, respectively. CuO NPs also caused inhibition of acetoclastic methanogens. Cu(2+) and Zn(2+) salts caused similar levels of inhibition as Cu(0) and ZnO NPs based on equilibrium soluble metal concentrations measured during the assays, suggesting that the toxicity was due to the release of metal ions by NP-corrosion. A commercial dispersant, Dispex, intended to increase NP stability did not affect the inhibitory impact of the NPs. The results taken as a whole suggest that Zn- and Cu-containing NPs can release metal ions that are inhibitory for methanogenesis. Copyright © 2013 Elsevier B.V. All rights reserved.

Dosing of anaerobic granular sludge bioreactors with cobalt: Impact of cobalt retention on methanogenic activity

KAUST Repository

Fermoso, Fernando G.

2010-12-01

The effect of dosing a metal limited anaerobic sludge blanket (UASB) reactor with a metal pulse on the methanogenic activity of granular sludge has thus far not been successfully modeled. The prediction of this effect is crucial in order to optimize the strategy for metal dosage and to prevent unnecessary losses of resources. This paper describes the relation between the initial immobilization of cobalt in anaerobic granular sludge cobalt dosage into the reactor and the evolution of methanogenic activity during the subsequent weeks. An operationally defined parameter (A0· B0) was found to combine the amount of cobalt immobilized instantaneously upon the pulse (B0) and the amount of cobalt immobilized within the subsequent 24. h (A0). In contrast with the individual parameters A0 and B0, the parameter A0· B0 correlated significantly with the methanogenic activity of the sludge during the subsequent 16 or 35. days. This correlation between metal retention and activity evolution is a useful tool to implement trace metal dosing strategies for biofilm-based biotechnological processes. © 2010.

Studying gene regulation in methanogenic archaea.

Science.gov (United States)

Rother, Michael; Sattler, Christian; Stock, Tilmann

2011-01-01

Methanogenic archaea are a unique group of strictly anaerobic microorganisms characterized by their ability, and dependence, to convert simple C1 and C2 compounds to methane for growth. The major models for studying the biology of methanogens are members of the Methanococcus and Methanosarcina species. Recent development of sophisticated tools for molecular analysis and for genetic manipulation allows investigating not only their metabolism but also their cell cycle, and their interaction with the environment in great detail. One aspect of such analyses is assessment and dissection of methanoarchaeal gene regulation, for which, at present, only a handful of cases have been investigated thoroughly, partly due to the great methodological effort required. However, it becomes more and more evident that many new regulatory paradigms can be unraveled in this unique archaeal group. Here, we report both molecular and physiological/genetic methods to assess gene regulation in Methanococcus maripaludis and Methanosarcina acetivorans, which should, however, be applicable for other methanogens as well. Copyright © 2011 Elsevier Inc. All rights reserved.

The dominant acetate degradation pathway/methanogenic composition in full-scale anaerobic digesters operating under different ammonia levels

DEFF Research Database (Denmark)

Fotidis, Ioannis; Karakashev, Dimitar Borisov; Angelidaki, Irini

2014-01-01

Ammonia is a major environmental factor influencing biomethanation in full-scale anaerobic digesters. In this study, the effect of different ammonia levels on methanogenic pathways and methanogenic community composition of full-scale biogas plants was investigated. Eight full-scale digesters...... operating under different ammonia levels were sampled, and the residual biogas production was followed in fed-batch reactors. Acetate, labelled in the methyl group, was used to determine the methanogenic pathway by following the 14CH4 and 14CO2 production. Fluorescence in situ hybridisation was used...... to determine the methanogenic communities’ composition. Results obtained clearly demonstrated that syntrophic acetate oxidation coupled with hydrogenotrophic methanogenesis was the dominant pathway in all digesters with high ammonia levels (2.8–4.57 g NH4 +-N L−1), while acetoclastic methanogenic pathway...

Performance and methanogenic community of rotating disk reactor packed with polyurethane during thermophilic anaerobic digestion

International Nuclear Information System (INIS)

Yang, Yingnan; Tsukahara, Kenichiro; Sawayama, Shigeki

2007-01-01

A newly developed anaerobic rotating disk reactor (ARDR) packed with polyurethane was used in continuous mode for organic waste removal under thermophilic (55 o C) anaerobic conditions. This paper reports the effects of the rotational speed on the methanogenic performance and community in an ARDR supplied with acetic acid synthetic wastewater as the organic substrate. The best performance was obtained from the ARDR with the rotational speed (ω) of 30 rpm. The average removal of dissolved organic carbon was 98.5%, and the methane production rate was 393 ml/l-reactor/day at an organic loading rate of 2.69 g/l-reactor/day. Under these operational conditions, the reactor had a greater biomass retention capacity and better reactor performance than those at other rotational speeds (0, 5 and 60 rpm). The results of 16S rRNA phylogenetic analysis indicated that the major methanogens in the reactor belonged to the genus Methanosarcina spp. The results of real-time polymerase chain reaction (PCR) analysis suggested that the cell density of methanogenic archaea immobilized on the polyurethane foam disk could be concentrated more than 2000 times relative to those in the original thermophilic sludge. Scanning electron microphotographs showed that there were more immobilized microbes at ω of 30 rpm than 60 rpm. A rotational speed on the outer layer of the disk of 6.6 m/min could be appropriate for anaerobic digestion using the polyurethane ARDR

Fate of neptunium in an anaerobic, methanogenic microcosm

International Nuclear Information System (INIS)

Banaszak, J.E.; Webb, S.M.; Rittmann, B.E.; Gaillard, J.F.; Reed, D.T.

1999-01-01

Neptunium is found predominantly as Np(IV) in reducing environments, but as Np(V) in aerobic environments. Currently, it is not known how the interplay between biotic and abiotic processes affects Np redox speciation in the environment. To evaluate the effect of anaerobic microbial activity on the fate of Np in natural systems, Np(V) was added to a microcosm inoculated with anaerobic sediments from a metal-contaminated freshwater 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 -5 M Np did not inhibit methane production. Total Np solubility 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 microbially produced Mn(II/III) and Fe(II) may serve as electron donors for Np reduction

Dosing of anaerobic granular sludge bioreactors with cobalt: Impact of cobalt retention on methanogenic activity

KAUST Repository

Fermoso, Fernando G.; Bartacek, Jan; Manzano, Ramon; van Leeuwen, Herman P.; Lens, Piet N.L.

2010-01-01

The effect of dosing a metal limited anaerobic sludge blanket (UASB) reactor with a metal pulse on the methanogenic activity of granular sludge has thus far not been successfully modeled. The prediction of this effect is crucial in order to optimize

New approach to control the methanogenic reactor of a two-phase anaerobic digestion system.

Science.gov (United States)

von Sachs, Jürgen; Meyer, Ulrich; Rys, Paul; Feitkenhauer, Heiko

2003-03-01

A new control strategy for the methanogenic reactor of a two-phase anaerobic digestion system has been developed and successfully tested on the laboratory scale. The control strategy serves the purpose to detect inhibitory effects and to achieve good conversion. The concept is based on the idea that volatile fatty acids (VFA) can be measured in the influent of the methanogenic reactor by means of titration. Thus, information on the output (methane production) and input of the methanogenic reactor is available, and a (carbon) mass balance can be obtained. The control algorithm comprises a proportional/integral structure with the ratio of (a) the methane production rate measured online and (b) a maximum methane production rate expected (derived from the stoichiometry) as a control variable. The manipulated variable is the volumetric feed rate. Results are shown for an experiment with VFA (feed) concentration ramps and for experiments with sodium chloride as inhibitor.

Phylogenetic comparison of the methanogenic communities from an acidic, oligotrophic fen and an anaerobic digester treating municipal wastewater sludge.

Science.gov (United States)

Steinberg, Lisa M; Regan, John M

2008-11-01

Methanogens play a critical role in the decomposition of organics under anaerobic conditions. The methanogenic consortia in saturated wetland soils are often subjected to large temperature fluctuations and acidic conditions, imposing a selective pressure for psychro- and acidotolerant community members; however, methanogenic communities in engineered digesters are frequently maintained within a narrow range of mesophilic and circumneutral conditions to retain system stability. To investigate the hypothesis that these two disparate environments have distinct methanogenic communities, the methanogens in an oligotrophic acidic fen and a mesophilic anaerobic digester treating municipal wastewater sludge were characterized by creating clone libraries for the 16S rRNA and methyl coenzyme M reductase alpha subunit (mcrA) genes. A quantitative framework was developed to assess the differences between these two communities by calculating the average sequence similarity for 16S rRNA genes and mcrA within a genus and family using sequences of isolated and characterized methanogens within the approved methanogen taxonomy. The average sequence similarities for 16S rRNA genes within a genus and family were 96.0 and 93.5%, respectively, and the average sequence similarities for mcrA within a genus and family were 88.9 and 79%, respectively. The clone libraries of the bog and digester environments showed no overlap at the species level and almost no overlap at the family level. Both libraries were dominated by clones related to uncultured methanogen groups within the Methanomicrobiales, although members of the Methanosarcinales and Methanobacteriales were also found in both libraries. Diversity indices for the 16S rRNA gene library of the bog and both mcrA libraries were similar, but these indices indicated much lower diversity in the 16S digester library than in the other three libraries.

Long-term effects of operating temperature and sulphate addition on the methanogenic community structure of anaerobic hybrid reactors.

Science.gov (United States)

Pender, Seán; Toomey, Margaret; Carton, Micheál; Eardly, Dónal; Patching, John W; Colleran, Emer; O'Flaherty, Vincent

2004-02-01

The diversity, population dynamics, and activity profiles of methanogens in anaerobic granular sludges from two anaerobic hybrid reactors treating a molasses wastewater both mesophilically (37 degrees C) and thermophilically (55 degrees C) during a 1081 day trial were determined. The influent to one of the reactors was supplemented with sulphate, after an acclimation period of 112 days, to determine the effect of competition with sulphate-reducing bacteria on the methanogenic community structure. Sludge samples were removed from the reactors at intervals throughout the operational period and examined by amplified ribosomal DNA (rDNA) restriction analysis (ARDRA) and partial sequencing of 16S rRNA genes. In total, 18 operational taxonomic units (OTUs) were identified, 12 of which were sequenced. The methanogenic communities in both reactors changed during the operational period. The seed sludge and the reactor biomass sampled during mesophilic operation, both in the presence and absence of sulphate, was characterised by a predominance of Methanosaeta spp. Following temperature elevation, the dominant methanogenic sequences detected in the non-sulphate supplemented reactor were closely related to Methanocorpusculum parvum. By contrast, the dominant OTUs detected in the sulphate-supplemented reactor upon temperature increase were related to the hydrogen-utilising methanogen, Methanobacterium thermoautotrophicum. The observed methanogenic community structure in the reactors correlated with the operational performance of the reactors during the trial and with physiological measurements of the reactor biomass. Both reactors achieved chemical oxygen demand (COD) removal efficiencies of over 90% during mesophilic operation, with or without sulphate supplementation. During thermophilic operation, the presence of sulphate resulted in decreased reactor performance (effluent acetate concentrations of >3000 mg/l and biogas methane content of acetate at 55 degrees C was

Methanomethylovorans thermophila sp. nov., a thermophilic, methylotrophic methanogen form an anaerobic reactor fed with methanol

NARCIS (Netherlands)

Jiang, B.; Parshina, S.N.; Doesburg, van W.C.J.; Lomans, B.P.; Stams, A.J.M.

2005-01-01

A novel thermophilic, obligately methylotrophic, methanogenic archaeon, strain L2FAWT, was isolated from a thermophilic laboratory-scale upflow anaerobic sludge blanket reactor fed with methanol as the carbon and energy source. Cells of strain L2FAWT were non-motile, irregular cocci, 0·7¿1·5 µm in

The contribution of fermentative bacteria and methanogenic archaea to azo dye reduction by a thermophilic anaerobic consortium

NARCIS (Netherlands)

Santos, dos A.B.; Cervantes, F.J.; Madrid, de M.P.; Bok, de F.A.M.; Stams, A.J.M.; Lier, van J.B.

2006-01-01

The contribution of fermentative bacteria and methanogenic archaea to azo dye reduction by a thermophilic anaerobic consortium was studied. Additionally, the effects of different electron-donating substrates and the redox mediator riboflavin on dye reduction were assessed by using either a

Thermophilic anaerobic acetate-utilizing methanogens and their metabolism

DEFF Research Database (Denmark)

Mladenovska, Zuzana

Six strains of thermophilic anaerobic acetate-utilizing methanogens were isolated from different full-scale thermophilic biogas plants in China and Denmark. The strain isolated from the Chinese biogas plant was designated KN-6P and the isolates from the Danish full-scale biogas plants were......, utilizing the substrates acetate, methanol and methylamines but not hydrogen/carbon dioxide. Strain Methanosarcina sp. SO-2P was able to grow mixotrophically on methanol and hydrogen/carbon dioxide with methane formation from hydrogen and carbon dioxide occurring after methanol depletion. All six...... designated HG-1P, LVG-4P R1-1P, SO-2P and V-1P. The isolates were characterized morphologically and physiologically, and their immunological and phylogenetic relatedness to already known isolated strains were established. All isolated strains were identified as organisms belonging to genus Methanosarcina...

Biogas production and methanogenic archaeal community in mesophilic and thermophilic anaerobic co-digestion processes.

Science.gov (United States)

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. Copyright © 2014 Elsevier Ltd. All rights reserved.

Application of real-time PCR to determination of combined effect of antibiotics on Bacteria, Methanogenic Archaea, Archaea in anaerobic sequencing batch reactors.

Science.gov (United States)

Aydin, Sevcan; Ince, Bahar; Ince, Orhan

2015-06-01

This study evaluated the long-term effects of erythromycin-tetracycline-sulfamethoxazole (ETS) and sulfamethoxazole-tetracycline (ST) antibiotic combinations on the microbial community and examined the ways in which these antimicrobials impact the performance of anaerobic reactors. Quantitative real-time PCR was used to determine the effect that different antibiotic combinations had on the total and active Bacteria, Archae and Methanogenic Archae. Three primer sets that targeted metabolic genes encoding formylterahydrofolate synthetase, methyl-coenzyme M reductase and acetyl-coA synthetase were also used to determine the inhibition level on the mRNA expression of the homoacetogens, methanogens and specifically acetoclastic methanogens, respectively. These microorganisms play a vital role in the anaerobic degradation of organic waste and targeting these gene expressions offers operators or someone at a treatment plant the potential to control and the improve the anaerobic system. The results of the investigation revealed that acetogens have a competitive advantage over Archaea in the presence of ETS and ST combinations. Although the efficiency with which methane production takes place and the quantification of microbial populations in both the ETS and ST reactors decreased as antibiotic concentrations increased, the ETS batch reactor performed better than the ST batch reactor. According to the expression of genes results, the syntrophic interaction of acetogens and methanogens is critical to the performance of the ETS and ST reactors. Failure to maintain the stability of these microorganisms resulted in a decrease in the performance and stability of the anaerobic reactors. Copyright © 2015 Elsevier Ltd. All rights reserved.

Anaerobic biodegradability and methanogenic toxicity of key constituents in copper chemical mechanical planarization effluents of the semiconductor industry.

Science.gov (United States)

Hollingsworth, Jeremy; Sierra-Alvarez, Reyes; Zhou, Michael; Ogden, Kimberly L; Field, Jim A

2005-06-01

Copper chemical mechanical planarization (CMP) effluents can account for 30-40% of the water discharge in semiconductor manufacturing. CMP effluents contain high concentrations of soluble copper and a complex mixture of organic constituents. The aim of this study is to perform a preliminary assessment of the treatability of CMP effluents in anaerobic sulfidogenic bioreactors inoculated with anaerobic granular sludge by testing individual compounds expected in the CMP effluents. Of all the compounds tested (copper (II), benzotriazoles, polyethylene glycol (M(n) 300), polyethylene glycol (M(n) 860) monooleate, perfluoro-1-octane sulfonate, citric acid, oxalic acid and isopropanol) only copper was found to be inhibitory to methanogenic activity at the concentrations tested. Most of the organic compounds tested were biodegradable with the exception of perfluoro-1-octane sulfonate and benzotriazoles under sulfate reducing conditions and with the exception of the same compounds as well as Triton X-100 under methanogenic conditions. The susceptibility of key components in CMP effluents to anaerobic biodegradation combined with their low microbial inhibition suggest that CMP effluents should be amenable to biological treatment in sulfate reducing bioreactors.

Vitamin and Amino Acid Auxotrophy in Anaerobic Consortia Operating under Methanogenic Conditions.

Science.gov (United States)

Hubalek, Valerie; Buck, Moritz; Tan, BoonFei; Foght, Julia; Wendeberg, Annelie; Berry, David; Bertilsson, Stefan; Eiler, Alexander

2017-01-01

Syntrophy among Archaea and Bacteria facilitates the anaerobic degradation of organic compounds to CH 4 and CO 2 . Particularly during aliphatic and aromatic hydrocarbon mineralization, as in the case of crude oil reservoirs and petroleum-contaminated sediments, metabolic interactions between obligate mutualistic microbial partners are of central importance. Using micromanipulation combined with shotgun metagenomic approaches, we describe the genomes of complex consortia within short-chain alkane-degrading cultures operating under methanogenic conditions. Metabolic reconstruction revealed that only a small fraction of genes in the metagenome-assembled genomes encode the capacity for fermentation of alkanes facilitated by energy conservation linked to H 2 metabolism. Instead, the presence of inferred lifestyles based on scavenging anabolic products and intermediate fermentation products derived from detrital biomass was a common feature. Additionally, inferred auxotrophy for vitamins and amino acids suggests that the hydrocarbon-degrading microbial assemblages are structured and maintained by multiple interactions beyond the canonical H 2 -producing and syntrophic alkane degrader-methanogen partnership. Compared to previous work, our report points to a higher order of complexity in microbial consortia engaged in anaerobic hydrocarbon transformation. IMPORTANCE Microbial interactions between Archaea and Bacteria mediate many important chemical transformations in the biosphere from degrading abundant polymers to synthesis of toxic compounds. Two of the most pressing issues in microbial interactions are how consortia are established and how we can modulate these microbial communities to express desirable functions. Here, we propose that public goods (i.e., metabolites of high energy demand in biosynthesis) facilitate energy conservation for life under energy-limited conditions and determine the assembly and function of the consortia. Our report suggests that an

Conductive properties of methanogenic biofilms.

Science.gov (United States)

Li, Cheng; Lesnik, Keaton Larson; Liu, Hong

2018-02-01

Extracellular electron transfer between syntrophic partners needs to be efficiently maintained in methanogenic environments. Direct extracellular electron transfer via electrical current is an alternative to indirect hydrogen transfer but requires construction of conductive extracellular structures. Conductive mechanisms and relationship between conductivity and the community composition in mixed-species methanogenic biofilms are not well understood. The present study investigated conductive behaviors of methanogenic biofilms and examined the correlation between biofilm conductivity and community composition between different anaerobic biofilms enriched from the same inoculum. Highest conductivity observed in methanogenic biofilms was 71.8±4.0μS/cm. Peak-manner response of conductivity upon changes over a range of electrochemical potentials suggests that electron transfer in methanogenic biofilms occurs through redox driven super-exchange. The strong correlation observed between biofilm conductivity and Geobacter spp. in the metabolically diverse anaerobic communities suggests that the efficiency of DEET may provide pressure for microbial communities to select for species that can produce electrical conduits. Copyright © 2017 Elsevier B.V. All rights reserved.

Anaerobic bacterium that degrades fatty acids in syntrophic association with methanogens

Energy Technology Data Exchange (ETDEWEB)

McInerney, M J [Univ. of Illinois, Urbana; Bryant, M P; Pfennig, N

1979-01-01

A new species of anaerobic bacterium that degrades the even-numbered carbon fatty acids, butyrate, caproate and caprylate, to acetate and H/sub 2/ and the odd-numbered carbon fatty acids, valerate and heptanoate, to acetate, propionate and H/sub 2/ was obtained in coculture with either an H/sub 2/-utilizing methanogen or H/sub 2/-utilizing desulfovibrio. The organism could be grown only in syntrophic association with the H/sub 2/-utilizer and no other energy sources or combination of electron donor and acceptors were utilized. It was a Gram-negative helical rod with 2 to 8 flagella, about 20 nm in diameter, inserted in a linear fashion about 130 nm or more apart along the concave side of the cell. It grew with a generation time of 84 h in co-culture with Methanospirillum hungatii and was present in numbers of at least 4.5 x 10/sup -6/ per g of anaerobic digest or sludge.

A mathematical model for the interactive behavior of sulfate-reducing bacteria and methanogens during anaerobic digestion.

Science.gov (United States)

Ahammad, S Ziauddin; Gomes, James; Sreekrishnan, T R

2011-09-01

Anaerobic degradation of waste involves different classes of microorganisms, and there are different types of interactions among them for substrates, terminal electron acceptors, and so on. A mathematical model is developed based on the mass balance of different substrates, products, and microbes present in the system to study the interaction between methanogens and sulfate-reducing bacteria (SRB). The performance of major microbial consortia present in the system, such as propionate-utilizing acetogens, butyrate-utilizing acetogens, acetoclastic methanogens, hydrogen-utilizing methanogens, and SRB were considered and analyzed in the model. Different substrates consumed and products formed during the process also were considered in the model. The experimental observations and model predictions showed very good prediction capabilities of the model. Model prediction was validated statistically. It was observed that the model-predicted values matched the experimental data very closely, with an average error of 3.9%.

Methanogenic archaea

International Nuclear Information System (INIS)

Joblin, K.N.

2005-01-01

This chapter outlines procedures for enumerating, isolating, culturing and storing methanogens from ruminal digesta. The methanogens, a large and diverse group of Archaea, have unique features that separate them from the bacteria and the eukaryotes. They are the only recognized ruminal microbes belonging to the Archaea and are an integral part of the rumen microbial ecosystem. By scavenging hydrogen gas, methanogens play a key ecological role in keeping the partial pressure of hydrogen low so that fermentation can proceed efficiently. Although about 70 methanogenic species belonging to 2 1 genera have been identified from anaerobic environments, and a range of different methanogens co-exist in the rumen, to date only seven ruminal species have been isolated and purified. The population densities of methanogens in the rumen appear to be influenced by diet, and in particular by the fibre content of the diet. Sheep and cattle fed diets rich in concentrates contained 10 7 -10 8 and 10 8 - 10 9 ruminal methanogens/g, respectively, whereas sheep and dairy cows grazing pasture contained 10 9 -10 10 ruminal methanogens/g (G.N. Jarvis and K.N. Joblin, unpublished data). With careful application, methanogen population densities can readily be determined using culture methods. These appear to be similar to the population densities determined by culture-independent methods (P. Evans and K.N. Joblin, unpublished data)

Quantification of methanogenic biomass by enzyme-linked immunosorbent assay and by analysis of specific methanogenic cofactors

Energy Technology Data Exchange (ETDEWEB)

Gorris, L G.M.; Kemp, H A; Archer, D B

1987-01-01

The reliability and accuracy with which enzyme-linked immunosorbent assay (ELISA) and an assay of methanogenic cofactors detect and quantify methanogenic species were investigated. Both assays required standardization with laboratory cultures of methanogenic bacteria and were applied to mixtures of pure cultures and samples from anaerobic digesters. ELISA was shown to be a simple method for detecting and quantifying individual methanogenic species. The range of species which can be assayed is limited by the range of antisera available but, potentially, ELISA can be applied to all methanogens. Although the cofactor assay is not species-specific it can distinguish hydrogenotrophic and acetotrophic methanogens and is quantitative.

Immobilization patterns and dynamics of acetate-utilizing methanogens in sterile granular sludge from upflow anaerobic sludge blanket (UASB) reactors

DEFF Research Database (Denmark)

Schmidt, Jens Ejbye; Ahring, Birgitte Kiær

1999-01-01

Sterile granular sludge was inoculated with either Methanosarcina mazeii S-6, Methanosaeta concilii GP-6, or both species in acetate-fea upflow anaerobic sludge blanket (UASB) reactors to investigate the immobilization patterns and dynamics of aceticlastic methanogens in granular sludge. After......, but where the acetate concentration was low this strain was immobilized on support material as single cells or small clumps, The data clearly show that the two aceticlastic methanogens immobilize differently in UASB systems, depending on the conditions found throughout the UASB reactor....

Immobilization patterns and dynamics of acetate-utilizing methanogens immobilized in sterile granular sludge in upflow anaerobic sludge blanket reactors

DEFF Research Database (Denmark)

Schmidt, Jens Ejbye; Ahring, Birgitte Kiær

1999-01-01

Sterile granular sludge was inoculated with either Methanosarcina mazeii S-6, Methanosaeta concilii GP-6, or both species in acetate-fea upflow anaerobic sludge blanket (UASB) reactors to investigate the immobilization patterns and dynamics of aceticlastic methanogens in granular sludge. After......, but where the acetate concentration was low this strain was immobilized on support material as single cells or small clumps, The data clearly show that the two aceticlastic methanogens immobilize differently in UASB systems, depending on the conditions found throughout the UASB reactor....

Growth of a Strictly Anaerobic Bacterium on Furfural (2-Furaldehyde)

OpenAIRE

Brune, Gerhard; Schoberth, Siegfried M.; Sahm, Hermann

1983-01-01

A strictly anaerobic bacterium was isolated from a continuous fermentor culture which converted the organic constituents of sulfite evaporator condensate to methane and carbon dioxide. Furfural is one of the major components of this condensate. This furfural isolate could degrade furfural as the sole source of carbon and energy in a defined mineral-vitamin-sulfate medium. Acetic acid was the major fermentation product. This organism could also use ethanol, lactate, pyruvate, or fumarate and c...

The influence of substrate transport limitation on porosity and methanogenic activity of anaerobic sludge granules

Energy Technology Data Exchange (ETDEWEB)

Alphenaar, P.A. (Agricultural Univ., Wageningen (Netherlands). Dept. of Environmental Technology); Perez, M.C. (Agricultural Univ., Wageningen (Netherlands). Dept. of Environmental Technology); Lettinga, G. (Agricultural Univ., Wageningen (Netherlands). Dept. of Environmental Technology)

1993-05-01

The relationship between porosity, diameter and methanogenic activity of anaerobic granules has been investigated. Experiments with different granular sludges revealed that substrate transport limitations increase with the diameter of the granules. As a consequence, autolysis can occur in the core of the granule, producing hollow granules. The porosity measurements revealed that the hollow centre is not available for substrate transport. Possibly as an effect of bacterial lysis, the porosity decreases in the more interior layers of the granules. This results in a inactive inner part of the large granules, which is not involved in the treatment process; the specific methanogenic activity decreases with granule size. No marked difference in substrate affinity is observed between granules of different sizes, which probably indicates that for large granules only the exterior is biological active. (orig.)

A strict anaerobic extreme thermophilic hydrogen-producing culture enriched from digested household waste

DEFF Research Database (Denmark)

Karakashev, Dimitar Borisov; Kotay, Shireen Meher; Trably, Eric

2009-01-01

The aim of this study was to enrich, characterize and identify strict anaerobic extreme thermophilic hydrogen (H-2) producers from digested household solid wastes. A strict anaerobic extreme thermophilic H-2 producing bacterial culture was enriched from a lab-scale digester treating household...... wastes at 70 degrees C. The enriched mixed culture consisted of two rod-shaped bacterial members growing at an optimal temperature of 80 degrees C and an optimal pH 8.1. The culture was able to utilize glucose, galactose, mannose, xylose, arabinose, maltose, sucrose, pyruvate and glycerol as carbon...... sources. Growth on glucose produced acetate, H-2 and carbon dioxide. Maximal H-2 production rate on glucose was 1.1 mmol l(-1) h(-1) with a maximum H-2 yield of 1.9 mole H-2 per mole glucose. 16S ribosomal DNA clone library analyses showed that the culture members were phylogenetically affiliated...

Morphology and Phylogeny of a New Species of Anaerobic Ciliate, Trimyema finlayi n. sp., with Endosymbiotic Methanogens.

Science.gov (United States)

Lewis, William H; Sendra, Kacper M; Embley, T Martin; Esteban, Genoveva F

2018-01-01

Many anaerobic ciliated protozoa contain organelles of mitochondrial ancestry called hydrogenosomes. These organelles generate molecular hydrogen that is consumed by methanogenic Archaea, living in endosymbiosis within many of these ciliates. Here we describe a new species of anaerobic ciliate, Trimyema finlayi n. sp., by using silver impregnation and microscopy to conduct a detailed morphometric analysis. Comparisons with previously published morphological data for this species, as well as the closely related species, Trimyema compressum , demonstrated that despite them being similar, both the mean cell size and the mean number of somatic kineties are lower for T. finlayi than for T. compressum , which suggests that they are distinct species. This was also supported by analysis of the 18S rRNA genes from these ciliates, the sequences of which are 97.5% identical (6 substitutions, 1479 compared bases), and in phylogenetic analyses these sequences grouped with other 18S rRNA genes sequenced from previous isolates of the same respective species. Together these data provide strong evidence that T. finlayi is a novel species of Trimyema , within the class Plagiopylea. Various microscopic techniques demonstrated that T. finlayi n. sp. contains polymorphic endosymbiotic methanogens, and analysis of the endosymbionts' 16S rRNA gene showed that they belong to the genus Methanocorpusculum , which was confirmed using fluorescence in situ hybridization with specific probes. Despite the degree of similarity and close relationship between these ciliates, T. compressum contains endosymbiotic methanogens from a different genus, Methanobrevibacter . In phylogenetic analyses of 16S rRNA genes, the Methanocorpusculum endosymbiont of T. finlayi n. sp. grouped with sequences from Methanomicrobia, including the endosymbiont of an earlier isolate of the same species, ' Trimyema sp.,' which was sampled approximately 22 years earlier, at a distant (∼400 km) geographical location

Morphology and Phylogeny of a New Species of Anaerobic Ciliate, Trimyema finlayi n. sp., with Endosymbiotic Methanogens

Directory of Open Access Journals (Sweden)

William H. Lewis

2018-02-01

Full Text Available Many anaerobic ciliated protozoa contain organelles of mitochondrial ancestry called hydrogenosomes. These organelles generate molecular hydrogen that is consumed by methanogenic Archaea, living in endosymbiosis within many of these ciliates. Here we describe a new species of anaerobic ciliate, Trimyema finlayi n. sp., by using silver impregnation and microscopy to conduct a detailed morphometric analysis. Comparisons with previously published morphological data for this species, as well as the closely related species, Trimyema compressum, demonstrated that despite them being similar, both the mean cell size and the mean number of somatic kineties are lower for T. finlayi than for T. compressum, which suggests that they are distinct species. This was also supported by analysis of the 18S rRNA genes from these ciliates, the sequences of which are 97.5% identical (6 substitutions, 1479 compared bases, and in phylogenetic analyses these sequences grouped with other 18S rRNA genes sequenced from previous isolates of the same respective species. Together these data provide strong evidence that T. finlayi is a novel species of Trimyema, within the class Plagiopylea. Various microscopic techniques demonstrated that T. finlayi n. sp. contains polymorphic endosymbiotic methanogens, and analysis of the endosymbionts’ 16S rRNA gene showed that they belong to the genus Methanocorpusculum, which was confirmed using fluorescence in situ hybridization with specific probes. Despite the degree of similarity and close relationship between these ciliates, T. compressum contains endosymbiotic methanogens from a different genus, Methanobrevibacter. In phylogenetic analyses of 16S rRNA genes, the Methanocorpusculum endosymbiont of T. finlayi n. sp. grouped with sequences from Methanomicrobia, including the endosymbiont of an earlier isolate of the same species, ‘Trimyema sp.,’ which was sampled approximately 22 years earlier, at a distant (∼400 km

Anaerobic biodegradation of fluoranthene under methanogenic conditions in presence of surface-active compounds

DEFF Research Database (Denmark)

Fuchedzhieva, Nadezhda; Karakashev, Dimitar Borisov; Angelidaki, Irini

2008-01-01

for the study were linear alkyl benzene sulphonates (LAS) and rhamnolipid-biosurfactant complex from Pseudomonas sp. PS-17. Biodegradation of fluoranthene was monitored by GC/MS for a period up to 12th day. No change in the fluoranthene concentration was registered after 7th day. The presence of LAS enhanced...... biodegradation was most likely as a result of the increased fluoranthene solubility. The results indicate that LAS can be considered as a promising agent for facilitation of the process of anaerobic polycyclic aromatic hydrocarbons (PAH) biodegradation under methanogenic conditions....

Characterization of the methanogen community in a household anaerobic digester fed with swine manure in China.

Science.gov (United States)

Qin, Huibin; Lang, Huihua; Yang, Hongjiang

2013-09-01

Household anaerobic digesters have been installed across rural China for biogas production, but information on methanogen community structure in these small biogas units is sparsely available. By creating clone libraries for 16S rRNA and methyl coenzyme M reductase alpha subunit (mcrA) genes, we investigated the methanogenic consortia in a household biogas digester treating swine manure. Operational taxonomic units (OTUs) were defined by comparative sequence analysis, seven OTUs were identified in the 16S rRNA gene library, and ten OTUs were identified in the mcrA gene library. Both libraries were dominated by clones highly related to the type strain Methanocorpusculum labreanum Z, 64.0 % for 16S rRNA gene clones and 64.3 % for mcrA gene clones. Additionally, gas chromatography assays showed that formic acid was 84.54 % of the total volatile fatty acids and methane was 57.20 % of the biogas composition. Our results may help further isolation and characterization of methanogenic starter strains for industrial biogas production.

A new methanogen "Methanobrevibacter massiliense" isolated in a case of severe periodontitis.

Science.gov (United States)

Huynh, Hong T T; Pignoly, Marion; Drancourt, Michel; Aboudharam, Gérard

2017-12-01

A few methanogens have been previously recovered from periodontitis lesions, yet their repertoire may not be completed. We recovered a previously unreported methanogen species in this situation. A 64-year-old Caucasian woman was diagnosed with chronic, severe generalized periodontitis. In the presence of negative controls, an 18-month culture of periodontal pockets in anaerobe Hungate tube yielded "Methanobrevibacter massiliense" and Pyramidobacter piscolens. This case report provides evidence of the symbiotic strategy deployed by the methanogens and the anaerobes, and reports the first culture of a new methanogen, "M. massiliense".

INFLUENCE OF METRONIDAZOLE, CO, CO2, AND METHANOGENS ON THE FERMENTATIVE METABOLISM OF THE ANAEROBIC FUNGUS NEOCALLIMASTIX SP STRAIN L2

NARCIS (Netherlands)

MARVINSIKKEMA, FD; REES, E; KRAAK, MN; GOTTSCHAL, JC; PRINS, RA

The effects of metronidazole, CO, methanogens, and CO, on the fermentation of glucose by the anaerobic fungus Neocallimastix sp. strain L2 were investigated. Both metronidazole and CO caused a shift in the fermentation products from predominantly H-2, acetate, and formate to lactate as the major

My Lifelong Passion for Biochemistry and Anaerobic Microorganisms.

Science.gov (United States)

Thauer, Rudolf Kurt

2015-01-01

Early parental influence led me first to medical school, but after developing a passion for biochemistry and sensing the need for a deeper foundation, I changed to chemistry. During breaks between semesters, I worked in various biochemistry labs to acquire a feeling for the different areas of investigation. The scientific puzzle that fascinated me most was the metabolism of the anaerobic bacterium Clostridium kluyveri, which I took on in 1965 in Karl Decker's lab in Freiburg, Germany. I quickly realized that little was known about the biochemistry of strict anaerobes such as clostridia, methanogens, acetogens, and sulfate-reducing bacteria and that these were ideal model organisms to study fundamental questions of energy conservation, CO2 fixation, and the evolution of metabolic pathways. My passion for anaerobes was born then and is unabated even after 50 years of study.

Immobilization Patterns and Dynamics of Acetate-Utilizing Methanogens Immobilized in Sterile Granular Sludge in Upflow Anaerobic Sludge Blanket Reactors

Science.gov (United States)

Schmidt, Jens Ejbye; Ahring, Birgitte Kjær

1999-01-01

Sterile granular sludge was inoculated with either Methanosarcina mazeii S-6, Methanosaeta concilii GP-6, or both species in acetate-fed upflow anaerobic sludge blanket (UASB) reactors to investigate the immobilization patterns and dynamics of aceticlastic methanogens in granular sludge. After several months of reactor operation, the methanogens were immobilized, either separately or together. The fastest immobilization was observed in the reactor containing M. mazeii S-6. The highest effluent concentration of acetate was observed in the reactor with only M. mazeii S-6 immobilized, while the lowest effluent concentration of acetate was observed in the reactor where both types of methanogens were immobilized together. No changes were observed in the kinetic parameters (Ks and μmax) of immobilized M. concilii GP-6 or M. mazeii S-6 compared with suspended cultures, indicating that immobilization does not affect the growth kinetics of these methanogens. An enzyme-linked immunosorbent assay using polyclonal antibodies against either M. concilii GP-6 or M. mazeii S-6 showed significant variations in the two methanogenic populations in the different reactors. Polyclonal antibodies were further used to study the spatial distribution of the two methanogens. M. concilii GP-6 was immobilized only on existing support material without any specific pattern. M. mazeii S-6, however, showed a different immobilization pattern: large clumps were formed when the concentration of acetate was high, but where the acetate concentration was low this strain was immobilized on support material as single cells or small clumps. The data clearly show that the two aceticlastic methanogens immobilize differently in UASB systems, depending on the conditions found throughout the UASB reactor. PMID:10049862

Anaerobic Digestion of Sugarcane Vinasse Through a Methanogenic UASB Reactor Followed by a Packed Bed Reactor.

Science.gov (United States)

Cabrera-Díaz, A; Pereda-Reyes, I; Oliva-Merencio, D; Lebrero, R; Zaiat, M

2017-12-01

The anaerobic treatment of raw vinasse in a combined system consisting in two methanogenic reactors, up-flow anaerobic sludge blanket (UASB) + anaerobic packed bed reactors (APBR), was evaluated. The organic loading rate (OLR) was varied, and the best condition for the combined system was 12.5 kg COD m -3 day -1 with averages of 0.289 m 3 CH 4  kg COD r -1 for the UASB reactor and 4.4 kg COD m -3 day -1 with 0.207 m 3 CH 4  kg COD r -1 for APBR. The OLR played a major role in the emission of H 2 S conducting to relatively stable quality of biogas emitted from the APBR, with H 2 S concentrations <10 mg L -1 . The importance of the sulphate to COD ratio was demonstrated as a result of the low biogas quality recorded at the lowest ratio. It was possible to develop a proper anaerobic digestion of raw vinasse through the combined system with COD removal efficiency of 86.7% and higher CH 4 and a lower H 2 S content in biogas.

Adaptation of Methanogenic Inocula to Anaerobic Digestion of Maize Silage

Directory of Open Access Journals (Sweden)

Martyna Wojcieszak

2017-09-01

Full Text Available A well-balanced microbial consortium is crucial for efficient biogas production. In turn, one of a major factor that influence on the structure of anaerobic digestion (AD consortium is a source of microorganisms which are used as an inoculum. This study evaluated the influence of inoculum sources (with various origin on adaptation of a biogas community and the efficiency of the biomethanization of maize silage. As initial inocula for AD of maize silage the samples from: (i an agricultural biogas plant (ABP which utilizes maize silage as a main substrate, (ii cattle slurry (CS, which contain elevated levels of lignocelluloses materials, and (iii raw sewage sludge (RSS with low content of plant origin materials were used. The adaptation of methanogenic consortia was monitored during a series of passages, and the functionality of the adapted consortia was verified through start-up operation of AD in two-stage reactors. During the first stages of the adaptation phase, methanogenic consortia occurred very slowly, and only after several passages did the microbial community adapts to allow production of biogas with high methane content. The ABP consortium revealed highest biogas production in the adaptation and in the start-up process. The biodiversity dynamics monitored during adaptation and start-up process showed that community profile changed in a similar direction in three studied consortia. Native communities were very distinct to each other, while at the end of the Phase II of the start-up process microbial diversity profile was similar in all consortia. All adopted bacterial communities were dominated by representatives of Porphyromonadaceae, Rikenellaceae, Ruminococcaceae, and Synergistaceae. A shift from low acetate-preferring acetoclastic Methanosaetaceae (ABP and RSS and/or hydrogenotrophic Archaea, e.g., Methanomicrobiaceae (CS prevailing in the inoculum samples to larger populations of high acetate-preferring acetoclastic

Anaerobic biodegradation of cellulosic material: Batch experiments and modelling based on isotopic data and focusing on aceticlastic and non-aceticlastic methanogenesis

International Nuclear Information System (INIS)

Qu, X.; Vavilin, V.A.; Mazeas, L.; Lemunier, M.; Duquennoi, C.; He, P.-J.; Bouchez, T.

2009-01-01

Utilizing stable carbon isotope data to account for aceticlastic and non-aceticlastic pathways of methane generation, a model was created to describe laboratory batch anaerobic decomposition of cellulosic materials (office paper and cardboard). The total organic and inorganic carbon concentrations, methane production volume, and methane and CO 2 partial pressure values were used for the model calibration and validation. According to the fluorescent in situ hybridization observations, three groups of methanogens including strictly hydrogenotrophic methanogens, strictly aceticlastic methanogens (Methanosaeta sp.) and Methanosarcina sp., consuming both acetate and H 2 /H 2 CO 3 as well as acetate-oxidizing syntrophs, were considered. It was shown that temporary inhibition of aceticlastic methanogens by non-ionized volatile fatty acids or acidic pH was responsible for two-step methane production from office paper at 35 o C where during the first and second steps methane was generated mostly from H 2 /H 2 CO 3 and acetate, respectively. Water saturated and unsaturated cases were tested. According to the model, at the intermediate moisture (150%), much lower methane production occurred because of full-time inhibition of aceticlastic methanogens. At the lowest moisture, methane production was very low because most likely hydrolysis was seriously inhibited. Simulations showed that during cardboard and office paper biodegradation at 55 o C, non-aceticlastic syntrophic oxidation by acetate-oxidizing syntrophs and hydrogenotrophic methanogens were the dominant methanogenic pathways.

Effect of methanogenic substrates on anaerobic oxidation of methane and sulfate reduction by an anaerobic methanotrophic enrichment.

KAUST Repository

Meulepas, Roel J W

2010-05-06

Anaerobic oxidation of methane (AOM) coupled to sulfate reduction (SR) is assumed to be a syntrophic process, in which methanotrophic archaea produce an interspecies electron carrier (IEC), which is subsequently utilized by sulfate-reducing bacteria. In this paper, six methanogenic substrates are tested as candidate-IECs by assessing their effect on AOM and SR by an anaerobic methanotrophic enrichment. The presence of acetate, formate or hydrogen enhanced SR, but did not inhibit AOM, nor did these substrates trigger methanogenesis. Carbon monoxide also enhanced SR but slightly inhibited AOM. Methanol did not enhance SR nor did it inhibit AOM, and methanethiol inhibited both SR and AOM completely. Subsequently, it was calculated at which candidate-IEC concentrations no more Gibbs free energy can be conserved from their production from methane at the applied conditions. These concentrations were at least 1,000 times lower can the final candidate-IEC concentration in the bulk liquid. Therefore, the tested candidate-IECs could not have been produced from methane during the incubations. Hence, acetate, formate, methanol, carbon monoxide, and hydrogen can be excluded as sole IEC in AOM coupled to SR. Methanethiol did inhibit AOM and can therefore not be excluded as IEC by this study.

Integrated biogas upgrading and hydrogen utilization in an anaerobic reactor containing enriched hydrogenotrophic methanogenic culture

DEFF Research Database (Denmark)

Luo, Gang; Angelidaki, Irini

2012-01-01

Biogas produced by anaerobic digestion, is mainly used in a gas motor for heat and electricity production. However, after removal of CO2, biogas can be upgraded to natural gas quality, giving more utilization possibilities, such as utilization as autogas, or distant utilization by using...... the existing natural gas grid. The current study presents a new biological method for biogas upgrading in a separate biogas reactor, containing enriched hydrogenotrophic methanogens and fed with biogas and hydrogen. Both mesophilic- and thermophilic anaerobic cultures were enriched to convert CO2 to CH4...... by addition of H2. Enrichment at thermophilic temperature (55°C) resulted in CO2 and H2 bioconversion rate of 320 mL CH4/(gVSS h), which was more than 60% higher than that under mesophilic temperature (37°C). Different dominant species were found at mesophilic- and thermophilic-enriched cultures, as revealed...

Biostimulation of anaerobic BTEX biodegradation under fermentative methanogenic conditions at source-zone groundwater contaminated with a biodiesel blend (B20).

Science.gov (United States)

Ramos, Débora Toledo; da Silva, Márcio Luis Busi; Chiaranda, Helen Simone; Alvarez, Pedro J J; Corseuil, Henry Xavier

2013-06-01

Field experiments were conducted to assess the potential for anaerobic biostimulation to enhance BTEX biodegradation under fermentative methanogenic conditions in groundwater impacted by a biodiesel blend (B20, consisting of 20 % v/v biodiesel and 80 % v/v diesel). B20 (100 L) was released at each of two plots through an area of 1 m(2) that was excavated down to the water table, 1.6 m below ground surface. One release was biostimulated with ammonium acetate, which was added weekly through injection wells near the source zone over 15 months. The other release was not biostimulated and served as a baseline control simulating natural attenuation. Ammonium acetate addition stimulated the development of strongly anaerobic conditions, as indicated by near-saturation methane concentrations. BTEX removal began within 8 months in the biostimulated source zone, but not in the natural attenuation control, where BTEX concentrations were still increasing (due to source dissolution) 2 years after the release. Phylogenetic analysis using quantitative PCR indicated an increase in concentration and relative abundance of Archaea (Crenarchaeota and Euryarchaeota), Geobacteraceae (Geobacter and Pelobacter spp.) and sulfate-reducing bacteria (Desulfovibrio, Desulfomicrobium, Desulfuromusa, and Desulfuromonas) in the biostimulated plot relative to the control. Apparently, biostimulation fortuitously enhanced the growth of putative anaerobic BTEX degraders and associated commensal microorganisms that consume acetate and H2, and enhance the thermodynamic feasibility of BTEX fermentation. This is the first field study to suggest that anaerobic-methanogenic biostimulation could enhance source zone bioremediation of groundwater aquifers impacted by biodiesel blends.

The anaerobic digestion process

Energy Technology Data Exchange (ETDEWEB)

Rivard, C.J. [National Renewable Energy Lab., Golden, CO (United States); Boone, D.R. [Oregon Graduate Inst., Portland, OR (United States)

1996-01-01

The microbial process of converting organic matter into methane and carbon dioxide is so complex that anaerobic digesters have long been treated as {open_quotes}black boxes.{close_quotes} Research into this process during the past few decades has gradually unraveled this complexity, but many questions remain. The major biochemical reactions for forming methane by methanogens are largely understood, and evolutionary studies indicate that these microbes are as different from bacteria as they are from plants and animals. In anaerobic digesters, methanogens are at the terminus of a metabolic web, in which the reactions of myriads of other microbes produce a very limited range of compounds - mainly acetate, hydrogen, and formate - on which the methanogens grow and from which they form methane. {open_quotes}Interspecies hydrogen-transfer{close_quotes} and {open_quotes}interspecies formate-transfer{close_quotes} are major mechanisms by which methanogens obtain their substrates and by which volatile fatty acids are degraded. Present understanding of these reactions and other complex interactions among the bacteria involved in anaerobic digestion is only now to the point where anaerobic digesters need no longer be treated as black boxes.

Effect of cryopreservation and lyophilization on viability and growth of strict anaerobic human gut microbes.

Science.gov (United States)

Bircher, Lea; Geirnaert, Annelies; Hammes, Frederik; Lacroix, Christophe; Schwab, Clarissa

2018-04-17

Strict anaerobic gut microbes have been suggested as 'next-generation probiotics' for treating several intestinal disorders. The development of preservation techniques is of major importance for therapeutic application. This study investigated cryopreservation (-80°C) and lyophilization survival and storage stability (4°C for 3 months) of the strict anaerobic gut microbes Bacteroides thetaiotaomicron, Faecalibacterium prausnitzii, Roseburia intestinalis, Anaerostipes caccae, Eubacterium hallii and Blautia obeum. To improve preservation survival, protectants sucrose and inulin (both 5% w/v) were added for lyophilization and were also combined with glycerol (15% v/v) for cryopreservation. Bacterial fitness, evaluated by maximum growth rate and lag phase, viability and membrane integrity were determined using a standardized growth assay and by flow cytometry as markers for preservation resistance. Lyophilization was more detrimental to viability and fitness than cryopreservation, but led to better storage stability. Adding sucrose and inulin enhanced viability and the proportion of intact cells during lyophilization of all strains. Viability of protectant-free B. thetaiotaomicron, A. caccae and F. prausnitzii was above 50% after cryopreservation and storage and increased to above 80% if protectants were present. The addition of glycerol, sucrose and inulin strongly enhanced the viability of B. obeum, E. hallii and R. intestinalis from 0.03-2% in protectant-free cultures to 11-37%. This is the first study that quantitatively compared the effect of cryopreservation and lyophilization and the addition of selected protectants on viability and fitness of six strict anaerobic gut microbes. Our results suggest that efficiency of protectants is process- and species-specific. © 2018 The Authors. Microbial Biotechnology published by John Wiley & Sons Ltd and Society for Applied Microbiology.

Growth of a Strictly Anaerobic Bacterium on Furfural (2-Furaldehyde)

Science.gov (United States)

Brune, Gerhard; Schoberth, Siegfried M.; Sahm, Hermann

1983-01-01

A strictly anaerobic bacterium was isolated from a continuous fermentor culture which converted the organic constituents of sulfite evaporator condensate to methane and carbon dioxide. Furfural is one of the major components of this condensate. This furfural isolate could degrade furfural as the sole source of carbon and energy in a defined mineral-vitamin-sulfate medium. Acetic acid was the major fermentation product. This organism could also use ethanol, lactate, pyruvate, or fumarate and contained cytochrome c3 and desulfoviridin. Except for furfural degradation, the characteristics of the furfural isolate were remarkably similar to those of the sulfate reducer Desulfovibrio gigas. The furfural isolate has been tentatively identified as Desulfovibrio sp. strain F-1. Images PMID:16346423

Multiple Syntrophic Interactions in a Terephthalate-Degrading Methanogenic Consortium

Energy Technology Data Exchange (ETDEWEB)

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.

Methanogenic community development in anaerobic granular bioreactors treating trichloroethylene (TCE)-contaminated wastewater at 37 °C and 15 °C.

Science.gov (United States)

Siggins, Alma; Enright, Anne-Marie; O'Flaherty, Vincent

2011-04-01

Four expanded granular sludge bed (EGSB) bioreactors were seeded with a mesophilically-grown granular sludge and operated in duplicate for mesophilic (37 °C; R1 & R2) and low- (15°; R3 & R4) temperature treatment of a synthetic volatile fatty acid (VFA) based wastewater (3 kg COD m(-3) d(-1)) with one of each pair (R1 & R3) supplemented with increasing concentrations of trichloroethylene (TCE; 10, 20, 40, 60 mg l(-1)) and one acting as a control. Bioreactor performance was evaluated by % COD removal efficiency and % biogas methane (CH(4)) content. Quantitative Polymerase Chain Reaction (qPCR) was used to investigate the methanogenic community composition and dynamics in the bioreactors during the trial, while specific methanogenic activity (SMA) and toxicity assays were utilized to investigate the activity and TCE/dichloroethylene (DCE) toxicity thresholds of key trophic groups, respectively. At both 37 °C and 15 °C, TCE levels of 60 mg l(-1) resulted in the decline of % COD removal efficiencies to 29% (Day 235) and 37% (Day 238), respectively, and in % biogas CH(4) to 54% (Day 235) and 5% (Day 238), respectively. Despite the inhibitory effect of TCE on the anaerobic digestion process, the main drivers influencing methanogenic community development, as determined by qPCR and Non-metric multidimensional scaling analysis, were (i) wastewater composition and (ii) operating temperature. At the apical TCE concentration both SMA and qPCR of methanogenic archaea suggested that acetoclastic methanogens were somewhat inhibited by the presence of TCE and/or its degradation derivatives, while competition by dechlorinating organisms may have limited the availability of H(2) for hydrogenotrophic methanogenesis. In addition, there appeared to be an inverse correlation between SMA levels and TCE tolerance, a finding that was supported by the analysis of the inhibitory effect of TCE on two additional biomass sources. The results indicate that low-temperature anaerobic

Fermentative Degradation of Polyethylene Glycol by a Strictly Anaerobic, Gram-Negative, Nonsporeforming Bacterium, Pelobacter venetianus sp. nov

OpenAIRE

1983-01-01

The synthetic polyether polyethylene glycol (PEG) with a molecular weight of 20,000 was anaerobically degraded in enrichment cultures inoculated with mud of limnic and marine origins. Three strains (Gra PEG 1, Gra PEG 2, and Ko PEG 2) of rod-shaped, gram-negative, nonsporeforming, strictly anaerobic bacteria were isolated in mineral medium with PEG as the sole source of carbon and energy. All strains degraded dimers, oligomers, and polymers of PEG up to a molecular weight of 20,000 completely...

Hydrogen or formate: Alternative key players in methanogenic degradation.

Science.gov (United States)

Schink, Bernhard; Montag, Dominik; Keller, Anja; Müller, Nicolai

2017-06-01

Hydrogen and formate are important electron carriers in methanogenic degradation in anoxic environments such as sediments, sewage sludge digestors and biogas reactors. Especially in the terminal steps of methanogenesis, they determine the energy budgets of secondary (syntrophically) fermenting bacteria and their methanogenic partners. The literature provides considerable data on hydrogen pool sizes in such habitats, but little data exist for formate concentrations due to technical difficulties in formate determination at low concentration. Recent evidence from biochemical and molecular biological studies indicates that several secondary fermenters can use both hydrogen and formate for electron release, and may do so even simultaneously. Numerous strictly anaerobic bacteria contain enzymes which equilibrate hydrogen and formate pools to energetically equal values, and recent measurements in sewage digestors and biogas reactors indicate that - beyond occasional fluctuations - the pool sizes of hydrogen and formate are indeed energetically nearly equivalent. Nonetheless, a thermophilic archaeon from a submarine hydrothermal vent, Thermococcus onnurineus, can obtain ATP from the conversion of formate to hydrogen plus bicarbonate at 80°C, indicating that at least in this extreme environment the pools of formate and hydrogen are likely to be sufficiently different to support such an unusual type of energy conservation. © 2017 Society for Applied Microbiology and John Wiley & Sons Ltd.

Microbiological aspects of granular methanogenic sludge

NARCIS (Netherlands)

Dolfing, J.

1987-01-01

The settling characteristics of anaerobic sludge are enhanced by the formation of microbial conglomerates. Various types of conglomerates having different structures, were distinguished in the present study, viz. granules, pellets and flocs (chapter 1). Granular methanogenic sludge, often

Potential for direct interspecies electron transfer in methanogenic wastewater digester aggregates

DEFF Research Database (Denmark)

Morita, Masahiko; Malvankar, Nikhil S; Franks, Ashley E

2011-01-01

Mechanisms for electron transfer within microbial aggregates derived from an upflow anaerobic sludge blanket reactor converting brewery waste to methane were investigated in order to better understand the function of methanogenic consortia. The aggregates were electrically conductive, with conduc...... for electron exchange in some methanogenic systems....

ELISA techniques for the determination of methanogenic bacteria

Energy Technology Data Exchange (ETDEWEB)

Bryniok, D; Troesch, W [Fraunhofer-Institut fuer Grenzflaechen- und Bioverfahrenstechnik (IGB), Stuttgart (Germany, F.R.)

1989-12-01

Easy-to-handle enzyme-linked immunosorbent assay (ELISA) techniques have been developed suitable for quantitative species-specific determination of very low numbers of methanogens in complex bacterial populations. The amount and the distribution of different species of methanogens in anaerobic digestors is a reflection of the functional status of the degradation process; this can be recognized with these tests and hence may be used for process control. (orig.).

Application of Methanobrevibacter acididurans in anaerobic digestion.

Science.gov (United States)

Savant, D V; Ranade, D R

2004-01-01

To operate anaerobic digesters successfully under acidic conditions, hydrogen utilizing methanogens which can grow efficiently at low pH and tolerate high volatile fatty acids (VFA) are desirable. An acid tolerant hydrogenotrophic methanogen viz. Methanobrevibacter acididurans isolated from slurry of an anaerobic digester running on alcohol distillery wastewater has been described earlier by this lab. This organism could grow optimally at pH 6.0. In the experiments reported herein, M. acididurans showed better methanogenesis under acidic conditions with high VFA, particularly acetate, than Methanobacterium bryantii, a common hydrogenotrophic inhabitant of anaerobic digesters. Addition of M. acididurans culture to digesting slurry of acidogenic as well as methanogenic digesters running on distillery wastewater showed increase in methane production and decrease in accumulation of volatile fatty acids. The results proved the feasibility of application of M. acididurans in anaerobic digesters.

Syntrophorhabdus aromaticivorans gen. nov., sp. nov., the first cultured anaerobe capable of degrading phenol to acetate in obligate syntrophic associations with a hydrogenotrophic methanogen.

Science.gov (United States)

Qiu, Yan-Ling; Hanada, Satoshi; Ohashi, Akiyoshi; Harada, Hideki; Kamagata, Yoichi; Sekiguchi, Yuji

2008-04-01

Phenol degradation under methanogenic conditions has long been studied, but the anaerobes responsible for the degradation reaction are still largely unknown. An anaerobe, designated strain UI(T), was isolated in a pure syntrophic culture. This isolate is the first tangible, obligately anaerobic, syntrophic substrate-degrading organism capable of oxidizing phenol in association with an H(2)-scavenging methanogen partner. Besides phenol, it could metabolize p-cresol, 4-hydroxybenzoate, isophthalate, and benzoate. During the degradation of phenol, a small amount of 4-hydroxybenzoate (a maximum of 4 microM) and benzoate (a maximum of 11 microM) were formed as transient intermediates. When 4-hydroxybenzoate was used as the substrate, phenol (maximum, 20 microM) and benzoate (maximum, 92 microM) were detected as intermediates, which were then further degraded to acetate and methane by the coculture. No substrates were found to support the fermentative growth of strain UI(T) in pure culture, although 88 different substrates were tested for growth. 16S rRNA gene sequence analysis indicated that strain UI(T) belongs to an uncultured clone cluster (group TA) at the family (or order) level in the class Deltaproteobacteria. Syntrophorhabdus aromaticivorans gen. nov., sp. nov., is proposed for strain UI(T), and the novel family Syntrophorhabdaceae fam. nov. is described. Peripheral 16S rRNA gene sequences in the databases indicated that the proposed new family Syntrophorhabdaceae is largely represented by abundant bacteria within anaerobic ecosystems mainly decomposing aromatic compounds.

Physiologically anaerobic microorganisms of the deep subsurface

International Nuclear Information System (INIS)

Stevens, S.E. Jr.; Chung, K.T.

1993-10-01

Anaerobic bacteria were isolated from deep subsurface sediment samples taken at study sites in Idaho (INEL) and Washington (HR) by culturing on dilute and concentrated medium. Morphologically distinct colonies were purified, and their responses to 21 selected physiological tests were determined. Although the number of isolates was small (18 INEL, 27 HR) some general patterns could be determined. Most strains could utilize all the carbon sources, however the glycerol and melizitose utilization was positive for 50% or less of the HR isolates. Catalase activity (27.78% at INEL, 74.07% at HR) and tryptophan metabolism (11.12% at INEL, 40.74% at HR) were significantly different between the two study sites. MPN and viable counts indicate that sediments near the water table yield the greatest numbers of anaerobes. Deeper sediments also appear to be more selective with the greatest number of viable counts on low-nutrient mediums. Likewise, only strictly obligate anaerobes were found in the deepest sediment samples. Selective media indicated the presence of methanogens, acetogens, and sulfate reducers at only the HR site

Methanogenic degradation of toilet-paper cellulose upon sewage treatment in an anaerobic membrane bioreactor at room temperature.

Science.gov (United States)

Chen, Rong; Nie, Yulun; Kato, Hiroyuki; Wu, Jiang; Utashiro, Tetsuya; Lu, Jianbo; Yue, Shangchao; Jiang, Hongyu; Zhang, Lu; Li, Yu-You

2017-03-01

Toilet-paper cellulose with rich but refractory carbon sources, are the main insoluble COD fractions in sewage. An anaerobic membrane bioreactor (AnMBR) was configured for sewage treatment at room temperature and its performance on methanogenic degradation of toilet paper was highlighted. The results showed, high organic removal (95%), high methane conversion (90%) and low sludge yield (0.08gVSS/gCOD) were achieved in the AnMBR. Toilet-paper cellulose was fully biodegraded without accumulation in the mixed liquor and membrane cake layer. Bioconversion efficiency of toilet paper approached 100% under a high organic loading rate (OLR) of 2.02gCOD/L/d and it could provide around 26% of total methane generation at most of OLRs. Long sludge retention time and co-digestion of insoluble/soluble COD fractions achieving mutualism of functional microorganisms, contributed to biodegradation of toilet-paper cellulose. Therefore the AnMBR successfully implemented simultaneously methanogenic bioconversion of toilet-paper cellulose and soluble COD in sewage at room temperature. Copyright © 2016 Elsevier Ltd. All rights reserved.

Probing the redox metabolism in the strictly anaerobic, extremely thermophilic, hydrogen-producing Caldicellulosiruptor saccharolyticus using amperometry

DEFF Research Database (Denmark)

Kostesha, Natalie; Willquist, Karin; Emnéus, Jenny

2011-01-01

Changes in the redox metabolism in the anaerobic, extremely thermophilic, hydrogen-forming bacterium Caldicellulosiruptor saccharolyticus were probed for the first time in vivo using mediated amperometry with ferricyanide as a thermotolerant external mediator. Clear differences in the intracellul...... in the intracellular electron flow and to probe redox enzyme properties of a strictly anaerobic thermophile in vivo.......Changes in the redox metabolism in the anaerobic, extremely thermophilic, hydrogen-forming bacterium Caldicellulosiruptor saccharolyticus were probed for the first time in vivo using mediated amperometry with ferricyanide as a thermotolerant external mediator. Clear differences in the intracellular...... the NADH-dependent lactate dehydrogenase, upon which more NADH was directed to membrane-associated enzymes for ferricyanide reduction, leading to a higher electrochemical signal. The method is noninvasive and the results presented here demonstrate that this method can be used to accurately detect changes...

Microbial ecology of methanogenic crude oil biodegradation; from microbial consortia to heavy oil

Energy Technology Data Exchange (ETDEWEB)

Head, Ian M.; Maguire, Michael J.; Sherry, Angela; Grant, Russell; Gray, Neil D.; Aitken, Carolyn M.; Martin Jones, D.; Oldenburg, Thomas B.P.; Larter, Stephen R. [Petroleum Research Group, Geosciences, University of Calgary (Canada)

2011-07-01

This paper presents the microbial ecology of methanogenic crude oil biodegradation. Biodegraded petroleum reservoirs are one of the most dramatic indications of the deep biosphere. It is estimated that heavy oil and oil sands will account for a considerable amount of energy production in the future. Carbon, a major resource for deep subsurface microorganisms, and energy are contained in large quantities in petroleum reservoirs. The aerobic to anaerobic paradigm shift is explained. A key process for in-situ oil biodegradation in petroleum reservoirs is methanogenesis. New paradigms for in-reservoir crude oil biodegradation are discussed. Variations in anaerobic degradation of crude oil hydrocarbons are also discussed. A graph shows the different patterns of crude oil biodegradation under sulfate-reducing and methanogenic conditions. Alternative anaerobic alkane activation mechanisms are also shown. From the study, it can be concluded that methanogenic crude oil degradation is of global importance and led to the establishment of the world's enormous heavy oil deposits.

Different cultivation methods to acclimatise ammonia-tolerant methanogenic consortia.

Science.gov (United States)

Tian, Hailin; Fotidis, Ioannis A; Mancini, Enrico; Angelidaki, Irini

2017-05-01

Bioaugmentation with ammonia tolerant-methanogenic consortia was proposed as a solution to overcome ammonia inhibition during anaerobic digestion process recently. However, appropriate technology to generate ammonia tolerant methanogenic consortia is still lacking. In this study, three basic reactors (i.e. batch, fed-batch and continuous stirred-tank reactors (CSTR)) operated at mesophilic (37°C) and thermophilic (55°C) conditions were assessed, based on methane production efficiency, incubation time, TAN/FAN (total ammonium nitrogen/free ammonia nitrogen) levels and maximum methanogenic activity. Overall, fed-batch cultivation was clearly the most efficient method compared to batch and CSTR. Specifically, by saving incubation time up to 150%, fed-batch reactors were acclimatised to nearly 2-fold higher FAN levels with a 37%-153% methanogenic activity improvement, compared to batch method. Meanwhile, CSTR reactors were inhibited at lower ammonia levels. Finally, specific methanogenic activity test showed that hydrogenotrophic methanogens were more active than aceticlastic methanogens in all FAN levels above 540mgNH 3 -NL -1 . Copyright © 2017 Elsevier Ltd. All rights reserved.

Quantifying electron fluxes in methanogenic microbial communities

NARCIS (Netherlands)

Junicke, H.

2015-01-01

Anaerobic digestion is a widely applied process in which close interactions between different microbial groups result in the formation of renewable energy in the form of biogas. Nevertheless, the regulatory mechanisms of the electron transfer between acetogenic bacteria and methanogenic archaea in

Anaerobe Tolerance to Oxygen and the Potentials of Anaerobic and Aerobic Cocultures for Wastewater Treatment

Directory of Open Access Journals (Sweden)

M.T. Kato

1997-12-01

Full Text Available The anaerobic treatment processes are considered to be well-established methods for the elimination of easily biodegradable organic matter from wastewaters. Some difficulties concerning certain wastewaters are related to the possible presence of dissolved oxygen. The common belief is that anaerobes are oxygen intolerant. Therefore, the common practice is to use sequencing anaerobic and aerobic steps in separate tanks. Enhanced treatment by polishing off the residual biodegradable oxygen demand from effluents of anaerobic reactors, or the biodegradation of recalcitrant wastewater pollutants, usually requires sequenced anaerobic and aerobic bacteria activities. However, the combined activity of both bacteria can also be obtained in a single reactor. Previous experiments with either pure or mixed cultures showed that anaerobes can tolerate oxygen to a certain extent. The oxygen toxicity to methanogens in anaerobic sludges was quantified in batch experiments, as well as in anaerobic reactors. The results showed that methanogens have a high tolerance to oxygen. In practice, it was confirmed that dissolved oxygen does not constitute any detrimental effect on reactor treatment performance. This means that the coexistence of anaerobic and aerobic bacteria in one single reactor is feasible and increases the potentials of new applications in wastewater treatment

Evidence of syntrophic acetate oxidation by Spirochaetes during anaerobic methane production.

Science.gov (United States)

Lee, Sang-Hoon; Park, Jeong-Hoon; Kim, Sang-Hyoun; Yu, Byung Jo; Yoon, Jeong-Jun; Park, Hee-Deung

2015-08-01

To search for evidence of syntrophic acetate oxidation by cluster II Spirochaetes with hydrogenotrophic methanogens, batch reactors seeded with five different anaerobic sludge samples supplemented with acetate as the sole carbon source were operated anaerobically. The changes in abundance of the cluster II Spirochaetes, two groups of acetoclastic methanogens (Methanosaetaceae and Methanosarcinaceae), and two groups of hydrogenotrophic methanogens (Methanomicrobiales and Methanobacteriales) in the reactors were assessed using qPCR targeting the 16S rRNA genes of each group. Increase in the cluster II Spirochaetes (9.0±0.4-fold) was positively correlated with increase in hydrogenotrophic methanogens, especially Methanomicrobiales (5.6±1.0-fold), but not with acetoclastic methanogens. In addition, the activity of the cluster II Spirochaetes decreased (4.6±0.1-fold) in response to high hydrogen partial pressure, but their activity was restored after consumption of hydrogen by the hydrogenotrophic methanogens. These results strongly suggest that the cluster II Spirochaetes are involved in syntrophic acetate oxidation in anaerobic digesters. Copyright © 2015 Elsevier Ltd. All rights reserved.

A new combination of substrates: biogas production and diversity of the methanogenic microorganisms

Directory of Open Access Journals (Sweden)

Kushkevych Ivan

2018-04-01

Full Text Available Agriculture, food industry, and manufacturing are just some of the areas where anaerobic technology can be used. Currently, anaerobic technologies are mainly used for wastewater treatment, solid waste treatment, or for the production of electrical and thermal energy from energy crops processing. However, a clear trend is towards more intensive use of this technology in biomass and biodegradable waste processing and hydrogen or biomethane production. An enormous number of anaerobic digesters are operating worldwide but there is very little information about the effect of different substrate combinations on the methanogens community. This is due to the fact that each of the anaerobic digesters has its own unique microbial community. For the most effective management of anaerobic processes it would be important to know the composition of a consortium of anaerobic microorganisms present in anaerobic digesters processing different input combinations of raw material. This paper characterizes the effect of the input raw materials on the diversity of the methanogen community. Two predominant microorganisms in anaerobic digesters were found to be 99% identity by the sequences of the 16S rRNA gene to the Methanoculleus and Thermogymnomonas genera deposited in GenBank.

Ammonia effect on hydrogenotrophic methanogens and syntrophic acetate oxidizing bacteria

DEFF Research Database (Denmark)

Wang, Han; Fotidis, Ioannis; Angelidaki, Irini

Substrates that contain high ammonia levels can cause inhibition on anaerobic digestion process and unstable biogas production. The aim of the current study was to assess the effects of different ammonia levels on pure strains of (syntrophic acetate oxidizing) SAO bacteria and hydrogenotrophic...... methanogens. Two pure strains of hydrogenotrophic methanogens (i.e: Methanoculleus bourgensis and Methanoculleus thermophiles) and two pure strains of SAO bacteria (i.e: Tepidanaerobacter acetatoxydans and Thermacetogenium phaeum) were inoculated under four different ammonia (0.26, 3, 5 and 7g NH4+-N......, the total incubation periods of hydrogenotrophic methanogens were significantly shorter compared to the SAO bacteria incubation periods. Thus, it seems that hydrogenotrophic methanogens could be equally, if not more, tolerant to high ammonia levels compared to SAO bacteria....

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

Science.gov (United States)

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. Copyright 2010 Elsevier Inc. All rights reserved.

Methanogenic activity tests by Infrared Tunable Diode Laser Absorption Spectroscopy.

Science.gov (United States)

Martinez-Cruz, Karla; Sepulveda-Jauregui, Armando; Escobar-Orozco, Nayeli; Thalasso, Frederic

2012-10-01

Methanogenic activity (MA) tests are commonly carried out to estimate the capability of anaerobic biomass to treat effluents, to evaluate anaerobic activity in bioreactors or natural ecosystems, or to quantify inhibitory effects on methanogenic activity. These activity tests are usually based on the measurement of the volume of biogas produced by volumetric, pressure increase or gas chromatography (GC) methods. In this study, we present an alternative method for non-invasive measurement of methane produced during activity tests in closed vials, based on Infrared Tunable Diode Laser Absorption Spectroscopy (MA-TDLAS). This new method was tested during model acetoclastic and hydrogenotrophic methanogenic activity tests and was compared to a more traditional method based on gas chromatography. From the results obtained, the CH(4) detection limit of the method was estimated to 60 ppm and the minimum measurable methane production rate was estimated to 1.09(.)10(-3) mg l(-1) h(-1), which is below CH(4) production rate usually reported in both anaerobic reactors and natural ecosystems. Additionally to sensitivity, the method has several potential interests compared to more traditional methods among which short measurements time allowing the measurement of a large number of MA test vials, non-invasive measurements avoiding leakage or external interferences and similar cost to GC based methods. It is concluded that MA-TDLAS is a promising method that could be of interest not only in the field of anaerobic digestion but also, in the field of environmental ecology where CH(4) production rates are usually very low. Copyright © 2012 Elsevier B.V. All rights reserved.

Kinetic and thermodynamic control of butyrate conversion in non-defined methanogenic communities.

Science.gov (United States)

Junicke, H; van Loosdrecht, M C M; Kleerebezem, R

2016-01-01

Many anaerobic conversions proceed close to thermodynamic equilibrium and the microbial groups involved need to share their low energy budget to survive at the thermodynamic boundary of life. This study aimed to investigate the kinetic and thermodynamic control mechanisms of the electron transfer during syntrophic butyrate conversion in non-defined methanogenic communities. Despite the rather low energy content of butyrate, results demonstrate unequal energy sharing between the butyrate-utilizing species (17 %), the hydrogenotrophic methanogens (9-10 %), and the acetoclastic methanogens (73-74 %). As a key finding, the energy disproportion resulted in different growth strategies of the syntrophic partners. Compared to the butyrate-utilizing partner, the hydrogenotrophic methanogens compensated their lower biomass yield per mole of electrons transferred with a 2-fold higher biomass-specific electron transfer rate. Apart from these thermodynamic control mechanisms, experiments revealed a ten times lower hydrogen inhibition constant on butyrate conversion than proposed by the Anaerobic Digestion Model No. 1, suggesting a much stronger inhibitory effect of hydrogen on anaerobic butyrate conversion. At hydrogen partial pressures exceeding 40 Pa and at bicarbonate limited conditions, a shift from methanogenesis to reduced product formation was observed which indicates an important role of the hydrogen partial pressure in redirecting electron fluxes towards reduced products such as butanol. The findings of this study demonstrate that a careful consideration of thermodynamics and kinetics is required to advance our current understanding of flux regulation in energy-limited syntrophic ecosystems.

Growth of methylaminotrophic, acetotrophic and hydrogenotrophic methanogenic bacteria on artificial supports.

Science.gov (United States)

Urrutia, H; Vidal, R; Baeza, M; Reyes, J E; Aspe, E

1997-06-01

The efficiency of organic matter degradation in attached biomass reactors depends on the suitable selection of artificial support for the retention of bacterial communities. We have studied the growth on glass and clay beads of methylaminotrophic, acetotrophic and hydrogenotrophic methanogenic bacterial communities isolated from anaerobic reactors. Bacterial counts were performed by the standard MPN technique. Experiments were performed in 50 ml vials for 12 days at 35 degrees C. Increase in the counts of methylaminotrophic and hydrogenotrophic methanogens occurred on both glass and clay beads. The latter support material also stimulated the growth rate of methylaminotrophic methanogens.

Intra-Genomic Heterogeneity in 16S rRNA Genes in Strictly Anaerobic Clinical Isolates from Periodontal Abscesses.

Science.gov (United States)

Chen, Jiazhen; Miao, Xinyu; Xu, Meng; He, Junlin; Xie, Yi; Wu, Xingwen; Chen, Gang; Yu, Liying; Zhang, Wenhong

2015-01-01

Members of the genera Prevotella, Veillonella and Fusobacterium are the predominant culturable obligate anaerobic bacteria isolated from periodontal abscesses. When determining the cumulative number of clinical anaerobic isolates from periodontal abscesses, ambiguous or overlapping signals were frequently encountered in 16S rRNA gene sequencing chromatograms, resulting in ambiguous identifications. With the exception of the genus Veillonella, the high intra-chromosomal heterogeneity of rrs genes has not been reported. The 16S rRNA genes of 138 clinical, strictly anaerobic isolates and one reference strain were directly sequenced, and the chromatograms were carefully examined. Gene cloning was performed for 22 typical isolates with doublet sequencing signals for the 16S rRNA genes, and four copies of the rrs-ITS genes of 9 Prevotella intermedia isolates were separately amplified by PCR, sequenced and compared. Five conserved housekeeping genes, hsp60, recA, dnaJ, gyrB1 and rpoB from 89 clinical isolates of Prevotella were also amplified by PCR and sequenced for identification and phylogenetic analysis along with 18 Prevotella reference strains. Heterogeneity of 16S rRNA genes was apparent in clinical, strictly anaerobic oral bacteria, particularly in the genera Prevotella and Veillonella. One hundred out of 138 anaerobic strains (72%) had intragenomic nucleotide polymorphisms (SNPs) in multiple locations, and 13 strains (9.4%) had intragenomic insertions or deletions in the 16S rRNA gene. In the genera Prevotella and Veillonella, 75% (67/89) and 100% (19/19) of the strains had SNPs in the 16S rRNA gene, respectively. Gene cloning and separate amplifications of four copies of the rrs-ITS genes confirmed that 2 to 4 heterogeneous 16S rRNA copies existed. Sequence alignment of five housekeeping genes revealed that intra-species nucleotide similarities were very high in the genera Prevotella, ranging from 94.3-100%. However, the inter-species similarities were

Microbiology and performance of a methanogenic biofilm reactor during the start-up period.

Science.gov (United States)

Cresson, R; Dabert, P; Bernet, N

2009-03-01

To understand the interactions between anaerobic biofilm development and process performances during the start-up period of methanogenic biofilm reactor. Two methanogenic inverse turbulent bed reactors have been started and monitored for 81 days. Biofilm development (adhesion, growth, population dynamic) and characteristics (biodiversity, structure) were investigated using molecular tools (PCR-SSCP, FISH-CSLM). Identification of the dominant populations, in relation to process performances and to the present knowledge of their metabolic activities, was used to propose a global scheme of the degradation routes involved. The inoculum, which determines the microbial species present in the biofilm influences bioreactor performances during the start-up period. FISH observations revealed a homogeneous distribution of the Archaea and bacterial populations inside the biofilm. This study points out the link between biodiversity, functional stability and methanogenic process performances during start-up of anaerobic biofilm reactor. It shows that inoculum and substrate composition greatly influence biodiversity, physiology and structure of the biofilm. The combination of molecular techniques associated to a biochemical engineering approach is useful to get relevant information on the microbiology of a methanogenic growing biofilm, in relation with the start-up of the process.

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

Science.gov (United States)

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.

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

Directory of Open Access Journals (Sweden)

Shailendra Yadav

2015-01-01

Full Text Available 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.

A new methanogen “Methanobrevibacter massiliense” isolated in a case of severe periodontitis

OpenAIRE

Huynh, Hong T. T.; Pignoly, Marion; Drancourt, Michel; Aboudharam, Gérard

2017-01-01

Background A few methanogens have been previously recovered from periodontitis lesions, yet their repertoire may not be completed. We recovered a previously unreported methanogen species in this situation. Case presentation A 64-year-old Caucasian woman was diagnosed with chronic, severe generalized periodontitis. In the presence of negative controls, an 18-month culture of periodontal pockets in anaerobe Hungate tube yielded “Methanobrevibacter massiliense” and Pyramidobacter piscolens. Conc...

Effects of high salinity wastewater on methanogenic sludge bed systems

NARCIS (Netherlands)

Ismail, S.; Gonzalez-Contreras, P.A.; Jeison, D.A.; Lier, van J.B.

2008-01-01

The attainable loading potentials of anaerobic sludge bed systems are strongly dependent on the growth of granular biomass with a particular wastewater. Experiments were conducted to determine the effects of high salinity wastewater on the biological and physical properties of methanogenic sludge.

HIGH-RATE ANAEROBIC TREATMENT OF ALCOHOLIC WASTEWATERS

Directory of Open Access Journals (Sweden)

Florencio L.

1997-01-01

Full Text Available Modern high-rate anaerobic wastewater treatment processes are rapidly becoming popular for industrial wastewater treatment. However, until recently stable process conditions could not be guaranteed for alcoholic wastewaters containing higher concentrations of methanol. Although methanol can be directly converted into methane by methanogens, under specific conditions it can also be converted into acetate and butyrate by acetogens. The accumulation of volatile fatty acids can lead to reactor instability in a weakly buffered reactor. Since this process was insufficiently understood, the application of high-rate anaerobic reactors was highly questionable. This research investigated the environmental factors that are of importance in the predominance of methylotrophic methanogens over acetogens in a natural mixed culture during anaerobic wastewater treatment in upflow anaerobic sludge bed reactors. Technological and microbiological aspects were investigated. Additionally, the route by which methanol is converted into methane is also presented

Molecular genetic studies on obligate anaerobic bacteria

International Nuclear Information System (INIS)

Woods, D.R.

1982-01-01

Molecular genetic studies on obligate anaerobic bacteria have lagged behind similar studies in aerobes. However, the current interest in biotechnology, the involvement of anaerobes in disease and the emergence of antibioticresistant strains have focused attention on the genetics of anaerobes. This article reviews molecular genetic studies in Bacteroides spp., Clostridium spp. and methanogens. Certain genetic systems in some anaerobes differ from those in aerobes and illustrate the genetic diversity among bacteria

Development of a PCR assay based on the 16S-23S rDNA internal transcribed spacer for identification of strictly anaerobic bacterium Zymophilus

Czech Academy of Sciences Publication Activity Database

Felsberg, Jürgen; Jelínková, Markéta; Kubizniaková, P.; Matoulková, D.

2015-01-01

Roč. 33, June (2015), s. 85-89 ISSN 1075-9964 Institutional support: RVO:61388971 Keywords : Brewing microbiology * Strictly anaerobic bacteria * Yeast contamination Subject RIV: GM - Food Processing Impact factor: 2.424, year: 2015

Mineralization of LCFA associated with anaerobic sludge: Kinetics, enhancement of methanogenic activity, and effect of VFA.

Science.gov (United States)

Pereira, M A; Sousa, D Z; Mota, M; Alves, M M

2004-11-20

Long-chain fatty acids (LCFA) associated with anaerobic sludge by mechanisms of precipitation, adsorption, or entrapment can be biodegraded to methane. The mineralization kinetics of biomass-associated LCFA were established according to an inhibition model based on Haldane's enzymatic inhibition kinetics. A value around 1,000 mg COD-LCFA..g VSS(-1) was obtained for the optimal specific LCFA content that allowed the maximal mineralization rate. For sludge with specific LCFA contents of 2,838 +/- 63 and 4,571 +/- 257 mg COD-LCFA..g VSS(-1), the specific methanogenic activities in the presence of acetate, butyrate, and H(2)/CO(2) were significantly enhanced after the mineralization of the biomass-associated LCFA. For sludge with a specific LCFA content near the optimal value defined by the kinetic model, the effect of adding VFA to the medium was studied during the mineralization of the biomass-associated LCFA. Different patterns were obtained for each individual substrate. Acetate and butyrate were preferentially consumed by the consortium, but in the case of propionate no evidence of a sequential consumption pattern could be withdrawn. It was concluded that LCFA do not exert a bactericidal neither a permanent toxic effect toward the anaerobic consortia. A discussion is addressed to the relative roles of a reversible inhibitory effect and a transport limitation effect imposed by the LCFA surrounding the cells. (c) 2004 Wiley Periodicals, Inc

Involvement of formate as an interspecies electron carrier in a syntrophic acetate-oxidizing anaerobic microorganism in coculture with methanogens.

Science.gov (United States)

Hattori, S; Luo, H; Shoun, H; Kamagata, Y

2001-01-01

To determine whether formate is involved in interspecies electron transfer between substrate-oxidizing bacteria and hydrogenotrophic microorganisms under anaerobic conditions, a syntrophic acetate-oxidizing bacterium Thermacetogenium phaeum strain PB was cocultured either with a formate /H2-utilizing methanogen strain TM (designated as PB/TM coculture), or an H2-utilizing methanogen strain deltaH (designated as PB/deltaH coculture). Acetate oxidation and subsequent methanogenesis in PB/TM coculture were found to be significantly faster than in PB/deltaH coculture. Formate dehydrogenase and hydrogenase were both detected in strains PB and TM. H2 partial pressures in the PB/TM coculture were kept lower (20 to 40 Pa) than those of the PB/deltaH coculture (40 to 60 Pa) during the exponential growth phase. Formate was also detected in both PB/TM and PB/deltaH cocultures, and the concentration of formate was maintained at a lower level in the PB/TM coculture (5 to 9 microM) than in the PB/deltaH coculture. Thermodynamic calculations revealed that the concentrations of both H2 and formate severely affect the syntrophic oxidation of acetate. These results strongly indicate that not only H2 but also formate may be involved in interspecies electron transfer.

Methane fermentation process as anaerobic digestion of biomass ...

African Journals Online (AJOL)

Anaerobic decomposition of organic compounds is conducted in close cooperation of specialized bacteria of different types, including mostly hydrolyzing, digestive, acetogenic, homoacetogenic, sulfate-reducing (VI) and methanogenic bacteria. A great interest in the anaerobic digestion process results mainly from its ...

Molecular Biology and Physiology of Methanogenic Archaebacteria

Science.gov (United States)

1989-06-27

anaerobic food chains, the methanogens contribute to the mineralization of large amounts of organic matter. The end product of their metabolism...of radiolabelled substrate to product [8; Worrell and Nagle, in preparation]. Strain RT103, a formate auxotroph was isolated from the kanamycin...methylmercaptopurine riboside 0. 16 Bacteriocidald 8-aza-2, 6-diaminopurine 0.0011 6-thioguanine 0.0004 8-azaguanine 0.0004 6- mercaptopurine 0 8

Anaerobic digestion of cheese whey using an upflow anaerobic sludge blanket reactor: Pt. 3; Sludge and substrate profiles

Energy Technology Data Exchange (ETDEWEB)

Yan, J.Q.; Lo, K.V.; Liao, P.H. (British Columbia Univ., Vancouver (CA). Dept. of Bio-Resource Engineering)

1990-01-01

Anaerobic treatment of cheese whey using a 17.5 litre upflow anaerobic sludge blanket reactor was investigated in the laboratory over a range of influent concentration from 4.5 to 38.1 g COD litre{sup -1} at a constant hydraulic retention time of 5 days. The results indicated that two sludge distribution regions, a sludge bed and a sludge blanket, as well as two distinct reaction phases, acidogenic and methanogenic, were formed. However, as the substrate loading was increased, the acidogenic region extended into the methanogenic region in the upper portion of the reactor until the whole region was acidogenic, leading to the failure of the reactor. (author).

Bacterial population of piggery-waste anaerobic digesters

Energy Technology Data Exchange (ETDEWEB)

Hobson, P N; Shaw, B G

1974-08-01

A survey was made of the anaerobic and facultatively anaerobic bacteria present in piggery waste, digesting piggery waste and domestic anaerobic sludge used to start a piggery waste digester. An influence of the input waste was shown in that streptococci, the predominant facultatively anaerobic bacteria in the piggery waste, were the predominant bacteria in the digesting waste, and they replaced Entrobacter, predominant in the domestic sludge, when a piggery waste digestion had been established from this latter material. Cellulolytic or methanogenic bacteria could not be detected in the piggery waste but populations of these, and other hydrolytic bacteria, became established at different times during the build-up of digestion by gradual addition of piggery waste to water. The bacteria concerned in degradation of the waste constituents were all anaerobes. Production of methane from H/sub 2//CO/sub 2/, formate and butyrate could be detected in mixed cultures from dilutions of digester contents, but the only methanogenic bacterium that could be isolated in pure culture was Methanobacterium formicicum, which uses H/sub 2//CO/sub 2/ or formate only.

Reductive decolourisation of azo dyes by mesophilic and thermophilic methanogenic consortia

NARCIS (Netherlands)

Cervantes, F.J.; Santos, dos A.B.; Madrid, de M.P.; Stams, A.J.M.; Lier, van J.B.

2005-01-01

The contribution of acidogenic bacteria and methanogenic archaea on the reductive decolourisation of azo dyes was assessed in anaerobic granular sludge. Acidogenic bacteria appeared to play an important role in the decolourising processes when glucose was provided as an electron donor; whereas

Genomic analysis of methanogenic archaea reveals a shift towards energy conservation

DEFF Research Database (Denmark)

Gilmore, Sean P.; Henske, John K.; Sexton, Jessica A.

2017-01-01

Background: The metabolism of archaeal methanogens drives methane release into the environment and is critical to understanding global carbon cycling. Methanogenesis operates at a very low reducing potential compared to other forms of respiration and is therefore critical to many anaerobic enviro...

Isolation and characterization of acetate-utilizing anaerobes from a freshwater sediment

NARCIS (Netherlands)

Scholten, J.C.M.; Stams, A.J.M.

2000-01-01

Acetate-degrading anaerobic microorganisms in freshwater sediment were quantified by the most probable number technique. From the highest dilutions a methanogenic, a sulfate-reducing, and a nitrate-reducing microorganism were isolated with acetate as substrate. The methanogen (culture AMPB-Zg) was

Fermentative degradation of polyethylene glycol by a strictly anaerobic, gram-negative, nonsporeforming bacterium, Pelobacter venetianus sp. nov.

Science.gov (United States)

Schink, B; Stieb, M

1983-06-01

The synthetic polyether polyethylene glycol (PEG) with a molecular weight of 20,000 was anaerobically degraded in enrichment cultures inoculated with mud of limnic and marine origins. Three strains (Gra PEG 1, Gra PEG 2, and Ko PEG 2) of rod-shaped, gram-negative, nonsporeforming, strictly anaerobic bacteria were isolated in mineral medium with PEG as the sole source of carbon and energy. All strains degraded dimers, oligomers, and polymers of PEG up to a molecular weight of 20,000 completely by fermentation to nearly equal amounts of acetate and ethanol. The monomer ethylene glycol was not degraded. An ethylene glycol-fermenting anaerobe (strain Gra EG 12) isolated from the same enrichments was identified as Acetobacterium woodii. The PEG-fermenting strains did not excrete extracellular depolymerizing enzymes and were inhibited by ethylene glycol, probably owing to a blocking of the cellular uptake system. PEG, some PEG-containing nonionic detergents, 1,2-propanediol, 1,2-butanediol, glycerol, and acetoin were the only growth substrates utilized of a broad variety of sugars, organic acids, and alcohols. The isolates did not reduce sulfate, sulfur, thiosulfate, or nitrate and were independent of growth factors. In coculture with A. woodii or Methanospirillum hungatei, PEGs and ethanol were completely fermented to acetate (and methane). A marine isolate is described as the type strain of a new species, Pelobacter venetianus sp. nov. Its physiology and ecological significance, as well as the importance and possible mechanism of anaerobic polyether degradation, are discussed.

Low-temperature (10°C) anaerobic digestion of dilute dairy wastewater in an EGSB bioreactor: microbial community structure, population dynamics, and kinetics of methanogenic populations.

Science.gov (United States)

Bialek, Katarzyna; Cysneiros, Denise; O'Flaherty, Vincent

2013-01-01

The feasibility of anaerobic digestion of dairy wastewater at 10°C was investigated in a high height : diameter ratio EGSB reactor. Stable performance was observed at an applied organic loading rate (OLR) of 0.5-2 kg COD m(-3) d(-1) with chemical oxygen demand (COD) removal efficiencies above 85%. When applied OLR increased to values above 2 kg COD m(-3) d(-1), biotreatment efficiency deteriorated, with methanogenesis being the rate-limiting step. The bioreactor recovered quickly (3 days) after reduction of the OLR. qPCR results showed a reduction in the abundance of hydrogenotrophic methanogenic Methanomicrobiales and Methanobacteriales throughout the steady state period followed by a sharp increase in their numbers (111-fold) after the load shock. Specific methanogenic activity and maximum substrate utilising rate (A(max)) of the biomass at the end of trial indicated increased activity and preference towards hydrogenotrophic methanogenesis, which correlated well with the increased abundance of hydrogenotrophic methanogens. Acetoclastic Methanosaeta spp. remained at stable levels throughout the trial. However, increased apparent half-saturation constant (K(m)) at the end of the trial indicated a decrease in the specific substrate affinity for acetate of the sludge, suggesting that Methanosaeta spp., which have high substrate affinity, started to be outcompeted in the reactor.

Isotope fractionation during the anaerobic consumption of acetate by methanogenic and sulfate-reducing microorganisms

Science.gov (United States)

Gövert, D.; Conrad, R.

2009-04-01

During the anaerobic degradation of organic matter in anoxic sediments and soils acetate is the most important substrate for the final step in production of CO2 and/or CH4. Sulfate-reducing bacteria (SRB) and methane-producing archaea both compete for the available acetate. Knowledge about the fractionation of 13C/12C of acetate carbon by these microbial groups is still limited. Therefore, we determined carbon isotope fractionation in different cultures of acetate-utilizing SRB (Desulfobacter postgatei, D. hydrogenophilus, Desulfobacca acetoxidans) and methanogens (Methanosarcina barkeri, M. acetivorans). Including literature values (e.g., Methanosaeta concilii), isotopic enrichment factors (epsilon) ranged between -35 and +2 permil, possibly involving equilibrium isotope effects besides kinetic isotope effects. The values of epsilon were dependent on the acetate-catabolic pathway of the particular microorganism, the methyl or carboxyl position of acetate, and the relative availability or limitation of the substrate acetate. Patterns of isotope fractionation in anoxic lake sediments and rice field soil seem to reflect the characteristics of the microorganisms actively involved in acetate catabolism. Hence, it might be possible using environmental isotopic information to determine the type of microbial metabolism converting acetate to CO2 and/or CH4.

Optimization of up-flow anaerobic sludge blanket reactor for ...

African Journals Online (AJOL)

aghomotsegin

2013-06-05

Jun 5, 2013 ... sludge instead of imported commercial anaerobic granulated sludge. Over the ... biogas, granulated anaerobic sludge, industrial wastewater. ... production of methane by methanogenic bacteria. Compared with other treatment processes, USAB ... effluent collector; 8, gas outlet; 9, gas collector; 10, side-arm ...

Conductive iron oxide minerals accelerate syntrophic cooperation in methanogenic benzoate degradation

Energy Technology Data Exchange (ETDEWEB)

Zhuang, Li; Tang, Jia; Wang, Yueqiang; Hu, Min; Zhou, Shungui, E-mail: sgzhou@soil.gd.cn

2015-08-15

Highlights: • Paddy soil contaminated with benzoate incubated with hematite and magnetite. • Iron oxides addition enhanced methanogenic benzoate degradation by 25–53%. • The facilitated syntrophy might involve direct interspecies electron transfer. • Bacillaceae, Peptococcaceae, and Methanobacterium are potentially involved. - Abstract: 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 CH{sub 4} 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.

Accelerated anaerobic hydrolysis rates under a combination of intermittent aeration and anaerobic conditions

DEFF Research Database (Denmark)

Jensen, T. R.; Lastra Milone, T.; Petersen, G.

2017-01-01

Anaerobic hydrolysis in activated return sludge was investigated in laboratory scale experiments to find if intermittent aeration would accelerate anaerobic hydrolysis rates compared to anaerobic hydrolysis rates under strict anaerobic conditions. The intermittent reactors were set up in a 240 h...... for calculating hydrolysis rates based on soluble COD were compared. Two-way ANOVA with the Bonferroni post-test was performed in order to register any significant difference between reactors with intermittent aeration and strictly anaerobic conditions respectively. The experiment demonstrated a statistically...... significant difference in favor of the reactors with intermittent aeration showing a tendency towards accelerated anaerobic hydrolysis rates due to application of intermittent aeration. The conclusion of the work is thus that intermittent aeration applied in the activated return sludge process (ARP) can...

Application of Pseudomurein Endoisopeptidase to Fluorescence In Situ Hybridization of Methanogens within the Family Methanobacteriaceae▿

Science.gov (United States)

Nakamura, Kohei ; Terada, Takeshi; Sekiguchi, Yuji; Shinzato, Naoya; Meng, Xian-Ying; Enoki, Miho; Kamagata, Yoichi

2006-01-01

In situ detection of methanogens within the family Methanobacteriaceae is sometimes known to be unsuccessful due to the difficulty in permeability of oligonucleotide probes. Pseudomurein endoisopeptidase (Pei), a lytic enzyme that specifically acts on their cell walls, was applied prior to 16S rRNA-targeting fluorescence in situ hybridization (FISH). For this purpose, pure cultured methanogens within this family, Methanobacterium bryantii, Methanobrevibacter ruminantium, Methanosphaera stadtmanae, and Methanothermobacter thermautotrophicus together with a Methanothermobacter thermautotrophicus-containing syntrophic acetate-oxidizing coculture, endosymbiotic Methanobrevibacter methanogens within an anaerobic ciliate, and an upflow anaerobic sludge blanket (UASB) granule were examined. Even without the Pei treatment, Methanobacterium bryantii and Methanothermobacter thermautotrophicus cells are relatively well hybridized with oligonucleotide probes. However, almost none of the cells of Methanobrevibacter ruminantium, Methanosphaera stadtmanae, cocultured Methanothermobacter thermautotrophicus, and the endosymbiotic methanogens and the cells within UASB granule were hybridized. Pei treatment was able to increase the probe hybridization ratio in every specimen, particularly in the specimen that had shown little hybridization. Interestingly, the hybridizing signal intensity of Methanothermobacter thermautotrophicus cells in coculture with an acetate-oxidizing H2-producing syntroph was significantly improved by Pei pretreatment, whereas the probe was well hybridized with the cells of pure culture of the same strain. We found that the difference is attributed to the differences in cell wall thicknesses between the two culture conditions. These results indicate that Pei treatment is effective for FISH analysis of methanogens that show impermeability to the probe. PMID:16950902

A comprehensive study into the molecular methodology and molecular biology of methanogenic Archaea

DEFF Research Database (Denmark)

Lange, M.; Ahring, Birgitte Kiær

2001-01-01

Methanogens belong to the kingdom of Euryarchaeota in the domain of Archaea. The Archaea differ from Bacteria in many aspects important to molecular work. Among these are cell wall composition, their sensitivity to antibiotics, their translation and transcription machinery, and their very strict ...... procedures. Efficient genetic manipulation systems, including shuttle and integration vector systems, have appeared for mesophilic, but not for thermophilic species within the last few years and will have a major impact on future investigations of methanogenic molecular biology....

Early stages in biofilm development in methanogenic fluidized-bed reactors

Energy Technology Data Exchange (ETDEWEB)

Lauwers, A.M.; Heinen, W.; Gorris, L.G.M.; Drift, C. van der (Katholieke Univ. Nijmegen (Netherlands). Dept. of Microbiology and Evolution Biology)

1990-06-01

Biofilm development in methanogenic fluidized-bed reactors with sand as the carrier was studied on a laboratory scale. The microorganisms present in consecutive layers of the biofilm of mature sludge granules were prelimilarily characterized on the basis of their morphology, element composition and adhesion capacity and were compared to bacteria which take part in the initial colonization of sand. The early phase of biofilm development was monitored with reactors receiving waste-waters containing different mixtures of volatile fatty acids and inoculated with fluidized-bed reactor effluent for different lengths of time. The results obtained indicate that facultative anaerobic bacteria abundantly present in the outermost biofilm layers of mature sludge granules are probably the main primary colonizers of the sand. Methanothrix spp. or other methanogens were rarely observed among the primary colonizers. The course of biofilm formation was comparable under the various start-up conditions employed including variations in waste-water composition, inoculation and anaerobicity. However, omission of waste-water and thus of substrate resulted in rapid wash-out of the attached biomass. (orig.).

Establishment of a Methanogenic Benzene-Degrading Culture and its Implication in Bioremediation

Science.gov (United States)

Qiao, W.; Luo, F.; Bawa, N.; Guo, S.; Ye, S.; Edwards, E.

2017-12-01

Benzene is a known human carcinogen and it is a common pollutant in groundwater, mainly resulting from petrochemical industry. Anaerobic degradation of benzene has significant advantages over aerobic processes for in situ bioremediation. In this study, new methanogenic and sulfate-reducing benzene degrading cultures have been enriched. Microbial community composition was characterized with two other previously established benzene-degrading cultures, and their potential use in bioaugmentation is investigated. In this study, a lab microcosm study was conducted anaerobically with contaminated soil and groundwater from a former chemical plant. Benzene degradation was observed in the presence of co-contaminants and electron donor. Through repetitive amendment of benzene, two enrichment cultures have been developed under sulfate and methanogenic conditions. Results from DNA amplicon sequencing and qPCR analysis revealed that an organism similar to previously described benzene-degrading Deltaproteobacterium has been enriched. The microbial community of this culture was compared with other two methanogenic benzene-degrading enrichment cultures that were derived from an oil refinery and a decommissioned gasoline station, and have been maintained for decades. Deltaproteobacterium ORM2-like microbes were dominate in all enrichment cultures, which brought to light benzene-degrading microbes, ORM2 were enriched under different geological conditions distributed around the world. The relative abundance of methanogens was much lower compared to previously established cultures, although substantial amount of methane was produced. The peripheral organisms also vary. To investigate effectiveness of using ORM2-dominant enrichment cultures in bioremediation, microcosm studies were set up using contaminated materials, and a ORM2-dominating methanogenic benzene-degrading culture was used for bioaugmentation. Results revealed that benzene degradation was speeded up under methanogenic or

A simple and sensitive quality control method of the anaerobic atmosphere for identification and antimicrobial susceptibility testing of anaerobic bacteria

DEFF Research Database (Denmark)

Justesen, Tage; Justesen, Ulrik Stenz

2013-01-01

The maintenance of a strict anaerobic atmosphere is essential for the culture of strict anaerobic bacteria. We describe a simple and sensitive quality control method of the anaerobic atmosphere, based on the measurement of the zone diameter around a 5-μg metronidazole disk when testing...... an aerotolerant Clostridium perfringens strain. A zone diameter above 27 mm was indicative of acceptable anaerobic conditions....

Peptidolytic microbial community of methanogenic reactors from two modified UASBs of brewery industries

Directory of Open Access Journals (Sweden)

C. Díaz

2010-10-01

Full Text Available 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 x 10(9 bacteria/g volatile suspended solids VSS (Methanogenic Reactor 1 and 7.2 x 10(6 and 6.4 x 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 d-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.

Rapid establishment of thermophilic anaerobic microbial community during the one-step startup of thermophilic anaerobic digestion from a mesophilic digester.

Science.gov (United States)

Tian, Zhe; Zhang, Yu; Li, Yuyou; Chi, Yongzhi; Yang, Min

2015-02-01

The purpose of this study was to explore how fast the thermophilic anaerobic microbial community could be established during the one-step startup of thermophilic anaerobic digestion from a mesophilic digester. Stable thermophilic anaerobic digestion was achieved within 20 days from a mesophilic digester treating sewage sludge by adopting the one-step startup strategy. The succession of archaeal and bacterial populations over a period of 60 days after the temperature increment was followed by using 454-pyrosequencing and quantitative PCR. After the increase of temperature, thermophilic methanogenic community was established within 11 days, which was characterized by the fast colonization of Methanosarcina thermophila and two hydrogenotrophic methanogens (Methanothermobacter spp. and Methanoculleus spp.). At the same time, the bacterial community was dominated by Fervidobacterium, whose relative abundance rapidly increased from 0 to 28.52 % in 18 days, followed by other potential thermophilic genera, such as Clostridium, Coprothermobacter, Anaerobaculum and EM3. The above result demonstrated that the one-step startup strategy could allow the rapid establishment of the thermophilic anaerobic microbial community. Copyright © 2014 Elsevier Ltd. All rights reserved.

Low-Temperature (10°C Anaerobic Digestion of Dilute Dairy Wastewater in an EGSB Bioreactor: Microbial Community Structure, Population Dynamics, and Kinetics of Methanogenic Populations

Directory of Open Access Journals (Sweden)

Katarzyna Bialek

2013-01-01

Full Text Available The feasibility of anaerobic digestion of dairy wastewater at 10°C was investigated in a high height : diameter ratio EGSB reactor. Stable performance was observed at an applied organic loading rate (OLR of 0.5–2 kg COD m−3 d−1 with chemical oxygen demand (COD removal efficiencies above 85%. When applied OLR increased to values above 2 kg COD m−3 d−1, biotreatment efficiency deteriorated, with methanogenesis being the rate-limiting step. The bioreactor recovered quickly (3 days after reduction of the OLR. qPCR results showed a reduction in the abundance of hydrogenotrophic methanogenic Methanomicrobiales and Methanobacteriales throughout the steady state period followed by a sharp increase in their numbers (111-fold after the load shock. Specific methanogenic activity and maximum substrate utilising rate (Amax of the biomass at the end of trial indicated increased activity and preference towards hydrogenotrophic methanogenesis, which correlated well with the increased abundance of hydrogenotrophic methanogens. Acetoclastic Methanosaeta spp. remained at stable levels throughout the trial. However, increased apparent half-saturation constant (Km at the end of the trial indicated a decrease in the specific substrate affinity for acetate of the sludge, suggesting that Methanosaeta spp., which have high substrate affinity, started to be outcompeted in the reactor.

Physiologically anaerobic microorganisms of the deep subsurface

Energy Technology Data Exchange (ETDEWEB)

Stevens, S.E. Jr.; Chung, K.T.

1992-06-01

A variety of different media were used to isolate facultatively (FAB) and obligately anaerobic bacteria (OAB). These bacteria were isolated from core subsamples obtained from boreholes at the Idaho National Engineering Lab. (INEL) or at the Hanford Lab. (Yakima). Core material was sampled at various depths to 600 feet below the surface. All core samples with culturable bacteria contained at least FAB making thisthe most common physiological type of anaerobic bacteria present in the deep subsurface at these two sites. INEL core samples are characterized by isolates of both FAB and OAB. No isolates of acetogenic, methanogenic, or sulfate reducing bacteria were obtained. Yakima core samples are characterized by a marked predominance of FAB in comparison to OAB. In addition, isolates of acetogenic, methanogenic, and sulfate reducing bacteria were obtained. The Yakima site has the potential for complete anaerobic mineralization of organic compounds whereas this potential appears to be lacking at INEL.

Anaerobic bacteria that dechlorinate perchloroethene.

Science.gov (United States)

Fathepure, B Z; Nengu, J P; Boyd, S A

1987-01-01

In this study, we identified specific cultures of anaerobic bacteria that dechlorinate perchlorethene (PCE). The bacteria that significantly dechlorinated PCE were strain DCB-1, an obligate anaerobe previously shown to dechlorinate chlorobenzoate, and two strains of Methanosarcina. The rate of PCE dechlorination by DCB-1 compared favorably with reported rates of trichloroethene bio-oxidation by methanotrophs. Even higher PCE dechlorination rates were achieved when DCB-1 was grown in a methanogenic consortium. PMID:3426224

Phenotypic properties and microbial diversity of methanogenic granules from a full-scale upflow anaerobic sludge bed reactor treating brewery wastewater.

Science.gov (United States)

Díaz, Emiliano E; Stams, Alfons J M; Amils, Ricardo; Sanz, José L

2006-07-01

Methanogenic granules from an anaerobic bioreactor that treated wastewater of a beer brewery consisted of different morphological types of granules. In this study, the microbial compositions of the different granules were analyzed by molecular microbiological techniques: cloning, denaturing gradient gel electrophoresis and fluorescent in situ hybridization (FISH), and scanning and transmission electron microscopy. We propose here that the different types of granules reflect the different stages in the life cycle of granules. Young granules were small, black, and compact and harbored active cells. Gray granules were the most abundant granules. These granules have a multilayer structure with channels and void areas. The core was composed of dead or starving cells with low activity. The brown granules, which were the largest granules, showed a loose and amorphous structure with big channels that resulted in fractured zones and corresponded to the older granules. Firmicutes (as determined by FISH) and Nitrospira and Deferribacteres (as determined by cloning and sequencing) were the predominant Bacteria. Remarkably, Firmicutes could not be detected in the brown granules. The methanogenic Archaea identified were Methanosaeta concilii (70 to 90% by FISH and cloning), Methanosarcina mazei, and Methanospirillum spp. The phenotypic appearance of the granules reflected the physiological condition of the granules. This may be valuable to easily select appropriate seed sludges to start up other reactors.

Study and optimization of the biodegradation of Polycyclic Aromatic Hydrocarbons (PAHs) and Poly-chloro-biphenyls (PCBs) during the anaerobic and aerobic digestion of long-term contaminated urban sludge; Etude et optimisation de la biodegradation d'hydrocarbures aromatiques polycycliques (HAPs) et de polychlorobiphenyls (PCBs) au cours de la digestion anaerobie et aerobie de boues urbaines contaminees

Energy Technology Data Exchange (ETDEWEB)

Trably, E.

2002-12-15

This study deals with the behavior of PAHs and PCBs during anaerobic and aerobic digestion of long-term contaminated sludge. Initially, an analytical method of 13 PAHs in sludge was developed to PAH-monitoring in laboratory-scaled bioreactors. For this, the method was optimized and validated for its high accuracy and its high reproducibility. In order to estimate precisely the PAH and PCB biological removal performances of each reactor, it was also proposed a method of analysis of the results based on mass balance. Therefore, it was observed for the first time significant PAHs removal under methanogenic conditions. It was also shown that PAH and PCB removals were limited by the mass transfer kinetics and particularly by the reduction of solids. The anaerobic and aerobic processes were then optimized by improving the PAH diffusion with the enhancement of reactor temperature and with the addition of surfactants and solvent, such as methanol. It was highlighted the great fragility of the methanogenic ecosystems and, on the opposite, the strong potential of the aerobic ecosystem for PAHs biodegradation. Indeed, some aerobic processes were successful in decontaminating sludge significantly (at 45 deg. C or in the presence of methanol). Lastly, the PAH biodegradation was characterized partly by the monitoring of {sup 14}C-radiolabelled compounds and by the molecular identification of the methanogenic archaea species. It was suggested that some archaea microorganisms were implied in PAHs biodegradation under strict anaerobic methanogenic conditions. (author)

Early Microbial Evolution: The Age of Anaerobes.

Science.gov (United States)

Martin, William F; Sousa, Filipa L

2015-12-18

In this article, the term "early microbial evolution" refers to the phase of biological history from the emergence of life to the diversification of the first microbial lineages. In the modern era (since we knew about archaea), three debates have emerged on the subject that deserve discussion: (1) thermophilic origins versus mesophilic origins, (2) autotrophic origins versus heterotrophic origins, and (3) how do eukaryotes figure into early evolution. Here, we revisit those debates from the standpoint of newer data. We also consider the perhaps more pressing issue that molecular phylogenies need to recover anaerobic lineages at the base of prokaryotic trees, because O2 is a product of biological evolution; hence, the first microbes had to be anaerobes. If molecular phylogenies do not recover anaerobes basal, something is wrong. Among the anaerobes, hydrogen-dependent autotrophs--acetogens and methanogens--look like good candidates for the ancestral state of physiology in the bacteria and archaea, respectively. New trees tend to indicate that eukaryote cytosolic ribosomes branch within their archaeal homologs, not as sisters to them and, furthermore tend to root archaea within the methanogens. These are major changes in the tree of life, and open up new avenues of thought. Geochemical methane synthesis occurs as a spontaneous, abiotic exergonic reaction at hydrothermal vents. The overall similarity between that reaction and biological methanogenesis fits well with the concept of a methanogenic root for archaea and an autotrophic origin of microbial physiology. Copyright © 2016 Cold Spring Harbor Laboratory Press; all rights reserved.

Bioelectrochemical enhancement of anaerobic methanogenesis for high organic load rate wastewater treatment in a up-flow anaerobic sludge blanket (UASB) reactor.

Science.gov (United States)

Zhao, Zhiqiang; Zhang, Yaobin; Chen, Shuo; Quan, Xie; Yu, Qilin

2014-10-17

A coupling process of anaerobic methanogenesis and electromethanogenesis was proposed to treat high organic load rate (OLR) wastewater. During the start-up stage, acetate removal efficiency of the electric-biological reactor (R1) reached the maximization about 19 percentage points higher than that of the control anaerobic reactor without electrodes (R2), and CH4 production rate of R1 also increased about 24.9% at the same time, while additional electric input was 1/1.17 of the extra obtained energy from methane. Coulombic efficiency and current recorded showed that anodic oxidation contributed a dominant part in degrading acetate when the metabolism of methanogens was low during the start-up stage. Along with prolonging operating time, aceticlastic methanogenesis gradually replaced anodic oxidation to become the main pathway of degrading acetate. When the methanogens were inhibited under the acidic conditions, anodic oxidation began to become the main pathway of acetate decomposition again, which ensured the reactor to maintain a stable performance. FISH analysis confirmed that the electric field imposed could enrich the H2/H(+)-utilizing methanogens around the cathode to help for reducing the acidity. This study demonstrated that an anaerobic digester with a pair of electrodes inserted to form a coupling system could enhance methanogenesis and reduce adverse impacts.

[Long-term storage of obligate anaerobic microorganisms in glycerol].

Science.gov (United States)

Briukhanov, A I; Netrusov, A I

2006-01-01

We evaluated the possibility of storing the cultures of obligate anaerobic microorganisms (clostridia. acetogenic and sulfate-reducing bacteria, and methanogenic archaea) in 25% glycerol at -70 degrees C for a long time (up to 3 years). This method of storage is adequate to preserve cell viability in most obligate anaerobes.

Methanogens in the Solar System

Science.gov (United States)

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

Methanogen community composition and rates of methane consumption in Canadian High Arctic permafrost soils.

Science.gov (United States)

Allan, J; Ronholm, J; Mykytczuk, N C S; Greer, C W; Onstott, T C; Whyte, L G

2014-04-01

Increasing permafrost thaw, driven by climate change, has the potential to result in organic carbon stores being mineralized into carbon dioxide (CO2) and methane (CH4) through microbial activity. This study examines the effect of increasing temperature on community structure and metabolic activity of methanogens from the Canadian High Arctic, in an attempt to predict how warming will affect microbially controlled CH4 soil flux. In situ CO2 and CH4 flux, measured in 2010 and 2011 from ice-wedge polygons, indicate that these soil formations are a net source of CO2 emissions, but a CH4 sink. Permafrost and active layer soil samples were collected at the same sites and incubated under anaerobic conditions at warmer temperatures, with and without substrate amendment. Gas flux was measured regularly and indicated an increase in CH4 flux after extended incubation. Pyrosequencing was used to examine the effects of an extended thaw cycle on methanogen diversity and the results indicate that in situ methanogen diversity, based on the relative abundance of the 16S ribosomal ribonucleic acid (rRNA) gene associated with known methanogens, is higher in the permafrost than in the active layer. Methanogen diversity was also shown to increase in both the active layer and permafrost soil after an extended thaw. This study provides evidence that although High Arctic ice-wedge polygons are currently a sink for CH4, higher arctic temperatures and anaerobic conditions, a possible result of climate change, could result in this soil becoming a source for CH4 gas flux. © 2013 Society for Applied Microbiology and John Wiley & Sons Ltd.

Exocellular electron transfer in anaerobic microbial communities.

Science.gov (United States)

Stams, Alfons J M; de Bok, Frank A M; Plugge, Caroline M; van Eekert, Miriam H A; Dolfing, Jan; Schraa, Gosse

2006-03-01

Exocellular electron transfer plays an important role in anaerobic microbial communities that degrade organic matter. Interspecies hydrogen transfer between microorganisms is the driving force for complete biodegradation in methanogenic environments. Many organic compounds are degraded by obligatory syntrophic consortia of proton-reducing acetogenic bacteria and hydrogen-consuming methanogenic archaea. Anaerobic microorganisms that use insoluble electron acceptors for growth, such as iron- and manganese-oxide as well as inert graphite electrodes in microbial fuel cells, also transfer electrons exocellularly. Soluble compounds, like humic substances, quinones, phenazines and riboflavin, can function as exocellular electron mediators enhancing this type of anaerobic respiration. However, direct electron transfer by cell-cell contact is important as well. This review addresses the mechanisms of exocellular electron transfer in anaerobic microbial communities. There are fundamental differences but also similarities between electron transfer to another microorganism or to an insoluble electron acceptor. The physical separation of the electron donor and electron acceptor metabolism allows energy conservation in compounds as methane and hydrogen or as electricity. Furthermore, this separation is essential in the donation or acceptance of electrons in some environmental technological processes, e.g. soil remediation, wastewater purification and corrosion.

Unexpected competitiveness of Methanosaeta populations at elevated acetate concentrations in methanogenic treatment of animal wastewater.

Science.gov (United States)

Chen, Si; Cheng, Huicai; Liu, Jiang; Hazen, Terry C; Huang, Vicki; He, Qiang

2017-02-01

Acetoclastic methanogenesis is a key metabolic process in anaerobic digestion, a technology with broad applications in biogas production and waste treatment. Acetoclastic methanogenesis is known to be performed by two archaeal genera, Methanosaeta and Methanosarcina. The conventional model posits that Methanosaeta populations are more competitive at low acetate levels (competitiveness of Methanosaeta at elevated acetate was further supported by the enrichment of Methanosaeta with high concentrations of acetate (20 mM). The dominance of Methanosaeta in the methanogen community could be reproduced in anaerobic digesters with the direct addition of acetate to above 20 mM, again supporting the competitiveness of Methanosaeta over Methanosarcina at elevated acetate levels. This study for the first time systematically demonstrated that the dominance of Methanosaeta populations in anaerobic digestion could be linked to the competitiveness of Methanosaeta at elevated acetate concentrations. Given the importance of acetoclastic methanogenesis in biological methane production, findings from this study could have major implications for developing strategies for more effective control of methanogenic treatment processes.

The effect of operational conditions on the sludge specific methanogenic activity and sludge biodegradability

NARCIS (Netherlands)

Leitao, R.; Santaellla, S.T.; Haandel, van A.C.; Zeeman, G.; Lettinga, G.

2009-01-01

The effects of hydraulic retention time (HRT) and influent COD concentration (CODInf) on Specific Methanogenic Activity (SMA) and the biodegradability of an anaerobic sludge need to be elucidated because of the discordant results available in literature. This information is important for the

Computational Modeling of Fluctuations in Energy and Metabolic Pathways of Methanogenic Archaea

Energy Technology Data Exchange (ETDEWEB)

Luthey-Schulten, Zaida [Univ. of Illinois, Urbana-Champaign, IL (United States). Dept. of Chemistry; Carl R. Woese Inst. for Genomic Biology

2017-01-04

The methanogenic archaea, anaerobic microbes that convert CO2 and H2 and/or other small organic fermentation products into methane, play an unusually large role in the global carbon cycle. As they perform the final step in the anaerobic breakdown of biomass, methanogens are a biogenic source of an estimated one billion tons methane each year. Depending on the location, produced methane can be considered as either a greenhouse gas (agricultural byproduct), sequestered carbon storage (methane hydrate deposits), or a potential energy source (organic wastewater treatment). These microbes therefore represent an important target for biotechnology applications. Computational models of methanogens with predictive power are useful aids in the adaptation of methanogenic systems, but need to connect processes of wide-ranging time and length scales. In this project, we developed several computational methodologies for modeling the dynamic behavior of entire cells that connects stochastic reaction-diffusion dynamics of individual biochemical pathways with genome-scale modeling of metabolic networks. While each of these techniques were in the realm of well-defined computational methods, here we integrated them to develop several entirely new approaches to systems biology. The first scientific aim of the project was to model how noise in a biochemical pathway propagates into cellular phenotypes. Genetic circuits have been optimized by evolution to regulate molecular processes despite stochastic noise, but the effect of such noise on a cellular biochemical networks is currently unknown. An integrated stochastic/systems model of Escherichia coli species was created to analyze how noise in protein expression gives—and therefore noise in metabolic fluxes—gives rise to multiple cellular phenotype in isogenic population. After the initial work developing and validating methods that allow characterization of the heterogeneity in the model organism E. coli, the project shifted toward

Treatment of industrial wastewaters by anaerobic membrane bioreactors : Implications of substrate characteristics

NARCIS (Netherlands)

Dereli, R.K.

2015-01-01

The success of anaerobic digestion relies on the presence of highly active methanogenic biomass, requiring effective retention of slow growing anaerobic microorganisms inside bioreactor by decoupling the hydraulic retention time (HRT) from solids residence time (SRT) or the employment of long SRTs

Carbon nanotubes accelerate methane production in pure cultures of methanogens and in a syntrophic coculture

NARCIS (Netherlands)

Salvador, Andreia F.; Martins, Gilberto; Melle-Franco, Manuel; Serpa, Ricardo; Stams, Alfons J.M.; Cavaleiro, Ana J.; Pereira, M.A.; Alves, M.M.

2017-01-01

Carbon materials have been reported to facilitate direct interspecies electron transfer (DIET) between bacteria and methanogens improving methane production in anaerobic processes. In this work, the effect of increasing concentrations of carbon nanotubes (CNT) on the activity of pure cultures of

Exocellular electron transfer in anaerobic microbial communities

NARCIS (Netherlands)

Stams, A.J.M.; Bok, de F.A.M.; Plugge, C.M.; Eekert, van M.H.A.; Dolfing, J.; Schraa, G.

2006-01-01

Exocellular electron transfer plays an important role in anaerobic microbial communities that degrade organic matter. Interspecies hydrogen transfer between microorganisms is the driving force for complete biodegradation in methanogenic environments. Many organic compounds are degraded by obligatory

Evaluation of support matrices for immobilization of anaerobic consortia for efficient carbon cycling in waste regeneration.

Science.gov (United States)

Chauhan, Ashvini; Ogram, Andrew

2005-02-18

Efficient metabolism of fatty acids during anaerobic waste digestion requires development of consortia that include "fatty acid consuming H(2) producing bacteria" and methanogenic bacteria. The objective of this research was to optimize methanogenesis from fatty acids by evaluating a variety of support matrices for use in maintaining efficient syntrophic-methanogenic consortia. Tested matrices included clays (montmorillonite and bentonite), glass beads (106 and 425-600mum), microcarriers (cytopore, cytodex, cytoline, and cultispher; conventionally employed for cultivation of mammalian cell lines), BioSep beads (powdered activated carbon), and membranes (hydrophilic; nylon, polysulfone, and hydrophobic; teflon, polypropylene). Data obtained from headspace methane (CH(4)) analyses as an indicator of anaerobic carbon cycling efficiency indicated that material surface properties were important in maintenance and functioning of the anaerobic consortia. Cytoline yielded significantly higher CH(4) than other matrices as early as in the first week of incubation. 16S rRNA gene sequence analysis from crushed cytoline matrix showed the presence of Syntrophomonas spp. (butyrate oxidizing syntrophs) and Syntrophobacter spp. (propionate oxidizing syntrophs), with Methanosaeta spp. (acetate utilizing methanogen), and Methanospirillum spp. (hydrogen utilizing methanogen) cells. It is likely that the more hydrophobic surfaces provided a suitable surface for adherence of cells of syntrophic-methanogenic consortia. Cytoline also appeared to protect entrapped consortia from air, resulting in rapid methanogenesis after aerial exposure. Our study suggests that support matrices can be used in anaerobic digestors, pre-seeded with immobilized or entrapped consortia on support matrices, and may be of value as inoculant-adsorbents to rapidly initiate or recover proper system functioning following perturbation.

Anaerobic treatment of complex wastewater and waste activated sludge - Appl. of an upflow anaerobic solid removal (UASR).

NARCIS (Netherlands)

Zeeman, G.; Sanders, W.T.M.; Wang, K.Y.; Lettinga, G.

1997-01-01

The application of one phase anaerobic wastewater systems for the treatment of complex wastewaters containing high amounts of suspended solids or lipids is usually limited by accumulation of these compounds in the sludge bed. This accumulation reduces the solid retention time and methanogenic

Anaerobic digestion of food waste stabilized by lime mud from papermaking process.

Science.gov (United States)

Zhang, Jishi; Wang, Qinqing; Zheng, Pengwei; Wang, Yusong

2014-10-01

The effects of lime mud from papermaking process (LMP) addition as buffer agent and inorganic nutrient on the anaerobic digestion stability of food waste (FW) were investigated under mesophilic conditions with the aim of avoiding volatile fatty acids accumulation, and inorganic elements deficiency. When LMP concentration ranged from 6.0 to 10g/L, the FW anaerobic digestion could maintain efficient and stable state. These advantages are attributed to the existence of Ca, Na, Mg, K, Fe, and alkaline substances that favor the methanogenic process. The highest CH4 yield of 272.8mL/g-VS was obtained at LMP and VS concentrations of 10.0 and 19.8g/L, respectively, with the corresponding lag-phase time of 3.84d and final pH of 8.4. The methanogens from residue digestates mainly consisted of Methanobrevibacter, coccus-type and sarcina-type methanogens with LMP addition compared to Methanobacteria in control. However, higher concentration of LMP inhibited methanogenic activities and methane production. Copyright © 2014 Elsevier Ltd. All rights reserved.

Syntrophic acetate oxidation in two-phase (acid-methane) anaerobic digesters.

Science.gov (United States)

Shimada, T; Morgenroth, E; Tandukar, M; Pavlostathis, S G; Smith, A; Raskin, L; Kilian, R E

2011-01-01

The microbial processes involved in two-phase anaerobic digestion were investigated by operating a laboratory-scale acid-phase (AP) reactor and analyzing two full-scale, two-phase anaerobic digesters operated under mesophilic (35 °C) conditions. The digesters received a blend of primary sludge and waste activated sludge (WAS). Methane levels of 20% in the laboratory-scale reactor indicated the presence of methanogenic activity in the AP. A phylogenetic analysis of an archaeal 16S rRNA gene clone library of one of the full-scale AP digesters showed that 82% and 5% of the clones were affiliated with the orders Methanobacteriales and Methanosarcinales, respectively. These results indicate that substantial levels of aceticlastic methanogens (order Methanosarcinales) were not maintained at the low solids retention times and acidic conditions (pH 5.2-5.5) of the AP, and that methanogenesis was carried out by hydrogen-utilizing methanogens of the order Methanobacteriales. Approximately 43, 31, and 9% of the archaeal clones from the methanogenic phase (MP) digester were affiliated with the orders Methanosarcinales, Methanomicrobiales, and Methanobacteriales, respectively. A phylogenetic analysis of a bacterial 16S rRNA gene clone library suggested the presence of acetate-oxidizing bacteria (close relatives of Thermacetogenium phaeum, 'Syntrophaceticus schinkii,' and Clostridium ultunense). The high abundance of hydrogen consuming methanogens and the presence of known acetate-oxidizing bacteria suggest that acetate utilization by acetate oxidizing bacteria in syntrophic interaction with hydrogen-utilizing methanogens was an important pathway in the second-stage of the two-phase digestion, which was operated at high ammonium-N concentrations (1.0 and 1.4 g/L). A modified version of the IWA Anaerobic Digestion Model No. 1 (ADM1) with extensions for syntrophic acetate oxidation and weak-acid inhibition adequately described the dynamic profiles of volatile acid production

Methanogenic degradation of acetone by an enrichment culture

OpenAIRE

Platen, Harald; Schink, Bernhard

1987-01-01

An anaerobic enrichment culture degraded 1 mol of acetone to 2 tool of methane and 1 tool of carbon dioxide. Two microorganisms were involved in this process, a filament-forming rod similar to Methanothrix sp. and an unknown rod with round to slightly pointed ends. Both organisms formed aggregates up to 300 gm in diameter. No fluorescing bacteria were observed indicating that hydrogen or formate-utilizing methanogens are not involved in this process. Acetate was utilized in this culture by th...

The anaerobic treatment of sulfate containing wastewater

NARCIS (Netherlands)

Visser, A.

1995-01-01

In the anaerobic treatment of sulfate containing wastewater sulfate reducing bacteria (SRB) will compete with methanogenic- (MB) and acetogenic bacteria (AB) for the available substrates such as hydrogen, acetate, propionate and butyrate. The outcome of this competition will

The fate of methanol in anaerobic bioreactors

NARCIS (Netherlands)

Florencio, L.

1994-01-01

Methanol is an important component of certain industrial wastewaters. In anaerobic environments, methanol can be utilized by methanogens and acetogens. In wastewater treatment plants, the conversion of methanol into methane is preferred because this conversion is responsible for chemical

The effect of operational conditions on the sludge specific methanogenic activity and sludge biodegradability

International Nuclear Information System (INIS)

Leitao, R. C.; Santaella, S. T.; Haandel, A. C. van; Zeeman, G.; Lettinga, G.

2009-01-01

The Specific Methanogenic Activity (SMA) and sludge biodegradability of an anaerobic sludge depends on various operational and environmental conditions imposed to the anaerobic reactor. However, the effects of hydraulic retention time (HRT), influent COD concentration (COD i nf) and sludge retention time (SRT) on those two parameters need to be elucidated. This knowledge about SMA can provide insights about the capacity of the UASB reactors to withstand organic and hydraulic shock loads, whereas the biodegradability gives information necessary for final disposal of the sludge. (Author)

The effect of operational conditions on the sludge specific methanogenic activity and sludge biodegradability

Energy Technology Data Exchange (ETDEWEB)

Leitao, R. C.; Santaella, S. T.; Haandel, A. C. van; Zeeman, G.; Lettinga, G.

2009-07-01

The Specific Methanogenic Activity (SMA) and sludge biodegradability of an anaerobic sludge depends on various operational and environmental conditions imposed to the anaerobic reactor. However, the effects of hydraulic retention time (HRT), influent COD concentration (COD{sub i}nf) and sludge retention time (SRT) on those two parameters need to be elucidated. This knowledge about SMA can provide insights about the capacity of the UASB reactors to withstand organic and hydraulic shock loads, whereas the biodegradability gives information necessary for final disposal of the sludge. (Author)

Ecophysiology of Uncultured Filamentous Anaerobes Belonging to the Phylum KSB3 That Cause Bulking in Methanogenic Granular Sludge▿ †

Science.gov (United States)

Yamada, Takeshi; Kikuchi, Kae; Yamauchi, Toshihiro; Shiraishi, Koji; Ito, Tsukasa; Okabe, Satoshi; Hiraishi, Akira; Ohashi, Akiyoshi; Harada, Hideki; Kamagata, Yoichi; Nakamura, Kazunori; Sekiguchi, Yuji

2011-01-01

A filamentous bulking of a methanogenic granular sludge caused by uncultured filamentous bacteria of the candidate phylum KSB3 in an upflow anaerobic sludge blanket (UASB) system has been reported. To characterize the physiological traits of the filaments, a polyphasic approach consisting of rRNA-based activity monitoring of the KSB3 filaments using the RNase H method and substrate uptake profiling using microautoradiography combined with fluorescence in situ hybridization (MAR-FISH) was conducted. On the basis of rRNA-based activity, the monitoring of a full-scale UASB reactor operated continuously revealed that KSB3 cells became active and predominant (up to 54% of the total 16S rRNA) in the sludge when the carbohydrate loading to the system increased. Batch experiments with a short incubation of the sludge with maltose, glucose, fructose, and maltotriose at relatively low concentrations (approximately 0.1 mM) in the presence of yeast extract also showed an increase in KSB3 rRNA levels under anaerobic conditions. MAR-FISH confirmed that the KSB3 cells took up radioisotopic carbons from [14C]maltose and [14C]glucose under the same incubation conditions in the batch experiments. These results suggest that one of the important ecophysiological characteristics of KSB3 cells in the sludge is carbohydrate degradation in wastewater and that high carbohydrate loadings may trigger an outbreak of KSB3 bacteria, causing sludge bulking in UASB systems. PMID:21257808

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

Science.gov (United States)

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.

Phenotypic properties and microbial diversity of methanogenic granules from a full-scale UASB reactor treating brewery wastewater

NARCIS (Netherlands)

Diaz, E.E.; Stams, A.J.M.; Amils, R.; Sanz, J.L.

2006-01-01

Methanogenic granules from an anaerobic bioreactor that treated wastewater of a beer brewery consisted of different morphological types of granules. In this study, the microbial compositions of the different granules were analyzed by molecular microbiological techniques: cloning, denaturing gradient

Identification of Anaerobic Aniline-Degrading Bacteria at a Contaminated Industrial Site.

Science.gov (United States)

Sun, Weimin; Li, Yun; McGuinness, Lora R; Luo, Shuai; Huang, Weilin; Kerkhof, Lee J; Mack, E Erin; Häggblom, Max M; Fennell, Donna E

2015-09-15

Anaerobic aniline biodegradation was investigated under different electron-accepting conditions using contaminated canal and groundwater aquifer sediments from an industrial site. Aniline loss was observed in nitrate- and sulfate-amended microcosms and in microcosms established to promote methanogenic conditions. Lag times of 37 days (sulfate amended) to more than 100 days (methanogenic) were observed prior to activity. Time-series DNA-stable isotope probing (SIP) was used to identify bacteria that incorporated (13)C-labeled aniline in the microcosms established to promote methanogenic conditions. In microcosms from heavily contaminated aquifer sediments, a phylotype with 92.7% sequence similarity to Ignavibacterium album was identified as a dominant aniline degrader as indicated by incorporation of (13)C-aniline into its DNA. In microcosms from contaminated canal sediments, a bacterial phylotype within the family Anaerolineaceae, but without a match to any known genus, demonstrated the assimilation of (13)C-aniline. Acidovorax spp. were also identified as putative aniline degraders in both of these two treatments, indicating that these species were present and active in both the canal and aquifer sediments. There were multiple bacterial phylotypes associated with anaerobic degradation of aniline at this complex industrial site, which suggests that anaerobic transformation of aniline is an important process at the site. Furthermore, the aniline degrading phylotypes identified in the current study are not related to any known aniline-degrading bacteria. The identification of novel putative aniline degraders expands current knowledge regarding the potential fate of aniline under anaerobic conditions.

Bacterial study of the anaerobic bioreactor for distillery effluent

International Nuclear Information System (INIS)

Shah, F. A.; Pathan, M. I.

2006-01-01

This study relates with anaerobic bioreactors of Habib Sugar Mills, Nawabshah. Bacterial growth was studied through microscope along with its effect on the production of methane gas (Biogas) at all HRTs (Hydraulic Retention Times) between 15 and 28 days. The bacterium has the efficiency to convert 12% glucose within 24 hours to final product and cell mass. The acetogenic organisms also show their maximum growth on glucose in BGP-1 and BPG-2 at both the corks, where as Methanogenic organisms have shown their zero shown their zero growth on glucose. The efforts have been taken to determine the methanogenic, acetogenic and syntrophomonas sp. data of anaerobic bioreactors of BGP (Biogas Plant) I and II, when these samples were cultured on acetate, methanol, formate, butyrate, propionate and glucose. (author)

High-rate continuous hydrogen production by Thermoanaerobacterium thermosaccharolyticum PSU-2 immobilized on heat-pretreated methanogenic granules

Energy Technology Data Exchange (ETDEWEB)

O-Thong, Sompong [Department of Environmental Engineering, Technical University of Denmark, Bygningstorvet 115, DK-2800, Kgs Lyngby (Denmark); Department of Biology, Faculty of Science, Thaksin University, Patthalung 93110 (Thailand); Prasertsan, Poonsuk [Department of Industrial Biotechnology, Faculty of Agro-Industry, Prince of Songkla University, Hat-Yai, Songkhla 90120 (Thailand); Karakashev, Dimitar; Angelidaki, Irini [Department of Environmental Engineering, Technical University of Denmark, Bygningstorvet 115, DK-2800, Kgs Lyngby (Denmark)

2008-11-15

Biohydrogen production from Thermoanaerobacterium thermosaccharolyticum strain PSU-2 was examined in upflow anaerobic sludge blanket (UASB) reactor and carrier-free upflow anaerobic reactor (UA), both fed with sucrose and operating at 60 C. Heat-pretreated methanogenic granules were used as carrier to immobilize T. thermosaccharolyticum strain PSU-2 in UASB reactor operated at a hydraulic retention time (HRT) ranging from 0.75 to 24 h and corresponding sucrose loading rate from 58.5 to 2.4 mmol sucrose l{sup -1} h{sup -1}. In comparison with hydrogen production rate of 12.1 mmol H{sub 2} l{sup -1} h{sup -1} obtained by carrier-free reactor upflow anaerobic (UA) system, a greatly improved hydrogen production rate up to 152 mmol H{sub 2} l{sup -1} h{sup -1} was demonstrated by the granular cells in UASB system. The biofilm of T. thermosaccharolyticum strain PSU-2 developed on treated methanogenic granules in UASB reactor substantially enhanced biomass retention (3 times), and production of hydrogen (12 times) compared to carrier-free reactor. It appears to be the most preferred process for highly efficient dark fermentative hydrogen production from sugar containing wastewater under thermophilic conditions. (author)

Trace methane oxidation and the methane dependency of sulfate reduction in anaerobic granular sludge

KAUST Repository

Meulepas, Roel J.W.

2010-05-01

This study investigates the oxidation of labeled methane (CH4) and the CH4 dependence of sulfate reduction in three types of anaerobic granular sludge. In all samples, 13C-labeled CH4 was anaerobically oxidized to 13C-labeled CO2, while net endogenous CH4 production was observed. Labeled-CH4 oxidation rates followed CH4 production rates, and the presence of sulfate hampered both labeled-CH4 oxidation and methanogenesis. Labeled-CH4 oxidation was therefore linked to methanogenesis. This process is referred to as trace CH4 oxidation and has been demonstrated in methanogenic pure cultures. This study shows that the ratio between labeled-CH4 oxidation and methanogenesis is positively affected by the CH4 partial pressure and that this ratio is in methanogenic granular sludge more than 40 times higher than that in pure cultures of methanogens. The CH4 partial pressure also positively affected sulfate reduction and negatively affected methanogenesis: a repression of methanogenesis at elevated CH4 partial pressures confers an advantage to sulfate reducers that compete with methanogens for common substrates, formed from endogenous material. The oxidation of labeled CH 4 and the CH4 dependence of sulfate reduction are thus not necessarily evidence of anaerobic oxidation of CH4 coupled to sulfate reduction. © 2010 Federation of European Microbiological Societies.

Biomethane potential of industrial paper wastes and investigation of the methanogenic communities involved.

Science.gov (United States)

Walter, Andreas; Silberberger, Sandra; Juárez, Marina Fernández-Delgado; Insam, Heribert; Franke-Whittle, Ingrid H

2016-01-01

Cellulose-containing waste products from the agricultural or industrial sector are potentially one of the largest sources of renewable energy on earth. In this study, the biomethane potential (BMP) of two types of industrial paper wastes, wood and pulp residues (WR and PR, respectively), were evaluated under both mesophilic and thermophilic conditions, and various pretreatment methods were applied in the attempt to increase the methane potential during anaerobic digestion. The methanogenic community composition was investigated with denaturing gradient gel electrophoresis (DGGE) and the ANAEROCHIP microarray, and dominant methanogens were quantitated using quantitative PCR. All pretreatments investigated in this study with the exception of the alkaline pretreatment of PR were found to increase the BMP of two paper industry wastes. However, the low recalcitrance level of the PR resulted in the pretreatments being less effective in increasing BMP when compared with those for WR. These results were supported by the physico-chemical data. A combined application of ultrasound and enzymatic pretreatment was found to be the best strategy for increasing methane yields. The retention time of substrates in the reactors strongly influenced the BMP of wastes subjected to the different pretreatments. In sludges from both paper wastes subjected to the various pretreatments, mixotrophic Methanosarcina species were found to dominate the community, accompanied by a consortium of hydrogenotrophic genera. Pretreating industrial paper wastes could be a potentially viable option for increasing the overall degradation efficiency and decreasing reactor retention time for the digestion of complex organic matter such as lignocellulose or hemicellulose. This would help reduce the environmental burden generated from paper production. Although there were minor differences in the methanogenic communities depending on the temperature of anaerobic digestion, there was little effect of substrate

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

Science.gov (United States)

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. © 2016 Society for Applied Microbiology and John Wiley & Sons Ltd.

Start-up strategies for thermophilic anaerobic digestion of pig manure

International Nuclear Information System (INIS)

Moset, V.; Bertolini, E.; Cerisuelo, A.; Cambra, M.; Olmos, A.; Cambra-López, M.

2014-01-01

Sludge physicochemical composition, methane (CH 4 ) yield, and methanogenic community structure and dynamics using quantitative real-time polymerase chain reaction were determined after start-up of anaerobic digestion of pig manure. Eight thermophilic continuous stirred anaerobic digesters were used during 126 days. Four management strategies were investigated: a feedless and a non-feedless period followed by a gradual or an abrupt addition of pig manure (two digesters per strategy). During the first 43 days, VFA (volatile fatty acids) accumulations and low CH 4 yield were observed in all digesters. After this period, digesters recovered their initial status being propionic acid the last parameter to be re-established. Non-feedless digesters with an abrupt addition of pig manure showed the best performances (lower VFA accumulation and higher CH 4 yield). Differences in microbial orders and dynamics, however, were less evident among treatments. Hydrogenotrophic methanogenesis, Methanomicrobiales first and Methanobacteriales second, was the dominant metabolic pathway in all digesters. Further research is needed to clarify the role and activity of hydrogenotrophic methanogens during the recovery start-up period and to identify the best molecular tools and methodologies to monitor microbial populations and dynamics reliably and accurately in anaerobic digesters. - Highlights: • Four start-up strategies for thermophilic anaerobic digestion of pig manure were tested. • Physicochemical composition, methane yield and methanogenic community were determined. • During the first 43 days, a decline in reactor's performance occurred. • The best start-up strategy was non-feedless with an abrupt addition of pig slurry. • Hydrogenotrophic methanogenesis was the dominant metabolic pathway

Effect of different ammonia sources on aceticlastic and hydrogenotrophic methanogens

DEFF Research Database (Denmark)

Tian, Hailin; Fotidis, Ioannis; Kissas, Konstantinos

2018-01-01

Ammonium chloride (NH4Cl) was usually used as a model ammonia source to simulate ammonia inhibition during anaerobic digestion (AD) of nitrogen-rich feedstocks. However, ammonia in AD originates mainly from degradation of proteins, urea and nucleic acids, which is distinct from NH4Cl. Thus......, in this study, the inhibitory effect of a “natural” ammonia source (urea) and NH4Cl, on four pure methanogenic strains (aceticlastic: Methanosarcina thermophila, Methanosarcina barkeri; hydrogenotrophic: Methanoculleus bourgensis, Methanoculleus thermophilus), was assessed under mesophilic (37 °C......) and thermophilic (55 °C) conditions. The results showed that urea hydrolysis increased pH significantly to unsuitable levels for methanogenic growth, while NH4Cl had a negligible effect on pH. After adjusting initial pH to 7 and 8, urea was significantly stronger inhibitor with longer lag phases to methanogenesis...

[3H] Thymidine incorporation to estimate growth rates of anaerobic bacterial strains

International Nuclear Information System (INIS)

Winding, A.

1992-01-01

The incorporation of [ 3 H] thymidine by axenic cultures of anaerobic bacteria was investigated as a means to measure growth. The three fermentative strains and one of the methanogenic strains tested incorporated [ 3 H] thymidine during growth. It is concluded that the [ 3 H] thymidine incorporation method underestimates bacterial growth in anaerobic environments

A six-well plate method: less laborious and effective method for cultivation of obligate anaerobic microorganisms.

Science.gov (United States)

Nakamura, Kohei; Tamaki, Hideyuki; Kang, Myung Suk; Mochimaru, Hanako; Lee, Sung-Taik; Nakamura, Kazunori; Kamagata, Yoichi

2011-01-01

We developed a simple, less laborious method to cultivate and isolate obligate anaerobic microorganisms using a six-well plate together with the AnaeroPack System, designated as the six-well plate method. The cultivation efficiency of this method, based on colony-forming units, colony formation time, and colony size, was evaluated with four authentic obligate anaerobes (two methanogenic archaea and two sulfate-reducing bacteria). The method was found to be comparable to or even better than the roll tube method, a technique that is commonly used at present for the cultivation of obligate anaerobes. Further experiments using 21 representative obligate anaerobes demonstrated that all examined anaerobes (11 methanogens, 5 sulfate- or thiosulfate-reducing bacteria, and 5 syntrophs) could form visible colonies on the six-well plate and that these colonies could be successfully subcultured in fresh liquid media. Using this method, an unidentified sulfate-reducing bacterium was successfully isolated from an environmental sample.

Electrochemical monitoring of ammonia during anaerobic digestion

DEFF Research Database (Denmark)

Zhao, Nannan; Angelidaki, Irini; Zhang, Yifeng

Ammonia is known as key inhibitor to methanogens in anaerobic digestion (AD) process. It’s of importance to develop efficient tool for ammonia monitoring. In this study, an electrolysis cell (EC) coupled with a complete nitrification reactor was developed as sensor for real time and online monito...

Biotransformation of tetrachloroethylene to trichloroethylene, dichloroethylene, vinyl chloride, and carbon dioxide under methanogenic conditions

International Nuclear Information System (INIS)

Vogel, T.M.; McCarty, P.L.

1985-01-01

Tetrachloroethylene (PCE) and trichloroethylene (TCE), common industrial solvents, are among the most frequent contaminants found in groundwater supplies. Due to the potential toxicity and carcinogenicity of chlorinated ethylenes, knowledge about their transformation potential is important in evaluating their environmental fate. The results of this study confirm that PCE can be transformed by reductive dehalogenation to TCE, dichloroethylene, and vinyl chloride (VC) under anaerobic conditions. In addition, [ 14 C]PCE was at least partially mineralized to CO 2 . Mineralization of 24% of the PCE occurred in a continuous-flow fixed-film methanogenic column with a liquid detention time of 4 days. TCE was the major intermediate formed, but traces of dichloroethylene isomers and VC were also found. In other column studies under a different set of methanogenic conditions, nearly quantitative conversion of PCE to VC was found. These studies clearly demonstrate that TCE and VC are major intermediates in PCE biotransformation under anaerobic conditions and suggest that the potential exists for the complete mineralization of PCE to CO 2 in soil and aquifer systems and in biological treatment processes

Improving the cyanide toxicity tolerance of anaerobic reactor: Microbial interactions and toxin reduction

International Nuclear Information System (INIS)

Gupta, Pragya; Ahammad, S.Z.; Sreekrishnan, T.R.

2016-01-01

Highlights: • Anaerobic batch study of 110 days. • Acclimatization for cyanide biodegradation. • Understanding inhibitory effects of cyanide on methane generation and VFA production. • Identification of microorganisms tolerant to cyanide. • Community analysis using DGGE and qPCR analyses. - Abstract: Anaerobic biological treatment of high organics containing wastewater is amongst the preferred treatment options but poor tolerance to toxins makes its use prohibitive. In this study, efforts have been made to understand the key parameters for developing anaerobic reactor, resilient to cyanide toxicity. A laboratory scale anaerobic batch reactor was set up to treat cyanide containing wastewater. The reactor was inoculated with anaerobic sludge obtained from a wastewater treatment plant and fresh cow dung in the ratio of 3:1. The focus was on acclimatization and development of cyanide-degrading biomass and to understand the toxic effects of cyanide on the dynamic equilibrium between various microbial groups. The sludge exposed to cyanide was found to have higher bacterial diversity than the control. It was observed that certain hydrogenotrophic methanogens and bacterial groups were able to grow and produce methane in the presence of cyanide. Also, it was found that hydrogen utilizing methanogens were more cyanide tolerant than acetate utilizing methanogens. So, effluents from various industries like electroplating, coke oven plant, petroleum refining, explosive manufacturing, and pesticides industries which are having high concentrations of cyanide can be treated by favoring the growth of the tolerant microbes in the reactors. It will provide much better treatment efficiency by overcoming the inhibitory effects of cyanide to certain extent.

Improving the cyanide toxicity tolerance of anaerobic reactor: Microbial interactions and toxin reduction

Energy Technology Data Exchange (ETDEWEB)

Gupta, Pragya; Ahammad, S.Z.; Sreekrishnan, T.R., E-mail: sree@iitd.ac.in

2016-09-05

Highlights: • Anaerobic batch study of 110 days. • Acclimatization for cyanide biodegradation. • Understanding inhibitory effects of cyanide on methane generation and VFA production. • Identification of microorganisms tolerant to cyanide. • Community analysis using DGGE and qPCR analyses. - Abstract: Anaerobic biological treatment of high organics containing wastewater is amongst the preferred treatment options but poor tolerance to toxins makes its use prohibitive. In this study, efforts have been made to understand the key parameters for developing anaerobic reactor, resilient to cyanide toxicity. A laboratory scale anaerobic batch reactor was set up to treat cyanide containing wastewater. The reactor was inoculated with anaerobic sludge obtained from a wastewater treatment plant and fresh cow dung in the ratio of 3:1. The focus was on acclimatization and development of cyanide-degrading biomass and to understand the toxic effects of cyanide on the dynamic equilibrium between various microbial groups. The sludge exposed to cyanide was found to have higher bacterial diversity than the control. It was observed that certain hydrogenotrophic methanogens and bacterial groups were able to grow and produce methane in the presence of cyanide. Also, it was found that hydrogen utilizing methanogens were more cyanide tolerant than acetate utilizing methanogens. So, effluents from various industries like electroplating, coke oven plant, petroleum refining, explosive manufacturing, and pesticides industries which are having high concentrations of cyanide can be treated by favoring the growth of the tolerant microbes in the reactors. It will provide much better treatment efficiency by overcoming the inhibitory effects of cyanide to certain extent.

Assessment of microbial communities associated with fermentative-methanogenic biodegradation of aromatic hydrocarbons in groundwater contaminated with a biodiesel blend (B20).

Science.gov (United States)

Ramos, Débora Toledo; da Silva, Márcio Luís Busi; Nossa, Carlos Wolfgang; Alvarez, Pedro J J; Corseuil, Henry Xavier

2014-09-01

A controlled field experiment was conducted to assess the potential for fermentative-methanogenic biostimulation (by ammonium-acetate injection) to enhance biodegradation of benzene, toluene, ethylbenzene and xylenes (BTEX) as well as polycyclic aromatic hydrocarbons (PAHs) in groundwater contaminated with biodiesel B20 (20:80 v/v soybean biodiesel and diesel). Changes in microbial community structure were assessed by pyrosequencing 16S rRNA analyses. BTEX and PAH removal began 0.7 year following the release, concomitantly with the increase in the relative abundance of Desulfitobacterium and Geobacter spp. (from 5 to 52.7 % and 15.8 to 37.3 % of total Bacteria 16S rRNA, respectively), which are known to anaerobically degrade hydrocarbons. The accumulation of anaerobic metabolites acetate and hydrogen that could hinder the thermodynamic feasibility of BTEX and PAH biotransformations under fermentative/methanogenic conditions was apparently alleviated by the growing predominance of Methanosarcina. This suggests the importance of microbial population shifts that enrich microorganisms capable of interacting syntrophically to enhance the feasibility of fermentative-methanogenic bioremediation of biodiesel blend releases.

Variation of carbon isotope fractionation in hydrogenotrophic methanogenic microbial cultures and environmental samples at different energy status

NARCIS (Netherlands)

Penning, H.; Plugge, C.M.; Galand, P.E.; Conrad, R.

2005-01-01

Methane is a major product of anaerobic degradation of organic matter and an important greenhouse gas. Its stable carbon isotope composition can be used to reveal active methanogenic pathways, if associated isotope fractionation factors are known. To clarify the causes that lead to the wide

Fermentation of glycolate by a pure culture of a strictly anaerobic gram-positive bacterium belonging to the family Lachnospiraceae.

Science.gov (United States)

Janssen, Peter H; Hugenholtz, Philip

2003-05-01

The component bacteria of a three-membered mixed culture able to ferment glycolate to acetate, propionate and CO(2) were isolated in pure culture. All three strains were strict anaerobes that, on the basis of comparative 16S rRNA gene sequence analysis, belonged to the order Clostridiales in the phylum Firmicutes (low G+C gram-positive bacteria). Two of the strains were not involved in glycolate metabolism. The third, the glycolate-fermenting strain 19gly4 (DSM 11261), was related to members of the family Lachnospiraceae. The cells of strain 19gly4 were oval- to lemon-shaped, 0.85 microm long and 0.65 microm in diameter, occurring singly, in pairs, or in chains of up to 30 cells. Strain 19gly4 fermented glycolate or fumarate to acetate, succinate, and CO(2). Hydrogen was not formed, and strain 19gly4 was able to grow on glycolate in pure culture without any syntrophic hydrogen transfer and without the use of an external electron acceptor. There was no evidence for homoacetogenic metabolism. This bacterium therefore differs in metabolism from previously reported glycolate-utilising anaerobes.

Quick-start of full-scale anaerobic digestion (AD) using aeration

Energy Technology Data Exchange (ETDEWEB)

Lagerkvist, Anders, E-mail: al@ltu.se; Pelkonen, Markku; Wikström, Tommy

2015-04-15

Highlights: • A fast, and original, start up procedure for anaerobic digestors has been applied at full scale. • The development of a methanogenic culture has been documented using fluorescent in situ hybridization. • The technique can be widely applied. - Abstract: A conventional 1300 m{sup 3} continuously stirred anaerobic tank reactor at the city of Boden, north Sweden, which was receiving a feed of both sewage sludge and food waste, was put out of operation due to the build-up of a float phase. The reactor was emptied and cleaned. At start-up there was no methanogenic sludge available, so an unconventional start-up procedure was applied: The reactor was rapidly (8 days with 1200 kg of total solids (TS) added daily) filled with thickened, and slightly acidic sewage sludge, showing only slight methane generation, which was subsequently heated to 55 °C. Then compressed air was blown into the digester and within a month a fully functional methanogenic culture was established. The transfer from acidogenic to methanogenic conditions happened in about one week. As a start-up technique this is fast and cost efficient, it only requires the access of a compressor, electricity and a source of air. In total, about 16 tonnes of oxygen were used. It is proposed that this method may also be used as an operational amendment technique, should a reactor tend to acidify.

Quick-start of full-scale anaerobic digestion (AD) using aeration

International Nuclear Information System (INIS)

Lagerkvist, Anders; Pelkonen, Markku; Wikström, Tommy

2015-01-01

Highlights: • A fast, and original, start up procedure for anaerobic digestors has been applied at full scale. • The development of a methanogenic culture has been documented using fluorescent in situ hybridization. • The technique can be widely applied. - Abstract: A conventional 1300 m 3 continuously stirred anaerobic tank reactor at the city of Boden, north Sweden, which was receiving a feed of both sewage sludge and food waste, was put out of operation due to the build-up of a float phase. The reactor was emptied and cleaned. At start-up there was no methanogenic sludge available, so an unconventional start-up procedure was applied: The reactor was rapidly (8 days with 1200 kg of total solids (TS) added daily) filled with thickened, and slightly acidic sewage sludge, showing only slight methane generation, which was subsequently heated to 55 °C. Then compressed air was blown into the digester and within a month a fully functional methanogenic culture was established. The transfer from acidogenic to methanogenic conditions happened in about one week. As a start-up technique this is fast and cost efficient, it only requires the access of a compressor, electricity and a source of air. In total, about 16 tonnes of oxygen were used. It is proposed that this method may also be used as an operational amendment technique, should a reactor tend to acidify

Associative patterns among anaerobic fungi, methanogenic archaea, and bacterial communities in response to changes in diet and age in the rumen of dairy cows.

Science.gov (United States)

Kumar, Sanjay; Indugu, Nagaraju; Vecchiarelli, Bonnie; Pitta, Dipti W

2015-01-01

The rumen microbiome represents a complex microbial genetic web where bacteria, anaerobic rumen fungi (ARF), protozoa and archaea work in harmony contributing to the health and productivity of ruminants. We hypothesized that the rumen microbiome shifts as the dairy cow advances in lactations and these microbial changes may contribute to differences in productivity between primiparous (first lactation) and multiparous (≥second lactation) cows. To this end, we investigated shifts in the ruminal ARF and methanogenic communities in both primiparous (n = 5) and multiparous (n = 5) cows as they transitioned from a high forage to a high grain diet upon initiation of lactation. A total of 20 rumen samples were extracted for genomic DNA, amplified using archaeal and fungal specific primers, sequenced on a 454 platform and analyzed using QIIME. Community comparisons (Bray-Curtis index) revealed the effect of diet (P bacteria, ARF and archaea revealed syntrophic interactions both within and between microbial domains in response to change in diet as well as age of dairy cows. Notably, these interactions were numerous and complex in multiparous cows, supporting our hypothesis that the rumen microbiome also matures with age to sustain the growing metabolic needs of the host. This study provides a broader picture of the ARF and methanogenic populations in the rumen of dairy cows and their co-occurrence implicates specific relationships between different microbial domains in response to diet and age.

The Effects of Perchlorates on the Permafrost Methanogens: Implication for Autotrophic Life on Mars.

Science.gov (United States)

Shcherbakova, Viktoria; Oshurkova, Viktoria; Yoshimura, Yoshitaka

2015-09-09

The terrestrial permafrost represents a range of possible cryogenic extraterrestrial ecosystems on Earth-like planets without obvious surface ice, such as Mars. The autotrophic and chemolithotrophic psychrotolerant methanogens are more likely than aerobes to function as a model for life forms that may exist in frozen subsurface environments on Mars, which has no free oxygen, inaccessible organic matter, and extremely low amounts of unfrozen water. Our research on the genesis of methane, its content and distribution in permafrost horizons of different ages and origin demonstrated the presence of methane in permanently frozen fine-grained sediments. Earlier, we isolated and described four strains of methanogenic archaea of Methanobacterium and Methanosarcina genera from samples of Pliocene and Holocene permafrost from Eastern Siberia. In this paper we study the effect of sodium and magnesium perchlorates on growth of permafrost and nonpermafrost methanogens, and present evidence that permafrost hydogenotrophic methanogens are more resistant to the chaotropic agent found in Martian soil. In this paper we study the effect of sodium and magnesium perchlorates on the growth of permafrost and nonpermafrost methanogens, and present evidence that permafrost hydogenotrophic methanogens are more resistant to the chaotropic agent found in Martian soil. Furthermore, as shown in the studies strain M2(T) M. arcticum, probably can use perchlorate anion as an electron acceptor in anaerobic methane oxidation. Earth's subzero subsurface environments are the best approximation of environments on Mars, which is most likely to harbor methanogens; thus, a biochemical understanding of these pathways is expected to provide a basis for designing experiments to detect autotrophic methane-producing life forms on Mars.

The Effects of Perchlorates on the Permafrost Methanogens: Implication for Autotrophic Life on Mars

Directory of Open Access Journals (Sweden)

Viktoria Shcherbakova

2015-09-01

Full Text Available The terrestrial permafrost represents a range of possible cryogenic extraterrestrial ecosystems on Earth-like planets without obvious surface ice, such as Mars. The autotrophic and chemolithotrophic psychrotolerant methanogens are more likely than aerobes to function as a model for life forms that may exist in frozen subsurface environments on Mars, which has no free oxygen, inaccessible organic matter, and extremely low amounts of unfrozen water. Our research on the genesis of methane, its content and distribution in permafrost horizons of different ages and origin demonstrated the presence of methane in permanently frozen fine-grained sediments. Earlier, we isolated and described four strains of methanogenic archaea of Methanobacterium and Methanosarcina genera from samples of Pliocene and Holocene permafrost from Eastern Siberia. In this paper we study the effect of sodium and magnesium perchlorates on growth of permafrost and nonpermafrost methanogens, and present evidence that permafrost hydogenotrophic methanogens are more resistant to the chaotropic agent found in Martian soil. In this paper we study the effect of sodium and magnesium perchlorates on the growth of permafrost and nonpermafrost methanogens, and present evidence that permafrost hydogenotrophic methanogens are more resistant to the chaotropic agent found in Martian soil. Furthermore, as shown in the studies strain M2T M. arcticum, probably can use perchlorate anion as an electron acceptor in anaerobic methane oxidation. Earth’s subzero subsurface environments are the best approximation of environments on Mars, which is most likely to harbor methanogens; thus, a biochemical understanding of these pathways is expected to provide a basis for designing experiments to detect autotrophic methane-producing life forms on Mars.

Anaerobic bacterial quantitation of Yucca Mountain, Nevada DOE site samples

International Nuclear Information System (INIS)

Clarkson, W.W.; Krumholz, L.R.; Suflita, J.M.

1996-01-01

Anaerobic bacteria were studied from samples of excavated rock material as one phase of the overall Yucca Mountain site characterization effort. An indication of the abundance of important groups of anaerobic bacteria would enable inferences to be made regarding the natural history of the site and allow for more complete risk evaluation of the site as a nuclear repository. Six bacterial groups were investigated including anaerobic heterotrophs, acetogens, methanogens, sulfate-, nitrate-, and iron-reducing bacteria. The purpose of this portion of the study was to detect and quantify the aforementioned bacterial groups

Evolution of microorganisms in thermophilic-dry anaerobic digestion.

Science.gov (United States)

Montero, B; Garcia-Morales, J L; Sales, D; Solera, R

2008-05-01

Microbial population dynamics were studied during the start-up and stabilization periods in thermophilic-dry anaerobic digestion at lab-scale. The experimental protocol was defined to quantify Eubacteria and Archaea using Fluorescent in situ hybridization (FISH) in a continuously stirred tank reactor (CSTR), without recycling solids. The reactor was subjected to a programme of steady-state operation over a range of the retention times from 40 to 25 days, with an organic loading rate between 4.42 and 7.50 kg volatile solid/m3/day. Changes in microbial concentrations were linked to traditional performance parameters such as biogas production and VS removal. The relations of Eubacteria:Archaea and H2-utilising methanogens:acetate-utilising methanogens were 88:12 and 11:1, respectively, during start-up stage. Hydrogenotrophic methanogens, although important in the initial phase of the reactor start-up, were displaced by acetoclastic methanogens at steady-state, thus their relation were 7:32, respectively. The methane yield coefficient, the methane content in the biogas and VS removal were stabilized around 0.30 LCH4/gCOD, 50% and 80%, respectively. Methanogenic population correlated well with performance measurements.

Biodegradation and reversible inhibitory impact of sulfamethoxazole on the utilization of volatile fatty acids during anaerobic treatment of pharmaceutical industry wastewater

International Nuclear Information System (INIS)

Cetecioglu, Zeynep; Ince, Bahar; Gros, Meritxell; Rodriguez-Mozaz, Sara; Barceló, Damia; Ince, Orhan; Orhon, Derin

2015-01-01

This study evaluated the chronic impact and biodegradability of sulfamethoxazole under anaerobic conditions. For this purpose, a lab-scale anaerobic sequencing batch reactor was operated in a sequence of different phases with gradually increasing sulfamethoxazole doses of 1 to 45 mg/L. Conventional parameters, such as COD, VFA, and methane generation, were monitored with corresponding antimicrobial concentrations in the reactor and the methanogenic activity of the sludge. The results revealed that anaerobic treatment was suitable for pharmaceutical industry wastewater with concentrations of up to 40 mg/L of sulfamethoxazole. Higher levels exerted toxic effects on the microbial community under anaerobic conditions, causing the inhibition of substrate/COD utilization and biogas generation and leading to a total collapse of the reactor. The adverse long-term impact was quite variable for fermentative bacteria and methanogenic achaea fractions of the microbial community based on changes inflicted on the composition of the residual organic substrate and mRNA expression of the key enzymes. - Highlights: • Chronic impact of sulfamethoxazole was lethal at 45 mg/L on the microbial community. • Sulfamethoxazole was highly biodegradable under anaerobic conditions. • While the COD removal stopped, the sorption of sulfamethoxazole into the sludge increased. • Sulfamethoxazole has a reversible inhibitory effect on acetoclastic methanogens

Biodegradation and reversible inhibitory impact of sulfamethoxazole on the utilization of volatile fatty acids during anaerobic treatment of pharmaceutical industry wastewater

Energy Technology Data Exchange (ETDEWEB)

Cetecioglu, Zeynep, E-mail: cetecioglu@itu.edu.tr [Istanbul Technical University, Environmental Engineering Department, 34469 Maslak, Istanbul (Turkey); Catalan Institute for Water Research (ICRA), Emili Grahit 101, 17003 Girona (Spain); Ince, Bahar [Bogazici University, Institute of Environmental Sciences, Rumelihisarustu - Bebek, 34342 Istanbul (Turkey); Gros, Meritxell; Rodriguez-Mozaz, Sara; Barceló, Damia [Catalan Institute for Water Research (ICRA), Emili Grahit 101, 17003 Girona (Spain); Ince, Orhan; Orhon, Derin [Istanbul Technical University, Environmental Engineering Department, 34469 Maslak, Istanbul (Turkey)

2015-12-01

This study evaluated the chronic impact and biodegradability of sulfamethoxazole under anaerobic conditions. For this purpose, a lab-scale anaerobic sequencing batch reactor was operated in a sequence of different phases with gradually increasing sulfamethoxazole doses of 1 to 45 mg/L. Conventional parameters, such as COD, VFA, and methane generation, were monitored with corresponding antimicrobial concentrations in the reactor and the methanogenic activity of the sludge. The results revealed that anaerobic treatment was suitable for pharmaceutical industry wastewater with concentrations of up to 40 mg/L of sulfamethoxazole. Higher levels exerted toxic effects on the microbial community under anaerobic conditions, causing the inhibition of substrate/COD utilization and biogas generation and leading to a total collapse of the reactor. The adverse long-term impact was quite variable for fermentative bacteria and methanogenic achaea fractions of the microbial community based on changes inflicted on the composition of the residual organic substrate and mRNA expression of the key enzymes. - Highlights: • Chronic impact of sulfamethoxazole was lethal at 45 mg/L on the microbial community. • Sulfamethoxazole was highly biodegradable under anaerobic conditions. • While the COD removal stopped, the sorption of sulfamethoxazole into the sludge increased. • Sulfamethoxazole has a reversible inhibitory effect on acetoclastic methanogens.

Archaea and Bacteria Acclimate to High Total Ammonia in a Methanogenic Reactor Treating Swine Waste

Directory of Open Access Journals (Sweden)

Sofia Esquivel-Elizondo

2016-01-01

Full Text Available Inhibition by ammonium at concentrations above 1000 mgN/L is known to harm the methanogenesis phase of anaerobic digestion. We anaerobically digested swine waste and achieved steady state COD-removal efficiency of around 52% with no fatty-acid or H2 accumulation. As the anaerobic microbial community adapted to the gradual increase of total ammonia-N (NH3-N from 890±295 to 2040±30 mg/L, the Bacterial and Archaeal communities became less diverse. Phylotypes most closely related to hydrogenotrophic Methanoculleus (36.4% and Methanobrevibacter (11.6%, along with acetoclastic Methanosaeta (29.3%, became the most abundant Archaeal sequences during acclimation. This was accompanied by a sharp increase in the relative abundances of phylotypes most closely related to acetogens and fatty-acid producers (Clostridium, Coprococcus, and Sphaerochaeta and syntrophic fatty-acid Bacteria (Syntrophomonas, Clostridium, Clostridiaceae species, and Cloacamonaceae species that have metabolic capabilities for butyrate and propionate fermentation, as well as for reverse acetogenesis. Our results provide evidence countering a prevailing theory that acetoclastic methanogens are selectively inhibited when the total ammonia-N concentration is greater than ~1000 mgN/L. Instead, acetoclastic and hydrogenotrophic methanogens coexisted in the presence of total ammonia-N of ~2000 mgN/L by establishing syntrophic relationships with fatty-acid fermenters, as well as homoacetogens able to carry out forward and reverse acetogenesis.

Comparison of Rumen and Manure Microbiomes and Implications for the Inoculation of Anaerobic Digesters.

Science.gov (United States)

Ozbayram, Emine Gozde; Ince, Orhan; Ince, Bahar; Harms, Hauke; Kleinsteuber, Sabine

2018-02-14

Cattle manure is frequently used as an inoculum for the start-up of agricultural biogas plants or as a co-substrate in the anaerobic digestion of lignocellulosic feedstock. Ruminal microbiota are considered to be effective plant fiber degraders, but the microbes contained in manure do not necessarily reflect the rumen microbiome. The aim of this study was to compare the microbial community composition of cow rumen and manure with respect to plant fiber-digesting microbes. Bacterial and methanogenic communities of rumen and manure samples were examined by 454 amplicon sequencing of bacterial 16S rRNA genes and mcrA genes, respectively. Rumen fluid samples were dominated by Prevotellaceae (29%), whereas Ruminococcaceae was the most abundant family in the manure samples (31%). Fibrobacteraceae (12%) and Bacteroidaceae (13%) were the second most abundant families in rumen fluid and manure, respectively. The high abundances of fiber-degrading bacteria belonging to Prevotellaceae and Fibrobacteraceae might explain the better performance of anaerobic digesters inoculated with rumen fluid. Members of the genus Methanobrevibacter were the predominant methanogens in the rumen fluid, whereas methanogenic communities of the manure samples were dominated by the candidate genus Methanoplasma . Our results suggest that inoculation or bioaugmentation with fiber-digesting rumen microbiota can enhance the anaerobic digestion of lignocellulosic biomass.

Comparison of Rumen and Manure Microbiomes and Implications for the Inoculation of Anaerobic Digesters

Directory of Open Access Journals (Sweden)

Emine Gozde Ozbayram

2018-02-01

Full Text Available Cattle manure is frequently used as an inoculum for the start-up of agricultural biogas plants or as a co-substrate in the anaerobic digestion of lignocellulosic feedstock. Ruminal microbiota are considered to be effective plant fiber degraders, but the microbes contained in manure do not necessarily reflect the rumen microbiome. The aim of this study was to compare the microbial community composition of cow rumen and manure with respect to plant fiber-digesting microbes. Bacterial and methanogenic communities of rumen and manure samples were examined by 454 amplicon sequencing of bacterial 16S rRNA genes and mcrA genes, respectively. Rumen fluid samples were dominated by Prevotellaceae (29%, whereas Ruminococcaceae was the most abundant family in the manure samples (31%. Fibrobacteraceae (12% and Bacteroidaceae (13% were the second most abundant families in rumen fluid and manure, respectively. The high abundances of fiber-degrading bacteria belonging to Prevotellaceae and Fibrobacteraceae might explain the better performance of anaerobic digesters inoculated with rumen fluid. Members of the genus Methanobrevibacter were the predominant methanogens in the rumen fluid, whereas methanogenic communities of the manure samples were dominated by the candidate genus Methanoplasma. Our results suggest that inoculation or bioaugmentation with fiber-digesting rumen microbiota can enhance the anaerobic digestion of lignocellulosic biomass.

A characterization of anaerobic colonization and associated mucosal adaptations in the undiseased ileal pouch.

LENUS (Irish Health Repository)

Smith, F M

2012-02-03

INTRODUCTION: The resolution of pouchitis with metronidazole points to an anaerobic aetiology. Pouchitis is mainly seen in patients with ulcerative colitis pouches (UCP). We have recently found that sulphate reducing bacteria (SRB), a species of strict anaerobe, colonize UCP exclusively. Herein, we aimed to correlate levels of different bacterial species (including SRB) with mucosal inflammation and morphology. METHODS: Following ethical approval, fresh faecal samples and mucosal biopsies were taken from 9 patients with UCP and 5 patients with familial adenomatous polyposis pouches (FAPP). For the purposes of comparison, faecal samples and mucosal biopsies were also taken from the stomas of 7 of the 9 patients with UC (UCS). Colonization by four types of strict anaerobes (SRB, Clostridium perfringens, Bifidobacteria and Bacteroides) as well as by three types of facultative anaerobes (Enterococci, Coliforms and Lactobacilli) was evaluated. Inflammatory scores and mucosal morphology were assessed histologically in a blinded fashion by a pathologist. RESULTS: In general, strict anaerobes predominated over facultative in the UCP (P = 0.041). SRB were present in UCP exclusively. Even after exclusion of SRB from total bacterial counts, strict anaerobes still predominated. In the UCS, facultative anaerobes predominated. Strict and facultative anaerobes were present at similar levels in the FAPP. Enterococci were present at significantly reduced levels in the UCP when compared with the UCS (P = 0.031). When levels of SRB and other anaerobic species were individually correlated with mucosal inflammation and morphology, no trends were observed. CONCLUSION: We have previously identified that SRB exclusively colonize UCP. In addition we have now identified a novel increase in the strict\\/facultative anaerobic ratio within the UCP compared to UCS. These stark differences in bacterial colonization, however, appear to have limited impact on mucosal inflammation or morphology.

Performance of thermophilic anaerobic digesters using inoculum mixes with enhanced methanogenic diversity

KAUST Repository

Ghanimeh, Sophia; El-Fadel, Mutasem; Saikaly, Pascal

2017-01-01

Reportedly, various mixes of seeds were quasi-randomly selected to startup anaerobic digesters. In contrast, this study examines the impact of inoculating thermophilic anaerobic digesters with a designed mix of non-acclimated seeds based

Reductive dechlorination of hexachlorocyclohexane (HCH) isomers in soil under anaerobic conditions

NARCIS (Netherlands)

Middeldorp, P.J.M.; Doesburg, van W.C.J.; Schraa, G.; Stams, A.J.M.

2005-01-01

The biological anaerobic reductive dechlorination of -hexachlorocyclohexane under methanogenic conditions was tested in a number of contaminated soil samples from two locations in the Netherlands. Soils from a heavily polluted location showed rapid dechlorination of -hexachlorocyclohexane to benzene

Microbial community shifts in a farm-scale anaerobic digester treating swine waste: Correlations between bacteria communities associated with hydrogenotrophic methanogens and environmental conditions.

Science.gov (United States)

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

2017-12-01

Microbial community structure in a farm-scale anaerobic digester treating swine manure was investigated during three process events: 1) prolonged starvation, and changes of 2) operating temperature (between meso- and thermophilic) and 3) hydraulic retention time (HRT). Except during the initial period, the digester was dominated by hydrogenotrophic methanogens (HMs). The bacterial community structure significantly shifted with operating temperature and HRT but not with long-term starvation. Clostridiales (26.5-54.4%) and Bacteroidales (2.5-13.7%) became dominant orders in the digester during the period of HM dominance. Abundance of diverse meso- and thermophilic bacteria increased during the same period; many of these species may be H 2 producers, and/or syntrophic acetate oxidizers. Some of these species showed positive correlations with [NH 4 + -N] (panaerobic digesters treating swine manure that contains high ammonia content. Copyright © 2017 Elsevier B.V. All rights reserved.

A comparative study of leachate quality and biogas generation in simulated anaerobic and hybrid bioreactors

Energy Technology Data Exchange (ETDEWEB)

Xu, Qiyong; Tian, Ying; Wang, Shen; Ko, Jae Hac, E-mail: jaehacko@pkusz.edu.cn

2015-07-15

Highlights: • Temporary aeration shortened the initial acid inhibition phase for methanogens. • COD decreased faster in the hybrid bioreactor than that in the anaerobic control. • Methane generations from hybrid bioreactors were 133.4 L/kg{sub vs} and 113.2 L/kg{sub vs}. • MSW settlement increased with increasing the frequency of intermittent aeration. - Abstract: Research has been conducted to compare leachate characterization and biogas generation in simulated anaerobic and hybrid bioreactor landfills with typical Chinese municipal solid waste (MSW). Three laboratory-scale reactors, an anaerobic (A1) and two hybrid bioreactors (C1 and C2), were constructed and operated for about 10 months. The hybrid bioreactors were operated in an aerobic–anaerobic mode with different aeration frequencies by providing air into the upper layer of waste. Results showed that the temporary aeration into the upper layer aided methane generation by shortening the initial acidogenic phase because of volatile fatty acids (VFAs) reduction and pH increase. Chemical oxygen demand (COD) decreased faster in the hybrid bioreactors, but the concentrations of ammonia–nitrogen in the hybrid bioreactors were greater than those in the anaerobic control. Methanogenic conditions were established within 75 d and 60 d in C1 and C2, respectively. However, high aeration frequency led to the consumption of organic matters by aerobic degradation and resulted in reducing accumulative methane volume. The temporary aeration enhanced waste settlement and the settlement increased with increasing the frequency of aeration. Methane production was inhibited in the anaerobic control; however, the total methane generations from hybrid bioreactors were 133.4 L/kg{sub vs} and 113.2 L/kg{sub vs}. As for MSW with high content of food waste, leachate recirculation right after aeration stopped was not recommended due to VFA inhibition for methanogens.

Correlation between microbial community and granule conductivity in anaerobic bioreactors for brewery wastewater treatment

DEFF Research Database (Denmark)

Shrestha, Pravin; Malvankar, Nikhil S.; Werner, Jeffrey

2014-01-01

Prior investigation of an upflow anaerobic sludge blanket (UASB) reactor treating brewery wastes suggested that direct interspecies electron transfer (DIET) significantly contributed to interspecies electron transfer to methanogens. To investigate DIET in granules further, the electrical conducti......Prior investigation of an upflow anaerobic sludge blanket (UASB) reactor treating brewery wastes suggested that direct interspecies electron transfer (DIET) significantly contributed to interspecies electron transfer to methanogens. To investigate DIET in granules further, the electrical...... conductivity and bacterial community composition of granules in fourteen samples from four different UASB reactors treating brewery wastes were investigated. All of the UASB granules were electrically conductive whereas control granules from ANAMMOX (ANaerobic AMMonium OXidation) reactors and microbial...... granules from an aerobic bioreactor designed for phosphate removal were not. There was a moderate correlation (r = 0.67) between the abundance of Geobacter species in the UASB granules and granule conductivity, suggesting that Geobacter contributed to granule conductivity. These results, coupled...

Azo dye reduction by mesophilic and thermophilic anaerobic consortia

NARCIS (Netherlands)

Santos, dos A.B.; Madrid, de M.P.; Stams, A.J.M.; Lier, van J.B.; Cervantes, F.J.

2005-01-01

The reduction of the azo dye model compounds Reactive Red 2 (RR2) and Reactive Orange 14 (RO14) by mesophilic (30 C) and thermophilic (55 C) anaerobic consortia was studied in batch assays. The contribution of fermentative and methanogenic microorganisms in both temperatures was evaluated in the

Biogeochemistry of anaerobic crude oil biodegradation

Science.gov (United States)

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

2010-05-01

Anaerobic degradation of crude oil and petroleum hydrocarbons is widely recognized as a globally significant process both in the formation of the world's vast heavy oil deposits and for the dissipation of hydrocarbon pollution in anoxic contaminated environments. Comparative analysis of crude oil biodegradation under methanogenic and sulfate-reducing conditions has revealed differences not only in the patterns of compound class removal but also in the microbial communities responsible. Under methanogenic conditions syntrophic associations dominated by bacteria from the Syntropheaceae are prevalent and these are likely key players in the initial anaerobic degradation of crude oil alkanes to intermediates such as hydrogen and acetate. Syntrophic acetate oxidation plays an important role in these systems and often results in methanogenesis dominated by CO2 reduction by members of the Methanomicrobiales. By contrast the bacterial communities from sulfate-reducing crude oil-degrading systems were more diverse and no single taxon dominated the oil-degrading sulfate-reducing systems. All five proteobacterial subdivisions were represented with Delta- and Gammaproteobacteria being detected most consistently. In sediments which were pasteurized hydrocarbon degradation continued at a relatively low rate. Nevertheless, alkylsuccinates characteristic of anaerobic hydrocarbon degradation accumulated to high concentrations. This suggested that the sediments harbour heat resistant, possibly spore-forming alkane degrading sulfate-reducers. This is particularly interesting since it has been proposed recently, that spore-forming sulfate-reducing bacteria found in cold arctic sediments may have originated from seepage of geofluids from deep subsurface hydrocarbon reservoirs.

Impact of fulvic acids on bio-methanogenic treatment of municipal solid waste incineration leachate.

Science.gov (United States)

Dang, Yan; Lei, Yuqing; Liu, Zhao; Xue, Yiting; Sun, Dezhi; Wang, Li-Ying; Holmes, Dawn E

2016-12-01

A considerable amount of leachate with high fulvic acid (FA) content is generated during the municipal solid waste (MSW) incineration process. This incineration leachate is usually processed by downstream bio-methanogenic treatment. However, few studies have examined the impact that these compounds have on methanogenesis and how they are degraded and transformed during the treatment process. In this study, a laboratory-scale expanded granular sludge bed (EGSB) reactor was operated with MSW incineration leachate containing various concentrations of FA (1500 mg/L to 8000 mg/L) provided as the influent. We found that FA degradation rates decreased from 86% to 72% when FA concentrations in the reactor were increased, and that molecular size, level of humification and aromatization of the residual FA macromolecules all increased after bio-methanogenic treatment. Increasing FA influent concentrations also inhibited growth of hydrogenotrophic methanogens from the genus Methanobacterium and syntrophic bacteria from the genus Syntrophomonas, which resulted in a decrease in methane production and a concomitant increase in CO 2 content in the biogas. Sequences most similar to species from the genus Anaerolinea went up as FA concentrations increased. Bacteria from this genus are capable of extracellular electron transfer and may be using FA as an electron acceptor for growth or as a shuttle for syntrophic exchange with other microorganisms in the reactor. In order to determine whether FA could serve as an electron shuttle to promote syntrophy in an anaerobic digester, co-cultures of Geobacter metallireducens and G. sulfurreducens were grown in the presence of FA from raw leachate or from residual bioreactor effluent. While raw FA stimulated electron transfer between these two bacteria, residual FA did not have any electron shuttling abilities, indicating that FA underwent a significant transformation during the bio-methanogenic treatment process. These results are

Converting mesophilic upflow sludge blanket (UASB) reactors to thermophilic by applying axenic methanogenic culture bioaugmentation

DEFF Research Database (Denmark)

Zhu, Xinyu; Treu, Laura; Kougias, Panagiotis G.

2018-01-01

on the microbial consortium. The adaptation of microbial community to a new environment or condition can be accelerated by a process known as “bioaugmentation” or “microbial community manipulation”, during which exogenous microorganisms harbouring specific metabolic activities are introduced to the reactor....... The aim of the current study was to rapidly convert the operational temperature of up-flow anaerobic sludge blanket (UASB) reactors from mesophilic to thermophilic conditions by applying microbial community manipulation techniques. Three different bioaugmentation strategies were compared and it was proven...... that the injection of axenic methanogenic culture was the most efficient approach leading to improved biomethanation process with 40% higher methane production rate compared to the control reactor. Microbial community analyses revealed that during bioaugmentation, the exogenous hydrogenotrophic methanogen could...

Anaerobic biodegradation of (emerging) organic contaminants in the aquatic environment.

Science.gov (United States)

Ghattas, Ann-Kathrin; Fischer, Ferdinand; Wick, Arne; Ternes, Thomas A

2017-06-01

Although strictly anaerobic conditions prevail in several environmental compartments, up to now, biodegradation studies with emerging organic contaminants (EOCs), such as pharmaceuticals and personal care products, have mainly focused on aerobic conditions. One of the reasons probably is the assumption that the aerobic degradation is more energetically favorable than degradation under strictly anaerobic conditions. Certain aerobically recalcitrant contaminants, however, are biodegraded under strictly anaerobic conditions and little is known about the organisms and enzymatic processes involved in their degradation. This review provides a comprehensive survey of characteristic anaerobic biotransformation reactions for a variety of well-studied, structurally rather simple contaminants (SMOCs) bearing one or a few different functional groups/structural moieties. Furthermore it summarizes anaerobic degradation studies of more complex contaminants with several functional groups (CMCs), in soil, sediment and wastewater treatment. While strictly anaerobic conditions are able to promote the transformation of several aerobically persistent contaminants, the variety of observed reactions is limited, with reductive dehalogenations and the cleavage of ether bonds being the most prevalent. Thus, it becomes clear that the transferability of degradation mechanisms deduced from culture studies of SMOCs to predict the degradation of CMCs, such as EOCs, in environmental matrices is hampered due the more complex chemical structure bearing different functional groups, different environmental conditions (e.g. matrix, redox, pH), the microbial community (e.g. adaptation, competition) and the low concentrations typical for EOCs. Copyright © 2017 The Authors. Published by Elsevier Ltd.. All rights reserved.

Microbiological evaluation of a raw mud coming from the treatment of residual waters plant of the Municipality of The Retiro and of the same mud acclimatized to anaerobic conditions

International Nuclear Information System (INIS)

Naranjo P, Fernando; Gonzalez D, Maria Elena; Molina P, Francisco

2001-01-01

The subject of this study is the acclimatization of aerobic sludge (activated sludge) up to anaerobic sludge conditions, the activated sludge from a domestic sewage treatment plant; which is located in El Retiro town east of Antioquia. Load potential of brown sugar was applied in the batch reactor 280-2.000 mg COD/I. In the acclimatization process, both sludge were assessed the groups of bacteria involved in the anaerobic digestion as total MPN count and oxygen sensibility as two methods; furthermore, and also Volatile Suspend Solids (VSS), methanogenic specific activity where acetic and formic as substrates were used. The content of VSS was recognize as a good parameter to assess both activated sludge; besides, as a result of methanogenic, specific activity show an important increase in the acclimated sludge with formic; which is corroborated with an increase inc the group of hydrogenophilic methanogenic bacteria. At the end, the activated sludge from activated treatment plant, was identified as a potential inocula in anaerobic process, because there are some quantities of methanogenic bacteria

Physiologically anaerobic microorganisms of the deep subsurface. Progress report, June 1, 1991--May 31, 1992

Energy Technology Data Exchange (ETDEWEB)

Stevens, S.E. Jr.; Chung, K.T.

1992-06-01

A variety of different media were used to isolate facultatively (FAB) and obligately anaerobic bacteria (OAB). These bacteria were isolated from core subsamples obtained from boreholes at the Idaho National Engineering Lab. (INEL) or at the Hanford Lab. (Yakima). Core material was sampled at various depths to 600 feet below the surface. All core samples with culturable bacteria contained at least FAB making thisthe most common physiological type of anaerobic bacteria present in the deep subsurface at these two sites. INEL core samples are characterized by isolates of both FAB and OAB. No isolates of acetogenic, methanogenic, or sulfate reducing bacteria were obtained. Yakima core samples are characterized by a marked predominance of FAB in comparison to OAB. In addition, isolates of acetogenic, methanogenic, and sulfate reducing bacteria were obtained. The Yakima site has the potential for complete anaerobic mineralization of organic compounds whereas this potential appears to be lacking at INEL.

A field experiment for the anaerobic biotransformation of aromatic hydrocarbon compounds at Seal Beach, California

International Nuclear Information System (INIS)

Reinhard, M.; Wills, L.E.; Ball, H.A.; Harmon, T.

1991-01-01

Biotransformation of aromatic hydrocarbons under anaerobic conditions is of interest because dissolved oxygen is rapidly consumed in groundwater contaminant plumes of hydrocarbon fuel. Anaerobic biotransformation of aromatic hydrocarbons has been demonstrated under different redox regimes including nitrate-reducing iron-reducing and fermentative-methanogenic conditions. Recently, laboratory evidence has been obtained for the degradation of alkylbenzenes including toluene under sulfate-reducing conditions. The long-term objective of this study is to determine transformation rates under the conditions of the Seal Beach site, and second to explore the feasibility of inducing nitrate- and sulfate-reducing conditions and fermentative-methanogenic conditions in field bioreactors. Both laboratory studies and field studies in bioreactors are being conducted. This paper reports on the experimental design of the bioreactors and initial results

Biologically Produced Methane as a Renewable Energy Source.

Science.gov (United States)

Holmes, D E; Smith, J A

2016-01-01

Methanogens are a unique group of strictly anaerobic archaea that are more metabolically diverse than previously thought. Traditionally, it was thought that methanogens could only generate methane by coupling the oxidation of products formed by fermentative bacteria with the reduction of CO 2 . However, it has recently been observed that many methanogens can also use electrons extruded from metal-respiring bacteria, biocathodes, or insoluble electron shuttles as energy sources. Methanogens are found in both human-made and natural environments and are responsible for the production of ∼71% of the global atmospheric methane. Their habitats range from the human digestive tract to hydrothermal vents. Although biologically produced methane can negatively impact the environment if released into the atmosphere, when captured, it can serve as a potent fuel source. The anaerobic digestion of wastes such as animal manure, human sewage, or food waste produces biogas which is composed of ∼60% methane. Methane from biogas can be cleaned to yield purified methane (biomethane) that can be readily incorporated into natural gas pipelines making it a promising renewable energy source. Conventional anaerobic digestion is limited by long retention times, low organics removal efficiencies, and low biogas production rates. Therefore, many studies are being conducted to improve the anaerobic digestion process. Researchers have found that addition of conductive materials and/or electrically active cathodes to anaerobic digesters can stimulate the digestion process and increase methane content of biogas. It is hoped that optimization of anaerobic digesters will make biogas more readily accessible to the average person. Copyright © 2016 Elsevier Inc. All rights reserved.

Anaerobic biodegradation of cyanide under methanogenic conditions.

Science.gov (United States)

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

Gelria glutamica gen. nov., sp. a thermophilic oligately syntrophic glutamate-degrading anaerobe

NARCIS (Netherlands)

Plugge, C.M.; Balk, M.; Zoetendal, E.G.; Stams, A.J.M.

2002-01-01

A novel anaerobic, Gram-positive, thermophilic, spore-forming, obligately syntrophic, glutamate-degrading bacterium, strain TGO(T), was isolated from a propionate-oxidizing methanogenic enrichment culture. The axenic culture was obtained by growing the bacterium on pyruvate. Cells were rod-shaped

Inhibition of the anaerobic digestion process by linear alkylbenzene sulfonates

DEFF Research Database (Denmark)

Gavala, Hariklia N.; Ahring, Birgitte Kiær

2002-01-01

Linear Alkylbenzene Sulfonates (LAS) are the most widely used synthetic anionic surfactants. They are anthropogenic, toxic compounds and are found in the primary sludge generated in municipal wastewater treatment plants. Primary sludge is usually stabilized anaerobically and therefore it is impor......Linear Alkylbenzene Sulfonates (LAS) are the most widely used synthetic anionic surfactants. They are anthropogenic, toxic compounds and are found in the primary sludge generated in municipal wastewater treatment plants. Primary sludge is usually stabilized anaerobically and therefore...... it is important to investigate the effect of these xenobiotic compounds on an anaerobic environment. The inhibitory effect of Linear Alkylbenzene Sulfonates (LAS) on the acetogenic and methanogenic step of the anaerobic digestion process was studied. LAS inhibit both acetogenesis from propionate...

Bacterial ecology of abattoir wastewater treated by an anaerobic digestor

Directory of Open Access Journals (Sweden)

Linda Jabari

2016-03-01

Full Text Available Abstract Wastewater from an anaerobic treatment plant at a slaughterhouse was analysed to determine the bacterial biodiversity present. Molecular analysis of the anaerobic sludge obtained from the treatment plant showed significant diversity, as 27 different phyla were identified. Firmicutes, Proteobacteria, Bacteroidetes, Thermotogae, Euryarchaeota (methanogens, and msbl6 (candidate division were the dominant phyla of the anaerobic treatment plant and represented 21.7%, 18.5%, 11.5%, 9.4%, 8.9%, and 8.8% of the total bacteria identified, respectively. The dominant bacteria isolated were Clostridium, Bacteroides, Desulfobulbus, Desulfomicrobium, Desulfovibrio and Desulfotomaculum. Our results revealed the presence of new species, genera and families of microorganisms. The most interesting strains were characterised. Three new bacteria involved in anaerobic digestion of abattoir wastewater were published.

Confocal Raman microspectroscopy reveals a convergence of the chemical composition in methanogenic archaea from a Siberian permafrost-affected soil.

Science.gov (United States)

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. © FEMS 2015. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

Problematic effects of antibiotics on anaerobic treatment of swine wastewater.

Science.gov (United States)

Cheng, D L; Ngo, H H; Guo, W S; Chang, S W; Nguyen, D D; Kumar, S Mathava; Du, B; Wei, Q; Wei, D

2018-05-04

Swine wastewaters with high levels of organic pollutants and antibiotics have become serious environmental concerns. Anaerobic technology is a feasible option for swine wastewater treatment due to its advantage in low costs and bioenergy production. However, antibiotics in swine wastewater have problematic effects on micro-organisms, and the stability and performance of anaerobic processes. Thus, this paper critically reviews impacts of antibiotics on pH, COD removal efficiencies, biogas and methane productions as well as the accumulation of volatile fatty acids (VFAs) in the anaerobic processes. Meanwhile, impacts on the structure of bacteria and methanogens in anaerobic processes are also discussed comprehensively. Furthermore, to better understand the effect of antibiotics on anaerobic processes, detailed information about antimicrobial mechanisms of antibiotics and microbial functions in anaerobic processes is also summarized. Future research on deeper knowledge of the effect of antibiotics on anaerobic processes are suggested to reduce their adverse environmental impacts. Copyright © 2018 Elsevier Ltd. All rights reserved.

Anaerobic digestion of molasses by means of a vibrating and non-vibrating submerged anaerobic membrane bioreactor

International Nuclear Information System (INIS)

De Vrieze, Jo; Hennebel, Tom; Van den Brande, Jens; Bilad, Ro'il M.; Bruton, Thomas A.; Vankelecom, Ivo F.J.; Verstraete, Willy; Boon, Nico

2014-01-01

Bio-refineries produce large volumes of waste streams with high organic content, which are potentially interesting for further processing. Anaerobic digestion (AD) can be a key technology for treatment of these sidestreams, such as molasses. However, the high concentration of salts in molasses can cause inhibition of methanogenesis. In this research, concentrated and diluted molasses were subjected to biomethanation in two types of submerged anaerobic membrane bioreactors (AnMBRs): one with biogas recirculation and one with a vibrating membrane. Both reactors were compared in terms of methane production and membrane fouling. Biogas recirculation seemed to be a good way to avoid membrane fouling, while the trans membrane pressures in the vibrating MBR increased over time, due to cake layer formation and the absence of a mixing system. Stable methane production, up to 2.05 L L −1  d −1 and a concomitant COD removal of 94.4%, was obtained only when diluted molasses were used, since concentrated molasses caused a decrease in methane production and an increase in volatile fatty acids (VFA), indicating an inhibiting effect of concentrated molasses on AD. Real-time PCR results revealed a clear dominance of Methanosaetaceae over Methanosarcinaceae as the main acetoclastic methanogens in both AnMBRs. - Highlights: • An anaerobic membrane bioreactor (AnMBR) can be used to digest diluted molasses. • Biogas recirculation is a good way to avoid fouling in an AnMBR. • Trans membrane pressures in AnMBR with vibrating membrane increased over time. • Methanosaeta sp. were the dominant acetoclastic methanogens

Anaerobic digestion of solid material

DEFF Research Database (Denmark)

Vavilin, V.A.; Lokshina, L.Y.; Flotats, X.

2007-01-01

A new multidimensional (3 and 2D) anaerobic digestion model for cylindrical reactor with non-uniform influent concentration distributions was developed to study the way in which mixing intensity affects the efficiency of continuous-flow anaerobic digestion. Batch experiments reported and simulated...... earlier by Vavilin and Angelidaki (2005) were used to modernize a kinetic scheme and to obtain the corresponding kinetic coefficients. In the new models, hydrolytic microorganisms were included using Contois kinetics for the hydrolysis/acidogenesis degradation of municipal solid waste (MSW). Monod...... kinetics was applied for description of methanogenesis. Both hydrolytic and methanogenic microorganisms were assumed to be inhibited by high volatile fatty acids (VFA) concentration. According to the new distributed models, the mixing level reduction expressed by increasing dimensionless Peclet number may...

Anaerobic biodegradability and treatment of grey water in upflow anaerobic sludge blanket (UASB) reactor.

Science.gov (United States)

Elmitwalli, Tarek A; Otterpohl, Ralf

2007-03-01

Feasibility of grey water treatment in an upflow anaerobic sludge blanket (UASB) reactor operated at different hydraulic retention time (HRT) of 16, 10 and 6h and controlled temperature of 30 degrees C was investigated. Moreover, the maximum anaerobic biodegradability without inoculum addition and maximum removal of chemical oxygen demand (COD) fractions in grey water were determined in batch experiments. High values of maximum anaerobic biodegradability (76%) and maximum COD removal in the UASB reactor (84%) were achieved. The results showed that the colloidal COD had the highest maximum anaerobic biodegradability (86%) and the suspended and dissolved COD had similar maximum anaerobic biodegradability of 70%. Furthermore, the results of the UASB reactor demonstrated that a total COD removal of 52-64% was obtained at HRT between 6 and 16 h. The UASB reactor removed 22-30% and 15-21% of total nitrogen and total phosphorous in the grey water, respectively, mainly due to the removal of particulate nutrients. The characteristics of the sludge in the UASB reactor confirmed that the reactor had a stable performance. The minimum sludge residence time and the maximum specific methanogenic activity of the sludge ranged between 27 and 93 days and 0.18 and 0.28 kg COD/(kg VS d).

[Identification and susceptibility to antimicrobial agents of strictly anaerobic bacteria isolated from hospitalized patients].

Science.gov (United States)

Kot, Katarzyna; Rokosz, Alicja; Sawicka-Grzelak, Anna; łuczak, MirosŁaw

2002-01-01

The aim of this study was to identify anaerobic strains isolated in 2001 from clinical specimens obtained from patients of Warsaw hospital and to evaluate a susceptibility of these strains to antimicrobial agents. In 2001 two hundred and twenty five clinical strains of obligate anaerobes were cultured, which were identified in the automatic ATB system (bioMérieux, France) using biochemical tests API 20 A. Drug-susceptibility of strains was determined also in ATB system with the use of ATB ANA strips. C. difficile strains were isolated on selective CCCA medium. Toxins A/B of C. difficile directly in stool specimens were detected by means of ELISA test (TechLab, USA). Fifty four strains of Gram-negative anaerobes (B. fragilis strains dominated) and 171 strains of Gram-positive anaerobes (the greatest number of strains belonged to genus Peptostreptococcus) were cultured from clinical specimens. In the cases of antibiotic-associated diarrhea 28 C. difficile strains were isolated and C. difficile toxins A/B were detected in 39 stool samples. The most active in vitro antimicrobials against Gram-negative anaerobes were metronidazole, imipenem, ticarcillin combined with clavulanic acid and piperacillin with tazobactam. Gram-positive, clinical strains of anaerobes were the most susceptible in vitro to beta-lactam antibiotics combined with beta-lactamase inhibitors (amoxicillin/clavulanate, piperacillin/tazobactam, ticarcillin/clavulanate) and imipenem.

Status on Science and Application of Thermophilic Anaerobic Digestion

DEFF Research Database (Denmark)

Ahring, Birgitte Kiær

1994-01-01

Thermophilic anaerobic processes are often regarded as less stable than mesophilic processes. In the paper this postulate is examined and disproved based on real operational data from of full-scale mesophilic and thermophilic biogas plants. The start-up produce for the thermophilic plants was...... for thermophilic digestion along with the implications for the methanogenic bacteria active at these temperatures....

ANAEROBIC BIODEGRADATION OF A BIODEGRADABLE MATERIAL UNDER ANAEROBIC - THERMOPHILIC DIGESTION

Directory of Open Access Journals (Sweden)

RICARDO CAMACHO-MUÑOZ

2014-12-01

Full Text Available This paper dertermined the anaerobic biodegradation of a polymer obtained by extrusion process of native cassava starch, polylactic acid and polycaprolactone. Initially a thermophilic - methanogenic inoculum was prepared from urban solid waste. The gas final methane concentration and medium’s pH reached values of 59,6% and 7,89 respectively. The assay assembly was carried out according ASTM D5511 standard. The biodegradation percent of used materials after 15 day of digestion were: 77,49%, 61,27%, 0,31% for cellulose, sample and polyethylene respectively. Due cellulose showed biodegradation levels higher than 70% it’s deduced that the inoculum conditions were appropriate. A biodegradation level of 61,27%, 59,35% of methane concentration in sample’s evolved gas and a medium’s finale pH of 7,71 in sample’s vessels, reveal the extruded polymer´s capacity to be anaerobically degraded under thermophilic- high solid concentration conditions.

Modified anaerobic digestion elutriated phased treatment for the anaerobic co-digestion of sewage sludge and food wastewater.

Science.gov (United States)

Mo, Kyung; Lee, Wonbae; Kim, Moonil

2017-02-01

A modified anaerobic digestion elutriated phased treatment (MADEPT) process was developed for investigating anaerobic co-digestion of sewage sludge and food wastewater. The anaerobic digestion elutriated phased treatment (ADEPT) process is similar to a two-phase system, however, in which the effluent from a methanogenic reactor recycles into an acidogenic reactor to elutriate mainly dissolved organics. Although ADEPT could reduce reactor volume significantly, the unsolubilized solids should be wasted from the system. The MADEPT process combines thermo-alkali solubilization with ADEPT to improve anaerobic performance and to minimize the sludge disposal. It was determined that the optimal volume mixing ratio of sewage sludge and food wastewater was 4 : 1 for the anaerobic co-digestion. The removal efficiencies of total chemical oxygen demand, volatile solids, and volatile suspended solids in the MADEPT process were 73%, 70%, and 64%, respectively. However, those in the ADEPT process were only 48%, 37%, and 40%, respectively, at the same hydraulic retention time (HRT) of 7 days. The gas production of MADEPT was two times higher than that of ADEPT. The thermo-alkali solubilization increased the concentration of dissolved organics so that they could be effectively degraded in a short HRT, implying that MADEPT could improve the performance of ADEPT in anaerobic co-digestion.

Anaerobic electrochemical membrane bioreactor and process for wastewater treatment

KAUST Repository

Amy, Gary

2015-07-09

An anaerobic electrochemical membrane bioreactor (AnEMBR) can include a vessel into which wastewater can be introduced, an anode electrode in the vessel suitable for supporting electrochemically active microorganisms (EAB, also can be referred to as anode reducing bacteria, exoelectrogens, or electricigens) that oxidize organic compounds in the wastewater, and a cathode membrane electrode in the vessel, which is configured to pass a treated liquid through the membrane while retaining the electrochemically active microorganisms and the hydrogenotrophic methanogens (for example, the key functional microbial communities, including EAB, methanogens and possible synergistic fermenters) in the vessel. The cathode membrane electrode can be suitable for catalyzing the hydrogen evolution reaction to generate hydro en.

Isolation and Partial Characterization of Bacteria in an Anaerobic Consortium That Mineralizes 3-Chlorobenzoic Acid †

OpenAIRE

Shelton, Daniel R.; Tiedje, James M.

1984-01-01

A methanogenic consortium able to use 3-chlorobenzoic acid as its sole energy and carbon source was enriched from anaerobic sewage sludge. Seven bacteria were isolated from the consortium in mono- or coculture. They included: one dechlorinating bacterium (strain DCB-1), one benzoate-oxidizing bacterium (strain BZ-2), two butyrate-oxidizing bacteria (strains SF-1 and NSF-2), two H2-consuming methanogens (Methanospirillum hungatei PM-1 and Methanobacterium sp. strain PM-2), and a sulfate-reduci...

Comparison of different liquid anaerobic digestion effluents as inocula and nitrogen sources for solid-state batch anaerobic digestion of corn stover

International Nuclear Information System (INIS)

Xu Fuqing; Shi Jian; Lv Wen; Yu Zhongtang; Li Yebo

2013-01-01

Highlights: ► Compared methane production of solid AD inoculated with different effluents. ► Food waste effluent (FWE) had the largest population of acetoclastic methanogens. ► Solid AD inoculated with FWE produced the highest methane yield at F/E ratio of 4. ► Dairy waste effluent (DWE) was rich of cellulolytic and xylanolytic bacteria. ► Solid AD inoculated with DWE produced the highest methane yield at F/E ratio of 2. - Abstract: Effluents from three liquid anaerobic digesters, fed with municipal sewage sludge, food waste, or dairy waste, were evaluated as inocula and nitrogen sources for solid-state batch anaerobic digestion of corn stover in mesophilic reactors. Three feedstock-to-effluent (F/E) ratios (i.e., 2, 4, and 6) were tested for each effluent. At an F/E ratio of 2, the reactor inoculated by dairy waste effluent achieved the highest methane yield of 238.5 L/kgVS feed , while at an F/E ratio of 4, the reactor inoculated by food waste effluent achieved the highest methane yield of 199.6 L/kgVS feed . The microbial population and chemical composition of the three effluents were substantially different. Food waste effluent had the largest population of acetoclastic methanogens, while dairy waste effluent had the largest populations of cellulolytic and xylanolytic bacteria. Dairy waste also had the highest C/N ratio of 8.5 and the highest alkalinity of 19.3 g CaCO 3 /kg. The performance of solid-state batch anaerobic digestion reactors was closely related to the microbial status in the liquid anaerobic digestion effluents.

Comparison of different liquid anaerobic digestion effluents as inocula and nitrogen sources for solid-state batch anaerobic digestion of corn stover

Energy Technology Data Exchange (ETDEWEB)

Xu Fuqing; Shi Jian [Department of Food, Agricultural and Biological Engineering, Ohio State University, Ohio Agricultural Research and Development Center, 1680 Madison Ave., Wooster, OH 44691 (United States); Lv Wen; Yu Zhongtang [Department of Animal Sciences, Ohio State University, Columbus, OH 43210 (United States); Li Yebo, E-mail: li.851@osu.edu [Department of Food, Agricultural and Biological Engineering, Ohio State University, Ohio Agricultural Research and Development Center, 1680 Madison Ave., Wooster, OH 44691 (United States)

2013-01-15

Highlights: Black-Right-Pointing-Pointer Compared methane production of solid AD inoculated with different effluents. Black-Right-Pointing-Pointer Food waste effluent (FWE) had the largest population of acetoclastic methanogens. Black-Right-Pointing-Pointer Solid AD inoculated with FWE produced the highest methane yield at F/E ratio of 4. Black-Right-Pointing-Pointer Dairy waste effluent (DWE) was rich of cellulolytic and xylanolytic bacteria. Black-Right-Pointing-Pointer Solid AD inoculated with DWE produced the highest methane yield at F/E ratio of 2. - Abstract: Effluents from three liquid anaerobic digesters, fed with municipal sewage sludge, food waste, or dairy waste, were evaluated as inocula and nitrogen sources for solid-state batch anaerobic digestion of corn stover in mesophilic reactors. Three feedstock-to-effluent (F/E) ratios (i.e., 2, 4, and 6) were tested for each effluent. At an F/E ratio of 2, the reactor inoculated by dairy waste effluent achieved the highest methane yield of 238.5 L/kgVS{sub feed}, while at an F/E ratio of 4, the reactor inoculated by food waste effluent achieved the highest methane yield of 199.6 L/kgVS{sub feed}. The microbial population and chemical composition of the three effluents were substantially different. Food waste effluent had the largest population of acetoclastic methanogens, while dairy waste effluent had the largest populations of cellulolytic and xylanolytic bacteria. Dairy waste also had the highest C/N ratio of 8.5 and the highest alkalinity of 19.3 g CaCO{sub 3}/kg. The performance of solid-state batch anaerobic digestion reactors was closely related to the microbial status in the liquid anaerobic digestion effluents.

Simple and versatile turbidimetric monitoring of bacterial growth in liquid cultures using a customized 3D printed culture tube holder and a miniaturized spectrophotometer: application to facultative and strictly anaerobic bacteria

Directory of Open Access Journals (Sweden)

Margarida R. G. Maia

2016-08-01

Full Text Available Here we introduce a novel strategy for turbidimetric monitoring of bacterial growth in liquid culture. The instrumentation comprises a light source, a customized 3D printed culture tube holder and a miniaturized spectrophotometer, connected through optical cables. Due to its small footprint and the possibility to operate with external light, bacterial growth was directly monitored from culture tubes in a simple and versatile fashion. This new portable measurement technique was used to monitor the growth of facultative (Escherichia coli ATCC/25922, and Staphylococcus aureus ATCC/29213 and strictly (Butyrivibrio fibrisolvens JW11, Butyrivibrio proteoclasticus P18, and Propionibacterium acnes DSMZ 1897 anaerobic bacteria. For E. coli and S. aureus, the growth rates calculated from normalized optical density values were compared with those ones obtained using a benchtop spectrophotometer without significant differences (P = 0.256. For the strictly anaerobic species, a high precision (RSD < 3.5% was observed between replicates up to 48 h. Regarding its potential for customization, this manifold could accommodate further developments for customized turbidimetric monitoring, such as the use of light-emitting diodes as a light source or flow cells.

Simple and Versatile Turbidimetric Monitoring of Bacterial Growth in Liquid Cultures Using a Customized 3D Printed Culture Tube Holder and a Miniaturized Spectrophotometer: Application to Facultative and Strictly Anaerobic Bacteria.

Science.gov (United States)

Maia, Margarida R G; Marques, Sara; Cabrita, Ana R J; Wallace, R John; Thompson, Gertrude; Fonseca, António J M; Oliveira, Hugo M

2016-01-01

Here we introduce a novel strategy for turbidimetric monitoring of bacterial growth in liquid culture. The instrumentation comprises a light source, a customized 3D printed culture tube holder and a miniaturized spectrophotometer, connected through optical cables. Due to its small footprint and the possibility to operate with external light, bacterial growth was directly monitored from culture tubes in a simple and versatile fashion. This new portable measurement technique was used to monitor the growth of facultative (Escherichia coli ATCC/25922, and Staphylococcus aureus ATCC/29213) and strictly (Butyrivibrio fibrisolvens JW11, Butyrivibrio proteoclasticus P18, and Propionibacterium acnes DSMZ 1897) anaerobic bacteria. For E. coli and S. aureus, the growth rates calculated from normalized optical density values were compared with those ones obtained using a benchtop spectrophotometer without significant differences (P = 0.256). For the strictly anaerobic species, a high precision (relative standard deviation < 3.5%) was observed between replicates up to 48 h. Regarding its potential for customization, this manifold could accommodate further developments for customized turbidimetric monitoring, such as the use of light-emitting diodes as a light source or flow cells.

Identification of novel potential acetate-oxidizing bacteria in an acetate-fed methanogenic chemostat based on DNA stable isotope probing.

Science.gov (United States)

Wang, Hui-Zhong; Gou, Min; Yi, Yue; Xia, Zi-Yuan; Tang, Yue-Qin

2018-05-11

Acetate is a significant intermediate of anaerobic fermentation. There are two pathways for converting acetate to CH 4 and CO 2 : acetoclastic methanogenesis by acetoclastic methanogens, and syntrophic acetate oxidation by acetate-oxidizing bacteria (AOB) and hydrogenotrophic methanogens. Detailed investigations of syntrophic acetate-oxidizing bacteria (SAOB) should contribute to the elucidation of the microbial mechanisms of methanogenesis. In this study, we investigated the major phylogenetic groups of acetate-utilizing bacteria (AUB) in a mesophilic methanogenic chemostat fed with acetate as the sole carbon source by using DNA stable isotope probing (SIP) technology. The results indicated that acetoclastic methanogenesis and acetate oxidization/hydrogenotrophic methanogenesis coexisted in the mesophilic chemostat fed with acetate, operated at a dilution rate of 0.1 d -1 . OTU Ace13(9-17) (KU869530), Ace13(9-4) (KU667241), and Ace13(9-23) (KU667236), assigned to the phyla Firmicutes and Bacteroidetes, were probably potential SAOB in the chemostat, which needs further investigation. Species in the phyla Proteobacteria, Deferribacteres, Acidobacteria, Spirochaetes and Actinobacteria were probably capable of utilizing acetate for their growth. Methanoculleus was likely to be the preferred hydrogenotrophic methanogen for syntrophy with AOB in the chemostat.

Bacteria and archaea communities in full-scale thermophilic and mesophilic anaerobic digesters treating food wastewater: Key process parameters and microbial indicators of process instability.

Science.gov (United States)

Lee, Joonyeob; Shin, Seung Gu; Han, Gyuseong; Koo, Taewoan; Hwang, Seokhwan

2017-12-01

In this study, four different mesophilic and thermophilic full-scale anaerobic digesters treating food wastewater (FWW) were monitored for 1-2years in order to investigate: 1) microbial communities underpinning anaerobic digestion of FWW, 2) significant factors shaping microbial community structures, and 3) potential microbial indicators of process instability. Twenty-seven bacterial genera were identified as abundant bacteria underpinning the anaerobic digestion of FWW. Methanosaeta harundinacea, M. concilii, Methanoculleus bourgensis, M. thermophilus, and Methanobacterium beijingense were revealed as dominant methanogens. Bacterial community structures were clearly differentiated by digesters; archaeal community structures of each digester were dominated by one or two methanogen species. Temperature, ammonia, propionate, Na + , and acetate in the digester were significant factors shaping microbial community structures. The total microbial populations, microbial diversity, and specific bacteria genera showed potential as indicators of process instability in the anaerobic digestion of FWW. Copyright © 2017 Elsevier Ltd. All rights reserved.

Bacterial ecology of abattoir wastewater treated by an anaerobic digestor.

Science.gov (United States)

Jabari, Linda; Gannoun, Hana; Khelifi, Eltaief; Cayol, Jean-Luc; Godon, Jean-Jacques; Hamdi, Moktar; Fardeau, Marie-Laure

2016-01-01

Wastewater from an anaerobic treatment plant at a slaughterhouse was analysed to determine the bacterial biodiversity present. Molecular analysis of the anaerobic sludge obtained from the treatment plant showed significant diversity, as 27 different phyla were identified. Firmicutes, Proteobacteria, Bacteroidetes, Thermotogae, Euryarchaeota (methanogens), and msbl6 (candidate division) were the dominant phyla of the anaerobic treatment plant and represented 21.7%, 18.5%, 11.5%, 9.4%, 8.9%, and 8.8% of the total bacteria identified, respectively. The dominant bacteria isolated were Clostridium, Bacteroides, Desulfobulbus, Desulfomicrobium, Desulfovibrio and Desulfotomaculum. Our results revealed the presence of new species, genera and families of microorganisms. The most interesting strains were characterised. Three new bacteria involved in anaerobic digestion of abattoir wastewater were published. Copyright © 2015 Sociedade Brasileira de Microbiologia. Published by Elsevier Editora Ltda. All rights reserved.

Effects of lipid concentration on anaerobic co-digestion of municipal biomass wastes

International Nuclear Information System (INIS)

Sun, Yifei; Wang, Dian; Yan, Jiao; Qiao, Wei; Wang, Wei; Zhu, Tianle

2014-01-01

Highlights: • Lipid in municipal biomass would not inhibited the anaerobic digestion process. • A lipid concentration of 65% of total VS was the inhibition concentration. • The amount of Brevibacterium decreased with the increasing of the lipid contents. • Long chain fatty acids stacked on the methanogenic bacteria and blocked the mass transfer process. - Abstract: The influence of the lipid concentration on the anaerobic co-digestion of municipal biomass waste and waste-activated sludge was assessed by biochemical methane potential (BMP) tests and by bench-scale tests in a mesophilic semi-continuous stirred tank reactor. The effect of increasing the volatile solid (VS) concentration of lipid from 0% to 75% was investigated. BMP tests showed that lipids in municipal biomass waste could enhance the methane production. The results of bench-scale tests showed that a lipids concentration of 65% of total VS was the inhibition concentration. Methane yields increased with increasing lipid concentration when lipid concentrations were below 60%, but when lipid concentration was set as 65% or higher, methane yields decreased sharply. When lipid concentrations were below 60%, the pH values were in the optimum range for the growth of methanogenic bacteria and the ratios of volatile fatty acid (VFA)/alkalinity were in the range of 0.2–0.6. When lipid concentrations exceeded 65%, the pH values were below 5.2, the reactor was acidized and the values of VFA/alkalinity rose to 2.0. The amount of Brevibacterium decreased with increasing lipid content. Long chain fatty acids stacked on the methanogenic bacteria and blocked the mass transfer process, thereby inhibiting anaerobic digestion

Effects of lipid concentration on anaerobic co-digestion of municipal biomass wastes

Energy Technology Data Exchange (ETDEWEB)

Sun, Yifei, E-mail: sunif@buaa.edu.cn [School of Chemistry and Environment, Beihang University, Beijing 100191 (China); Wang, Dian; Yan, Jiao [School of Chemistry and Environment, Beihang University, Beijing 100191 (China); Qiao, Wei [College of Chemical Science and Engineering, China University of Petroleum, Beijing 102249 (China); Wang, Wei [School of Environment, Tsinghua University, Beijing 100084 (China); Zhu, Tianle [School of Chemistry and Environment, Beihang University, Beijing 100191 (China)

2014-06-01

Highlights: • Lipid in municipal biomass would not inhibited the anaerobic digestion process. • A lipid concentration of 65% of total VS was the inhibition concentration. • The amount of Brevibacterium decreased with the increasing of the lipid contents. • Long chain fatty acids stacked on the methanogenic bacteria and blocked the mass transfer process. - Abstract: The influence of the lipid concentration on the anaerobic co-digestion of municipal biomass waste and waste-activated sludge was assessed by biochemical methane potential (BMP) tests and by bench-scale tests in a mesophilic semi-continuous stirred tank reactor. The effect of increasing the volatile solid (VS) concentration of lipid from 0% to 75% was investigated. BMP tests showed that lipids in municipal biomass waste could enhance the methane production. The results of bench-scale tests showed that a lipids concentration of 65% of total VS was the inhibition concentration. Methane yields increased with increasing lipid concentration when lipid concentrations were below 60%, but when lipid concentration was set as 65% or higher, methane yields decreased sharply. When lipid concentrations were below 60%, the pH values were in the optimum range for the growth of methanogenic bacteria and the ratios of volatile fatty acid (VFA)/alkalinity were in the range of 0.2–0.6. When lipid concentrations exceeded 65%, the pH values were below 5.2, the reactor was acidized and the values of VFA/alkalinity rose to 2.0. The amount of Brevibacterium decreased with increasing lipid content. Long chain fatty acids stacked on the methanogenic bacteria and blocked the mass transfer process, thereby inhibiting anaerobic digestion.

Methanogenic Paraffin Biodegradation: Alkylsuccinate Synthase Gene Quantification and Dicarboxylic Acid Production.

Science.gov (United States)

Oberding, Lisa K; Gieg, Lisa M

2018-01-01

applications for effective fuel-contaminated site remediation and for improved recovery from oil reservoirs. Previous studies have clearly demonstrated that short-chain alkanes (C 17 ) that comprise many fuel mixtures. Using an enrichment culture derived from a freshwater fuel-contaminated site, we demonstrate that the model waxy alkane n -octacosane can be biodegraded under methanogenic conditions by a presumed Smithella phylotype. Compared with that of controls, we show an increased abundance and expression of the assA gene, which is known to be important for anaerobic n -alkane metabolism. Metabolite analyses revealed the presence of a range of α,ω-dicarboxylic acids found only in n -octacosane-degrading cultures, a novel finding that lends insight as to how anaerobic communities may access waxes as growth substrates in anoxic environments. Copyright © 2017 American Society for Microbiology.

Methanogenic H2 syntrophy among thermophiles: a model of metabolism, adaptation and survival in the subsurface

Science.gov (United States)

Topcuoglu, B. D.; Stewart, L. C.; Butterfield, D. A.; Huber, J. A.; Holden, J. F.

2016-12-01

Approximately 1 giga ton (Gt, 1015 g) of CH4 is formed globally per year from H2, CO2 and acetate through methanogenesis, largely by methanogens growing in syntrophic association with anaerobic microbes that hydrolyze and ferment biopolymers. However, our understanding of methanogenesis in hydrothermal regions of the subseafloor and potential syntrophic methanogenesis at thermophilic temperatures (i.e., >50°C) is nascent. In this study, the growth of natural assemblages of thermophilic methanogens from Axial Seamount was primarily limited by H2 availability. Heterotrophs supported thermophilic methanogenesis by H2 syntrophy in microcosm incubations of hydrothermal fluids at 55°C and 80°C supplemented with tryptone only. Based on 16S rRNA gene sequencing, only heterotrophic archaea that produce H2, H2-consuming methanogens, and sulfate reducing archaea were found in 80°C tryptone microcosms from Marker 113 vent. No bacteria were found. In 55°C tryptone microcosms, sequences were found from H2-producing bacteria and H2-consuming methanogens and sulfate-reducing bacteria. In order to model the impact of H2 syntrophy at hyperthemophilic temperatures, a co-culture was established consisting of the H2-producing hyperthermophilic heterotroph Thermococcus paralvinellae and a H2-consuming hyperthermophilic methanogen Methanocaldococcus bathoardescens. When grown alone in a chemostat, the growth rates and steady-state cell concentrations of T. paralvinellae decreased significantly when a high H2 (70 µM) background was present. H2 inhibition was ameliorated by the production of formate, but in silico modeling suggests less energetic yield for the cells. H2 syntrophy relieved H2 inhibition for both the heterotroph and the methanogenic partners. The results demonstrate that thermophilic H2 syntrophy can support methanogenesis within natural microbial assemblages and may be an important alternative energy source for thermophilic autotrophs in marine geothermal environments.

[Effects of selective methanogenic inhibitors on methanogenesis and methanogenic communities in acetate degrading cultures].

Science.gov (United States)

Ma, Tingting; Cheng, Lei; Liu, Laiyan; Dai, Lirong; Zhou, Zheng; Zhang, Hui

2015-05-04

We evaluated the role of syntrophic acetate oxidation coupled with hydrogenotrophic methanogens in three different methanogenic consortia. Three methanogenic hexadecane degrading consortia named Y15, M82 and SK were taken from the same oily sludge of Shengli oil-field and enriched. They were incubated at 15, 35 and 55 °C, respectively. The consortia amended with acetate and inhibitors of NH4Cl or CH3F were further transferred and incubated at corresponding temperatures. The cultures atlate logarithmic phase were collected for terminal restriction fragment length polymorphism (T-RFLP) combined with cloning and phylogenetic analysis of 16S rRNA gene fragments. Gas chromatograph analysis showed that all of the consortia could grow and produce methane, but the lag phase was delayed and the growth rate was retarded in the cultures amended with inhibitor. Combination analysis of T-RFLP and clone library revealed the predominance of obligate aceticlastic Methanosaeta in the acetate cultures of Y15, M82 and SK. Under the mesophilic and thermophilic conditions, after add inginhibitor the relative abundance of aceticlastic methanogen decreased but hydrogenotrophic methanogen increased. Syntrophic acetate oxidation during methanogenic degradation of petroleum hydrocarbons occurs under mesophilic and thermophilic conditions, although the situation at low temperature seems uncertain.

The fate of methanol in anaerobic bioreactors

OpenAIRE

Florencio, L.

1994-01-01

Methanol is an important component of certain industrial wastewaters. In anaerobic environments, methanol can be utilized by methanogens and acetogens. In wastewater treatment plants, the conversion of methanol into methane is preferred because this conversion is responsible for chemical oxygen demand (COD) removal, whereas with the formation of volatile fatty acids (VFA) little COD removal is achieved. Moreover, the accumulation of VFA can lead to reactor instability due to pH drops...

Assessment of the anaerobic degradation of six active pharmaceutical ingredients.

Science.gov (United States)

Musson, Stephen E; Campo, Pablo; Tolaymat, Thabet; Suidan, Makram; Townsend, Timothy G

2010-04-01

Research examined the anaerobic degradation of 17 alpha-ethynylestradiol, acetaminophen, acetylsalicylic acid, ibuprofen, metoprolol tartrate, and progesterone by methanogenic bacteria. Using direct sample analysis and respirometric testing, anaerobic degradation was examined with (a) each compound as the sole organic carbon source and (b) each compound at a lower concentration (250 microg/L) and cellulose serving as the primary organic carbon source. The change in pharmaceutical concentration was determined following 7, 28, 56, and 112 days of anaerobic incubation at 37 degrees C. Only acetylsalicylic acid demonstrated significant degradation; the remaining compounds showed a mixture of degradation and abiotic removal mechanisms. Experimental results were compared with BIOWIN, an anaerobic degradation prediction model of the US Environmental Protection Agency. The BIOWIN model predicted anaerobic biodegradability of the compounds in the order: acetylsalicylic acid > metoprolol tartrate > ibuprofen > acetaminophen > 17 alpha-ethinylestradiol >progesterone. This corresponded well with the experimental findings which found degradability in the order: acetylsalicylic acid > metoprolol tartrate > acetaminophen > ibuprofen. (c) 2010 Elsevier B.V. All rights reserved.

Methanization potential of anaerobic biodigestion of solid food waste

Directory of Open Access Journals (Sweden)

Laís R. G. de Oliveira

Full Text Available ABSTRACT Anaerobic biodigestion of solid and semi-solid wastes has been widely used for the treatment of these residues and methane production; however, during the process (more specifically in the acidogenic phase, there is a tendency of pH reduction, an unfavorable condition to methanogenic bacteria. Thus, the present work aims to evaluate the methanization potential of an agroindustrial anaerobic granular sludge (AIS from UASB (Upflow Anaerobic Sludge Blanket reactor, individually and biodigested with food waste (FW from the University Restaurant of the Federal University of Pernambuco with buffering agent (AIS + FW + b and without it (AIS + FW. After the laboratory tests, the AIS + FW + b configuration obtained a cumulative methane production approximately six times greater than that of AIS + FW, and approximately twice that of the inoculum alone (AIS.

Quantitation and identification of methanogens and sulphate reducers in Olkiluoto groundwater

International Nuclear Information System (INIS)

Bomberg, M.; Nyyssoenen, M.; Itaevaara, M.

2010-08-01

The GEOFUNC Project focuses on the microbiology connected to safety and risk assessment of the final disposal of high radioactive nuclear waste. Methanogenic archaea and sulphate reducing bacteria are significant groups of microorganisms in anaerobic environments, and are of crucial concern for the safe long term storage of nuclear waste in deep bedrock. The sulphate reducing bacteria are able to produce sulphide which may cause corrosion of the copper in the radioactive waste storage capsules. Methanogens, on the other hand, may produce quantities of methane from various organic carbon compounds, CO 2 and H 2 . Methane may both serve as carbon source for methanotrophic microbial groups, and may also cause mobilization of radionuclides, as a result of gas discharge through fractures in the bedrock. The transition zones between the sulphate rich and methane rich waters are locations for microbial processes where the methane may serve as carbon source for sulphate reducing bacteria, which in turn would produce corrosive sulphides. It has been estimated that only 1-10 % of all the microorganisms present in the environment can be isolated and cultivated. Uncultured microorganisms can be identified and their numbers in the environment quantified by identification of specific marker genes that are essential for their functions by use of molecular methods. Methanogens, for example, can be identified by their genes for methyl coenzyme M reductase (mcrA), which is an essential enzyme involved in the production of methane. The mcrA is specifically present only in methanogenic archaea. Sulphate reducers are identified by their dissimilatory sulphite reductase genes (dsrB), which are present in and essential for all microorganisms performing dissimilatory sulphate reducing. In the GEOFUNC project, a quantitative PCR method (qPCR) was developed for the detection of methanogens and sulphate reducers. This method is based on specific quantitative detection of marker genes

Effects of oxytetracycline, tylosin, and amoxicillin antibiotics on specific methanogenic activity of anaerobic biomass

OpenAIRE

Mohammad Mehdi Amin; Hassan Hashemi; Afshin Ebrahimi; Asghar Ebrahimi

2012-01-01

Aims: The purpose of this study was to survey the antibiotics effects of oxytetracycline, tylosin, and amoxicillin on anerobic wastewater treatment process. Materials and Methods: To evaluate the inhibitory antibiotics amoxicillin, tetracycline, and tylosin on biomass activity, specific methanogenic activity (SMA) using anerobic biomass batch; into 120 ml vials: 30 ml biomass and 70 ml substrate including volatile fatty acids, mainly acetic acid and various concentrations of antibiotics we...

Effects of microbial inhibitors on anaerobic degradation of DDT

Energy Technology Data Exchange (ETDEWEB)

Wang, Y.S.; Chiu, T.C.; Yen, J.H. [National Taiwan Univ., Taipei (Taiwan)

2004-09-15

Chlorinated insecticide DDT [1,1,1-trichloro-2,2-bis(p-chlorophenyl)ethane] was extensively used for controlling pests in the agricultural field and human-being living environments in the past several decades. Due to the chemical stability, DDT was extremely persistent and recalcitrant in soils and sediments and it was banned by nations. Microorganisms usually play important roles in reducing organochlorine compounds in the environments. Under low-oxygen conditions, microbial dechlorination is thought as the onset of highly chlorinated compounds. Methanogenic and sulfate-reducing bacteria participate in microbial dechlorination under anaerobic condition has been reported. In this study, a mixed anaerobic culture enabling to dechlorinate DDT was obtained from river sediment in Taiwan. In order to understand the effect of these microorganisms on DDT dechlorination, microbial inhibitors BESA (2-bromoethanesulfonate) and molybdate, for inhibiting methanogenic and sulfate-reducing bacteria, respectively, were chosen to investigate the interaction between specific microbial communities and their degradation activities. Besides, a molecular technique, denaturing gradient gel electrophoresis (DGGE), based on analyzing the 16S rDNA of bacteria, was used for monitoring the bacterial community structure in this study.

Biomethanation Of Syngas Using Anaerobic Sludge: Shift In The Catabolic Routes With The CO Partial Pressure Increase

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Silvia Sancho-Navarro

2016-08-01

Full Text Available Syngas generated by thermal gasification of biomass or coal can be steam reformed and purified into methane, which could be used locally for energy needs, or re-injected in the natural gas grid. As an alternative to chemical catalysis, the main components of the syngas (CO, CO2, and H2 can be used as substrates by a wide range of microorganisms, to be converted into gas biofuels, including methane. This study evaluates the carboxydotrophic (CO-consuming methanogenic potential present in an anaerobic sludge from an upflow anaerobic sludge bed (UASB reactor treating waste water, and elucidates the CO conversion routes to methane at 35±3˚C. Kinetic activity tests under CO at partial pressures (pCO varying from 0.1 to 1.5 atm (0.09-1.31 mmol/L in the liquid phase showed a significant carboxydotrophic activity potential for growing conditions on CO alone. A maximum methanogenic activity of 1 mmol CH4 per g of volatile suspended solid and per day was achieved at 0.2 atm of CO (0.17 mmol/L, and then the rate decreased with the amount of CO supplied. The intermediary metabolites such as acetate, H2 and propionate started to accumulate at higher CO concentrations. Inhibition experiments with 2-bromoethanesulfonic acid (BES, fluoroacetate, and vancomycin showed that in a mixed culture CO was converted mainly to acetate by acetogenic bacteria, which was further transformed to methane by acetoclastic methanogens, while direct methanogenic CO conversion was negligible. Methanogenesis was totally blocked at high pCO in the bottles (≥ 1 atm. However it was possible to achieve higher methanogenic potential under a 100% CO atmosphere after acclimation of the sludge to CO. This adaptation to high CO concentrations led to a shift in the archaeal population, then dominated by hydrogen-utilizing methanogens, which were able to take over acetoclastic methanogens, while syntrophic acetate oxidizing (SAO bacteria oxidized acetate into CO2 and H2. The disaggregation

Biomethanation of Syngas Using Anaerobic Sludge: Shift in the Catabolic Routes with the CO Partial Pressure Increase.

Science.gov (United States)

Sancho Navarro, Silvia; Cimpoia, Ruxandra; Bruant, Guillaume; Guiot, Serge R

2016-01-01

Syngas generated by thermal gasification of biomass or coal can be steam reformed and purified into methane, which could be used locally for energy needs, or re-injected in the natural gas grid. As an alternative to chemical catalysis, the main components of the syngas (CO, CO2, and H2) can be used as substrates by a wide range of microorganisms, to be converted into gas biofuels, including methane. This study evaluates the carboxydotrophic (CO-consuming) methanogenic potential present in an anaerobic sludge from an upflow anaerobic sludge bed (UASB) reactor treating waste water, and elucidates the CO conversion routes to methane at 35 ± 3°C. Kinetic activity tests under CO at partial pressures (pCO) varying from 0.1 to 1.5 atm (0.09-1.31 mmol/L in the liquid phase) showed a significant carboxydotrophic activity potential for growing conditions on CO alone. A maximum methanogenic activity of 1 mmol CH4 per g of volatile suspended solid and per day was achieved at 0.2 atm of CO (0.17 mmol/L), and then the rate decreased with the amount of CO supplied. The intermediary metabolites such as acetate, H2, and propionate started to accumulate at higher CO concentrations. Inhibition experiments with 2-bromoethanesulfonic acid (BES), fluoroacetate, and vancomycin showed that in a mixed culture CO was converted mainly to acetate by acetogenic bacteria, which was further transformed to methane by acetoclastic methanogens, while direct methanogenic CO conversion was negligible. Methanogenesis was totally blocked at high pCO in the bottles (≥1 atm). However it was possible to achieve higher methanogenic potential under a 100% CO atmosphere after acclimation of the sludge to CO. This adaptation to high CO concentrations led to a shift in the archaeal population, then dominated by hydrogen-utilizing methanogens, which were able to take over acetoclastic methanogens, while syntrophic acetate oxidizing (SAO) bacteria oxidized acetate into CO2 and H2. The disaggregation of the

Growth of desulfovibrio in lactate or ethanol media low in sulfate in association with H2-utilizing methanogenic bacteria.

Science.gov (United States)

Bryant, M P; Campbell, L L; Reddy, C A; Crabill, M R

1977-05-01

In the analysis of an ethanol-CO(2) enrichment of bacteria from an anaerobic sewage digestor, a strain tentatively identified as Desulfovibrio vulgaris and an H(2)-utilizing methanogen resembling Methanobacterium formicicum were isolated, and they were shown to represent a synergistic association of two bacterial species similar to that previously found between S organism and Methanobacterium strain MOH isolated from Methanobacillus omelianskii. In lowsulfate media, the desulfovibrio produced acetate and H(2) from ethanol and acetate, H(2), and, presumably, CO(2) from lactate; but growth was slight and little of the energy source was catabolized unless the organism was combined with an H(2)-utilizing methanogenic bacterium. The type strains of D. vulgaris and Desulfovibrio desulfuricans carried out the same type of synergistic growth with methanogens. In mixtures of desulfovibrio and strain MOH growing on ethanol, lactate, or pyruvate, diminution of methane produced was stoichiometric with the moles of sulfate added, and the desulfovibrios grew better with sulfate addition. The energetics of the synergistic associations and of the competition between the methanogenic system and sulfate-reducing system as sinks for electrons generated in the oxidation of organic materials such as ethanol, lactate, and acetate are discussed. It is suggested that lack of availability of H(2) for growth of methanogens is a major factor in suppression of methanogenesis by sulfate in natural ecosystems. The results with these known mixtures of bacteria suggest that hydrogenase-forming, sulfate-reducing bacteria could be active in some methanogenic ecosystems that are low in sulfate.

Effect of Cobalt Sorption on Metal Fractionation in Anaerobic Granular Sludge

NARCIS (Netherlands)

Osuna, M.B.; Hullebusch, van E.D.; Zandvoort, M.H.; Iza, J.M.; Lens, P.N.L.

2004-01-01

A sequential extraction procedure was applied to two anaerobic methanogenic sludges (Eerbeek and Nedalco) to examine the speciation of micro- and macronutrients in the sludges after cobalt sorption by exposing the sludge to a 1 mM Co solution for 4 d at pH 7 and 30degreesC. The effect of different

MICROBIAL DEGRADATION OF NITROGEN, OXYGEN AND SULFUR HETEROCYCLIC COMPOUNDS UNDER ANAEROBIC CONDITIONS: STUDIES WITH AQUIFER SAMPLES

Science.gov (United States)

The potential for anaerobic biodegradation of 12 heterocyclic model compounds was studied. Nine of the model compounds were biotransformed in aquifer slurries under sulfate-reducing or methanogenic conditions. The nitrogen and oxygen heterocyclic compounds were more susceptible t...

Efficacies of Various Anaerobic Starter Seeds for Biogas Production from Different Types of Wastewater

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Pawinee Chaiprasert

2017-01-01

Full Text Available Various anaerobic starter seeds from different sources were investigated for their efficacies in treatment of different types of wastewater. Six combinations of starter seeds and wastewaters were selected out of 25 combination batch experiments and operated in semicontinuous reactors. It was noticed that the efficacies of various anaerobic starter seeds for biogas production from different types of wastewater in terms of reactor performance and stability were depended on wastewater characteristics and F/M ratio affecting microbial community and their microbial activities. However, exogenous starter seed can be used across different types of wastewater with or without acclimatization. Four reactors reached the targeted OLR of 2 kg COD/m3·d with high performance and stability except for concentrated rubber wastewater (RBw, even using high active starter seeds of cassava starch (CSs and palm oil (POs. The toxic compounds in RBw such as ammonia and sulfate might also adversely affect methanogenic activity in CSsRBw and POsRBw reactors. DGGE analysis showed that propionate utilizers, Smithella propionica strain LYP and Syntrophus sp., were detected in all samples. For Archaea domain, methylotrophic, hydrogenotrophic, and acetoclastic methanogens were also detected. Syntrophic relationships were assumed between propionate utilizers and methanogens as acetate/H2 producers and utilizers, respectively.

Associative patterns among anaerobic fungi, methanogenic archaea and bacterial communities in response to changes in diet and age in the rumen of dairy cows

Directory of Open Access Journals (Sweden)

Sanjay eKumar

2015-07-01

Full Text Available The rumen microbiome represents a complex microbial genetic web where bacteria, anaerobic rumen fungi (ARF, protozoa and archaea work in harmony contributing to the health and productivity of ruminants. We hypothesized that the rumen microbiome shifts as the dairy cow advances in lactations and these microbial changes may contribute to differences in productivity between primiparous (first lactation and multiparous (≥ second lactation cows. To this end, we investigated shifts in the ruminal ARF and methanogenic communities in both primiparous (n=5 and multiparous (n=5 cows as they transitioned from a high forage to a high grain diet upon initiation of lactation. A total of 20 rumen samples were extracted for genomic DNA, amplified using archaeal and fungal specific primers, sequenced on a 454 platform and analyzed using QIIME. Community comparisons (Bray-Curtis index revealed the effect of diet (P < 0.01 on ARF composition, while archaeal communities differed between primiparous and multiparous cows (P < 0.05. Among ARF, several lineages were unclassified, however, phylum Neocallimastigomycota showed the presence of three known genera. Abundance of Cyllamyces and Caecomyces shifted with diet whereas, Orpinomyces was influenced by both diet and age. Methanobrevibacter constituted the most dominant archaeal genus across all samples. Co-occurrence analysis incorporating taxa from bacteria, ARF and archaea revealed syntrophic interactions both within and between microbial domains in response to change in diet as well as age of dairy cows. Notably, these interactions were numerous and complex in multiparous cows supporting our hypothesis that the rumen microbiome also matures with age to sustain the growing metabolic needs of the host. This study provides a broader picture of the ARF and methanogenic populations in the rumen of dairy cows and their co-occurrence implicates specific relationships between different microbial domains in response to

Influence of two-phase anaerobic digestion on fate of selected antibiotic resistance genes and class I integrons in municipal wastewater sludge

DEFF Research Database (Denmark)

Wu, Ying; Cui, Erping; Zuo, Yiru

2016-01-01

The response of representative antibiotic resistance genes (ARGs) to lab-scale two-phase (acidogenic/methanogenic phase) anaerobic digestion processes under thermophilic and mesophilic conditions was explored. The associated microbial communities and bacterial pathogens were characterized by 16S ...... for ermB and blaTEM. ARGs patterns were correlated with Proteobacteria and Actinobacteria during the two-phase anaerobic digestion....

Anaerobic biodegradability and toxicity of wastewaters from chlorine and total chlorine-free bleaching of eucalyptus kraft pulps.

NARCIS (Netherlands)

Vidal, G.; Soto, M.; Field, J.; Mendez-Pampin, R.; Lema, J.M.

1997-01-01

Chlorine bleaching effluents are problematic for anaerobic wastewater treatment due to their high methanogenic toxicity and low biodegradability. Presently, alternative bleaching processes are being introduced, such as elemental chlorine-free (ECF) and total chlorine-free (TCF) bleaching. The

Anaerobic degradation of linoleic oleic acids

Energy Technology Data Exchange (ETDEWEB)

Lalman, J.A.; Bagley, D.M.

1999-07-01

The anaerobic degradation of linoleic (C18:2) and oleic (C18:1) acids was examined in batch experiments. By-product distribution depended on both the type of long chain fatty acid added and initial substrate concentration. Major by-products were palmitic (C16), myristic (C14) and acetic acids. Trace quantities of palmitoleic (C16:1) and lauric (C12) acids were observed together with larger amounts of palmitic (C16), myristic (C14) and hexanoic (C6) acids in cultures incubated with 100 mg/L linoleic (C18:2) acid. Bio-hydrogenation of C18 fatty acids was not necessary for the {beta}-oxidation mechanism to proceed. Aceticlastic methanogenic inhibition was observed in cultures inoculated with greater than 50 mg/L linoleic (C18:2) acid. In cultures incubated with greater than 50 mg/L oleic (C18:1) acid, aceticlastic methanogenic inhibition was observed for a short time period.

Magnetic resonance microscopy of iron transport in methanogenic granules

Science.gov (United States)

Bartacek, Jan; Vergeldt, Frank J.; Gerkema, Edo; Jenicek, Pavel; Lens, Piet N. L.; Van As, Henk

2009-10-01

Interactions between anaerobic biofilms and heavy metals such as iron, cobalt or nickel are largely unknown. Magnetic resonance imaging (MRI) is a non-invasive method that allows in situ studies of metal transport within biofilm matrixes. The present study investigates quantitatively the penetration of iron (1.75 mM) bound to ethylenediaminetetraacetate (EDTA) into the methanogenic granules (spherical biofilm). A spatial resolution of 109 × 109 × 218 μm 3 and a temporal resolution of 11 min are achieved with 3D Turbo Spin Echo (TSE) measurements. The longitudinal relaxivity, i.e. the slope the dependence of the relaxation rate (1/ T1) on the concentration of paramagnetic metal ions, was used to measure temporal changes in iron concentration in the methanogenic granules. It took up to 300 min for the iron-EDTA complex ([FeEDTA] 2-) to penetrate into the methanogenic granules (3-4 mm in diameter). The diffusion was equally fast in all directions with irregularities such as diffusion-facilitating channels and diffusion-resistant zones. Despite these irregularities, the overall process could be modeled using Fick's equations for diffusion in a sphere, because immobilization of [FeEDTA] 2- in the granular matrix (or the presence of a reactive barrier) was not observed. The effective diffusion coefficient ( D ejf) of [FeEDTA] 2- was found to be 2.8 × 10 -11 m 2 s -1, i.e. approximately 4% of D ejf of [FeEDTA] 2- in water. The Fickian model did not correspond to the processes taking place in the core of the granule (3-5% of the total volume of the granule), where up to 25% over-saturation by iron (compare to the concentration in the bulk solution) occurred.

Synergistic pretreatment of waste activated sludge using CaO_2 in combination with microwave irradiation to enhance methane production during anaerobic digestion

International Nuclear Information System (INIS)

Wang, Jie; Li, Yongmei

2016-01-01

Highlights: • CaO_2/MW pretreatment synergistically enhanced WAS solubilization and CH_4 production. • MW irradiation facilitated more "·OH generation from CaO_2. • The optimal pretreatment condition for methane production was determined. • The growths of both hydrogenotrophic and acetate-utilizing methanogens were promoted. • The dewaterability of WAS was improved considerably by CaO_2/MW treatment. - Abstract: To investigate the effects of combined calcium peroxide (CaO_2) and microwave pretreatment on anaerobic digestion of waste activated sludge, lab-scale experiments were conducted to measure the solubilization, biodegradation, and dewaterability of the waste activated sludge. Additionally, the synergistic effects between CaO_2 and microwave were studied, and the microbial activity and methanogenic archaea community structure were analyzed. Combined pretreatment considerably facilitated the solubilization and subsequent anaerobic digestion of the waste activated sludge. The optimal pretreatment condition was CaO_2 (0.1 g/gVSS)/microwave (480 W, 2 min) for methane production during the subsequent anaerobic digestion process. Under this condition, 80.2% higher CH_4 accumulation yield was achieved after 16 d of anaerobic digestion when compared with the control. The synergistic effects of CaO_2/microwave pretreatment resulted from the different mechanisms of CaO_2 and microwave treatments. Further, microwave irradiation increased "·OH generation from CaO_2 and significantly alleviated the inhibitory effect of CaO_2 on methanogens. The activities of hydrolytic enzymes and acid-forming enzymes in the waste activated sludge were improved after CaO_2 (0.1 g/gVSS)/microwave (480 W, 2 min) pretreatment. Methanogenesis enzyme activity was also higher after CaO_2 treatment (0.1 g/gVSS)/microwave (480 W, 2 min) following a lag period. Illumina MiSeq sequencing analysis indicated that acetate-utilizing methanogen (Methanosaeta sp.) and H_2/CO_2-utilizing

Methanosarcina Play an Important Role in Anaerobic Co-Digestion of the Seaweed Ulva lactuca: Taxonomy and Predicted Metabolism of Functional Microbial Communities.

Directory of Open Access Journals (Sweden)

Jamie A FitzGerald

Full Text Available Macro-algae represent an ideal resource of third generation biofuels, but their use necessitates a refinement of commonly used anaerobic digestion processes. In a previous study, contrasting mixes of dairy slurry and the macro-alga Ulva lactuca were anaerobically digested in mesophilic continuously stirred tank reactors for 40 weeks. Higher proportions of U. lactuca in the feedstock led to inhibited digestion and rapid accumulation of volatile fatty acids, requiring a reduced organic loading rate. In this study, 16S pyrosequencing was employed to characterise the microbial communities of both the weakest (R1 and strongest (R6 performing reactors from the previous work as they developed over a 39 and 27-week period respectively. Comparing the reactor communities revealed clear differences in taxonomy, predicted metabolic orientation and mechanisms of inhibition, while constrained canonical analysis (CCA showed ammonia and biogas yield to be the strongest factors differentiating the two reactor communities. Significant biomarker taxa and predicted metabolic activities were identified for viable and failing anaerobic digestion of U. lactuca. Acetoclastic methanogens were inhibited early in R1 operation, followed by a gradual decline of hydrogenotrophic methanogens. Near-total loss of methanogens led to an accumulation of acetic acid that reduced performance of R1, while a slow decline in biogas yield in R6 could be attributed to inhibition of acetogenic rather than methanogenic activity. The improved performance of R6 is likely to have been as a result of the large Methanosarcina population, which enabled rapid removal of acetic acid, providing favourable conditions for substrate degradation.

Methanosarcina Play an Important Role in Anaerobic Co-Digestion of the Seaweed Ulva lactuca: Taxonomy and Predicted Metabolism of Functional Microbial Communities.

Science.gov (United States)

FitzGerald, Jamie A; Allen, Eoin; Wall, David M; Jackson, Stephen A; Murphy, Jerry D; Dobson, Alan D W

2015-01-01

Macro-algae represent an ideal resource of third generation biofuels, but their use necessitates a refinement of commonly used anaerobic digestion processes. In a previous study, contrasting mixes of dairy slurry and the macro-alga Ulva lactuca were anaerobically digested in mesophilic continuously stirred tank reactors for 40 weeks. Higher proportions of U. lactuca in the feedstock led to inhibited digestion and rapid accumulation of volatile fatty acids, requiring a reduced organic loading rate. In this study, 16S pyrosequencing was employed to characterise the microbial communities of both the weakest (R1) and strongest (R6) performing reactors from the previous work as they developed over a 39 and 27-week period respectively. Comparing the reactor communities revealed clear differences in taxonomy, predicted metabolic orientation and mechanisms of inhibition, while constrained canonical analysis (CCA) showed ammonia and biogas yield to be the strongest factors differentiating the two reactor communities. Significant biomarker taxa and predicted metabolic activities were identified for viable and failing anaerobic digestion of U. lactuca. Acetoclastic methanogens were inhibited early in R1 operation, followed by a gradual decline of hydrogenotrophic methanogens. Near-total loss of methanogens led to an accumulation of acetic acid that reduced performance of R1, while a slow decline in biogas yield in R6 could be attributed to inhibition of acetogenic rather than methanogenic activity. The improved performance of R6 is likely to have been as a result of the large Methanosarcina population, which enabled rapid removal of acetic acid, providing favourable conditions for substrate degradation.

Performance and microbial community variations of anaerobic digesters under increasing tetracycline concentrations

KAUST Repository

Xiong, Yanghui; Harb, Moustapha; Hong, Pei-Ying

2017-01-01

The impact of different concentrations of tetracycline on the performance of anaerobic treatment was evaluated. Results revealed that for all of the tested tetracycline concentrations, no major sustained impact on methane production was observed. Instead, a significant increase in propionic acid was observed in the reactor subjected to the highest concentration of tetracycline (20 mg/L). Microbial community analyses suggest that an alternative methanogenic pathway, specifically that of methanol-utilizing methanogens, may be important for ensuring the stability of methane production in the presence of high tetracycline concentrations. In addition, the accumulation of propionate was due to an increase in volatile fatty acids (VFA)-producing bacteria coupled with a reduction in propionate utilizers. An increase in the abundance of tetracycline resistance genes associated with ribosomal protection proteins was observed after 30 days of exposure to high concentrations of tetracycline, while other targeted resistance genes showed no significant changes. These findings suggest that anaerobic treatment processes can robustly treat wastewater with varying concentrations of antibiotics while also deriving value-added products and minimizing the dissemination of associated antibiotic resistance genes.

Performance and microbial community variations of anaerobic digesters under increasing tetracycline concentrations

KAUST Repository

Xiong, Yanghui

2017-04-01

The impact of different concentrations of tetracycline on the performance of anaerobic treatment was evaluated. Results revealed that for all of the tested tetracycline concentrations, no major sustained impact on methane production was observed. Instead, a significant increase in propionic acid was observed in the reactor subjected to the highest concentration of tetracycline (20 mg/L). Microbial community analyses suggest that an alternative methanogenic pathway, specifically that of methanol-utilizing methanogens, may be important for ensuring the stability of methane production in the presence of high tetracycline concentrations. In addition, the accumulation of propionate was due to an increase in volatile fatty acids (VFA)-producing bacteria coupled with a reduction in propionate utilizers. An increase in the abundance of tetracycline resistance genes associated with ribosomal protection proteins was observed after 30 days of exposure to high concentrations of tetracycline, while other targeted resistance genes showed no significant changes. These findings suggest that anaerobic treatment processes can robustly treat wastewater with varying concentrations of antibiotics while also deriving value-added products and minimizing the dissemination of associated antibiotic resistance genes.

Performance and microbial community variations of anaerobic digesters under increasing tetracycline concentrations.

Science.gov (United States)

Xiong, Yanghui; Harb, Moustapha; Hong, Pei-Ying

2017-07-01

The impact of different concentrations of tetracycline on the performance of anaerobic treatment was evaluated. Results revealed that for all of the tested tetracycline concentrations, no major sustained impact on methane production was observed. Instead, a significant increase in propionic acid was observed in the reactor subjected to the highest concentration of tetracycline (20 mg/L). Microbial community analyses suggest that an alternative methanogenic pathway, specifically that of methanol-utilizing methanogens, may be important for ensuring the stability of methane production in the presence of high tetracycline concentrations. In addition, the accumulation of propionate was due to an increase in volatile fatty acids (VFA)-producing bacteria coupled with a reduction in propionate utilizers. An increase in the abundance of tetracycline resistance genes associated with ribosomal protection proteins was observed after 30 days of exposure to high concentrations of tetracycline, while other targeted resistance genes showed no significant changes. These findings suggest that anaerobic treatment processes can robustly treat wastewater with varying concentrations of antibiotics while also deriving value-added products and minimizing the dissemination of associated antibiotic resistance genes.

Continuous treatment of N-Methyl-p-nitro aniline (MNA) in an Upflow Anaerobic Sludge Blanket (UASB) bioreactor

Science.gov (United States)

Olivares, Christopher I.; Wang, Junqin; Silva Luna, Carlos D.; Field, Jim A.; Abrell, Leif; Sierra-Alvarez, Reyes

2017-01-01

N-methyl-p-nitroaniline (MNA) is an ingredient of insensitive munitions (IM) compounds that serves as a plasticizer and helps reduce unwanted detonations. As its use becomes widespread, MNA waste streams will be generated, necessitating viable treatment options. We studied MNA biodegradation and its inhibition potential to, a representative anaerobic microbial population in wastewater treatment, methanogens. Anaerobic biodegradation and toxicity assays were performed and an up-flow anaerobic sludge blanket reactor (UASB) was operated to test continuous degradation of MNA. MNA was transformed almost stoichiometrically to N-methyl-p-phenylenediamine (MPD). MPD was not mineralized, however, it was readily autoxidized and polymerized extensively upon aeration at pH = 9. In the UASB reactor, MNA was fully degraded up to a loading rate of 297.5 μM MNA d-1). Regarding toxicity, MNA was very inhibitory to acetoclastic methanogens (IC50 = 103 μM) whereas MPD was much less toxic, causing only 13.9% inhibition at the highest concentration tested (1025 μM). The results taken as a whole indicate that anaerobic sludge can transform MNA to MPD continuously, and that the transformation decreases the cytotoxicity of the parent pollutant. MPD can be removed through extensive polymerization. These insights could help define efficient treatment options for waste streams polluted with MNA. PMID:26454121

Illumina sequencing-based analysis of a microbial community enriched under anaerobic methane oxidation condition coupled to denitrification revealed coexistence of aerobic and anaerobic methanotrophs.

Science.gov (United States)

Siniscalchi, Luciene Alves Batista; Leite, Laura Rabelo; Oliveira, Guilherme; Chernicharo, Carlos Augusto Lemos; de Araújo, Juliana Calabria

2017-07-01

Methane is produced in anaerobic environments, such as reactors used to treat wastewaters, and can be consumed by methanotrophs. The composition and structure of a microbial community enriched from anaerobic sewage sludge under methane-oxidation condition coupled to denitrification were investigated. Denaturing gradient gel electrophoresis (DGGE) analysis retrieved sequences of Methylocaldum and Chloroflexi. Deep sequencing analysis revealed a complex community that changed over time and was affected by methane concentration. Methylocaldum (8.2%), Methylosinus (2.3%), Methylomonas (0.02%), Methylacidiphilales (0.45%), Nitrospirales (0.18%), and Methanosarcinales (0.3%) were detected. Despite denitrifying conditions provided, Nitrospirales and Methanosarcinales, known to perform anaerobic methane oxidation coupled to denitrification (DAMO) process, were in very low abundance. Results demonstrated that aerobic and anaerobic methanotrophs coexisted in the reactor together with heterotrophic microorganisms, suggesting that a diverse microbial community was important to sustain methanotrophic activity. The methanogenic sludge was a good inoculum to enrich methanotrophs, and cultivation conditions play a selective role in determining community composition.

Enhanced waste activated sludge digestion using a submerged anaerobic dynamic membrane bioreactor: performance, sludge characteristics and microbial community

Science.gov (United States)

Yu, Hongguang; Wang, Zhiwei; Wu, Zhichao; Zhu, Chaowei

2016-02-01

Anaerobic digestion (AD) plays an important role in waste activated sludge (WAS) treatment; however, conventional AD (CAD) process needs substantial improvements, especially for the treatment of WAS with low solids content and poor anaerobic biodegradability. Herein, we propose a submerged anaerobic dynamic membrane bioreactor (AnDMBR) for simultaneous WAS thickening and digestion without any pretreatment. During the long-term operation, the AnDMBR exhibited an enhanced sludge reduction and improved methane production over CAD process. Moreover, the biogas generated in the AnDMBR contained higher methane content than CAD process. Stable carbon isotopic signatures elucidated the occurrence of combined methanogenic pathways in the AnDMBR process, in which hydrogenotrophic methanogenic pathway made a larger contribution to the total methane production. It was also found that organic matter degradation was enhanced in the AnDMBR, thus providing more favorable substrates for microorganisms. Pyrosequencing revealed that Proteobacteria and Bacteroidetes were abundant in bacterial communities and Methanosarcina and Methanosaeta in archaeal communities, which played an important role in the AnDMBR system. This study shed light on the enhanced digestion of WAS using AnDMBR technology.

Methanobacterium Dominates Biocathodic Archaeal Communities in Methanogenic Microbial Electrolysis Cells

KAUST Repository

Siegert, Michael

2015-07-06

© 2015 American Chemical Society. Methane is the primary end product from cathodic current in microbial electrolysis cells (MECs) in the absence of methanogenic inhibitors, but little is known about the archaeal communities that develop in these systems. MECs containing cathodes made from different materials (carbon brushes, or plain graphite blocks or blocks coated with carbon black and platinum, stainless steel, nickel, ferrihydrite, magnetite, iron sulfide, or molybdenum disulfide) were inoculated with anaerobic digester sludge and acclimated at a set potential of -600 mV (versus a standard hydrogen electrode). The archaeal communities on all cathodes, except those coated with platinum, were predominated by Methanobacterium (median 97% of archaea). Cathodes with platinum contained mainly archaea most similar to Methanobrevibacter. Neither of these methanogens were abundant (<0.1% of archaea) in the inoculum, and therefore their high abundance on the cathode resulted from selective enrichment. In contrast, bacterial communities on the cathode were more diverse, containing primarily δ-Proteobacteria (41% of bacteria). The lack of a consistent bacterial genus on the cathodes indicated that there was no similarly selective enrichment of bacteria on the cathode. These results suggest that the genus Methanobacterium was primarily responsible for methane production in MECs when cathodes lack efficient catalysts for hydrogen gas evolution. (Figure Presented).

Effect of methanogenic substrates on anaerobic oxidation of methane and sulfate reduction by an anaerobic methanotrophic enrichment.

KAUST Repository

Meulepas, Roel J W; Jagersma, Christian G; Khadem, Ahmad F; Stams, Alfons J M; Lens, Piet N L

2010-01-01

Anaerobic oxidation of methane (AOM) coupled to sulfate reduction (SR) is assumed to be a syntrophic process, in which methanotrophic archaea produce an interspecies electron carrier (IEC), which is subsequently utilized by sulfate-reducing bacteria

Iron oxides alter methanogenic pathways of acetate in production water of high-temperature petroleum reservoir.

Science.gov (United States)

Pan, Pan; Hong, Bo; Mbadinga, Serge Maurice; Wang, Li-Ying; Liu, Jin-Feng; Yang, Shi-Zhong; Gu, Ji-Dong; Mu, Bo-Zhong

2017-09-01

Acetate is a key intermediate in anaerobic crude oil biodegradation and also a precursor for methanogenesis in petroleum reservoirs. The impact of iron oxides, viz. β-FeOOH (akaganéite) and magnetite (Fe 3 O 4 ), on the methanogenic acetate metabolism in production water of a high-temperature petroleum reservoir was investigated. Methane production was observed in all the treatments amended with acetate. In the microcosms amended with acetate solely about 30% of the acetate utilized was converted to methane, whereas methane production was stimulated in the presence of magnetite (Fe 3 O 4 ) resulting in a 48.34% conversion to methane. Methane production in acetate-amended, β-FeOOH (akaganéite)-supplemented microcosms was much faster and acetate consumption was greatly improved compared to the other conditions in which the stoichiometric expected amounts of methane were not produced. Microbial community analysis showed that Thermacetogenium spp. (known syntrophic acetate oxidizers) and hydrogenotrophic methanogens closely related to Methanothermobacter spp. were enriched in acetate and acetate/magnetite (Fe 3 O 4 ) microcosms suggesting that methanogenic acetate metabolism was through hydrogenotrophic methanogenesis fueled by syntrophic acetate oxidizers. The acetate/β-FeOOH (akaganéite) microcosms, however, differed by the dominance of archaea closely related to the acetoclastic Methanosaeta thermophila. These observations suggest that supplementation of β-FeOOH (akaganéite) accelerated the production of methane further, driven the alteration of the methanogenic community, and changed the pathway of acetate methanogenesis from hydrogenotrophic methanogenesis fueled by syntrophic acetate oxidizers to acetoclastic.

Enhancement of anaerobic acidogenesis by integrating an electrochemical system into an acidogenic reactor: effect of hydraulic retention times (HRT) and role of bacteria and acidophilic methanogenic Archaea.

Science.gov (United States)

Zhang, Jingxin; Zhang, Yaobin; Quan, Xie; Chen, Shuo

2015-03-01

In this study, an acidogenic reactor packed with a pair of Fe-carbon electrodes (R1) was developed to enhance anaerobic acidogenesis of organic wastewater at short hydraulic retention times. The results indicated that the acidogenic efficiency was improved by settling a bio-electrochemical system. When hydraulic retention times decreased from 12 to 3h, R1 showed 18.9% more chemical oxygen demand removal and 13.8% more acidification efficiency. After cutting off the voltage of R1, the COD removal decreased by about 5%. Coupling of Fe(2+) leaching and electric field accelerated the hydrolysis of polysaccharide, relieving its accumulation in the sludge phase. Several acidophilic methanogenic Archaea such as Methanosarcina sp. were enriched in R1, which was favorable for consuming organic acids and preventing excessive pH decline. Thus, the developed acidogenic reactor with Fe-carbon electrodes is expected to be potentially effective and useful for wastewater treatment. Copyright © 2014 Elsevier Ltd. All rights reserved.

Bioaugmentation of an acetate-oxidising anaerobic consortium in up-flow sludge blanket reactor subjected to high ammonia loads

DEFF Research Database (Denmark)

Fotidis, Ioannis; Karakashev, Dimitar Borisov; Angelidaki, Irini

Ammonia is the major inhibitor of anaerobic digestion (AD) process leading to suboptimal utilisation of the biogas potential of the feedstocks and causing economical losses to the biogas plants. However, ammonia is mainly inhibiting the aceticlastic methanogens, while the hydrogenotrophic...... was tested. The co-cultivation in fed-batch of a fast-growing hydrogenotrophic methanogen (i.e. Methanoculleus bourgensis) with the SAO culture was also investigated. Results obtained clearly demonstrated that bioaugmentation of SAO culture in a UASB reactor was not possible most probably due to the slow...

Anaerobic co-digestion of food waste and landfill leachate in single-phase batch reactors

International Nuclear Information System (INIS)

Liao, Xiaofeng; Zhu, Shuangyan; Zhong, Delai; Zhu, Jingping; Liao, Li

2014-01-01

Highlights: • Anaerobic co-digestion strategy for food waste treatment at OLR 41.8 g VS/L. • A certain amount of raw leachate effectively relieved acidic inhibition. • The study showed that food waste was completely degraded. - Abstract: In order to investigate the effect of raw leachate on anaerobic digestion of food waste, co-digestions of food waste with raw leachate were carried out. A series of single-phase batch mesophilic (35 ± 1 °C) anaerobic digestions were performed at a food waste concentration of 41.8 g VS/L. The results showed that inhibition of biogas production by volatile fatty acids (VFA) occurred without raw leachate addition. A certain amount of raw leachate in the reactors effectively relieved acidic inhibition caused by VFA accumulation, and the system maintained stable with methane yield of 369–466 mL/g VS. Total ammonia nitrogen introduced into the digestion systems with initial 2000–3000 mgNH 4 –N/L not only replenished nitrogen for bacterial growth, but also formed a buffer system with VFA to maintain a delicate biochemical balance between the acidogenic and methanogenic microorganisms. UV spectroscopy and fluorescence excitation–emission matrix spectroscopy data showed that food waste was completely degraded. We concluded that using raw leachate for supplement water addition and pH modifier on anaerobic digestion of food waste was effective. An appropriate fraction of leachate could stimulate methanogenic activity and enhance biogas production

Anaerobic co-digestion of food waste and landfill leachate in single-phase batch reactors

Energy Technology Data Exchange (ETDEWEB)

Liao, Xiaofeng; Zhu, Shuangyan; Zhong, Delai; Zhu, Jingping, E-mail: jpzhuhust@163.com; Liao, Li, E-mail: liaoli2003@126.com

2014-11-15

Highlights: • Anaerobic co-digestion strategy for food waste treatment at OLR 41.8 g VS/L. • A certain amount of raw leachate effectively relieved acidic inhibition. • The study showed that food waste was completely degraded. - Abstract: In order to investigate the effect of raw leachate on anaerobic digestion of food waste, co-digestions of food waste with raw leachate were carried out. A series of single-phase batch mesophilic (35 ± 1 °C) anaerobic digestions were performed at a food waste concentration of 41.8 g VS/L. The results showed that inhibition of biogas production by volatile fatty acids (VFA) occurred without raw leachate addition. A certain amount of raw leachate in the reactors effectively relieved acidic inhibition caused by VFA accumulation, and the system maintained stable with methane yield of 369–466 mL/g VS. Total ammonia nitrogen introduced into the digestion systems with initial 2000–3000 mgNH{sub 4}–N/L not only replenished nitrogen for bacterial growth, but also formed a buffer system with VFA to maintain a delicate biochemical balance between the acidogenic and methanogenic microorganisms. UV spectroscopy and fluorescence excitation–emission matrix spectroscopy data showed that food waste was completely degraded. We concluded that using raw leachate for supplement water addition and pH modifier on anaerobic digestion of food waste was effective. An appropriate fraction of leachate could stimulate methanogenic activity and enhance biogas production.

Anaerobic degradation of the various fractions of slaughterhouse wastewater

Energy Technology Data Exchange (ETDEWEB)

Sayed, S.; Zanden, J. van der; Wijffels, R.; Lettinga, G.

1988-01-01

The main objectives of the present investigations were to determine the maximum extent of anaerobic biological degradation of the soluble, colloidal and Coarse Suspended Solids fractions of slaughterhouse wastewater, in order to elucidate the mechanisms involved in the removal of these organic pollutant fractions and to determine the rate-limiting steps in the degradation of each fraction of the wastewater, and to assess the effects of the fractions on the methanogenic activity of the sludge.

Anaerobic biotransformation of estrogens

International Nuclear Information System (INIS)

Czajka, Cynthia P.; Londry, Kathleen L.

2006-01-01

Estrogens are important environmental contaminants that disrupt endocrine systems and feminize male fish. We investigated the potential for anaerobic biodegradation of the estrogens 17-α-ethynylestradiol (EE2) and 17-β-estradiol (E2) in order to understand their fate in aquatic and terrestrial environments. Cultures were established using lake water and sediment under methanogenic, sulfate-, iron-, and nitrate-reducing conditions. Anaerobic degradation of EE2 (added at 5 mg/L) was not observed in multiple trials over long incubation periods (over three years). E2 (added at 5 mg/L) was transformed to estrone (E1) under all four anaerobic conditions (99-176 μg L -1 day -1 ), but the extent of conversion was different for each electron acceptor. The oxidation of E2 to E1 was not inhibited by E1. Under some conditions, reversible inter-conversion of E2 and E1 was observed, and the final steady state concentration of E2 depended on the electron-accepting condition but was independent of the total amount of estrogens added. In addition, racemization occurred and E1 was also transformed to 17-α-estradiol under all but nitrate-reducing conditions. Although E2 could be readily transformed to E1 and in many cases 17-α-estradiol under anaerobic conditions, the complete degradation of estrogens under these conditions was minimal, suggesting that they would accumulate in anoxic environments

Transformation of trinitrotoluene to triaminotoluene by mixed cultures incubated under methanogenic conditions

Energy Technology Data Exchange (ETDEWEB)

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.

Starch-degrading enzymes from anaerobic non-clostridial bacteria

Energy Technology Data Exchange (ETDEWEB)

Weber, H; Schepers, H J; Troesch, W [Fraunhofer-Institut fuer Grenzflaechen- und Bioverfahrenstechnik (IGB), Stuttgart (Germany, F.R.)

1990-08-01

A number of meso- and thermophilic anaerobic starch-degrading non-spore-forming bacteria have been isolated. All the isolates belonging to different genera are strictly anaerobic, as indicated by a catalase-negative reaction, and produce soluble starch-degrading enzymes. Compared to enzymes of aerobic bacteria, those of anaerobic origin mainly show low molecular mass of about 25 000 daltons. Some of the enzymes may have useful applications in the starch industry because of their unusual product pattern, yielding maltotetraose as the main hydrolysis product. (orig.).

Biotransformation potential of phytosterols under anoxic and anaerobic conditions.

Science.gov (United States)

Dykstra, C M; Giles, H D; Banerjee, S; Pavlostathis, S G

2014-01-01

The biotransformation potential of three phytosterols (campesterol, stigmasterol and β-sitosterol) under denitrifying, sulfate-reducing and fermentative/methanogenic conditions was assessed. Using a group contribution method, the standard Gibbs free energy of phytosterols was calculated and used to perform theoretical energetic calculations. The oxidation of phytosterols under aerobic, nitrate-reducing, sulfate-reducing and methanogenic conditions was determined to be energetically feasible. However, using semi-continuously fed cultures maintained at 20-22 °C over 16 weekly feeding cycles (112 days; retention time, 21 days), phytosterol removal was observed under nitrate-reducing and sulfate-reducing conditions, but not under fermentative/methanogenic conditions. Under sulfate-reducing conditions, stigmast-4-en-3-one was identified as an intermediate of phytosterol biotransformation, a reaction more likely carried out by dehydrogenases/isomerases, previously reported to act on cholesterol under both oxic and anoxic (denitrifying) conditions. Further study of the biotransformation of phytosterols under anoxic/anaerobic conditions is necessary to delineate the factors and conditions leading to enhanced phytosterol biodegradation and the development of effective biological treatment systems for the removal of phytosterols from pulp and paper wastewaters and other phytosterol-bearing waste streams.

Microbial Internal Storage Alters the Carbon Transformation in Dynamic Anaerobic Fermentation.

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Ni, Bing-Jie; Batstone, Damien; Zhao, Bai-Hang; Yu, Han-Qing

2015-08-04

Microbial internal storage processes have been demonstrated to occur and play an important role in activated sludge systems under both aerobic and anoxic conditions when operating under dynamic conditions. High-rate anaerobic reactors are often operated at a high volumetric organic loading and a relatively dynamic profile, with large amounts of fermentable substrates. These dynamic operating conditions and high catabolic energy availability might also facilitate the formation of internal storage polymers by anaerobic microorganisms. However, so far information about storage under anaerobic conditions (e.g., anaerobic fermentation) as well as its consideration in anaerobic process modeling (e.g., IWA Anaerobic Digestion Model No. 1, ADM1) is still sparse. In this work, the accumulation of storage polymers during anaerobic fermentation was evaluated by batch experiments using anaerobic methanogenic sludge and based on mass balance analysis of carbon transformation. A new mathematical model was developed to describe microbial storage in anaerobic systems. The model was calibrated and validated by using independent data sets from two different anaerobic systems, with significant storage observed, and effectively simulated in both systems. The inclusion of the new anaerobic storage processes in the developed model allows for more successful simulation of transients due to lower accumulation of volatile fatty acids (correction for the overestimation of volatile fatty acids), which mitigates pH fluctuations. Current models such as the ADM1 cannot effectively simulate these dynamics due to a lack of anaerobic storage mechanisms.

Influence of Martian regolith analogs on the activity and growth of methanogenic archaea, with special regard to long-term desiccation

Directory of Open Access Journals (Sweden)

Janosch eSchirmack

2015-03-01

Full Text Available Methanogenic archaea have been studied as model organisms for possible life on Mars for several reasons: they can grow lithoautotrophically by using hydrogen and carbon dioxide as energy and carbon sources, respectively; they are anaerobes; and they evolved at a time when conditions on early Earth are believed to have looked similar to those of early Mars. As Mars is currently dry and cold and as water might be available only at certain time intervals, any organism living on this planet would need to cope with desiccation. On Earth there are several regions with low water availability as well, e.g. permafrost environments, desert soils and salt pans. Here, we present the results of a set of experiments investigating the influence of different Martian regolith analogs on the metabolic activity and growth of three methanogenic strains exposed to culture conditions as well as long-term desiccation. In most cases, concentrations below 1 %wt of regolith in the media resulted in an increase of methane production rates, whereas higher concentrations decreased the rates, thus prolonging the lag phase. Further experiments showed that methanogenic archaea are capable of producing methane when incubated on a water-saturated sedimentary matrix of regolith lacking nutrients. Survival of methanogens under these conditions was analyzed with a 400 day desiccation experiment in the presence of regolith analogs. All tested strains of methanogens survived the desiccation period as it was determined through reincubation on fresh medium and via qPCR following propidium monoazide treatment to identify viable cells. The survival of long-term desiccation and the ability of active metabolism on water-saturated MRAs strengthens the possibility of methanogenic archaea or physiologically similar organisms to exist in environmental niches on Mars. The best results were achieved in presence of a phyllosilicate, which provides insights of possible positive effects in habitats

Conversion of methanogenic substrates in anaerobic reactors : metals, mass transport, and toxicity

NARCIS (Netherlands)

Gonzalez-Gil, G.

2000-01-01

The EGSB systems represents an attractive option to extend further the use of anaerobic technology for wastewater treatment, particularly with respect to waste streams originating from chemical industries. Frequently chemical waste streams are unbalanced with respect to nutrients and/or

Effect of Nitrooxy Compounds with Different Molecular Structures on the Rumen Methanogenesis, Metabolic Profile, and Methanogenic Community.

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Jin, Wei; Meng, Zhenxiang; Wang, Jing; Cheng, Yanfen; Zhu, Weiyun

2017-08-01

Rumen in vitro fermentation was used to evaluate the capacity of nitrooxy compounds to mitigate rumen methane production. The following three nitrooxy compounds, each with different molecular structures, were evaluated: 2,2-dimethyl-3-(nitrooxy) propanoic (DNP), N-[2-(Nitrooxy)ethyl]-3-pyridinecarboxamide (NPD), and nitroglycerin (NG). All three compounds substantially decreased the total gas production, methane production, and the acetate:propionate ratio, while increasing hydrogen production. The growth of methanogens was specifically inhibited by all three compounds, without affecting the abundance of bacteria, anaerobic fungi, or protozoa. However, inhibition of methanogenesis required a much higher dose of DNP when compared to NPD or NG. Further investigations were conducted on NG to determine its effects on the methanogenic community. NG reduced the relative abundance of Methanomassiliicoccales, while increasing the relative abundance of Methanobrevibacter and Methanosphaera. Overall, the results suggested that all three of these nitrooxy compounds could specifically inhibit rumen methanogenesis, but NPD and NG were much more efficient than DNP at rumen methane mitigation.

[Role of obligate anaerobes in infections in hospitalized patients and therapeutic options].

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Wróblewska, Marta; Rokosz, Alicja; Sawicka-Grzelak, Anna; Kot, Katarzyna; Luczak, Mirosław

2005-01-01

Monitoring of prevalence and antibiotic susceptibility of strictly anaerobic bacteria, causing infections in hospitalized patients, constitutes a part of a program for prudent use of antibiotics. The aim of the study was to assess prevalence of strictly anaerobic bacteria in patients hospitalized in a tertiary care hospital in 2001-2002 with reference to empiric antibiotic therapy. The most common gram-positive bacteria were Clostridium difficile--27.7%, Peptostreptococcus spp. and Peptoniphilus asaccharolyticus--21.9% and Actinomyces spp.--11.1%. There was an increase in the number of stool samples positive for C. difficile toxins A and B from 39.4% in 2001 to 59.0% in 2002. The results of susceptibility testing of gram-positive isolates showed the highest percentages of strains susceptible to piperacilin/tazobactam--99.6%, ticarcillin/clavulanate--98.5%, imipenem--98.5%, amoxicillin/clavulanate--97.4% and piperacillin--97.4%. The most prevalent gram-negative anaerobes were strains of Bacteroides spp.--43.1%, Prevotella spp.--35.8% and Fusobacterium spp.--11.0%. All tested strains of gram-negative bacteria were susceptible to metronidazole, piperacilin/tazobactam, ticarcillin/clavulanate and imipenem. In the analyzed population beta-lactam antibiotics with beta-lactamase inhibitors, carbapenems and metronidazole may be used in empiric therapy of infections caused by strictly anaerobic bacteria.

Kraft pulp and paper mill wastewater treatment using fixed bed anaerobic reactors

International Nuclear Information System (INIS)

Damianovic, M. H. R. Z; Ruas, D.; Pires, E. C.; Foresti, E.

2009-01-01

The effluents of pulp mills contain a myriad of toxic compounds, biodegradable organic matter and sulfur compounds. to decrease the amount of fresh water required for pulp and paper production closed circuits are in use, however, higher concentrations of slat, as oxidized sulfur compounds, are encountered in the wastewaters. energy costs and new environmental concerns are motivating the use of anaerobic pretreatment as a way to decrease energy expenditure in the treatment plant together with lower sludge production. In anaerobic environment, the organic matter removal can follow methanogenic or sulfidogenic paths and with the latter simultaneous reduction of the oxidized sulfur compounds also occurs. (Author)

Increased methane production in cyanobacteria and methanogenic microbe co-cultures.

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Yeung, Tracey; Kwan, Matthew; Adler, Lewis; Mills, Toby J; Neilan, Brett A; Conibeer, Gavin; Patterson, Robert

2017-11-01

A novel light-to-bioenergy system produced 3.5 times the baseline methane output using a co-culture of cyanobacteria (Oscillatoria sp.) and a methanogenic microbial community. Analysis of micronutrients in the system during the growth phase indicated that cobalt, iron, nickel and zinc were not appreciably consumed. The stable consumption and return of macronutrients calcium and magnesium were also observed. Essential macronutrients nitrogen, in the form of nitrate, and phosphorus showed no cycling during the growth phase and were depleted at rates of 0.35mg/L/day and 0.40µg/L/day, respectively. Biofilm formation increased the resilience of biomass to bacterial degradation in an anaerobic digester, as shown by viability assays of cyanobacterial biofilms in the co-culture. Copyright © 2017 Elsevier Ltd. All rights reserved.

Metabolism of reduced methylated sulfur compounds in anaerobic sediments and by a pure culture of an estuarine methanogen

International Nuclear Information System (INIS)

Kiene, R.P.; Oremland, R.S.; Catena, A.; Miller, L.G.; Capone, A.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 ∼2-3=M levels as [ 14 C]DMS, metabolism by sediments resulted in a 14 CH 4 / 14 CO 2 ratio of only 0.06. Addition of molybdate increased the ratio of 1.8, while 2-bromoethanesulfonic acid decreased it to 0, but did not block 14 CO 2 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 [ 14 C]-DMS to yield a 14 CH 4 / 14 CO 2 ratio of ∼ 2.8

Simultaneous Cr(VI) bio-reduction and methane production by anaerobic granular sludge.

Science.gov (United States)

Hu, Qian; Sun, Jiaji; Sun, Dezhi; Tian, Lan; Ji, Yanan; Qiu, Bin

2018-08-01

Wastewater containing toxic hexavalent chromium (Cr(VI)) were treated with well-organized anaerobic granular sludge in this study. Results showed that the anaerobic granular sludge rapidly removed Cr(VI), and 2000 µg·L -1 Cr(VI) was completely eliminated within 6 min, which was much faster than the reported duration of removal by reported artificial materials. Sucrose added as a carbon source acted as an initial electron donor to reduce Cr(VI) to Cr(III). This process was considered as the main mechanism of Cr(VI) removal. Methane production by anaerobic granular sludge was improved by the addition of Cr(VI) at a concentration lower than 500 µg·L -1 . Anaerobic granular sludge had a well-organized structure, which presented good resistance against toxic Cr(VI). Trichoccus accelerated the degradation of organic substances to generate acetates with a low Cr(VI) concentration, thereby enhancing methane production by acetotrophic methanogens. Copyright © 2018 Elsevier Ltd. All rights reserved.

Relating Anaerobic Digestion Microbial Community and Process Function.

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Venkiteshwaran, Kaushik; Bocher, Benjamin; Maki, James; Zitomer, Daniel

2015-01-01

Anaerobic digestion (AD) involves a consortium of microorganisms that convert substrates into biogas containing methane for renewable energy. The technology has suffered from the perception of being periodically unstable due to limited understanding of the relationship between microbial community structure and function. The emphasis of this review is to describe microbial communities in digesters and quantitative and qualitative relationships between community structure and digester function. Progress has been made in the past few decades to identify key microorganisms influencing AD. Yet, more work is required to realize robust, quantitative relationships between microbial community structure and functions such as methane production rate and resilience after perturbations. Other promising areas of research for improved AD may include methods to increase/control (1) hydrolysis rate, (2) direct interspecies electron transfer to methanogens, (3) community structure-function relationships of methanogens, (4) methanogenesis via acetate oxidation, and (5) bioaugmentation to study community-activity relationships or improve engineered bioprocesses.

Analysis of alkane-dependent methanogenic community derived from production water of a high-temperature petroleum reservoir

Energy Technology Data Exchange (ETDEWEB)

Mbadinga, Serge Maurice; Li, Kai-Ping; Zhou, Lei; Wang, Li-Ying; Yang, Shi-Zhong; Liu, Jin-Feng; Mu, Bo-Zhong [East China Univ. of Science and Technology, Shanghai (China). State Key Lab. of Bioreactor Engineering and Inst. of Applied Chemistry; Gu, Ji-Dong [Hong Kong Univ. (China). School of Biological Sciences

2012-10-15

Microbial assemblage in an n-alkanes-dependent thermophilic methanogenic enrichment cultures derived from production waters of a high-temperature petroleum reservoir was investigated in this study. Substantially higher amounts of methane were generated from the enrichment cultures incubated at 55 C for 528 days with a mixture of long-chain n-alkanes (C{sub 15}-C{sub 20}). Stoichiometric estimation showed that alkanes-dependent methanogenesis accounted for about 19.8% of the total amount of methane expected. Hydrogen was occasionally detected together with methane in the gas phase of the cultures. Chemical analysis of the liquid cultures resulted only in low concentrations of acetate and formate. Phylogenetic analysis of the enrichment revealed the presence of several bacterial taxa related to Firmicutes, Thermodesulfobiaceae, Thermotogaceae, Nitrospiraceae, Dictyoglomaceae, Candidate division OP8 and others without close cultured representatives, and Archaea predominantly related to uncultured members in the order Archaeoglobales and CO{sub 2}-reducing methanogens. Screening of genomic DNA retrieved from the alkanes-amended enrichment cultures also suggested the presence of new alkylsuccinate synthase alpha-subunit (assA) homologues. These findings suggest the presence of poorly characterized (putative) anaerobic n-alkanes degraders in the thermophilic methanogenic enrichment cultures. Our results indicate that methanogenesis of alkanes under thermophilic condition is likely to proceed via syntrophic acetate and/or formate oxidation linked with hydrogenotrophic methanogenesis. (orig.)

Thermophilic two-phase anaerobic digestion using an innovative fixed-bed reactor for enhanced organic matter removal and bioenergy recovery from sugarcane vinasse

International Nuclear Information System (INIS)

Fuess, Lucas Tadeu; Kiyuna, Luma Sayuri Mazine; Ferraz, Antônio Djalma Nunes; Persinoti, Gabriela Felix

2017-01-01

Highlights: • An innovative fixed-film anaerobic reactor was applied to sugarcane vinasse. • Stable operation was observed for OLRs as high as 30 kg COD m"−"3 day"−"1. • Propionate buildup did not impact the stability of the structured-bed reactor. • Enhanced bioenergy recovery was estimated from biodigestion with phase separation. • Energy extraction was over 20% higher compared to single-phase systems. - Abstract: This study considered the application of anaerobic digestion (AD) with phase separation combined with the use of an anaerobic structured-bed reactor (ASTBR) as the methanogenic phase for the treatment of sugarcane vinasse, a high-strength wastewater resulting from ethanol production. Two combined thermophilic acidogenic-methanogenic systems formed by one single acidogenic reactor followed by two methanogenic reactors operated in parallel were compared, namely, a conventional UASB reactor and an upflow ASTBR reactor. Increasing organic loading rate (OLR) conditions (15–30 kg COD m"−"3 d"−"1) were applied to the methanogenic reactors. The results highlighted the feasibility of applying the ASTBR to vinasse, indicating a global COD removal higher than 80%. The ASTBR exhibited a stable long-term operation (240 days), even for OLR values as high as 30 kg COD m"−"3 d"−"1. The application of similar conditions to the UASB reactor indicated severe performance losses, leading to the accumulation of acids for every increase in the OLR. An energetic potential of 181.5 MJ for each cubic meter of vinasse was estimated from both hydrogen and methane. The provision of bicarbonate alkalinity proved to be a key factor in obtaining stable performance, offsetting the limitations of relatively low hydraulic retention times (<24 h).

Influence of water solubility, side chain degradability and side chain configuration on the degradation of phthalic acid esters under methanogenic conditions

Energy Technology Data Exchange (ETDEWEB)

Alnervik, M.

1996-12-31

Water solubility and degradability of side chains estrifying phthalic acid are factors possible to influence the degradation of phthalic acid esters (PAEs). To investigate the importance of these factors degradation of butyl 2-ethylhexyl phthalate (BEHP), bis(2-ethylhexyl) phthalate (DEHP), dibutyl phthalate (DBP), dihexyl phthalate (DHP), dioctyl phthalate (DOP) and didecyl phthalate (DDP) were examined under methanogenic conditions as well as was the degradability of the alcohols estrifying these PAEs. We also investigated if the degradation of resistant PAEs could be stimulated by the addition of a degradable PAE. Synthesis of degradation intermediates and two methods for PAE analyses are presented. The investigation showed that all alcohols were degraded to methane and carbon dioxide and that the degradation of PAE occurred in incubations amended with BBP, BEHP, DHP and DBP, whilst DEHP, DOP and DDP were unaffected throughout the experimental period. BBP added to incubations with DEHP, could not stimulate DEHP degradation. In conclusion, the degradability of alcohols estrifying phthalic acid in this study does not affect the anaerobic degradability of PAEs. Water solubility of a PAE can not be rejected as a factor limiting phthalate degradation under methanogenic conditions. Anaerobic degradation of persistent PAEs can not be stimulated by mixing it with a degradable phthalate. 23 refs, 11 figs, 2 tabs

The aerobic activity of metronidazole against anaerobic bacteria.

Science.gov (United States)

Dione, Niokhor; Khelaifia, Saber; Lagier, Jean-Christophe; Raoult, Didier

2015-05-01

Recently, the aerobic growth of strictly anaerobic bacteria was demonstrated using antioxidants. Metronidazole is frequently used to treat infections caused by anaerobic bacteria; however, to date its antibacterial activity was only tested in anaerobic conditions. Here we aerobically tested using antioxidants the in vitro activities of metronidazole, gentamicin, doxycycline and imipenem against 10 common anaerobic and aerobic bacteria. In vitro susceptibility testing was performed by the disk diffusion method, and minimum inhibitory concentrations (MICs) were determined by Etest. Aerobic culture of the bacteria was performed at 37°C using Schaedler agar medium supplemented with 1mg/mL ascorbic acid and 0.1mg/mL glutathione; the pH was adjusted to 7.2 by 10M KOH. Growth of anaerobic bacteria cultured aerobically using antioxidants was inhibited by metronidazole after 72h of incubation at 37°C, with a mean inhibition diameter of 37.76mm and an MIC of 1μg/mL; however, strains remained non-sensitive to gentamicin. No growth inhibition of aerobic bacteria was observed after 24h of incubation at 37°C with metronidazole; however, inhibition was observed with doxycycline and imipenem used as controls. These results indicate that bacterial sensitivity to metronidazole is not related to the oxygen tension but is a result of the sensitivity of the micro-organism. In future, both culture and antibiotic susceptibility testing of strictly anaerobic bacteria will be performed in an aerobic atmosphere using antioxidants in clinical microbiology laboratories. Copyright © 2015 Elsevier B.V. and the International Society of Chemotherapy. All rights reserved.

Effect of nitrogen fertilizer and/or rice straw amendment on methanogenic archaeal communities and methane production from a rice paddy soil.

Science.gov (United States)

Bao, Qiongli; Huang, Yizong; Wang, Fenghua; Nie, Sanan; Nicol, Graeme W; Yao, Huaiying; Ding, Longjun

2016-07-01

Nitrogen fertilization and returning straw to paddy soil are important factors that regulate CH4 production. To evaluate the effect of rice straw and/or nitrate amendment on methanogens, a paddy soil was anaerobically incubated for 40 days. The results indicated that while straw addition increased CH4 production and the abundances of mcrA genes and their transcripts, nitrate amendment showed inhibitory effects on them. The terminal restriction fragment length polymorphism (T-RFLP) analysis based on mcrA gene revealed that straw addition obviously changed methanogenic community structure. Based on mcrA gene level, straw-alone addition stimulated Methanosarcinaceaes at the early stage of incubation (first 11 days), but nitrate showed inhibitory effect. The relative abundance of Methanobacteriaceae was also stimulated by straw addition during the first 11 days. Furthermore, Methanosaetaceae were enriched by nitrate-alone addition after 11 days, while Methanocellaceae were enriched by nitrate addition especially within the first 5 days. The transcriptional methanogenic community indicated more dynamic and complicated responses to straw and/or nitrate addition. Based on mcrA transcript level, nitrate addition alone resulted in the increase of Methanocellaceae and the shift from Methanosarcinaceae to Methanosaetaceae during the first 5 days of incubation. Straw treatments increased the relative abundance of Methanobacteriaceae after 11 days. These results demonstrate that nitrate addition influences methanogens which are transcriptionally and functionally active and can alleviate CH4 production associated with straw amendment in paddy soil incubations, presumably through competition for common substrates between nitrate-utilizing organisms and methanogens.

Toxicity in anaerobic digestion : with emphasis on the effect of ammonia, sulfide and long-chain fatty acids on methanogenesis

NARCIS (Netherlands)

Koster, I.W.

1989-01-01

The dissertation concerns the problem of toxicity in anaerobic digestion, which to a large extent is the problem of inhibition of methanogenic conversions by chemical compounds. The dissertation begins with an extensive literature review in which the