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Sample records for methanogenic bioreactor packed

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

    PubMed

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

    2010-05-01

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

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

    PubMed

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

    2010-05-01

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

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

    PubMed

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

    2015-02-01

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

  4. LEACHATE RECIRCULATION, METHANOGENS AND METAL CONCENTRATIONS IN BIOREACTOR LANDFILLS

    EPA Science Inventory

    The idea of operating landfills as bioreactors has received a lot of attention owing to many of the economic and waste treatment benefits. Portions of the Outer Loop landfill in Louisville, KY, owned and operated by WMI, Inc., are currently being used to test two different decom...

  5. Modeling anaerobic bioreactor landfills in methanogenic phase: long term and short term behaviors.

    PubMed

    Gholamifard, Shabnam; Eymard, Robert; Duquennoi, Christian

    2008-12-01

    We have developed a mathematical model to simulate the behavior of real bioreactor landfills in the anaerobic methanogenic phase. This coupled model is composed of a two-phase flow and a biological model based on Darcy's law and Monod's model, respectively. This model considers bacterial activity and biological behavior as a function of temperature and makes it possible to study the thermo-biological behavior of bioreactor landfills with temperature changes. In this model we consider different effects of saturation on solid waste degradation. These effects consist of increasing hydrolysis with saturation and also decreasing the concentration of volatile fatty acids (VFAs) and activating the methanogenic biomass. This paper presents first the mathematical coupled model and the numerical methods used to solve the conservation equations. The numerical model is then used to simulate two bioreactor landfills. This paper presents the results of long and short (with leachate recirculation) term numerical simulations comparing them with site results. Finally results as well as advantages and drawbacks of the model are discussed. The results show that the mathematical model is able to reproduce the hydro-thermo-biological behavior of a bioreactor landfill in different conditions, with and without leachate recirculation, and leads to a better understanding of important thermal and biological parameters.

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

    PubMed

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

    2015-08-01

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

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

    PubMed

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

    2015-08-01

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

  8. A comparison of nanosilver and silver ion effects on bioreactor landfill operations and methanogenic population dynamics.

    PubMed

    Yang, Yu; Gajaraj, Shashikanth; Wall, Judy D; Hu, Zhiqiang

    2013-06-15

    Silver nanoparticles (AgNPs, nanosilver) and silver ions released from industry and various consumer products are eventually disposed in sanitary landfills. To compare the effects of these two forms of silver on landfill bioreactor operations with leachate recirculation, municipal solid waste (MSW) in six identical 9-L bioreactors was exposed to AgNPs (stabilized with 0.06% polyvinyl alcohol) or Ag(+) at a silver concentration of 10 mg/kg solids. The landfill anaerobic digestion was operated and monitored for more than 200 days. There was no significant difference in cumulative methane volume or methane production rate between the groups of control and 10 mg/kg Ag(+). However, MSW treated with 10 mg/kg AgNPs resulted in a reduced methane production (by up to 80%) and accumulation of volatile fatty acids in the leachates. This was accompanied by higher leachate Ag concentrations (an average of 3.7 ± 0.3 mg/L) after day 132 as compared to those in the groups of control and 10 mg/kg Ag(+) at 0.7 ± 0.4 and 1.1 ± 0.3 mg/L, respectively. Quantitative PCR targeting 16S rRNA genes of methanogens indicated reduced methanogenic growth in the bioreactors exposed to nanosilver. The peak values of total methanogens in leachates were (1.18 ± 0.09) × 10(10), (4.57 ± 2.67) × 10(10) and (7.72 ± 0.78) × 10(8) (cells/mL) for the groups of control, Ag(+) and AgNPs, respectively. The results suggest that silver ions have minimal impact on landfill methane production at the concentration of 10 mg/kg. However, nanosilver inhibits methanogenesis and is more toxic than its counterpart, likely due to slow and long-term Ag(+) release from nanosilver dissolution yielding more bioavailability in landfill leachates.

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

    PubMed Central

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

    2011-01-01

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

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

    NASA Astrophysics Data System (ADS)

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

    2013-12-01

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

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

    PubMed

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

    2015-11-01

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

  12. Efficacy of Aspergillus sp. for degradation of chlorpyrifos in batch and continuous aerated packed bed bioreactors.

    PubMed

    Yadav, Maya; Srivastva, Navnita; Shukla, Awadhesh Kumar; Singh, Ram Sharan; Upadhyay, Siddh Nath; Dubey, Suresh Kumar

    2015-01-01

    Aerobic biodegradation of chlorpyrifos (CP) by Aspergillus sp. was investigated in batch and continuous packed bed bioreactors. The optimal process parameters for achieving the maximum removal efficiency (RE), determined using a batch bioreactor packed with polyurethane foam pieces, were inoculum level: 2.5 mg (wet weight) mL(-1), pH 7.0, temperature 28 °C, DO 5.8 mg L(-1), and CP concentration 300 mg L(-1). The continuous packed bed bioreactor was operated at flow rates ranging from 10 to 40 mL h(-1) while keeping other parameters at their optimal level. Steady-state CP removal efficiencies greater than 85 % were obtained up to the inlet loading of 180 mg L(-1) d(-1). The continuous bioreactor behaved as a plug flow unit and was able to stabilize quickly after perturbation in the inlet loading.

  13. Packed Bed Bioreactor for the Isolation and Expansion of Placental-Derived Mesenchymal Stromal Cells

    PubMed Central

    Osiecki, Michael J.; Michl, Thomas D.; Kul Babur, Betul; Kabiri, Mahboubeh; Atkinson, Kerry; Lott, William B.; Griesser, Hans J.; Doran, Michael R.

    2015-01-01

    Large numbers of Mesenchymal stem/stromal cells (MSCs) are required for clinical relevant doses to treat a number of diseases. To economically manufacture these MSCs, an automated bioreactor system will be required. Herein we describe the development of a scalable closed-system, packed bed bioreactor suitable for large-scale MSCs expansion. The packed bed was formed from fused polystyrene pellets that were air plasma treated to endow them with a surface chemistry similar to traditional tissue culture plastic. The packed bed was encased within a gas permeable shell to decouple the medium nutrient supply and gas exchange. This enabled a significant reduction in medium flow rates, thus reducing shear and even facilitating single pass medium exchange. The system was optimised in a small-scale bioreactor format (160 cm2) with murine-derived green fluorescent protein-expressing MSCs, and then scaled-up to a 2800 cm2 format. We demonstrated that placental derived MSCs could be isolated directly within the bioreactor and subsequently expanded. Our results demonstrate that the closed system large-scale packed bed bioreactor is an effective and scalable tool for large-scale isolation and expansion of MSCs. PMID:26660475

  14. Packed Bed Bioreactor for the Isolation and Expansion of Placental-Derived Mesenchymal Stromal Cells.

    PubMed

    Osiecki, Michael J; Michl, Thomas D; Kul Babur, Betul; Kabiri, Mahboubeh; Atkinson, Kerry; Lott, William B; Griesser, Hans J; Doran, Michael R

    2015-01-01

    Large numbers of Mesenchymal stem/stromal cells (MSCs) are required for clinical relevant doses to treat a number of diseases. To economically manufacture these MSCs, an automated bioreactor system will be required. Herein we describe the development of a scalable closed-system, packed bed bioreactor suitable for large-scale MSCs expansion. The packed bed was formed from fused polystyrene pellets that were air plasma treated to endow them with a surface chemistry similar to traditional tissue culture plastic. The packed bed was encased within a gas permeable shell to decouple the medium nutrient supply and gas exchange. This enabled a significant reduction in medium flow rates, thus reducing shear and even facilitating single pass medium exchange. The system was optimised in a small-scale bioreactor format (160 cm2) with murine-derived green fluorescent protein-expressing MSCs, and then scaled-up to a 2800 cm2 format. We demonstrated that placental derived MSCs could be isolated directly within the bioreactor and subsequently expanded. Our results demonstrate that the closed system large-scale packed bed bioreactor is an effective and scalable tool for large-scale isolation and expansion of MSCs.

  15. Packed Bed Bioreactor for the Isolation and Expansion of Placental-Derived Mesenchymal Stromal Cells.

    PubMed

    Osiecki, Michael J; Michl, Thomas D; Kul Babur, Betul; Kabiri, Mahboubeh; Atkinson, Kerry; Lott, William B; Griesser, Hans J; Doran, Michael R

    2015-01-01

    Large numbers of Mesenchymal stem/stromal cells (MSCs) are required for clinical relevant doses to treat a number of diseases. To economically manufacture these MSCs, an automated bioreactor system will be required. Herein we describe the development of a scalable closed-system, packed bed bioreactor suitable for large-scale MSCs expansion. The packed bed was formed from fused polystyrene pellets that were air plasma treated to endow them with a surface chemistry similar to traditional tissue culture plastic. The packed bed was encased within a gas permeable shell to decouple the medium nutrient supply and gas exchange. This enabled a significant reduction in medium flow rates, thus reducing shear and even facilitating single pass medium exchange. The system was optimised in a small-scale bioreactor format (160 cm2) with murine-derived green fluorescent protein-expressing MSCs, and then scaled-up to a 2800 cm2 format. We demonstrated that placental derived MSCs could be isolated directly within the bioreactor and subsequently expanded. Our results demonstrate that the closed system large-scale packed bed bioreactor is an effective and scalable tool for large-scale isolation and expansion of MSCs. PMID:26660475

  16. Biodegradation of chlorpyrifos by Pseudomonas sp. in a continuous packed bed bioreactor.

    PubMed

    Yadav, Maya; Srivastva, Navnita; Singh, Ram Sharan; Upadhyay, Siddh Nath; Dubey, Suresh Kumar

    2014-08-01

    Biodegradation of chlorpyrifos (CP) by Pseudomonas (Iso 1) sp. was investigated in batch as well as continuous bioreactors packed with polyurethane foam pieces. The optimum process parameters for the maximum removal of CP, determined through batch experiments, were found to be: inoculum level, 300×10(6)CfumL(-1); CP concentration, 500mgL(-1); pH 7.5; temperature, 37°C and DO, 5.5mgL(-1). The continuous packed bed bioreactor was operated at various flow rates (10-40mLh(-1)) under the optimum conditions. The steady state CP removal efficiency of more than 91% was observed up to the inlet load of 300mgL(-1)d(-1). The bioreactor was sensitive to flow fluctuations but was able to recover its performance quickly and exhibited the normal plug-flow behavior. Accumulation of TCP (3,5,6-trichloro-2-pyridinol) affected the reactor performance.

  17. Continuous, packed-bed, enzymatic bioreactor production and stability of feruloyl soy glycerides

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The synthesis of feruloyl soy glycerides was demonstrated on a pilot-scale (1 metric ton/year) in a continuous, four-column series, packed-bed, enzymatic bioreactor. Ethyl ferulate and soybean oil were combined and converted at 3.5 kg/d over Candida antartica lipase B immobilized on an acrylic rein ...

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

    PubMed

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

    2009-08-01

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

  19. Production of rhEPO with a serum-free medium in the packed bed bioreactor.

    PubMed

    Deng, J; Yang, Q; Cheng, X; Li, L; Zhou, J

    1997-01-01

    Recombinant CHO (C2) cells producing human erythropoietin (rhEPO) were cultured with DMEM:F12 media containing 5% FBS for 8-10 days in a packed bed bioreactor, then rhEPO was produced with a serum-free medium (SFM-p) which was prepared in our laboratory. The SFM-p medium can support the growth of C2 cells and the production of rhEPO, and furthermore, it easily separates rhEPO from the culture supernatant. The cell culture in a packed bed bioreactor system using SFM-p was maintained in a stable condition for 20-25 days. The expression level of rhEPO was 12-28.4 mg/L. The bioreactor productivity was 71.0 mg/L.d and increased by 12-14 fold over that of the roller bottle. The glucose consumption rate was 21 g/L.d. At the end of 30 days of perfusion circulation, a final cell density of over 3.0 x 10(7)/ml of culture volume was achieved. Since the cells were entrapped in the polyester disk, the culture supernatant contained only a few detachment cells. Variations in lactate and ammonia production in the reactor were observed, and results showed that the productions of lactate and ammonia by the bioreactor were 3.5 g/L and 5 mmol/L, respectively, and did not affect the expression of interest protein. This experiment demonstrates that SFM-p is suitable for the growth and rhEPO production of recombinant C2 in the packed bed bioreactor.

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

    PubMed

    Morita, Masahiko; Sasaki, Kengo

    2012-05-01

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

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

    PubMed

    Morita, Masahiko; Sasaki, Kengo

    2012-05-01

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

  2. Continuous acetonitrile degradation in a packed-bed bioreactor.

    PubMed

    Manolov, Taras; Kristina, Håkansson; Benoit, Guieysse

    2005-02-01

    A 20-l packed-bed reactor filled with foamed glass beads was tested for the treatment of acetonitrile HPLC wastes. Aeration was provided by recirculating a portion of the reactor liquid phase through an aeration tank, where the dissolved oxygen concentration was kept at 6 mg/l. At a feeding rate of 0.77 g acetonitrile l(-1) reactor day(-1), 99% of the acetonitrile was removed; and 86% of the nitrogen present in acetonitrile was released as NH3, confirming that acetonitrile volatilization was not significant. Increasing the acetonitrile loading resulted in lower removal efficiencies, but a maximum removal capacity of 1.0 g acetonitrile l(-1) reactor day(-1) was achieved at a feeding rate of 1.6 g acetonitrile l(-1) reactor day(-1). The removal capacity of the system was well correlated with the oxygenation capacity, showing that acetonitrile removal was likely to be limited by oxygen supply. Microbial characterization of the biofilm resulted in the isolation of a Comamonas sp. able to mineralize acetonitrile as sole carbon, nitrogen and energy source. This organism was closely related to C. testosteroni (91.2%) and might represent a new species in the Comamonas genus. This study confirms the potential of packed-bed reactors for the treatment of a concentrated mixture of volatile pollutants.

  3. Degradation of carbon tetrachloride in a semi-continuous packed-bed bioreactor

    SciTech Connect

    Levinson, W.E.; Korus, R.A.; Crawford, R.L.

    1995-12-31

    Pseudomonas sp. KC degrades carbon tetrachloride (CT) when grown under iron limited conditions. A packed-bed bioreactor was constructed out of glass and Teflon components and contained glass beads as a packing material. Total void volume of the reactor, including the pump reservoir, was approximately 300 mL. Media was recirculated through the reactor with a Teflon diaphragm pump and contained nitrate as terminal electron acceptor. After inoculation with KC and biofilm growth, the pump reservoir was replaced with fresh media and 3.0 mg/L CT was introduced to the reactor. When the initial CT had been degraded, 10% of the reactor volume was replaced with new media and CT was reintroduced. This process was repeated a second time. The initial degradation rate of the reactor increased from 75 {mu}g CT L{sup -1} hr{sup -1} for the first feeding to 109 {mu}g L{sup -1} hr{sup -1} for the third. A second reactor was packed with a Celite (diatomaceous earth) biocarrier and inoculated in a similar manner to the glass bead reactor. Degradation of CT in this reactor appeared to be suppressed, possibly due to the availability of iron to the bacteria from the biocarrier. While these experiments indicate it should be possible to design a bioreactor utilizing Pseudomonas sp. KC to degrade CT in contaminated water, the suppression of degradative activity by iron will require special precautions to be taken in reactor design.

  4. An additional simple denitrification bioreactor using packed gel envelopes applicable to industrial wastewater treatment.

    PubMed

    Morita, Masahiko; Uemoto, Hiroaki; Watanabe, Atsushi

    2007-08-15

    A simple denitrification bioreactor for nitrate-containing wastewater without organic compounds was developed. This bioreactor consisted of packed gel envelopes in a single tank. Each envelope comprised two plates of gels containing Paracoccus denitrificans cells with an internal space between the plates. As an electron donor for denitrification, ethanol was injected into the internal space and not directly into the wastewater. P. denitrificans cells in the gel reduced nitrate to nitrogen gas by using the injected ethanol. Nitrate-containing desulfurization wastewater derived from a coal-fired thermal power plant was continuously treated with 20 packed gel envelopes (size, 1,000 x 900 x 12 mm; surface area, 1.44 m(2)) in a reactor tank (volume 1.5 m(3)). When the total nitrogen concentration in the inflow was around 150 mg-N x L(-1), the envelopes removed approximately 60-80% of the total nitrogen, and the maximum nitrogen removal rate was 5.0 g-N x day(-1) per square meter of the gel surface. This value corresponded to the volumetric nitrogen removal performance of 0.109 kg-N x m(-3) x day(-1). In each envelope, a high utilization efficiency of the electron donor was attained, although more than the double amount of the electron donor was empirically injected in the present activated sludge system to achieve denitrification when compared with the theoretical value. The bioreactor using the envelopes would be extremely effective as an additional denitrification system because these envelopes can be easily installed in the vacant spaces of preinstalled water treatment systems, without requiring additional facilities for removing surplus ethanol and sludge. PMID:17252606

  5. Comparison of bioreactors with different kinds of submerged packed beds for domestic wastewater treatment.

    PubMed

    Nacheva, P Mijaylova; Moeller Chávez, G; Bustos, C; Garzón Zúñiga, M A; Hornelas Orozco, Y

    2008-01-01

    The performance of aerobic submerged packed bed reactors was studied for the treatment of domestic wastewater using different kinds of packing materials with high specific areas (760-1,200 m(2)/m(3)). The tested materials were ceramic spheres, crushed tezontle, grains of high density polyethylene (HDPE), of low density polyethylene (LDPE) and of polypropylene (PP), cubes of polyurethane (PU) and polyethylene tape (SESSIL). The bioreactors were operated in continuous regime, applying organic loads in the range of 0.8-6.0 g COD.m(-2).d(-1). The obtained specific COD removal rates were very similar in all the reactors when they were operated at organic loads up to 2.0 g COD.m(-2).d(-1), after which differences in effectiveness appeared and the best results were determined in the reactors with SESSIL, LDPE and PU. Very low TSS, O&G and turbidity were obtained in all the effluents. The NH(3)-N and TN removals were dependent on the dissolved oxygen (DO) concentration and the removals at DO of 5 mg/l were 84-99% and 61-74% respectively. The best removals were determined in the reactors with PU, SESSIL and LDPE. The reactor with tezontle had also a good performance when operated with loads up to 1.0 g TN.m(-2).d(-1). The best phosphate removals (38-49%) were obtained in the reactors with PU, tezontle, ceramic sheres and SESSIL.

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

    PubMed

    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. PMID:24089597

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

    PubMed Central

    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 (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. PMID:24089597

  8. Perchlorate remediation using packed-bed bioreactors and electricity generation in microbial fuel cells (MFCs)

    NASA Astrophysics Data System (ADS)

    Min, Booki

    Two pilot-scale fixed bed bioreactors were operated in continuous mode in order to treat groundwater contaminated by perchlorate. The bioreactors were constructed and operated side-by-side at the Texas Street Well Facility in Redlands, California. Each reactor was packed with either sand or plastic media. A perchlorate-reducing bacterium, Dechlorosoma sp. KJ, was used to inoculate the bioreactors. Perchlorate was successfully removed down to a non-detectable level (<4mug/L) in both bioreactors with acetate as a carbon source and nutrients at loading rates less than 0.063 L/s (1 gpm; 0.34 L/m2s). The sand medium bioreactor could achieve complete-perchlorate removal up to flow rate of 0.126 L/s. A regular backwashing cycle (once a week) was an important factor for completely removing perchlorate in groundwater. Power generation directly from pure or mixed organic matter was examined using microbial fuel cells (MFCs), which were run either in batch or continuous mode. In batch experiments, both a pure culture (Geobactor metallireducens) and a mixed culture (wastewater inoculum) were used as the biocatalyst, and acetate was added as substrate in the anode chamber of the MFC. Power output in a membrane MFC with either inoculum was essentially the same, with 40 +/- 1 mW/m2 for G. metallireducens and 38 +/- 1 mW/m2 for mixed culture. A different type of the MFC containing a salt bridge instead of a membrane system was examined to generate power using the same substrate and pure culture as used in the membrane MFC. Power output in the salt bridge MFC was 2.2 mW/m 2. It was found that the lower power output was directly attributed to the higher internal resistance of the salt bridge system (19920 +/- 50 O) in comparison with that of the membrane system (1286 +/- 1 O). Continuous electricity generation was examined in a flat plate microbial fuel cell (FPMFC) using domestic wastewater and specific organic substrates. The FPMFC, containing a combined electrode/proton exchange

  9. Gaseous hexane biodegradation by Fusarium solani in two liquid phase packed-bed and stirred-tank bioreactors.

    PubMed

    Arriaga, Sonia; Muñoz, Raúl; Hernández, Sergio; Guieysse, Benoit; Revah, Sergio

    2006-04-01

    Biofiltration of hydrophobic volatile pollutants is intrinsically limited by poor transfer of the pollutants from the gaseous to the liquid biotic phase, where biodegradation occurs. This study was conducted to evaluate the potential of silicone oil for enhancing the transport and subsequent biodegradation of hexane by the fungus Fusarium solani in various bioreactor configurations. Silicone oil was first selected among various solvents for its biocompatibility, nonbiodegradability, and good partitioning properties toward hexane. In batch tests, the use of silicone oil improved hexane specific biodegradation by approximately 60%. Subsequent biodegradation experiments were conducted in stirred-tank (1.5 L) and packed-bed (2.5 L) bioreactors fed with a constant gaseous hexane load of 180 g x m(-3)(reactor) x h(-1) and operated for 12 and 40 days, respectively. In the stirred reactors, the maximum hexane elimination capacity (EC) increased from 50 g x m(-3)(reactor) x h(-1) (removal efficiency, RE of 28%) in the control not supplied with silicone oil to 120 g x m(-3)(reactor) x h(-1) in the biphasic system (67% RE). In the packed-bed bioreactors, the maximum EC ranged from 110 (50% RE) to 180 g x m(-3)(reactor) x h(-1) (> 90% RE) in the control and two-liquid-phase systems, respectively. These results represent, to the best of our knowledge, the first reported case of fungi use in a two-liquid-phase bioreactor and the highest hexane removal capacities so far reported in biofilters. PMID:16646479

  10. Gaseous hexane biodegradation by Fusarium solani in two liquid phase packed-bed and stirred-tank bioreactors.

    PubMed

    Arriaga, Sonia; Muñoz, Raúl; Hernández, Sergio; Guieysse, Benoit; Revah, Sergio

    2006-04-01

    Biofiltration of hydrophobic volatile pollutants is intrinsically limited by poor transfer of the pollutants from the gaseous to the liquid biotic phase, where biodegradation occurs. This study was conducted to evaluate the potential of silicone oil for enhancing the transport and subsequent biodegradation of hexane by the fungus Fusarium solani in various bioreactor configurations. Silicone oil was first selected among various solvents for its biocompatibility, nonbiodegradability, and good partitioning properties toward hexane. In batch tests, the use of silicone oil improved hexane specific biodegradation by approximately 60%. Subsequent biodegradation experiments were conducted in stirred-tank (1.5 L) and packed-bed (2.5 L) bioreactors fed with a constant gaseous hexane load of 180 g x m(-3)(reactor) x h(-1) and operated for 12 and 40 days, respectively. In the stirred reactors, the maximum hexane elimination capacity (EC) increased from 50 g x m(-3)(reactor) x h(-1) (removal efficiency, RE of 28%) in the control not supplied with silicone oil to 120 g x m(-3)(reactor) x h(-1) in the biphasic system (67% RE). In the packed-bed bioreactors, the maximum EC ranged from 110 (50% RE) to 180 g x m(-3)(reactor) x h(-1) (> 90% RE) in the control and two-liquid-phase systems, respectively. These results represent, to the best of our knowledge, the first reported case of fungi use in a two-liquid-phase bioreactor and the highest hexane removal capacities so far reported in biofilters.

  11. Lipase production by solid-state fermentation: cultivation conditions and operation of tray and packed-bed bioreactors.

    PubMed

    Gutarra, Melissa L E; Cavalcanti, Elisa D C; Castilho, Leda R; Freire, Denise M G; Sant'Anna, Geraldo L

    2005-01-01

    The production of lipase by Penicillium simplicissimum in solid-state fermentation was studied using babassu cake as the basal medium. Tray-type and packed-bed bioreactors were employed. In the former, the influence of temperature; content of the medium, and medium supplementation with olive oil, sugarcane molasses, corn steep liquor, and yeast hydrolysate was studied. For all combinations of supplements, a temperature of 30 degrees C, a moisture content of 70%, and a concentration of carbon source of 6.25% (m/m, dry basis) provided optimum conditions for lipase production. When used as single supplements olive oil and molasses also were able to provide high lipase activities (20 U/g). Using packed-bed bioreactors and molasses-supplemented medium, optimum conditions for enzyme production were air superficial velocities above 55 cm/min and temperatures below 28 degrees C. The lower temperature optimum found for these reactors is probably related to radial heat gradient formation inside the packed bed. Maximum lipase activities obtained in these bioreactors (26.4 U/g) were 30% higher than in tray-type reactors.

  12. Combination of photoreactor and packed bed bioreactor for the removal of ethyl violet from wastewater.

    PubMed

    Chen, Chih-Yu; Yen, Shao-Hsiung; Chung, Ying-Chien

    2014-12-01

    An efficient treatment system that combines a photoreactor and packed bed bioreactor (PBR) was developed and evaluated for treating ethyl violet (EV)-containing wastewater. Initial experiments demonstrated that the optimal operating parameters for the photoreactor in treating EV-containing wastewater were 2h reaction time, pH of 7, and 2 min liquid retention time. Under these conditions, the photocatalytic reaction achieved a 61% EV removal efficiency and resulted in a significant BOD/COD increase in the solution. The results displayed by the coupled photobiological system achieved a removal efficiency of 85% and EC50 of the solution increased by 19 times in a semi-continuous mode when the EV concentration was <150 mg +L(-)(1). The effect of shock loading on the EV removal was temporary but coexisting substrate (glucose and crystal violet) at specific levels would affect the EV removal efficiency of the PBR. Phylogenetic analysis in the PBR indicated that the major bacteria species were Bdellovibrio bacteriovorus, Ralstonia pickettii, Stenotrophomonas maltophilia, and Comamonas sp. Furthermore, the possible degrading mechanisms of this coupled system were demethylation, deethylation, aromatic ring opening, nitrification, and carbon oxidation. The intermediates were characterized using gas chromatography-mass spectrometry analysis. These results indicated that the coupled photobiological system provides an effective method of EV removal. PMID:25259784

  13. Bioaugmented membrane bioreactor (MBR) with a GAC-packed zone for high rate textile wastewater treatment.

    PubMed

    Hai, Faisal Ibney; Yamamoto, Kazuo; Nakajima, Fumiyuki; Fukushi, Kensuke

    2011-03-01

    The long-term performance of a bioaugmented membrane bioreactor (MBR) containing a GAC-packed anaerobic zone for treatment of textile wastewater containing structurally different azo dyes was observed. A unique feeding strategy, consistent with the mode of evolution of separate waste streams in textile plants, was adopted to make the best use of the GAC-zone for dye removal. Dye was introduced through the GAC-zone while the rest of the colorless media was simultaneously fed through the aerobic zone. Preliminary experiments confirmed the importance of coupling the GAC-amended anaerobic zone to the aerobic MBR and also evidenced the efficacy of the adopted feeding strategy. Following this, the robustness of the process under gradually increasing dye-loading was tested. The respective average dye concentrations (mg/L) in the sample from GAC-zone and the membrane-permeate under dye-loadings of 0.1 and 1 g/L.d were as follows: GAC-zone (3, 105), permeate (0, 5). TOC concentration in membrane-permeate for the aforementioned loadings were 3 and 54 mg/L, respectively. Stable decoloration along with significant TOC removal during a period of over 7 months under extremely high dye-loadings demonstrated the superiority of the proposed hybrid process.

  14. Green and efficient production of octyl hydroxyphenylpropionate using an ultrasound-assisted packed-bed bioreactor.

    PubMed

    Lee, Chih Chen; Chen, Hsiao Ching; Ju, Hen Yi; Chen, Jiann Hwa; Kuo, Chia Hung; Chung, Yi Lin; Liu, Yung Chuan; Shieh, Chwen Jen

    2012-04-01

    A solvent-free system to produce octyl hydroxyphenylpropionate (OHPP) from p-hydroxyphenylpropionic acid (HPPA) and octanol using immobilized lipase (Novozym® 435) as a catalyst in an ultrasound-assisted packed-bed bioreactor was investigated. Response-surface methodology (RSM) and a three-level-three-factor Box-Behnken design were employed to evaluate the effects of reaction temperature (x₁), flow rate (x₂) and ultrasonic power (x₃) on the percentage of molar production of OHPP. The results indicate that the reaction temperature and flow rate were the most important variables in optimizing the production of OHPP. Based on a ridge max analysis, the optimum conditions for OHPP synthesis were predicted to consist of a reaction temperature of 65°C, a flow rate of 0.05 ml/min and an ultrasonic power of 1.74 W/cm² with a yield of 99.25%. A reaction was performed under these optimal conditions, and a yield of 99.33 ± 0.1% was obtained. PMID:22120649

  15. Bioreactor

    NASA Technical Reports Server (NTRS)

    1996-01-01

    The NASA Bioreactor provides a low turbulence culture environment which promotes the formation of large, three-dimensional cell clusters. Due to their high level of cellular organization and specialization, samples constructed in the bioreactor more closely resemble the original tumor or tissue found in the body. NASA-sponsored bioreactor research has been instrumental in helping scientists to better understand normal and cancerous tissue development. In cooperation with the medical community, the bioreactor design is being used to prepare better models of human colon, prostate, breast and ovarian tumors. Cartilage, bone marrow, heart muscle, skeletal muscle, pancreatic islet cells, liver and kidney are just a few of the normal tissues currently being cultured in rotating bioreactors by investigators

  16. Nitrogen and Phosphorus Removal from Wastewater Treatment Plant Effluent via Bacterial Sulfate Reduction in an Anoxic Bioreactor Packed with Wood and Iron

    PubMed Central

    Yamashita, Takahiro; Yamamoto-Ikemoto, Ryoko

    2014-01-01

    We investigated the removal of nitrogen and phosphate from the effluent of a sewage treatment plant over a long-term operation in bioreactors packed with different combinations of wood and iron, with a trickling filter packed with foam ceramics for nitrification. The average nitrification rate in the trickling filter was 0.17 kg N/m3∙day and remained at 0.11 kg N/m3∙day even when the water temperature was below 15 °C. The denitrification and phosphate removal rates in the bioreactor packed with aspen wood and iron were higher than those in the bioreactor packed with cedar chips and iron. The bioreactor packed with aspen wood and iron continued to remove nitrate and phosphate for >1200 days of operation. The nitrate removal activity of a biofilm attached to the aspen wood from the bioreactor after 784 days of operation was 0.42 g NO3-N/kg dry weight wood∙ day. There was no increase in the amount of dissolved organic matter in the outflow from the bioreactors. PMID:25247426

  17. Nitrogen and phosphorus removal from wastewater treatment plant effluent via bacterial sulfate reduction in an anoxic bioreactor packed with wood and iron.

    PubMed

    Yamashita, Takahiro; Yamamoto-Ikemoto, Ryoko

    2014-09-22

    We investigated the removal of nitrogen and phosphate from the effluent of a sewage treatment plant over a long-term operation in bioreactors packed with different combinations of wood and iron, with a trickling filter packed with foam ceramics for nitrification. The average nitrification rate in the trickling filter was 0.17 kg N/m3∙day and remained at 0.11 kg N/m3∙day even when the water temperature was below 15 °C. The denitrification and phosphate removal rates in the bioreactor packed with aspen wood and iron were higher than those in the bioreactor packed with cedar chips and iron. The bioreactor packed with aspen wood and iron continued to remove nitrate and phosphate for >1200 days of operation. The nitrate removal activity of a biofilm attached to the aspen wood from the bioreactor after 784 days of operation was 0.42 g NO3-N/kg dry weight wood∙ day. There was no increase in the amount of dissolved organic matter in the outflow from the bioreactors.

  18. Biological nitrogen removal from plating wastewater by submerged membrane bioreactor packed with granular sulfur.

    PubMed

    Moon, Jinyoung; Hwang, Yongwoo; Kim, Junbeum; Kwak, Inho

    2016-01-01

    Recent toughened water quality standards have necessitated improvements for existing sewer treatment facilities through advanced treatment processes. Therefore, an advanced treatment process that can be installed through simple modification of existing sewer treatment facilities needs to be developed. In this study, a new submerged membrane bioreactor process packed with granular sulfur (MBR-GS) was developed and operated to determine the biological nitrogen removal behaviors of plating wastewater containing a high concentration of NO3(-). Continuous denitrification was carried out at various nitrogen loading rates at 20 °C using synthetic wastewater, which was comprised of NO3(-) and HCO3(-), and actual plating wastewater, which was collected from the effluent water of a plating company called 'H Metals'. High-rate denitrification in synthetic plating wastewater was accomplished at 0.8 kg NO3(-)-N/m(3)·day at a nitrogen loading rate of 0.9 kg NO3(-)-N/m(3)·day. The denitrification rate further increased in actual plating wastewater to 0.91 kg NO3(-)-N/m(3)·day at a nitrogen loading rate of 1.11 kg NO3(-)-N/m(3)·day. Continuous filtration was maintained for up to 30 days without chemical cleaning with a transmembrane pressure in the range of 20 cmHg. Based on stoichiometry, SO4(2-) production and alkalinity consumption could be calculated theoretically. Experimental alkalinity consumption was lower than the theoretical value. This newly proposed MBR-GS process, capable of high-rate nitrogen removal by compulsive flux, is expected to be applicable as an alternative renovation technique for nitrogen treatment of plating wastewater as well as municipal wastewater with a low C/N ratio. PMID:27533855

  19. Biological nitrogen removal from plating wastewater by submerged membrane bioreactor packed with granular sulfur.

    PubMed

    Moon, Jinyoung; Hwang, Yongwoo; Kim, Junbeum; Kwak, Inho

    2016-01-01

    Recent toughened water quality standards have necessitated improvements for existing sewer treatment facilities through advanced treatment processes. Therefore, an advanced treatment process that can be installed through simple modification of existing sewer treatment facilities needs to be developed. In this study, a new submerged membrane bioreactor process packed with granular sulfur (MBR-GS) was developed and operated to determine the biological nitrogen removal behaviors of plating wastewater containing a high concentration of NO3(-). Continuous denitrification was carried out at various nitrogen loading rates at 20 °C using synthetic wastewater, which was comprised of NO3(-) and HCO3(-), and actual plating wastewater, which was collected from the effluent water of a plating company called 'H Metals'. High-rate denitrification in synthetic plating wastewater was accomplished at 0.8 kg NO3(-)-N/m(3)·day at a nitrogen loading rate of 0.9 kg NO3(-)-N/m(3)·day. The denitrification rate further increased in actual plating wastewater to 0.91 kg NO3(-)-N/m(3)·day at a nitrogen loading rate of 1.11 kg NO3(-)-N/m(3)·day. Continuous filtration was maintained for up to 30 days without chemical cleaning with a transmembrane pressure in the range of 20 cmHg. Based on stoichiometry, SO4(2-) production and alkalinity consumption could be calculated theoretically. Experimental alkalinity consumption was lower than the theoretical value. This newly proposed MBR-GS process, capable of high-rate nitrogen removal by compulsive flux, is expected to be applicable as an alternative renovation technique for nitrogen treatment of plating wastewater as well as municipal wastewater with a low C/N ratio.

  20. Microbial sulfate reduction under sequentially acidic conditions in an upflow anaerobic packed bed bioreactor.

    PubMed

    Jong, Tony; Parry, David L

    2006-07-01

    The aim of this study was to operate an upflow anaerobic packed bed reactor (UAPB) containing sulfate reducing bacteria (SRB) under acidic conditions similar to those found in acid mine drainage (AMD). The UAPB was filled with sand and operated under continuous flow at progressively lower pH and was shown to be capable of supporting sulfate reduction at pH values of 6.0, 5.0, 4.5, 4.0 and 3.5 in a synthetic medium containing 53.5 mmol l(-1) lactate. Sulfate reduction rates of 553-1,052 mmol m(-3) d(-1) were obtained when the influent solution pH was progressively lowered from pH 6.0 to 4.0, under an optimal flow rate of 2.61 ml min(-1). When the influent pH was further lowered to pH 3.5, sulfate reduction was substantially reduced with only about 1% sulfate removed at a rate of 3.35 mmol m(-3) d(-1) after 20 days of operation. However, viable SRB were recovered from the column, indicating that the SRB population was capable of surviving and metabolizing at low levels even at pH 3.5 conditions for at least 20 days. The changes in conductivity in the SRB column did not always occur with changes in pH and redox potential, suggesting that conductivity measurements may be more sensitive to SRB activity and could be used as an additional tool for monitoring SRB activity. The bioreactor containing SRB was able to reduce sulfate and generate alkalinity even when challenged with influent as low as pH 3.5, indicating that such treatment systems have potential for bioremediating highly acidic, sulfate contaminated waste waters.

  1. Comparison of methanogenic community structure and anaerobic process performance treating swine wastewater between pilot and optimized lab scale bioreactors.

    PubMed

    Kim, Woong; Cho, Kyungjin; Lee, Seungyong; Hwang, Seokhwan

    2013-10-01

    To investigate methanogenic community structure and process performance of anaerobic digestion treating swine wastewater at different scale, a pilot plant with 20 m(3) of effective working volume and lab scale methanogenic digester with 6L working volume were operated for 71 days and 6 turnover periods, respectively. During the steady state of anaerobic digestion, COD and VS removal efficiency in pilot plant were 65.3±3.2, 51.6±4.3%, respectively, which was similar to those in lab scale. However, calculated VFAs removal efficiency and methane yield were lower in pilot plant than in lab scale digester. Also, organics removal efficiencies, which consist of total carbohydrates, proteins, and lipids, were different between pilot and lab scale. These results were thought to be due to the ratio of carbohydrates to proteins in the raw swine wastewater. As a result of qualitative microbial analysis, Methanoculleus receptaculii, and Methanoculleus bourgensis, were commonly concerned with methane production.

  2. Continuous production of monoclonal antibody in a packed-bed bioreactor.

    PubMed

    Golmakany, Naghmeh; Rasaee, Mohammad Javad; Furouzandeh, Mehdi; Shojaosadati, Seyed Abbas; Kashanian, Soheila; Omidfar, Kobra

    2005-06-01

    In the present study the growth and MAb (monoclonal antibody) production of a mouse x mouse hybridoma cell producing anti-digoxin MAb was evaluated. The hybridoma cells entrapped within the support matrix Fibra-Cel were cultured in batch and continuous mode following special protocols. Cell-culture studies were performed in a 1-litre spinner basket containing 3 g.litre-1 support matrix. Batch culture was operated with the cell density of 42x10(6) cells. During the 7 days of culture, the medium was sampled daily in order to assess glucose and MAb concentrations and the lactate dehydrogenase released into the culture medium. After a culture period of 72 h, the cell density and MAb concentration were found to be 10.4x10(7) cells/3 g of NWPF (non-woven polyester fibre) discs and 250 microg/ml respectively. This yield gradually decreased to 0.55x10(6) cells/3 g of packaging material and 60 microg/ml respectively at the end of the batch culture. In the continuous-culture studies, the batch culture was initially operated for 64.5 h and then continuous flow was started at the dilution rates of 0.15, 0.2, 0.25 and 0.3 day-1 and finally stabilized at 0.25 day-1 within 288 h (12 days). The MAb concentration at steady state was found to be 116-120 microg/day per ml, and the yield of operation was 62.5 mg/day per ml, which was 3.5 times higher than that of batch culture. In conclusion, a packed-bed bioreactor with the support matrix Fibra-Cel, operated in continuous-feeding mode, is more efficient for large-scale MAb production than a batch culture. On the other hand, by using a continuous-culture system, a better supply of nutrients and removal of inhibitory metabolites and proteolytic enzymes was obtained.

  3. Methanospirillum stamsii sp. nov., a psychrotolerant, hydrogenotrophic, methanogenic archaeon isolated from an anaerobic expanded granular sludge bed bioreactor operated at low temperature.

    PubMed

    Parshina, Sofiya N; Ermakova, Anna V; Bomberg, Malin; Detkova, Ekaterina N

    2014-01-01

    A psychrotolerant hydrogenotrophic methanogen, strain Pt1, was isolated from a syntrophic propionate-oxidizing methanogenic consortium obtained from granulated biomass of a two-stage low-temperature (3-8 °C) anaerobic expanded granular sludge bed (EGSB) bioreactor, fed with a mixture of volatile fatty acids (VFAs) (acetate, propionate and butyrate). The strain was strictly anaerobic, and cells were curved rods, 0.4-0.5×7.5-25 µm, that sometimes formed wavy filaments from 25 to several hundred micrometres in length. Cells stained Gram-negative and were non-sporulating. They were gently motile by means of tufted flagella. The strain grew at 5-37 °C (optimum at 20-30 °C), at pH 6.0-10 (optimum 7.0-7.5) and with 0-0.3 M NaCl (optimum 0 M NaCl). Growth and methane production was found with H2/CO2 and very weak growth with formate. Acetate and yeast extract stimulated growth, but were not essential. The G+C content of the DNA of strain Pt1 was 40 mol%. Phylogenetic analysis based on 16S rRNA gene sequences revealed that strain Pt1 was a member of the genus Methanospirillum and showed 97.5 % sequence similarity to Methanospirillum hungatei JF1(T) and 94 % sequence similarity to Methanospirillum lacunae Ki8-1(T). DNA-DNA hybridization of strain Pt1 with Methanospirillum hungatei JF1(T) revealed 39 % relatedness. On the basis of its phenotypic characteristics and phylogenetic position, strain Pt1 is a representative of a novel species of the genus Methanospirillum, for which the name Methanospirillum stamsii sp. nov. is proposed. The type strain is Pt1(T) ( = DSM 26304(T) = VKM B-2808(T)). PMID:24048867

  4. Energy conservation and production in a packed-bed anaerobic bioreactor

    SciTech Connect

    Pit, W.W. Jr.; Genung, R.K.

    1980-01-01

    Oak Ridge National Laboratory (ORNL) is developing an energy-conserving/ producing wastewater treatment system based on a fixed-film anaerobic bioreactor. The treatment process is based on passing wastewaters upward through the bioreactor for continuous treatment by gravitational settling, biophysical filtration and biological decomposition. A two-year pilot-plant project using a bioreactor designed to treat 5000 gpd has been conducted using raw wastewater on a municipal site in Oak Ridge, Tennessee. Data obtained for the performance of the bioreactor during this project have been analyzed by ORNL and Associated Water and Air Resources Engineers (AWARE), Inc. of Nashville, Tennessee. From these analyses it was estimated that hydraulic loading rates of 0.25 gpm/ft/sup 2/ and hydraulic residence times of 10 hours could be used in designing such bioreactors for the secondary treatment of municipal wastewaters. Conceptual designs for total treatment systems processing up to one million gallons of wastewater per day were developed based on the performance of the pilot plant bioreactor. These systems were compared to activated sludge treatment systems also operating under secondary treatment requirements and were found to consume as little as 30% of the energy required by the activated sludge systems. Economic advantages of the process result from the elimination of operating energy requirements associated with the aeration of aerobic-based processes and with the significant decrease of sludge-handling costs required with conventional activated sludge treatment systems.Furthermore, methane produced by anaerobic fermentation processes occurring during the biological decomposition of carbonaceous wastes also represented a significant and recoverable energy production. For dilute municipal wastewaters this would completely offset the remaining energy required for treatment, while for concentrated industrial wastewater would result in a net production of energy.

  5. Identification, Detection, and Spatial Resolution of Clostridium Populations Responsible for Cellulose Degradation in a Methanogenic Landfill Leachate Bioreactor

    PubMed Central

    Burrell, P. C.; O'Sullivan, C.; Song, H.; Clarke, W. P.; Blackall, L. L.

    2004-01-01

    An anaerobic landfill leachate bioreactor was operated with crystalline cellulose and sterile landfill leachate until a steady state was reached. Cellulose hydrolysis, acidogenesis, and methanogenesis were measured. Microorganisms attached to the cellulose surfaces were hypothesized to be the cellulose hydrolyzers. 16S rRNA gene clone libraries were prepared from this attached fraction and also from the mixed fraction (biomass associated with cellulose particles and in the planktonic phase). Both clone libraries were dominated by Firmicutes phylum sequences (100% of the attached library and 90% of the mixed library), and the majority fell into one of five lineages of the clostridia. Clone group 1 (most closely related to Clostridium stercorarium), clone group 2 (most closely related to Clostridium thermocellum), and clone group 5 (most closely related to Bacteroides cellulosolvens) comprised sequences in Clostridium group III. Clone group 3 sequences were in Clostridium group XIVa (most closely related to Clostridium sp. strain XB90). Clone group 4 sequences were affiliated with a deeply branching clostridial lineage peripherally associated with Clostridium group VI. This monophyletic group comprises a new Clostridium cluster, designated cluster VIa. Specific fluorescence in situ hybridization (FISH) probes for the five groups were designed and synthesized, and it was demonstrated in FISH experiments that bacteria targeted by the probes for clone groups 1, 2, 4, and 5 were very abundant on the surfaces of the cellulose particles and likely the key cellulolytic microorganisms in the landfill bioreactor. The FISH probe for clone group 3 targeted cells in the planktonic phase, and these organisms were hypothesized to be glucose fermenters. PMID:15066839

  6. Kinetics studies of p-cresol biodegradation by using Pseudomonas putida in batch reactor and in continuous bioreactor packed with calcium alginate beads.

    PubMed

    Mathur, A K; Bala, Shashi; Majumder, C B; Sarkar, S

    2010-01-01

    Present study deals with the biodegradation of p-cresol by using Pseudomonas putida in a batch reactor and a continuous bioreactor packed with calcium alginate beads. The maximum specific growth rate of 0.8121 h(-1) was obtained at 200 mg L(-1) concentration of p-cresol in batch reactor. The maximum p-cresol degradation rate was obtained 6.598 mg L(-1) h(-1) at S(o)=200 mg L(-1) and 62.8 mg L(-1) h(-1) at S(o)=500 mg L(-1) for batch reactor and a continuous bioreactor, respectively. The p-cresol degradation rate of continuous bioreactor was 9 to 10-fold higher than those of the batch reactor. It shows that the continuous bioreactor could tolerate a higher concentration of p-cresol. A Haldane model was also used for p-cresol inhibition in batch reactor and a modified equation similar to Haldane model for continuous bioreactor. The Haldane parameters were obtained as µ(max) 0.3398 h(-1), K(s) 110.9574 mg L(-1), and K(I) 497.6169 mg L(-1) in batch reactor. The parameters used in continuous bioreactor were obtained as D(max) 91.801 mg L(-1) h(-1), K(s) 131.292 mg L(-1), and K(I) 1217.7 mg L(-1). The value K(I) of continuous bioreactor is approximately 2.5 times higher than the batch reactor. Higher K(I) value of continuous bioreactor indicates P. putida can grow at high range of p-cresol concentration. The ability of tolerance of higher p-cresol concentrations may be one reason for biofilm attachment on the packed bed in the continuous operation.

  7. Conidia production by Beauveria bassiana (for the biocontrol of a diamondback moth) during solid-state fermentation in a packed-bed bioreactor.

    PubMed

    Kang, S W; Lee, S H; Yoon, C S; Kim, S W

    2005-01-01

    Conidia of Beauveria bassiana CS-1, which have the potential for the control of the diamondback moth (Plutella xylostella), were produced by solid-state fermentation (SSF) using a packed-bed bioreactor with rice straw and wheat bran. As the packing density and the bed height were increased, the production of conidia decreased. In a packed-bed bioreactor under no aeration and no addition of polypropylene (PP) foam (control), the total average of conidia was 4.9 x 10(8) g-1. The production of conidia was affected more by the addition of PP foam as an inert support than forced aeration and was approx. 23 times higher than that of the control. The total average of conidia produced by B. bassiana was 1.1-1.2 x 10(10) g-1 .

  8. Anaerobium acetethylicum gen. nov., sp. nov., a strictly anaerobic, gluconate-fermenting bacterium isolated from a methanogenic bioreactor.

    PubMed

    Patil, Yogita; Junghare, Madan; Pester, Michael; Müller, Nicolai; Schink, Bernhard

    2015-10-01

    A novel strictly anaerobic, mesophilic bacterium was enriched and isolated with gluconate as sole substrate from a methanogenic sludge collected from a biogas reactor. Cells of strain GluBS11T stained Gram-positive and were non-motile, straight rods, measuring 3.0-4.5 × 0.8-1.2 μm. The temperature range for growth was 15-37 °C, with optimal growth at 30 °C, the pH range was 6.5-8.5, with optimal growth at pH 7, and the generation time under optimal conditions was 60 min. API Rapid 32A reactions were positive for α-galactosidase, α-glucosidase and β-glucosidase and negative for catalase and oxidase. A broad variety of substrates was utilized, including gluconate, glucose, fructose, maltose, sucrose, lactose, galactose, melezitose, melibiose, mannitol, erythritol, glycerol and aesculin. Products of gluconate fermentation were ethanol, acetate, formate, H2 and CO2. Neither sulfate nor nitrate served as an electron acceptor. Predominant cellular fatty acids (>10 %) were C14 : 0, C16 : 0, C16 : 1ω7c/iso-C15 : 0 2-OH and C18 : 1ω7c. The DNA G+C content of strain GluBS11T was 44.1 mol%. Phylogenetic analysis based on 16S rRNA gene sequence data revealed that strain GluBS11T is a member of subcluster XIVa within the order Clostridiales. The closest cultured relatives are Clostridium herbivorans (93.1 % similarity to the type strain), Clostridium populeti (93.3 %), Eubacterium uniforme (92.4 %) and Clostridium polysaccharolyticum (91.5 %). Based on this 16S rRNA gene sequence divergence (>6.5 %) as well as on chemotaxonomic and phenotypic differences from these taxa, strain GluBS11T is considered to represent a novel genus and species, for which the name Anaerobium acetethylicum gen. nov., sp. nov. is proposed. The type strain of Anaerobium acetethylicum is GluBS11T ( = LMG 28619T = KCTC 15450T = DSM 29698T). PMID:26297346

  9. Removal of nitrate and hexavalent uranium from groundwater by sequential treatment in bioreactors packed with elemental sulfur and zero-valent iron.

    PubMed

    Luna-Velasco, Antonia; Sierra-Alvarez, Reyes; Castro, Beatriz; Field, Jim A

    2010-12-15

    The bioreduction of soluble hexavalent uranium (U(VI)) to insoluble tetravalent uranium (U(IV)) is an attractive bioremediation strategy for the clean-up of contaminated groundwater. High levels of the common occurring co-contaminant, nitrate (NO3(-)), can potentially interfere with uranium bioremediation. In this study, treatment of a synthetic groundwater containing a mixture of NO3(-) and U(VI) was investigated in a sulfur-limestone autotrophic denitrifying (SLAD) bioreactor that was coupled in series with a bioreactor packed with zero-valent iron (Fe(0), ZVI) and sand. An additional aim of the study was to explore the possible role of biological activity in enhancing the reduction of U(VI) by Fe(0). The SLAD reactor removed NO3(-) efficiently (99.8%) at loadings of up to 20 mmol NO3(-) L(r)(-1) d(-1), with near stoichiometric conversion to benign dinitrogen gas (N(2)). The ZVI bioreactor subsequently removed uranium (99.8%) at high (0.22 mM) and low (0.02 mM) influent concentrations of the radionuclide. Aqueous uranium was reliably eliminated to below the maximum contaminant level of 30 µg L(-1) (0.13 µM) when the ZVI reactor was operated at average empty bed hydraulic retention times as low as 2.3 h, demonstrating the feasibility of the sequential treatment strategy in packed bed bioreactors. Sequential extraction of the ZVI reactor packing confirmed that uranium was immobilized as U(IV). Uranium removal was enhanced by microbial activity as confirmed by the increased rate of uranium removal in batch assays inoculated with effluent from the ZVI bioreactor and spiked with Fe(0) compared to abiotic controls.

  10. Influence of operational parameters on the fluid-side mass transfer resistance observed in a packed bed bioreactor.

    PubMed

    Hussain, Amir; Kangwa, Martin; Abo-Elwafa, Ahmed Gad; Fernandez-Lahore, Marcelo

    2015-01-01

    The influence of mass transfer on productivity as well as the performance of packed bed bioreactor was determined by varying a number of parameters; flow rate, glucose concentration and polymers (chitosan). Saccharomyces cerevisiae cells were immobilized in chitosan and non-chitosan coated alginate beads to demonstrate the effect on external mass transfer by substrate consumption time, lag phase and ethanol production. The results indicate that coating has a significant effect on the lag phase duration, being 30-40 min higher than non-coated beads. After lag phase, no significant change was observed in both types of beads on consumption of glucose with the same flow rate. It was observed that by increasing flow rates; lag phase and glucose consumption time decreased. The reason is due to the reduction of external mass transfer as a result of increase in flow rate as glucose is easily transported to and from the beads surface by diffusion. It is observed that chitosan acts as barrier for transfer of substrate and products, in and out of beads, at initial time of fermentation as it shows longer lag phase for chitosan coated beads than non-coated. Glucose consumption at low flow rate was lower as compared to higher flow rates. The optimum combination of parameters consisting of higher flow rates 30-90 ml/min and between 10 and 20 g/l of glucose was found for maximum production of ethanol.

  11. Operational parameters and their influence on particle-side mass transfer resistance in a packed bed bioreactor.

    PubMed

    Hussain, Amir; Kangwa, Martin; Yumnam, Nivedita; Fernandez-Lahore, Marcelo

    2015-12-01

    The influence of internal mass transfer on productivity as well as the performance of packed bed bioreactor was determined by varying a number of parameters; chitosan coating, flow rate, glucose concentration and particle size. Saccharomyces cerevisiae cells were immobilized in chitosan and non-chitosan coated alginate beads to demonstrate the effect on particle side mass transfer on substrate consumption time, lag phase and ethanol production. The results indicate that chitosan coating, beads size, glucose concentration and flow rate have a significant effect on lag phase duration. The duration of lag phase for different size of beads (0.8, 2 and 4 mm) decreases by increasing flow rate and by decreasing the size of beads. Moreover, longer lag phase were found at higher glucose medium concentration and also with chitosan coated beads. It was observed that by increasing flow rates; lag phase and glucose consumption time decreased. The reason is due to the reduction of external (fluid side) mass transfer as a result of increase in flow rate as glucose is easily transported to the surface of the beads. Varying the size of beads is an additional factor: as it reduces the internal (particle side) mass transfer by reducing the size of beads. The reason behind this is the distance for reactants to reach active site of catalyst (cells) and the thickness of fluid created layer around alginate beads is reduced. The optimum combination of parameters consisting of smaller beads size (0.8 mm), higher flow rate of 90 ml/min and glucose concentration of 10 g/l were found to be the maximum condition for ethanol production.

  12. Process parameters study of α-amylase production in a packed-bed bioreactor under solid-state fermentation with possibility of temperature monitoring.

    PubMed

    Derakhti, Sorour; Shojaosadati, Seyed Abbas; Hashemi, Maryam; Khajeh, Khosro

    2012-01-01

    Production of α-amylase in a laboratory-scale packed-bed bioreactor by Bacillus sp. KR-8104 under solid-state fermentation (SSF) with possibility of temperature control and monitoring was studied using wheat bran (WB) as a solid substrate. The simultaneous effects of aeration rate, initial substrate moisture, and incubation temperature on α-amylase production were evaluated using response surface methodology (RSM) based on a Box-Behnken design. The optimum conditions for attaining the maximum production of α-amylase were 37°C, 72% (w/w) initial substrate moisture, and 0.15 L/min aeration. The average enzyme activity obtained under the optimized conditions was 473.8 U/g dry fermented substrate. In addition, it was observed that the production of enzyme decreased from the bottom of the bioreactor to the top.

  13. Enhancing the Bioconversion of Winery and Olive Mill Waste Mixtures into Lignocellulolytic Enzymes and Animal Feed by Aspergillus uvarum Using a Packed-Bed Bioreactor.

    PubMed

    Salgado, José Manuel; Abrunhosa, Luís; Venâncio, Armando; Domínguez, José Manuel; Belo, Isabel

    2015-10-28

    Wineries and olive oil industries are dominant agro-industrial activities in southern European regions. Olive pomace, exhausted grape marc, and vine shoot trimmings are lignocellulosic residues generated by these industries, which could be valued biotechnologically. In the present work these residues were used as substrate to produce cellulases and xylanases through solid-state fermentation using Aspergillus uvarum MUM 08.01. For that, two factorial designs (3(2)) were first planned to optimize substrate composition, temperature, and initial moisture level. Subsequently, the kinectics of cellulolytic enzyme production, fungal growth, and fermented solid were characterized. Finally, the process was performed in a packed-bed bioreactor. The results showed that cellulase activity improved with the optimization processes, reaching 33.56 U/g, and with the packed-bed bioreactor aeration of 0.2 L/min, reaching 38.51 U/g. The composition of fermented solids indicated their potential use for animal feed because cellulose, hemicellulose, lignin, and phenolic compounds were partially degraded 28.08, 10.78, 13.3, and 28.32%, respectively, crude protein was increased from 8.47 to 17.08%, and the mineral contents meet the requirements of main livestock.

  14. Influence of the addition of sulphate and ferric ions in a methanogenic anaerobic packed-bed reactor treating gasoline-contaminated water.

    PubMed

    Fernandes, B S; Chinalia, F A; Sarti, A; Silva, A J; Foresti, E; Zaiat, M

    2006-01-01

    Benzene, toluene and xylene (BTX) are relatively soluble aromatic compounds of gasoline. Gasoline storage tank leakages generally lead to an extensive contamination of groundwater. In the natural environment for instance, these compounds might be biodegraded under a variety of reducing potentials. The objective of this work was to examine the influence of the addition of sulphate and Fe(OH)3 in a methanogenic horizontal-flow anaerobic immobilized-biomass reactor treating gasoline-contaminated water. Three different conditions were evaluated: methanogenic, sulphidogenic and sulphidogenic with the addition of ferric ions. Methanogenic condition showed the higher BTX degradation rates and the addition of sulphate negatively affected BTX removal rates with the production of H2S. However, the addition of ferric ions resulted in the precipitation of sulphur, improving BTX degradation by the consortium. Metanosphaera sp., Methanosarcina barkeri and Methanosaeta concilii were identified in the consortium by means of 16S and directly related to the addition of ferric ions.

  15. Experimental studies and two-dimensional modelling of a packed bed bioreactor used for production of galacto-oligosaccharides (GOS) from milk whey.

    PubMed

    Sen, Pramita; Bhattacharjee, Chiranjib; Bhattacharya, Pinaki

    2016-03-01

    In the present study, extensive experimental investigations and detailed theoretical analysis of a two-dimensional packed bed bioreactor, employed for the production of galacto-oligosaccharides (GOS) from milk whey were performed. Model equations, in one- and two-dimensions, capable of predicting the substrate concentration distribution in the bioreactor were developed by coupling mass balance equation with appropriate velocity distribution equation and solved numerically. Validation of the proposed model equations was done by a set of experimental data obtained from the bioreactor. The effects of reactor to catalyst particle diameter ratio (d t/d p), feed flowrate (10(-6)-10(-9) m(3) s(-1)), and initial lactose concentration (50-200 kg m(-3)) on substrate concentration distribution were investigated in detail. While, the distribution of substrate concentration in axial direction was independent of d t/d p, it was observed that for d t/d p <40, significant radial concentration distribution existed. It was further observed that the substrate conversion and product yield obtained experimentally showed an excellent agreement (97 ± 2 %) with the results predicted by the two-dimensional model equation, whereas, the results predicted by the one-dimensional model equation did not lie within the desired confidence level (<90 %). The results were confirmed by both curve fitting and statistical analysis. The prediction of substrate concentration distribution in axial and radial directions using the developed two-dimensional model equation is necessary for computing the bioreactor volume to achieve the desired GOS yield.

  16. Experimental studies and two-dimensional modelling of a packed bed bioreactor used for production of galacto-oligosaccharides (GOS) from milk whey.

    PubMed

    Sen, Pramita; Bhattacharjee, Chiranjib; Bhattacharya, Pinaki

    2016-03-01

    In the present study, extensive experimental investigations and detailed theoretical analysis of a two-dimensional packed bed bioreactor, employed for the production of galacto-oligosaccharides (GOS) from milk whey were performed. Model equations, in one- and two-dimensions, capable of predicting the substrate concentration distribution in the bioreactor were developed by coupling mass balance equation with appropriate velocity distribution equation and solved numerically. Validation of the proposed model equations was done by a set of experimental data obtained from the bioreactor. The effects of reactor to catalyst particle diameter ratio (d t/d p), feed flowrate (10(-6)-10(-9) m(3) s(-1)), and initial lactose concentration (50-200 kg m(-3)) on substrate concentration distribution were investigated in detail. While, the distribution of substrate concentration in axial direction was independent of d t/d p, it was observed that for d t/d p <40, significant radial concentration distribution existed. It was further observed that the substrate conversion and product yield obtained experimentally showed an excellent agreement (97 ± 2 %) with the results predicted by the two-dimensional model equation, whereas, the results predicted by the one-dimensional model equation did not lie within the desired confidence level (<90 %). The results were confirmed by both curve fitting and statistical analysis. The prediction of substrate concentration distribution in axial and radial directions using the developed two-dimensional model equation is necessary for computing the bioreactor volume to achieve the desired GOS yield. PMID:26758713

  17. Competition and coexistence of sulfate-reducing bacteria, acetogens and methanogens in a lab-scale anaerobic bioreactor as affected by changing substrate to sulfate ratio.

    PubMed

    Dar, Shabir A; Kleerebezem, Robbert; Stams, Alfons J M; Kuenen, J Gijs; Muyzer, Gerard

    2008-04-01

    The microbial population structure and function of natural anaerobic communities maintained in lab-scale continuously stirred tank reactors at different lactate to sulfate ratios and in the absence of sulfate were analyzed using an integrated approach of molecular techniques and chemical analysis. The population structure, determined by denaturing gradient gel electrophoresis and by the use of oligonucleotide probes, was linked to the functional changes in the reactors. At the influent lactate to sulfate molar ratio of 0.35 mol mol(-1), i.e., electron donor limitation, lactate oxidation was mainly carried out by incompletely oxidizing sulfate-reducing bacteria, which formed 80-85% of the total bacterial population. Desulfomicrobium- and Desulfovibrio-like species were the most abundant sulfate-reducing bacteria. Acetogens and methanogenic Archaea were mostly outcompeted, although less than 2% of an acetogenic population could still be observed at this limiting concentration of lactate. In the near absence of sulfate (i.e., at very high lactate/sulfate ratio), acetogens and methanogenic Archaea were the dominant microbial communities. Acetogenic bacteria represented by Dendrosporobacter quercicolus-like species formed more than 70% of the population, while methanogenic bacteria related to uncultured Archaea comprising about 10-15% of the microbial community. At an influent lactate to sulfate molar ratio of 2 mol mol(-1), i.e., under sulfate-limiting conditions, a different metabolic route was followed by the mixed anaerobic community. Apparently, lactate was fermented to acetate and propionate, while the majority of sulfidogenesis and methanogenesis were dependent on these fermentation products. This was consistent with the presence of significant levels (40-45% of total bacteria) of D. quercicolus-like heteroacetogens and a corresponding increase of propionate-oxidizing Desulfobulbus-like sulfate-reducing bacteria (20% of the total bacteria). Methanogenic Archaea

  18. Bioreactor performance and methanogenic population dynamics in a low-temperature (5-18 °C) anaerobic fixed-bed reactor.

    PubMed

    Zhang, Dongdong; Zhu, Wanbin; Tang, Can; Suo, Yali; Gao, Lijuan; Yuan, Xufeng; Wang, Xiaofen; Cui, Zongjun

    2012-01-01

    The effect of temperature on the functionality of microbial community structure in a low temperature, anaerobic fixed-bed reactor was studied by decreasing the operating temperature from 18 °C to 5 °C. The reactor was productive within 20 days and produced stable methane content in biogas (above 77%) throughout the trial period. At 17 °C and 15 °C, chemical oxygen demand (COD) removal efficiency and biogas production of reactor were significantly reduced. These might be temperature thresholds when fixed-bed reactors are operated under low temperatures. The methanogen community composition was analyzed using 16S rRNA gene clone library screening and quantitative PCR. At low ambient temperatures, Methanomicrobiales were dominant methanogens, and they preferentially adhered to the carbon fiber carrier. The results indicated that 16S rRNA levels of Methanomicrobiales and Methanosaetaceae in adhering sludge were higher than in deposited sludge, and they all contributed to the efficient performance of the fixed-bed reactor at low operating temperatures. PMID:22137750

  19. Simultaneous removal of chromate and nitrate in a packed-bed bioreactor using biodegradable meal box as carbon source and biofilm carriers.

    PubMed

    Li, Jie; Jin, Ruofei; Liu, Guangfei; Tian, Tian; Wang, Jing; Zhou, Jiti

    2016-05-01

    An up-flow packed-bed bioreactor was constructed to investigate the simultaneous removal of chromate and nitrate using biodegradable meal box as carbon source and biofilm carriers. The bioreactor was operated for 164days with varying influent Cr(VI) concentrations (2.0-50.0mg/L) and hydraulic retention times (HRT, 10-24h). It was shown that complete denitrification and Cr(VI) reduction could be achieved when influent Cr(VI) concentrations were lower than 20mg/L with a HRT of 17h. Shortening the HRT could significantly reduce the effluent CODcr. It was also observed that Cr(III) was mainly immobilized on the biofilm. Further investigation on Cr distribution in the biofilm compartments indicated that Cr(VI) reduction occurred in all compartments and the intercellular Cr was dominant. High-throughput sequencing analysis showed that Proteobacteria, Bacteroidetes and Firmicutes were the dominant phyla in the biofilm and Cr(VI) stress had a negative effect on the abundance of most bacteria. PMID:26896715

  20. Tapered bed bioreactor

    DOEpatents

    Scott, Charles D.; Hancher, Charles W.

    1977-01-01

    A vertically oriented conically shaped column is used as a fluidized bed bioreactor wherein biologically catalyzed reactions are conducted in a continuous manner. The column utilizes a packing material a support having attached thereto a biologically active catalytic material.

  1. The influence of hydrolysis induced biopolymers from recycled aerobic sludge on specific methanogenic activity and sludge filterability in an anaerobic membrane bioreactor.

    PubMed

    Buntner, D; Spanjers, H; van Lier, J B

    2014-03-15

    The objective of the present study was to evaluate the impact of excess aerobic sludge on the specific methanogenic activity (SMA), in order to establish the maximum allowable aerobic sludge loading. In batch tests, different ratios of aerobic sludge to anaerobic inoculum were used, i.e. 0.03, 0.05, 0.10 and 0.15, showing that low ratios led to an increased SMA. However, the ratio 0.15 caused more than 20% SMA decrease. In addition to the SMA tests, the potential influence of biopolymers and extracellular substances, that are generated as a result of excess aerobic sludge hydrolysis, on membrane performance was determined by assessing the fouling potential of the liquid broth, taking into account parameters such as specific resistance to filtration (SRF) and supernatant filterability (SF). Addition of aerobic sludge to the anaerobic biomass resulted in a high membrane fouling potential. The increase in biopolymers could be ascribed to aerobic sludge hydrolysis. A clear positive correlation between the concentration of the colloidal fraction of biopolymer clusters (cBPC) and the SRF was observed and a negative correlation between the cBPC and the SF measured at the end of the above described SMA tests. The latter implies that sludge filtration resistance increases when more aerobic sludge is hydrolyzed, and thus more cBPC is released. During AnMBR operation, proteins significantly contributed to sludge filterability decrease expressed as SRF and SF, whereas the carbohydrate fraction of SMP was of less importance due to low concentrations. On the contrary, carbohydrates seemed to improve filterability and diminish SRF of the sludge. Albeit, cBPC increase caused an increase in mean TMP during the AnMBR operation, confirming that cBPC is positively correlated to membrane fouling.

  2. Fluid and mass transport modelling to drive the design of cell-packed hollow fibre bioreactors for tissue engineering applications.

    PubMed

    Shipley, Rebecca J; Waters, Sarah L

    2012-12-01

    A model for fluid and mass transport in a single module of a tissue engineering hollow fibre bioreactor (HFB) is developed. Cells are seeded in alginate throughout the extra-capillary space (ECS), and fluid is pumped through a central lumen to feed the cells and remove waste products. Fluid transport is described using Navier-Stokes or Darcy equations as appropriate; this is overlaid with models of mass transport in the form of advection-diffusion-reaction equations that describe the distribution and uptake/production of nutrients/waste products. The small aspect ratio of a module is exploited and the option of opening an ECS port is explored. By proceeding analytically, operating equations are determined that enable a tissue engineer to prescribe the geometry and operation of the HFB by ensuring the nutrient and waste product concentrations are consistent with a functional cell population. Finally, results for chondrocyte and cardiomyocyte cell populations are presented, typifying two extremes of oxygen uptake rates.

  3. Involvement of ligninolytic enzymes of Phanerochaete chrysosporium in treating the textile effluent containing Astrazon Red FBL in a packed-bed bioreactor.

    PubMed

    Sedighi, M; Karimi, A; Vahabzadeh, F

    2009-09-30

    The effect of Tween80, Mn(II) and veratryl alcohol (VA) on the production of ligninolytic enzymes of Phanerochaete chrysosporium in a packed-bed bioreactor using small pieces of Kissiris as carrier, was investigated. The results of the enzyme activities were noticeable in terms of decolorization and COD removal of the textile effluent containing an azo dye (Astrazon Red FBL). No dilution was made on the tested textile effluent and it was not sterilized, also. Maximum decolorization of the dye (87%) and COD removal (42%), both occurred when only Tween80 (0.05%, w/v) was added to the effluent. The maximum activities of lignin peroxidase (LiP) and manganese peroxidase (MnP) were (U/l): 17 and 52, respectively. The role of MnP was pronounced in the dye decolorization process, while the influence of LiP was noticeable on COD removal. The reusability of the original biomass was examined by replacing undiluted textile effluent (i.e., five times). The cellular performance of the original biomass in repeated-batch operations was promising.

  4. Analysis of bacterial diversity and efficiency of continuous removal of Victoria Blue R from wastewater by using packed-bed bioreactor.

    PubMed

    Chen, Chih-Yu; Wang, Guey-Horng; Tseng, I-Hung; Chung, Ying-Chien

    2016-02-01

    The characteristics of a packed-bed bioreactor (PBB) for continuously removing Victoria Blue R (VBR) from an aqueous solution were determined. The effects of various factors including liquid retention time (RT), VBR concentration, shock loading, and coexisting compounds on the VBR removal and bacterial community in a continuous system were investigated. The intermediates of degraded VBR and the acute toxicity of the effluent from PBB were analyzed. When the VBR concentration was lower than 400 mg/l for a two-day retention time (RT), 100% removal was achieved. During continuous operation, the efficiency initially varied with the VBR concentration and RT, but gradually increased in one to two days. Furthermore, the acute toxicity of the effluent reduced by a factor of 21.25-49.61, indicating that the PBB can be successfully operated under turbulent environmental conditions. VBR degradation involved stepwise demethylation and yielded partially dealkylated VBR species. Phylogenetic analysis showed that the dominant phylum in the PBB was Proteobacteria and that Aeromonas hydrophila dominated during the entire operating period. The characteristics of the identified species showed that the PBB is suitable for processes such as demethylation, aromatic ring opening, carbon oxidation, nitrification, and denitrification.

  5. Analysis of bacterial diversity and efficiency of continuous removal of Victoria Blue R from wastewater by using packed-bed bioreactor.

    PubMed

    Chen, Chih-Yu; Wang, Guey-Horng; Tseng, I-Hung; Chung, Ying-Chien

    2016-02-01

    The characteristics of a packed-bed bioreactor (PBB) for continuously removing Victoria Blue R (VBR) from an aqueous solution were determined. The effects of various factors including liquid retention time (RT), VBR concentration, shock loading, and coexisting compounds on the VBR removal and bacterial community in a continuous system were investigated. The intermediates of degraded VBR and the acute toxicity of the effluent from PBB were analyzed. When the VBR concentration was lower than 400 mg/l for a two-day retention time (RT), 100% removal was achieved. During continuous operation, the efficiency initially varied with the VBR concentration and RT, but gradually increased in one to two days. Furthermore, the acute toxicity of the effluent reduced by a factor of 21.25-49.61, indicating that the PBB can be successfully operated under turbulent environmental conditions. VBR degradation involved stepwise demethylation and yielded partially dealkylated VBR species. Phylogenetic analysis showed that the dominant phylum in the PBB was Proteobacteria and that Aeromonas hydrophila dominated during the entire operating period. The characteristics of the identified species showed that the PBB is suitable for processes such as demethylation, aromatic ring opening, carbon oxidation, nitrification, and denitrification. PMID:26657084

  6. Biodegradation of low-ethoxylated nonylphenols in a bioreactor packed with a new ceramic support (Vukopor ® S10).

    PubMed

    Sciubba, Luigi; Bertin, Lorenzo; Todaro, Daniela; Bettini, Cristina; Fava, Fabio; Di Gioia, Diana

    2014-03-01

    This work was aimed at studying the possibility of biodegrading 4-nonylphenol and low ethoxylated nonylphenol mixtures, which are particularly recalcitrant to microbial degradation, by employing a biofilm reactor packed with a ceramic support (Vukopor® S10). A selected microbial consortium (Consortium A) was used to colonize the support. 4-Nonylphenol and ethoxylated nonylphenol degradation and mineralization capabilities were studied both in batch and continuous mode. The results showed that Vukopor® S10 was able to be colonized by an active biofilm for the degradation of the target pollutants with the reactor operating both in batch and continuous mode. On the other hand, pollutant adsorption on the support was negligible. FISH showed equal proportion of Alphaproteobacteria and Gammaproteobacteria in the Igepal CO-520 degrading reactor. A shift towards high proportion of Gammaproteobacteria was observed by supplying Igepal CO-210. PCR-density gradient gel electrophoresis (DGGE) analyses also evidenced that the biofilm evolved with time by changing the mixture applied and that Proteobacteria were the most represented phylum in the biofilm. Taken together, the data obtained provide a strong indication that the biofilm reactor packed with Vukopor® S10 and inoculated with Consortium A could potentially be used to develop a technology for the decontamination of 4-nonylphenol and low ethoxylated nonylphenol polluted effluents.

  7. Biocatalytic methanation of hydrogen and carbon dioxide in a fixed bed bioreactor.

    PubMed

    Alitalo, Anni; Niskanen, Marko; Aura, Erkki

    2015-11-01

    Biocatalytic methanation of H2 and CO2 was studied in a fixed bed reactor system consisting of two solid state bioreactors in series connected to a recirculation system. Bioreactors were packed with a mixture of vermiculite shales and granular perlite material as a support material. A maximal methane productivity of 6.35l/lreactord was achieved at a hydrogen feed rate of 25.2l/lreactord, while hydrogen conversion rate was 100%. However, stable operation of the reactor at this efficiency remains to be achieved. Very simple reactor design, constructed from low cost materials, and the idea of exploiting waste material as a robust source of nutrients for methanogens makes this study very interesting regarding the overall usability and suitability of the system as part of a decentralized energy system. PMID:26298404

  8. Human embryonic stem cell-derived mesodermal progenitors display substantially increased tissue formation compared to human mesenchymal stem cells under dynamic culture conditions in a packed bed/column bioreactor.

    PubMed

    de Peppo, Giuseppe Maria; Sladkova, Martina; Sjövall, Peter; Palmquist, Anders; Oudina, Karim; Hyllner, Johan; Thomsen, Peter; Petite, Hervé; Karlsson, Camilla

    2013-01-01

    Bone tissue engineering represents a promising strategy to obviate bone deficiencies, allowing the ex vivo construction of bone substitutes with unprecedented potential in the clinical practice. Considering that in the human body cells are constantly stimulated by chemical and mechanical stimuli, the use of bioreactor is emerging as an essential factor for providing the proper environment for the reproducible and large-scale production of the engineered substitutes. Human mesenchymal stem cells (hMSCs) are experimentally relevant cells but, regardless the encouraging results reported after culture under dynamic conditions in bioreactors, show important limitations for tissue engineering applications, especially considering their limited proliferative potential, loss of functionality following protracted expansion, and decline in cellular fitness associated with aging. On the other hand, we previously demonstrated that human embryonic stem cell-derived mesodermal progenitors (hES-MPs) hold great potential to provide a homogenous and unlimited source of cells for bone engineering applications. Based on prior scientific evidence using different types of stem cells, in the present study we hypothesized that dynamic culture of hES-MPs in a packed bed/column bioreactor had the potential to affect proliferation, expression of genes involved in osteogenic differentiation, and matrix mineralization, therefore resulting in increased bone-like tissue formation. The reported findings suggest that hES-MPs constitute a suitable alternative cell source to hMSCs and hold great potential for the construction of bone substitutes for tissue engineering applications in clinical settings.

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

    SciTech Connect

    Hwang, P.; Chow, T.; Adrian, N.R.

    2000-04-01

    2,4,6-Trinitrotoluene (TNT) is an explosive widely used by the military. Although it is no longer manufactured in the US, large amounts of wastewater are generated annually from load, assembly, packing, and demilitarization operations. Granular-activated carbon adsorption is the standard technology for treating wastewater containing TNT and maintaining discharges within the limits established under the National Pollutant Discharge Elimination System. Studies evaluating biological treatment of pink water with an anaerobic fluidized-bed, granular-activated carbon bioreactor have been promising, but the fate of TNT is unknown. The authors investigated the anaerobic transformation of TNT by biofilm microorganisms obtained from a wastewater treatment plant receiving explosive manufacturing wastewater. The TNT was transformed to a mixture of 2-amino-4,6-dinitrotoluene; 4-amino-2,6-dinitrotoluene; 2,4-diamino-6-nitrotoluene; and 2,6-diamino-4-nitrotoluene before culminating in the formation of triaminotoluene (TAT). Triaminotoluene was susceptible to further degradation under anaerobic conditions, but its fate was not determined. Methane formation was inhibited but resumed after the depletion of the diaminonitrotoluene isomers. These studies demonstrate near stoichiometric formation of TAT from TNT and the transformation of 2-amino-4,6-dinitrotoluene to 2,4-diamino-6-nitrotoluene and 2,6-diamino-4-nitrotoluene by a mixed culture incubated under methanogenic conditions. This evidence indicates TAT is also a likely end-product of TNT biodegradation in the anaerobic fluidized fed bioreactor.

  10. Osmoregulation in methanogens

    SciTech Connect

    Roberts, M.F.

    1993-01-01

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

  11. NASA Bioreactor

    NASA Technical Reports Server (NTRS)

    1996-01-01

    Laptop computer sits atop the Experiment Control Computer for a NASA Bioreactor. The flight crew can change operating conditions in the Bioreactor by using the graphical interface on the laptop. The NASA Bioreactor provides a low turbulence culture environment which promotes the formation of large, three-dimensional cell clusters. The Bioreactor is rotated to provide gentle mixing of fresh and spent nutrient without inducing shear forces that would damage the cells. Due to their high level of cellular organization and specialization, samples constructed in the bioreactor more closely resemble the original tumor or tissue found in the body. The work is sponsored by NASA's Office of Biological and Physical Research. The bioreactor is managed by the Biotechnology Cell Science Program at NASA's Johnson Space Center (JSC). NASA-sponsored bioreactor research has been instrumental in helping scientists to better understand normal and cancerous tissue development. In cooperation with the medical community, the bioreactor design is being used to prepare better models of human colon, prostate, breast and ovarian tumors. Cartilage, bone marrow, heart muscle, skeletal muscle, pancreatic islet cells, liver and kidney are just a few of the normal tissues being cultured in rotating bioreactors by investigators.

  12. NASA Bioreactor

    NASA Technical Reports Server (NTRS)

    1996-01-01

    Close-up view of the interior of a NASA Bioreactor shows the plastic plumbing and valves (cylinders at center) to control fluid flow. A fresh nutrient bag is installed at top; a flattened waste bag behind it will fill as the nutrients are consumed during the course of operation. The drive chain and gears for the rotating wall vessel are visible at bottom center center. The NASA Bioreactor provides a low turbulence culture environment which promotes the formation of large, three-dimensional cell clusters. The Bioreactor is rotated to provide gentle mixing of fresh and spent nutrient without inducing shear forces that would damage the cells. Due to their high level of cellular organization and specialization, samples constructed in the bioreactor more closely resemble the original tumor or tissue found in the body. The work is sponsored by NASA's Office of Biological and Physical Research. The bioreactor is managed by the Biotechnology Cell Science Program at NASA's Johnson Space Center (JSC). NASA-sponsored bioreactor research has been instrumental in helping scientists to better understand normal and cancerous tissue development. In cooperation with the medical community, the bioreactor design is being used to prepare better models of human colon, prostate, breast and ovarian tumors. Cartilage, bone marrow, heart muscle, skeletal muscle, pancreatic islet cells, liver and kidney are just a few of the normal tissues being cultured in rotating bioreactors by investigators.

  13. NASA Bioreactor

    NASA Technical Reports Server (NTRS)

    1996-01-01

    Interior view of the gas supply for the NASA Bioreactor. The NASA Bioreactor provides a low turbulence culture environment which promotes the formation of large, three-dimensional cell clusters. The Bioreactor is rotated to provide gentle mixing of fresh and spent nutrient without inducing shear forces that would damage the cells. Due to their high level of cellular organization and specialization, samples constructed in the bioreactor more closely resemble the original tumor or tissue found in the body. The work is sponsored by NASA's Office of Biological and Physical Research. The bioreactor is managed by the Biotechnology Cell Science Program at NASA's Johnson Space Center (JSC). NASA-sponsored bioreactor research has been instrumental in helping scientists to better understand normal and cancerous tissue development. In cooperation with the medical community, the bioreactor design is being used to prepare better models of human colon, prostate, breast and ovarian tumors. Cartilage, bone marrow, heart muscle, skeletal muscle, pancreatic islet cells, liver and kidney are just a few of the normal tissues being cultured in rotating bioreactors by investigators.

  14. NASA Bioreactor

    NASA Technical Reports Server (NTRS)

    1996-01-01

    Biotechnology Refrigerator that preserves samples for use in (or after culturing in) the NASA Bioreactor. The unit is shown extracted from a middeck locker shell. The NASA Bioreactor provides a low turbulence culture environment which promotes the formation of large, three-dimensional cell clusters. The Bioreactor is rotated to provide gentle mixing of fresh and spent nutrient without inducing shear forces that would damage the cells. Due to their high level of cellular organization and specialization, samples constructed in the bioreactor more closely resemble the original tumor or tissue found in the body. The work is sponsored by NASA's Office of Biological and Physical Research. The bioreactor is managed by the Biotechnology Cell Science Program at NASA's Johnson Space Center (JSC). NASA-sponsored bioreactor research has been instrumental in helping scientists to better understand normal and cancerous tissue development. In cooperation with the medical community, the bioreactor design is being used to prepare better models of human colon, prostate, breast and ovarian tumors. Cartilage, bone marrow, heart muscle, skeletal muscle, pancreatic islet cells, liver and kidney are just a few of the normal tissues being cultured in rotating bioreactors by investigators.

  15. NASA Bioreactor

    NASA Technical Reports Server (NTRS)

    1996-01-01

    Exterior view of the NASA Bioreactor Engineering Development Unit flown on Mir. The rotating wall vessel is behind the window on the face of the large module. Control electronics are in the module at left; gas supply and cooling fans are in the module at back. The NASA Bioreactor provides a low turbulence culture environment which promotes the formation of large, three-dimensional cell clusters. The Bioreactor is rotated to provide gentle mixing of fresh and spent nutrient without inducing shear forces that would damage the cells. Due to their high level of cellular organization and specialization, samples constructed in the bioreactor more closely resemble the original tumor or tissue found in the body. The work is sponsored by NASA's Office of Biological and Physical Research. The bioreactor is managed by the Biotechnology Cell Science Program at NASA's Johnson Space Center (JSC). NASA-sponsored bioreactor research has been instrumental in helping scientists to better understand normal and cancerous tissue development. In cooperation with the medical community, the bioreactor design is being used to prepare better models of human colon, prostate, breast and ovarian tumors. Cartilage, bone marrow, heart muscle, skeletal muscle, pancreatic islet cells, liver and kidney are just a few of the normal tissues being cultured in rotating bioreactors by investigators.

  16. NASA Bioreactor

    NASA Technical Reports Server (NTRS)

    1996-01-01

    Electronics control module for the NASA Bioreactor. The NASA Bioreactor provides a low turbulence culture environment which promotes the formation of large, three-dimensional cell clusters. The Bioreactor is rotated to provide gentle mixing of fresh and spent nutrient without inducing shear forces that would damage the cells. Due to their high level of cellular organization and specialization, samples constructed in the bioreactor more closely resemble the original tumor or tissue found in the body. The work is sponsored by NASA's Office of Biological and Physical Research. The bioreactor is managed by the Biotechnology Cell Science Program at NASA's Johnson Space Center (JSC). NASA-sponsored bioreactor research has been instrumental in helping scientists to better understand normal and cancerous tissue development. In cooperation with the medical community, the bioreactor design is being used to prepare better models of human colon, prostate, breast and ovarian tumors. Cartilage, bone marrow, heart muscle, skeletal muscle, pancreatic islet cells, liver and kidney are just a few of the normal tissues being cultured in rotating bioreactors by investigators.

  17. NASA Bioreactor

    NASA Technical Reports Server (NTRS)

    1996-01-01

    Interior of a Biotechnology Refrigerator that preserves samples for use in (or after culturing in) the NASA Bioreactor. The unit is shown extracted from a middeck locker shell. The NASA Bioreactor provides a low turbulence culture environment which promotes the formation of large, three-dimensional cell clusters. The Bioreactor is rotated to provide gentle mixing of fresh and spent nutrient without inducing shear forces that would damage the cells. Due to their high level of cellular organization and specialization, samples constructed in the bioreactor more closely resemble the original tumor or tissue found in the body. The work is sponsored by NASA's Office of Biological and Physical Research. The bioreactor is managed by the Biotechnology Cell Science Program at NASA's Johnson Space Center (JSC). NASA-sponsored bioreactor research has been instrumental in helping scientists to better understand normal and cancerous tissue development. In cooperation with the medical community, the bioreactor design is being used to prepare better models of human colon, prostate, breast and ovarian tumors. Cartilage, bone marrow, heart muscle, skeletal muscle, pancreatic islet cells, liver and kidney are just a few of the normal tissues being cultured in rotating bioreactors by investigators.

  18. NASA Bioreactor

    NASA Technical Reports Server (NTRS)

    1996-01-01

    Biotechnology Refrigerator that preserves samples for use in (or after culturing in) the NASA Bioreactor. The unit is shown extracted from a middeck locker shell and with thermal blankets partially removed. The NASA Bioreactor provides a low turbulence culture environment which promotes the formation of large, three-dimensional cell clusters. The Bioreactor is rotated to provide gentle mixing of fresh and spent nutrient without inducing shear forces that would damage the cells. Due to their high level of cellular organization and specialization, samples constructed in the bioreactor more closely resemble the original tumor or tissue found in the body. The work is sponsored by NASA's Office of Biological and Physical Research. The bioreactor is managed by the Biotechnology Cell Science Program at NASA's Johnson Space Center (JSC). NASA-sponsored bioreactor research has been instrumental in helping scientists to better understand normal and cancerous tissue development. In cooperation with the medical community, the bioreactor design is being used to prepare better models of human colon, prostate, breast and ovarian tumors. Cartilage, bone marrow, heart muscle, skeletal muscle, pancreatic islet cells, liver and kidney are just a few of the normal tissues being cultured in rotating bioreactors by investigators.

  19. NASA Bioreactor

    NASA Technical Reports Server (NTRS)

    1996-01-01

    Close-up view of the interior of a NASA Bioreactor shows the plastic plumbing and valves (cylinders at right center) to control fluid flow. The rotating wall vessel is at top center. The NASA Bioreactor provides a low turbulence culture environment which promotes the formation of large, three-dimensional cell clusters. The Bioreactor is rotated to provide gentle mixing of fresh and spent nutrient without inducing shear forces that would damage the cells. Due to their high level of cellular organization and specialization, samples constructed in the bioreactor more closely resemble the original tumor or tissue found in the body. The work is sponsored by NASA's Office of Biological and Physical Research. The bioreactor is managed by the Biotechnology Cell Science Program at NASA's Johnson Space Center (JSC). NASA-sponsored bioreactor research has been instrumental in helping scientists to better understand normal and cancerous tissue development. In cooperation with the medical community, the bioreactor design is being used to prepare better models of human colon, prostate, breast and ovarian tumors. Cartilage, bone marrow, heart muscle, skeletal muscle, pancreatic islet cells, liver and kidney are just a few of the normal tissues being cultured in rotating bioreactors by investigators.

  20. NASA Bioreactor

    NASA Technical Reports Server (NTRS)

    1998-01-01

    Biotechnology Refrigerator (BTR) holds fixed tissue culture bags at 4 degrees C to preserve them for return to Earth and postflight analysis. The cultures are used in research with the NASA Bioreactor cell science program. The work is sponsored by NASA's Office of Biological and Physical Research. The bioreactor is managed by the Biotechnology Cell Science Program at NASA's Johnson Space Center (JSC).

  1. NASA Bioreactor

    NASA Technical Reports Server (NTRS)

    1998-01-01

    The heart of the bioreactor is the rotating wall vessel, shown without its support equipment. Volume is about 125 mL. The work is sponsored by NASA's Office of Biological and Physical Research. The bioreactor is managed by the Biotechnology Cell Science Program at NASA's Johnson Space Center (JSC). NASA-sponsored bioreactor research has been instrumental in helping scientists to better understand normal and cancerous tissue development. In cooperation with the medical community, the bioreactor design is being used to prepare better models of human colon, prostate, breast and ovarian tumors. Cartilage, bone marrow, heart muscle, skeletal muscle, pancreatic islet cells, liver and kidney are just a few of the normal tissues being cultured in rotating bioreactors by investigators.

  2. NASA Bioreactor

    NASA Technical Reports Server (NTRS)

    1998-01-01

    Astronaut John Blaha replaces an exhausted media bag and filled waste bag with fresh bags to continue a bioreactor experiment aboard space station Mir in 1996. NASA-sponsored bioreactor research has been instrumental in helping scientists to better understand normal and cancerous tissue development. In cooperation with the medical community, the bioreactor design is being used to prepare better models of human colon, prostate, breast and ovarian tumors. Cartilage, bone marrow, heart muscle, skeletal muscle, pancreatic islet cells, liver and kidney are just a few of the normal tissues being cultured in rotating bioreactors by investigators. This image is from a video downlink. The work is sponsored by NASA's Office of Biological and Physical Research. The bioreactor is managed by the Biotechnology Cell Science Program at NASA's Johnson Space Center (JSC).

  3. Bioreactor principles

    NASA Technical Reports Server (NTRS)

    2001-01-01

    Cells cultured on Earth (left) typically settle quickly on the bottom of culture vessels due to gravity. In microgravity (right), cells remain suspended and aggregate to form three-dimensional tissue. The NASA Bioreactor provides a low turbulence culture environment which promotes the formation of large, three-dimensional cell clusters. The Bioreactor is rotated to provide gentle mixing of fresh and spent nutrient without inducing shear forces that would damage the cells. Due to their high level of cellular organization and specialization, samples constructed in the bioreactor more closely resemble the original tumor or tissue found in the body. The work is sponsored by NASA's Office of Biological and Physical Research. The bioreactor is managed by the Biotechnology Cell Science Program at NASA's Johnson Space Center (JSC). NASA-sponsored bioreactor research has been instrumental in helping scientists to better understand normal and cancerous tissue development. In cooperation with the medical community, the bioreactor design is being used to prepare better models of human colon, prostate, breast and ovarian tumors. Cartilage, bone marrow, heart muscle, skeletal muscle, pancreatic islet cells, liver and kidney are just a few of the normal tissues being cultured in rotating bioreactors by investigators.

  4. Nickel isotopes and methanogens

    NASA Astrophysics Data System (ADS)

    Neubeck, A.; Ivarsson, M.

    2013-12-01

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

  5. Adaptation of a methanogenic consortium to arsenite inhibition

    PubMed Central

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

    2016-01-01

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

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

    SciTech Connect

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

    2010-08-05

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

  7. NASA Bioreactor

    NASA Technical Reports Server (NTRS)

    1998-01-01

    Biotechnology Specimen Temperature Controller (BSTC) will cultivate cells until their turn in the bioreactor; it can also be used in culturing experiments that do not require the bioreactor. The BSTC comprises four incubation/refrigeration chambers individually set at 4 to 50 deg. C (near-freezing to above body temperature). Each chamber holds three rugged tissue chamber modules (12 total), clear Teflon bags holding 30 ml of growth media, all positioned by a metal frame. Every 7 to 21 days (depending on growth rates), an astronaut uses a shrouded syringe and the bags' needleless injection ports to transfer a few cells to a fresh media bag, and to introduce a fixative so that the cells may be studied after flight. The design also lets the crew sample the media to measure glucose, gas, and pH levels, and to inspect cells with a microscope. The controller is monitored by the flight crew through a 23-cm (9-inch) color computer display on the face of the BSTC. This view shows the BTSC with the front panel open. The work is sponsored by NASA's Office of Biological and Physical Research. The bioreactor is managed by the Biotechnology Cell Science Program at NASA's Johnson Space Center (JSC). NASA-sponsored bioreactor research has been instrumental in helping scientists to better understand normal and cancerous tissue development. In cooperation with the medical community, the bioreactor design is being used to prepare better models of human colon, prostate, breast and ovarian tumors. Cartilage, bone marrow, heart muscle, skeletal muscle, pancreatic islet cells, liver and kidney are just a few of the normal tissues being cultured in rotating bioreactors by investigators.

  8. NASA Bioreactor

    NASA Technical Reports Server (NTRS)

    1998-01-01

    Bioreactor Demonstration System (BDS) comprises an electronics module, a gas supply module, and the incubator module housing the rotating wall vessel and its support systems. Nutrient media are pumped through an oxygenator and the culture vessel. The shell rotates at 0.5 rpm while the irner filter typically rotates at 11.5 rpm to produce a gentle flow that ensures removal of waste products as fresh media are infused. Periodically, some spent media are pumped into a waste bag and replaced by fresh media. When the waste bag is filled, an astronaut drains the waste bag and refills the supply bag through ports on the face of the incubator. Pinch valves and a perfusion pump ensure that no media are exposed to moving parts. An Experiment Control Computer controls the Bioreactor, records conditions, and alerts the crew when problems occur. The crew operates the system through a laptop computer displaying graphics designed for easy crew training and operation. The work is sponsored by NASA's Office of Biological and Physical Research. The bioreactor is managed by the Biotechnology Cell Science Program at NASA's Johnson Space Center (JSC). NASA-sponsored bioreactor research has been instrumental in helping scientists to better understand normal and cancerous tissue development. In cooperation with the medical community, the bioreactor design is being used to prepare better models of human colon, prostate, breast and ovarian tumors. Cartilage, bone marrow, heart muscle, skeletal muscle, pancreatic islet cells, liver and kidney are just a few of the normal tissues being cultured in rotating bioreactors by investigators. See No. 0101824 for a version with labels, and No. 0103180 for an operational schematic.

  9. NASA Bioreactor

    NASA Technical Reports Server (NTRS)

    1998-01-01

    Bioreactor Demonstration System (BDS) comprises an electronics module, a gas supply module, and the incubator module housing the rotating wall vessel and its support systems. Nutrient media are pumped through an oxygenator and the culture vessel. The shell rotates at 0.5 rpm while the irner filter typically rotates at 11.5 rpm to produce a gentle flow that ensures removal of waste products as fresh media are infused. Periodically, some spent media are pumped into a waste bag and replaced by fresh media. When the waste bag is filled, an astronaut drains the waste bag and refills the supply bag through ports on the face of the incubator. Pinch valves and a perfusion pump ensure that no media are exposed to moving parts. An Experiment Control Computer controls the Bioreactor, records conditions, and alerts the crew when problems occur. The crew operates the system through a laptop computer displaying graphics designed for easy crew training and operation. The work is sponsored by NASA's Office of Biological and Physical Research. The bioreactor is managed by the Biotechnology Cell Science Program at NASA's Johnson Space Center (JSC). NASA-sponsored bioreactor research has been instrumental in helping scientists to better understand normal and cancerous tissue development. In cooperation with the medical community, the bioreactor design is being used to prepare better models of human colon, prostate, breast and ovarian tumors. Cartilage, bone marrow, heart muscle, skeletal muscle, pancreatic islet cells, liver and kidney are just a few of the normal tissues being cultured in rotating bioreactors by investigators. See No. 0101816 for a version without labels, and No. 0103180 for an operational schematic.

  10. NASA Bioreactor

    NASA Technical Reports Server (NTRS)

    1998-01-01

    Bioreactor Demonstration System (BDS) comprises an electronics module, a gas supply module, and the incubator module housing the rotating wall vessel and its support systems. Nutrient media are pumped through an oxygenator and the culture vessel. The shell rotates at 0.5 rpm while the irner filter typically rotates at 11.5 rpm to produce a gentle flow that ensures removal of waste products as fresh media are infused. Periodically, some spent media are pumped into a waste bag and replaced by fresh media. When the waste bag is filled, an astronaut drains the waste bag and refills the supply bag through ports on the face of the incubator. Pinch valves and a perfusion pump ensure that no media are exposed to moving parts. An Experiment Control Computer controls the Bioreactor, records conditions, and alerts the crew when problems occur. The crew operates the system through a laptop computer displaying graphics designed for easy crew training and operation. The work is sponsored by NASA's Office of Biological and Physical Research. The bioreactor is managed by the Biotechnology Cell Science Program at NASA's Johnson Space Center (JSC). NASA-sponsored bioreactor research has been instrumental in helping scientists to better understand normal and cancerous tissue development. In cooperation with the medical community, the bioreactor design is being used to prepare better models of human colon, prostate, breast and ovarian tumors. Cartilage, bone marrow, heart muscle, skeletal muscle, pancreatic islet cells, liver and kidney are just a few of the normal tissues being cultured in rotating bioreactors by investigators. See No. 0101823 for a version without labels, and No. 0103180 for an operational schematic.

  11. NASA Bioreactor

    NASA Technical Reports Server (NTRS)

    1998-01-01

    Bioreactor Demonstration System (BDS) comprises an electronics module, a gas supply module, and the incubator module housing the rotating wall vessel and its support systems. Nutrient media are pumped through an oxygenator and the culture vessel. The shell rotates at 0.5 rpm while the irner filter typically rotates at 11.5 rpm to produce a gentle flow that ensures removal of waste products as fresh media are infused. Periodically, some spent media are pumped into a waste bag and replaced by fresh media. When the waste bag is filled, an astronaut drains the waste bag and refills the supply bag through ports on the face of the incubator. Pinch valves and a perfusion pump ensure that no media are exposed to moving parts. An Experiment Control Computer controls the Bioreactor, records conditions, and alerts the crew when problems occur. The crew operates the system through a laptop computer displaying graphics designed for easy crew training and operation. The work is sponsored by NASA's Office of Biological and Physical Research. The bioreactor is managed by the Biotechnology Cell Science Program at NASA's Johnson Space Center (JSC). NASA-sponsored bioreactor research has been instrumental in helping scientists to better understand normal and cancerous tissue development. In cooperation with the medical community, the bioreactor design is being used to prepare better models of human colon, prostate, breast and ovarian tumors. Cartilage, bone marrow, heart muscle, skeletal muscle, pancreatic islet cells, liver and kidney are just a few of the normal tissues being cultured in rotating bioreactors by investigators. See No. 0101825 for a version with major elements labeled, and No. 0103180 for an operational schematic. 0101816

  12. NASA Bioreactor

    NASA Technical Reports Server (NTRS)

    1998-01-01

    Biotechnology Specimen Temperature Controller (BSTC) will cultivate cells until their turn in the bioreactor; it can also be used in culturing experiments that do not require the bioreactor. The BSTC comprises four incubation/refrigeration chambers individually set at 4 to 50 degreesC (near-freezing to above body temperature). Each chamber holds three rugged tissue chamber modules (12 total), clear Teflon bags holding 30 ml of growth media, all positioned by a metal frame. Every 7 to 21 days (depending on growth rates), an astronaut uses a shrouded syringe and the bags' needleless injection ports to transfer a few cells to a fresh media bag, and to introduce a fixative so that the cells may be studied after flight. The design also lets the crew sample the media to measure glucose, gas, and pH levels, and to inspect cells with a microscope. The controller is monitored by the flight crew through a 23-cm (9-inch) color computer display on the face of the BSTC. This view shows the BTSC with the front panel open. The work is sponsored by NASA's Office of Biological and Physical Research. The bioreactor is managed by the Biotechnology Cell Science Program at NASA's Johnson Space Center (JSC). NASA-sponsored bioreactor research has been instrumental in helping scientists to better understand normal and cancerous tissue development. In cooperation with the medical community, the bioreactor design is being used to prepare better models of human colon, prostate, breast and ovarian tumors. Cartilage, bone marrow, heart muscle, skeletal muscle, pancreatic islet cells, liver and kidney are just a few of the normal tissues being cultured in rotating bioreactors by investigators.

  13. Multimembrane Bioreactor

    NASA Technical Reports Server (NTRS)

    Cho, Toohyon; Shuler, Michael L.

    1989-01-01

    Set of hydrophilic and hydrophobic membranes in bioreactor allows product of reaction to be separated, while nutrients fed to reacting cells and byproducts removed from them. Separation process requires no externally supplied energy; free energy of reaction sufficient. Membranes greatly increase productivity of metabolizing cells by continuously removing product and byproducts, which might otherwise inhibit reaction, and by continuously adding oxygen and organic nutrients.

  14. Light sensitivity of methanogenic archaebacteria

    SciTech Connect

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

    1991-09-01

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

  15. Advanced bioreactor concepts for coal processing

    SciTech Connect

    Scott, C.D.

    1988-01-01

    The development of advanced bioreactor systems for the processing of coal should follow some basic principles. Continuous operation is preferred, with maximum bioreagent concentrations and enhanced mass transfer. Although conventional stirred-tank bioreactors will be more appropriate for some processing concepts, columnar reactors with retained bioreagents could be the system of choice for most of the applications. Serious consideration must now be given to process development of some biological coal processing concepts. Process biology and biochemistry will continue to be very important, but efficient bioreactor systems will be necessary for economic feasibility. Conventional bioreactor concepts will be useful for some applications, but columnar systems represent an innovative approach to the design of continuous bioreactors with high productivity and good operational control. Fluidized and packed beds are the most promising configurations, especially where three-phase operation is required and where interphase mass transport is a likely controlling mechanism. Although the biocatalyst must be immobilized into or onto particles to be retained in the bioreactors, this also results in a very high biocatalyst concentration without washout and a significant enhancement in bioconversion rates. The multistage nature of these types of bioreactors also contributes to higher efficiencies for many types of biocatalytic processes. 25 refs.

  16. The Geobiochemistry of Methanogen Proteins

    NASA Astrophysics Data System (ADS)

    Prasad, A.; Shock, E.

    2013-12-01

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

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

    SciTech Connect

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

    1999-10-01

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

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

    USGS Publications Warehouse

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

    1999-01-01

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

  19. Bioreactors Addressing Diabetes Mellitus

    PubMed Central

    Minteer, Danielle M.; Gerlach, Jorg C.

    2014-01-01

    The concept of bioreactors in biochemical engineering is a well-established process; however, the idea of applying bioreactor technology to biomedical and tissue engineering issues is relatively novel and has been rapidly accepted as a culture model. Tissue engineers have developed and adapted various types of bioreactors in which to culture many different cell types and therapies addressing several diseases, including diabetes mellitus types 1 and 2. With a rising world of bioreactor development and an ever increasing diagnosis rate of diabetes, this review aims to highlight bioreactor history and emerging bioreactor technologies used for diabetes-related cell culture and therapies. PMID:25160666

  20. Bioreactors addressing diabetes mellitus.

    PubMed

    Minteer, Danielle M; Gerlach, Jorg C; Marra, Kacey G

    2014-11-01

    The concept of bioreactors in biochemical engineering is a well-established process; however, the idea of applying bioreactor technology to biomedical and tissue engineering issues is relatively novel and has been rapidly accepted as a culture model. Tissue engineers have developed and adapted various types of bioreactors in which to culture many different cell types and therapies addressing several diseases, including diabetes mellitus types 1 and 2. With a rising world of bioreactor development and an ever increasing diagnosis rate of diabetes, this review aims to highlight bioreactor history and emerging bioreactor technologies used for diabetes-related cell culture and therapies.

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

    SciTech Connect

    Xu, Qiyong; Tian, Ying; Wang, Shen; Ko, Jae Hac

    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.

  2. Optimization of denitrifying bioreactor performance with agricultural residue-based filter media

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Denitrification bioreactors are a promising technology for mitigation of nitrate-nitrogen (NO3-N) losses in subsurface drainage water. Bioreactors are constructed with carbon substrates, typically wood chips, to provide a substrate for denitrifying microorganisms. Columns were packed with wood chips...

  3. Bio-reactor chamber

    NASA Technical Reports Server (NTRS)

    Chandler, Joseph A. (Inventor)

    1989-01-01

    A bioreactor for cell culture is disclosed which provides for the introduction of fresh medium without excessive turbulent action. The fresh medium enters the bioreactor through a filter with a backwash action which prevents the cells from settling on the filter. The bioreactor is sealed and depleted medium is forced out of the container as fresh medium is added.

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

    PubMed

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

    2016-02-01

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

  5. Methanogens in the Solar System

    NASA Astrophysics Data System (ADS)

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

    2015-04-01

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

  6. Methanogenic activities in alpine soils.

    PubMed

    Wagner, Andreas O; Hofmann, Katrin; Prem, Eva; Illmer, Paul

    2012-07-01

    Uncontrolled microbial methane production is playing an important role in global warming. In the present study, we showed that water content and incubation temperature increase the potential for methane formation in the two alpine soils under investigation. Beside these factors, the grazing of cows and thus the amendment of methanogenic microorganisms by cattle dung is the most important factor determining the potential of methane production in those soils.

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

    PubMed

    Xu, Qiyong; Tian, Ying; Wang, Shen; Ko, Jae Hac

    2015-07-01

    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 10months. 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 75d and 60d 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 133.4L/kgvs and 113.2L/kgvs. As for MSW with high content of food waste, leachate recirculation right after aeration stopped was not recommended due to VFA inhibition for methanogens.

  8. Bioreactors for tissue engineering.

    PubMed

    Chen, Huang-Chi; Hu, Yu-Chen

    2006-09-01

    Bioreactors are essential in tissue engineering, not only because they provide an in vitro environment mimicking in vivo conditions for the growth of tissue substitutes, but also because they enable systematic studies of the responses of living tissues to various mechanical and biochemical cues. The basic principles of bioreactor design are reviewed, the bioreactors commonly used for the tissue engineering of cartilage, bone and cardiovascular systems are assessed in terms of their performance and usefulness. Several novel bioreactor types are also reviewed. PMID:16955350

  9. Disposable polymeric cryogel bioreactor matrix for therapeutic protein production.

    PubMed

    Jain, Era; Kumar, Ashok

    2013-05-01

    Low cost and high efficiency make disposable bioreactors feasible for small-scale therapeutic development and initial clinical trials. We have developed a cryogel-based disposable bioreactor matrix, which has been used for production of protein therapeutics such as urokinase and monoclonal antibodies (mAbs). The protocol discusses the application of a cryogel bioreactor for mAb production. Cryogels composed of either polyacrylamide (PAAm) coupled to gelatin or semi-interpenetrating PAAm-chitosan are synthesized by free-radical polymerization at -12 °C. Hybridoma cells are immobilized over the cryogel bioreactor and incubated for 48 h. Medium is circulated thereafter at 0.2 ml min(-1) and bioreactors can be run continuously for 60 d. The cryogel-based packed-bed bioreactor can be formulated as a monolith or as beads; it also has an efficiency four times what can be obtained using a tissue-culture flask, a high surface-to-volume ratio and effective nutrient transport. After incubation, the bioreactor setup will take about 60 min using a pre-prepared sterilized cryogel.

  10. NASA Bioreactor Demonstration System

    NASA Technical Reports Server (NTRS)

    2002-01-01

    Leland W. K. Chung (left), Director, Molecular Urology Therapeutics Program at the Winship Cancer Institute at Emory University, is principal investigator for the NASA bioreactor demonstration system (BDS-05). With him is Dr. Jun Shu, an assistant professor of Orthopedics Surgery from Kuming Medical University China. The NASA Bioreactor provides a low turbulence culture environment which promotes the formation of large, three-dimensional cell clusters. Due to their high level of cellular organization and specialization, samples constructed in the bioreactor more closely resemble the original tumor or tissue found in the body. The Bioreactor is rotated to provide gentle mixing of fresh and spent nutrient without inducing shear forces that would damage the cells. The work is sponsored by NASA's Office of Biological and Physical Research. The bioreactor is managed by the Biotechnology Cell Science Program at NASA's Johnson Space Center (JSC). NASA-sponsored bioreactor research has been instrumental in helping scientists to better understand normal and cancerous tissue development. In cooperation with the medical community, the bioreactor design is being used to prepare better models of human colon, prostate, breast and ovarian tumors. Cartilage, bone marrow, heart muscle, skeletal muscle, pancreatic islet cells, liver and kidney are just a few of the normal tissues being cultured in rotating bioreactors by investigators. Credit: Emory University.

  11. Modularized evolution in archaeal methanogens phylogenetic forest.

    PubMed

    Li, Jun; Wong, Chi-Fat; Wong, Mabel Ting; Huang, He; Leung, Frederick C

    2014-12-09

    Methanogens are methane-producing archaea that plays a key role in the global carbon cycle. To date, the evolutionary history of methanogens and closely related nonmethanogen species remains unresolved among studies conducted upon different genetic markers, attributing to horizontal gene transfers (HGTs). With an effort to decipher both congruent and conflicting evolutionary events, reconstruction of coevolved gene clusters and hierarchical structure in the archaeal methanogen phylogenetic forest, comprehensive evolution, and network analyses were performed upon 3,694 gene families from 41 methanogens and 33 closely related archaea. Our results show that 1) greater than 50% of genes are in topological dissonance with others; 2) the prevalent interorder HGTs, even for core genes, in methanogen genomes led to their scrambled phylogenetic relationships; 3) most methanogenesis-related genes have experienced at least one HGT; 4) greater than 20% of the genes in methanogen genomes were transferred horizontally from other archaea, with genes involved in cell-wall synthesis and defense system having been transferred most frequently; 5) the coevolution network contains seven statistically robust modules, wherein the central module has the highest average node strength and comprises a majority of the core genes; 6) different coevolutionary module genes boomed in different time and evolutionary lineage, constructing diversified pan-genome structures; 7) the modularized evolution is also closely related to the vertical evolution signals and the HGT rate of the genes. Overall, this study presented a modularized phylogenetic forest that describes a combination of complicated vertical and nonvertical evolutionary processes for methanogenic archaeal species.

  12. Hepatocyte function within a stacked double sandwich culture plate cylindrical bioreactor for bioartificial liver system.

    PubMed

    Xia, Lei; Arooz, Talha; Zhang, Shufang; Tuo, Xiaoye; Xiao, Guangfa; Susanto, Thomas Adi Kurnia; Sundararajan, Janani; Cheng, Tianming; Kang, Yuzhan; Poh, Hee Joo; Leo, Hwa Liang; Yu, Hanry

    2012-11-01

    Bioartificial liver (BAL) system is promising as an alternative treatment for liver failure. We have developed a bioreactor with stacked sandwich culture plates for the application of BAL. This bioreactor design addresses some of the persistent problems in flat-bed bioreactors through increasing cell packing capacity, eliminating dead flow, regulating shear stress, and facilitating the scalability of the bioreactor unit. The bioreactor contained a stack of twelve double-sandwich-culture plates, allowing 100 million hepatocytes to be housed in a single cylindrical bioreactor unit (7 cm of height and 5.5 cm of inner diameter). The serial flow perfusion through the bioreactor increased cell-fluid contact area for effective mass exchange. With the optimal perfusion flow rate, shear stress was minimized to achieve high and uniform cell viabilities across different plates in the bioreactor. Our results demonstrated that hepatocytes cultured in the bioreactor could re-establish cell polarity and maintain liver-specific functions (e.g. albumin and urea synthesis, phase I&II metabolism functions) for seven days. The single bioreactor unit can be readily scaled up to house adequate number of functional hepatocytes for BAL development.

  13. Relationship between methanogenic cofactor content and maximum specific methanogenic activity of anaerobic granular sludges

    SciTech Connect

    Gorris, L.G.; de Kok, T.M.; Kroon, B.M.; van der Drift, C.; Vogels, G.D.

    1988-05-01

    In this study we investigated whether a relationship exists between the methanogenic activity and the content of specific methanogenic cofactors of granular sludges cultured on different combinations of volatile fatty acids in upflow anaerobic sludge blanket or fluidized-bed reactors. Significant correlations were measured in both cases between the contents of coenzyme F/sub 420/-2 or methanopterin and the maximum specific methanogenic activities on propionate, butyrate, and hydrogen, but not acetate. For both sludges the content of sarcinapterin appeared to be correlated with methanogenic activities on propionate, butyrate, and acetate, but not hydrogen. Similar correlations were measured with regard to the total content of coenzyme F/sub 420/-4 and F/sub 420/-5 sludges from fluidized-bed reactors. The results indicate that the contents of specific methanogenic cofactors measured in anaerobic granular sludges can be used to estimate the hydrogenotrophic or acetotrophic methanogenic potential of these sludges.

  14. Bioreactor rotating wall vessel

    NASA Technical Reports Server (NTRS)

    2001-01-01

    The NASA Bioreactor provides a low turbulence culture environment which promotes the formation of large, three-dimensional cell clusters. Due to their high level of cellular organization and specialization, samples constructed in the bioreactor more closely resemble the original tumor or tissue found in the body. The work is sponsored by NASA's Office of Biological and Physical Research. The bioreactor is managed by the Biotechnology Cell Science Program at NASA's Johnson Space Center (JSC). NASA-sponsored bioreactor research has been instrumental in helping scientists to better understand normal and cancerous tissue development. In cooperation with the medical community, the bioreactor design is being used to prepare better models of human colon, prostate, breast and ovarian tumors. Cartilage, bone marrow, heart muscle, skeletal muscle, pancreatic islet cells, liver and kidney are just a few of the normal tissues being cultured in rotating bioreactors by investigators. Cell constructs grown in a rotating bioreactor on Earth (left) eventually become too large to stay suspended in the nutrient media. In the microgravity of orbit, the cells stay suspended. Rotation then is needed for gentle stirring to replenish the media around the cells.

  15. Configuration of bioreactors.

    PubMed

    Martens, Dirk E; van den End, Evert J; Streefland, Mathieu

    2014-01-01

    Lab-scale stirred-tank bioreactors (0.2-20 l) are used for fundamental research on animal cells and in process development and troubleshooting for large-scale production. In this chapter, different configurations of bioreactor systems are shortly discussed and setting up these different configurations is described. In addition, online measurement and control of bioreactor parameters is described, with special attention to controller settings (PID) and online measurement of oxygen consumption and carbon dioxide production. Finally, methods for determining the oxygen transfer coefficient are described.

  16. Bioreactors: design and operation

    SciTech Connect

    Cooney, C.L.

    1983-02-11

    The bioreactor provides a central link between the starting feedstock and the product. The reaction yield and selectivity are determined by the biocatalyst, but productivity is often determined by the process technology; as a consequence, biochemical reaction engineering becomes the interface for the biologist and engineer. Developments in bioreactor design, including whole cell immobilization, immobilized enzymes, continuous reaction, and process control, will increasingly reflect the need for cross-disciplinary interaction in the biochemical process industry. This paper examines the strategy for selection and design of bioreactors and identifies the limits and constraints in their use. 25 references, 3 figures, 3 tables.

  17. Hydrogenotrophic control in methanogenic processes

    SciTech Connect

    Smith, D.P.

    1987-01-01

    Interspecies transfer of molecular hydrogen from obligately proton-reducing or facultative fermenting bacteria to chemolithotrophic methanogens is an important control process in methanogenesis. Experimental studies are presented which elucidate these interactions, and demonstrate the significance of hydrogenotrophs in process control. Steady-state hydrogen partial pressure was measured under sixteen conditions, with continuous stirred tank reactors, anaerobic baffled reactors, and an anaerobic filter, operated at various detention times on ethanol, propionate, and other substrates. Longer-chain n-carboxylic acids were formed by back reactions from acetate and propionate when hydrogen levels were elevated; reactions reversed direction when hydrogen levels fell and equilibrium shifted to favor beta-oxidations. N-propanol was formed from propionate by an ethanol-oxidation/propionate-reduction reactions, determined to be mediated by ethanol-oxidizers; n-propanol formation was enhanced by elevated propionate concentrations. Substrate utilization and product formation were consistent with reaction energetics. Substrates were not consumed when their utilization reactions had a positive free energy change, and changes in product and substrate concentrations were reversed with a shift in equilibrium of the corresponding reactions. Hydrogen levels exerted the most significant influence on the direction of equilibrium. Energetic analysis offers a predictive tool to restrictively characterize reactions and reaction patterns. Biological processes fostering biofilm growth had higher hydrogen turnover rates than processes tending towards dispersed-growth. Biofilm processes offer greater operational stability than dispersed-growth processes in treating hydrogen-producing substrates.

  18. Space Bioreactor Science Workshop

    NASA Technical Reports Server (NTRS)

    Morrison, Dennis R. (Editor)

    1987-01-01

    The first space bioreactor has been designed for microprocessor control, no gaseous headspace, circulation and resupply of culture medium, and a slow mixing in very low shear regimes. Various ground based bioreactors are being used to test reactor vessel design, on-line sensors, effects of shear, nutrient supply, and waste removal from continuous culture of human cells attached to microcarriers. The small (500 ml) bioreactor is being constructed for flight experiments in the Shuttle middeck to verify systems operation under microgravity conditions and to measure the efficiencies of mass transport, gas transfer, oxygen consumption, and control of low shear stress on cells. Applications of microcarrier cultures, development of the first space bioreactor flight system, shear and mixing effects on cells, process control, and methods to monitor cell metabolism and nutrient requirements are among the topics covered.

  19. Bioreactor design concepts

    NASA Technical Reports Server (NTRS)

    Bowie, William

    1987-01-01

    Two parallel lines of work are underway in the bioreactor laboratory. One of the efforts is devoted to the continued development and utilization of a laboratory research system. That system's design is intended to be fluid and dynamic. The sole purpose of such a device is to allow testing and development of equipment concepts and procedures. Some of the results of those processes are discussed. A second effort is designed to produce a flight-like bioreactor contained in a double middeck locker. The result of that effort has been to freeze a particular bioreactor design in order to allow fabrication of the custom parts. The system is expected to be ready for flight in early 1988. However, continued use of the laboratory system will lead to improvements in the space bioreactor. Those improvements can only be integrated after the initial flight series.

  20. BIOREACTOR LANDFILL DESIGN

    EPA Science Inventory

    Modern landfill design entails many elements including foundations, liner systems, leachate collection systems, stormwater control systems, slope stability considerations, leachate management systems, gas extraction systems, and capping and closure. The use of bioreactor technolo...

  1. Packed Bed Reactor Experiment

    NASA Video Gallery

    The purpose of the Packed Bed Reactor Experiment in low gravity is to determine how a mixture of gas and liquid flows through a packed bed in reduced gravity. A packed bed consists of a metal pipe ...

  2. Anaerobic biodegradation of chloroform under methanogenic conditions

    SciTech Connect

    Devesh, M.T.; Gupta, M.; Suidan, M.T.; Sayles, G.D.

    1994-12-31

    The degradation of chloroform is studied for two different methanogenic cultures grown on acetate and methanol exclusively, as the primary substrates. In chemostats, chloroform was fed at different concentrations as high as 2,000 {micro}g/l along with the primary substrate and chloroform degradation greater than 98% was observed. The kinetics of degradation of chloroform and the primary substrate were investigated using BMP tests and it was seen that the methanol-fed methanogenic culture exhibited higher rates of chloroform degradation than the acetate-fed methanogenic culture. Besides, chloroform inhibited acetate degradation at any concentration while methanol inhibition was observed only for chloroform concentrations higher than 800 {micro}g/l.

  3. Bioreactors and bioseparation.

    PubMed

    Zhang, Siliang; Cao, Xuejun; Chu, Ju; Qian, Jiangchao; Zhuang, Yingping

    2010-01-01

    Along with the rapid development of life science, great attention has been increasingly given to the biotechnological products of cell cultivation technology. In the course of industrialization, bioreactor and bioproduct separation techniques are the two essential technical platforms. In this chapter, the current situation and development prospects of bioreactor techniques in China are systematically discussed, starting with the elucidation of bioreactor processes and the principle of process optimization. Separation technology for biological products is also briefly introduced.At present, a series of bioreactors made by Chinese enterprises have been widely used for laboratory microbial cultivation, process optimization studies, and large-scale production. In the course of bioprocess optimization studies, the complicated bioprocesses in a bioreactor could be resolved into different reaction processes on three scales, namely genetic, cellular, and bioreactor scales. The structural varieties and nonlinear features of various scales of bioprocess systems was discussed through considering the mutual effects of different scale events, namely material flux, energy flux, and information flux, and the optimization approach for bioprocesses was proposed by taking the analysis of metabolic flux and multiscale consideration as a core strategy.In order to realize such an optimization approach, a bioreactor system based on association analysis of multiscale parameters was elaborated, and process optimization of many biological products were materialized, which resulted in great improvement in production efficiency. In designing and manufacturing large-scale bioreactors, the principle of scaling up a process incorporated with flow field study and physiological features in a bioreactor was suggested according to the criterion for the scale-up of cellular physiological and metabolic traits. The flow field features of a bioreactor were investigated through computational fluid

  4. Bioreactors and bioseparation.

    PubMed

    Zhang, Siliang; Cao, Xuejun; Chu, Ju; Qian, Jiangchao; Zhuang, Yingping

    2010-01-01

    Along with the rapid development of life science, great attention has been increasingly given to the biotechnological products of cell cultivation technology. In the course of industrialization, bioreactor and bioproduct separation techniques are the two essential technical platforms. In this chapter, the current situation and development prospects of bioreactor techniques in China are systematically discussed, starting with the elucidation of bioreactor processes and the principle of process optimization. Separation technology for biological products is also briefly introduced.At present, a series of bioreactors made by Chinese enterprises have been widely used for laboratory microbial cultivation, process optimization studies, and large-scale production. In the course of bioprocess optimization studies, the complicated bioprocesses in a bioreactor could be resolved into different reaction processes on three scales, namely genetic, cellular, and bioreactor scales. The structural varieties and nonlinear features of various scales of bioprocess systems was discussed through considering the mutual effects of different scale events, namely material flux, energy flux, and information flux, and the optimization approach for bioprocesses was proposed by taking the analysis of metabolic flux and multiscale consideration as a core strategy.In order to realize such an optimization approach, a bioreactor system based on association analysis of multiscale parameters was elaborated, and process optimization of many biological products were materialized, which resulted in great improvement in production efficiency. In designing and manufacturing large-scale bioreactors, the principle of scaling up a process incorporated with flow field study and physiological features in a bioreactor was suggested according to the criterion for the scale-up of cellular physiological and metabolic traits. The flow field features of a bioreactor were investigated through computational fluid

  5. Isolation and characterization of methanogenic bacteria from landfills

    SciTech Connect

    Fielding, E.R.; Archer, D.B.; De Macario, E.C.; Macario, A.J.L.

    1988-03-01

    Methanogenic bacteria were isolated from landfill sites in the United Kindgom. Strains of Methanobacterium formicicum, Methanosarcina barkeri, several different immunotypes of Methanobacterium bryantii, and a coccoid methanogen distinct from the reference immunotypes were identified.

  6. NASA Classroom Bioreactor

    NASA Technical Reports Server (NTRS)

    Scully, Robert

    2004-01-01

    Exploration of space provides a compelling need for cell-based research into the basic mechanisms that underlie the profound changes that occur in terrestrial life that is transitioned to low gravity environments. Toward that end, NASA developed a rotating bioreactor in which cells are cultured while continuously suspended in a cylinder in which the culture medium rotates with the cylinder. The randomization of the gravity vector accomplished by the continuous rotation, in a low shear environment, provides an analog of microgravity. Because cultures grown in bioreactors develop structures and functions that are much closer to those exhibited by native tissue than can be achieved with traditional culture methods, bioreactors have contributed substantially to advancing research in the fields of cancer, diabetes, infectious disease modeling for vaccine production, drug efficacy, and tissue engineering. NASA has developed a Classroom Bioreactor (CB) that is built from parts that are easily obtained and assembled, user-friendly and versatile. It can be easily used in simple school settings to examine the effect cultures of seeds or cells. An educational brief provides assembly instructions and lesson plans that describes activities in science, math and technology that explore free fall, microgravity, orbits, bioreactors, structure-function relationships and the scientific method.

  7. NASA Bioreactor Schematic

    NASA Technical Reports Server (NTRS)

    2001-01-01

    The schematic depicts the major elements and flow patterns inside the NASA Bioreactor system. Waste and fresh medium are contained in plastic bags placed side-by-side so the waste bag fills as the fresh medium bag is depleted. The compliance vessel contains a bladder to accommodate pressure transients that might damage the system. A peristolic pump moves fluid by squeezing the plastic tubing, thus avoiding potential contamination. The work is sponsored by NASA's Office of Biological and Physical Research. The bioreactor is managed by the Biotechnology Cell Science Program at NASA's Johnson Space Center (JSC). NASA-sponsored bioreactor research has been instrumental in helping scientists to better understand normal and cancerous tissue development. In cooperation with the medical community, the bioreactor design is being used to prepare better models of human colon, prostate, breast and ovarian tumors. Cartilage, bone marrow, heart muscle, skeletal muscle, pancreatic islet cells, liver and kidney are just a few of the normal tissues being cultured in rotating bioreactors by investigators.

  8. Trace Gas Emission from in-Situ Denitrifying Bioreactors

    NASA Astrophysics Data System (ADS)

    Pluer, W.; Walter, M. T.; Geohring, L.

    2014-12-01

    Despite decades of concerted effort to mitigate nonpoint source nitrate (NO3-) pollution from agricultural lands, these efforts have not been sufficient to arrest eutrophication. A primary process for removing excess NO3- from water is denitrification, where denitrifying bacteria use NO3- for respiration in the absence of oxygen. Denitrification results in reduced forms of nitrogen, often dinitrogen gas (N2) but also nitrous oxide (N2O), an aggressive greenhouse gas. A promising solution to NO3- pollution is to intercept agricultural discharges with denitrifying bioreactors (DNBRs). DNBRs provide conditions ideal for denitrifiers: an anaerobic environment, sufficient organic matter, and excess NO3-. These conditions are also ideal for methanogens, which produce methane (CH4), another harmful trace gas. While initial results from bioreactor studies show that they can cost-effectively remove NO3-, trace gas emissions are an unintended consequence. This study's goal was to determine how bioreactor design promotes denitrification while limiting trace gas production. Reactor inflow and outflow water samples were tested for nutrients, including NO3-, and dissolved inflow and outflow gas samples were tested for N2O and CH4. NO3- reduction and trace gas production were evaluated at various residence times, pHs, and inflow NO3- concentrations in field and lab-scale reactors. Low NO3- reduction indicated conditions that stressed denitrifying bacteria while high reductions indicated designs that optimized pollutant treatment for water quality. Several factors influenced high N2O, suggesting non-ideal conditions for the final step of complete denitrification. High CH4 emissions pointed to reactor media choice for discouraging methanogens, which may remove competition with denitrifiers. It is critical to understand all of potential impacts that DNBRs may have, which means identifying processes and design specifications that may affect them.

  9. Development of foamed emulsion bioreactor for air pollution control.

    PubMed

    Kan, Eunsung; Deshusses, Marc A

    2003-10-20

    A new type of bioreactor for air pollution control has been developed. The new process relies on an organic-phase emulsion and actively growing pollutant-degrading microorganisms, made into a foam with the air being treated. This new reactor is referred to as a foamed emulsion bioreactor (FEBR). As there is no packing in the reactor, the FEBR is not subject to clogging. Mathematical modeling of the process and proof of concept using a laboratory prototype revealed that the foamed emulsion bioreactor greatly surpasses the performance of existing gas-phase bioreactors. Experimental results showed a toluene elimination capacity as high as 285 g(toluene) m(-3) (reactor) h(-1) with a removal efficiency of 95% at a gas residence time of 15 s and a toluene inlet concentration of 1-1.3 g x m(-3). Oxygen limited the reactor performance at toluene concentration above about 0.7-1.0 g x m(-3); consequently, performance was significantly improved when pure oxygen was added to the contaminated air. The elimination capacity increased from 204 to 408 g x m(-3) h(-1) with >77% toluene removal at toluene inlet concentrations of 2-2.2 g x m(-3). Overall, the results show that the performance of the FEBR far exceeds that of currently used bioreactors for air pollution control.

  10. Simulation of the inhibition of microbial sulfate reduction in a two-compartment upflow bioreactor subjected to molybdate injection.

    PubMed

    de Jesus, E B; de Andrade Lima, L R P

    2016-08-01

    Souring of oil fields during secondary oil recovery by water injection occurs mainly due to the action of sulfate-reducing bacteria (SRB) adhered to the rock surface in the vicinity of injection wells. Upflow packed-bed bioreactors have been used in petroleum microbiology because of its similarity to the oil field near the injection wells or production. However, these reactors do not realistically describe the regions near the injection wells, which are characterized by the presence of a saturated zone and a void region close to the well. In this study, the hydrodynamics of the two-compartment packing-free/packed-bed pilot bioreactor that mimics an oil reservoir was studied. The packed-free compartment was modeled using a continuous stirred tank model with mass exchange between active and stagnant zones, whereas the packed-bed compartment was modeled using a piston-dispersion-exchange model. The proposed model adequately represents the hydrodynamic of the packed-free/packed-bed bioreactor while the simulations provide important information about the characteristics of the residence time distribution (RTD) curves for different sets of model parameters. Simulations were performed to represent the control of the sulfate-reducing bacteria activity in the bioreactor with the use of molybdate in different scenarios. The simulations show that increased amounts of molybdate cause an effective inhibition of the souring sulfate-reducing bacteria activity. PMID:27126499

  11. NASA Bioreactor tissue culture

    NASA Technical Reports Server (NTRS)

    1998-01-01

    Dr. Lisa E. Freed of the Massachusetts Institute of Technology and her colleagues have reported that initially disc-like specimens tend to become spherical in space, demonstrating that tissues can grow and differentiate into distinct structures in microgravity. The Mir Increment 3 (Sept. 16, 1996 - Jan. 22, 1997) samples were smaller, more spherical, and mechanically weaker than Earth-grown control samples. These results demonstrate the feasibility of microgravity tissue engineering and may have implications for long human space voyages and for treating musculoskeletal disorders on earth. The work is sponsored by NASA's Office of Biological and Physical Research. The bioreactor is managed by the Biotechnology Cell Science Program at NASA's Johnson Space Center (JSC). NASA-sponsored bioreactor research has been instrumental in helping scientists to better understand normal and cancerous tissue development. In cooperation with the medical community, the bioreactor design is being used to prepare better models of human colon, prostate, breast and ovarian tumors. Cartilage, bone marrow, heart muscle, skeletal muscle, pancreatic islet cells, liver and kidney are just a few of the normal tissues being cultured in rotating bioreactors by investigators.

  12. [Acidophilic methanogens and their applications in anaerobic digestion].

    PubMed

    Guo, Xiao-Hui; Wu, Wei-Xiang; Han, Zhi-Ying; Shi, De-Zhi

    2011-02-01

    Methanogens play an important role in global carbon cycle. There exists a range of unknown methanogenic archaea in acidic peat lands, among which, acidophilic methanogens have attracted increasing research interests because of their special metabolic characteristics. To introduce acidophilic methanogens in the anaerobic digestion process of high concentration organic wastes or waste water could essentially overcome the inhibition of acid accumulation on the methanogens and help reduce the operation cost, broadening the industrial application of anaerobic bio-treatment technology. In this paper, we reviewed the recent researches on acidophilic methanogens, with the focus on enrichment and isolation methods, physiological and biochemical characters, metabolic characteristics, and application of molecular biology. The potential applications of acidophilic methanogens in anaerobic digestion process were analyzed and proposed, and the directions for further researches were suggested.

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

    PubMed Central

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

    2015-01-01

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

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

    PubMed

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

    2015-01-01

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

  15. Immobilization of the Methanogenic bacterium methanosarcina barkeri

    SciTech Connect

    Scherer, P.; Kluge, M.; Klein, J.; Sahm, H.

    1981-05-01

    Whole cells of the methanogen Methanosarcina barkeri were immobilized in an alginate network which was crosslinked with Ca/sup 2+/ calcium ions. The rates of methanol conversion to methane of entrapped cells were found to be in the same range as the corresponding rates of free cells. Furthermore, immobilized cells were active for a longer period than free cells. The particle size of the spherical alginate beads and thus diffusion has no obvious influence on the turnover of methanol. The half-value period for methanol conversion activity determined in a buffer medium was approximately 4 days at 37/degree/C for entrapped cells. The high rates of methanol degradation indicated that the immobilization technique preserved the cellular functions of this methanogenic bacterium. 24 refs.

  16. Volatile hydrocarbons inhibit methanogenic crude oil degradation

    PubMed Central

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

    2014-01-01

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

  17. Enzymatic cascade bioreactor

    DOEpatents

    Simmons, Blake A.; Volponi, Joanne V.; Ingersoll, David; Walker, Andrew

    2007-09-04

    Disclosed is an apparatus and method for continuously converting sucrose to .beta.-D-glucose. The method comprises a three stage enzymatic reactor in which an aqueous solution of sucrose is first converted into a solution of fructose and .alpha.-D-glucose by passing it through a porous, packed column containing an inert media on which invertase is immobilized. This solution is then sent through a second packed column containing glucose isomerase and finally a third packed column containing mutarotase. Solution temperature and pH are adjusted to maximize glucose output.

  18. Biotransformation of alkanoylcholines under methanogenic conditions.

    PubMed

    Watson, Mary Katherine; Tezel, Ulas; Pavlostathis, Spyros G

    2012-06-01

    Ester quaternary ammonium compounds (esterquats), which are mainly used as active ingredients in fabric softeners and personal care products, are beginning to replace traditional quaternary ammonium compounds. As a result of hydrophobicity and increasing use, esterquats reach anaerobic treatment systems. However, little is known about the fate of esterquats under anaerobic conditions. In the present study, the potential inhibitory effect and biotransformation of two alkanoylcholines - acetylcholine chloride (ACh-Cl) and lauroylcholine chloride (LCh-Cl) - which are simple esterquats, under methanogenic conditions were investigated. ACh-Cl up to 300 mg/L was not inhibitory to a mixed methanogenic culture. In contrast, methanogenesis was inhibited by LCh-Cl above 50 mg/L, primarily caused by the accumulation of lauric acid which resulted from the abiotic hydrolysis of LCh. Below inhibitory concentrations, both ACh and LCh were transformed to methane by the mixed methanogenic culture. Mass spectrometric analysis confirmed that both alkanoylcholines were first abiotically hydrolyzed to choline and the corresponding alkanoic acid, which were then biotically transformed to methane, carbon dioxide, and ammonia. Thus, alkanoylcholine-containing waste streams can be bioprocessed to form methane, but hydrolysis products such as long-chain alkanoic acids may adversely impact the anaerobic bioconversion of alkanoylcholines. PMID:22463863

  19. Microbiology and biochemistry of the methanogenic archaeobacteria

    NASA Astrophysics Data System (ADS)

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

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

  20. Design and performance of a fibrous bed bioreactor for odor treatment.

    PubMed

    Chua, H; Li, X Z; Yu, P H; Tam, C Y; Huang, Y L; Yang, S T

    2000-01-01

    Biological processes have become popular for odor treatment. In this study, a novel fibrous bed bioreactor was applied for treatment of odorous gas. The column reactor was packed with spirally wound fibrous sheet material on which a consortium of microorganisms selected from activated sludge was immobilized. The first stage of this work comprised a preliminary study that aimed at investigating the feasibility of the fibrous bed bioreactor for treatment of odorous volatile fatty acids (VFAs). In this stage, the performance of a fibrous bed bioreactor at increasing mass loadings ranging from 9.7 to 104.2 g/(m3.h) was studied. VFA removal efficiencies above 90% were achieved at mass loadings up to 50.3 g/(m3.h). At a mass loading of 104.2 g/(m3.h), removal efficiency was found to be 87.7%. In the second stage of the work, the process was scaled up with design and operational considerations, namely, packing medium, process condition, and configuration selections. A trickling biofilter with synthetic fibrous packing medium was selected. It was operated under countercurrent flow of gas and liquid streams. The effects of inlet concentration and empty bed retention time on bioreactor performance were studied. The bioreactor was effective in treating odorous VFAs at mass loadings up to 32 g/(m3.h), at which VFAs started to accumulate in the recirculation liquid, indicating that the biofilm was unable to degrade all the VFAs introduced. Although VFAs accumulated in the liquid phase, the removal efficiency remained above 99%, implying that the biochemical reaction rate, rather than gas-to-liquid mass transfer rate, was the limiting factor of this process. The bioreactor was stable for long-term operation; no clogging and degeneration of the packing medium was observed during the 4-mo operation.

  1. BioReactor

    2003-04-18

    BioReactor is a simulation tool kit for modeling networks of coupled chemical processes (or similar productions rules). The tool kit is implemented in C++ and has the following functionality: 1. Monte Carlo discrete event simulator 2. Solvers for ordinary differential equations 3. Genetic algorithm optimization routines for reverse engineering of models using either Monte Carlo or ODE representation )i.e., 1 or 2)

  2. Decolourisation of textile wastewater in a submerged anaerobic membrane bioreactor.

    PubMed

    Spagni, Alessandro; Casu, Stefania; Grilli, Selene

    2012-08-01

    Azo dye decolourisation can be easily achieved by biological reduction under anaerobic conditions. The aim of this study was to evaluate the applicability of submerged anaerobic membrane bioreactors (SAMBRs) for the decolourisation of dyeing wastewater containing azo dyes. The reactive orange 16 was used as model of an azo dye. The results demonstrated that very high decolourisation (higher than 99%) can be achieved by SAMBRs. Although decolourisation was not significantly influenced by the azo dye concentrations up to 3.2 g L(-1), methane production was greatly inhibited (up to 80-85%). Since volatile fatty acids accumulated in the treatment system with the azo dye concentration increase, methanogenes seem to be the most sensitive microbial populations of the anaerobic ecological community. The results demonstrated that anaerobic process combined with membrane filtration can deal with highly concentrated wastewaters that result from stream separation of industrial discharges.

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

    SciTech Connect

    Leigh, J.A.

    1983-03-01

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

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

    NASA Technical Reports Server (NTRS)

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

    1991-01-01

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

  5. Sulfur formation and recovery in a thiosulfate-oxidizing bioreactor.

    PubMed

    González-Sánchez, A; Meulepas, R; Revah, S

    2008-08-01

    This work describes the design and performance of a thiosulfate-oxidizing bioreactor that allowed high elemental sulfur production and recovery efficiency. The reactor system, referred to as a Supernatant-Recycling Settler Bioreactor (SRSB), consisted of a cylindrical upflow reactor and a separate aeration vessel. The reactor was equipped with an internal settler and packing material (structured corrugated PVC sheets) to facilitate both cell retention and the settling of the formed elemental sulfur. The supernatant from the reactor was continuously recirculated through the aerator. An inlet thiosulfate concentration of 100 mmol l(-1) was used. The reactor system was fed with 89 mmol l(-1) d(-1) thiosulfate reaching 98 to 100% thiosulfate conversion with an elemental sulfur yield of 77%. Ninety-three percent of the produced sulfur was harvested from the bottom of the reactor as sulfur sludge. The dry sulfur sludge contained 87% elemental sulfur. The inclusion of an internal settler and packing material in the reactor system resulted in an effective retention of sulfur and biomass inside the bioreactor, preventing the oxidation of thiosulfate and elemental sulfur to sulfate in the aerator and, therefore, improving the efficiency of elemental sulfur formation and recovery.

  6. Sulfur formation and recovery in a thiosulfate-oxidizing bioreactor.

    PubMed

    González-Sánchez, A; Meulepas, R; Revah, S

    2008-08-01

    This work describes the design and performance of a thiosulfate-oxidizing bioreactor that allowed high elemental sulfur production and recovery efficiency. The reactor system, referred to as a Supernatant-Recycling Settler Bioreactor (SRSB), consisted of a cylindrical upflow reactor and a separate aeration vessel. The reactor was equipped with an internal settler and packing material (structured corrugated PVC sheets) to facilitate both cell retention and the settling of the formed elemental sulfur. The supernatant from the reactor was continuously recirculated through the aerator. An inlet thiosulfate concentration of 100 mmol l(-1) was used. The reactor system was fed with 89 mmol l(-1) d(-1) thiosulfate reaching 98 to 100% thiosulfate conversion with an elemental sulfur yield of 77%. Ninety-three percent of the produced sulfur was harvested from the bottom of the reactor as sulfur sludge. The dry sulfur sludge contained 87% elemental sulfur. The inclusion of an internal settler and packing material in the reactor system resulted in an effective retention of sulfur and biomass inside the bioreactor, preventing the oxidation of thiosulfate and elemental sulfur to sulfate in the aerator and, therefore, improving the efficiency of elemental sulfur formation and recovery. PMID:18724639

  7. High tolerance of methanogens in granular sludge to oxygen

    SciTech Connect

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

    1993-12-01

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

  8. Methanogens: methane producers of the rumen and mitigation strategies.

    PubMed

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

    2010-01-01

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

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

    PubMed

    Shrestha, Pravin Malla; Malvankar, Nikhil S; Werner, Jeffrey J; Franks, Ashley E; Elena-Rotaru, Amelia; Shrestha, Minita; Liu, Fanghua; Nevin, Kelly P; Angenent, Largus T; Lovley, Derek R

    2014-12-01

    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 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 with previous studies, which have demonstrated that Geobacter species can donate electrons to methanogens that are typically predominant in anaerobic digesters, suggest that DIET may be a widespread phenomenon in UASB reactors treating brewery wastes. PMID:25443621

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

    PubMed

    Shrestha, Pravin Malla; Malvankar, Nikhil S; Werner, Jeffrey J; Franks, Ashley E; Elena-Rotaru, Amelia; Shrestha, Minita; Liu, Fanghua; Nevin, Kelly P; Angenent, Largus T; Lovley, Derek R

    2014-12-01

    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 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 with previous studies, which have demonstrated that Geobacter species can donate electrons to methanogens that are typically predominant in anaerobic digesters, suggest that DIET may be a widespread phenomenon in UASB reactors treating brewery wastes.

  11. Oscillating Cell Culture Bioreactor

    NASA Technical Reports Server (NTRS)

    Freed, Lisa E.; Cheng, Mingyu; Moretti, Matteo G.

    2010-01-01

    To better exploit the principles of gas transport and mass transport during the processes of cell seeding of 3D scaffolds and in vitro culture of 3D tissue engineered constructs, the oscillatory cell culture bioreactor provides a flow of cell suspensions and culture media directly through a porous 3D scaffold (during cell seeding) and a 3D construct (during subsequent cultivation) within a highly gas-permeable closed-loop tube. This design is simple, modular, and flexible, and its component parts are easy to assemble and operate, and are inexpensive. Chamber volume can be very low, but can be easily scaled up. This innovation is well suited to work with different biological specimens, particularly with cells having high oxygen requirements and/or shear sensitivity, and different scaffold structures and dimensions. The closed-loop changer is highly gas permeable to allow efficient gas exchange during the cell seeding/culturing process. A porous scaffold, which may be seeded with cells, is fixed by means of a scaffold holder to the chamber wall with scaffold/construct orientation with respect to the chamber determined by the geometry of the scaffold holder. A fluid, with/without biological specimens, is added to the chamber such that all, or most, of the air is displaced (i.e., with or without an enclosed air bubble). Motion is applied to the chamber within a controlled environment (e.g., oscillatory motion within a humidified 37 C incubator). Movement of the chamber induces relative motion of the scaffold/construct with respect to the fluid. In case the fluid is a cell suspension, cells will come into contact with the scaffold and eventually adhere to it. Alternatively, cells can be seeded on scaffolds by gel entrapment prior to bioreactor cultivation. Subsequently, the oscillatory cell culture bioreactor will provide efficient gas exchange (i.e., of oxygen and carbon dioxide, as required for viability of metabolically active cells) and controlled levels of fluid

  12. Detection and quantification of functional genes of cellulose- degrading, fermentative, and sulfate-reducing bacteria and methanogenic archaea.

    PubMed

    Pereyra, L P; Hiibel, S R; Prieto Riquelme, M V; Reardon, K F; Pruden, A

    2010-04-01

    Cellulose degradation, fermentation, sulfate reduction, and methanogenesis are microbial processes that coexist in a variety of natural and engineered anaerobic environments. Compared to the study of 16S rRNA genes, the study of the genes encoding the enzymes responsible for these phylogenetically diverse functions is advantageous because it provides direct functional information. However, no methods are available for the broad quantification of these genes from uncultured microbes characteristic of complex environments. In this study, consensus degenerate hybrid oligonucleotide primers were designed and validated to amplify both sequenced and unsequenced glycoside hydrolase genes of cellulose-degrading bacteria, hydA genes of fermentative bacteria, dsrA genes of sulfate-reducing bacteria, and mcrA genes of methanogenic archaea. Specificity was verified in silico and by cloning and sequencing of PCR products obtained from an environmental sample characterized by the target functions. The primer pairs were further adapted to quantitative PCR (Q-PCR), and the method was demonstrated on samples obtained from two sulfate-reducing bioreactors treating mine drainage, one lignocellulose based and the other ethanol fed. As expected, the Q-PCR analysis revealed that the lignocellulose-based bioreactor contained higher numbers of cellulose degraders, fermenters, and methanogens, while the ethanol-fed bioreactor was enriched in sulfate reducers. The suite of primers developed represents a significant advance over prior work, which, for the most part, has targeted only pure cultures or has suffered from low specificity. Furthermore, ensuring the suitability of the primers for Q-PCR provided broad quantitative access to genes that drive critical anaerobic catalytic processes.

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

    PubMed

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

    2014-12-01

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

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

    PubMed Central

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

    2014-01-01

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

  15. Progress in the development of vaccines against rumen methanogens.

    PubMed

    Wedlock, D N; Janssen, P H; Leahy, S C; Shu, D; Buddle, B M

    2013-06-01

    Vaccination against rumen methanogens offers a practical approach to reduce methane emissions in livestock, particularly ruminants grazing on pasture. Although successful vaccination strategies have been reported for reducing the activity of the rumen-dwelling organism Streptococcus bovis in sheep and S. bovis and Lactobacillus spp. in cattle, earlier approaches using vaccines based on whole methanogen cells to reduce methane production in sheep have produced less promising results. An anti-methanogen vaccine will need to have broad specificity against methanogens commonly found in the rumen and induce antibody in saliva resulting in delivery of sufficiently high levels of antibodies to the rumen to reduce methanogen activity. Our approach has focussed on identifying surface and membrane-associated proteins that are conserved across a range of rumen methanogens. The identification of potential vaccine antigens has been assisted by recent advances in the knowledge of rumen methanogen genomes. Methanogen surface proteins have been shown to be immunogenic in ruminants and vaccination of sheep with these proteins induced specific antibody responses in saliva and rumen contents. Current studies are directed towards identifying key candidate antigens and investigating the level and types of salivary antibodies produced in sheep and cattle vaccinated with methanogen proteins, stability of antibodies in the rumen and their impact on rumen microbial populations. In addition, there is a need to identify adjuvants that stimulate high levels of salivary antibody and are suitable for formulating with protein antigens to produce a low-cost and effective vaccine.

  16. Design challenges for space bioreactors

    NASA Technical Reports Server (NTRS)

    Seshan, P. K.; Petersen, G. R.

    1989-01-01

    The design of bioreactors for operation under conditions of microgravity presents problems and challenges. Absence of a significant body force such as gravity can have profound consequences for interfacial phenomena. Marangoni convection can no longer be overlooked. Many speculations on the advantages and benefits of microgravity can be found in the literature. Initial bioreactor research considerations for space applications had little regard for the suitability of the designs for conditions of microgravity. Bioreactors can be classified in terms of their function and type of operation. The complex interaction of parameters leading to optimal design and operation of a bioreactor is illustrated by the JSC mammalian cell culture system. The design of a bioreactor is strongly dependent upon its intended use as a production unit for cell mass and/or biologicals or as a research reactor for the study of cell growth and function. Therefore a variety of bioreactor configurations are presented in rapid summary. Following this, a rationale is presented for not attempting to derive key design parameters such as the oxygen transfer coefficient from ground-based data. A set of themes/objectives for flight experiments to develop the expertise for design of space bioreactors is then proposed for discussion. These experiments, carried out systematically, will provide a database from which engineering tools for space bioreactor design will be derived.

  17. Characteristics, Process Parameters, and Inner Components of Anaerobic Bioreactors

    PubMed Central

    Abdelgadir, Awad; Chen, Xiaoguang; Liu, Jianshe; Xie, Xuehui; Zhang, Jian; Zhang, Kai; Wang, Heng; Liu, Na

    2014-01-01

    The anaerobic bioreactor applies the principles of biotechnology and microbiology, and nowadays it has been used widely in the wastewater treatment plants due to their high efficiency, low energy use, and green energy generation. Advantages and disadvantages of anaerobic process were shown, and three main characteristics of anaerobic bioreactor (AB), namely, inhomogeneous system, time instability, and space instability were also discussed in this work. For high efficiency of wastewater treatment, the process parameters of anaerobic digestion, such as temperature, pH, Hydraulic retention time (HRT), Organic Loading Rate (OLR), and sludge retention time (SRT) were introduced to take into account the optimum conditions for living, growth, and multiplication of bacteria. The inner components, which can improve SRT, and even enhance mass transfer, were also explained and have been divided into transverse inner components, longitudinal inner components, and biofilm-packing material. At last, the newly developed special inner components were discussed and found more efficient and productive. PMID:24672798

  18. Redox zones stratification and the microbial community characteristics in a periphyton bioreactor.

    PubMed

    Liu, Junzhuo; Liu, Wei; Wang, Fengwu; Kerr, Philip; Wu, Yonghong

    2016-03-01

    Bioremediation techniques based on microorganisms have been widely applied to treat polluted surface water, but the efficiencies have been limited, especially in deep and static waters. Microbial aggregates, known as periphyton, were introduced into a tank bioreactor to improve pollutants removal and a periphyton bioreactor with an 84 cm column was built to investigate microbe-wastewater interactions. Periphyton greatly improved water quality and produced a distinct stratification in the water column into five redox zones with slight overlaps. From top to bottom these were: oxygen reduction, nitrate reduction, iron reduction, sulfate reduction and methanogenic zone. Periphyton communities had high species diversities (767-947 OTUs) with the facultative zone (middle layer) having higher species richness and functional diversity than the aerobic (top layer) and anaerobic zones (bottom layer). A good knowledge of interactions between periphyton and water column stratification could benefit from integration of periphyton to improve bioremediation of deep and static water.

  19. Evaluation of molecular methods used for establishing the interactions and functions of microorganisms in anaerobic bioreactors.

    PubMed

    Talbot, G; Topp, E; Palin, M F; Massé, D I

    2008-02-01

    Molecular techniques have unveiled the complexity of the microbial consortium in anaerobic bioreactors and revealed the presence of several uncultivated species. This paper presents a review of the panoply of classical and recent molecular approaches and multivariate analyses that have been, or might be used to establish the interactions and functions of these anaerobic microorganisms. Most of the molecular approaches used so far are based on the analysis of small subunit ribosomal RNA but recent studies also use quantification of functional gene expressions. There are now several studies that have developed quantitative real-time PCR assays to investigate methanogens. With a view to improving the stability and performance of bioreactors, monitoring with molecular methods is also discussed. Advances in metagenomics and proteomics will lead to the development of promising lab-on chip technologies for cost-effective monitoring.

  20. Perfusion Bioreactor Module

    NASA Technical Reports Server (NTRS)

    Morrison, Dennis R.

    1990-01-01

    Perfusion bioreactor module, self-contained, closed-loop cell-culture system that operates in microgravity or on Earth. Equipment supports growth or long-term maintenance of cultures of human or other fragile cells for experiments in basic cell biology or process technology. Designed to support proliferation (initially at exponential rates of growth) of cells in complex growth medium and to maintain confluent cells in defined medium under conditions optimized to permit or encourage selected functions of cells, including secretion of products of cells into medium.

  1. Optimal packings of superballs

    NASA Astrophysics Data System (ADS)

    Jiao, Y.; Stillinger, F. H.; Torquato, S.

    2009-04-01

    Dense hard-particle packings are intimately related to the structure of low-temperature phases of matter and are useful models of heterogeneous materials and granular media. Most studies of the densest packings in three dimensions have considered spherical shapes, and it is only more recently that nonspherical shapes (e.g., ellipsoids) have been investigated. Superballs (whose shapes are defined by |x1|2p+|x2|2p+|x3|2p≤1 ) provide a versatile family of convex particles (p≥0.5) with both cubic-like and octahedral-like shapes as well as concave particles (0packings for all convex and concave cases. The candidate maximally dense packings are certain families of Bravais lattice packings (in which each particle has 12 contacting neighbors) possessing the global symmetries that are consistent with certain symmetries of a superball. We also provide strong evidence that our packings for convex superballs (p≥0.5) are most likely the optimal ones. The maximal packing density as a function of p is nonanalytic at the sphere point (p=1) and increases dramatically as p moves away from unity. Two more nontrivial nonanalytic behaviors occur at pc∗=1.1509… and po∗=ln3/ln4=0.7924… for “cubic” and “octahedral” superballs, respectively, where different Bravais lattice packings possess the same densities. The packing characteristics determined by the broken rotational symmetry of superballs are similar to but richer than their two-dimensional “superdisk” counterparts [Y. Jiao , Phys. Rev. Lett. 100, 245504 (2008)] and are distinctly different from that of ellipsoid packings. Our candidate optimal superball packings provide a starting point to quantify the equilibrium phase behavior of superball systems, which should deepen our understanding of the statistical thermodynamics of nonspherical-particle systems.

  2. Pulse shear stress for anaerobic membrane bioreactor fouling control.

    PubMed

    Yang, Jixiang; Spanjers, Henri; van Lier, Jules B

    2011-01-01

    Increase of shear stress at membrane surfaces is a generally applied strategy to minimize membrane fouling. It has been reported that a two-phase flow, better known as slug flow, is an effective way to increase shear stress. Hence, slug flow was introduced into an anaerobic membrane bioreactor for membrane fouling control. Anaerobic suspended sludge was cultured in an anaerobic membrane bioreactor (AMBR) operated with a side stream inside-out tubular membrane unit applying sustainable flux flow regimes. The averaged particle diameter decreased from 20 to 5 microm during operation of the AMBR. However, the COD removal efficiency did not show any significant deterioration, whereas the specific methanogenic activity (SMA) increased from 0.16 to 0.41 gCOD/g VSS/day. Nevertheless, the imposed gas slug appeared to be insufficient for adequate fouling control, resulting in rapidly increasing trans membrane pressures (TMP) operating at a flux exceeding 16 L/m2/h. Addition of powdered activated carbon (PAC) enhanced the effect of slug flow on membrane fouling. However, the combined effect was still considered as not being significant. The tubular membrane was subsequently equipped with inert inserts for creating a locally increased shear stress for enhanced fouling control. Results show an increase in the membrane flux from 16 L/m2/h to 34 L/m2/h after the inserts were mounted in the membrane tube. PMID:22097007

  3. Genomic Characterization of Methanomicrobiales Reveals Three Classes of Methanogens

    SciTech Connect

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

    2009-01-01

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

  4. Genomic Characterization of Methanomicrobiales Reveals Three Classes of Methanogens

    SciTech Connect

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

    2009-05-01

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

  5. Microtechnology in space bioreactors.

    PubMed

    Walther, I; van der Schoot, B; Boillat, M; Muller, O; Cogoli, A

    1999-03-01

    Space biology is a young and rapidly developing discipline comprising basic research and biotechnology. In the next decades it will play a prominent role in the International Space Station (ISS). Therefore, there is an increasing demand for sophisticated instrumentation to satisfy the requirements of the future projects in space biology. Bioreactors will be needed to supply fresh living material (cells and tissues) either to study still obscure basic biological mechanisms or to develop profitable bioprocesses which will take advantage of the peculiar microgravity conditions. Since more than twenty years, the Space Biology Group of the ETHZ is carrying out research projects in space (Space Shuttle/Spacelab, MIR Station, satellites, and sounding rockets) that involve also the development of space-qualified instrumentation. In the last ten years we have developed, in collaboration with Mecanex SA, Nyon, and the Institute of Microtechnology of the University of Neuchatel, a space bioreactor for the continuous culture of yeast cells under controlled conditions. Sensors, pH control, nutrients pump and fluid flowmeter are based on state-of-the-art silicon technology. After two successful space flights, a further improved version is presently prepared for a flight in the year 2000.

  6. Sensing in tissue bioreactors

    NASA Astrophysics Data System (ADS)

    Rolfe, P.

    2006-03-01

    Specialized sensing and measurement instruments are under development to aid the controlled culture of cells in bioreactors for the fabrication of biological tissues. Precisely defined physical and chemical conditions are needed for the correct culture of the many cell-tissue types now being studied, including chondrocytes (cartilage), vascular endothelial cells and smooth muscle cells (blood vessels), fibroblasts, hepatocytes (liver) and receptor neurones. Cell and tissue culture processes are dynamic and therefore, optimal control requires monitoring of the key process variables. Chemical and physical sensing is approached in this paper with the aim of enabling automatic optimal control, based on classical cell growth models, to be achieved. Non-invasive sensing is performed via the bioreactor wall, invasive sensing with probes placed inside the cell culture chamber and indirect monitoring using analysis within a shunt or a sampling chamber. Electroanalytical and photonics-based systems are described. Chemical sensing for gases, ions, metabolites, certain hormones and proteins, is under development. Spectroscopic analysis of the culture medium is used for measurement of glucose and for proteins that are markers of cell biosynthetic behaviour. Optical interrogation of cells and tissues is also investigated for structural analysis based on scatter.

  7. Flat Pack Toy Design

    ERIC Educational Resources Information Center

    Hutcheson, Brian

    2007-01-01

    In this article, the author introduces the concept of flat pack toys. Flat pack toys are designed using a template on a single sheet of letter-sized card stock paper. Before being cut out and built into a three-dimensional toy, they are scanned into the computer and uploaded to a website. With the template accessible from the website, anyone with…

  8. TLC Pack Unpacked

    ERIC Educational Resources Information Center

    Oberhofer, Margret; Colpaert, Jozef

    2015-01-01

    TLC Pack stands for Teaching Languages to Caregivers and is a course designed to support migrants working or hoping to work in the caregiving sector. The TLC Pack resources range from A2 to B2 level of the Common European Framework of Reference for Languages (CEFR), and will be made available online in the six project languages: Dutch, English,…

  9. Cells growing in NASA Bioreactor

    NASA Technical Reports Server (NTRS)

    1998-01-01

    For 5 days on the STS-70 mission, a bioreactor cultivated human colon cancer cells, which grew to 30 times the volume of control specimens grown on Earth. This significant result was reproduced on STS-85 which grew mature structures that more closely match what are found in tumors in humans. Shown here, clusters of cells slowly spin inside a bioreactor. On Earth, the cells continually fall through the buffer medium and never hit bottom. In space, they are naturally suspended. Rotation ensures gentle stirring so waste is removed and fresh nutrient and oxygen are supplied. The NASA Bioreactor provides a low turbulence culture environment which promotes the formation of large, three-dimensional cell clusters. Due to their high level of cellular organization and specialization, samples constructed in the bioreactor more closely resemble the original tumor or tissue found in the body. The Bioreactor is rotated to provide gentle mixing of fresh and spent nutrient without inducing shear forces that would damage the cells. The work is sponsored by NASA's Office of Biological and Physical Research. The bioreactor is managed by the Biotechnology Cell Science Program at NASA's Johnson Space Center (JSC). NASA-sponsored bioreactor research has been instrumental in helping scientists to better understand normal and cancerous tissue development. In cooperation with the medical community, the bioreactor design is being used to prepare better models of human colon, prostate, breast and ovarian tumors. Cartilage, bone marrow, heart muscle, skeletal muscle, pancreatic islet cells, liver and kidney are just a few of the normal tissues being cultured in rotating bioreactors by investigators.

  10. Energetics of syntrophic cooperation in methanogenic degradation.

    PubMed Central

    Schink, B

    1997-01-01

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

  11. Graphitic packing removal tool

    SciTech Connect

    Meyers, K.E.; Kolsun, G.J.

    1996-12-31

    Graphitic packing removal tools are described for removal of the seal rings in one piece from valves and pumps. The packing removal tool has a cylindrical base ring the same size as the packing ring with a surface finish, perforations, knurling or threads for adhesion to the seal ring. Elongated leg shanks are mounted axially along the circumferential center. A slit or slits permit insertion around shafts. A removal tool follower stabilizes the upper portion of the legs to allow a spanner wrench to be used for insertion and removal.

  12. Graphitic packing removal tool

    DOEpatents

    Meyers, K.E.; Kolsun, G.J.

    1997-11-11

    Graphitic packing removal tools for removal of the seal rings in one piece are disclosed. The packing removal tool has a cylindrical base ring the same size as the packing ring with a surface finish, perforations, knurling or threads for adhesion to the seal ring. Elongated leg shanks are mounted axially along the circumferential center. A slit or slits permit insertion around shafts. A removal tool follower stabilizes the upper portion of the legs to allow a spanner wrench to be used for insertion and removal. 5 figs.

  13. Graphitic packing removal tool

    DOEpatents

    Meyers, Kurt Edward; Kolsun, George J.

    1997-01-01

    Graphitic packing removal tools for removal of the seal rings in one piece. he packing removal tool has a cylindrical base ring the same size as the packing ring with a surface finish, perforations, knurling or threads for adhesion to the seal ring. Elongated leg shanks are mounted axially along the circumferential center. A slit or slits permit insertion around shafts. A removal tool follower stabilizes the upper portion of the legs to allow a spanner wrench to be used for insertion and removal.

  14. Evaluation of different configurations of hybrid membrane bioreactors for treatment of domestic wastewater.

    PubMed

    Cuevas-Rodríguez, G; Cervantes-Avilés, P; Torres-Chávez, I; Bernal-Martínez, A

    2015-01-01

    Four membrane bioreactors (MBRs) with the same dimensions were studied for 180 days: three hybrid growth membrane bioreactors with biofilm attached in different packing media and a conventional MBR (C-MBR). The four MBRs had an identical membrane module of hollow fiber with a nominal porous diameter of 0.4 μm. The MBRs were: (1) a C-MBR; (2) a moving bed membrane bioreactor (MB-MBR), which was packed with 2 L of carrier Kaldnes-K1, presenting an exposed surface area of 678.90 m²/m³; (3) a non-submerged organic fixed bed (OFB-MBR) packed with 6.5 L of organic packing media composed of a mixture of cylindrical pieces of wood, providing an exposed surface area of 178.05 m²/m³; and (4) an inorganic fixed bed non-submerged membrane bioreactor (IFB-MBR) packed with 6 L of spherical volcanic pumice stone with an exposed surface area of 526.80 m²/m³. The four MBRs were fed at low organic loading (0.51 ± 0.19 kgCOD/m³ d). The results were recorded according to the behavior of the total resistance, transmembrane pressure (TMP), permeability, and removal percentages of the nutrients during the experimental time. The results showed that the MB-MBR presented the better performance on membrane filtration, while the higher nutrient removals were detected in the OFB-MBR and IFB-MBR.

  15. Evaluation of different configurations of hybrid membrane bioreactors for treatment of domestic wastewater.

    PubMed

    Cuevas-Rodríguez, G; Cervantes-Avilés, P; Torres-Chávez, I; Bernal-Martínez, A

    2015-01-01

    Four membrane bioreactors (MBRs) with the same dimensions were studied for 180 days: three hybrid growth membrane bioreactors with biofilm attached in different packing media and a conventional MBR (C-MBR). The four MBRs had an identical membrane module of hollow fiber with a nominal porous diameter of 0.4 μm. The MBRs were: (1) a C-MBR; (2) a moving bed membrane bioreactor (MB-MBR), which was packed with 2 L of carrier Kaldnes-K1, presenting an exposed surface area of 678.90 m²/m³; (3) a non-submerged organic fixed bed (OFB-MBR) packed with 6.5 L of organic packing media composed of a mixture of cylindrical pieces of wood, providing an exposed surface area of 178.05 m²/m³; and (4) an inorganic fixed bed non-submerged membrane bioreactor (IFB-MBR) packed with 6 L of spherical volcanic pumice stone with an exposed surface area of 526.80 m²/m³. The four MBRs were fed at low organic loading (0.51 ± 0.19 kgCOD/m³ d). The results were recorded according to the behavior of the total resistance, transmembrane pressure (TMP), permeability, and removal percentages of the nutrients during the experimental time. The results showed that the MB-MBR presented the better performance on membrane filtration, while the higher nutrient removals were detected in the OFB-MBR and IFB-MBR. PMID:25714631

  16. Spiral vane bioreactor

    NASA Technical Reports Server (NTRS)

    Morrison, Dennis R. (Inventor)

    1991-01-01

    A spiral vane bioreactor of a perfusion type is described in which a vertical chamber, intended for use in a microgravity condition, has a central rotating filter assembly and has flexible membranes disposed to rotate annularly about the filter assembly. The flexible members have end portions disposed angularly with respect to one another. A fluid replenishment medium is input from a closed loop liquid system to a completely liquid filled chamber containing microcarrier beads, cells and a fluid medium. Output of spent medium is to the closed loop. In the closed loop, the output and input parameters are sensed by sensors. A manifold permits recharging of the nutrients and pH adjustment. Oxygen is supplied and carbon dioxide and bubbles are removed and the system is monitored and controlled by a microprocessor.

  17. Controlled-Turbulence Bioreactors

    NASA Technical Reports Server (NTRS)

    Wolf, David A.; Schwartz, Ray; Trinh, Tinh

    1989-01-01

    Two versions of bioreactor vessel provide steady supplies of oxygen and nutrients with little turbulence. Suspends cells in environment needed for sustenance and growth, while inflicting less damage from agitation and bubbling than do propeller-stirred reactors. Gentle environments in new reactors well suited to delicate mammalian cells. One reactor kept human kidney cells alive for as long as 11 days. Cells grow on carrier beads suspended in liquid culture medium that fills cylindrical housing. Rotating vanes - inside vessel but outside filter - gently circulates nutrient medium. Vessel stationary; magnetic clutch drives filter cylinder and vanes. Another reactor creates even less turbulence. Oxygen-permeable tubing wrapped around rod extending along central axis. Small external pump feeds oxygen to tubing through rotary coupling, and oxygen diffuses into liquid medium.

  18. Microbial Bioreactor Development in the ALS NSCORT

    NASA Astrophysics Data System (ADS)

    Mitchell, Cary; Whitaker, Dawn; Banks, M. Katherine; Heber, Albert J.; Turco, Ronald F.; Nies, Loring F.; Alleman, James E.; Sharvelle, Sybil E.; Li, Congna; Heller, Megan

    and recycling of effluent supernatant were evaluated to maximize degradation and minimize water input. The off-gases proceeded to a bioregenerative air-treatment reactor, and the sludge effluent was investigated for multiple downstream uses including dewatering by reed beds, use as a nutrient supplement for fish or mushroom growth, and as a growth medium and nutrient source for various crops. The Bio-Regenerative Environmental Air Treatment for Health (BREATHe I) reactor treated greywater and off-gases from the thermophilic aerobic digestion reactor which contained elevated levels of ammonia (NH3 ) and hydrogen sulfide (H2 S). BREATHe I development focused initially on removing greywater contaminants with clean air supplied to a biotrickling filter. Limited removal of organic carbon (70%) led to studies indicating that biodegradation metabolites of the surfactant disodium cocoamphodiacetate are recalcitrant. Subsequent studies showed that NH3 loaded at 150 mg/min and H2 S at 0.83 mg/min were removed completely, while removal of carbonaceous compounds from greywater remained constant. A BREATHe II reactor emphasized biofilters and biotrickling filters for removal of ersatz multicomponent gaseous waste streams representative of habitat air and atmospheric condensate. The model waste stream contained a mixture of acetone, n-butanol, methane, ethylene, and ammonia. Both biofilters and biotrickling filters packed with different media were able to achieve complete removal of easily soluble compounds such as acetone, n-butanol, and ammonia within a short startup period, whereas methane was not removed because of its extreme aqueous insolubility. Different packing media and bioreactor configurations were subsequently assessed, as well as the effect of influent ammonia concentration. Research sponsored in part by NASA grant NAG5-12686.

  19. Membrane Bioreactor With Pressure Cycle

    NASA Technical Reports Server (NTRS)

    Efthymiou, George S.; Shuler, Michael L.

    1991-01-01

    Improved class of multilayer membrane bioreactors uses convention forced by differences in pressure to overcome some of diffusional limitations of prior bioreactors. In reactor of new class, flow of nutrient solution reduces adverse gradients of concentration, keeps cells supplied with fresh nutrient, and sweeps away products faster than diffusion alone. As result, overall yield and rate of reaction increased. Pressures in sweeping gas and nutrient alternated to force nutrient liquid into and out of biocatalyst layer through hyrophilic membrane.

  20. Bioreactor Mass Transport Studies

    NASA Technical Reports Server (NTRS)

    Kleis, Stanley J.; Begley, Cynthia M.

    1997-01-01

    The objectives of the proposed research efforts were to develop both a simulation tool and a series of experiments to provide a quantitative assessment of mass transport in the NASA rotating wall perfused vessel (RWPV) bioreactor to be flown on EDU#2. This effort consisted of a literature review of bioreactor mass transport studies, the extension of an existing scalar transport computer simulation to include production and utilization of the scalar, and the evaluation of experimental techniques for determining mass transport in these vessels. Since mass transport at the cell surface is determined primarily by the relative motion of the cell assemblage and the surrounding fluid, a detailed assessment of the relative motion was conducted. Results of the simulations of the motion of spheres in the RWPV under microgravity conditions are compared with flight data from EDU#1 flown on STS-70. The mass transport across the cell membrane depends upon the environment, the cell type, and the biological state of the cell. Results from a literature review of cell requirements of several scalars are presented. As a first approximation, a model with a uniform spatial distribution of utilization or production was developed and results from these simulations are presented. There were two candidate processes considered for the experimental mass transport evaluations. The first was to measure the dissolution rate of solid or gel beads. The second was to measure the induced fluorescence of beads as a stimulant (for example hydrogen peroxide) is infused into the vessel. Either technique would use video taped images of the process for recording the quantitative results. Results of preliminary tests of these techniques are discussed.

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

    PubMed

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

    2015-09-01

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

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

    PubMed

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

    2015-09-01

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

  3. Influent wastewater microbiota and temperature influence anaerobic membrane bioreactor microbial community.

    PubMed

    Seib, M D; Berg, K J; Zitomer, D H

    2016-09-01

    Sustainable municipal wastewater recovery scenarios highlight benefits of anaerobic membrane bioreactors (AnMBRs). However, influences of continuous seeding by influent wastewater and temperature on attached-growth AnMBRs are not well understood. In this study, four bench-scale AnMBR operated at 10 and 25°C were fed synthetic (SPE) and then real (PE) primary effluent municipal wastewater. Illumina sequencing revealed different bacterial communities in each AnMBR in response to temperature and bioreactor configuration, whereas differences were not observed in archaeal communities. Activity assays revealed hydrogenotrophic methanogenesis was the dominant methanogenic pathway at 10°C. The significant relative abundance of Methanosaeta at 10°C concomitant with low acetoclastic methanogenic activity may indicate possible Methanosaeta-Geobacter direct interspecies electron transfer. When AnMBR feed was changed to PE, continual seeding with wastewater microbiota caused AnMBR microbial communities to shift, becoming more similar to PE microbiota. Therefore, influent wastewater microbiota, temperature and reactor configuration influenced the AnMBR microbial community. PMID:27262719

  4. Nasal packing and stenting

    PubMed Central

    Weber, Rainer K.

    2011-01-01

    Nasal packs are indispensable in ENT practice. This study reviews current indications, effectiveness and risks of nasal packs and stents. In endoscopic surgery, nasal packs should always have smooth surfaces to minimize mucosal damage, improve wound healing and increase patient comfort. Functional endoscopic endonasal sinus surgery allows the use of modern nasal packs, since pressure is no longer required. So called hemostatic/resorbable materials are a first step in this direction. However, they may lead to adhesions and foreign body reactions in mucosal membranes. Simple occlusion is an effective method for creating a moist milieu for improved wound healing and avoiding dryness. Stenting of the frontal sinus is recommended if surgery fails to produce a wide, physiologically shaped drainage path that is sufficiently covered by intact tissue. PMID:22073095

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

    PubMed

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

    2012-08-21

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

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

    PubMed

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

    2012-08-21

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

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

    PubMed Central

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

    2012-01-01

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

  8. The potential of Methanogenic Life in the Solar System

    NASA Astrophysics Data System (ADS)

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

    2015-10-01

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

  9. Methanogenic Conversion of CO2 Into CH4

    SciTech Connect

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

    2012-05-06

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

  10. Acidification of methanol-fed anaerobic granular sludge bioreactors by cobalt deprivation: Induction and microbial community dynamics.

    PubMed

    Fermoso, Fernando G; Collins, Gavin; Bartacek, Jan; O'Flaherty, Vincent; Lens, Piet

    2008-01-01

    The acidification of mesophilic (30 degrees C) methanol-fed upflow anaerobic sludge bed (UASB) reactors induced by cobalt deprivation from the influent was investigated by coupling the reactor performance (pH 7.0; organic loading rate 4.5 g COD . L(-1) . d(-1)) to the microbial ecology of the bioreactor sludge. The latter was investigated by specific methanogenic activity (SMA) measurements and fluorescence in situ hybridization (FISH) to quantify the abundance of key organisms over time. This study hypothesized that under cobalt limiting conditions, the SMA on methanol of the sludge gradually decreases, which ultimately results in methanol accumulation in the reactor effluent. Once the methanol accumulation surpasses a threshold value (about 8.5 mM for the sludge investigated), reactor acidification occurs because acetogens outcompete methylothrophic methanogens at these elevated methanol concentrations. Methanogens present in granular sludge at the time of the acidification do not use methanol as the direct substrate and are unable to degrade acetate. Methylotrophic/acetoclastic methanogenic activity was found to be lost within 10 days of reactor operation, coinciding with the disappearance of the Methanosarcina population. The loss of SMA on methanol can thus be used as an accurate parameter to predict reactor acidification of methanol-fed UASB reactors operating under cobalt limiting conditions.

  11. Supermacroporous polymer-based cryogel bioreactor for monoclonal antibody production in continuous culture using hybridoma cells.

    PubMed

    Jain, Era; Karande, Anjali A; Kumar, Ashok

    2011-01-01

    Cryogel matrices composed of different polymeric blends were synthesized, yielding a unique combination of hydrophilicity and hydrophobicity with the presence or absence of charged surface. Four such cryogel matrices composed of polyacrylamide-chitosan (PAAC), poly(N-isopropylacrylamide)-chitosan, polyacrylonitrile (PAN), and poly(N-isopropylacrylamide) were tested for growth of different hybridoma cell lines and production of antibody in static culture. All the matrices were capable for the adherence of hybridoma cell lines 6A4D7, B7B10, and H9E10 to the polymeric surfaces as well as for the efficient monoclonal antibody (mAb) production. PAAC proved to be relatively better in terms of both mAb production and cell growth. Further, PAAC cryogel was designed into three different formats, monolith, disks, and beads, and used as packing material for packed-bed bioreactor. Long-term cultivation of 6A4D7 cell line on PAAC cryogel scaffold in all the three formats could be successfully done for a period of 6 weeks under static conditions. Continuous packed-bed bioreactor was setup using 6A4D7 hybridoma cell line in the three reactor formats. The reactors ran continuously for a period of 60 days during which mAb production and metabolism of cells in the bioreactors were monitored periodically. The monolith bioreactor performed most efficiently over a period of 60 days and produced a total of 57.5 mg of antibody in the first 30 days (in 500 mL) with a highest concentration of 115 μg mL(-1) , which is fourfold higher than t-flask culture. The results demonstrate that appropriate chemistry and geometry of the bioreactor matrix for cell growth and immobilization can enhance the reactor productivity. PMID:20865749

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

    USGS Publications Warehouse

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

    2002-01-01

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

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

    PubMed

    Chapelle, Francis H; O'Neill, Kathleen; Bradley, Paul M; Methé, Barbara A; Ciufo, Stacy A; Knobel, LeRoy L; Lovley, Derek R

    2002-01-17

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

  14. A comment on methanogenic bacteria and the primitive ecology

    NASA Technical Reports Server (NTRS)

    Woese, C. R.

    1977-01-01

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

  15. Space bioreactor: Design/process flow

    NASA Technical Reports Server (NTRS)

    Cross, John H.

    1987-01-01

    The design of the space bioreactor stems from three considerations. First, and foremost, it must sustain cells in microgravity. Closely related is the ability to take advantage of the weightlessness and microgravity. Lastly, it should fit into a bioprocess. The design of the space bioreactor is described in view of these considerations. A flow chart of the bioreactor is presented and discussed.

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

    PubMed

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

    2016-05-01

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

  17. Methanogenic archaea database containing physiological and biochemical characteristics.

    PubMed

    Jabłoński, Sławomir; Rodowicz, Paweł; Łukaszewicz, Marcin

    2015-04-01

    The methanogenic archaea are a group of micro-organisms that have developed a unique metabolic pathway for obtaining energy. There are 150 characterized species in this group; however, novel species continue to be discovered. Since methanogens are considered a crucial part of the carbon cycle in the anaerobic ecosystem, characterization of these micro-organisms is important for understanding anaerobic ecology. A methanogens database (MDB; http://metanogen.biotech.uni.wroc.pl/), including physiological and biochemical characteristics of methanogens, was constructed based on the descriptions of isolated type strains. Analysis of the data revealed that methanogens are able to grow from 0 to 122 °C. Methanogens growing at the same temperature may have very different growth rates. There is no clear correlation between the optimal growth temperature and the DNA G+C content. The following substrate preferences are observed in the database: 74.5% of archaea species utilize H2+CO2, 33% utilize methyl compounds and 8.5% utilize acetate. Utilization of methyl compounds (mainly micro-organisms belonging to the genera Methanosarcina and Methanolobus ) is seldom accompanied by an ability to utilize H2+CO2. Very often, data for described species are incomplete, especially substrate preferences. Additional research leading to completion of missing information and development of standards, especially for substrate utilization, would be very helpful.

  18. Evidence for para dechlorination of polychlorobiphenyls by methanogenic bacteria

    SciTech Connect

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

    1995-06-01

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

  19. Anaerobic digestion of secondary residuals from an anaerobic bioreactor at a brewery to enhance bioenergy generation.

    PubMed

    Bocher, Benjamin T; Agler, Matthew T; Garcia, Marcelo L; Beers, Allen R; Angenent, Largus T

    2008-05-01

    Many beer breweries use high-rate anaerobic digestion (AD) systems to treat their soluble high-strength wastewater. Biogas from these AD systems is used to offset nonrenewable energy utilization in the brewery. With increasing nonrenewable energy costs, interest has mounted to also digest secondary residuals from the high-rate digester effluent, which consists of yeast cells, bacteria, methanogens, and small (hemi)cellulosic particles. Mesophilic (37 degrees C) and thermophilic (55 degrees C) lab-scale, low-rate continuously-stirred anaerobic digestion (CSAD) bioreactors were operated for 258 days by feeding secondary residuals at a volatile solids (VS) concentration of approximately 40 g l(-1). At a hydraulic retention time (HRT) of 15 days and a VS loading rate of 2.7 g VS l(-1) day(-1), the mesophilic bioreactor showed an average specific volumetric biogas production rate of 0.88 l CH4 l(-1) day(-1) and an effluent VS concentration of 22.2 g VS l(-1) (43.0% VS removal efficiency) while the thermophilic bioreactor displayed similar performances. The overall methane yield for both systems was 0.21 l CH4 g(-1) VS fed and 0.47-0.48 l CH4 g(-1) VS removed. A primary limitation of thermophilic digestion of this protein-rich waste is the inhibition of methanogens due to higher nondissociated (free) ammonia (NH3) concentrations under similar total ammonium (NH4+) concentrations at equilibrium. Since thermophilic AD did not result in advantageous methane production rates or yields, mesophilic AD was, therefore, superior in treating secondary residuals from high-rate AD effluent. An additional digester to convert secondary residuals to methane may increase the total biogas generation at the brewery by 8% compared to just conventional high-rate digestion of brewery wastewater alone. PMID:18188623

  20. Pessimal shapes for packing

    NASA Astrophysics Data System (ADS)

    Kallus, Yoav

    2014-03-01

    The question of which convex shapes leave the most empty space in their densest packing is the subject of Reinhardt's conjecture in two dimensions and Ulam's conjecture in three dimensions. Such conjectures about pessimal packing shapes have proven notoriously difficult to make progress on. I show that the regular heptagon is a local pessimum among all convex shapes, and that the 3D ball is a local pessimum among origin-symmetric shapes. Any shape sufficiently close in the space of shapes to these local pessima can be packed at a greater efficiency than they. In two dimensions and in dimensions above three, the ball is not a local pessimum, so the situation in 3D is unusual and intriguing. I will discuss what conditions conspire to make the 3D ball a local pessimum and whether we can prove that it is also a global pessimum.

  1. The packing of particles

    SciTech Connect

    Cumberland, D.J.; Crawford, R.J.

    1987-01-01

    The wide range of information currently available on the packing of particles is brought together in this monograph. The authors' interest in the subject was initially aroused by the question of whether there is an optimum particle size distribution which would maximise the packing density of particles - a question which has attracted the interest of scientists and engineers for centuries. The densification of a powder mass is of relevance in a great many industries, among them the pharmaceutical, ceramic, powder metallurgy and civil engineering industries. In addition, the packing of regular - or irregular - shaped particles is also of relevance to a surprisingly large number of other industries and subject areas, i.e. the foundry industry, nuclear engineering, chemical engineering, crystallography, geology, biology, telecommunications, and so on. Accordingly, this book is written for a wide audience.

  2. Tissue grown in NASA Bioreactor

    NASA Technical Reports Server (NTRS)

    1998-01-01

    Cells from kidneys lose some of their special features in conventional culture but form spheres replete with specialized cell microvilli (hair) and synthesize hormones that may be clinically useful. Ground-based research studies have demonstrated that both normal and neoplastic cells and tissues recreate many of the characteristics in the NASA bioreactor that they display in vivo. Proximal kidney tubule cells that normally have rich apically oriented microvilli with intercellular clefts in the kidney do not form any of these structures in conventional two-dimensional monolayer culture. However, when normal proximal renal tubule cells are cultured in three-dimensions in the bioreactor, both the microvilli and the intercellular clefts form. This is important because, when the morphology is recreated, the function is more likely also to be rejuvenated. The work is sponsored by NASA's Office of Biological and Physical Research. The bioreactor is managed by the Biotechnology Cell Science Program at NASA's Johnson Space Center (JSC).

  3. Biological Hydrogen Production Using Chloroform-treated Methanogenic Granules

    NASA Astrophysics Data System (ADS)

    Hu, Bo; Chen, Shulin

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

  4. Reductive dechlorination of chlorophenols in methanogenic cultures

    SciTech Connect

    Wang, Y.T.; Muthukrishnan, S.; Wang, Z.

    1998-03-01

    Anaerobic biodegradation of a group of 12 chlorophenols (2-CP; 3-CP; 4-CP; 2,3-DCP; 2,6-DCP; 3,4-DCP; 3,5-DCP; 2,3,6-TCP; 2,4,5-DCP; 2,3,4,6-TeCP; 2,3,5,6-TeCP; and PCP) was examined in an unacclimated digester sludge culture and in a phenol-enriched, 2-CP and 3-CP acclimated methanogenic culture. The phenol-enriched culture degraded all chlorophenols except 4-CP, whereas the digester sludge culture did not degrade PCP or 2,3,4,6-TeCP. Dechlorination products were observed in the phenol-enriched culture but not in the fresh sludge. Based on the observed dechlorination products, degradation pathways for chlorophenols in the phenol-enriched culture were proposed. The phenol-enriched culture degraded chlorophenols at much higher rates than the digester sludge by dechlorinating at the ortho and meta positions. In both cultures, the rate of degradation depended on the number as well as the ring position of chlorine substituents. Higher rates were generally observed with ortho dechlorination and for compounds with lower numbers of chlorine substituents. The average rate of degradation did not vary significantly between mono- and dichlorophenols or between tri- and tetrachlorophenols. Analysis with the Haldane expression revealed that the phenol-enriched culture was less susceptible to inhibition caused by chlorophenols and that it possessed a higher affinity for substrate than the digester sludge culture.

  5. Zinc deprivation of methanol fed anaerobic granular sludge bioreactors

    PubMed Central

    Fermoso, Fernando G.; Collins, Gavin; Bartacek, Jan

    2008-01-01

    The effect of omitting zinc from the influent of mesophilic (30 °C) methanol fed upflow anaerobic sludge bed (UASB) reactors, and latter zinc supplementation to the influent to counteract the deprivation, was investigated by coupling the UASB reactor performance to the microbial ecology of the bioreactor sludge. Limitation of the specific methanogenic activity (SMA) on methanol due to the absence of zinc from the influent developed after 137 days of operation. At that day, the SMA in medium with a complete trace metal solution except Zn was 3.4 g CH4-COD g VSS−1 day−1, compared to 4.2 g CH4-COD g VSS−1 day−1 in a medium with a complete (including zinc) trace metal solution. The methanol removal capacity during these 137 days was 99% and no volatile fatty acids accumulated. Two UASB reactors, inoculated with the zinc-deprived sludge, were operated to study restoration of the zinc limitation by zinc supplementation to the bioreactor influent. In a first reactor, no changes to the operational conditions were made. This resulted in methanol accumulation in the reactor effluent after 12 days of operation, which subsequently induced acetogenic activity 5 days after the methanol accumulation started. Methanogenesis could not be recovered by the continuous addition of 0.5 μM ZnCl2 to the reactor for 13 days. In the second reactor, 0.5 μM ZnCl2 was added from its start-up. Although the reactor stayed 10 days longer methanogenically than the reactor operated without zinc, methanol accumulation was observed in this reactor (up to 1.1 g COD-MeOH L−1) as well. This study shows that zinc limitation can induce failure of methanol fed UASB reactors due to acidification, which cannot be restored by resuming the continuous supply of the deprived metal. PMID:18283507

  6. Micro-scale H2-CO2 Dynamics in a Hydrogenotrophic Methanogenic Membrane Reactor.

    PubMed

    Garcia-Robledo, Emilio; Ottosen, Lars D M; Voigt, Niels V; Kofoed, M W; Revsbech, Niels P

    2016-01-01

    Biogas production is a key factor in a sustainable energy supply. It is possible to get biogas with very high methane content if the biogas reactors are supplied with exogenous hydrogen, and one of the technologies for supplying hydrogen is through gas permeable membranes. In this study the activity and stratification of hydrogen consumption above such a membrane was investigated by use of microsensors for hydrogen and pH. A hydrogenotrophic methanogenic community that was able to consume the hydrogen flux within 0.5 mm of the membrane with specific rates of up to 30 m(3) H2 m(-3) day(-1) developed within 3 days in fresh manure and was already established at time zero when analyzing slurry from a biogas plant. The hydrogen consumption was dependent on a simultaneous carbon dioxide supply and was inhibited when carbon dioxide depletion elevated the pH to 9.2. The activity was only partially restored when the carbon dioxide supply was resumed. Bioreactors supplied with hydrogen gas should thus be carefully monitored and either have the hydrogen supply disrupted or be supplemented with carbon dioxide when the pH rises to values about 9. PMID:27582736

  7. Micro-scale H2–CO2 Dynamics in a Hydrogenotrophic Methanogenic Membrane Reactor

    PubMed Central

    Garcia-Robledo, Emilio; Ottosen, Lars D. M.; Voigt, Niels V.; Kofoed, M. W.; Revsbech, Niels P.

    2016-01-01

    Biogas production is a key factor in a sustainable energy supply. It is possible to get biogas with very high methane content if the biogas reactors are supplied with exogenous hydrogen, and one of the technologies for supplying hydrogen is through gas permeable membranes. In this study the activity and stratification of hydrogen consumption above such a membrane was investigated by use of microsensors for hydrogen and pH. A hydrogenotrophic methanogenic community that was able to consume the hydrogen flux within 0.5 mm of the membrane with specific rates of up to 30 m3 H2 m-3 day-1 developed within 3 days in fresh manure and was already established at time zero when analyzing slurry from a biogas plant. The hydrogen consumption was dependent on a simultaneous carbon dioxide supply and was inhibited when carbon dioxide depletion elevated the pH to 9.2. The activity was only partially restored when the carbon dioxide supply was resumed. Bioreactors supplied with hydrogen gas should thus be carefully monitored and either have the hydrogen supply disrupted or be supplemented with carbon dioxide when the pH rises to values about 9. PMID:27582736

  8. Use Alkalinity Monitoring to Optimize Bioreactor Performance.

    PubMed

    Jones, Christopher S; Kult, Keegan J

    2016-05-01

    In recent years, the agricultural community has reduced flow of nitrogen from farmed landscapes to stream networks through the use of woodchip denitrification bioreactors. Although deployment of this practice is becoming more common to treat high-nitrate water from agricultural drainage pipes, information about bioreactor management strategies is sparse. This study focuses on the use of water monitoring, and especially the use of alkalinity monitoring, in five Iowa woodchip bioreactors to provide insights into and to help manage bioreactor chemistry in ways that will produce desirable outcomes. Results reported here for the five bioreactors show average annual nitrate load reductions between 50 and 80%, which is acceptable according to established practice standards. Alkalinity data, however, imply that nitrous oxide formation may have regularly occurred in at least three of the bioreactors that are considered to be closed systems. Nitrous oxide measurements of influent and effluent water provide evidence that alkalinity may be an important indicator of bioreactor performance. Bioreactor chemistry can be managed by manipulation of water throughput in ways that produce adequate nitrate removal while preventing undesirable side effects. We conclude that (i) water should be retained for longer periods of time in bioreactors where nitrous oxide formation is indicated, (ii) measuring only nitrate and sulfate concentrations is insufficient for proper bioreactor operation, and (iii) alkalinity monitoring should be implemented into protocols for bioreactor management. PMID:27136151

  9. Use Alkalinity Monitoring to Optimize Bioreactor Performance.

    PubMed

    Jones, Christopher S; Kult, Keegan J

    2016-05-01

    In recent years, the agricultural community has reduced flow of nitrogen from farmed landscapes to stream networks through the use of woodchip denitrification bioreactors. Although deployment of this practice is becoming more common to treat high-nitrate water from agricultural drainage pipes, information about bioreactor management strategies is sparse. This study focuses on the use of water monitoring, and especially the use of alkalinity monitoring, in five Iowa woodchip bioreactors to provide insights into and to help manage bioreactor chemistry in ways that will produce desirable outcomes. Results reported here for the five bioreactors show average annual nitrate load reductions between 50 and 80%, which is acceptable according to established practice standards. Alkalinity data, however, imply that nitrous oxide formation may have regularly occurred in at least three of the bioreactors that are considered to be closed systems. Nitrous oxide measurements of influent and effluent water provide evidence that alkalinity may be an important indicator of bioreactor performance. Bioreactor chemistry can be managed by manipulation of water throughput in ways that produce adequate nitrate removal while preventing undesirable side effects. We conclude that (i) water should be retained for longer periods of time in bioreactors where nitrous oxide formation is indicated, (ii) measuring only nitrate and sulfate concentrations is insufficient for proper bioreactor operation, and (iii) alkalinity monitoring should be implemented into protocols for bioreactor management.

  10. Sun Packs Double Punch

    NASA Video Gallery

    On August 3, the sun packed a double punch, emitting a M6.0-class flare at 9:43 am EDT. This video is of the second, slightly stronger M9.3-class flare at 11:41 pm EDT. Both flares had significant ...

  11. Packing Them In.

    ERIC Educational Resources Information Center

    Carter, Claudia

    1997-01-01

    This activity involves students investigating the mathematics of packaging and exploring various concepts in geometry, including area and the Pythagorean theorem. Mathematics comes out of the discussion of packaging cans into six-packs and focuses on the cost-effectiveness of the horizontal storage area used. Students learn how knowledge of…

  12. Nutrition Action Pack.

    ERIC Educational Resources Information Center

    Sockut, Joanne; Stumpe, Stephanie

    One of five McDonald's Action Packs, these instructional materials integrate elementary school-level nutrition education into other disciplines--biology, sociology, physiology, mathematics, and art. Contents include four units consisting of twelve activities. Unit 1, Why You Need Food, is a self-examination of what is needed for growth, health,…

  13. Economics Action Pack.

    ERIC Educational Resources Information Center

    McDonald's Corp., Oak Brook, IL.

    One of five McDonald's Action Packs, this learning package introduces intermediate grade students to basic economic concepts. The fourteen activities include the topics of consumption (4 activities), production (5), the market system (3), a pretest, and a posttest. Specific titles under consumption include The Wonderful Treasure Tree (introduction…

  14. AUTOmatic Message PACKing Facility

    2004-07-01

    AUTOPACK is a library that provides several useful features for programs using the Message Passing Interface (MPI). Features included are: 1. automatic message packing facility 2. management of send and receive requests. 3. management of message buffer memory. 4. determination of the number of anticipated messages from a set of arbitrary sends, and 5. deterministic message delivery for testing purposes.

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

    NASA Astrophysics Data System (ADS)

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

    2015-12-01

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

  16. Two new disposable bioreactors for plant cell culture: The wave and undertow bioreactor and the slug bubble bioreactor.

    PubMed

    Terrier, Bénédicte; Courtois, Didier; Hénault, Nicolas; Cuvier, Arnaud; Bastin, Maryse; Aknin, Aziz; Dubreuil, Julien; Pétiard, Vincent

    2007-04-01

    The present article describes two novel flexible plastic-based disposable bioreactors. The first one, the WU bioreactor, is based on the principle of a wave and undertow mechanism that provides agitation while offering convenient mixing and aeration to the plant cell culture contained within the bioreactor. The second one is a high aspect ratio bubble column bioreactor, where agitation and aeration are achieved through the intermittent generation of large diameter bubbles, "Taylor-like" or "slug bubbles" (SB bioreactor). It allows an easy volume increase from a few liters to larger volumes up to several hundred liters with the use of multiple units. The cultivation of tobacco and soya cells producing isoflavones is described up to 70 and 100 L working volume for the SB bioreactor and WU bioreactor, respectively. The bioreactors being disposable and pre-sterilized before use, cleaning, sterilization, and maintenance operations are strongly reduced or eliminated. Both bioreactors represent efficient and low cost cell culture systems, applicable to various cell cultures at small and medium scale, complementary to traditional stainless-steel bioreactors.

  17. Biological hydrogen production using a membrane bioreactor.

    PubMed

    Oh, Sang-Eun; Iyer, Prabha; Bruns, Mary Ann; Logan, Bruce E

    2004-07-01

    A cross-flow membrane was coupled to a chemostat to create an anaerobic membrane bioreactor (MBR) for biological hydrogen production. The reactor was fed glucose (10,000 mg/L) and inoculated with a soil inoculum heat-treated to kill non-spore-forming methanogens. Hydrogen gas was consistently produced at a concentration of 57-60% in the headspace under all conditions. When operated in chemostat mode (no flow through the membrane) at a hydraulic retention time (HRT) of 3.3 h, 90% of the glucose was removed, producing 2200 mg/L of cells and 500 mL/h of biogas. When operated in MBR mode, the solids retention time (SRT) was increased to SRT = 12 h producing a solids concentration in the reactor of 5800 mg/L. This SRT increased the overall glucose utilization (98%), the biogas production rate (640 mL/h), and the conversion efficiency of glucose-to-hydrogen from 22% (no MBR) to 25% (based on a maximum of 4 mol-H(2)/mol-glucose). When the SRT was increased from 5 h to 48 h, glucose utilization (99%) and biomass concentrations (8,800 +/- 600 mg/L) both increased. However, the biogas production decreased (310 +/- 40 mL/h) and the glucose-to-hydrogen conversion efficiency decreased from 37 +/- 4% to 18 +/- 3%. Sustained permeate flows through the membrane were in the range of 57 to 60 L/m(2) h for three different membrane pore sizes (0.3, 0.5, and 0.8 microm). Most (93.7% to 99.3%) of the membrane resistance was due to internal fouling and the reversible cake resistance, and not the membrane itself. Regular backpulsing was essential for maintaining permeate flux through the membrane. Analysis of DNA sequences using ribosomal intergenic spacer analysis indicated bacteria were most closely related to members of Clostridiaceae and Flexibacteraceae, including Clostridium acidisoli CAC237756 (97%), Linmingia china AF481148 (97%), and Cytophaga sp. MDA2507 AF238333 (99%). No PCR amplification of 16s rRNA genes was obtained when archaea-specific primers were used.

  18. Hydrogen transfer between methanogens and fermentative heterotrophs in hyperthermophilic cocultures

    SciTech Connect

    Muralidharan, V.; Hirsh, I.S.; Bouwer, E.J.; Rinker, K.D.; Kelly, R.M.

    1997-11-05

    Interactions involving hydrogen transfer were studied in a coculture of two hyperthermophilic microorganisms: Thermotoga maritima, an anaerobic heterotroph, and Methanococcus jannaschii, a hydrogenotrophic methanogen. Cell densities of T. maritima increased 10-fold when cocultured with M. jannaschii at 85 C, and the methanogen was able to grow in the absence of externally supplied H{sub 2} and CO{sub 2}. The coculture could not be established if the two organisms were physically separated by a dialysis membrane, suggesting the importance of spatial proximity. The significance of spatial proximity was also supported by cell cytometry, where the methanogen was only found in cell sorts at or above 4.5 {micro}m in samples of the coculture in exponential phase. An unstructured mathematical model was used to compare the influence of hydrogen transport and metabolic properties on mesophilic and hyperthermophilic cocultures. Calculations suggest the increases in methanogenesis rates with temperature result from greater interactions between the methanogenic and fermentative organisms, as evidenced by the sharp decline in H{sub 2} concentration in the proximity of a hyperthermophilic methanogen. The experimental and modeling results presented here illustrate the need to consider the interactions within hyperthermophilic consortia when choosing isolation strategies and evaluating biotransformations at elevated temperatures.

  19. [Methanogens and manipulation of methane production in the rumen].

    PubMed

    Guo, Yan-qiu; Hu, Wei-lian; Liu, Jian-xin

    2005-02-01

    Methanogens belong to the kingdom of Euryarchaeota in the domain of Archaea. They are characterized by their ability to produce methane under anaerobic conditions. Methane production in the rumen represents a loss of energy for the host animal, and, in addition, methane eructated by ruminants may contribute to a greenhouse effect or global warming. Reduction or elimination of methanogenesis in the rumen has been touted as a way of improving animal production and may marginally benefit to control of anthropogenic release of methane. More and more scientists focus on ruminal methanogens and methanogenesis recently. Authors summarized the manipulation of methanogenesis in the rumen, including defaunation, feed formulation, adding electron acceptors and stimulation of acetogens. The characteristics of methanogenic Archaea and the recent knowledge of the methanogenesis in the rumen were also reviewed in this article.

  20. Growth of methanogens on a Mars soil simulant.

    PubMed

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

    2004-12-01

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

  1. PRACTICE REVIEW OF FIVE BIOREACTOR/RECIRCULATION LANDFILLS

    EPA Science Inventory

    Six bioreactor landfills were analyzed to provide a perspective of current practice and technical issues that differentiate bioreactor landfills from conventional landfills. Five of the bioreactor landfills were anaerobic and one was aerated. In one case, nearly identical cells e...

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

    SciTech Connect

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

    1999-02-15

    Radiometric detection headspace analyses of microcosms containing bed sediments from two geographically distinct sites indicated that 10--39% of the radiolabeled carbon transformed during anaerobic biodegradation of [1,2-{sup 14}C]trichloroethene (TCE) or [1,2-{sup 14}C]vinyl chloride (VC) under methanogenic conditions was ultimately incorporated into {sup 14}CH{sub 4}. The results demonstrate that, in addition to ethene, ethane, and CO{sub 2}, CH{sub 4} can be a significant product of chloroethene biodegradation in some methanogenic sediments.

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

    NASA Technical Reports Server (NTRS)

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

    1977-01-01

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

  4. Syntrophic Degradation of Lactate in Methanogenic Co-cultures

    SciTech Connect

    Meyer, Birte; Stahl, David

    2010-05-17

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

  5. Inhibitory Effects of Ferrihydrite on a Thermophilic Methanogenic Community

    PubMed Central

    Yamada, Chihaya; Kato, Souichiro; Ueno, Yoshiyuki; Ishii, Masaharu; Igarashi, Yasuo

    2014-01-01

    The addition of ferrihydrite to methanogenic microbial communities obtained from a thermophilic anaerobic digester suppressed methanogenesis in a dose-dependent manner. The amount of reducing equivalents consumed by the reduction of iron was significantly smaller than that expected from the decrease in the production of CH4, which suggested that competition between iron-reducing microorganisms and methanogens was not the most significant cause for the suppression of methanogenesis. Microbial community analyses revealed that the presence of ferrihydrite markedly affected the bacterial composition, but not the archaeal composition. These results indicate that the presence of ferrihydrite directly and indirectly suppresses thermophilic methanogenesis. PMID:24859310

  6. Conversion of Cn-Unsaturated into Cn-2-Saturated LCFA Can Occur Uncoupled from Methanogenesis in Anaerobic Bioreactors.

    PubMed

    Cavaleiro, Ana J; Pereira, Maria Alcina; Guedes, Ana P; Stams, Alfons J M; Alves, M Madalena; Sousa, Diana Z

    2016-03-15

    Fat, oils, and grease present in complex wastewater can be readily converted to methane, but the energy potential of these compounds is not always recyclable, due to incomplete degradation of long chain fatty acids (LCFA) released during lipids hydrolysis. Oleate (C18:1) is generally the dominant LCFA in lipid-containing wastewater, and its conversion in anaerobic bioreactors results in palmitate (C16:0) accumulation. The reason why oleate is continuously converted to palmitate without further degradation via β-oxidation is still unknown. In this work, the influence of methanogenic activity in the initial conversion steps of unsaturated LCFA was studied in 10 bioreactors continuously operated with saturated or unsaturated C16- and C18-LCFA, in the presence or absence of the methanogenic inhibitor bromoethanesulfonate (BrES). Saturated Cn-2-LCFA accumulated both in the presence and absence of BrES during the degradation of unsaturated Cn-LCFA, and represented more than 50% of total LCFA. In the presence of BrES further conversion of saturated intermediates did not proceed, not even when prolonged batch incubation was applied. As the initial steps of unsaturated LCFA degradation proceed uncoupled from methanogenesis, accumulation of saturated LCFA can be expected. Analysis of the active microbial communities suggests a role for facultative anaerobic bacteria in the initial steps of unsaturated LCFA biodegradation. Understanding this role is now imperative to optimize methane production from LCFA. PMID:26810160

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

    PubMed Central

    Fu, Li; Song, Tianze; Lu, Yahai

    2015-01-01

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

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

    PubMed

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

    2016-01-01

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

  9. Methane production in simulated hybrid bioreactor landfill.

    PubMed

    Xu, Qiyong; Jin, Xiao; Ma, Zeyu; Tao, Huchun; Ko, Jae Hac

    2014-09-01

    The aim of this work was to study a hybrid bioreactor landfill technology for landfill methane production from municipal solid waste. Two laboratory-scale columns were operated for about ten months to simulate an anaerobic and a hybrid landfill bioreactor, respectively. Leachate was recirculated into each column but aeration was conducted in the hybrid bioreactor during the first stage. Results showed that leachate pH in the anaerobic bioreactor maintained below 6.5, while in the hybrid bioreactor quickly increased from 5.6 to 7.0 due to the aeration. The temporary aeration resulted in lowering COD and BOD5 in the leachate. The volume of methane collected from the hybrid bioreactor was 400 times greater than that of the anaerobic bioreactor. Also, the methane production rate of the hybrid bioreactor was improved within a short period of time. After about 10 months' operation, the total methane production in the hybrid bioreactor was 212 L (16 L/kgwaste).

  10. Prostate tumor grown in NASA Bioreactor

    NASA Technical Reports Server (NTRS)

    2001-01-01

    This prostate cancer construct was grown during NASA-sponsored bioreactor studies on Earth. Cells are attached to a biodegradable plastic lattice that gives them a head start in growth. Prostate tumor cells are to be grown in a NASA-sponsored Bioreactor experiment aboard the STS-107 Research-1 mission in 2002. Dr. Leland Chung of the University of Virginia is the principal investigator. The NASA Bioreactor provides a low turbulence culture environment which promotes the formation of large, three-dimensional cell clusters. Due to their high level of cellular organization and specialization, samples constructed in the bioreactor more closely resemble the original tumor or tissue found in the body. The Bioreactor is rotated to provide gentle mixing of fresh and spent nutrient without inducing shear forces that would damage the cells. The work is sponsored by NASA's Office of Biological and Physical Research. The bioreactor is managed by the Biotechnology Cell Science Program at NASA's Johnson Space Center (JSC). NASA-sponsored bioreactor research has been instrumental in helping scientists to better understand normal and cancerous tissue development. In cooperation with the medical community, the bioreactor design is being used to prepare better models of human colon, prostate, breast and ovarian tumors. Cartilage, bone marrow, heart muscle, skeletal muscle, pancreatic islet cells, liver and kidney are just a few of the normal tissues being cultured in rotating bioreactors by investigators. Credit: NASA and the University of Virginia.

  11. Monolithic Continuous-Flow Bioreactors

    NASA Technical Reports Server (NTRS)

    Stephanopoulos, Gregory; Kornfield, Julia A.; Voecks, Gerald A.

    1993-01-01

    Monolithic ceramic matrices containing many small flow passages useful as continuous-flow bioreactors. Ceramic matrix containing passages made by extruding and firing suitable ceramic. Pores in matrix provide attachment medium for film of cells and allow free movement of solution. Material one not toxic to micro-organisms grown in reactor. In reactor, liquid nutrients flow over, and liquid reaction products flow from, cell culture immobilized in one set of channels while oxygen flows to, and gaseous reaction products flow from, culture in adjacent set of passages. Cells live on inner surfaces containing flowing nutrient and in pores of walls of passages. Ready access to nutrients and oxygen in channels. They generate continuous high yield characteristic of immobilized cells, without large expenditure of energy otherwise incurred if necessary to pump nutrient solution through dense biomass as in bioreactors of other types.

  12. Review of nonconventional bioreactor technology

    SciTech Connect

    Turick, C.E.; Mcllwain, M.E.

    1993-09-01

    Biotechnology will significantly affect many industrial sectors in the future. Industrial sectors that will be affected include pharmaceutical, chemical, fuel, agricultural, and environmental remediation. Future research is needed to improve bioprocessing efficiency and cost-effectiveness in order to compete with traditional technologies. This report describes recent advances in bioprocess technologies and bioreactor designs and relates them to problems encountered in many industrial bioprocessing operations. The primary focus is directed towards increasing gas and vapor transfer for enhanced bioprocess kinetics as well as unproved by-product separation and removal. The advantages and disadvantages of various conceptual designs such as hollow-fiber, gas-phase, hyperbaric/hypobaric, and electrochemical bioreactors are also discussed. Specific applications that are intended for improved bioprocesses include coal desulfurization, coal liquefaction, soil bioremediation, biomass conversion to marketable chemicals, biomining, and biohydrometallurgy as well as bioprocessing of gases and vapors.

  13. A high-rate perfusion bioreactor for plant cells.

    PubMed

    De Dobbeleer, C; Cloutier, M; Fouilland, M; Legros, R; Jolicoeur, M

    2006-12-20

    A perfusion bioreactor allowing continuous extraction of secondary metabolites was designed and challenged for Eschscholtzia californica plant cell suspensions. Four sedimentation columns mounted inside a 2.5-L bioreactor separated single cells and cell aggregates from the culture medium. Cells were elicited with chitin at day 4 and the liquid medium free of cells and debris was then continuously pumped to the extraction columns containing fluidized XAD-7 resins, and then recirculated back to the cell suspension. A medium upward velocity corresponding to cell sedimentation velocity maintained a stable cell/medium separation front in the columns for sedimented cell volume (SCV) of 90% (70% packed cell volume, PCV). Two perfusion bioreactor cultures of 10 and 14 days were performed. A maximum dilution rate of 20.4/day was reached from day 4 to day 6, and was then reduced to 5/day at day 9 for 55% SCV. Control cultures were performed without and with free extraction resins into the cell suspension. Perfusion cultures showed similar specific growth rates of 0.24 +/- 0.04/day before and after elicitation. However, production level in the perfusion cultures was similar to that from the culture without resins with a maximum of 2.06 micromole/gDW total alkaloids, with 1.54 micromole/gDW in the resins. Cultures with free resins resulted in 30.94 micromole/gDW with 28.4 +/- 8.8 micromole/gDW in the resins. Difference in the cells nutritional state from elicitation was identified as a major cause in the production reduction. However, pathway to chelilutine was favored in the continuous extraction culture.

  14. Oxygen supply for CHO cells immobilized on a packed-bed of Fibra-Cel disks.

    PubMed

    Meuwly, F; Loviat, F; Ruffieux, P-A; Bernard, A R; Kadouri, A; von Stockar, U

    2006-03-01

    Packed-bed bioreactors (PBR) have proven to be efficient systems to culture mammalian cells at very high cell density in perfusion mode, thus leading to very high volumetric productivity. However, the immobilized cells must be continuously supplied with all nutrients in sufficient quantities to remain viable and productive over the full duration of the perfusion culture. Among all nutrients, oxygen is the most critical since it is present at very low concentration due to its low solubility in cell culture medium. This work presents the development of a model for oxygenation in a packed-bed bioreactor system. The experimental system used to develop the model was a packed-bed of Fibra-Cel disk carriers used to cultivate Chinese Hamster Ovary cells at high density ( approximately 6.1 x 10(7) cell/mL) in perfusion mode. With the help of this model, it was possible to identify if a PBR system is operated in optimal or sub-optimal conditions. Using the model, two options were proposed, which could improve the performance of the basal system by about twofold, that is, by increasing the density of immobilized cells per carrier volume from 6.1 x 10(7) to 1.2 x 10(8) cell/mL, or by increasing the packed-bed height from 0.2 to 0.4 m. Both strategies would be rather simple to test and implement in the packed-bed bioreactor system used for this study. As a result, it would be possible to achieve a substantial improvement of about twofold higher productivity as compared with the basal conditions.

  15. Bioreactor concepts for cell culture-based viral vaccine production.

    PubMed

    Gallo-Ramírez, Lilí Esmeralda; Nikolay, Alexander; Genzel, Yvonne; Reichl, Udo

    2015-01-01

    Vaccine manufacturing processes are designed to meet present and upcoming challenges associated with a growing vaccine market and to include multi-use facilities offering a broad portfolio and faster reaction times in case of pandemics and emerging diseases. The final products, from whole viruses to recombinant viral proteins, are very diverse, making standard process strategies hardly universally applicable. Numerous factors such as cell substrate, virus strain or expression system, medium, cultivation system, cultivation method, and scale need consideration. Reviewing options for efficient and economical production of human vaccines, this paper discusses basic factors relevant for viral antigen production in mammalian cells, avian cells and insect cells. In addition, bioreactor concepts, including static systems, single-use systems, stirred tanks and packed-beds are addressed. On this basis, methods towards process intensification, in particular operational strategies, the use of perfusion systems for high product yields, and steps to establish continuous processes are introduced.

  16. Relating methanogen community structure and anaerobic digester function.

    PubMed

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

    2015-03-01

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

  17. Dynamics of the Methanogenic Archaea in Tropical Estuarine Sediments

    PubMed Central

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

    2013-01-01

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

  18. Dynamics of the methanogenic archaea in tropical estuarine sediments.

    PubMed

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

    2013-01-01

    Methanogenesis may represent a key process in the terminal phases of anaerobic organic matter mineralization in sediments of coastal lagoons. The aim of the present work was to study the temporal and spatial dynamics of methanogenic archaea in sediments of tropical coastal lagoons and their relationship with environmental changes in order to determine how these influence methanogenic community. Sediment samples were collected during the dry (February, May, and early June) and rainy seasons (July, October, and November). Microbiological analysis included the quantification of viable methanogenic archaea (MA) with three substrates and the evaluation of kinetic activity from acetate in the presence and absence of sulfate. The environmental variables assessed were temperature, pH, Eh, salinity, sulfate, solids content, organic carbon, and carbohydrates. MA abundance was significantly higher in the rainy season (10(6)-10(7) cells/g) compared with the dry season (10(4)-10(6) cells/g), with methanol as an important substrate. At spatial level, MA were detected in the two layers analyzed, and no important variations were observed either in MA abundance or activity. Salinity, sulfate, solids, organic carbon, and Eh were the environmental variables related to methanogenic community. A conceptual model is proposed to explain the dynamics of the MA.

  19. Metabolic reconstruction of the archaeon methanogen Methanosarcina Acetivorans

    PubMed Central

    2011-01-01

    Background Methanogens are ancient organisms that are key players in the carbon cycle accounting for about one billion tones of biological methane produced annually. Methanosarcina acetivorans, with a genome size of ~5.7 mb, is the largest sequenced archaeon methanogen and unique amongst the methanogens in its biochemical characteristics. By following a systematic workflow we reconstruct a genome-scale metabolic model for M. acetivorans. This process relies on previously developed computational tools developed in our group to correct growth prediction inconsistencies with in vivo data sets and rectify topological inconsistencies in the model. Results The generated model iVS941 accounts for 941 genes, 705 reactions and 708 metabolites. The model achieves 93.3% prediction agreement with in vivo growth data across different substrates and multiple gene deletions. The model also correctly recapitulates metabolic pathway usage patterns of M. acetivorans such as the indispensability of flux through methanogenesis for growth on acetate and methanol and the unique biochemical characteristics under growth on carbon monoxide. Conclusions Based on the size of the genome-scale metabolic reconstruction and extent of validated predictions this model represents the most comprehensive up-to-date effort to catalogue methanogenic metabolism. The reconstructed model is available in spreadsheet and SBML formats to enable dissemination. PMID:21324125

  20. Sulfate reducers can outcompete methanogens at freshwater sulfate concentrations

    SciTech Connect

    Lovley, D.R.; Klug, M.J.

    1983-01-01

    Acetate and hydrogen metabolism by sulfate reducers and methanogens in the profundal sediments of an oligotrophic lake were examined. Inhibition of sulfate reduction with molybdate stimulated methane production from both hydrogen and acetate. Molybdate did not stimulate methane production in sediments that were preincubated to deplete the sulfate pool. Sulfate reduction accounted for 30 to 81% of the total of terminal metabolism proceeding through sulfate reduction and methane production in Eckman grab samples of surface sediments. The ability of sulfate reducers to effectively compete with methanogens for acetate was related to the sulfate reducers lower half-saturation constant for acetate metabolism at in situ sulfate concentrations. Processes other than sulfate reduction and methanogenesis consumed hydrogen at elevated hydrogen partial pressures and prevented a kinetic analysis of hydrogen uptake by sulfate reducers and methanogens. The demonstration that sulfate reducers can successfully compete with methanogens for hydrogen and acetate in sediments at in situ sulfate concentrations of 60 to 105 mM extends the known range of sediment habitats in which sulfate reduction can be a dominant terminal process.

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

    EPA Science Inventory

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

  2. Effect of polyvinyl alcohol hydrogel as a biocarrier on volatile fatty acids production of a two-stage thermophilic anaerobic membrane bioreactor.

    PubMed

    Chaikasem, Supawat; Abeynayaka, Amila; Visvanathan, Chettiyappan

    2014-09-01

    This work studied the effect of polyvinyl alcohol hydrogel (PVA-gel) beads, as an effective biocarrier for volatile fatty acid (VFA) production in hydrolytic reactor of a two-stage thermophilic anaerobic membrane bioreactor (TAnMBR). The two-stage TAnMBR, treating synthetic high strength particulate wastewater with influent chemical oxygen demand (COD) [16.4±0.8 g/L], was operated at 55 °C. Under steady state conditions, the reactor was operated at an organic loading rate of 8.2±0.4 kg COD/m(3) d. Operational performance of the system was monitored by assessing VFA composition and quantity, methane production and COD removal efficiency. Increment of VFA production was observed with PVA-gel addition. Hydrolytic effluent contained large amount of acetic acid and n-butyric acid. However, increase in VFA production adversely affected the methanogenic reactor performance due to lack of methanogenic archaea.

  3. Anaerobic degradation of phthalate isomers by methanogenic consortia

    SciTech Connect

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

    1999-03-01

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

  4. Estimation of methanogen biomass by quantitation of coenzyme M

    SciTech Connect

    Elias, D.A.; Krumholz, L.R.; Tanner, R.S.; Suflita, J.M.

    1999-12-01

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

  5. Performance linked to residence time distribution by a novel wool-based bioreactor for tertiary sewage treatment.

    PubMed

    Hu, Bibo; Wheatley, Andrew; Ishtchenko, Vera; Huddersman, Katherine

    2012-05-01

    Laboratory-scale experiments were carried out using up-flow 7 L Submerged Aerated Filter reactors packed with wool fibre or commercial plastic pall rings, Kaldnes, (70% by volume) support media for the tertiary treatment of sewage. The performance of the wool bioreactor was more consistent than that with Kaldnes medium, for both TOC removal (93%) and SS removal (90%). Both plastic and wool-packed bioreactors achieved complete nitrification at the load of about 0.4 kgCOD/m(3)/day. The sludge yield of the wool bioreactor was almost half that of the bioreactor with Kaldnes suggesting that wool could retain residual organics and particulates. The wool however was degraded and it was concluded that wool would have to be considered as additional sacrificial adsorption capacity rather than an alternative medium. The performance was linked to the residence time distribution studies and these changes in the wool structure. Biomass growth increased the retention of the tracer in the wool reactor by, it was suggested, exposing a greater surface area. Results from the plastic media on the other hand showed increased mixing possibly by increasing the mobility of the plastic. Aeration increased the mixing in both reactors, and patterns were in all cases predominantly well-mixed.

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

    DOE PAGES

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

    2014-09-04

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

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

    PubMed

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

    2016-07-13

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

  8. BIOREACTOR LANDFILLS, THEORETICAL ADVANTAGES AND RESEARCH CHALLENGES

    EPA Science Inventory

    Bioreactor landfills are municipal solid waste landfills that utilize bulk liquids in an effort to accelerate solid waste degradation. There are few potential benefits for operating a MSW landfill as a bioreactor. These include leachate treatment and management, increase in the s...

  9. BIOREACTOR DESIGN - OUTER LOOP LANDFILL, LOUISVILLE, KY

    EPA Science Inventory

    Bioreactor field demonstration projects are underway at the Outer Loop Landfill in Louisville, KY, USA. The research effort is a cooperative research effort between US EPA and Waste Management Inc. Two primary kinds of municipal waste bioreactors are under study at this site. ...

  10. Bioreactors for Plant Embryogenesis and Beyond.

    PubMed

    Fei, Liwen; Weathers, Pamela

    2016-01-01

    A variety of different bioreactors have been developed for use in initiating and cultivating somatic embryos. The various designs for embryogenesis and culture are critically evaluated here. Bioreactor optimization and operation methods are also described along with recommendations for use based on desired outcome.

  11. Membrane biofilm development improves COD removal in anaerobic membrane bioreactor wastewater treatment.

    PubMed

    Smith, Adam L; Skerlos, Steven J; Raskin, Lutgarde

    2015-09-01

    Membrane biofilm development was evaluated to improve psychrophilic (15°C) anaerobic membrane bioreactor (AnMBR) treatment of domestic wastewater. An AnMBR containing three replicate submerged membrane housings with separate permeate collection was operated at three levels of membrane fouling by independently controlling biogas sparging for each membrane unit. High membrane fouling significantly improved permeate quality, but resulted in dissolved methane in the permeate at a concentration two to three times the equilibrium concentration predicted by Henry's law. Illumina sequencing of 16S rRNA targeting Bacteria and Archaea and reverse transcription-quantitative polymerase chain reaction targeting the methyl coenzyme-M reductase (mcrA) gene in methanogens indicated that the membrane biofilm was enriched in highly active methanogens and syntrophic bacteria. Restoring fouled membranes to a transmembrane pressure (TMP) near zero by increasing biogas sparging did not disrupt the biofilm's treatment performance, suggesting that microbes in the foulant layer were tightly adhered and did not significantly contribute to TMP. Dissolved methane oversaturation persisted without high TMP, implying that methanogenesis in the biofilm, rather than high TMP, was the primary driving force in methane oversaturation. The results describe an attractive operational strategy to improve treatment performance in low-temperature AnMBR by supporting syntrophy and methanogenesis in the membrane biofilm through controlled membrane fouling. PMID:26238293

  12. Membrane biofilm development improves COD removal in anaerobic membrane bioreactor wastewater treatment.

    PubMed

    Smith, Adam L; Skerlos, Steven J; Raskin, Lutgarde

    2015-09-01

    Membrane biofilm development was evaluated to improve psychrophilic (15°C) anaerobic membrane bioreactor (AnMBR) treatment of domestic wastewater. An AnMBR containing three replicate submerged membrane housings with separate permeate collection was operated at three levels of membrane fouling by independently controlling biogas sparging for each membrane unit. High membrane fouling significantly improved permeate quality, but resulted in dissolved methane in the permeate at a concentration two to three times the equilibrium concentration predicted by Henry's law. Illumina sequencing of 16S rRNA targeting Bacteria and Archaea and reverse transcription-quantitative polymerase chain reaction targeting the methyl coenzyme-M reductase (mcrA) gene in methanogens indicated that the membrane biofilm was enriched in highly active methanogens and syntrophic bacteria. Restoring fouled membranes to a transmembrane pressure (TMP) near zero by increasing biogas sparging did not disrupt the biofilm's treatment performance, suggesting that microbes in the foulant layer were tightly adhered and did not significantly contribute to TMP. Dissolved methane oversaturation persisted without high TMP, implying that methanogenesis in the biofilm, rather than high TMP, was the primary driving force in methane oversaturation. The results describe an attractive operational strategy to improve treatment performance in low-temperature AnMBR by supporting syntrophy and methanogenesis in the membrane biofilm through controlled membrane fouling.

  13. Membrane biofilm development improves COD removal in anaerobic membrane bioreactor wastewater treatment

    PubMed Central

    Smith, Adam L; Skerlos, Steven J; Raskin, Lutgarde

    2015-01-01

    Membrane biofilm development was evaluated to improve psychrophilic (15°C) anaerobic membrane bioreactor (AnMBR) treatment of domestic wastewater. An AnMBR containing three replicate submerged membrane housings with separate permeate collection was operated at three levels of membrane fouling by independently controlling biogas sparging for each membrane unit. High membrane fouling significantly improved permeate quality, but resulted in dissolved methane in the permeate at a concentration two to three times the equilibrium concentration predicted by Henry’s law. Illumina sequencing of 16S rRNA targeting Bacteria and Archaea and reverse transcription-quantitative polymerase chain reaction targeting the methyl coenzyme-M reductase (mcrA) gene in methanogens indicated that the membrane biofilm was enriched in highly active methanogens and syntrophic bacteria. Restoring fouled membranes to a transmembrane pressure (TMP) near zero by increasing biogas sparging did not disrupt the biofilm’s treatment performance, suggesting that microbes in the foulant layer were tightly adhered and did not significantly contribute to TMP. Dissolved methane oversaturation persisted without high TMP, implying that methanogenesis in the biofilm, rather than high TMP, was the primary driving force in methane oversaturation. The results describe an attractive operational strategy to improve treatment performance in low-temperature AnMBR by supporting syntrophy and methanogenesis in the membrane biofilm through controlled membrane fouling. PMID:26238293

  14. A vaccine against rumen methanogens can alter the composition of archaeal populations.

    PubMed

    Williams, Yvette J; Popovski, Sam; Rea, Suzanne M; Skillman, Lucy C; Toovey, Andrew F; Northwood, Korinne S; Wright, André-Denis G

    2009-04-01

    The objectives of this study were to formulate a vaccine based upon the different species/strains of methanogens present in sheep intended to be immunized and to determine if a targeted vaccine could be used to decrease the methane output of the sheep. Two 16S rRNA gene libraries were used to survey the methanogenic archaea in sheep prior to vaccination, and methanogens representing five phylotypes were found to account for >52% of the different species/strains of methanogens detected. A vaccine based on a mixture of these five methanogens was then formulated, and 32 sheep were vaccinated on days 0, 28, and 103 with either a control or the anti-methanogen vaccine. Enzyme-linked immunosorbent assay analysis revealed that each vaccination with the anti-methanogen formulation resulted in higher specific immunoglobulin G titers in plasma, saliva, and rumen fluid. Methane output levels corrected for dry-matter intake for the control and treatment groups were not significantly different, and real-time PCR data also indicated that methanogen numbers were not significantly different for the two groups after the second vaccination. However, clone library data indicated that methanogen diversity was significantly greater in sheep receiving the anti-methanogen vaccine and that the vaccine may have altered the composition of the methanogen population. A correlation between 16S rRNA gene sequence relatedness and cross-reactivity for the methanogens (R(2) = 0.90) also exists, which suggests that a highly specific vaccine can be made to target specific strains of methanogens and that a more broad-spectrum approach is needed for success in the rumen. Our data also suggest that methanogens take longer than 4 weeks to adapt to dietary changes and call into question the validity of experimental results based upon a 2- to 4-week acclimatization period normally observed for bacteria. PMID:19201957

  15. A vaccine against rumen methanogens can alter the composition of archaeal populations.

    PubMed

    Williams, Yvette J; Popovski, Sam; Rea, Suzanne M; Skillman, Lucy C; Toovey, Andrew F; Northwood, Korinne S; Wright, André-Denis G

    2009-04-01

    The objectives of this study were to formulate a vaccine based upon the different species/strains of methanogens present in sheep intended to be immunized and to determine if a targeted vaccine could be used to decrease the methane output of the sheep. Two 16S rRNA gene libraries were used to survey the methanogenic archaea in sheep prior to vaccination, and methanogens representing five phylotypes were found to account for >52% of the different species/strains of methanogens detected. A vaccine based on a mixture of these five methanogens was then formulated, and 32 sheep were vaccinated on days 0, 28, and 103 with either a control or the anti-methanogen vaccine. Enzyme-linked immunosorbent assay analysis revealed that each vaccination with the anti-methanogen formulation resulted in higher specific immunoglobulin G titers in plasma, saliva, and rumen fluid. Methane output levels corrected for dry-matter intake for the control and treatment groups were not significantly different, and real-time PCR data also indicated that methanogen numbers were not significantly different for the two groups after the second vaccination. However, clone library data indicated that methanogen diversity was significantly greater in sheep receiving the anti-methanogen vaccine and that the vaccine may have altered the composition of the methanogen population. A correlation between 16S rRNA gene sequence relatedness and cross-reactivity for the methanogens (R(2) = 0.90) also exists, which suggests that a highly specific vaccine can be made to target specific strains of methanogens and that a more broad-spectrum approach is needed for success in the rumen. Our data also suggest that methanogens take longer than 4 weeks to adapt to dietary changes and call into question the validity of experimental results based upon a 2- to 4-week acclimatization period normally observed for bacteria.

  16. Measuring Water in Bioreactor Landfills

    NASA Astrophysics Data System (ADS)

    Han, B.; Gallagher, V. N.; Imhoff, P. T.; Yazdani, R.; Chiu, P.

    2004-12-01

    Methane is an important greenhouse gas, and landfills are the largest anthropogenic source in many developed countries. Bioreactor landfills have been proposed as one means of abating greenhouse gas emissions from landfills. Here, the decomposition of organic wastes is enhanced by the controlled addition of water or leachate to maintain optimal conditions for waste decomposition. Greenhouse gas abatement is accomplished by sequestration of photosynthetically derived carbon in wastes, CO2 offsets from energy use of waste derived gas, and mitigation of methane emission from the wastes. Maintaining optimal moisture conditions for waste degradation is perhaps the most important operational parameter in bioreactor landfills. To determine how much water is needed and where to add it, methods are required to measure water within solid waste. However, there is no reliable method that can measure moisture content simply and accurately in the heterogeneous environment typical of landfills. While well drilling and analysis of solid waste samples is sometimes used to determine moisture content, this is an expensive, time-consuming, and destructive procedure. To overcome these problems, a new technology recently developed by hydrologists for measuring water in the vadose zone --- the partitioning tracer test (PTT) --- was evaluated for measuring water in solid waste in a full-scale bioreactor landfill in Yolo County, CA. Two field tests were conducted in different regions of an aerobic bioreactor landfill, with each test measuring water in ≈ 250 ft3 of solid waste. Tracers were injected through existing tubes inserted in the landfill, and tracer breakthrough curves were measured through time from the landfill's gas collection system. Gas samples were analyzed on site using a field-portable gas chromatograph and shipped offsite for more accurate laboratory analysis. In the center of the landfill, PTT measurements indicated that the fraction of the pore space filled with water

  17. Packing Products: Polystyrene vs. Cornstarch

    ERIC Educational Resources Information Center

    Starr, Suzanne

    2009-01-01

    Packing materials such as polystyrene take thousands of years to decompose, whereas packing peanuts made from cornstarch, which some companies are now using, can serve the same purpose, but dissolve in water. The author illustrates this point to her class one rainy day using the sculptures students made from polystyrene and with the cornstarch…

  18. Determination of methanogenic pathways through carbon isotope (δ13C) analysis for the two-stage anaerobic digestion of high-solids substrates.

    PubMed

    Gehring, Tito; Klang, Johanna; Niedermayr, Andrea; Berzio, Stephan; Immenhauser, Adrian; Klocke, Michael; Wichern, Marc; Lübken, Manfred

    2015-04-01

    This study used carbon isotope (δ(13)C)-based calculations to quantify the specific methanogenic pathways in a two-stage experimental biogas plant composed of three thermophilic leach bed reactors (51-56 °C) followed by a mesophilic (36.5 °C) anaerobic filter. Despite the continuous dominance of the acetoclastic Methanosaeta in the anaerobic filter, the methane (CH4) fraction derived from carbon dioxide reduction (CO2), fmc, varied significantly over the investigation period of 200 days. At organic loading rates (OLRs) below 6.0 gCOD L(-1) d(-1), the average fmc value was 33%, whereas at higher OLRs, with a maximum level of 17.0 gCOD L(-1) d(-1), the fmc values reached 47%. The experiments allowed for a clear differentiation of the isotope fractionation related to the formation and consumption of acetate in both stages of the plant. Our data indicate constant carbon isotope fractionation for acetate formation at different OLRs within the thermophilic leach bed reactors as well as a negligible contribution of homoacetogenesis. These results present the first quantification of methanogenic pathway (fmc values) dynamics for a continually operated mesophilic bioreactor and highlight the enormous potential of δ(13)C analysis for a more comprehensive understanding of the anaerobic degradation processes in CH4-producing biogas plants. PMID:25741999

  19. Phenotypic Properties and Microbial Diversity of Methanogenic Granules from a Full-Scale Upflow Anaerobic Sludge Bed Reactor Treating Brewery Wastewater†

    PubMed Central

    Díaz, Emiliano E.; Stams, Alfons J. M.; Amils, Ricardo; Sanz, José 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 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. PMID:16820491

  20. Diversity of bacteria, archaea and protozoa in a perchlorate treating bioreactor.

    PubMed

    Anupama, V N; Prajeesh, P V G; Anju, S; Priya, P; Krishnakumar, B

    2015-08-01

    A microbial consortium reducing high level of perchlorate was developed and in a fed batch bioreactor using acetate as substrate perchlorate was reduced at 0.25 g/g vss. day. Under stable performance, the microbial community structure of the reactor was analyzed through molecular and phenotypic methods. The diversity of bacteria and archaea were analyzed through whole cell Fluorescence In-Situ Hybridization (FISH) and PCR-Denaturing Gradient Gel Electrophoresis (DGGE), whereas higher trophic community was analyzed phenotypically. FISH analysis revealed the presence of alpha, beta, gamma and delta proteobacteria in the sludge, dominated by beta proteobacteria (68.7%). DGGE analysis of bacteria revealed the presence of a single known perchlorate reducing bacterium-Dechloromonas, nitrate reducers like Thaeura and Azoarcus and a number of other genera so far not reported as perchlorate or nitrate reducing. The archaea community was represented by an acetoclastic methanogen, Methanosaeta harundinacea. We have also observed the presence of an acetate consuming flagellate, Polytomella sp. in significant number in the reactor. Archaea and protozoa community in perchlorate treating bioreactor is reported first time in this study and point out further the significance of non perchlorate reducing but acetate scavenging microbial groups in acetate fed perchlorate treating reactors.

  1. Packing developments improve valve availability

    SciTech Connect

    Aikin, J.

    1994-01-01

    Regulatory environment and clean air legislation demand improvements in valve stuffing box performance. In normally inaccessible or hazardous areas, the cost of valve maintenance and repair in terms of safety, radiation, and toxic gas exposure is very high. AECL Research`s Mechanical Equipment Development (MED) branch at Chalk River Laboratories (Chalk River, Ont.) has researched stem packing leakage problems since the early 1970s. Early research and development (R and D) significantly improved the understanding of operational characteristics of asbestos-based valve packing. Of note was the development of live loading, a stem packing method that reduces valve leakage to near zero under most operating conditions, and significantly prolongs packing life. This article describes how stem packing developments, including live loading techniques, are reducing valve fugitive emissions and leakage.

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

    PubMed

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

    2015-11-01

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

  3. Establishment and development of ruminal hydrogenotrophs in methanogen-free lambs.

    PubMed

    Fonty, Gérard; Joblin, Keith; Chavarot, Michel; Roux, Remy; Naylor, Graham; Michallon, Fabien

    2007-10-01

    The aim of this work was to determine whether reductive acetogenesis can provide an alternative to methanogenesis in the rumen. Gnotobiotic lambs were inoculated with a functional rumen microbiota lacking methanogens and reared to maturity on a fibrous diet. Lambs with a methanogen-free rumen grew well, and the feed intake and ruminal volatile fatty acid concentrations for lambs lacking ruminal methanogens were lower but not markedly dissimilar from those for conventional lambs reared on the same diet. A high population density (10(7) to 10(8) cells g(-1)) of ruminal acetogens slowly developed in methanogen-free lambs. Sulfate- and fumarate-reducing bacteria were present, but their population densities were highly variable. In methanogen-free lambs, the hydrogen capture from fermentation was low (28 to 46%) in comparison with that in lambs containing ruminal methanogens (>90%). Reductive acetogenesis was not a significant part of ruminal fermentation in conventional lambs but contributed 21 to 25% to the fermentation in methanogen-free meroxenic animals. Ruminal H(2) utilization was lower in lambs lacking ruminal methanogens, but when a methanogen-free lamb was inoculated with a methanogen, the ruminal H(2) utilization was similar to that in conventional lambs. H(2) utilization in lambs containing a normal ruminal microflora was age dependent and increased with the animal age. The animal age effect was less marked in lambs lacking ruminal methanogens. Addition of fumarate to rumen contents from methanogen-free lambs increased H(2) utilization. These findings provide the first evidence from animal studies that reductive acetogens can sustain a functional rumen and replace methanogens as a sink for H(2) in the rumen.

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

    PubMed

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

    2016-01-01

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

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

    PubMed Central

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

    2016-01-01

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

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

    NASA Technical Reports Server (NTRS)

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

    1988-01-01

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

  7. Nickel requirement and factor F430 content of methanogenic bacteria.

    PubMed Central

    Diekert, G; Konheiser, U; Piechulla, K; Thauer, R K

    1981-01-01

    Methanobacterium thermoautotrophicum has been reported to require nickel for growth and to contain high concentrations of a nickel tetrapyrrole designated factor F430. In this communication it is shown that all methanogenic bacteria investigated incorporated nickel during growth and also synthesized factor F430. This was also true for Methanobrevibacter smithii, which is dependent on acetate as a carbon source, and for Methanosarcina barkeri growing on acetate or methanol as energy sources. Other bacteria, including Acetobacterium woodii and Clostridium thermoaceticum, contained no factor F430. It is further shown that two yellow nickel-containing degradation products were formed from factor F430 when heated at pH 7. This finding explains why several forms of factor F430 were found in methanogenic bacteria when a heat step was employed in the purification procedure. PMID:7298577

  8. Composition and Role of Extracellular Polymers in Methanogenic Granules

    PubMed Central

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

    1997-01-01

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

  9. 7 CFR 51.310 - Packing requirements.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... STANDARDS) United States Standards for Grades of Apples Packing Requirements § 51.310 Packing requirements. (a) Apples tray packed or cell packed in cartons shall be arranged according to approved and... that apples are of the proper size for molds or cell compartments in which they are packed, and...

  10. 7 CFR 51.310 - Packing requirements.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... STANDARDS) United States Standards for Grades of Apples Packing Requirements § 51.310 Packing requirements. (a) Apples tray packed or cell packed in cartons shall be arranged according to approved and... that apples are of the proper size for molds or cell compartments in which they are packed, and...

  11. 7 CFR 51.310 - Packing requirements.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... STANDARDS) United States Standards for Grades of Apples Packing Requirements § 51.310 Packing requirements. (a) Apples tray packed or cell packed in cartons shall be arranged according to approved and... that apples are of the proper size for molds or cell compartments in which they are packed, and...

  12. Tissue grown in space in NASA Bioreactor

    NASA Technical Reports Server (NTRS)

    1998-01-01

    For 5 days on the STS-70 mission, a bioreactor cultivated human colon cancer cells, such as the culture section shown here, which grew to 30 times the volume of control specimens grown on Earth. This significant result was reproduced on STS-85 which grew mature structures that more closely match what are found in tumors in humans. The two white circles within the tumor are part of a plastic lattice that helped the cells associate. The work is sponsored by NASA's Office of Biological and Physical Research. The bioreactor is managed by the Biotechnology Cell Science Program at NASA's Johnson Space Center (JSC). The NASA Bioreactor provides a low turbulence culture environment which promotes the formation of large, three-dimensional cell clusters. Due to their high level of cellular organization and specialization, samples constructed in the bioreactor more closely resemble the original tumor or tissue found in the body. NASA-sponsored bioreactor research has been instrumental in helping scientists to better understand normal and cancerous tissue development. In cooperation with the medical community, the bioreactor design is being used to prepare better models of human colon, prostate, breast and ovarian tumors. Cartilage, bone marrow, heart muscle, skeletal muscle, pancreatic islet cells, liver and kidney are just a few of the normal tissues being cultured in rotating bioreactors by investigators.

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

    PubMed Central

    Susanti, Dwi; Mukhopadhyay, Biswarup

    2012-01-01

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

  14. Transformation of phenol into phenylalanine by a methanogenic consortium

    SciTech Connect

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

    1996-03-01

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

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

    PubMed

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

    2004-09-28

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

  16. Environmental selection of planktonic methanogens in permafrost thaw ponds

    PubMed Central

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

    2016-01-01

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

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

    PubMed

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

    2016-01-01

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

  18. Genetics and molecular biology of methanogen genes. Final report

    SciTech Connect

    Konisky, J.

    1997-10-07

    Adenylate kinase has been isolated from four related methanogenic members of the Archaea. For each the optimum temperature for enzyme activity was similar to the temperature for optimal microbial growth and was approximately 30 C for Methanococcus voltage, 70 C for Methanococcus thermolithotrophicus, 80 C for Methanococcus igneus and 80--90 C for Methanococcus jannaschii. The enzymes were sensitive to the adenylate kinase inhibitor, Ap{sub 5}A [P{sup 1}, P{sup 5}-di(adenosine-5{prime}) pentaphosphate], a property that was exploited to purify the enzymes by CIBACRON Blue affinity chromatography. The enzymes had an estimated molecular weight (approximately 23--25 kDa) in the range common for adenylate kinases. Each of the enzymes had a region of amino acid sequence close to its N-terminus that was similar to the canonical P-loop sequence reported for all adenylate kinases. However, the methanogen sequences lacked a lysine residue that has previously been found to be invariant in adenylate kinases including an enzyme isolated from the Archeon, Sulfolobus acidocaldarius. If verified as a nucleotide binding domain, the methanogen sequence would represent a novel nucleotide binding motif. There was no correlation between amino acid abundance and the optimal temperature for enzyme activity.

  19. Hydrogen consumption by methanogens on the early Earth

    NASA Technical Reports Server (NTRS)

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

    1998-01-01

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

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

    PubMed

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

    2013-02-01

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

  1. Environmental selection of planktonic methanogens in permafrost thaw ponds

    NASA Astrophysics Data System (ADS)

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

    2016-08-01

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

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

    PubMed

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

    2016-01-01

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

  3. Role of nickel in high rate methanol degradation in anaerobic granular sludge bioreactors

    PubMed Central

    Fermoso, Fernando G.; Collins, Gavin; Bartacek, Jan; O’Flaherty, Vincent

    2008-01-01

    The effect of nickel deprivation from the influent of a mesophilic (30°C) methanol fed upflow anaerobic sludge bed (UASB) reactor was investigated by coupling the reactor performance to the evolution of the Methanosarcina population of the bioreactor sludge. The reactor was operated at pH 7.0 and an organic loading rate (OLR) of 5–15 g COD l−1 day−1 for 191 days. A clear limitation of the specific methanogenic activity (SMA) on methanol due to the absence of nickel was observed after 129 days of bioreactor operation: the SMA of the sludge in medium with the complete trace metal solution except nickel amounted to 1.164 (±0.167) g CH4-COD g VSS−1 day−1 compared to 2.027 (±0.111) g CH4-COD g VSS−1 day−1 in a medium with the complete (including nickel) trace metal solution. The methanol removal efficiency during these 129 days was 99%, no volatile fatty acid (VFA) accumulation was observed and the size of the Methanosarcina population increased compared to the seed sludge. Continuation of the UASB reactor operation with the nickel limited sludge lead to incomplete methanol removal, and thus methanol accumulation in the reactor effluent from day 142 onwards. This methanol accumulation subsequently induced an increase of the acetogenic activity in the UASB reactor on day 160. On day 165, 77% of the methanol fed to the system was converted to acetate and the Methanosarcina population size had substantially decreased. Inclusion of 0.5 μM Ni (dosed as NiCl2) to the influent from day 165 onwards lead to the recovery of the methanol removal efficiency to 99% without VFA accumulation within 2 days of bioreactor operation. PMID:18247139

  4. Spatial Experiment Technologies Suitable for Unreturnable Bioreactor

    NASA Astrophysics Data System (ADS)

    Zhang, Tao; Zheng, Weibo; Tong, Guanghui

    2016-07-01

    The system composition and main function of the bioreactor piggybacked on TZ cargo transport spacecraft are introduced briefly in the paper.The spatial experiment technologies which are suitable for unreturnable bioreactor are described in detail,including multi-channel liquid transportion and management,multi-type animal cells circuit testing,dynamic targets microscopic observation in situ etc..The feasibility and effectiveness of these technologies which will be used in space experiment in bioreactor are verified in tests and experiments on the ground.

  5. Development of Fundamental Technologies for Micro Bioreactors

    NASA Astrophysics Data System (ADS)

    Sato, Kiichi; Kitamori, Takehiko

    This chapter reviews the development of fundamental technologies required for microchip-based bioreactors utilizing living mammalian cells and pressure driven flow. The most important factor in the bioreactor is the cell culture. For proper cell culturing, continuous medium supply from a microfluidic channel and appropriate modification of the channel surface to accommodate cell attachment is required. Moreover, the medium flow rate should be chosen carefully, because shear stress affects cell activity. The techniques presented here could be applied to the development of micro bioreactors such as microlivers, pigment production by plant cells, and artificial insemination.

  6. Tubular membrane bioreactors for biotechnological processes.

    PubMed

    Wolff, Christoph; Beutel, Sascha; Scheper, Thomas

    2013-02-01

    This article is an overview of bioreactors using tubular membranes such as hollow fibers or ceramic capillaries for cultivation processes. This diverse group of bioreactor is described here in regard to the membrane materials used, operational modes, and configurations. The typical advantages of this kind of system such as environments with low shear stress together with high cell densities and also disadvantages like poor oxygen supply are summed up. As the usage of tubular membrane bioreactors is not restricted to a certain organism, a brief overview of various applications covering nearly all types of cells from prokaryotic to eukaryotic cells is also given here. PMID:23224587

  7. Bioreactor Technology in Cardiovascular Tissue Engineering

    NASA Astrophysics Data System (ADS)

    Mertsching, H.; Hansmann, J.

    Cardiovascular tissue engineering is a fast evolving field of biomedical science and technology to manufacture viable blood vessels, heart valves, myocar-dial substitutes and vascularised complex tissues. In consideration of the specific role of the haemodynamics of human circulation, bioreactors are a fundamental of this field. The development of perfusion bioreactor technology is a consequence of successes in extracorporeal circulation techniques, to provide an in vitro environment mimicking in vivo conditions. The bioreactor system should enable an automatic hydrodynamic regime control. Furthermore, the systematic studies regarding the cellular responses to various mechanical and biochemical cues guarantee the viability, bio-monitoring, testing, storage and transportation of the growing tissue.

  8. Valve stem and packing assembly

    DOEpatents

    Wordin, John J.

    1991-01-01

    A valve stem and packing assembly is provided in which a rotatable valve stem includes a first tractrix surface for sliding contact with a stem packing and also includes a second tractrix surface for sliding contact with a bonnet. Force is applied by means of a spring, gland flange, and gland on the stem packing so the stem packing seals to the valve stem and bonnet. This configuration serves to create and maintain a reliable seal between the stem packing and the valve stem. The bonnet includes a second complementary tractrix surface for contacting the second sliding tractrix surface, the combination serving as a journal bearing for the entire valve stem and packing assembly. The journal bearing so configured is known as a Schiele's pivot. The Schiele's pivot also serves to maintain proper alignment of the valve stem with respect to the bonnet. Vertical wear between the surfaces of the Schiele's pivot is uniform at all points of contact between the second sliding tractrix surface and the second complementary tractrix surface of a bonnet. The valve stem is connected to a valve plug by means of a slip joint. The valve is opened and closed by rotating the valve stem. The slip joint compensates for wear on the Schiele's pivot and on the valve plug. A ledge is provided on the valve bonnet for the retaining nut to bear against. The ledge prevents overtightening of the retaining nut and the resulting excessive friction between stem and stem packing.

  9. Valve stem and packing assembly

    DOEpatents

    Wordin, J.J.

    1991-09-03

    A valve stem and packing assembly is provided in which a rotatable valve stem includes a first tractrix surface for sliding contact with a stem packing and also includes a second tractrix surface for sliding contact with a bonnet. Force is applied by means of a spring, gland flange, and gland on the stem packing so the stem packing seals to the valve stem and bonnet. This configuration serves to create and maintain a reliable seal between the stem packing and the valve stem. The bonnet includes a second complementary tractrix surface for contacting the second sliding tractrix surface, the combination serving as a journal bearing for the entire valve stem and packing assembly. The journal bearing so configured is known as a Schiele's pivot. The Schiele's pivot also serves to maintain proper alignment of the valve stem with respect to the bonnet. Vertical wear between the surfaces of the Schiele's pivot is uniform at all points of contact between the second sliding tractrix surface and the second complementary tractrix surface of a bonnet. The valve stem is connected to a valve plug by means of a slip joint. The valve is opened and closed by rotating the valve stem. The slip joint compensates for wear on the Schiele's pivot and on the valve plug. A ledge is provided on the valve bonnet for the retaining nut to bear against. The ledge prevents over tightening of the retaining nut and the resulting excessive friction between stem and stem packing. 2 figures.

  10. Continuous anaerobic bioreactor with a fixed-structure bed (ABFSB) for wastewater treatment with low solids and low applied organic loading content.

    PubMed

    Mockaitis, G; Pantoja, J L R; Rodrigues, J A D; Foresti, E; Zaiat, M

    2014-07-01

    This paper describes a new type of anaerobic bioreactor with a fixed-structure bed (ABFSB) in which the support for the biomass consists of polyurethane foam strips placed along the length of the bioreactor. This configuration prevents the accumulation of biomass or solids in the bed as well as clogging and channeling effects. In this study, complex synthetic wastewater with a chemical oxygen demand of 404.4 mg O(2) L(-1) is treated by the reactor. The ABFSB, which has a working volume of 4.77 L, was inoculated with anaerobic sludge obtained from an upflow anaerobic sludge blanket bioreactor. A removal efficiency of 78 % for organic matter and an effluent pH of 6.97 were achieved. An analysis of the organic volatile acids produced by the ABFSB indicated that it operated under stable conditions during an experimental run of 36 days. The stable and efficient operation of the bioreactor was compared with the configurations of other anaerobic bioreactors used for complex wastewater treatment. The results of the study indicate that the ABFSB is a technological alternative to packed-bed bioreactors.

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

    PubMed Central

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

    2015-01-01

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

  12. Identification of Methanogenic archaea in the Hyporheic Sediment of Sitka Stream

    PubMed Central

    Buriánková, Iva; Brablcová, Lenka; Mach, Václav; Dvořák, Petr; Chaudhary, Prem Prashant; Rulík, Martin

    2013-01-01

    Methanogenic archaea produce methane as a metabolic product under anoxic conditions and they play a crucial role in the global methane cycle. In this study molecular diversity of methanogenic archaea in the hyporheic sediment of the lowland stream Sitka (Olomouc, Czech Republic) was analyzed by PCR amplification, cloning and sequencing analysis of the methyl coenzyme M reductase alpha subunit (mcrA) gene. Sequencing analysis of 60 clones revealed 24 different mcrA phylotypes from hyporheic sedimentary layers to a depth of 50 cm. Phylotypes were affiliated with Methanomicrobiales, Methanosarcinales and Methanobacteriales orders. Only one phylotype remains unclassified. The majority of the phylotypes showed higher affiliation with uncultured methanogens than with known methanogenic species. The presence of relatively rich assemblage of methanogenic archaea confirmed that methanogens may be an important component of hyporheic microbial communities and may affect CH4 cycling in rivers. PMID:24278322

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

    PubMed

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

    2015-01-01

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

  14. 7 CFR 982.11 - Pack.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... the Department of Agriculture (Continued) AGRICULTURAL MARKETING SERVICE (Marketing Agreements and... Order Regulating Handling Definitions § 982.11 Pack. Pack means a specific commercial classification according to size, internal quality, and external appearance and condition of hazelnuts packed in...

  15. Energetic and hydrogen limitations of thermophilic and hyperthermophilic methanogens

    NASA Astrophysics Data System (ADS)

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

    2013-12-01

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

  16. Method for packing chromatographic beds

    DOEpatents

    Freeman, David H.; Angeles, Rosalie M.; Keller, Suzanne

    1991-01-01

    Column chromatography beds are packed through the application of static force. A slurry of the chromatography bed material and a non-viscous liquid is filled into the column plugged at one end, and allowed to settle. The column is transferred to a centrifuge, and centrifuged for a brief period of time to achieve a predetermined packing level, at a range generally of 100-5,000 gravities. Thereafter, the plug is removed, other fixtures may be secured, and the liquid is allowed to flow out through the bed. This results in an evenly packed bed, with no channeling or preferential flow characteristics.

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

    PubMed

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

    2016-09-01

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

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

    PubMed

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

    2016-09-01

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

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

    PubMed

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

    2015-06-30

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

  20. Anaerobic biodegradation of 2,4,6-trichlorophenol by methanogenic granular sludge: role of co-substrates and methanogenic inhibition.

    PubMed

    Puyol, D; Mohedano, A F; Sanz, J L; Rodríguez, J J

    2009-01-01

    The influence of several co-substrates in the anaerobic biodegradation of 2,4,6-trichlorophenol (246TCP) by methanogenic granular sludge as well as in methanogenesis inhibition by 246TCP has been studied. 4 g-COD.L(-1) of lactate, sucrose, volatile fatty acids (VFA) acetate:propionate:butyrate 1:1:1, ethanol, methanol, yeast extract (YE), and 2 g-COD.L(-1) of formate and methylamine were tested. Two concentrations of 246TCP: 80 mg.L(-1) and 113 mg.L(-1) (this last corresponding to the EC(50) for acetotrophic methanogenesis) were tested. Three consecutive co-substrate and nutrient feedings were accomplished. 246TCP was added in the second feed, and the 246TCP removal rate increased considerably after the third feed. Accumulated metabolites after ortho-dechlorination, either 4-chlorophenol (4CP) (when methanol, ethanol or VFA were used as co-substrates) or 2,4-dichlorophenol (24DCP) (with lactate) avoided the complete dechlorination of 246TCP. With methylamine and formate this compound was degraded only partially. Monochlorophenols biodegradation was partially achieved with YE, but both 24DCP and 2,6-dichlorophenol (26DCP) were accumulated. In the presence of sucrose para-dechlorination was observed. 246TCP was better tolerated by methanogens when ethanol and methanol were added because of the highest specific methanogenic activity achieved with these co-substrates. Methanol and ethanol were the best co-substrates in the anaerobic biodegradation of 246TCP.

  1. An innovative membrane bioreactor for methane biohydroxylation.

    PubMed

    Pen, N; Soussan, L; Belleville, M-P; Sanchez, J; Charmette, C; Paolucci-Jeanjean, D

    2014-12-01

    In this study, a membrane bioreactor (MBR) was developed for efficient, safe microbial methane hydroxylation with Methylosinus trichosporium OB3b. This innovative MBR, which couples a bioreactor with two gas/liquid macroporous membrane contactors supplying the two gaseous substrates (methane and oxygen) was operated in fed-batch mode. The feasibility and the reproducibility of this new biohydroxylation process were first demonstrated. The mass transfer within this MBR was twice that observed in a batch reactor in similar conditions. The productivity reached with this MBR was 75±25mgmethanol(gdrycell)(-1)h(-1). Compared to the literature, this value is 35times higher than that obtained with the only other fed-batch membrane bioreactor reported, which was run with dense membranes, and is comparable to those obtained with bioreactors fed by bubble-spargers. However, in the latter case, an explosive gas mixture can be formed, a problem that is avoided with the MBR.

  2. In vivo bioreactors for mandibular reconstruction.

    PubMed

    Tatara, A M; Wong, M E; Mikos, A G

    2014-12-01

    Large mandibular defects are difficult to reconstruct with good functional and aesthetic outcomes because of the complex geometry of craniofacial bone. While the current gold standard is free tissue flap transfer, this treatment is limited in fidelity by the shape of the harvested tissue and can result in significant donor site morbidity. To address these problems, in vivo bioreactors have been explored as an approach to generate autologous prefabricated tissue flaps. These bioreactors are implanted in an ectopic site in the body, where ossified tissue grows into the bioreactor in predefined geometries and local vessels are recruited to vascularize the developing construct. The prefabricated flap can then be harvested with vessels and transferred to a mandibular defect for optimal reconstruction. The objective of this review article is to introduce the concept of the in vivo bioreactor, describe important preclinical models in the field, summarize the human cases that have been reported through this strategy, and offer future directions for this exciting approach.

  3. Bioreactor Design for Tendon/Ligament Engineering

    PubMed Central

    Wang, Tao; Gardiner, Bruce S.; Lin, Zhen; Rubenson, Jonas; Kirk, Thomas B.; Wang, Allan; Xu, Jiake

    2013-01-01

    Tendon and ligament injury is a worldwide health problem, but the treatment options remain limited. Tendon and ligament engineering might provide an alternative tissue source for the surgical replacement of injured tendon. A bioreactor provides a controllable environment enabling the systematic study of specific biological, biochemical, and biomechanical requirements to design and manufacture engineered tendon/ligament tissue. Furthermore, the tendon/ligament bioreactor system can provide a suitable culture environment, which mimics the dynamics of the in vivo environment for tendon/ligament maturation. For clinical settings, bioreactors also have the advantages of less-contamination risk, high reproducibility of cell propagation by minimizing manual operation, and a consistent end product. In this review, we identify the key components, design preferences, and criteria that are required for the development of an ideal bioreactor for engineering tendons and ligaments. PMID:23072472

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

    PubMed

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

    2015-01-01

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

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

    PubMed Central

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

    2015-01-01

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

  6. Anaerobic microbial community response to methanogenic inhibitors 2-bromoethanesulfonate and propynoic acid.

    PubMed

    Webster, Tara M; Smith, Adam L; Reddy, Raghav R; Pinto, Ameet J; Hayes, Kim F; Raskin, Lutgarde

    2016-08-01

    Methanogenic inhibitors are often used to study methanogenesis in complex microbial communities or inhibit methanogens in the gastrointestinal tract of livestock. However, the resulting structural and functional changes in archaeal and bacterial communities are poorly understood. We characterized microbial community structure and activity in mesocosms seeded with cow dung and municipal wastewater treatment plant anaerobic digester sludge after exposure to two methanogenic inhibitors, 2-bromoethanesulfonate (BES) and propynoic acid (PA). Methane production was reduced by 89% (0.5 mmol/L BES), 100% (10 mmol/LBES), 24% (0.1 mmol/LPA), and 95% (10 mmol/LPA). Using modified primers targeting the methyl-coenzyme M reductase (mcrA) gene, changes in mcrA gene expression were found to correspond with changes in methane production and the relative activity of methanogens. Methanogenic activity was determined by the relative abundance of methanogen 16S rRNA cDNA as a percentage of the total community 16S rRNA cDNA. Overall, methanogenic activity was lower when mesocosms were exposed to higher concentrations of both inhibitors, and aceticlastic methanogens were inhibited to a greater extent than hydrogenotrophic methanogens. Syntrophic bacterial activity, measured by 16S rRNA cDNA, was also reduced following exposure to both inhibitors, but the overall structure of the active bacterial community was not significantly affected. PMID:26987552

  7. Methane production and methanogenic Archaea in the digestive tracts of millipedes (Diplopoda).

    PubMed

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

    2014-01-01

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

  8. Thin film bioreactors in space.

    PubMed

    Hughes-Fulford, M; Scheld, H W

    1989-01-01

    Studies from the Skylab, SL-3 and D-1 missions have demonstrated that biological organisms grown in microgravity have changes in basic cellular functions such as DNA, mRNA and protein synthesis, cytoskeleton synthesis, glucose utilization and cellular differentiation. Since microgravity could affect prokaryotic and eukaryotic cells at a subcellular and molecular level, space offers us an opportunity to learn more about basic biological systems with one important variable removed. The thin film bioreactor will facilitate the handling of fluids in microgravity, under constant temperature and will allow multiple samples of cells to be grown with variable conditions. Studies on cell cultures grown in microgravity would enable us to identify and quantify changes in basic biological function in microgravity which are needed to develop new applications of orbital research and future biotechnology.

  9. Thin film bioreactors in space

    NASA Technical Reports Server (NTRS)

    Hughes-Fulford, M.; Scheld, H. W.

    1989-01-01

    Studies from the Skylab, SL-3 and D-1 missions have demonstrated that biological organisms grown in microgravity have changes in basic cellular functions such as DNA, mRNA and protein synthesis, cytoskeleton synthesis, glucose utilization, and cellular differentiation. Since microgravity could affect prokaryotic and eukaryotic cells at a subcellular and molecular level, space offers an opportunity to learn more about basic biological systems with one inmportant variable removed. The thin film bioreactor will facilitate the handling of fluids in microgravity, under constant temperature and will allow multiple samples of cells to be grown with variable conditions. Studies on cell cultures grown in microgravity would make it possible to identify and quantify changes in basic biological function in microgravity which are needed to develop new applications of orbital research and future biotechnology.

  10. NASA Bioreactors Advance Disease Treatments

    NASA Technical Reports Server (NTRS)

    2009-01-01

    The International Space Station (ISS) is falling. This is no threat to the astronauts onboard, however, because falling is part of the ISS staying in orbit. The absence of gravity beyond the Earth s atmosphere is actually an illusion; at the ISS s orbital altitude of approximately 250 miles above the surface, the planet s gravitational pull is only 12-percent weaker than on the ground. Gravity is constantly pulling the ISS back to Earth, but the space station is also constantly traveling at nearly 18,000 miles per hour. This means that, even though the ISS is falling toward Earth, it is moving sideways fast enough to continually miss impacting the planet. The balance between the force of gravity and the ISS s motion creates a stable orbit, and the fact that the ISS and everything in it including the astronauts are falling at an equal rate creates the condition of weightlessness called microgravity. The constant falling of objects in orbit is not only an important principle in space, but it is also a key element of a revolutionary NASA technology here on Earth that may soon help cure medical ailments from heart disease to diabetes. In the mid-1980s, NASA researchers at Johnson Space Center were investigating the effects of long-term microgravity on human tissues. At the time, the Agency s shuttle fleet was grounded following the 1986 Space Shuttle Challenger disaster, and researchers had no access to the microgravity conditions of space. To provide a method for recreating such conditions on Earth, Johnson s David Wolf, Tinh Trinh, and Ray Schwarz developed that same year a horizontal, rotating device called a rotating wall bioreactor that allowed the growth of human cells in simulated weightlessness. Previously, cell cultures on Earth could only be grown two-dimensionally in Petri dishes, because gravity would cause the multiplying cells to sink within their growth medium. These cells do not look or function like real human cells, which grow three-dimensionally in

  11. Energy efficiency in membrane bioreactors.

    PubMed

    Barillon, B; Martin Ruel, S; Langlais, C; Lazarova, V

    2013-01-01

    Energy consumption remains the key factor for the optimisation of the performance of membrane bioreactors (MBRs). This paper presents the results of the detailed energy audits of six full-scale MBRs operated by Suez Environnement in France, Spain and the USA based on on-site energy measurement and analysis of plant operation parameters and treatment performance. Specific energy consumption is compared for two different MBR configurations (flat sheet and hollow fibre membranes) and for plants with different design, loads and operation parameters. The aim of this project was to understand how the energy is consumed in MBR facilities and under which operating conditions, in order to finally provide guidelines and recommended practices for optimisation of MBR operation and design to reduce energy consumption and environmental impacts.

  12. Simplified Bioreactor For Growing Mammalian Cells

    NASA Technical Reports Server (NTRS)

    Spaulding, Glenn F.

    1995-01-01

    Improved bioreactor for growing mammalian cell cultures developed. Designed to support growth of dense volumes of mammalian cells by providing ample, well-distributed flows of nutrient solution with minimal turbulence. Cells relatively delicate and, unlike bacteria, cannot withstand shear forces present in turbulent flows. Bioreactor vessel readily made in larger sizes to accommodate greater cell production quantities. Molding equipment presently used makes cylinders up to 30 centimeters long. Alternative sintered plastic techniques used to vary pore size and quantity, as necessary.

  13. Colon tumor cells grown in NASA Bioreactor

    NASA Technical Reports Server (NTRS)

    2001-01-01

    These photos compare the results of colon carcinoma cells grown in a NASA Bioreactor flown on the STS-70 Space Shuttle in 1995 flight and ground control experiments. The cells grown in microgravity (left) have aggregated to form masses that are larger and more similar to tissue found in the body than the cells cultured on the ground (right). The principal investigator is Milburn Jessup of the University of Texas M. D. Anderson Cancer Center. The NASA Bioreactor provides a low turbulence culture environment which promotes the formation of large, three-dimensional cell clusters. Due to their high level of cellular organization and specialization, samples constructed in the bioreactor more closely resemble the original tumor or tissue found in the body. NASA-sponsored bioreactor research has been instrumental in helping scientists to better understand normal and cancerous tissue development. In cooperation with the medical community, the bioreactor design is being used to prepare better models of human colon, prostate, breast and ovarian tumors. Cartilage, bone marrow, heart muscle, skeletal muscle, pancreatic islet cells, liver and kidney are just a few of the normal tissues being cultured in rotating bioreactors by investigators. Cell constructs grown in a rotating bioreactor on Earth (left) eventually become too large to stay suspended in the nutrient media. In the microgravity of orbit, the cells stay suspended. Rotation then is needed for gentle stirring to replenish the media around the cells. The work is sponsored by NASA's Office of Biological and Physical Research. The bioreactor is managed by the Biotechnology Cell Science Program at NASA's Johnson Space Center (JSC). Credit: NASA and University of Texas M. D. Anderson Cancer Center.

  14. Heart tissue grown in NASA Bioreactor

    NASA Technical Reports Server (NTRS)

    2001-01-01

    Lisa Freed and Gordana Vunjak-Novakovic, both of the Massachusetts Institute of Technology (MIT), have taken the first steps toward engineering heart muscle tissue that could one day be used to patch damaged human hearts. Cells isolated from very young animals are attached to a three-dimensional polymer scaffold, then placed in a NASA bioreactor. The cells do not divide, but after about a week start to cornect to form a functional piece of tissue. Functionally connected heart cells that are capable of transmitting electrical signals are the goal for Freed and Vunjak-Novakovic. Electrophysiological recordings of engineered tissue show spontaneous contractions at a rate of 70 beats per minute (a), and paced contractions at rates of 80, 150, and 200 beats per minute respectively (b, c, and d). The NASA Bioreactor provides a low turbulence culture environment which promotes the formation of large, three-dimensional cell clusters. The Bioreactor is rotated to provide gentle mixing of fresh and spent nutrient without inducing shear forces that would damage the cells. Due to their high level of cellular organization and specialization, samples constructed in the bioreactor more closely resemble the original tumor or tissue found in the body. NASA-sponsored bioreactor research has been instrumental in helping scientists to better understand normal and cancerous tissue development. In cooperation with the medical community, the bioreactor design is being used to prepare better models of human colon, prostate, breast and ovarian tumors. Cartilage, bone marrow, heart muscle, skeletal muscle, pancreatic islet cells, liver and kidney are just a few of the normal tissues being cultured in rotating bioreactors by investigators. The work is sponsored by NASA's Office of Biological and Physical Research. The bioreactor is managed by the Biotechnology Cell Science Program at NASA's Johnson Space Center (JSC). Credit: NASA and MIT.

  15. Heart tissue grown in NASA Bioreactor

    NASA Technical Reports Server (NTRS)

    2001-01-01

    Lisa Freed and Gordana Vunjak-Novakovic, both of the Massachusetts Institute of Technology (MIT), have taken the first steps toward engineering heart muscle tissue that could one day be used to patch damaged human hearts. Cells isolated from very young animals are attached to a three-dimensional polymer scaffold, then placed in a NASA bioreactor. The cells do not divide, but after about a week start to cornect to form a functional piece of tissue. Here, a transmission electron micrograph of engineered tissue shows a number of important landmarks present in functional heart tissue: (A) well-organized myofilaments (Mfl), z-lines (Z), and abundant glycogen granules (Gly); and (D) intercalcated disc (ID) and desmosomes (DES). The NASA Bioreactor provides a low turbulence culture environment which promotes the formation of large, three-dimensional cell clusters. The Bioreactor is rotated to provide gentle mixing of fresh and spent nutrient without inducing shear forces that would damage the cells. Due to their high level of cellular organization and specialization, samples constructed in the bioreactor more closely resemble the original tumor or tissue found in the body. NASA-sponsored bioreactor research has been instrumental in helping scientists to better understand normal and cancerous tissue development. In cooperation with the medical community, the bioreactor design is being used to prepare better models of human colon, prostate, breast and ovarian tumors. Cartilage, bone marrow, heart muscle, skeletal muscle, pancreatic islet cells, liver and kidney are just a few of the normal tissues being cultured in rotating bioreactors by investigators. The work is sponsored by NASA's Office of Biological and Physical Research. The bioreactor is managed by the Biotechnology Cell Science Program at NASA's Johnson Space Center (JSC). Credit: MIT

  16. 2,4-Dichlorophenol (DCP) containing wastewater treatment using a hybrid-loop bioreactor.

    PubMed

    Dilaver, Mehmet; Kargi, Fikret

    2009-02-01

    Synthetic wastewater containing 2,4-dichlorophenol (DCP) was biologically treated using a hybrid-loop bioreactor system consisting of a packed column biofilm reactor (PCBR) and an aerated tank with effluent recycle. Effects of the feed DCP concentration on COD, DCP and toxicity removals were investigated. Biomass concentration in the packed column and in the aeration tank decreased with increasing feed DCP content due to toxic effects of DCP on the microorganisms. Low biomass concentrations at high DCP contents resulted in low COD, DCP and toxicity removals. Therefore, percent DCP, COD and toxicity removals decreased with increasing feed DCP content. Nearly 70% COD removal was achieved with a feed DCP content of 380 mg L(-1). The system should be operated with the feed DCP lower than 100 mg L(-1) in order to obtain DCP, COD and toxicity removals above 90%.

  17. Are proteins well-packed?

    PubMed

    Liang, J; Dill, K A

    2001-08-01

    The average packing density inside proteins is as high as in crystalline solids. Does this mean proteins are well-packed? We go beyond average densities, and look at the full distribution functions of free volumes inside proteins. Using a new and rigorous Delaunay triangulation method for parsing space into empty and filled regions, we introduce formal definitions of interior and surface packing densities. Although proteins look like organic crystals by the criterion of average density, they look more like liquids and glasses by the criterion of their free volume distributions. The distributions are broad, and the scalings of volume-to-surface, volume-to-cluster-radius, and numbers of void versus volume show that the interiors of proteins are more like randomly packed spheres near their percolation threshold than like jigsaw puzzles. We find that larger proteins are packed more loosely than smaller proteins. And we find that the enthalpies of folding (per amino acid) are independent of the packing density of a protein, indicating that van der Waals interactions are not a dominant component of the folding forces. PMID:11463623

  18. Estimation of flow and transport parameters for woodchip based bioreactors: I. laboratory-scale bioreactor

    Technology Transfer Automated Retrieval System (TEKTRAN)

    In subsurface bioreactors used for tile drainage systems, carbon sources are used to facilitate denitrification. The objective of this study was to estimate hydraulic conductivity, effective porosity, dispersivity, and first-order decay coefficients for a laboratory-scale bioreactor with woodchips a...

  19. Advanced microscale bioreactor system: a representative scale-down model for bench-top bioreactors.

    PubMed

    Hsu, Wei-Ting; Aulakh, Rigzen P S; Traul, Donald L; Yuk, Inn H

    2012-12-01

    In recent years, several automated scale-down bioreactor systems have been developed to increase efficiency in cell culture process development. ambr™ is an automated workstation that provides individual monitoring and control of culture dissolved oxygen and pH in single-use, stirred-tank bioreactors at a working volume of 10-15 mL. To evaluate the ambr™ system, we compared the performance of four recombinant Chinese hamster ovary cell lines in a fed-batch process in parallel ambr™, 2-L bench-top bioreactors, and shake flasks. Cultures in ambr™ matched 2-L bioreactors in controlling the environment (temperature, dissolved oxygen, and pH) and in culture performance (growth, viability, glucose, lactate, Na(+), osmolality, titer, and product quality). However, cultures in shake flasks did not show comparable performance to the ambr™ and 2-L bioreactors.

  20. Experimental investigation on feasible bioreactor using mechanism of hydrogen oxidation of natural soil for detritiation system.

    PubMed

    Edao, Yuki; Iwai, Yasunori; Sato, Katsumi; Hayashi, Takumi

    2016-08-01

    A passive reactor for tritium oxidation at room temperature has been widely studied in nuclear engineering especially for a detritiation system (DS) of a tritium process facility taking possible extraordinary situation severely into consideration. We have focused on bacterial oxidation of tritium by hydrogen-oxidizing bacteria in natural soil to realize the passive oxidation reactor. The purpose of this study was to examine the feasibility of a bioreactor with hydrogen-oxidizing bacteria in soil from a point of view of engineering. The efficiency of the bioreactor was evaluated by kinetics. The bioreactor packed with natural soil shows a relative high conversion rate of tritium under the saturated moisture condition at room temperature, which is obviously superior to that of a Pt/Al2O3 catalyst generally used for tritium oxidation in the existing tritium handling facilities. The order of reaction for tritium oxidation with soil was the pseudo-first order as assessed with Michaelis-Menten kinetics model. Our engineering suggestion to increase the reaction rate is the intentional addition of hydrogen at a small concentration in the feed gas on condition that the oxidation of tritium with soil is expressed by the Michaelis-Menten kinetics model.

  1. Degradation of hydrocarbons under methanogenic conditions in different geosystems

    NASA Astrophysics Data System (ADS)

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

    2014-05-01

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

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

    SciTech Connect

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

    1995-04-01

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

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

    PubMed

    Graham, David E; White, Robert H

    2002-04-01

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

  4. 7 CFR 51.1527 - Standard pack.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... Standards for Grades of Fresh Plums and Prunes Standard Pack § 51.1527 Standard pack. (a) Packing. (1) All... plums or prunes in the top layer of any package shall be reasonably representative in quality and size of those in the remainder of the package. (3) “Well filled” means that the plums or prunes packed...

  5. 7 CFR 51.1527 - Standard pack.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... Standards for Grades of Fresh Plums and Prunes Standard Pack § 51.1527 Standard pack. (a) Packing. (1) All... plums or prunes in the top layer of any package shall be reasonably representative in quality and size of those in the remainder of the package. (3) “Well filled” means that the plums or prunes packed...

  6. 7 CFR 51.310 - Packing requirements.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... (INSPECTION, CERTIFICATION, AND STANDARDS) United States Standards for Grades of Apples Packing Requirements § 51.310 Packing requirements. (a) Apples tray packed or cell packed in cartons shall be arranged.... 3 2 “Fairly tight” means that apples are of the proper size for molds or cell compartments in...

  7. 7 CFR 51.310 - Packing requirements.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... (INSPECTION, CERTIFICATION, AND STANDARDS) United States Standards for Grades of Apples Packing Requirements § 51.310 Packing requirements. (a) Apples tray packed or cell packed in cartons shall be arranged.... 3 2 “Fairly tight” means that apples are of the proper size for molds or cell compartments in...

  8. 7 CFR 51.1217 - Standard pack.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ..., CERTIFICATION, AND STANDARDS) United States Standards for Grades of Peaches Standard Pack § 51.1217 Standard pack. (a) Each package shall be packed so that the peaches in the shown face shall be reasonably representative in size, color and quality of the contents of the package. (b) Peaches packed in U.S....

  9. 7 CFR 51.1217 - Standard pack.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... Standards for Grades of Peaches Standard Pack § 51.1217 Standard pack. (a) Each package shall be packed so that the peaches in the shown face shall be reasonably representative in size, color and quality of the contents of the package. (b) Peaches packed in U.S. Standard bushel baskets, or half-bushel baskets,...

  10. 7 CFR 51.1217 - Standard pack.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... Standards for Grades of Peaches Standard Pack § 51.1217 Standard pack. (a) Each package shall be packed so that the peaches in the shown face shall be reasonably representative in size, color and quality of the contents of the package. (b) Peaches packed in U.S. Standard bushel baskets, or half-bushel baskets,...

  11. 7 CFR 51.1217 - Standard pack.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... Standards for Grades of Peaches Standard Pack § 51.1217 Standard pack. (a) Each package shall be packed so that the peaches in the shown face shall be reasonably representative in size, color and quality of the contents of the package. (b) Peaches packed in U.S. Standard bushel baskets, or half-bushel baskets,...

  12. 7 CFR 51.1217 - Standard pack.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ..., CERTIFICATION, AND STANDARDS) United States Standards for Grades of Peaches Standard Pack § 51.1217 Standard pack. (a) Each package shall be packed so that the peaches in the shown face shall be reasonably representative in size, color and quality of the contents of the package. (b) Peaches packed in U.S....

  13. Disc pack cleaning table saves computer time

    NASA Technical Reports Server (NTRS)

    Guy, J. T., Sr.

    1970-01-01

    Disc pack holding table is support frame upon which computer disc pack is loaded and protective cover released. This combination permits manual off-line cleaning of disc pack storage units at any time without shutting down the computer, and eliminates on-line disc drive unit to hold pack during cleaning.

  14. Nature of packs used in propellant modeling.

    PubMed

    Maggi, F; Stafford, S; Jackson, T L; Buckmaster, J

    2008-04-01

    In recent years we have constructed closely packed spheres using the Lubachevsky-Stillinger algorithm to generate morphological models of heterogeneous solid propellants. Improvements to the algorithm now allow us to create large polydisperse packs on a laptop computer, and to create monodisperse packs with packing fractions greater than 70% which display significant crystal order. The use of these models in the physical context motivates efforts to examine in some detail the nature of the packs, including certain statistical properties. We compare packing fractions for binary packs with long-known experimental data. Also, we discuss the near-neighbor number and the radial distribution function (RDF) for monodisperse packs and make comparisons with experimental data. We also briefly discuss the RDF for bidisperse packs. We also consider bounded monodisperse packs, and pay particular attention to the near-wall structure where we identify significant order.

  15. Valve packings conquer fugitive emissions

    SciTech Connect

    1995-11-01

    In the early 1990s, when the US Environmental Protection Agency (EPA; Washington, D.C.) declared its intent to regulate fugitive emissions from valve-stem leakage, much of the chemical process industries (CPI) responded with fear and uncertainty. The biggest fear was that valve packing would not meet the required limits on leak rates and that expensive bellows seals may be required on many applications. The uncertainly was about how much it would cost. Today, for the most part, these concerns have been mitigated. It is estimated that about 80--90% of valves satisfy the emission requirements. The rest need some improvement in their packing systems to meet the regulations. Generally, these valves can be brought within compliance if the packing designers follow a few basic principles: Employ less-pliable outer rings and more-pliable inner rings; and don`t use excessive packing. While interest in valve packing remains high, mechanical seals continue to become more user-friendly. Many of those covered below are designed to run dry, and some can even tolerate high shaft-wobble without damage. Also look for improved flange gaskets and a host of seals to protect bearings. Twenty-one summaries are presented on new products and services.

  16. Community Structure Analysis of Methanogens Associated with Rumen Protozoa Reveals Bias in Universal Archaeal Primers

    PubMed Central

    McAllister, Tim A.

    2012-01-01

    The diversity of protozoan-associated methanogens in cattle was investigated using five universal archaeal small-subunit (SSU) rRNA gene primer sets. Methanobrevibacter spp. and rumen cluster C (distantly related to Thermoplasma spp.) were predominant. Significant differences in species composition among libraries indicate that some primers used previously to characterize rumen methanogens exhibit biased amplification. PMID:22447586

  17. Spatial Variations of the Methanogenic Communities in the Sediments of Tropical Mangroves

    PubMed Central

    Jing, Hongmei; Cheung, Shunyan; Zhou, Zhi; Wu, Chen; Nagarajan, Sanjay; Liu, Hongbin

    2016-01-01

    Methane production by methanogens in mangrove sediments is known to contribute significantly to global warming, but studies on the shift of methanogenic community in response to anthropogenic contaminations were still limited. In this study, the effect of anthropogenic activities in the mangrove sediments along the north and south coastlines of Singapore were investigated by pyrosequencing of the mcrA gene. Our results showed that hydrogenotrophic, acetoclastic and methylotrophic methanogens coexist in the sediments. The predominance of the methylotrophic Methanosarcinales reflects the potential for high methane production as well as the possible availability of low acetate and high methylated C-1 compounds as substrates. A decline in the number of acetoclastic/methylotrophic methanogens in favor of hydrogenotrophic methanogens was observed along a vertical profile in Sungei Changi, which was contaminated by heavy metals. The diversity of methanogens in the various contaminated stations was significantly different from that in a pristine St. John’s Island. The spatial variation in the methanogenic communities among the different stations was more distinct than those along the vertical profiles at each station. We suggest that the overall heterogeneity of the methanogenic communities residing in the tropical mangrove sediments might be due to the accumulated effects of temperature and concentrations of nitrate, cobalt, and nickel. PMID:27684479

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

    PubMed Central

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

    2014-01-01

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

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

    PubMed

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

    2016-01-01

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

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

    PubMed Central

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

    2016-01-01

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

  1. Methanogenic archaea in health and disease: a novel paradigm of microbial pathogenesis.

    PubMed

    Conway de Macario, Everly; Macario, Alberto J L

    2009-02-01

    Microbes that produce methane gas, methanogens, were identified as Archaea in the 1970s but their possible role in disease is only emerging now, after they were found in the large intestine, mouth, and vagina. Significant associations were observed, for instance, between levels of methanogens in periodontal pockets and severity of periodontitis, and between quantities of methanogens in the large intestine and diseases such as colon cancer and diverticulosis. Recently, a role for intestinal methanogens in obesity was proposed. The lesson learned is that for methanogens we have to look at their pathogenicity from a different angle in comparison to classic pathogens that invade tissues and release toxins. This type of pathogenicity has not yet been described for methanogens. Instead, methanogens seem to participate in pathogenicity indirectly, favoring the growth of other microbes, which are directly involved in pathogenesis. This indirect role should not be minimized. On the contrary, it has become clear that a fundamental change of approach to the understanding and control of microbial diseases must be implemented. A comprehensive strategy is needed to elucidate the syntrophic associations that are essential for a healthy relation among microbes (including methanogens) and between them and the host organism, and to unveil those associations that lead to disease.

  2. Chasing the elusive Euryarchaeota class WSA2: genomes reveal a uniquely fastidious methyl-reducing methanogen.

    PubMed

    Nobu, Masaru Konishi; Narihiro, Takashi; Kuroda, Kyohei; Mei, Ran; Liu, Wen-Tso

    2016-10-01

    The ecophysiology of one candidate methanogen class WSA2 (or Arc I) remains largely uncharacterized, despite the long history of research on Euryarchaeota methanogenesis. To expand our understanding of methanogen diversity and evolution, we metagenomically recover eight draft genomes for four WSA2 populations. Taxonomic analyses indicate that WSA2 is a distinct class from other Euryarchaeota. None of genomes harbor pathways for CO2-reducing and aceticlastic methanogenesis, but all possess H2 and CO oxidation and energy conservation through H2-oxidizing electron confurcation and internal H2 cycling. As the only discernible methanogenic outlet, they consistently encode a methylated thiol coenzyme M methyltransferase. Although incomplete, all draft genomes point to the proposition that WSA2 is the first discovered methanogen restricted to methanogenesis through methylated thiol reduction. In addition, the genomes lack pathways for carbon fixation, nitrogen fixation and biosynthesis of many amino acids. Acetate, malonate and propionate may serve as carbon sources. Using methylated thiol reduction, WSA2 may not only bridge the carbon and sulfur cycles in eutrophic methanogenic environments, but also potentially compete with CO2-reducing methanogens and even sulfate reducers. These findings reveal a remarkably unique methanogen 'Candidatus Methanofastidiosum methylthiophilus' as the first insight into the sixth class of methanogens 'Candidatus Methanofastidiosa'.

  3. Relationship between methanogenic archaea and subgingival microbial complexes in human periodontitis.

    PubMed

    Horz, H P; Robertz, N; Vianna, M E; Henne, K; Conrads, G

    2015-10-01

    We compared the amounts of methanogenic archaea with ten of the most important periodontal pathogens in 125 clinical samples. Correlation analysis suggests that the support of the periodontitis-associated bacterial consortium by methanogenic archaea may be driven through direct or indirect interactions with Prevotella intermedia.

  4. Effect of paddy-upland rotation on methanogenic archaeal community structure in paddy field soil.

    PubMed

    Liu, Dongyan; Ishikawa, Hiroki; Nishida, Mizuhiko; Tsuchiya, Kazunari; Takahashi, Tomoki; Kimura, Makoto; Asakawa, Susumu

    2015-01-01

    Methanogenic archaea are strict anaerobes and demand highly reduced conditions to produce methane in paddy field soil. However, methanogenic archaea survive well under upland and aerated conditions in paddy fields and exhibit stable community. In the present study, methanogenic archaeal community was investigated in fields where paddy rice (Oryza sativa L.) under flooded conditions was rotated with soybean (Glycine max [L.] Merr.) under upland conditions at different rotation histories, by polymerase chain reaction (PCR)-denaturing gradient gel electrophoresis (DGGE) and real-time quantitative PCR methods targeting 16S rRNA and mcrA genes, respectively. Soil samples collected from the fields before flooding or seeding, during crop cultivation and after harvest of crops were analyzed. The abundance of the methanogenic archaeal populations decreased to about one-tenth in the rotational plots than in the consecutive paddy (control) plots. The composition of the methanogenic archaeal community also changed. Most members of the methanogenic archaea consisting of the orders Methanosarcinales, Methanocellales, Methanomicrobiales, and Methanobacteriales existed autochthonously in both the control and rotational plots, while some were strongly affected in the rotational plots, with fatal effect to some members belonging to the Methanosarcinales. This study revealed that the upland conversion for one or longer than 1 year in the rotational system affected the methanogenic archaeal community structure and was fatal to some members of methanogenic archaea in paddy field soil. PMID:25113614

  5. Sleeping distance in wild wolf packs

    USGS Publications Warehouse

    Knick, S.T.; Mech, L.D.

    1980-01-01

    Sleeping distances were observed among members of 13 wild wolf (Canis lupus) packs and 11 pairs in northeastern Minnesota to determine if the distances correlated with pack size and composition. The study utilized aerial radio-tracking and observation during winter. Pack size and number of adults per pack were inversely related to pack average sleeping distance and variability. No correlation between sleeping distance and microclimate was observed. Possible relationships between social bonding and our results are discussed.

  6. Two-phase bioreactor system for cell-laden hydrogel assembly.

    PubMed

    Gulfam, Muhammad; Lee, Jong Min; Chung, Bong Geun

    2011-01-01

    Bottom-up approach is a potentially useful tool for hydrogel assembly of cell-laden individual building blocks. In this article, we assembled individual building blocks of photocrosslinkable microgels in a rapid and controlled manner. Individual building blocks of poly(ethylene glycol) (PEG) microgels with square and hexagonal shapes were fabricated by using a photolithography technique. Individual building blocks of PEG microgels were assembled on a hydrophobic mineral oil phase in a bioreactor with a magnetic stirrer. The hydrophobic mineral oil minimized the surface free energy to assemble hydrophilic PEG microgels on a two-phase oil-aqueous solution interface. We used the hydrophobic effect as a driving force for the hydrogel assembly. Various types of the hydrogel assembly were generated by controlling the stirring rate. As stirring speed increased, the percentage of linear, branched, and closely packed hydrogel assembly was increased. However, the percentage of random assembly was reduced by increasing stirring rate. The stirring time also played an important role in controlling the types of hydrogel assembly. The percentage of linear, branched, and closely packed hydrogel assembly was improved by increasing stirring time. Therefore, we performed directed cell-laden hydrogel assembly using a two-phase bioreactor system and optimized the stirring rate and time to regulate the desired types of hydrogel assembly. Furthermore, we analyzed cell viability of hydrogel linear assembly with square shapes, showing highly viable even after secondary photocrosslinking reaction. This bioreactor system-based hydrogel assembly could be a potentially powerful approach for creating tissue microarchitectures in a three-dimensional manner.

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

    PubMed

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

    2010-01-01

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

  8. Quantitative Immunologic Analysis of the Methanogenic Flora of Digestors Reveals a Considerable Diversity

    PubMed Central

    Macario, Alberto J. L.; de Macario, Everly Conway

    1988-01-01

    To determine which methanogens occur in digestors, we performed a quantitative immunologic analysis of a variety of samples. A comprehensive panel of calibrated polyclonal antibody probes of predefined specificity spectra was used. This allowed precise identification of bacteria by antigenic fingerprinting. A considerable diversity of methanogens was uncovered, much larger than previously reported, encompassing at least 14 strains of 11 species. Strategies were developed to measure the load of any given methanogen in a sample and to compare samples quantitatively. Two methanogens were found to predominate which were antigenically closely related with either Methanobacterium formicicum MF or Methanobrevibacter arboriphilus AZ. Fundamental data, probes, and methods are now available to monitor methanogenic subpopulations during digestor operation and thus learn about their respective roles and predictive significance. Images PMID:16347541

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

    PubMed Central

    Narihiro, Takashi; Sekiguchi, Yuji

    2011-01-01

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

  10. Methane formation and methane oxidation by methanogenic bacteria.

    PubMed Central

    Zehnder, A J; Brock, T D

    1979-01-01

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

  11. Presence of an Unusual Methanogenic Bacterium in Coal Gasification Waste

    PubMed Central

    Tomei, Francisco A.; Rouse, Dwight; Maki, James S.; Mitchell, Ralph

    1988-01-01

    Methanogenic bacteria growing on a pilot-scale, anaerobic filter processing coal gasification waste were enriched in a mineral salts medium containing hydrogen and acetate as potential energy sources. Transfer of the enrichments to methanol medium resulted in the initial growth of a strain of Methanosarcina barkeri, but eventually small cocci became dominant. The cocci growing on methanol produced methane and exhibited the typical fluorescence of methanogenic bacteria. They grew in the presence of the cell wall synthesis-inhibiting antibiotics d-cycloserine, fosfomycin, penicillin G, and vancomycin as well as in the presence of kanamycin, an inhibitor of protein synthesis in eubacteria. The optimal growth temperature was 37°C, and the doubling time was 7.5 h. The strain lysed after reaching stationary phase. The bacterium grew poorly with hydrogen as the energy source and failed to grow on acetate. Morphologically, the coccus shared similarities with Methanosarcina sp. Cells were 1 μm wide, exhibited the typical thick cell wall and cross-wall formation, and formed tetrads. Packets and cysts were not formed. Images PMID:16347791

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

    PubMed Central

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

    2016-01-01

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

  13. Interactions of methanogens and denitrifiers in degradation of phenols

    SciTech Connect

    Fang, H.H.P.; Zhou, G.M.

    1999-01-01

    Experiments were conducted at 37 C in an upflow anaerobic sludge blanket reactor treating wastewater containing phenol, m-cresol, and nitrate at various concentrations. Results show that anaerobic sludge was able to conduct denitrification without much acclimation. Denitrifiers outcompeted methanogens for substrates for carbon and electron supplies. They were able to use phenol and m-cresol as substrate without a carbohydrate cosubstrate. Denitrifying 1 g of NO{sub 3}{sup {minus}}-N ratios greater than 3.34. At the ratio of 5.23, over 98% of phenol but only 60% of m-cresol were degraded jointly by denitrifiers and methanogens with 1 day of hydraulic retention. At ratios less than 3.34, methanogenesis ceased to take place and denitrification became incomplete because of insufficient supply of substrate. Batch tests further confirmed that degradation of m-cresol was enhanced not only by the presence of nitrate, but also by the presence of either sucrose or phenol as cosubstrate.

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

    PubMed

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

    2016-01-01

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

  15. Entanglement Theories: Packing vs. Percolation

    NASA Astrophysics Data System (ADS)

    Wool, Richard

    2007-03-01

    There are two emergent theories of polymer entanglements, the Packing Model (Fetters, Lohse, Graessley, Milner, Whitten, ˜'98) and the Percolation Model (Wool ˜'93). The Packing model suggests that the entanglement molecular weight Me is determined by Me = K p^3, where the packing length parameter p = V/R^2 in which V is the volume of the chain (V=M/ρNa), R is the end-to end vector of the chain, and K 357 ρNa, is an empirical constant. The Percolation model states that an entanglement network develops when the number of chains per unit area σ, intersecting any load bearing plane, is equal to 3 times the number of chain segments (1/a cross-section), such that when 3aσ =1 at the percolation threshold, Me 31 MjC∞, in which Mj is the step molecular weight and C∞ is the characteristic ratio. There are no fitting parameters in the Percolation model. The Packing model predicts that Me decreases rapidly with chain stiffness, as Me˜1/C∞^3, while the Percolation model predicts that Me increases with C∞, as Me˜C∞. The Percolation model was found to be the correct model based on computer simulations (M. Bulacu et al) and a re-analysis of the Packing model experimental data. The Packing model can be derived from the Percolation model, but not visa versa, and reveals a surprising accidental relation between C∞ and Mj in the front factor K. This result significantly impacts the interpretation of the dynamics of rheology and fracture of entangled polymers.

  16. Open source software to control Bioflo bioreactors.

    PubMed

    Burdge, David A; Libourel, Igor G L

    2014-01-01

    Bioreactors are designed to support highly controlled environments for growth of tissues, cell cultures or microbial cultures. A variety of bioreactors are commercially available, often including sophisticated software to enhance the functionality of the bioreactor. However, experiments that the bioreactor hardware can support, but that were not envisioned during the software design cannot be performed without developing custom software. In addition, support for third party or custom designed auxiliary hardware is often sparse or absent. This work presents flexible open source freeware for the control of bioreactors of the Bioflo product family. The functionality of the software includes setpoint control, data logging, and protocol execution. Auxiliary hardware can be easily integrated and controlled through an integrated plugin interface without altering existing software. Simple experimental protocols can be entered as a CSV scripting file, and a Python-based protocol execution model is included for more demanding conditional experimental control. The software was designed to be a more flexible and free open source alternative to the commercially available solution. The source code and various auxiliary hardware plugins are publicly available for download from https://github.com/LibourelLab/BiofloSoftware. In addition to the source code, the software was compiled and packaged as a self-installing file for 32 and 64 bit windows operating systems. The compiled software will be able to control a Bioflo system, and will not require the installation of LabVIEW. PMID:24667828

  17. Open source software to control Bioflo bioreactors.

    PubMed

    Burdge, David A; Libourel, Igor G L

    2014-01-01

    Bioreactors are designed to support highly controlled environments for growth of tissues, cell cultures or microbial cultures. A variety of bioreactors are commercially available, often including sophisticated software to enhance the functionality of the bioreactor. However, experiments that the bioreactor hardware can support, but that were not envisioned during the software design cannot be performed without developing custom software. In addition, support for third party or custom designed auxiliary hardware is often sparse or absent. This work presents flexible open source freeware for the control of bioreactors of the Bioflo product family. The functionality of the software includes setpoint control, data logging, and protocol execution. Auxiliary hardware can be easily integrated and controlled through an integrated plugin interface without altering existing software. Simple experimental protocols can be entered as a CSV scripting file, and a Python-based protocol execution model is included for more demanding conditional experimental control. The software was designed to be a more flexible and free open source alternative to the commercially available solution. The source code and various auxiliary hardware plugins are publicly available for download from https://github.com/LibourelLab/BiofloSoftware. In addition to the source code, the software was compiled and packaged as a self-installing file for 32 and 64 bit windows operating systems. The compiled software will be able to control a Bioflo system, and will not require the installation of LabVIEW.

  18. Open Source Software to Control Bioflo Bioreactors

    PubMed Central

    Burdge, David A.; Libourel, Igor G. L.

    2014-01-01

    Bioreactors are designed to support highly controlled environments for growth of tissues, cell cultures or microbial cultures. A variety of bioreactors are commercially available, often including sophisticated software to enhance the functionality of the bioreactor. However, experiments that the bioreactor hardware can support, but that were not envisioned during the software design cannot be performed without developing custom software. In addition, support for third party or custom designed auxiliary hardware is often sparse or absent. This work presents flexible open source freeware for the control of bioreactors of the Bioflo product family. The functionality of the software includes setpoint control, data logging, and protocol execution. Auxiliary hardware can be easily integrated and controlled through an integrated plugin interface without altering existing software. Simple experimental protocols can be entered as a CSV scripting file, and a Python-based protocol execution model is included for more demanding conditional experimental control. The software was designed to be a more flexible and free open source alternative to the commercially available solution. The source code and various auxiliary hardware plugins are publicly available for download from https://github.com/LibourelLab/BiofloSoftware. In addition to the source code, the software was compiled and packaged as a self-installing file for 32 and 64 bit windows operating systems. The compiled software will be able to control a Bioflo system, and will not require the installation of LabVIEW. PMID:24667828

  19. Small-scale, hydrogen-oxidizing-denitrifying bioreactor for treatment of nitrate-contaminated drinking water

    USGS Publications Warehouse

    Smith, R.L.; Buckwalter, S.P.; Repert, D.A.; Miller, D.N.

    2005-01-01

    Nitrate removal by hydrogen-coupled denitrification was examined using flow-through, packed-bed bioreactors to develop a small-scale, cost effective system for treating nitrate-contaminated drinking-water supplies. Nitrate removal was accomplished using a Rhodocyclus sp., strain HOD 5, isolated from a sole-source drinking-water aquifer. The autotrophic capacity of the purple non-sulfur photosynthetic bacterium made it particularly adept for this purpose. Initial tests used a commercial bioreactor filled with glass beads and countercurrent, non-sterile flow of an autotrophic, air-saturated, growth medium and hydrogen gas. Complete removal of 2 mM nitrate was achieved for more than 300 days of operation at a 2-h retention time. A low-cost hydrogen generator/bioreactor system was then constructed from readily available materials as a water treatment approach using the Rhodocyclus strain. After initial tests with the growth medium, the constructed system was tested using nitrate-amended drinking water obtained from fractured granite and sandstone aquifers, with moderate and low TDS loads, respectively. Incomplete nitrate removal was evident in both water types, with high-nitrite concentrations in the bioreactor output, due to a pH increase, which inhibited nitrite reduction. This was rectified by including carbon dioxide in the hydrogen stream. Additionally, complete nitrate removal was accomplished with wastewater-impacted surface water, with a concurrent decrease in dissolved organic carbon. The results of this study using three chemically distinct water supplies demonstrate that hydrogen-coupled denitrification can serve as the basis for small-scale remediation and that pilot-scale testing might be the next logical step.

  20. Small-scale, hydrogen-oxidizing-denitrifying bioreactor for treatment of nitrate-contaminated drinking water.

    PubMed

    Smith, Richard L; Buckwalter, Seanne P; Repert, Deborah A; Miller, Daniel N

    2005-05-01

    Nitrate removal by hydrogen-coupled denitrification was examined using flow-through, packed-bed bioreactors to develop a small-scale, cost effective system for treating nitrate-contaminated drinking-water supplies. Nitrate removal was accomplished using a Rhodocyclus sp., strain HOD 5, isolated from a sole-source drinking-water aquifer. The autotrophic capacity of the purple non-sulfur photosynthetic bacterium made it particularly adept for this purpose. Initial tests used a commercial bioreactor filled with glass beads and countercurrent, non-sterile flow of an autotrophic, air-saturated, growth medium and hydrogen gas. Complete removal of 2 mM nitrate was achieved for more than 300 days of operation at a 2-h retention time. A low-cost hydrogen generator/bioreactor system was then constructed from readily available materials as a water treatment approach using the Rhodocyclus strain. After initial tests with the growth medium, the constructed system was tested using nitrate-amended drinking water obtained from fractured granite and sandstone aquifers, with moderate and low TDS loads, respectively. Incomplete nitrate removal was evident in both water types, with high-nitrite concentrations in the bioreactor output, due to a pH increase, which inhibited nitrite reduction. This was rectified by including carbon dioxide in the hydrogen stream. Additionally, complete nitrate removal was accomplished with wastewater-impacted surface water, with a concurrent decrease in dissolved organic carbon. The results of this study using three chemically distinct water supplies demonstrate that hydrogen-coupled denitrification can serve as the basis for small-scale remediation and that pilot-scale testing might be the next logical step.

  1. Nitrogen management in bioreactor landfills

    SciTech Connect

    Price, G. Alexander; Barlaz, Morton A.; Hater, Gary R

    2003-07-01

    One scenario for long-term nitrogen management in landfills is ex situ nitrification followed by denitrification in the landfill. The objective of this research was to measure the denitrification potential of actively decomposing and well decomposed refuse. A series of 10-l reactors that were actively producing methane were fed 400 mg NO{sub 3}-N /l every 48 h for periods of 19-59 days. Up to 29 nitrate additions were either completely or largely depleted within 48 h of addition and the denitrification reactions did not adversely affect the leachate pH. Nitrate did inhibit methane production, but the reactors recovered their methane-producing activity with the termination of nitrate addition. In well decomposed refuse, the nitrate consumption rate was reduced but was easily stimulated by the addition of either acetate or an overlayer of fresh refuse. Addition of acetate at five times the amount required to reduce nitrate did not lead to the production of NH{sub 4}{sup +} by dissimilatory nitrate reduction. The most probable number of denitrifying bacteria decreased by about five orders of magnitude during refuse decomposition in a reactor that did not receive nitrate. However, rapid denitrification commenced immediately with nitrate addition. This study shows that the use of a landfill as a bioreactor for the conversion of nitrate to a harmless byproduct, nitrogen gas, is technically viable.

  2. Bioreactor control improves bioprocess performance.

    PubMed

    Simutis, Rimvydas; Lübbert, Andreas

    2015-08-01

    The performance of bioreactors is not only determined by productivity but also by process quality, which is mainly determined by variances in the process variables. As fluctuations in these quantities directly affect the variability in the product properties, combatting distortions is the main task of practical quality assurance. The straightforward way of reducing this variability is keeping the product formation process tightly under control. Purpose of this keynote is to show that there is enough evidence in literature showing that the performance of the fermentation processes can significantly be improved by feedback control. Most of the currently used open loop control procedures can be replaced by relatively simple feedback techniques. It is shown by practical examples that such a retrofitting does not require significant changes in the well-established equipment. Feedback techniques are best in assuring high reproducibility of the industrial cultivation processes and thus in assuring the quality of their products. Many developments in supervising and controlling industrial fermentations can directly be taken over in manufacturing processes. Even simple feedback controllers can efficiently improve the product quality. It's the time now that manufacturers follow the developments in most other industries and improve process quality by automatic feedback control. PMID:26228573

  3. Bioreactor control improves bioprocess performance.

    PubMed

    Simutis, Rimvydas; Lübbert, Andreas

    2015-08-01

    The performance of bioreactors is not only determined by productivity but also by process quality, which is mainly determined by variances in the process variables. As fluctuations in these quantities directly affect the variability in the product properties, combatting distortions is the main task of practical quality assurance. The straightforward way of reducing this variability is keeping the product formation process tightly under control. Purpose of this keynote is to show that there is enough evidence in literature showing that the performance of the fermentation processes can significantly be improved by feedback control. Most of the currently used open loop control procedures can be replaced by relatively simple feedback techniques. It is shown by practical examples that such a retrofitting does not require significant changes in the well-established equipment. Feedback techniques are best in assuring high reproducibility of the industrial cultivation processes and thus in assuring the quality of their products. Many developments in supervising and controlling industrial fermentations can directly be taken over in manufacturing processes. Even simple feedback controllers can efficiently improve the product quality. It's the time now that manufacturers follow the developments in most other industries and improve process quality by automatic feedback control.

  4. [Electroflotation packing of mycelial wastes].

    PubMed

    Nikolaev, V B; Karpukhin, V F; Zav'ialova, E V; Faingol'd, Z L

    1989-04-01

    Data on testing a pilot plant for electroflotative packing of mycelial wastes are presented. Both the mycelial wastes and their mixtures were used. Concentration of dry substances raged from 3.5 to 131 g/l. The process was performed with varying flow density and consumption rate of the liquid supplied to the plant. Insoluble magnetite and ruthenium oxide anodes were used. The moister content in the packed phase was 81-97.5 per cent. The study provided specification of the process technological parameters and investigation of the flotator hydroulic characteristics.

  5. Cylinder valve packing nut studies

    SciTech Connect

    Blue, S.C.

    1991-12-31

    The design, manufacture, and use of cylinder valve packing nuts have been studied to improve their resistance to failure from stress corrosion cracking. Stress frozen photoelastic models have been analyzed to measure the stress concentrations at observed points of failure. The load effects induced by assembly torque and thermal expansion of stem packing were observed by strain gaging nuts. The effects of finishing operations and heat treatment were studied by the strain gage hole boring and X-ray methods. Modifications of manufacturing and operation practices are reducing the frequency of stress corrosion failures.

  6. Two-stage anaerobic membrane bioreactor for the treatment of sugarcane vinasse: assessment on biological activity and filtration performance.

    PubMed

    Mota, Vera Tainá; Santos, Fábio S; Amaral, Míriam C S

    2013-10-01

    A two-stage submerged anaerobic membrane bioreactor (2-SAnMBR) was designed for the treatment of sugarcane vinasse. For start-up, the flow rate was reduced whenever VFA levels reached critical levels in the methanogenic reactor. After acclimation, the system was operated under a continuous flow. Separation of the stages was observed during the entire period of operation. VFA, COD and DOC levels of raw effluent, acidified effluent and permeate averaged 2141, 3525 and 61 mg VFA L(-1) (as acetic acid), 15727, 11512 and 488 mg COD L(-1), and, 3544, 3533 and 178 mg DOC L(-1), respectively. Overall COD and DOC removal efficiencies of 96.9±0.7% and 95.0±1.1%, respectively, were reached. Methane content of the biogas from the acidogenic and methanogenic reactors ranged 0.1-4.6% and 60.1-70.1%, respectively. Removable fouling strongly affected filtration performance and cake layer formation accounted for most of filtration resistance. Membrane resistance was related to presence of protein-like substances and carbohydrates. PMID:23958682

  7. Hydrolysis, acidification and methanogenesis during low-temperature anaerobic digestion of dilute dairy wastewater in an inverted fluidised bioreactor.

    PubMed

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

    2014-10-01

    The application of low-temperature (10 °C) anaerobic digestion (LtAD) for the treatment of complex dairy-based wastewater in an inverted fluidised bed (IFB) reactor was investigated. Inadequate mixing intensity provoked poor hydrolysis of the substrate (mostly protein), which resulted in low chemical oxygen demand (COD) removal efficiency throughout the trial, averaging ~69 % at the best operational period. Overgrowth of the attached biomass to the support particles (Extendospheres) induced bed stratification by provoking agglutination of the particles and supporting their washout by sedimentation, which contributed to unstable bioprocess performance at the organic loading rates (OLRs) between 0.5 and 5 kg COD m(-3) day(-1). An applied OLR above 2 kg COD m(-3) day(-1) additionally promoted acidification and strongly influenced the microbial composition and dynamics. Hydrogenotrophic methanogens appeared to be the mostly affected group by the Extendospheres particle washout as a decrease in their abundance was observed by quantitative PCR analysis towards the end of the trial, although the specific methanogenic activity and maximum substrate utilisation rate on H2/CO2 indicated high metabolic activity and preference towards hydrogenotrophic methanogenesis of the reactor biomass at this stage. The bacterial community in the bioreactor monitored via denaturing gradient gel electrophoresis (DGGE) also suggested an influence of OLR stress on bacterial community structure and population dynamics. The data presented in this work can provide useful information in future optimisation of fluidised reactors intended for digestion of complex industrial wastewaters during LtAD. PMID:24946864

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

    PubMed Central

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

    2016-01-01

    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. PMID:26830464

  9. Hydrolysis, acidification and methanogenesis during low-temperature anaerobic digestion of dilute dairy wastewater in an inverted fluidised bioreactor.

    PubMed

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

    2014-10-01

    The application of low-temperature (10 °C) anaerobic digestion (LtAD) for the treatment of complex dairy-based wastewater in an inverted fluidised bed (IFB) reactor was investigated. Inadequate mixing intensity provoked poor hydrolysis of the substrate (mostly protein), which resulted in low chemical oxygen demand (COD) removal efficiency throughout the trial, averaging ~69 % at the best operational period. Overgrowth of the attached biomass to the support particles (Extendospheres) induced bed stratification by provoking agglutination of the particles and supporting their washout by sedimentation, which contributed to unstable bioprocess performance at the organic loading rates (OLRs) between 0.5 and 5 kg COD m(-3) day(-1). An applied OLR above 2 kg COD m(-3) day(-1) additionally promoted acidification and strongly influenced the microbial composition and dynamics. Hydrogenotrophic methanogens appeared to be the mostly affected group by the Extendospheres particle washout as a decrease in their abundance was observed by quantitative PCR analysis towards the end of the trial, although the specific methanogenic activity and maximum substrate utilisation rate on H2/CO2 indicated high metabolic activity and preference towards hydrogenotrophic methanogenesis of the reactor biomass at this stage. The bacterial community in the bioreactor monitored via denaturing gradient gel electrophoresis (DGGE) also suggested an influence of OLR stress on bacterial community structure and population dynamics. The data presented in this work can provide useful information in future optimisation of fluidised reactors intended for digestion of complex industrial wastewaters during LtAD.

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

    NASA Astrophysics Data System (ADS)

    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.

  11. Microbial dynamics in upflow anaerobic sludge blanket (UASB) bioreactor granules in response to short-term changes in substrate feed

    SciTech Connect

    Kovacik, William P.; Scholten, Johannes C.; Culley, David E.; Hickey, Robert; Zhang, Weiwen; Brockman, Fred J.

    2010-08-01

    The complexity and diversity of the microbial communities in biogranules from an upflow anaerobic sludge blanket (UASB) bioreactor were determined in response to short-term changes in substrate feeds. The reactor was fed simulated brewery wastewater (SBWW) (70% ethanol, 15% acetate, 15% propionate) for 1.5 months (phase 1), acetate / sulfate for 2 months (phase 2), acetate-alone for 3 months (phase 3), and then a return to SBWW for 2 months (phase 4). Performance of the reactor remained relatively stable throughout the experiment as shown by COD removal and gas production. 16S rDNA, methanogen-associated mcrA and sulfate reducer-associated dsrAB genes were PCR amplified, then cloned and sequenced. Sequence analysis of 16S clone libraries showed a relatively simple community composed mainly of the methanogenic Archaea (Methanobacterium and Methanosaeta), members of the Green Non-Sulfur (Chloroflexi) group of Bacteria, followed by fewer numbers of Syntrophobacter, Spirochaeta, Acidobacteria and Cytophaga-related Bacterial sequences. Methanogen-related mcrA clone libraries were dominated throughout by Methanobacter and Methanospirillum related sequences. Although not numerous enough to be detected in our 16S rDNA libraries, sulfate reducers were detected in dsrAB clone libraries, with sequences related to Desulfovibrio and Desulfomonile. Community diversity levels (Shannon-Weiner index) generally decreased for all libraries in response to a change from SBWW to acetate-alone feed. But there was a large transitory increase noted in 16S diversity at the two-month sampling on acetate-alone, entirely related to an increase in Bacterial diversity. Upon return to SBWW conditions in phase 4, all diversity measures returned to near phase 1 levels.

  12. Design concepts for bioreactors in space

    NASA Technical Reports Server (NTRS)

    Seshan, P. K.; Peterson, G. R.; Beard, B.; Dunlop, E. H.

    1986-01-01

    Microbial food sources are becoming viable and more efficient alternatives to conventional food sources especially in the context of Closed Ecological Life Support Systems (CELSS) in space habitats. Since bioreactor designs for terrestrial operation will not readily apply to conditions of microgravity, there is an urgent need to learn about the differences. These differences cannot be easily estimated due to the complex nature of the mass transport and mixing mechanisms in fermenters. Therefore, a systematic and expeditious experimental program must be undertaken to obtain the engineering data necessary to lay down the foundations of designing bioreactors for microgravity. Two bioreactor design concepts presented represent two dissimilar approaches to grappling with the absence of gravity in space habitats and deserve to be tested for adoption as important components of the life support function aboard spacecrafts, space stations and other extra-terrestrial habitats.

  13. A nanoliter microfluidic serial dilution bioreactor.

    PubMed

    Gu, Guo-Yue; Lee, Yi-Wei; Chiang, Chih-Chung; Yang, Ya-Tang

    2015-07-01

    Bacterial culture is a basic technique in both fundamental and applied microbiology. The excessive reagent consumption and laborious maintenance of bulk bioreactors for microbial culture have prompted the development of miniaturized on-chip bioreactors. With the minimal choice of two compartments (N = 2) and discrete time, periodic dilution steps, we realize a microfluidic bioreactor that mimics macroscopic serial dilution transfer culture. This device supports automated, long-term microbial cultures with a nanoliter-scale working volume and real-time monitoring of microbial populations at single-cell resolution. Because of the high surface-to-volume ratio, the device also operates as an effective biofilm-flow reactor to support cogrowth of planktonic and biofilm populations. We expect that such devices will open opportunities in many fields of microbiology.

  14. Ecology and Energy Action Pack.

    ERIC Educational Resources Information Center

    McDonald's Corp., Oak Brook, IL.

    One of five McDonald's Action Packs, these elementary school-level instructional materials are for use as an introduction to existing units of study, supplements to a textbook, or a source of special projects for environmental education. Contents include these six units: Make Your Own Ecology Mini-spinner, Let's Look at a Food Chain, Drip the…

  15. Pack cementation coatings for alloys

    SciTech Connect

    He, Yi-Rong; Zheng, Minhui; Rapp, R.A.

    1996-08-01

    The halide-activated pack cementation process was modified to produce a Ge-doped silicide diffusion coating on a Cr-Cr{sub 2}Nb alloy in a single processing step. The morphology and composition of the coating depended both on the composition of the pack and on the composition and microstructure of the substrate. Higher Ge content in the pack suppressed the formation of CrSi{sub 2} and reduced the growth kinetics of the coating. Ge was not homogeneously distributed in the coatings. In cyclic and isothermal oxidation in air at 700 and 1050{degrees}C, the Ge-doped silicide coating protected the Cr-Nb alloys from significant oxidation by the formation of a Ge-doped silica film. The codeposition and diffusion of aluminum and chromium into low alloy steel have been achieved using elemental Al and Cr powders and a two-step pack cementation process. Sequential process treatments at 925{degrees}C and 1150{degrees}C yield dense and uniform ferrite coatings, whose compositions are close to either Fe{sub 3}Al or else FeAl plus a lower Cr content, when processed under different conditions. The higher content of Al in the coatings was predicted by thermodynamic calculations of equilibrium in the gas phase. The effect of the particle size of the metal powders on the surface composition of the coating has been studied for various combinations of Al and Cr powders.

  16. Wire and Packing Tape Sandwiches

    ERIC Educational Resources Information Center

    Rabinowitz, Sandy

    2009-01-01

    In this article, the author describes how students can combine craft wire with clear packing tape to create a two-dimensional design that can be bent and twisted to create a three-dimensional form. Students sandwich wire designs between two layers of tape. (Contains 1 online resource.)

  17. Bioreactor and methods for producing synchronous cells

    NASA Technical Reports Server (NTRS)

    Helmstetter, Charles E. (Inventor); Thornton, Maureen (Inventor); Gonda, Steve (Inventor)

    2005-01-01

    Apparatus and methods are directed to a perfusion culture system in which a rotating bioreactor is used to grow cells in a liquid culture medium, while these cells are attached to an adhesive-treated porous surface. As a result of this arrangement and its rotation, the attached cells divide, with one cell remaining attached to the substrate, while the other cell, a newborn cell is released. These newborn cells are of approximately the same age, that are collected upon leaving the bioreactor. The populations of newborn cells collected are of synchronous and are minimally, if at all, disturbed metabolically.

  18. Development of a Space Bioreactor using Microtechnology

    NASA Technical Reports Server (NTRS)

    Arquint, Philippe; Boillat, Marc A.; deRooij, Nico F.; Jeanneret, Sylvain; vanderSchoot, Bart H.; Bechler, Birgitt; Cogoli, Augusto; Walther, Isabelle; Gass, Volker; Ivorra, Marie-Therese

    1995-01-01

    A miniature bio-reactor for the cultivation of cells aboard Spacelab is presented. Yeast cells are grown in a 3 milliliter reactor chamber. A supply of fresh nutrient medium is provided by a piezo-electric silicon micro-pump. In the reactor, pH, temperature, and redox potential are monitored and the pH is regulated at a constant value. The complete instrument is fitted in a standard experiment container of 63 x 63 x 85 mm. The bioreactor was used on the IML-2 mission in July 1994 and is being refurbished for a reflight in the spring of 1996.

  19. Transport and kinetics in sandwiched membrane bioreactors.

    PubMed

    Jeong, Y S; Vieth, W R; Matsuura, T

    1991-01-01

    A bioreactor in which living yeast cells are sandwiched between an ultrafiltration membrane and a reverse osmosis membrane was constructed, and experiments were performed for the conversion of substrate glucose to product ethanol. A set of equations that include both transport through a series of barrier layers and bioreaction rate were developed to predict the performance of the sandwich bioreactor. The above equations were solved by using numerical values for the transport parameter and the bioreaction rate constant, and the results are compared with the experimental data.

  20. Molecular phylogeny of methanogens associated with flagellated protists in the gut and with the gut epithelium of termites.

    PubMed

    Tokura; Ohkuma; Kudo

    2000-09-01

    The molecular phylogeny of methanogenic archaea associated with the flagellated protist species Dinenympha and Microjoenia in the gut of termites, Reticulitermes speratus and Hodotermopsis sjoestedti, and those attached to the gut epithelium was examined based on PCR-amplified small-subunit ribosomal RNA genes. The sequences identified were classified into six groups within the genus Methanobrevibacter, including groups of yet uncharacterized novel species. Closely related methanogens were shared between Microjoenia and some Dinenympha cells in each termite. The methanogens harbored by the flagellates were phylogenetically different from the methanogens associated with the gut epithelium, suggesting that distinct methanogen species showed distinct spatial distributions in the termite gut.

  1. Cultivation of methanogens from shallow marine sediments at Hydrate Ridge, Oregon

    PubMed Central

    Kendall, Melissa M.; Boone, David R.

    2006-01-01

    Little is known about the methanogenic degradation of acetate, the fate of molecular hydrogen and formate or the ability of methanogens to grow and produce methane in cold, anoxic marine sediments. The microbes that produce methane were examined in permanently cold, anoxic marine sediments at Hydrate Ridge (44°35' N, 125°10' W, depth 800 m). Sediment samples (15 to 35 cm deep) were collected from areas of active methane ebullition or areas where methane hydrates occurred. The samples were diluted into enrichment medium with formate, acetate or trimethylamine as catabolic substrate. After 2 years of incubation at 4 °C to 15 °C, enrichment cultures produced methane. PCR amplification and sequencing of the rRNA genes from the highest dilutions with growth suggested that each enrichment culture contained a single strain of methanogen. The level of sequence similarity (91 to 98%) to previously characterized prokaryotes suggested that these methanogens belonged to novel genera or species within the orders Methanomicrobiales and Methanosarcinales. Analysis of the 16S rRNA gene libraries from DNA extracted directly from the sediment samples revealed phylotypes that were either distantly related to cultivated methanogens or possible anaerobic methane oxidizers related to the ANME-1 and ANME-2 groups of the Archaea. However, no methanogenic sequences were detected, suggesting that methanogens represented only a small proportion of the archaeal. PMID:16877319

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

    SciTech Connect

    Kemp, C.W.

    1985-01-01

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

  3. A multicommutated tester of bioreactors for flow analysis.

    PubMed

    Pokrzywnicka, Marta; Kamiński, Jacek; Michalec, Michał; Koncki, Robert; Tymecki, Łukasz

    2016-11-01

    Enzymes are often used in the modern analytical procedures allowing selective recognition and conversion of target analytes into easily detected products. In flow analysis systems, enzymes are predominantly applied in the immobilized forms as flow-through bioreactors. In this research the multicommutated flow analysis (MCFA) system for evaluation and comparison of analytical parameters of bioreactors has been developed. The MCFA manifold allows simultaneous testing up to four bioreactors, but if necessary their number can be easily increased. The system allows comparison of several parameters of tested bioreactors including activity, repeatability, reproducibility, operational and storage stability. The performance of developed bioreactor tester is presented using urea-urease model system based on plastic open-tubular bioreactor with covalently immobilized enzyme. Product of enzymatic reaction is detected using two different chemical methods and by dedicated optoelectronic ammonium detectors. Moreover, the utility of developed MCFA manifold for evaluation of other enzyme bioreactors is demonstrated. PMID:27591609

  4. Packing tube assembly for pumping wells

    SciTech Connect

    Towner, G.F.; Carter, C.A.

    1987-09-22

    A packing tube assembly for replacing a conventional stuffing box is described. The packing tube assembly comprising: a packing tube; a rod adaptor adapted to be coupled between the polished rod and the sucker rod string and adapted to extend throughout the packing tube when positioned therein; compressible packing means on the rod adaptor adapted to provide a seal between the rod adaptor and the packing tube when the rod adaptor is in position within the packing tube; stabilizing means on the rod adaptor adapted to engage the packing tube to stabilize the rod adaptor within the packing tube during operation; and a mounting bushing connected to the top of the packing tube and adapted to be threaded into the pumping tee to secure the packing tube position within the production tubing string. A method of converting a conventional stuffing box-equipped pumping well to a packing tube-equipped well is described. It consists of: disconnecting the polished rod of the pumping well from the sucker rod string while suspending the sucker rod string within the well by the use of slips; unthreading the conventional stuffing box from the pumping tee of the well and removing the stuffing box; attaching the polished rod to the upper end of a rod adaptor of a packing tube assembly, inserting the rod adaptor with the polished rod attached into the packing tube of the packing tube assembly; aligning the packing tube assembly with the sucker rod string; connecting the lower end of the rod adaptor to the sucker rod string; removing the slips and lowering the packer tube assembly through the pumping tee; and connecting the upper end of the packing tube to the pumping tee.

  5. Methanogenic burst in the end-Permian carbon cycle

    PubMed Central

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

    2014-01-01

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

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

    DOE PAGES

    Rother, Michael; Krzycki, Joseph A.

    2010-01-01

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

  7. Effect of acclimation on methanogenic degradation of chlorophenols

    SciTech Connect

    Wang, Y.T.; Muthukrishnan, S.

    1996-11-01

    Chlorophenols are highly toxic and persistent in the environment. Several millions of pounds of chlorinated phenols and chlorophenol based compounds are manufactured and used every year. Pentachlorophenol (PCP) and tetrachlorophenols (TCP) are widely used in the paper pulp industry and also as wood preservatives. Chlorophenols are also formed during the disinfection of wastewater containing phenols and in chlorine bleaching processes of cellulose. Anaerobic biodegradation of chlorophenols by anaerobic microbial consortia has been extensively studied by many researchers. Anaerobic biodegradation of chlorophenols occurs through a series of reductive dechlorination, in which the chlorine is replaced by hydrogen at each step. This reductive dehalogenation is of environmental importance because the less chlorinated metabolic products are generally less toxic and more easily degraded by aerobic bacteria. The main objective of this study is to examine the degradation of chlorophenols in both unacclimated and acclimated methanogenic cultures.

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

    PubMed Central

    Rother, Michael; Krzycki, Joseph A.

    2010-01-01

    Methanogenic archaea are a group of strictly anaerobic microorganisms characterized by their strict dependence on the process of methanogenesis for energy conservation. Among the archaea, they are also the only known group synthesizing proteins containing selenocysteine or pyrrolysine. All but one of the known archaeal pyrrolysine-containing and all but two of the confirmed archaeal selenocysteine-containing protein are involved in methanogenesis. Synthesis of these proteins proceeds through suppression of translational stop codons but otherwise the two systems are fundamentally different. This paper highlights these differences and summarizes the recent developments in selenocysteine- and pyrrolysine-related research on archaea and aims to put this knowledge into the context of their unique energy metabolism. PMID:20847933

  9. Methane production from coal by a single methanogen

    NASA Astrophysics Data System (ADS)

    Mayumi, Daisuke; Mochimaru, Hanako; Tamaki, Hideyuki; Yamamoto, Kyosuke; Yoshioka, Hideyoshi; Suzuki, Yuichiro; Kamagata, Yoichi; Sakata, Susumu

    2016-10-01

    Coal-bed methane is one of the largest unconventional natural gas resources. Although microbial activity may greatly contribute to coal-bed methane formation, it is unclear whether the complex aromatic organic compounds present in coal can be used for methanogenesis. We show that deep subsurface–derived Methermicoccus methanogens can produce methane from more than 30 types of methoxylated aromatic compounds (MACs) as well as from coals containing MACs. In contrast to known methanogenesis pathways involving one- and two-carbon compounds, this “methoxydotrophic” mode of methanogenesis couples O-demethylation, CO2 reduction, and possibly acetyl–coenzyme A metabolism. Because MACs derived from lignin may occur widely in subsurface sediments, methoxydotrophic methanogenesis would play an important role in the formation of natural gas not limited to coal-bed methane and in the global carbon cycle.

  10. Microbial community signature of high-solid content methanogenic ecosystems.

    PubMed

    Abbassi-Guendouz, Amel; Trably, Eric; Hamelin, Jérôme; Dumas, Claire; Steyer, Jean Philippe; Delgenès, Jean-Philippe; Escudié, Renaud

    2013-04-01

    In this study, changes in bacterial and archaeal communities involved in anaerobic digestion processes operated with high solid contents were investigated. Batch tests were performed within a range of total solids (TS) of 10-35%. Between 10% and 25% TS, high methanogenic activity was observed and no overall specific structure of active bacterial communities was found. At 30% and 35%, methanogenesis was inhibited as a consequence of volatile fatty acids accumulation. Here, a specific bacterial signature was observed with three main dominant bacteria related to Clostridium sp., known for their ability to grow at low pH. Additionally, archaeal community was gradually impacted by TS content. Three archaeal community structures were observed with a gradual shift from Methanobacterium sp. to Methanosarcina sp., according to the TS content. Overall, several species were identified as biomarkers of methanogenesis inhibition, since bacterial and archaeal communities were highly specific at high TS contents.

  11. Methanogenic burst in the end-Permian carbon cycle

    NASA Astrophysics Data System (ADS)

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

    2014-04-01

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

  12. Methanogenic burst in the end-Permian carbon cycle.

    PubMed

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

    2014-04-15

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

  13. [Analysis of methanogenic community of anaerobic granular sludge based on mcrA gene].

    PubMed

    Liu, Chu; Li, Liang; Ma, Jun-Ke; Wu, Gen; Yang, Jing-Liang

    2011-04-01

    The methanogenic community in anaerobic granular sludge from a full-scale UASB treating avernectin wastewater was analyzed based on mcrA gene, compared to 16S rRNA gene. The results indicated that the diversity indices of methanogenic community, including Shannon diversity index, Margalef richness index and Berger-Parker dominance index, were no difference between mcrA gene-based and 16S rRNA gene-based PCR products analysis by DGGE, although their DGGE band patterns were different, implying that the diversity analysis of methanogenic community based on mcrA genes was consistent with 16S rRNA gene. The phylogenetic analysis of dominant methanogenic populations based on these two target genes also showed resemble and Methanobacteriales and Methanosarcinales were determined to be the main orders of methanogenic populations in anaerobic granular sludge. On the other hand, the difference in phylogenetic analysis suggested simultaneously some group-specific of the two target genes. The hybridization of methanogenic community in FISH analysis based on two target genes was almost identical except a little different hybridization areas. The average relative abundance of methanogenic community was 24.25% +/- 6. 47% detected by FISH based on mcrA gene, lower than that based on 16S rRNA gene (33.42% +/- 2.34%). Then it could be concluded that the analysis of methanogenic community based on mcrA gene and 16S rRNA gene exhibited high resemblance and mcrA gene could used to be target gene for methanogenic community, as an alternative of 16S rRNA gene.

  14. Enhanced Biogas Production from Nanoscale Zero Valent Iron-Amended Anaerobic Bioreactors

    PubMed Central

    Carpenter, Alexis Wells; Laughton, Stephanie N.; Wiesner, Mark R.

    2015-01-01

    Abstract Addition of nanoscale zero valent iron (NZVI) to anaerobic batch reactors to enhance methanogenic activity is described. Two NZVI systems were tested: a commercially available NZVI (cNZVI) slurry and a freshly synthesized NZVI (sNZVI) suspension that was prepared immediately before addition to the reactors. In both systems, the addition of NZVI increased pH and decreased oxidation/reduction potential compared with unamended control reactors. Biodegradation of a model brewery wastewater was enhanced as indicated by an increase in chemical oxygen demand removal with both sNZVI and cNZVI amendments at all concentrations tested (1.25–5.0 g Fe/L). Methane production increased for all NZVI-amended bioreactors, with a maximum increase of 28% achieved on the addition of 2.5 and 5.0 g/L cNZVI. Addition of bulk zero-valent iron resulted in only a 5% increase in methane, indicating the advantage of using the nanoscale particles. NZVI amendments further improved produced biogas by decreasing the amount of CO2 released from the bioreactor by approximately 58%. Overall, addition of cNZVI proved more beneficial than the sNZVI at equal iron concentrations, due to decreased colloidal stability and larger effective particle size of sNZVI. Although some have reported cytotoxicity of NZVI to anaerobic microorganisms, work presented here suggests that NZVI of a certain particle size and reactivity can serve as an amendment to anaerobic digesters to enhance degradation and increase the value of the produced biogas, yielding a more energy-efficient anaerobic method for wastewater treatment. PMID:26339183

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

    USGS Publications Warehouse

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

    2005-01-01

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

  16. Methanogenesis and methanogenic pathways in a peat from subarctic permafrost.

    PubMed

    Metje, Martina; Frenzel, Peter

    2007-04-01

    Few studies have dealt so far with methanogenic pathways and populations in subarctic and arctic soils. We studied the effects of temperature on rates and pathways of CH4 production and on the relative abundance and structure of the archaeal community in a mildly acidic peat from a permafrost region in Siberia (67 degrees N). We monitored the production of CH4 and CO2 over time and measured the consumption of Fe(II), ethanol and volatile fatty acids. All experiments were performed with and without specific inhibitors [2-bromoethanesulfonate (BES) for methanogenesis and CH3F for acetoclastic methanogenesis]. The optimum temperature for methanogenesis was between 26 degrees C and 28 degrees C [4.3 micromol CH4 (g dry weight)(-1) day(-1)], but the activity was high even at 4 degrees C [0.75 micromol CH4 (g dry weight)(-1) day(-1)], constituting 17% of that at 27 degrees C. The population structure of archaea was studied by terminal restriction fragment length polymorphism analysis and remained constant over a wide temperature range. Acetoclastic methanogenesis accounted for about 70% of the total methanogenesis. Most 16S rRNA gene sequences clustered with Methanosarcinales, correlating with the prevalence of acetoclastic methanogenesis. In addition, sequences clustering with Methanobacteriales were recovered. Fe reduction occurred in parallel to methanogenesis. At lower and higher temperatures Fe reduction was not affected by BES. Because butyrate was consumed during methanogenesis and accumulated when methanogenesis was inhibited (BES and CH3F), it is proposed to serve as methanogenic precursor, providing acetate and H2 by syntrophic oxidation. In addition, ethanol and caproate occurred as intermediates. Because of thermodynamic constraints, homoacetogenesis could not compete with hydrogenotrophic methanogenesis. PMID:17359267

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

    PubMed

    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.

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

    SciTech Connect

    Roberts, M.F.

    1993-01-01

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

  19. 7 CFR 984.15 - Pack.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... Orders; Fruits, Vegetables, Nuts), DEPARTMENT OF AGRICULTURE WALNUTS GROWN IN CALIFORNIA Order Regulating Handling Definitions § 984.15 Pack. Pack means to bleach, clean, grade, shell or otherwise prepare walnuts for market as inshell or shelled walnuts....

  20. 7 CFR 984.15 - Pack.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... ORDERS; FRUITS, VEGETABLES, NUTS), DEPARTMENT OF AGRICULTURE WALNUTS GROWN IN CALIFORNIA Order Regulating Handling Definitions § 984.15 Pack. Pack means to bleach, clean, grade, shell or otherwise prepare walnuts for market as inshell or shelled walnuts....

  1. 7 CFR 984.15 - Pack.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... Orders; Fruits, Vegetables, Nuts), DEPARTMENT OF AGRICULTURE WALNUTS GROWN IN CALIFORNIA Order Regulating Handling Definitions § 984.15 Pack. Pack means to bleach, clean, grade, shell or otherwise prepare walnuts for market as inshell or shelled walnuts....

  2. 7 CFR 984.15 - Pack.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... ORDERS; FRUITS, VEGETABLES, NUTS), DEPARTMENT OF AGRICULTURE WALNUTS GROWN IN CALIFORNIA Order Regulating Handling Definitions § 984.15 Pack. Pack means to bleach, clean, grade, shell or otherwise prepare walnuts for market as inshell or shelled walnuts....

  3. 7 CFR 984.15 - Pack.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... Orders; Fruits, Vegetables, Nuts), DEPARTMENT OF AGRICULTURE WALNUTS GROWN IN CALIFORNIA Order Regulating Handling Definitions § 984.15 Pack. Pack means to bleach, clean, grade, shell or otherwise prepare walnuts for market as inshell or shelled walnuts....

  4. Designing gravel pack for uranium ISL wells

    NASA Astrophysics Data System (ADS)

    Ber, A. A.; Minaev, K. M.; Ber, L. M.; Isaev, Ye D.; Ulyanova, O. S.

    2016-09-01

    The paper describes the improvement of gravel packing technique applied for the production wells. The authors have suggested new design of gravel pack for gravel packing of productive formations. The issue is currently topical because gravel packing at drillhole ISL is less time- and money-consuming. The subject of the research is gravel pack design and content. The purpose defined by the authors is to design the gravel pack and to suggest the composition of gravel cement agent. As a result of the research, the authors have described different designs of the gravel pack, its optimal shape, as well as a choice and justification of cement agents, a hold cover of the gravel pack, and suggested the methods of experimental research.

  5. 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.

    PubMed

    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.

  6. Establishing Liver Bioreactors for In Vitro Research.

    PubMed

    Rebelo, Sofia P; Costa, Rita; Sousa, Marcos F Q; Brito, Catarina; Alves, Paula M

    2015-01-01

    In vitro systems that can effectively model liver function for long periods of time are fundamental tools for preclinical research. Nevertheless, the adoption of in vitro research tools at the earliest stages of drug development has been hampered by the lack of culture systems that offer the robustness, scalability, and flexibility necessary to meet industry's demands. Bioreactor-based technologies, such as stirred tank bioreactors, constitute a feasible approach to aggregate hepatic cells and maintain long-term three-dimensional cultures. These three-dimensional cultures sustain the polarity, differentiated phenotype, and metabolic performance of human hepatocytes. Culture in computer-controlled stirred tank bioreactors allows the maintenance of physiological conditions, such as pH, dissolved oxygen, and temperature, with minimal fluctuations. Moreover, by operating in perfusion mode, gradients of soluble factors and metabolic by-products can be established, aiming at resembling the in vivo microenvironment. This chapter provides a protocol for the aggregation and culture of hepatocyte spheroids in stirred tank bioreactors by applying perfusion mode for the long-term culture of human hepatocytes. This in vitro culture system is compatible with feeding high-throughput screening platforms for the assessment of drug elimination pathways, being a useful tool for toxicology research and drug development in the preclinical phase.

  7. Engineering stem cell niches in bioreactors

    PubMed Central

    Liu, Meimei; Liu, Ning; Zang, Ru; Li, Yan; Yang, Shang-Tian

    2013-01-01

    Stem cells, including embryonic stem cells, induced pluripotent stem cells, mesenchymal stem cells and amniotic fluid stem cells have the potential to be expanded and differentiated into various cell types in the body. Efficient differentiation of stem cells with the desired tissue-specific function is critical for stem cell-based cell therapy, tissue engineering, drug discovery and disease modeling. Bioreactors provide a great platform to regulate the stem cell microenvironment, known as “niches”, to impact stem cell fate decision. The niche factors include the regulatory factors such as oxygen, extracellular matrix (synthetic and decellularized), paracrine/autocrine signaling and physical forces (i.e., mechanical force, electrical force and flow shear). The use of novel bioreactors with precise control and recapitulation of niche factors through modulating reactor operation parameters can enable efficient stem cell expansion and differentiation. Recently, the development of microfluidic devices and microbioreactors also provides powerful tools to manipulate the stem cell microenvironment by adjusting flow rate and cytokine gradients. In general, bioreactor engineering can be used to better modulate stem cell niches critical for stem cell expansion, differentiation and applications as novel cell-based biomedicines. This paper reviews important factors that can be more precisely controlled in bioreactors and their effects on stem cell engineering. PMID:24179601

  8. MONITORING APPROACHES FOR BIOREACTOR LANDFILLS - Report

    EPA Science Inventory

    Experimental bioreactor landfill operations at operating Municipal Solid Waste (MSW) landfills can be approved under the research development and demonstration (RD&D) provisions of 30CFR 258.4. To provide a basis for consistent data collection for future decision-making in suppor...

  9. LANDFILL BIOREACTOR PERFORMANCE, SECOND INTERIM REPORT

    EPA Science Inventory

    A bioreactor landfill is a landfill that is operated in a manner that is expected to increase the rate and extent of waste decomposition, gas generation, and settlement compared to a traditional landfill. This Second Interim Report was prepared to provide an interpretation of fie...

  10. Continuous-Flow Gas-Phase Bioreactors

    NASA Technical Reports Server (NTRS)

    Wise, Donald L.; Trantolo, Debra J.

    1994-01-01

    Continuous-flow gas-phase bioreactors proposed for biochemical, food-processing, and related industries. Reactor contains one or more selected enzymes dehydrated or otherwise immobilized on solid carrier. Selected reactant gases fed into reactor, wherein chemical reactions catalyzed by enzyme(s) yield product biochemicals. Concept based on discovery that enzymes not necessarily placed in traditional aqueous environments to function as biocatalysts.

  11. Human cell culture in a space bioreactor

    NASA Technical Reports Server (NTRS)

    Morrison, Dennis R.

    1988-01-01

    Microgravity offers new ways of handling fluids, gases, and growing mammalian cells in efficient suspension cultures. In 1976 bioreactor engineers designed a system using a cylindrical reactor vessel in which the cells and medium are slowly mixed. The reaction chamber is interchangeable and can be used for several types of cell cultures. NASA has methodically developed unique suspension type cell and recovery apparatus culture systems for bioprocess technology experiments and production of biological products in microgravity. The first Space Bioreactor was designed for microprocessor control, no gaseous headspace, circulation and resupply of culture medium, and slow mixing in very low shear regimes. Various ground based bioreactors are being used to test reactor vessel design, on-line sensors, effects of shear, nutrient supply, and waste removal from continuous culture of human cells attached to microcarriers. The small Bioreactor is being constructed for flight experiments in the Shuttle Middeck to verify systems operation under microgravity conditions and to measure the efficiencies of mass transport, gas transfer, oxygen consumption and control of low shear stress on cells.

  12. In Vivo Bioreactors for Mandibular Reconstruction

    PubMed Central

    Tatara, A.M.; Wong, M.E.; Mikos, A.G.

    2014-01-01

    Large mandibular defects are difficult to reconstruct with good functional and aesthetic outcomes because of the complex geometry of craniofacial bone. While the current gold standard is free tissue flap transfer, this treatment is limited in fidelity by the shape of the harvested tissue and can result in significant donor site morbidity. To address these problems, in vivo bioreactors have been explored as an approach to generate autologous prefabricated tissue flaps. These bioreactors are implanted in an ectopic site in the body, where ossified tissue grows into the bioreactor in predefined geometries and local vessels are recruited to vascularize the developing construct. The prefabricated flap can then be harvested with vessels and transferred to a mandibular defect for optimal reconstruction. The objective of this review article is to introduce the concept of the in vivo bioreactor, describe important preclinical models in the field, summarize the human cases that have been reported through this strategy, and offer future directions for this exciting approach. PMID:25139360

  13. Denitrifying bioreactor clogging potential during wastewater treatment

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Chemoheterotrophic denitrification technologies using woodchips as a solid carbon source (i.e., woodchip bioreactors) have been widely trialed for treatment of diffuse-source agricultural nitrogen pollution. There is growing interest in the use of this simple, relatively low-cost biological wastewat...

  14. Evaluation of system performance and microbial communities of a bioaugmented anaerobic membrane bioreactor treating pharmaceutical wastewater.

    PubMed

    Ng, Kok Kwang; Shi, Xueqing; Ng, How Yong

    2015-09-15

    In this study, a control anaerobic membrane bioreactor (C-AnMBR) and a bioaugmented anaerobic membrane bioreactor (B-AnMBR) were operated for 210 d to treat pharmaceutical wastewater. Both the bioreactors were fed with the pharmaceutical wastewater containing TCOD of 16,249 ± 714 mg/L and total dissolved solids (TDS) of 29,450 ± 2209 mg/L with an organic loading rate (OLR) of 13.0 ± 0.6 kgCOD/m(3)d. Under steady-state condition, an average total chemical oxygen demand (TCOD) removal efficiency of 46.1 ± 2.9% and 60.3 ± 2.8% was achieved by the C-AnMBR and the B-AnMBR, respectively. The conventional anaerobes in the C-AnMBR cannot tolerate the hypersaline conditions well, resulting in lower TCOD removal efficiency, biogas production and methane yield than the B-AnMBR seeded from the coastal shore. Pyrosequencing analysis indicated that marine bacterial species (Oliephilus sp.) and halophilic bacterial species (Thermohalobacter sp.) were only present in the B-AnMBR; these species could possibly degrade complex and recalcitrant organic matter and withstand hypersaline environments. Two different dominant archaeal communities, genus Methanosaeta (43.4%) and Methanolobus (61.7%), were identified as the dominant methanogens in the C-AnMBR and the B-AnMBR, respectively. The species of genus Methanolobus was reported resistant to penicillin and required sodium and magnesium for growth, which could enable it to thrive in the hypersaline environment.

  15. 7 CFR 51.1527 - Standard pack.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ..., CERTIFICATION, AND STANDARDS) United States Standards for Grades of Fresh Plums and Prunes Standard Pack § 51... the approved and recognized methods. (2) The plums or prunes in the top layer of any package shall be...” means that the plums or prunes packed in loose or volume filled containers are packed within 1 inch...

  16. 7 CFR 51.1527 - Standard pack.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ..., CERTIFICATION, AND STANDARDS) United States Standards for Grades of Fresh Plums and Prunes Standard Pack § 51... the approved and recognized methods. (2) The plums or prunes in the top layer of any package shall be...” means that the plums or prunes packed in loose or volume filled containers are packed within 1 inch...

  17. 7 CFR 966.11 - Pack.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... ORDERS; FRUITS, VEGETABLES, NUTS), DEPARTMENT OF AGRICULTURE TOMATOES GROWN IN FLORIDA Order Regulating Handling Definitions § 966.11 Pack. Pack means any of the packs of tomatoes as defined and set forth in the United States Standards for Fresh Tomatoes issued by the United States Department of Agriculture (§§...

  18. 7 CFR 966.11 - Pack.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... ORDERS; FRUITS, VEGETABLES, NUTS), DEPARTMENT OF AGRICULTURE TOMATOES GROWN IN FLORIDA Order Regulating Handling Definitions § 966.11 Pack. Pack means any of the packs of tomatoes as defined and set forth in the United States Standards for Fresh Tomatoes issued by the United States Department of Agriculture (§§...

  19. 7 CFR 966.11 - Pack.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... Orders; Fruits, Vegetables, Nuts), DEPARTMENT OF AGRICULTURE TOMATOES GROWN IN FLORIDA Order Regulating Handling Definitions § 966.11 Pack. Pack means any of the packs of tomatoes as defined and set forth in the United States Standards for Fresh Tomatoes issued by the United States Department of Agriculture (§§...

  20. 7 CFR 966.11 - Pack.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... Orders; Fruits, Vegetables, Nuts), DEPARTMENT OF AGRICULTURE TOMATOES GROWN IN FLORIDA Order Regulating Handling Definitions § 966.11 Pack. Pack means any of the packs of tomatoes as defined and set forth in the United States Standards for Fresh Tomatoes issued by the United States Department of Agriculture (§§...

  1. 7 CFR 966.11 - Pack.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... Orders; Fruits, Vegetables, Nuts), DEPARTMENT OF AGRICULTURE TOMATOES GROWN IN FLORIDA Order Regulating Handling Definitions § 966.11 Pack. Pack means any of the packs of tomatoes as defined and set forth in the United States Standards for Fresh Tomatoes issued by the United States Department of Agriculture (§§...

  2. 7 CFR 51.1527 - Standard pack.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... package. The number of plums or prunes in California peach boxes or lug boxes shall not vary more than 4... Standards for Grades of Fresh Plums and Prunes Standard Pack § 51.1527 Standard pack. (a) Packing. (1) All... plums or prunes in the top layer of any package shall be reasonably representative in quality and...

  3. Diffusion in Jammed Particle Packs.

    PubMed

    Bolintineanu, Dan S; Grest, Gary S; Lechman, Jeremy B; Silbert, Leonardo E

    2015-08-21

    Using random walk simulations we explore diffusive transport through monodisperse sphere packings over a range of packing fractions ϕ in the vicinity of the jamming transition at ϕ(c). Various diffusion properties are computed over several orders of magnitude in both time and packing pressure. Two well-separated regimes of normal "Fickian" diffusion, where the mean squared displacement is linear in time, are observed. The first corresponds to diffusion inside individual spheres, while the latter is the long-time bulk diffusion. The intermediate anomalous diffusion regime and the long-time value of the diffusion coefficient are both shown to be controlled by particle contacts, which in turn depend on proximity to ϕ(c). The time required to recover normal diffusion t* scales as (ϕ-ϕ(c))(-0.5) and the long-time diffusivity D(∞)∼(ϕ-ϕ(c))0.5, or D(∞)∼1/t*. It is shown that the distribution of mean first passage times associated with the escape of random walkers between neighboring particles controls both t* and D(∞) in the limit ϕ→ϕ(c).

  4. Polyolefin Blend Miscibility and Packing

    NASA Astrophysics Data System (ADS)

    Lohse, David J.

    2000-03-01

    Over the last several years data have been obtained on the miscibility of a wide range of polyolefins, covering some 200 blends involving about 75 different components. Despite the fact that there are no 'specific interactions' between these saturated hydrocarbon polymers, every kind of phase behavior has been observed, including UCST, LCST, and even negative values of the Flory interaction parameter. The key factor that determines how these polyolefins mix is the way that they pack. Very often, polyolefins mix regularly, that is, the interaction energy is determined by the cohesive energies of the pure components. When they do mix regularly, miscibility is achieved by a close match in the packing lengths of the components. Favorable irregular mixing appears to be the result of some specific packing arrangements. Recent data on the effects of pressure and temperature on the mixing of several polyolefin blends shows that the interaction energies depend only on density (and not on T and P independently) for UCST blends far from a critical point. As a result, the effects of pressure on miscibility can be predicted for such blends from knowledge of the effects of temperature on the interactions combined with PVT data. This remarkable simplification appears to be related to the van der Waals nature of the interactions between saturated hydrocarbons. Density dependence predicts the trends correctly for LCST polyolefin blends, but for these mixtures the interactions depend in a more complex way on T and P.

  5. Diffusion in Jammed Particle Packs.

    PubMed

    Bolintineanu, Dan S; Grest, Gary S; Lechman, Jeremy B; Silbert, Leonardo E

    2015-08-21

    Using random walk simulations we explore diffusive transport through monodisperse sphere packings over a range of packing fractions ϕ in the vicinity of the jamming transition at ϕ(c). Various diffusion properties are computed over several orders of magnitude in both time and packing pressure. Two well-separated regimes of normal "Fickian" diffusion, where the mean squared displacement is linear in time, are observed. The first corresponds to diffusion inside individual spheres, while the latter is the long-time bulk diffusion. The intermediate anomalous diffusion regime and the long-time value of the diffusion coefficient are both shown to be controlled by particle contacts, which in turn depend on proximity to ϕ(c). The time required to recover normal diffusion t* scales as (ϕ-ϕ(c))(-0.5) and the long-time diffusivity D(∞)∼(ϕ-ϕ(c))0.5, or D(∞)∼1/t*. It is shown that the distribution of mean first passage times associated with the escape of random walkers between neighboring particles controls both t* and D(∞) in the limit ϕ→ϕ(c). PMID:26340211

  6. Bioreactors in tissue engineering - principles, applications and commercial constraints.

    PubMed

    Hansmann, Jan; Groeber, Florian; Kahlig, Alexander; Kleinhans, Claudia; Walles, Heike

    2013-03-01

    Bioreactor technology is vital for tissue engineering. Usually, bioreactors are used to provide a tissue-specific physiological in vitro environment during tissue maturation. In addition to this most obvious application, bioreactors have the potential to improve the efficiency of the overall tissue-engineering concept. To date, a variety of bioreactor systems for tissue-specific applications have been developed. Of these, some systems are already commercially available. With bioreactor technology, various functional tissues of different types were generated and cultured in vitro. Nevertheless, these efforts and achievements alone have not yet led to many clinically successful tissue-engineered implants. We review possible applications for bioreactor systems within a tissue-engineering process and present basic principles and requirements for bioreactor development. Moreover, the use of bioreactor systems for the expansion of clinically relevant cell types is addressed. In contrast to cell expansion, for the generation of functional three-dimensional tissue equivalents, additional physical cues must be provided. Therefore, bioreactors for musculoskeletal tissue engineering are discussed. Finally, bioreactor technology is reviewed in the context of commercial constraints.

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

    PubMed

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

    2011-06-01

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

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

    NASA Astrophysics Data System (ADS)

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

    2015-12-01

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

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

    EPA Science Inventory

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

  10. Functionally distinct genes regulated by hydrogen limitation and growth rate in methanogenic Archaea

    PubMed Central

    Hendrickson, Erik L.; Haydock, Andrew K.; Moore, Brian C.; Whitman, William B.; Leigh, John A.

    2007-01-01

    The use of molecular hydrogen as electron donor for energy generation is a defining characteristic of the hydrogenotrophic methanogens, an ancient group that dominates the phylum Eury archaeota. We present here a global study of changes in mRNA abundance in response to hydrogen availability for a hydrogenotrophic methanogen. Cells of Methanococcus maripaludis were grown by using continuous culture to deconvolute the effects of hydrogen limitation and growth rate, and microarray analyses were conducted. Hydrogen limitation markedly increased mRNA levels for genes encoding enzymes of the methanogenic pathway that reduce or oxidize the electron-carrying deazaflavin, coenzyme F420. F420-dependent redox functions in energy-generating metabolism are characteristic of the methanogenic Archaea, and the results show that their regulation is distinct from other redox processes in the cell. Rapid growth increased mRNA levels of the gene for an unusual hydrogenase, the hydrogen-dependent methylenetetrahydromethanopterin dehydrogenase. PMID:17502615

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

    PubMed Central

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

    2015-01-01

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

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

    NASA Astrophysics Data System (ADS)

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

    2011-06-01

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

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

    PubMed

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

    2015-01-01

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

  14. Relation between methanogenic archaea and methane production potential in selected natural wetland ecosystems across China

    NASA Astrophysics Data System (ADS)

    Liu, D. Y.; Ding, W. X.; Jia, Z. J.; Cai, Z. C.

    2011-02-01

    Methane (CH4) emissions from natural wetland ecosystems exhibit large spatial variability at regional, national, and global levels related to temperature, water table, plant type and methanogenic archaea etc. To understand the underlying factors that induce spatial differences in CH4 emissions, and the relationship between the population of methanogenic archaea and CH4 production potential in natural wetlands around China, we measured the CH4 production potential and the abundance of methanogenic archaea in vertical soil profiles sampled from the Poyang wetland in the subtropical zone, the Hongze wetland in the warm temperate zone, the Sanjiang marsh in the cold temperate zone, and the Ruoergai peatland in the Qinghai-Tibetan Plateau in the alpine climate zone. The top soil layer had the highest population of methanogens (1.07-8.29 × 109 cells g-1 soil) in all wetlands except the Ruoergai peatland and exhibited the maximum CH4 production potential measured at the mean in situ summer temperature. There is a significant logarithmic correlation between the abundance of methanogenic archaea and the soil organic carbon (R2 = 0.72, P < 0.001, n = 13) and between the abundance of methanogenic archaea and the total nitrogen concentrations (R2 = 0.76, P < 0.001, n = 13) in wetland soils. This indicates that the amount of soil organic carbon may affect the population of methanogens in wetland ecosystems. While the CH4 production potential is not significantly related to methanogen population (R2 = 0.01, P > 0.05, n = 13), it is related to the dissolved organic carbon concentration (R2 = 0.31, P = 0.05, n = 13). This suggests that the methanogen population might be not an effective index for predicting the CH4 production in wetland ecosystems. The CH4 production rate of the top soil layer increases with increasing latitude, from 273.64 μg CH4 kg-1 soil d-1 in the Poyang wetland to 664.59 μg CH4 kg-1 soil d-1 in the Carex lasiocarpa marsh of the Sanjiang Plain. We conclude

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

    PubMed Central

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

    2013-01-01

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

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

    PubMed

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

    2013-01-01

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

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

    PubMed

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

    2013-01-01

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

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

    PubMed

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

    2016-01-01

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

  19. Comparative study of fermentation and methanogen community structure in the digestive tract of goats and rabbits.

    PubMed

    Abecia, L; Fondevila, M; Rodríguez-Romero, N; Martínez, G; Yáñez-Ruiz, D R

    2013-05-01

    Methane is the most important anthropogenic contribution to climate change after carbon dioxide and represents a loss of feed energy for the animal, mainly for herbivorous species. However, our knowledge about the ecology of Archaea, the microbial group responsible for methane synthesis in the gut, is very poor. Moreover, it is well known that hindgut fermentation differs from rumen fermentation. The composition of archaeal communities in fermentation compartments of goats and rabbits were investigated using DGGE to generate fingerprints of archaeal 16S rRNA gene. Ruminal contents and faeces from five Murciano-Granadina goats and caecal contents of five commercial White New Zealand rabbits were compared. Diversity profile of methanogenic archaea was carried out by PCR-DGGE. Quantification of methanogenic archaea and the abundance relative to bacteria was determined by real-time PCR. Methanogenic archaeal species were relatively constant across species. Dendrogram from DGGE of the methanogen community showed one cluster for goat samples with two sub-clusters by type of sample (ruminal and faeces). In a second cluster, samples from rabbit were grouped. No differences were found either in richness or Shannon index as diversity indexes. Although the primer sets used was developed to investigate rumen methanogenic archaeal community, primers specificity did not affect the assessment of rabbit methanogen community structure. Rumen content showed the highest number or methanogenic archaea (log₁₀ 9.36), followed by faeces (log₁₀ 8.52) and showing rabbit caecum the lower values (log₁₀ 5.52). DGGE profile showed that pre-gastric and hindgut fermenters hold a very different methanogen community. Rabbits hold a microbial community of similar complexity than that in ruminants but less abundant, which agrees with the type of fermentation profile.

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

    NASA Astrophysics Data System (ADS)

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

    2007-12-01

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

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

    NASA Astrophysics Data System (ADS)

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

    2016-01-01

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

  2. Evidence of novel phylogenetic lineages of methanogenic archaea from hypersaline microbial mats.

    PubMed

    García-Maldonado, José Q; Bebout, Brad M; Everroad, R Craig; López-Cortés, Alejandro

    2015-01-01

    Methanogenesis in hypersaline and high-sulfate environments is typically dominated by methylotrophic methanogens because sulfate reduction is thermodynamically favored over hydrogenotrophic methanogenesis in these environments. We characterized the community composition of methanogenic archaea in both unmanipulated and incubated microbial mats from different hypersaline environments in Baja California Sur, Mexico. Clone libraries of methyl coenzyme-M reductase (mcrA) sequences and DGGE band patterns of 16S rRNA and mcrA sequences showed that the methanogen community in these microbial mats is dominated by methylotrophic methanogens of the genus Methanohalophilus. However, phylogenetic analyses of mcrA sequences from these mats also revealed two new lineages corresponding to putative hydrogenotrophic methanogens related with the strictly hydrogenotrophic order Methanomicrobiales. Stimulated methane production under decreased salinity and sulfate concentrations also suggested the presence of hydrogenotrophic methanogens in these samples. The relative abundance of mcrA gene and transcripts, estimated by SYBR green I qPCR assays, suggested the activity of different phylogenetic groups of methanogens, including the two novel clusters, in unmanipulated samples of hypersaline microbial mats. Using geochemical and molecular approaches, we show that substrate limitation and values of salinity and sulfate higher than 3 % and 25 mM (respectively) are potential environmental constraints for methanogenesis in these environments. Microcosm experiments with modifications of salinity and sulfate concentrations and TMA addition showed that upper salt and sulfate concentrations for occurrence of methylotrophic methanogenesis were 28 % and 263 mM, respectively. This study provides phylogenetic information about uncultivated and undescribed methanogenic archaea from hypersaline environments.

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

    PubMed

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

    2016-06-01

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

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

    NASA Astrophysics Data System (ADS)

    Ma, H.

    2014-12-01

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

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

    PubMed

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

    2015-08-01

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

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

    PubMed Central

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

    2015-01-01

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

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

    PubMed Central

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

    2016-01-01

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

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

    PubMed

    Krakat, Niclas; Schmidt, Stefan; Scherer, Paul

    2010-09-01

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

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

    PubMed

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

    2015-08-01

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

  10. Adhesive loose packings of small dry particles

    NASA Astrophysics Data System (ADS)

    Liu, Wenwei; Li, Shuiqing; Baule, Adrian; Makse, Hernán A.

    We explore adhesive loose packings of dry small spherical particles of micrometer size using 3D discrete-element simulations with adhesive contact mechanics. A dimensionless adhesion parameter ($Ad$) successfully combines the effects of particle velocities, sizes and the work of adhesion, identifying a universal regime of adhesive packings for $Ad>1$. The structural properties of the packings in this regime are well described by an ensemble approach based on a coarse-grained volume function that includes correlations between bulk and contact spheres. Our theoretical and numerical results predict: (i) An equation of state for adhesive loose packings that appears as a continuation from the frictionless random close packing (RCP) point in the jamming phase diagram; (ii) The existence of a maximal loose packing point at the coordination number $Z=2$ and packing fraction $\\phi=1/2^{3}$. Our results highlight that adhesion leads to a universal packing regime at packing fractions much smaller than the random loose packing, which can be described within a statistical mechanical framework. We present a general phase diagram of jammed matter comprising frictionless, frictional, adhesive as well as non-spherical particles, providing a classification of packings in terms of their continuation from the spherical frictionless RCP.

  11. Kinetics of pack aluminization of nickel

    NASA Technical Reports Server (NTRS)

    Seigle, L. L.; Gupta, B. K.; Shankar, R.; Sarkhel, A. K.

    1978-01-01

    The kinetics of pack aluminization of unalloyed nickel in packs of varying aluminum activity with various halide activators were studied. Surface compositions of the coatings as functions of time, temperature, and pack composition were obtained in order to establish the boundary conditions for diffusion in the system. The structure of the packs was also examined in order to clarify the mechanism of aluminum transport. The results indicate that the kinetics of pack aluminization are controlled jointly by gas diffusion in the pack and solid diffusion in the coating. Levine and Caves' model for gas diffusion was combined with calculations of rates of diffusion in the solid to formulate a more complete theory for the kinetics of pack aluminization.

  12. High-rate, High Temperature Acetotrophic Methanogenesis Governed by a Three Population Consortium in Anaerobic Bioreactors

    PubMed Central

    Ho, Dang; Jensen, Paul; Gutierrez-Zamora, Maria-Luisa; Beckmann, Sabrina; Manefield, Mike; Batstone, Damien

    2016-01-01

    A combination of acetate oxidation and acetoclastic methanogenesis has been previously identified to enable high-rate methanogenesis at high temperatures (55 to 65°C), but this capability had not been linked to any key organisms. This study combined RNA–stable isotope probing on 13C-labelled acetate and 16S amplicon sequencing to identify the active micro-organisms involved in high-rate methanogenesis. Active biomass was harvested from three bench-scale thermophilic bioreactors treating waste activated sludge at 55, 60 and 65°C, and fed with 13-C labelled and 12C-unlabelled acetate. Acetate uptake and cumulative methane production were determined and kinetic parameters were estimated using model-based analysis. Pyrosequencing performed on 13C- enriched samples indicated that organisms accumulating labelled carbon were Coprothermobacter (all temperatures between 55 and 65°C), acetoclastic Methanosarcina (55 to 60°C) and hydrogenotrophic Methanothermobacter (60 to 65°C). The increased relative abundance of Coprothermobacter with increased temperature corresponding with a shift to syntrophic acetate oxidation identified this as a potentially key oxidiser. Methanosarcina likely acts as both a hydrogen utilising and acetoclastic methanogen at 55°C, and is replaced by Methanothermobacter as a hydrogen utiliser at higher temperatures. PMID:27490246

  13. High-rate, High Temperature Acetotrophic Methanogenesis Governed by a Three Population Consortium in Anaerobic Bioreactors.

    PubMed

    Ho, Dang; Jensen, Paul; Gutierrez-Zamora, Maria-Luisa; Beckmann, Sabrina; Manefield, Mike; Batstone, Damien

    2016-01-01

    A combination of acetate oxidation and acetoclastic methanogenesis has been previously identified to enable high-rate methanogenesis at high temperatures (55 to 65°C), but this capability had not been linked to any key organisms. This study combined RNA-stable isotope probing on 13C-labelled acetate and 16S amplicon sequencing to identify the active micro-organisms involved in high-rate methanogenesis. Active biomass was harvested from three bench-scale thermophilic bioreactors treating waste activated sludge at 55, 60 and 65°C, and fed with 13-C labelled and 12C-unlabelled acetate. Acetate uptake and cumulative methane production were determined and kinetic parameters were estimated using model-based analysis. Pyrosequencing performed on 13C- enriched samples indicated that organisms accumulating labelled carbon were Coprothermobacter (all temperatures between 55 and 65°C), acetoclastic Methanosarcina (55 to 60°C) and hydrogenotrophic Methanothermobacter (60 to 65°C). The increased relative abundance of Coprothermobacter with increased temperature corresponding with a shift to syntrophic acetate oxidation identified this as a potentially key oxidiser. Methanosarcina likely acts as both a hydrogen utilising and acetoclastic methanogen at 55°C, and is replaced by Methanothermobacter as a hydrogen utiliser at higher temperatures. PMID:27490246

  14. Iron deficiency and bioavailability in anaerobic batch and submerged membrane bioreactors (SAMBR) during organic shock loads.

    PubMed

    Ketheesan, Balachandran; Thanh, Pham Minh; Stuckey, David C

    2016-07-01

    This study examined the effects of Fe(2+) and its bioavailability for controlling VFAs during organic shock loads in batch reactors and a submerged anaerobic membrane bioreactor (SAMBR). When seed grown under Fe-sufficient conditions (7.95±0.05mgFe/g-TSS), an organic shock resulted in leaching of Fe from the residual to organically bound and soluble forms. Under Fe-deficient seed conditions (0.1±0.002mgFe/gTSS), Fe(2+) supplementation (3.34mgFe(2+)/g-TSS) with acetate resulted in a 2.1-3.9 fold increase in the rate of methane production, while with propionate it increased by 1.2-1.5 fold compared to non-Fe(2+) supplemented reactors. Precipitation of Fe(2+) as sulphides and organically bound Fe were bioavailable to methanogens for acetate assimilation. The results confirmed that the transitory/long term limitations of Fe play a significant role in controlling the degradation of VFAs during organic shock loads due to their varying physical/chemical states, and bioavailability. PMID:27015020

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

    PubMed

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

    2015-06-01

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

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

    PubMed

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

    2012-12-01

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

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

    PubMed

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

    2015-06-01

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

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

    PubMed

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

    2015-04-01

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

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

    PubMed Central

    Angel, Roey; Claus, Peter; Conrad, Ralf

    2012-01-01

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

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

    USGS Publications Warehouse

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

    1984-01-01

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

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

    NASA Astrophysics Data System (ADS)

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

    2015-12-01

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

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

    SciTech Connect

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

    2014-10-22

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

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

    DOE PAGES

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

    2014-10-22

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

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

    PubMed Central

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

    2015-01-01

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

  5. Methanogenic Oil Degradation in the Dagang Oil Field

    NASA Astrophysics Data System (ADS)

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

    2014-05-01

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

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

    NASA Astrophysics Data System (ADS)

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

    2015-01-01

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

  7. Bioreactor Development for Lung Tissue Engineering

    PubMed Central

    Panoskaltsis-Mortari, Angela

    2015-01-01

    Rationale Much recent interest in lung bioengineering by pulmonary investigators, industry and the organ transplant field has seen a rapid growth of bioreactor development ranging from the microfluidic scale to the human-sized whole lung systems. A comprehension of the findings from these models is needed to provide the basis for further bioreactor development. Objective The goal was to comprehensively review the current state of bioreactor development for the lung. Methods A search using PubMed was done for published, peer-reviewed papers using the keywords “lung” AND “bioreactor” or “bioengineering” or “tissue engineering” or “ex vivo perfusion”. Main Results Many new bioreactors ranging from the microfluidic scale to the human-sized whole lung systems have been developed by both academic and commercial entities. Microfluidic, lung-mimic and lung slice cultures have the advantages of cost-efficiency and high throughput analyses ideal for pharmaceutical and toxicity studies. Perfused/ventilated rodent whole lung systems can be adapted for mid-throughput studies of lung stem/progenitor cell development, cell behavior, understanding and treating lung injury and for preliminary work that can be translated to human lung bioengineering. Human-sized ex vivo whole lung bioreactors incorporating perfusion and ventilation are amenable to automation and have been used for whole lung decellularization and recellularization. Clinical scale ex vivo lung perfusion systems have been developed for lung preservation and reconditioning and are currently being evaluated in clinical trials. Conclusions Significant advances in bioreactors for lung engineering have been made at both the microfluidic and the macro scale. The most advanced are closed systems that incorporate pressure-controlled perfusion and ventilation and are amenable to automation. Ex vivo lung perfusion systems have advanced to clinical trials for lung preservation and reconditioning. The biggest

  8. Hollow fiber bioreactor technology for tissue engineering applications.

    PubMed

    Eghbali, Hadis; Nava, Michele M; Mohebbi-Kalhori, Davod; Raimondi, Manuela T

    2016-01-01

    Hollow fiber bioreactors are the focus of scientific research aiming to mimic physiological vascular networks and engineer organs and tissues in vitro. The reason for this lies in the interesting features of this bioreactor type, including excellent mass transport properties. Indeed, hollow fiber bioreactors allow limitations to be overcome in nutrient transport by diffusion, which is often an obstacle to engineer sizable constructs in vitro. This work reviews the existing literature relevant to hollow fiber bioreactors in organ and tissue engineering applications. To this purpose, we first classify the hollow fiber bioreactors into 2 categories: cylindrical and rectangular. For each category, we summarize their main applications both at the tissue and at the organ level, focusing on experimental models and computational studies as predictive tools for designing innovative, dynamic culture systems. Finally, we discuss future perspectives on hollow fiber bioreactors as in vitro models for tissue and organ engineering applications.

  9. Cultivating microbial dark matter in benzene-degrading methanogenic consortia.

    PubMed

    Luo, Fei; Devine, Cheryl E; Edwards, Elizabeth A

    2016-09-01

    The microbes responsible for anaerobic benzene biodegradation remain poorly characterized. In this study, we identified and quantified microbial populations in a series of 16 distinct methanogenic, benzene-degrading enrichment cultures using a combination of traditional 16S rRNA clone libraries (four cultures), pyrotag 16S rRNA amplicon sequencing (11 cultures), metagenome sequencing (1 culture) and quantitative polymerase chain reaction (qPCR; 12 cultures). An operational taxonomic unit (OTU) from the Deltaproteobacteria designated ORM2 that is only 84% to 86% similar to Syntrophus or Desulfobacterium spp. was consistently identified in all enrichment cultures, and typically comprised more than half of the bacterial sequences. In addition to ORM2, a sequence belonging to Parcubacteria (candidate division OD1) identified from the metagenome data was the only other OTU common to all the cultures surveyed. Culture transfers (1% and 0.1%) were made in the presence and absence of benzene, and the abundance of ORM2, OD1 and other OTUs was tracked over 415 days using qPCR. ORM2 sequence abundance increased only when benzene was present, while the abundance of OD1 and other OTUs increased even in the absence of benzene. Deltaproteobacterium ORM2 is unequivocally the benzene-metabolizing population. This study also hints at laboratory cultivation conditions for a member of the widely distributed yet uncultivated Parcubacteria (OD1).

  10. Isolation and Characterization of an H2-Oxidizing Thermophilic Methanogen

    PubMed Central

    Ferguson, Thomas J.; Mah, Robert A.

    1983-01-01

    A thermophilic methanogen was isolated from enrichment cultures originally inoculated with sludge from an anaerobic kelp digester (55°C). This isolate exhibited a temperature optimum of 55 to 60°C and a maximum near 70°C. Growth occurred throughout the pH range of 5.5 to 9.0, with optimal growth near pH 7.2. Although 4% salt was present in the isolation medium, salt was not required for optimal growth. The thermophile utilized formate or H2-CO2 but not acetate, methanol, or methylamines for growth and methanogenesis. Growth in complex medium was very rapid, and a minimum doubling time of 1.8 h was recorded in media supplemented with rumen fluid. Growth in defined media required the addition of acetate and an unknown factor(s) from digester supernatant, rumen fluid, or Trypticase. Cells in liquid culture were oval to coccoid, 0.7 to 1.8 μm in diameter, often occurring in pairs. The cells were easily lysed upon exposure to oxygen or 0.08 mg of sodium dodecyl sulfate per ml. The isolate was sensitive to tetracycline and chloramphenicol but not penicillin G or cycloserine. The DNA base composition was 59.69 mol% guanine plus cytosine. Images PMID:16346171

  11. Cultivating microbial dark matter in benzene-degrading methanogenic consortia.

    PubMed

    Luo, Fei; Devine, Cheryl E; Edwards, Elizabeth A

    2016-09-01

    The microbes responsible for anaerobic benzene biodegradation remain poorly characterized. In this study, we identified and quantified microbial populations in a series of 16 distinct methanogenic, benzene-degrading enrichment cultures using a combination of traditional 16S rRNA clone libraries (four cultures), pyrotag 16S rRNA amplicon sequencing (11 cultures), metagenome sequencing (1 culture) and quantitative polymerase chain reaction (qPCR; 12 cultures). An operational taxonomic unit (OTU) from the Deltaproteobacteria designated ORM2 that is only 84% to 86% similar to Syntrophus or Desulfobacterium spp. was consistently identified in all enrichment cultures, and typically comprised more than half of the bacterial sequences. In addition to ORM2, a sequence belonging to Parcubacteria (candidate division OD1) identified from the metagenome data was the only other OTU common to all the cultures surveyed. Culture transfers (1% and 0.1%) were made in the presence and absence of benzene, and the abundance of ORM2, OD1 and other OTUs was tracked over 415 days using qPCR. ORM2 sequence abundance increased only when benzene was present, while the abundance of OD1 and other OTUs increased even in the absence of benzene. Deltaproteobacterium ORM2 is unequivocally the benzene-metabolizing population. This study also hints at laboratory cultivation conditions for a member of the widely distributed yet uncultivated Parcubacteria (OD1). PMID:26549712

  12. Molecular characterization of a toluene-degrading methanogenic consortium

    SciTech Connect

    Ficker, M.; Krastel, K.; Orlicky, S.; Edwards, E.

    1999-12-01

    A toluene-degrading methanogenic consortium enriched from creosote-contaminated aquifer material was maintained on toluene as the sole carbon and energy source for 10 years. The species in the consortium were characterized by using a molecular approach. Total genomic DNA was isolated, and 16S rRBA genes were amplified by using PCR performed with kingdom-specific primers that were specific for 16S rRBA genes from either members of the kingdom Bacteria or members of the kingdom Archaea. A total of 90 eubacterial clones and 75 archaeal clones were grouped by performing a restriction fragment length polymorphism (RFLP) analysis. Six eubacterial sequences and two archaeal sequences were found in the greatest abundance (in six or more clones) based on the RFLP analysis. The relative abundance of each putative species was estimated by using fluorescent in situ hybridization (FISH), and the presence of putative species was determined qualitatively by performing slot blot hybridization with consortium DNA. Both archael species and two of the six eubacterial species were detected in the DNA and FISH hybridization experiments. A phylogenetic analysis of these four dominant organisms suggested that the two archaeal species are related to the genera methanosaeta and Methanospirillum. One of the eubacterial species is related to the genus Desulfotomaculum, which the others is not related to any previously described genus. By elimination, the authors propose that the last organism probably initiates the attack on toluene.

  13. Functional responses of methanogenic archaea to syntrophic growth

    PubMed Central

    Walker, Christopher B; Redding-Johanson, Alyssa M; Baidoo, Edward E; Rajeev, Lara; He, Zhili; Hendrickson, Erik L; Joachimiak, Marcin P; Stolyar, Sergey; Arkin, Adam P; Leigh, John A; Zhou, Jizhong; Keasling, Jay D; Mukhopadhyay, Aindrila; Stahl, David A

    2012-01-01

    Methanococcus maripaludis grown syntrophically with Desulfovibrio vulgaris was compared with M. maripaludis monocultures grown under hydrogen limitation using transcriptional, proteomic and metabolite analyses. These measurements indicate a decrease in transcript abundance for energy-consuming biosynthetic functions in syntrophically grown M. maripaludis, with an increase in transcript abundance for genes involved in the energy-generating central pathway for methanogenesis. Compared with growth in monoculture under hydrogen limitation, the response of paralogous genes, such as those coding for hydrogenases, often diverged, with transcripts of one variant increasing in relative abundance, whereas the other was little changed or significantly decreased in abundance. A common theme was an apparent increase in transcripts for functions using H2 directly as reductant, versus those using the reduced deazaflavin (coenzyme F420). The greater importance of direct reduction by H2 was supported by improved syntrophic growth of a deletion mutant in an F420-dependent dehydrogenase of M. maripaludis. These data suggest that paralogous genes enable the methanogen to adapt to changing substrate availability, sustaining it under environmental conditions that are often near the thermodynamic threshold for growth. Additionally, the discovery of interspecies alanine transfer adds another metabolic dimension to this environmentally relevant mutualism. PMID:22739494

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

    SciTech Connect

    Banaszak, J. E.

    1998-12-21

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

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

    SciTech Connect

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

    1982-01-01

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

  16. Fate of neptunium in an anaerobic, methanogenic microcosm

    SciTech Connect

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

    1999-07-01

    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{sup {minus}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.

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

    PubMed

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

    2015-07-01

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

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

    PubMed

    Jones, J G; Simon, B M

    1985-04-01

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

  19. Advanced bioreactors for enhanced production of chemicals

    SciTech Connect

    Davison, B.H.

    1993-12-31

    A variety of advanced bioreactors are being developed to improve production of fuels, solvents, organic acids, and other fermentation products. One key approach is immobilization of the biocatalyst, leading to increased rates and yields. In addition, there are processes for simultaneous fermentation and separation to further increase production. For example, ethanol productivity in immobilized-cell fluidized-bed bioreactors (FBRs) can increase more than tenfold with 99% conversion and near stoichiometric yields. Two modified FBR configurations offer further improvements by removing the inhibitory product directly from the continuous fermentation. One involves the addition and removal of solid adsorbent particles to the FBR. This process was demonstrated with the production of lactic acid by immobilized Lactobacillus. The second uses an immiscible organic extractant in the FBR. This increased total butanol yields in the anaerobic acetone-butanol fermentation by Clostridium acetobutylicum.

  20. The role of bioreactors in tissue engineering.

    PubMed

    Martin, Ivan; Wendt, David; Heberer, Michael

    2004-02-01

    Ex vivo engineering of living tissues is a rapidly developing area with the potential to impact significantly on a wide-range of biomedical applications. Major obstacles to the generation of functional tissues and their widespread clinical use are related to a limited understanding of the regulatory role of specific physicochemical culture parameters on tissue development, and the high manufacturing costs of the few commercially available engineered tissue products. By enabling reproducible and controlled changes of specific environmental factors, bioreactor systems provide both the technological means to reveal fundamental mechanisms of cell function in a 3D environment, and the potential to improve the quality of engineered tissues. In addition, by automating and standardizing tissue manufacture in controlled closed systems, bioreactors could reduce production costs, thus facilitating a wider use of engineered tissues.

  1. Bioreactor-Based Tumor Tissue Engineering

    PubMed Central

    Guller, A.E.; Grebenyuk, P.N.; Shekhter, A.B.; Zvyagin, A.V.; Deyev, S. M.

    2016-01-01

    This review focuses on modeling of cancer tumors using tissue engineering technology. Tumor tissue engineering (TTE) is a new method of three-dimensional (3D) simulation of malignant neoplasms. Design and development of complex tissue engineering constructs (TECs) that include cancer cells, cell-bearing scaffolds acting as the extracellular matrix, and other components of the tumor microenvironment is at the core of this approach. Although TECs can be transplanted into laboratory animals, the specific aim of TTE is the most realistic reproduction and long-term maintenance of the simulated tumor properties in vitro for cancer biology research and for the development of new methods of diagnosis and treatment of malignant neoplasms. Successful implementation of this challenging idea depends on bioreactor technology, which will enable optimization of culture conditions and control of tumor TECs development. In this review, we analyze the most popular bioreactor types in TTE and the emerging applications. PMID:27795843

  2. Chromium detoxification by fixed-film bioreactors

    SciTech Connect

    Chirwa, E.M.N.; Wang, Y.T.

    1996-11-01

    In this study, completely mixed, continuous flow bioreactors were utilized to detoxify chromium. Glass beads were incorporated as a support medium for two strains of bacteria, Bacillus sp. and Pseudomonas fluorescens LB300 (LB300), growing aerobically in two separate reactors. Aerobic conditions were maintained in the reactors by continuously supplying fresh air to the liquid through gas exchange chambers installed on the recycle line of the bioreactors. Results obtained showed that near complete removal of chromate was possible for influent concentrations up to 200 mg/L for Bacillus sp., and up to 100 mg/L for LB300 at 24 hours liquid detention time. Similar results were obtained for corresponding loading rates at 12 hours and 6 hours liquid detention time.

  3. Advanced bioreactors for enhanced production of chemicals

    SciTech Connect

    Davison, B.H.; Scott, C.D.

    1993-06-01

    A variety of advanced bioreactors are being developed to improve production of fuels, solvents, organic acids and other fermentation products. One key approach is immobilization of the biocatalyst leading to increased rates and yields. In addition, there are processes for simultaneous fermentation and separation to further increase production by the removal of an inhibitory product. For example, ethanol productivity in immobilized-cell fluidized-bed bioreactors (FBRs) can increase more than tenfold with 99% conversion and near stoichiometric yields. Two modified FBR configurations offer further improvements by removing the inhibitory product directly from the continuous fermentation. One involves the addition and removal of solid adsorbent particles to the FBR. This process was demonstrated with the production of lactic acid by immobilized Lactobacillus. The second uses an immiscible organic extractant in the FBR. This increased total butanol yields in the anaerobic acetone-butanol fermentation by Clostridium acetobutylicum.

  4. Bioreactor and process design for biohydrogen production.

    PubMed

    Show, Kuan-Yeow; Lee, Duu-Jong; Chang, Jo-Shu

    2011-09-01

    Biohydrogen is regarded as an attractive future clean energy carrier due to its high energy content and environmental-friendly conversion. It has the potential for renewable biofuel to replace current hydrogen production which rely heavily on fossil fuels. While biohydrogen production is still in the early stage of development, there have been a variety of laboratory- and pilot-scale systems developed with promising potential. This work presents a review of advances in bioreactor and bioprocess design for biohydrogen production. The state-of-the art of biohydrogen production is discussed emphasizing on production pathways, factors affecting biohydrogen production, as well as bioreactor configuration and operation. Challenges and prospects of biohydrogen production are also outlined.

  5. Bioreactor and process design for biohydrogen production.

    PubMed

    Show, Kuan-Yeow; Lee, Duu-Jong; Chang, Jo-Shu

    2011-09-01

    Biohydrogen is regarded as an attractive future clean energy carrier due to its high energy content and environmental-friendly conversion. It has the potential for renewable biofuel to replace current hydrogen production which rely heavily on fossil fuels. While biohydrogen production is still in the early stage of development, there have been a variety of laboratory- and pilot-scale systems developed with promising potential. This work presents a review of advances in bioreactor and bioprocess design for biohydrogen production. The state-of-the art of biohydrogen production is discussed emphasizing on production pathways, factors affecting biohydrogen production, as well as bioreactor configuration and operation. Challenges and prospects of biohydrogen production are also outlined. PMID:21624834

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

    PubMed

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

    2016-05-01

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

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

    PubMed

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

    2016-05-01

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

  8. Sulfur-based mixotrophic denitrification corresponding to different electron donors and microbial profiling in anoxic fluidized-bed membrane bioreactors.

    PubMed

    Zhang, Lili; Zhang, Chao; Hu, Chengzhi; Liu, Huijuan; Bai, Yaohui; Qu, Jiuhui

    2015-11-15

    Sulfur-based mixotrophic denitrifying anoxic fluidized bed membrane bioreactors (AnFB-MBR) were developed for the treatment of nitrate-contaminated groundwater with minimized sulfate production. The nitrate removal rates obtained in the methanol- and ethanol-fed mixotrophic denitrifying AnFB-MBRs reached 1.44-3.84 g NO3 -N/L reactor d at a hydraulic retention time of 0.5 h, which were significantly superior to those reported in packed bed reactors. Compared to methanol, ethanol was found to be a more effective external carbon source for sulfur-based mixotrophic denitrification due to lower sulfate and total organic carbon concentrations in the effluent. Using pyrosequencing, the phylotypes of primary microbial groups in the reactor, including sulfur-oxidizing autotrophic denitrifiers, methanol- or ethanol-supported heterotrophic denitrifiers, were investigated in response to changes in electron donors. Principal component and heatmap analyses indicated that selection of electron donating substrates largely determined the microbial community structure. The abundance of Thiobacillus decreased from 45.1% in the sulfur-oxidizing autotrophic denitrifying reactor to 12.0% and 14.2% in sulfur-based methanol- and ethanol-fed mixotrophic denitrifying bioreactors, respectively. Heterotrophic Methyloversatilis and Thauera bacteria became more dominant in the mixotrophic denitrifying bioreactors, which were possibly responsible for the observed methanol- and ethanol-associated denitrification.

  9. Design concepts for bioreactors in space

    NASA Technical Reports Server (NTRS)

    Seshan, P. K.; Peterson, G. R.; Beard, B.; Boshe, C.; Dunlop, E. H.

    1987-01-01

    Microbial food sources are becoming viable and more efficient alternatives to conventional food sources, especially in the context of closed ecological life support systems (CELSS) in space habitats. Two bioreactor design concepts presented represent two dissimilar approaches to grappling with the absence of gravity in space habitats and deserve to be tested for adoption as important components of the life support function aboard spacecraft, space stations and other extra-terrestrial habitats.

  10. Oxygen transfer in a pressurized airlift bioreactor.

    PubMed

    Campani, Gilson; Ribeiro, Marcelo Perencin Arruda; Horta, Antônio Carlos Luperni; Giordano, Roberto Campos; Badino, Alberto Colli; Zangirolami, Teresa Cristina

    2015-08-01

    Airlift bioreactors (ALBs) offer advantages over conventional systems, such as simplicity of construction, reduced risk of contamination, and efficient gas-liquid dispersion with low power consumption. ALBs are usually operated under atmospheric pressure. However, in bioprocesses with high oxygen demand, such as high cell density cultures, oxygen limitation may occur even when operating with high superficial gas velocity and air enriched with oxygen. One way of overcoming this drawback is to pressurize the reactor. In this configuration, it is important to assess the influence of bioreactor internal pressure on the gas hold-up, volumetric oxygen transfer coefficient (k(L)a), and volumetric oxygen transfer rate (OTR). Experiments were carried out in a concentric-tube airlift bioreactor with a 5 dm(3) working volume, equipped with a system for automatic monitoring and control of the pressure, temperature, and inlet gas flow rate. The results showed that, in disagreement with previous published results for bubble column and external loop airlift reactors, overpressure did not significantly affect k(L)a within the studied ranges of pressure (0.1-0.4 MPa) and superficial gas velocity in the riser (0.032-0.065 m s(-1)). Nevertheless, a positive effect on OTR was observed: it increased up to 5.4 times, surpassing by 2.3 times the oxygen transfer in a 4 dm(3) stirred tank reactor operated under standard cultivation conditions. These results contribute to the development of non-conventional reactors, especially pneumatic bioreactors operated using novel strategies for oxygen control.

  11. Bioreactor Yields Extracts for Skin Cream

    NASA Technical Reports Server (NTRS)

    2015-01-01

    Johnson Space Flight Center researchers created a unique rotating-wall bioreactor that simulates microgravity conditions, spurring innovations in drug development and medical research. Renuèll Int'l Inc., based in Aventure, Florida, licensed the technology and used it to produce a healing skin care product, RE`JUVEL. In a Food and Drug Administration test, RE`JUVEL substantially increased skin moisture and elasticity while reducing dark blotches and wrinkles.

  12. Enhanced biodegradation of Reactive Violet 5R manufacturing wastewater using down flow fixed film bioreactor.

    PubMed

    Sheth, Niraj; Dave, Shailesh

    2010-11-01

    The present study emphasizes on the development of bioprocess for biodegradation and bioremediation of Reactive Violet 5R (RV5) manufacturing industry's wastewater in laboratory scale indigenously designed down flow fixed film bioreactor (DFFR). Process was investigated in DFFR, packed with furnace charcoal as a support material. During the batch and continuous operation of DFFR more than 95% degradation, 88% COD reduction and 99% copper remediation was obtained in less than 8h of contact time. Continuous mode treatment gave degradation of more than 2500mg dye in only 1h of contact time. Addition of 0.25% peptone enhanced biodegradation rate more than three-fold. The biologically treated wastewater was found to stimulate the growth of selected soil bacteria. Degradation of major components of the wastewater was confirmed by HPTLC. DFFR was operated successfully for 750d continuously, during which 716 cycles of batch treatment were operated.

  13. Diffusion in jammed particle packs

    NASA Astrophysics Data System (ADS)

    Bolintineanu, Dan S.; Silbert, Leonardo E.; Grest, Gary S.; Lechman, Jeremy B.

    2015-03-01

    Diffusive transport in jammed particle packs is of interest for a number of applications, as well as being a potential indicator of structural properties near the jamming point. To this end, we report stochastic simulations of equilibrium diffusion through monodisperse sphere packs near the jamming point in the limit of a perfectly insulating surrounding medium. The time dependence of various diffusion properties is resolved over several orders of magnitude. Two time regimes of expected Fickian diffusion are observed, separated by an intermediate regime of anomalous diffusion. This intermediate regime grows as the particle volume fraction approaches the critical jamming transition. The diffusion behavior is fully controlled by the extent of the contacts between neighboring particles, which in turn depend on proximity to the jamming point. In particular, the mean first passage time associated with the escape of random walkers between neighboring particles is shown to control both the time to recover Fickian diffusion and the long time diffusivity. Scaling laws are established that relate these quantities to the difference between the actual and critical jamming volume fractions. Sandia National Laboratories is a multiprogram laboratory managed and operated by Sandia Corporation, a wholly owned subsidiary of Lockheed Martin Corporation, for the U.S. Department of Energy's NNSA under Contract DE- AC04-94AL85000.

  14. A bioreactor system for the nitrogen loop in a controlled ecological life support system

    NASA Astrophysics Data System (ADS)

    Saulmon, M. M.; Reardon, K. F.; Sadeh, W. Z.

    1996-01-01

    As space missions become longer in duration, the need to recycle waste into useful compounds rises dramatically. This problem can be addressed by the development of Controlled Ecological Life Support Systems (CELSS) (i.e., Engineered Closed/ Controlled Eco-Systems (ECCES)), consisting of human and plant modules. One of the waste streams leaving the human module is urine. In addition to the reclamation of water from urine, recovery of the nitrogen is important because it is an esssential nutrient for the plant module. A 3-step biological process for the recycling of nitrogenous waste (urea) is proposed. A packed-bed bioreactor system for this purpose was modeled, and the issues of reaction step segregation, reactor type and volume, support particle size, and pressure drop were addressed. Based on minimization of volume, a bioreactor system consisting of a plug flow immobilized urease reactor, a completely mixed flow immobilized cell reactor to convert ammonia to nitrite, and a plug flow immobilized cell reactor to produce nitrate from nitrite is recommended. It is apparent that this 3-step bioprocess meets the requirements for space applications.

  15. A bioreactor system for the nitrogen loop in a Controlled Ecological Life Support System

    NASA Technical Reports Server (NTRS)

    Saulmon, M. M.; Reardon, K. F.; Sadeh, W. Z.

    1996-01-01

    As space missions become longer in duration, the need to recycle waste into useful compounds rises dramatically. This problem can be addressed by the development of Controlled Ecological Life Support Systems (CELSS) (i.e., Engineered Closed/Controlled Eco-Systems (ECCES)), consisting of human and plant modules. One of the waste streams leaving the human module is urine. In addition to the reclamation of water from urine, recovery of the nitrogen is important because it is an essential nutrient for the plant module. A 3-step biological process for the recycling of nitrogenous waste (urea) is proposed. A packed-bed bioreactor system for this purpose was modeled, and the issues of reaction step segregation, reactor type and volume, support particle size, and pressure drop were addressed. Based on minimization of volume, a bioreactor system consisting of a plug flow immobilized urease reactor, a completely mixed flow immobilized cell reactor to convert ammonia to nitrite, and a plug flow immobilized cell reactor to produce nitrate from nitrite is recommended. It is apparent that this 3-step bioprocess meets the requirements for space applications.

  16. Raman measurement of glucose in bioreactor materials

    NASA Astrophysics Data System (ADS)

    Xu, Yijing; Ford, Joseph F.; Mann, Charles K.; Vickers, Thomas J.; Brackett, John M.; Cousineau, Kristen L.; Robey, W. G.

    1997-06-01

    The feasibility of using Raman spectroscopy to monitor the concentration of chemical species in a bioreactor has been examined. Successful operation of a bioreactor requires that nutrients and metabolic waste products be maintained within narrow ranges, and it is, therefore, important to provide accurate, reliable and timely measurement of the composition in the reactor. Raman spectroscopy offers the possibility of real time simultaneous monitoring of molecular components present in the millimolar and higher concentration range. Work reported here has focused on four analytes: glucose, glutamine, lactic acid and ammonia. Measurements have been made with a spectrograph providing a spectral window for simultaneous measurement of about 1800 cm-1 on a multichannel CCD detector. Most measurements were made with an argon ion laser emitting at 514.5 nm. Some measurements are reported with a solid state diode laser operating at 785 nm. Locally constructed inexpensive silica fiber-optic probes delivered the laser light and collected the scattered radiation. Spectra of the four analytes n buffer and reactor media have been obtained. Analytical curves have been constructed and limits of detection measured. Limits of detection in buffer media are about 1 mM. Results are reported for off-line measurements on material drawn from a bioreactor.

  17. Replaceable Sensor System for Bioreactor Monitoring

    NASA Technical Reports Server (NTRS)

    Mayo, Mike; Savoy, Steve; Bruno, John

    2006-01-01

    A sensor system was proposed that would monitor spaceflight bioreactor parameters. Not only will this technology be invaluable in the space program for which it was developed, it will find applications in medical science and industrial laboratories as well. Using frequency-domain-based fluorescence lifetime technology, the sensor system will be able to detect changes in fluorescence lifetime quenching that results from displacement of fluorophorelabeled receptors bound to target ligands. This device will be used to monitor and regulate bioreactor parameters including glucose, pH, oxygen pressure (pO2), and carbon dioxide pressure (pCO2). Moreover, these biosensor fluorophore receptor-quenching complexes can be designed to further detect and monitor for potential biohazards, bioproducts, or bioimpurities. Biosensors used to detect biological fluid constituents have already been developed that employ a number of strategies, including invasive microelectrodes (e.g., dark electrodes), optical techniques including fluorescence, and membrane permeable systems based on osmotic pressure. Yet the longevity of any of these sensors does not meet the demands of extended use in spacecraft habitat or bioreactor monitoring. It was therefore necessary to develop a sensor platform that could determine not only fluid variables such as glucose concentration, pO2, pCO2, and pH but can also regulate these fluid variables with controlled feedback loop.

  18. The Packing of Granular Polymer Chains

    SciTech Connect

    Zou, Ling-Nan; Cheng, Xiang; Rivers, Mark L.; Jaeger, Heinrich M.; Nagel, Sidney R.; UC

    2009-12-01

    Rigid particles pack into structures, such as sand dunes on the beach, whose overall stability is determined by the average number of contacts between particles. However, when packing spatially extended objects with flexible shapes, additional concepts must be invoked to understand the stability of the resulting structure. Here, we examine the disordered packing of chains constructed out of flexibly connected hard spheres. Using x-ray tomography, we find that long chains pack into a low-density structure whose mechanical rigidity is mainly provided by the backbone. On compaction, randomly oriented, semi-rigid loops form along the chain, and the packing of chains can be understood as the jamming of these elements. Finally, we uncover close similarities between the packing of chains and the glass transition in polymers.

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

    PubMed

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

    2015-01-01

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

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

    PubMed

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

    2015-12-01

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