Sample records for anaerobic thermophiles annual

  1. Anaerobic thermophilic culture

    DOEpatents

    Ljungdahl, Lars G.; Wiegel, Jurgen K. W.

    1981-01-01

    A newly discovered thermophilic anaerobe is described that was isolated in a biologically pure culture and designated Thermoanaerobacter ethanolicus ATCC 3/550. T. Ethanolicus is cultured in aqueous nutrient medium under anaerobic, thermophilic conditions and is used in a novel process for producing ethanol by subjecting carbohydrates, particularly the saccharides, to fermentation action of the new microorganism in a biologically pure culture.

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

    PubMed

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

    2015-02-01

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

  3. Anaerobic Thermophiles

    PubMed Central

    Canganella, Francesco; Wiegel, Juergen

    2014-01-01

    The term “extremophile” was introduced to describe any organism capable of living and growing under extreme conditions. With the further development of studies on microbial ecology and taxonomy, a variety of “extreme” environments have been found and an increasing number of extremophiles are being described. Extremophiles have also been investigated as far as regarding the search for life on other planets and even evaluating the hypothesis that life on Earth originally came from space. The first extreme environments to be largely investigated were those characterized by elevated temperatures. The naturally “hot environments” on Earth range from solar heated surface soils and water with temperatures up to 65 °C, subterranean sites such as oil reserves and terrestrial geothermal with temperatures ranging from slightly above ambient to above 100 °C, to submarine hydrothermal systems with temperatures exceeding 300 °C. There are also human-made environments with elevated temperatures such as compost piles, slag heaps, industrial processes and water heaters. Thermophilic anaerobic microorganisms have been known for a long time, but scientists have often resisted the belief that some organisms do not only survive at high temperatures, but actually thrive under those hot conditions. They are perhaps one of the most interesting varieties of extremophilic organisms. These microorganisms can thrive at temperatures over 50 °C and, based on their optimal temperature, anaerobic thermophiles can be subdivided into three main groups: thermophiles with an optimal temperature between 50 °C and 64 °C and a maximum at 70 °C, extreme thermophiles with an optimal temperature between 65 °C and 80 °C, and finally hyperthermophiles with an optimal temperature above 80 °C and a maximum above 90 °C. The finding of novel extremely thermophilic and hyperthermophilic anaerobic bacteria in recent years, and the fact that a large fraction of them belong to the Archaea has

  4. Anaerobic thermophilic culture system

    DOEpatents

    Ljungdahl, Lars G.; Wiegel, Jurgen K. W.

    1981-01-01

    A mixed culture system of the newly discovered microorganism Thermoanaerobacter ethanolicus ATCC31550 and the microorganism Clostridium thermocellum ATCC31549 is described. In a mixed nutrient culture medium that contains cellulose, these microorganisms have been coupled and cultivated to efficiently ferment cellulose to produce recoverable quantities of ethanol under anaerobic, thermophilic conditions.

  5. Diversity of Cultured Thermophilic Anaerobes in Hot Springs of Yunnan Province, China

    NASA Astrophysics Data System (ADS)

    Lin, L.; Lu, Y.; Dong, X.; Liu, X.; Wei, Y.; Ji, X.; Zhang, C.

    2010-12-01

    Thermophilic anaerobes including Archaea and Bacteria refer to those growing optimally at temperatures above 50°C and do not use oxygen as the terminal electron acceptor for growth. Study on thermophilic anaerobes will help to understand how life thrives under extreme conditions. Meanwhile thermophilic anaerobes are of importance in potential application and development of thermophilic biotechnology. We have surveyed culturable thermophilic anaerobes in hot springs (pH6.5-7.5; 70 - 94°C) in Rehai of Tengchong, Bangnazhang of Longlin, Eryuan of Dali,Yunnan, China. 50 strains in total were cultured from the hot springs water using Hungate anaerobic technique, and 30 strains were selected based on phenotypic diversity for analysis of 16S rDNA sequences. Phylogenetic analysis showed that 28 strains belonged to the members of five genera: Caldanaerobacter, Calaramator, Thermoanaerobacter, Dictyoglomus and Fervidobacterium, which formed five branches on the phylogenetic tree. Besides, 2 strains of methanogenic archaea were obtained. The majority of the isolates were the known species, however, seven strains were identified as novel species affiliated to the five genera based on the lower 16S rDNA sequence similarities (less than 93 - 97%) with the described species. This work would provide the future study on their diversity, distribution among different regions and the potential application of thermophilic enzyme. Supported by State Key Laboratory of Microbial Resources, Institute of Microbiology, Chinese Academy of Sciences(SKLMR-080605)and the Foundation of State Natural Science (30660009, 30960022, 31081220175).

  6. Microthrix parvicella and Gordona amarae in mesophilic and thermophilic anaerobic digestion systems.

    PubMed

    Marneri, Matina; Mamais, Daniel; Koutsiouki, Efi

    2009-04-14

    The scope of the study presented in this paper is to determine the fate of the filamentous bacteria Gordona amarae and Microthrix parvicella in anaerobic digestion operating under mesophilic and thermophilic conditions. In order to detect and quantify foaming bacteria in the anaerobic digesters, a fluorescent in situ hybridization (FISH) method was developed and applied. This paper presents the results of a laboratory-scale study that involved the operation of four lab-scale anaerobic digestion systems operating in the mesophilic (35 degrees C) and thermophilic (55 degrees C) temperature ranges at 20 days' detention time. According to the FISH counts of G. amarae and M. parvicella, it appears that thermophilic conditions resulted in a higher destruction of both filamentous bacteria, averaging approximately 97% and 94% for the single thermophilic digester and the dual thermophilic/mesophilic system, respectively. Within the context of this study, the overall performance of the four different anaerobic digestion systems was evaluated in terms of biogas production per mass of volatile solids destroyed, COD destruction, sludge dewaterability and foaming characteristics. The dual stage systems used in this study outperformed the single stage digesters.

  7. Mesophilic and thermophilic anaerobic digestion of biologically pretreated abattoir wastewaters in an upflow anaerobic filter.

    PubMed

    Gannoun, H; Bouallagui, H; Okbi, A; Sayadi, S; Hamdi, M

    2009-10-15

    The hydrolysis pretreatment of abattoir wastewaters (AW), rich in organic suspended solids (fats and protein) was studied in static and stirred batch reactors without aeration in the presence of natural microbial population acclimated in a storage tank of AW. Microbial analysis showed that the major populations which contribute to the pretreatment of AW belong to the genera Bacillus. Contrary to the static pretreatment, the stirred conditions favoured the hydrolysis and solubilization of 80% of suspended matter into soluble pollution. The pretreated AW, in continuous stirred tank reactor (CSTR) at a hydraulic retention time (HRT) of 2 days, was fed to an upflow anaerobic filter (UAF) at an HRT of 2 days. The performance of anaerobic digestion of biologically pretreated AW was examined under mesophilic (37 degrees C) and thermophilic (55 degrees C) conditions. The shifting from a mesophilic to a thermophilic environment in the UAF was carried out with a short start-up of thermophilic condition. The UAF ran at organic loading rates (OLRs) ranging from 0.9 to 6g COD/Ld in mesophilic conditions and at OLRs from 0.9 to 9 g COD/Ld in thermophilic conditions. COD removal efficiencies of 80-90% were achieved for OLRs up to 4.5 g COD/Ld in mesophilic conditions, while the highest OLRs i.e. 9 g COD/Ld led to efficiencies of 70-72% in thermophilic conditions. The biogas yield in thermophilic conditions was about 0.32-0.45 L biogas/g of COD removed for OLRs up to 4.5 g COD/Ld. For similar OLR, the UAF in mesophilic conditions showed lower percentage of methanization. Mesophilic anaerobic digestion has been shown to destroy pathogens partially, whereas the thermophilic process was more efficient in the removal of indicator microorganisms and pathogenic bacteria at different organic loading rates.

  8. Behavior of cellulose-degrading bacteria in thermophilic anaerobic digestion process.

    PubMed

    Syutsubo, K; Nagaya, Y; Sakai, S; Miya, A

    2005-01-01

    Previously, we found that the newly isolated Clostridium sp. strain JC3 became the dominant cellulose-degrading bacterium in thermophilic methanogenic sludge. In the present study, the behavior of strain JC3 in the thermophilic anaerobic digestion process was investigated quantitatively by molecular biological techniques. A cellulose-degrading experiment was conducted at 55 degrees C with a 9.5 L of anaerobic baffled reactor having three compartments (Nos. 1, 2, 3). Over 80% of the COD input was converted into methane when 2.5 kgCOD m(-3) d(-1) was loaded for an HRT of 27 days. A FISH probe specific for strain JC3 was applied to sludge samples harvested from the baffled reactor. Consequently, the ratio of JC3 cells to DAPI-stained cells increased from below 0.5% (undetectable) to 9.4% (compartment 1), 13.1% (compartment 2) and 21.6% (compartment 3) at day 84 (2.5 kgCOD m(-3)d(-1)). The strain JC3 cell numbers determined by FISH correlated closely with the cellulose-degrading methanogenic activities of retained sludge. A specific primer set targeting the cellulase gene (cellobiohydrolaseA: cbhA) of strain JC3 was designed and applied to digested sludge for treating solid waste such as coffee grounds, wastepaper, garbage, cellulose and so on. The strain JC3 cell numbers determined by quantitative PCR correlated closely with the cellulose-sludge loading of the thermophilic digester. Strain JC3 is thus important in the anaerobic hydrolysis of cellulose in thermophilic anaerobic digestion processes.

  9. Hemicellulases from the ethanologenic thermophile, Thermoanaerobacter ethanolicus and related anaerobic thermophiles. Final report, September 1992--June 1996

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Wiegel, J.

    1998-09-01

    The short term goals of this application were to characterize hemicellulases from anaerobic thermophiles on the biochemical and molecular level to extend the presently limited knowledge of hemicellulases in anaerobic thermophilic bacteria. This objective includes the following tasks: (1) Traditional purification and biochemical/biophysical characterization of xylanases from the newly isolated, slightly alkalitolerant strain NDF190, and the slightly acid-tolerant strain YS485, both with high xylanolytic activities, and of the 4-O-methyl glucuronidase and arabinosidase from strain NDF190 and the acetyl (xylan) esterase from T. ethanolicus. This also includes determining the N-terminal sequences and obtaining gene probes. (2) Elucidation of the regulation ofmore » hemicellulolytic enzymes in anaerobic thermophiles. (3) To clone into E. coli and identify the multiplicity of the enzymes involved in hemicellulose degradation by T. ethanolicus and other suitable organisms. (4) To purify and characterize the recombinant enzymes with the goal of identifying the best enzymes for cloning into the ethanologenic T. ethanolicus to obtain an optimized hemicellulose utilization by this bacterium.« less

  10. Hemicellulases from the ethanologenic thermophile Thermoanaerobacter ethanolicus and related anaerobic thermophiles. Final report, September 1992--June 1996

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Wiegel, J.

    1998-05-01

    The SHORT TERM GOALS of this application were to characterize hemicellulases from anaerobic thermophiles on the biochemical and molecular level to extend the presently limited knowledge of hemicellulases in anaerobic thermophilic bacteria. This objective includes the following TASKS: (1) Traditional purification and biochemical/biophysical characterization of xylanases from the newly isolated, slightly alkalitolerant strain NDF190, and the slightly acid-tolerant strain YS485, both with high xylanolytic activities, and of the 4-0-methyl glucuronidase and arabinosidase from strain NDF190 and the acetyl (xylan) esterase from T. ethanolicus. This also includes determining the N-terminal sequences and obtaining gene probes. (2) Elucidation of the regulation ofmore » hemicellulolytic enzymes in anaerobic thermophiles. (3) To clone into E. coli and identify the multiplicity of the enzymes involved in hemicellulose degradation by T. ethanolicus and other suitable organisms. (4) To purify and characterize the recombinant enzymes with the goal of identifying the best enzymes for cloning into the ethanologenic T. ethanolicus to obtain an optimized hemicellulose utilization by this bacterium (one of our long term goals).« less

  11. Growth media in anaerobic fermentative processes: The underestimated potential of thermophilic fermentation and anaerobic digestion.

    PubMed

    Hendriks, A T W M; van Lier, J B; de Kreuk, M K

    Fermentation and anaerobic digestion of organic waste and wastewater is broadly studied and applied. Despite widely available results and data for these processes, comparison of the generated results in literature is difficult. Not only due to the used variety of process conditions, but also because of the many different growth media that are used. Composition of growth media can influence biogas production (rates) and lead to process instability during anaerobic digestion. To be able to compare results of the different studies reported, and to ensure nutrient limitation is not influencing observations ascribed to process dynamics and/or reaction kinetics, a standard protocol for creating a defined growth medium for anaerobic digestion and mixed culture fermentation is proposed. This paper explains the role(s) of the different macro- and micronutrients, as well as the choices for a growth medium formulation strategy. In addition, the differences in nutrient requirements between mesophilic and thermophilic systems are discussed as well as the importance of specific trace metals regarding specific conversion routes and the possible supplementary requirement of vitamins. The paper will also give some insight into the bio-availability and toxicity of trace metals. A remarkable finding is that mesophilic and thermophilic enzymes are quite comparable at their optimum temperatures. This has consequences for the trace metal requirements of thermophiles under certain conditions. Under non-limiting conditions, the trace metal requirement of thermophilic systems is about 3 times higher than for mesophilic systems. Copyright © 2017 Elsevier Inc. All rights reserved.

  12. Enhanced anaerobic degradability of highly polluted pesticides-bearing wastewater under thermophilic conditions.

    PubMed

    García-Mancha, N; Monsalvo, V M; Puyol, D; Rodriguez, J J; Mohedano, A F

    2017-10-05

    This work presents a sustainable and cost-competitive solution for hardly biodegradable pesticides-bearing wastewater treatment in an anaerobic expanded granular sludge bed (EGSB) reactor at mesophilic (35°C) and thermophilic (55°C). The reactor was operated in continuous mode during 160days, achieving an average COD removal of 33 and 44% under mesophilic and thermophilic conditions, respectively. The increase of temperature improved the biomass activity and the production of methane by 35%. Around 96% of pesticides identified in raw wastewater were not detected in both mesophilic and thermophilic effluents. A dramatic selection of the microbial population in anaerobic granules was caused by the presence of pesticides, which also changed significantly when the temperature was increased. Pesticides caused a significant inhibition on methanogenesis, especially over acetoclastic methanogens. Aerobic biodegradability tests of the resulting anaerobic effluents revealed that aerobic post-treatment is also a feasible and effective option, yielding more than 60% COD reduction. Copyright © 2017 Elsevier B.V. All rights reserved.

  13. Microbial diversity in innovative mesophilic/thermophilic temperature-phased anaerobic digestion of sludge.

    PubMed

    Gagliano, M C; Braguglia, C M; Gallipoli, A; Gianico, A; Rossetti, S

    2015-05-01

    Anaerobic digestion (AD) is one of the few sustainable technologies that both produce energy and treat waste streams. Driven by a complex and diverse community of microbes, AD may be affected by different factors, many of which also influence the composition and activity of the microbial community. In this study, the biodiversity of microbial populations in innovative mesophilic/thermophilic temperature-phased AD of sludge was evaluated by means of fluorescence in situ hybridization (FISH). The increase of digestion temperature drastically affected the microbial composition and selected specialized biomass. Hydrogenotrophic Methanobacteriales and the protein fermentative bacterium Coprothermobacter spp. were identified in the thermophilic anaerobic biomass. Shannon-Weaver diversity (H') and evenness (E) indices were calculated using FISH data. Species richness was lower under thermophilic conditions compared with the values estimated in mesophilic samples, and it was flanked by similar trend of the evenness indicating that thermophilic communities may be therefore more susceptible to sudden changes and less prompt to adapting to operative variations.

  14. State of the art and future perspectives of thermophilic anaerobic digestion.

    PubMed

    Ahring, B K; Mladenovska, Z; Iranpour, R; Westermann, P

    2002-01-01

    The state of the art of thermophilic digestion is discussed. Thermophilic digestion is a well established technology in Europe for treatment of mixtures of waste in common large scale biogas plants or for treatment of the organic fraction of municipal solid waste. Due to a large number of failures over time with thermophilic digestion of sewage sludge this process has lost its appeal in the USA. New demands on sanitation of biosolids before land use will, however, bring the attention back to the use of elevated temperatures during sludge stabilization. In the paper we show how the use of a start-up strategy based on the actual activity of key microbes can be used to ensure proper and fast transfer of mesophilic digesters into thermophilic operation. Extreme thermophilic temperatures of 65 degrees C or more may be necessary in the future to meet the demands for full sanitation of the waste material before final disposal. We show data of anaerobic digestion at extreme thermophilic temperatures.

  15. Mesophilic and thermophilic anaerobic digestion of sulphate-containing wastewaters.

    PubMed

    Colleran, E; Pender, S

    2002-01-01

    The effect of sulphate at an influent chemical oxygen demand (COD):sulphate ratio of 4 on the operational performance of anaerobic hybrid reactors treating molasses wastewater was investigated under mesophilic and thermophilic conditions in a long-term laboratory-scale study over a 1,081 day period. The presence of sulphate reduced the COD removal efficiency under both mesophilic and thermophilic conditions. At 55 degrees C, effluent acetate levels were consistently greater than 4000 mg l(-1) indicating that thermophilic acetate-utilising methane-producing bacteria (MPB) or sulphate-reducing bacteria (SRB) had not developed in the reactor under the conditions applied. At 37 degrees C, acetate was exclusively utilised by acetoclastic methanogens, whereas H2-utilising SRB predominated over H2-utilising MPB in the competition for hydrogen. By contrast, hydrogenotrophic MPB were shown to outcompete H2-utilising SRB during long-term thermophilic operation. 16SrDNA analysis of the seed sludge and reactor biomass on conclusion of the 37 degrees C and 55 degrees C trials illustrated that the dominant methanogen present on conclusion of the thermophilic trial in the absence of influent sulphate was related to Methanocorpusculum parvuum, and was capable of growth on both acetate and hydrogen. By contrast, an organism closely related to Methanobacterium thermoautotrophicum was the dominant methanogen present in the sulphate-fed reactor on completion of the thermophilic trial.

  16. In situ identification of the synthrophic protein fermentative Coprothermobacter spp. involved in the thermophilic anaerobic digestion process.

    PubMed

    Gagliano, Maria Cristina; Braguglia, Camilla Maria; Rossetti, Simona

    2014-09-01

    Thermophilic bacteria have recently attracted great attention because of their potential application in improving different biochemical processes such as anaerobic digestion of various substrates, wastewater treatment or hydrogen production. In this study we report on the design of a specific 16S rRNA-targeted oligonucleotide probe for detecting members of Coprothermobacter genus characterized by a strong protease activity to degrade proteins and peptides. The newly designed CTH485 probe and helper probes hCTH429 and hCTH439 were optimized for use in fluorescence in situ hybridization (FISH) on thermophilic anaerobic sludge samples. In situ probing revealed that thermo-adaptive mechanisms shaping the 16S rRNA gene may affect the identification of thermophilic microorganisms. The novel developed FISH probe extends the possibility to study the widespread thermophilic syntrophic interaction of Coprothermobacter spp. with hydrogenotrophic methanogenic archaea, whose establishment is a great benefit for the whole anaerobic system. © 2014 Federation of European Microbiological Societies. Published by John Wiley & Sons Ltd. All rights reserved.

  17. Influence of temperature fluctuation on thermophilic anaerobic digestion of municipal organic solid waste*

    PubMed Central

    Wu, Man-Chang; Sun, Ke-Wei; Zhang, Yong

    2006-01-01

    A laboratory-scale experiment was carried out to assess the influence of temperature fluctuation on thermophilic anaerobic digestion of municipal organic solid waste (MOSW). Heating failure was simulated by decreasing temperature suddenly from 55 °C to 20 °C suddenly; 2 h time is needed for temperature decrease and recovery. Under the conditions of 8.0 g/(L·d) and 15 d respectively for MOSW load and retention time, following results were noted: (1) biogas production almost stopped and VFA (volatile fatty acid) accumulated rapidly, accompanied by pH decrease; (2) with low temperature (20 °C) duration of 1, 5, 12 and 24 h, it took 3, 11, 56 and 72 h for the thermophilic anaerobic digestion system to reproduce methane after temperature fluctuation; (3) the longer the low temperature interval lasted, the more the methanogenic bacteria would decay; hydrolysis, acidification and methanogenesis were all influenced by temperature fluctuation; (4) the thermophilic microorganisms were highly resilient to temperature fluctuation. PMID:16502503

  18. Cellulosic ethanol production via consolidated bioprocessing by a novel thermophilic anaerobic bacterium isolated from a Himalayan hot spring.

    PubMed

    Singh, Nisha; Mathur, Anshu S; Tuli, Deepak K; Gupta, Ravi P; Barrow, Colin J; Puri, Munish

    2017-01-01

    Cellulose-degrading thermophilic anaerobic bacterium as a suitable host for consolidated bioprocessing (CBP) has been proposed as an economically suited platform for the production of second-generation biofuels. To recognize the overall objective of CBP, fermentation using co-culture of different cellulolytic and sugar-fermenting thermophilic anaerobic bacteria has been widely studied as an approach to achieving improved ethanol production. We assessed monoculture and co-culture fermentation of novel thermophilic anaerobic bacterium for ethanol production from real substrates under controlled conditions. In this study, Clostridium sp. DBT-IOC-C19, a cellulose-degrading thermophilic anaerobic bacterium, was isolated from the cellulolytic enrichment cultures obtained from a Himalayan hot spring. Strain DBT-IOC-C19 exhibited a broad substrate spectrum and presented single-step conversion of various cellulosic and hemicellulosic substrates to ethanol, acetate, and lactate with ethanol being the major fermentation product. Additionally, the effect of varying cellulose concentrations on the fermentation performance of the strain was studied, indicating a maximum cellulose utilization ability of 10 g L -1 cellulose. Avicel degradation kinetics of the strain DBT-IOC-C19 displayed 94.6% degradation at 5 g L -1 and 82.74% degradation at 10 g L -1 avicel concentration within 96 h of fermentation. In a comparative study with Clostridium thermocellum DSM 1313, the ethanol and total product concentrations were higher by the newly isolated strain on pretreated rice straw at an equivalent substrate loading. Three different co-culture combinations were used on various substrates that presented two-fold yield improvement than the monoculture during batch fermentation. This study demonstrated the direct fermentation ability of the novel thermophilic anaerobic bacteria on various cellulosic and hemicellulosic substrates into ethanol without the aid of any exogenous enzymes

  19. Previously unclassified bacteria dominate during thermophilic and mesophilic anaerobic pre-treatment of primary sludge.

    PubMed

    Pervin, Hasina M; Batstone, Damien J; Bond, Philip L

    2013-06-01

    Thermophilic biological pre-treatment enables enhanced anaerobic digestion for treatment of wastewater sludges but, at present, there is limited understanding of the hydrolytic-acidogenic microbial composition and its contribution to this process. In this study, the process was assessed by comparing the microbiology of thermophilic (50-65 °C) and mesophilic (35 °C) pre-treatment reactors treating primary sludge. A full-cycle approach for the 16S rRNA genes was applied in order to monitor the diversity of bacteria and their abundance in a thermophilic pre-treatment reactor treating primary sludge. For the thermophilic pre-treatment (TP), over 90% of the sequences were previously undetected and these had less than 97% sequence similarity to cultured organisms. During the first 83 days, members of the Betaproteobacteria dominated the community sequences and a newly designed probe was used to monitor a previously unknown bacterium affiliated with the genus Brachymonas. Between days 85 and 183, three phylotypes that affiliated with the genera Comamonas, Clostridium and Lysobacter were persistently dominant in the TP community, as revealed by terminal-restriction fragment length polymorphism (T-RFLP). Hydrolytic and fermentative functions have been speculated for these bacteria. Mesophilic pre-treatment (MP) and TP communities were different but they were both relatively dynamic. Statistical correlation analysis and the function of closely allied reference organisms indicated that previously unclassified bacteria dominated the TP community and may have been functionally involved in the enhanced hydrolytic performance of thermophilic anaerobic pre-treatment. This study is the first to reveal the diversity and dynamics of bacteria during anaerobic digestion of primary sludge. Copyright © 2013 Elsevier GmbH. All rights reserved.

  20. Degradation characteristics of polylactide in thermophilic anaerobic digestion with hyperthermophilic solubilization condition.

    PubMed

    Wang, F; Hidaka, T; Oishi, T; Osumi, S; Tsubota, J; Tsuno, H

    2011-01-01

    To test whether hyperthermophilic treatment promotes polylactide (PLA) dissolution and methane conversion under anaerobic digestion conditions, a single thermophilic control reactor (55 °C) and a two-phase system consisting of a hyperthermophilic reactor (80 °C) and a thermophilic reactor (55 °C) were continuously fed with a mixture of PLA and artificial kitchen garbage. In Runs 1 and 2, the PLA dissolution ratios in the two-phase system were 79.2 ± 6.5% and 85.2 ± 7.0%, respectively, higher than those of the control. Batch experimental results indicated that hyperthermophilic treatment could promote PLA dissolution to a greater degree as compared with single thermophilic treatment and that ammonia addition also had a promotional effect on PLA dissolution. In the two-phase system, after hyperthermophilic treatment, dissolved PLA was converted to methane gas under the subsequent thermophilic condition.

  1. Impact of ammonia and sulphate concentration on thermophilic anaerobic digestion.

    PubMed

    Siles, J A; Brekelmans, J; Martín, M A; Chica, A F; Martín, A

    2010-12-01

    The effect of increasing concentrations of ammonia and sulphate on thermophilic anaerobic digestion (52 degrees C) was studied at laboratory-scale. The substrate consisted of a synthetic solution supplemented with ammonia and sodium sulphate. In terms of biogas production, the results showed that the C/N and C/SO(4)(2-) thresholds were 4.40 and 1.60, respectively, corresponding to 620 mg FA (free ammonia)/L and 1400 mg SO(4)(2-)/L. No reduction in biogas production was observed until reaching the above concentration of sulphate in the sulphate toxicity test. However, when the concentration of ammonia was increased to 620 mg FA/L in the ammonia toxicity test, a gradual decrease of 21% was observed for the biogas. In order to characterise each set of experiments kinetically, a biogas production first-order kinetic model was used to fit the experimental data. The proposed model accurately predicted the behaviour of the microorganisms affecting the thermophilic anaerobic digestion, allowing its evolution to be predicted. 2010 Elsevier Ltd. All rights reserved.

  2. Efficiency of autothermal thermophilic aerobic digestion and thermophilic anaerobic digestion of municipal wastewater sludge in removing Salmonella spp. and indicator bacteria.

    PubMed

    Zábranská, J; Dohányos, M; Jenícek, P; Růziciková, H; Vránová, A

    2003-01-01

    The study is focused on the comparison of autothermal thermophilic aerobic digestion, thermophilic and mesophilic anaerobic digestion, based on long-term monitoring of all processes in full-scale wastewater treatment plants, with an emphasis on the efficiency in destroying pathogens. The hygienisation effect was evaluated as a removal of counts of indicator bacteria, thermotolerant coliforms and enterococci as CFU/g total sludge solids and a frequency of a positive Salmonella spp. detection. Both thermophilic technologies of municipal wastewater sludge stabilisation had the capability of producing sludge A biosolids suitable for agricultural land application when all operational parameters (mainly temperature, mixing and retention time) were stable and maintained at an appropriate level.

  3. Strain and bioprocess improvement of a thermophilic anaerobe for the production of ethanol from wood

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Herring, Christopher D.; Kenealy, William R.; Shaw, A. Joe

    Here, the thermophilic, anaerobic bacterium Thermoanaerobacterium saccharolyticum digests hemicellulose and utilizes the major sugars present in biomass. It was previously engineered to produce ethanol at yields equivalent to yeast. While saccharolytic anaerobes have been long studied as potential biomass-fermenting organisms, development efforts for commercial ethanol production have not been reported.

  4. Strain and bioprocess improvement of a thermophilic anaerobe for the production of ethanol from wood

    DOE PAGES

    Herring, Christopher D.; Kenealy, William R.; Shaw, A. Joe; ...

    2016-06-16

    Here, the thermophilic, anaerobic bacterium Thermoanaerobacterium saccharolyticum digests hemicellulose and utilizes the major sugars present in biomass. It was previously engineered to produce ethanol at yields equivalent to yeast. While saccharolytic anaerobes have been long studied as potential biomass-fermenting organisms, development efforts for commercial ethanol production have not been reported.

  5. Hydrolytic activities of extracellular enzymes in thermophilic and mesophilic anaerobic sequencing-batch reactors treating organic fractions of municipal solid wastes.

    PubMed

    Kim, Hyun-Woo; Nam, Joo-Youn; Kang, Seok-Tae; Kim, Dong-Hoon; Jung, Kyung-Won; Shin, Hang-Sik

    2012-04-01

    Extracellular enzymes offer active catalysis for hydrolysis of organic solid wastes in anaerobic digestion. To evidence the quantitative significance of hydrolytic enzyme activities for major waste components, track studies of thermophilic and mesophilic anaerobic sequencing-batch reactors (TASBR and MASBR) were conducted using a co-substrate of real organic wastes. During 1day batch cycle, TASBR showed higher amylase activity for carbohydrate (46%), protease activity for proteins (270%), and lipase activity for lipids (19%) than MASBR. In particular, the track study of protease identified that thermophilic anaerobes degraded protein polymers much more rapidly. Results revealed that differences in enzyme activities eventually affected acidogenic and methanogenic performances. It was demonstrated that the superior nature of enzymatic capability at thermophilic condition led to successive high-rate acidogenesis and 32% higher CH(4) recovery. Consequently, these results evidence that the coupling thermophilic digestion with sequencing-batch operation is a viable option to promote enzymatic hydrolysis of organic particulates. Copyright © 2012 Elsevier Ltd. All rights reserved.

  6. Enrichment of anaerobic syngas-converting bacteria from thermophilic bioreactor sludge.

    PubMed

    Alves, Joana I; Stams, Alfons J M; Plugge, Caroline M; Alves, M Madalena; Sousa, Diana Z

    2013-12-01

    Thermophilic (55 °C) anaerobic microbial communities were enriched with a synthetic syngas mixture (composed of CO, H2 , and CO2 ) or with CO alone. Cultures T-Syn and T-CO were incubated and successively transferred with syngas (16 transfers) or CO (9 transfers), respectively, with increasing CO partial pressures from 0.09 to 0.88 bar. Culture T-Syn, after 4 successive transfers with syngas, was also incubated with CO and subsequently transferred (9 transfers) with solely this substrate - cultures T-Syn-CO. Incubation with syngas and CO caused a rapid decrease in the microbial diversity of the anaerobic consortium. T-Syn and T-Syn-CO showed identical microbial composition and were dominated by Desulfotomaculum and Caloribacterium species. Incubation initiated with CO resulted in the enrichment of bacteria from the genera Thermincola and Thermoanaerobacter. Methane was detected in the first two to three transfers of T-Syn, but production ceased afterward. Acetate was the main product formed by T-Syn and T-Syn-CO. Enriched T-CO cultures showed a two-phase conversion, in which H2 was formed first and then converted to acetate. This research provides insight into how thermophilic anaerobic communities develop using syngas/CO as sole energy and carbon source can be steered for specific end products and subsequent microbial synthesis of chemicals. © 2013 Federation of European Microbiological Societies. Published by John Wiley & Sons Ltd. All rights reserved.

  7. Co-digestion of polylactide and kitchen garbage in hyperthermophilic and thermophilic continuous anaerobic process.

    PubMed

    Wang, Feng; Hidaka, Taira; Tsuno, Hiroshi; Tsubota, Jun

    2012-05-01

    Two series of two-phase anaerobic systems, consisting of a hyperthermophilic (80°C) reactor and a thermophilic (55°C) reactor, fed with a mixture of kitchen garbage (KG) and polylactide (PLA), was compared with a single-phase thermophilic reactor for the overall performance. The result indicated that ammonia addition under hyperthermophilic condition promoted the transformation of PLA particles to lactic acid. The systems with hyperthermophilic treatment had advantages on PLA transformation and methane conversion ratio to the control system. Under the organic loading rate (OLR) of 10.3 g COD/(L day), the PLA transformation ratios of the two-phase systems were 82.0% and 85.2%, respectively, higher than that of the control system (63.5%). The methane conversion ratios of the two-phase systems were 82.9% and 80.8%, respectively, higher than 70.1% of the control system. The microbial community analysis indicated that hyperthermophilic treatment is easily installed to traditional thermophilic anaerobic digestion plants without inoculation of special bacteria. Copyright © 2012 Elsevier Ltd. All rights reserved.

  8. Rapid startup of thermophilic anaerobic digester to remove tetracycline and sulfonamides resistance genes from sewage sludge.

    PubMed

    Xu, Rui; Yang, Zhao-Hui; Wang, Qing-Peng; Bai, Yang; Liu, Jian-Bo; Zheng, Yue; Zhang, Yan-Ru; Xiong, Wei-Ping; Ahmad, Kito; Fan, Chang-Zheng

    2018-01-15

    Spread of antibiotic resistance genes (ARGs) originating from sewage sludge is highlighted as an eminent health threat. This study established a thermophilic anaerobic digester using one-step startup strategy to quickly remove tetracycline and sulfonamides resistance genes from sewage sludge. At least 20days were saved in the startup period from mesophilic to thermophilic condition. Based on the results of 16S rDNA amplicons sequencing and predicted metagenomic method, the successful startup largely relied on the fast colonization of core thermophilic microbial population (e.g. Firmicutes, Proteobacteria, Actinobacteria). Microbial metabolic gene pathways for substrate degradation and methane production was also increased by one-step mode. In addition, real-time quantitative PCR approach revealed that most targeted tetracycline and sulfonamides resistance genes ARGs (sulI, tetA, tetO, tetX) were substantially removed during thermophilic digestion (removal efficiency>80%). Network analysis showed that the elimination of ARGs was attributed to the decline of their horizontal (intI1 item) and vertical (potential hosts) transfer-related elements under high-temperature. This research demonstrated that rapid startup thermophilic anaerobic digestion of wastewater solids would be a suitable technology for reducing quantities of various ARGs. Copyright © 2017 Elsevier B.V. All rights reserved.

  9. Reducing waste contamination from animal-processing plants by anaerobic thermophilic fermentation and by flesh fly digestion.

    PubMed

    Marchaim, U; Gelman, A; Braverman, Y

    2003-01-01

    There is currently no market in Israel for the large amounts of waste from fish- and poultry-processing plants. Therefore, this waste is incinerated, as part of the measures to prevent the spread of pathogens. Anaerobic methanogenic thermophilic fermentation (AMTF) of wastes from the cattle-slaughtering industry was examined previously, as an effective system to treat pathogenic bacteria, and in this article, we discuss a combined method of digestion by thermophilic anaerobic bacteria and by flesh flies, as a means of waste treatment. The AMTF process was applied to the wastes on a laboratory scale, and digestion by rearing of flesh fly (Phaenicia sericata) and housefly (Musca domestica) larvae on the untreated raw material was done on a small scale and showed remarkable weight conversion to larvae. The yield from degradation of poultry waste by flesh fly was 22.47% (SD = 3.89) and that from fish waste degradation was 35.34% (SD = 12.42), which is significantly higher than that from rearing houseflies on a regular rearing medium. Bacterial contents before and after thermophilic anaerobic digestion, as well as the changes in the chemical composition of the components during the rearing of larvae, were also examined.

  10. Occurrence and molecular characterization of cultivable mesophilic and thermophilic obligate anaerobic bacteria isolated from paper mills.

    PubMed

    Suihko, Maija-Liisa; Partanen, Laila; Mattila-Sandholm, Tiina; Raaska, Laura

    2005-08-01

    The aim of this work was to characterize the cultivable obligate anaerobic bacterial population in paper mill environments. A total of 177 anaerobically grown bacterial isolates were screened for aerotolerance, from which 67 obligate anaerobes were characterized by automated ribotyping and 41 were further identified by partial 16S rDNA sequencing. The mesophilic isolates indicated 11 different taxa (species) within the genus Clostridium and the thermophilic isolates four taxa within the genus Thermoanaerobacterium and one within Thermoanaerobacter (both formerly Clostridium). The most widespread mesophilic bacterium was closely related to C. magnum and occurred in three of four mills. One mill was contaminated with a novel mesophilic bacterium most closely related to C. thiosulfatireducens. The most common thermophile was T. thermosaccharolyticum, occurring in all four mills. The genetic relationships of the mill isolates to described species indicated that most of them are potential members of new species. On the basis of identical ribotypes clay could be identified to be the contamination source of thermophilic bacteria. Automated ribotyping can be a useful tool for the identification of clostridia as soon as comprehensive identification libraries are available.

  11. Multiple approaches to characterize the microbial community in a thermophilic anaerobic digester running on swine manure: a case study.

    PubMed

    Tuan, Nguyen Ngoc; Chang, Yi-Chia; Yu, Chang-Ping; Huang, Shir-Ly

    2014-01-01

    In this study, the first survey of microbial community in thermophilic anaerobic digester using swine manure as sole feedstock was performed by multiple approaches including denaturing gradient gel electrophoresis (DGGE), clone library and pyrosequencing techniques. The integrated analysis of 21 DGGE bands, 126 clones and 8506 pyrosequencing read sequences revealed that Clostridia from the phylum Firmicutes account for the most dominant Bacteria. In addition, our analysis also identified additional taxa that were missed by the previous researches, including members of the bacterial phyla Synergistetes, Planctomycetes, Armatimonadetes, Chloroflexi and Nitrospira which might also play a role in thermophilic anaerobic digester. Most archaeal 16S rRNA sequences could be assigned to the order Methanobacteriales instead of Methanomicrobiales comparing to previous studies. In addition, this study reported that the member of Methanothermobacter genus was firstly found in thermophilic anaerobic digester. Copyright © 2014 Elsevier GmbH. All rights reserved.

  12. Inactivation of Clostridium difficile in sewage sludge by anaerobic thermophilic digestion.

    PubMed

    Xu, Changyun; Salsali, Hamidreza; Weese, Scott; Warriner, Keith

    2016-01-01

    There has been an increase in community-associated Clostridium difficile infections with biosolids derived from wastewater treatment being identified as one potential source. The current study evaluated the efficacy of thermophilic digestion in decreasing levels of C. difficile ribotype 078 associated with sewage sludge. Five isolates of C. difficile 078 were introduced (final density of 5 log CFU/g) into digested sludge and subjected to anaerobic digestion at mesophilic (36 or 42 °C) or thermophilic (55 °C) temperatures for up to 60 days. It was found that mesophilic digestion at 36 °C did not result in a significant reduction in C. difficile spore levels. In contrast, thermophilic sludge digestion reduced endospore levels at a rate of 0.19-2.68 log CFU/day, depending on the strain tested. The mechanism of lethality was indirect - by stimulating germination then inactivating the resultant vegetative cells. Acidification of sludge by adding acetic acid (6 g/L) inhibited the germination of spores regardless of the sludge digestion temperature. In conclusion, thermophilic digestion can be applied to reduce C. difficile in biosolids, thereby reducing the environmental burden of the enteric pathogen.

  13. Mesophilic and thermophilic anaerobic biodegradability of water hyacinth pre-treated at 80 degrees C.

    PubMed

    Ferrer, Ivet; Palatsi, Jordi; Campos, Elena; Flotats, Xavier

    2010-10-01

    Water hyacinth (Eichornia crassipes) is a fast growing aquatic plant which causes environmental problems in continental water bodies. Harvesting and handling this plant becomes an issue, and focus has been put on the research of treatment alternatives. Amongst others, energy production through biomethanation has been proposed. The aim of this study was to assess the anaerobic biodegradability of water hyacinth under mesophilic and thermophilic conditions. The effect of a thermal sludge pre-treatment at 80 degrees C was also evaluated. To this end, anaerobic biodegradability tests were carried out at 35 degrees C and 55 degrees C, with raw and pre-treated water hyacinth. According to the results, the thermal pre-treatment enhanced the solubilisation of water hyacinth (i.e. increase in the soluble to total chemical oxygen demand (COD)) from 4% to 12% after 30 min. However, no significant effect was observed on the methane yields (150-190 L CH(4)/kg volatile solids). Initial methane production rates for thermophilic treatments were two fold those of mesophilic ones (6-6.5L vs. 3-3.5 L CH(4)/kg COD x day). Thus, higher methane production rates might be expected from thermophilic reactors working at short retention times. The study of longer low temperature pre-treatments or pre-treatments at elevated temperatures coupled to thermophilic reactors should be considered in the future. (c) 2009 Elsevier Ltd. All rights reserved.

  14. Treatment of kraft evaporator condensate using a thermophilic submerged anaerobic membrane bioreactor.

    PubMed

    Liao, B Q; Xie, K; Lin, H J; Bertoldo, Daniel

    2010-01-01

    The feasibility of using a thermophilic submerged anaerobic membrane bioreactor (SAnMBR) for kraft evaporator condensate treatment was studied at 55+/-1 degrees C over 6.5 months. Under tested organic loading rate of 1-7 kg COD/m(3) day, a soluble COD removal efficiency of 85-97% was obtained. The methane production rate was 0.35+/-0.1 L methane/g COD and the produced biogas was of excellent fuel quality with 80-90% methane. A higher membrane fouling rate was related to the presence of a larger portion of fine colloidal particles (1-10 mum). The thermophilic SAnMBR was sensitive to the presence of toxic compounds in feed and unexpected pH probe failure (leading to a higher pH). Feed toxic shock caused sludge deflocculation and thus deteriorated membrane performance. Operating the reactor as a conventional anaerobic reactor to waste some of the fine flocs in treated effluent during the start-up process was an effective strategy to reduce membrane fouling. The experimental results from this study indicate that treatment of kraft evaporator condensate is feasible in terms of COD removal and biogas production using thermophilic SAnMBRs but pre-treatment may be needed to remove toxic sulfur compounds and membrane fouling caused by the large portion of fine particles may be a challenge.

  15. Kinetic characterization of thermophilic and mesophilic anaerobic digestion for coffee grounds and waste activated sludge.

    PubMed

    Li, Qian; Qiao, Wei; Wang, Xiaochang; Takayanagi, Kazuyuki; Shofie, Mohammad; Li, Yu-You

    2015-02-01

    This study was conducted to characterize the kinetics of an anaerobic process (hydrolysis, acetogenesis, acidogenesis and methanogenesis) under thermophilic (55 °C) and mesophilic (35 °C) conditions with coffee grounds and waste activated sludge (WAS) as the substrates. Special focus was given to the kinetics of propionic acid degradation to elucidate the accumulation of VFAs. Under the thermophilic condition, the methane production rate of all substrates (WAS, ground coffee and raw coffee) was about 1.5 times higher than that under the mesophilic condition. However, the effects on methane production of each substrate under the thermophilic condition differed: WAS increased by 35.8-48.2%, raw coffee decreased by 76.3-64.5% and ground coffee decreased by 74.0-57.9%. Based on the maximum reaction rate (Rmax) of each anaerobic stage obtained from the modified Gompertz model, acetogenesis was found to be the rate-limiting step for coffee grounds and WAS. This can be explained by the kinetics of propionate degradation under thermophilic condition in which a long lag-phase (more than 18 days) was observed, although the propionate concentration was only 500 mg/L. Under the mesophilic condition, acidogenesis and hydrolysis were found to be the rate-limiting step for coffee grounds and WAS, respectively. Even though reducing the particle size accelerated the methane production rate of coffee grounds, but did not change the rate-limiting step: acetogenesis in thermophilic and acidogenesis in mesophilic. Copyright © 2014 Elsevier Ltd. All rights reserved.

  16. Start-up, steady state performance and kinetic evaluation of a thermophilic integrated anaerobic-aerobic bioreactor (IAAB).

    PubMed

    Chan, Yi Jing; Chong, Mei Fong; Law, Chung Lim

    2012-12-01

    Thermophilic treatment of palm oil mill effluent (POME) was studied in a novel integrated anaerobic-aerobic bioreactor (IAAB). The IAAB was subjected to a program of steady-state operation over a range of organic loading rate (OLR)s, up to 30 g COD/L day in order to evaluate its treatment capacity. The thermophilic IAAB achieved high chemical oxygen demand (COD), biochemical oxygen demand (BOD) and total suspended solids (TSS) removal efficiencies of more than 99% for OLR up to 18.5 g COD/L day. High methane yield of 0.32 LCH(4) (STP)/g COD(removed) with compliance of the final treated effluent to the discharge limit were achieved. This is higher than that of the mesophilic system due to the higher maximum specific growth rate (μ(max)) of the thermophilic microorganisms. Besides, coupling the model of Grau second order model (anaerobic system) with the model of Monod (aerobic system) will completely define the IAAB system. Copyright © 2012 Elsevier Ltd. All rights reserved.

  17. Thermophilic anaerobic digestion in compact systems: investigations by modern microbiological techniques and mathematical simulation.

    PubMed

    Lübken, M; Wichern, M; Letsiou, I; Kehl, O; Bischof, F; Horn, H

    2007-01-01

    Thermophilic anaerobic digestion in compact systems can be an economical and ecological reasonable decentralised process technique, especially for rural areas. Thermophilic process conditions are important for a sufficient removal of pathogens. The high energy demand, however, can make such systems unfavourable in terms of energy costs. This is the case when low concentrated wastewater is treated or the system is operated at low ambient temperatures. In this paper we present experimental results of a compact thermophilic anaerobic system obtained with fluorescent in situ hybridisation (FISH) analysis and mathematical simulation. The system was operated with faecal sludge for a period of 135 days and with a model substrate consisting of forage and cellulose for a period of 60 days. The change in the microbial community due to the two different substrates treated could be well observed by the FISH analysis. The Anaerobic Digestion Model no. 1 (ADM1) was used to evaluate system performance at different temperature conditions. The model was extended to contribute to decreased methanogenic activity at lower temperatures and was used to calculate energy production. A model was developed to calculate the major parts of energy consumed by the digester itself at different temperature conditions. It was demonstrated by the simulation study that a reduction of the process temperature can lead to higher net energy yield. The simulation study additionally showed that the effect of temperature on the energy yield is higher when a substrate is treated with high protein content.

  18. Inferring microbial interactions in thermophilic and mesophilic anaerobic digestion of hog waste.

    PubMed

    Shaw, Grace Tzun-Wen; Liu, An-Chi; Weng, Chieh-Yin; Chou, Chu-Yang; Wang, Daryi

    2017-01-01

    Anaerobic digestion (AnD) is a microbiological process that converts organic waste materials into biogas. Because of its high methane content, biogas is a combustible energy source and serves as an important environmental technology commonly used in the management of animal waste generated on large animal farms. Much work has been done on hardware design and process engineering for the generation of biogas. However, little is known about the complexity of the microbiology in this process. In particular, how microbes interact in the digester and eventually breakdown and convert organic matter into biogas is still regarded as a "black box." We used 16S rRNA sequencing as a tool to study the microbial community in laboratory hog waste digesters under tightly controlled conditions, and systematically unraveled the distinct interaction networks of two microbial communities from mesophilic (MAnD) and thermophilic anaerobic digestion (TAnD). Under thermophilic conditions, the well-known association between hydrogen-producing bacteria, e.g., Ruminococcaceae and Prevotellaceae, and hydrotrophic methanogens, Methanomicrobiaceae, was reverse engineered by their interactive topological niches. The inferred interaction network provides a sketch enabling the determination of microbial interactive relationships that conventional strategy of finding differential taxa was hard to achieve. This research is still in its infancy, but it can help to depict the dynamics of microbial ecosystems and to lay the groundwork for understanding how microorganisms cohabit in the anaerobic digester.

  19. High performance biological methanation in a thermophilic anaerobic trickle bed reactor.

    PubMed

    Strübing, Dietmar; Huber, Bettina; Lebuhn, Michael; Drewes, Jörg E; Koch, Konrad

    2017-12-01

    In order to enhance energy efficiency of biological methanation of CO 2 and H 2 , this study investigated the performance of a thermophilic (55°C) anaerobic trickle bed reactor (ATBR) (58.1L) at ambient pressure. With a methane production rate of up to 15.4m 3 CH4 /(m 3 trickle bed ·d) at methane concentrations above 98%, the ATBR can easily compete with the performance of other mixed culture methanation reactors. Control of pH and nutrient supply turned out to be crucial for stable operation and was affected significantly by dilution due to metabolic water production, especially during demand-orientated operation. Considering practical applications, inoculation with digested sludge, containing a diverse biocenosis, showed high adaptive capacity due to intrinsic biological diversity. However, no macroscopic biofilm formation was observed at thermophilic conditions even after 313days of operation. The applied approach illustrates the high potential of thermophilic ATBRs as a very efficient energy conversion and storage technology. Copyright © 2017 Elsevier Ltd. All rights reserved.

  20. Dry co-digestion of sewage sludge and rice straw under mesophilic and thermophilic anaerobic conditions.

    PubMed

    Chu, Xiangqian; Wu, Guangxue; Wang, Jiaquan; Hu, Zhen-Hu

    2015-12-01

    Dry anaerobic digestion of sewage sludge can recover biogas as energy; however, its low C/N ratio limits it as a single substrate in the anaerobic digestion. Rice straw is an abundant agricultural residue in China, which is rich in carbon and can be used as carbon source. In the present study, the performance of dry co-digestion of sewage sludge and rice straw was investigated under mesophilic (35 °C) and thermophilic (55 °C) conditions. The operational factors impacting dry co-digestion of sewage sludge and rice straw such as C/N ratio, moisture content, and initial pH were explored under mesophilic conditions. The results show that low C/N ratios resulted in a higher biogas production rate, but a lower specific biogas yield; low moisture content of 65 % resulted in the instability of the digestion system and a low specific biogas yield. Initial pH ranging 7.0-9.0 did not affect the performance of the anaerobic digestion. The C/N ratio of 26-29:1, moisture content of 70-80 %, and pH 7.0-9.0 resulted in good performance in the dry mesophilic co-digestion of sewage sludge and rice straw. As compared with mesophilic digestion, thermophilic co-digestion of sewage sludge and rice straw significantly enhanced the degradation efficiency of the substrates and the specific biogas yield (p < 0.05) at the conditions of C/N ratio 26:1, moisture content 80 %, and natural initial pH. Although high concentrations of ammonia-nitrogen (NH4-N, 1500 mg/kg wet weight) were formed during thermophilic digestion, there was no obvious inhibition occurred. The results indicated that rice straw can be used as carbon source for the dry co-digestion of sewage sludge under mesophilic and thermophilic conditions.

  1. Entrapment of anaerobic thermophilic and hyperthermophilic marine micro-organisms in a gellan/xanthan matrix.

    PubMed

    Landreau, M; Duthoit, F; Claeys-Bruno, M; Vandenabeele-Trambouze, O; Aubry, T; Godfroy, A; Le Blay, G

    2016-06-01

    The aims of this study were (i) to develop a protocol for the entrapment of anaerobic (hyper)thermophilic marine micro-organisms; (ii) to test the use of the chosen polymers in a range of physical and chemical conditions and (iii) to validate the method with batch cultures. The best conditions for immobilization were obtained at 80°C with gellan and xanthan gums. After 5-week incubation, beads showed a good resistance to all tested conditions except those simultaneously including high temperature (100°C), low NaCl (<0∙5 mol l(-1) ) and extreme pH (4/8). To confirm the method efficiency, batch cultures with immobilized Thermosipho sp. strain AT1272 and Thermococcus kodakarensis strain KOD1 showed an absence of detrimental effect on cell viability and a good growth within and outside the beads. This suggests that entrapment in a gellan-xanthan matrix could be employed for the culture of anaerobic (hyper)thermophilic marine micro-organisms. (Hyper)thermophilic marine micro-organisms possess a high biotechnological potential. Generally microbial cells are grown as free-cell cultures. The use of immobilized cells may offer several advantages such as protection against phage attack, high cell biomass and better production rate of desired metabolites. © 2016 The Society for Applied Microbiology.

  2. Start-up of thermophilic-dry anaerobic digestion of OFMSW using adapted modified SEBAC inoculum.

    PubMed

    Fdéz-Güelfo, L A; Alvarez-Gallego, C; Sales Márquez, D; Romero García, L I

    2010-12-01

    The work presented here concerns the start-up and stabilization stages of a Continuous Stirred Tank Reactor (CSTR) semicontinuously fed for the treatment of the Organic Fraction of Municipal Solid Waste (OFMSW) through anaerobic digestion at thermophilic temperature range (55 degrees C) and dry conditions (30% Total Solids). The procedure reported involves two novel aspects with respect to other start-up and stabilization protocols reported in the literature. The novel aspects concern the adaptation of the inoculum to both the operating conditions (thermophilic and dry) and to the type of waste by employing a modified SEBAC (Sequential Batch Anaerobic Composting) system and, secondly, the direct start-up of the process in a thermophilic temperature regime and feeding of the system from the first day of operation. In this way a significant reduction in the start-up time and stabilization is achieved i.e. 110 days in comparison to 250 days for the processes reported by other authors for the same type of waste and digester. The system presents suitable operational conditions to stabilize the reactor at SRT of 35 days, with a maximum biogas production of 1.944 LR/L.d with a CH(4) and CO(2) percentage of 25.27% and 68.15%, respectively. 2010 Elsevier Ltd. All rights reserved.

  3. Inferring microbial interactions in thermophilic and mesophilic anaerobic digestion of hog waste

    PubMed Central

    Shaw, Grace Tzun-Wen; Liu, An-Chi; Weng, Chieh-Yin; Chou, Chu-Yang

    2017-01-01

    Anaerobic digestion (AnD) is a microbiological process that converts organic waste materials into biogas. Because of its high methane content, biogas is a combustible energy source and serves as an important environmental technology commonly used in the management of animal waste generated on large animal farms. Much work has been done on hardware design and process engineering for the generation of biogas. However, little is known about the complexity of the microbiology in this process. In particular, how microbes interact in the digester and eventually breakdown and convert organic matter into biogas is still regarded as a “black box.” We used 16S rRNA sequencing as a tool to study the microbial community in laboratory hog waste digesters under tightly controlled conditions, and systematically unraveled the distinct interaction networks of two microbial communities from mesophilic (MAnD) and thermophilic anaerobic digestion (TAnD). Under thermophilic conditions, the well-known association between hydrogen-producing bacteria, e.g., Ruminococcaceae and Prevotellaceae, and hydrotrophic methanogens, Methanomicrobiaceae, was reverse engineered by their interactive topological niches. The inferred interaction network provides a sketch enabling the determination of microbial interactive relationships that conventional strategy of finding differential taxa was hard to achieve. This research is still in its infancy, but it can help to depict the dynamics of microbial ecosystems and to lay the groundwork for understanding how microorganisms cohabit in the anaerobic digester. PMID:28732056

  4. Mesophilic and thermophilic anaerobic biodegradability of water hyacinth pre-treated at 80 {sup o}C

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Ferrer, Ivet, E-mail: ivet.ferrer@upc.ed; Palatsi, Jordi; Campos, Elena

    2010-10-15

    Water hyacinth (Eichornia crassipes) is a fast growing aquatic plant which causes environmental problems in continental water bodies. Harvesting and handling this plant becomes an issue, and focus has been put on the research of treatment alternatives. Amongst others, energy production through biomethanation has been proposed. The aim of this study was to assess the anaerobic biodegradability of water hyacinth under mesophilic and thermophilic conditions. The effect of a thermal sludge pre-treatment at 80 {sup o}C was also evaluated. To this end, anaerobic biodegradability tests were carried out at 35 {sup o}C and 55 {sup o}C, with raw and pre-treatedmore » water hyacinth. According to the results, the thermal pre-treatment enhanced the solubilisation of water hyacinth (i.e. increase in the soluble to total chemical oxygen demand (COD)) from 4% to 12% after 30 min. However, no significant effect was observed on the methane yields (150-190 L CH{sub 4}/kg volatile solids). Initial methane production rates for thermophilic treatments were two fold those of mesophilic ones (6-6.5 L vs. 3-3.5 L CH{sub 4}/kg COD.day). Thus, higher methane production rates might be expected from thermophilic reactors working at short retention times. The study of longer low temperature pre-treatments or pre-treatments at elevated temperatures coupled to thermophilic reactors should be considered in the future.« less

  5. Recurrence of fecal coliforms and Salmonella species in biosolids following thermophilic anaerobic digestion.

    PubMed

    Iranpour, Reza; Cox, Huub H J

    2006-09-01

    The U.S. Environmental Protection Agency (U.S. EPA) Part 503 Biosolids Rule requires the fecal coliform (indicator) or Salmonella species (pathogen) density requirements for Class A biosolids to be met at the last point of plant control (truck-loading facility and/or farm for land application). The three Southern Californian wastewater treatment plants in this study produced biosolids by thermophilic anaerobic digestion and all met the Class A limits for both fecal coliforms and Salmonella sp. in the digester outflow biosolids. At two plants, however, a recurrence of fecal coliforms was observed in postdigestion biosolids, which caused exceedance of the Class A limit for fecal coliforms at the truck-loading facility and farm for land application. Comparison of observations at the three plants and further laboratory tests indicated that the recurrence of fecal coliforms can possibly be related to the following combination of factors: (1) incomplete destruction of fecal coliforms during thermophilic anaerobic digestion, (2) contamination of Class A biosolids with fecal coliforms from external sources during postdigestion, (3) a large drop of the postdigestion biosolids temperature to below the maximum for fecal coliform growth, (4) an unknown effect of biosolids dewatering in centrifuges. At Hyperion Treatment Plant (City of Los Angeles, California), fecal coliform recurrence could be prevented by the following: (1) complete conversion to thermophilic operation to exclude contamination by mesophilically digested biosolids and (2) insulation and electrical heat-tracing of postdigestion train for maintaining a high biosolids temperature in postdigestion.

  6. Kinetics of thermophilic anaerobes in fixed-bed reactors.

    PubMed

    Perez, M; Romero, L I; Sales, D

    2001-08-01

    The main objective of this study is to estimate growth kinetic constants and the concentration of "active" attached biomass in two anaerobic thermophilic reactors which contain different initial sizes of immobilized anaerobic mixed cultures and decompose distillery wastewater. This paper studies the substrate decomposition in two lab-scale fixed-bed reactors operating at batch conditions with corrugated tubes as support media. It can be demonstrated that high micro-organisms-substrate ratios favor the degradation activity of the different anaerobic cultures, allowing the stable operation without lag-phases and giving better quality in effluent. The kinetic parameters obtained--maximum specific growth rates (mu(max)), non-biodegradable substrate (S(NB)) and "active or viable biomass" concentrations (X(V0))--were obtained by applying the Romero kinetic model [L.I. Romero, 1991. Desarrollo de un modelo matemático general para los procesos fermentativos, Cinética de la degradación anaerobia, Ph.D. Thesis, University of Cádiz (Spain), Serv. Pub. Univ. Cádiz], with COD as substrate and methane (CH4) as the main product of the anaerobic process. This method is suitable to calculate and to differentiate the main kinetic parameters of both the total anaerobic mixed culture and the methanogenic population. Comparison of experimental measured concentration of volatile attached solids (VS(att)) in both reactors with the estimated "active" biomass concentrations obtained by applying Romero kinetic model [L.I. Romero, 1991. Desarrollo de un modelo matemático general para los procesos fermentativos, Cinética de la degradación anaerobia, Ph.D. Thesis, University of Cádiz (Spain), Serv. Pub. Univ. Cádiz] shows that a large amount of inert matter is present in the fixed-bed reactor.

  7. A comparative study of thermophilic and mesophilic anaerobic co-digestion of food waste and wheat straw: Process stability and microbial community structure shifts.

    PubMed

    Shi, Xuchuan; Guo, Xianglin; Zuo, Jiane; Wang, Yajiao; Zhang, Mengyu

    2018-05-01

    Renewable energy recovery from organic solid waste via anaerobic digestion is a promising way to provide sustainable energy supply and eliminate environmental pollution. However, poor efficiency and operational problems hinder its wide application of anaerobic digestion. The effects of two key parameters, i.e. temperature and substrate characteristics on process stability and microbial community structure were studied using two lab-scale anaerobic reactors under thermophilic and mesophilic conditions. Both the reactors were fed with food waste (FW) and wheat straw (WS). The organic loading rates (OLRs) were maintained at a constant level of 3 kg VS/(m 3 ·d). Five different FW:WS substrate ratios were utilized in different operational phases. The synergetic effects of co-digestion improved the stability and performance of the reactors. When FW was mono-digested, both reactors were unstable. The mesophilic reactor eventually failed due to volatile fatty acid accumulation. The thermophilic reactor had better performance compared to mesophilic one. The biogas production rate of the thermophilic reactor was 4.9-14.8% higher than that of mesophilic reactor throughout the experiment. The shifts in microbial community structures throughout the experiment in both thermophilic and mesophilic reactors were investigated. With increasing FW proportions, bacteria belonging to the phylum Thermotogae became predominant in the thermophilic reactor, while the phylum Bacteroidetes was predominant in the mesophilic reactor. The genus Methanosarcina was the predominant methanogen in the thermophilic reactor, while the genus Methanothrix remained predominant in the mesophilic reactor. The methanogenesis pathway shifted from acetoclastic to hydrogenotrophic when the mesophilic reactor experienced perturbations. Moreover, the population of lignocellulose-degrading microorganisms in the thermophilic reactor was higher than those in mesophilic reactor, which explained the better

  8. Cassava Stillage Treatment by Thermophilic Anaerobic Continuously Stirred Tank Reactor (CSTR)

    NASA Astrophysics Data System (ADS)

    Luo, Gang; Xie, Li; Zou, Zhonghai; Zhou, Qi

    2010-11-01

    This paper assesses the performance of a thermophilic anaerobic Continuously Stirred Tank Reactor (CSTR) in the treatment of cassava stillage under various organic loading rates (OLRs) without suspended solids (SS) separation. The reactor was seeded with mesophilic anaerobic granular sludge, and the OLR increased by increments to 13.80 kg COD/m3/d (HRT 5d) over 80 days. Total COD removal efficiency remained stable at 90%, with biogas production at 18 L/d (60% methane). Increase in the OLR to 19.30 kg COD/m3/d (HRT 3d), however, led to a decrease in TCOD removal efficiency to 79% due to accumulation of suspended solids and incomplete degradation after shortened retention time. Reactor performance subsequently increased after OLR reduction. Alkalinity, VFA and pH levels were not significantly affected by OLR variation, indicating that no additional alkaline or pH adjustment is required. More than half of the SS in the cassava stillage could be digested in the process when HRT was 5 days, which demonstrated the suitability of anaerobic treatment of cassava stillage without SS separation.

  9. Description of a new anaerobic thermophilic bacterium, Thermoanaerobacterium butyriciformans sp. nov.

    PubMed

    López, G; Cañas-Duarte, S J; Pinzón-Velasco, A M; Vega-Vela, N E; Rodríguez, M; Restrepo, S; Baena, S

    2017-03-01

    Strain USBA-019 T , an anaerobic and thermophilic strain, was identified as a new member of the genus Thermoanaerobacterium. USBA-019 T cells are gram-positive, strictly anaerobic, thermophilic, chemoorganotrophic, moderately acidophilic, non-motile, endospore-forming, slightly curved, and rod-shaped. Cells measure 0.4×3.0-7.0μm. Optimal growth occurs at 50-55°C (35-65°C). Optimum pH is 5.0-5.5 (4.0-8.5). Thiosulfate, elemental sulfur and nitrate were utilized as electron acceptors. Fermentation of glucose, lactose, cellobiose, galactose, arabinose, xylose, starch and xylan primarily produced acetate and butyrate. Xylan, starch and cellobiose produced ethanol and starch, cellobiose, galactose, arabinose and mannose produced lactic acid. Phylogenetic analyses based on 16S rRNA gene sequence comparison and genomic relatedness indices show the close relation of USBA-019 T to Thermoanaerobacterium thermostercoris and Thermoanaerobacterium aotearoense (similarity value: 99%). Hybridization of USBA-019 T , Th. thermostercoris DSM22141 T and Th. aotearoense DMS10170 T found DNA-DNA relatedness of 33.2% and 18.2%, respectively. Based on phenotypic, chemotaxonomic and phylogenetic evidence, along with low identity at whole genome level, USBA-019 T is a novel species of the genus Thermoanaerobacterium which we propose to name Thermoanaerobacterium butyriciformans sp. nov. The type strain is USBA-019 T (=CMPUJ U-019 T =DSM 101588 T ). Copyright © 2016 Elsevier GmbH. All rights reserved.

  10. Effects of total solids content on waste activated sludge thermophilic anaerobic digestion and its sludge dewaterability.

    PubMed

    Wang, Tianfeng; Chen, Jie; Shen, Honglang; An, Dong

    2016-10-01

    The role of total solids content on sludge thermophilic anaerobic digestion was investigated in batch reactors. A range of total solids content from 2% to 10% was evaluated with two replicates. The lowest inhibitory concentration for free ammonia and total ammonia of sludge thermophilic anaerobic digestion was 110.9-171.4mg/L and 1313.1-1806.7mg/L, respectively. The volumetric biogas production rate increased with increasing of total solids content, but the corresponding biogas yield per gram volatile solid decreased. The result of normalized capillary suction time indicated that the dewaterability of digested sludge at high total solids content was poor, while solid content of sediment obtained by centrifuging sludge at 2000g for 10min increased with increasing of total solids content of sludge. The results suggest that thickened sludge mixed with dewatered sludge at an appropriate ratio could get high organic loading rate, high biogas yield and adequate dewatering effort. Copyright © 2016 Elsevier Ltd. All rights reserved.

  11. Mesophilic versus thermophilic anaerobic digestion of cattle manure: methane productivity and microbial ecology

    PubMed Central

    Moset, Veronica; Poulsen, Morten; Wahid, Radziah; Højberg, Ole; Møller, Henrik Bjarne

    2015-01-01

    In this study, productivity and physicochemical and microbiological (454 sequencing) parameters, as well as environmental criteria, were investigated in anaerobic reactors to contribute to the ongoing debate about the optimal temperature range for treating animal manure, and expand the general knowledge on the relation between microbiological and physicochemical process indicators. For this purpose, two reactor sizes were used (10 m3 and 16 l), in which two temperature conditions (35°C and 50°C) were tested. In addition, the effect of the hydraulic retention time was evaluated (16 versus 20 days). Thermophilic anaerobic digestion showed higher organic matter degradation (especially fiber), higher pH and higher methane (CH4) yield, as well as better percentage of ultimate CH4 yield retrieved and lower residual CH4 emission, when compared with mesophilic conditions. In addition, lower microbial diversity was found in the thermophilic reactors, especially for Bacteria, where a clear intensification towards Clostridia class members was evident. Independent of temperature, some similarities were found in digestates when comparing with animal manure, including low volatile fatty acids concentrations and a high fraction of Euryarchaeota in the total microbial community, in which members of Methanosarcinales dominated for both temperature conditions; these indicators could be considered a sign of process stability. PMID:25737010

  12. Thermophilic anaerobic co-digestion of cattle manure with agro-wastes and energy crops: comparison of pilot and full scale experiences.

    PubMed

    Cavinato, C; Fatone, F; Bolzonella, D; Pavan, P

    2010-01-01

    The paper deals with the benefits coming from the application of a proper process temperature (55 degrees C) instead of a 'reduced' thermophilic range (47 degrees C), that is often applied in European anaerobic co-digestion plants. The experimental work has pointed out that biogas production improve from 0.45 to 0.62 m(3)/kg VS operating at proper thermophilic conditions. Moreover, also methane content was higher: from 52% to 61%. A general improvement in digester behaviour was clear also considering the stability parameters comparison (pH, ammonia, VFA content). The second part of the study takes into account the economic aspects related to the capital cost of anaerobic digestion treatment with a 1 MW co-generation unit fro heat and power production (CHP). Moreover, the economic balance was also carried out considering the anaerobic supernatants treatment for nitrogen removal. The simulation showed how a pay-back-time of 2.5 yr and between 3 and 5 yr respectively could be determined when the two options of anaerobic digestion only and together with the application of a nitrogen removal process were considered.

  13. Ultrasound-Mediated DNA Transformation in Thermophilic Gram-Positive Anaerobes

    PubMed Central

    Ji, Yuetong; He, Zhili; Pu, Yunting; Zhou, Jizhong; Xu, Jian

    2010-01-01

    Background Thermophilic, Gram-positive, anaerobic bacteria (TGPAs) are generally recalcitrant to chemical and electrotransformation due to their special cell-wall structure and the low intrinsic permeability of plasma membranes. Methodology/Principal Findings Here we established for any Gram-positive or thermophiles an ultrasound-based sonoporation as a simple, rapid, and minimally invasive method to genetically transform TGPAs. We showed that by applying a 40 kHz ultrasound frequency over a 20-second exposure, Texas red-conjugated dextran was delivered with 27% efficiency into Thermoanaerobacter sp. X514, a TGPA that can utilize both pentose and hexose for ethanol production. Experiments that delivered plasmids showed that host-cell viability and plasmid DNA integrity were not compromised. Via sonoporation, shuttle vectors pHL015 harboring a jellyfish gfp gene and pIKM2 encoding a Clostridium thermocellum β-1,4-glucanase gene were delivered into X514 with an efficiency of 6×102 transformants/µg of methylated DNA. Delivery into X514 cells was confirmed via detecting the kanamycin-resistance gene for pIKM2, while confirmation of pHL015 was detected by visualization of fluorescence signals of secondary host-cells following a plasmid-rescue experiment. Furthermore, the foreign β-1,4-glucanase gene was functionally expressed in X514, converting the host into a prototypic thermophilic consolidated bioprocessing organism that is not only ethanologenic but cellulolytic. Conclusions/Significance In this study, we developed an ultrasound-based sonoporation method in TGPAs. This new DNA-delivery method could significantly improve the throughput in developing genetic systems for TGPAs, many of which are of industrial interest yet remain difficult to manipulate genetically. PMID:20838444

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

    PubMed

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

    2011-01-01

    The methanogenic pathway and microbial community in a thermophilic anaerobic digestion process of organic solid waste were investigated in a continuous-flow stirred-tank reactor using artificial garbage slurry as a feedstock. The decomposition pathway of acetate, a significant precursor of CH(4) and a key intermediate metabolite in the anaerobic digestion process, was analyzed by using stable isotopes. A tracer experiment using (13)C-labeled acetate revealed that approximately 80% of the acetate was decomposed via a non-aceticlastic oxidative pathway, whereas the remainder was converted to methane via an aceticlastic pathway. Archaeal 16S rRNA analyses demonstrated that the hydrogenotrophic methanogens Methanoculleus spp. accounted for >90% of detected methanogens, and the aceticlastic methanogens Methanosarcina spp. were the minor constituents. The clone library targeting bacterial 16S rRNA indicated the predominance of the novel Thermotogales bacterium (relative abundance: ~53%), which is related to anaerobic acetate oxidizer Thermotoga lettingae TMO, although the sequence similarity was low. Uncultured bacteria that phylogenetically belong to municipal solid waste cluster I were also predominant in the microflora (~30%). These results imply that the microbial community in the thermophilic degrading process of organic solid waste consists exclusively of unidentified bacteria, which efficiently remove acetate through a non-aceticlastic oxidative pathway. Copyright © 2010 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.

  15. Community analysis of hydrogen-producing extreme thermophilic anaerobic microflora enriched from cow manure with five substrates.

    PubMed

    Yokoyama, Hiroshi; Moriya, Naoko; Ohmori, Hideyuki; Waki, Miyoko; Ogino, Akifumi; Tanaka, Yasuo

    2007-11-01

    The present study analyzed the community structures of anaerobic microflora producing hydrogen under extreme thermophilic conditions by two culture-independent methods: denaturing gradient gel electrophoresis (DGGE) and clone library analyses. Extreme thermophilic microflora (ETM) was enriched from cow manure by repeated batch cultures at 75 degrees C, using a substrate of xylose, glucose, lactose, cellobiose, or soluble starch, and produced hydrogen at yields of 0.56, 2.65, 2.17, 2.68, and 1.73 mol/mol-monosaccharide degraded, respectively. The results from the DGGE and clone library analyses were consistent and demonstrated that the community structures of ETM enriched with the four hexose-based substrates (glucose, lactose, cellobiose, and soluble starch) consisted of a single species, closely related to a hydrogen-producing extreme thermophile, Caldoanaerobacter subterraneus, with diversity at subspecies levels. The ETM enriched with xylose was more diverse than those enriched with the other substrates, and contained the bacterium related to C. subterraneus and an unclassified bacterium, distantly related to a xylan-degrading and hydrogen-producing extreme thermophile, Caloramator fervidus.

  16. Thermophilic two-stage dry anaerobic digestion of model garbage with ammonia stripping.

    PubMed

    Yabu, Hironori; Sakai, Chikako; Fujiwara, Tomoko; Nishio, Naomichi; Nakashimada, Yutaka

    2011-03-01

    To avoid the inhibition of methane production by ammonia that occurs during the degradation of garbage, anaerobic digestion with prior ammonia production and subsequent stripping was investigated. In the ammonia production phase, the maximum ammonia concentration was approximately 2800 mg N/kg of total wet sludge in the range of 4 days of sludge retention time, indicating that only 43% of total nitrogen in the model garbage was converted to ammonia. The model garbage from which ammonia was produced and stripped was subjected to semi-continuous thermophilic dry anaerobic digestion over 180 days. The gas yield was in the range of 0.68 to 0.75 Nm(3)/kg volatile solid, and it decreased with the decrease of the sludge retention time. The ammonia-nitrogen concentration in the sludge was kept below 3000 mg N/kg total wet sludge. Microbial community structure analysis revealed that the phylum Firmicutes dominated in the ammonia production, but the community structure changed at different sludge retention times. In dry anaerobic digestion, the dominant bacteria shifted from the phylum Thermotogae to Firmicutes. The dominant archaeon was the genus Methanothermobacter, but the ratio of Methanosarcina increased during the process of dry anaerobic digestion. Copyright © 2010 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.

  17. An explanation of the methanogenic pathway for methane production in anaerobic digestion of nitrogen-rich materials under mesophilic and thermophilic conditions.

    PubMed

    Yin, Dong-Min; Westerholm, Maria; Qiao, Wei; Bi, Shao-Jie; Wandera, Simon M; Fan, Run; Jiang, Meng-Meng; Dong, Ren-Jie

    2018-05-18

    The impact of temperature on the anaerobic digestion of chicken manure was investigated by studying the process performance and pathway for continuously-fed digesters under mesophilic and thermophilic conditions. The mesophilic digester obtained a 15% higher methane yield compared with the thermophilic digester. Mesophilic and thermophilic digester had free ammonia of 31 and 145 mg/L, respectively. The stable carbon isotope analysis indicated that 41% and 50% of acetate was converted to methane through the syntrophic acetate oxidation and hydrogenotrophic methanogenesis (SAO-HM) pathway under mesophilic and thermophilic conditions, respectively. The genus Pseudomonas represented 10% and 16% under mesophilic and thermophilic conditions, respectively. A high abundance of the methanogens genus Methanoculleus (94% of total methanogens) in mesophilic and the genus Methanothermobacter (96%) in thermophilic digesters indicated they were the main hydrogenotrophic partners in SAO. The present study therefore illustrated that methanogenic pathway shifting, induced by free ammonia, closely correlated to the process performance. Copyright © 2018. Published by Elsevier Ltd.

  18. High rate mesophilic, thermophilic, and temperature phased anaerobic digestion of waste activated sludge: A pilot scale study

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Bolzonella, David, E-mail: david.bolzonella@univr.it; Cavinato, Cristina, E-mail: cavinato@unive.it; Fatone, Francesco, E-mail: francesco.fatone@univr.it

    2012-06-15

    Highlights: Black-Right-Pointing-Pointer High temperatures were tested in single and two-stage anaerobic digestion of waste activated sludge. Black-Right-Pointing-Pointer The increased temperature demonstrated the possibility of improving typical yields of the conventional mesophilic process. Black-Right-Pointing-Pointer The temperature phased anaerobic digestion process (65 + 55 Degree-Sign C) showed the best performances with yields of 0.49 m{sup 3}/kgVS{sub fed}. Black-Right-Pointing-Pointer Ammonia and phosphate released from solids destruction determined the precipitation of struvite in the reactor. - Abstract: The paper reports the findings of a two-year pilot scale experimental trial for the mesophilic (35 Degree-Sign C), thermophilic (55 Degree-Sign C) and temperature phased (65 +more » 55 Degree-Sign C) anaerobic digestion of waste activated sludge. During the mesophilic and thermophilic runs, the reactor operated at an organic loading rate of 2.2 kgVS/m{sup 3}d and a hydraulic retention time of 20 days. In the temperature phased run, the first reactor operated at an organic loading rate of 15 kgVS/m{sup 3}d and a hydraulic retention time of 2 days while the second reactor operated at an organic loading rate of 2.2 kgVS/m{sup 3}d and a hydraulic retention time of 18 days (20 days for the whole temperature phased system). The performance of the reactor improved with increases in temperature. The COD removal increased from 35% in mesophilic conditions, to 45% in thermophilic conditions, and 55% in the two stage temperature phased system. As a consequence, the specific biogas production increased from 0.33 to 0.45 and to 0.49 m{sup 3}/kgVS{sub fed} at 35, 55, and 65 + 55 Degree-Sign C, respectively. The extreme thermophilic reactor working at 65 Degree-Sign C showed a high hydrolytic capability and a specific yield of 0.33 gCOD (soluble) per gVS{sub fed}. The effluent of the extreme thermophilic reactor showed an average concentration of soluble COD and

  19. Feasibility of thermophilic anaerobic processes for treating waste activated sludge under low HRT and intermittent mixing.

    PubMed

    Leite, Wanderli; Magnus, Bruna Scandolara; Guimarães, Lorena Bittencourt; Gottardo, Marco; Belli Filho, Paulo

    2017-10-01

    Thermophilic anaerobic digestion (AD) arises as an optimized solution for the waste activated sludge (WAS) management. However, there are few feasibility studies using low solids content typically found in the WAS, and that consider uncommon operational conditions such as intermittent mixing and low hydraulic retention time (HRT). In this investigation, a single-stage pilot reactor was used to treat WAS at low HRT (13, 9, 6 and 5 days) and intermittent mixing (withholding mixing 2 h prior feeding). Thermophilic anaerobic digestion (55 °C) was initiated from a mesophilic digester (35 °C) by the one-step startup strategy. Although instabilities on partial alkalinity (1245-3000 mgCaCO 3 /L), volatile fatty acids (1774-6421 mg/L acetic acid) and biogas production (0.21-0.09 m 3 /m 3 reactor .d) were observed, methanogenesis started to recover in 18 days. The thermophilic treatment of WAS at 13 and 9 days HRT efficiently converted VS into biogas (22 and 21%, respectively) and achieved high biogas yield (0.24 and 0.22 m 3 /kgVS fed , respectively). Intermittent mixing improved the retention of methanogens inside the reactor and reduced the washout effect even at low HRT (<9 days). The negative thermal balance found was influenced by the low solids content in the WAS (2.1% TS) and by the heat losses from the digester walls. The energy balance and economic analyses demonstrated the feasibility of thermophilic AD of WAS in a hypothetical full-scale system, when the heat energy could be recovered from methane in a scenario of higher solids concentration in the substrate (>5% TS). Copyright © 2017 Elsevier Ltd. All rights reserved.

  20. Performance optimization and validation of ADM1 simulations under anaerobic thermophilic conditions.

    PubMed

    Atallah, Nabil M; El-Fadel, Mutasem; Ghanimeh, Sophia; Saikaly, Pascal; Abou-Najm, Majdi

    2014-12-01

    In this study, two experimental sets of data each involving two thermophilic anaerobic digesters treating food waste, were simulated using the Anaerobic Digestion Model No. 1 (ADM1). A sensitivity analysis was conducted, using both data sets of one digester, for parameter optimization based on five measured performance indicators: methane generation, pH, acetate, total COD, ammonia, and an equally weighted combination of the five indicators. The simulation results revealed that while optimization with respect to methane alone, a commonly adopted approach, succeeded in simulating methane experimental results, it predicted other intermediary outputs less accurately. On the other hand, the multi-objective optimization has the advantage of providing better results than methane optimization despite not capturing the intermediary output. The results from the parameter optimization were validated upon their independent application on the data sets of the second digester. Copyright © 2014 Elsevier Ltd. All rights reserved.

  1. Mitigating ammonia inhibition of thermophilic anaerobic treatment of digested piggery wastewater: use of pH reduction, zeolite, biomass and humic acid.

    PubMed

    Ho, L; Ho, G

    2012-09-15

    High free ammonia released during anaerobic digestion of livestock wastes is widely known to inhibit methanogenic microorganisms and result in low methane production. This was encountered during our earlier thermophilic semi-continuously fed continuously-stirred tank reactor (CSTR) treatment of piggery wastewater. This study explored chemical and biological means to mitigate ammonia inhibition on thermophilic anaerobic treatment of piggery wastewater with the aim to increase organic volatile carbon reduction and methane production. A series of thermophilic anaerobic batch experiments were conducted on the digested piggery effluent to investigate the effects of pH reduction (pH 8.3 to 7.5, 7.0 and 6.5) and additions of biomass (10% v/v and 19% v/v anaerobic digested piggery biomass and aerobic-anaerobic digested municipal biomass), natural zeolite (10, 15 and 20 g/L) and humic acid (1, 5 and 10 g/L) on methane production at 55 °C for 9-11 days. Reduction of the wastewater pH from its initial pH of 8.3 to 6.5 produced the greatest stimulation of methane production (3.4 fold) coupled with reductions in free ammonia (38 fold) and total volatile fatty acids (58% TVFA), particularly acetate and propionate. Addition of 10-20 g/L zeolite to piggery wastewater with and without pH reduction to 6.5 further enhanced total VFA reduction and methane production over their respective controls, with 20 g/L zeolite producing the highest enhancement effect despite the ammonia-nitrogen concentrations of the treated wastewaters remaining high. Without pH reduction, zeolite concentration up to 20 g/L was required to achieve comparable methane enhancement as the pH-reduced wastewater at pH 6.5. Although biomass (10% v/v piggery and municipal wastes) and low humic acid (1 and 5 g/L) additions enhanced total VFA reduction and methane production, they elevated the residual effluent total COD concentrations over the control wastewaters (pH-unadjusted and pH-reduced) unlike zeolite

  2. Optimization of micronutrient supplement for enhancing biogas production from food waste in two-phase thermophilic anaerobic digestion.

    PubMed

    Menon, Ajay; Wang, Jing-Yuan; Giannis, Apostolos

    2017-01-01

    The aim of this study was to enhance the biogas productivity of two-phase thermophilic anaerobic digestion (AD) using food waste (FW) as the primary substrate. The influence of adding four trace metals (Ca, Mg, Co, and Ni) as micronutrient supplement in the methanogenic phase of the thermophilic system was investigated. Initially, Response Surface Methodology (RSM) was applied to determine the optimal concentration of micronutrients in batch experiments. The results showed that optimal concentrations of 303, 777, 7 and 3mg/L of Ca, Mg, Co and Ni, respectively, increased the biogas productivity as much as 50% and significantly reduced the processing time. The formulated supplement was tested in continuous two-phase thermophilic AD system with regard to process stability and productivity. It was found that a destabilized thermophilic AD process encountering high VFA accumulation recovered in less than two weeks, while the biogas production was improved by 40% yielding 0.46L CH 4 /gVS added /day. There was also a major increase in soluble COD utilization upon the addition of micronutrient supplement. The results of this study indicate that a micronutrient supplement containing Ca, Mg, Co and Ni could probably remedy any type of thermophilic AD process. Copyright © 2016 Elsevier Ltd. All rights reserved.

  3. Kinetics of inactivation of indicator pathogens during thermophilic anaerobic digestion.

    PubMed

    Popat, Sudeep C; Yates, Marylynn V; Deshusses, Marc A

    2010-12-01

    Thermophilic anaerobic sludge digestion is a promising process to divert waste to beneficial use, but an important question is the required temperature and holding time to achieve a given degree of pathogen inactivation. In this study, the kinetics of inactivation of Ascaris suum and vaccine strain poliovirus type 1 (PVS-1), selected as indicators for helminth ova and enteric viruses respectively, were determined during anaerobic digestion at temperatures ranging from 51 to 56 °C. Inactivation of both indicator organisms was fast with greater than two log reductions achieved within 2 h for A. suum and three log reductions for PVS-1, suggesting that the current U.S. regulations are largely conservative. The first-order inactivation rate constants k followed Arrhenius relationship with activation energies of 105 and 39 KJ mol(-1) for A. suum and PVS-1, respectively indicating that A. suum was more sensitive to temperature. Although inactivation was fast, the presence of compounds in the sludge that are known to be protective of pathogen inactivation was observed, suggesting that composition-dependent time-temperature relationships are necessary. Copyright © 2010 Elsevier Ltd. All rights reserved.

  4. Ecology and biotechnological potential of the thermophilic fermentative Coprothermobacter spp.

    PubMed

    Gagliano, M C; Braguglia, C M; Petruccioli, M; Rossetti, S

    2015-05-01

    Thermophilic bacteria have been isolated from several terrestrial, marine and industrial environments. Anaerobic digesters treating organic wastes are often an important source of these microorganisms, which catalyze a wide array of metabolic processes. Moreover, organic wastes are primarily composed of proteins, whose degradation is often incomplete. Coprothermobacter spp. are proteolytic anaerobic thermophilic microbes identified in several studies focused on the analysis of the microbial community structure in anaerobic thermophilic reactors. They are currently classified in the phylum Firmicutes; nevertheless, several authors showed that the Coprothermobacter group is most closely related to the phyla Dictyoglomi and Thermotoga. Since only a few proteolytic anaerobic thermophiles have been characterized so far, this microorganism has attracted the attention of researchers for its potential applications with high-temperature environments. In addition to proteolysis, Coprothermobacter spp. showed several metabolic abilities and may have a biotechnological application either as source of thermostable enzymes or as inoculum in anaerobic processes. Moreover, they can improve protein degradation by establishing a syntrophy with hydrogenotrophic archaea. To gain a better understanding of the phylogenesis, metabolic capabilities and adaptations of these microorganisms, it is of importance to better define the role in thermophilic environments and to disclose properties not yet investigated. © FEMS 2015. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

  5. Ethanol Production from Wet-Exploded Wheat Straw Hydrolysate by Thermophilic Anaerobic Bacterium Thermoanaerobacter BG1L1 in a Continuous Immobilized Reactor

    NASA Astrophysics Data System (ADS)

    Georgieva, Tania I.; Mikkelsen, Marie J.; Ahring, Birgitte K.

    Thermophilic ethanol fermentation of wet-exploded wheat straw hydrolysate was investigated in a continuous immobilized reactor system. The experiments were carried out in a lab-scale fluidized bed reactor (FBR) at 70°C. Undetoxified wheat straw hydrolysate was used (3-12% dry matter), corresponding to sugar mixtures of glucose and xylose ranging from 12 to 41 g/1. The organism, thermophilic anaerobic bacterium Thermoanaerobacter BG1L1, exhibited significant resistance to high levels of acetic acid (up to 10 g/1) and other metabolic inhibitors present in the hydrolysate. Although the hydrolysate was not detoxified, ethanol yield in a range of 0.39-0.42 g/g was obtained. Overall, sugar efficiency to ethanol was 68-76%. The reactor was operated continuously for approximately 143 days, and no contamination was seen without the use of any agent for preventing bacterial infections. The tested microorganism has considerable potential to be a novel candidate for lignocellulose bioconversion into ethanol. The work reported here also demonstrates that the use of FBR configuration might be a viable approach for thermophilic anaerobic ethanol fermentation.

  6. Anaerobic co-digestion of steam-treated Quercus serrata chips and sewage sludge under mesophilic and thermophilic conditions.

    PubMed

    Wang, Feng; Hidaka, Taira; Sakurai, Kensuke; Tsumori, Jun

    2014-08-01

    The biodegradation of Quercus serrata chips was evaluated by anaerobic digestion under various steam explosion conditions. In continuous experiments, untreated chips (W₀) and chips steam-treated at less than 1.0 MPa (W₁) and 2.0 MPa (W₄) were co-digested with sewage sludge (S₁ and S₂) taken from two different wastewater treatment plants. The apparent methane yield of W₁ and W₄ co-digested with S₁ (thermophilic) was 261 dm(3)/kgVS (volatile solids) and 248 dm(3)/kgVS, respectively. The apparent methane yield of W₄ co-digested with S₂ was 258 dm(3)/kgVS (mesophilic) and 271 dm(3)/kgVS (thermophilic). Methane production was inhibited by W₀ due to components released during hydrolysis. The methane conversion ratio of pretreated chips obtained in batch experiments varied from 40.5% to 53.8% (mesophilic) and from 49.0% to 63.7% (thermophilic). The methane conversion ratio increased with decreasing acid-soluble lignin content in the chips. Copyright © 2014 Elsevier Ltd. All rights reserved.

  7. Screening of thermophilic anaerobic bacteria for solid substrate cultivation on lignocellulosic substrates.

    PubMed

    Chinn, Mari S; Nokes, Sue E; Strobel, Herbert J

    2006-01-01

    Interest in solid substrate cultivation (SSC) techniques is gaining for biochemical production from renewable resources; however, heat and mass transfer problems may limit application of this technique. The use of anaerobic thermophiles in SSC offers a unique solution to overcoming these challenges. The production potential of nine thermophilic anaerobic bacteria was examined on corn stover, sugar cane bagasse, paper pulp sludge, and wheat bran in submerged liquid cultivation (SmC) and SSC. Production of acetate, ethanol, and lactate was measured over a 10 day period, and total product concentrations were used to compare the performance of different organism-substrate combinations using the two cultivation methods. Overall microbial activity in SmC and SSC was dependent on the organism and growth substrate. Clostridium thermocellum strains JW20, LQRI, and 27405 performed significantly better in SSC when grown on sugar cane bagasse and paper pulp sludge, producing at least 70 and 170 mM of total products, respectively. Growth of C. thermocellum strains in SSC on paper pulp sludge proved to be most favorable, generating at least twice the concentration of total products produced in SmC (p-value < 0.05). Clostridium thermolacticum TC21 demonstrated growth on all substrates producing 30-80 and 60-116 mM of total product in SmC and SSC, respectively. Bacterial species with optimal growth temperatures of 70 degrees C grew best on wheat bran in SmC, producing total product concentrations of 45-75 mM. For some of the organism-substrate combinations total end product concentrations in SSC exceeded those in SmC, indicating that SSC may be a promising alternative for microbial activity and value-added biochemical production.

  8. Mesophilic and thermophilic anaerobic co-digestion of abattoir wastewater and fruit and vegetable waste in anaerobic sequencing batch reactors.

    PubMed

    Bouallagui, Hassib; Rachdi, Boutheina; Gannoun, Hana; Hamdi, Moktar

    2009-06-01

    Anaerobic co-digestion of fruit and vegetable waste (FVW) and abattoir wastewater (AW) was investigated using anaerobic sequencing batch reactors (ASBRs). The effects of hydraulic retention time (HRT) and temperature variations on digesters performances were examined. At both 20 and 10 days biogas production for co-digestion was greater thanks to the improved balance of nutrients. The high specific gas productions for the different digestion processes were 0.56, 0.61 and 0.85 l g(-1) total volatile solids (TVS) removal for digesters treating AW, FVW and AW + FVW, respectively. At an HRT of 20 days, biogas production rates from thermophilic digesters were higher on average than from mesophilic AW, FVW and AW + FVW digestion by 28.5, 44.5 and 25%, respectively. However, at 10 days of HRT results showed a decrease of biogas production rate for AW and AW + FVW digestion processes due to the high amount of free ammonia at high organic loading rate (OLR).

  9. Escherichia coli inactivation kinetics in anaerobic digestion of dairy manure under moderate, mesophilic and thermophilic temperatures

    PubMed Central

    2011-01-01

    Batch anaerobic digestion experiments using dairy manure as feedstocks were performed at moderate (25°C), mesophilic (37°C), and thermophilic (52.5°C) temperatures to understand E. coli, an indicator organism for pathogens, inactivation in dairy manure. Incubation periods at 25, 37, and 52.5°C, were 61, 41, and 28 days respectively. Results were used to develop models for predicting E. coli inactivation and survival in anaerobic digestion. For modeling we used the decay of E. coli at each temperature to calculate the first-order inactivation rate coefficients, and these rates were used to formulate the time - temperature - E. coli survival relationships. We found the inactivation rate coefficient at 52.5°C was 17 and 15 times larger than the inactivation rate coefficients at 25 and 37°C, respectively. Decimal reduction times (D10; time to achieve one log removal) at 25, 37, and 52.5°C, were 9 -10, 7 - 8 days, and < 1 day, respectively. The Arrhenius correlation between inactivation rate coefficients and temperatures over the range 25 -52.5°C was developed to understand the impacts of temperature on E. coli inactivation rate. Using this correlation, the time - temperature - E. coli survival relationships were derived. Besides E. coli inactivation, impacts of temperature on biogas production, methane content, pH change, ORP, and solid reduction were also studied. At higher temperatures, biogas production and methane content was greater than that at low temperatures. While at thermophilic temperature pH was increased, at mesophilic and moderate temperatures pH were reduced over the incubation period. These results can be used to understand pathogen inactivation during anaerobic digestion of dairy manure, and impacts of temperatures on performance of anaerobic digesters treating dairy manure. PMID:21906374

  10. Anaerobic co-digestion of food waste and chemically enhanced primary-treated sludge under mesophilic and thermophilic conditions.

    PubMed

    Obulisamy, Parthiba Karthikeyan; Chakraborty, Debkumar; Selvam, Ammaiyappan; Wong, Jonathan W C

    2016-12-01

    Anaerobic co-digestion of food waste with primary sewage sludge is beneficial for urban centers, while the optimized conditions reported in the literature are not locally suitable for Hong Kong. Therefore, the present study was aimed to develop an optimized mixing ratio of food waste to chemically enhanced primary-treated sewer sludge (CEPT) for co-digestion using batch tests under mesophilic (37°C) and thermophilic (55°C) conditions. The mixing ratios of 1:1, 1:2, 1:3, 2:1 and 3:1 (v v(-1)) of food waste to CEPT sludge was tested under the following conditions: temperature - 35°C and 55°C; pH - not regulated; agitation - 150 rpm and time - 20 days. The thermophilic incubations led a good hydrolysis rate and 2-12-fold higher enzyme activities than in mesophilic incubations for different mixing ratios. While the acidogenesis were found retarded that leading to 'sour and stuck' digestion for all mixing ratio of food waste to CEPT sludge from thermophilic incubations. The measured zeta potential was most favourable (-5 to -16.8 mV) for methane production under thermophilic incubations; however the CH4 recovery was less than that in mesophilic incubations. The results suggested that the quick hydrolysis and subsequent acid accumulation under thermophilic incubation lead to inhibited methanogenesis at the early stage than in mesophilic systems. It is concluded that buffer addition is therefore required for any mixing ratio of food waste to CEPT sludge for improved CH4 recovery for both mesophilic and thermophilic operations.

  11. Caldanaerobacter uzonensis sp. nov., an anaerobic, thermophilic, heterotrophic bacterium isolated from a hot spring.

    PubMed

    Kozina, Irina V; Kublanov, Ilya V; Kolganova, Tatyana V; Chernyh, Nikolai A; Bonch-Osmolovskaya, Elizaveta A

    2010-06-01

    An anaerobic thermophilic bacterium, strain K67(T), was isolated from a terrestrial hot spring of Uzon Caldera, Kamchatka Peninsula. Analysis of the 16S rRNA gene sequence revealed that the novel isolate belongs to the genus Caldanaerobacter, with 95 % 16S rRNA gene sequence similarity to Caldanaerobacter subterraneus subsp. subterraneus SEBR 7858(T), suggesting that it represents a novel species of the genus Caldanaerobacter. Strain K67(T) was characterized as an obligate anaerobe, a thermophile (growth at 50-75 degrees capital ES, Cyrillic; optimum 68-70 degrees C), a neutrophile (growth at pH(25 degrees C) 4.8-8.0; optimum pH(25 degrees C) 6.8) and an obligate organotroph (growth by fermentation of various sugars, peptides and polysaccharides). Major fermentation products were acetate, H2 and CO2; ethanol, lactate and l-alanine were formed in smaller amounts. Thiosulfate stimulated growth and was reduced to hydrogen sulfide. Nitrate, sulfate, sulfite and elemental sulfur were not reduced and did not stimulate growth. Thus, according to the strain's phylogenetic position and phenotypic novelties (lower upper limit of temperature range for growth, the ability to grow on arabinose, the inability to reduce elemental sulfur and the formation of alanine as a minor fermentation product), the novel species Caldanaerobacter uzonensis sp. nov. is proposed, with the type strain K67(T) (=DSM 18923(T) =VKM capital VE, Cyrillic-2408(T)).

  12. Conductive iron oxides accelerate thermophilic methanogenesis from acetate and propionate.

    PubMed

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

    2015-06-01

    Anaerobic digester is one of the attractive technologies for treatment of organic wastes and wastewater, while continuous development and improvements on their stable operation with efficient organic removal are required. Particles of conductive iron oxides (e.g., magnetite) are known to facilitate microbial interspecies electron transfer (termed as electric syntrophy). Electric syntrophy has been reported to enhance methanogenic degradation of organic acids by mesophilic communities in soil and anaerobic digester. Here we investigated the effects of supplementation of conductive iron oxides (magnetite) on thermophilic methanogenic microbial communities derived from a thermophilic anaerobic digester. Supplementation of magnetite accelerated methanogenesis from acetate and propionate under thermophilic conditions, while supplementation of ferrihydrite also accelerated methanogenesis from propionate. Microbial community analysis revealed that supplementation of magnetite drastically changed bacterial populations in the methanogenic acetate-degrading cultures, in which Tepidoanaerobacter sp. and Coprothermobacter sp. dominated. These results suggest that supplementation of magnetite induce electric syntrophy between organic acid-oxidizing bacteria and methanogenic archaea and accelerate methanogenesis even under thermophilic conditions. Findings from this study would provide a possibility for the achievement of stably operating thermophilic anaerobic digestion systems with high efficiency for removal of organics and generation of CH4. Copyright © 2014 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.

  13. [Effect of NaOH-treatment on dry-thermophilic anaerobic digestion of Spartina alterniflora].

    PubMed

    Chen, Guang-Yin; Zheng, Zheng; Chang, Zhi-Zhou; Ye, Xiao-Mei; Luo, Yan

    2011-07-01

    In order to improve the biotransformation rate of lignocellulosic materials, sodium hydroxide (NaOH) was widely used to pretreat lignocellulosic materials. Effect of NaOH-treatment on dry-thermophilic anaerobic digestion of Spartina alternflora was studied by batch model under the temperature of 55 degrees C +/- 1 degrees C, at the initial total solid loading (TSL) of 20%. The results indicated that biogas production was inhibited by NaOH-treatment and improved by NaOH-treatment with water washed. The cumulative biogas yield of control (CK), NaOH-treated and NaOH-treated with water washed (NaOH + water) were 268.35 mL/g, 205.76 mL/g and 299.97 mL/g, respectively. The methane content of CK and NaOH + water treatments kept stable while fluctuation of NaOH-treated treatment during anaerobic digestion process was observed. Compared with CK and NaOH + water treatments, methane content of NaOH-treated treatment was improved by 5.30%. The content of hemi-cellulose and cellulose of S. alternifora decreased while content of lignin of S. alterniflora increased after 51-day anaerobic digestion. The crystallinity of cellulose of S. alterniflora increased after NaOH-treatment which was consistent to the result of FTIR. The lignocellulosic structure was destroyed and the biodegradability of S. alterniflora was increased by NaOH pretreatment. However, the amount of Na+ was taken into the anaerobic system, besides the high Na+ content in the plant itself which inhibited the anaerobic microorganisms. Therefore, NaOH-treatment is considered to be unsuitable for the anaerobic digestion of S. alterniflora.

  14. Thermophilic and hyper-thermophilic co-digestion of waste activated sludge and fat, oil and grease: Evaluating and modeling methane production.

    PubMed

    Alqaralleh, Rania Mona; Kennedy, Kevin; Delatolla, Robert; Sartaj, Majid

    2016-12-01

    Renewable energy and clean environment are two crucial requirements for our modern world. Low cost, energy production and limited environmental impact make anaerobic digestion (AD) a promising technology for stabilizing organic waste and in particular, sewage waste. The anaerobic co-digestion of thickened waste activated sludge (TWAS) and sewage treatment plant trapped fat, oil and grease (FOG) using different FOG-TWAS mixtures (20, 40, 60 and 80% of FOG based on total volatile solids (TVS)) were investigated in this study using both thermophilic (55 ± 1 °C) and two stages hyper-thermophilic/thermophilic (70 ± 1 °C and 55 ± 1 °C) anaerobic co-digestion. The hyper-thermophilic co-digestion approach as a part of the co-digestion process has been shown to be very useful in improving the methane production. During hyper-thermophilic biochemical methane potential (BMP) assay testing the sample with 60% FOG (based on TVS) has been shown to significantly increase the maximum methane production to 673.1 ± 14.0 ml of methane as compared to 316.4 ± 14.3 ml of methane for the control sample. This represents a 112.7% increase in methane production compared to the control sample considered in this paper. These results signify the importance of hyper-thermophilic digestion to the co-digestion of TWAS-FOG field. Copyright © 2016 Elsevier Ltd. All rights reserved.

  15. Isolation and characterization of two novel ethanol-tolerant facultative-anaerobic thermophilic bacteria strains from waste compost.

    PubMed

    Fong, Jiunn C N; Svenson, Charles J; Nakasugi, Kenlee; Leong, Caine T C; Bowman, John P; Chen, Betty; Glenn, Dianne R; Neilan, Brett A; Rogers, Peter L

    2006-10-01

    In a search for potential ethanologens, waste compost was screened for ethanol-tolerant thermophilic microorganisms. Two thermophilic bacterial strains, M5EXG and M10EXG, with tolerance of 5 and 10% (v/v) ethanol, respectively, were isolated. Both isolates are facultative anaerobic, non-spore forming, non-motile, catalase-positive, oxidase-negative, Gram-negative rods that are capable of utilizing a range of carbon sources including arabinose, galactose, mannose, glucose and xylose and produce low amounts of ethanol, acetate and lactate. Growth of both isolates was observed in fully defined minimal media within the temperature range 50-80 degrees C and pH 6.0-8.0. Phylogenetic analysis of the 16S rDNA sequences revealed that both isolates clustered with members of subgroup 5 of the genus Bacillus. G+C contents and DNA-DNA relatedness of M5EXG and M10EXG revealed that they are strains belonging to Geobacillus thermoglucosidasius. However, physiological and biochemical differences were evident when isolates M5EXG and M10EXG were compared with G. thermoglucosidasius type strain (DSM 2542(T)). The new thermophilic, ethanol-tolerant strains of G. thermoglucosidasius may be candidates for ethanol production at elevated temperatures.

  16. Anaerobic digestion of whole stillage from dry-grind corn ethanol plant under mesophilic and thermophilic conditions.

    PubMed

    Eskicioglu, Cigdem; Kennedy, Kevin J; Marin, Juan; Strehler, Benjamin

    2011-01-01

    Anaerobic digestion of whole stillage from a dry-grind corn-based ethanol plant was evaluated by batch and continuous-flow digesters under thermophilic and mesophilic conditions. At whole corn stillage concentrations of 6348 to 50,786 mg total chemical oxygen demand (TCOD)/L, at standard temperature (0 °C) and pressure (1 atm), preliminary biochemical methane potential assays produced 88±8 L (49±5 L CH4) and 96±19 L (65±14 L CH4) biogas per L stillage from mesophilic and thermophilic digesters, respectively. Continuous-flow studies for the full-strength stillage (TCOD=254 g/L) at organic loadings of 4.25, 6.30 and 9.05 g TCOD/L days indicated unstable performance for the thermophilic digester. Among the sludge retention times (SRTs) of 60, 45 and 30 days tested, the mesophilic digestion was successful only at 60 days-SRT which does not represent a practical operation time for a large scale bioethanol plant. Future laboratory studies will focus on different reactor configurations to reduce the SRT needed in the digesters. Copyright © 2010 Elsevier Ltd. All rights reserved.

  17. Combined thermophilic aerobic process and conventional anaerobic digestion: effect on sludge biodegradation and methane production.

    PubMed

    Dumas, C; Perez, S; Paul, E; Lefebvre, X

    2010-04-01

    The efficiency of hyper-thermophilic (65 degrees Celsius) aerobic process coupled with a mesophilic (35 degrees Celsius) digester was evaluated for the activated sludge degradation and was compared to a conventional mesophilic digester. For two Sludge Retention Time (SRT), 21 and 42 days, the Chemical Oxygen Demand (COD) solubilisation and biodegradation processes, the methanisation yield and the aerobic oxidation were investigated during 180 days. The best results were obtained at SRT of 44 days; the COD removal yield was 30% higher with the Mesophilic Anaerobic Digestion/Thermophilic Aerobic Reactor (MAD-TAR) co-treatment. An increase of the sludge intrinsic biodegradability is also observed (20-40%), showing that the unbiodegradable COD in mesophilic conditions becomes bioavailable. However, the methanisation yield was quite similar for both processes at a same SRT. Finally, such a process enables to divide by two the volume of digester with an equivalent efficiency. Copyright 2009 Elsevier Ltd. All rights reserved.

  18. Effect of moisture of municipal biowaste on start-up and efficiency of mesophilic and thermophilic dry anaerobic digestion.

    PubMed

    Li, Chaoran; Mörtelmaier, Christoph; Winter, Josef; Gallert, Claudia

    2014-09-01

    Methane production from biowaste with 20-30% dry matter (DM) by box-type dry anaerobic digestion and contributing bacteria were determined for incubation at 20, 37 and 55 °C. The same digestion efficiency as for wet anaerobic digestion of biowaste was obtained for dry anaerobic digestion with 20% DM content at 20, 37 and 55 °C and with 25% DM content at 37 and 55 °C. No or only little methane was produced in dry anaerobic reactors with 30% DM at 20, 37 or 55 °C. Population densities in the 20-30% DM-containing biowaste reactors were similar although in mesophilic and thermophilic biowaste reactors with 30% DM content significantly less but phylogenetically more diverse archaea existed. Biogas production in the 20% and 25% DM assays was catalyzed by Methanosarcinales and Methanomicrobiales. In all assays Pelotomaculum and Syntrophobacter species were dominant propionate degraders. Copyright © 2014 Elsevier Ltd. All rights reserved.

  19. Evolution of microorganisms in thermophilic-dry anaerobic digestion.

    PubMed

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

    2008-05-01

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

  20. Changes of resistome, mobilome and potential hosts of antibiotic resistance genes during the transformation of anaerobic digestion from mesophilic to thermophilic.

    PubMed

    Tian, Zhe; Zhang, Yu; Yu, Bo; Yang, Min

    2016-07-01

    This study aimed to reveal how antibiotic resistance genes (ARGs) and their horizontal and vertical transfer-related items (mobilome and bacterial hosts) respond to the transformation of anaerobic digestion (AD) from mesophilic to thermophilic using one-step temperature increase. The resistomes and mobilomes of mesophilic and thermophilic sludge were investigated using metagenome sequencing, and the changes in 24 representative ARGs belonging to three categories, class 1 integron and bacterial genera during the transition period were further followed using quantitative PCR and 454-pyrosequencing. After the temperature increase, resistome abundance in the digested sludge decreased from 125.97 ppm (day 0, mesophilic) to 50.65 ppm (day 57, thermophilic) with the reduction of most ARG types except for the aminoglycoside resistance genes. Thermophilic sludge also had a smaller mobilome, including plasmids, insertion sequences and integrons, than that of mesophilic sludge, suggesting the lower horizontal transfer potential of ARGs under thermophilic conditions. On the other hand, the total abundance of 18 bacterial genera, which were suggested as the possible hosts for 13 ARGs through network analysis, decreased from 23.27% in mesophilic sludge to 11.92% in thermophilic sludge, indicating fewer hosts for the vertical expansion of ARGs after the increase in temperature. These results indicate that the better reduction of resistome abundance by thermophilic AD might be associated with the decrease of both the horizontal and vertical transferability of ARGs. Copyright © 2016 Elsevier Ltd. All rights reserved.

  1. Combined mesophilic anaerobic and thermophilic aerobic digestion process: effect on sludge degradation and variation of sludge property.

    PubMed

    Cheng, Jiehong; Ji, Yuehong; Kong, Feng; Chen, Xian

    2013-12-01

    One-stage autothermal thermophilic aerobic digestion (ATAD) is effective for the reduction of volatile solids (VSs) and pathogen in sewage sludges. A novel process of combining mesophilic (<35 °C) anaerobic digestion with a thermophilic (55 °C) aerobic digestion process (AN/TAD) occurred in a one-stage digester, which was designed for aeration energy savings. The efficiency of sludge degradation and variation of sludge properties by batch experiments were evaluated for the AN/TAD digester with an effective volume of 23 L for 30 days compared with conventional thermophilic aerobic digestion (TAD). The AN/TAD system can efficiently achieve sludge stabilization on the 16th day with a VS removal rate of 38.1 %. The AN/TAD system was operated at lower ORP values in a digestion period with higher contents of total organic compounds, volatile fatty acids, protein, and polysaccharide in the soluble phase than those of the TAD system, which can rapidly decreased and had low values in the late period of digestion for the AN/TAD system. In the AN/TAD system, intracellular substances had lysis because of initial hydrolytic acidification.

  2. The role of zero valent iron on the fate of tetracycline resistance genes and class 1 integrons during thermophilic anaerobic co-digestion of waste sludge and kitchen waste.

    PubMed

    Gao, Pin; Gu, Chaochao; Wei, Xin; Li, Xiang; Chen, Hong; Jia, Hanzhong; Liu, Zhenhong; Xue, Gang; Ma, Chunyan

    2017-03-15

    Activated sludge has been identified as a potential significant source of antibiotic resistance genes (ARGs) to the environment. Anaerobic digestion is extensively used for sludge stabilization and resource recovery, and represents a crucial process for controlling the dissemination of ARGs prior to land application of digested sludge. The objective of this study is to investigate the effect of zero valent iron (Fe 0 ) on the attenuation of seven representative tetracycline resistance genes (tet, tet(A), tet(C), tet(G), tet(M), tet(O), tet(W), and tet(X)), and the integrase gene intI1 during thermophilic anaerobic co-digestion of waste sludge and kitchen waste. Significant decrease (P < 0.05) in the quantities of tet (except tet(W)) and intI1 genes was observed at Fe 0 dosage of 5 g/L, whereas no significant differences (P > 0.05) were found for all gene targets between digesters with Fe 0 dosages of 5 and 60 g/L. A first-order kinetic model favorably described the trends in concentrations of tet and intI1 gene targets during thermophilic anaerobic digestion with or without Fe 0 . Notably, tet genes encoding different resistance mechanisms behaved distinctly in anaerobic digesters, although addition of Fe 0 could enhance their reduction. The overall results of this research suggest that thermophilic anaerobic digestion with Fe 0 can be a potential alternative technology for the attenuation of tet and intI1 genes in waste sludge. Copyright © 2016 Elsevier Ltd. All rights reserved.

  3. Bacterial cellulose hydrolysis in anaerobic environmental subsystems--Clostridium thermocellum and Clostridium stercorarium, thermophilic plant-fiber degraders.

    PubMed

    Zverlov, Vladimir V; Schwarz, Wolfgang H

    2008-03-01

    Cellulose degradation is a rare trait in bacteria. However, the truly cellulolytic bacteria are extremely efficient hydrolyzers of plant cell wall polysaccharides, especially those in thermophilic anaerobic ecosystems. Clostridium stercorarium, a thermophilic ubiquitous soil dweller, has a simple cellulose hydrolyzing enzyme system of only two cellulases. However, it seems to be better suited for the hydrolysis of a wide range of hemicelluloses. Clostridium thermocellum, an ubiquitous thermophilic gram-type positive bacterium, is one of the most successful cellulose degraders known. Its extracellular enzyme complex, the cellulosome, was prepared from C. thermocellum cultures grown on cellulose, cellobiose, barley beta-1,3-1,4-glucan, or a mixture of xylan and cellulose. The single proteins were identified by peptide chromatography and MALDI-TOF-TOF. Eight cellulosomal proteins could be found in all eight preparations, 32 proteins occur in at least one preparation. A number of enzymatic components had not been identified previously. The proportion of components changes if C. thermocellum is grown on different substrates. Mutants of C. thermocellum, devoid of scaffoldin CipA, that now allow new types of experiments with in vitro cellulosome reassembly and a role in cellulose hydrolysis are described. The characteristics of these mutants provide strong evidence of the positive effect of complex (cellulosome) formation on hydrolysis of crystalline cellulose.

  4. Modeling temperature variations in a pilot plant thermophilic anaerobic digester.

    PubMed

    Valle-Guadarrama, Salvador; Espinosa-Solares, Teodoro; López-Cruz, Irineo L; Domaschko, Max

    2011-05-01

    A model that predicts temperature changes in a pilot plant thermophilic anaerobic digester was developed based on fundamental thermodynamic laws. The methodology utilized two simulation strategies. In the first, model equations were solved through a searching routine based on a minimal square optimization criterion, from which the overall heat transfer coefficient values, for both biodigester and heat exchanger, were determined. In the second, the simulation was performed with variable values of these overall coefficients. The prediction with both strategies allowed reproducing experimental data within 5% of the temperature span permitted in the equipment by the system control, which validated the model. The temperature variation was affected by the heterogeneity of the feeding and extraction processes, by the heterogeneity of the digestate recirculation through the heating system and by the lack of a perfect mixing inside the biodigester tank. The use of variable overall heat transfer coefficients improved the temperature change prediction and reduced the effect of a non-ideal performance of the pilot plant modeled.

  5. Development of bioconcrete material using an enrichment culture of novel thermophilic anaerobic bacteria.

    PubMed

    Ghosh, P; Mandal, S; Pal, S; Bandyopadhyaya, G; Chattopadhyay, B D

    2006-04-01

    In the biosphere, bacteria can function as geo-chemical agents, promoting the dispersion, fractionation and/or concentration of materials. Microbial mineral precipitation is resulted from metabolic activities of microorganisms. Based on this biomineralogy concept, an attempt has been made to develop bioconcrete material incorporating of an enrichment culture of thermophilic and anaerobic bacteria within cement-sand mortar/concrete. The results showed a significant increase in compressive strength of both cement-sand mortar and concrete due to the development of filler material within the pores of cement sand matrix. Maximum strength was observed at concentration 10(5)cell/ml of water used in mortar/concrete. Addition of Escherichia coil or media composition on mortar showed no such improvement in strength.

  6. Anaerobic digestion of nitrogen rich poultry manure: Impact of thermophilic biogas process on metal release and microbial resistances.

    PubMed

    Anjum, Reshma; Grohmann, Elisabeth; Krakat, Niclas

    2017-02-01

    Poultry manure is a nitrogen rich fertilizer, which is usually recycled and spread on agricultural fields. Due to its high nutrient content, chicken manure is considered to be one of the most valuable animal wastes as organic fertilizer. However, when chicken litter is applied in its native form, concerns are raised as such fertilizers also include high amounts of antibiotic resistant pathogenic Bacteria and heavy metals. We studied the impact of an anaerobic thermophilic digestion process on poultry manure. Particularly, microbial antibiotic resistance profiles, mobile genetic elements promoting the resistance dissemination in the environment as well as the presence of heavy metals were focused in this study. The initiated heat treatment fostered a community shift from pathogenic to less pathogenic bacterial groups. Phenotypic and molecular studies demonstrated a clear reduction of multiple resistant pathogens and self-transmissible plasmids in the heat treated manure. That treatment also induced a higher release of metals and macroelements. Especially, Zn and Cu exceeded toxic thresholds. Although the concentrations of a few metals reached toxic levels after the anaerobic thermophilic treatment, the quality of poultry manure as organic fertilizer may raise significantly due to the elimination of antibiotic resistance genes (ARG) and self-transmissible plasmids. Copyright © 2016 Elsevier Ltd. All rights reserved.

  7. Thermophilic versus Mesophilic Anaerobic Digestion of Sewage Sludge: A Comparative Review

    PubMed Central

    Gebreeyessus, Getachew D.; Jenicek, Pavel

    2016-01-01

    During advanced biological wastewater treatment, a huge amount of sludge is produced as a by-product of the treatment process. Hence, reuse and recovery of resources and energy from the sludge is a big technological challenge. The processing of sludge produced by Wastewater Treatment Plants (WWTPs) is massive, which takes up a big part of the overall operational costs. In this regard, anaerobic digestion (AD) of sewage sludge continues to be an attractive option to produce biogas that could contribute to the wastewater management cost reduction and foster the sustainability of those WWTPs. At the same time, AD reduces sludge amounts and that again contributes to the reduction of the sludge disposal costs. However, sludge volume minimization remains, a challenge thus improvement of dewatering efficiency is an inevitable part of WWTP operation. As a result, AD parameters could have significant impact on sludge properties. One of the most important operational parameters influencing the AD process is temperature. Consequently, the thermophilic and the mesophilic modes of sludge AD are compared for their pros and cons by many researchers. However, most comparisons are more focused on biogas yield, process speed and stability. Regarding the biogas yield, thermophilic sludge AD is preferred over the mesophilic one because of its faster biochemical reaction rate. Equally important but not studied sufficiently until now was the influence of temperature on the digestate quality, which is expressed mainly by the sludge dewateringability, and the reject water quality (chemical oxygen demand, ammonia nitrogen, and pH). In the field of comparison of thermophilic and mesophilic digestion process, few and often inconclusive research, unfortunately, has been published so far. Hence, recommendations for optimized technologies have not yet been done. The review presented provides a comparison of existing sludge AD technologies and the gaps that need to be filled so as to optimize

  8. Foaming phenomenon in bench-scale anaerobic digesters.

    PubMed

    Siebels, Amanda M; Long, Sharon C

    2013-04-01

    The Madison Metropolitan Sewerage District (The District) in Madison, Wisconsin has been experiencing seasonal foaming in their anaerobic biosolids digesters, which has occurred from mid-November to late June for the past few years. The exact cause(s) of foaming is unknown. Previous research findings are unclear as to whether applications of advanced anaerobic digestion processes reduce the foaming potential of digesters. The object of this study was to investigate how configurations of thermophilic and acid phase-thermophilic anaerobic digestion would affect foaming at the bench-scale level compared to single stage mesophilic digestion for The District. Bench-scale anaerobic digesters were fed with a 4 to 4.5% by dry weight of solids content blend of waste activated sludge (WAS) and primary sludge from The District. Foaming potential was monitored using Alka-Seltzer and aeration foaming tests. The bench-scale acid phase-thermophilic digester had a higher foaming potential than the bench-scale mesophilic digester. These results indicate that higher temperatures increase the foaming potential of the bench-scale anaerobic digesters. The bench-scale acid phase-thermophilic digesters had a greater percent (approximately 5 to 10%) volatile solids destruction and a greater percent (approximately 5 to 10%) total solids destruction when compared to the bench-scale mesophilic digester. Overall, for the full-scale foaming experienced by The District, it appears that adding an acid phase or switching to thermophilic digestion would not alleviate The District's foaming issues.

  9. Bio-hydrolysis and bio-hydrogen production from food waste by thermophilic and hyperthermophilic anaerobic process.

    PubMed

    Algapani, Dalal E; Qiao, Wei; Su, Min; di Pumpo, Francesca; Wandera, Simon M; Adani, Fabrizio; Dong, Renjie

    2016-09-01

    High-temperature pretreatment plays a key role in the anaerobic digestion of food waste (FW). However, the suitable temperature is not yet determined. In this work, a long-term experiment was conducted to compare hydrolysis, acidogenesis, acetogenesis, and hydrogen production at 55°C and 70°C, using real FW in CSTR reactors. The results obtained indicated that acidification was the rate-limiting step at both temperatures with similar process kinetics characterizations. However, the thermophilic pretreatment was more advantageous than the hyperthermophilic with suspended solids solubilization of 47.7% and 29.5% and total VFA vs. soluble COD ratio of 15.2% and 4.9%, for thermophilic and hyperthermophilic treatment, respectively, with a hydrolytic reaction time (HRT) of 10days and an OLR of 14kgCOD/m(3)d. Moreover, stable hydrogen yield (70.7ml-H2/gVSin) and content in off gas (58.6%) was achieved at HRT 5days, pH 5.5, and temperature of 55°C, as opposed to 70°C. Copyright © 2016 Elsevier Ltd. All rights reserved.

  10. Establishment of thermophilic anaerobic terephthalic acid degradation system through one-step temperature increase startup strategy - Revealed by Illumina Miseq Sequencing.

    PubMed

    Ma, Kai-Li; Li, Xiang-Kun; Wang, Ke; Meng, Ling-Wei; Liu, Gai-Ge; Zhang, Jie

    2017-10-01

    Over recent years, thermophilic digestion was constantly focused owing to its various advantage over mesophilic digestion. Notably, the startup approach of thermophilic digester needs to be seriously considered as unsuitable startup ways may result in system inefficiency. In this study, one-step temperature increase startup strategy from 37 °C to 55 °C was applied to establish a thermophilic anaerobic system treating terephthalic acid (TA) contained wastewater, meanwhile, the archaeal and bacterial community compositions at steady periods of 37 °C and 55 °C during the experimental process was also compared using Illumina Miseq Sequencing. The process operation demonstrated that the thermophilic TA degradation system was successfully established at 55 °C with over 95% COD reduction. For archaea community, the elevation of operational temperature from 37 °C to 55 °C accordingly increase the enrichment of hydrogenotrophic methanogens but decrease the abundance of the acetotrophic ones. While for bacterial community, the taxonomic analysis suggested that Syntrophorhabdus (27.40%) was the dominant genus promoting the efficient TA degradation under mesophilic condition, whereas OPB95 (24.99%) and TA06 (14.01%) related populations were largely observed and probably take some crucial role in TA degradation under thermophilic condition. Copyright © 2017 Elsevier Ltd. All rights reserved.

  11. Thermophilic degradation of cellulosic biomass

    NASA Astrophysics Data System (ADS)

    Ng, T.; Zeikus, J. G.

    1982-12-01

    The conversion of cellulosic biomass to chemical feedstocks and fuel by microbial fermentation is an important objective of developing biotechnology. Direct fermentation of cellulosic derivatives to ethanol by thermophilic bacteria offers a promising approach to this goal. Fermentations at elevated temperatures lowers the energy demand for cooling and also facilitates the recovery of volatile products. In addition, thermophilic microorganisms possess enzymes with greater stability than those from mesophilic microorganisms. Three anaerobic thermophilic cocultures that ferment cellulosic substrate mainly to ethanol have been described: Clostridium thermocellum/Clostriidium thermohydrosulfuricum, C. thermocellum/Clostridium thermosaccharolyticum, and C. thermocellum/Thermoanaerobacter ethanolicus sp. nov. The growth characteristics and metabolic features of these cocultures are reviewed.

  12. Modified ADM1 disintegration/hydrolysis structures for modeling batch thermophilic anaerobic digestion of thermally pretreated waste activated sludge.

    PubMed

    Ramirez, Ivan; Mottet, Alexis; Carrère, Hélène; Déléris, Stéphane; Vedrenne, Fabien; Steyer, Jean-Philippe

    2009-08-01

    Anaerobic digestion disintegration and hydrolysis have been traditionally modeled according to first-order kinetics assuming that their rates do not depend on disintegration/hydrolytic biomass concentrations. However, the typical sigmoid-shape increase in time of the disintegration/hydrolysis rates cannot be described with first-order models. For complex substrates, first-order kinetics should thus be modified to account for slowly degradable material. In this study, a slightly modified IWA ADM1 model is presented to simulate thermophilic anaerobic digestion of thermally pretreated waste activated sludge. Contois model is first included for disintegration and hydrolysis steps instead of first-order kinetics and Hill function is then used to model ammonia inhibition of aceticlastic methanogens instead of a non-competitive function. One batch experimental data set of anaerobic degradation of a raw waste activated sludge is used to calibrate the proposed model and three additional data sets from similar sludge thermally pretreated at three different temperatures are used to validate the parameters values.

  13. Effect of alkaline pretreatment on mesophilic and thermophilic anaerobic digestion of a submerged macrophyte: Inhibition and recovery against dissolved lignin during semi-continuous operation.

    PubMed

    Koyama, Mitsuhiko; Watanabe, Keiko; Kurosawa, Norio; Ishikawa, Kanako; Ban, Syuhei; Toda, Tatsuki

    2017-08-01

    The long-term effect of alkaline pretreatment on semi-continuous anaerobic digestion (AD) of the lignin-rich submerged macrophyte Potamogeton maackianus was investigated using mesophilic and thermophilic conditions. In pretreated reactors, dissolved lignin accumulated to high levels. CH 4 production under the pretreated condition was higher than that of the untreated condition, but decreased from Days 22 (mesophilic) and 42 (thermophilic). However, CH 4 production subsequently recovered, although dissolved lignin accumulated. Further, the change in the microbial community was observed between conditions. These results suggest that dissolved lignin temporarily inhibited AD, although acclimatization to dissolved lignin occurred during long-term operation. During the steady state period, mesophilic conditions achieved a 42% increase in the CH 4 yield using pretreatment, while thermophilic conditions yielded an 8% increment. Because volatile fatty acids accumulated even after acclimatization during the thermophilic pretreated condition and was discharged with the effluent, improvement of the methanogenic step would enable enhanced CH 4 recovery. Copyright © 2017 Elsevier Ltd. All rights reserved.

  14. Response surface optimization of substrates for thermophilic anaerobic codigestion of sewage sludge and food waste.

    PubMed

    Kim, Hyun-Woo; Shin, Hang-Sik; Han, Sun-Kee; Oh, Sae-Eun

    2007-03-01

    This study investigated the effects of food waste constituents on thermophilic (55 degrees C) anaerobic codigestion of sewage sludge and food waste by using statistical techniques based on biochemical methane potential tests. Various combinations of grain, vegetable, and meat as cosubstrate were tested, and then the data of methane potential (MP), methane production rate (MPR), and first-order kinetic constant of hydrolysis (kH) were collected for further analyses. Response surface methodology by the Box-Behnken design can verify the effects and their interactions of three variables on responses efficiently. MP was mainly affected by grain, whereas MPR and kH were affected by both vegetable and meat. Estimated polynomial regression models can properly explain the variability of experimental data with a high-adjusted R2 of 0.727, 0.836, and 0.915, respectively. By applying a series of optimization techniques, it was possible to find the proper criteria of cosubstrate. The optimal cosubstrate region was suggested based on overlay contours of overall mean responses. With the desirability contour plots, it was found that optimal conditions of cosubstrate for the maximum MPR (56.6 mL of CH4/g of chemical oxygen demand [COD]/day) were 0.71 g of COD/L of grain, 0.18 g of COD/L of vegetable, and 0.38 g of COD/L of meat by the simultaneous consideration of MP, MPR, and kH. Within the range of each factor examined, the corresponding optimal ratio of sewage sludge to cosubstrate was 71:29 as the COD basis. Elaborate discussions could yield practical operational strategies for the enhanced thermophilic anaerobic codigestion of sewage sludge and food waste.

  15. A new approach for concurrently improving performance of South Korean food waste valorization and renewable energy recovery via dry anaerobic digestion under mesophilic and thermophilic conditions.

    PubMed

    Nguyen, Dinh Duc; Yeop, Jeong Seong; Choi, Jaehoon; Kim, Sungsu; Chang, Soon Woong; Jeon, Byong-Hun; Guo, Wenshan; Ngo, Huu Hao

    2017-08-01

    Dry semicontinuous anaerobic digestion (AD) of South Korean food waste (FW) under four solid loading rates (SLRs) (2.30-9.21kg total solids (TS)/m 3 day) and at a fixed TS content was compared between two digesters, one each under mesophilic and thermophilic conditions. Biogas production and organic matter reduction in both digesters followed similar trends, increasing with rising SLR. Inhibitor (intermediate products of the anaerobic fermentation process) effects on the digesters' performance were not observed under the studied conditions. In all cases tested, the digesters' best performance was achieved at the SLR of 9.21kg TS/m 3 day, with 74.02% and 80.98% reduction of volatile solids (VS), 0.87 and 0.90m 3 biogas/kg VS removed , and 0.65 (65% CH 4 ) and 0.73 (60.02% CH 4 ) m 3 biogas/kg VS fed , under mesophilic and thermophilic conditions, respectively. Thermophilic dry AD is recommended for FW treatment in South Korea because it is more efficient and has higher energy recovery potential when compared to mesophilic dry AD. Copyright © 2017 Elsevier Ltd. All rights reserved.

  16. Comparison of the anaerobic digestion at the mesophilic and thermophilic temperature regime of organic wastes from the agribusiness.

    PubMed

    Almeida Streitwieser, Daniela

    2017-10-01

    An overall kinetic power law model has been successfully applied to study the anaerobic digestion of agricultural wastes. In this comparative kinetic study feed composition, organic load rate, residence time and process temperature have been systematically varied in an automated semi-continuous fermentation system to obtain the dependency of the rate of degradation as biogas production on the organic load rate and temperature. The results show that the overall reaction order depend only on the Chemical Oxygen Demand (COD) at values between 3.6 and 3.7. The Arrhenius approach shows a shift in the rate determining step between the mesophilic and thermophilic temperature regimes. The activation energy at the temperature insensitive mesophilic regime is very small at 8.9 (kJ/mole), while the activation energy at the temperature sensitive thermophilic regime lies around 117 (kJ/mole). Copyright © 2017 Elsevier Ltd. All rights reserved.

  17. An innovative intermittent-vacuum assisted thermophilic anaerobic digestion process for effective animal manure utilization and treatment.

    PubMed

    Zhang, Renchuan; Anderson, Erik; Addy, Min; Deng, Xiangyuan; Kabir, Fayal; Lu, Qian; Ma, Yiwei; Cheng, Yanling; Liu, Yuhuan; Chen, Paul; Ruan, Roger

    2017-11-01

    Intermittent-vacuum stripping (IVS) was developed as a pretreatment for thermophilic anaerobic digestion (TAD) to improve methanogenesis and hydrolysis activity through preventing free ammonia and hydrogen sulfide (H 2 S) inhibition from liquid swine manure (LSM). Over 98% of ammonia and 38% organic nitrogen were removed in 60min from 55°C to 85°C with vacuum pressure (from 100.63±3.79mmHg to 360.91±7.39mmHg) at initial pH 10.0 by IVS. Thermophilic methanogenesis and hydrolysis activity of pretreated LSM increased 52.25% (from 11.56±1.75% to 17.60±0.49%) in 25days and 40% (from 10days to 6days) in bio-methane potential assay. Over 80% H 2 S and total nitrogen were removed by IVS assistance, while around 70% nitrogen was recycled as ammonium sulfate. Therefore, IVS-TAD combination could be an effective strategy to improve TAD efficiency, whose elution is more easily utilized in algae cultivation and/or hydroponic system. Copyright © 2017 Elsevier Ltd. All rights reserved.

  18. [Isolation and identification of seven thermophilic and anaerobic bacteria from hot springs in Tengchong Rehai].

    PubMed

    Lu, Yueqing; Chen, Bo; Liu, Xiaoli; Ji, Xiuling; Wei, Yunlin; Lin, Lianbing

    2009-09-01

    In order to study the taxonomic characteristic and physiological, biochemical properties of anaerobic bacteria from hot springs in Tengchong Rehai, Yunnan Province, China. Using Hungate anaerobic technique We isolated seven strains from hot springs in Tengchong Rehai, Yunnan province, and analyzed their 16S rRNA gene sequences. The seven isolates were rod-shaped, Gram-negative, obligate anaerobe, and spores formation was not observed. All strains could grow well at 70 degrees C. Growth of strain RH0802 occurred between 60 and 80 degrees C, optimally around 70 degrees C. The pH range for its growth was between 5.5 and 8.5, with an optimum around 7.0. Strain RH0802 grew on a wide range of carbon sources, including glucose, starch, mannitol, mannose, ribose, maltose, cellobiose, xylose, fructose, galactose, xylan and glycerol, but it could not utilize sucrose or pyruvate. 16S rRNA gene phylogenetic analysis showed that the maximum similarity between the five strains and the strains of genus Caldanaerobacter was up to 98%, except RH0804 and RH0806, which reached to 96% and 93%, respectively. The two isolates were presumed to be potential novel species. The GenBank accession numbers of RH0802 to RH0808 were FJ748766, FJ748762, FJ748761, FJ748763, FJ748765, FJ748764 and FJ748767. The results showed that the seven thermophilic anaerobes belonged to the genus Caldanaerobacter.

  19. Influence of initial pH on thermophilic anaerobic co-digestion of swine manure and maize stalk.

    PubMed

    Zhang, Tong; Mao, Chunlan; Zhai, Ningning; Wang, Xiaojiao; Yang, Gaihe

    2015-01-01

    The contradictions between the increasing energy demand and decreasing fossil fuels are making the use of renewable energy the key to the sustainable development of energy in the future. Biogas, a renewable clean energy, can be obtained by the anaerobic fermentation of manure waste and agricultural straw. This study examined the initial pH value had obvious effect on methane production and the process in the thermophilic anaerobic co-digestion. Five different initial pH levels with three different manure ratios were tested. All digesters in different initial pH showed a diverse methane production after 35 days. The VFA/alkalinity ratio of the optimum reaction condition for methanogens activity was in the range of 0.1-0.3 and the optimal condition that at the 70% dung ratio and initial pH 6.81, was expected to achieve maximum total biogas production (146.32 mL/g VS). Copyright © 2014 Elsevier Ltd. All rights reserved.

  20. Quantitative Metaproteomics Highlight the Metabolic Contributions of Uncultured Phylotypes in a Thermophilic Anaerobic Digester

    PubMed Central

    Hagen, Live H.; Frank, Jeremy A.; Zamanzadeh, Mirzaman; Eijsink, Vincent G. H.; Pope, Phillip B.; Arntzen, Magnus Ø.

    2016-01-01

    ABSTRACT In this study, we used multiple meta-omic approaches to characterize the microbial community and the active metabolic pathways of a stable industrial biogas reactor with food waste as the dominant feedstock, operating at thermophilic temperatures (60°C) and elevated levels of free ammonia (367 mg/liter NH3-N). The microbial community was strongly dominated (76% of all 16S rRNA amplicon sequences) by populations closely related to the proteolytic bacterium Coprothermobacter proteolyticus. Multiple Coprothermobacter-affiliated strains were detected, introducing an additional level of complexity seldom explored in biogas studies. Genome reconstructions provided metabolic insight into the microbes that performed biomass deconstruction and fermentation, including the deeply branching phyla Dictyoglomi and Planctomycetes and the candidate phylum “Atribacteria.” These biomass degraders were complemented by a synergistic network of microorganisms that convert key fermentation intermediates (fatty acids) via syntrophic interactions with hydrogenotrophic methanogens to ultimately produce methane. Interpretation of the proteomics data also suggested activity of a Methanosaeta phylotype acclimatized to high ammonia levels. In particular, we report multiple novel phylotypes proposed as syntrophic acetate oxidizers, which also exert expression of enzymes needed for both the Wood-Ljungdahl pathway and β-oxidation of fatty acids to acetyl coenzyme A. Such an arrangement differs from known syntrophic oxidizing bacteria and presents an interesting hypothesis for future studies. Collectively, these findings provide increased insight into active metabolic roles of uncultured phylotypes and presents new synergistic relationships, both of which may contribute to the stability of the biogas reactor. IMPORTANCE Biogas production through anaerobic digestion of organic waste provides an attractive source of renewable energy and a sustainable waste management strategy. A

  1. Quantitative Metaproteomics Highlight the Metabolic Contributions of Uncultured Phylotypes in a Thermophilic Anaerobic Digester.

    PubMed

    Hagen, Live H; Frank, Jeremy A; Zamanzadeh, Mirzaman; Eijsink, Vincent G H; Pope, Phillip B; Horn, Svein J; Arntzen, Magnus Ø

    2017-01-15

    In this study, we used multiple meta-omic approaches to characterize the microbial community and the active metabolic pathways of a stable industrial biogas reactor with food waste as the dominant feedstock, operating at thermophilic temperatures (60°C) and elevated levels of free ammonia (367 mg/liter NH 3 -N). The microbial community was strongly dominated (76% of all 16S rRNA amplicon sequences) by populations closely related to the proteolytic bacterium Coprothermobacter proteolyticus. Multiple Coprothermobacter-affiliated strains were detected, introducing an additional level of complexity seldom explored in biogas studies. Genome reconstructions provided metabolic insight into the microbes that performed biomass deconstruction and fermentation, including the deeply branching phyla Dictyoglomi and Planctomycetes and the candidate phylum "Atribacteria" These biomass degraders were complemented by a synergistic network of microorganisms that convert key fermentation intermediates (fatty acids) via syntrophic interactions with hydrogenotrophic methanogens to ultimately produce methane. Interpretation of the proteomics data also suggested activity of a Methanosaeta phylotype acclimatized to high ammonia levels. In particular, we report multiple novel phylotypes proposed as syntrophic acetate oxidizers, which also exert expression of enzymes needed for both the Wood-Ljungdahl pathway and β-oxidation of fatty acids to acetyl coenzyme A. Such an arrangement differs from known syntrophic oxidizing bacteria and presents an interesting hypothesis for future studies. Collectively, these findings provide increased insight into active metabolic roles of uncultured phylotypes and presents new synergistic relationships, both of which may contribute to the stability of the biogas reactor. Biogas production through anaerobic digestion of organic waste provides an attractive source of renewable energy and a sustainable waste management strategy. A comprehensive understanding

  2. Molecular and conventional analyses of microbial diversity in mesophilic and thermophilic upflow anaerobic sludge blanket granular sludges.

    PubMed

    Sekiguchi, Y; Kamagata, Y; Ohashi, A; Harada, H

    2002-01-01

    The microbial community structure of mesophilic (35 degrees C) and thermophilic (55 degrees C) methanogenic granular sludges was surveyed by using both cultivation-independent molecular approach and conventional cultivation technique in order to address the fundamental questions on the microbial populations, i.e. who are present, where they are located, and what they are doing there. To elucidate the microbial constituents within both sludges, we first constructed 16S ribosomal DNA clone libraries, and partial sequencing of the clones was conducted for phylogenetic analysis. In this experiment, we found a number of unidentifiable clones within the domain Bacteria as well as clones that were closely related with 16S rDNAs of cultured microbes. The unidentifiable clones accounted for approximately 60-70% of the total clones in both mesophilic and thermophilic libraries. 16S rRNA-targeted in situ hybridization combined with confocal laser scanning microscopy was subsequently employed to examine where the uncultured populations were located within sludge granules. Spatial organization of uncultured microbes was visualized in thin-sections of both types of granules using fluorescent oligonucleotide probes, which were designed based on the clone sequences of certain novel clusters. This resulted in the detection of two types of uncultured cells in specific locations inside the granules. Finally, the goal-directed conventional cultivation technique was employed to recover such uncultured anaerobes and uncover their physiology and functions. In this approach, a total of five new species of thermophilic microorganisms were isolated, including several types of syntrophs and a novel sugar-fermenting bacterium. In the previous molecular approaches, all of these isolates were suggested to be significant populations within thermophilic granular sludge, hence obtaining these isolates in pure culture decreased the fraction of unknown clones in the previous thermophilic clone

  3. Kinetics of butyrate, acetate, and hydrogen metabolism in a thermophilic, anaerobic, butyrate-degrading triculture.

    PubMed

    Ahring, B K; Westermann, P

    1987-02-01

    Kinetics of butyrate, acetate, and hydrogen metabolism were determined with butyrate-limited, chemostat-grown tricultures of a thermophilic butyrate-utilizing bacterium together with Methanobacterium thermoautotrophicum and the TAM organism, a thermophilic acetate-utilizing methanogenic rod. Kinetic parameters were determined from progress curves fitted to the integrated form of the Michaelis-Menten equation. The apparent half-saturation constants, K(m), for butyrate, acetate, and dissolved hydrogen were 76 muM, 0.4 mM, and 8.5 muM, respectively. Butyrate and hydrogen were metabolized to a concentration of less than 1 muM, whereas acetate uptake usually ceased at a concentration of 25 to 75 muM, indicating a threshold level for acetate uptake. No significant differences in K(m) values for butyrate degradation were found between chemostat- and batch-grown tricultures, although the maximum growth rate was somewhat higher in the batch cultures in which the medium was supplemented with yeast extract. Acetate utilization was found to be the rate-limiting reaction for complete degradation of butyrate to methane and carbon dioxide in continuous culture. Increasing the dilution rate resulted in a gradual accumulation of acetate. The results explain the low concentrations of butyrate and hydrogen normally found during anaerobic digestion and the observation that acetate is the first volatile fatty acid to accumulate upon a decrease in retention time or increase in organic loading of a digestor.

  4. Enhancing the electron transfer capacity and subsequent color removal in bioreactors by applying thermophilic anaerobic treatment and redox mediators.

    PubMed

    dos Santos, A B; Traverse, J; Cervantes, F J; van Lier, J B

    2005-01-05

    The effect of temperature, hydraulic retention time (HRT) and the redox mediator anthraquinone-2,6-disulfonate (AQDS), on electron transfer and subsequent color removal from textile wastewater was assessed in mesophilic and thermophilic anaerobic bioreactors. The results clearly show that compared with mesophilic anaerobic treatment, thermophilic treatment at 55 degrees C is an effective approach for increasing the electron transfer capacity in bioreactors, and thus improving the decolorization rates. Furthermore, similar color removals were found at 55 degrees C between the AQDS-free and AQDS-supplemented reactors, whereas a significant difference (up to 3.6-fold) on decolorization rates occurred at 30 degrees C. For instance, at an HRT of 2.5 h and in the absence of AQDS, the color removal was 5.3-fold higher at 55 degrees C compared with 30 degrees C. The impact of a mix of mediators with different redox potentials on the decolorization rate was investigated with both industrial textile wastewater and the azo dye Reactive Red 2 (RR2). Color removal of RR2 in the presence of anthraquinone-2-sulfonate (AQS) (standard redox potential E(0)' of -225 mV) was 3.8-fold and 2.3-fold higher at 30 degrees C and 55 degrees C, respectively, than the values found in the absence of AQS. Furthermore, when the mediators 1,4-benzoquinone (BQ) (E(0)' of +280 mV), and AQS were incubated together, there was no improvement on the decolorization rates compared with the bottles solely supplemented with AQS. Results imply that the use of mixed redox mediators with positive and negative E(0)' under anaerobic conditions is not an efficient approach to improve color removal in textile wastewaters. (c) 2004 Wiley Periodicals, Inc.

  5. Development of an empirical mathematical model for describing and optimizing the hygiene potential of a thermophilic anaerobic bioreactor treating faeces.

    PubMed

    Lübken, M; Wichern, M; Bischof, F; Prechtl, S; Horn, H

    2007-01-01

    Poor sanitation and insufficient disposal of sewage and faeces are primarily responsible for water associated health problems in developing countries. Domestic sewage and faeces are prevalently discharged into surface waters which are used by the inhabitants as a source for drinking water. This paper presents a decentralized anaerobic process technique for handling of such domestic organic waste. Such an efficient and compact system for treating faeces and food waste may be of great benefit for developing countries. Besides a stable biogas production for energy generation, the reduction of bacterial pathogens is of particular importance. In our research we investigated the removal capacity of the reactor concerning pathogens, which has been operated under thermophilic conditions. Faecal coliforms and intestinal enterococci have been detected as indicator organisms for bacterial pathogens. By the multiple regression analysis technique an empirical mathematical model has been developed. The model shows a high correlation between removal efficiency and both, hydraulic retention time (HRT) and temperature. By this model an optimized HRT for defined bacterial pathogens effluent standards can be easily calculated. Thus, hygiene potential can be evaluated along with economic aspects. In this paper not only results for describing the hygiene potential of a thermophilic anaerobic bioreactor are presented, but also an exemplary method to draw the right conclusions out of biological tests with the aid of mathematical tools.

  6. Analysis of methanogenic activity in a thermophilic-dry anaerobic reactor: use of fluorescent in situ hybridization.

    PubMed

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

    2009-03-01

    Methanogenic activity in a thermophilic-dry anaerobic reactor was determined by comparing the amount of methane generated for each of the organic loading rates with the size of the total and specific methanogenic population, as determined by fluorescent in situ hybridization. A high correlation was evident between the total methanogenic activity and retention time [-0.6988Ln(x)+2.667] (R(2) 0.8866). The total methanogenic activity increased from 0.04x10(-8) mLCH(4) cell(-1)day(-1) to 0.38x10(-8) mLCH(4) cell(-1)day(-1) while the retention time decreased, augmenting the organic loading rates. The specific methanogenic activities of H(2)-utilizing methanogens and acetate-utilizing methanogens increased until they stabilised at 0.64x10(-8) mLCH(4) cell(-1)day(-1) and 0.33x10(-8) mLCH(4) cell(-1)day(-1), respectively. The methanogenic activity of H(2)-utilizing methanogens was higher than acetate-utilizing methanogens, indicating that maintaining a low partial pressure of hydrogen does not inhibit the acetoclastic methanogenesis or the anaerobic process.

  7. Drivers of microbial community composition in mesophilic and thermophilic temperature-phased anaerobic digestion pre-treatment reactors.

    PubMed

    Pervin, Hasina M; Dennis, Paul G; Lim, Hui J; Tyson, Gene W; Batstone, Damien J; Bond, Philip L

    2013-12-01

    Temperature-phased anaerobic digestion (TPAD) is an emerging technology that facilitates improved performance and pathogen destruction in anaerobic sewage sludge digestion by optimising conditions for 1) hydrolytic and acidogenic organisms in a first-stage/pre-treatment reactor and then 2) methogenic populations in a second stage reactor. Pre-treatment reactors are typically operated at 55-65 °C and as such select for thermophilic bacterial communities. However, details of key microbial populations in hydrolytic communities and links to functionality are very limited. In this study, experimental thermophilic pre-treatment (TP) and control mesophilic pre-treatment (MP) reactors were operated as first-stages of TPAD systems treating activated sludge for 340 days. The TP system was operated sequentially at 50, 60 and 65 °C, while the MP rector was held at 35 °C for the entire period. The composition of microbial communities associated with the MP and TP pre-treatment reactors was characterised weekly using terminal-restriction fragment length polymorphism (T-RFLP) supported by clone library sequencing of 16S rRNA gene amplicons. The outcomes of this approach were confirmed using 454 pyrosequencing of gene amplicons and fluorescence in-situ hybridisation (FISH). TP associated bacterial communities were dominated by populations affiliated to the Firmicutes, Thermotogae, Proteobacteria and Chloroflexi. In particular there was a progression from Thermotogae to Lutispora and Coprothermobacter and diversity decreased as temperature and hydrolysis performance increased. While change in the composition of TP associated bacterial communities was attributable to temperature, that of MP associated bacterial communities was related to the composition of the incoming feed. This study determined processes driving the dynamics of key microbial populations that are correlated with an enhanced hydrolytic functionality of the TPAD system. Copyright © 2013 Elsevier Ltd. All rights

  8. Growing Chlorella vulgaris on thermophilic anaerobic digestion swine manure for nutrient removal and biomass production.

    PubMed

    Deng, Xiang-Yuan; Gao, Kun; Zhang, Ren-Chuan; Addy, Min; Lu, Qian; Ren, Hong-Yan; Chen, Paul; Liu, Yu-Huan; Ruan, Roger

    2017-11-01

    Liquid swine manure was subjected to thermophilic anaerobic digestion, ammonia stripping and centrifugation in order to increase the available carbon sources and decrease the ammonia concentration and turbidity. Chlorella vulgaris (UTEX 2714) was grown on minimally diluted (2×, 3× and 4×) autoclaved and non-autoclaved pretreated anaerobic digestion swine manure (PADSM) in a batch-culture system for 7days. Results showed that C. vulgaris (UTEX 2714) grew best on 3× PADSM media, and effectively removed NH 4 + -N, TN, TP and COD by 98.5-99.8%, 49.2-55.4%, 20.0-29.7%, 31.2-34.0% and 99.8-99.9%, 67.4-70.8%, 49.3-54.4%, 73.6-78.7% in differently diluted autoclaved and non-autoclaved PADSM, respectively. Results of chemical compositions indicated that contents of pigment, carbohydrate, protein and lipid in C. vulgaris (UTEX 2714) changed with the culture conditions. Moreover, its fatty acid profiles suggested that this alga could be used as animal feed if cultivated in autoclaved PADSM or as good-quality biodiesel feedstock if cultivated in non-autoclaved PADSM. Copyright © 2017 Elsevier Ltd. All rights reserved.

  9. Mechanism and Effect of Temperature on Variations in Antibiotic Resistance Genes during Anaerobic Digestion of Dairy Manure.

    PubMed

    Sun, Wei; Qian, Xun; Gu, Jie; Wang, Xiao-Juan; Duan, Man-Li

    2016-07-22

    Animal manure comprises an important reservoir for antibiotic resistance genes (ARGs), but the variation in ARGs during anaerobic digestion at various temperatures and its underlying mechanism remain unclear. Thus, we performed anaerobic digestion using dairy manure at three temperature levels (moderate: 20 °C, mesophilic: 35 °C, and thermophilic: 55 °C), to analyze the dynamics of ARGs and bacterial communities by quantitative PCR and 16S rRNA gene sequencing. We found that 8/10 detected ARGs declined and 5/10 decreased more than 1.0 log during thermophilic digestion, whereas only four and five ARGs decreased during moderate and mesophilic digestion, respectively. The changes in ARGs and bacterial communities were similar under the moderate and mesophilic treatments, but distinct from those in the thermophilic system. Potential pathogens such as Bacteroidetes, Proteobacteria, and Corynebacterium were removed by thermophilic digestion but not by moderate and mesophilic digestion. The bacterial community succession was the dominant mechanism that influenced the variation in ARGs and integrons during anaerobic digestion. Thermophilic digestion decreased the amount of mesophilic bacteria (Bacteroidetes and Proteobacteria) carrying ARGs. Anaerobic digestion generally decreased the abundance of integrons by eliminating the aerobic hosts of integrons (Actinomycetales and Bacilli). Thermophilic anaerobic digestion is recommended for the treatment and reuse of animal manure.

  10. Mechanism and Effect of Temperature on Variations in Antibiotic Resistance Genes during Anaerobic Digestion of Dairy Manure

    NASA Astrophysics Data System (ADS)

    Sun, Wei; Qian, Xun; Gu, Jie; Wang, Xiao-Juan; Duan, Man-Li

    2016-07-01

    Animal manure comprises an important reservoir for antibiotic resistance genes (ARGs), but the variation in ARGs during anaerobic digestion at various temperatures and its underlying mechanism remain unclear. Thus, we performed anaerobic digestion using dairy manure at three temperature levels (moderate: 20 °C, mesophilic: 35 °C, and thermophilic: 55 °C), to analyze the dynamics of ARGs and bacterial communities by quantitative PCR and 16S rRNA gene sequencing. We found that 8/10 detected ARGs declined and 5/10 decreased more than 1.0 log during thermophilic digestion, whereas only four and five ARGs decreased during moderate and mesophilic digestion, respectively. The changes in ARGs and bacterial communities were similar under the moderate and mesophilic treatments, but distinct from those in the thermophilic system. Potential pathogens such as Bacteroidetes, Proteobacteria, and Corynebacterium were removed by thermophilic digestion but not by moderate and mesophilic digestion. The bacterial community succession was the dominant mechanism that influenced the variation in ARGs and integrons during anaerobic digestion. Thermophilic digestion decreased the amount of mesophilic bacteria (Bacteroidetes and Proteobacteria) carrying ARGs. Anaerobic digestion generally decreased the abundance of integrons by eliminating the aerobic hosts of integrons (Actinomycetales and Bacilli). Thermophilic anaerobic digestion is recommended for the treatment and reuse of animal manure.

  11. Mesophilic and thermophilic anaerobic co-digestion of winery wastewater sludge and wine lees: An integrated approach for sustainable wine production.

    PubMed

    Da Ros, C; Cavinato, C; Pavan, P; Bolzonella, D

    2017-12-01

    In this work, winery wastes generated by a cellar producing approximately 300,000 hL of wine per year was monitored for a period of one year. On average, 196 L of wastewater, 0.1 kg of waste activated sludge (dry matter) and 1.6 kg of wine lees were produced per hectoliter of wine produced. Different winery wastes, deriving from different production steps, namely waste activated sludge from wastewater treatment and wine lees, were co-treated using an anaerobic digestion process. Testing was conducted on a pilot scale for both mesophilic and thermophilic conditions. The process was stable for a long period at 37 °C, with an average biogas production of 0.386 m 3 /kg COD fed . On the other hand, for thermophilic conditions, volatile fatty acids accumulated in the reactor and the process failed after one hydraulic retention time (23 days). In order to fix the biological process, trace elements (iron, cobalt and nickel) were added to the feed of the thermophilic reactor. Metals augmentation improved process stability and yields at 55 °C. The pH ranged between 7.8 and 8.0, and specific gas production was 0.450 m 3 /kg COD fed , which corresponded to dry matter and COD removals of 34% and 88%, respectively. Although the observed performances in terms of biogas production were good, the thermophilic process exhibited some limitations related to both the necessity of metals addition and the worse dewaterability properties. In fact, while the mesophilic digestates reached a good dewatering quality via the addition of 6.5 g of polymer per kg of dry matter, the required dosage for the thermophilic sludge was greater than 10 g/kg of dry matter. Copyright © 2016 Elsevier Ltd. All rights reserved.

  12. A comparison of microbial characteristics between the thermophilic and mesophilic anaerobic digesters exposed to elevated food waste loadings.

    PubMed

    Guo, Xiaohui; Wang, Cheng; Sun, Faqian; Zhu, Weijing; Wu, Weixiang

    2014-01-01

    Thermophilic and mesophilic anaerobic digestion reactors (TR and MR) using food waste as substrate were compared with emphasis on microbial responses to increasing organic loading rate (OLR). At OLR ranging from 1.0 to 2.5 g VS L(-1) d(-1), MR exhibited more stable performance compared to TR in terms of methane yield. Amplicons pyrosequencing results revealed the distinct microbial dynamics in the two reactors. Primarily, MR had greater richness and evenness of bacteria species. With OLR elevated, larger shifts of bacterial phylogeny were observed in MR; Methanosaeta dominated in archaeal community in MR while Methanothermobacter and Methanoculleus were favored in TR. The high functional redundancy in bacterial community integrated with acetoclastic methanogenesis in MR resulted in its better performance; whereas delicate interactions between hydrogen-producer and hydrogenotrophic methanogens in TR were much more prone to disruption. These results are conductive to understanding the microbial mechanisms of low methane yield during food waste anaerobic digestion. Copyright © 2013 Elsevier Ltd. All rights reserved.

  13. Mechanism and Effect of Temperature on Variations in Antibiotic Resistance Genes during Anaerobic Digestion of Dairy Manure

    PubMed Central

    Sun, Wei; Qian, Xun; Gu, Jie; Wang, Xiao-Juan; Duan, Man-Li

    2016-01-01

    Animal manure comprises an important reservoir for antibiotic resistance genes (ARGs), but the variation in ARGs during anaerobic digestion at various temperatures and its underlying mechanism remain unclear. Thus, we performed anaerobic digestion using dairy manure at three temperature levels (moderate: 20 °C, mesophilic: 35 °C, and thermophilic: 55 °C), to analyze the dynamics of ARGs and bacterial communities by quantitative PCR and 16S rRNA gene sequencing. We found that 8/10 detected ARGs declined and 5/10 decreased more than 1.0 log during thermophilic digestion, whereas only four and five ARGs decreased during moderate and mesophilic digestion, respectively. The changes in ARGs and bacterial communities were similar under the moderate and mesophilic treatments, but distinct from those in the thermophilic system. Potential pathogens such as Bacteroidetes, Proteobacteria, and Corynebacterium were removed by thermophilic digestion but not by moderate and mesophilic digestion. The bacterial community succession was the dominant mechanism that influenced the variation in ARGs and integrons during anaerobic digestion. Thermophilic digestion decreased the amount of mesophilic bacteria (Bacteroidetes and Proteobacteria) carrying ARGs. Anaerobic digestion generally decreased the abundance of integrons by eliminating the aerobic hosts of integrons (Actinomycetales and Bacilli). Thermophilic anaerobic digestion is recommended for the treatment and reuse of animal manure. PMID:27444518

  14. A comparative study on the alternating mesophilic and thermophilic two-stage anaerobic digestion of food waste.

    PubMed

    Ventura, Jey-R Sabado; Lee, Jehoon; Jahng, Deokjin

    2014-06-01

    An alternating mesophilic and thermophilic two stage anaerobic digestion (AD) process was conducted. The temperature of the acidogenic (A) and methanogenic (M) reactors was controlled as follows: System 1 (S1) mesophilic A-mesophilic M; (S2) mesophilic A-thermophilic M; and (S3) thermophilic A-mesophilic M. Initially, the AD reactor was acclimatized and inoculated with digester sludge. Food waste was added with the soluble chemical oxygen demand (SCOD) concentrations of 41.4-47.0 g/L and volatile fatty acids of 2.0-3.2 g/L. Based on the results, the highest total chemical oxygen demand removal (86.6%) was recorded in S2 while S3 exhibited the highest SCOD removal (96.6%). Comparing S1 with S2, total solids removal increased by 0.5%; S3 on the other hand decreased by 0.1 % as compared to S1. However, volatile solids (VS) removal in S1, S2, and S3 was 78.5%, 81.7%, and 79.2%, respectively. S2 also exhibited the highest CH4 content, yield, and production rate of 70.7%, 0.44 L CH4/g VSadded, and 1.23 L CH4/(L·day), respectively. Bacterial community structure revealed that the richness, diversity, evenness, and dominance of S2 were high except for the archaeal community. The terminal restriction fragments dendrogram also revealed that the microbial community of the acidogenic and methanogenic reactors in S2 was distinct. Therefore, S2 was the best among the systems for the operation of two-stage AD of food waste in terms of CH4 production, nutrient removal, and microbial community structure. Copyright © 2014 The Research Centre for Eco-Environmental Sciences, Chinese Academy of Sciences. Published by Elsevier B.V. All rights reserved.

  15. A Combination of Stable Isotope Probing, Illumina Sequencing, and Co-occurrence Network to Investigate Thermophilic Acetate- and Lactate-Utilizing Bacteria.

    PubMed

    Sun, Weimin; Krumins, Valdis; Dong, Yiran; Gao, Pin; Ma, Chunyan; Hu, Min; Li, Baoqin; Xia, Bingqing; He, Zijun; Xiong, Shangling

    2018-01-01

    Anaerobic digestion is a complicated microbiological process that involves a wide diversity of microorganisms. Acetate is one of the most important intermediates, and interactions between acetate-oxidizing bacteria and archaea could play an important role in the formation of methane in anoxic environments. Anaerobic digestion at thermophilic temperatures is known to increase methane production, but the effects on the microbial community are largely unknown. In the current study, stable isotope probing was used to characterize acetate- and lactate-oxidizing bacteria in thermophilic anaerobic digestion. In microcosms fed 13 C-acetate, bacteria related to members of Clostridium, Hydrogenophaga, Fervidobacterium, Spirochaeta, Limnohabitans, and Rhodococcus demonstrated elevated abundances of 13 C-DNA fractions, suggesting their activities in acetate oxidation. In the treatments fed 13 C-lactate, Anaeromyxobacter, Desulfobulbus, Syntrophus, Cystobacterineae, and Azospira were found to be the potential thermophilic lactate utilizers. PICRUSt predicted that enzymes related to nitrate and nitrite reduction would be enriched in 13 C-DNA fractions, suggesting that the acetate and lactate oxidation may be coupled with nitrate and/or nitrite reduction. Co-occurrence network analysis indicated bacterial taxa not enriched in 13 C-DNA fractions that may also play a critical role in thermophilic anaerobic digestion.

  16. Simultaneous hydrolysis and fermentation of unprocessed food waste into ethanol using thermophilic anaerobic bacteria.

    PubMed

    Dhiman, Saurabh Sudha; David, Aditi; Shrestha, Namita; Johnson, Glenn R; Benjamin, Kenneth M; Gadhamshetty, Venkataramana; Sani, Rajesh K

    2017-11-01

    The one-pot CRUDE (Conversion of Raw and Untreated Disposal into Ethanol) process was developed for simultaneous hydrolysis and fermentation of unprocessed food waste into ethanol using thermophilic (growing at 65°C) anaerobic bacteria. Unlike existing waste to energy technologies, the CRUDE process obviates the need for any pre-treatment or enzyme addition. A High-Temperature-High-Pressure (HTHP) distillation technique was also applied that facilitated efficient use of fermentation medium, inoculum recycling, and in-situ ethanol collection. For material balancing of the process, each characterized component was represented in terms of C-mol. Recovery of 94% carbon at the end confirmed the operational efficiency of CRUDE process. The overall energy retaining efficiency calculated from sugars to ethanol was 1262.7kJdryweightkg -1 of volatile solids using HTHP. These results suggest that the CRUDE process can be a starting point for the development of a commercial ethanol production process. Copyright © 2017 Elsevier Ltd. All rights reserved.

  17. Ethanol and anaerobic conditions reversibly inhibit commercial cellulase activity in thermophilic simultaneous saccharification and fermentation (tSSF)

    PubMed Central

    2012-01-01

    Background A previously developed mathematical model of low solids thermophilic simultaneous saccharification and fermentation (tSSF) with Avicel was unable to predict performance at high solids using a commercial cellulase preparation (Spezyme CP) and the high ethanol yield Thermoanaerobacterium saccharolyticum strain ALK2. The observed hydrolysis proceeded more slowly than predicted at solids concentrations greater than 50 g/L Avicel. Factors responsible for this inaccuracy were investigated in this study. Results Ethanol dramatically reduced cellulase activity in tSSF. At an Avicel concentration of 20 g/L, the addition of ethanol decreased conversion at 96 hours, from 75% in the absence of added ethanol down to 32% with the addition of 34 g/L initial ethanol. This decrease is much greater than expected based on hydrolysis inhibition results in the absence of a fermenting organism. The enhanced effects of ethanol were attributed to the reduced, anaerobic conditions of tSSF, which were shown to inhibit cellulase activity relative to hydrolysis under aerobic conditions. Cellulose hydrolysis in anaerobic conditions was roughly 30% slower than in the presence of air. However, this anaerobic inhibition was reversed by exposing the cellulase enzymes to air. Conclusion This work demonstrates a previously unrecognized incompatibility of enzymes secreted by an aerobic fungus with the fermentation conditions of an anaerobic bacterium and suggests that enzymes better suited to industrially relevant fermentation conditions would be valuable. The effects observed may be due to inactivation or starvation of oxygen dependent GH61 activity, and manipulation or replacement of this activity may provide an opportunity to improve biomass to fuel process efficiency. PMID:22703989

  18. Methanobacterium thermoautotrophicus sp. n., an anaerobic, autotrophic, extreme thermophile.

    PubMed

    Zeikus, J G; Wolfe, R S

    1972-02-01

    The isolation of a new methanogenic bacterium, Methanobacterium thermoautotrophicus sp. n., is described. Successful isolation required a medium containing inorganic salts, an atmosphere consisting of an 80:20 mixture of hydrogen-carbon dioxide, and incubation temperatures of 65 to 70 C. Isolates of M. thermoautotrophicus were gram-positive, nonmotile, irregularly curved rods which frequently formed long filaments. The organism was found to be an autotroph and a strict anaerobe, and to have a pH optimum of 7.2 to 7.6. The optimal temperature for growth was 65 to 70 C, the maximum being 75 C and the minimum about 40 C. The generation time at the optimum was about 5 hr. The deoxyribonucleic acid of M. thermoautotrophicus had a guanine plus cytosine (GC) content of 52 moles per cent, whereas Methanobacterium sp. strain M.O.H. had a GC content of 38%. When heated, intact ribosomes of Methanobacterium sp. strain M.O.H. were stable up to 55 C and had a T(m) of 73 C. In contrast, ribosomes of M. thermoautotrophicus were stable up to 75 C and had a T(m) of 82 C. Upon complete thermal denaturation, ribosomes of strain M.O.H. underwent a 59% hyperchromic shift, whereas those of the thermophile showed only a 20% increase in hyperchromicity. Methane formation in cell-free extracts of M. thermoautotrophicus was temperature-dependent and required hydrogen and carbon dioxide; methyl cobalamin served as a methyl donor, and addition of coenzyme M stimulated methanogenesis.

  19. Methanobacterium thermoautotrophicus sp. n., an Anaerobic, Autotrophic, Extreme Thermophile

    PubMed Central

    Zeikus, J. G.; Wolee, R. S.

    1972-01-01

    The isolation of a new methanogenic bacterium, Methanobacterium thermoautotrophicus sp. n., is described. Successful isolation required a medium containing inorganic salts, an atmosphere consisting of an 80:20 mixture of hydrogen-carbon dioxide, and incubation temperatures of 65 to 70 C. Isolates of M. thermoautotrophicus were gram-positive, nonmotile, irregularly curved rods which frequently formed long filaments. The organism was found to be an autotroph and a strict anaerobe, and to have a pH optimum of 7.2 to 7.6. The optimal temperature for growth was 65 to 70 C, the maximum being 75 C and the minimum about 40 C. The generation time at the optimum was about 5 hr. The deoxyribonucleic acid of M. thermoautotrophicus had a guanine plus cytosine (GC) content of 52 moles per cent, whereas Methanobacterium sp. strain M.O.H. had a GC content of 38%. When heated, intact ribosomes of Methanobacterium sp. strain M.O.H. were stable up to 55 C and had a Tm of 73 C. In contrast, ribosomes of M. thermoautotrophicus were stable up to 75 C and had a Tm of 82 C. Upon complete thermal denaturation, ribosomes of strain M.O.H. underwent a 59% hyperchromic shift, whereas those of the thermophile showed only a 20% increase in hyperchromicity. Methane formation in cell-free extracts of M. thermoautotrophicus was temperature-dependent and required hydrogen and carbon dioxide; methyl cobalamin served as a methyl donor, and addition of coenzyme M stimulated methanogenesis. Images PMID:4550816

  20. Moorella stamsii sp. nov., a new anaerobic thermophilic hydrogenogenic carboxydotroph isolated from digester sludge.

    PubMed

    Alves, J I; van Gelder, A H; Alves, M M; Sousa, D Z; Plugge, C M

    2013-11-01

    A novel anaerobic, thermophilic, carbon monoxide-utilizing bacterium, strain E3-O(T), was isolated from anaerobic sludge from a municipal solid waste digester. Cells were straight rods, 0.6-1 µm in diameter and 2-3 µm in length and grew as single cells or in pairs. Cells formed round terminal endospores. The temperature range for growth was 50-70 °C, with an optimum at 65 °C. The pH range for growth was 5.7-8.0, with an optimum at 7.5. Strain E3-O(T) had the ability to ferment various sugars, such as fructose, galactose, glucose, mannose, raffinose, ribose, sucrose and xylose, producing mainly H2 and acetate. In addition, the isolate was able to grow with CO as the sole carbon and energy source. CO oxidation was coupled to H2 and CO2 formation. The G+C content of the genomic DNA was 54.6 mol%. Based on 16S rRNA gene sequence analysis, this bacterium is most closely related to Moorella glycerini (97 % sequence identity). Based on the physiological features and phylogenetic analysis, it is proposed that strain E3-O(T) should be classified in the genus Moorella as a representative of a novel species, Moorella stamsii. The type strain of Moorella stamsii is E3-O(T) ( = DSM 26271(T) = CGMCC 1.5181(T)).

  1. Performance and kinetic study of semi-dry thermophilic anaerobic digestion of organic fraction of municipal solid waste

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Sajeena Beevi, B., E-mail: sajeenanazer@gmail.com; Madhu, G., E-mail: profmadhugopal@gmail.com; Sahoo, Deepak Kumar, E-mail: dksahoo@gmail.com

    2015-02-15

    Highlights: • Performance of the reactor was evaluated by the degradation of volatile solids. • Biogas yield at the end of the digestion was 52.9 L/kg VS. • Value of reaction rate constant, k, obtained was 0.0249 day{sup −1}. • During the digestion 66.7% of the volatile solid degradation was obtained. - Abstract: Anaerobic digestion (AD) of the organic fraction of municipal solid waste (OFMSW) is promoted as an energy source and waste disposal. In this study semi dry anaerobic digestion of organic solid wastes was conducted for 45 days in a lab-scale batch experiment for total solid concentration ofmore » 100 g/L for investigating the start-up performances under thermophilic condition (50 °C). The performance of the reactor was evaluated by measuring the daily biogas production and calculating the degradation of total solids and the total volatile solids. The biogas yield at the end of the digestion was 52.9 L/kg VS (volatile solid) for the total solid (TS) concentration of 100 g/L. About 66.7% of the volatile solid degradation was obtained during the digestion. A first order model based on the availability of substrate as the limiting factor was used to perform the kinetic studies of batch anaerobic digestion system. The value of reaction rate constant, k, obtained was 0.0249 day{sup −1}.« less

  2. Variation of the microbial community in thermophilic anaerobic digestion of pig manure mixed with different ratios of rice straw.

    PubMed

    Zhou, Sheng; Nikolausz, Marcell; Zhang, Jining; Riya, Shohei; Terada, Akihiko; Hosomi, Masaaki

    2016-09-01

    The effect of pig manure mixed with rice straw on methane yield and the microbial community involved in a thermophilic (55°C) anaerobic digestion process was investigated. Three substrates composed of mixed pig manure and rice straw at different ratios (95:5; 78:22 and 65:35 w/w, which resulted in C/N ratios of 10:1, 20:1 and 30:1) were used for the experiment. The substrate type had a major influence on the total bacterial community, while the methanogens were less affected. The members of the class Clostridia (phylum Firmicutes) were predominant regardless of mixture ratio (C/N ratio), but at species level there was a major difference between the low and high C/N ratio samples. The hydrogenotrophic methanogenic genus of Methanothermobacter was predominant in all samples but higher C/N ratio sequences affiliated to the genus Methanosarcina were also detected. The appearance of Methanosarcina sp. is most likely due to the less inhibition of ammonia during the anaerobic digestion. Copyright © 2016 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.

  3. Sequential batch thermophilic solid-state anaerobic digestion of lignocellulosic biomass via recirculating digestate as inoculum - Part II: Microbial diversity and succession.

    PubMed

    Lin, Long; Yu, Zhongtang; Li, Yebo

    2017-10-01

    This study aimed to investigate the effect of recirculation of digestate as inoculum on the microbial communities in thermophilic solid-state anaerobic digestion (SS-AD) of yard trimmings. The SS-AD consisted of 4 consecutive runs (30days/run), with digestate from the previous run being used as the inoculum of the subsequent run. Bacterial and archaeal communities (day 0, 4, 8, 12, 20, and 30) were examined using Illumina sequencing of 16S rRNA genes. The results revealed substantial microbial succession toward increased diversity until run 3. The proportions of Firmicutes that contained cellulolytic bacteria doubled, which might explain the concomitantly increased cellulose degradation and volatile fatty acids (VFAs). Clostridia and Thermotogae appeared to correlate with VFAs. The VFA accumulation likely induced dynamic shifts of methanogens, particularly to hydrogenotrophic Methanothermobacter, implying that non-acetoclastic oxidative pathway dominated during the steady-state thermophilic SS-AD. This study suggested that recirculating SS-AD digestate might be an effective way for inoculation. Copyright © 2017 Elsevier Ltd. All rights reserved.

  4. Economic Feasibility of Installing an Anaerobic Digester on a Department of Defense Installation

    DTIC Science & Technology

    2010-03-01

    permits anaerobic bacteria and enzymes to affect more waste than a lagoon does, as well as preventing a film or layer of scum forming on top of the waste...temperature classifications for anaerobic digestion. The three classes listed are: psychrophilic (4- 20 C), mesophilic (20-45 C), and thermophilic (45-60...operated at 55º C, 30 focusing on an optimum temperature for thermophilic bacteria. Despite previously discussed percentages for total solids in

  5. Use of respirometer in evaluation of process and toxicity of thermophilic anaerobic digestion for treating kitchen waste.

    PubMed

    Kuo, Wen-Chien; Cheng, Kae-Yiin

    2007-07-01

    A thermophilic anaerobic digestion (TAnD, 55 degrees C) system was adopted to hydrolyze the kitchen waste for 3 days, which was then fermented for a hydraulic retention time (HRT) of 10 days. The TAnD system performed much better than a similar system without thermal pre-treatment. A bubble respirometer was employed to study the effects of thermal pre-treatment, which showed that pre-treatment at 60 degrees C yielded the highest Total COD (TCOD) removal efficiency (79.2%) after 300h reaction. Respirometer results also indicated that oil and grease (O and G) began to inhibit the TAnD system at a concentration of approximately 1000mg/L and the gas production was inhibited by 50% at a concentration of approximately 7500mg/L of sodium.

  6. Anaerobic co-digestion of dairy manure and potato waste

    NASA Astrophysics Data System (ADS)

    Yadanaparthi, Sai Krishna Reddy

    Dairy and potato are two important agricultural commodities in Idaho. Both the dairy and potato processing industries produce a huge amount of waste which could cause environmental pollution. To minimize the impact of potential pollution associated with dairy manure (DM) and potato waste (PW), anaerobic co-digestion has been considered as one of the best treatment process. The purpose of this research is to evaluate the anaerobic co-digestion of dairy manure and potato waste in terms of process stability, biogas generation, construction and operating costs, and potential revenue. For this purpose, I conducted 1) a literature review, 2) a lab study on anaerobic co-digestion of dairy manure and potato waste at three different temperature ranges (ambient (20-25°C), mesophilic (35-37°C) and thermophilic (55-57°C) with five mixing ratios (DM:PW-100:0, 90:10, 80:20, 60:40, 40:60), and 3) a financial analysis for anaerobic digesters based on assumed different capital costs and the results from the lab co-digestion study. The literature review indicates that several types of organic waste were co-digested with DM. Dairy manure is a suitable base matter for the co-digestion process in terms of digestion process stability and methane (CH4) production (Chapter 2). The lab tests showed that co-digestion of DM with PW was better than digestion of DM alone in terms of biogas and CH4 productions (Chapter 3). The financial analysis reveals DM and PW can be used as substrate for full size anaerobic digesters to generate positive cash flow within a ten year time period. Based on this research, the following conclusions and recommendations were made: ▸ The ratio of DM:PW-80:20 is recommended at thermophilic temperatures and the ratio of DM:PW-90:10 was recommended at mesophilic temperatures for optimum biogas and CH4 productions. ▸ In cases of anaerobic digesters operated with electricity generation equipment (generators), low cost plug flow digesters (capital cost of 600/cow

  7. Comparison of multi-enzyme and thermophilic bacteria on the hydrolysis of mariculture organic waste (MOW).

    PubMed

    Guo, Liang; Sun, Mei; Zong, Yan; Zhao, Yangguo; Gao, Mengchun; She, Zonglian

    2016-01-01

    Mariculture organic waste (MOW) is rich in organic matter, which is a potential energy resource for anaerobic digestion. In order to enhance the anaerobic fermentation, the MOW was hydrolyzed by multi-enzyme and thermophilic bacteria. It was advantageous for soluble chemical oxygen demand (SCOD) release at MOW concentrations of 6 and 10 g/L with multi-enzyme and thermophilic bacteria pretreatments. For multi-enzyme, the hydrolysis was not obvious at substrate concentrations of 1 and 3 g/L, and the protein and carbohydrate increased with hydrolysis time at substrate concentrations of 6 and 10 g/L. For thermophilic bacteria, the carbohydrate was first released at 2-4 h and then consumed, and the protein increased with hydrolysis time. The optimal enzyme hydrolysis for MOW was determined by measuring the changes of SCOD, protein, carbohydrate, ammonia and total phosphorus, and comparing with acid and alkaline pretreatments.

  8. Microbial community structure in a thermophilic aerobic digester used as a sludge pretreatment process for the mesophilic anaerobic digestion and the enhancement of methane production.

    PubMed

    Jang, Hyun Min; Park, Sang Kyu; Ha, Jeong Hyub; Park, Jong Moon

    2013-10-01

    An effective two-stage sewage sludge digestion process, consisting of thermophilic aerobic digestion (TAD) followed by mesophilic anaerobic digestion (MAD), was developed for efficient sludge reduction and methane production. Using TAD as a biological pretreatment, the total volatile suspended solid reduction (VSSR) and methane production rate (MPR) in the MAD reactor were significantly improved. According to denaturing gradient gel electrophoresis (DGGE) analysis, the results indicated that the dominant bacteria species such as Ureibacillus thermophiles and Bacterium thermus in TAD were major routes for enhancing soluble organic matter. TAD pretreatment using a relatively short SRT of 1 day showed highly increased soluble organic products and positively affected an increment of bacteria populations which performed interrelated microbial metabolisms with methanogenic species in the MAD; consequently, a quantitative real-time PCR indicated greatly increased Methanosarcinales (acetate-utilizing methanogens) in the MAD, resulting in enhanced methane production. Copyright © 2013 Elsevier Ltd. All rights reserved.

  9. The chemical properties and microbial community characterization of the thermophilic microaerobic pretreatment process.

    PubMed

    Fu, Shan-Fei; He, Shuai; Shi, Xiao-Shuang; Katukuri, Naveen Reddy; Dai, Meng; Guo, Rong-Bo

    2015-12-01

    Thermophilic microaerobic pretreatment (TMP) was recently reported as an efficient pretreatment method of anaerobic digestion (AD). In this study, the chemical properties and microbial community were characterized to reveal how TMP working. Compared with thermophilic treatment under anaerobic condition (TMP0), cellulase activity obviously improved under microaerobic condition (TMP1), which was 10.9-49.0% higher than that of TMP0. Reducing sugar, SCOD and VFAs concentrations of TMP1 were 2.6-8.9%, 1.8-4.8% and 13.8-24% higher than those of TMP0, respectively. TMP gave obvious rise to phylum Firmicutes, which associated with extracellular enzymes production. The proportion of class Bacilli (belongs to phylum Firmicutes and mainly acts during hydrolysis) in TMP1 was 124.89% higher than that of TMP0, which reflected the greater hydrolytic ability under microaerobic condition. The improved abundance of phylum Firmicutes (especially class Bacilli, order Bacillales) under microaerobic condition could be the fundamental reason for the improved AD performance of thermophilic microaerobic pretreated corn straw. Copyright © 2015 Elsevier Ltd. All rights reserved.

  10. Enrichment of acetogenic bacteria in high rate anaerobic reactors under mesophilic and thermophilic conditions.

    PubMed

    Ryan, P; Forbes, C; McHugh, S; O'Reilly, C; Fleming, G T A; Colleran, E

    2010-07-01

    The objective of the current study was to expand the knowledge of the role of acetogenic Bacteria in high rate anaerobic digesters. To this end, acetogens were enriched by supplying a variety of acetogenic growth supportive substrates to two laboratory scale high rate upflow anaerobic sludge bed (UASB) reactors operated at 37 degrees C (R1) and 55 degrees C (R2). The reactors were initially fed a glucose/acetate influent. Having achieved high operational performance and granular sludge development and activity, both reactors were changed to homoacetogenic bacterial substrates on day 373 of the trial. The reactors were initially fed with sodium vanillate as a sole substrate. Although % COD removal indicated that the 55 degrees C reactor out performed the 37 degrees C reactor, effluent acetate levels from R2 were generally higher than from R1, reaching values as high as 5023 mg l(-1). Homoacetogenic activity in both reactors was confirmed on day 419 by specific acetogenic activity (SAA) measurement, with higher values obtained for R2 than R1. Sodium formate was introduced as sole substrate to both reactors on day 464. It was found that formate supported acetogenic activity at both temperatures. By the end of the trial, no specific methanogenic activity (SMA) was observed against acetate and propionate indicating that the methane produced was solely by hydrogenotrophic Archaea. Higher SMA and SAA values against H(2)/CO(2) suggested development of a formate utilising acetogenic population growing in syntrophy with hydrogenotrophic methanogens. Throughout the formate trial, the mesophilic reactor performed better overall than the thermophilic reactor. Copyright 2010 Elsevier Ltd. All rights reserved.

  11. Fertilizer potential of liquid and solid effluent from thermophilic anaerobic digestion of poultry waste.

    PubMed

    Liedl, B E; Bombardiere, J; Chaffield, J M

    2006-01-01

    Thermophilic anaerobic treatment of poultry litter produces an effluent stream of digested materials that can be separated into solid and liquid fractions for use as a crop fertilizer. The majority of the phosphorus is partitioned into the solid fraction while the majority of the nitrogen is present in the liquid fraction in the form of ammonium. These materials were tested over six years as an alternative fertilizer for the production of vegetable, fruit, and grassland crops. Application of the solids as a field crop fertilizer for vegetables and blueberries resulted in lower yields than the other fertilizer treatments, but an increase in soil phosphorus over a four-year period. Application of the digested liquids on grass and vegetable plots resulted in similar or superior yields to plots treated with commercially available nitrogen fertilizers. Hydroponic production of lettuce using liquid effluent was comparable to a commercial hydroponic fertilizer regime; however, the effluent treatment for hydroponic tomato production required supplementation and conversion of ammonium to nitrate. While not a total fertilizer solution, our research shows the effectiveness of digested effluent as part of a nutrient management program which could turn a livestock residuals problem into a crop nutrient resource.

  12. Defluviitalea raffinosedens sp. nov., a thermophilic, anaerobic, saccharolytic bacterium isolated from an anaerobic batch digester treating animal manure and rice straw.

    PubMed

    Ma, Shichun; Huang, Yan; Wang, Cong; Fan, Hui; Dai, Lirong; Zhou, Zheng; Liu, Xing; Deng, Yu

    2017-05-01

    A thermophilic, anaerobic, fermentative bacterium, strain A6T, was obtained from an anaerobic batch digester treating animal manure and rice straw. Cells were Gram-stain-positive, slightly curved rods with a size of 0.6-1×2.5-8.2 µm, non-motile and produced terminal spores. The temperature, pH and NaCl concentration ranges for growth were 40-60 °C, 6.5-8.0 and 0-15.0 g l-1, with optimum growth noted at 50-55 °C, pH 7.5 and in the absence of NaCl, respectively. Yeast extract was required for growth. d-Glucose, maltose, d-xylose, d-galactose, d-fructose, d-ribose, lactose, raffinose, sucrose, d-arabinose, cellobiose, d-mannose and yeast extract were used as carbon and energy sources. The fermentation products from glucose were ethanol, lactate, acetate, propionate, butyrate, valerate, iso-butyrate, iso-valerate, H2 and CO2. The G+C content of the genomic DNA was 36.6 mol%. The predominant fatty acids were C16 : 0, iso-C17 : 1, C14 : 0, C16 : 1ω7c, C16 : 0 N-alcohol and C13 : 0 3-OH. Respiratory quinones were not detected. The polar lipid profile comprised phosphoglycolipids, phospholipids, glycolipids, a diphosphatidylglycerol, a phosphatidylglycerol and an unidentified lipid. Phylogenetic analyses of the 16S rRNA gene sequence indicated that the strain was closely related to Defluviitalea saccharophila DSM 22681T with a similarity of 96.0 %. Based on the morphological, physiological and taxonomic characterization, strain A6T is considered to represent a novel species of the genus Defluviitalea, for which the name Defluviitalea raffinosedens sp. nov. is proposed. The type strain is A6T (=DSM 28090T=ACCC 19951T).

  13. Single-step ethanol production from lignocellulose using novel extremely thermophilic bacteria.

    PubMed

    Svetlitchnyi, Vitali A; Kensch, Oliver; Falkenhan, Doris A; Korseska, Svenja G; Lippert, Nadine; Prinz, Melanie; Sassi, Jamaleddine; Schickor, Anke; Curvers, Simon

    2013-02-28

    Consolidated bioprocessing (CBP) of lignocellulosic biomass to ethanol using thermophilic bacteria provides a promising solution for efficient lignocellulose conversion without the need for additional cellulolytic enzymes. Most studies on the thermophilic CBP concentrate on co-cultivation of the thermophilic cellulolytic bacterium Clostridium thermocellum with non-cellulolytic thermophilic anaerobes at temperatures of 55°C-60°C. We have specifically screened for cellulolytic bacteria growing at temperatures >70°C to enable direct conversion of lignocellulosic materials into ethanol. Seven new strains of extremely thermophilic anaerobic cellulolytic bacteria of the genus Caldicellulosiruptor and eight new strains of extremely thermophilic xylanolytic/saccharolytic bacteria of the genus Thermoanaerobacter isolated from environmental samples exhibited fast growth at 72°C, extensive lignocellulose degradation and high yield ethanol production on cellulose and pretreated lignocellulosic biomass. Monocultures of Caldicellulosiruptor strains degraded up to 89-97% of the cellulose and hemicellulose polymers in pretreated biomass and produced up to 72 mM ethanol on cellulose without addition of exogenous enzymes. In dual co-cultures of Caldicellulosiruptor strains with Thermoanaerobacter strains the ethanol concentrations rose 2- to 8.2-fold compared to cellulolytic monocultures. A co-culture of Caldicellulosiruptor DIB 087C and Thermoanaerobacter DIB 097X was particularly effective in the conversion of cellulose to ethanol, ethanol comprising 34.8 mol% of the total organic products. In contrast, a co-culture of Caldicellulosiruptor saccharolyticus DSM 8903 and Thermoanaerobacter mathranii subsp. mathranii DSM 11426 produced only low amounts of ethanol. The newly discovered Caldicellulosiruptor sp. strain DIB 004C was capable of producing unexpectedly large amounts of ethanol from lignocellulose in fermentors. The established co-cultures of new Caldicellulosiruptor

  14. Thermophilic anaerobic co-digestion of poultry litter and thin stillage.

    PubMed

    Sharma, Deepak; Espinosa-Solares, Teodoro; Huber, David H

    2013-05-01

    The purpose of this study was to test whether the performance of a thermophilic CSTR digester that has been stabilized on poultry litter will be enhanced or diminished by the addition of thin stillage as co-substrate. Replicate laboratory digesters, derived from a stable pilot-scale digester, were operated with increasing ratios (w/w) of thin stillage/poultry litter feedstock. After a period of adaptation to 20% and 40% thin stillage, digester performance showed increases in biogas, percent methane and COD removal, as well as a decrease in volatile acids. Peak performance occurred with 60% thin stillage. However, 80% thin stillage caused significant reduction of performance, including declines of methanogenic activity and COD removal. In conclusion, supplementing the thermophilic digestion of poultry litter with thin stillage improved the bioenergy (methane) output, but thin stillage became inhibitory at high concentrations. Copyright © 2013 Elsevier Ltd. All rights reserved.

  15. Retooling the ethanol industry: thermophilic anaerobic digestion of thin stillage for methane production and pollution prevention.

    PubMed

    Schaefer, Scott H; Sung, Shihwu

    2008-02-01

    Anaerobic digestion of corn ethanol thin stillage was tested at thermophilic temperature (55 degrees C) with two completely stirred tank reactors. The thin stillage wastestream was organically concentrated with 100 g/L total chemical oxygen demand and 60 g/L volatiles solids and a low pH of approximately 4.0. Steady-state was achieved at 30-, 20-, and 15-day hydraulic retention times (HRTs) and digester failure at a 12-day HRT. Significant reduction of volatile solids was achieved, with a maximum reduction (89.8%) at the 20-day HRT. Methane yield ranged from 0.6 to 0.7 L methane/g volatile solids removed during steady-state operation. Effluent volatile fatty acids below 200 mg/L as acetic acid were achieved at 20- and 30-day HRTs. Ultrasonic pretreatment was used for one digester, although no significant improvement was observed. Ethanol plant natural gas consumption could be reduced 43 to 59% with the methane produced, while saving an estimated $7 to $17 million ($10 million likely) for a facility producing 360 million L ethanol/y.

  16. Characterization of cellulolytic enzymes and bioH2 production from anaerobic thermophilic Clostridium sp. TCW1.

    PubMed

    Lo, Yung-Chung; Huang, Chi-Yu; Cheng, Chieh-Lun; Lin, Chiu-Yue; Chang, Jo-Shu

    2011-09-01

    A thermophilic anaerobic bacterium Clostridium sp. TCW1 was isolated from dairy cow dung and was used to produce hydrogen from cellulosic feedstock. Extracellular cellulolytic enzymes produced from TCW1 strain were identified as endoglucanases (45, 53 and 70 kDa), exoglucanase (70 kDa), xylanases (53 and 60 kDa), and β-glucosidase (45 kDa). The endoglucanase and xylanase were more abundant. The optimal conditions for H2 production and enzyme production of the TCW1 strain were the same (60 °C, initial pH 7, agitation rate of 200 rpm). Ten cellulosic feedstock, including pure or natural cellulosic materials, were used as feedstock for hydrogen production by Clostridium strain TCW1 under optimal culture conditions. Using filter paper at 5.0 g/L resulted in the most effective hydrogen production performance, achieving a H2 production rate and yield of 57.7 ml/h/L and 2.03 mol H2/mol hexose, respectively. Production of cellulolytic enzyme activities was positively correlated with the efficiency of dark-H2 fermentation. Copyright © 2011 Elsevier Ltd. All rights reserved.

  17. Evaluation of Integrated Anaerobic Digestion and Hydrothermal Carbonization for Bioenergy Production

    PubMed Central

    Reza, M. Toufiq; Werner, Maja; Pohl, Marcel; Mumme, Jan

    2014-01-01

    Lignocellulosic biomass is one of the most abundant yet underutilized renewable energy resources. Both anaerobic digestion (AD) and hydrothermal carbonization (HTC) are promising technologies for bioenergy production from biomass in terms of biogas and HTC biochar, respectively. In this study, the combination of AD and HTC is proposed to increase overall bioenergy production. Wheat straw was anaerobically digested in a novel upflow anaerobic solid state reactor (UASS) in both mesophilic (37 °C) and thermophilic (55 °C) conditions. Wet digested from thermophilic AD was hydrothermally carbonized at 230 °C for 6 hr for HTC biochar production. At thermophilic temperature, the UASS system yields an average of 165 LCH4/kgVS (VS: volatile solids) and 121 L CH4/kgVS at mesophilic AD over the continuous operation of 200 days. Meanwhile, 43.4 g of HTC biochar with 29.6 MJ/kgdry_biochar was obtained from HTC of 1 kg digestate (dry basis) from mesophilic AD. The combination of AD and HTC, in this particular set of experiment yield 13.2 MJ of energy per 1 kg of dry wheat straw, which is at least 20% higher than HTC alone and 60.2% higher than AD only. PMID:24962786

  18. Comparison of thermophilic bacteria and alkyl polyglucose pretreatment on two-stage anaerobic digestion with waste sludge: Biogas production potential and substrate metabolism process.

    PubMed

    Guo, Liang; Zhang, Zengshuai; Gao, Mengchun; She, Zonglian; Zhao, Yangguo; Guo, Yiding; Sun, Jian

    2018-02-01

    To gain a better understanding of the influence on two-stage anaerobic digestion of waste sludge with thermophilic bacteria (TB) and alkyl polyglucose (APG) pretreatment, changing of soluble chemical oxygen demand (SCOD), carbohydrate and protein in extracellular polymeric substances (EPS) and dissolved organic matters (DOM) were analyzed. The excitation-emission matrix (EEM) with fluorescence regional integration (FRI) was also used to investigate compositional and structural characteristics of DOM. The highest hydrogen and methane yield of TB pretreated sludge was 12.2 ml/g VS (volatile suspended solid) and 124.7 ml/g VS, and that of APG pretreated sludge was 28.3 ml/g VS and 19.9 ml/g VS. The VS removal of TB pretreated sludge (36.7%) was higher than APG pretreated sludge (27.1%) in the two-stage anaerobic digestion. The APG pretreatment could inhibit the activity of methanogens and the substrate (such as volatile fatty acids (VFAs), protein and soluble microbial materials) was accumulated compared with TB pretreatment. Copyright © 2017 Elsevier Ltd. All rights reserved.

  19. Characterization of Melioribacter roseus gen. nov., sp. nov., a novel facultatively anaerobic thermophilic cellulolytic bacterium from the class Ignavibacteria, and a proposal of a novel bacterial phylum Ignavibacteriae.

    PubMed

    Podosokorskaya, Olga A; Kadnikov, Vitaly V; Gavrilov, Sergey N; Mardanov, Andrey V; Merkel, Alexander Y; Karnachuk, Olga V; Ravin, Nikolay V; Bonch-Osmolovskaya, Elizaveta A; Kublanov, Ilya V

    2013-06-01

    A novel moderately thermophilic, facultatively anaerobic chemoorganotrophic bacterium strain P3M-2(T) was isolated from a microbial mat developing on the wooden surface of a chute under the flow of hot water (46°C) coming out of a 2775-m-deep oil exploration well (Tomsk region, Russia). Strain P3M-2(T) is a moderate thermophile and facultative anaerobe growing on mono-, di- or polysaccharides by aerobic respiration, fermentation or by reducing diverse electron acceptors [nitrite, Fe(III), As(V)]. Its closest cultivated relative (90.8% rRNA gene sequence identity) is Ignavibacterium album, the only chemoorganotrophic member of the phylum Chlorobi. New genus and species Melioribacter roseus are proposed for isolate P3M-2(T) . Together with I. album, the new organism represents the class Ignavibacteria assigned to the phylum Chlorobi. The revealed group includes a variety of uncultured environmental clones, the 16S rRNA gene sequences of some of which have been previously attributed to the candidate division ZB1. Phylogenetic analysis of M. roseus and I. album based on their 23S rRNA and RecA sequences confirmed that these two organisms could represent an even deeper, phylum-level lineage. Hence, we propose a new phylum Ignavibacteriae within the Bacteroidetes-Chlorobi group with a sole class Ignavibacteria, two families Ignavibacteriaceae and Melioribacteraceae and two species I. album and M. roseus. This proposal correlates with chemotaxonomic data and phenotypic differences of both organisms from other cultured representatives of Chlorobi. The most essential differences, supported by the analyses of complete genomes of both organisms, are motility, facultatively anaerobic and obligately organotrophic mode of life, the absence of chlorosomes and the apparent inability to grow phototrophically. © 2012 Society for Applied Microbiology and Blackwell Publishing Ltd.

  20. Microbial examination of anaerobic sludge adaptation to animal slurry.

    PubMed

    Moset, V; Cerisuelo, A; Ferrer, P; Jimenez, A; Bertolini, E; Cambra-López, M

    2014-01-01

    The objective of this study was to evaluate changes in the microbial population of anaerobic sludge digesters during the adaptation to pig slurry (PS) using quantitative real-time polymerase chain reaction (qPCR) and qualitative scanning electron microscopy (SEM). Additionally, the relationship between microbial parameters and sludge physicochemical composition and methane yield was examined. Results showed that the addition of PS to an unadapted thermophilic anaerobic digester caused an increase in volatile fatty acids (VFA) concentration, a decrease in removal efficiency and CH4 yield. Additionally, increases in total bacteria and total archaea were observed using qPCR. Scanning electron micrographs provided a general overview of the sludge's cell morphology, morphological diversity and degree of organic matter degradation. A change in microbial morphotypes from homogeneous cell morphologies to a higher morphological diversity, similar to that observed in PS, was observed with the addition of PS by SEM. Therefore, the combination of qPCR and SEM allowed expanding the knowledge about the microbial adaptation to animal slurry in thermophilic anaerobic digesters.

  1. In Situ Expression of Acidic and Thermophilic Carbohydrate Active Enzymes by Filamentous Fungi (JGI Seventh Annual User Meeting 2012: Genomics of Energy and Environment)

    ScienceCinema

    Mosier, Annika

    2018-01-22

    Annika Mosier, graduate student from Stanford University presents a talk titled "In Situ Expression of Acidic and Thermophilic Carbohydrate Active Enzymes by Filamentous Fungi" at the JGI User 7th Annual Genomics of Energy & Environment Meeting on March 22, 2012 in Walnut Creek, CA.

  2. Evaluation of the rotary drum reactor process as pretreatment technology of municipal solid waste for thermophilic anaerobic digestion and biogas production.

    PubMed

    Gikas, Petros; Zhu, Baoning; Batistatos, Nicolas Ion; Zhang, Ruihong

    2018-06-15

    Municipal solid waste (MSW) contains a large fraction of biodegradable organic materials. When disposed in landfills, these materials can cause adverse environmental impact due to gaseous emissions and leachate generation. This study was performed with an aim of effectively separating the biodegradable materials from a Mechanical Biological Treatment (MBT) facility and treating them in well-controlled anaerobic digesters for biogas production. The rotary drum reactor (RDR) process (a sub-process of the MBT facilities studied in the present work) was evaluated as an MSW pretreatment technology for separating and preparing the biodegradable materials in MSW to be used as feedstock for anaerobic digestion. The RDR processes used in six commercial MSW treatment plants located in the USA were surveyed and sampled. The samples of the biodegradable materials produced by the RDR process were analyzed for chemical and physical characteristics as well as anaerobically digested in the laboratory using batch reactors under thermophilic conditions. The moisture content, TS, VS and C/N of the samples varied between 64.7 and 44.4%, 55.6 to 35.3%, 27.0 to 41.3% and 24.5 to 42.7, respectively. The biogas yield was measured to be between 533.0 and 675.6 mL g -1 VS after 20 days of digestion. Approximately 90% of the biogas was produced during the first 13 days. The average methane content of the biogas was between 58.0 and 59.9%. The results indicated that the biodegradable materials separated from MSW using the RDR processes could be used as an excellent feedstock for anaerobic digestion. The digester residues may be further processed for compost production or further energy recovery by using thermal conversion processes such as combustion or gasification. Copyright © 2017. Published by Elsevier Ltd.

  3. Bioflocculation of mesophilic and thermophilic activated sludge.

    PubMed

    Vogelaar, J C T; De Keizer, A; Spijker, S; Lettinga, G

    2005-01-01

    Thermophilic activated sludge treatment is often hampered by a turbid effluent. Reasons for this phenomenon are so far unknown. Here, the hypothesis of the temperature dependency of the hydrophobic interaction as a possible cause for diminished thermophilic activated sludge bioflocculation was tested. Adsorption of wastewater colloidal particles was monitored on different flat surfaces as a function of temperature. Adsorption on a hydrophobic surface varied with temperature between 20 and 60 degrees C and no upward or downward trend could be observed. This makes the hydrophobic interaction hypothesis unlikely in explaining the differences in mesophilic and thermophilic activated sludge bioflocculation. Both mesophilic and thermophilic biomass did not flocculate with wastewater colloidal particles under anaerobic conditions. Only in the presence of oxygen, with biologically active bacteria, the differences in bioflocculation behavior became evident. Bioflocculation was shown only to occur with the combination of wastewater and viable mesophilic biomass at 30 degrees C, in the presence of oxygen. Bioflocculation did not occur in case the biomass was inactivated or when oxygen was absent. Thermophilic activated sludge hardly showed any bioflocculation, also under mesophilic conditions. Despite the differences in bioflocculation behavior, sludge hydrophobicity and sludge zetapotentials were almost similar. Theoretical calculations using the DLVO (Derjaguin, Landau, Verweij and Overbeek) theory showed that flocculation is unlikely in all cases due to long-range electrostatic forces. These calculations, combined with the fact that bioflocculation actually did occur at 30 degrees C and the unlikelyness of the hydrophobic interaction, point in the direction of bacterial exo-polymers governing bridging flocculation. Polymer interactions are not included in the DLVO theory and may vary as a function of temperature.

  4. Molecular analysis of meso- and thermophilic microbiota associated with anaerobic biowaste degradation

    PubMed Central

    2012-01-01

    Background Microbial anaerobic digestion (AD) is used as a waste treatment process to degrade complex organic compounds into methane. The archaeal and bacterial taxa involved in AD are well known, whereas composition of the fungal community in the process has been less studied. The present study aimed to reveal the composition of archaeal, bacterial and fungal communities in response to increasing organic loading in mesophilic and thermophilic AD processes by applying 454 amplicon sequencing technology. Furthermore, a DNA microarray method was evaluated in order to develop a tool for monitoring the microbiological status of AD. Results The 454 sequencing showed that the diversity and number of bacterial taxa decreased with increasing organic load, while archaeal i.e. methanogenic taxa remained more constant. The number and diversity of fungal taxa increased during the process and varied less in composition with process temperature than bacterial and archaeal taxa, even though the fungal diversity increased with temperature as well. Evaluation of the microarray using AD sample DNA showed correlation of signal intensities with sequence read numbers of corresponding target groups. The sensitivity of the test was found to be about 1%. Conclusions The fungal community survives in anoxic conditions and grows with increasing organic loading, suggesting that Fungi may contribute to the digestion by metabolising organic nutrients for bacterial and methanogenic groups. The microarray proof of principle tests suggest that the method has the potential for semiquantitative detection of target microbial groups given that comprehensive sequence data is available for probe design. PMID:22727142

  5. Quantification of viable but nonculturable Salmonella spp. and Shigella spp. during sludge anaerobic digestion and their reactivation during cake storage.

    PubMed

    Fu, B; Jiang, Q; Liu, H-B; Liu, H

    2015-10-01

    The presence of viable but nonculturable (VBNC) bacterial pathogens which often fail to be detected by cultivation and can regain the cultivability if the living conditions improve were reported. The objective of this study was to determine the occurrence of VBNC Salmonella spp. and Shigella spp. in the biosolids during anaerobic digestion and its reactivation during the cake storage. The occurrence of VBNC Salmonella spp. and Shigella spp. during mesophilic, temperature-phased, thermophilic anaerobic digestion of sewage sludge and the subsequent storage were studied by RT-qPCR and most probable number (MPN) method. The VBNC incidence of Salmonella spp. and Shigella spp. during thermophilic digestion was four orders of magnitude higher than those of mesophilic digestion. Accordingly, higher resuscitation ratio of VBNC pathogens was also achieved in thermophilic digested sludge. As a result, the culturable Salmonella typhimurium contents in thermophilic digested sludge after cake storage were two orders of magnitude higher than mesophilic digestion. Both quantitative PCR and reverse transcription quantitative PCR assay results showed the two bacterial counting numbers remained stable throughout the cake storage. The results indicate that the increase in the culturable Salmonella spp. and Shigella spp. after centrifugal dewatering was attributed to the resuscitation from the VBNC state to the culturable state. Thermophilic anaerobic digestion mainly induced Salmonella spp. and Shigella spp. into VBNC state rather than killed them, suggesting that the biological safety of sewage sludge by temperature-phased anaerobic digestion should be carefully assessed. © 2015 The Society for Applied Microbiology.

  6. Inter-stage thermophilic aerobic digestion may increase organic matter removal from wastewater sludge without decreasing biogas production.

    PubMed

    Hafner, Sasha D; Madsen, Johan T; Pedersen, Johanna M; Rennuit, Charlotte

    2018-02-01

    Combining aerobic and anaerobic digestion in a two-stage system can improve the degradation of wastewater sludge over the use of either technology alone. But use of aerobic digestion as a pre-treatment before anaerobic digestion generally reduces methane production due to loss of substrate through oxidation. An inter-stage configuration may avoid this reduction in methane production. Here, we evaluated the use of thermophilic aerobic digestion (TAD) as an inter-stage treatment for wastewater sludge using laboratory-scale semi-continuous reactors. A single anaerobic digester was compared to an inter-stage system, where a thermophilic aerobic digester (55 °C) was used between two mesophilic anaerobic digesters (37 °C). Both systems had retention times of approximately 30 days, and the comparison was based on measurements made over 97 days. Results showed that the inter-stage system provided better sludge destruction (52% volatile solids (VS) removal vs. 40% for the single-stage system, 44% chemical oxygen demand (COD) removal vs. 34%) without a decrease in total biogas production (methane yield per g VS added was 0.22-0.24 L g -1 for both systems).

  7. Comparison of microbial communities during the anaerobic digestion of Gracilaria under mesophilic and thermophilic conditions.

    PubMed

    Azizi, Aqil; Kim, Wonduck; Lee, Jung Hyun

    2016-10-01

    Mesophilic and thermophilic anaerobic digesters (MD and TD, respectively) utilizing Gracilaria and marine sediment as the substrate and inoculum, respectively, were compared by analyzing their performances and microbial community changes. During three successive transfers, the average cumulative methane yields in the MD and TD were 222.6 ± 17.3 mL CH4/g volatile solids (VS) and 246.1 ± 11 mL CH4/g VS, respectively. The higher hydrolysis rate and acidogenesis in the TD resulted in a several fold greater accumulation of volatile fatty acids (acetate, propionate, and butyrate) followed by a larger pH drop with a prolonged recovery than in the MD. However, the operational stability between both digesters remained comparable. Pyrosequencing analyses revealed that the MD had more complex microbial diversity indices and microbial community changes than the TD. Interestingly, Methanomassiliicoccales, the seventh methanogen order was the predominant archaeal order in the MD along with bacterial orders of Clostridiales, Bacteriodales, and Synergistales. Meanwhile, Coprothermobacter and Methanobacteriales dominated the bacterial and archaeal community in the TD, respectively. Although the methane yield is comparable, both MD and TD show a different profile of pH, VFA and the microbial communities.

  8. Isolation, identification and utilization of thermophilic strains in aerobic digestion of sewage sludge.

    PubMed

    Liu, Shugen; Zhu, Nanwen; Li, Loretta Y; Yuan, Haiping

    2011-11-15

    Two representative thermophilic bacterial strains (T1 and T2) were isolated from a one-stage autothermal thermophilic aerobic digestion pilot-scale reactor. 16S rRNA gene analysis indicated that they were Hydrogenophilaceae and Xanthomonodaceae. These isolated strains were inoculated separately and/or jointly in sewage sludge, to investigate their effects on sludge stabilization under thermophilic aerobic digestion condition. Four digestion conditions were tested for 480 h. Digestion without inoculation and inoculation with strain T2, as well as joint- inoculation with strains T1 and T2, achieved 32.6%, 43.0%, and 38.2% volatile solids (VS) removal, respectively. Removal in a digester inoculated with stain T1 only reached 27.2%. For the first 144 h, the three inoculated digesters all experienced higher VS removal than the digester without inoculations. Both specific thermophilic strains and micro-environment significantly affected the VS removal. DGGE profiles revealed that the isolated strains T1 and T2 can successfully establish in the thermophilic digesters. Other viable bacteria (including anaerobic or facultative microbes) also appeared in the digestion system, enhancing the microbial activity. Copyright © 2011. Published by Elsevier Ltd.

  9. Temperature-phased anaerobic digestion of food waste: A comparison with single-stage digestions based on performance and energy balance.

    PubMed

    Xiao, Benyi; Qin, Yu; Zhang, Wenzhe; Wu, Jing; Qiang, Hong; Liu, Junxin; Li, Yu-You

    2018-02-01

    The temperature-phased anaerobic digestion (TPAD) of food waste was studied for the purpose of comparing with single-stage mesophilic and thermophilic anaerobic digestion. The biogas and methane yields in the TPAD during the steady period were 0.759 ± 0.115 L/g added VS and 0.454 ± 0.201 L/g added VS, which were lower than those in the two single-stage anaerobic digestion. The improper sludge retention time may be the reason for the lower biogas and methane production in TPAD. The removal of volatile solids in the TPAD was 78.55 ± 4.59% and the lowest among the three anaerobic digestion processes. The reaction ratios of the four anaerobic digestion steps in the TPAD were all lower than those in the two single-stage anaerobic digestion. The energy conversion efficiency of the degraded substrate in the TPAD was similar with those in single-stage mesophilic and thermophilic anaerobic digestion systems. Copyright © 2017 Elsevier Ltd. All rights reserved.

  10. Influence of high gas production during thermophilic anaerobic digestion in pilot-scale and lab-scale reactors on survival of the thermotolerant pathogens Clostridium perfringens and Campylobacter jejuni in piggery wastewater.

    PubMed

    Skillman, L C; Bajsa, O; Ho, L; Santhanam, B; Kumar, M; Ho, G

    2009-07-01

    Safe reuse of animal wastes to capture energy and nutrients, through anaerobic digestion processes, is becoming an increasingly desirable solution to environmental pollution. Pathogen decay is the most important safety consideration and is in general, improved at elevated temperatures and longer hydraulic residence times. During routine sampling to assess pathogen decay in thermophilic digestion, an inversely proportional relationship between levels of Clostridium perfringens and gas production was observed. Further samples were collected from pilot-scale, bench-scale thermophilic reactors and batch scale vials to assess whether gas production (predominantly methane) could be a useful indicator of decay of the thermotolerant pathogens C. perfringens and Campylobacter jejuni. Pathogen levels did appear to be lower where gas production and levels of methanogens were higher. This was evident at each operating temperature (50, 57, 65 degrees C) in the pilot-scale thermophilic digesters, although higher temperatures also reduced the numbers of pathogens detected. When methane production was higher, either when feed rate was increased, or pH was lowered from 8.2 (piggery wastewater) to 6.5, lower numbers of pathogens were detected. Although a number of related factors are known to influence the amount and rate of methane production, it may be a useful indicator of the removal of the pathogens C. perfringens and C. jejuni.

  11. Microbial monitoring by molecular tools of an upflow anaerobic filter treating abattoir wastewaters.

    PubMed

    Gannoun, Hana; Khelifi, Eltaief; Omri, Ilhem; Jabari, Linda; Fardeau, Marie-Laure; Bouallagui, Hassib; Godon, Jean-Jacques; Hamdi, Moktar

    2013-08-01

    The performance of anaerobic digestion of abattoir wastewaters (AW) in an upflow anaerobic filter (UAF) was investigated under mesophilic (37°C) and thermophilic (55°C) conditions. The effects of increasing temperature on the performance of the UAF and on the dynamics of the microbial community of the anaerobic sludge were studied. The results showed that chemical oxygen demand (COD) removal efficiency of 90% was achieved for organic loading rates (OLRs) up to 4.5g CODL(-1)d(-1) in mesophilic conditions, while in thermophilic conditions, the highest OLRs of 9 g CODL(-1)d(-1) led to the efficiency of 72%. The use of molecular and microbiological methods to recover microbial populations involved in this process showed that fermentative bacteria were the prominent members of the sludge microbial community. Three novel strains were identified as Macellibacteroides fermentans, Desulfotomaculum peckii and Defluviitalea saccharophila. Copyright © 2013 Elsevier Ltd. All rights reserved.

  12. Anaerobic Codigestion of Sludge: Addition of Butcher's Fat Waste as a Cosubstrate for Increasing Biogas Production.

    PubMed

    Martínez, E J; Gil, M V; Fernandez, C; Rosas, J G; Gómez, X

    2016-01-01

    Fat waste discarded from butcheries was used as a cosubstrate in the anaerobic codigestion of sewage sludge (SS). The process was evaluated under mesophilic and thermophilic conditions. The codigestion was successfully attained despite some inhibitory stages initially present that had their origin in the accumulation of volatile fatty acids (VFA) and adsorption of long-chain fatty acids (LCFA). The addition of a fat waste improved digestion stability and increased biogas yields thanks to the higher organic loading rate (OLR) applied to the reactors. However, thermophilic digestion was characterized by an effluent of poor quality and high VFA content. Results from spectroscopic analysis suggested the adsorption of lipid components onto the anaerobic biomass, thus disturbing the complete degradation of substrate during the treatment. The formation of fatty aggregates in the thermophilic reactor prevented process failure by avoiding the exposure of biomass to the toxic effect of high LCFA concentrations.

  13. Volatile fatty acids productions by mesophilic and thermophilic sludge fermentation: Biological responses to fermentation temperature.

    PubMed

    Hao, Jiuxiao; Wang, Hui

    2015-01-01

    The volatile fatty acids (VFAs) productions, as well as hydrolases activities, microbial communities, and homoacetogens, of mesophilic and thermophilic sludge anaerobic fermentation were investigated to reveal the microbial responses to different fermentation temperatures. Thermophilic fermentation led to 10-fold more accumulation of VFAs compared to mesophilic fermentation. α-glucosidase and protease had much higher activities in thermophilic reactor, especially protease. Illumina sequencing manifested that raising fermentation temperature increased the abundances of Clostridiaceae, Microthrixaceae and Thermotogaceae, which could facilitate either hydrolysis or acidification. Real-time PCR analysis demonstrated that under thermophilic condition the relative abundance of homoacetogens increased in batch tests and reached higher level at stable fermentation, whereas under mesophilic condition it only increased slightly in batch tests. Therefore, higher fermentation temperature increased the activities of key hydrolases, raised the proportions of bacteria involved in hydrolysis and acidification, and promoted the relative abundance of homoacetogens, which all resulted in higher VFAs production. Copyright © 2014 Elsevier Ltd. All rights reserved.

  14. Draft Genome Sequence of an Anaerobic and Extremophilic Bacterium, Caldanaerobacter yonseiensis, Isolated from a Geothermal Hot Stream

    PubMed Central

    Lee, Sang-Jae; Lee, Yong-Jik; Park, Gun-Seok; Kim, Byoung-Chan; Lee, Sang Jun; Shin, Jae-Ho

    2013-01-01

    Caldanaerobacter yonseiensis is a strictly anaerobic, thermophilic, spore-forming bacterium, which was isolated from a geothermal hot stream in Indonesia. This bacterium utilizes xylose and produces a variety of proteases. Here, we report the draft genome sequence of C. yonseiensis, which reveals insights into the pentose phosphate pathway and protein degradation metabolism in thermophilic microorganisms. PMID:24201201

  15. Potential Application of Anaerobic Extremophiles for Hydrogen Production

    NASA Technical Reports Server (NTRS)

    Pikuta, Elena V.; Hoover, Richard B.

    2004-01-01

    During substrate fermentation many anaerobes produce the hydrogen as a waste product, which often regulates the growth of the cultures as an inhibitor. In nature the hydrogen is usually removed from the ecosystem due to its physical properties or by consumption of hydrogen by secondary anaerobes, which sometimes behave as competitors for electron donors as is seen in the classical example in anaerobic microbial communities via the interaction between methanogens and sulfate- or sulfur- reducers. It was demonstrated previously on mixed cultures of anaerobes at neutral pH that bacterial hydrogen production could provide an alternative energy source. But at neutral pH the original cultures can easily be contaminated by methanogens, a most unpleasant side effect of these conditions is the development of pathogenic bacteria. In both cases the rate of hydrogen production was dramatically decreased since some part of the hydrogen was transformed to methane, and the cultivation of human pathogens on a global scale is very dangerous. In our laboratory, experiments with obligately alkaliphilic bacteria that excrete hydrogen as the end metabolic product were performed at different temperature regimes. Mesophilic and moderately thermophilic bacterial cultures have been studied and compared for the most effective hydrogen production. For high-mineralized media with pH 9.5-10.0 not many methanogens are known to exist. Furthermore, the development of pathogenic contaminant microorganisms is virtually impossible: carbonate-saturated solutions are used as antiseptics in medicine. Therefore the cultivation of alkaliphilic hydrogen producing bacteria could be considered as most safe process for global Scale industry in future. Here we present experimental data on the rates of hydrogen productivity for mesophilic, alkaliphilic, obligately anaerobic bacterium Spirocheta americana ASpG1 and moderately thermophilic, alkaliphilic, facultative anaerobe Anoxybacillus pushchinoensis K1 and

  16. Temperature phased anaerobic digestion increases apparent hydrolysis rate for waste activated sludge.

    PubMed

    Ge, Huoqing; Jensen, Paul D; Batstone, Damien J

    2011-02-01

    It is well established that waste activated sludge with an extended sludge age is inherently slow to degrade with a low extent of degradation. Pre-treatment methods can be used prior to anaerobic digestion to improve the efficiency of activated sludge digestion. Among these pre-treatment methods, temperature phased anaerobic digestion (TPAD) is one promising method with a relatively low energy input and capital cost. In this study, an experimental thermophilic (50-70 °C)-mesophilic system was compared against a control mesophilic-mesophilic system. The thermophilic-mesophilic system achieved 41% and 48% volatile solids (VS) destruction during pre-treatment of 60 °C and 65 °C (or 70 °C) respectively, compared to 37% in the mesophilic-mesophilic TPAD system. Solubilisation in the first stage was enhanced during thermophilic pre-treatment (15% at 50 °C and 27% at 60 °C, 65 °C and 70 °C) over mesophilic pre-treatment (7%) according to a COD balance. This was supported by ammonia-nitrogen measurements. Model based analysis indicated that the mechanism for increased performance was due to an increase in hydrolysis coefficient under thermophilic pre-treatment of 60 °C (0.5 ± 0.1 d(-1)), 65 °C (0.7 ± 0.2 d(-1)) and 70 °C (0.8 ± 0.2 d(-1)) over mesophilic pre-treatment (0.2 ± 0.1 d(-1)), and thermophilic pre-treatment at 50 °C (0.12 ± 0.06 d(-1)). © 2010 Elsevier Ltd. All rights reserved.

  17. Mass and Energy Balances of Dry Thermophilic Anaerobic Digestion Treating Swine Manure Mixed with Rice Straw.

    PubMed

    Zhou, Sheng; Zhang, Jining; Zou, Guoyan; Riya, Shohei; Hosomi, Masaaki

    2015-01-01

    To evaluate the feasibility of swine manure treatment by a proposed Dry Thermophilic Anaerobic Digestion (DT-AD) system, we evaluated the methane yield of swine manure treated using a DT-AD method with rice straw under different C/N ratios and solid retention time (SRT) and calculated the mass and energy balances when the DT-AD system is used for swine manure treatment from a model farm with 1000 pigs and the digested residue is used for forage rice production. A traditional swine manure treatment Oxidation Ditch system was used as the study control. The results suggest that methane yield using the proposed DT-AD system increased with a higher C/N ratio and shorter SRT. Correspondently, for the DT-AD system running with SRT of 80 days, the net energy yields for all treatments were negative, due to low biogas production and high heat loss of digestion tank. However, the biogas yield increased when the SRT was shortened to 40 days, and the generated energy was greater than consumed energy when C/N ratio was 20 : 1 and 30 : 1. The results suggest that with the correct optimization of C/N ratio and SRT, the proposed DT-AD system, followed by using digestate for forage rice production, can attain energy self-sufficiency.

  18. Mass and Energy Balances of Dry Thermophilic Anaerobic Digestion Treating Swine Manure Mixed with Rice Straw

    PubMed Central

    Zhou, Sheng; Zhang, Jining; Zou, Guoyan; Riya, Shohei; Hosomi, Masaaki

    2015-01-01

    To evaluate the feasibility of swine manure treatment by a proposed Dry Thermophilic Anaerobic Digestion (DT-AD) system, we evaluated the methane yield of swine manure treated using a DT-AD method with rice straw under different C/N ratios and solid retention time (SRT) and calculated the mass and energy balances when the DT-AD system is used for swine manure treatment from a model farm with 1000 pigs and the digested residue is used for forage rice production. A traditional swine manure treatment Oxidation Ditch system was used as the study control. The results suggest that methane yield using the proposed DT-AD system increased with a higher C/N ratio and shorter SRT. Correspondently, for the DT-AD system running with SRT of 80 days, the net energy yields for all treatments were negative, due to low biogas production and high heat loss of digestion tank. However, the biogas yield increased when the SRT was shortened to 40 days, and the generated energy was greater than consumed energy when C/N ratio was 20 : 1 and 30 : 1. The results suggest that with the correct optimization of C/N ratio and SRT, the proposed DT-AD system, followed by using digestate for forage rice production, can attain energy self-sufficiency. PMID:26609436

  19. Effect of long residence time and high temperature over anaerobic biodegradation of Scenedesmus microalgae grown in wastewater.

    PubMed

    Greses, S; Zamorano-López, N; Borrás, L; Ferrer, J; Seco, A; Aguado, D

    2018-07-15

    Anaerobic digestion of indigenous Scenedesmus spp. microalgae was studied in continuous lab-scale anaerobic reactors at different temperatures (35 °C and 55 °C), and sludge retention time - SRT (50 and 70 days). Mesophilic digestion was performed in a continuous stirred-tank reactor (CSTR) and in an anaerobic membrane bioreactor (AnMBR). Mesophilic CSTR operated at 50 days SRT only achieved 11.9% of anaerobic biodegradability whereas in the AnMBR at 70 days SRT and 50 days HRT reached 39.5%, which is even higher than the biodegradability achieved in the thermophilic CSTR at 50 days SRT (30.4%). Microbial analysis revealed a high abundance of cellulose-degraders in both reactors, AnMBR (mainly composed of 9.4% Bacteroidetes, 10.1% Chloroflexi, 8.0% Firmicutes and 13.2% Thermotogae) and thermophilic CSTR (dominated by 23.8% Chloroflexi and 12.9% Firmicutes). However, higher microbial diversity was found in the AnMBR compared to the thermophilic CSTR which is related to the SRT. since high SRT promoted low growth-rate microorganisms, increasing the hydrolytic potential of the system. These results present the membrane technology as a promising approach to revalue microalgal biomass, suggesting that microalgae biodegradability and consequently the methane production could be improved operating at higher SRT. Copyright © 2018 Elsevier Ltd. All rights reserved.

  20. Anaerobic Codigestion of Sludge: Addition of Butcher’s Fat Waste as a Cosubstrate for Increasing Biogas Production

    PubMed Central

    Martínez, E. J.; Gil, M. V.; Fernandez, C.; Rosas, J. G.

    2016-01-01

    Fat waste discarded from butcheries was used as a cosubstrate in the anaerobic codigestion of sewage sludge (SS). The process was evaluated under mesophilic and thermophilic conditions. The codigestion was successfully attained despite some inhibitory stages initially present that had their origin in the accumulation of volatile fatty acids (VFA) and adsorption of long-chain fatty acids (LCFA). The addition of a fat waste improved digestion stability and increased biogas yields thanks to the higher organic loading rate (OLR) applied to the reactors. However, thermophilic digestion was characterized by an effluent of poor quality and high VFA content. Results from spectroscopic analysis suggested the adsorption of lipid components onto the anaerobic biomass, thus disturbing the complete degradation of substrate during the treatment. The formation of fatty aggregates in the thermophilic reactor prevented process failure by avoiding the exposure of biomass to the toxic effect of high LCFA concentrations. PMID:27071074

  1. Comparison of bacterial community structure and dynamics during the thermophilic composting of different types of solid wastes: anaerobic digestion residue, pig manure and chicken manure.

    PubMed

    Song, Caihong; Li, Mingxiao; Jia, Xuan; Wei, Zimin; Zhao, Yue; Xi, Beidou; Zhu, Chaowei; Liu, Dongming

    2014-09-01

    This study investigated the impact of composting substrate types on the bacterial community structure and dynamics during composting processes. To this end, pig manure (PM), chicken manure (CM), a mixture of PM and CM (PM + CM), and a mixture of PM, CM and anaerobic digestion residue (ADR) (PM + CM + ADR) were selected for thermophilic composting. The bacterial community structure and dynamics during the composting process were detected and analysed by polymerase chain reaction-denaturing gradient gel electrophoresis (DGGE) coupled with a statistic analysis. The physical-chemical analyses indicated that compared to single-material composting (PM, CM), co-composting (PM + CM, PM + CM + ADR) could promote the degradation of organic matter and strengthen the ability of conserving nitrogen. A DGGE profile and statistical analysis demonstrated that co-composting, especially PM + CM + ADR, could improve the bacterial community structure and functional diversity, even in the thermophilic stage. Therefore, co-composting could weaken the screening effect of high temperature on bacterial communities. Dominant sequencing analyses indicated a dramatic shift in the dominant bacterial communities from single-material composting to co-composting. Notably, compared with PM, PM + CM increased the quantity of xylan-degrading bacteria and reduced the quantity of human pathogens. © 2014 The Authors. Microbial Biotechnology published by John Wiley & Sons Ltd and Society for Applied Microbiology.

  2. Comparison of bacterial community structure and dynamics during the thermophilic composting of different types of solid wastes: anaerobic digestion residue, pig manure and chicken manure

    PubMed Central

    Song, Caihong; Li, Mingxiao; Jia, Xuan; Wei, Zimin; Zhao, Yue; Xi, Beidou; Zhu, Chaowei; Liu, Dongming

    2014-01-01

    This study investigated the impact of composting substrate types on the bacterial community structure and dynamics during composting processes. To this end, pig manure (PM), chicken manure (CM), a mixture of PM and CM (PM + CM), and a mixture of PM, CM and anaerobic digestion residue (ADR) (PM + CM + ADR) were selected for thermophilic composting. The bacterial community structure and dynamics during the composting process were detected and analysed by polymerase chain reaction–denaturing gradient gel electrophoresis (DGGE) coupled with a statistic analysis. The physical-chemical analyses indicated that compared to single-material composting (PM, CM), co-composting (PM + CM, PM + CM + ADR) could promote the degradation of organic matter and strengthen the ability of conserving nitrogen. A DGGE profile and statistical analysis demonstrated that co-composting, especially PM + CM + ADR, could improve the bacterial community structure and functional diversity, even in the thermophilic stage. Therefore, co-composting could weaken the screening effect of high temperature on bacterial communities. Dominant sequencing analyses indicated a dramatic shift in the dominant bacterial communities from single-material composting to co-composting. Notably, compared with PM, PM + CM increased the quantity of xylan-degrading bacteria and reduced the quantity of human pathogens. PMID:24963997

  3. Changes in microbial community during hydrogen and methane production in two-stage thermophilic anaerobic co-digestion process from biowaste.

    PubMed

    Zahedi, S; Solera, R; Micolucci, F; Cavinato, C; Bolzonella, D

    2016-03-01

    In this paper, the microbial community in a two-phase thermophilic anaerobic co-digestion process was investigated for its role in hydrogen and methane production, treating waste activated sludge and treating the organic fraction of municipal solid waste. In the acidogenic phase, in which hydrogen is produced, Clostridium sp. clusters represented 76% of total Firmicutes. When feeding the acidogenic effluent into the methanogenic reactors, these acidic conditions negatively influenced methanogenic microorganisms: Methanosaeta sp., (Methanobacteriales, Methanomicrobiales, Methanococcales) decreased by 75%, 50%, 38% and 52%, respectively. At the same time, methanogenic digestion lowered the numbers of Clostridium sp. clusters due to both pH increasing and substrate reduction, and an increase in both Firmicutes genera (non Clostridium) and methanogenic microorganisms, especially Methanosaeta sp. (208%). This was in accordance with the observed decrease in acetic (98%) and butyric (100%) acid contents. To ensure the activity of the acetate-utilizing methanogens (AUM) and the acetogens, high ratios of H2-utilizing methanogens (HUM)/AUM (3.6) were required. Copyright © 2016 Elsevier Ltd. All rights reserved.

  4. Biokinetics and bacterial communities of propionate oxidizing bacteria in phased anaerobic sludge digestion systems.

    PubMed

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

    2013-03-15

    Phased anaerobic digestion is a promising technology and may be a potential source of bio-energy production. Anaerobic digesters are widely used for sewage sludge stabilization and thus a better understanding of the microbial process and kinetics may allow increased volatile solids reduction and methane production through robust process operation. In this study, we analyzed the impact of phase separation and operational conditions on the bio-kinetic characteristics and communities of bacteria associated with four phased anaerobic digestion systems. In addition to significant differences between bacterial communities associated with different digester operating temperatures, our results also revealed that bacterial communities in the phased anaerobic digestion systems differed between the 1st and 2nd phase digesters and we identified strong community composition correlations with several measured physicochemical parameters. The maximum specific growth rates of propionate oxidizing bacteria (POB) in the mesophilic and thermophilic 1st phases were 11 and 23.7 mgCOD mgCOD(-1) d(-1), respectively, while those of the mesophilic and thermophilic 2nd-phase digesters were 6.7 and 18.6 mgCOD mgCOD(-1) d(-1), respectively. Hence, the biokinetic characteristics of the POB population were dependent on the digester loading. In addition, we observed that the temperature dependency factor (θ) values were higher for the less heavily loaded digesters as compared to the values obtained for the 1st-phase digesters. Our results suggested the appropriate application of two sets of POB bio-kinetic that reflect the differing growth responses as a function of propionate concentration (and/or organic loading rates). Also, modeling acetogenesis in phased anaerobic sludge digestion systems will be improved considering a population shift in separate phases. On the basis of the bio-kinetic values estimated in various digesters, high levels of propionate in the thermophilic digesters may be

  5. Sulfate addition as an effective method to improve methane fermentation performance and propionate degradation in thermophilic anaerobic co-digestion of coffee grounds, milk and waste activated sludge with AnMBR.

    PubMed

    Li, Qian; Li, Yu-You; Qiao, Wei; Wang, Xiaochang; Takayanagi, Kazuyuki

    2015-06-01

    This study was conducted to investigate the effects of sulfate on propionate degradation and higher organic loading rate (OLR) achievement in a thermophilic AnMBR for 373days using coffee grounds, milk and waste activated sludge (WAS) as the co-substrate. Without the addition of sulfate, the anaerobic system failed at an OLR of 14.6g-COD/L/d, with propionate accumulating to above 2.23g-COD/L, and recovery by an alkalinity supplement was not successful. After sulfate was added into substrates at a COD/SO4(2-) ratio of 200:1 to 350:1, biogas production increased proportionally with OLR increasing from 4.06 to 15.2g-COD/L/d. Propionic acid was maintained at less than 100mg-COD/L due to the effective conversion of propionic acid to methane after the sulfate supplement was added. The long-term stable performance of the AnMBR indicated that adding sulfate was beneficial for the degradation of propionate and achieving a higher OLR under the thermophilic condition. Copyright © 2015 Elsevier Ltd. All rights reserved.

  6. Dry-thermophilic anaerobic digestion of organic fraction of municipal solid waste: methane production modeling.

    PubMed

    Fdez-Güelfo, L A; Alvarez-Gallego, C; Sales, D; García, L I Romero

    2012-03-01

    The influence of particle size and organic matter content of organic fraction of municipal solid waste (OFMSW) in the overall kinetics of dry (30% total solids) thermophilic (55°C) anaerobic digestion have been studied in a semi-continuous stirred tank reactor (SSTR). Two types of wastes were used: synthetic OFMSW (average particle size of 1mm; 0.71 g Volatile Solids/g waste), and OFMSW coming from a composting full scale plant (average particle size of 30 mm; 0.16 g Volatile Solids/g waste). A modification of a widely-validated product-generation kinetic model has been proposed. Results obtained from the modified-model parameterization at steady-state (that include new kinetic parameters as K, Y(pMAX) and θ(MIN)) indicate that the features of the feedstock strongly influence the kinetics of the process. The overall specific growth rate of microorganisms (μ(max)) with synthetic OFMSW is 43% higher compared to OFMSW coming from a composting full scale plant: 0.238 d(-1) (K=1.391 d(-1); Y(pMAX)=1.167 L CH(4)/gDOC(c); θ(MIN)=7.924 days) vs. 0.135 d(-1) (K=1.282 d(-1); Y(pMAX)=1.150 L CH(4)/gDOC(c); θ(MIN)=9.997 days) respectively. Finally, it could be emphasized that the validation of proposed modified-model has been performed successfully by means of the simulation of non-steady state data for the different SRTs tested with each waste. Copyright © 2011 Elsevier Ltd. All rights reserved.

  7. Population dynamics during startup of thermophilic anaerobic digesters: the mixing factor.

    PubMed

    Ghanimeh, Sophia A; Saikaly, Pascal E; Li, Dong; El-Fadel, Mutasem

    2013-11-01

    Two thermophilic digesters were inoculated with manure and started-up under mixed and stagnant conditions. The Archaea in the mixed digester (A) were dominated by hydrogenotrophic Methanobateriaceae (61%) with most of the methane being produced via syntrophic pathways. Methanosarcinales (35%) were the only acetoclastic methanogens present. Acetate dissipation seems to depend on balanced hydrogenotrophic-to-acetotrophic abundance, which in turn was statistically correlated to free ammonia levels. Relative abundance of bacterial community was associated with the loading rate. However, in the absence of mixing (digester B), the relationship between microbial composition and operating parameters was not discernible. This was attributed to the development of microenvironments where environmental conditions are significantly different from average measured parameters. The impact of microenvironments was accentuated by the use of a non-acclimated seed that lacks adequate propionate degraders. Failure to disperse the accumulated propionate, and other organics, created high concentration niches where competitive and inhibiting conditions developed and favored undesired genera, such as Halobacteria (65% in B). As a result, digester B experienced higher acid levels and lower allowable loading rate. Mixing was found necessary to dissipate potential inhibitors, and improve stability and loading capacity, particularly when a non-acclimated seed, often lacking balanced thermophilic microflora, is used. Copyright © 2013 Elsevier Ltd. All rights reserved.

  8. Mesophilic and thermophilic anaerobic co-digestion of rendering plant and slaughterhouse wastes.

    PubMed

    Bayr, Suvi; Rantanen, Marianne; Kaparaju, Prasad; Rintala, Jukka

    2012-01-01

    Co-digestion of rendering and slaughterhouse wastes was studied in laboratory scale semi-continuously fed continuously stirred tank reactors (CSTRs) at 35 and 55 °C. All in all, 10 different rendering plant and slaughterhouse waste fractions were characterised showing high contents of lipids and proteins, and methane potentials of 262-572 dm(3)CH(4)/kg volatile solids(VS)(added). In mesophilic CSTR methane yields of ca 720 dm(3) CH(4)/kg VS(fed) were obtained with organic loading rates (OLR) of 1.0 and 1.5 kg VS/m(3) d, and hydraulic retention time (HRT) of 50 d. For thermophilic process, the lowest studied OLR of 1.5 kg VS/m(3) d, turned to be unstable after operation of 1.5 HRT, due to accumulating ammonia, volatile fatty acids (VFAs) and probably also long chain fatty acids (LCFAs). In conclusion, mesophilic process was found to be more feasible for co-digestion than thermophilic process, methane yields being higher and process more stable in mesophilic conditions. Copyright © 2011 Elsevier Ltd. All rights reserved.

  9. Role of Mn2+ and compatible solutes in the radiation resistance of thermophilic bacteria and archaea.

    PubMed

    Webb, Kimberly M; DiRuggiero, Jocelyne

    2012-01-01

    Radiation-resistant bacteria have garnered a great deal of attention from scientists seeking to expose the mechanisms underlying their incredible survival abilities. Recent analyses showed that the resistance to ionizing radiation (IR) in the archaeon Halobacterium salinarum is dependent upon Mn-antioxidant complexes responsible for the scavenging of reactive oxygen species (ROS) generated by radiation. Here we examined the role of the compatible solutes trehalose, mannosylglycerate, and di-myo-inositol phosphate in the radiation resistance of aerobic and anaerobic thermophiles. We found that the IR resistance of the thermophilic bacteria Rubrobacter xylanophilus and Rubrobacter radiotolerans was highly correlated to the accumulation of high intracellular concentration of trehalose in association with Mn, supporting the model of Mn(2+)-dependent ROS scavenging in the aerobes. In contrast, the hyperthermophilic archaea Thermococcus gammatolerans and Pyrococcus furiosus did not contain significant amounts of intracellular Mn, and we found no significant antioxidant activity from mannosylglycerate and di-myo-inositol phosphate in vitro. We therefore propose that the low levels of IR-generated ROS under anaerobic conditions combined with highly constitutively expressed detoxification systems in these anaerobes are key to their radiation resistance and circumvent the need for the accumulation of Mn-antioxidant complexes in the cell.

  10. 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. Copyright © 2012 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.

  11. PCR detection of thermophilic spore-forming bacteria involved in canned food spoilage.

    PubMed

    Prevost, S; Andre, S; Remize, F

    2010-12-01

    Thermophilic bacteria that form highly heat-resistant spores constitute an important group of spoilage bacteria of low-acid canned food. A PCR assay was developed in order to rapidly trace these bacteria. Three PCR primer pairs were designed from rRNA gene sequences. These primers were evaluated for the specificity and the sensitivity of detection. Two primer pairs allowed detection at the species level of Geobacillus stearothermophilus and Moorella thermoacetica/thermoautrophica. The other pair allowed group-specific detection of anaerobic thermophilic bacteria of the genera Thermoanaerobacterium, Thermoanaerobacter, Caldanerobium and Caldanaerobacter. After a single enrichment step, these PCR assays allowed the detection of 28 thermophiles from 34 cans of spoiled low-acid food. In addition, 13 ingredients were screened for the presence of these bacteria. This PCR assay serves as a detection method for strains able to spoil low-acid canned food treated at 55°C. It will lead to better reactivity in the canning industry. Raw materials and ingredients might be qualified not only for quantitative spore contamination, but also for qualitative contamination by highly heat-resistant spores.

  12. Comparison of mesophilic and thermophilic anaerobic digestion of sugar beet pulp: performance, dewaterability and foam control.

    PubMed

    Suhartini, Sri; Heaven, Sonia; Banks, Charles J

    2014-01-01

    Digestion of sugar beet pulp was assessed in relation to biogas and methane production, foaming potential, and digestate dewaterability. Four 4-litre working volume digesters were operated mesophilically (37±0.5 °C) and four thermophilically (55±0.5 °C) over three hydraulic retention times. Digesters were operated in duplicate at organic loading rates (OLR) of 4 and 5 g volatile solids l(-1) day(-1) without water addition. Thermophilic digestion gave higher biogas and methane productivity than mesophilic and was able to operate at the higher OLR, where mesophilic digestion showed signs of instability. Digestate dewaterability was assessed using capillary suction time and frozen image centrifugation. The occurrence of, or potential for, stable foam formation was assessed using a foaming potential test. Thermophilic operation allowed higher loadings to be applied without loss of performance, and gave a digestate with superior dewatering characteristics and very little foaming potential. Copyright © 2013 Elsevier Ltd. All rights reserved.

  13. Diversity of thermophilic populations during thermophilic aerobic digestion of potato peel slurry.

    PubMed

    Ugwuanyi, J O; Harvey, L M; McNeil, B

    2008-01-01

    To study the diversity of thermophiles during thermophilic aerobic digestion (TAD) of agro-food waste slurries under conditions similar to full-scale processes. Population diversity and development in TAD were studied by standard microbiological techniques and the processes monitored by standard fermentation procedures. Facultative thermophiles were identified as Bacillus coagulans and B. licheniformis, while obligate thermophiles were identified as B. stearothermophilus. They developed rapidly to peaks of 10(7) to 10(8) in thermophiles increased with process temperatures. Thermophiles were unstable at process pH above or below neutral, but developed rapidly at all aeration rates. Peak populations were higher in the median than at extremes of aeration rates. Obligate thermophiles were unstable at low aeration rates. Process self-heating was higher at lower than at higher aeration rate. Beyond 96 h most thermophiles were present as spores. Limited range of indigenous thermophiles drives TAD of slurry. They develop rapidly and are stable at most digestion conditions. Development and stability of thermophiles in TAD suggest that the process may be operated in a wide range of conditions; and even at short HRT in continuous processes without compromising efficiency.

  14. Microbial community structure associated with the high loading anaerobic codigestion of olive mill and abattoir wastewaters.

    PubMed

    Gannoun, Hana; Omri, Ilhem; Chouari, Rakia; Khelifi, Eltaief; Keskes, Sajiaa; Godon, Jean-Jacques; Hamdi, Moktar; Sghir, Abdelghani; Bouallagui, Hassib

    2016-02-01

    The effect of increasing the organic loading rates (OLRs) on the performance of the anaerobic codigestion of olive mill (OMW) and abattoir wastewaters (AW) was investigated under mesophilic and thermophilic conditions. The structure of the microbial community was also monitored. Increasing OLR to 9g of chemical oxygen demand (COD) L(-1)d(-1) affected significantly the biogas yield and microbial diversity at 35°C. However, at 55°C digester remained stable until OLR of 12g of CODL(-1)d(-1) with higher COD removal (80%) and biogas yield (0.52Lg(-1) COD removed). Significant differences in the bacterial communities were detected between mesophilic and thermophilic conditions. The dominant phyla detected in the digester at both phases were the Firmicutes, Actinobacteria, Bacteroidetes, Synergistetes and Spirochaete. However, Verrucomicrobia, Proteobacteria and the candidate division BRC1 were only detected at thermophilic conditions. The Methanobacteriales and the Thermoplasmales were found as a high predominant archaeal member in the anaerobic sludge. Copyright © 2015 Elsevier Ltd. All rights reserved.

  15. Comparing mesophilic and thermophilic anaerobic digestion of chicken manure: Microbial community dynamics and process resilience

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Niu, Qigui; Takemura, Yasuyuki; Kubota, Kengo

    Highlights: • Microbial community dynamics and process functional resilience were investigated. • The threshold of TAN in mesophilic reactor was higher than the thermophilic reactor. • The recoverable archaeal community dynamic sustained the process resilience. • Methanosarcina was more sensitive than Methanoculleus on ammonia inhibition. • TAN and FA effects the dynamic of hydrolytic and acidogenic bacteria obviously. - Abstract: While methane fermentation is considered as the most successful bioenergy treatment for chicken manure, the relationship between operational performance and the dynamic transition of archaeal and bacterial communities remains poorly understood. Two continuous stirred-tank reactors were investigated under thermophilic andmore » mesophilic conditions feeding with 10%TS. The tolerance of thermophilic reactor on total ammonia nitrogen (TAN) was found to be 8000 mg/L with free ammonia (FA) 2000 mg/L compared to 16,000 mg/L (FA1500 mg/L) of mesophilic reactor. Biomethane production was 0.29 L/gV S{sub in} in the steady stage and decreased following TAN increase. After serious inhibition, the mesophilic reactor was recovered successfully by dilution and washing stratagem compared to the unrecoverable of thermophilic reactor. The relationship between the microbial community structure, the bioreactor performance and inhibitors such as TAN, FA, and volatile fatty acid was evaluated by canonical correspondence analysis. The performance of methanogenic activity and substrate removal efficiency were changed significantly correlating with the community evenness and phylogenetic structure. The resilient archaeal community was found even after serious inhibition in both reactors. Obvious dynamics of bacterial communities were observed in acidogenic and hydrolytic functional bacteria following TAN variation in the different stages.« less

  16. Extremely thermophilic microorganisms as metabolic engineering platforms for production of fuels and industrial chemicals.

    PubMed

    Zeldes, Benjamin M; Keller, Matthew W; Loder, Andrew J; Straub, Christopher T; Adams, Michael W W; Kelly, Robert M

    2015-01-01

    Enzymes from extremely thermophilic microorganisms have been of technological interest for some time because of their ability to catalyze reactions of industrial significance at elevated temperatures. Thermophilic enzymes are now routinely produced in recombinant mesophilic hosts for use as discrete biocatalysts. Genome and metagenome sequence data for extreme thermophiles provide useful information for putative biocatalysts for a wide range of biotransformations, albeit involving at most a few enzymatic steps. However, in the past several years, unprecedented progress has been made in establishing molecular genetics tools for extreme thermophiles to the point that the use of these microorganisms as metabolic engineering platforms has become possible. While in its early days, complex metabolic pathways have been altered or engineered into recombinant extreme thermophiles, such that the production of fuels and chemicals at elevated temperatures has become possible. Not only does this expand the thermal range for industrial biotechnology, it also potentially provides biodiverse options for specific biotransformations unique to these microorganisms. The list of extreme thermophiles growing optimally between 70 and 100°C with genetic toolkits currently available includes archaea and bacteria, aerobes and anaerobes, coming from genera such as Caldicellulosiruptor, Sulfolobus, Thermotoga, Thermococcus, and Pyrococcus. These organisms exhibit unusual and potentially useful native metabolic capabilities, including cellulose degradation, metal solubilization, and RuBisCO-free carbon fixation. Those looking to design a thermal bioprocess now have a host of potential candidates to choose from, each with its own advantages and challenges that will influence its appropriateness for specific applications. Here, the issues and opportunities for extremely thermophilic metabolic engineering platforms are considered with an eye toward potential technological advantages for high

  17. Extremely thermophilic microorganisms as metabolic engineering platforms for production of fuels and industrial chemicals

    PubMed Central

    Zeldes, Benjamin M.; Keller, Matthew W.; Loder, Andrew J.; Straub, Christopher T.; Adams, Michael W. W.; Kelly, Robert M.

    2015-01-01

    Enzymes from extremely thermophilic microorganisms have been of technological interest for some time because of their ability to catalyze reactions of industrial significance at elevated temperatures. Thermophilic enzymes are now routinely produced in recombinant mesophilic hosts for use as discrete biocatalysts. Genome and metagenome sequence data for extreme thermophiles provide useful information for putative biocatalysts for a wide range of biotransformations, albeit involving at most a few enzymatic steps. However, in the past several years, unprecedented progress has been made in establishing molecular genetics tools for extreme thermophiles to the point that the use of these microorganisms as metabolic engineering platforms has become possible. While in its early days, complex metabolic pathways have been altered or engineered into recombinant extreme thermophiles, such that the production of fuels and chemicals at elevated temperatures has become possible. Not only does this expand the thermal range for industrial biotechnology, it also potentially provides biodiverse options for specific biotransformations unique to these microorganisms. The list of extreme thermophiles growing optimally between 70 and 100°C with genetic toolkits currently available includes archaea and bacteria, aerobes and anaerobes, coming from genera such as Caldicellulosiruptor, Sulfolobus, Thermotoga, Thermococcus, and Pyrococcus. These organisms exhibit unusual and potentially useful native metabolic capabilities, including cellulose degradation, metal solubilization, and RuBisCO-free carbon fixation. Those looking to design a thermal bioprocess now have a host of potential candidates to choose from, each with its own advantages and challenges that will influence its appropriateness for specific applications. Here, the issues and opportunities for extremely thermophilic metabolic engineering platforms are considered with an eye toward potential technological advantages for high

  18. Evaluation of pretreatment methods on mixed inoculum for both batch and continuous thermophilic biohydrogen production from cassava stillage.

    PubMed

    Luo, Gang; Xie, Li; Zou, Zhonghai; Wang, Wen; Zhou, Qi

    2010-02-01

    Anaerobic sludges, pretreated by chloroform, base, acid, heat and loading-shock, as well as untreated sludge were evaluated for their thermophilic fermentative hydrogen-producing characters from cassava stillage in both batch and continuous experiments. Results showed that the highest hydrogen production was obtained by untreated sludge and there were significant differences (p<0.05) in hydrogen yields (varied from 32.9 to 65.3mlH(2)/gVS) among the tested pretreatment methods in batch experiments. However, the differences in hydrogen yields disappeared in continuous experiments, which indicated the pretreatment methods had only short-term effects on the hydrogen production. Further study showed that alkalinity was a crucial parameter influencing the fermentation process. When the influent was adjusted to pH 6 by NaHCO(3) instead of NaOH, the hydrogen yield increased from about 40 to 52mlH(2)/gVS in all the experiments. Therefore, pretreatment of anaerobic sludge is unnecessary for practical thermophilic fermentative hydrogen production from cassava stillage.

  19. Thermophilic treatment of acidified and partially acidified wastewater using an anaerobic submerged MBR: Factors affecting long-term operational flux.

    PubMed

    Jeison, D; van Lier, J B

    2007-09-01

    The long-term operation of two thermophilic anaerobic submerged membrane bioreactors (AnSMBRs) was studied using acidified and partially acidified synthetic wastewaters. In both reactors, cake formation was identified as the key factor governing critical flux. Even though cake formation was observed to be mostly reversible, particle deposition proceeds fast once the critical flux is exceeded. Very little irreversible fouling was observed during long-term operation, irrespective of the substrate. Critical flux values at the end of the reactors operation were 7 and 3L/m(2)h for the AnSMBRs fed with acidified and partially acidified wastewaters, respectively, at a gas superficial velocity of 70m/h. Small particle size was identified as the responsible parameter for the low observed critical flux values. The degree of wastewater acidification significantly affected the physical properties of the sludge, determining the attainable flux. Based on the fluxes observed in this research, the membrane costs would be in the range of 0.5euro/m(3) of treated wastewater. Gas sparging was ineffective in increasing the critical flux values. However, preliminary tests showed that cross-flow operation may be a feasible alternative to reduce particle deposition.

  20. Anaerobic carboxydotrophic bacteria in geothermal springs identified using stable isotope probing.

    PubMed

    Brady, Allyson L; Sharp, Christine E; Grasby, Stephen E; Dunfield, Peter F

    2015-01-01

    Carbon monoxide (CO) is a potential energy and carbon source for thermophilic bacteria in geothermal environments. Geothermal sites ranging in temperature from 45 to 65°C were investigated for the presence and activity of anaerobic CO-oxidizing bacteria. Anaerobic CO oxidation potentials were measured at up to 48.9 μmoles CO g(-1) (wet weight) day(-1) within five selected sites. Active anaerobic carboxydotrophic bacteria were identified using (13)CO DNA stable isotope probing (SIP) combined with pyrosequencing of 16S rRNA genes amplified from labeled DNA. Bacterial communities identified in heavy DNA fractions were predominated by Firmicutes, which comprised up to 95% of all sequences in (13)CO incubations. The predominant bacteria that assimilated (13)C derived from CO were closely related (>98% 16S rRNA gene sequence identity) to genera of known carboxydotrophs including Thermincola, Desulfotomaculum, Thermolithobacter, and Carboxydocella, although a few species with lower similarity to known bacteria were also found that may represent previously unconfirmed CO-oxidizers. While the distribution was variable, many of the same OTUs were identified across sample sites from different temperature regimes. These results show that bacteria capable of using CO as a carbon source are common in geothermal springs, and that thermophilic carboxydotrophs are probably already quite well known from cultivation studies.

  1. Development of anaerobic digestion methods for palm oil mill effluent (POME) treatment.

    PubMed

    Poh, P E; Chong, M F

    2009-01-01

    Palm oil mill effluent (POME) is a highly polluting wastewater that pollutes the environment if discharged directly due to its high chemical oxygen demand (COD) and biochemical oxygen demand (BOD) concentration. Anaerobic digestion has been widely used for POME treatment with large emphasis placed on capturing the methane gas released as a product of this biodegradation treatment method. The anaerobic digestion method is recognized as a clean development mechanism (CDM) under the Kyoto protocol. Certified emission reduction (CER) can be obtained by using methane gas as a renewable energy. This review aims to discuss the various anaerobic treatments of POME and factors that influence the operation of anaerobic treatment. The POME treatment at both mesophilic and thermophilic temperature ranges are also analyzed.

  2. Biohydrogen production from arabinose and glucose using extreme thermophilic anaerobic mixed cultures

    PubMed Central

    2012-01-01

    Background Second generation hydrogen fermentation technologies using organic agricultural and forestry wastes are emerging. The efficient microbial fermentation of hexoses and pentoses resulting from the pretreatment of lingocellulosic materials is essential for the success of these processes. Results Conversion of arabinose and glucose to hydrogen, by extreme thermophilic, anaerobic, mixed cultures was studied in continuous (70°C, pH 5.5) and batch (70°C, pH 5.5 and pH 7) assays. Two expanded granular sludge bed (EGSB) reactors, Rarab and Rgluc, were continuously fed with arabinose and glucose, respectively. No significant differences in reactor performance were observed for arabinose and glucose organic loading rates (OLR) ranging from 4.3 to 7.1 kgCOD m-3 d-1. However, for an OLR of 14.2 kgCOD m-3 d-1, hydrogen production rate and hydrogen yield were higher in Rarab than in Rgluc (average hydrogen production rate of 3.2 and 2.0 LH2 L-1 d-1 and hydrogen yield of 1.10 and 0.75 molH2 mol-1substrate for Rarab and Rgluc, respectively). Lower hydrogen production in Rgluc was associated with higher lactate production. Denaturing gradient gel electrophoresis (DGGE) results revealed no significant difference on the bacterial community composition between operational periods and between the reactors. Increased hydrogen production was observed in batch experiments when hydrogen partial pressure was kept low, both with arabinose and glucose as substrate. Sugars were completely consumed and hydrogen production stimulated (62% higher) when pH 7 was used instead of pH 5.5. Conclusions Continuous hydrogen production rate from arabinose was significantly higher than from glucose, when higher organic loading rate was used. The effect of hydrogen partial pressure on hydrogen production from glucose in batch mode was related to the extent of sugar utilization and not to the efficiency of substrate conversion to hydrogen. Furthermore, at pH 7.0, sugars uptake, hydrogen production

  3. Defluviitalea phaphyphila sp. nov., a Novel Thermophilic Bacterium That Degrades Brown Algae.

    PubMed

    Ji, Shi-Qi; Wang, Bing; Lu, Ming; Li, Fu-Li

    2016-02-01

    Brown algae are one of the largest groups of oceanic primary producers for CO2 removal and carbon sinks for coastal regions. However, the mechanism for brown alga assimilation remains largely unknown in thermophilic microorganisms. In this work, a thermophilic alginolytic community was enriched from coastal sediment, from which an obligate anaerobic and thermophilic bacterial strain, designated Alg1, was isolated. Alg1 shared a 16S rRNA gene identity of 94.6% with Defluviitalea saccharophila LIND6LT2(T). Phenotypic, chemotaxonomic, and phylogenetic studies suggested strain Alg1 represented a novel species of the genus Defluviitalea, for which the name Defluviitalea phaphyphila sp. nov. is proposed. Alg1 exhibited an intriguing ability to convert carbohydrates of brown algae, including alginate, laminarin, and mannitol, to ethanol and acetic acid. Three gene clusters participating in this process were predicted to be in the genome, and candidate enzymes were successfully expressed, purified, and characterized. Six alginate lyases were demonstrated to synergistically deconstruct alginate into unsaturated monosaccharide, followed by one uronic acid reductase and two 2-keto-3-deoxy-d-gluconate (KDG) kinases to produce pyruvate. A nonclassical mannitol 1-phosphate dehydrogenase, catalyzing D-mannitol 1-phosphate to fructose 1-phosphate in the presence of NAD(+), and one laminarase also were disclosed. This work revealed that a thermophilic brown alga-decomposing system containing numerous novel thermophilic alginate lyases and a unique mannitol 1-phosphate dehydrogenase was adopted by the natural ethanologenic strain Alg1 during the process of evolution in hostile habitats. Copyright © 2016, American Society for Microbiology. All Rights Reserved.

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

    PubMed

    Luo, Gang; Angelidaki, Irini

    2012-11-01

    Biogas produced by anaerobic digestion, is mainly used in a gas motor for heat and electricity production. However, after removal of CO(2) , biogas can be upgraded to natural gas quality, giving more utilization possibilities, such as utilization as autogas, or distant utilization by using the existing natural gas grid. The current study presents a new biological method for biogas upgrading in a separate biogas reactor, containing enriched hydrogenotrophic methanogens and fed with biogas and hydrogen. Both mesophilic- and thermophilic anaerobic cultures were enriched to convert CO(2) to CH(4) by addition of H(2) . Enrichment at thermophilic temperature (55°C) resulted in CO(2) and H(2) bioconversion rate of 320 mL CH(4) /(gVSS h), which was more than 60% higher than that under mesophilic temperature (37°C). Different dominant species were found at mesophilic- and thermophilic-enriched cultures, as revealed by PCR-DGGE. Nonetheless, they all belonged to the order Methanobacteriales, which can mediate hydrogenotrophic methanogenesis. Biogas upgrading was then tested in a thermophilic anaerobic reactor under various operation conditions. By continuous addition of hydrogen in the biogas reactor, high degree of biogas upgrading was achieved. The produced biogas had a CH(4) content, around 95% at steady-state, at gas (mixture of biogas and hydrogen) injection rate of 6 L/(L day). The increase of gas injection rate to 12 L/(L day) resulted in the decrease of CH(4) content to around 90%. Further study showed that by decreasing the gas-liquid mass transfer by increasing the stirring speed of the mixture the CH(4) content was increased to around 95%. Finally, the CH(4) content around 90% was achieved in this study with the gas injection rate as high as 24 L/(L day). Copyright © 2012 Wiley Periodicals, Inc.

  5. Sugar Metabolism of the First Thermophilic Planctomycete Thermogutta terrifontis: Comparative Genomic and Transcriptomic Approaches

    PubMed Central

    Elcheninov, Alexander G.; Menzel, Peter; Gudbergsdottir, Soley R.; Slesarev, Alexei I.; Kadnikov, Vitaly V.; Krogh, Anders; Bonch-Osmolovskaya, Elizaveta A.; Peng, Xu; Kublanov, Ilya V.

    2017-01-01

    Xanthan gum, a complex polysaccharide comprising glucose, mannose and glucuronic acid residues, is involved in numerous biotechnological applications in cosmetics, agriculture, pharmaceuticals, food and petroleum industries. Additionally, its oligosaccharides were shown to possess antimicrobial, antioxidant, and few other properties. Yet, despite its extensive usage, little is known about xanthan gum degradation pathways and mechanisms. Thermogutta terrifontis, isolated from a sample of microbial mat developed in a terrestrial hot spring of Kunashir island (Far-East of Russia), was described as the first thermophilic representative of the Planctomycetes phylum. It grows well on xanthan gum either at aerobic or anaerobic conditions. Genomic analysis unraveled the pathways of oligo- and polysaccharides utilization, as well as the mechanisms of aerobic and anaerobic respiration. The combination of genomic and transcriptomic approaches suggested a novel xanthan gum degradation pathway which involves novel glycosidase(s) of DUF1080 family, hydrolyzing xanthan gum backbone beta-glucosidic linkages and beta-mannosidases instead of xanthan lyases, catalyzing cleavage of terminal beta-mannosidic linkages. Surprisingly, the genes coding DUF1080 proteins were abundant in T. terrifontis and in many other Planctomycetes genomes, which, together with our observation that xanthan gum being a selective substrate for many planctomycetes, suggest crucial role of DUF1080 in xanthan gum degradation. Our findings shed light on the metabolism of the first thermophilic planctomycete, capable to degrade a number of polysaccharides, either aerobically or anaerobically, including the biotechnologically important bacterial polysaccharide xanthan gum. PMID:29163426

  6. Ammonia inhibition on hydrogen enriched anaerobic digestion of manure under mesophilic and thermophilic conditions.

    PubMed

    Wang, Han; Zhang, Yifeng; Angelidaki, Irini

    2016-11-15

    Capturing of carbon dioxide by hydrogen derived from excess renewable energy (e.g., wind mills) to methane in a microbially catalyzed process offers an attractive technology for biogas production and upgrading. This bioconversion process is catalyzed by hydrogenotrophic methanogens, which are known to be sensitive to ammonia. In this study, the tolerance of the biogas process under supply of hydrogen, to ammonia toxicity was studied under mesophilic and thermophilic conditions. When the initial hydrogen partial pressure was 0.5 atm, the methane yield at high ammonia load (7 g NH 4 + -N L -1 ) was 41.0% and 22.3% lower than that at low ammonia load (1 g NH 4 + -N L -1 ) in mesophilic and thermophilic condition, respectively. Meanwhile no significant effect on the biogas composition was observed. Moreover, we found that hydrogentrophic methanogens were more tolerant to the ammonia toxicity than acetoclastic methanogens in the hydrogen enriched biogas production and upgrading processes. The highest methane production yield was achieved under 0.5 atm hydrogen partial pressure in batch reactors at all the tested ammonia levels. Furthermore, the thermophilic methanogens at 0.5 atm of hydrogen partial pressure were more tolerant to high ammonia levels (≥5 g NH 4 + -N L -1 ), compared with mesophilic methanogens. The present study offers insight in developing resistant hydrogen enriched biogas production and upgrading processes treating ammonia-rich waste streams. Copyright © 2016 Elsevier Ltd. All rights reserved.

  7. Thermophilic anaerobic co-digestion of organic fraction of municipal solid waste (OFMSW) with food waste (FW): Enhancement of bio-hydrogen production.

    PubMed

    Angeriz-Campoy, Rubén; Álvarez-Gallego, Carlos J; Romero-García, Luis I

    2015-10-01

    Bio-hydrogen production from dry thermophilic anaerobic co-digestion (55°C and 20% total solids) of organic fraction of municipal solid waste (OFMSW) and food waste (FW) was studied. OFMSW coming from mechanical-biological treatment plants (MBT plants) presents a low organic matter concentration. However, FW has a high organic matter content but several problems by accumulation of volatile fatty acids (VFAs) and system acidification. Tests were conducted using a mixture ratio of 80:20 (OFSMW:FW), to avoid the aforementioned problems. Different solid retention times (SRTs) - 6.6, 4.4, 2.4 and 1.9 days - were tested. It was noted that addition of food waste enhances the hydrogen production in all the SRTs tested. Best results were obtained at 1.9-day SRT. It was observed an increase from 0.64 to 2.51 L H2/L(reactor) day in hydrogen productivity when SRTs decrease from 6.6 to 1.9 days. However, the hydrogen yield increases slightly from 33.7 to 38 mL H2/gVS(added). Copyright © 2015 Elsevier Ltd. All rights reserved.

  8. Continuous Ethanol Fermentation of Pretreated Lignocellulosic Biomasses, Waste Biomasses, Molasses and Syrup Using the Anaerobic, Thermophilic Bacterium Thermoanaerobacter italicus Pentocrobe 411

    PubMed Central

    Andersen, Rasmus Lund; Jensen, Karen Møller; Mikkelsen, Marie Just

    2015-01-01

    Lignocellosic ethanol production is now at a stage where commercial or semi-commercial plants are coming online and, provided cost effective production can be achieved, lignocellulosic ethanol will become an important part of the world bio economy. However, challenges are still to be overcome throughout the process and particularly for the fermentation of the complex sugar mixtures resulting from the hydrolysis of hemicellulose. Here we describe the continuous fermentation of glucose, xylose and arabinose from non-detoxified pretreated wheat straw, birch, corn cob, sugar cane bagasse, cardboard, mixed bio waste, oil palm empty fruit bunch and frond, sugar cane syrup and sugar cane molasses using the anaerobic, thermophilic bacterium Thermoanaerobacter Pentocrobe 411. All fermentations resulted in close to maximum theoretical ethanol yields of 0.47–0.49 g/g (based on glucose, xylose, and arabinose), volumetric ethanol productivities of 1.2–2.7 g/L/h and a total sugar conversion of 90–99% including glucose, xylose and arabinose. The results solidify the potential of Thermoanaerobacter strains as candidates for lignocellulose bioconversion. PMID:26295944

  9. Effects of stabilization and sludge properties in a combined process of anaerobic digestion and thermophilic aerobic digestion.

    PubMed

    Cheng, Jiehong; Kong, Feng; Zhu, Jun; Wu, Xiao

    2015-01-01

    A novel process of combining mesophilic (<35°C) anaerobic digestion with the thermophilic (55°C) aerobic digestion process (AN-TAD) was designed to stabilize sludge and economize aeration energy. Effects of stabilization and sludge properties for AN-TAD process were evaluated by batch experiments during a 25 d digestion period. The sludges digested by AN-TAD process achieved the requirements for Class-A sludge standard. The sludge at total solid (TS) 5.4% had the highest value of decay coefficient K(d(55)) at 0.1851 d(-1) among the three TS contents according to the first-order kinetics equation. Oxidation reduction potential at below 0 mV remained for sludges at TSs of 6.5%, 5.4%, and 4.6% for at least 15 d because of initial hydrolytic-acidification. Concentrations of nitrogen and phosphorus in sludges at TSs of 6.5%, 5.4%, and 4.6% gradually increased up to the highest values in the supernatant during the initial 13 d, causing low utilized value in land application as a fertilizer. Prolonging the retention time for more than 15 d was considered because soluble phosphorus precipitated in the solid phase. High content of soluble organic matters of the soluble chemical oxygen demand, protein, and polysaccharide in the supernatant caused deterioration in sludge dewaterability rates.

  10. Yearlong semi-continuous operation of thermophilic two-stage anaerobic digesters amended with biochar for enhanced biomethane production

    DOE PAGES

    Shen, Yanwen; Forrester, Sara; Koval, Jason; ...

    2017-05-29

    This study aimed to scale up an integrated waste-to-energy system for producing pipelinequality biomethane from shake flasks experiments to two-stage digester systems with semicontinuous operation. The yearlong operation was successfully conducted to compare the performance of thermophilic anaerobic digestion (AD) of sewage sludge amended with corn stover biochar (CSBC) and pine biochar (PBC) to the control under various conditions. Both CSBC and PBC promoted the substrate utilization, methane productivity, and process stability of AD, while CSBC showed superior potential. CSBC enhanced methane content in biogas (CH 4%) and methane production rate (P CHmore » $$_4$$) by up to 25% and 37% respectively in comparison to the control, with maximum CH 4% of 95% and CH 4 yield of 0.34 L/g volatile solid (VS)-added being achieved at steady state. The biochar supplementation also led to a substantial increase of the macro- and micro-nutrients (P, K, Ca, Mg, Fe) by up to 33 times in the digestate, increasing its fertilizer value. Finally, microbial community structure and dynamics were also investigated and compared, and in particular, CSBC promoted the abundance of Clostridia and Methanosarcina. Collectively, this study proves that pyro-biochar as an effective additive material enhances AD performance with continuous operation and that CSBC shows greater potential.« less

  11. Yearlong semi-continuous operation of thermophilic two-stage anaerobic digesters amended with biochar for enhanced biomethane production

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Shen, Yanwen; Forrester, Sara; Koval, Jason

    This study aimed to scale up an integrated waste-to-energy system for producing pipelinequality biomethane from shake flasks experiments to two-stage digester systems with semicontinuous operation. The yearlong operation was successfully conducted to compare the performance of thermophilic anaerobic digestion (AD) of sewage sludge amended with corn stover biochar (CSBC) and pine biochar (PBC) to the control under various conditions. Both CSBC and PBC promoted the substrate utilization, methane productivity, and process stability of AD, while CSBC showed superior potential. CSBC enhanced methane content in biogas (CH 4%) and methane production rate (P CHmore » $$_4$$) by up to 25% and 37% respectively in comparison to the control, with maximum CH 4% of 95% and CH 4 yield of 0.34 L/g volatile solid (VS)-added being achieved at steady state. The biochar supplementation also led to a substantial increase of the macro- and micro-nutrients (P, K, Ca, Mg, Fe) by up to 33 times in the digestate, increasing its fertilizer value. Finally, microbial community structure and dynamics were also investigated and compared, and in particular, CSBC promoted the abundance of Clostridia and Methanosarcina. Collectively, this study proves that pyro-biochar as an effective additive material enhances AD performance with continuous operation and that CSBC shows greater potential.« less

  12. 31P and 13C NMR analyses of the energy metabolism of the thermophilic anaerobe Clostridium thermocellum.

    PubMed

    Tolman, C J; Kanodia, S; Roberts, M F

    1987-08-15

    The energy metabolism of an anaerobic obligate thermophile, Clostridium thermocellum, has been examined as a function of incubation temperature using 31P NMR spectroscopy. Specifically investigated were the generation and availability of ATP as a function of temperature, activation energies for key processes in energy metabolism including formation of a pH gradient across the cell membrane, transport of key nutrients, and initial steps in glycolysis, and the existence of a membrane phase transition in the intact organism. Cells generate ATP via glycolysis at all temperatures examined; hence, limitation of the energy supply is not directly responsible for the lack of growth of this organism at low temperatures. Estimations of activation energies show a distinct hierarchy in the ATP-utilizing reactions examined. Conservation of ATP hydrolysis energy as delta pH has the lowest activation energy (less than or equal to 4 kcal/mol), two transport processes exhibit 10 kcal/mol activation energies, and early phosphorylation steps in glycolysis have significantly higher activation energies (approximately 25 kcal/mol). Neither the membrane-bound ATPase responsible for formation of the pH gradient nor the permease involved in phosphate transport shows evidence of a change in behavior around the phase transition temperature determined for extracted lipids of C. thermocellum. Line widths of inorganic phosphate do show a break in behavior around 35-40 degrees C. Possible explanations for this behavior are discussed.

  13. Thermophilic-anaerobic digestion to produce class A biosolids: initial full-scale studies at Hyperion Treatment Plant.

    PubMed

    Iranpour, R; Cox, H H J; Oh, S; Fan, S; Kearney, R J; Abkian, V; Haug, R T

    2006-02-01

    biosolids are land-applied, require compliance with both bacterial limits. Additional work identified dewatering, cooling of biosolids after the dewatering centrifuges, and contamination as possible factors in the rise in density of fecal coliforms. These results provided the basis for the full conversion of HTP to the Los Angeles continuous-batch, thermophilic-anaerobic-digestion process. During later phases of testing, this process was demonstrated to produce fully disinfected biosolids at the farm for land application.

  14. Stable thermophilic anaerobic digestion of dissolved air flotation (DAF) sludge by co-digestion with swine manure.

    PubMed

    Creamer, K S; Chen, Y; Williams, C M; Cheng, J J

    2010-05-01

    Environmentally sound treatment of by-products in a value-adding process is an ongoing challenge in animal agriculture. The sludge produced as a result of the dissolved air flotation (DAF) wastewater treatment process in swine processing facilities is one such low-value residue. The objective of this study was to determine the fundamental performance parameters for thermophilic anaerobic digestion of DAF sludge. Testing in a semi-continuous stirred tank reactor and in batch reactors was conducted to determine the kinetics of degradation and biogas yield. Stable operation could not be achieved using pure DAF sludge as a substrate, possibly due to inhibition by long-chain fatty acids or to nutrient deficiencies. However, in a 1:1 ratio (w/w, dry basis) with swine manure, operation was both stable and productive. In the semi-continuous stirred reactor at 54.5 degrees Celsius, a hydraulic residence time of 10 days, and an organic loading rate of 4.68 gVS/day/L, the methane production rate was 2.19 L/L/day and the specific methane production rate was 0.47 L/gVS (fed). Maximum specific methanogenic activity (SMA) in batch testing was 0.15 mmoles CH(4) h(-1) gVS(-1) at a substrate concentration of 6.9 gVS L(-1). Higher substrate concentrations cause an initial lag in methane production, possibly due to long-chain fatty acid or nitrogen inhibition. Copyright 2009 Elsevier Ltd. All rights reserved.

  15. Lignocellulose-derived thin stillage composition and efficient biological treatment with a high-rate hybrid anaerobic bioreactor system.

    PubMed

    Oosterkamp, Margreet J; Méndez-García, Celia; Kim, Chang-H; Bauer, Stefan; Ibáñez, Ana B; Zimmerman, Sabrina; Hong, Pei-Ying; Cann, Isaac K; Mackie, Roderick I

    2016-01-01

    This study aims to chemically characterize thin stillage derived from lignocellulosic biomass distillation residues in terms of organic strength, nutrient, and mineral content. The feasibility of performing anaerobic digestion on these stillages at mesophilic (40 °C) and thermophilic (55 °C) temperatures to produce methane was demonstrated. The microbial communities involved were further characterized. Energy and sugar cane stillage have a high chemical oxygen demand (COD of 43 and 30 g/L, respectively) and low pH (pH 4.3). Furthermore, the acetate concentration in sugar cane stillage was high (45 mM) but was not detected in energy cane stillage. There was also a high amount of lactate in both types of stillage (35-37 mM). The amount of sugars was 200 times higher in energy cane stillage compared to sugar cane stillage. Although there was a high concentration of sulfate (18 and 23 mM in sugar and energy cane stillage, respectively), both thin stillages were efficiently digested anaerobically with high COD removal under mesophilic and thermophilic temperature conditions and with an organic loading rate of 15-21 g COD/L/d. The methane production rate was 0.2 L/g COD, with a methane percentage of 60 and 64, and 92 and 94 % soluble COD removed, respectively, by the mesophilic and thermophilic reactors. Although both treatment processes were equally efficient, there were different microbial communities involved possibly arising from the differences in the composition of energy cane and sugar cane stillage. There was more acetic acid in sugar cane stillage which may have promoted the occurrence of aceticlastic methanogens to perform a direct conversion of acetate to methane in reactors treating sugar cane stillage. Results showed that thin stillage contains easily degradable compounds suitable for anaerobic digestion and that hybrid reactors can efficiently convert thin stillage to methane under mesophilic and thermophilic conditions. Furthermore, we found

  16. Mesophilic and thermophilic activated sludge post-treatment of paper mill process water.

    PubMed

    Vogelaar, J C T; Bouwhuis, E; Klapwijk, A; Spanjers, H; van Lier, J B

    2002-04-01

    Increasing system closure in paper mills and higher process water temperatures make the applicability of thermophilic treatment systems increasingly important. The use of activated sludge as a suitable thermophilic post-treatment system for anaerobically pre-treated paper process water from a paper mill using recycled wastepaper was studied. Two lab-scale plug flow activated sludge reactors were run in parallel for 6 months; a thermophilic reactor at 55 degrees C and a reference reactor at 30 degrees C. Both reactors were operated simultaneously at 20, 15 and 10 days SRT. The effects of temperature and SRT on sludge settleability and chemical oxygen demand (COD) removal efficiencies of different fractions were studied. Total COD removal percentages over the whole experimental period were 58+/-5% at 30 degrees C and 48 +/- 10% at 55 degrees C. The effect of the SRT on the total COD removal was negligible. Differences in total COD removal between both systems were due to a lesser removal of soluble and colloidal COD at 55 degrees C compared to the reference system. At 30 degrees C, colloidal COD removal percentages were 65+/-25%, 75+/-17% and 86+/-22% at 20, 15 and 10 days SRT, respectively. At 55 degrees C, these percentages were 48+/-34%, 40+/-28% and 70+/-25%, respectively. The effluent concentrations of colloidal COD in both systems were related to the influent concentration of colloidal material. The thermophilic sludge was not able to retain influent colloidal material as well as the mesophilic sludge causing a higher thermophilic effluent turbidity. Sludge settling properties were excellent in both reactor systems. These were neither temperature nor SRT dependent but were rather caused by extensive calcium precipitation in the aeration tanks creating a very dense sludge. For application in the board industry, a thermophilic in line treatment system seems feasible. The higher effluent turbidity is most likely offset by the energy gains of treatment under

  17. Saponification of fatty slaughterhouse wastes for enhancing anaerobic biodegradability.

    PubMed

    Battimelli, Audrey; Carrère, Hélène; Delgenès, Jean-Philippe

    2009-08-01

    The thermochemical pretreatment by saponification of two kinds of fatty slaughterhouse waste--aeroflotation fats and flesh fats from animal carcasses--was studied in order to improve the waste's anaerobic degradation. The effect of an easily biodegradable compound, ethanol, on raw waste biodegradation was also examined. The aims of the study were to enhance the methanisation of fatty waste and also to show a link between biodegradability and bio-availability. The anaerobic digestion of raw waste, saponified waste and waste with a co-substrate was carried out in batch mode under mesophilic and thermophilic conditions. The results showed little increase in the total volume of biogas, indicating a good biodegradability of the raw wastes. Mean biogas volume reached 1200 mL/g VS which represented more than 90% of the maximal theoretical biogas potential. Raw fatty wastes were slowly biodegraded whereas pretreated wastes showed improved initial reaction kinetics, indicating a better initial bio-availability, particularly for mesophilic runs. The effects observed for raw wastes with ethanol as co-substrate depended on the process temperature: in mesophilic conditions, an initial improvement was observed whereas in thermophilic conditions a significant decrease in biodegradability was observed.

  18. Enhancing ethanol production from thermophilic and mesophilic solid digestate using ozone combined with aqueous ammonia pretreatment.

    PubMed

    Wang, Dianlong; Xi, Jiang; Ai, Ping; Yu, Liang; Zhai, Hong; Yan, Shuiping; Zhang, Yanlin

    2016-05-01

    Pretreatment with ozone combined with aqueous ammonia was used to recover residual organic carbon from recalcitrant solid digestate for ethanol production after anaerobic digestion (AD) of rice straw. Methane yield of AD at mesophilic and thermophilic conditions, and ethanol production of solid digestate were investigated. The results showed that the methane yield at thermophilic temperature was 72.2% higher than that at mesophilic temperature under the same conditions of 24days and 17% solid concentration. And also the ethanol production efficiency of solid digestate after thermophilic process was 24.3% higher than that of solid digestate after mesophilic process. In this study, the optimal conditions for integrated methane and ethanol processes were determined as 55°C, 17% solid concentration and 24days. 58.6% of glucose conversion, 142.8g/kg of methane yield and 65.2g/kg of ethanol yield were achieved, and the highest net energy balance was calculated as 6416kJ/kg. Copyright © 2016 Elsevier Ltd. All rights reserved.

  19. Optimisation of single-phase dry-thermophilic anaerobic digestion under high organic loading rates of industrial municipal solid waste: population dynamics.

    PubMed

    Zahedi, S; Sales, D; Romero, L I; Solera, R

    2013-10-01

    Different high feed organic loading rates (OLRs) (from 5.7 g to 46.0 g TVS/l/d) or hydraulic retention times (HRTs) (from 15 d to 2 d) in single-phase dry-thermophilic anaerobic digestion (AD) of organic fraction municipal solid waste (OFMSW) were investigated. The specific gas production (SGP) values (0.25-0.53 m(3)/kg TVS) and the percentages of Eubacteria, Archaea, H2-utilising methanogens (HUMs) and acetate-utilising methanogens (AUMs) were stable within the ranges 80.2-91.1%, 12.4-18.5%, 4.4-9.8% and 5.5-10.9%, respectively. A HUM/AUM ratio greater than 0.7 seems to be necessary to maintain very low partial pressures of H2 required for dry AD process. Increasing OLR resulted in an increase in all the populations, except for propionate-utilising acetogens (PUAs). Optimal conditions were obtained at 3d HRT (OLR=30.7 g TVS/l/d), which is lower than the doubling time of acetogens and methanogens. The methane production (MP) was clearly higher than those reported in AD of OFMSW. Copyright © 2013 Elsevier Ltd. All rights reserved.

  20. Evaluation of anaerobic digestion processes for short sludge-age waste activated sludge combined with anammox treatment of digestate liquor.

    PubMed

    Ge, Huoqing; Batstone, Damien; Keller, Jurg

    2016-01-01

    The need to reduce energy input and enhance energy recovery from wastewater is driving renewed interest in high-rate activated sludge treatment (i.e. short hydraulic and solids retention times (HRT and SRT, respectively)). This process generates short SRT activated sludge stream, which should be highly degradable. However, the evaluation of anaerobic digestion of short SRT sludge has been limited. This paper assesses anaerobic digestion of short SRT sludge digestion derived from meat processing wastewater under thermophilic and mesophilic conditions. The thermophilic digestion system (55°C) achieved 60 and 68% volatile solids destruction at 8 day and 10 day HRT, respectively, compared with 50% in the mesophilic digestion system (35°C, 10 day HRT). The digestion effluents from the thermophilic (8-10 day HRT) and mesophilic systems were stable, as assessed by residual methane potentials. The ammonia rich sludge dewatering liquor was effectively treated by a batch anammox process, which exhibited comparable nitrogen removal rate as the tests using a control synthetic ammonia solution, indicating that the dewatering liquor did not have inhibiting/toxic effects on the anammox activity.

  1. Anaerobic carboxydotrophic bacteria in geothermal springs identified using stable isotope probing

    PubMed Central

    Brady, Allyson L.; Sharp, Christine E.; Grasby, Stephen E.; Dunfield, Peter F.

    2015-01-01

    Carbon monoxide (CO) is a potential energy and carbon source for thermophilic bacteria in geothermal environments. Geothermal sites ranging in temperature from 45 to 65°C were investigated for the presence and activity of anaerobic CO-oxidizing bacteria. Anaerobic CO oxidation potentials were measured at up to 48.9 μmoles CO g−1 (wet weight) day−1 within five selected sites. Active anaerobic carboxydotrophic bacteria were identified using 13CO DNA stable isotope probing (SIP) combined with pyrosequencing of 16S rRNA genes amplified from labeled DNA. Bacterial communities identified in heavy DNA fractions were predominated by Firmicutes, which comprised up to 95% of all sequences in 13CO incubations. The predominant bacteria that assimilated 13C derived from CO were closely related (>98% 16S rRNA gene sequence identity) to genera of known carboxydotrophs including Thermincola, Desulfotomaculum, Thermolithobacter, and Carboxydocella, although a few species with lower similarity to known bacteria were also found that may represent previously unconfirmed CO-oxidizers. While the distribution was variable, many of the same OTUs were identified across sample sites from different temperature regimes. These results show that bacteria capable of using CO as a carbon source are common in geothermal springs, and that thermophilic carboxydotrophs are probably already quite well known from cultivation studies. PMID:26388850

  2. Biohydrogen production from space crew's waste simulants using thermophilic consolidated bioprocessing.

    PubMed

    Wang, Jia; Bibra, Mohit; Venkateswaran, Kasthuri; Salem, David R; Rathinam, Navanietha Krishnaraj; Gadhamshetty, Venkataraman; Sani, Rajesh K

    2018-05-01

    Human waste simulants were for the first time converted into biohydrogen by a newly developed anaerobic microbial consortium via thermophilic consolidated bioprocessing. Four different BioH 2 -producing consortia (denoted as C1, C2, C3 and C4) were isolated, and developed using human waste simulants as substrate. The thermophilic consortium C3, which contained Thermoanaerobacterium, Caloribacterium, and Caldanaerobius species as the main constituents, showed the highest BioH 2 production (3.999 mmol/g) from human waste simulants under optimized conditions (pH 7.0 and 60 °C). The consortium C3 also produced significant amounts of BioH 2 (5.732 mmol/g and 2.186 mmol/g) using wastewater and activated sludge, respectively. The developed consortium in this study is a promising candidate for H 2 production in space applications as in situ resource utilization. Copyright © 2018 Elsevier Ltd. All rights reserved.

  3. Caldicellulosiruptor kronotskyensis sp. nov. and Caldicellulosiruptor hydrothermalis sp. nov., two extremely thermophilic, cellulolytic, anaerobic bacteria from Kamchatka thermal springs.

    PubMed

    Miroshnichenko, Margarita L; Kublanov, Ilya V; Kostrikina, Nadezhda A; Tourova, Tatyana P; Kolganova, Tatyana V; Birkeland, Nils-Kåre; Bonch-Osmolovskaya, Elizaveta A

    2008-06-01

    Five novel strains (2002(T), 2902, 2006, 108(T) and 117) of cellulose-degrading, anaerobic, thermophilic bacteria were isolated from terrestrial hot springs of Kamchatka (Far East, Russia). Strains 2002(T) and 108(T) were non-spore-forming bacteria with a Gram-positive type cell wall and peritrichous flagella. Optimum growth of strains 2002(T) and 108(T) occurred at pH 7.0 and at temperatures of 70 and 65 degrees C, respectively. The G+C contents of the DNA of strains 2002(T) and 108(T) were 35.1 and 36.4 mol%, respectively. Comparative 16S rRNA gene sequence analysis revealed that the isolates belonged to the genus Caldicellulosiruptor. However, DNA-DNA hybridization experiments indicated that the levels of relatedness between strains 2002(T) and 108(T) and those of recognized members of the genus Caldicellulosiruptor ranged between 32 and 54 %. Based on both phenotypic and genomic differences, strains 2002(T) and 108(T) are considered to represent two novel species of the genus Caldicellulosiruptor. The names proposed for these organisms are Caldicellulosiruptor kronotskyensis sp. nov. (type strain 2002(T)=DSM 18902(T)=VKM B-2412(T)) and Caldicellulosiruptor hydrothermalis sp. nov. (type strain 108(T)=DSM 18901(T)=VKM B-2411(T)).

  4. An extremely thermophilic anaerobic bacterium Caldicellulosiruptor sp. F32 exhibits distinctive properties in growth and xylanases during xylan hydrolysis.

    PubMed

    Ying, Yu; Meng, Dongdong; Chen, Xiaohua; Li, Fuli

    2013-08-15

    An anaerobic, extremely thermophilic, and cellulose- and xylan-degrading bacterium F32 was isolated from biocompost. Sequence analysis of the 16S rRNA gene of this strain showed that it was closely related to Caldicellulosiruptor saccharolyticus DSM 8903 (99.0% identity). Physiological and biochemical data also supported that identification of strain F32 as a Caldicellulosiruptor species. The proteins secreted by Caldicellulosiruptor sp. F32 grown on xylan showed a xylanase activity of 7.74U/mg, which was 2.5 times higher than that of C. saccharolyticus DSM 8903. Based on the genomic sequencing data, 2 xylanase genes, JX030400 and JX030401, were identified in Caldicellulosiruptor sp. F32. The xylanase encoded by JX030401 shared 97% identity with Csac_0696 of C. saccharolyticus DSM 8903, while that encoded by JX030400 shared 94% identity with Athe_0089 of C. bescii DSM 6725, which was not found in the genome of strain DSM 8903. Xylanse encoded by JX030400 had 9-fold higher specific activity than JX030401. Our results indicated that although the 2 strains shared high identity, the xylanase system in Caldicellulosiruptor sp. F32 was more efficient than that in C. saccharolyticus DSM 8903. Copyright © 2013 Elsevier Inc. All rights reserved.

  5. Thermophilic co-digestion feasibility of distillers grains and swine manure: effect of C/N ratio and organic loading rate during high solid anaerobic digestion (HSAD).

    PubMed

    Sensai, P; Thangamani, A; Visvanathan, C

    2014-01-01

    Anaerobic co-digestion of high solids containing distillers grains and swine manure (total solids, 27 +/- 2% and 18 +/- 2%, respectively) was evaluated in this study to assess the effect of C/N ratio and organic loading rate (OLR). Feed mixture was balanced to achieve a C/N ratio of 30/1 by mixing distillers grains and swine manure. Pilot-scale co-digestion of distillers grains and swine manure was carried out under thermophilic conditions in the continuous mode for seven different OLRs from R1 to R7 (3.5, 5, 6, 8, 10, 12 and 14 kg VS/m3 day) under high solid anaerobic digestion. The methane yield and volatile solid (VS) removal were consistent; ranging from 0.33 to 0.34 m3CH4/kg VS day and 50-53%, respectively, until OLR 8 kg VS/m3 day. After which methane yield and VS removal significantly decreased to 0.26 m3 CH4/kg VS day and 42%, respectively, when OLR was increased to 14 kg VS/m3 day. However, during operation, at OLR of 10 kg VS/m3 day, the methane yield and VS removal increased after the 19th day to 0.33 m3 CH4/kg VS day and 46%, respectively, indicating that a longer acclimatization period is required by methanogens at a higher loading rate.

  6. Brockia lithotrophica gen. nov., sp. nov., an anaerobic thermophilic bacterium from a terrestrial hot spring.

    PubMed

    Perevalova, Anna A; Kublanov, Ilya V; Baslerov, R V; Zhang, Gengxin; Bonch-Osmolovskaya, Elizaveta A

    2013-02-01

    A novel thermophilic bacterium, strain Kam1851(T), was isolated from a terrestrial hot spring of the Uzon Caldera, Kamchatka Peninsula, Russia. Cells of strain Kam1851(T) were spore-forming rods with a gram-positive type of cell wall. Growth was observed between 46 and 78 °C, and pH 5.5-8.5. The optimal growth (doubling time, 6.0 h) was at 60-65 °C and pH 6.5. The isolate was an obligate anaerobe growing in pre-reduced medium only. It grew on mineral medium with molecular hydrogen or formate as electron donors, and elemental sulfur, thiosulfate or polysulfide as electron acceptors. The main cellular fatty acids were C(16 : 0) (34.2 %), iso-C(16 : 0) (18 %), C(18 : 0) (12.8 %) and iso-C(17 : 0) (11.1 %). The G+C content of the genomic DNA of strain Kam1851(T) was 63 mol%. 16S rRNA gene sequence analysis showed that strain Kam1851(T) belonged to the order Thermoanaerobacterales, but it was not closely related to representatives of any genera with validly published names. The most closely related strains, which had no more than 89.2 % sequence similarity, were members of the genera Ammonifex and Caldanaerobacter. On the basis of its phylogenetic position and novel phenotypic features, isolate Kam1851(T) is proposed to represent a novel species in a new genus, Brockia lithotrophica gen. nov., sp. nov.; the type strain of Brockia lithotrophica is Kam1851(T) ( = DSM 22653(T) = VKM B-2685(T)).

  7. Thermanaeromonas burensis sp. nov., a thermophilic anaerobe isolated from a subterranean clay environment.

    PubMed

    Gam, Zouhaier Ben Ali; Daumas, Sylvie; Casalot, Laurence; Bartoli-Joseph, Manon; Necib, Sophia; Linard, Yannick; Labat, Marc

    2016-01-01

    A strictly anaerobic, thermophilic and halotolerant strain, designated IA106T, was isolated from the seepage water collected in a metal biocorrosion test at a depth of 490 m, in a 130-160 m thick, subterranean Callovo-Oxfordian clay formation (158-152 million years old) in northern France. This geological formation has been selected as the potential host rock for the French high-level nuclear waste repository. Cells of strain IA106T stained Gram-positive and were non-motile, spore-forming, straight rods (0.5 × 2-6 μm). The five major fatty acids were C16 : 0 (15.9 %), C18 : 0 (15.4 %), iso-C17 : 1 I and/or anteiso-C17 : 1 B(14.8 %), iso-C17 : 0 (14.7 %) and iso-C15 : 0 (13.0 %). Growth was observed at temperatures ranging from 55 to 70 °C and at pH 5.5-9. The salinity range for growth was 0-20 g NaCl 1- 1. Yeast extract was required for growth. Strain IA106T was able to grow on lactate and various sugars in the presence of thiosulfate as electron acceptor. Sulfate, sulfite, elemental sulfur, fumarate, nitrate and nitrite were not reduced. The DNA G+C content was 60.2 mol%. 16S rRNA gene sequence analysis indicated that strain IA106T belonged to the family Thermoanaerobacteraceae, class Clostridia, phylum Firmicutes, and was most closely related to Thermanaeromonas toyohensis DSM 14490T (95.16 % 16S rRNA gene sequence similarity). On the basis of 16S rRNA gene sequence comparisons and physiological characteristics, strain IA106T represents a novel species of the genus Thermanaeromonas, for which the name Thermanaeromonas burensis sp. nov. is proposed. The type strain is IA106T ( = DSM 26576T = JCM 18718T).

  8. Co-digestion of bovine slaughterhouse wastes, cow manure, various crops and municipal solid waste at thermophilic conditions: a comparison with specific case running at mesophilic conditions.

    PubMed

    Pagés-Díaz, J; Sárvári-Horváth, I; Pérez-Olmo, J; Pereda-Reyes, I

    2013-01-01

    A co-digestion process was evaluated when mixing different ratios of agro-industrial residues, i.e. bovine slaughterhouse waste (SB); cow manure (M); various crop residues (VC); and municipal solid waste (MSW) by anaerobic batch digestion under thermophilic conditions (55 °C). A selected study case at mesophilic condition (37 °C) was also investigated. The performance of the co-digestion was evaluated by kinetics (k(0)). The best kinetic results were obtained under thermophilic operation when a mixture of 22% w/w SB, 22% w/w M, 45% w/w VC and 11% w/w MSW was co-digested, which showed a proper combination of high values in r(s)CH(4) and k(0) (0.066 Nm(3)CH(4)/kgVS*d, 0.336 d(-1)) during the anaerobic process. The effect of temperature on methane yield (Y(CH4)), specific methane rate (r(s)CH(4)) and k(0) was also analyzed for a specific study case; there a mixture of 25% w/w of SB, 37.5% w/w of M, 37.5% of VC and 0% of MSW was used. Response variables were severely affected by mesophilic conditions, diminishing to at least 45% of the thermophilic values obtained for a similar mixture. The effect of temperature suggested that thermophilic conditions are suitable to treat these residues.

  9. Electrochemical Performance and Microbial Characterization of Thermophilic Microbial Fuel Cells

    NASA Astrophysics Data System (ADS)

    Wrighton, K. C.; Agbo, P.; Brodie, E. L.; Weber, K. A.; Desantis, T. Z.; Anderson, G. L.; Coates, J. D.

    2007-12-01

    Significant research effort is currently focused on microbial fuel cells (MFC) as a source of renewable energy. To date, most of these efforts have concentrated on MFCs operating at mesophilic temperatures. However, many previous studies have reported on the superiority of thermophilic conditions in anaerobic digestion and demonstrated a net gain in energy yield, in terms of methane, relative to the increased energy requirements of operation. Because of this, our recent studies on MFCs focused on investigating the operation and microbiology associated with thermophilic MFCs operating at 55°C. Over 100-day operation, these MFCs were highly stable and achieved a maximum power density of 24mW/m2 and a columbic efficiency of 89 percent with acetate as the sole electron donor. In order to characterize the microbial community involved in thermophilic electricity generation, DNA and RNA were isolated from the electrode and PhyloChip analyses performed. Exploring the changes in the microbial community over time in electricity producing MFC revealed an increase in relative abundance of populations belonging to the Firmicutes, Chloroflexi, and alpha Proteobacteria by at least one order of magnitude. In contrast, these populations decreased in the open circuit and no electron donor amended controls. In order to better characterize the active microbial populations, we enriched and isolated a novel organism, strain JR, from samples collected from an operating MFC. Based on 16S rRNA sequence analysis strain JR was a member of the family Peptococcaceae, within the Phylum Firmicutes, clustering with Thermincola ferriacetica (98 percent similarity). Phenotypic characterization revealed that strain JR was capable of thermophilic dissimilatory reduction of insoluble electron acceptors such as amorphous Fe(III); as well as reduction of the model quinone 2,6-anthraquinone disulfonate. Thermincola strain JR was also capable of producing current by coupling acetate oxidation to anodic

  10. Methanosarcinaceae and Acetate-Oxidizing Pathways Dominate in High-Rate Thermophilic Anaerobic Digestion of Waste-Activated Sludge

    PubMed Central

    Ho, Dang P.; Jensen, Paul D.

    2013-01-01

    This study investigated the process of high-rate, high-temperature methanogenesis to enable very-high-volume loading during anaerobic digestion of waste-activated sludge. Reducing the hydraulic retention time (HRT) from 15 to 20 days in mesophilic digestion down to 3 days was achievable at a thermophilic temperature (55°C) with stable digester performance and methanogenic activity. A volatile solids (VS) destruction efficiency of 33 to 35% was achieved on waste-activated sludge, comparable to that obtained via mesophilic processes with low organic acid levels (<200 mg/liter chemical oxygen demand [COD]). Methane yield (VS basis) was 150 to 180 liters of CH4/kg of VSadded. According to 16S rRNA pyrotag sequencing and fluorescence in situ hybridization (FISH), the methanogenic community was dominated by members of the Methanosarcinaceae, which have a high level of metabolic capability, including acetoclastic and hydrogenotrophic methanogenesis. Loss of function at an HRT of 2 days was accompanied by a loss of the methanogens, according to pyrotag sequencing. The two acetate conversion pathways, namely, acetoclastic methanogenesis and syntrophic acetate oxidation, were quantified by stable carbon isotope ratio mass spectrometry. The results showed that the majority of methane was generated by nonacetoclastic pathways, both in the reactors and in off-line batch tests, confirming that syntrophic acetate oxidation is a key pathway at elevated temperatures. The proportion of methane due to acetate cleavage increased later in the batch, and it is likely that stable oxidation in the continuous reactor was maintained by application of the consistently low retention time. PMID:23956388

  11. Caldicellulosiruptor obsidiansis sp. nov., an anaerobic, extremely thermophilic, cellulolytic bacterium isolated from Obsidian Pool, Yellowstone National Park.

    PubMed

    Hamilton-Brehm, Scott D; Mosher, Jennifer J; Vishnivetskaya, Tatiana; Podar, Mircea; Carroll, Sue; Allman, Steve; Phelps, Tommy J; Keller, Martin; Elkins, James G

    2010-02-01

    A novel, obligately anaerobic, extremely thermophilic, cellulolytic bacterium, designated OB47(T), was isolated from Obsidian Pool, Yellowstone National Park, WY. The isolate was a nonmotile, non-spore-forming, Gram-positive rod approximately 2 microm long by 0.2 microm wide and grew at temperatures between 55 and 85 degrees C, with the optimum at 78 degrees C. The pH range for growth was 6.0 to 8.0, with values of near 7.0 being optimal. Growth on cellobiose produced the fastest specific growth rate at 0.75 h(-1). The organism also displayed fermentative growth on glucose, maltose, arabinose, fructose, starch, lactose, mannose, sucrose, galactose, xylose, arabinogalactan, Avicel, xylan, filter paper, processed cardboard, pectin, dilute acid-pretreated switchgrass, and Populus. OB47(T) was unable to grow on mannitol, fucose, lignin, Gelrite, acetate, glycerol, ribose, sorbitol, carboxymethylcellulose, and casein. Yeast extract stimulated growth, and thiosulfate, sulfate, nitrate, and sulfur were not reduced. Fermentation end products were mainly acetate, H2, and CO2, although lactate and ethanol were produced in 5-liter batch fermentations. The G+C content of the DNA was 35 mol%, and sequence analysis of the small subunit rRNA gene placed OB47(T) within the genus Caldicellulosiruptor. Based on its phylogenetic and phenotypic properties, the isolate is proposed to be designated Caldicellulosiruptor obsidiansis sp. nov. and OB47 is the type strain (ATCC BAA-2073).

  12. Caldicellulosiruptor obsidiansis sp. nov., an Anaerobic, Extremely Thermophilic, Cellulolytic Bacterium Isolated from Obsidian Pool, Yellowstone National Park▿

    PubMed Central

    Hamilton-Brehm, Scott D.; Mosher, Jennifer J.; Vishnivetskaya, Tatiana; Podar, Mircea; Carroll, Sue; Allman, Steve; Phelps, Tommy J.; Keller, Martin; Elkins, James G.

    2010-01-01

    A novel, obligately anaerobic, extremely thermophilic, cellulolytic bacterium, designated OB47T, was isolated from Obsidian Pool, Yellowstone National Park, WY. The isolate was a nonmotile, non-spore-forming, Gram-positive rod approximately 2 μm long by 0.2 μm wide and grew at temperatures between 55 and 85°C, with the optimum at 78°C. The pH range for growth was 6.0 to 8.0, with values of near 7.0 being optimal. Growth on cellobiose produced the fastest specific growth rate at 0.75 h−1. The organism also displayed fermentative growth on glucose, maltose, arabinose, fructose, starch, lactose, mannose, sucrose, galactose, xylose, arabinogalactan, Avicel, xylan, filter paper, processed cardboard, pectin, dilute acid-pretreated switchgrass, and Populus. OB47T was unable to grow on mannitol, fucose, lignin, Gelrite, acetate, glycerol, ribose, sorbitol, carboxymethylcellulose, and casein. Yeast extract stimulated growth, and thiosulfate, sulfate, nitrate, and sulfur were not reduced. Fermentation end products were mainly acetate, H2, and CO2, although lactate and ethanol were produced in 5-liter batch fermentations. The G+C content of the DNA was 35 mol%, and sequence analysis of the small subunit rRNA gene placed OB47T within the genus Caldicellulosiruptor. Based on its phylogenetic and phenotypic properties, the isolate is proposed to be designated Caldicellulosiruptor obsidiansis sp. nov. and OB47 is the type strain (ATCC BAA-2073). PMID:20023107

  13. Ethanol production efficiency of an anaerobic hemicellulolytic thermophilic bacterium, strain NTOU1, isolated from a marine shallow hydrothermal vent in Taiwan.

    PubMed

    Tsai, Tsai-Ling; Liu, Shiu-Mei; Lee, Shi-Chiang; Chen, Wei-Jei; Chou, Sheng-Hsin; Hsu, Tseng-Chieh; Guo, Gia-Luen; Hwang, Wen-Song; Wiegel, Juergen

    2011-01-01

    A new extremely thermophilic, anaerobic, gram-negative bacterium, strain NTOU1, was enriched and isolated from acidic marine hydrothermal fluids off Gueishandao island in Taiwan with 0.5% starch and 0.5% maltose as carbon sources. This strain was capable of growth utilizing various sugars found in lignocellulosic biomass as well as xylan and cellulose, and produced ethanol, lactate, acetate, and CO(2) as fermentation products. The results of a 16S rRNA gene sequence analysis (1,520 bp) revealed NTOU1 to belong to the genus Thermoanaerobacterium. When tested for the ability to grow and produce ethanol from xylose or rice straw hemicellulosic hydrolysate at 70°C, the strain showed the highest levels of ethanol production (1.65 mol ethanol mol xylose(-1)) in a medium containing 0.5% xylose plus 0.5% yeast extract. Maximum ethanol production from the rice straw hemicellulose was 0.509 g g(-1), equivalent to 98.8% theoretical conversion efficiency. Low concentrations of inhibitors (derived from dilute acid hydrolysis) in the rice straw hemicellulose hydrolysate did not affect the ethanol yield. Thus, Thermoanaerobacterium strain NTOU1 has the potential to be used for ethanol production from hemicellulose.

  14. Hybrid alkali-hydrodynamic disintegration of waste-activated sludge before two-stage anaerobic digestion process.

    PubMed

    Grübel, Klaudiusz; Suschka, Jan

    2015-05-01

    The first step of anaerobic digestion, the hydrolysis, is regarded as the rate-limiting step in the degradation of complex organic compounds, such as waste-activated sludge (WAS). The aim of lab-scale experiments was to pre-hydrolyze the sludge by means of low intensive alkaline sludge conditioning before applying hydrodynamic disintegration, as the pre-treatment procedure. Application of both processes as a hybrid disintegration sludge technology resulted in a higher organic matter release (soluble chemical oxygen demand (SCOD)) to the liquid sludge phase compared with the effects of processes conducted separately. The total SCOD after alkalization at 9 pH (pH in the range of 8.96-9.10, SCOD = 600 mg O2/L) and after hydrodynamic (SCOD = 1450 mg O2/L) disintegration equaled to 2050 mg/L. However, due to the synergistic effect, the obtained SCOD value amounted to 2800 mg/L, which constitutes an additional chemical oxygen demand (COD) dissolution of about 35 %. Similarly, the synergistic effect after alkalization at 10 pH was also obtained. The applied hybrid pre-hydrolysis technology resulted in a disintegration degree of 28-35%. The experiments aimed at selection of the most appropriate procedures in terms of optimal sludge digestion results, including high organic matter degradation (removal) and high biogas production. The analyzed soft hybrid technology influenced the effectiveness of mesophilic/thermophilic anaerobic digestion in a positive way and ensured the sludge minimization. The adopted pre-treatment technology (alkalization + hydrodynamic cavitation) resulted in 22-27% higher biogas production and 13-28% higher biogas yield. After two stages of anaerobic digestion (mesophilic conditions (MAD) + thermophilic anaerobic digestion (TAD)), the highest total solids (TS) reduction amounted to 45.6% and was received for the following sample at 7 days MAD + 17 days TAD. About 7% higher TS reduction was noticed compared with the sample after 9

  15. Novel chemolithotrophic, thermophilic, anaerobic bacteria Thermolithobacter ferrireducens gen. nov., sp. nov. and Thermolithobacter carboxydivorans sp. nov.

    PubMed

    Sokolova, T; Hanel, J; Onyenwoke, R U; Reysenbach, A-L; Banta, A; Geyer, R; González, J M; Whitman, W B; Wiegel, J

    2007-01-01

    Three thermophilic strains of chemolithoautotrophic Fe(III)-reducers were isolated from mixed sediment and water samples (JW/KA-1 and JW/KA-2(T): Calcite Spring, Yellowstone N.P., WY, USA; JW/JH-Fiji-2: Savusavu, Vanu Levu, Fiji). All were Gram stain positive rods (approximately 0.5 x 1.8 microm). Cells occurred singly or in V-shaped pairs, and they formed long chains in complex media. All utilized H(2) to reduce amorphous iron (III) oxide/hydroxide to magnetite at temperatures from 50 to 75 degrees C (opt. approximately 73 degrees C). Growth occurred within the pH(60C) range of 6.5-8.5 (opt. pH(60C) 7.1-7.3). Magnetite production by resting cells occurred at pH(60C) 5.5-10.3 (opt. 7.3). The iron (III) reduction rate was 1.3 mumol Fe(II) produced x h(-1) x ml(-1) in a culture with 3 x 10(7) cells, one of the highest rates reported. In the presence or absence of H(2), JW/KA-2(T) did not utilize CO. The G + C content of the genomic DNA of the type strain is 52.7 +/- 0.3 mol%. Strains JW/KA-1 and JW/KA-2(T) each contain two different 16S rRNA gene sequences. The 16S rRNA gene sequences from JW/KA-1, JW/KA-2(T), or JW/JH-Fiji-2 possessed >99% similarity to each other but also 99% similarity to the 16S rRNA gene sequence from the anaerobic, thermophilic, hydrogenogenic CO-oxidizing bacterium 'Carboxydothermus restrictus' R1. DNA-DNA hybridization between strain JW/KA-2(T) and strain R1(T) yielded 35% similarity. Physiological characteristics and the 16S rRNA gene sequence analysis indicated that the strains represent two novel species and are placed into the novel genus Thermolithobacter within the phylum 'Firmicutes'. In addition, the levels of 16S rRNA gene sequence similarity between the lineage containing the Thermolithobacter and well-established members of the three existing classes of the 'Firmicutes' is less than 85%. Therefore, Thermolithobacter is proposed to constitute the first genus within a novel class of the 'Firmicutes', Thermolithobacteria. The Fe

  16. Long-term stability of thermophilic co-digestion submerged anaerobic membrane reactor encountering high organic loading rate, persistent propionate and detectable hydrogen in biogas.

    PubMed

    Qiao, Wei; Takayanagi, Kazuyuki; Niu, Qigui; Shofie, Mohammad; Li, Yu You

    2013-12-01

    The performance of thermophilic anaerobic co-digestion of coffee grounds and sludge using membrane reactor was investigated for 148 days, out of a total research duration of 263 days. The OLR was increased from 2.2 to 33.7 kg-COD/m(3)d and HRT was shortened from 70 to 7 days. A significant irreversible drop in pH confirmed the overload of reactor. Under a moderately high OLR of 23.6 kg-COD/m(3)d, and with HRT and influent total solids of 10 days and 150 g/L, respectively, the COD removal efficiency was 44.5%. Hydrogen in biogas was around 100-200 ppm, which resulted in the persistent propionate of 1.0-3.2g/L. The VFA consumed approximately 60% of the total alkalinity. NH4HCO3 was supplemented to maintain alkalinity. The stability of system relied on pH management under steady state. The 16SrDNA results showed that hydrogen-utilizing methanogens dominates the archaeal community. The propionate-oxidizing bacteria in bacterial community was insufficient. Copyright © 2013 Elsevier Ltd. All rights reserved.

  17. Development of an efficient anaerobic co-digestion process for garbage, excreta, and septic tank sludge to create a resource recycling-oriented society.

    PubMed

    Sun, Zhao-Yong; Liu, Kai; Tan, Li; Tang, Yue-Qin; Kida, Kenji

    2017-03-01

    In order to develop a resource recycling-oriented society, an efficient anaerobic co-digestion process for garbage, excreta and septic tank sludge was studied based on the quantity of each biomass waste type discharged in Ooki machi, Japan. The anaerobic digestion characteristics of garbage, excreta and 5-fold condensed septic tank sludge (hereafter called condensed sludge) were determined separately. In single-stage mesophilic digestion, the excreta with lower C/N ratios yielded lower biogas volumes and accumulated higher volumes of volatile fatty acid (VFA). On the other hand, garbage allowed for a significantly larger volatile total solid (VTS) digestion efficiency as well as biogas yield by thermophilic digestion. Thus, a two-stage anaerobic co-digestion process consisting of thermophilic liquefaction and mesophilic digestion phases was proposed. In the thermophilic liquefaction of mixed condensed sludge and household garbage (wet mass ratio of 2.2:1), a maximum VTS loading rate of 24g/L/d was achieved. In the mesophilic digestion of mixed liquefied material and excreta (wet mass ratio of 1:1), biogas yield reached approximately 570ml/g-VTS fed with a methane content of 55% at a VTS loading rate of 1.0g/L/d. The performance of the two-stage process was evaluated by comparing it with a single-stage process in which biomass wastes were treated separately. Biogas production by the two-stage process was found to increase by approximately 22.9%. These results demonstrate the effectiveness of a two-stage anaerobic co-digestion process in enhancement of biogas production. Copyright © 2016 Elsevier Ltd. All rights reserved.

  18. Roles of Thermophiles and Fungi in Bitumen Degradation in Mostly Cold Oil Sands Outcrops

    PubMed Central

    Wong, Man-Ling; An, Dongshan; Caffrey, Sean M.; Soh, Jung; Dong, Xiaoli; Sensen, Christoph W.; Oldenburg, Thomas B. P.; Larter, Steve R.

    2015-01-01

    Oil sands are surface exposed in river valley outcrops in northeastern Alberta, where flat slabs (tablets) of weathered, bitumen-saturated sandstone can be retrieved from outcrop cliffs or from riverbeds. Although the average yearly surface temperature of this region is low (0.7°C), we found that the temperatures of the exposed surfaces of outcrop cliffs reached 55 to 60°C on sunny summer days, with daily maxima being 27 to 31°C. Analysis of the cooccurrence of taxa derived from pyrosequencing of 16S/18S rRNA genes indicated that an aerobic microbial network of fungi and hydrocarbon-, methane-, or acetate-oxidizing heterotrophic bacteria was present in all cliff tablets. Metagenomic analyses indicated an elevated presence of fungal cytochrome P450 monooxygenases in these samples. This network was distinct from the heterotrophic community found in riverbeds, which included fewer fungi. A subset of cliff tablets had a network of anaerobic and/or thermophilic taxa, including methanogens, Firmicutes, and Thermotogae, in the center. Long-term aerobic incubation of outcrop samples at 55°C gave a thermophilic microbial community. Analysis of residual bitumen with a Fourier transform ion cyclotron resonance mass spectrometer indicated that aerobic degradation proceeded at 55°C but not at 4°C. Little anaerobic degradation was observed. These results indicate that bitumen degradation on outcrop surfaces is a largely aerobic process with a minor anaerobic contribution and is catalyzed by a consortium of bacteria and fungi. Bitumen degradation is stimulated by periodic high temperatures on outcrop cliffs, which cause significant decreases in bitumen viscosity. PMID:26209669

  19. Unravelling the active microbial community in a thermophilic anaerobic digester-microbial electrolysis cell coupled system under different conditions.

    PubMed

    Cerrillo, Míriam; Viñas, Marc; Bonmatí, August

    2017-03-01

    Thermophilic anaerobic digestion (AD) of pig slurry coupled to a microbial electrolysis cell (MEC) with a recirculation loop was studied at lab-scale as a strategy to increase AD stability when submitted to organic and nitrogen overloads. The system performance was studied, with the recirculation loop both connected and disconnected, in terms of AD methane production, chemical oxygen demand removal (COD) and volatile fatty acid (VFA) concentrations. Furthermore, the microbial population was quantitatively and qualitatively assessed through DNA and RNA-based qPCR and high throughput sequencing (MiSeq), respectively to identify the RNA-based active microbial populations from the total DNA-based microbial community composition both in the AD and MEC reactors under different operational conditions. Suppression of the recirculation loop reduced the AD COD removal efficiency (from 40% to 22%) and the methane production (from 0.32 to 0.03 m 3  m -3  d -1 ). Restoring the recirculation loop led to a methane production of 0.55 m 3  m -3  d -1 concomitant with maximum MEC COD and ammonium removal efficiencies of 29% and 34%, respectively. Regarding microbial analysis, the composition of the AD and MEC anode populations differed from really active microorganisms. Desulfuromonadaceae was revealed as the most active family in the MEC (18%-19% of the RNA relative abundance), while hydrogenotrophic methanogens (Methanobacteriaceae) dominated the AD biomass. Copyright © 2016 Elsevier Ltd. All rights reserved.

  20. Correlation between system performance and bacterial composition under varied mixing intensity in thermophilic anaerobic digestion of food waste.

    PubMed

    Ghanimeh, Sophia A; Al-Sanioura, Dana N; Saikaly, Pascal E; El-Fadel, Mutasem

    2018-01-15

    This study examines the stability and efficiency of thermophilic anaerobic digesters treating food waste under various mixing velocities (50-160 rpm). The results showed that high velocities (120 and 160 rpm) were harmful to the digestion process with 18-30% reduction in methane generation and 1.8 to 3.8 times increase in volatile fatty acids (VFA) concentrations, compared to mild mixing (50 and 80 rpm). Also, the removal rate of soluble COD dropped from 75 to 85% (at 50-80 rpm) to 20-59% (at 120-160 rpm). Similarly, interrupted mixing caused adverse impacts and led to near-failure conditions with excessive VFA accumulation (15.6 g l -1 ), negative removal rate of soluble COD and low methane generation (132 ml gVS -1 ). The best efficiency and stability were achieved under mild mixing (50 and 80 rpm). In particular, the 50 rpm stirring speed resulted in the highest methane generation (573 ml gVS -1 ). High-throughput sequencing of 16S rRNA genes revealed that the digesters were dominated by one bacterial genus (Petrotoga; phylym Thermotogae) at all mixing velocities except at 0 rpm, where the community was dominated by one bacterial genus (Anaerobaculum; phylum Synergistetes). The Petrotoga genus seems to have played a major role in the degradation of organic matter. Copyright © 2017 Elsevier Ltd. All rights reserved.

  1. Isolation and characterization of Keratinibaculum paraultunense gen. nov., sp. nov., a novel thermophilic, anaerobic bacterium with keratinolytic activity.

    PubMed

    Huang, Yan; Sun, Yingjie; Ma, Shichun; Chen, Lu; Zhang, Hui; Deng, Yu

    2013-08-01

    A novel thermophilic, anaerobic, keratinolytic bacterium designated KD-1 was isolated from grassy marshland. Strain KD-1 was a spore-forming rod with a Gram-positive type cell wall, but stained Gram-negative. The temperature, pH, and NaCl concentration range necessary for growth was 30-65 °C (optimum 55 °C), 6.0-10.5 (optimum 8.0-8.5), and 0-6% (optimum 0.2%) (w/v), respectively. Strain KD-1 possessed extracellular keratinase, and the optimum activity of the crude enzyme was pH 8.5 and 70 °C. The enzyme was identified as a thermostable serine-type protease. The strain was sensitive to rifampin, chloramphenicol, kanamycin, and tetracycline and was resistant to erythromycin, neomycin, penicillin, and streptomycin. The main cellular fatty acid was predominantly C15:0 iso (64%), and the G+C content was 28 mol%. Morphological and physiological characterization, together with phylogenetic analysis based on 16S rRNA gene sequencing identified KD-1 as a new species of a novel genus of Clostridiaceae with 95.3%, 93.8% 16S rRNA gene sequence similarity to Clostridium ultunense BS(T) (DSM 10521(T)) and Tepidimicrobium xylanilyticum PML14(T) (= JCM 15035(T)), respectively. We propose the name Keratinibaculum paraultunense gen. nov., sp. nov., with KD-1 (=JCM 18769(T) =DSM 26752(T)) as the type strain. © 2013 Federation of European Microbiological Societies. Published by John Wiley & Sons Ltd. All rights reserved.

  2. An innovative sewage sludge reduction by using a combined mesophilic anaerobic and thermophilic aerobic process with thermal-alkaline treatment and sludge recirculation.

    PubMed

    Cho, Hyun Uk; Park, Sang Kyu; Ha, Jeong Hyub; Park, Jong Moon

    2013-11-15

    Lab-scale High Efficiency Digestion (HED) systems containing a Mesophilic Anaerobic Reactor (MAR), Thermophilic Aerobic Reactor (TAR), liquid/solid separation unit, and thermal-alkaline treatment were developed to evaluate the efficiencies of sludge reduction and methane production. The HED process was divided into three phases to examine the influence of sludge pretreatment and pretreated sludge recirculation using TCOD and VSS reduction, COD solubilization, and methane production. The VSS removal with a solid/liquid separation unit, sludge recirculation, and thermal-alkaline treatment drastically increased up to 95% compared to the feed concentration. In addition, the results of COD solubilization and VSS/TSS showed that the solubilization of cells and organic matters by the thermal-alkaline treatment was highly increased, which was also consistent with the SEM images. In particular, the methane production rate increased 24-fold when the feed sludge and recirculated sludge were pretreated together. Collectively, the HED experiments performed with sludge recirculation and thermal-alkaline treatment demonstrated that the HED systems can be successfully employed for highly efficient sewage sludge reduction and methane gas production. Copyright © 2013 Elsevier Ltd. All rights reserved.

  3. Carboxydobrachium pacificum gen. nov., sp. nov., a new anaerobic, thermophilic, CO-utilizing marine bacterium from Okinawa Trough.

    PubMed

    Sokolova, T G; González, J M; Kostrikina, N A; Chernyh, N A; Tourova, T P; Kato, C; Bonch-Osmolovskaya, E A; Robb, F T

    2001-01-01

    A new anaerobic, thermophilic, CO-utilizing marine bacterium, strain JMT, was isolated from a submarine hot vent in Okinawa Trough. Cells of strain JMT were non-motile thin straight rods, sometimes branching, with a cell wall of the Gram-positive type, surrounded with an S-layer. Chains of three to five cells were often observed. The isolate grew chemolithotrophically on CO, producing equimolar quantities of H2 and CO2 (according to the equation CO+H2O-->CO2+H2) and organotrophically on peptone, yeast extract, starch, cellobiose, glucose, galactose, fructose and pyruvate, producing H2, acetate and CO2. Growth was observed from 50 to 80 degrees C with an optimum at 70 degrees C. The optimum pH was 6.8-7.1. The optimum concentration of sea salts in the medium was 20.5-25.5 g l(-1). The generation time under optimal conditions was 7.1 h. The DNA G+C content was 33 mol %. Growth of isolate JMT was not inhibited by penicillin, but ampicillin, streptomycin, kanamycin and neomycin completely inhibited growth. The results of 16S rDNA sequence analysis revealed that strain JMT belongs to the Thermoanaerobacter phylogenetic group within the Bacillus-Clostridium subphylum of Gram-positive bacteria but represents a separate branch of this group. On the basis of morphological and physiological features and phylogenetic data, this isolate should be assigned to a new genus, for which the name Carboxydobrachium is proposed. The type species is Carboxydobrachium pacificum; the type strain is JMT (= DSM 12653T).

  4. Dry-thermophilic anaerobic digestion of organic fraction of municipal solid waste: Methane production modeling

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Fdez-Gueelfo, L.A., E-mail: alberto.fdezguelfo@uca.es; Alvarez-Gallego, C.; Sales, D.

    2012-03-15

    Highlights: Black-Right-Pointing-Pointer Methane generation may be modeled by means of modified product generation model of Romero Garcia (1991). Black-Right-Pointing-Pointer Organic matter content and particle size influence the kinetic parameters. Black-Right-Pointing-Pointer Higher organic matter content and lower particle size enhance the biomethanization. - Abstract: The influence of particle size and organic matter content of organic fraction of municipal solid waste (OFMSW) in the overall kinetics of dry (30% total solids) thermophilic (55 Degree-Sign C) anaerobic digestion have been studied in a semi-continuous stirred tank reactor (SSTR). Two types of wastes were used: synthetic OFMSW (average particle size of 1 mm; 0.71more » g Volatile Solids/g waste), and OFMSW coming from a composting full scale plant (average particle size of 30 mm; 0.16 g Volatile Solids/g waste). A modification of a widely-validated product-generation kinetic model has been proposed. Results obtained from the modified-model parameterization at steady-state (that include new kinetic parameters as K, Y{sub pMAX} and {theta}{sub MIN}) indicate that the features of the feedstock strongly influence the kinetics of the process. The overall specific growth rate of microorganisms ({mu}{sub max}) with synthetic OFMSW is 43% higher compared to OFMSW coming from a composting full scale plant: 0.238 d{sup -1} (K = 1.391 d{sup -1}; Y{sub pMAX} = 1.167 L CH{sub 4}/gDOC{sub c}; {theta}{sub MIN} = 7.924 days) vs. 0.135 d{sup -1} (K = 1.282 d{sup -1}; Y{sub pMAX} = 1.150 L CH{sub 4}/gDOC{sub c}; {theta}{sub MIN} = 9.997 days) respectively. Finally, it could be emphasized that the validation of proposed modified-model has been performed successfully by means of the simulation of non-steady state data for the different SRTs tested with each waste.« less

  5. Garciella nitratireducens gen. nov., sp. nov., an anaerobic, thermophilic, nitrate- and thiosulfate-reducing bacterium isolated from an oilfield separator in the Gulf of Mexico.

    PubMed

    Miranda-Tello, Elizabeth; Fardeau, Marie-Laure; Sepúlveda, José; Fernández, Luis; Cayol, Jean-Luc; Thomas, Pierre; Ollivier, Bernard

    2003-09-01

    A novel Gram-positive, anaerobic and thermophilic bacterium, strain MET79(T), was isolated from an oil well located in the Gulf of Mexico. Cells were straight rods, motile by a subpolar flagellum. Spores were formed in old cultures. Inner gas vacuoles swelled the cells when exposed to air. The optimum growth conditions were 55 degrees C, pH 7.5 and 1 % NaCl. Yeast extract was required for growth. Strain MET79(T) fermented several sugars, some organic acids and Casamino acids. Glucose was fermented into lactate, acetate, butyrate, H(2) and CO(2). Strain MET79(T) reduced thiosulfate to hydrogen sulfide and nitrate to ammonium. The DNA G+C content was 30.9 mol%. The closest phylogenetic relative of strain MET79(T) was Caloranaerobacter azorensis (88.7 % 16S rDNA sequence similarity). As strain MET79(T) (=DSM 15102(T)=CIP 107615(T)) was physiologically and phylogenetically different from its closest relatives, it is assigned as the type strain of a novel species of a new genus, Garciella nitratireducens gen. nov., sp. nov.

  6. Anaerobic co-digestion of sewage sludge and food waste using temperature-phased anaerobic digestion process.

    PubMed

    Kim, H W; Han, S K; Shin, H S

    2004-01-01

    This study was performed to overcome the low efficiency of anaerobic digestion of sewage sludge and food waste by combining temperature-phased digestion, sequencing batch operation, and co-digestion technology. It was demonstrated that the temperature-phased anaerobic sequencing batch reactor (TPASBR) system for the co-digestion of sewage sludge and food waste resulted in enhanced volatile solids (VS) reduction and methane production rate. At the organic loading rate (OLR) of 2.7 g VS/l/d, the TPASBR system showed the higher VS reduction (61.3%), CH4 yield (0.28 l/g VS(added)) and CH4 production rate (0.41 l CH4/l/d) than those (0.29 l CH4/l/d) of the mesophilic two-stage ASBR (MTSASBR). In the specific methanogenic activity (SMA) tests on thermophilic biomass of the TPASBR system, the average SMA of acetate (93 ml CH4/gVSS/d) was much higher than those of propionate (46 ml CH4/g VSS/d) and butyrate (76 ml CH4/g VSS/d). Also, higher specific hydrolytic activity (SHA, 217 mg COD/g VSS/d) of the biomass supported fast hydrolysis under thermophilic conditions. The track study revealed that the most active period of the 24 h cycle was between 6 and 12 h. The enhanced performance of the TPASBR system could be attributed to longer solids retention time, fast hydrolysis, higher CH4 conversion rate, and balanced nutrient condition of co-substrate. It was verified that this combination could be a promising and practical alternative for the simultaneous recycling of two types of organic fraction of municipal solid waste (OFMSW) with high stability.

  7. Thermophilic biogasification of biomass

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Ghosh, S.; Klass, D.L.; Edwards, V.H.

    1980-01-01

    Secondary sewage effluent- and fresh-water-grown water hyacinths (Eichhornia crassipes), Coastal Bermuda grass (Cynodon dactylon), and a hyacinth-grass-municipal solid waste-sludge (biomass-waste) blend were used as test feeds to develop a fast thermophilic biomass- digestion process. For the pure biomass feeds thermophilic digestion has no apparent advantage over mesophilic digestion, but the reverse is true for the biomass-waste blend. Alkaline pretreatment of the feed improved thermophilic digester performance substantially. For a given plant feed load, the reactor volume, culture-heating requirements, and CH4 production rate for thermophilic digestion of the pretreated biomass-waste feed were 18,46, and 135% of those for conventional mesophilic digestion.more » For a biomass-waste feed the respective volatile solids reduction and energy recovery efficiencies were 46 and 49% for thermophilic and 36 and 43% for mesophilic digestions.« less

  8. Fervidicella metallireducens gen. nov., sp. nov., a thermophilic, anaerobic bacterium from geothermal waters.

    PubMed

    Ogg, Christopher D; Patel, Bharat K C

    2010-06-01

    A strictly anaerobic, thermophilic bacterium, designated strain AeB(T), was isolated from microbial mats colonizing a run-off channel formed by free-flowing thermal water from a bore well (registered number 17263) of the Great Artesian Basin, Australia. Cells of strain AeB(T) were slightly curved rods (2.5-6.0x1.0 mum) that stained Gram-negative and formed spherical terminal to subterminal spores. The strain grew optimally in tryptone-yeast extract-Casamino acids medium at 50 degrees C (range 37-55 degrees C) and pH 7 (range pH 5-9). Strain AeB(T) grew poorly on yeast extract (0.2 %) and tryptone (0.2 %) as sole carbon sources, which were obligately required for growth on other energy sources. Growth of strain AeB(T) increased in the presence of various carbohydrates and amino acids, but not organic acids. End products detected from glucose fermentation were ethanol, acetate, CO2 and H2. In the presence of 0.2 % yeast extract, iron(III), manganese(IV), vanadium(V) and cobalt(III) were reduced, but not sulfate, thiosulfate, sulfite, elemental sulfur, nitrate or nitrite. Iron(III) was also reduced in the presence of tryptone, peptone, Casamino acids and amyl media (Research Achievement), but not starch, xylan, chitin, glycerol, ethanol, pyruvate, benzoate, lactate, acetate, propionate, succinate, glycine, serine, lysine, threonine, arginine, glutamate, valine, leucine, histidine, alanine, aspartate, isoleucine or methionine. Growth was inhibited by chloramphenicol, streptomycin, tetracycline, penicillin, ampicillin and NaCl concentrations >2 %. The DNA G+C content was 35.4+/-1 mol%, as determined by the thermal denaturation method. 16S rRNA gene sequence analysis indicated that strain AeB(T) is a member of the family Clostridiaceae, class Clostridia, phylum 'Firmicutes', and is positioned approximately equidistantly between the genera Sarcina, Anaerobacter, Caloramator and Clostridium (16S rRNA gene similarity values of 87.8-90.9 %). On the basis of 16S rRNA gene

  9. Cellulose- and xylan-degrading thermophilic anaerobic bacteria from biocompost.

    PubMed

    Sizova, M V; Izquierdo, J A; Panikov, N S; Lynd, L R

    2011-04-01

    Nine thermophilic cellulolytic clostridial isolates and four other noncellulolytic bacterial isolates were isolated from self-heated biocompost via preliminary enrichment culture on microcrystalline cellulose. All cellulolytic isolates grew vigorously on cellulose, with the formation of either ethanol and acetate or acetate and formate as principal fermentation products as well as lactate and glycerol as minor products. In addition, two out of nine cellulolytic strains were able to utilize xylan and pretreated wood with roughly the same efficiency as for cellulose. The major products of xylan fermentation were acetate and formate, with minor contributions of lactate and ethanol. Phylogenetic analyses of 16S rRNA and glycosyl hydrolase family 48 (GH48) gene sequences revealed that two xylan-utilizing isolates were related to a Clostridium clariflavum strain and represent a distinct novel branch within the GH48 family. Both isolates possessed high cellulase and xylanase activity induced independently by either cellulose or xylan. Enzymatic activity decayed after growth cessation, with more-rapid disappearance of cellulase activity than of xylanase activity. A mixture of xylan and cellulose was utilized simultaneously, with a significant synergistic effect observed as a reduction of lag phase in cellulose degradation.

  10. Thermophilic lignocellulose deconstruction.

    PubMed

    Blumer-Schuette, Sara E; Brown, Steven D; Sander, Kyle B; Bayer, Edward A; Kataeva, Irina; Zurawski, Jeffrey V; Conway, Jonathan M; Adams, Michael W W; Kelly, Robert M

    2014-05-01

    Thermophilic microorganisms are attractive candidates for conversion of lignocellulose to biofuels because they produce robust, effective, carbohydrate-degrading enzymes and survive under harsh bioprocessing conditions that reflect their natural biotopes. However, no naturally occurring thermophile is known that can convert plant biomass into a liquid biofuel at rates, yields and titers that meet current bioprocessing and economic targets. Meeting those targets requires either metabolically engineering solventogenic thermophiles with additional biomass-deconstruction enzymes or engineering plant biomass degraders to produce a liquid biofuel. Thermostable enzymes from microorganisms isolated from diverse environments can serve as genetic reservoirs for both efforts. Because of the sheer number of enzymes that are required to hydrolyze plant biomass to fermentable oligosaccharides, the latter strategy appears to be the preferred route and thus has received the most attention to date. Thermophilic plant biomass degraders fall into one of two categories: cellulosomal (i.e. multienzyme complexes) and noncellulosomal (i.e. 'free' enzyme systems). Plant-biomass-deconstructing thermophilic bacteria from the genera Clostridium (cellulosomal) and Caldicellulosiruptor (noncellulosomal), which have potential as metabolic engineering platforms for producing biofuels, are compared and contrasted from a systems biology perspective. © 2013 Federation of European Microbiological Societies. Published by John Wiley & Sons Ltd. All rights reserved.

  11. Bacterial community analysis of swine manure treated with autothermal thermophilic aerobic digestion.

    PubMed

    Han, Il; Congeevaram, Shankar; Ki, Dong-Won; Oh, Byoung-Taek; Park, Joonhong

    2011-02-01

    Due to the environmental problems associated with disposal of livestock sludge, many stabilization studies emphasizing on the sludge volume reduction were performed. However, little is known about the microbial risk present in sludge and its stabilized products. This study microbiologically explored the effects of anaerobic lagoon fermentation (ALF) and autothermal thermophilic aerobic digestion (ATAD) on pathogen-related risk of raw swine manure by using culture-independent 16S rDNA cloning and sequencing methods. In raw swine manure, clones closely related to pathogens such as Dialister pneumosintes, Erysipelothrix rhusiopathiae, Succinivibrioan dextrinosolvens, and Schineria sp. were detected. Meanwhile, in the mesophilic ALF-treated swine manure, bacterial community clones closely related to pathogens such as Schineria sp. and Succinivibrio dextrinosolvens were still detected. Interestingly, the ATAD treatment resulted in no detection of clones closely related to pathogens in the stabilized thermophilic bacterial community, with the predominance of novel Clostridia class populations. These findings support the superiority of ATAD in selectively reducing potential human and animal pathogens compared to ALF, which is a typical manure stabilization method used in livestock farms.

  12. Themoanaerobacterium calidifontis sp. nov., a novel anaerobic, thermophilic, ethanol-producing bacterium from hot springs in China.

    PubMed

    Shang, Shu-mei; Qian, Long; Zhang, Xu; Li, Kun-zhi; Chagan, Irbis

    2013-06-01

    A novel thermophilic Gram staining positive strain Rx1 was isolated from hot springs in Baoshan of Yunnan Province, China. The strain was characterized as a hemicellulose-decomposing obligate anaerobe bacterium that is rod-shaped (diameter: 0.5-0.7 μm; length: 2.0-6.7 μm), spore-forming, and motile. Its growth temperature range is 38-68 °C (optimum 50-55 °C) and pH range is 4.5-8.0 (optimum 7.0). The maximum tolerance concentration of NaCl was 3 %. Rx1 converted thiosulfate to elemental sulfur and reduced sulfite to hydrogen sulfide. The bacterium grew by utilizing xylan and starch, as well as a wide range of monosaccharide and polysaccharides, including glucose and xylose. The main products of fermentation were ethanol, lactate, acetate, CO2, and H2. The maximum xylanase activity in the culture supernatant after 30 h of incubation at 55 °C was 16.2 U/ml. Rx1 DNA G + C content was 36 mol %. 16S rRNA gene sequence analysis indicated that strain Rx1 belonged to the genus Thermoanaerobacterium of the family 'Thermoanaerobacteriaceae' (Firmicutes), with Thermoanaerobacterium aciditolerans 761-119 (99.2 % 16S rRNA gene sequence similarity) being its closest relative. DNA-DNA hybridization between Rx1 and T. aciditolerans 761-119 showed 36 % relatedness. Based on its physiological and biochemical tests and DNA-DNA hybridization analyses, the isolate is considered to represent a novel species in the genus Thermoanaerobacterium, for which the name Thermoanaerobacterium calidifontis sp. nov. is proposed, with the type strain is Rx1 (=JCM 18270 = CCTCC M 2011109).

  13. Carboxydocella sporoproducens sp. nov., a novel anaerobic CO-utilizing/H2-producing thermophilic bacterium from a Kamchatka hot spring.

    PubMed

    Slepova, Tatiana V; Sokolova, Tatyana G; Lysenko, Anatoly M; Tourova, Tatyana P; Kolganova, Tatyana V; Kamzolkina, Olga V; Karpov, Genady A; Bonch-Osmolovskaya, Elizaveta A

    2006-04-01

    A novel anaerobic, thermophilic, CO-utilizing bacterium, strain KarT, was isolated from a hot spring of Karymskoe Lake, Kamchatka Peninsula. The cells of the novel isolate were Gram-positive, spore-forming, short rods. The bacterium grew chemolithoautotrophically on CO, producing equimolar quantities of H2 and CO2 (according to the equation CO + H2O --> CO2 + H2), and in the absence of CO, under N2 in the gas phase, chemoorganoheterotrophically with yeast extract, sucrose or pyruvate. Growth was observed in the temperature range 50-70 degrees C, with an optimum at 60 degrees C, and in the pH range 6.2-8.0, with an optimum at pH 6.8. The micro-organism did not grow on solid media; it was able to grow only in semi-solid medium containing 0.5 % agar. The generation time under optimal conditions for chemolithoautotrophic growth was 1 h. The G+C content of the DNA was 46.5+/-1 mol%. Growth was completely inhibited by penicillin, novobiocin, streptomycin, kanamycin and neomycin. Analysis of the 16S rRNA gene sequence showed that the isolate should be assigned to the genus Carboxydocella. On the basis of the results of DNA-DNA hybridization and morphological and physiological analyses, strain KarT represents a novel species of the genus Carboxydocella, for which the name Carboxydocella sporoproducens sp. nov. is proposed. The type strain is KarT (=DSM 16521T = VKM B-2358T).

  14. Thermostable lipases from the extreme thermophilic anaerobic bacteria Thermoanaerobacter thermohydrosulfuricus SOL1 and Caldanaerobacter subterraneus subsp. tengcongensis.

    PubMed

    Royter, Marina; Schmidt, M; Elend, C; Höbenreich, H; Schäfer, T; Bornscheuer, U T; Antranikian, G

    2009-09-01

    Two novel genes encoding for heat and solvent stable lipases from strictly anaerobic extreme thermophilic bacteria Thermoanaerobacter thermohydrosulfuricus (LipTth) and Caldanaerobacter subterraneus subsp. tengcongensis (LipCst) were successfully cloned and expressed in E. coli. Recombinant proteins were purified to homogeneity by heat precipitation, hydrophobic interaction, and gel filtration chromatography. Unlike the enzymes from mesophile counterparts, enzymatic activity was measured at a broad temperature and pH range, between 40 and 90 degrees C and between pH 6.5 and 10; the half-life of the enzymes at 75 degrees C and pH 8.0 was 48 h. Inhibition was observed with 4-(2-aminoethyl)-benzenesulfonyl fluoride hydrochloride and phenylmethylsulfonylfluorid indicating that serine and thiol groups play a role in the active site of the enzymes. Gene sequence comparisons indicated very low identity to already described lipases from mesophilic and psychrophilic microorganisms. By optimal cultivation of E. coli Tuner (DE3) cells in 2-l bioreactors, a massive production of the recombinant lipases was achieved (53-2200 U/l) Unlike known lipases, the purified robust proteins are resistant against a large number of organic solvents (up to 99%) and detergents, and show activity toward a broad range of substrates, including triacylglycerols, monoacylglycerols, esters of secondary alcohols, and p-nitrophenyl esters. Furthermore, the enzyme from T. thermohydrosulfuricus is suitable for the production of optically pure compounds since it is highly S-stereoselective toward esters of secondary alcohols. The observed E values for but-3-yn-2-ol butyrate and but-3-yn-2-ol acetate of 21 and 16, respectively, make these enzymes ideal candidates for kinetic resolution of synthetically useful compounds.

  15. Thermophilic biogasification of biomass

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Ghosh, S.; Klass, D.L.; Christopher, R.W.

    1980-01-01

    Mesophilic and thermophilic digestion runs were conducted with the pure land-based biomass species, water hyacinth (Eichhornia crassipes) and Coastal Bermuda grass (Cynodon dactylon), and a blend of hyacinth, grass, MSW, and sewage sludge. A mixed biomass-waste hybrid feed was included because it has a superior nutritional balance relative to the pure feeds and it facilitates year-round operation of a biomass-to-SNG process. (7) The studies were conducted at 35/sup 0/ and 55/sup 0/C, generally believed to be optimum for mesophilic and thermophilic digestion of organic feeds. Results of mesophilic digestion were to provide baseline performance data for evaluation of thermophilic digestermore » performance. It was decided that the feed affording the best thermophilic performance would be pretreated with dilute sodium hydroxide solution at the selected digestion temperature of 55/sup 0/C to improve methane production rate and yield. In addition, thermophilic runs were planned to investigate ways to reduce chemical requirements for alkaline pretreatment and feed slurry neutralization.« less

  16. Anaerobic digestion of sulfate-acidified cattle slurry: One-stage vs. two-stage.

    PubMed

    Moset, Veronica; Ottosen, Lars Ditlev Mørck; Xavier, Cristiane de Almeida Neves; Møller, Henrik Bjarne

    2016-05-15

    Two strategies to include acidified cattle manure (AcCM) in co-digestion with normal cattle manure (CM) are presented in this work. The strategies are a single thermophilic (50 °C) continuous stirred tank reactor (CSTR) anaerobic digestion and a two-step (65 °C + 50 °C) CSTR process. In both strategies, two different inclusion levels of H2SO4-acidified CM (10% and 20%) in co-digestion with normal CM were tested and compared with a control CSTR fed only CM. Important enhancement of methane (CH4) yield and solid reductions were observed in the thermophilic one-step CSTR working with 10% AcCM. However, a higher inclusion level of AcCM (20%) caused volatile fatty acid accumulation in the reactor and a more than 30% reduction in CH4 production. In terms of CH4 production, when 10% of AcCM was co-digested with 90% of CM, the two-step anaerobic co-digestion yielded less than the single step. During the first step of the two-step CSTR process, acidogenesis and a partial sulfate reduction were achieved. However, sulfide stripping between the first and the second step must be promoted in order to advance this technology. Copyright © 2016 Elsevier Ltd. All rights reserved.

  17. Anaerobic bioprocessing of organic wastes.

    PubMed

    Verstraete, W; de Beer, D; Pena, M; Lettinga, G; Lens, P

    1996-05-01

    continuously stirred tank system to the thermophilic configuration, as the latter permits higher conversion rates and easier sanitation. Integration of ultrafiltration in anaerobic slurry digestion facilitates operation at higher volumetric loading rates and at shorter residence times. With respect to organic solids, the recent trend in society towards source separated collection of biowaste has opened a broad range of new application areas for solid state anaerobic fermentation.

  18. Enhanced Versus Conventional Sludge Anaerobic Processes: Performances and Techno-Economic Assessment.

    PubMed

    Gianico, Andrea; Bertanza, Giorgio; Braguglia, Camilla M; Canato, Matteo; Gallipoli, Agata; Laera, Giuseppe; Levantesi, Caterina; Mininni, Giuseppe

    2016-05-01

    Sewage sludge processing is a key issue in water resource recovery facilities due to the inefficacy of conventional treatments to produce high quality biosolids to be safely used in agriculture. Under this framework, the performances of several enhanced stabilization processes, namely ultrasound-pretreated Mesophilic Anaerobic Digestion (US+MAD), thermophilic anaerobic digestion (TAD), thermal-pretreated TAD (TH+TAD) and ultrasound-pretreated inverse Temperature Phased Anaerobic Digestion (US+iTPAD) have been investigated. Such enhanced processes resulted in higher biogas yields and higher destruction of pathogens with respect to conventional MAD process, thus suggesting their feasibility in full-scale implementation perspectives. A procedure for technical-economic comparison of new sludge processing lines against conventional ones (benchmarking) was developed, based on the definition of technical issues (e.g. reliability, complexity, etc.) which are rated for each situation. Moreover, capital and operating costs were estimated. The enhanced processes analyzed in this work showed some potentially critical items, mainly related to energy balance and reagent consumption.

  19. Thermophilic molds: Biology and applications.

    PubMed

    Singh, Bijender; Poças-Fonseca, Marcio J; Johri, B N; Satyanarayana, Tulasi

    2016-11-01

    Thermophilic molds thrive in a variety of natural habitats including soils, composts, wood chip piles, nesting materials of birds and other animals, municipal refuse and others, and ubiquitous in their distribution. These molds grow in simple media containing carbon and nitrogen sources and mineral salts. Polyamines are synthesized in these molds and the composition of lipids varies considerably, predominantly containing palmitic, oleic and linoleic acids with low levels of lauric, palmiotoleic and stearic acids. Thermophilic molds are capable of efficiently degrading organic materials by secreting thermostable enzymes, which are useful in the bioremediation of industrial wastes and effluents that are rich in oil, heavy metals, anti-nutritional factors such as phytic acid and polysaccharides. Thermophilic molds synthesize several antimicrobial substances and biotechnologically useful miscellaneous enzymes. The analysis of genomes of thermophilic molds reveals high G:C contents, shorter introns and intergenic regions with lesser repetitive sequences, and further confirms their ability to degrade agro-residues efficiently. Genetic engineering has aided in ameliorating the characteristics of the enzymes of thermophilic molds. This review is aimed at focusing on the biology of thermophilic molds with emphasis on recent developments in the analysis of genomes, genetic engineering and potential applications.

  20. Combined mesophilic anaerobic and thermophilic aerobic digestion process for high-strength food wastewater to increase removal efficiency and reduce sludge discharge.

    PubMed

    Jang, H M; Park, S K; Ha, J H; Park, J M

    2014-01-01

    In this study, a process that combines the mesophilic anaerobic digestion (MAD) process with thermophilic aerobic digestion (TAD) for high-strength food wastewater (FWW) treatment was developed to examine the removal of organic matter and methane production. All effluent discharged from the MAD process was separated into solid and liquid portions. The liquid part was discarded and the sludge part was passed to the TAD process for further degradation. Then, the digested sludge from the TAD process was recycled back to the MAD unit to achieve low sludge discharge from the combined process. The reactor combination was operated in two phases: during Phase I, 40 d of total hydraulic retention time (HRT) was applied; during Phase II, 20 d was applied. HRT of the TAD process was fixed at 5 d. For a comparison, a control process (single-stage MAD) was operated with the same HRTs of the combined process. Our results indicated that the combined process showed over 90% total solids, volatile solids and chemical oxygen demand removal efficiencies. In addition, the combined process showed a significantly higher methane production rate than that of the control process. Consequently, the experimental data demonstrated that the combined MAD-TAD process was successfully employed for high-strength FWW treatment with highly efficient organic matter reduction and methane production.

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

    NASA Astrophysics Data System (ADS)

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

    2016-12-01

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

  2. Thermophilic, anaerobic co-digestion of microalgal biomass and cellulose for H2 production.

    PubMed

    Carver, Sarah M; Hulatt, Chris J; Thomas, David N; Tuovinen, Olli H

    2011-07-01

    Microalgal biomass has been a focus in the sustainable energy field, especially biodiesel production. The purpose of this study was to assess the feasibility of treating microalgal biomass and cellulose by anaerobic digestion for H2 production. A microbial consortium, TC60, known to degrade cellulose and other plant polymers, was enriched on a mixture of cellulose and green microalgal biomass of Dunaliella tertiolecta, a marine species, or Chlorella vulgaris, a freshwater species. After five enrichment steps at 60°C, hydrogen yields increased at least 10% under all conditions. Anaerobic digestion of D. tertiolecta and cellulose by TC60 produced 7.7 mmol H2/g volatile solids (VS) which were higher than the levels (2.9-4.2 mmol/g VS) obtained with cellulose and C. vulgaris biomass. Both microalgal slurries contained satellite prokaryotes. The C. vulgaris slurry, without TC60 inoculation, generated H2 levels on par with that of TC60 on cellulose alone. The biomass-fed anaerobic digestion resulted in large shifts in short chain fatty acid concentrations and increased ammonium levels. Growth and H2 production increased when TC60 was grown on a combination of D. tertiolecta and cellulose due to nutrients released from algal cells via lysis. The results indicated that satellite heterotrophs from C. vulgaris produced H2 but the Chlorella biomass was not substantially degraded by TC60. To date, this is the first study to examine H2 production by anaerobic digestion of microalgal biomass. The results indicate that H2 production is feasible but higher yields could be achieved by optimization of the bioprocess conditions including biomass pretreatment.

  3. Nautilia nitratireducens sp. nov., a thermophilic, anaerobic, chemosynthetic, nitrate-ammonifying bacterium isolated from a deep-sea hydrothermal vent.

    PubMed

    Pérez-Rodríguez, Ileana; Ricci, Jessica; Voordeckers, James W; Starovoytov, Valentin; Vetriani, Costantino

    2010-05-01

    A thermophilic, anaerobic, chemosynthetic bacterium, designated strain MB-1(T), was isolated from the walls of an active deep-sea hydrothermal vent chimney on the East Pacific Rise at degrees 50' N 10 degrees 17' W. The cells were Gram-negative-staining rods, approximately 1-1.5 mum long and 0.3-0.5 mum wide. Strain MB-1(T) grew at 25-65 degrees C (optimum 55 degrees C), with 10-35 g NaCl l(-1) (optimum 20 g l(-1)) and at pH 4.5-8.5 (optimum pH 7.0). Generation time under optimal conditions was 45.6 min. Growth occurred under chemolithoautotrophic conditions with H(2) as the energy source and CO(2) as the carbon source. Nitrate was used as the electron acceptor, with resulting production of ammonium. Thiosulfate, sulfur and selenate were also used as electron acceptors. No growth was observed in the presence of lactate, peptone or tryptone. Chemo-organotrophic growth occurred in the presence of acetate, formate, Casamino acids, sucrose, galactose and yeast extract under a N(2)/CO(2) gas phase. The G+C content of the genomic DNA was 36.0 mol%. Phylogenetic analysis of the 16S rRNA gene sequence indicated that this organism is closely related to Nautilia profundicola AmH(T), Nautilia abyssi PH1209(T) and Nautilia lithotrophica 525(T) (95, 94 and 93 % sequence identity, respectively). On the basis of phylogenetic, physiological and genetic considerations, it is proposed that the organism represents a novel species within the genus Nautilia, Nautilia nitratireducens sp. nov. The type strain is MB-1(T) (=DSM 22087(T) =JCM 15746(T)).

  4. Hydrogen Limitation, Competition and Syntrophy Among Thermophiles in the Hot Subseafloor

    NASA Astrophysics Data System (ADS)

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

    2016-02-01

    To understand the biogeochemical impact of subseafloor life on surrounding environments and the overlying ocean, the rates of and constraints on growth and metabolite production must be determined for subseafloor microbes. The diversity of thermophiles and hyperthermophiles in the hot subseafloor is low, making them amenable to biogeochemical modeling. At hydrothermal vents, Methanothermococcus and Methanocaldococcus are common thermophilic and hyperthermophilic methanogens; Desulfurobacterium, common thermophilic autotrophic sulfur reducers; and Thermococcus, common hyperthermophilic H2-producing heterotrophs. Diffuse hydrothermal fluids (7-40°C) from Axial Seamount were incubated anaerobically at 55°C and 80°C. Microcosms with 1.2 mM added H2 produced 30-50 times more CH4 than those with 20 µM H2, demonstrating that H2 and heat were the primary constraints on the growth of thermophilic and hyperthermophilic methanogens. For Methanothermococcus thermolithotrophicus and Methanocaldococcus jannaschii, the lower H2 thresholds and half-saturation constants (Ks) for growth in a chemostat were 9-20 µM and 47-69 µM, respectively, near their optimum growth temperatures. The lower H2 threshold and Ks for Desulfurobacterium sp. strain HR11 grown in a batch reactor at 72°C were <3 µM and 29 µM, respectively. Its catalytic efficiency (µmax/Ks) for H2 use was 4 times higher than those of M. thermolithotrophicus and M. jannaschii, but its Ks for thiosulfate was 19 µM, suggesting that it will only outcompete methanogens when sulfur is not growth limiting. Microcosm incubations at 55°C and 80°C supplemented with peptides in lieu of H2 produced the same amount of CH4 as microcosms with 1.2 mM H2. Co-culture incubations showed that H2 production by T. paralvinellae supported the growth of Methanothermococcus sp. strain BW11, Methanocaldococcus bathoardescens, and strain HR11 in H2-free medium, suggesting that syntrophy is a possible alternative H2 source in the

  5. [Gradient elevation of temperature startup experiment of thermophilic ASBR treating thermal-hydrolyzed sewage sludge].

    PubMed

    Ouyang, Er-Ming; Wang, Wei; Long, Neng; Li, Huai

    2009-04-15

    Startup experiment was conducted for thermophilic anaerobic sequencing batch reactor (ASBR) treating thermal-hydrolyzed sewage sludge using the strategy of the step-wise temperature increment: 35 degrees C-->40 degrees C-->47 degrees C-->53 degrees C. The results showed that the first step-increase (from 35 degrees C to 40 degrees C) and final step-increase (from 47 degrees C to 53 degrees C) had only a slight effect on the digestion process. The second step-increase (from 40 degrees C to 47 degrees C) resulted in a severe disturbance: the biogas production, methane content, CODeffluent and microorganism all have strong disturbance. At the steady stage of thermophilic ASBR treating thermal-hydrolyzed sewage sludge, the average daily gas production, methane content, specific methane production (CH4/CODinfluent), TCOD removal rate and SCOD removal rate were 2.038 L/d, 72.0%, 188.8 mL/g, 63.8%, 83.3% respectively. The results of SEM and DGGE indicated that the dominant species are obviously different at early stage and steady stage.

  6. Early Microbial Evolution: The Age of Anaerobes

    PubMed Central

    Martin, William F.; Sousa, Filipa L.

    2016-01-01

    In this article, the term “early microbial evolution” refers to the phase of biological history from the emergence of life to the diversification of the first microbial lineages. In the modern era (since we knew about archaea), three debates have emerged on the subject that deserve discussion: (1) thermophilic origins versus mesophilic origins, (2) autotrophic origins versus heterotrophic origins, and (3) how do eukaryotes figure into early evolution. Here, we revisit those debates from the standpoint of newer data. We also consider the perhaps more pressing issue that molecular phylogenies need to recover anaerobic lineages at the base of prokaryotic trees, because O2 is a product of biological evolution; hence, the first microbes had to be anaerobes. If molecular phylogenies do not recover anaerobes basal, something is wrong. Among the anaerobes, hydrogen-dependent autotrophs—acetogens and methanogens—look like good candidates for the ancestral state of physiology in the bacteria and archaea, respectively. New trees tend to indicate that eukaryote cytosolic ribosomes branch within their archaeal homologs, not as sisters to them and, furthermore tend to root archaea within the methanogens. These are major changes in the tree of life, and open up new avenues of thought. Geochemical methane synthesis occurs as a spontaneous, abiotic exergonic reaction at hydrothermal vents. The overall similarity between that reaction and biological methanogenesis fits well with the concept of a methanogenic root for archaea and an autotrophic origin of microbial physiology. PMID:26684184

  7. Winery waste recycling through anaerobic co-digestion with waste activated sludge.

    PubMed

    Da Ros, C; Cavinato, C; Pavan, P; Bolzonella, D

    2014-11-01

    In this study biogas and high quality digestate were recovered from winery waste (wine lees) through anaerobic co-digestion with waste activated sludge both in mesophilic and thermophilic conditions. The two conditions studied showed similar yields (0.40 m(3)/kgCODfed) but different biological process stability: in fact the mesophilic process was clearly more stable than the thermophilic one in terms of bioprocess parameters. The resulting digestates showed good characteristics for both the tested conditions: heavy metals, dioxins (PCDD/F), and dioxin like bi-phenyls (PCBs) were concentred in the effluent if compared with the influent because of the important reduction of the solid dry matter, but remained at levels acceptable for agricultural reuse. Pathogens in digestate decreased. Best reductions were observed in thermophilic condition, while at 37°C the concentration of Escherichia coli was at concentrations level as high as 1000 UFC/g. Dewatering properties of digestates were evaluated by means of the capillary suction time (CST) and specific resistance to filtration (SRF) tests and it was found that a good dewatering level was achievable only when high doses of polymer (more than 25 g per kg dry solids) were added to sludge. Copyright © 2014 Elsevier Ltd. All rights reserved.

  8. Enhanced mesophilic anaerobic digestion of food waste by thermal pretreatment: Substrate versus digestate heating.

    PubMed

    Ariunbaatar, Javkhlan; Panico, Antonio; Yeh, Daniel H; Pirozzi, Francesco; Lens, Piet N L; Esposito, Giovanni

    2015-12-01

    Food waste (FW) represents a source of high potential renewable energy if properly treated with anaerobic digestion (AD). Pretreating the substrates could yield a higher biomethane production in a shorter time. In this study, the effects of thermal (heating the FW in a separate chamber) and thermophilic (heating the full reactor content containing both FW and inoculum) pretreatments at 50, 60, 70 and 80°C prior to mesophilic AD were studied through a series of batch experiments. Pretreatments at a lower temperature (50°C) and a shorter time (<12h) had a positive effect on the AD process. The highest enhancement of the biomethane production with an increase by 44-46% was achieved with a thermophilic pretreatment at 50°C for 6-12h or a thermal pretreatment at 80°C for 1.5h. Thermophilic pretreatments at higher temperatures (>55°C) and longer operating times (>12h) yielded higher soluble chemical oxygen demand (CODs), but had a negative effect on the methanogenic activity. The thermal pretreatments at the same conditions resulted in a lower solubilization of COD. Based on net energy calculations, the enhanced biomethane production is sufficient to heat up the FW for the thermal, but not for the thermophilic pretreatment. Copyright © 2015 Elsevier Ltd. All rights reserved.

  9. Boosting dark fermentation with co-cultures of extreme thermophiles for biohythane production from garden waste.

    PubMed

    Abreu, Angela A; Tavares, Fábio; Alves, Maria Madalena; Pereira, Maria Alcina

    2016-11-01

    Proof of principle of biohythane and potential energy production from garden waste (GW) is demonstrated in this study in a two-step process coupling dark fermentation and anaerobic digestion. The synergistic effect of using co-cultures of extreme thermophiles to intensify biohydrogen dark fermentation is demonstrated using xylose, cellobiose and GW. Co-culture of Caldicellulosiruptor saccharolyticus and Thermotoga maritima showed higher hydrogen production yields from xylose (2.7±0.1molmol(-1) total sugar) and cellobiose (4.8±0.3molmol(-1) total sugar) compared to individual cultures. Co-culture of extreme thermophiles C. saccharolyticus and Caldicellulosiruptor bescii increased synergistically the hydrogen production yield from GW (98.3±6.9Lkg(-1) (VS)) compared to individual cultures and co-culture of T. maritima and C. saccharolyticus. The biochemical methane potential of the fermentation end-products was 322±10Lkg(-1) (CODt). Biohythane, a biogas enriched with 15% hydrogen could be obtained from GW, yielding a potential energy generation of 22.2MJkg(-1) (VS). Copyright © 2016 Elsevier Ltd. All rights reserved.

  10. Thermotalea metallivorans gen. nov., sp. nov., a thermophilic, anaerobic bacterium from the Great Artesian Basin of Australia aquifer.

    PubMed

    Ogg, Christopher D; Patel, Bharat K C

    2009-05-01

    A strictly anaerobic, thermophilic bacterium, designated strain B2-1(T), was isolated from microbial mats colonizing a runoff channel formed by free-flowing thermal water from a Great Artesian Basin, Australia, bore well (registered number 17263). The cells of strain B2-1(T) were slightly curved rods (3.0-3.5 x 0.6-0.7 microm) which stained Gram-negative. The strain grew optimally in tryptone-yeast extract-glucose medium at 50 degrees C (temperature growth range 30-55 degrees C) and a pH of 8 (pH growth range 6.5-9). Strain B2-1(T) grew poorly on yeast extract (0.2 %) and/or tryptone (0.2 %), which were obligately required for growth on other energy sources, including a range of other carbohydrates and organic acids, but not amino acids. The end-products of glucose fermentation were ethanol and acetate. In the presence of 0.2 % yeast extract, iron(III), manganese(IV) and elemental sulfur were reduced but sulfate, thiosulfate, sulfite, nitrate and nitrite were not reduced. Growth was inhibited by chloramphenicol, streptomycin, tetracycline, penicillin, ampicillin, sodium azide and by NaCl concentrations greater than 4 % (w/v). The DNA G+C content was 48+/-1 mol% as determined by the thermal denaturation method. 16S rRNA gene sequence analysis indicated that strain B2-1(T) was a member of the family Clostridiaceae, class Clostridia, phylum Firmicutes and was most closely related to Geosporobacter subterraneus DSM 17957(T) (89.9 % similarity). On the basis of 16S rRNA gene sequence comparisons and physiological characteristics, strain B2-1(T) is considered to represent a novel species of a new genus, for which the name Thermotalea metallivorans gen. nov., sp. nov. is proposed. The type strain is B2-1(T) (=KCTC 5625(T)=JCM 15105(T)=DSM 21119(T)).

  11. High organic loading rate on thermophilic hydrogen production and metagenomic study at an anaerobic packed-bed reactor treating a residual liquid stream of a Brazilian biorefinery.

    PubMed

    Ferraz Júnior, Antônio Djalma Nunes; Etchebehere, Claudia; Zaiat, Marcelo

    2015-06-01

    This study evaluated the influence of a high organic loading rate (OLR) on thermophilic hydrogen production at an up-flow anaerobic packed-bed reactor (APBR) treating a residual liquid stream of a Brazilian biorefinery. The APBR, filled with low-density polyethylene, was operated at an OLR of 84.2 kg-COD m(-3) d(-1). This value was determined in a previous study. The maximum values of hydrogen production and yield were 5,252.6 mL-H2 d(-1) and 3.7 mol-H2 mol(-1)(total carbohydrates), respectively. However, whereas the OLR remained constant, the specific organic load rate (sOLR) decreased throughout operation from 1.38 to 0.72 g-Total carbohydratesg-VS(-1) h(-1), this decrease negatively affected hydrogen production. A sOLR of 0.98 g-Total carbohydratesg-VS(-1) h(-1) was optimal for hydrogen production. The microbial community was studied using 454-pyrosequencing analysis. Organisms belonging to the genera Caloramator, Clostridium, Megasphaera, Oxobacter, Thermoanaerobacterium, and Thermohydrogenium were detected in samples taken from the reactor at operation days 30 and 60, suggesting that these organisms contribute to hydrogen production. Copyright © 2015 Elsevier Ltd. All rights reserved.

  12. Chemical Changes during Anaerobic Decomposition of Hardwood, Softwood, and Old Newsprint under Mesophilic and Thermophilic Conditions

    Treesearch

    Florentino B. De la Cruz; Daniel J. Yelle; Hanna S. Gracz; Morton A. Barlaz

    2014-01-01

    The anaerobic decomposition of plant biomass is an important aspect of global organic carbon cycling. While the anaerobic metabolism of cellulose and hemicelluloses to methane and carbon dioxide are well-understood, evidence for the initial stages of lignin decomposition is fragmentary. The objective of this study was to look for evidence of chemical transformations of...

  13. Thermophilic Fungi: Their Physiology and Enzymes†

    PubMed Central

    Maheshwari, Ramesh; Bharadwaj, Girish; Bhat, Mahalingeshwara K.

    2000-01-01

    Thermophilic fungi are a small assemblage in mycota that have a minimum temperature of growth at or above 20°C and a maximum temperature of growth extending up to 60 to 62°C. As the only representatives of eukaryotic organisms that can grow at temperatures above 45°C, the thermophilic fungi are valuable experimental systems for investigations of mechanisms that allow growth at moderately high temperature yet limit their growth beyond 60 to 62°C. Although widespread in terrestrial habitats, they have remained underexplored compared to thermophilic species of eubacteria and archaea. However, thermophilic fungi are potential sources of enzymes with scientific and commercial interests. This review, for the first time, compiles information on the physiology and enzymes of thermophilic fungi. Thermophilic fungi can be grown in minimal media with metabolic rates and growth yields comparable to those of mesophilic fungi. Studies of their growth kinetics, respiration, mixed-substrate utilization, nutrient uptake, and protein breakdown rate have provided some basic information not only on thermophilic fungi but also on filamentous fungi in general. Some species have the ability to grow at ambient temperatures if cultures are initiated with germinated spores or mycelial inoculum or if a nutritionally rich medium is used. Thermophilic fungi have a powerful ability to degrade polysaccharide constituents of biomass. The properties of their enzymes show differences not only among species but also among strains of the same species. Their extracellular enzymes display temperature optima for activity that are close to or above the optimum temperature for the growth of organism and, in general, are more heat stable than those of the mesophilic fungi. Some extracellular enzymes from thermophilic fungi are being produced commercially, and a few others have commercial prospects. Genes of thermophilic fungi encoding lipase, protease, xylanase, and cellulase have been cloned and

  14. Myceliophthora thermophila syn. Sporotrichum thermophile: a thermophilic mould of biotechnological potential.

    PubMed

    Singh, Bijender

    2016-01-01

    Myceliophthora thermophila syn. Sporotrichum thermophile is a ubiquitous thermophilic mould with a strong ability to degrade organic matter during optimal growth at 45 °C. Both genome analysis and experimental data have suggested that the mould is capable of hydrolyzing all major polysaccharides found in biomass. The mould is able to secrete a large number of hydrolytic enzymes (cellulases, laccases, xylanases, pectinases, lipases, phytases and some other miscellaneous enzymes) employed in various biotechnological applications. Characterization of the biomass-hydrolyzing activity of wild and recombinant enzymes suggests that this mould is highly efficient in biomass decomposition at both moderate and high temperatures. The native enzymes produced by the mould are more efficient in activity than their mesophilic counterparts beside their low enzyme titers. The mould is able to synthesize various biomolecules, which are used in multifarious applications. Genome sequence data of M. thermophila also supported the physiological data. This review describes the biotechnological potential of thermophilic mould, M. thermophila supported by genomic and experimental evidences.

  15. Digester performance and microbial community changes in thermophilic and mesophilic sequencing batch reactors fed with the fine sieved fraction of municipal sewage.

    PubMed

    Ghasimi, Dara S M; Tao, Yu; de Kreuk, Merle; Abbas, Ben; Zandvoort, Marcel H; van Lier, Jules B

    2015-12-15

    This study investigates the start-up and operation of bench-scale mesophilic (35 °C) and thermophilic (55 °C) anaerobic sequencing batch reactor (SBR) digesters treating the fine sieved fraction (FSF) from raw municipal sewage. FSF was sequestered from raw municipal wastewater, in the Netherlands, using a rotating belt filter equipped with a 350 micron mesh. For the given wastewater, the major component of FSF was toilet paper, which is estimated to be 10-14 kg per year per average person in the western European countries. A seven months adaptation time was allowed for the thermophilic and mesophilic digesters in order to adapt to FSF as the sole substrate with varying dry solids content of 10-25%. Different SBR cycle durations (14, 9 and 2 days) were applied for both temperature conditions to study methane production rates, volatile fatty acids (VFAs) dynamics, lag phases, as well as changes in microbial communities. The prevailing sludge in the two digesters consisted of very different bacterial and archaeal communities, with OP9 lineage and Methanothermobacter being pre-dominant in the thermophilic digester and Bacteroides and Methanosaeta dominating the mesophilic one. Eventually, decreasing the SBR cycle period, thus increasing the FSF load, resulted in improved digester performances, particularly with regard to the thermophilic digester, i.e. shortened lag phases following the batch feedings, and reduced VFA peaks. Over time, the thermophilic digester outperformed the mesophilic one with 15% increased volatile solids (VS) destruction, irrespective to lower species diversity found at high temperature. Copyright © 2015 Elsevier Ltd. All rights reserved.

  16. Thermophilic microorganisms in biomining.

    PubMed

    Donati, Edgardo Rubén; Castro, Camila; Urbieta, María Sofía

    2016-11-01

    Biomining is an applied biotechnology for mineral processing and metal extraction from ores and concentrates. This alternative technology for recovering metals involves the hydrometallurgical processes known as bioleaching and biooxidation where the metal is directly solubilized or released from the matrix for further solubilization, respectively. Several commercial applications of biomining can be found around the world to recover mainly copper and gold but also other metals; most of them are operating at temperatures below 40-50 °C using mesophilic and moderate thermophilic microorganisms. Although biomining offers an economically viable and cleaner option, its share of the world´s production of metals has not grown as much as it was expected, mainly considering that due to environmental restrictions in many countries smelting and roasting technologies are being eliminated. The slow rate of biomining processes is for sure the main reason of their poor implementation. In this scenario the use of thermophiles could be advantageous because higher operational temperature would increase the rate of the process and in addition it would eliminate the energy input for cooling the system (bioleaching reactions are exothermic causing a serious temperature increase in bioreactors and inside heaps that adversely affects most of the mesophilic microorganisms) and it would decrease the passivation of mineral surfaces. In the last few years many thermophilic bacteria and archaea have been isolated, characterized, and even used for extracting metals. This paper reviews the current status of biomining using thermophiles, describes the main characteristics of thermophilic biominers and discusses the future for this biotechnology.

  17. Biohydrogen and methane production by co-digestion of cassava stillage and excess sludge under thermophilic condition.

    PubMed

    Wang, Wen; Xie, Li; Chen, Jinrong; Luo, Gang; Zhou, Qi

    2011-02-01

    Thermophilic anaerobic hydrogen and methane production by co-digestion of cassava stillage (CS) and excess sludge (ES) was investigated in this study. The improved hydrogen and subsequent methane production were observed by co-digestion of CS with certain amount of ES in batch experiments. Compared with one phase anaerobic digestion, two phase anaerobic digestion offered an attractive alternative with more abundant biogas production and energy yield, e.g., the total energy yield in two phase obtained at VS(CS)/VS(ES) of 3:1 was 25% higher than the value of one phase. Results from continuous experiments further demonstrated that VS(CS)/VS(ES) of 3:1 was optimal for hydrogen production with the highest hydrogen yield of 74 mL/gtotal VS added, the balanced nutrient condition with C/N ratio of 1.5 g carbohydrate-COD/gprotein-COD or 11.9 g C/gN might be the main reason for such enhancement. VS(CS)/VS(ES) of 3:1 was also optimal for continuous methane production considering the higher methane yield of 350 mL/gtotal VS added and the lower propionate concentration in the effluent. Copyright © 2010 Elsevier Ltd. All rights reserved.

  18. CHANGES IN THE ANAEROBIC THRESHOLD IN AN ANNUAL CYCLE OF SPORT TRAINING OF YOUNG SOCCER PLAYERS

    PubMed Central

    Andrzejewski, M.; Wieczorek, A.; Barinow-Wojewódzki, A.; Jadczak, Ł.; Adrian, S.; Pietrzak, M.; Wieczorek, S.

    2013-01-01

    The aim of the study was to assess changes in the anaerobic threshold of young soccer players in an annual training cycle. A group of highly trained 15-18 year old players of KKS Lech Poznań were tested. The tests included an annual training macrocycle, and its individual stages resulted from the time structure of the sports training. In order to assess the level of exercise capacities of the players, a field exercise test of increasing intensity was carried out on a soccer pitch. The test made it possible to determine the 4 millimolar lactate threshold (T LA 4 mmol · l-1) on the basis of the lactate concentration in blood [LA], to establish the threshold running speed and the threshold heart rate [HR]. The threshold running speed at the level of the 4 millimolar lactate threshold was established using the two-point form of the equation of a straight line. The obtained indicators of the threshold running speed allowed for precise establishment of effort intensity used in individual training in developing aerobic endurance. In order to test the significance of differences in mean values between four dates of tests, a non-parametric Friedman ANOVA test was used. The significance of differences between consecutive dates of tests was determined using a post-hoc Friedman ANOVA test. The tests showed significant differences in values of selected indicators determined at the anaerobic threshold in various stages of an annual training cycle of young soccer players. The most beneficial changes in terms of the threshold running speed were noted on the fourth date of tests, when the participants had the highest values of 4.01 m · s-1 for older juniors, and 3.80 m · s-1 for younger juniors. This may be indicative of effective application of an individualized programme of training loads and of good preparation of teams for competition in terms of players’ aerobic endurance. PMID:24744480

  19. Changes in the anaerobic threshold in an annual cycle of sport training of young soccer players.

    PubMed

    Sliwowski, R; Andrzejewski, M; Wieczorek, A; Barinow-Wojewódzki, A; Jadczak, L; Adrian, S; Pietrzak, M; Wieczorek, S

    2013-06-01

    The aim of the study was to assess changes in the anaerobic threshold of young soccer players in an annual training cycle. A group of highly trained 15-18 year old players of KKS Lech Poznań were tested. The tests included an annual training macrocycle, and its individual stages resulted from the time structure of the sports training. In order to assess the level of exercise capacities of the players, a field exercise test of increasing intensity was carried out on a soccer pitch. The test made it possible to determine the 4 millimolar lactate threshold (T LA 4 mmol · l(-1)) on the basis of the lactate concentration in blood [LA], to establish the threshold running speed and the threshold heart rate [HR]. The threshold running speed at the level of the 4 millimolar lactate threshold was established using the two-point form of the equation of a straight line. The obtained indicators of the threshold running speed allowed for precise establishment of effort intensity used in individual training in developing aerobic endurance. In order to test the significance of differences in mean values between four dates of tests, a non-parametric Friedman ANOVA test was used. The significance of differences between consecutive dates of tests was determined using a post-hoc Friedman ANOVA test. The tests showed significant differences in values of selected indicators determined at the anaerobic threshold in various stages of an annual training cycle of young soccer players. The most beneficial changes in terms of the threshold running speed were noted on the fourth date of tests, when the participants had the highest values of 4.01 m · s(-1) for older juniors, and 3.80 m · s(-1) for younger juniors. This may be indicative of effective application of an individualized programme of training loads and of good preparation of teams for competition in terms of players' aerobic endurance.

  20. Comparison of the methane production potential and biodegradability of kitchen waste from different sources under mesophilic and thermophilic conditions.

    PubMed

    Yang, Ziyi; Wang, Wen; Zhang, Shuyu; Ma, Zonghu; Anwar, Naveed; Liu, Guangqing; Zhang, Ruihong

    2017-04-01

    The methane production potential of kitchen waste (KW) obtained from different sources was compared through mesophilic and thermophilic anaerobic digestion. The methane yields (MYs) obtained with the same KW sample under different temperatures were similar, whereas the MYs obtained with different samples differed significantly. The highest MY obtained in S7 was 54%-60% higher than the lowest MY in S3. The modified Gompertz model was utilized to simulate the methane production process. The maximum production rate of methane under thermophilic conditions was 2%-86% higher than that under mesophilic conditions. The characteristics of different KW samples were studied. In the distribution of total chemical oxygen demand, the diversity of organic compounds of KW was the most dominant factor that affected the potential MYs of KW. The effect of the C/N and C/P ratios or the concentration of metal ions was insignificant. Two typical methods to calculate the theoretical MY (TMY) were compared, the organic composition method can simulate methane production more precisely than the elemental analysis method. Significant linear correlations were found between TMY org and MYs under mesophilic and thermophilic conditions. The organic composition method can thus be utilized as a fast technique to predict the methane production potential of KW.

  1. Enhanced coproduction of hydrogen and methane from cornstalks by a three-stage anaerobic fermentation process integrated with alkaline hydrolysis.

    PubMed

    Cheng, Xi-Yu; Liu, Chun-Zhao

    2012-01-01

    A three-stage anaerobic fermentation process including H(2) fermentation I, H(2) fermentation II, methane fermentation was developed for the coproduction of hydrogen and methane from cornstalks. Hydrogen production from cornstalks using direct microbial conversion by Clostridium thermocellum 7072 was markedly enhanced in the two-stage thermophilic hydrogen fermentation process integrated with alkaline treatment. The highest total hydrogen yield from cornstalks in the two-stage fermentation process reached 74.4 mL/g-cornstalk. The hydrogen fermentation effluents and alkaline hydrolyzate were further used for methane fermentation by anaerobic granular sludge, and the total methane yield reached 205.8 mL/g-cornstalk. The total energy recovery in the three-stage anaerobic fermentation process integrated with alkaline hydrolysis reached 70.0%. Copyright © 2011 Elsevier Ltd. All rights reserved.

  2. Kinetics of sulfate and hydrogen uptake by the thermophilic sulfate-reducing bacteria Thermodesulfobacterium sp. strain JSP and Thermodesulfovibrio sp. strain R1Ha3

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Sonne-Hansen, J.; Ahring, B.K.; Westermann, P.

    1999-03-01

    Dissimilatory sulfate reduction and methanogenesis are the main terminal processes in the anaerobic food chain. Both the sulfate-reducing bacteria (SRB) and the methane-producing archaea (MPA) use acetate and hydrogen as substrates and, therefore, compete for common electron donors in sulfate-containing natural environments. Due to a higher affinity for the electron donors acetate and hydrogen, SRB outcompete MPA for these compounds whenever sulfate is present in sufficient concentrations. Half-saturation constants (K{sub m}), maximum uptake rates (V{sub max}), and threshold concentrations for sulfate and hydrogen were determined for two thermophilic sulfate-reducing bacteria (SRB) in an incubation system without headspace. K{sub m} valuesmore » determined for the thermophilic SRB were similar to the constants described for mesophilic SRB isolated from environments with low sulfate concentrations.« less

  3. A need for a standardization in anaerobic digestion experiments? Let's get some insight from meta-analysis and multivariate analysis.

    PubMed

    Lavergne, Céline; Jeison, David; Ortega, Valentina; Chamy, Rolando; Donoso-Bravo, Andrés

    2018-09-15

    An important variability in the experimental results in anaerobic digestion lab test has been reported. This study presents a meta-analysis coupled with multivariate analysis aiming to assess the impact of this experimental variability in batch and continuous operation at mesophilic and thermophilic anaerobic digestion of waste activated sludge. An analysis of variance showed that there was no significant difference between mesophilic and thermophilic conditions in both continuous and batch conditions. Concerning the operation mode, the values of methane yield were significantly higher in batch experiment than in continuous reactors. According to the PCA, for both cases, the methane yield is positive correlated to the temperature rises. Interestingly, in the batch experiments, the higher the volatile solids in the substrate was, the lowest was the methane production, which is correlated to experimental flaws when setting up those tests. In continuous mode, unlike the batch test, the methane yield is strongly (positively) correlated to the organic content of the substrate. Experimental standardization, above all, in batch conditions are urgently necessary or move to continuous experiments for reporting results. The modeling can also be a source of disturbance in batch test. Copyright © 2018 Elsevier Ltd. All rights reserved.

  4. Biomass characterization of laboratory-scale thermophilic-mesophilic wastewater treatment processes.

    PubMed

    Suvilampi, J; Lehtomäki, A; Rintala, J

    2006-01-01

    Two thermophilic-mesophilic wastewater treatment processes, one as the combination of the thermophilic activated sludge process (ASP), followed by the mesophilic ASP and the other as thermophilic suspended carrier biofilm process (SCBP), followed by the mesophilic ASP, were used to study sludge characteristics and floc formation. Thermophilic bacteria in both ASP and SCBP were able to form flocs, which were <50 microm in size and had a weak structure and irregular shape. Flocs in both the mesophilic ASPs were larger in size (50-500 microm) and had more compact structures. Filamentous bacteria played an important role in both the thermophilic and mesophilic processes by forming bridges between small flocs. Both thermophilic processes showed a high density of dispersed particles, such as free bacteria. When hydraulic retention time (HRT) was decreased the biofilm was retained in the thermophilic SCBP better than the flocs in the thermophilic ASP. The mesophilic ASPs efficiently removed dispersed particles originating from the thermophilic processes.

  5. Thermoterrabacterium ferrireducens gen. nov., sp. nov., a thermophilic anaerobic dissimilatory Fe(III)-reducing bacterium from a continental hot spring

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Slobodkin, A.; Wiegel, J.; Reysenbach, A.L.

    1997-04-01

    A strain of a thermophilic, anaerobic, dissimilatory, Fe(III)-reducing bacterium, Thermoterrabacterium ferrireducens gen. nov., sp. nov. (type strain JW/AS-Y7{sup T}; DSM 11255), was isolated from hot springs in Yellowstone National Park and New Zealand. The gram-positive-staining cells occurred singly or in pairs as straight to slightly curved rods, 0.3 to 0.4 by 1.6 to 2.7 {mu}m, with rounded ends and exhibited a tumbling motility. Spores were not observed. The temperature range for growth was 50 to 74{degrees}C with an optimum at 65{degrees}C. The pH range for growth at 65{degrees}C was from 5.5 to 7.6, with an optimum at 6.0 to 6.2.more » The organism coupled the oxidation of glycerol to reduction of amorphous Fe(III) oxide or Fe(III) citrate as an electron acceptor. In the presence as well as in the absence of Fe(III) and in the presence of CO{sub 2}, glycerol was metabolized by incomplete oxidation to acetate as the only organic metabolic product; no H{sub 2} was produced during growth. The organism utilized glycerol, lactate, 1,2-propanediol, glycerate, pyruvate, glucose, fructose, mannose, and yeast extract as substrates. In the presence of Fe(III) the bacterium utilized molecular hydrogen. The organism reduced 9,10-anthraquinone-2,6-disulfonic acid, fumarate (to succinate), and thiosulfate (to elemental sulfur) but did not reduce MnO{sub 2}, nitrate, sulfate, sulfite, or elemental sulfur. The G+C content of the DNA was 41 mol% (as determined by high-performance liquid chromatography). The 16S ribosomal DNA sequence analysis placed the isolated strain as a member of a new genus within the gram-type positive Bacillus-Clostridium subphylum.« less

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

    PubMed

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

    2017-12-01

    Microbial community structure in a farm-scale anaerobic digester treating swine manure was investigated during three process events: 1) prolonged starvation, and changes of 2) operating temperature (between meso- and thermophilic) and 3) hydraulic retention time (HRT). Except during the initial period, the digester was dominated by hydrogenotrophic methanogens (HMs). The bacterial community structure significantly shifted with operating temperature and HRT but not with long-term starvation. Clostridiales (26.5-54.4%) and Bacteroidales (2.5-13.7%) became dominant orders in the digester during the period of HM dominance. Abundance of diverse meso- and thermophilic bacteria increased during the same period; many of these species may be H 2 producers, and/or syntrophic acetate oxidizers. Some of these species showed positive correlations with [NH 4 + -N] (p<0.1); this relationship suggests that ammonia was a significant parameter for bacterial selection. The bacterial niche information reported in this study can be useful to understand the ecophysiology of anaerobic digesters treating swine manure that contains high ammonia content. Copyright © 2017 Elsevier B.V. All rights reserved.

  7. Hemicellulose conversion by anaerobic digestion

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Ghosh, S.; Henry, M.P.; Christopher, R.W.

    1982-01-01

    The digestibility of an aquatic biomass (water hyacinth), a land-based biomass (Coastal Bermuda grass), and a biomass-waste blend (a mixture of hyacinth, grass, MSW, and sludge) under various digestion conditions was studied. Anaerobic digestion of hemicellulose consists of the steps of enzymatic hydrolysis of hemicellulose to glucans, mannans, galactans, xylans, and arabans, and then to simple hexose and pentose sugars; production of C/sub 2/ and higher fatty acids from the simple sugars; conversion of higher fatty acids to acetate; and finally, production of methane and CO/sub 2/ from acetate, and CO/sub 2/ and hydrogen. The conversion of hemicellulose was highermore » under mesophilic conditions than those of cellulose or protein for all biomass test feeds, probably because the hemicellulose structure was more vulnerable to enzymatic attack than that of the lignocellulosic component. Cellulose conversion efficiencies at the mesophilic and thermophilic temperatures were about the same. However, hemicellulose was converted at a much lower efficiency than cellulose during thermophilic digestion - a situation that was the reverse of that observed at the mesophilic temperature. Cellulose was utilized in preference to hemicellulose during mesophilic digestion of nitrogen-supplemented Bermuda grass. It was speculated that Bermuda grass cellulose was converted at a higher efficiency than hemicellulose in the presence of external nitrogen because the metabolism of the breakdown product (glucose) of cellulose requires the least investment of enzymes and energy.« less

  8. Thermophilic anaerobic degradation of butyrate by a butyrate-utilizing bacterium in coculture and triculture with methanogenic bacteria.

    PubMed

    Ahring, B K; Westermann, P

    1987-02-01

    We studied syntrophic butyrate degradation in thermophilic mixed cultures containing a butyrate-degrading bacterium isolated in coculture with Methanobacterium thermoautotrophicum or in triculture with M. thermoautotrophicum and the TAM organism, a thermophilic acetate-utilizing methanogenic bacterium. Butyrate was beta-oxidized to acetate with protons as the electron acceptors. Acetate was used concurrently with its production in the triculture. We found a higher butyrate degradation rate in the triculture, in which both hydrogen and acetate were utilized, than in the coculture, in which acetate accumulated. Yeast extract, rumen fluid, and clarified digestor fluid stimulated butyrate degradation, while the effect of Trypticase was less pronounced. Penicillin G, d-cycloserine, and vancomycin caused complete inhibition of butyrate utilization by the cultures. No growth or degradation of butyrate occurred when 2-bromoethanesulfonic acid or chloroform, specific inhibitors of methanogenic bacteria, was added to the cultures and common electron acceptors such as sulfate, nitrate, and fumarate were not used with butyrate as the electron donor. Addition of hydrogen or oxygen to the gas phase immediately stopped growth and butyrate degradation by the cultures. Butyrate was, however, metabolized at approximately the same rate when hydrogen was removed from the cultures and was metabolized at a reduced rate in the cultures previously exposed to hydrogen.

  9. Caloramator quimbayensis sp. nov., an anaerobic, moderately thermophilic bacterium isolated from a terrestrial hot spring.

    PubMed

    Rubiano-Labrador, Carolina; Baena, Sandra; Díaz-Cárdenas, Carolina; Patel, Bharat K C

    2013-04-01

    An anaerobic, moderately thermophilic, terminal-spore-forming bacterium, designated strain USBA A(T), was isolated from a terrestrial hot spring located at an altitude of 2683 m in the Andean region of Colombia (04° 50' 14.0″ N 75° 32' 53.4″ W). Cells of strain USBA A(T) were Gram-stain-positive, straight to slightly curved rods (0.9×2.5 µm), that were arranged singly or in pairs, and were motile by means of flagella. Growth occurred at 37-55 °C and pH 6.0-8.0, with a doubling time of 2 h under the optimal conditions (50 °C and pH 7.0). Glucose fermentation in strain USBA A(T) required yeast extract or peptone (each at 0.2 %, w/v). The novel strain fermented sugars, amino acids, Casamino acids, propanol, propionate, starch and dextrin, but no growth was observed on galactose, lactose, xylose, histidine, serine, threonine, benzoate, butyrate, lactate, pyruvate, succinate, methanol, ethanol, glycerol, casein, gelatin or xylan. The end products of glucose fermentation were formate, acetate, ethanol and lactate. Strain USBA A(T) did not grow autotrophically (with CO2 as carbon source and H2 as electron donor) and did not reduce thiosulfate, sulfate, elemental sulfur, sulfite, vanadium (V) or Fe (III) citrate. Growth of strain USBA A(T) was inhibited by ampicillin, chloramphenicol, kanamycin, penicillin and streptomycin (each at 10 µg ml(-1)). The predominant fatty acids were iso-C15 : 0, C16 : 0 and iso-C17 : 0 and the genomic DNA G+C content was 32.6 mol%. 16S rRNA gene sequence analysis indicated that strain USBA A(T) belonged in the phylum Firmicutes and that its closest relative was Caloramator viterbiensis JW/MS-VS5(T) (95.0 % sequence similarity). A DNA-DNA relatedness value of only 30 % was recorded in hybridization experiments between strain USBA A(T) and Caloramator viterbiensis DSM 13723(T). Based on the phenotypic, chemotaxonomic and phylogenetic evidence and the results of the DNA-DNA hybridization experiments, strain USBA A

  10. The phylogeny of archaebacteria, including novel anaerobic thermoacidophiles in the light of RNA polymerase structure

    NASA Astrophysics Data System (ADS)

    Zillig, Wolfram; Schnabel, Ralf; Tu, Jenn; Stetter, Karl Otto

    1982-05-01

    DNA-dependent RNA polymerases of archaebacteria are distinct from those of eubacteria both in structure and in function. They show similarities to those of the eukaryotic cytoplasm. Extremely thermophilic anaerobic sulfur-respiring archaebacteria isolated from solfataric waters represent four different families, the Thermoproteaceae, the “stiff filaments”, the Desulfurococcaceae and the Thermococcaceae, of a novel order, Thermoproteales. Together with the Sulfolobales, they form the second branch of the urkingdom of the archaebacteria besides that of the methanogens and extreme halophiles. Thermoplasma appears isolated.

  11. Autoheated thermophilic aerobic digestion

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Deeny, K.; Hahn, H.; Leonhard, D.

    1991-10-01

    Autothermal thermophilic aerobic digestion (ATAD) is first and foremost a digestion process, the primary purpose of which is to decompose a portion of the waste organic solids generated from wastewater treatment. As a result of the high operating temperature, digestion is expected to occur within a short time period (6 days) and accomplish a high degree of pathogen reduction. ATAD systems are two-stage aerobic digestion processes that operate under thermophilic temperature conditions (40 to 80C) without supplemental heat. Like composting, the systems rely on the conservation of heat released during digestion itself to attain and sustain the desired operating temperature.more » Typical ATAD systems operate at 55C and may reach temperatures of 60 to 65C in the second-stage reactor. Perhaps because of the high operating temperature, this process has been referred to as Liquid Composting.' Major advantages associated with thermophilic operation include high biological reaction rates and a substantial degree of pathogen reduction.« less

  12. Reduced temperature hydrolysis at 134 °C before thermophilic anaerobic digestion of waste activated sludge at increasing organic load.

    PubMed

    Gianico, A; Braguglia, C M; Cesarini, R; Mininni, G

    2013-09-01

    The performance of thermophilic digestion of waste activated sludge, either untreated or thermal pretreated, was evaluated through semi-continuous tests carried out at organic loading rates in the range of 1-3.7 kg VS/m(3)d. Although the thermal pretreatment at T=134 °C proved to be effective in solubilizing organic matter, no significant gain in organics degradation was observed. However, the digestion of pretreated sludge showed significant soluble COD removal (more than 55%) whereas no removal occurred in control reactors. The lower the initial sludge biodegradability, the higher the efficiency of thermal pretreated digestion was observed, in particular as regards higher biogas and methane production rates with respect to the parallel untreated sludge digestion. Heat balance of the combined thermal hydrolysis/thermophilic digestion process, applied on full-scale scenarios, showed positive values for direct combustion of methane. In case of combined heat and power generation, attractive electric energy recoveries were obtained, with a positive heat balance at high load. Copyright © 2013. Published by Elsevier Ltd.

  13. Caldithrix abyssi gen. nov., sp. nov., a nitrate-reducing, thermophilic, anaerobic bacterium isolated from a Mid-Atlantic Ridge hydrothermal vent, represents a novel bacterial lineage.

    PubMed

    Miroshnichenko, Margarita L; Kostrikina, Nadezhda A; Chernyh, Nikolai A; Pimenov, Nikolai V; Tourova, Tatyana P; Antipov, Alexei N; Spring, Stefan; Stackebrandt, Erko; Bonch-Osmolovskaya, Elizaveta A

    2003-01-01

    A novel, moderately thermophilic, strictly anaerobic, mixotrophic bacterium, designated strain LF13T, was isolated from a deep-sea hydrothermal chimney sample that was collected at a vent site at 14 degrees 45' N, 44 degrees 59' W on the Mid-Atlantic Ridge. Cells were Gram-negative, thin, non-motile rods of variable length. Strain LF13T grew optimally at pH 6.8-7.0 and 60 degrees C with 2.5% (w/v) NaCl. It grew chemo-organoheterotrophically, fermenting proteinaceous substrates, pyruvate and Casamino acids. The strain was able to grow by respiration, utilizing molecular hydrogen (chemolithoheterotrophically) or acetate as electron donors and nitrate as an electron acceptor. Ammonium was formed in the course of denitrification. One-hundred milligrams of yeast extract per litre were required for growth of the strain. The G + C content of the genomic DNA of strain LF13T was 42.5 mol%. Neither 16S rDNA sequence similarity values nor phylogenetic analysis unambiguously related strain LF13T with members of any recognized bacterial phyla. On the basis of 16S rDNA sequence comparisons, and in combination with physiological and morphological traits, a novel genus, Caldithrix, is proposed, with strain LF13T (= DSM 13497T =VKM B-2286T) representing the type species, Caldithrix abyssi.

  14. Were the original eubacteria thermophiles?

    NASA Technical Reports Server (NTRS)

    Achenbach-Richter, L.; Gupta, R.; Stetter, K. O.; Woese, C. R.; Johnson, P. C. (Principal Investigator)

    1987-01-01

    Thermotoga maritima is one of the more unusual eubacteria: It is highly thermophilic, growing at temperatures higher than any other eubacterium; its cell wall appears to have a unique structure and its lipids a unique composition; and the organism is surrounded by a loose-fitting sheath of unknown function. Its phenotypic uniqueness is matched by its phylogenetic position; Thermotoga maritima represents the deepest known branching in the eubacterial line of descent, as measured by ribosomal RNA sequence comparisons. T. maritima also represents the most slowly evolving of eubacterial lineages. The fact that the two deepest branchings in the eubacterial line of descent (the other, the green non-sulfur bacteria and relatives, i.e. Chloroflexus, Thermomicrobium, etc.) are both basically thermophilic and slowly evolving, strongly suggests that all eubacteria have ultimately arisen from a thermophilic ancestor.

  15. Microbial Insight into a Pilot-Scale Enhanced Two-Stage High-Solid Anaerobic Digestion System Treating Waste Activated Sludge.

    PubMed

    Wu, Jing; Cao, Zhiping; Hu, Yuying; Wang, Xiaolu; Wang, Guangqi; Zuo, Jiane; Wang, Kaijun; Qian, Yi

    2017-11-30

    High solid anaerobic digestion (HSAD) is a rapidly developed anaerobic digestion technique for treating municipal sludge, and has been widely used in Europe and Asia. Recently, the enhanced HSAD process with thermal treatment showed its advantages in both methane production and VS reduction. However, the understanding of the microbial community is still poor. This study investigated microbial communities in a pilot enhanced two-stage HSAD system that degraded waste activated sludge at 9% solid content. The system employed process "thermal pre-treatment (TPT) at 70 °C, thermophilic anaerobic digestion (TAD), and mesophilic anaerobic digestion (MAD)". Hydrogenotrophic methanogens Methanothermobacter spp. dominated the system with relative abundance up to about 100% in both TAD and MAD. Syntrophic acetate oxidation (SAO) bacteria were discovered in TAD, and they converted acetate into H₂ and CO₂ to support hydrogenotrophic methanogenesis. The microbial composition and conversion route of this system are derived from the high solid content and protein content in raw sludge, as well as the operational conditions. This study could facilitate the understanding of the enhanced HSAD process, and is of academic and industrial importance.

  16. Thioarsenate Formation Coupled with Anaerobic Arsenite Oxidation by a Sulfate-Reducing Bacterium Isolated from a Hot Spring.

    PubMed

    Wu, Geng; Huang, Liuqin; Jiang, Hongchen; Peng, Yue'e; Guo, Wei; Chen, Ziyu; She, Weiyu; Guo, Qinghai; Dong, Hailiang

    2017-01-01

    Thioarsenates are common arsenic species in sulfidic geothermal waters, yet little is known about their biogeochemical traits. In the present study, a novel sulfate-reducing bacterial strain Desulfotomaculum TC-1 was isolated from a sulfidic hot spring in Tengchong geothermal area, Yunnan Province, China. The arxA gene, encoding anaerobic arsenite oxidase, was successfully amplified from the genome of strain TC-1, indicating it has a potential ability to oxidize arsenite under anaerobic condition. In anaerobic arsenite oxidation experiments inoculated with strain TC-1, a small amount of arsenate was detected in the beginning but became undetectable over longer time. Thioarsenates (AsO 4-x S x 2- with x = 1-4) formed with mono-, di- and tri-thioarsenates being dominant forms. Tetrathioarsenate was only detectable at the end of the experiment. These results suggest that thermophilic microbes might be involved in the formation of thioarsenates and provide a possible explanation for the widespread distribution of thioarsenates in terrestrial geothermal environments.

  17. Thioarsenate Formation Coupled with Anaerobic Arsenite Oxidation by a Sulfate-Reducing Bacterium Isolated from a Hot Spring

    PubMed Central

    Wu, Geng; Huang, Liuqin; Jiang, Hongchen; Peng, Yue’e; Guo, Wei; Chen, Ziyu; She, Weiyu; Guo, Qinghai; Dong, Hailiang

    2017-01-01

    Thioarsenates are common arsenic species in sulfidic geothermal waters, yet little is known about their biogeochemical traits. In the present study, a novel sulfate-reducing bacterial strain Desulfotomaculum TC-1 was isolated from a sulfidic hot spring in Tengchong geothermal area, Yunnan Province, China. The arxA gene, encoding anaerobic arsenite oxidase, was successfully amplified from the genome of strain TC-1, indicating it has a potential ability to oxidize arsenite under anaerobic condition. In anaerobic arsenite oxidation experiments inoculated with strain TC-1, a small amount of arsenate was detected in the beginning but became undetectable over longer time. Thioarsenates (AsO4-xSx2- with x = 1–4) formed with mono-, di- and tri-thioarsenates being dominant forms. Tetrathioarsenate was only detectable at the end of the experiment. These results suggest that thermophilic microbes might be involved in the formation of thioarsenates and provide a possible explanation for the widespread distribution of thioarsenates in terrestrial geothermal environments. PMID:28769902

  18. LC-ESI-MS/MS identification of polar lipids of two thermophilic Anoxybacillus bacteria containing a unique lipid pattern.

    PubMed

    Rezanka, Tomáš; Kambourova, Margarita; Derekova, Anna; Kolouchová, Irena; Sigler, Karel

    2012-07-01

    Phospholipids and glycolipids from two recently described species belonging to the thermophilic genus Anoxybacillus were analyzed by liquid chromatography-electrospray tandem mass spectrometry (LC/ESI-MS/MS). Analysis of total lipids from the facultatively anaerobic A. bogrovensis on a HILIC (Hydrophilic Interaction LIquid Chromatography) column succeeded in separating diacyl- and plasmalogen phospholipids. The LC/ESI-MS/MS analysis of the strict aerobe A. rupiensis revealed the presence of different unique polar lipids, predominantly alanyl-, lysyl-, and glucosyl-phosphatidylglycerols and cardiolipins. Each of the classes of polar lipids was then analyzed by means of the ESI-MS/MS and more than 140 molecular species of six lipid classes from A. bogrovensis and nearly 200 molecular species of nine classes of polar lipids from A. rupiensis were identified. Five classes of unidentified polar lipids were detected in both strains. Plasmalogens were thus determined for the first time in a facultatively anaerobic bacterium, i.e. A. bogrovensis.

  19. Diversity of thermophiles in a Malaysian hot spring determined using 16S rRNA and shotgun metagenome sequencing.

    PubMed

    Chan, Chia Sing; Chan, Kok-Gan; Tay, Yea-Ling; Chua, Yi-Heng; Goh, Kian Mau

    2015-01-01

    The Sungai Klah (SK) hot spring is the second hottest geothermal spring in Malaysia. This hot spring is a shallow, 150-m-long, fast-flowing stream, with temperatures varying from 50 to 110°C and a pH range of 7.0-9.0. Hidden within a wooded area, the SK hot spring is continually fed by plant litter, resulting in a relatively high degree of total organic content (TOC). In this study, a sample taken from the middle of the stream was analyzed at the 16S rRNA V3-V4 region by amplicon metagenome sequencing. Over 35 phyla were detected by analyzing the 16S rRNA data. Firmicutes and Proteobacteria represented approximately 57% of the microbiome. Approximately 70% of the detected thermophiles were strict anaerobes; however, Hydrogenobacter spp., obligate chemolithotrophic thermophiles, represented one of the major taxa. Several thermophilic photosynthetic microorganisms and acidothermophiles were also detected. Most of the phyla identified by 16S rRNA were also found using the shotgun metagenome approaches. The carbon, sulfur, and nitrogen metabolism within the SK hot spring community were evaluated by shotgun metagenome sequencing, and the data revealed diversity in terms of metabolic activity and dynamics. This hot spring has a rich diversified phylogenetic community partly due to its natural environment (plant litter, high TOC, and a shallow stream) and geochemical parameters (broad temperature and pH range). It is speculated that symbiotic relationships occur between the members of the community.

  20. Thermophilic biofiltration of benzene and toluene.

    PubMed

    Cho, Kyung-Suk; Yoo, Sun-Kyung; Ryu, Hee Wook

    2007-12-01

    In the current studies, we characterized the degradation of a hot mixture of benzene and toluene (BT) gases by a thermophilic biofilter using polyurethane as packing material and high-temperature compost as a microbial source. We also examined the effect of supplementing the biofilter with yeast extract (YE). We found that YE substantially enhanced microbial activity in the thermophilic biofilter. The degrading activity of the biofilter supplied with YE was stable during long-term operation (approximately 100 d) without accumulating excess biomass. The maximum elimination capacity (1,650 g x m(-3) h(-1)) in the biofilter supplemented with YE was 3.5 times higher than that in the biofilter without YE (470 g g x m(-3) h(-1)). At similar retention times, the capacity to eliminate BT for the YE-supplemented biofilter was higher than for previously reported mesophilic biofilters. Thus, thermophilic biofiltration can be used to degrade hydrophobic compounds such as a BT mixture. Finally, 16S rDNA polymerase chain reaction-DGGE (PCR-DGGE) fingerprinting revealed that the thermophilic bacteria in the biofilter included Rubrobacter sp. and Mycobacterium sp.

  1. Anaerobic digestion for treatment of stillage from cellulosic bioethanol production.

    PubMed

    Tian, Zhuoli; Mohan, Gayathri Ram; Ingram, Lonnie; Pullammanappallil, Pratap

    2013-09-01

    Thermophilic anaerobic digestion of stillage from a cellulosic ethanol process that uses sugarcane bagasse as feedstock was investigated. A biochemical methane potential (BMP) of 200 ml CH4 at STP (g VS)(-1) was obtained. The whole stillage was separated into two fractions: a fraction retained on 0.5 mm screen called residue and a fraction passing through 0.5 mm screen called filtrate. About 70% of total methane yield of stillage was produced from the filtrate. The filtrate was anaerobically digested in a 15 L semi-continuously fed digester operated for 91 days at HRTs of 21 and 14 days and organic loading rate (OLR) of 1.85 and 2.39 g COD L(-1) d(-1). The methane yield from the stillage from the digester was about 90% of the yield from the BMP assays. The influent soluble COD (sCOD) was reduced from between 35.4 and 38.8 g COD (L(-1)) to between 7.5 and 8 g COD (L(-1)). Copyright © 2013 Elsevier Ltd. All rights reserved.

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

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Franke-Whittle, Ingrid H., E-mail: ingrid.whittle@uibk.ac.at; Walter, Andreas; Ebner, Christian

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

  3. Influence of thermophilic aerobic digestion as a sludge pre-treatment and solids retention time of mesophilic anaerobic digestion on the methane production, sludge digestion and microbial communities in a sequential digestion process.

    PubMed

    Jang, Hyun Min; Cho, Hyun Uk; Park, Sang Kyu; Ha, Jeong Hyub; Park, Jong Moon

    2014-01-01

    In this study, the changes in sludge reduction, methane production and microbial community structures in a process involving two-stage thermophilic aerobic digestion (TAD) and mesophilic anaerobic digestion (MAD) under different solid retention times (SRTs) between 10 and 40 days were investigated. The TAD reactor (RTAD) was operated with a 1-day SRT and the MAD reactor (RMAD) was operated at three different SRTs: 39, 19 and 9 days. For a comparison, control MAD (RCONTROL) was operated at three different SRTs of 40, 20 and 10 days. Our results reveal that the sequential TAD-MAD process has about 42% higher methane production rate (MPR) and 15% higher TCOD removal than those of RCONTROL when the SRT decreased from 40 to 20 days. Denaturing gradient gel electrophoresis (DGGE) and real-time PCR results indicate that RMAD maintained a more diverse bacteria and archaea population compared to RCONTROL, due to the application of the biological TAD pre-treatment process. In RTAD, Ureibacillus thermophiles and Bacterium thermus were the major contributors to the increase in soluble organic matter. In contrast, Methanosaeta concilii, a strictly aceticlastic methanogen, showed the highest population during the operation of overall SRTs in RMAD. Interestingly, as the SRT decreased to 20 days, syntrophic VFA oxidizing bacteria, Clostridium ultunense sp., and a hydrogenotrophic methanogen, Methanobacterium beijingense were detected in RMAD and RCONTROL. Meanwhile, the proportion of archaea to total microbe in RMAD and RCONTROL shows highest values of 10.5 and 6.5% at 20-d SRT operation, respectively. Collectively, these results demonstrate that the increased COD removal and methane production at different SRTs in RMAD might be attributed to the increased synergism among microbial species by improving the hydrolysis of the rate limiting step in sludge with the help of the biological TAD pre-treatment. Copyright © 2013 Elsevier Ltd. All rights reserved.

  4. Studies on the Thermophilic Actinomycetes1

    PubMed Central

    Tendler, M. D.; Burkholder, P. R.

    1961-01-01

    A total of 1,000 isolates of thermophilic actinomycetes representing two genera, Streptomyces and Thermoactinomyces, were studied. Media for cultivation and for physiological studies were designed. Differences between the two genera are noted and taxonomic criteria for the genus Thermoactinomyces are suggested. The importance of the nutritional environment to the thermophilic habit is noted. PMID:13775873

  5. Anaerobic digestion for simultaneous sewage sludge treatment and CO biomethanation: process performance and microbial ecology.

    PubMed

    Luo, Gang; Wang, Wen; Angelidaki, Irini

    2013-09-17

    Syngas is produced by thermal gasification of both nonrenewable and renewable sources including biomass and coal, and it consists mainly of CO, CO2, and H2. In this paper we aim to bioconvert CO in the syngas to CH4. A novel technology for simultaneous sewage sludge treatment and CO biomethanation in an anaerobic reactor was presented. Batch experiments showed that CO was inhibitory to methanogens, but not to bacteria, at CO partial pressure between 0.25 and 1 atm under thermophilic conditions. During anaerobic digestion of sewage sludge supplemented with CO added through a hollow fiber membrane (HFM) module in continuous thermophilic reactors, CO did not inhibit the process even at a pressure as high as 1.58 atm inside the HFM, due to the low dissolved CO concentration in the liquid. Complete consumption of CO was achieved with CO gas retention time of 0.2 d. Results from high-throughput sequencing analysis showed clear differences of the microbial community structures between the samples from liquid and biofilm on the HFM in the reactor with CO addition. Species close to Methanosarcina barkeri and Methanothermobacter thermautotrophicus were the two main archaeal species involved in CO biomethanation. However, the two species were distributed differently in the liquid phase and in the biofilm. Although the carboxidotrophic activities test showed that CO was converted by both archaea and bacteria, the bacterial species responsible for CO conversion are unknown.

  6. Anaerobic Treatment of Municipal Solid Waste and Sludge for Energy Production and Recycling of Nutrients

    NASA Astrophysics Data System (ADS)

    Leinonen, S.

    This volume contains 18 papers presented at a Nordic workshop dealing with application of anaerobic decomposition processes on various types of organic wastes, held at the Siikasalmi Research and Experimental Station of the University of Joensuu on 1-2 Oct. 1992. Subject coverage of the presentations extends from the biochemical and microbiological principles of organic waste processing to descriptions and practical experiences of various types of treatment plants. The theoretical and experimental papers include studies on anaerobic and thermophilic degradation processes, methanogenesis, effects of hydrogen, treatment of chlorinated and phenolic compounds, and process modeling, while the practical examples range from treatment of various types of municipal, industrial, and mining wastes to agricultural and fish farm effluents. The papers provide technical descriptions of several biogas plants in operation. Geographically, the presentations span the Nordic and Baltic countries.

  7. Thermophilic cellobiohydrolase

    DOEpatents

    Sapra, Rajat; Park, Joshua I.; Datta, Supratim; Simmons, Blake A.

    2017-04-18

    The present invention provides for a composition comprising a polypeptide comprising a first amino acid sequence having at least 70% identity with the amino acid sequence of Csac GH5 wherein said first amino acid sequence has a thermostable or thermophilic cellobiohydrolase (CBH) or exoglucanase activity.

  8. Coupling of anaerobic waste treatment to produce protein- and lipid-rich bacterial biomass

    NASA Astrophysics Data System (ADS)

    Steinberg, Lisa M.; Kronyak, Rachel E.; House, Christopher H.

    2017-11-01

    Future long-term manned space missions will require effective recycling of water and nutrients as part of a life support system. Biological waste treatment is less energy intensive than physicochemical treatment methods, yet anaerobic methanogenic waste treatment has been largely avoided due to slow treatment rates and safety issues concerning methane production. However, methane is generated during atmosphere regeneration on the ISS. Here we propose waste treatment via anaerobic digestion followed by methanotrophic growth of Methylococcus capsulatus to produce a protein- and lipid-rich biomass that can be directly consumed, or used to produce other high-protein food sources such as fish. To achieve more rapid methanogenic waste treatment, we built and tested a fixed-film, flow-through, anaerobic reactor to treat an ersatz wastewater. During steady-state operation, the reactor achieved a 97% chemical oxygen demand (COD) removal rate with an organic loading rate of 1740 g d-1 m-3 and a hydraulic retention time of 12.25 d. The reactor was also tested on three occasions by feeding ca. 500 g COD in less than 12 h, representing 50x the daily feeding rate, with COD removal rates ranging from 56-70%, demonstrating the ability of the reactor to respond to overfeeding events. While investigating the storage of treated reactor effluent at a pH of 12, we isolated a strain of Halomonas desiderata capable of acetate degradation under high pH conditions. We then tested the nutritional content of the alkaliphilic Halomonas desiderata strain, as well as the thermophile Thermus aquaticus, as supplemental protein and lipid sources that grow in conditions that should preclude pathogens. The M. capsulatus biomass consisted of 52% protein and 36% lipids, the H. desiderata biomass consisted of 15% protein and 7% lipids, and the Thermus aquaticus biomass consisted of 61% protein and 16% lipids. This work demonstrates the feasibility of rapid waste treatment in a compact reactor design

  9. The isolation and characterization of new C. thermocellum strains and the evaluation of multiple anaerobic digestion systems

    NASA Astrophysics Data System (ADS)

    Lv, Wen

    The overall objective of my research was to improve the efficiencies of bioconversions that produce renewable energy from lignocellulosic biomass. To this end, my studies addressed issues important to two promising strategies: consolidated bioprocessing (CBP) and anaerobic digestion (AD). CBP achieves saccharolytic enzyme production, hydrolysis, and fermentation in a single step and is considered to be the most cost-effective model. Anaerobic bacteria that can be used in CBP are highly desirable. To that end, two thermophilic and cellulolytic bacterial strains were isolated and characterized (Chapter 3). Based on 16S rRNA gene sequence analysis, both strains CS7 and CS8 are closely related to Clostridium thermocellum ATCC 27405. However, they had significantly higher specific cellulase activities and ethanol/acetate ratios than C. thermocellum ATCC 27405. As a result, CS7 and CS8 are two new highly cellulolytic and ethanologenic C. thermocellum strains, with application potentials in research and development of CBP. As some of the most promising AD processes, two temperature-phased AD (TPAD) systems, in comparison with a thermophilic single-stage AD (TSAD) system and a mesophilic two-stage AD (MTAD) system, were studied in treating high-strength dairy cattle manure. The TPAD systems, with the thermophilic digesters acidified (AT-TPAD, Chapter 4) or operated at neutral pH (NT-TPAD, Chapter 5), were optimized at the thermophilic temperature of 50°C and a volume ratio between the thermophilic and the mesophilic digesters of 1:2. Despite similar methane productions, the NT-TPAD system achieved significantly higher volatile solid (VS) removal than the AT-TPAD system and needed no external pH adjustments (Chapter 6). At the same overall OLR, the TSAD system achieved the highest performance, followed by the NT-TPAD and the MTAD systems (Chapter 7). Each digester harbored distinct yet dynamic microbial populations, some of which were significantly correlated or associated

  10. Lebetimonas natsushimae sp. nov., a novel strictly anaerobic, moderately thermophilic chemoautotroph isolated from a deep-sea hydrothermal vent polychaete nest in the Mid-Okinawa Trough.

    PubMed

    Nagata, Ryousuke; Takaki, Yoshihiro; Tame, Akihiro; Nunoura, Takuro; Muto, Hisashi; Mino, Sayaka; Sawayama, Shigeki; Takai, Ken; Nakagawa, Satoshi

    2017-09-01

    A moderately thermophilic, strictly anaerobic, chemoautotrophic bacterium, designated strain HS1857 T , was isolated from a deep-sea hydrothermal vent at the Noho site in the Mid-Okinawa Trough. Strain HS1857 T grew between 35 and 63°C (optimum 55°C), in the presence of 10-55gl -1 NaCl (optimum 25gl -1 ), and pH 5.5-7.1 (optimum 6.4). Growth occurred with molecular hydrogen as the electron donor and elemental sulfur, nitrate, or selenate as the electron acceptors. Formate could serve as an alternative electron donor with nitrate as an electron acceptor. During growth with nitrate as the electron acceptor, strain HS1857 T produced ammonium and formed a biofilm. CO 2 was utilized as the sole carbon source. The G+C content of the genomic DNA was 33.2mol%. Phylogenetic analysis of the 16S rRNA gene sequence indicated that strain HS1857 T is a member of the order Nautiliales, showing a sequence similarity of 95.0% with Lebetimonas acidiphila Pd55 T . The fatty acid composition was similar to that of L. acidiphila, which was dominated by C 18:0 (47.0%) and C 18:1 (23.7%). Based on the genomic, chemotaxonomic, phenotypic characteristics, the name Lebetimonas natsushimae sp. nov., is proposed. The type strain is HS1857 T (=NBRC 112478 T =DSM 104102 T ). Copyright © 2017 Elsevier GmbH. All rights reserved.

  11. Pathogen inactivation in liquid dairy manure during anaerobic and aerobic digestions

    NASA Astrophysics Data System (ADS)

    Biswas, S.; Pandey, P.; Castillo, A. R.; Vaddella, V. K.

    2014-12-01

    Controlling manure-borne pathogens such as E. coli O157:H7, Salmonella spp. and Listeria monocytogenes are crucial for protecting surface and ground water as well as mitigating risks to human health. In California dairy farms, flushing of dairy manure (mainly animal feces and urine) from freestall barns and subsequent liquid-solid manure separation is a common practice for handling animal waste. The liquid manure fraction is generally pumped into the settling ponds and it goes into aerobic and/or anaerobic lagoons for extended period of time. Considering the importance of controlling pathogens in animal waste, the objective of the study was to understand the effects of anaerobic and aerobic digestions on the survival of three human pathogens in animal waste. The pathogen inactivation was assessed at four temperatures (30, 35, 42, and 50 °C), and the relationships between temperature and pathogen decay were estimated. Results showed a steady decrease of E. coli levels in aerobic and anaerobic digestion processes over the time; however, the decay rates varied with pathogens. The effect of temperature on Salmonella spp. and Listeria monocytogenes survival was different than the E. coli survival. In thermophilic temperatures (42 and 50 °C), decay rate was considerable greater compared to the mesophilic temperatures (30 and 35°C). The E. coli log reductions at 50 °C were 2.1 in both aerobic and anaerobic digestions after 13 days of incubation. The Salmonella spp. log reductions at 50 °C were 5.5 in aerobic digestion, and 5.9 in anaerobic digestion. The Listeria monocytogenes log reductions at 50 °C were 5.0 in aerobic digestion, and 5.6 in anaerobic digestion. The log reduction of E. coli, Salmonella spp., and Listeria monocytogens at 30 °C in aerobic environment were 0.1, 4.7, and 5.6, respectively. In anaerobic environment, the corresponding reductions were 0.4, 4.3, and 5.6, respectively. We anticipate that the outcomes of the study will help improving the

  12. Kinetics of autothermal thermophilic aerobic digestion - application and extension of Activated Sludge Model No 1 at thermophilic temperatures.

    PubMed

    Kovács, R; Miháltz, P; Csikor, Zs

    2007-01-01

    The application of an ASM1-based mathematical model for the modeling of autothermal thermophilic aerobic digestion is demonstrated. Based on former experimental results the original ASM1 was extended by the activation of facultative thermophiles from the feed sludge and a new component, the thermophilic biomass was introduced. The resulting model was calibrated in the temperature range of 20-60 degrees C. The temperature dependence of the growth and decay rates in the model is given in terms of the slightly modified Arrhenius and Topiwala-Sinclair equations. The capabilities of the calibrated model in realistic ATAD scenarios are demonstrated with a focus on autothermal properties of ATAD systems at different conditions.

  13. Microbial Insight into a Pilot-Scale Enhanced Two-Stage High-Solid Anaerobic Digestion System Treating Waste Activated Sludge

    PubMed Central

    Wu, Jing; Cao, Zhiping; Hu, Yuying; Wang, Xiaolu; Wang, Guangqi; Zuo, Jiane; Wang, Kaijun; Qian, Yi

    2017-01-01

    High solid anaerobic digestion (HSAD) is a rapidly developed anaerobic digestion technique for treating municipal sludge, and has been widely used in Europe and Asia. Recently, the enhanced HSAD process with thermal treatment showed its advantages in both methane production and VS reduction. However, the understanding of the microbial community is still poor. This study investigated microbial communities in a pilot enhanced two-stage HSAD system that degraded waste activated sludge at 9% solid content. The system employed process “thermal pre-treatment (TPT) at 70 °C, thermophilic anaerobic digestion (TAD), and mesophilic anaerobic digestion (MAD)”. Hydrogenotrophic methanogens Methanothermobacter spp. dominated the system with relative abundance up to about 100% in both TAD and MAD. Syntrophic acetate oxidation (SAO) bacteria were discovered in TAD, and they converted acetate into H2 and CO2 to support hydrogenotrophic methanogenesis. The microbial composition and conversion route of this system are derived from the high solid content and protein content in raw sludge, as well as the operational conditions. This study could facilitate the understanding of the enhanced HSAD process, and is of academic and industrial importance. PMID:29189754

  14. Thermophilic fungi in the new age of fungal taxonomy.

    PubMed

    de Oliveira, Tássio Brito; Gomes, Eleni; Rodrigues, Andre

    2015-01-01

    Thermophilic fungi are of wide interest due to their potential to produce heat-tolerant enzymes for biotechnological processes. However, the taxonomy of such organisms remains obscure, especially given new developments in the nomenclature of fungi. Here, we examine the taxonomy of the thermophilic fungi most commonly used in industry in light of the recent taxonomic changes following the adoption of the International Code of Nomenclature for Algae, Fungi and Plants and also based on the movement One Fungus = One Name. Despite the widespread use of these fungi in applied research, several thermotolerant fungi still remain classified as thermophiles. Furthermore, we found that while some thermophilic fungi have had their genomes sequenced, many taxa still do not have barcode sequences of reference strains available in public databases. This lack of basic information is a limiting factor for the species identification of thermophilic fungi and for metagenomic studies in this field. Based on next-generation sequencing, such studies generate large amounts of data, which may reveal new species of thermophilic fungi in different substrates (composting systems, geothermal areas, piles of plant material). As discussed in this study, there are intrinsic problems associated with this method, considering the actual state of the taxonomy of thermophilic fungi. To overcome such difficulties, the taxonomic classification of this group should move towards standardizing the commonly used species names in industry and to assess the possibility of including new systems for describing species based on environmental sequences.

  15. Strategies for the anaerobic digestion of the organic fraction of municipal solid waste: an overview.

    PubMed

    Hartmann, H; Ahring, B K

    2006-01-01

    Different process strategies for anaerobic digestion of the organic fraction of municipal solid waste (OFMSW) are reviewed weighing high-solids versus low-solids, mesophilic versus thermophilic and single-stage versus multi-stage processes. The influence of different waste characteristics such as composition of biodegradable fractions, C:N ratio and particle size is described. Generally, source sorting of OFMSW and a high content of food waste leads to higher biogas yields than the use of mechanically sorted OFMSW. Thermophilic processes are more efficient than mesophilic processes in terms of higher biogas yields at different organic loading rates (OLR). Highest biogas yields are achieved by means of wet thermophilic processes at OLRs lower than 6 kg-VS x m(-3) d(-1). High-solids processes appear to be relatively more efficient when OLRs higher than 6 kg-VS x m(-3)d(-1) are applied. Multi-stage systems show in some investigations a higher reduction of recalcitrant organic matter compared to single-stage systems, but they are seldom applied in full-scale. An extended cost-benefit calculation shows that the highest overall benefit of the process is achieved at an OLR that is lower and a hydraulic retention time (HRT) that is longer than those values of OLR and HRT, at which the highest biogas production is achieved.

  16. Thermophiles in the genomic era: Biodiversity, science, and applications.

    PubMed

    Urbieta, M Sofía; Donati, Edgardo R; Chan, Kok-Gan; Shahar, Saleha; Sin, Lee Li; Goh, Kian Mau

    2015-11-01

    Thermophiles and hyperthermophiles are present in various regions of the Earth, including volcanic environments, hot springs, mud pots, fumaroles, geysers, coastal thermal springs, and even deep-sea hydrothermal vents. They are also found in man-made environments, such as heated compost facilities, reactors, and spray dryers. Thermophiles, hyperthermophiles, and their bioproducts facilitate various industrial, agricultural, and medicinal applications and offer potential solutions to environmental damages and the demand for biofuels. Intensified efforts to sequence the entire genome of hyperthermophiles and thermophiles are increasing rapidly, as evidenced by the fact that over 120 complete genome sequences of the hyperthermophiles Aquificae, Thermotogae, Crenarchaeota, and Euryarchaeota are now available. In this review, we summarise the major current applications of thermophiles and thermozymes. In addition, emphasis is placed on recent progress in understanding the biodiversity, genomes, transcriptomes, metagenomes, and single-cell sequencing of thermophiles in the genomic era. Copyright © 2015 Elsevier Inc. All rights reserved.

  17. Consolidated bioprocessing method using thermophilic microorganisms

    DOEpatents

    Mielenz, Jonathan Richard

    2016-02-02

    The present invention is directed to a method of converting biomass to biofuel, and particularly to a consolidated bioprocessing method using a co-culture of thermophilic and extremely thermophilic microorganisms which collectively can ferment the hexose and pentose sugars produced by degradation of cellulose and hemicelluloses at high substrate conversion rates. A culture medium therefor is also provided as well as use of the methods to produce and recover cellulosic ethanol.

  18. Diversity of thermophiles in a Malaysian hot spring determined using 16S rRNA and shotgun metagenome sequencing

    PubMed Central

    Chan, Chia Sing; Chan, Kok-Gan; Tay, Yea-Ling; Chua, Yi-Heng; Goh, Kian Mau

    2015-01-01

    The Sungai Klah (SK) hot spring is the second hottest geothermal spring in Malaysia. This hot spring is a shallow, 150-m-long, fast-flowing stream, with temperatures varying from 50 to 110°C and a pH range of 7.0–9.0. Hidden within a wooded area, the SK hot spring is continually fed by plant litter, resulting in a relatively high degree of total organic content (TOC). In this study, a sample taken from the middle of the stream was analyzed at the 16S rRNA V3-V4 region by amplicon metagenome sequencing. Over 35 phyla were detected by analyzing the 16S rRNA data. Firmicutes and Proteobacteria represented approximately 57% of the microbiome. Approximately 70% of the detected thermophiles were strict anaerobes; however, Hydrogenobacter spp., obligate chemolithotrophic thermophiles, represented one of the major taxa. Several thermophilic photosynthetic microorganisms and acidothermophiles were also detected. Most of the phyla identified by 16S rRNA were also found using the shotgun metagenome approaches. The carbon, sulfur, and nitrogen metabolism within the SK hot spring community were evaluated by shotgun metagenome sequencing, and the data revealed diversity in terms of metabolic activity and dynamics. This hot spring has a rich diversified phylogenetic community partly due to its natural environment (plant litter, high TOC, and a shallow stream) and geochemical parameters (broad temperature and pH range). It is speculated that symbiotic relationships occur between the members of the community. PMID:25798135

  19. Optimizing the thermophilic hydrolysis of grass silage in a two-phase anaerobic digestion system.

    PubMed

    Orozco, A M; Nizami, A S; Murphy, J D; Groom, E

    2013-09-01

    Thermophilic hydrolysis of grass silage (GS) at 55 °C with organic loading rates (OLRs) of 6.5, 5, 2.5 and 1.0 kg VS m(-3) days(-1) and hydraulic retention times (HRT) of 10, 6, 4 and 2 days were evaluated in 12 glass bioreactors side by side. The hydrolytic process was measured by variation in pH, volatile solids (VS), VS destruction, soluble chemical oxygen demand (sCOD), hydrolysis and acidification yields. Biological methane potential (BMP) assays were carried out to measure the upper limit for methane production of grass silage with different hydrolytic pretreatments at mesophilic temperature (37 °C). The optimum methane yield of 368 LN CH4 kg(-1) VS was obtained at an OLR of 1 kg VS m(-3)days(-1) and a HRT of 4 days, showing an increase of 30% in the methane potential in comparison to non-hydrolysed GS. Copyright © 2013 Elsevier Ltd. All rights reserved.

  20. Conversion of municipal solid wastes to carboxylic acids by thermophilic fermentation.

    PubMed

    Chan, Wen Ning; Holtzapple, Mark T

    2003-11-01

    The purpose of this research is to generate carboxylic acids from the biodegradable fraction of municipal solid wastes (MSW) and municipal sewage sludge (MSS) by using a thermophilic (55 degrees C), anaerobic, high-solid fermentation. With terrestrial inocula, the highest total carboxylic acid concentration achieved was 20.5 g/L, the highest conversion obtained was 69%, and the highest acetic acid selectivity was 86.4%. Marine inocula were also used to compare against terrestrial sources. Continuum particle distribution modeling (CPDM) was used to predict the final acid product concentrations and substrate conversions at a wide range of liquid residence times (LRT) and volatile solid loading rates (VSLR). "Maps" showing the product concentration and conversion for various LRT and VSLR were generated from CPDM. The predictions were compared to the experimental results. On average, the difference between the predicted and experimental values were 13% for acid concentration and 10% for conversion. CPDM "maps" show that marine inocula produce higher concentrations than terrestrial inocula.

  1. Horse manure as feedstock for anaerobic digestion.

    PubMed

    Hadin, Sa; Eriksson, Ola

    2016-10-01

    Horse keeping is of great economic, social and environmental benefit for society, but causes environmental impacts throughout the whole chain from feed production to manure treatment. According to national statistics, the number of horses in Sweden is continually increasing and is currently approximately 360,000. This in turn leads to increasing amounts of horse manure that have to be managed and treated. Current practices could cause local and global environmental impacts due to poor performance or lack of proper management. Horse manure with its content of nutrients and organic material can however contribute to fertilisation of arable land and recovery of renewable energy following anaerobic digestion. At present anaerobic digestion of horse manure is not a common treatment. In this paper the potential for producing biogas and biofertiliser from horse manure is analysed based on a thorough literature review in combination with mathematical modelling and simulations. Anaerobic digestion was chosen as it has a high degree of resource conservation, both in terms of energy (biogas) and nutrients (digestate). Important factors regarding manure characteristics and operating factors in the biogas plant are identified. Two crucial factors are the type and amount of bedding material used, which has strong implications for feedstock characteristics, and the type of digestion method applied (dry or wet process). Straw and waste paper are identified as the best materials in an energy point of view. While the specific methane yield decreases with a high amount of bedding, the bedding material still makes a positive contribution to the energy balance. Thermophilic digestion increases the methane generation rate and yield, compared with mesophilic digestion, but the total effect is negligible. Copyright © 2016. Published by Elsevier Ltd.

  2. Thermophilic bio-hydrogen production from corn-bran residue pretreated by calcined-lime mud from papermaking process.

    PubMed

    Zhang, Jishi; Zhang, Junjie; Zang, Lihua

    2015-12-01

    This study investigated the use of calcined-lime mud from papermaking process (CLMP) pretreatment to improve fermentative hydrogen yields from corn-bran residue (CBR). CBR samples were pretreated with different concentrations (0-15 g/L) of CLMP at 55°C for 48 h, prior to the thermophilic fermentation with heat-treated anaerobic sludge inoculum. The maximum hydrogen yield (MHY) of 338.91 ml/g-VS was produced from the CBR pretreated with 10 g/L CLMP, with the corresponding lag-phase time of 8.24h. Hydrogen yield increments increased from 27.76% to 48.07%, compared to the control. The CLMP hydrolyzed more cellulose, which provided adequate substrates for hydrogen production. Copyright © 2015 Elsevier Ltd. All rights reserved.

  3. Petrothermobacter organivorans gen. nov., sp. nov., a thermophilic, strictly anaerobic bacterium of the phylum Deferribacteres isolated from a deep subsurface oil reservoir.

    PubMed

    Tamazawa, Satoshi; Mayumi, Daisuke; Mochimaru, Hanako; Sakata, Susumu; Maeda, Haruo; Wakayama, Tatsuki; Ikarashi, Masayuki; Kamagata, Yoichi; Tamaki, Hideyuki

    2017-10-01

    A novel thermophilic, anaerobic, chemoheterotrophic, acetate-oxidizing and iron(III)-, manganese(IV)-, nitrate- and sulfate-reducing bacterium, designated strain ANA T , was isolated from a deep subsurface oil field in Japan (Yabase oil field, Akita Pref.). Cells of strain ANA T were Gram-stain-negative, non-motile, non-spore forming and slightly curved or twisted rods (1.5-5.0 µm long and 0.6-0.7 µm wide). The isolate grew at 25-60 °C (optimum 55 °C) and pH 6.0-8.0 (optimum pH 7.0). The isolate was capable of reducing iron(III), manganese(IV), nitrate and sulfate as an electron acceptor. The isolate utilized a limited range of electron donors such as acetate, lactate, pyruvate and yeast extract for iron reduction. Strain ANA T also used pyruvate, fumarate, succinate, malate, yeast extract and peptone for fermentative growth. The major respiratory quinones were menaquinone-7(H8) and menaquinone-8. The strain contained C18 : 0, iso-C18 : 0 and C16 : 0 as the major cellular fatty acids. The G+C content of the genomic DNA was 34.3 mol%. Phylogenetic analysis based on 16S rRNA gene sequences indicated that strain ANA T was closely related to Calditerrivibrio nitroreducens in the phylum Deferribacteres with low sequence similarities (89.5 %), and formed a distinct clade within the family Deferribacteraceae. In addition, the isolate is the first sulfate-reducing member of the phylum Deferribacteres. Based on phenotypic, chemotaxonomic and phylogenetic properties, a novel genus and species, Petrothermobacter organivorans gen. nov., sp. nov., is proposed for the isolate (type strain=ANA T = NBRC 112621 T =DSM 105015 T ).

  4. Response of a continuous anaerobic digester to temperature transitions: A critical range for restructuring the microbial community structure and function.

    PubMed

    Kim, Jaai; Lee, Changsoo

    2016-02-01

    Temperature is a crucial factor that significantly influences the microbial activity and so the methanation performance of an anaerobic digestion (AD) process. Therefore, how to control the operating temperature for optimal activity of the microbes involved is a key to stable AD. This study examined the response of a continuous anaerobic reactor to a series of temperature shifts over a wide range of 35-65 °C using a dairy-processing byproduct as model wastewater. During the long-term experiment for approximately 16 months, the reactor was subjected to stepwise temperature increases by 5 °C at a fixed HRT of 15 days. The reactor showed stable performance within the temperature range of 35-45 °C, with the methane production rate and yield being maximum at 45 °C (18% and 26% greater, respectively, than at 35 °C). However, the subsequent increase to 50 °C induced a sudden performance deterioration with a complete cessation of methane recovery, indicating that the temperature range between 45 °C and 50 °C had a critical impact on the transition of the reactor's methanogenic activity from mesophilic to thermophilic. This serious process perturbation was associated with a severe restructuring of the reactor microbial community structure, particularly of methanogens, quantitatively as well as qualitatively. Once restored by interrupted feeding for about two months, the reactor maintained fairly stable performance under thermophilic conditions until it was upset again at 65 °C. Interestingly, in contrast to most previous reports, hydrogenotrophs largely dominated the methanogen community at mesophilic temperatures while acetotrophs emerged as a major group at thermophilic temperature. This implies that the primary methanogenesis route of the reactor shifted from hydrogen- to acetate-utilizing pathways with the temperature shifts from mesophilic to thermophilic temperatures. Our observations suggest that a mesophilic digester may not need to be cooled at up

  5. Thermophilic anaerobic co-digestion of garbage, screened swine and dairy cattle manure.

    PubMed

    Liu, Kai; Tang, Yue-Qin; Matsui, Toru; Morimura, Shigeru; Wu, Xiao-Lei; Kida, Kenji

    2009-01-01

    Methane fermentation characteristics of garbage, swine manure (SM), dairy cattle manure (DCM) and mixtures of these wastes were studied. SM and DCM showed much lower volatile total solid (VTS) digestion efficiencies and methane yield than those of garbage. VTS digestion efficiency of SM was significantly increased when it was co-digested with garbage (Garbage: SM=1:1). Co-digestion of garbage, SM and DCM with respect to the relative quantity of each waste discharged in the Kikuchi (1: 16: 27) and Aso (1: 19: 12) areas indicated that co-digestion with garbage would improve the digestion characteristic of SM and DCM as far as the ratio of DCM in the wastes was maintained below a certain level. When the mixed waste (Garbage: SM: DCM=1:19:12) was treated using a thermophilic UAF reactor, methanogens responsible for the methane production were Methanoculleus and Methanosarcina species. Bacterial species in the phylum Firmicutes were dominant bacteria responsible for the digestion of these wastes. As the percentage of garbage in the mixed wastes used in this study was low (2-3%) and the digestion efficiency of DCM was obviously improved, the co-digestion of SM and DCM with limited garbage was a prospective method to treat the livestock waste effectively and was an attractive alternative technology for the construction of a sustainable environment and society in stock raising area.

  6. Anaerobic co-digestion of municipal organic wastes and pre-treatment to enhance biogas production from waste.

    PubMed

    Li, Chenxi; Champagne, Pascale; Anderson, Bruce C

    2014-01-01

    Co-digestion and pre-treatment have been recognized as effective, low-cost and commercially viable approaches to reduce anaerobic digestion process limitations and improve biogas yields. In our previous batch-scale study, fat, oil, and grease (FOG) was investigated as a suitable potential co-substrate, and thermo-chemical pre-treatment (TCPT) at pH = 10 and 55 °C improved CH4 production from FOG co-digestions. In this project, co-digestions with FOG were studied in bench-scale two-stage thermophilic semi-continuous flow co-digesters with suitable TCPT (pH = 10, 55 °C). Overall, a 25.14 ± 2.14 L/d (70.2 ± 1.4% CH4) biogas production was obtained, which was higher than in the two-stage system without pre-treatment. The results could provide valuable fundamental information to support full-scale investigations of anaerobic co-digestion of municipal organic wastes.

  7. Evaluation of biogas production potential by dry anaerobic digestion of switchgrass--animal manure mixtures.

    PubMed

    Ahn, H K; Smith, M C; Kondrad, S L; White, J W

    2010-02-01

    Anaerobic digestion is a biological method used to convert organic wastes into a stable product for land application with reduced environmental impacts. The biogas produced can be used as an alternative renewable energy source. Dry anaerobic digestion [>15% total solid (TS)] has an advantage over wet digestion (<10% TS) because it allows for the use of a smaller volume of reactor and because it reduces wastewater production. In addition, it produces a fertilizer that is easier to transport. Performance of anaerobic digestion of animal manure-switchgrass mixture was evaluated under dry (15% TS) and thermophilic conditions (55 degrees C). Three different mixtures of animal manure (swine, poultry, and dairy) and switchgrass were digested using batch-operated 1-L reactors. The swine manure test units showed 52.9% volatile solids (VS) removal during the 62-day trial, while dairy and poultry manure test units showed 9.3% and 20.2%, respectively. Over the 62 day digestion, the swine manure test units yielded the highest amount of methane 0.337 L CH4/g VS, while the dairy and poultry manure test units showed very poor methane yield 0.028 L CH4/g VS and 0.002 L CH4/g VS, respectively. Although dairy and poultry manure performed poorly, they may still have high potential as biomass for dry anaerobic digestion if appropriate designs are developed to prevent significant volatile fatty acid (VFA) accumulation and pH drop.

  8. An immunological assay for detection and enumeration of thermophilic biomining microorganisms.

    PubMed

    Amaro, A M; Hallberg, K B; Lindström, E B; Jerez, C A

    1994-09-01

    A specific, fast, and sensitive nonradioactive immunobinding assay for the detection and enumeration of the moderate thermophile Thiobacillus caldus and the thermophilic archaeon Sulfolobus acidocaldarius was developed. It employs enhanced chemiluminescence or peroxidase-conjugated immunoglobulins in a dot or slot blotting system and is very convenient for monitoring thermophilic bioleaching microorganisms in effluents from industrial bioleaching processes.

  9. Hydrophobic environment is a key factor for the stability of thermophilic proteins.

    PubMed

    Gromiha, M Michael; Pathak, Manish C; Saraboji, Kadhirvel; Ortlund, Eric A; Gaucher, Eric A

    2013-04-01

    The stability of thermophilic proteins has been viewed from different perspectives and there is yet no unified principle to understand this stability. It would be valuable to reveal the most important interactions for designing thermostable proteins for such applications as industrial protein engineering. In this work, we have systematically analyzed the importance of various interactions by computing different parameters such as surrounding hydrophobicity, inter-residue interactions, ion-pairs and hydrogen bonds. The importance of each interaction has been determined by its predicted relative contribution in thermophiles versus the same contribution in mesophilic homologues based on a dataset of 373 protein families. We predict that hydrophobic environment is the major factor for the stability of thermophilic proteins and found that 80% of thermophilic proteins analyzed showed higher hydrophobicity than their mesophilic counterparts. Ion pairs, hydrogen bonds, and interaction energy are also important and favored in 68%, 50%, and 62% of thermophilic proteins, respectively. Interestingly, thermophilic proteins with decreased hydrophobic environments display a greater number of hydrogen bonds and/or ion pairs. The systematic elimination of mesophilic proteins based on surrounding hydrophobicity, interaction energy, and ion pairs/hydrogen bonds, led to correctly identifying 95% of the thermophilic proteins in our analyses. Our analysis was also applied to another, more refined set of 102 thermophilic-mesophilic pairs, which again identified hydrophobicity as a dominant property in 71% of the thermophilic proteins. Further, the notion of surrounding hydrophobicity, which characterizes the hydrophobic behavior of residues in a protein environment, has been applied to the three-dimensional structures of elongation factor-Tu proteins and we found that the thermophilic proteins are enriched with a hydrophobic environment. The results obtained in this work highlight the

  10. Psychrophilic dry anaerobic digestion of dairy cow feces: Long-term operation

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Massé, Daniel I., E-mail: Daniel.masse@agr.gc.ca; Cata Saady, Noori M.

    2015-02-15

    Highlights: • Psychrophilic dry anaerobic digestion (PDAD) of cow feces (CF) is feasible. • PDAD of CF is as efficient as mesophilic and thermophilic AD at TCL 21 days. • CF (13–16% TS at OLR 5.0 g TCOD{sub fed} kg{sup −1} inoculum d{sup −1}) yielded 222 ± 27 {sub N}L CH{sub 4} kg{sup −1} VS fed. - Abstract: This paper reports experimental results which demonstrate psychrophilic dry anaerobic digestion of cow feces during long-term operation in sequence batch reactor. Cow feces (13–16% total solids) has been anaerobically digested in 12 successive cycles (252 days) at 21 days treatment cycle lengthmore » (TCL) and temperature of 20 °C using psychrotrophic anaerobic mixed culture. An average specific methane yield (SMY) of 184.9 ± 24.0, 189.9 ± 27.3, and 222 ± 27.7 {sub N}L CH{sub 4} kg{sup −1} of VS fed has been achieved at an organic loading rate of 3.0, 4.0, and 5.0 g TCOD kg{sup −1} inoculum d{sup −1} and TCL of 21 days, respectively. The corresponding substrate to inoculum ratio (SIR) was 0.39 ± 0.06, 0.48 ± .02, 0.53 ± 0.05, respectively. Average methane production rate of 10 ± 1.4 {sub N}L CH{sub 4} kg{sup −1} VS fed d{sup −1} has been obtained. The low concentration of volatile fatty acids indicated that hydrolysis was the reaction limiting step.« less

  11. Coupling of anaerobic waste treatment to produce protein- and lipid-rich bacterial biomass.

    PubMed

    Steinberg, Lisa M; Kronyak, Rachel E; House, Christopher H

    2017-11-01

    Future long-term manned space missions will require effective recycling of water and nutrients as part of a life support system. Biological waste treatment is less energy intensive than physicochemical treatment methods, yet anaerobic methanogenic waste treatment has been largely avoided due to slow treatment rates and safety issues concerning methane production. However, methane is generated during atmosphere regeneration on the ISS. Here we propose waste treatment via anaerobic digestion followed by methanotrophic growth of Methylococcus capsulatus to produce a protein- and lipid-rich biomass that can be directly consumed, or used to produce other high-protein food sources such as fish. To achieve more rapid methanogenic waste treatment, we built and tested a fixed-film, flow-through, anaerobic reactor to treat an ersatz wastewater. During steady-state operation, the reactor achieved a 97% chemical oxygen demand (COD) removal rate with an organic loading rate of 1740 g d -1  m -3 and a hydraulic retention time of 12.25 d. The reactor was also tested on three occasions by feeding ca. 500 g COD in less than 12 h, representing 50x the daily feeding rate, with COD removal rates ranging from 56-70%, demonstrating the ability of the reactor to respond to overfeeding events. While investigating the storage of treated reactor effluent at a pH of 12, we isolated a strain of Halomonas desiderata capable of acetate degradation under high pH conditions. We then tested the nutritional content of the alkaliphilic Halomonas desiderata strain, as well as the thermophile Thermus aquaticus, as supplemental protein and lipid sources that grow in conditions that should preclude pathogens. The M. capsulatus biomass consisted of 52% protein and 36% lipids, the H. desiderata biomass consisted of 15% protein and 7% lipids, and the Thermus aquaticus biomass consisted of 61% protein and 16% lipids. This work demonstrates the feasibility of rapid waste treatment in a compact

  12. Microbial population dynamics during startup of a full-scale anaerobic digester treating industrial food waste in Kyoto eco-energy project.

    PubMed

    Ike, Michihiko; Inoue, Daisuke; Miyano, Tomoki; Liu, Tong Tong; Sei, Kazunari; Soda, Satoshi; Kadoshin, Shiro

    2010-06-01

    The microbial community in a full-scale anaerobic digester (2300m3) treating industrial food waste in the Kyoto Eco-Energy Project was analyzed using terminal restriction fragment length polymorphism for eubacterial and archaeal 16S rRNA genes. Both thermophilic and mesophilic sludge of treated swine waste were seeded to the digestion tank. During the 150-day startup period, coffee grounds as a main food waste, along with potato, kelp and boiled beans, tofu, bean curd lees, and deep-fried bean curd were fed to the digestion process step-by-step (max. 40t/d). Finally, the methane yield reached 360m3/t-feed with 40days' retention time, although temporary accumulation of propionate was observed. Eubacterial communities that formed in the thermophilic digestion tank differed greatly from both thermophilic and mesophilic types of seed sludge. Results suggest that the Actinomyces/Thermomonospora and Ralstonia/Shewanella were contributors for hydrolyzation and degradation of food waste into volatile fatty acids. Acetate-utilizing methanogens, Methanosaeta, were dominant in seed sludges of both types, but they decreased drastically during processing in the digestion tank. Methanosarcina and Methanobrevibacter/Methanobacterium were, respectively, possible main contributors for methane production from acetate and H2 plus CO2. Copyright 2010 Elsevier Ltd. All rights reserved.

  13. Anaerobic wastewater treatment and membrane filtration: a one night stand or a sustainable relationship?

    PubMed

    Jeison, D; van Lier, J B

    2008-01-01

    Several anaerobic membrane bioreactors (AnMBR) were operated, under various conditions, applying different reactor configurations. Applicable fluxes were strongly determined by the physical properties of the sludge present in the reactors. Results show that particle size is a key determining factor for the attainable fluxes. Under thermophilic conditions, small sludge particle size was observed, resulting in low critical fluxes reaching 6-7 L/m2h for the submerged configuration and acidified substrate. In contrast, under mesophilic conditions critical fluxes of 20 L/m2h were obtained. The acidification level also showed a strong effect. Under thermophilic conditions, the presence of a significant fraction of non-acidified organic matter induced the growth of suspended acidogenic biomass that seriously affected the applicable fluxes, both in submerged and side-stream configurations. Under all conditions tested cake formation showed to be the limiting factor determining the applicable fluxes. Only low levels of irreversible fouling were observed. Due to technical and economical considerations, most interesting perspectives for the application of AnMBR are expected with the treatment of high-strength particulate wastewaters, and with extreme wastewaters characterised by high temperature, salinity, etc.

  14. Microbial community dynamics in anaerobic bioreactors and algal tanks treating piggery wastewater.

    PubMed

    Patil, Sayali S; Kumar, Martin S; Ball, Andrew S

    2010-06-01

    Integrated biosystem is becoming a major aspect of wastewater management practice. Microbial communities in piggery wastewater sampled from anaerobic (thermophilic and mesophilic) and aerobic digesters (algal tanks) during waste remediation were analyzed by culture-independent techniques based on polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE). The use of Muyzer's 314F-GC, 518R bacterial primers, and archaeal A934F, 1309R primers followed by partial 16s rDNA sequence analysis of the main bands from DGGE revealed the presence of unknown and as yet uncultured microorganisms but also showed functional and ecologically significant denitrifying, acetogenic bacteria along with autotrophic, hydrogenotrophic, and acetoclastic methanogen archaea. Thermophilic digesters were dominated by gamma-Proteobacteria, Methanothermobacter sp., while mesophilic digesters showed dominance by Firmicutes, uncultured bacteria, Methanosarcina, and Methanoculleus genera. Under aerobic conditions within algal tanks, pH rose from 7.17 to 9.32, with a significant decrease in total ammonia nitrogen, chemical oxygen demand, and soluble phosphorus levels. PCR-DGGE proved a useful tool for investigating the dynamics of microbial community in the bio-processing of piggery wastewater. Knowledge of the microbial communities involved in digestion of piggery wastewater will allow optimization of integrated biosystem by removing the main pollutants like inorganic ammonium-nitrogen, phosphorus, and pathogens from intensive farming system.

  15. An Immunological Assay for Detection and Enumeration of Thermophilic Biomining Microorganisms

    PubMed Central

    Amaro, Ana M.; Hallberg, Kevin B.; Lindström, E. Börje; Jerez, Carlos A.

    1994-01-01

    A specific, fast, and sensitive nonradioactive immunobinding assay for the detection and enumeration of the moderate thermophile Thiobacillus caldus and the thermophilic archaeon Sulfolobus acidocaldarius was developed. It employs enhanced chemiluminescence or peroxidase-conjugated immunoglobulins in a dot or slot blotting system and is very convenient for monitoring thermophilic bioleaching microorganisms in effluents from industrial bioleaching processes. Images PMID:16349398

  16. Assessment of Two Solid Anaerobic Digestate Soil Amendments for Effects on Soil Quality and Biosolarization Efficacy.

    PubMed

    Fernández-Bayo, Jesús D; Achmon, Yigal; Harrold, Duff R; McCurry, Dlinka G; Hernandez, Katie; Dahlquist-Willard, Ruth M; Stapleton, James J; VanderGheynst, Jean S; Simmons, Christopher W

    2017-05-03

    Anaerobic digestion is an organic waste bioconversion process that produces biofuel and digestates. Digestates have potential to be applied as soil amendment to improve properties for crop production including phytonutrient content and pest load. Our objective was to assess the impact of solid anaerobic digestates on weed seed inactivation and soil quality upon soil biosolarization (a pest control technique that combines solar heating and amendment-induced microbial activity). Two solid digestates from thermophilic (TD) and mesophilic (MD) digesters were tested. The solarized TD-amended samples presented significantly higher mortality of Brassica nigra (71%, P = 0.032) than its equivalent incubated at room temperature. However, biosolarization with digestate amendment led to decreased weed seed mortality in certain treatments. The plant-available water, total C, and extractable P and K were significantly increased (P < 0.05) in the incubated amended soils. The results confirm the potential of digestates as beneficial soil amendments. Further studies are needed to elucidate the impacts of digestate stability on biosolarization efficacy and soil properties.

  17. Instability diagnosis and syntrophic acetate oxidation during thermophilic digestion of vegetable waste.

    PubMed

    Li, Dong; Ran, Yi; Chen, Lin; Cao, Qin; Li, Zhidong; Liu, Xiaofeng

    2018-08-01

    Effective process monitoring and instability diagnosis are important for stable anaerobic digestion (AD) of vegetable waste (VW). In order to evaluate the performance of thermophilic digestion of VW, to make early diagnosis for instability after organic overload, and to reveal the dynamics of microbial community under different running states, thermophilic AD of VW was carried out under improved organic loading rates (OLR) of 0.5-2.5 g volatile solid (VS)/(L ∙ d) in this study. Gaseous parameters including volumetric methane production rate (VMPR), CH 4 , CO 2 , and H 2 concentrations, and liquid parameters including pH, oxidation-reduction potential, volatile fatty acid (VFA), and total alkalinity (TA), bicarbonate alkalinity (BA), intermediate alkalinity (IA), and ammonia, were monitored. The coupling parameters, such as the CH 4 /CO 2 , VFA/BA, and BA/TA ratios were also used to evaluate stability. The dynamics of syntrophic acetate-oxidizing bacteria (SAOB), acetoclastic methanogens (AM), and hydrogenotrophic methanogens (HM) were analyzed by high-throughput sequencing. The main methanogenic bacteria were HM (Methanothermobacter) during the start-up period of OLR 0.5 gVS/(L ∙ d), while they were AM (Methanosarcina) during the stable period of OLR of 1.0 gVS/(L ∙ d). The VMPR of stable period was about 0.29 L/(L · d) with total VFA concentration below 100 mg/L, CH 4 /CO 2  > 1.3, and BA/TA>0.9. The first instability due to the accumulation of VFA and self-recovery due to syntrophic acetate oxidation occurred at an OLR of 1.5 gVS/(L ∙ d). The syntrophic acetate-oxidizing bacteria probably belong to genus S1 (family Thermotogaceae). The digestion failed at an OLR of 2.0 g VS/(L · d). H 2 was only detected during collapsed period instead of instable period. The total ammonia nitrogen loss and bicarbonate alkalinity (BA) reduction were the primary causes for the instability of AD of VW without effluent recirculation. Compared with single

  18. Dark fermentation of complex waste biomass for biohydrogen production by pretreated thermophilic anaerobic digestate.

    PubMed

    Ghimire, Anish; Frunzo, Luigi; Pontoni, Ludovico; d'Antonio, Giuseppe; Lens, Piet N L; Esposito, Giovanni; Pirozzi, Francesco

    2015-04-01

    The Biohydrogen Potential (BHP) of six different types of waste biomass typical for the Campania Region (Italy) was investigated. Anaerobic sludge pre-treated with the specific methanogenic inhibitor sodium 2-bromoethanesulfonic acid (BESA) was used as seed inoculum. The BESA pre-treatment yielded the highest BHP in BHP tests carried out with pre-treated anaerobic sludge using potato and pumpkin waste as the substrates, in comparison with aeration or heat shock pre-treatment. The BHP tests carried out with different complex waste biomass showed average BHP values in a decreasing order from potato and pumpkin wastes (171.1 ± 7.3 ml H2/g VS) to buffalo manure (135.6 ± 4.1 ml H2/g VS), dried blood (slaughter house waste, 87.6 ± 4.1 ml H2/g VS), fennel waste (58.1 ± 29.8 ml H2/g VS), olive pomace (54.9 ± 5.4 ml H2/g VS) and olive mill wastewater (46.0 ± 15.6 ml H2/g VS). The digestate was analyzed for major soluble metabolites to elucidate the different biochemical pathways in the BHP tests. These showed the H2 was produced via mixed type fermentation pathways. Copyright © 2015 Elsevier Ltd. All rights reserved.

  19. Methane production by anaerobic digestion of Bermuda grass

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Klass, D.L.; Ghosh, S.

    1979-01-01

    Bermuda grass (Cynodon dactylon) is one of the high-yield warm-season grasses that has been suggested as a promising raw material for conversion to methane. Experimental work performed with laboratory digesters to study the anaerobic digestion of Coastal Bermuda grass harvested in Louisiana and having a C/N ratio of 24 is described. Methane yields of about 1.9 SCF/lb of volatile solids( VS) added were observed under conventional mesophilic high-rate conditions. When supplemental nitrogen additions were made, the yields increased up to 3.5 SCF/lb of VS added indicating that the nitrogen content of the grass examined was insufficient to sustain high-rate digestionmore » at the higher yield level. Thermophilic digestion with supplemental nitrogen additions afforded methane yields of about 2.7 SCF/lb VS added. Carbon and energy balances were calculated and the relative biodegradabilities of the organics were estimated.« less

  20. Membrane lipid patterns typify distinct anaerobic methanotrophic consortia

    PubMed Central

    Blumenberg, Martin; Seifert, Richard; Reitner, Joachim; Pape, Thomas; Michaelis, Walter

    2004-01-01

    The anaerobic oxidation of methane (AOM) is one of the major sinks of this substantial greenhouse gas in marine environments. Recent investigations have shown that diverse communities of anaerobic archaea and sulfate-reducing bacteria are involved in AOM. Most of the relevant archaea are assigned to two distinct phylogenetic clusters, ANME-1 and ANME-2. A suite of specific 13C-depleted lipids demonstrating the presence of consortia mediating AOM in fossil and recent environments has been established. Here we report on substantial differences in the lipid composition of microbial consortia sampled from distinct compartments of AOM-driven carbonate reefs growing in the northwestern Black Sea. Communities in which the dominant archaea are from the ANME-1 cluster yield internally cyclized tetraether lipids typical of thermophiles. Those in which ANME-2 archaea are dominant yield sn-2-hydroxyarchaeol accompanied by crocetane and crocetenes. The bacterial lipids from these communities are also distinct even though the sulfate-reducing bacteria all belong to the Desulfosarcina/Desulfococcus group. Nonisoprenoidal glycerol diethers are predominantly associated with ANME-1-dominated communities. Communities with ANME-2 yield mainly conventional, ester-linked diglycerides. ANME-1 archaea and associated sulfate-reducing bacteria seem to be enabled to use low concentrations of methane and to grow within a broad range of temperatures. Our results offer a tool for the study of recent and especially of fossil methane environments. PMID:15258285

  1. Anaerobic digestion of orange peel in a semi-continuous pilot plant: An environmentally sound way of citrus waste management in agro-ecosystems.

    PubMed

    Zema, Demetrio A; Fòlino, Adele; Zappia, Giovanni; Calabrò, Paolo S; Tamburino, Vincenzo; Zimbone, Santo Marcello

    2018-07-15

    The management of residues of citrus processing involves economic and environmental problems. In particular, the uncontrolled disposal of citrus processing waste near production sites can have heavy impacts on air, soil, surface water bodies and groundwater. Anaerobic digestion has been proposed as a viable alternative for citrus waste valorisation, if some problems, linked to the biochemical processes, are overcome. Although many experimental tests have studied the inhibitory effects of the high essential oil content of orange peel on biomethanisation processes, fewer experiences have been carried out in continuous or semi-continuous pilot digesters, more similar to the full-scale biogas plants, using real orange peel. This study has evaluated the methane production through anaerobic digestion of industrial orange peel using a pilot plant (84L) with semi-continuous feeding at increasing Organic Loading Rates (OLR) and essential oil (EO) supply rates (EOsr) until the complete process inhibition. Under mesophilic conditions, the highest daily specific methane yield was achieved at OLR of 1.0g TVS L -1 d -1 and EOsr of 47.6mgL -1 d -1 . Partial inhibition of the anaerobic digestion was detected at OLR and EOsr of 1.98g TVS L -1 d -1 and 88.1mgL -1 d -1 , respectively and the process irreversibly stopped when OLR and EOsr reached 2.5g TVS L -1 d -1 and 111.2mgL -1 d -1 , respectively. Under thermophilic conditions, the cumulative methane production (0.12Lg TVS -1 ) was about 25% of that under mesophilic conditions (0.46Lg TVS -1 ). The thermophilic digestion was completely inhibited at lower OLR (1.98g TVS L -1 d -1 ) and EOsr (88.1mgL -1 d -1 ) compared to mesophilic conditions. This study confirmed the suitability of anaerobic digestion of orange peel for biomethane production (provided that the right management of the process is set), in view of an environmentally sound way of agricultural residues management in agro-ecosystems. Copyright © 2018 Elsevier B.V. All

  2. Enrichment and detection of microorganisms involved in direct and indirect methanogenesis from methanol in an anaerobic thermophilic bioreactor.

    PubMed

    Roest, Kees; Altinbas, Mahmut; Paulo, Paula L; Heilig, H G H J; Akkermans, Antoon D L; Smidt, Hauke; de Vos, Willem M; Stams, Alfons J M

    2005-10-01

    To gain insight into the microorganisms involved in direct and indirect methane formation from methanol in a laboratory-scale thermophilic (55 degrees C) methanogenic bioreactor, reactor sludge was disrupted and serial dilutions were incubated in specific growth media containing methanol and possible intermediates of methanol degradation as substrates. With methanol, growth was observed up to a dilution of 10(8). However, when Methanothermobacter thermoautotrophicus strain Z245 was added for H2 removal, growth was observed up to a 10(10)-fold dilution. With H2/CO2 and acetate, growth was observed up to dilutions of 10(9) and 10(4), respectively. Dominant microorganisms in the different dilutions were identified by 16S rRNA-gene diversity and sequence analysis. Furthermore, dilution polymerase chain reaction (PCR) revealed a similar relative abundance of Archaea and Bacteria in all investigated samples, except in enrichment with acetate, which contained 100 times less archaeal DNA than bacterial DNA. The most abundant bacteria in the culture with methanol and strain Z245 were most closely related to Moorella glycerini. Thermodesulfovibrio relatives were found with high sequence similarity in the H2/CO2 enrichment, but also in the original laboratory-scale bioreactor sludge. Methanothermobacter thermoautotrophicus strains were the most abundant hydrogenotrophic archaea in the H2/CO2 enrichment. The dominant methanol-utilizing methanogen, which was present in the 10(8)-dilution, was most closely related to Methanomethylovorans hollandica. Compared to direct methanogenesis, results of this study indicate that syntrophic, interspecies hydrogen transfer-dependent methanol conversion is equally important in the thermophilic bioreactor, confirming previous findings with labeled substrates and specific inhibitors.

  3. Deletion of the hfsB gene increases ethanol production in Thermoanaerobacterium saccharolyticum and several other thermophilic anaerobic bacteria.

    PubMed

    Eminoğlu, Ayşenur; Murphy, Sean Jean-Loup; Maloney, Marybeth; Lanahan, Anthony; Giannone, Richard J; Hettich, Robert L; Tripathi, Shital A; Beldüz, Ali Osman; Lynd, Lee R; Olson, Daniel G

    2017-01-01

    With the discovery of interspecies hydrogen transfer in the late 1960s (Bryant et al. in Arch Microbiol 59:20-31, 1967), it was shown that reducing the partial pressure of hydrogen could cause mixed acid fermenting organisms to produce acetate at the expense of ethanol. Hydrogen and ethanol are both more reduced than glucose. Thus there is a tradeoff between production of these compounds imposed by electron balancing requirements; however, the mechanism is not fully known. Deletion of the hfsA or B subunits resulted in a roughly 1.8-fold increase in ethanol yield. The increase in ethanol production appears to be associated with an increase in alcohol dehydrogenase activity, which appears to be due, at least in part, to increased expression of the adhE gene, and may suggest a regulatory linkage between hfsB and adhE . We studied this system most intensively in the organism Thermoanaerobacterium saccharolyticum ; however, deletion of hfsB also increases ethanol production in other thermophilic bacteria suggesting that this could be used as a general technique for engineering thermophilic bacteria for improved ethanol production in organisms with hfs -type hydrogenases. Since its discovery by Shaw et al. (JAMA 191:6457-64, 2009), the hfs hydrogenase has been suspected to act as a regulator due to the presence of a PAS domain. We provide additional support for the presence of a regulatory phenomenon. In addition, we find a practical application for this scientific insight, namely increasing ethanol yield in strains that are of interest for ethanol production from cellulose or hemicellulose. In two of these organisms ( T. xylanolyticum and T. thermosaccharolyticum ), the ethanol yields are the highest reported to date.

  4. Thermophilic xylanases: from bench to bottle.

    PubMed

    Basit, Abdul; Liu, Junquan; Rahim, Kashif; Jiang, Wei; Lou, Huiqiang

    2018-01-17

    Lignocellulosic biomass is a valuable raw material. As technology has evolved, industrial interest in new ways to take advantage of this raw material has grown. Biomass is treated with different microbial cells or enzymes under ideal industrial conditions to produce the desired products. Xylanases are the key enzymes that degrade the xylosidic linkages in the xylan backbone of the biomass, and commercial enzymes are categorized into different glycoside hydrolase families. Thermophilic microorganisms are excellent sources of industrially relevant thermostable enzymes that can withstand the harsh conditions of industrial processing. Thermostable xylanases display high-specific activity at elevated temperatures and distinguish themselves in biochemical properties, structures, and modes of action from their mesophilic counterparts. Natural xylanases can be further improved through genetic engineering. Rapid progress with genome editing, writing, and synthetic biological techniques have provided unlimited potential to produce thermophilic xylanases in their natural hosts or cell factories including bacteria, yeasts, and filamentous fungi. This review will discuss the biotechnological potential of xylanases from thermophilic microorganisms and the ways they are being optimized and produced for various industrial applications.

  5. Thermincola carboxydiphila gen. nov., sp. nov., a novel anaerobic, carboxydotrophic, hydrogenogenic bacterium from a hot spring of the Lake Baikal area.

    PubMed

    Sokolova, Tatyana G; Kostrikina, Nadezhda A; Chernyh, Nikolai A; Kolganova, Tatjana V; Tourova, Tatjana P; Bonch-Osmolovskaya, Elizaveta A

    2005-09-01

    A novel anaerobic, thermophilic, alkalitolerant bacterium, strain 2204(T), was isolated from a hot spring of the Baikal Lake region. The cells of strain 2204(T) were straight rods of variable length, Gram-positive with an S-layer, motile with one to two lateral flagella, and often formed aggregates of 3-15 cells. The isolate was shown to be an obligate anaerobe oxidizing CO and producing equimolar quantities of H(2) and CO(2) according to the equation CO+H(2)O-->CO(2)+H(2). No organic substrates were used as energy sources. For lithotrophic growth on CO, 0.2 g acetate or yeast extract l(-1) was required but did not support growth in the absence of CO. Growth was observed in the temperature range 37-68 degrees C, the optimum being 55 degrees C. The pH range for growth was 6.7-9.5, the optimum pH being 8.0. The generation time under optimal conditions was 1.3 h. The DNA G+C content was 45 mol%. Penicillin, erythromycin, streptomycin, rifampicin, vancomycin and tetracycline completely inhibited both growth and CO utilization by strain 2204(T). Thus, isolate 2204(T) was found to be the first known moderately thermophilic and alkalitolerant H(2)-producing anaerobic carboxydotroph. The novel bacterium fell within the cluster of the family Peptococcaceae within the low-G+C-content Gram-positive bacteria, where it formed a separate branch. On the basis of morphological, physiological and phylogenetic features, strain 2204(T) should be assigned to a novel genus and species, for which the name Thermincola carboxydiphila gen. nov., sp. nov. is proposed. The type strain is strain 2204(T) (=DSM 17129(T)=VKM B-2283(T)=JCM 13258(T)).

  6. Complete genome sequence of Thermovibrio ammonificans HB-1T, a thermophilic, chemolithoautotrophic bacterium isolated from a deep-sea hydrothermal vent

    PubMed Central

    Giovannelli, Donato; Ricci, Jessica; Pérez-Rodríguez, Ileana; Hügler, Michael; O’Brien, Charles; Keddis, Ramaydalis; Grosche, Ashley; Goodwin, Lynne; Bruce, David; Davenport, Karen W.; Detter, Chris; Han, James; Han, Shunsheng; Ivanova, Natalia; Land, Miriam L.; Mikhailova, Natalia; Nolan, Matt; Pitluck, Sam; Tapia, Roxanne; Woyke, Tanja

    2012-01-01

    Thermovibrio ammonificans type strain HB-1T is a thermophilic (Topt: 75°C), strictly anaerobic, chemolithoautotrophic bacterium that was isolated from an active, high temperature deep-sea hydrothermal vent on the East Pacific Rise. This organism grows on mineral salts medium in the presence of CO2/H2, using NO3- or S0 as electron acceptors, which are reduced to ammonium or hydrogen sulfide, respectively. T. ammonificans is one of only three species within the genus Thermovibrio, a member of the family Desulfurobacteriaceae, and it forms a deep branch within the phylum Aquificae. Here we report the main features of the genome of T. ammonificans strain HB-1T (DSM 15698T). PMID:23449845

  7. Fate of pathogen indicators in a domestic blend of food waste and wastewater through a two-stage anaerobic digestion system.

    PubMed

    Rounsefell, B D; O'Sullivan, C A; Chinivasagam, N; Batstone, D; Clarke, W P

    2013-01-01

    Anaerobic digestion is a viable on-site treatment technology for rich organic waste streams such as food waste and blackwater. In contrast to large-scale municipal wastewater treatment plants which are typically located away from the community, the effluent from any type of on-site system is a potential pathogenic hazard because of the intimacy of the system to the community. The native concentrations of the pathogen indicators Escherichia coli, Clostridium perfringens and somatic coliphage were tracked for 30 days under stable operation (organic loading rate (OLR) = 1.8 kgCOD m(-3) day(-1), methane yield = 52% on a chemical oxygen demand (COD) basis) of a two-stage laboratory-scale digester treating a mixture of food waste and blackwater. E. coli numbers were reduced by a factor of 10(6.4) in the thermophilic stage, from 10(7.5±0.3) to 10(1.1±0.1) cfu 100 mL(-1), but regenerated by a factor of 10(4) in the mesophilic stage. Neither the thermophilic nor mesophilic stages had any significant impact on C. perfringens concentrations. Coliphage concentrations were reduced by a factor of 10(1.4) across the two stages. The study shows that anaerobic digestion only reduces pathogen counts marginally but that counts in effluent samples could be readily reduced to below detection limits by filtration through a 0.22 µm membrane, to investigate membrane filtration as a possible sanitation technique.

  8. Stress response physiology of thermophiles.

    PubMed

    Ranawat, Preeti; Rawat, Seema

    2017-04-01

    Thermo (or hyperthermo) philic microorganisms are ubiquitous having a wide range of habitats from freshly fallen snow to pasteurized milk to geothermal areas like hot springs. The variations in physicochemical conditions, viz., temperature, pH, nutrient availability and light intensity in the habitats always pose stress conditions for the inhabitants leading to slow growth or cell death. The industrial processes used for harvesting secondary metabolites such as enzymes, toxins and organic acids also create stressed environments for thermophiles. The production of DNA-binding proteins, activation of reactive oxygen species detoxification system, compatible solute accumulation, expression of heat shock proteins and alterations in morphology are a few examples of physiological changes demonstrated by these microscopic lifeforms in stress. These microorganisms exhibit complex genetic and physiological changes to minimize, adapt to and repair damage caused by extreme environmental disturbances. These changes are termed as 'stress responses' which enable them to stabilize their homeostasis. The exploration of important thermophilic factors would pave the way in engineering the microbial strains for various biotechnological applications. This review article presents a picture of physiological responses of thermophiles against various stress conditions as their mechanisms to respond to stress make them model organisms to further explore them for basic and applied biology purposes.

  9. Deferribacter thermophilus gen. nov., sp. nov., a novel thermophilic manganese- and iron-reducing bacterium isolated from a petroleum reservoir.

    PubMed

    Greene, A C; Patel, B K; Sheehy, A J

    1997-04-01

    A thermophilic anaerobic bacterium, designated strain BMAT (T = type strain), was isolated from the production water of Beatrice oil field in the North Sea (United Kingdom). The cells were straight to bent rods (1 to 5 by 0.3 to 0.5 microns) which stained gram negative. Strain BMAT obtained energy from the reduction of manganese (IV), iron(III), and nitrate in the presence of yeast extract, peptone, Casamino Acids, tryptone, hydrogen, malate, acetate, citrate, pyruvate, lactate, succinate, and valerate. The isolate grew optimally at 60 degrees C (temperature range for growth, 50 to 65 degrees C) and in the presence of 2% (wt/vol) NaCl (NaCl range for growth, 0 to 5% [wt/vol]). The DNA base composition was 34 mol% G + C. Phylogenetic analyses of the 16S rRNA gene indicated that strain BMAT is a member of the domain Bacteria. The closest known bacterium is the moderate thermophile Flexistipes sinusarabici (similarity value, 88%). Strain BMAT possesses phenotypic and phylogenetic traits that do not allow its classification as a member of any previously described genus; therefore, we propose that this isolate should be described as a member of a novel species of a new genus, Deferribacter thermophilus gen. nov., sp. nov.

  10. Bioprospecting thermophiles for cellulase production: a review.

    PubMed

    Acharya, Somen; Chaudhary, Anita

    2012-07-01

    Most of the potential bioprospecting is currently related to the study of the extremophiles and their potential use in industrial processes. Recently microbial cellulases find applications in various industries and constitute a major group of industrial enzymes. Considerable amount of work has been done on microbial cellulases, especially with resurgence of interest in biomass ethanol production employing cellulases and use of cellulases in textile and paper industry. Most efficient method of lignocellulosic biomass hydrolysis is through enzymatic saccharification using cellulases. Significant information has also been gained about the physiology of thermophilic cellulases producers and process development for enzyme production and biomass saccharification. The review discusses the current knowledge on cellulase producing thermophilic microorganisms, their physiological adaptations and control of cellulase gene expression. It discusses the industrial applications of thermophilic cellulases, their cost of production and challenges in cellulase research especially in the area of improving process economics of enzyme production.

  11. Relationship between microbial community dynamics and process performance during thermophilic sludge bioleaching.

    PubMed

    Chen, Shen-Yi; Chou, Li-Chieh

    2016-08-01

    Heavy metals can be removed from the sludge using bioleaching technologies at thermophilic condition, thereby providing an option for biotreatment of wasted sludge generated from wastewater treatment. The purposes of this study were to establish a molecular biology technique, real-time PCR, for the detection and enumeration of the sulfur-oxidizing bacteria during the thermophilic sludge bioleaching. The 16S rRNA gene for real-time PCR quantification targeted the bioleaching bacteria: Sulfobacillus thermosulfidooxidans, Sulfobacillus acidophilus, and Acidithiobacillus caldus. The specificity and stringency for thermophilic sulfur-oxidizing bacteria were tested before the experiments of monitoring the bacterial community, bacterial number during the thermophilic sludge bioleaching and the future application on testing various environmental samples. The results showed that S. acidophilus was identified as the dominant sulfur-oxidizing bacteria, while A. caldus and S. thermosulfidooxidans occurred in relatively low numbers. The total number of the sulfur-oxidizing bacteria increased during the thermophilic bioleaching process. Meanwhile, the decrease of pH, production of sulfate, degradation of SS/VSS, and solubilization of heavy metal were found to correlate well with the population of thermophilic sulfur-oxidizing bacteria during the bioleaching process. The real-time PCR used in this study is a suitable method to monitor numbers of thermophilic sulfur-oxidizing bacteria during the bioleaching process.

  12. Comparing Residue Clusters from Thermophilic and Mesophilic Enzymes Reveals Adaptive Mechanisms.

    PubMed

    Sammond, Deanne W; Kastelowitz, Noah; Himmel, Michael E; Yin, Hang; Crowley, Michael F; Bomble, Yannick J

    2016-01-01

    Understanding how proteins adapt to function at high temperatures is important for deciphering the energetics that dictate protein stability and folding. While multiple principles important for thermostability have been identified, we lack a unified understanding of how internal protein structural and chemical environment determine qualitative or quantitative impact of evolutionary mutations. In this work we compare equivalent clusters of spatially neighboring residues between paired thermophilic and mesophilic homologues to evaluate adaptations under the selective pressure of high temperature. We find the residue clusters in thermophilic enzymes generally display improved atomic packing compared to mesophilic enzymes, in agreement with previous research. Unlike residue clusters from mesophilic enzymes, however, thermophilic residue clusters do not have significant cavities. In addition, anchor residues found in many clusters are highly conserved with respect to atomic packing between both thermophilic and mesophilic enzymes. Thus the improvements in atomic packing observed in thermophilic homologues are not derived from these anchor residues but from neighboring positions, which may serve to expand optimized protein core regions.

  13. Bioprospecting thermophiles for cellulase production: a review

    PubMed Central

    Acharya, Somen; Chaudhary, Anita

    2012-01-01

    Most of the potential bioprospecting is currently related to the study of the extremophiles and their potential use in industrial processes. Recently microbial cellulases find applications in various industries and constitute a major group of industrial enzymes. Considerable amount of work has been done on microbial cellulases, especially with resurgence of interest in biomass ethanol production employing cellulases and use of cellulases in textile and paper industry. Most efficient method of lignocellulosic biomass hydrolysis is through enzymatic saccharification using cellulases. Significant information has also been gained about the physiology of thermophilic cellulases producers and process development for enzyme production and biomass saccharification. The review discusses the current knowledge on cellulase producing thermophilic microorganisms, their physiological adaptations and control of cellulase gene expression. It discusses the industrial applications of thermophilic cellulases, their cost of production and challenges in cellulase research especially in the area of improving process economics of enzyme production. PMID:24031898

  14. Deletion of the hfsB gene increases ethanol production in Thermoanaerobacterium saccharolyticum and several other thermophilic anaerobic bacteria

    DOE PAGES

    Eminoglu, Aysenur; Murphy, Sean Jean-Loup; Maloney, Marybeth; ...

    2017-11-30

    With the discovery of interspecies hydrogen transfer in the late 1960s (Bryant et al. in Arch Microbiol 59:20–31, 1967), it was shown that reducing the partial pressure of hydrogen could cause mixed acid fermenting organisms to produce acetate at the expense of ethanol. Hydrogen and ethanol are both more reduced than glucose. Thus there is a tradeoff between production of these compounds imposed by electron balancing requirements; however, the mechanism is not fully known. Deletion of the hfsA or B subunits resulted in a roughly 1.8-fold increase in ethanol yield. The increase in ethanol production appears to be associated withmore » an increase in alcohol dehydrogenase activity, which appears to be due, at least in part, to increased expression of the adhE gene, and may suggest a regulatory linkage between hfsB and adhE. We studied this system most intensively in the organism Thermoanaerobacterium saccharolyticum; however, deletion of hfsB also increases ethanol production in other thermophilic bacteria suggesting that this could be used as a general technique for engineering thermophilic bacteria for improved ethanol production in organisms with hfs-type hydrogenases. Since its discovery by Shaw et al. (JAMA 191:6457–64, 2009), the hfs hydrogenase has been suspected to act as a regulator due to the presence of a PAS domain. We provide additional support for the presence of a regulatory phenomenon. In addition, we find a practical application for this scientific insight, namely increasing ethanol yield in strains that are of interest for ethanol production from cellulose or hemicellulose. In two of these organisms (T. xylanolyticum and T. thermosaccharolyticum), the ethanol yields are the highest reported to date.« less

  15. Deletion of the hfsB gene increases ethanol production in Thermoanaerobacterium saccharolyticum and several other thermophilic anaerobic bacteria

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Eminoglu, Aysenur; Murphy, Sean Jean-Loup; Maloney, Marybeth

    With the discovery of interspecies hydrogen transfer in the late 1960s (Bryant et al. in Arch Microbiol 59:20–31, 1967), it was shown that reducing the partial pressure of hydrogen could cause mixed acid fermenting organisms to produce acetate at the expense of ethanol. Hydrogen and ethanol are both more reduced than glucose. Thus there is a tradeoff between production of these compounds imposed by electron balancing requirements; however, the mechanism is not fully known. Deletion of the hfsA or B subunits resulted in a roughly 1.8-fold increase in ethanol yield. The increase in ethanol production appears to be associated withmore » an increase in alcohol dehydrogenase activity, which appears to be due, at least in part, to increased expression of the adhE gene, and may suggest a regulatory linkage between hfsB and adhE. We studied this system most intensively in the organism Thermoanaerobacterium saccharolyticum; however, deletion of hfsB also increases ethanol production in other thermophilic bacteria suggesting that this could be used as a general technique for engineering thermophilic bacteria for improved ethanol production in organisms with hfs-type hydrogenases. Since its discovery by Shaw et al. (JAMA 191:6457–64, 2009), the hfs hydrogenase has been suspected to act as a regulator due to the presence of a PAS domain. We provide additional support for the presence of a regulatory phenomenon. In addition, we find a practical application for this scientific insight, namely increasing ethanol yield in strains that are of interest for ethanol production from cellulose or hemicellulose. In two of these organisms (T. xylanolyticum and T. thermosaccharolyticum), the ethanol yields are the highest reported to date.« less

  16. Biogasification of sorghum in a novel anaerobic digester

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Srivastava, V.J.; Biljetina, R.; Isaacson, H.R.

    1987-01-01

    The Institute of Gas Technology (IGT) conducted pilot-scale anaerobic digestion experiments with ensiled sorghum in a 160 ft/sup 3/ digester at the experimental test unit (ETU) facility at the Walt Disney World Resort Complex in Florida. The study focused on improving bioconversion efficiencies and process stability by employing a novel reactor concept developed at IGT. Steady-state performance data were collected from the ETU as well as from a laboratory-scale conventional stirred tank reactor (CSTR) at loading rates of 0.25 and 0.50 lb organic matter/ft/sup 3/-day at mesophilic and thermophilic temperatures, respectively. This paper will describe the ETU facility, novel digestermore » design and operating techniques, and the results obtained during 12 months of stable and uninterrupted operation of the ETU and the CSTR which showed that methane yields anad rates from the ETU were 20% to 50% higher than those of the CSTR. 10 refs., 7 figs., 5 tabs.« less

  17. Probing the Folding-Unfolding Transition of a Thermophilic Protein, MTH1880

    PubMed Central

    Jung, Youngjin; Han, Jeongmin; Yun, Ji-Hye; Chang, Iksoo; Lee, Weontae

    2016-01-01

    The folding mechanism of typical proteins has been studied widely, while our understanding of the origin of the high stability of thermophilic proteins is still elusive. Of particular interest is how an atypical thermophilic protein with a novel fold maintains its structure and stability under extreme conditions. Folding-unfolding transitions of MTH1880, a thermophilic protein from Methanobacterium thermoautotrophicum, induced by heat, urea, and GdnHCl, were investigated using spectroscopic techniques including circular dichorism, fluorescence, NMR combined with molecular dynamics (MD) simulations. Our results suggest that MTH1880 undergoes a two-state N to D transition and it is extremely stable against temperature and denaturants. The reversibility of refolding was confirmed by spectroscopic methods and size exclusion chromatography. We found that the hyper-stability of the thermophilic MTH1880 protein originates from an extensive network of both electrostatic and hydrophobic interactions coordinated by the central β-sheet. Spectroscopic measurements, in combination with computational simulations, have helped to clarify the thermodynamic and structural basis for hyper-stability of the novel thermophilic protein MTH1880. PMID:26766214

  18. Comparing residue clusters from thermophilic and mesophilic enzymes reveals adaptive mechanisms

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Sammond, Deanne W.; Kastelowitz, Noah; Himmel, Michael E.

    Understanding how proteins adapt to function at high temperatures is important for deciphering the energetics that dictate protein stability and folding. While multiple principles important for thermostability have been identified, we lack a unified understanding of how internal protein structural and chemical environment determine qualitative or quantitative impact of evolutionary mutations. In this work we compare equivalent clusters of spatially neighboring residues between paired thermophilic and mesophilic homologues to evaluate adaptations under the selective pressure of high temperature. We find the residue clusters in thermophilic enzymes generally display improved atomic packing compared to mesophilic enzymes, in agreement with previous research.more » Unlike residue clusters from mesophilic enzymes, however, thermophilic residue clusters do not have significant cavities. In addition, anchor residues found in many clusters are highly conserved with respect to atomic packing between both thermophilic and mesophilic enzymes. As a result, the improvements in atomic packing observed in thermophilic homologues are not derived from these anchor residues but from neighboring positions, which may serve to expand optimized protein core regions.« less

  19. Comparing residue clusters from thermophilic and mesophilic enzymes reveals adaptive mechanisms

    DOE PAGES

    Sammond, Deanne W.; Kastelowitz, Noah; Himmel, Michael E.; ...

    2016-01-07

    Understanding how proteins adapt to function at high temperatures is important for deciphering the energetics that dictate protein stability and folding. While multiple principles important for thermostability have been identified, we lack a unified understanding of how internal protein structural and chemical environment determine qualitative or quantitative impact of evolutionary mutations. In this work we compare equivalent clusters of spatially neighboring residues between paired thermophilic and mesophilic homologues to evaluate adaptations under the selective pressure of high temperature. We find the residue clusters in thermophilic enzymes generally display improved atomic packing compared to mesophilic enzymes, in agreement with previous research.more » Unlike residue clusters from mesophilic enzymes, however, thermophilic residue clusters do not have significant cavities. In addition, anchor residues found in many clusters are highly conserved with respect to atomic packing between both thermophilic and mesophilic enzymes. As a result, the improvements in atomic packing observed in thermophilic homologues are not derived from these anchor residues but from neighboring positions, which may serve to expand optimized protein core regions.« less

  20. Genetic tool development underpins recent advances in thermophilic whole‐cell biocatalysts

    PubMed Central

    Taylor, M. P.; van Zyl, L.; Tuffin, I. M.; Leak, D. J.; Cowan, D. A.

    2011-01-01

    Summary The environmental value of sustainably producing bioproducts from biomass is now widely appreciated, with a primary target being the economic production of fuels such as bioethanol from lignocellulose. The application of thermophilic prokaryotes is a rapidly developing niche in this field, driven by their known catabolic versatility with lignocellulose‐derived carbohydrates. Fundamental to the success of this work has been the development of reliable genetic and molecular systems. These technical tools are now available to assist in the development of other (hyper)thermophilic strains with diverse phenotypes such as hemicellulolytic and cellulolytic properties, branched chain alcohol production and other ‘valuable bioproduct’ synthetic capabilities. Here we present an insight into the historical limitations, recent developments and current status of a number of genetic systems for thermophiles. We also highlight the value of reliable genetic methods for increasing our knowledge of thermophile physiology. We argue that the development of robust genetic systems is paramount in the evolution of future thermophilic based bioprocesses and make suggestions for future approaches and genetic targets that will facilitate this process. PMID:21310009

  1. Comparing Residue Clusters from Thermophilic and Mesophilic Enzymes Reveals Adaptive Mechanisms

    PubMed Central

    Sammond, Deanne W.; Kastelowitz, Noah; Himmel, Michael E.; Yin, Hang; Crowley, Michael F.; Bomble, Yannick J.

    2016-01-01

    Understanding how proteins adapt to function at high temperatures is important for deciphering the energetics that dictate protein stability and folding. While multiple principles important for thermostability have been identified, we lack a unified understanding of how internal protein structural and chemical environment determine qualitative or quantitative impact of evolutionary mutations. In this work we compare equivalent clusters of spatially neighboring residues between paired thermophilic and mesophilic homologues to evaluate adaptations under the selective pressure of high temperature. We find the residue clusters in thermophilic enzymes generally display improved atomic packing compared to mesophilic enzymes, in agreement with previous research. Unlike residue clusters from mesophilic enzymes, however, thermophilic residue clusters do not have significant cavities. In addition, anchor residues found in many clusters are highly conserved with respect to atomic packing between both thermophilic and mesophilic enzymes. Thus the improvements in atomic packing observed in thermophilic homologues are not derived from these anchor residues but from neighboring positions, which may serve to expand optimized protein core regions. PMID:26741367

  2. Anoxybacter fermentans gen. nov., sp. nov., a piezophilic, thermophilic, anaerobic, fermentative bacterium isolated from a deep-sea hydrothermal vent.

    PubMed

    Zeng, Xiang; Zhang, Zhao; Li, Xi; Zhang, Xiaobo; Cao, Junwei; Jebbar, Mohamed; Alain, Karine; Shao, Zongze

    2015-02-01

    A novel piezophilic, thermophilic, anaerobic, fermentative bacterial strain, designated strain DY22613(T), was isolated from a deep-sea hydrothermal sulfide deposit at the East Pacific Rise (GPS position: 102.6° W 3.1° S). Cells of strain DY22613(T) were long, motile rods (10 to 20 µm in length and 0.5 µm in width) with peritrichous flagella and were Gram-stain-negative. Growth was recorded at 44-72 °C (optimum 60-62 °C) and at hydrostatic pressures of 0.1-55 MPa (optimum 20 MPa). The pH range for growth was from pH 5.0 to 9.0 with an optimum at pH 7.0. Growth was observed in the presence of 1 to 8 % (w/v) sea salts and 0.65 to 5.2 % (w/v) NaCl, with optimum salt concentrations at 3.5 % for sea salts and at 2.3 % for NaCl. Under optimal growth conditions, the shortest generation time observed was 27 min (60 °C, 20 MPa). Strain DY22613(T) was heterotrophic, able to utilize complex organic compounds, amino acids, sugars and organic acids including peptone, tryptone, beef extract, yeast extract, alanine, glutamine, methionine, phenylalanine, serine, threonine, fructose, fucose, galactose, gentiobiose, glucose, mannose, melibiose, palatinose, rhamnose, turanose, pyruvate, lactic acid, methyl ester, erythritol, galacturonic acid and glucosaminic acid. Strain DY22613(T) was able to reduce Fe(III) compounds, including Fe(III) oxyhydroxide (pH 7.0), amorphous iron(III) oxide (pH 9.0), goethite (α-FeOOH, pH 12.0), Fe(III) citrate and elementary sulfur. Products of fermentation were butyrate, acetate and hydrogen. Main cellular fatty acids were iso-C15 : 0, iso-C14 : 0 3-OH and C14 : 0. The genomic DNA G+C content of strain DY22613(T) was 36.7 mol%. Based on 16S rRNA gene sequence analysis, the strain forms a novel lineage within the class Clostridia and clusters with the order Haloanaerobiales (86.92 % 16S rRNA gene sequence similarity). The phylogenetic data suggest that the lineage represents at least a novel genus and species, for which the name Anoxybacter

  3. Treatment of synthetic kraft evaporator condensate using thermophilic and mesophilic membrane aerated biofilm reactors.

    PubMed

    Liao, B Q; Zheng, M R; Ratana-Rueangsri, L

    2010-01-01

    A comparative study on the treatment of synthetic kraft evaporator condensate was conducted using thermophilic (55 degrees C) and mesophilic (30 degrees C) membrane aerated biofilm reactors (MABRs) and sequencing batch reactors (SBRs) for 8 months. Under tested conditions, a chemical oxygen demand (COD) removal efficiency of 80-95% was achieved with both thermophilic and mesophilic MABRs and SBRs. The COD removal efficiency of thermophilic MABR (80-90%) was slightly lower than that of the mesophilic MABR (85-95%) and the thermophilic SBR (90-95%). A significant amount (13-37%) of COD was stripped by conventional aeration in the SBRs, while stripping in MABRs was negligible. Simultaneous COD removal and denitrification were observed in the mesophilic MABR, while the thermophilic MABR contributed mainly for COD removal. Nitrification was not significant in both the thermophilic and mesophilic MABRs. The results suggest that treatment of kraft evaporator condensate is feasible with the use of both thermophilic and mesophilic MABRs in terms of COD removal with the advantages of negligible stripping.

  4. Thermophilic growth and enzymatic thermostability are polyphyletic traits within Chaetomiaceae.

    PubMed

    van den Brink, Joost; Facun, Kryss; de Vries, Michel; Stielow, J Benjamin

    2015-12-01

    Thermophilic fungi have the potential to produce industrial-relevant thermostable enzymes, in particular for the degradation of plant biomass. Sordariales is one of the few fungal orders containing several thermophilic taxa, of which many have been associated with the production of thermostable enzymes. The evolutionary affiliation of Sordariales fungi, especially between thermophiles and non-thermophilic relatives, is however poorly understood. Phylogenetic analysis within the current study was based on sequence data, derived from a traditional Sanger and highly multiplexed targeted next generation sequencing approach of 45 isolates. The inferred phylogeny and detailed growth analysis rendered the trait 'thermophily' as polyphyletic within Chaetomiaceae (Sordariales, Sordariomycetes), and characteristic to: Myceliophthora spp., Thielavia terrestris, Chaetomium thermophilum, and Mycothermus thermophilus. Compared to mesophiles, the isolates within thermophilic taxa produced enzyme mixtures with the highest thermostability of known cellulase activities. Temperature profiles of the enzyme activities correlated strongly with the optimal growth temperatures of the isolates but not with their phylogenetic relationships. This strong correlation between growth and enzyme characteristics indicated that detailed analysis of growth does give predictive information on enzyme physiology. The variation in growth and enzyme characteristics reveals these fungi as an excellent platform to better understand fungal thermophily and enzyme thermostability. Copyright © 2015 The British Mycological Society. Published by Elsevier Ltd. All rights reserved.

  5. Glycoside hydrolase activities of thermophilic bacterial consortia adapted to switchgrass.

    PubMed

    Gladden, John M; Allgaier, Martin; Miller, Christopher S; Hazen, Terry C; VanderGheynst, Jean S; Hugenholtz, Philip; Simmons, Blake A; Singer, Steven W

    2011-08-15

    Industrial-scale biofuel production requires robust enzymatic cocktails to produce fermentable sugars from lignocellulosic biomass. Thermophilic bacterial consortia are a potential source of cellulases and hemicellulases adapted to harsher reaction conditions than commercial fungal enzymes. Compost-derived microbial consortia were adapted to switchgrass at 60°C to develop thermophilic biomass-degrading consortia for detailed studies. Microbial community analysis using small-subunit rRNA gene amplicon pyrosequencing and short-read metagenomic sequencing demonstrated that thermophilic adaptation to switchgrass resulted in low-diversity bacterial consortia with a high abundance of bacteria related to thermophilic paenibacilli, Rhodothermus marinus, and Thermus thermophilus. At lower abundance, thermophilic Chloroflexi and an uncultivated lineage of the Gemmatimonadetes phylum were observed. Supernatants isolated from these consortia had high levels of xylanase and endoglucanase activities. Compared to commercial enzyme preparations, the endoglucanase enzymes had a higher thermotolerance and were more stable in the presence of 1-ethyl-3-methylimidazolium acetate ([C2mim][OAc]), an ionic liquid used for biomass pretreatment. The supernatants were used to saccharify [C2mim][OAc]-pretreated switchgrass at elevated temperatures (up to 80°C), demonstrating that these consortia are an excellent source of enzymes for the development of enzymatic cocktails tailored to more extreme reaction conditions.

  6. Genomic Analysis of Caldithrix abyssi, the Thermophilic Anaerobic Bacterium of the Novel Bacterial Phylum Calditrichaeota

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Kublanov, Ilya V.; Sigalova, Olga M.; Gavrilov, Sergey N.

    The genome of Caldithrix abyssi, the first cultivated representative of a phylum-level bacterial lineage, was sequenced within the framework of Genomic Encyclopedia of Bacteria and Archaea (GEBA) project. The genomic analysis revealed mechanisms allowing this anaerobic bacterium to ferment peptides or to implement nitrate reduction with acetate or molecular hydrogen as electron donors. The genome encoded five different [NiFe]- and [FeFe]-hydrogenases, one of which, group 1 [NiFe]-hydrogenase, is presumably involved in lithoheterotrophic growth, three other produce H 2 during fermentation, and one is apparently bidirectional. The ability to reduce nitrate is determined by a nitrate reductase of the Nap family,more » while nitrite reduction to ammonia is presumably catalyzed by an octaheme cytochrome c nitrite reductase εHao. The genome contained genes of respiratory polysulfide/thiosulfate reductase, however, elemental sulfur and thiosulfate were not used as the electron acceptors for anaerobic respiration with acetate or H 2, probably due to the lack of the gene of the maturation protein. Nevertheless, elemental sulfur and thiosulfate stimulated growth on fermentable substrates (peptides), being reduced to sulfide, most probably through the action of the cytoplasmic sulfide dehydrogenase and/or NAD(P)-dependent [NiFe]-hydrogenase (sulfhydrogenase) encoded by the genome. Surprisingly, the genome of this anaerobic microorganism encoded all genes for cytochrome c oxidase, however, its maturation machinery seems to be non-operational due to genomic rearrangements of supplementary genes. Despite the fact that sugars were not among the substrates reported when C. abyssi was first described, our genomic analysis revealed multiple genes of glycoside hydrolases, and some of them were predicted to be secreted. This finding aided in bringing out four carbohydrates that supported the growth of C. abyssi: starch, cellobiose, glucomannan and xyloglucan. The genomic analysis

  7. Genomic Analysis of Caldithrix abyssi, the Thermophilic Anaerobic Bacterium of the Novel Bacterial Phylum Calditrichaeota

    DOE PAGES

    Kublanov, Ilya V.; Sigalova, Olga M.; Gavrilov, Sergey N.; ...

    2017-02-20

    The genome of Caldithrix abyssi, the first cultivated representative of a phylum-level bacterial lineage, was sequenced within the framework of Genomic Encyclopedia of Bacteria and Archaea (GEBA) project. The genomic analysis revealed mechanisms allowing this anaerobic bacterium to ferment peptides or to implement nitrate reduction with acetate or molecular hydrogen as electron donors. The genome encoded five different [NiFe]- and [FeFe]-hydrogenases, one of which, group 1 [NiFe]-hydrogenase, is presumably involved in lithoheterotrophic growth, three other produce H 2 during fermentation, and one is apparently bidirectional. The ability to reduce nitrate is determined by a nitrate reductase of the Nap family,more » while nitrite reduction to ammonia is presumably catalyzed by an octaheme cytochrome c nitrite reductase εHao. The genome contained genes of respiratory polysulfide/thiosulfate reductase, however, elemental sulfur and thiosulfate were not used as the electron acceptors for anaerobic respiration with acetate or H 2, probably due to the lack of the gene of the maturation protein. Nevertheless, elemental sulfur and thiosulfate stimulated growth on fermentable substrates (peptides), being reduced to sulfide, most probably through the action of the cytoplasmic sulfide dehydrogenase and/or NAD(P)-dependent [NiFe]-hydrogenase (sulfhydrogenase) encoded by the genome. Surprisingly, the genome of this anaerobic microorganism encoded all genes for cytochrome c oxidase, however, its maturation machinery seems to be non-operational due to genomic rearrangements of supplementary genes. Despite the fact that sugars were not among the substrates reported when C. abyssi was first described, our genomic analysis revealed multiple genes of glycoside hydrolases, and some of them were predicted to be secreted. This finding aided in bringing out four carbohydrates that supported the growth of C. abyssi: starch, cellobiose, glucomannan and xyloglucan. The genomic analysis

  8. Screening of Thermophilic Bacteria Produce Xylanase from Sapan Sungai Aro Hot Spring South Solok

    NASA Astrophysics Data System (ADS)

    Irdawati, I.; Syamsuardi, S.; Agustien, A.; Rilda, Y.

    2018-04-01

    xylanase is one of the enzymes with great prospects as hemicellulose hydrolyzing enzyme. Global annual market demand for this enzyme reach US 200 million. This enzyme catalyzes the xylan (hemicellulose) reactions breaking into xilooligosakarida and xylose. Xylanase can be applied to various industrial sectors such as bread, sugar xylose, biofuels, especially in bleaching paper (bleaching) pulp. Xylanase Isable to replace conventional chemical bleaching using chlorine that is not friendly for the environment. Currently xylanase production is extracted from the thermophilic bacteria for enzyme stability at high temperatures that are suitable for industrial applications. Thermophilic bacteria can be isolated from a hot spring, one of the which is a source of Sapan Sungai Aro Hot Spring, located in the district South Solok. The aim of this study was to select and identification of thermophilic bacteria can produce xylanase.This roomates is a descriptive study, which was Carried out in the Laboratory of Microbiology, Mathematic and Science Faculty of Padang State University, and Laboratory of Bacteriology, BasoVeterinary Research Center. The research procedure consisted of the preparation and sterilization of materials and tools, medium manufacturing, regeneration, selection and identification. Selection is performed by using a semiquantitative screening plate that contains xylan substrate. Identification is based on microscopic and biochemical characteristics until the genus level.Selection results Showed 12 out of 16 isolates had xilanolitik activity, with the highest activity is SSA2 with xilanolitik index of 0.74. The top five index producehigestxilanolitik isolates that are SSA2, SSA3 and SSA4 identified as Bacillus sp. 1., and SSAS6 and SSA7 is Bacillus sp. 2.

  9. Comparison of laboratory-scale thermophilic biofilm and activated sludge processes integrated with a mesophilic activated sludge process.

    PubMed

    Suvilampi, J; Lehtomäki, A; Rintala, J

    2003-07-01

    A combined thermophilic-mesophilic wastewater treatment was studied using a laboratory-scale thermophilic activated sludge process (ASP) followed by mesophilic ASP or a thermophilic suspended carrier biofilm process (SCBP) followed by mesophilic ASP, both systems treating diluted molasses (dilution factor 1:500 corresponding GF/A-filtered COD (COD(filt)) of 1900+/-190 mgl(-1)). With hydraulic retention times (HRTs) of 12-18 h the thermophilic ASP and thermophilic SCBP removed 60+/-13% and 62+/-7% of COD(filt), respectively, with HRT of 8 h the removals were 48+/-1% and 69+/-4%. The sludge volume index (SVI) was notably lower in the thermophilic SCBP (measured from suspended sludge) than in the thermophilic ASP. Under the lowest HRT the mesophilic ASP gave better performance (as SVI, COD(filt), and COD(tot) removals) after the thermophilic SCBP than after the thermophilic ASP. Measured sludge yields were low (less than 0.1 kg suspended solids (SS) kg COD(filt removed)(-1)) in all processes. Both thermophilic treatments removed 80-85% of soluble COD (COD(sol)) whereas suspended COD (COD(susp)) and colloidal COD (COD(col)) were increased. Both mesophilic post-treatments removed all COD(col) and most of the COD(susp) from the thermophilic effluents. In conclusion, combined thermophilic-mesophilic treatment appeared to be easily operable and produced high effluent quality.

  10. Application of Biofilm Covered Activated Carbon Particles as a Microbial Inoculum Delivery System for Enhanced Bioaugmentation of PCBs in Contaminated Sediment

    DTIC Science & Technology

    2013-09-01

    after anaerobic digestion at thermophilic conditions (60- 70C). Application of biofilm covered activated carbon particles as a microbial inoculum...Sludge Thickener; Sludge = Sludge after anaerobic digestion at thermophilic conditions (60- 70C). C3. Microscopic evaluation of dechlorinating...associated enzymes are capable of opening the biphenyl ring structure and transform the molecule into a linear structure, this changed structure was not

  11. Thermophilic Beta-Glycosidase

    NASA Technical Reports Server (NTRS)

    Grogan, Dennis W.

    1992-01-01

    Report describes identification of thermophilic Beta-glycosidase enzyme from isolate of Sulfolobus solfataricus, sulfur-metabolizing archaebacteria growing aerobically and heterotrophically to relatively high cell yields. Enzyme useful in enzymatic conversion of cellulose to D-glucose and important in recycling of biomass. Used for removal of lactose from milk products. Offers promise as model substance for elucidation of basic principles of structural stabilization of proteins.

  12. Versatile transformations of hydrocarbons in anaerobic bacteria: substrate ranges and regio- and stereo-chemistry of activation reactions†

    PubMed Central

    Jarling, René; Kühner, Simon; Basílio Janke, Eline; Gruner, Andrea; Drozdowska, Marta; Golding, Bernard T.; Rabus, Ralf; Wilkes, Heinz

    2015-01-01

    Anaerobic metabolism of hydrocarbons proceeds either via addition to fumarate or by hydroxylation in various microorganisms, e.g., sulfate-reducing or denitrifying bacteria, which are specialized in utilizing n-alkanes or alkylbenzenes as growth substrates. General pathways for carbon assimilation and energy gain have been elucidated for a limited number of possible substrates. In this work the metabolic activity of 11 bacterial strains during anaerobic growth with crude oil was investigated and compared with the metabolite patterns appearing during anaerobic growth with more than 40 different hydrocarbons supplied as binary mixtures. We show that the range of co-metabolically formed alkyl- and arylalkyl-succinates is much broader in n-alkane than in alkylbenzene utilizers. The structures and stereochemistry of these products are resolved. Furthermore, we demonstrate that anaerobic hydroxylation of alkylbenzenes does not only occur in denitrifiers but also in sulfate reducers. We propose that these processes play a role in detoxification under conditions of solvent stress. The thermophilic sulfate-reducing strain TD3 is shown to produce n-alkylsuccinates, which are suggested not to derive from terminal activation of n-alkanes, but rather to represent intermediates of a metabolic pathway short-cutting fumarate regeneration by reverse action of succinate synthase. The outcomes of this study provide a basis for geochemically tracing such processes in natural habitats and contribute to an improved understanding of microbial activity in hydrocarbon-rich anoxic environments. PMID:26441848

  13. Mesophilic and thermophilic biotreatment of BTEX-polluted air in reactors.

    PubMed

    Mohammad, Balsam T; Veiga, María C; Kennes, Christian

    2007-08-15

    This study compares the removal of a mixture of benzene, toluene, ethylbenzene, and all three xylene isomers (BTEX) in mesophilic and thermophilic (50 degrees C) bioreactors. In the mesophilic reactor fungi became dominant after long-term operation, while bacteria dominated in the thermophilic unit. Microbial acclimation was achieved by exposing the biofilters to initial BTEX loads of 2-15 g m(-3) h(-1), at an empty bed residence time of 96 s. After adaptation, the elimination capacities ranged from 3 to 188 g m(-3) h(-1), depending on the inlet load, for the mesophilic biofilter with removal efficiencies reaching 96%. On the other hand, in the thermophilic reactor the average removal efficiency was 83% with a maximum elimination capacity of 218 g m(-3) h(-1). There was a clear positive relationship between temperature gradients as well as CO(2) production and elimination capacities across the biofilters. The gas phase was sampled at different depths along the reactors observing that the percentage pollutant removal in each section was strongly dependant on the load applied. The fate of individual alkylbenzene compounds was checked, showing the unusually high biodegradation rate of benzene at high loads under thermophilic conditions (100%) compared to its very low removal in the mesophilic reactor at such load (<10%). Such difference was less pronounced for the other pollutants. After 210 days of operation, the dry biomass content for the mesophilic and thermophilic reactors were 0.300 and 0.114 g g(-1) (support), respectively, reaching higher removals under thermophilic conditions with a lower biomass accumulation, that is, lower pressure drop. (c) 2007 Wiley Periodicals, Inc.

  14. Genetic tool development underpins recent advances in thermophilic whole-cell biocatalysts.

    PubMed

    Taylor, M P; van Zyl, L; Tuffin, I M; Leak, D J; Cowan, D A

    2011-07-01

    The environmental value of sustainably producing bioproducts from biomass is now widely appreciated, with a primary target being the economic production of fuels such as bioethanol from lignocellulose. The application of thermophilic prokaryotes is a rapidly developing niche in this field, driven by their known catabolic versatility with lignocellulose-derived carbohydrates. Fundamental to the success of this work has been the development of reliable genetic and molecular systems. These technical tools are now available to assist in the development of other (hyper)thermophilic strains with diverse phenotypes such as hemicellulolytic and cellulolytic properties, branched chain alcohol production and other 'valuable bioproduct' synthetic capabilities. Here we present an insight into the historical limitations, recent developments and current status of a number of genetic systems for thermophiles. We also highlight the value of reliable genetic methods for increasing our knowledge of thermophile physiology. We argue that the development of robust genetic systems is paramount in the evolution of future thermophilic based bioprocesses and make suggestions for future approaches and genetic targets that will facilitate this process. © 2011 The Authors. Journal compilation © 2011 Society for Applied Microbiology and Blackwell Publishing Ltd.

  15. Force-dependent melting of supercoiled DNA at thermophilic temperatures.

    PubMed

    Galburt, E A; Tomko, E J; Stump, W T; Ruiz Manzano, A

    2014-01-01

    Local DNA opening plays an important role in DNA metabolism as the double-helix must be melted before the information contained within may be accessed. Cells finely tune the torsional state of their genomes to strike a balance between stability and accessibility. For example, while mesophilic life forms maintain negatively superhelical genomes, thermophilic life forms use unique mechanisms to maintain relaxed or even positively supercoiled genomes. Here, we use a single-molecule magnetic tweezers approach to quantify the force-dependent equilibrium between DNA melting and supercoiling at high temperatures populated by Thermophiles. We show that negatively supercoiled DNA denatures at 0.5 pN lower tension at thermophilic vs. mesophilic temperatures. This work demonstrates the ability to monitor DNA supercoiling at high temperature and opens the possibility to perform magnetic tweezers assays on thermophilic systems. The data allow for an estimation of the relative energies of base-pairing and DNA bending as a function of temperature and support speculation as to different general mechanisms of DNA opening in different environments. Lastly, our results imply that average in vivo DNA tensions range between 0.3 and 1.1 pN. Copyright © 2014 Elsevier B.V. All rights reserved.

  16. Interface Matters: The Stiffness Route to Stability of a Thermophilic Tetrameric Malate Dehydrogenase

    PubMed Central

    Kalimeri, Maria; Girard, Eric; Madern, Dominique; Sterpone, Fabio

    2014-01-01

    In this work we investigate by computational means the behavior of two orthologous bacterial proteins, a mesophilic and a thermophilic tetrameric malate dehydrogenase (MalDH), at different temperatures. Namely, we quantify how protein mechanical rigidity at different length- and time-scales correlates to protein thermophilicity as commonly believed. In particular by using a clustering analysis strategy to explore the conformational space of the folded proteins, we show that at ambient conditions and at the molecular length-scale the thermophilic variant is indeed more rigid that the mesophilic one. This rigidification is the result of more efficient inter-domain interactions, the strength of which is further quantified via ad hoc free energy calculations. When considered isolated, the thermophilic domain is indeed more flexible than the respective mesophilic one. Upon oligomerization, the induced stiffening of the thermophilic protein propagates from the interface to the active site where the loop, controlling the access to the catalytic pocket, anchors down via an extended network of ion-pairs. On the contrary in the mesophilic tetramer the loop is highly mobile. Simulations at high temperature, could not re-activate the mobility of the loop in the thermophile. This finding opens questions on the similarities of the binding processes for these two homologues at their optimal working temperature and suggests for the thermophilic variant a possible cooperative role of cofactor/substrate. PMID:25437494

  17. Interface matters: the stiffness route to stability of a thermophilic tetrameric malate dehydrogenase.

    PubMed

    Kalimeri, Maria; Girard, Eric; Madern, Dominique; Sterpone, Fabio

    2014-01-01

    In this work we investigate by computational means the behavior of two orthologous bacterial proteins, a mesophilic and a thermophilic tetrameric malate dehydrogenase (MalDH), at different temperatures. Namely, we quantify how protein mechanical rigidity at different length- and time-scales correlates to protein thermophilicity as commonly believed. In particular by using a clustering analysis strategy to explore the conformational space of the folded proteins, we show that at ambient conditions and at the molecular length-scale the thermophilic variant is indeed more rigid that the mesophilic one. This rigidification is the result of more efficient inter-domain interactions, the strength of which is further quantified via ad hoc free energy calculations. When considered isolated, the thermophilic domain is indeed more flexible than the respective mesophilic one. Upon oligomerization, the induced stiffening of the thermophilic protein propagates from the interface to the active site where the loop, controlling the access to the catalytic pocket, anchors down via an extended network of ion-pairs. On the contrary in the mesophilic tetramer the loop is highly mobile. Simulations at high temperature, could not re-activate the mobility of the loop in the thermophile. This finding opens questions on the similarities of the binding processes for these two homologues at their optimal working temperature and suggests for the thermophilic variant a possible cooperative role of cofactor/substrate.

  18. Investigation of Poultry Waste for Anaerobic Digestion: A Case Study

    NASA Astrophysics Data System (ADS)

    Salam, Christopher R.

    Anaerobic Digestion (AD) is a biological conversion technology which is being used to produce bioenergy all over the world. This energy is created from biological feedstocks, and can often use waste products from various food and agricultural processors. Biogas from AD can be used as a fuel for heating or for co-generation of electricity and heat and is a renewable substitute to using fossil fuels. Nutrient recycling and waste reduction are additional benefits, creating a final product that can be used as a fertilizer in addition to energy benefits. This project was conducted to investigate the viability of three turkey production wastes as AD feedstock: two turkey litters and a material separated from the turkey processing wastewater using dissolved air flotation (DAF) process. The DAF waste contained greases, oils and other non-commodity portions of the turkey. Using a variety of different process methods, types of bacteria, loading rates and food-to-microorganism ratios, optimal loading rates for the digestion of these three materials were obtained. In addition, the co-digestion of these materials revealed additional energy benefits. In this study, batch digestion tests were carried out to treat these three feedstocks, using mesophilic and thermophilic bacteria, using loading rates of 3 and 6 gVS/L They were tested separately and also as a mixture for co-digestion. The batch reactor used in this study had total and working volumes of 1130 mL and 500 mL, respectively. The initial organic loading was set to be 3 gVS/L, and the food to microorganism ratio was either 0.6 or 1.0 for different treatments based on the characteristics of each material. Only thermophilic (50 +/- 2ºC) temperatures were tested for the litter and DAF wastes in continuous digestion, but mesophilic and thermophilic batch digestion experiments were conducted. The optimum digestion time for all experiments was 14 days. The biogas yields of top litter, mixed litter, and DAF waste under

  19. Behaviour of emerging contaminants in sewage sludge after anaerobic digestion.

    PubMed

    Boix, C; Ibáñez, M; Fabregat-Safont, D; Morales, E; Pastor, L; Sancho, J V; Sánchez-Ramírez, J E; Hernández, F

    2016-11-01

    Nowadays, there is an increasing concern over the presence of contaminants in the aquatic environment, where they can be introduced from wastewater after their incomplete removal in the treatment plants. In this work, degradation of selected emerging pollutants in the aqueous and solid phases of sewage sludge has been investigated after anaerobic digestion using two different digesters: mesophilic and thermophilic. Initially, sludge samples were screened by ultra-high-performance liquid chromatography coupled to quadrupole time-of-flight mass spectrometry (UHPLC-QTOF MS) for identification of emerging contaminants in the samples. In a second step, a target quantitative method based on LC coupled to tandem MS was applied for selected pollutants identified in the previous screening. The behaviour of the compounds under anaerobic conditions was studied estimating the degradation efficiency and distribution of compounds between both sludge phases. Irbesartan and benzoylecgonine seemed to be notably degraded in both phases of the sludge. Venlafaxine showed a significant concentration decrease in the aqueous phase in parallel to an increase in the solid phase. The majority of the compounds showed an increase of their concentrations in both phases after the digestion. Concentrations in the solid phase were commonly higher than in the aqueous for most contaminants, indicating that they were preferentially adsorbed onto the solid particles. Copyright © 2016 Elsevier Ltd. All rights reserved.

  20. Isolation from soil and properties of the extreme thermophile Clostridium thermohydrosulfuricum.

    PubMed Central

    Wiegel, J; Ljungdahl, L G; Rawson, J R

    1979-01-01

    Thirteen strains of a strict anaerobic, extreme thermophilic bacterium were isolated from soil samples of moderate temperature, from a sewage plant in Georgia, and from hot springs in Utah and Wyoming. They were identified as strains of Clostridium thermohydrosulfuricum. The guanosine + cytosine content (moles percent) was 37.6 (determined by buoyant density) and 34.1 (determined by melting temperature). All strains required a factor present in yeast extract or tryptone growth. Growth characteristics were as follows: a pH range of 5 to 9, with the optimum between 6.9 to 7.5, in a temperature range of 40 to 78 degrees C, with the optimum at 68 degrees C. The doubling time, when grown on glucose at temperature and pH optima, was 1.2 h. The main products of glucose fermentation were ethanol, lactate, acetate, CO2, and H2. The fermentation was inhibited by H2. Formation of spores occurred easily on glucose-agar medium or when cultures growing at temperatures above 65 degrees C were allowed to cool to temperature below 55 degrees C. C. thermohydrosulfuricum occurs widely distributed in the natural environment. PMID:39062

  1. Genomic and proteomic characterization of a thermophilic Geobacillus bacteriophage GBSV1.

    PubMed

    Liu, Bin; Zhou, Fengfeng; Wu, Suijie; Xu, Ying; Zhang, Xiaobo

    2009-03-01

    Phages are present wherever life is found, and play roles in many biogeochemical and ecological processes. The thermophilic bacteriophages, however, have not been well studied. In this study, phage GBSV1 was obtained from a thermophilic bacterium Geobacillus sp. 6k51 isolated from a hot spring. GBSV1 contains a double-stranded linear DNA of 34,683bp, which encodes 54 putative open reading frames (ORFs). Thirty three of these 54 ORFs exhibit sequence similarities to genes from 7 species of Geobacillus or Bacillus bacteria, as well as of bacteriophages infecting these bacteria. Twenty-two ORFs have been functionally annotated based on both their sequence similarities to known genes and predicted Pfam protein domains. Five structural proteins of the purified GBSV1 virion have been identified by proteomic analyses. Surprisingly, 7 of the GBSV1 ORFs share sequence similarities with genes from bacteria relevant to human diseases. This is the first report that genes of human disease-inducing bacteria are found in a thermophilic phage. It is suggested that thermophilic phages may be the potential evolutionary link between thermophiles and human pathogens. The characterization of GBSV1 may possibly lead to new insights into virus-host interactions and to a better understanding of gene transfers and evolution of life on earth in general.

  2. [Treatment of Flue Gas from Sludge Drying Process by A Thermophilic Biofilter].

    PubMed

    Chen, Wen-he; Deng, Ming-jia; Luo, Hui; Ding, Wen-iie; Li, Lin; Lin, Jian; Liu, Jun-xin

    2016-01-15

    A thermophilic biofilter was employed to treat the flue gas generated from sludge drying process, and the performance in both the start period and the stationary phase was studied under the gas flow rate of 2 700-3 100 m3 x h(-1) and retention time of 21.88-25.10 s. The results showed that the thermophilic biofilter could effectively treat gases containing sulfur dioxide, ammonia and volatile organic compounds (VOC). The removal efficiencies could reach 100%, 93.61% and 87.01%, respectively. Microbial analysis indicated that most of the population belonged to thermophilic bacteria. Paenibacillus sp., Chelatococcus sp., Bacillus sp., Clostridium thermosuccinogenes, Pseudoxanthomonas sp. and Geobacillus debilis which were abundant in the thermophilic biofilter, had the abilities of denitrification, desulfurization and degradation of volatile organic compounds.

  3. Anaerobic

    MedlinePlus

    ... oxygen." The term has many uses in medicine. Anaerobic bacteria are germs that can survive and grow where ... Infections like tetanus and gangrene are caused by anaerobic bacteria. Anaerobic infections typically cause abscesses (buildups of pus), ...

  4. Posttranscriptional modification of tRNA in thermophilic archaea (Archaebacteria).

    PubMed Central

    Edmonds, C G; Crain, P F; Gupta, R; Hashizume, T; Hocart, C H; Kowalak, J A; Pomerantz, S C; Stetter, K O; McCloskey, J A

    1991-01-01

    Nucleoside modification has been studied in unfractionated tRNA from 11 thermophilic archaea (archaebacteria), including phylogenetically diverse representatives of thermophilic methanogens and sulfur-metabolizing hyperthermophiles which grow optimally in the temperature range of 56 (Thermoplasma acidophilum) to 105 degrees C (Pyrodictium occultum), and for comparison from the most thermophilic bacterium (eubacterium) known, Thermotoga maritima (80 degrees C). Nine nucleosides are found to be unique to the archaea, six of which are structurally novel in being modified both in the base and by methylation in ribose and occur primarily in tRNA from the extreme thermophiles in the Crenarchaeota of the archaeal phylogenetic tree. 2-Thiothymine occurs in tRNA from Thermococcus sp., and constitutes the only known occurrence of the thymine moiety in archaeal RNA, in contrast to its near-ubiquitous presence in tRNA from bacteria and eukarya. A total of 33 modified nucleosides are rigorously characterized in archaeal tRNA in the present study, demonstrating that the structural range of posttranscriptional modifications in archaeal tRNA is more extensive than previously known. From a phylogenetic standpoint, certain tRNA modifications occur in the archaea which are otherwise unique to either the bacterial or eukaryal domain, although the overall patterns of modification are more typical of eukaryotes than bacteria. PMID:1708763

  5. Characterization of the arabinoxylan-degrading machinery of the thermophilic bacterium Herbinix hemicellulosilytica-Six new xylanases, three arabinofuranosidases and one xylosidase.

    PubMed

    Mechelke, M; Koeck, D E; Broeker, J; Roessler, B; Krabichler, F; Schwarz, W H; Zverlov, V V; Liebl, W

    2017-09-10

    Herbinix hemicellulosilytica is a newly isolated, gram-positive, anaerobic bacterium with extensive hemicellulose-degrading capabilities obtained from a thermophilic biogas reactor. In order to exploit its potential as a source for new industrial arabinoxylan-degrading enzymes, six new thermophilic xylanases, four from glycoside hydrolase family 10 (GH10) and two from GH11, three arabinofuranosidases (1x GH43, 2x GH51) and one β-xylosidase (GH43) were selected. The recombinantly produced enzymes were purified and characterized. All enzymes were active on different xylan-based polysaccharides and most of them showed temperature-vs-activity profiles with maxima around 55-65°C. HPAEC-PAD analysis of the hydrolysates of wheat arabinoxylan and of various purified xylooligosaccharides (XOS) and arabinoxylooligosaccharides (AXOS) was used to investigate their substrate and product specificities: among the GH10 xylanases, XynB showed a different product pattern when hydrolysing AXOS compared to XynA, XynC, and XynD. None of the GH11 xylanases was able to degrade any of the tested AXOS. All three arabinofuranosidases, ArfA, ArfB and ArfC, were classified as type AXH-m,d enzymes. None of the arabinofuranosidases was able to degrade the double-arabinosylated xylooligosaccharides XA 2+3 XX. β-Xylosidase XylA (GH43) was able to degrade unsubstituted XOS, but showed limited activity to degrade AXOS. Copyright © 2017. Published by Elsevier B.V.

  6. Thermosyntropha lipolytica gen. nov., sp. nov., a lipolytic, anaerobic, alkalitolerant, thermophilic bacterium utilizing short- and long-chain fatty acids in syntrophic coculture with a methanogenic archaeum.

    PubMed

    Svetlitshnyi, V; Rainey, F; Wiegel, J

    1996-10-01

    Three strains of an anaerobic thermophilic organoheterotrophic lipolytic alkalitolerant bacterium, Thermosyntropha lipolytica gen. nov., sp. nov. (type strain JW/VS-265T; DSM 11003), were isolated from alkaline hot springs of Lake Bogoria (Kenya). The cells were nonmotile, non-spore forming, straight or slightly curved rods. At 60 degrees C the pH range for growth determined at 25 degrees C [pH25 degrees C] was 7.15 to 9.5, with an optimum between 8.1 and 8.9 (pH60 degrees C of 7.6 and 8.1). At a pH25 degrees C of 8.5 the temperature range for growth was from 52 to 70 degrees C, with an optimum between 60 and 66 degrees C. The shortest doubling time was around 1 h. In pure culture the bacterium grew in a mineral base medium supplemented with yeast extract, tryptone, Casamino Acids, betaine, and crotonate as carbon sources, producing acetate as a major product and constitutively a lipase. During growth in the presence of olive oil, free long-chain fatty acids were accumulated in the medium but the pure culture could not utilize olive oil, triacylglycerols, short- and long-chain fatty acids, and glycerol for growth. In syntrophic coculture (Methanobacterium strain JW/VS-M29) the lipolytic bacteria grew on triacylglycerols and linear saturated and unsaturated fatty acids with 4 to 18 carbon atoms, but glycerol was not utilized. Fatty acids with even numbers of carbon atoms were degraded to acetate and methane, while from odd-numbered fatty acids 1 mol of propionate per mol of fatty acid was additionally formed. 16S rDNA sequence analysis identified Syntrophospora and Syntrophomonas spp. as closest phylogenetic neighbors.

  7. A family 5 β-mannanase from the thermophilic fungus Thielavia arenaria XZ7 with typical thermophilic enzyme features.

    PubMed

    Lu, Haiqiang; Zhang, Huitu; Shi, Pengjun; Luo, Huiying; Wang, Yaru; Yang, Peilong; Yao, Bin

    2013-09-01

    A novel β-mannanase gene, man5XZ7, was cloned from thermophilic fungus Thielavia arenaria XZ7, and successfully expressed in Pichia pastoris. The gene (1,110 bp) encodes a 369-amino acid polypeptide with a molecular mass of approximately 40.8 kDa. The deduced sequence of Man5XZ7 consists of a putative 17-residue signal peptide and a catalytic module belonging to glycoside hydrolase (GH) family 5, and displays 76 % identity with the experimentally verified GH 5 endo-β-1,4-mannanase from Podospora anserina. Recombinant Man5XZ7 was optimally active at 75 °C and pH 5.0 and exhibited high activity at a wide temperature range (>50.0 % activity at 50-85 °C). Moreover, it had good adaptability to acidic to basic pH (>74.1 % activity at pH 4.0-7.0 and 25.6 % even at pH 9.0) and good stability from pH 3.0 to 10.0. These enzymatic properties showed that Man5XZ7 was a new thermophilic and alkali-tolerant β-mannanase. Further amino acid composition analysis indicated that Man5XZ7 has several characteristic features of thermophilic enzymes.

  8. Fervidicola ferrireducens gen. nov., sp. nov., a thermophilic anaerobic bacterium from geothermal waters of the Great Artesian Basin, Australia.

    PubMed

    Ogg, Christopher D; Patel, Bharat K C

    2009-05-01

    A strictly anaerobic, thermophilic bacterium, designated strain Y170(T), was isolated from a microbial mat colonizing thermal waters of a run-off channel created by the free-flowing waters of a Great Artesian Basin (GAB) bore well (New Lorne bore; registered number 17263). Cells of strain Y170(T) were slightly curved rods (1.2-12x0.8-1.1 mum) and stained Gram-negative. The strain grew optimally in tryptone-yeast extract-glucose medium at 70 degrees C (temperature range for growth was 55-80 degrees C) and pH 7 (pH range for growth was 5-9). Strain Y170(T) grew poorly on yeast extract as a sole carbon source, but not on tryptone (0.2 %). Yeast extract could not be replaced by tryptone and was obligately required for growth on tryptone, peptone, glucose, fructose, galactose, cellobiose, mannose, sucrose, xylose, mannitol, formate, pyruvate, Casamino acids and threonine. No growth was observed on arabinose, lactose, maltose, raffinose, chitin, xylan, pectin, starch, acetate, benzoate, lactate, propionate, succinate, myo-inositol, ethanol, glycerol, amyl media, aspartate, leucine, glutamate, alanine, arginine, serine and glycine. End products detected from glucose fermentation were acetate, ethanol and presumably CO(2) and H(2). Iron(III), manganese(IV), thiosulfate and elemental sulfur, but not sulfate, sulfite, nitrate or nitrite, were used as electron acceptors in the presence of 0.2 % yeast extract. Iron(III) in the form of amorphous Fe(III) oxhydroxide and Fe(III) citrate was also reduced in the presence of tryptone, peptone and Casamino acids, but not with chitin, xylan, pectin, formate, starch, pyruvate, acetate, benzoate, threonine, lactate, propionate, succinate, inositol, ethanol, glycerol, mannitol, aspartate, leucine, glutamate, alanine, arginine, serine or glycine. Strain Y170(T) was not able to utilize molecular hydrogen and/or carbon dioxide in the presence or absence of iron(III). Chloramphenicol, streptomycin, tetracycline, penicillin and ampicillin and

  9. Effect of Recycle Sludge on Anaerobic Digestion of Palm Oil Mill Effluent in A Thermophilic Continuous Digester

    NASA Astrophysics Data System (ADS)

    Irvan; Trisakti, B.; Tomiuchi, Y.; Harahap, U.; Daimon, H.

    2017-06-01

    The objective of this research is to maintain short retention time and high degradation of palm oil mill effluent (POME) to biogas by applying recycle sludge. Fresh POME from Rambutan Mill without further treatment was used as feed. Two lab-scale digesters supported from Metawater Co. Ltd. have been applied to treat POME at thermophilic (55°C) condition. Both digesters were operated under intermittent operation mode. Experiments were performed in two methods: with and without recycle sludge. Hydraulic retention time (HRT) of both methods was maintained at 6 days, while sludge retention time (SRT) was maintained at various days. The result showed that by extending SRT in return sludge process where 25% of digested slurry recycled to the digester, improvement of volatile solid (VS) decomposition was obtained around 84% at HRT of 6 days and SRT of 21 days. Then, chemical oxygen demand (COD) removal efficiency could be reached until 85% by using recycle sludge.

  10. Characterization of wheat straw-degrading anaerobic alkali-tolerant mixed cultures from soda lake sediments by molecular and cultivation techniques

    PubMed Central

    Porsch, Katharina; Wirth, Balázs; Tóth, Erika M; Schattenberg, Florian; Nikolausz, Marcell

    2015-01-01

    Alkaline pretreatment has the potential to enhance the anaerobic digestion of lignocellulosic biomass to biogas. However, the elevated pH of the substrate may require alkalitolerant microbial communities for an effective digestion. Three mixed anaerobic lignocellulolytic cultures were enriched from sediments from two soda lakes with wheat straw as substrate under alkaline (pH 9) mesophilic (37°C) and thermophilic (55°C) conditions. The gas production of the three cultures ceased after 4 to 5 weeks, and the produced gas was composed of carbon dioxide and methane. The main liquid intermediates were acetate and propionate. The physiological behavior of the cultures was stable even after several transfers. The enrichment process was also followed by molecular fingerprinting (terminal restriction fragment length polymorphism) of the bacterial 16S rRNA gene and of the mcrA/mrtA functional gene for methanogens. The main shift in the microbial community composition occurred between the sediment samples and the first enrichment, whereas the structure was stable in the following transfers. The bacterial communities mainly consisted of Sphingobacteriales, Clostridiales and Spirochaeta, but differed at genus level. Methanothermobacter and Methanosarcina genera and the order Methanomicrobiales were predominant methanogenes in the obtained cultures. Additionally, single cellulolytic microorganisms were isolated from enrichment cultures and identified as members of the alkaliphilic or alkalitolerant genera. The results show that anaerobic alkaline habitats harbor diverse microbial communities, which can degrade lignocellulose effectively and are therefore a potential resource for improving anaerobic digestion. PMID:25737100

  11. ENVIRONMENTAL REGULATIONS AND TECHNOLOGY - AUTOTHERMAL THERMOPHILIC AEROBIC DIGESTION OF MUNICIPAL WASTEWATER SLUDGE

    EPA Science Inventory

    This document describes a promising technology — autothermal thermophilic aerobic digestion — for meeting the current and proposed U.S. federal requirements for pathogen controJ and land application of municipal wastewater sludge. Autothermal thermophilic aerobic digestion, or AT...

  12. Overcoming sodium toxicity by utilizing grass leaves as co-substrate during the start-up of batch thermophilic anaerobic digestion.

    PubMed

    Suwannoppadol, Suwat; Ho, Goen; Cord-Ruwisch, Ralf

    2012-12-01

    Sodium toxicity is a common problem causing inhibition of anaerobic digestion, and digesters treating highly concentrated wastes, such as food and municipal solid waste, and concentrated animal manure, are likely to suffer from partial or complete inhibition of methane-producing consortia, including methanogens. When grass clippings were added at the onset of anaerobic digestion of acetate containing a sodium concentration of 7.8 gNa(+)/L, a total methane production about 8L/L was obtained, whereas no methane was produced in the absence of grass leaves. In an attempt to narrow down which components of grass leaves caused decrease of sodium toxicity, different hypotheses were tested. Results revealed that betaine could be a significant compound in grass leaves causing reduction to sodium inhibition. Copyright © 2012 Elsevier Ltd. All rights reserved.

  13. Continuous thermal hydrolysis and energy integration in sludge anaerobic digestion plants.

    PubMed

    Fdz-Polanco, F; Velazquez, R; Perez-Elvira, S I; Casas, C; del Barrio, D; Cantero, F J; Fdz-Polanco, M; Rodriguez, P; Panizo, L; Serrat, J; Rouge, P

    2008-01-01

    A thermal hydrolysis pilot plant with direct steam injection heating was designed and constructed. In a first period the equipment was operated in batch to verify the effect of sludge type, pressure and temperature, residence time and solids concentration. Optimal operation conditions were reached for secondary sludge at 170 degrees C, 7 bar and 30 minutes residence time, obtaining a disintegration factor higher than 10, methane production increase by 50% and easy centrifugation In a second period the pilot plant was operated working with continuous feed, testing the efficiency by using two continuous anaerobic digester operating in the mesophilic and thermophilic range. Working at 12 days residence time, biogas production increases by 40-50%. Integrating the energy transfer it is possible to design a self-sufficient system that takes advantage of this methane increase to produce 40% more electric energy. (c) IWA Publishing 2008.

  14. Finding extraterrestrial sites for thermophiles.

    PubMed

    Naylor, T

    2004-04-01

    Virtually our entire knowledge of the universe comes from two sorts of measurement of the electromagnetic radiation from the stars and galaxies within it; either their flux through relatively wide bandpasses (photometry), or measurements of the shape and wavelength of relatively narrow lines via spectroscopy. These techniques are now being used to discover planets outside our solar system, and perhaps in the next 10 years will begin to characterize them. If a serious search is to be made for extraterrestrial thermophiles, we need predictions for the effects of thermophiles on their host planets that are observable with these techniques. In this paper I shall outline what sorts of observation are likely to be used in the next 15 years for extra-solar planet work. All of the journal articles quoted here can be found through http://adsabs.harvard.edu/abstract_service.html, and often also accessed as preprints at http://uk.arxiv.org/form/astro%20ph?MULTI=form%20+/-%20interface.

  15. Ethanol production by engineered thermophiles.

    PubMed

    Olson, Daniel G; Sparling, Richard; Lynd, Lee R

    2015-06-01

    We compare a number of different strategies that have been pursued to engineer thermophilic microorganisms for increased ethanol production. Ethanol production from pyruvate can proceed via one of four pathways, which are named by the key pyruvate dissimilating enzyme: pyruvate decarboxylase (PDC), pyruvate dehydrogenase (PDH), pyruvate formate lyase (PFL), and pyruvate ferredoxin oxidoreductase (PFOR). For each of these pathways except PFL, we see examples where ethanol production has been engineered with a yield of >90% of the theoretical maximum. In each of these cases, this engineering was achieved mainly by modulating expression of native genes. We have not found an example where a thermophilic ethanol production pathway has been transferred to a non-ethanol-producing organism to produce ethanol at high yield. A key reason for the lack of transferability of ethanol production pathways is the current lack of understanding of the enzymes involved. Copyright © 2015 Elsevier Ltd. All rights reserved.

  16. Gas analysis reveals novel aerobic deammonification in thermophilic aerobic digestion.

    PubMed

    Yi, Y S; Kim, S; An, S; Choi, S I; Choi, E; Yun, Z

    2003-01-01

    A laboratory-scale thermophilic aerobic digester was operated with piggery wastewater. The operating temperature varied from 50-70 degrees C. It has been found that excessive nitrogen removal occurred in the laboratory-scale thermophilic system at various HRTs. Nitrite and nitrate were not observed in the effluent. Gas measurement reveals the presence of significant amount of N2O along with NH3 gas. The rational production of N2O gas in accordance with temperature and HRT suggests that biologically mediated deammonification processes significantly contribute to the N removal. Although further microbiological investigation is required to clarify the exact nitrogen removal mechanism, the large production of N2O gas seems to be a result of the existence of a rapid growing heterotrophic deammonification process in the thermophilic system.

  17. Biohydrogen Production by the Thermophilic Bacterium Caldicellulosiruptor saccharolyticus: Current Status and Perspectives

    PubMed Central

    Bielen, Abraham A. M.; Verhaart, Marcel R. A.; van der Oost, John; Kengen, Servé W. M.

    2013-01-01

    Caldicellulosiruptor saccharolyticus is one of the most thermophilic cellulolytic organisms known to date. This Gram-positive anaerobic bacterium ferments a broad spectrum of mono-, di- and polysaccharides to mainly acetate, CO2 and hydrogen. With hydrogen yields approaching the theoretical limit for dark fermentation of 4 mol hydrogen per mol hexose, this organism has proven itself to be an excellent candidate for biological hydrogen production. This review provides an overview of the research on C. saccharolyticus with respect to the hydrolytic capability, sugar metabolism, hydrogen formation, mechanisms involved in hydrogen inhibition, and the regulation of the redox and carbon metabolism. Analysis of currently available fermentation data reveal decreased hydrogen yields under non-ideal cultivation conditions, which are mainly associated with the accumulation of hydrogen in the liquid phase. Thermodynamic considerations concerning the reactions involved in hydrogen formation are discussed with respect to the dissolved hydrogen concentration. Novel cultivation data demonstrate the sensitivity of C. saccharolyticus to increased hydrogen levels regarding substrate load and nitrogen limitation. In addition, special attention is given to the rhamnose metabolism, which represents an unusual type of redox balancing. Finally, several approaches are suggested to improve biohydrogen production by C. saccharolyticus. PMID:25371332

  18. Removal of slowly biodegradable COD in combined thermophilic UASB and MBBR systems.

    PubMed

    Ji, M; Yu, J; Chen, H; Yue, P L

    2001-09-01

    Starch, cellulose and polyvinyl alcohol (PVA) are common substrates of the slowly biodegradable COD (SBCOD) in industrial wastewaters. Removal of the individual and mixed SbCOD substrates was investigated in a combined system of thermophilic upflow anaerobic sludge blanket (TUASB) reactor (55 degrees C) and aerobic moving bed biofilm reactor (MBBR). The removal mechanisms of the three SBCOD substrates were quite different. Starch-COD was almost equally utilized and removed in the two reactors. Cellulose-COD was completely (97-98%) removed from water in the TUASB reactor by microbial entrapment and sedimentation of the cellulose fibers. PVA alone was hardly biodegraded and removed by the combined reactors. However, PVA-COD could be removed to some extent in a binary solution of starch (77%) plus PVA (23%). The PVA macromolecules in the binary solution actually affected the microbial activity in the TUASB reactor resulting accumulation of volatile fatty acids, which shifted the overall COD removal from the TUASB to the MBBR reactor where SBCOD including PVA-COD was removed. Since the three SBCOD substrates were removed by different mechanisms, the combined reactors showed a better and more stable performance than individual reactors.

  19. Carboxydothermus islandicus sp. nov., a thermophilic, hydrogenogenic, carboxydotrophic bacterium isolated from a hot spring.

    PubMed

    Novikov, Andrey A; Sokolova, Tatyana G; Lebedinsky, Alexander V; Kolganova, Tatyana V; Bonch-Osmolovskaya, Elizaveta A

    2011-10-01

    An anaerobic, thermophilic bacterium, strain SET IS-9(T), was isolated from an Icelandic hot spring. Cells of strain SET IS-9(T) are short, slightly curved, motile rods. The strain grows chemolithotrophically on CO, producing equimolar quantities of H(2) and CO(2). It also grows fermentatively on lactate or pyruvate in the presence of yeast extract (0.2 g l(-1)). Products of pyruvate fermentation are acetate, CO(2) and H(2). Growth occurs at 50-70 °C, with an optimum at 65 °C, and at pH 5.0-8.0, with an optimum at pH 5.5-6.0. The generation time during chemolithotrophic growth on CO under optimal conditions is 2.0 h. 16S rRNA gene sequence analysis suggested that the organism belongs to the genus Carboxydothermus. On the basis of phenotypic features and phylogenetic analysis, Carboxydothermus islandicus sp. nov. is proposed, with the type strain SET IS-9(T) ( = DSM 21830(T)  = VKM B-2561(T)). An emended description of the genus Carboxydothermus is also given.

  20. Cloning, Expression and Characterization of a Novel Thermophilic Polygalacturonase from Caldicellulosiruptor bescii DSM 6725

    PubMed Central

    Chen, Yanyan; Sun, Dejun; Zhou, Yulai; Liu, Liping; Han, Weiwei; Zheng, Baisong; Wang, Zhi; Zhang, Zuoming

    2014-01-01

    We cloned the gene ACM61449 from anaerobic, thermophilic Caldicellulosiruptor bescii, and expressed it in Escherichia coli origami (DE3). After purification through thermal treatment and Ni-NTA agarose column extraction, we characterized the properties of the recombinant protein (CbPelA). The optimal temperature and pH of the protein were 72 °C and 5.2, respectively. CbPelA demonstrated high thermal-stability, with a half-life of 14 h at 70 °C. CbPelA also showed very high activity for polygalacturonic acid (PGA), and released monogalacturonic acid as its sole product. The Vmax and Km of CbPelA were 384.6 U·mg−1 and 0.31 mg·mL−1, respectively. CbPelA was also able to hydrolyze methylated pectin (48% and 10% relative activity on 20%–34% and 85% methylated pectin, respectively). The high thermo-activity and methylated pectin hydrolization activity of CbPelA suggest that it has potential applications in the food and textile industry. PMID:24705464

  1. Comparative genomic analysis of the thermophilic biomass-degrading fungi Myceliophthora thermophila and Thielavia terrestris

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Berka, Randy M.; Grigoriev, Igor V.; Otillar, Robert

    2011-10-02

    Thermostable enzymes and thermophilic cell factories may afford economic advantages in the production of many chemicals and biomass-based fuels. Here we describe and compare the genomes of two thermophilic fungi, Myceliophthora thermophila and Thielavia terrestris. To our knowledge, these genomes are the first described for thermophilic eukaryotes and the first complete telomere-to-telomere genomes for filamentous fungi. Genome analyses and experimental data suggest that both thermophiles are capable of hydrolyzing all major polysaccharides found in biomass. Examination of transcriptome data and secreted proteins suggests that the two fungi use shared approaches in the hydrolysis of cellulose and xylan but distinct mechanismsmore » in pectin degradation. Characterization of the biomass-hydrolyzing activity of recombinant enzymes suggests that these organisms are highly efficient in biomass decomposition at both moderate and high temperatures. Furthermore, we present evidence suggesting that aside from representing a potential reservoir of thermostable enzymes, thermophilic fungi are amenable to manipulation using classical and molecular genetics.« less

  2. Comparative genomic analysis of the thermophilic biomass-degrading fungi Myceliophthora thermophila and Thielavia terrestris

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Berka, Randy M.; Grigoriev, Igor V.; Otillar, Robert

    2011-05-16

    Thermostable enzymes and thermophilic cell factories may afford economic advantages in the production of many chemicals and biomass-based fuels. Here we describe and compare the genomes of two thermophilic fungi, Myceliophthora thermophila and Thielavia terrestris. To our knowledge, these genomes are the first described for thermophilic eukaryotes and the first complete telomere-to-telomere genomes for filamentous fungi. Genome analyses and experimental data suggest that both thermophiles are capable of hydrolyzing all major polysaccharides found in biomass. Examination of transcriptome data and secreted proteins suggests that the two fungi use shared approaches in the hydrolysis of cellulose and xylan but distinct mechanismsmore » in pectin degradation. Characterization of the biomass-hydrolyzing activity of recombinant enzymes suggests that these organisms are highly efficient in biomass decomposition at both moderate and high temperatures. Furthermore, we present evidence suggesting that aside from representing a potential reservoir of thermostable enzymes, thermophilic fungi are amenable to manipulation using classical and molecular genetics.« less

  3. Utilization of solid and liquid waste generated during ethanol fermentation process for production of gaseous fuel through anaerobic digestion--a zero waste approach.

    PubMed

    Narra, Madhuri; Balasubramanian, Velmurugan

    2015-03-01

    Preliminary investigations were performed in the laboratory using batch reactors at 10% solid concentration for the assessment of the biogas production at thermophilic and mesophilic temperatures using solid residues generated during ethanol fermentation process. One kg of solid residues (left after enzyme extraction and enzymatic hydrolysis) from thermophilic reactors (TR1 and TR2) produced around 131 and 84L of biogas, respectively, whereas biogas production from mesophilic reactors (MR1 and MR2) was 86 and 62L, respectively. After 20 and 35days of retention time, the TS and VS reductions from TR1, TR2 and MR1, MR2 were found to be 39.2% and 35.0%, 67.3% and 61.0%, 21.0% and 18.0%, 34.7% and 27.8%, respectively. Whereas the liquid waste was treated using four laboratory anaerobic hybrid reactors (AHRs) with two different natural and synthetic packing media at 15-3days HRTs. AHRs packed with natural media showed better COD removal efficiency and methane yield. Copyright © 2015 Elsevier Ltd. All rights reserved.

  4. Comparison of methods for isolation and enumeration of thermophilic actinomycetes from dust.

    PubMed Central

    Treuhaft, M W; Arden Jones, M P

    1982-01-01

    Thermophilic actinomycetes are the primary sensitizing agents in farmer's lung disease. We compared dilution pour-plate and spread-plate methods for their usefulness in enumerating thermophilic actinomycetes in moldy silage dust and evaluated the ability of a nonquantitative gravity settling technique to recover thermophilic actinomycetes from moldy silage. Spread plates and pour plates yielded similar estimates of total thermophiles. Higher counts were observed on spread plates (P less than 0.05) for Thermoactinomyces candidus, Micropolyspora faeni, and Saccharomonospora viridis. M. faeni and S. viridis were less efficient than T. candidus in breaking through the agar of pour plates to form colonies which could be identified. Coefficients of variability were less than 10% for the two methods. The relative proportion of organisms recovered by the settling method correlated well with that recovered on spread plates for M. faeni (r = 0.79), S. viridis (r = 0.88), and Thermomonospora spp. (r = 0.79), but not well for T. candidus (r = 0.28). When sophisticated air-sampling equipment is not available, dilution spread plates of dust washings provide a reproducible method for enumerating a broad range of thermophilic actinomycetes of interest. The gravity settling method is a simple, rapid alternative when isolation is all that is required. PMID:6761363

  5. Conventional and thermophilic aerobic treatability of high strength oily pet food wastewater using membrane-coupled bioreactors.

    PubMed

    Kurian, R; Acharya, C; Nakhla, G; Bassi, A

    2005-11-01

    Although thermophilic treatment systems have recently gained considerable interest, limited information exists on the comparative performances of membrane-coupled bioreactors (MBR) at thermophilic and conventional conditions. In this study aerobic MBRs operating at room temperature (20 degrees C) and at lower thermophilic range (45 degrees C) were investigated for the treatment of dissolved air flotation (DAF) pretreated pet food wastewater. The particular wastewater is characterized by oil and grease (O & G) concentrations as high as 6 g/L, COD of 51 g/L, BOD of 16 g/L and volatile fatty acid (VFA) of 8.3 g/L. The performances of the two systems in terms of COD, BOD and O & G removal at varying hydraulic retention time (HRT) are compared. COD removal efficiencies in the thermophilic MBR varied from 75% to 98% and remained constant at 94% in the conventional MBR. The O & G removal efficiencies were 66-86% and 98% in the thermophilic and conventional MBR, respectively. Interestingly, high concentrations of VFA were recorded, equivalent to 50-73% of total COD, in the thermophilic MBR effluent. The observed yield in the thermophilic MBR was 40% of that observed in the conventional MBR.

  6. Genome sequence of the moderately thermophilic sulfur-reducing bacterium Thermanaerovibrio velox type strain (Z-9701T) and emended description of the genus Thermanaerovibrio

    PubMed Central

    Palaniappan, Krishna; Meier-Kolthoff, Jan P.; Teshima, Hazuki; Nolan, Matt; Lapidus, Alla; Tice, Hope; Del Rio, Tijana Glavina; Cheng, Jan-Fang; Han, Cliff; Tapia, Roxanne; Goodwin, Lynne A.; Pitluck, Sam; Liolios, Konstantinos; Mavromatis, Konstantinos; Pagani, Ioanna; Ivanova, Natalia; Mikhailova, Natalia; Pati, Amrita; Chen, Amy; Rohde, Manfred; Mayilraj, Shanmugam; Spring, Stefan; Detter, John C.; Göker, Markus; Bristow, James; Eisen, Jonathan A.; Markowitz, Victor; Hugenholtz, Philip; Kyrpides, Nikos C.; Klenk, Hans-Peter; Woyke, Tanja

    2013-01-01

    Thermanaerovibrio velox Zavarzina et al. 2000 is a member of the Synergistaceae, a family in the phylum Synergistetes that is already well-characterized at the genome level. Members of this phylum were described as Gram-negative staining anaerobic bacteria with a rod/vibrioid cell shape and possessing an atypical outer cell envelope. They inhabit a large variety of anaerobic environments including soil, oil wells, wastewater treatment plants and animal gastrointestinal tracts. They are also found to be linked to sites of human diseases such as cysts, abscesses, and areas of periodontal disease. The moderately thermophilic and organotrophic T. velox shares most of its morphologic and physiologic features with the closely related species, T. acidaminovorans. In addition to Su883T, the type strain of T. acidaminovorans, stain Z-9701T is the second type strain in the genus Thermanaerovibrio to have its genome sequence published. Here we describe the features of this organism, together with the non-contiguous genome sequence and annotation. The 1,880,838 bp long chromosome (non-contiguous finished sequence) with its 1,751 protein-coding and 59 RNA genes is a part of the Genomic Encyclopedia of Bacteria and Archaea project. PMID:24501645

  7. Application of high rate, high temperature anaerobic digestion to fungal thermozyme hydrolysates from carbohydrate wastes.

    PubMed

    Forbes, C; O'Reilly, C; McLaughlin, L; Gilleran, G; Tuohy, M; Colleran, E

    2009-05-01

    The objective of this study was to examine the feasibility of using a two-step, fully biological and sustainable strategy for the treatment of carbohydrate rich wastes. The primary step in this strategy involves the application of thermostable enzymes produced by the thermophilic, aerobic fungus, Talaromyces emersonii, to carbohydrate wastes producing a liquid hydrolysate discharged at elevated temperatures. To assess the potential of thermophilic treatment of this hydrolysate, a comparative study of thermophilic and mesophilic digestion of four sugar rich thermozyme hydrolysate waste streams was conducted by operating two high rate upflow anaerobic hybrid reactors (UAHR) at 37 degrees C (R1) and 55 degrees C (R2). The operational performance of both reactors was monitored from start-up by assessing COD removal efficiencies, volatile fatty acid (VFA) discharge and % methane of the biogas produced. Rapid start-up of both R1 and R2 was achieved on an influent composed of the typical sugar components of the organic fraction of municipal solid waste (OFMSW). Both reactors were subsequently challenged in terms of volumetric loading rate (VLR) and it was found that a VLR of 9 gCOD l(-1)d(-1) at a hydraulic retention time (HRT) of 1 day severely affected the thermophilic reactor with instability characterised by a build up of volatile fatty acid (VFA) intermediates in the effluent. The influent to both reactors was changed to a simple glucose and sucrose-based influent supplied at a VLR of 4.5 gCOD l(-1)d(-1) and HRT of 2 days prior to the introduction of thermozyme hydrolysates. Four unique thermozyme hydrolysates were subsequently supplied to the reactors, each for a period of 10 HRTs. The applied hydrolysates were derived from apple pulp, bread, carob powder and cardboard, all of which were successfully and comparably converted by both reactors. The % total carbohydrate removal by both reactors was monitored during the application of the sugar rich thermozyme

  8. Optimal subset selection of primary sequence features using the genetic algorithm for thermophilic proteins identification.

    PubMed

    Wang, LiQiang; Li, CuiFeng

    2014-10-01

    A genetic algorithm (GA) coupled with multiple linear regression (MLR) was used to extract useful features from amino acids and g-gap dipeptides for distinguishing between thermophilic and non-thermophilic proteins. The method was trained by a benchmark dataset of 915 thermophilic and 793 non-thermophilic proteins. The method reached an overall accuracy of 95.4 % in a Jackknife test using nine amino acids, 38 0-gap dipeptides and 29 1-gap dipeptides. The accuracy as a function of protein size ranged between 85.8 and 96.9 %. The overall accuracies of three independent tests were 93, 93.4 and 91.8 %. The observed results of detecting thermophilic proteins suggest that the GA-MLR approach described herein should be a powerful method for selecting features that describe thermostabile machines and be an aid in the design of more stable proteins.

  9. Enzyme activity screening of thermophilic bacteria isolated from Dusun Tua Hot Spring, Malaysia

    NASA Astrophysics Data System (ADS)

    Msarah, Marwan; Ibrahim, Izyanti; Aqma, Wan Syaidatul

    2018-04-01

    Thermophilic bacteria have biotechnological importance due to the availability of unique enzymes which are stable in extreme circumstances. The aim of this study includes to isolate thermophilic bacteria from hot spring and screen for important enzyme activities. Water samples from the Dusun Tua Hot Spring were collected and the physiochemical characterisation of water was measured. Eight thermophilic bacteria were isolated and determined to have at least three strong enzyme activity including protease, lipase, amylase, cellulase, pectinase and xylanase. The results showed that HuluC2 displayed all the enzyme activities and can be further studied.

  10. Fast production of methane by anaerobic digestion. Annual progress report, May 24, 1976--May 23, 1977

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Finney, C.D.; Evans II, R.S.; Finney, K.A.

    1977-06-01

    Since the productional cost of methane generated by anaerobic digestion of cellulose is on the economic borderline and the cost could be reduced by increasing the rate of the digestion process, a research program was undertaken to delineate the most promising areas of development. The concept that the step involving transfer of products from solution is rate-limiting and inhibiting in anaerobic digestion was supported by all evidence available. The most significant design implication of this concept is that faster gas production can be achieved in a two-stage digestion system in which unreactive solids are eliminated after the hydrolysis step somore » that the effluent to the gas-producing stage possesses a low viscosity. The advantages and disadvantages of three hydrolysis methods (enzymatic, anaerobic, and acid) are reviewed.« less

  11. Anaerobic hydrogen production from unhydrolyzed mushroom farm waste by indigenous microbiota.

    PubMed

    Lin, Chiu-Yue; Lay, Chyi-How; Sung, I-Yuan; Sen, Biswarup; Chen, Chin-Chao

    2017-10-01

    The cultivation of mushrooms generates large amounts of waste polypropylene bags stuffed with wood flour and bacterial nutrients that makes the mushroom waste (MW) a potential feedstock for anaerobic bioH 2 fermentation. MW indigenous bacteria were enriched using thermophilic temperature (55°C) for use as the seed inoculum without any external seeding. The peak hydrogen production rate (6.84 mmol H 2 /L-d) was obtained with cultivation pH 8 and substrate concentration of 60 g MW/L in batch fermentation. Hydrogen production yield (HY) is pH and substrate concentration dependent with an HY decline occurring at pH and substrate concentration increasing from pH 8 to 10 and 60 to 80 g MW/L, respectively. The fermentation bioH 2 production from MW is in an acetate-type metabolic path. Copyright © 2017 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.

  12. Radioisotopic, Culture-Based, and Oligonucleotide Microchip Analyses of Thermophilic Microbial Communities in a Continental High-Temperature Petroleum Reservoir†

    PubMed Central

    Bonch-Osmolovskaya, Elizaveta A.; Miroshnichenko, Margarita L.; Lebedinsky, Alexander V.; Chernyh, Nikolai A.; Nazina, Tamara N.; Ivoilov, Valery S.; Belyaev, Sergey S.; Boulygina, Eugenia S.; Lysov, Yury P.; Perov, Alexander N.; Mirzabekov , Andrei D.; Hippe, Hans; Stackebrandt, Erko; L'Haridon, Stéphane; Jeanthon, Christian

    2003-01-01

    Activity measurements by radioisotopic methods and cultural and molecular approaches were used in parallel to investigate the microbial biodiversity and its physiological potential in formation waters of the Samotlor high-temperature oil reservoir (Western Siberia, Russia). Sulfate reduction with rates not exceeding 20 nmol of H2S liter−1 day−1 occurred at 60 and 80°C. In upper horizons (AB, A, and B), methanogenesis (lithotrophic and/or acetoclastic) was detected only in wells in which sulfate reduction did not occur. In some of the wells from deeper (J) horizons, high-temperature sulfate reduction and methanogenesis occurred simultaneously, the rate of lithotrophic methanogenesis exceeding 80 nmol of CH4 liter−1 day−1. Enrichment cultures indicated the presence of diverse physiological groups representing aerobic and anaerobic thermophiles and hyperthermophiles; fermentative organotrophs were predominant. Phylogenetic analyses of 15 isolates identified representatives of the genera Thermotoga, Thermoanaerobacter, Geobacillus, Petrotoga, Thermosipho, and Thermococcus, the latter four being represented by new species. Except for Thermosipho, the isolates were members of genera recovered earlier from similar habitats. DNA obtained from three samples was hybridized with a set of oligonucleotide probes targeting selected microbial groups encompassing key genera of thermophilic bacteria and archaea. Oligonucleotide microchip analyses confirmed the cultural data but also revealed the presence of several groups of microorganisms that escaped cultivation, among them representatives of the Aquificales/Desulfurobacterium-Thermovibrio cluster and of the genera Desulfurococcus and Thermus, up to now unknown in this habitat. The unexpected presence of these organisms suggests that their distribution may be much wider than suspected. PMID:14532074

  13. [Anaerobic bacteria isolated from patients with suspected anaerobic infections].

    PubMed

    Ercis, Serpil; Tunçkanat, Ferda; Hasçelik, Gülşen

    2005-10-01

    The study involved 394 clinical samples sent to the Clinical Microbiology Laboratory of Hacettepe University Adult Hospital between January 1997 and May 2004 for anaerobic cultivation. Since multiple cultures from the same clinical samples of the same patient were excluded, the study was carried on 367 samples. The anaerobic cultures were performed in anaerobic jar using AnaeroGen kits (Oxoid, Basingstoke, U.K.) or GENbox (bioMérieux, Lyon, France). The isolates were identified by both classical methods and "BBL Crystal System" (Becton Dickinson, U.S.A.). While no growth was detected in 120 (32.7%) of the clinical samples studied, in 144 samples (39.2%) only aerobes, in 28 (7.6%) only anaerobes and in 75 (20.5%) of the samples both aerobes and anaerobes were isolated. The number of the anaerobic isolates was 217 from 103 samples with anaerobic growth. Of these 103 samples 15 showed single bacterial growth whereas in 88 samples multiple bacterial isolates were detected. Anaerobic isolates consisted of 92 Gram negative bacilli (Bacteroides spp. 50, Prevotella spp. 14, Porphyromonas spp. 10, Fusobacterium spp. 7, Tisierella spp. 2, unidentified 9), 57 Gram positive bacilli (Clostridium spp.17, Propionibacterium spp. 16, Lactobacillus spp. 8, Actinomyces spp. 5, Eubacterium spp. 2, Bifidobacterium adolescentis 1, Mobiluncus mulieris 1, unidentified nonspore forming rods 7), 61 Gram positive cocci (anaerobic cocci 44, microaerophilic cocci 17), and 7 Gram negative cocci (Veillonella spp.). In conclusion, in the samples studied with prediagnosis of anaerobic infection, Bacteroides spp. (23%) were the most common bacteria followed by anaerobic Gram positive cocci (20.3%) and Clostridium spp (7.8%).

  14. Bacterial community structure transformed after thermophilically composting human waste in Haiti

    PubMed Central

    Kramer, Sasha; Roy, Monika; Reid, Francine C.; Dubinsky, Eric A.

    2017-01-01

    Recycling human waste for beneficial use has been practiced for millennia. Aerobic (thermophilic) composting of sewage sludge has been shown to reduce populations of opportunistically pathogenic bacteria and to inactivate both Ascaris eggs and culturable Escherichia coli in raw waste, but there is still a question about the fate of most fecal bacteria when raw material is composted directly. This study undertook a comprehensive microbial community analysis of composting material at various stages collected over 6 months at two composting facilities in Haiti. The fecal microbiota signal was monitored using a high-density DNA microarray (PhyloChip). Thermophilic composting altered the bacterial community structure of the starting material. Typical fecal bacteria classified in the following groups were present in at least half the starting material samples, yet were reduced below detection in finished compost: Prevotella and Erysipelotrichaceae (100% reduction of initial presence), Ruminococcaceae (98–99%), Lachnospiraceae (83–94%, primarily unclassified taxa remained), Escherichia and Shigella (100%). Opportunistic pathogens were reduced below the level of detection in the final product with the exception of Clostridium tetani, which could have survived in a spore state or been reintroduced late in the outdoor maturation process. Conversely, thermotolerant or thermophilic Actinomycetes and Firmicutes (e.g., Thermobifida, Bacillus, Geobacillus) typically found in compost increased substantially during the thermophilic stage. This community DNA-based assessment of the fate of human fecal microbiota during thermophilic composting will help optimize this process as a sanitation solution in areas where infrastructure and resources are limited. PMID:28570610

  15. Presence of Thermophilic Bacteria in Laundry and Domestic Hot-Water Heaters

    PubMed Central

    Brock, Thomas D.; Boylen, Kathryn L.

    1973-01-01

    Thermophilic bacteria resembling Thermus aquaticus were isolated from hot water taken from domestic and commercial hot-water tanks. Cold water from the same locations never yielded thermophilic bacteria, suggesting that the bacteria were growing in the tanks. In contrast to the T. aquaticus isolates from hot springs, the present isolates were rarely pigmented. In general, the hotter sources more frequently yielded bacteria. PMID:4568892

  16. Methane production by anaerobic digestion of Bermuda grass

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Klass, D.L.; Ghosh, S.

    1981-01-01

    Bermuda grass (Cynodon dactylon) is one of the high-yield warm-season grasses that has been suggested as a promising raw material for conversion to methane. Experimental work performed with laboratory digesters to study the anaerobic digestion of Coastal Bermuda grass harvested in Louisiana and having a C/N ratio of 24 is described. Methane yields of about 1.9 SCF/lb of volatile solids (VS) added were observed under conventional mesophilic high-rate conditions. When supplemental nitrogen additions were made, the methane yields increased. This observation along with the compositional data compiled on the grass used in this work indicated that the nitrogen content ofmore » the unsupplemented grass was insufficient to sustain high-rate digestion at the higher yield level. However, as the C/N ratio was reduced by addition of ammonium chloride, the methane yield continually increased up to 3.5 SCF/lb added at the lowest C/N ratio examined (6.3) even after relatively high concentrations of ammonium nitrogen were measured in the effluent. It appears that the added nutrient had a stimulatory effect on methane production above the point where nitrogen was not limiting. Thermophilic digestion with supplemental nitrogen additions afforded methane yields of about 2.7 SCF/lb VS added. Carbon and energy balances were calculated and the relative biodegradabilities of the organics were estimated. It was concluded from this work that Coastal Bermuda grass can be converted to high-methane gas under conventional anaerobic digestion conditions. The performance of the particular lot of grass studied was substantially improved by supplemental nitrogen additions. (Refs. 12).« less

  17. The performance and microbial diversity of temperature-phased hyperthermophilic and thermophilic anaerobic digestion system fed with organic waste.

    PubMed

    Lee, M Y; Cheon, J H; Hidaka, T; Tsuno, H

    2008-01-01

    The objective of this study was to evaluate the performances and microbial diversities for development of the effective hyperthermophilic digester system that consists of a hyperthermophilic reactor and hyperthermophilic or thermophilic reactor in series. Lab-scale reactors were operated continuously fed with artificial kitchen garbage. The effect of temperature on the acidification step was firstly investigated. Results indicated that 20.8% of COD solubilization was achieved at 70 degrees C, with 12.6% at 80 degrees C. The average protein solubilization reached 31% at 80 degrees C. Methane conversion efficiency following the acidification was around 85% on average at 55 degrees C, but decreased with increasing temperature and methane gas was not produced over 73 degrees C. As well, bacteria affiliated with the methanogens dominated the population below 65 degrees C, while those affiliated with acidogens were predominant over 73 degrees C. These results indicated that the hyperthermophilic process has considerable benefits to treat wastewater or waste containing high concentration of protein.

  18. Fixation of CO2 and CO on a diverse range of carbohydrates using anaerobic, non-photosynthetic mixotrophy.

    PubMed

    Maru, Biniam T; Munasinghe, Pradeep C; Gilary, Hadar; Jones, Shawn W; Tracy, Bryan P

    2018-04-01

    Biological CO2 fixation is an important technology that can assist in combating climate change. Here, we show an approach called anaerobic, non-photosynthetic mixotrophy can result in net CO2 fixation when using a reduced feedstock. This approach uses microbes called acetogens that are capable of concurrent utilization of both organic and inorganic substrates. In this study, we investigated the substrate utilization of 17 different acetogens, both mesophilic and thermophilic, on a variety of different carbohydrates and gases. Compared to most model acetogen strains, several non-model mesophilic strains displayed greater substrate flexibility, including the ability to utilize disaccharides, glycerol and an oligosaccharide, and growth rates. Three of these non-model strains (Blautia producta, Clostridium scatologenes and Thermoanaerobacter kivui) were chosen for further characterization, under a variety of conditions including H2- or syngas-fed sugar fermentations and a CO2-fed glycerol fermentation. In all cases, CO2 was fixed and carbon yields approached 100%. Finally, the model acetogen C. ljungdahlii was engineered to utilize glucose, a non-preferred sugar, while maintaining mixotrophic behavior. This work demonstrates the flexibility and robustness of anaerobic, non-photosynthetic mixotrophy as a technology to help reduce CO2 emissions.

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

    PubMed Central

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

    2014-01-01

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

  20. Anaerobic co-digestion of winery waste and waste activated sludge: assessment of process feasibility.

    PubMed

    Da Ros, C; Cavinato, C; Cecchi, F; Bolzonella, D

    2014-01-01

    In this study the anaerobic co-digestion of wine lees together with waste activated sludge in mesophilic and thermophilic conditions was tested at pilot scale. Three organic loading rates (OLRs 2.8, 3.3 and 4.5 kgCOD/m(3)d) and hydraulic retention times (HRTs 21, 19 and 16 days) were applied to the reactors, in order to evaluate the best operational conditions for the maximization of the biogas yields. The addition of lee to sludge determined a higher biogas production: the best yield obtained was 0.40 Nm(3)biogas/kgCODfed. Because of the high presence of soluble chemical oxygen demand (COD) and polyphenols in wine lees, the best results in terms of yields and process stability were obtained when applying the lowest of the three organic loading rates tested together with mesophilic conditions.

  1. Metagenomics of Thermophiles with a Focus on Discovery of Novel Thermozymes

    PubMed Central

    DeCastro, María-Eugenia; Rodríguez-Belmonte, Esther; González-Siso, María-Isabel

    2016-01-01

    Microbial populations living in environments with temperatures above 50°C (thermophiles) have been widely studied, increasing our knowledge in the composition and function of these ecological communities. Since these populations express a broad number of heat-resistant enzymes (thermozymes), they also represent an important source for novel biocatalysts that can be potentially used in industrial processes. The integrated study of the whole-community DNA from an environment, known as metagenomics, coupled with the development of next generation sequencing (NGS) technologies, has allowed the generation of large amounts of data from thermophiles. In this review, we summarize the main approaches commonly utilized for assessing the taxonomic and functional diversity of thermophiles through metagenomics, including several bioinformatics tools and some metagenome-derived methods to isolate their thermozymes. PMID:27729905

  2. Changes in the antibiotic susceptibility of anaerobic bacteria from 2007-2009 to 2010-2012 based on the CLSI methodology.

    PubMed

    Hastey, Christine J; Boyd, Halsey; Schuetz, Audrey N; Anderson, Karen; Citron, Diane M; Dzink-Fox, Jody; Hackel, Meredith; Hecht, David W; Jacobus, Nilda V; Jenkins, Stephen G; Karlsson, Maria; Knapp, Cynthia C; Koeth, Laura M; Wexler, Hannah; Roe-Carpenter, Darcie E

    2016-12-01

    Antimicrobial susceptibility testing of anaerobic isolates was conducted at four independent sites from 2010 to 2012 and compared to results from three sites during the period of 2007-2009. This data comparison shows significant changes in antimicrobial resistance in some anaerobic groups. Therefore, we continue to recommend institutions regularly perform susceptibility testing when anaerobes are cultured from pertinent sites. Annual generation of an institutional-specific antibiogram is recommended for tracking of resistance trends over time. Copyright © 2016 Elsevier Ltd. All rights reserved.

  3. Potential and utilization of thermophiles and thermostable enzymes in biorefining

    PubMed Central

    Turner, Pernilla; Mamo, Gashaw; Karlsson, Eva Nordberg

    2007-01-01

    In today's world, there is an increasing trend towards the use of renewable, cheap and readily available biomass in the production of a wide variety of fine and bulk chemicals in different biorefineries. Biorefineries utilize the activities of microbial cells and their enzymes to convert biomass into target products. Many of these processes require enzymes which are operationally stable at high temperature thus allowing e.g. easy mixing, better substrate solubility, high mass transfer rate, and lowered risk of contamination. Thermophiles have often been proposed as sources of industrially relevant thermostable enzymes. Here we discuss existing and potential applications of thermophiles and thermostable enzymes with focus on conversion of carbohydrate containing raw materials. Their importance in biorefineries is explained using examples of lignocellulose and starch conversions to desired products. Strategies that enhance thermostablity of enzymes both in vivo and in vitro are also assessed. Moreover, this review deals with efforts made on developing vectors for expressing recombinant enzymes in thermophilic hosts. PMID:17359551

  4. Combining high-rate aerobic wastewater treatment with anaerobic digestion of waste activated sludge at a pulp and paper mill.

    PubMed

    Magnusson, Björn; Ekstrand, Eva-Maria; Karlsson, Anna; Ejlertsson, Jörgen

    2018-05-01

    The activated sludge process within the pulp and paper industry is generally run to minimize the production of waste activated sludge (WAS), leading to high electricity costs from aeration and relatively large basin volumes. In this study, a pilot-scale activated sludge process was run to evaluate the concept of treating the wastewater at high rate with a low sludge age. Two 150 L containers were used, one for aeration and one for sedimentation and sludge return. The hydraulic retention time was decreased from 24 hours to 7 hours, and the sludge age was lowered from 12 days to 2-4 days. The methane potential of the WAS was evaluated using batch tests, as well as continuous anaerobic digestion (AD) in 4 L reactors in mesophilic and thermophilic conditions. Wastewater treatment capacity was increased almost four-fold at maintained degradation efficiency. The lower sludge age greatly improved the methane potential of the WAS in batch tests, reaching 170 NmL CH 4 /g VS at a sludge age of 2 days. In addition, the continuous AD showed a higher methane production at thermophilic conditions. Thus, the combination of high-rate wastewater treatment and AD of WAS is a promising option for the pulp and paper industry.

  5. Dispersal of thermophilic Desulfotomaculum endospores into Baltic Sea sediments over thousands of years.

    PubMed

    de Rezende, Júlia Rosa; Kjeldsen, Kasper Urup; Hubert, Casey R J; Finster, Kai; Loy, Alexander; Jørgensen, Bo Barker

    2013-01-01

    Patterns of microbial biogeography result from a combination of dispersal, speciation and extinction, yet individual contributions exerted by each of these mechanisms are difficult to isolate and distinguish. The influx of endospores of thermophilic microorganisms to cold marine sediments offers a natural model for investigating passive dispersal in the ocean. We investigated the activity, diversity and abundance of thermophilic endospore-forming sulfate-reducing bacteria (SRB) in Aarhus Bay by incubating pasteurized sediment between 28 and 85 °C, and by subsequent molecular diversity analyses of 16S rRNA and of the dissimilatory (bi)sulfite reductase (dsrAB) genes within the endospore-forming SRB genus Desulfotomaculum. The thermophilic Desulfotomaculum community in Aarhus Bay sediments consisted of at least 23 species-level 16S rRNA sequence phylotypes. In two cases, pairs of identical 16S rRNA and dsrAB sequences in Arctic surface sediment 3000 km away showed that the same phylotypes are present in both locations. Radiotracer-enhanced most probable number analysis revealed that the abundance of endospores of thermophilic SRB in Aarhus Bay sediment was ca. 10(4) per cm(3) at the surface and decreased exponentially to 10(0) per cm(3) at 6.5 m depth, corresponding to 4500 years of sediment age. Thus, a half-life of ca. 300 years was estimated for the thermophilic SRB endospores deposited in Aarhus Bay sediments. These endospores were similarly detected in the overlying water column, indicative of passive dispersal in water masses preceding sedimentation. The sources of these thermophiles remain enigmatic, but at least one source may be common to both Aarhus Bay and Arctic sediments.

  6. Dispersal of thermophilic Desulfotomaculum endospores into Baltic Sea sediments over thousands of years

    PubMed Central

    de Rezende, Júlia Rosa; Kjeldsen, Kasper Urup; Hubert, Casey R J; Finster, Kai; Loy, Alexander; Jørgensen, Bo Barker

    2013-01-01

    Patterns of microbial biogeography result from a combination of dispersal, speciation and extinction, yet individual contributions exerted by each of these mechanisms are difficult to isolate and distinguish. The influx of endospores of thermophilic microorganisms to cold marine sediments offers a natural model for investigating passive dispersal in the ocean. We investigated the activity, diversity and abundance of thermophilic endospore-forming sulfate-reducing bacteria (SRB) in Aarhus Bay by incubating pasteurized sediment between 28 and 85 °C, and by subsequent molecular diversity analyses of 16S rRNA and of the dissimilatory (bi)sulfite reductase (dsrAB) genes within the endospore-forming SRB genus Desulfotomaculum. The thermophilic Desulfotomaculum community in Aarhus Bay sediments consisted of at least 23 species-level 16S rRNA sequence phylotypes. In two cases, pairs of identical 16S rRNA and dsrAB sequences in Arctic surface sediment 3000 km away showed that the same phylotypes are present in both locations. Radiotracer-enhanced most probable number analysis revealed that the abundance of endospores of thermophilic SRB in Aarhus Bay sediment was ca. 104 per cm3 at the surface and decreased exponentially to 100 per cm3 at 6.5 m depth, corresponding to 4500 years of sediment age. Thus, a half-life of ca. 300 years was estimated for the thermophilic SRB endospores deposited in Aarhus Bay sediments. These endospores were similarly detected in the overlying water column, indicative of passive dispersal in water masses preceding sedimentation. The sources of these thermophiles remain enigmatic, but at least one source may be common to both Aarhus Bay and Arctic sediments. PMID:22832348

  7. Concomitant production of cellulase and xylanase by thermophilic mould Sporotrichum thermophile in solid state fermentation and their applicability in bread making.

    PubMed

    Bala, Anju; Singh, Bijender

    2017-06-01

    Sporotrichum thermophile BJAMDU5 secreted high titres of xylanolytic and cellulolytic enzymes in solid state fermentation using mixture of wheat straw and cotton oil cake (ratio 1:1) at 45 °C, pH 5.0 after 72 h inoculated with 2.9 × 10 7  CFU/mL conidiospores. Supplementation of solid medium with lactose and ammonium sulphate further enhanced the production of hydrolytic enzymes. Among different surfactants studied, Tween 80 enhanced the production of all enzymes [3455 U/g DMR (dry mouldy residue), 879.26 U/g DMR, 976.28 U/g DMR and 35.10 U/g DMR for xylanase, CMCase (Carboxymethylcellulase), FPase (Filter paper activity) and β-glucosidase, respectively] as compared to other surfactants. Recycling of solid substrate reduced the production of all these enzymes after second cycle. End products analysis by TLC showed the ability of hydrolytic enzymes of S. thermophile to liberate monomeric (xylose and glucose) as well as oligomeric (xylobiose, cellobiose and higher ones) sugars. Supplementation of enzyme resulted in improved nutritional properties of the bread. Formation of oligomeric sugars by xylanase enzyme of S. thermophile BJAMDU5 make it a good candidate in food industry.

  8. Anaerobic treatment for C and S removal in "zero-discharge" paper mills: effects of process design on S removal efficiencies.

    PubMed

    van Lier, J B; Lens, P N; Pol, L W

    2001-01-01

    Stringent environmental laws in Europe and Northern America lead to the development towards closure of the process water streams in pulp and paper mills. Application of a "zero-discharge" process is already a feasible option for the board and packaging paper industry, provided in-line treatment is applied. Concomitant energy conservation inside the mill results in process water temperatures of 50-60 degrees C. Thermophilic anaerobic treatment complemented with appropriate post-treatment is considered as the most cost-effective solution to meet re-use criteria of the process water and to keep its temperature. In the proposed closed-cycle, the anaerobic treatment step removes the largest fraction of the biodegradable COD and eliminates "S" as H2S from the process stream, without the use of additional chemicals. The anaerobic step is regarded as the only possible location to bleed "S" from the process water cycle. In laboratory experiments, the effect of upward liquid velocity (Vupw) and the specific gas loading rate (Vgas) on the S removal capacity of thermophilic anaerobic bio-reactors was investigated. Acidifying, sulphate reducing sludge bed reactors were fed with partly acidified synthetic paper mill wastewater and were operated at 55 degrees C and pH 6. The reactors were operated at organic loading rates up to 50 g COD.l-1.day-1 at COD/SO4(2-) ratios of 10. The effect of Vupw was researched by comparing the performance of a UASB reactor operated at 1.0 m.h-1 and an EGSB reactor, operated at 6.8 m.h-1. The Vupw had a strong effect on the fermentation patterns. In the UASB reactor, acidification yielded H2, acetate and propionate, leading to an accumulation of reducing equivalents. These were partly disposed of by the production of n-butyrate and n-valerate from propionate. In the EGSB reactor net acetate consumption was observed as well as high volumetric gas (CO2 and CH4) production rates. The higher gas production rates in the EGSB reactor resulted in higher S

  9. Comparison of thermophilic anaerobic digestion characteristics between single-phase and two-phase systems for kitchen garbage treatment.

    PubMed

    Park, YongJin; Hong, Feng; Cheon, JiHoon; Hidaka, Taira; Tsuno, Hiroshi

    2008-01-01

    Lab-scale single-phase and two-phase thermophilic methane fermentation systems (SPS and TPS, respectively) were operated and fed with artificial kitchen waste. In both SPS and TPS, the highest methane recovery ratio of 90%, in terms of chemical oxygen demand by dichromate (CODcr), was observed at an organic loading rate (OLR) of 15 gCODcr/(l.d). The ratio of particle CODcr remaining to total CODcr in the influent was 0.1 and the ratio of NH(4)-N concentration to the input total nitrogen concentration was 0.5 in both SPS and TPS. However, the propionate concentration in the SPS reactor fluctuated largely and was 2 gCODcr/l higher than that in TPS, indicating less stable digestion. Regardless, efficient kitchen waste degradation can be accomplished in both SPS and TPS at an OLR of <20 gCODcr/(l.d), even though TPS may be more stable and easier to maintain. Bacillus coagulans predominated with an occupied ratio of approximately 90% in the acid fermentation reactor of TPS, and then a richer microbial community with a higher Shannon index value was maintained in the methane fermentation reactor of TPS than in the SPS reactor.

  10. Towards a sustainable paradigm of waste-to-energy process: Enhanced anaerobic digestion of sludge with woody biochar

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Shen, Yanwen; Linville, Jessica L.; Ignacio-de Leon, Patricia Anne A.

    This study presents an integrated waste-to-energy process, using two waste streams, sludge generated from the municipal wastewater treatment plants (WWTPs) and biochar generated from the biomass gasification systems, to produce fungible biomethane and nutrient-rich digestate with fertilizer value. Two woody biochar, namely pinewood (PBC) and white oak biochar (WOBC) were used as additives during anaerobic digestion (AD) of WWTP sludge to enhance methane production at mesophilic and thermophilic temperatures. The PBC and WOBC have porous structure, large surface area and desirable chemical properties to be used as AD amendment material to sequester CO2 from biogas in the digester. The biochar-amendedmore » digesters achieved average methane content in biogas of up to 92.3% and 79.0%, corresponding to CO2 sequestration by up to 66.2% and 32.4% during mesophilic and thermophilic AD, respectively. Biochar addition enhanced process stability by increasing the alkalinity, but inhibitory effects were observed at high dosage. It also alleviated free ammonia inhibition by up to 10.5%. The biochar-amended digesters generated digestate rich in macro- and micronutrients including K (up to 300 m/L), Ca (up to 750 mg/L), Mg (up to 1800 mg/L) and Fe (up to 390 mg/L), making biochar-amended digestate a potential alternative used as agricultural lime fertilizer.« less

  11. Genome sequence of the moderately thermophilic sulfur-reducing bacterium Thermanaerovibrio velox type strain (Z-9701 T) and emended description of the genus Thermanaerovibrio

    DOE PAGES

    Palaniappan, Krishna; Meier-Kolthoff, Jan P.; Teshima, Hazuki; ...

    2013-10-16

    Thermanaerovibrio velox Zavarzina et al. 2000 is a member of the Synergistaceae, a family in the phylum Synergistetes that is already well-characterized at the genome level. Members of this phylum were described as Gram-negative staining anaerobic bacteria with a rod/vibrioid cell shape and possessing an atypical outer cell envelope. They inhabit a large variety of anaerobic environments including soil, oil wells, wastewater treatment plants and animal gastrointestinal tracts. They are also found to be linked to sites of human diseases such as cysts, abscesses, and areas of periodontal disease. The moderately thermophilic and organotrophic T. velox shares most of itsmore » morphologic and physiologic features with the closely related species, T. acidaminovorans. In addition to Su883 T, the type strain of T. acidaminovorans, stain Z-9701 T is the second type strain in the genus Thermanaerovibrio to have its genome sequence published. Here we describe the features of this organism, together with the non-contiguous genome sequence and annotation. The 1,880,838 bp long chromosome (non-contiguous finished sequence) with its 1,751 protein-coding and 59 RNA genes is a part of the Genomic Encyclopedia of Bacteria and Archaea project.« less

  12. Genome sequence of the moderately thermophilic sulfur-reducing bacterium Thermanaerovibrio velox type strain (Z-9701 T) and emended description of the genus Thermanaerovibrio

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Palaniappan, Krishna; Meier-Kolthoff, Jan P.; Teshima, Hazuki

    Thermanaerovibrio velox Zavarzina et al. 2000 is a member of the Synergistaceae, a family in the phylum Synergistetes that is already well-characterized at the genome level. Members of this phylum were described as Gram-negative staining anaerobic bacteria with a rod/vibrioid cell shape and possessing an atypical outer cell envelope. They inhabit a large variety of anaerobic environments including soil, oil wells, wastewater treatment plants and animal gastrointestinal tracts. They are also found to be linked to sites of human diseases such as cysts, abscesses, and areas of periodontal disease. The moderately thermophilic and organotrophic T. velox shares most of itsmore » morphologic and physiologic features with the closely related species, T. acidaminovorans. In addition to Su883 T, the type strain of T. acidaminovorans, stain Z-9701 T is the second type strain in the genus Thermanaerovibrio to have its genome sequence published. Here we describe the features of this organism, together with the non-contiguous genome sequence and annotation. The 1,880,838 bp long chromosome (non-contiguous finished sequence) with its 1,751 protein-coding and 59 RNA genes is a part of the Genomic Encyclopedia of Bacteria and Archaea project.« less

  13. Ignavibacterium album gen. nov., sp. nov., a moderately thermophilic anaerobic bacterium isolated from microbial mats at a terrestrial hot spring and proposal of Ignavibacteria classis nov., for a novel lineage at the periphery of green sulfur bacteria.

    PubMed

    Iino, Takao; Mori, Koji; Uchino, Yoshihito; Nakagawa, Tatsunori; Harayama, Shigeaki; Suzuki, Ken-Ichiro

    2010-06-01

    A moderately thermophilic chemoheterotrophic bacterium, strain Mat9-16(T), was isolated from microbial mats developed in hot spring water streams from Yumata, Nagano, Japan. Cells of strain Mat9-16(T) were strictly anaerobic, Gram-stain-negative, non-sporulating, non-motile and short to long rods (2.0-15.5 mum in length). Strain Mat9-16(T) grew fermentatively with optimum growth at 45 degrees C, pH 7.0-7.5 and 1 % NaCl (w/v). Phylogenetic analysis based on the 16S rRNA gene revealed that strain Mat9-16(T) was affiliated with an uncultivated lineage, and the nearest cultivated neighbours were green sulfur bacteria belonging to the class Chlorobea with 77-83 % sequence similarity. However, strain Mat9-16(T) could not grow phototrophically and did not possess light-harvesting structures, morphologically and genetically, such as the chlorosomes of green sulfur bacteria. On the basis of phenotypic features and phylogenetic position, a novel genus and species are proposed for strain Mat9-16(T), to be named Ignavibacterium album gen. nov., sp. nov. (=NBRC 101810(T) =DSM 19864(T)). We also propose to place the cultivated bacterial lineage accommodating the sole representative Mat9-16(T) in a novel class, Ignavibacteria classis nov. In addition, we present a formal description of the phylum-level taxon 'Chlorobi' as Chlorobi phyl. nov.

  14. Aspergillus fumigatus and other thermophilic fungi in nests of wetland birds.

    PubMed

    Korniłłowicz-Kowalska, Teresa; Kitowski, Ignacy

    2013-02-01

    A study was performed on the numbers and species diversity of thermophilic fungi (growing at 45 °C in vitro) in 38 nests of 9 species of wetland birds, taking into account the physicochemical properties of the nests and the bird species. It was found that in nests with the maximum weight (nests of Mute Swan), the number and diversity of thermophilic fungi were significantly greater than in other nests, with lower weight. The diversity of the thermophilic biota was positively correlated with the individual mass of bird and with the level of phosphorus in the nests. The dominant species within the mycobiota under study was Aspergillus fumigatus which inhabited 95% of the nests under study, with average frequency of ca. 650 cfu g(-1) of dry mass of the nest material. In a majority of the nests studied (nests of 7 bird species), the share of A. fumigatus exceeded 50% of the total fungi growing at 45 °C. Significantly higher frequencies of the fungal species were characteristic of the nests of small and medium-sized piscivorous species, compared with the other bird species. The number of A. fumigatus increased with increase in the moisture level of the nests, whereas the frequency of occurrence of that opportunistic pathogen, opposite to the general frequency of thermophilic mycobiota, was negatively correlated with the level of phosphorus in the nest material, and with the body mass and length of the birds. The authors indicate the causes of varied growth of thermophilic fungi in nests of wetland birds and, in particular, present a discussion of the causes of accumulation of A. fumigatus, the related threats to the birds, and its role as a source of transmission in the epidemiological chain of aspergillosis.

  15. Bacillus sp. JR3 esterase LipJ: A new mesophilic enzyme showing traces of a thermophilic past.

    PubMed

    Ribera, Judit; Estupiñán, Mónica; Fuentes, Alba; Fillat, Amanda; Martínez, Josefina; Diaz, Pilar

    2017-01-01

    A search for extremophile enzymes from ancient volcanic soils in El Hierro Island (Canary Islands, Spain) allowed isolation of a microbial sporulated strain collection from which several enzymatic activities were tested. Isolates were obtained after sample cultivation under several conditions of nutrient contents and temperature. Among the bacterial isolates, supernatants from the strain designated JR3 displayed high esterase activity at temperatures ranging from 30 to 100°C, suggesting the presence of at least a hyper-thermophilic extracellular lipase. Sequence alignment of known thermophilic lipases allowed design of degenerated consensus primers for amplification and cloning of the corresponding lipase, named LipJ. However, the cloned enzyme displayed maximum activity at 30°C and pH 7, showing a different profile from that observed in supernatants of the parental strain. Sequence analysis of the cloned protein showed a pentapeptide motif -GHSMG- distinct from that of thermophilic lipases, and much closer to that of esterases. Nevertheless, the 3D structural model of LipJ displayed the same folding as that of thermophilic lipases, suggesting a common evolutionary origin. A phylogenetic study confirmed this possibility, positioning LipJ as a new member of the thermophilic family of bacterial lipases I.5. However, LipJ clusters in a clade close but separated from that of Geobacillus sp. thermophilic lipases. Comprehensive analysis of the cloned enzyme suggests a common origin of LipJ and other bacterial thermophilic lipases, and highlights the most probable divergent evolutionary pathway followed by LipJ, which during the harsh past times would have probably been a thermophilic enzyme, having lost these properties when the environment changed to more benign conditions.

  16. Bacillus sp. JR3 esterase LipJ: A new mesophilic enzyme showing traces of a thermophilic past

    PubMed Central

    Ribera, Judit; Estupiñán, Mónica; Fuentes, Alba; Fillat, Amanda; Martínez, Josefina

    2017-01-01

    A search for extremophile enzymes from ancient volcanic soils in El Hierro Island (Canary Islands, Spain) allowed isolation of a microbial sporulated strain collection from which several enzymatic activities were tested. Isolates were obtained after sample cultivation under several conditions of nutrient contents and temperature. Among the bacterial isolates, supernatants from the strain designated JR3 displayed high esterase activity at temperatures ranging from 30 to 100°C, suggesting the presence of at least a hyper-thermophilic extracellular lipase. Sequence alignment of known thermophilic lipases allowed design of degenerated consensus primers for amplification and cloning of the corresponding lipase, named LipJ. However, the cloned enzyme displayed maximum activity at 30°C and pH 7, showing a different profile from that observed in supernatants of the parental strain. Sequence analysis of the cloned protein showed a pentapeptide motif -GHSMG- distinct from that of thermophilic lipases, and much closer to that of esterases. Nevertheless, the 3D structural model of LipJ displayed the same folding as that of thermophilic lipases, suggesting a common evolutionary origin. A phylogenetic study confirmed this possibility, positioning LipJ as a new member of the thermophilic family of bacterial lipases I.5. However, LipJ clusters in a clade close but separated from that of Geobacillus sp. thermophilic lipases. Comprehensive analysis of the cloned enzyme suggests a common origin of LipJ and other bacterial thermophilic lipases, and highlights the most probable divergent evolutionary pathway followed by LipJ, which during the harsh past times would have probably been a thermophilic enzyme, having lost these properties when the environment changed to more benign conditions. PMID:28742841

  17. Stable isotope probing of acetate fed anaerobic batch incubations shows a partial resistance of acetoclastic methanogenesis catalyzed by Methanosarcina to sudden increase of ammonia level.

    PubMed

    Hao, Liping; Lü, Fan; Mazéas, Laurent; Desmond-Le Quéméner, Elie; Madigou, Céline; Guenne, Angéline; Shao, Liming; Bouchez, Théodore; He, Pinjing

    2015-02-01

    Ammonia inhibition represents a major operational issue for anaerobic digestion. In order to refine our understanding of the terminal catabolic steps in thermophilic anaerobic digestion under ammonia stress, we studied batch thermophilic acetate fed experiments at low (0.26 g L(-1)) and high (7.00 g L(-1)) Total Ammonia Nitrogen concentrations (TAN). Although methane production started immediately for all incubations and resulted in methane yields close to stoichiometric expectations, a 62-72% decrease of methanogenic rate was observed throughout the incubation at 7.00 g L(-1) of TAN compared to 0.26 g L(-1). Stable Isotope Probing analysis of active microbial communities in (13)C-acetate fed experiments coupled to automated ribosomal intergenic spacer analysis and 16S rDNA pyrotag sequencing confirmed that microbial communities were similar for both TAN conditions. At both TAN levels, the (13)C-labeled bacterial community was mainly affiliated to Clostridia-relatives, with OPB54 bacteria being the most abundant sequence in the heavy DNA 16S rDNA pyrotag library. Sequences closely related to Methanosarcina thermophila were also abundantly retrieved in the heavy DNA fractions, showing that this methanogen was still actively assimilating labeled carbon from acetate at free ammonia nitrogen concentrations up to 916 mg L(-1). Stable isotopic signature analysis of biogas, measured in unlabeled acetate fed experiments that were conducted in parallel, confirmed that acetoclastic methanogenic pathway was dominant at both ammonia concentrations. Our work demonstrates that, besides the syntrophic acetate oxidation pathway, acetoclastic methanogenesis catalyzed by Methanosarcina can also play a major role in methane production at high ammonia levels. Copyright © 2014 Elsevier Ltd. All rights reserved.

  18. Thermophilic biofilter for SO2 removal: performance and microbial characteristics.

    PubMed

    Zhang, Jingying; Li, Lin; Liu, Junxin

    2015-03-01

    A bench-scale thermophilic biofilter was applied to remove SO2 at 60°C in the present study. The SO2 concentration in the inlet stream ranged from 100mg/m(3) to 200mg/m(3). An average SO2 removal efficiency of 93.10% was achieved after developing acclimated organisms that can degrade SO2. The thermophilic biofilter effectively reduced SO2, with a maximum elimination capacity of 50.67g/m(3)/h at a loading rate of 51.44g/m(3)/h. Removal efficiency of the thermophilic biofilter was largely influenced by the water containing rate of the packing materials. The SO2 transfer in the biofilter included adsorption by the packing materials, dissolution in liquid, and microbial degradation. The main product of SO2 degradation was SO4(2-). The temporal shifts in the bacterial community that formed in the biofilter were determined through polymerase chain reaction-denaturing gradient gel electrophoresis and DNA sequence analysis. These shifts revealed a correlation between biofilter performance and bacterial community structure. Copyright © 2014 Elsevier Ltd. All rights reserved.

  19. Genetic Tools and Techniques for Recombinant Expression in Thermophilic Bacillaceae.

    PubMed

    Drejer, Eivind B; Hakvåg, Sigrid; Irla, Marta; Brautaset, Trygve

    2018-05-10

    Although Escherichia coli and Bacillus subtilis are the most prominent bacterial hosts for recombinant protein production by far, additional species are being explored as alternatives for production of difficult-to-express proteins. In particular, for thermostable proteins, there is a need for hosts able to properly synthesize, fold, and excrete these in high yields, and thermophilic Bacillaceae represent one potentially interesting group of microorganisms for such purposes. A number of thermophilic Bacillaceae including B. methanolicus , B. coagulans , B. smithii , B. licheniformis , Geobacillus thermoglucosidasius , G. kaustophilus , and G. stearothermophilus are investigated concerning physiology, genomics, genetic tools, and technologies, altogether paving the way for their utilization as hosts for recombinant production of thermostable and other difficult-to-express proteins. Moreover, recent successful deployments of CRISPR/Cas9 in several of these species have accelerated the progress in their metabolic engineering, which should increase their attractiveness for future industrial-scale production of proteins. This review describes the biology of thermophilic Bacillaceae and in particular focuses on genetic tools and methods enabling use of these organisms as hosts for recombinant protein production.

  20. A novel membrane distillation-thermophilic bioreactor system: biological stability and trace organic compound removal.

    PubMed

    Wijekoon, Kaushalya C; Hai, Faisal I; Kang, Jinguo; Price, William E; Guo, Wenshan; Ngo, Hao H; Cath, Tzahi Y; Nghiem, Long D

    2014-05-01

    The removal of trace organic compounds (TrOCs) by a novel membrane distillation-thermophilic bioreactor (MDBR) system was examined. Salinity build-up and the thermophilic conditions to some extent adversely impacted the performance of the bioreactor, particularly the removal of total nitrogen and recalcitrant TrOCs. While most TrOCs were well removed by the thermophilic bioreactor, compounds containing electron withdrawing functional groups in their molecular structure were recalcitrant to biological treatment and their removal efficiency by the thermophilic bioreactor was low (0-53%). However, the overall performance of the novel MDBR system with respect to the removal of total organic carbon, total nitrogen, and TrOCs was high and was not significantly affected by the conditions of the bioreactor. All TrOCs investigated here were highly removed (>95%) by the MDBR system. Biodegradation, sludge adsorption, and rejection by MD contribute to the removal of TrOCs by MDBR treatment. Crown Copyright © 2014. Published by Elsevier Ltd. All rights reserved.

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

    PubMed

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

    2004-02-01

    The diversity, population dynamics, and activity profiles of methanogens in anaerobic granular sludges from two anaerobic hybrid reactors treating a molasses wastewater both mesophilically (37 degrees C) and thermophilically (55 degrees C) during a 1081 day trial were determined. The influent to one of the reactors was supplemented with sulphate, after an acclimation period of 112 days, to determine the effect of competition with sulphate-reducing bacteria on the methanogenic community structure. Sludge samples were removed from the reactors at intervals throughout the operational period and examined by amplified ribosomal DNA (rDNA) restriction analysis (ARDRA) and partial sequencing of 16S rRNA genes. In total, 18 operational taxonomic units (OTUs) were identified, 12 of which were sequenced. The methanogenic communities in both reactors changed during the operational period. The seed sludge and the reactor biomass sampled during mesophilic operation, both in the presence and absence of sulphate, was characterised by a predominance of Methanosaeta spp. Following temperature elevation, the dominant methanogenic sequences detected in the non-sulphate supplemented reactor were closely related to Methanocorpusculum parvum. By contrast, the dominant OTUs detected in the sulphate-supplemented reactor upon temperature increase were related to the hydrogen-utilising methanogen, Methanobacterium thermoautotrophicum. The observed methanogenic community structure in the reactors correlated with the operational performance of the reactors during the trial and with physiological measurements of the reactor biomass. Both reactors achieved chemical oxygen demand (COD) removal efficiencies of over 90% during mesophilic operation, with or without sulphate supplementation. During thermophilic operation, the presence of sulphate resulted in decreased reactor performance (effluent acetate concentrations of >3000 mg/l and biogas methane content of <25%). It was demonstrated that

  2. Methane yields and methanogenic community changes during co-fermentation of cattle slurry with empty fruit bunches of oil palm.

    PubMed

    Walter, Andreas; Franke-Whittle, Ingrid H; Wagner, Andreas O; Insam, Heribert

    2015-01-01

    The biomethane potential and structural changes of the methanogenic community in a solid-state anaerobic digestion process co-digesting cattle slurry and empty fruit bunches were investigated under mesophilic (37°C) and thermophilic (55°C) conditions. Phylogenetic microarrays revealed the presence of two hydrogenotrophic genera (Methanoculleus and Methanobrevibacter) and one acetoclastic genus (Methanosarcina). Methanosarcina numbers were found to increase in both mesophilic and thermophilic treatments of empty fruit bunches. Methanobrevibacter, which dominated in the cattle slurry, remained constant during anaerobic digestion (AD) at 37°C and decreased in numbers during digestion at 55°C. Numbers of Methanoculleus remained constant at 37°C and increased during the thermophilic digestion. Physicochemical data revealed non-critical concentrations for important monitoring parameters such as total ammonia nitrogen, free ammonia nitrogen and volatile fatty acids in all treatments after AD. The biomethane potential of empty fruit bunches was higher under thermophilic conditions than under mesophilic conditions. Copyright © 2014 Elsevier Ltd. All rights reserved.

  3. Mesophilic-hydrothermal-thermophilic (M-H-T) digestion of green corn straw.

    PubMed

    Li, Dong; Wang, Qingjing; Li, Jiang; Li, Zhidong; Yuan, Yuexiang; Yan, Zhiying; Mei, Zili; Liu, Xiaofeng

    2016-02-01

    Mesophilic-hydrothermal (80-160 °C, 30 min)-thermophilic (M-H-T) digestion and control tests of mesophilic (M), thermophilic (T), hydrothermal-mesophilic (H-M), and mesophilic-thermophilic digestion (M-T) of green corn straw were conducted for a 20-day fermentation period. The results indicate that M-H-T is an efficient method to improve methane production. A maximum methane yield of 371.74 mL/g volatile solid was obtained by the M (3 days)-H (140 °C)-T (17 days) process, which was 20.44%, 16.55%, 31.44%, and 14.31% higher than the yields of the M, T, 140-M, and M-T processes. The enhanced methane production was attributed to (1) the improved hemicellulose degradation and lignin disorganization; (2) prevention of the degradation of soluble sugar, easily hydrolyzed hemicellulose and cellulose into furfural and methylfurfural; and (3) lack of formation of Maillard reaction products during initial hydrothermal treatment. Copyright © 2015 Elsevier Ltd. All rights reserved.

  4. Thermophilic bacteria in Moroccan hot springs, salt marshes and desert soils.

    PubMed

    Aanniz, Tarik; Ouadghiri, Mouna; Melloul, Marouane; Swings, Jean; Elfahime, Elmostafa; Ibijbijen, Jamal; Ismaili, Mohamed; Amar, Mohamed

    2015-06-01

    The diversity of thermophilic bacteria was investigated in four hot springs, three salt marshes and 12 desert sites in Morocco. Two hundred and forty (240) thermophilic bacteria were recovered, identified and characterized. All isolates were Gram positive, rod-shaped, spore forming and halotolerant. Based on BOXA1R-PCR and 16S rRNA gene sequencing, the recovered isolates were dominated by the genus Bacillus (97.5%) represented by B. licheniformis (119), B. aerius (44), B. sonorensis (33), B. subtilis (subsp. spizizenii (2) and subsp. inaquosurum (6)), B. amyloliquefaciens (subsp. amyloliquefaciens (4) and subsp. plantarum (4)), B. tequilensis (3), B. pumilus (3) and Bacillus sp. (19). Only six isolates (2.5%) belonged to the genus Aeribacillus represented by A. pallidus (4) and Aeribacillus sp. (2). In this study, B. aerius and B. tequilensis are described for the first time as thermophilic bacteria. Moreover, 71.25%, 50.41% and 5.41% of total strains exhibited high amylolytic, proteolytic or cellulolytic activity respectively.

  5. Thermophilic bacteria in Moroccan hot springs, salt marshes and desert soils

    PubMed Central

    Aanniz, Tarik; Ouadghiri, Mouna; Melloul, Marouane; Swings, Jean; Elfahime, Elmostafa; Ibijbijen, Jamal; Ismaili, Mohamed; Amar, Mohamed

    2015-01-01

    The diversity of thermophilic bacteria was investigated in four hot springs, three salt marshes and 12 desert sites in Morocco. Two hundred and forty (240) thermophilic bacteria were recovered, identified and characterized. All isolates were Gram positive, rod-shaped, spore forming and halotolerant. Based on BOXA1R-PCR and 16S rRNA gene sequencing, the recovered isolates were dominated by the genus Bacillus (97.5%) represented by B. licheniformis (119), B. aerius (44), B. sonorensis (33), B. subtilis (subsp. spizizenii (2) and subsp. inaquosurum (6)), B. amyloliquefaciens (subsp. amyloliquefaciens (4) and subsp. plantarum (4)), B. tequilensis (3), B. pumilus (3) and Bacillus sp. (19). Only six isolates (2.5%) belonged to the genus Aeribacillus represented by A. pallidus (4) and Aeribacillus sp. (2). In this study, B. aerius and B. tequilensis are described for the first time as thermophilic bacteria. Moreover, 71.25%, 50.41% and 5.41% of total strains exhibited high amylolytic, proteolytic or cellulolytic activity respectively. PMID:26273259

  6. New Strategy for a Suitable Fast Stabilization of the Biomethanization Performance

    PubMed Central

    Fernández-Güelfo, L. A.; Álvarez-Gallego, C. J.; Sales Márquez, D.; Romero García, L. I.

    2012-01-01

    The start-up strategies for thermophilic anaerobic reactors usually consist of an initial mesophilic stage (35°C), with an approximate duration of 185 days, and a subsequent thermophilic stage (55°C), which normally requires around 60 days to achieve the system stabilizatio. During the first 8–10 days of the mesophilic stage, the reactor is not fed so that the inoculum, which is generally a mesophilic anaerobic sludge, may be adapted to the organic solid waste. Between mesophilic and thermophilic conditions the reactor is still not fed in an effort to prevent possible imbalances in the proces. As a consequence, the start-up and stabilization of the biomethanization performance described in the literature require, at least, around 245 days. In this sense, a new strategy for the start-up and stabilization phases is presented in this study. This approach allows an important reduction in the overall time necessary for these stages in an anaerobic continuous stirred tank reactor (CSTR) operated at thermophilic-dry conditions for treating the organic fraction of the municipal solid waste (OFMSW): 60 days versus 245 days of conventional strategies. The new strategy uses modified SEBAC technology to adapt an inoculum to the OFMSW and the operational conditions prior to seeding the CSTR. PMID:23193374

  7. Effects of influent fractionation, kinetics, stoichiometry and mass transfer on CH4, H2 and CO2 production for (plant-wide) modeling of anaerobic digesters.

    PubMed

    Solon, Kimberly; Flores-Alsina, Xavier; Gernaey, Krist V; Jeppsson, Ulf

    2015-01-01

    This paper examines the importance of influent fractionation, kinetic, stoichiometric and mass transfer parameter uncertainties when modeling biogas production in wastewater treatment plants. The anaerobic digestion model no. 1 implemented in the plant-wide context provided by the benchmark simulation model no. 2 is used to quantify the generation of CH₄, H₂and CO₂. A comprehensive global sensitivity analysis based on (i) standardized regression coefficients (SRC) and (ii) Morris' screening's (MS's) elementary effects reveals the set of parameters that influence the biogas production uncertainty the most. This analysis is repeated for (i) different temperature regimes and (ii) different solids retention times (SRTs) in the anaerobic digester. Results show that both SRC and MS are good measures of sensitivity unless the anaerobic digester is operating at low SRT and mesophilic conditions. In the latter situation, and due to the intrinsic nonlinearities of the system, SRC fails in decomposing the variance of the model predictions (R² < 0.7) making MS a more reliable method. At high SRT, influent fractionations are the most influential parameters for predictions of CH₄and CO₂emissions. Nevertheless, when the anaerobic digester volume is decreased (for the same load), the role of acetate degraders gains more importance under mesophilic conditions, while lipids and fatty acid metabolism is more influential under thermophilic conditions. The paper ends with a critical discussion of the results and their implications during model calibration and validation exercises.

  8. Growth of the facultative anaerobe Shewanella putrefaciens by elemental sulfur reduction

    NASA Technical Reports Server (NTRS)

    Moser, D. P.; Nealson, K. H.

    1996-01-01

    The growth of bacteria by dissimilatory elemental sulfur reduction is generally associated with obligate anaerobes and thermophiles in particular. Here we describe the sulfur-dependent growth of the facultatively anaerobic mesophile Shewanella putrefaciens. Six of nine representative S. putrefaciens isolates from a variety of environments proved able to grow by sulfur reduction, and strain MR-1 was chosen for further study. Growth was monitored in a minimal medium (usually with 0.05% Casamino Acids added as a growth stimulant) containing 30 mM lactate and limiting concentrations of elemental sulfur. When mechanisms were provided for the removal of the metabolic end product, H2S, measurable growth was obtained at sulfur concentrations of from 2 to 30 mM. Initial doubling times were ca. 1.5 h and substrate independent over the range of sulfur concentrations tested. In the cultures with the highest sulfur concentrations, cell numbers increased by greater than 400-fold after 48 h, reaching a maximum density of 6.8 x 10(8) cells ml-1. Yields were determined as total cell carbon and ranged from 1.7 to 5.9 g of C mol of S(0) consumed-1 in the presence of the amino acid supplement and from 0.9 to 3.4 g of C mol of S(0-1) in its absence. Several lines of evidence indicate that cell-to-sulfur contact is not required for growth. Approaches for the culture of sulfur-metabolizing bacteria and potential ecological implications of sulfur reduction in Shewanella-like heterotrophs are discussed.

  9. Effect of temperature on solids reductions and on degradation kinetics during thermophilic aerobic digestion of a simulated sludge.

    PubMed

    Toki, C J

    2008-07-01

    Laboratory-scale experiments were conducted to determine the influence of higher thermophilic temperatures on thermophilic aerobic digestion treatment of a simulated sludge. The efficiency of the process was evaluated in respect of solids removal and degradation rate constants at four thermophilic temperatures. Batch runs were operated at a retention time of one day and temperatures of 65, 70, 72 and 75 degrees C. The results indicated that temperature increase did not impart any significant benefits to the digestion operation in terms of suspended solids and biochemichal oxygen demand reduction. The findings from this research also suggested that the treatment would not appear to benefit from temperatures higher than 65 degrees C, as classically suggested by Van't Hoff-Arrhenius. Therefore, increase of thermophilic temperature in the tested 65-75 degrees C range does not enhance the efficiency of thermophilic, aerobic sludge digestion treatment.

  10. Potential for energy generation from anaerobic digestion of food waste in Australia.

    PubMed

    Lou, Xian Fang; Nair, Jaya; Ho, Goen

    2013-03-01

    Published national and state reports have revealed that Australia deposits an average of 16 million Mg of solid waste into landfills yearly, of which approximately 12.6% is comprised of food. Being highly biodegradable and possessing high energy content, anaerobic digestion offers an attractive treatment option alternative to landfilling. The present study attempted to identify the theoretical maximum benefit of food waste digestion in Australia with regard to energy recovery and waste diversion from landfills. The study also assessed the scope for anaerobic process to utilize waste for energy projects through various case study scenarios. Results indicated anaerobic digestion of total food waste generated across multiple sites in Australia could generate 558 453 dam(3) of methane which translated to 20.3 PJ of heating potential or 1915 GWe in electricity generation annually. This would contribute to 3.5% of total current energy supply from renewable sources. Energy contribution from anaerobic digestion of food waste to the total energy requirement in Australia remains low, partially due to the high energy consumption of the country. However its appropriateness in low density regions, which are prevalent in Australia, may allow digesters to have a niche application in the country.

  11. Indigenous microbial capability in solid manure residues to start-up solid-phase anaerobic digesters.

    PubMed

    Yap, S D; Astals, S; Jensen, P D; Batstone, D J; Tait, S

    2017-06-01

    Batch solid-phase anaerobic digestion is a technology for sustainable on-farm treatment of solid residues, but is an emerging technology that is yet to be optimised with respect to start-up and inoculation. In the present study, spent bedding from two piggeries (site A and B) were batch digested at total solids (TS) concentration of 5, 10 and 20% at mesophilic (37°C) and thermophilic (55°C) temperatures, without adding an external inoculum. The results showed that the indigenous microbial community present in spent bedding was able to recover the full methane potential of the bedding (140±5 and 227±6L CH 4 kgVS fed -1 for site A and B, respectively), but longer treatment times were required than for digestion with an added external inoculum. Nonetheless, at high solid loadings (i.e. TS level>10%), the digestion performance was affected by chemical inhibition due to ammonia and/or humic acid. Thermophilic temperatures did not influence digestion performance but did increase start-up failure risk. Further, inoculation of residues from the batch digestion to subsequent batch enhanced start-up and achieved full methane potential recovery of the bedding. Inoculation with liquid residue (leachate) was preferred over a solid residue, to preserve treatment capacity for fresh substrate. Overall, the study highlighted that indigenous microbial community in the solid manure residue was capable of recovering full methane potential and that solid-phase digestion was ultimately limited by chemical inhibition rather than lack of suitable microbial community. Copyright © 2017 Elsevier Ltd. All rights reserved.

  12. Characterization of thermophilic fungal community associated with pile fermentation of Pu-erh tea.

    PubMed

    Zhang, Wei; Yang, Ruijuan; Fang, Wenjun; Yan, Liang; Lu, Jun; Sheng, Jun; Lv, Jie

    2016-06-16

    This study aimed to characterize the thermophilic fungi in pile-fermentation process of Pu-erh tea. Physicochemical analyses showed that the high temperature and low pH provided optimal conditions for propagation of fungi. A number of fungi, including Blastobotrys adeninivorans, Thermomyces lanuginosus, Rasamsonia emersonii, Aspergillus fumigatus, Rhizomucor pusillus, Rasamsonia byssochlamydoides, Rasamsonia cylindrospora, Aspergillus tubingensis, Aspergillus niger, Candida tropicalis and Fusarium graminearum were isolated as thermophilic fungi under combination of high temperature and acid culture conditions from Pu-erh tea pile-fermentation. The fungal communities were analyzed by PCR-DGGE. Results revealed that those fungi are closely related to Debaryomyces hansenii, Cladosporium cladosporioides, A. tubingensis, R. emersonii, R. pusillus, A. fumigatus and A. niger, and the last four presented as dominant species in the pile process. These four preponderant thermophilic fungi reached the order of magnitude of 10(7), 10(7), 10(7) and 10(6)copies/g dry tea, respectively, measured by real-time quantitative PCR (q-PCR). The results indicate that the thermophilic fungi play an important role in Pu-erh tea pile fermentation. Copyright © 2016 Elsevier B.V. All rights reserved.

  13. Conductive heating and microwave hydrolysis under identical heating profiles for advanced anaerobic digestion of municipal sludge.

    PubMed

    Mehdizadeh, Seyedeh Neda; Eskicioglu, Cigdem; Bobowski, Jake; Johnson, Thomas

    2013-09-15

    Microwave (2.45 GHz, 1200 W) and conventional heating (custom pressure vessel) pretreatments were applied to dewatered municipal waste sludge (18% total solids) using identical heating profiles that span a wide range of temperatures (80-160 °C). Fourteen lab-scale semi-continuous digesters were set up to optimize the energy (methane) output and sludge retention time (SRT) requirements of untreated (control) and thermally pretreated anaerobic digesters operated under mesophilic and thermophilic temperatures. Both pretreatment methods indicated that in the pretreatment range of 80-160 °C, temperature was a statistically significant factor (p-value < 0.05) for increasing solubilization of chemical oxygen demand and biopolymers (proteins, sugars, humic acids) of the waste sludge. However, the type of pretreatment method, i.e. microwave versus conventional heating, had no statistically significant effect (p-value >0.05) on sludge solubilization. With the exception of the control digesters at a 5-d SRT, all control and pretreated digesters achieved steady state at all three SRTs, corresponding to volumetric organic loading rates of 1.74-6.96 g chemical oxygen demand/L/d. At an SRT of 5 d, both mesophilic and thermophilic controls stopped producing biogas after 20 d of operation with total volatile fatty acids concentrations exceeding 1818 mg/L at pH <5.64 for mesophilic and 2853 mg/L at pH <7.02 for thermophilic controls, while the pretreated digesters continued producing biogas. Furthermore, relative (to control) organic removal efficiencies dramatically increased as SRT was shortened from 20 to 10 and then 5 d, indicating that the control digesters were challenged as the organic loading rate was increased. Energy analysis showed that, at an elevated temperature of 160 °C, the amount of methane recovered was not enough to compensate for the energy input. Among the digesters with positive net energy productions, control and pretreated digesters at 80 °C were more

  14. Novel thermophilic hemicellulases for the conversion of lignocellulose for second generation biorefineries.

    PubMed

    Cobucci-Ponzano, Beatrice; Strazzulli, Andrea; Iacono, Roberta; Masturzo, Giuseppe; Giglio, Rosa; Rossi, Mosè; Moracci, Marco

    2015-10-01

    The biotransformation of lignocellulose biomasses into fermentable sugars is a very complex procedure including, as one of the most critical steps, the (hemi) cellulose hydrolysis by specific enzymatic cocktails. We explored here, the potential of stable glycoside hydrolases from thermophilic organisms, so far not used in commercial enzymatic preparations, for the conversion of glucuronoxylan, the major hemicellulose of several energy crops. Searches in the genomes of thermophilic bacteria led to the identification, efficient production, and detailed characterization of novel xylanase and α-glucuronidase from Alicyclobacillus acidocaldarius (GH10-XA and GH67-GA, respectively) and a α-glucuronidase from Caldicellulosiruptor saccharolyticus (GH67-GC). Remarkably, GH10-XA, if compared to other thermophilic xylanases from this family, coupled good specificity on beechwood xylan and the best stability at 65 °C (3.5 days). In addition, GH67-GC was the most stable α-glucuronidases from this family and the first able to hydrolyse both aldouronic acid and aryl-α-glucuronic acid substrates. These enzymes, led to the very efficient hydrolysis of beechwood xylan by using 7- to 9-fold less protein (concentrations <0.3 μM) and in much less reaction time (2h vs 12h) if compared to other known biotransformations catalyzed by thermophilic enzymes. In addition, remarkably, together with a thermophilic β-xylosidase, they catalyzed the production of xylose from the smart cooking pre-treated biomass of one of the most promising energy crops for second generation biorefineries. We demonstrated that search by the CAZy Data Bank of currently available genomes and detailed enzymatic characterization of recombinant enzymes allow the identification of glycoside hydrolases with novel and interesting properties and applications. Copyright © 2015 Elsevier Inc. All rights reserved.

  15. Structural adaptation of the subunit interface of oligomeric thermophilic and hyperthermophilic enzymes.

    PubMed

    Maugini, Elisa; Tronelli, Daniele; Bossa, Francesco; Pascarella, Stefano

    2009-04-01

    Enzymes from thermophilic and, particularly, from hyperthermophilic organisms are surprisingly stable. Understanding of the molecular origin of protein thermostability and thermoactivity attracted the interest of many scientist both for the perspective comprehension of the principles of protein structure and for the possible biotechnological applications through application of protein engineering. Comparative studies at sequence and structure levels were aimed at detecting significant differences of structural parameters related to protein stability between thermophilic and hyperhermophilic structures and their mesophilic homologs. Comparative studies were useful in the identification of a few recurrent themes which the evolution utilized in different combinations in different protein families. These studies were mostly carried out at the monomer level. However, maintenance of a proper quaternary structure is an essential prerequisite for a functional macromolecule. At the environmental temperatures experienced typically by hyper- and thermophiles, the subunit interactions mediated by the interface must be sufficiently stable. Our analysis was therefore aimed at the identification of the molecular strategies adopted by evolution to enhance interface thermostability of oligomeric enzymes. The variation of several structural properties related to protein stability were tested at the subunit interfaces of thermophilic and hyperthermophilic oligomers. The differences of the interface structural features observed between the hyperthermophilic and thermophilic enzymes were compared with the differences of the same properties calculated from pairwise comparisons of oligomeric mesophilic proteins contained in a reference dataset. The significance of the observed differences of structural properties was measured by a t-test. Ion pairs and hydrogen bonds do not vary significantly while hydrophobic contact area increases specially in hyperthermophilic interfaces. Interface

  16. Microbiology of Kamchatka Peninsula Hot Springs

    NASA Astrophysics Data System (ADS)

    Bonch-Osmolovsk, E.

    2005-12-01

    Hot springs of Uzon Caldera, Geyser Valley, Moutnovsky Volcano (Kamchatka Peninsula) served as the sources of isolation of numerous thermophilic prokaryotes, many of them representing new taxa. Among new isolates there were hyperthermophilic archaea - neutrophilic or acidophilic anaerobic organotrophs, able to use a wide range of polymeric organic substrates. Bacterial isolates were in majority represented by moderate thermophiles - organotrophs and lithoautotrophs. Latter group consisted of anaerobes oxidizing molecular hydrogen in the course of sulfate, sulfur or iron reduction, and of anaerobic CO-oxidizing, hydrogen-producing bacteria. Some of new isolates represented deep phylogenetic lineages in Bacteria domain. Microbial activity in Kamchatka hot springs was studied by means of radioisotopic tracing. The rates of methanogenesis, acetogenesis, inorganic carbon assimilation, acetate oxidation were determined in three different hot springs with pH ranging from 3.0 to 8.5 and water temeperature being in the range from 55 to 85oC. The results indicated the presence and activity of novel metabolic groups of thermophilic prokaryotes that so far have not been known in laboratory cultures.

  17. Effect of temperature and temperature fluctuation on thermophilic anaerobic digestion of cattle manure.

    PubMed

    El-Mashad, Hamed M; Zeeman, Grietje; van Loon, Wilko K P; Bot, Gerard P A; Lettinga, Gatze

    2004-11-01

    The influence of temperature, 50 and 60 degrees C, at hydraulic retention times (HRTs) of 20 and 10 days, on the performance of anaerobic digestion of cow manure has been investigated in completely stirred tank reactors (CSTRs). Furthermore, the effect of both daily downward and daily upward temperature fluctuations has been studied. In the daily downward temperature fluctuation regime the temperatures of each reactor was reduced by 10 degrees C for 10 h while in the daily upward fluctuation regime the temperature of each reactor was increased 10 degrees C for 5 h. The results show that the methane production rate at 60 degrees C is lower than that at 50 degrees C at all experimental conditions of imposed HRT except when downward temperature fluctuations were applied at an HRT of 10 days. It also was found that the free ammonia concentration not only affects the acetate-utilising bacteria but also the hydrolysis and acidification process. The upward temperature fluctuation affects the maximum specific methanogenesis activity more severely as compared to imposed downward temperature fluctuations. The results clearly reveal the possibility of using available solar energy at daytime to heat up the reactor(s) without the need of heat storage during nights, especially at an operational temperature of 50 degrees C and at a 20 days HRT, and without the jeopardising of the overheating.

  18. Integration between chemical oxidation and membrane thermophilic biological process.

    PubMed

    Bertanza, G; Collivignarelli, M C; Crotti, B M; Pedrazzani, R

    2010-01-01

    Full scale applications of activated sludge thermophilic aerobic process for treatment of liquid wastes are rare. This experimental work was carried out at a facility, where a thermophilic reactor (1,000 m(3) volume) is operated. In order to improve the global performance of the plant, it was decided to upgrade it, by means of two membrane filtration units (ultrafiltration -UF-, in place of the final sedimentation, and nanofiltration -NF-). Subsequently, the integration with chemical oxidation (O(3) and H(2)O(2)/UV processes) was taken into consideration. Studied solutions dealt with oxidation of both the NF effluents (permeate and concentrate). Based on experimental results and economic evaluation, an algorithm was proposed for defining limits of convenience of this process.

  19. Microbial characteristics in anaerobic digestion process of food waste for methane production-A review.

    PubMed

    Wang, Pan; Wang, Hongtao; Qiu, Yinquan; Ren, Lianhai; Jiang, Bin

    2018-01-01

    Food waste (FW) is rich in starch, fat, protein and cellulose. It is easy to decay and brings environmental pollution and other social problems. FW shows a high potential to produce methane by anaerobic digestion (AD) due to its high organic content. However, many inhibitors, such as accumulation of ammonia and volatile fatty acids (VFAs), usually result in inefficient performances and even process failure. Microorganisms play an important role in the process of hydrolysis, acidogenesis, acetogenesis and methanogenesis. This review provided a critical summary of microbial characteristics to obtain connects of microbial community structure with operational conditions at various states of AD, such as mesophilic and thermophilic, wet and dry, success and failure, pretreated or not, lab-scale and full-scale. This article emphasizes that it is necessary to analyze changes and mechanisms of microbial communities in unbalanced system and seek efficiency dynamic succession rules of the dominant microorganisms. Copyright © 2017. Published by Elsevier Ltd.

  20. Long-Term Enrichment on Cellulose or Xylan Causes Functional and Taxonomic Convergence of Microbial Communities from Anaerobic Digesters

    PubMed Central

    Jia, Yangyang; Wilkins, David; Lu, Hongyuan; Cai, Mingwei

    2015-01-01

    Cellulose and xylan are two major components of lignocellulosic biomass, which represents a potentially important energy source, as it is abundant and can be converted to methane by microbial action. However, it is recalcitrant to hydrolysis, and the establishment of a complete anaerobic digestion system requires a specific repertoire of microbial functions. In this study, we maintained 2-year enrichment cultures of anaerobic digestion sludge amended with cellulose or xylan to investigate whether a cellulose- or xylan-digesting microbial system could be assembled from sludge previously used to treat neither of them. While efficient methane-producing communities developed under mesophilic (35°C) incubation, they did not under thermophilic (55°C) conditions. Illumina amplicon sequencing results of the archaeal and bacterial 16S rRNA genes revealed that the mature cultures were much lower in richness than the inocula and were dominated by single archaeal (genus Methanobacterium) and bacterial (order Clostridiales) groups, although at finer taxonomic levels the bacteria were differentiated by substrates. Methanogenesis was primarily via the hydrogenotrophic pathway under all conditions, although the identity and growth requirements of syntrophic acetate-oxidizing bacteria were unclear. Incubation conditions (substrate and temperature) had a much greater effect than inoculum source in shaping the mature microbial community, although analysis based on unweighted UniFrac distance found that the inoculum still determined the pool from which microbes could be enriched. Overall, this study confirmed that anaerobic digestion sludge treating nonlignocellulosic material is a potential source of microbial cellulose- and xylan-digesting functions given appropriate enrichment conditions. PMID:26712547

  1. Cleavage of the main carbon chain backbone of high molecular weight polyacrylamide by aerobic and anaerobic biological treatment.

    PubMed

    Song, Wenzhe; Zhang, Yu; Gao, Yingxin; Chen, Dong; Yang, Min

    2017-12-01

    High molecular weight partially hydrolyzed polyacrylamide (PAM) can be bio-hydrolyzed on the amide side group, however, solid evidence regarding the biological cleavage of its main carbon chain backbone is limited. In this study, viscometry, flow field-flow fractionation multi-angle light scattering (FFF-MALS), and attenuated total reflectance-Fourier transform infrared (ATR-FTIR) analysis were used to investigate the biodegradability of PAM with a nominal molecular weight of 2 × 10 7  Da (Da) in two suspended aerobic (25 and 40 °C) and two upflow anaerobic blanket reactors (35 and 55 °C) operated for 470 d under a hydraulic residence time (HRT) of 2 d. Both anaerobic and aerobic biological treatment reduced the viscosity from 2.02 cp in the influent to 1.45-1.60 cp, and reduced the molecular weight of PAM using FFF-MALS from 2.17 × 10 7  Da to less than one-third its original size. The removals of both the amide group and carbon chain backbone in the PAM molecule were further supported by the FTIR analysis. In comparison with the other conditions, thermophilic anaerobic treatment exhibited higher efficiency for PAM biodegradation. Batch test excluded the influence of temperature on the molecular weight of PAM over the range 25-55 °C, suggesting that cleavage of the main carbon chain backbone was attributed to biological degradation. Our results suggested that high molecular weight PAM was biodegradable, but mineralization did not occur. Copyright © 2017 Elsevier Ltd. All rights reserved.

  2. Direct bioconversion of brown algae into ethanol by thermophilic bacterium Defluviitalea phaphyphila.

    PubMed

    Ji, Shi-Qi; Wang, Bing; Lu, Ming; Li, Fu-Li

    2016-01-01

    Brown algae are promising feedstocks for biofuel production with inherent advantages of no structural lignin, high growth rate, and no competition for land and fresh water. However, it is difficult for one microorganism to convert all components of brown algae with different oxidoreduction potentials to ethanol. Defluviitalea phaphyphila Alg1 is the first characterized thermophilic bacterium capable of direct utilization of brown algae. Defluviitalea phaphyphila Alg1 can simultaneously utilize mannitol, glucose, and alginate to produce ethanol, and high ethanol yields of 0.47 g/g-mannitol, 0.44 g/g-glucose, and 0.3 g/g-alginate were obtained. A rational redox balance system under obligate anaerobic condition in fermenting brown algae was revealed in D. phaphyphila Alg1 through genome and redox analysis. The excess reducing equivalents produced from mannitol metabolism were equilibrated by oxidizing forces from alginate assimilation. Furthermore, D. phaphyphila Alg1 can directly utilize unpretreated kelp powder, and 10 g/L of ethanol was accumulated within 72 h with an ethanol yield of 0.25 g/g-kelp. Microscopic observation further demonstrated the deconstruction process of brown algae cell by D. phaphyphila Alg1. The integrated biomass deconstruction system of D. phaphyphila Alg1, as well as its high ethanol yield, provided us an excellent alternative for brown algae bioconversion at elevated temperature.

  3. Thermoanaerobacter uzonensis sp. nov., an anaerobic thermophilic bacterium isolated from a hot spring within the Uzon Caldera, Kamchatka, Far East Russia.

    PubMed

    Wagner, Isaac D; Zhao, Weidong; Zhang, Chuanlun L; Romanek, Christopher S; Rohde, Manfred; Wiegel, Juergen

    2008-11-01

    Several strains of heterotrophic, anaerobic thermophilic bacteria were isolated from hot springs of the Uzon Caldera, Kamchatka, Far East Russia. Strain JW/IW010(T) was isolated from a hot spring within the West sector of the Eastern Thermal field, near Pulsating Spring in the Winding Creek area. Cells of strain JW/IW010(T) were straight to slightly curved rods, 0.5 mum in width and variable in length from 2 to 5 mum and occasionally up to 15 mum, and formed oval subterminal spores. Cells stained Gram-negative, but were Gram-type positive. Growth was observed between 32.5 and 69 degrees C with an optimum around 61 degrees C (no growth occurred at or below 30 degrees C, or at or above 72 degrees C). The pH(60 degrees C) range for growth was 4.2-8.9 with an optimum at 7.1 (no growth occurred at or below pH(60 degrees C) 3.9, or at 9.2 or above). The shortest observed doubling-time at pH(60 degrees C) 6.9 and 61 degrees C was 30 min. Strain JW/IW010(T) was chemo-organotrophic; yeast extract, peptone, Casamino acids and tryptone supported growth. Yeast extract was necessary for the utilization of non-proteinaceous substrates, and growth was observed with inulin, cellobiose, maltose, sucrose, glucose, fructose, galactose, mannose, xylose, trehalose, mannitol, pyruvate and crotonate. The G+C content of the genomic DNA of strain JW/IW010(T) was 33.6 mol% (HPLC method). The major phospholipid fatty acids were iso-15 : 0 (53.5 %), 15 : 0 (11.8 %), 16 : 0 (7.3 %), 10-methyl 16 : 0 (7.3 %) and anteiso-15 : 0 (5.3 %). 16S rRNA gene sequence analysis placed strain JW/IW010(T) in the genus Thermoanaerobacter of the family 'Thermoanaerobacteriaceae' (Firmicutes), with Thermoanaerobacter sulfurigignens JW/SL-NZ826(T) (97 % 16S rRNA gene sequence similarity) and Thermoanaerobacter kivui DSM 2030(T) (94.5 %) as the closest phylogenetic relatives with validly published names. The level of DNA-DNA relatedness between strain JW/IW010(T) and Thermoanaerobacter sulfurigignens JW/SL-NZ826

  4. Extremely thermophilic microorganisms for biomass conversion: status and prospects.

    PubMed

    Blumer-Schuette, Sara E; Kataeva, Irina; Westpheling, Janet; Adams, Michael Ww; Kelly, Robert M

    2008-06-01

    Many microorganisms that grow at elevated temperatures are able to utilize a variety of carbohydrates pertinent to the conversion of lignocellulosic biomass to bioenergy. The range of substrates utilized depends on growth temperature optimum and biotope. Hyperthermophilic marine archaea (T(opt)>or=80 degrees C) utilize alpha- and beta-linked glucans, such as starch, barley glucan, laminarin, and chitin, while hyperthermophilic marine bacteria (T(opt)>or=80 degrees C) utilize the same glucans as well as hemicellulose, such as xylans and mannans. However, none of these organisms are able to efficiently utilize crystalline cellulose. Among the thermophiles, this ability is limited to a few terrestrial bacteria with upper temperature limits for growth near 75 degrees C. Deconstruction of crystalline cellulose by these extreme thermophiles is achieved by 'free' primary cellulases, which are distinct from those typically associated with large multi-enzyme complexes known as cellulosomes. These primary cellulases also differ from the endoglucanases (referred to here as 'secondary cellulases') reported from marine hyperthermophiles that show only weak activity toward cellulose. Many extremely thermophilic enzymes implicated in the deconstruction of lignocellulose can be identified in genome sequences, and many more promising biocatalysts probably remain annotated as 'hypothetical proteins'. Characterization of these enzymes will require intensive effort but is likely to generate new opportunities for the use of renewable resources as biofuels.

  5. Thermal and Spectroscopic Characterization of a Proton Pumping Rhodopsin from an Extreme Thermophile*

    PubMed Central

    Tsukamoto, Takashi; Inoue, Keiichi; Kandori, Hideki; Sudo, Yuki

    2013-01-01

    So far retinylidene proteins (∼rhodopsin) have not been discovered in thermophilic organisms. In this study we investigated and characterized a microbial rhodopsin derived from the extreme thermophilic bacterium Thermus thermophilus, which lives in a hot spring at around 75 °C. The gene for the retinylidene protein, named thermophilic rhodopsin (TR), was chemically synthesized with codon optimization. The codon optimized TR protein was functionally expressed in the cell membranes of Escherichia coli cells and showed active proton transport upon photoillumination. Spectroscopic measurements revealed that the purified TR bound only all-trans-retinal as a chromophore and showed an absorption maximum at 530 nm. In addition, TR exhibited both photocycle kinetics and pH-dependent absorption changes, which are characteristic of rhodopsins. Of note, time-dependent thermal denaturation experiments revealed that TR maintained its absorption even at 75 °C, and the denaturation rate constant of TR was much lower than those of other proton pumping rhodopsins such as archaerhodopsin-3 (200 ×), Haloquadratum walsbyi bacteriorhodopsin (by 10-times), and Gloeobacter rhodopsin (100 ×). Thus, these results suggest that microbial rhodopsins are also distributed among thermophilic organisms and have high stability. TR should allow the investigation of the molecular mechanisms of ion transport and protein folding. PMID:23740255

  6. Thermal and spectroscopic characterization of a proton pumping rhodopsin from an extreme thermophile.

    PubMed

    Tsukamoto, Takashi; Inoue, Keiichi; Kandori, Hideki; Sudo, Yuki

    2013-07-26

    So far retinylidene proteins (∼rhodopsin) have not been discovered in thermophilic organisms. In this study we investigated and characterized a microbial rhodopsin derived from the extreme thermophilic bacterium Thermus thermophilus, which lives in a hot spring at around 75 °C. The gene for the retinylidene protein, named thermophilic rhodopsin (TR), was chemically synthesized with codon optimization. The codon optimized TR protein was functionally expressed in the cell membranes of Escherichia coli cells and showed active proton transport upon photoillumination. Spectroscopic measurements revealed that the purified TR bound only all-trans-retinal as a chromophore and showed an absorption maximum at 530 nm. In addition, TR exhibited both photocycle kinetics and pH-dependent absorption changes, which are characteristic of rhodopsins. Of note, time-dependent thermal denaturation experiments revealed that TR maintained its absorption even at 75 °C, and the denaturation rate constant of TR was much lower than those of other proton pumping rhodopsins such as archaerhodopsin-3 (200 ×), Haloquadratum walsbyi bacteriorhodopsin (by 10-times), and Gloeobacter rhodopsin (100 ×). Thus, these results suggest that microbial rhodopsins are also distributed among thermophilic organisms and have high stability. TR should allow the investigation of the molecular mechanisms of ion transport and protein folding.

  7. State of the art review of biofuels production from lignocellulose by thermophilic bacteria.

    PubMed

    Jiang, Yujia; Xin, Fengxue; Lu, Jiasheng; Dong, Weiliang; Zhang, Wenming; Zhang, Min; Wu, Hao; Ma, Jiangfeng; Jiang, Min

    2017-12-01

    Biofuels, including ethanol and butanol, are mainly produced by mesophilic solventogenic yeasts and Clostridium species. However, these microorganisms cannot directly utilize lignocellulosic materials, which are abundant, renewable and non-compete with human demand. More recently, thermophilic bacteria show great potential for biofuels production, which could efficiently degrade lignocellulose through the cost effective consolidated bioprocessing. Especially, it could avoid contamination in the whole process owing to its relatively high fermentation temperature. However, wild types thermophiles generally produce low levels of biofuels, hindering their large scale production. This review comprehensively summarizes the state of the art development of biofuels production by reported thermophilic microorganisms, and also concludes strategies to improve biofuels production including the metabolic pathways construction, co-culturing systems and biofuels tolerance. In addition, strategies to further improve butanol production are proposed. Copyright © 2017 Elsevier Ltd. All rights reserved.

  8. Characterization of Hydrolysis Kinetics in Staged Anaerobic Digestion of Wastewater Treatment Sludge.

    PubMed

    Zamanzadeh, Mirzaman; Parker, Wayne J

    2018-01-01

      The hydrolysis of mixed primary and secondary sludges in two-stage anaerobic digestion was evaluated and compared with conventional single-stage digestion, using various temperature-phased configurations of M1-M2, M1-T3, T1-T2, and T1-M3. A dual hydrolysis model best described the hydrolysis in all tests. This model was also able to consistently estimate the readily and slowly fractions of particulate chemical oxygen demand (COD) of raw sludge used in the tests. The hydrolysis kinetic coefficients (Khyd_s and Khyd_r) estimated for the mesophilic digesters were significantly greater in the short hydraulic retention time (HRT) M1 digester than those of the extended HRT digesters. Conversely, at thermophilic temperatures only Khyd_r was greater in short HRT T1 digester when compared to the extended HRT digesters. The increased Khyd_r and reduced Khyd_s values due to staging effect were explained with surface reaction models and endogenous decay. The temperature dependency of Khyd_s and Khyd_r was also explored in the staged digesters.

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

    PubMed

    Jensen, T R; Lastra Milone, T; Petersen, G; Andersen, H R

    2017-04-01

    Anaerobic hydrolysis in activated return sludge was investigated in laboratory scale experiments to find if intermittent aeration would accelerate anaerobic hydrolysis rates compared to anaerobic hydrolysis rates under strict anaerobic conditions. The intermittent reactors were set up in a 240 h experiment with intermittent aeration (3 h:3 h) in a period of 24 h followed by a subsequent anaerobic period of 24 h in a cycle of 48 h which was repeated five times during the experiment. The anaerobic reactors were kept under strict anaerobic conditions in the same period (240 h). Two methods for calculating hydrolysis rates based on soluble chemical oxygen demand were compared. Two-way analysis of variance with the Bonferroni post-test was performed in order to register any significant difference between reactors with intermittent aeration and strictly anaerobic conditions respectively. The experiment demonstrated a statistically significant difference in favor of the reactors with intermittent aeration showing a tendency towards accelerated anaerobic hydrolysis rates due to application of intermittent aeration. The conclusion of the work is thus that intermittent aeration applied in the activated return sludge process can improve the treatment capacity further in full scale applications.

  10. Mechanisms Used for Genomic Proliferation by Thermophilic Group II Introns

    PubMed Central

    Mohr, Georg; Ghanem, Eman; Lambowitz, Alan M.

    2010-01-01

    Mobile group II introns, which are found in bacterial and organellar genomes, are site-specific retroelments hypothesized to be evolutionary ancestors of spliceosomal introns and retrotransposons in higher organisms. Most bacteria, however, contain no more than one or a few group II introns, making it unclear how introns could have proliferated to higher copy numbers in eukaryotic genomes. An exception is the thermophilic cyanobacterium Thermosynechococcus elongatus, which contains 28 closely related copies of a group II intron, constituting ∼1.3% of the genome. Here, by using a combination of bioinformatics and mobility assays at different temperatures, we identified mechanisms that contribute to the proliferation of T. elongatus group II introns. These mechanisms include divergence of DNA target specificity to avoid target site saturation; adaptation of some intron-encoded reverse transcriptases to splice and mobilize multiple degenerate introns that do not encode reverse transcriptases, leading to a common splicing apparatus; and preferential insertion within other mobile introns or insertion elements, which provide new unoccupied sites in expanding non-essential DNA regions. Additionally, unlike mesophilic group II introns, the thermophilic T. elongatus introns rely on elevated temperatures to help promote DNA strand separation, enabling access to a larger number of DNA target sites by base pairing of the intron RNA, with minimal constraint from the reverse transcriptase. Our results provide insight into group II intron proliferation mechanisms and show that higher temperatures, which are thought to have prevailed on Earth during the emergence of eukaryotes, favor intron proliferation by increasing the accessibility of DNA target sites. We also identify actively mobile thermophilic introns, which may be useful for structural studies, gene targeting in thermophiles, and as a source of thermostable reverse transcriptases. PMID:20543989

  11. Environmental assessment of anaerobically digested sludge reuse in agriculture: potential impacts of emerging micropollutants.

    PubMed

    Hospido, Almudena; Carballa, Marta; Moreira, Maite; Omil, Francisco; Lema, Juan M; Feijoo, Gumersindo

    2010-05-01

    Agricultural application of sewage sludge has been emotionally discussed in the last decades, because the latter contains organic micropollutants with unknown fate and risk potential. In this work, the reuse of anaerobically digested sludge in agriculture is evaluated from an environmental point of view by using Life Cycle Assessment methodology. More specifically, the potential impacts of emerging micropollutants, such as pharmaceuticals and personal care products, present in the sludge have been quantified. Four scenarios were considered according to the temperature of the anaerobic digestion (mesophilic or thermophilic) and the sludge retention time (20 or 10d), and they have been compared with the non-treated sludge. From an environmental point of view, the disposal of undigested sludge is not the most suitable alternative, except for global warming due to the dominance (65-85%) of the indirect emissions associated to the electricity use. Nutrient-related direct emissions dominate the eutrophication category impact in all the scenarios (>71.4%), although a beneficial impact related to the avoidance of industrial fertilisers production is also quantified (up to 6.7%). In terms of human and terrestrial toxicity, the direct emissions of heavy metals to soil dominate these two impact categories (>70%), and the contribution of other micropollutants is minimal. Moreover, only six (Galaxolide, Tonalide, Diazepam, Ibuprofen, Sulfamethoxazole and 17alpha-ethinyloestradiol) out of the 13 substances considered are really significant since they account for more than 95% of the overall micropollutants impact.

  12. [Agroindustrial wastes methanization and bacterial composition in anaerobic digestion].

    PubMed

    González-Sánchez, María E; Pérez-Fabiel, Sergio; Wong-Villarreal, Arnoldo; Bello-Mendoza, Ricardo; Yañez-Ocampo, Gustavo

    2015-01-01

    The tons of organic waste that are annually generated by agro-industry, can be used as raw material for methane production. For this reason, it is important to previously perform biodegradability tests to organic wastes for their full scale methanization. This paper addresses biodegradability, methane production and the behavior of populations of eubacteria and archaeabacteria during anaerobic digestion of banana, mango and papaya agroindustrial wastes. Mango and banana wastes had higher organic matter content than papaya in terms of their volatile solids and total solid rate (94 and 75% respectively). After 63 days of treatment, the highest methane production was observed in banana waste anaerobic digestion: 63.89ml CH4/per gram of chemical oxygen demand of the waste. In the PCR-DGGE molecular analysis, different genomic footprints with oligonucleotides for eubacteria and archeobacteria were found. Biochemical methane potential results proved that banana wastes have the best potential to be used as raw material for methane production. The result of a PCR- DGGE analysis using specific oligonucleotides enabled to identify the behavior of populations of eubacteria and archaeabacteria present during the anaerobic digestion of agroindustrial wastes throughout the process. Copyright © 2015 Asociación Argentina de Microbiología. Publicado por Elsevier España, S.L.U. All rights reserved.

  13. Anaerobic bacteria

    MedlinePlus

    Anaerobic bacteria are bacteria that do not live or grow when oxygen is present. In humans, these bacteria ... Brook I. Diseases caused by non-spore-forming anaerobic bacteria. In: Goldman L, Schafer AI, eds. Goldman-Cecil ...

  14. Significance of dissolved methane in effluents of anaerobically ...

    EPA Pesticide Factsheets

    The need for energy efficient Domestic Wastewater (DWW) treatment is increasing annually with population growth and expanding global energy demand. Anaerobic treatment of low strength DWW produces methane which can be used to as an energy product. Temperature sensitivity, low removal efficiencies (Chemical Oxygen Demand (COD), Suspended Solids (SS), and Nutrients), alkalinity demand, and potential greenhouse gas (GHG) emissions have limited its application to warmer climates. Although well designed anaerobic Membrane Bioreactors (AnMBRs) are able to effectively treat DWW at psychrophilic temperatures (10–30 °C), lower temperatures increase methane solubility leading to increased energy losses in the form of dissolved methane in the effluent. Estimates of dissolved methane losses are typically based on concentrations calculated using Henry's Law but advection limitations can lead to supersaturation of methane between 1.34 and 6.9 times equilibrium concentrations and 11–100% of generated methane being lost in the effluent. In well mixed systems such as AnMBRs which use biogas sparging to control membrane fouling, actual concentrations approach equilibrium values. Non-porous membranes have been used to recover up to 92.6% of dissolved methane and well suited for degassing effluents of Upflow Anaerobic Sludge Blanket (UASB) reactors which have considerable solids and organic contents and can cause pore wetting and clogging in microporous membrane modules. Micro

  15. Effect of thermal additions on the density and distribution of thermophilic amoebae and pathogenic Naegleria fowleri in a newly created cooling lake

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Tyndall, R.L.; Ironside, K.S.; Metler, P.L.

    1989-03-01

    Pathogenic Naegleria fowleri is the causative agent of fatal human amoebic meningoencephalitis. The protozoan is ubiquitous in nature, and its presence is enhanced by thermal additions. In this investigation, water and sediments from a newly created cooling lake were quantitatively analyzed for the presence of thermophilic amoebae, thermophilic Naegleria spp., and the pathogen Naegleria fowleri. During periods of thermal additions, the concentrations of thermophilic amoebae and thermophilic Naegleria spp. increased as much as 5 orders of magnitude, and the concentration of the pathogen N. fowleri increased as much as 2 orders of magnitude. Concentrations of amoebae returned to prior thermalmore » perturbation levels within 30 to 60 days after cessation of thermal additions. Increases in the thermophilic amoeba concentrations were noted in Savannah River oxbows downriver from the Savannah River plant discharge streams as compared with oxbows upriver from the discharges. Concentrations of thermophilic amoebae and thermophilic Naegleria spp. correlated significantly with temperature and conductivity. Air samples taken proximal to the lade during periods of thermal addition showed no evidence of thermophilic Naegleria spp. Isoenzyme patterns of the N. fowleri isolated from the cooling lake were identical to patterns of N. fowleri isolated from other sites in the United States and Belgium.« less

  16. Leaching: use of a thermophilic and chemoautotrophic microbe.

    PubMed

    Brierley, C L; Murr, L E

    1973-02-02

    A chemoautotrophic, thermophilic, and acidophilic microorganism capable of oxidizing reduced sulfur and iron compounds and leaching concentrates of molybdenite and chalcopyrite at 60 degrees C has been characterized by transmission and scanning electron microscopy. This constitutes the first direct observations of microorganisms on ore fines.

  17. [Isolation and characterization of Thermopirellula anaerolimosa gen. nov., sp. nov., an obligate anaerobic hydrogen-producing bacterium of the phylum Planctomycetes].

    PubMed

    Liu, Dongying; Liu, Yi; Men, Xuehui; Guo, Qunqun; Guo, Rongbo; Qiu, Yanling

    2012-08-04

    To cultivate various yet-to-be cultured heterotrophs from anaerobic granule sludge, we used a selective culture medium with low concentrations of substrates supplemented a variety of antibiotics. An obligate anaerobic, thermophilic, hydrogen-producing bacterium, strain VM20-7(T), was isolated from an upflow anaerobic sludge blanket (UASB) reactor treating high-strength organic wastewater from isomerized sugar production processes. Cells of strain VM20-7(T) are non-motile, spherical, pear or teardrop shaped, occurring singly(o)r as aggregates (0.7 - 2.0 microm x 0.7 - 2.0 microm). Spore formation was not observed. Growth temperature ranges from 35 - 50 degrees C (optimum 45 degrees C), pH ranges from 6.0 - 8.3 (optimum 7.0 - 7.5) , NaCl tolerant concentration ranges from 0% - 0.5% (w/v, optimum 0% ). Nitrate, sulfate, thiosulfate, sulfite, elemental sulfur and Fe (III)-NTA were not used as terminal electron acceptors. Strain VM20-7(T) utilizes a wide range of carbohydrates, including glucose, maltose, ribose, xylose, sucrose, galactose, mannose, raffinose, pectin, yeast extract and xylan. Acetate and H2 are the main end products of glucose fermentation. The G + C content of the genomic DNA was 60.9 mol%. 16S rRNA gene sequence analysis revealed that it is related to the Pirellula-Rhodopirellula-Blastopirellula (PRB) clade within the order Planctomycetales (82.7 - 84.3% similarity with 16S rRNA genes of other known related species). The first obligate anaerobic bacterium within the phylum Planctomycetes was isolated with low concentration of carbohydrates and antibiotics. On the basis of the physiological and phylogenetic data, the name Thermopirellula anaerolimosa gen. nov. , sp. nov. is proposed for strain VM20-7(T) (= CGMCC 1.5169(T) = JCM 17478(T) = DSM 24165(T)).

  18. Fate of antibiotic and metal resistance genes during two-phase anaerobic digestion of residue sludge revealed by metagenomic approach.

    PubMed

    Wu, Ying; Cui, Erping; Zuo, Yiru; Cheng, Weixiao; Chen, Hong

    2018-05-01

    The prevalence and persistence of antibiotic resistance genes in wastewater treatment plants (WWTPs) is of growing interest, and residual sludge is among the main sources for the release of antibiotic resistance genes (ARGs). Moreover, heavy metals concentrated in dense microbial communities of sludge could potentially favor co-selection of ARGs and metal resistance genes (MRGs). Residual sludge treatment is needed to limit the spread of resistance from WWTPs into the environment. This study aimed to explore the fate of ARGs and MRGs during thermophilic two-phase (acidogenic/methanogenic phase) anaerobic digestion by metagenomic analysis. The occurrence and abundance of mobile genetic elements were also determined based on the SEED database. Among the 27 major ARG subtypes detected in feed sludge, large reductions (> 50%) in 6 ARG subtypes were achieved by acidogenic phase (AP), while 63.0% of the ARG subtypes proliferated in the following methanogenic phase (MP). In contrast, a 2.8-fold increase in total MRG abundance was found in AP, while the total abundance during MP decreased to the same order of magnitude as in feed sludge. The distinct dynamics of ARGs and MRGs during the two-phase anaerobic digestion are noteworthy, and more specific treatments are required to limit their proliferation in the environment.

  19. Thermal adaptation analyzed by comparison of protein sequences from mesophilic and extremely thermophilic Methanococcus species

    NASA Technical Reports Server (NTRS)

    Haney, P. J.; Badger, J. H.; Buldak, G. L.; Reich, C. I.; Woese, C. R.; Olsen, G. J.

    1999-01-01

    The genome sequence of the extremely thermophilic archaeon Methanococcus jannaschii provides a wealth of data on proteins from a thermophile. In this paper, sequences of 115 proteins from M. jannaschii are compared with their homologs from mesophilic Methanococcus species. Although the growth temperatures of the mesophiles are about 50 degrees C below that of M. jannaschii, their genomic G+C contents are nearly identical. The properties most correlated with the proteins of the thermophile include higher residue volume, higher residue hydrophobicity, more charged amino acids (especially Glu, Arg, and Lys), and fewer uncharged polar residues (Ser, Thr, Asn, and Gln). These are recurring themes, with all trends applying to 83-92% of the proteins for which complete sequences were available. Nearly all of the amino acid replacements most significantly correlated with the temperature change are the same relatively conservative changes observed in all proteins, but in the case of the mesophile/thermophile comparison there is a directional bias. We identify 26 specific pairs of amino acids with a statistically significant (P < 0.01) preferred direction of replacement.

  20. Evolution of butyric acid and the methanogenic microbial population in a thermophilic dry anaerobic reactor.

    PubMed

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

    2010-10-01

    Knowledge of the relationship between the evolution of butyric acid, the main precursor of methane, and the methanogenic microbial population, quantified by fluorescent in situ hybridization (FISH), is important for understating and controlling the anaerobic digestion of solid waste. In this study, a statistical analysis has been made of this relationship in a laboratory-scale continuously stirred tank reactor (CSTR) without recycling of biomass. The CSTR has been operated at four organic loading rates (OLR) between 4.42 and 7.50 kg VS m(-3)day(-1), using the Organic Fraction of Municipal Solid Waste (OFMSW) as the substrate. During the start-up stage, the consumption of butyric acid is related to the population of H(2)-utilising methanogens (R(2)=0.774), whereas in the stabilization stage, it is related to that of acetoclastic methanogens (R(2)=0.716). Therefore, it can be concluded that the methanogenic population dynamics are closely correlated with the concentration of volatile fatty acids, and in particular, with that of butyric acid. (c) 2010 Elsevier Ltd. All rights reserved.

  1. Prevalence and strain diversity of thermophilic campylobacters in cattle, sheep and swine farms.

    PubMed

    Oporto, B; Esteban, J I; Aduriz, G; Juste, R A; Hurtado, A

    2007-10-01

    To determine prevalence and strain diversity of thermophilic campylobacters in healthy ruminants and swine. Faecal samples collected from 343 herds (120 sheep, 124 beef cattle, 82 dairy cattle and 17 swine) in the Basque Country were screened in pools for thermophilic campylobacters. Two hundred and three herds were positive (67.1% dairy cattle, 58.9% beef cattle, 55.0% sheep and 52.9% pig), and species-specific PCR identified Campylobacter jejuni in 20.7% of the herds and Campylobacter coli in 6.4%. Campylobacter coli was isolated from the four production systems and was the most prevalent species in swine, where C. jejuni was not found. Other thermophilic campylobacters were found in all production systems. Four hundred and ninety-three animals from 11 positive herds were individually analysed, detecting significantly higher within-herd prevalences in dairy cattle (66.7%) and swine (57.8%) than in sheep (8.8%) or beef cattle (5.4%). flaA PCR-RFLP and pulsed-field gel electrophoresis analysis of a selection of isolates showed high genetic diversity. Healthy swine, cattle and sheep are important reservoirs of thermophilic campylobacters of different species and high genetic diversity. Efficient farm-based intervention measures are needed to reduce risk of infection. Non-C. jejuni/C. coli species should be monitored to investigate their significance for infection.

  2. Differences in the catalytic mechanisms of mesophilic and thermophilic indole-3-glycerol phosphate synthase enzymes at their adaptive temperatures.

    PubMed

    Zaccardi, Margot J; Mannweiler, Olga; Boehr, David D

    2012-02-10

    Thermophilic enzymes tend to be less catalytically-active at lower temperatures relative to their mesophilic counterparts, despite having very similar crystal structures. An often cited hypothesis for this general observation is that thermostable enzymes have evolved a more rigid tertiary structure in order to cope with their more extreme, natural environment, but they are also less flexible at lower temperatures, leading to their lower catalytic activity under mesophilic conditions. An alternative hypothesis, however, is that complementary thermophilic-mesophilic enzyme pairs simply operate through different evolutionary-optimized catalytic mechanisms. In this communication, we present evidence that while the steps of the catalytic mechanisms for mesophilic and thermophilic indole-3-glycerol phosphate synthase (IGPS) enzymes are fundamentally similar, the identity of the rate-determining step changes as a function of temperature. Our findings indicate that while product release is rate-determining at 25°C for thermophilic IGPS, near its adaptive temperature (75°C), a proton transfer event, involving a general acid, becomes rate-determining. The rate-determining steps for thermophilic and mesophilic IGPS enzymes are also different at their respective, adaptive temperatures with the mesophilic IGPS-catalyzed reaction being rate-limited before irreversible CO2 release, and the thermophilic IGPS-catalyzed reaction being rate limited afterwards. Copyright © 2012 Elsevier Inc. All rights reserved.

  3. Economic and environmental analysis of four different configurations of anaerobic digestion for food waste to energy conversion using LCA for: a food service provider case study.

    PubMed

    Franchetti, Matthew

    2013-07-15

    The US disposes of more than 34 million tons of food waste in landfills per year. As this food waste decomposes it generates methane gas and negatively contributes to global warming. Diverting theses organic food wastes from landfills and to emerging technologies will prevent these wastes and greenhouse gas emissions while at the same time generating a source renewable energy by collecting the emitted gases. From a waste prevention standpoint, instead of the food waste decomposing at local landfills, it is being converted into an energy source and the by-product may be used as a fertilizer (Fine and Hadas, 2012). The purpose of this study was to compare four different configurations of anaerobic digestion of organic waste to energy technologies from an economic, energy, and emissions standpoint using LCA via a case study at a large food services provider in Northwest Ohio, USA. The technologies studied included two-stage anaerobic digestion system using ultrasound pre-treating, two stage continuous combined thermophilic acidogenic hydrogenesis and mesophilic with recirculation of the digested sludge, long-term anaerobic digestion of food waste stabilized by trace elements, and single stage anaerobic digestion. Using LCA, these scenarios were compared to landfill disposal of the food waste. The findings from the case study indicated that implementing on-site waste to energy systems will result in lower operation costs and lower environmental impacts. In addition, a standardized environmental and economic comparison of competing food waste to energy technologies is provided. Copyright © 2013 Elsevier Ltd. All rights reserved.

  4. Response of anaerobes to methyl fluoride, 2-bromoethanesulfonate and hydrogen during acetate degradation.

    PubMed

    Hao, Liping; Lü, Fan; Li, Lei; Shao, Liming; He, Pinjing

    2013-05-01

    To use the selective inhibition method for quantitative analysis of acetate metabolism in methanogenic systems, the responses of microbial communities and metabolic activities, which were involved in anaerobic degradation of acetate, to the addition of methyl fluoride (CH3F), 2-bromoethanesulfonate (BES) and hydrogen were investigated in a thermophilic batch experiment. Both the methanogenic inhibitors, i.e., CH3F and BES, showed their effectiveness on inhibiting CH4 production, whereas acetate metabolism other than acetoclastic methanogenesis was stimulated by BES, as reflected by the fluctuated acetate concentration. Syntrophic acetate oxidation was thermodynamically blocked by hydrogen (H2), while H2-utilizing reactions as hydrogenotrophic methanogenesis and homoacetogenesis were correspondingly promoted. Results of PCR-DGGE fingerprinting showed that, CH3F did not influence the microbial populations significantly. However, the BES and hydrogen notably altered the bacterial community structures and increased the diversity. BES gradually changed the methanogenic community structure by affecting the existence of different populations to different levels, whilst H2 greatly changed the abundance of different methanogenic populations, and induced growth of new species.

  5. Chemical and microbial changes during autothermal thermophilic aerobic digestion (ATAD) of sewage sludge.

    PubMed

    Liu, Shugen; Song, Fanyong; Zhu, Nanwen; Yuan, Haiping; Cheng, Jiehong

    2010-12-01

    Autothermal thermophilic aerobic digestion (ATAD) is a promising process for sewage sludge stabilization. Batch experiments were conducted on sewage sludge collected from a municipal wastewater treatment plant in Shanghai, China, to evaluate the effectiveness of the ATAD system by determining changes in volatile suspended solids (VSSs) and to study its microbial diversity by denaturing gradient gel electrophoresis of 16S rRNA gene sequences amplified by PCR. The digestion system achieved rapid degradation of the organic substrate at 55 degrees C. The VSS was removed by up to 45.3% and 50.4% at 216 h and 264 h, respectively, while NH(4)(+)-N, chemical oxidation demand and total organic carbon of supernatant as well as total nitrogen did not exhibit obvious declines after 168 h. The microbial diversity changed during the thermophilic process as thermophiles belonging to the Hydrogenophilaceae, Thermotogaceae, Clostridiaceae and the genus Ureibacillus replaced less temperature-tolerant microorganisms such as Sphingobacteriaceae and the genus Trichococcus. Copyright (c) 2010 Elsevier Ltd. All rights reserved.

  6. Improved fermentation performance in an expanded ectopic fermentation system inoculated with thermophilic bacteria.

    PubMed

    Guo, Hui; Zhu, Changxiong; Geng, Bing; Liu, Xue; Ye, Jing; Tian, Yunlong; Peng, Xiawei

    2015-12-01

    Previous research showed that ectopic fermentation system (EFS) inoculated with thermophilic bacteria is an excellent alternative for cow wastewater treatment. In this study, the effects of thermophilic bacterial consortium on the efficiency and quality of the fermentation process in EFS were evaluated by measuring physicochemical and environmental factors and the changes in organic matter composition. In parallel, the microbial communities correlated with fermentation performance were identified. Inoculation of EFS with thermophilic bacterial consortium led to higher temperatures, increased wastewater requirements for continuous fermentation, and improved quality of the litters in terms of physicochemical factors, security test, functional group analysis, and bacterial community composition. The relationship between the transformation of organic component and the dominant bacteria species indicated that environmental factors contributed to strain growth, which subsequently promoted the fermentation process. The results highlight the great potential of EFS model for wide application in cow wastewater treatment and re-utilization as bio-fertilizer. Copyright © 2015 Elsevier Ltd. All rights reserved.

  7. Anaerobic Digestion and its Applications

    EPA Science Inventory

    Anaerobic digestion is a natural biological process. The initials "AD" may refer to the process of anaerobic digestion, or the built systems of anaerobic digesters. While there are many kinds of digesters, the biology is basically the same for all. Anaerobic digesters are built...

  8. Anaerobic Biodegradation of Detergent Surfactants

    PubMed Central

    Merrettig-Bruns, Ute; Jelen, Erich

    2009-01-01

    Detergent surfactants can be found in wastewater in relevant concentrations. Most of them are known as ready degradable under aerobic conditions, as required by European legislation. Far fewer surfactants have been tested so far for biodegradability under anaerobic conditions. The natural environment is predominantly aerobic, but there are some environmental compartments such as river sediments, sub-surface soil layer and anaerobic sludge digesters of wastewater treatment plants which have strictly anaerobic conditions. This review gives an overview on anaerobic biodegradation processes, the methods for testing anaerobic biodegradability, and the anaerobic biodegradability of different detergent surfactant types (anionic, nonionic, cationic, amphoteric surfactants).

  9. Isolation and Screening of Thermophilic Bacilli from Compost for Electrotransformation and Fermentation: Characterization of Bacillus smithii ET 138 as a New Biocatalyst

    PubMed Central

    Bosma, Elleke F.; van de Weijer, Antonius H. P.; Daas, Martinus J. A.; van der Oost, John; de Vos, Willem M.

    2015-01-01

    Thermophilic bacteria are regarded as attractive production organisms for cost-efficient conversion of renewable resources to green chemicals, but their genetic accessibility is a major bottleneck in developing them into versatile platform organisms. In this study, we aimed to isolate thermophilic, facultatively anaerobic bacilli that are genetically accessible and have potential as platform organisms. From compost, we isolated 267 strains that produced acids from C5 and C6 sugars at temperatures of 55°C or 65°C. Subsequently, 44 strains that showed the highest production of acids were screened for genetic accessibility by electroporation. Two Geobacillus thermodenitrificans isolates and one Bacillus smithii isolate were found to be transformable with plasmid pNW33n. Of these, B. smithii ET 138 was the best-performing strain in laboratory-scale fermentations and was capable of producing organic acids from glucose as well as from xylose. It is an acidotolerant strain able to produce organic acids until a lower limit of approximately pH 4.5. As genetic accessibility of B. smithii had not been described previously, six other B. smithii strains from the DSMZ culture collection were tested for electroporation efficiencies, and we found the type strain DSM 4216T and strain DSM 460 to be transformable. The transformation protocol for B. smithii isolate ET 138 was optimized to obtain approximately 5 × 103 colonies per μg plasmid pNW33n. Genetic accessibility combined with robust acid production capacities on C5 and C6 sugars at a relatively broad pH range make B. smithii ET 138 an attractive biocatalyst for the production of lactic acid and potentially other green chemicals. PMID:25556192

  10. In-situ biogas upgrading during anaerobic digestion of food waste amended with walnut shell biochar at bench scale.

    PubMed

    Linville, Jessica L; Shen, Yanwen; Ignacio-de Leon, Patricia A; Schoene, Robin P; Urgun-Demirtas, Meltem

    2017-06-01

    A modified version of an in-situ CO 2 removal process was applied during anaerobic digestion of food waste with two types of walnut shell biochar at bench scale under batch operating mode. Compared with the coarse walnut shell biochar, the fine walnut shell biochar has a higher ash content (43 vs. 36 wt%) and higher concentrations of calcium (31 vs. 19 wt% of ash), magnesium (8.4 vs. 5.6 wt% of ash) and sodium (23.4 vs. 0.3 wt% of ash), but a lower potassium concentration (0.2 vs. 40% wt% of ash). The 0.96-3.83 g biochar (g VS added ) -1 fine walnut shell biochar amended digesters produced biogas with 77.5%-98.1% CH 4 content by removing 40%-96% of the CO 2 compared with the control digesters at mesophilic and thermophilic temperature conditions. In a direct comparison at 1.83 g biochar (g VS added ) -1 , the fine walnut shell biochar amended digesters (85.7% CH 4 content and 61% CO 2 removal) outperformed the coarse walnut shell biochar amended digesters (78.9% CH 4 content and 51% CO 2 removal). Biochar addition also increased alkalinity as CaCO 3 from 2800 mg L -1 in the control digesters to 4800-6800 mg L -1 , providing process stability for food waste anaerobic digestion.

  11. Chlorophyll a with a farnesyl tail in thermophilic cyanobacteria.

    PubMed

    Wiwczar, Jessica M; LaFountain, Amy M; Wang, Jimin; Frank, Harry A; Brudvig, Gary W

    2017-11-01

    Photosystem II (PSII) of oxygenic photosynthetic organisms normally contains exclusively chlorophyll a (Chl a) as its major light-harvesting pigment. Chl a canonically consists of the chlorin headgroup with a 20-carbon, 4-isoprene unit, phytyl tail. We have examined the 1.9 Å crystal structure of PSII from thermophilic cyanobacteria reported by Shen and coworkers in 2012 (PDB accession of 3ARC/3WU2). A newly refined electron density map from this structure, presented here, reveals that some assignments of the cofactors may be different from those modeled in the 3ARC/3WU2 structure, including a specific Chl a that appears to have a truncated tail by one isoprene unit. We provide experimental evidence using high-performance liquid chromatography and mass spectrometry for a small population of Chl a esterified to a 15-carbon farnesyl tail in PSII of thermophilic cyanobacteria.

  12. [Current clinical significance of anaerobic bacteremia].

    PubMed

    Jirsa, Roman; Marešová, Veronika; Brož, Zdeněk

    2010-10-01

    to estimate tje current clinical significance of anaerobic bacteremia in a group of Czech hospitals. this retrospective analysis comprised 8 444 anaerobic blood cultures in patients admitted to four Czech hospitals between 2004 and 2007. in 16 patients, blood cultures yielded significant anaerobic bacteria. Thus, anaerobic bacteremia accounted for less than 2 % of clinically significant bacteremia. Four patients (18 %) died but none of the deaths could be clearly attributable to anaerobic bacteria in the bloodstream. The most common comorbidities predisposing to anaerobic bacteremia and the most frequent sources of infection were similar to those reported by other authors. The majority of anaerobic bacteremia cases were due to gram-negative bacteria, followed by Clostridium perfringens and, surprisingly, Eubacterium spp. (particularly Eubacterium lentum). anaerobic bacteremia remains rare. The comparison of our data with those by other authors suggests that (despite the reported high mortality) the actual clinical significance of anaerobic bacteremia is rather controversial and that the anaerobic bacteremia might not correspond to more serious pathogenic role of the anaerobic bacteria as the source of infection.

  13. Transformable facultative thermophile Geobacillus stearothermophilus NUB3621 as a host strain for metabolic engineering

    PubMed Central

    Blanchard, Kristen; Robic, Srebrenka

    2014-01-01

    Metabolic engineers develop inexpensive enantioselective syntheses of high-value compounds, but their designs are sometimes confounded by the misfolding of heterologously expressed proteins. Geobacillus stearothermophilus NUB3621 is a readily transformable facultative thermophile. It could be used to express and properly fold proteins derived from its many mesophilic or thermophilic Bacillaceae relatives or to direct the evolution of thermophilic variants of mesophilic proteins. Moreover, its capacity for high-temperature growth should accelerate chemical transformation rates in accordance with the Arrhenius equation and reduce the risks of microbial contamination. Its tendency to sporulate in response to nutrient depletion lowers the costs of storage and transportation. Here, we present a draft genome sequence of G. stearothermophilus NUB3621 and describe inducible and constitutive expression plasmids that function in this organism. These tools will help us and others to exploit the natural advantages of this system for metabolic engineering applications. PMID:24788326

  14. Evaluation of dairy powder products implicates thermophilic sporeformers as the primary organisms of interest.

    PubMed

    Watterson, M J; Kent, D J; Boor, K J; Wiedmann, M; Martin, N H

    2014-01-01

    Dairy powder products (e.g., sweet whey, nonfat dry milk, acid whey, and whey protein concentrate-80) are of economic interest to the dairy industry. According to the US Dairy Export Council, customers have set strict tolerances (<500 to <1,000/g) for thermophilic and mesophilic spores in dairy powders; therefore, understanding proliferation and survival of sporeforming organisms within dairy powder processing plants is necessary to control and reduce sporeformer counts. Raw, work-in-process, and finished product samples were collected from 4 dairy powder processing facilities in the northeastern United States over a 1-yr period. Two separate spore treatments: (1) 80°C for 12min (to detect sporeformers) and (2) 100°C for 30min (to detect highly heat resistant sporeformers) were applied to samples before microbiological analyses. Raw material, work-in-process, and finished product samples were analyzed for thermophilic, mesophilic, and psychrotolerant sporeformers, with 77.5, 71.0, and 4.6% of samples being positive for those organisms, respectively. Work-in-process and finished product samples were also analyzed for highly heat resistant thermophilic and mesophilic sporeformers, with 63.7 and 42.6% of samples being positive, respectively. Sporeformer prevalence and counts varied considerably by product and plant; sweet whey and nonfat dry milk showed a higher prevalence of thermophilic and mesophilic sporeformers compared with acid whey and whey protein concentrate-80. Unlike previous reports, we found limited evidence for increased spore counts toward the end of processing runs. Our data provide important insight into spore contamination patterns associated with production of different types of dairy powders and support that thermophilic sporeformers are the primary organism of concern in dairy powders. Copyright © 2014 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

  15. Proteogenomic Analysis of a Thermophilic Bacterial Consortium Adapted to Deconstruct Switchgrass

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    D'haeseleer, Patrik; Gladden, John M.; Allgaier, Martin

    2013-07-19

    Thermophilic bacteria are a potential source of enzymes for the deconstruction of lignocellulosic biomass. However, the complement of proteins used to deconstruct biomass and the specific roles of different microbial groups in thermophilic biomass deconstruction are not well-explored. Here we report on the metagenomic and proteogenomic analyses of a compost-derived bacterial consortium adapted to switchgrass at elevated temperature with high levels of glycoside hydrolase activities. Near-complete genomes were reconstructed for the most abundant populations, which included composite genomes for populations closely related to sequenced strains of Thermus thermophilus and Rhodothermus marinus, and for novel populations that are related to thermophilicmore » Paenibacilli and an uncultivated subdivision of the littlestudied Gemmatimonadetes phylum. Partial genomes were also reconstructed for a number of lower abundance thermophilic Chloroflexi populations. Identification of genes for lignocellulose processing and metabolic reconstructions suggested Rhodothermus, Paenibacillus and Gemmatimonadetes as key groups for deconstructing biomass, and Thermus as a group that may primarily metabolize low molecular weight compounds. Mass spectrometry-based proteomic analysis of the consortium was used to identify .3000 proteins in fractionated samples from the cultures, and confirmed the importance of Paenibacillus and Gemmatimonadetes to biomass deconstruction. These studies also indicate that there are unexplored proteins with important roles in bacterial lignocellulose deconstruction.« less

  16. Physics and evolution of thermophilic adaptation.

    PubMed

    Berezovsky, Igor N; Shakhnovich, Eugene I

    2005-09-06

    Analysis of structures and sequences of several hyperthermostable proteins from various sources reveals two major physical mechanisms of their thermostabilization. The first mechanism is "structure-based," whereby some hyperthermostable proteins are significantly more compact than their mesophilic homologues, while no particular interaction type appears to cause stabilization; rather, a sheer number of interactions is responsible for thermostability. Other hyperthermostable proteins employ an alternative, "sequence-based" mechanism of their thermal stabilization. They do not show pronounced structural differences from mesophilic homologues. Rather, a small number of apparently strong interactions is responsible for high thermal stability of these proteins. High-throughput comparative analysis of structures and complete genomes of several hyperthermophilic archaea and bacteria revealed that organisms develop diverse strategies of thermophilic adaptation by using, to a varying degree, two fundamental physical mechanisms of thermostability. The choice of a particular strategy depends on the evolutionary history of an organism. Proteins from organisms that originated in an extreme environment, such as hyperthermophilic archaea (Pyrococcus furiosus), are significantly more compact and more hydrophobic than their mesophilic counterparts. Alternatively, organisms that evolved as mesophiles but later recolonized a hot environment (Thermotoga maritima) relied in their evolutionary strategy of thermophilic adaptation on "sequence-based" mechanism of thermostability. We propose an evolutionary explanation of these differences based on physical concepts of protein designability.

  17. Purification and characterization of 2-oxoglutarate:ferredoxin oxidoreductase from a thermophilic, obligately chemolithoautotrophic bacterium, Hydrogenobacter thermophilus TK-6.

    PubMed Central

    Yoon, K S; Ishii, M; Igarashi, Y; Kodama, T

    1996-01-01

    2-Oxoglutarate:ferredoxin oxidoreductase from a thermophilic, obligately autotrophic, hydrogen-oxidizing bacterium, Hydrogenobacter thermophilus TK-6, was purified to homogeneity by precipitation with ammonium sulfate and by fractionation by DEAE-Sepharose CL-6B, polyacrylate-quaternary amine, hydroxyapatite, and Superdex-200 chromatography. The purified enzyme had a molecular mass of about 105 kDa and comprised two subunits (70 kDa and 35 kDa). The activity of the 2-oxoglutarate:ferredoxin oxidoreductase was detected by the use of 2-oxoglutarate, coenzyme A, and one of several electron acceptors in substrate amounts (ferredoxin isolated from H. thermophilus, flavin adenine dinucleotide, flavin mononucleotide, or methyl viologen). NAD, NADP, and ferredoxins from Chlorella spp. and Clostridium pasteurianum were ineffective. The enzyme was extremely thermostable; the temperature optimum for 2-oxoglutarate oxidation was above 80 degrees C, and the time for a 50% loss of activity at 70 degrees C under anaerobic conditions was 22 h. The optimum pH for a 2-oxoglutarate oxidation reaction was 7.6 to 7.8. The apparent Km values for 2-oxoglutarate and coenzyme A at 70 degrees C were 1.42 mM and 80 microM, respectively. PMID:8655524

  18. Investigation of the thermophilic mechanism in the genus Porphyrobacter by comparative genomic analysis.

    PubMed

    Xu, Lin; Wu, Yue-Hong; Zhou, Peng; Cheng, Hong; Liu, Qian; Xu, Xue-Wei

    2018-05-23

    Type strains of the genus Porphyrobacter belonging to the family Erythrobacteraceae and the class Alphaproteobacteria have been isolated from various environments, such as swimming pools, lake water and hot springs. P. cryptus DSM 12079 T and P. tepidarius DSM 10594 T out of all Erythrobacteraceae type strains, are two type strains that have been isolated from geothermal environments. Next-generation sequencing (NGS) technology offers a convenient approach for detecting situational types based on protein sequence differences between thermophiles and mesophiles; amino acid substitutions can lead to protein structural changes, improving the thermal stabilities of proteins. Comparative genomic studies have revealed that different thermal types exist in different taxa, and few studies have been focused on the class Alphaproteobacteria, especially the family Erythrobacteraceae. In this study, eight genomes of Porphyrobacter strains were compared to elucidate how Porphyrobacter thermophiles developed mechanisms to adapt to thermal environments. P. cryptus DSM 12079 T grew optimally at 50 °C, which was higher than the optimal growth temperature of other Porphyrobacter type strains. Phylogenomic analysis of the genus Porphyrobacter revealed that P. cryptus DSM 12079 T formed a distinct and independent clade. Comparative genomic studies uncovered that 1405 single-copy genes were shared by Porphyrobacter type strains. Alignments of single-copy proteins showed that various types of amino acid substitutions existed between P. cryptus DSM 12079 T and the other Porphyrobacter strains. The primary substitution types were changes from glycine/serine to alanine. P. cryptus DSM 12079 T was the sole thermophile within the genus Porphyrobacter. Phylogenomic analysis and amino acid frequencies indicated that amino acid substitutions might play an important role in the thermophily of P. cryptus DSM 12079 T . Bioinformatic analysis revealed that major amino acid substitutional types

  19. Growth and metabolic profiling of the novel thermophilic bacterium Thermoanaerobacter sp. strain YS13.

    PubMed

    Peng, Tingting; Pan, Siyi; Christopher, Lew P; Sparling, Richard; Levin, David B

    2016-09-01

    A strictly anaerobic, thermophilic bacterium, designated strain YS13, was isolated from a geothermal hot spring. Phylogenetic analysis using the 16S rRNA genes and cpn60 UT genes suggested strain YS13 as a species of Thermoanaerobacter. Using cellobiose or xylose as carbon source, YS13 was able to grow over a wide range of temperatures (45-70 °C), and pHs (pH 5.0-9.0), with optimum growth at 65 °C and pH 7.0. Metabolic profiling on cellobiose, glucose, or xylose in 1191 medium showed that H2, CO2, ethanol, acetate, and lactate were the major metabolites. Lactate was the predominant end product from glucose or cellobiose fermentations, whereas H2 and acetate were the dominant end products from xylose fermentation. The metabolic balance shifted away from ethanol to H2, acetate, and lactate when YS13 was grown on cellobiose as temperatures increased from 45 to 70 °C. When YS13 was grown on xylose, a metabolic shift from lactate to H2, CO2, and acetate was observed in cultures as the temperature of incubation increased from 45 to 65 °C, whereas a shift from ethanol and CO2 to H2, acetate, and lactate was observed in cultures incubated at 70 °C.

  20. A Numerical Evaluation on the Viability of Heap Thermophilic Bioleaching of Chalcopyrite

    NASA Astrophysics Data System (ADS)

    Vilcaez, J.; Suto, K.; Inoue, C.

    2007-03-01

    The present numerical evaluation explores into the interactions among the many variables governing the mass and heat transport processes that take place in a heap thermophilic bioleaching system. The necessity of using mesophiles together with thermophiles is proved by tracing the activity of both microorganisms individually at each point throughout the heap. The role of key variables such as the fraction of FeS2 per CuFeS2 leached was quantified and its importance highlighted. In this evaluation, the heat transfer process plays the main role because of the heat accumulation required to maintain the heap temperature within the range of 60 °C to 80 °C where thermophilic microorganisms are capable of completing the unfinished dissolution of copper started by mesophilic microorganisms at 30 °C. The evaluation was done taking into consideration: biological activity as function of the temperature in the heap, heat loss due to conduction and advection from the top and bottom of the heap, and mass transfer between the gas and liquid phases as a function of temperature. The exothermic nature of the leaching reactions of CuFeS2 and FeS2 makes the system auto-thermal.

  1. Waste activated sludge hydrolysis and short-chain fatty acids accumulation under mesophilic and thermophilic conditions: effect of pH.

    PubMed

    Zhang, Peng; Chen, Yinguang; Zhou, Qi

    2009-08-01

    The effect of pH (4.0-11.0) on waste activated sludge (WAS) hydrolysis and short-chain fatty acids (SCFAs) accumulation under mesophilic and thermophilic conditions were investigated. The WAS hydrolysis increased markedly in thermophilic fermentation compared to mesophilic fermentation at any pH investigated. The hydrolysis at alkaline pHs (8.0-11.0) was greater than that at acidic pHs, but both of the acidic and alkaline hydrolysis was higher than that pH uncontrolled under either mesophilic or thermophilic conditions. No matter in mesophilic or thermophilic fermentation, the accumulation of SCFAs at alkaline pHs was greater than at acidic or uncontrolled pHs. The optimum SCFAs accumulation was 0.298g COD/g volatile suspended solids (VSS) with mesophilic fermentation, and 0.368 with thermophilic fermentation, which was observed respectively at pH 9.0 and fermentation time 5 d and pH 8.0 and time 9 d. The maximum SCFAs productions reported in this study were much greater than that in the literature. The analysis of the SCFAs composition showed that acetic acid was the prevalent acid in the accumulated SCFAs at any pH investigated under both temperatures, followed by propionic acid and n-valeric acid. Nevertheless, during the entire mesophilic and thermophilic fermentation the activity of methanogens was inhibited severely at acid or alkaline pHs, and the highest methane concentration was obtained at pH 7.0 in most cases. The studies of carbon mass balance showed that during WAS fermentation the reduction of VSS decreased with the increase of pH, and the thermophilic VSS reduction was greater than the mesophilic one. Further investigation indicated that most of the reduced VSS was converted to soluble protein and carbohydrate and SCFAs in two fermentations systems, while little formed methane and carbon dioxide.

  2. The Feasibility of Thermophilic Caldimonas manganoxidans as a Platform for Efficient PHB Production.

    PubMed

    Hsiao, Li-Jung; Lin, Ji-Hong; Sankatumvong, Pantitra; Wu, Tzong-Ming; Li, Si-Yu

    2016-11-01

    Recently, poly(3-hydroxybutyrate) (PHB) has been found in a few thermophilic strains where several advantages can be gained from running fermentation at high temperatures. Caldimonas manganoxidans, a thermophilic gram-negative bacterium, was investigated for the feasibility as a PHB-producing strain. It is suggested that the best fermentation strategy for achieving the highest PHB concentration of 5.4 ± 1.1 g/L (from 20 g/L glucose) in 24 h is to use the fermentation conditions that are favored for the bacterial growth, yet temperature and pH should be chosen at conditions that are favored for the PHB content. Besides, the above fermentation conditions produce PHB that has a high molecular weight of 1274 kDa with a low polydispersity index (PDI) of 1.45, where the highest Mw of PHB of 1399 kDa (PDI of 1.32) is obtained in this study. To the best knowledge of authors, C. manganoxidans has the best PHB productivity among the thermophiles and is comparable to those common PHB-producing mesophiles.

  3. Can anaerobic digestion of sugar beet pulp support the circular economy? a study of biogas and nutrient potential

    NASA Astrophysics Data System (ADS)

    Suhartini, S.; Heaven, S.; Banks, C. J.

    2018-03-01

    Anaerobic digestion (AD), known as a biological process without oxygen to convert complex organic materials into biogas, is capable of processing large tonnage quantities of biomass, such as sugar beet pulp (SBP). In addition to biogas production, its use allows nutrients and organic carbon recycle back to agriculture through the spreading of digestate. Digestate still contains high amount of nutrients (N, P, K) for use as biofertiliser. The aims of this research were to determine biogas/methane potential as a baseline for comparison with performance in semi-continuous digestion, and to determine nutrient and potentially toxic elements (PTE) of digestate fractions with respect to their potential for utilisation in agriculture. The Biochemical Methane Potential (BMP) test was performed in triplicate against blank and positive controls over a period of 28 days with gas measured at regular intervals. Semi-continuous AD of SBP was operated under mesophilic and thermophilic condition for 206 and 165 days. The results indicated that SBP is a very promising feedstock for AD, with the average BMP of 0.321 l CH4 g-1 VS and biogas potential of 0.605 l g-1 VS. Under semi-continuous operation, SBP also demonstrated positive results. Digestates from mesophilic and thermophilic AD of SBP contained useful quantities of N, P and K, with an acceptable Ni concentration in accordance to limits for PTE. These results suggest that digestate has the potential to be utilised on agricultural and arable land. This study illustrated the positive effects of applying AD to the achievement of economic savings and environmental-friendly performance.

  4. Aeration control of thermophilic aerobic digestion using fluorescence monitoring.

    PubMed

    Kim, Young-Kee; Oh, Byung-Keun

    2009-01-01

    The thermophilic aerobic digestion (TAD) process is recognized as an effective method for rapid waste activated sludge (WAS) degradation and the deactivation of pathogenic microorganisms. Yet, high energy costs due to heating and aeration have limited the commercialization of economical TAD processes. Previous research on autothermal thermophilic aerobic digestion (ATAD) has already reduced the heating cost. However, only a few studies have focused on reducing the aeration cost. Therefore, this study applied a two-step aeration control strategy to a fill-and-draw mode semicontinuous TAD process. The NADH-dependent fluorescence was monitored throughout the TAD experiment, and the aeration rate shifted according to the fluorescence intensity. As a result, the simple two-step aeration control operation achieved a 20.3% reduction in the total aeration, while maintaining an effective and stable operation. It is also expected that more savings can be achieved with a further reduction of the lower aeration rate or multisegmentation of the aeration rate.

  5. Isolation and polyphasic characterization of a novel hyper catalase producing thermophilic bacterium for the degradation of hydrogen peroxide.

    PubMed

    Sooch, Balwinder Singh; Kauldhar, Baljinder Singh; Puri, Munish

    2016-11-01

    A newly isolated microbial strain of thermophilic genus Geobacillus has been described with emphasis on polyphasic characterization and its application for degradation of hydrogen peroxide. The validation of this thermophilic strain of genus Geobacillus designated as BSS-7 has been demonstrated by polyphasic taxonomy approaches through its morphological, biochemical, fatty acid methyl ester profile and 16S rDNA sequencing. This thermophilic species of Geobacillus exhibited growth at broad pH and temperature ranges coupled with production of extraordinarily high quantities of intracellular catalase, the latter of which as yet not been reported in any member of this genus. The isolated thermophilic bacterial culture BSS-7 exhibited resistance against a variety of organic solvents. The immobilized whole cells of the bacterium successfully demonstrated the degradation of hydrogen peroxide (H2O2) in a packed bed reactor. This strain has potential application in various analytical and diagnostic methods in the form of biosensors and biomarkers in addition to applications in the textile, paper, food and pharmaceutical industries.

  6. Genus-Wide Assessment of Lignocellulose Utilization in the Extremely Thermophilic Genus Caldicellulosiruptor by Genomic, Pangenomic, and Metagenomic Analyses.

    PubMed

    Lee, Laura L; Blumer-Schuette, Sara E; Izquierdo, Javier A; Zurawski, Jeffrey V; Loder, Andrew J; Conway, Jonathan M; Elkins, James G; Podar, Mircea; Clum, Alicia; Jones, Piet C; Piatek, Marek J; Weighill, Deborah A; Jacobson, Daniel A; Adams, Michael W W; Kelly, Robert M

    2018-05-01

    Metagenomic data from Obsidian Pool (Yellowstone National Park, USA) and 13 genome sequences were used to reassess genus-wide biodiversity for the extremely thermophilic Caldicellulosiruptor The updated core genome contains 1,401 ortholog groups (average genome size for 13 species = 2,516 genes). The pangenome, which remains open with a revised total of 3,493 ortholog groups, encodes a variety of multidomain glycoside hydrolases (GHs). These include three cellulases with GH48 domains that are colocated in the glucan degradation locus (GDL) and are specific determinants for microcrystalline cellulose utilization. Three recently sequenced species, Caldicellulosiruptor sp. strain Rt8.B8 (renamed here Caldicellulosiruptor morganii ), Thermoanaerobacter cellulolyticus strain NA10 (renamed here Caldicellulosiruptor naganoensis ), and Caldicellulosiruptor sp. strain Wai35.B1 (renamed here Caldicellulosiruptor danielii ), degraded Avicel and lignocellulose (switchgrass). C. morganii was more efficient than Caldicellulosiruptor bescii in this regard and differed from the other 12 species examined, both based on genome content and organization and in the specific domain features of conserved GHs. Metagenomic analysis of lignocellulose-enriched samples from Obsidian Pool revealed limited new information on genus biodiversity. Enrichments yielded genomic signatures closely related to that of Caldicellulosiruptor obsidiansis , but there was also evidence for other thermophilic fermentative anaerobes ( Caldanaerobacter , Fervidobacterium , Caloramator , and Clostridium ). One enrichment, containing 89.8% Caldicellulosiruptor and 9.7% Caloramator , had a capacity for switchgrass solubilization comparable to that of C. bescii These results refine the known biodiversity of Caldicellulosiruptor and indicate that microcrystalline cellulose degradation at temperatures above 70°C, based on current information, is limited to certain members of this genus that produce GH48 domain

  7. Genus-wide assessment of lignocellulose utilization in the extremely thermophilic Caldicellulosiruptor by genomic, pan-genomic and metagenomic analysis

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Lee, Laura L.; Blumer-Schuette, Sara E.; Izquierdo, Javier A.

    Metagenomic data from Obsidian Pool (Yellowstone National Park, USA) and thirteen genome sequences were used to re-assess genus-wide biodiversity for the extremely thermophilicCaldicellulosiruptor. The updated core-genome contains 1,401 ortholog groups (average genome size for thirteen species = 2,516 genes). The pan-genome, which remains open with a revised total of 3,493 ortholog groups, encodes a variety of multi-domain glycoside hydrolases (GH). These include three cellulases with GH48 domains that are co-located in the Glucan Degradation Locus (GDL) and are specific determinants for microcrystalline cellulose utilization. Three recently sequenced species,Caldicellulosiruptorsp. str. Rt8.B8 (re-named hereCaldicellulosiruptor morganii),Thermoanaerobacter cellulolyticusstr. NA10 (re-named hereCaldicellulosiruptor naganoensisNA10), andCaldicellulosiruptorsp. str.more » Wai35.B1 (re-named hereCaldicellulosiruptor danielii) degraded Avicel and lignocellulose (switchgrass).C. morganiiwas more efficient thanC. besciiin this regard and differed from the other twelve species examined here, both based on genome content and organization and in the specific domain features of conserved GHs. Metagenomic analysis of lignocellulose-enriched samples from Obsidian Pool revealed limited new information on genus biodiversity. Enrichments yielded genomic signatures closely related toCaldicellulosiruptor obsidiansis, but there was also evidence for other thermophilic fermentative anaerobes (Caldanaerobacter,Fervidobacterium,Caloramator, andClostridium). One enrichment, containing 89.7%Caldicellulosiruptorand 9.7%Caloramator, had a capacity for switchgrass solubilization comparable toC. bescii. These results refine the known biodiversity ofCaldicellulosiruptorand indicate that microcrystalline cellulose degradation at temperatures above 70°C, based on current information, is limited to certain members of this genus that produce GH48 domain-containing enzymes. The genus

  8. Genus-wide assessment of lignocellulose utilization in the extremely thermophilic Caldicellulosiruptor by genomic, pan-genomic and metagenomic analysis

    DOE PAGES

    Lee, Laura L.; Blumer-Schuette, Sara E.; Izquierdo, Javier A.; ...

    2018-02-23

    Metagenomic data from Obsidian Pool (Yellowstone National Park, USA) and thirteen genome sequences were used to re-assess genus-wide biodiversity for the extremely thermophilicCaldicellulosiruptor. The updated core-genome contains 1,401 ortholog groups (average genome size for thirteen species = 2,516 genes). The pan-genome, which remains open with a revised total of 3,493 ortholog groups, encodes a variety of multi-domain glycoside hydrolases (GH). These include three cellulases with GH48 domains that are co-located in the Glucan Degradation Locus (GDL) and are specific determinants for microcrystalline cellulose utilization. Three recently sequenced species,Caldicellulosiruptorsp. str. Rt8.B8 (re-named hereCaldicellulosiruptor morganii),Thermoanaerobacter cellulolyticusstr. NA10 (re-named hereCaldicellulosiruptor naganoensisNA10), andCaldicellulosiruptorsp. str.more » Wai35.B1 (re-named hereCaldicellulosiruptor danielii) degraded Avicel and lignocellulose (switchgrass).C. morganiiwas more efficient thanC. besciiin this regard and differed from the other twelve species examined here, both based on genome content and organization and in the specific domain features of conserved GHs. Metagenomic analysis of lignocellulose-enriched samples from Obsidian Pool revealed limited new information on genus biodiversity. Enrichments yielded genomic signatures closely related toCaldicellulosiruptor obsidiansis, but there was also evidence for other thermophilic fermentative anaerobes (Caldanaerobacter,Fervidobacterium,Caloramator, andClostridium). One enrichment, containing 89.7%Caldicellulosiruptorand 9.7%Caloramator, had a capacity for switchgrass solubilization comparable toC. bescii. These results refine the known biodiversity ofCaldicellulosiruptorand indicate that microcrystalline cellulose degradation at temperatures above 70°C, based on current information, is limited to certain members of this genus that produce GH48 domain-containing enzymes. The genus

  9. A Rigidifying Salt-Bridge Favors the Activity of Thermophilic Enzyme at High Temperatures at the Expense of Low-Temperature Activity

    PubMed Central

    Lam, Sonia Y.; Yeung, Rachel C. Y.; Yu, Tsz-Ha; Sze, Kong-Hung; Wong, Kam-Bo

    2011-01-01

    Background Thermophilic enzymes are often less active than their mesophilic homologues at low temperatures. One hypothesis to explain this observation is that the extra stabilizing interactions increase the rigidity of thermophilic enzymes and hence reduce their activity. Here we employed a thermophilic acylphosphatase from Pyrococcus horikoshii and its homologous mesophilic acylphosphatase from human as a model to study how local rigidity of an active-site residue affects the enzymatic activity. Methods and Findings Acylphosphatases have a unique structural feature that its conserved active-site arginine residue forms a salt-bridge with the C-terminal carboxyl group only in thermophilic acylphosphatases, but not in mesophilic acylphosphatases. We perturbed the local rigidity of this active-site residue by removing the salt-bridge in the thermophilic acylphosphatase and by introducing the salt-bridge in the mesophilic homologue. The mutagenesis design was confirmed by x-ray crystallography. Removing the salt-bridge in the thermophilic enzyme lowered the activation energy that decreased the activation enthalpy and entropy. Conversely, the introduction of the salt-bridge to the mesophilic homologue increased the activation energy and resulted in increases in both activation enthalpy and entropy. Revealed by molecular dynamics simulations, the unrestrained arginine residue can populate more rotamer conformations, and the loss of this conformational freedom upon the formation of transition state justified the observed reduction in activation entropy. Conclusions Our results support the conclusion that restricting the active-site flexibility entropically favors the enzymatic activity at high temperatures. However, the accompanying enthalpy-entropy compensation leads to a stronger temperature-dependency of the enzymatic activity, which explains the less active nature of the thermophilic enzymes at low temperatures. PMID:21423654

  10. A rigidifying salt-bridge favors the activity of thermophilic enzyme at high temperatures at the expense of low-temperature activity.

    PubMed

    Lam, Sonia Y; Yeung, Rachel C Y; Yu, Tsz-Ha; Sze, Kong-Hung; Wong, Kam-Bo

    2011-03-01

    Thermophilic enzymes are often less active than their mesophilic homologues at low temperatures. One hypothesis to explain this observation is that the extra stabilizing interactions increase the rigidity of thermophilic enzymes and hence reduce their activity. Here we employed a thermophilic acylphosphatase from Pyrococcus horikoshii and its homologous mesophilic acylphosphatase from human as a model to study how local rigidity of an active-site residue affects the enzymatic activity. Acylphosphatases have a unique structural feature that its conserved active-site arginine residue forms a salt-bridge with the C-terminal carboxyl group only in thermophilic acylphosphatases, but not in mesophilic acylphosphatases. We perturbed the local rigidity of this active-site residue by removing the salt-bridge in the thermophilic acylphosphatase and by introducing the salt-bridge in the mesophilic homologue. The mutagenesis design was confirmed by x-ray crystallography. Removing the salt-bridge in the thermophilic enzyme lowered the activation energy that decreased the activation enthalpy and entropy. Conversely, the introduction of the salt-bridge to the mesophilic homologue increased the activation energy and resulted in increases in both activation enthalpy and entropy. Revealed by molecular dynamics simulations, the unrestrained arginine residue can populate more rotamer conformations, and the loss of this conformational freedom upon the formation of transition state justified the observed reduction in activation entropy. Our results support the conclusion that restricting the active-site flexibility entropically favors the enzymatic activity at high temperatures. However, the accompanying enthalpy-entropy compensation leads to a stronger temperature-dependency of the enzymatic activity, which explains the less active nature of the thermophilic enzymes at low temperatures.

  11. Wingate Anaerobic Test peak power and anaerobic capacity classifications for men and women intercollegiate athletes.

    PubMed

    Zupan, Michael F; Arata, Alan W; Dawson, Letitia H; Wile, Alfred L; Payn, Tamara L; Hannon, Megan E

    2009-12-01

    The Wingate Anaerobic Test (WAnT) has been established as an effective tool in measuring both muscular power and anaerobic capacity in a 30-second time period; however, there are no published normative tables by which to compare WAnT performance in men and women intercollegiate athletics. The purpose of this study was to develop a classification system for anaerobic peak power and anaerobic capacity for men and women National Collegiate Athletic Association (NCAA) Division I college athletes using the WAnT. A total of 1,585 (1,374 men and 211 women) tests were conducted on athletes ranging from the ages of 18 to 25 years using the WAnT. Absolute and relative peak power and anaerobic capacity data were recorded. One-half standard deviations were used to set up a 7-tier classification system (poor to elite) for these assessments. These classifications can be used by athletes, coaches, and practitioners to evaluate anaerobic peak power and anaerobic capacity in their athletes.

  12. Anaerobic digestion of stillage to produce bioenergy in the sugarcane-to-ethanol industry.

    PubMed

    Fuess, Lucas Tadeu; Garcia, Marcelo Loureiro

    2014-01-01

    Stillage is the main wastewater from ethanol production, containing a high chemical oxygen demand in addition to acidic and corrosive characteristics. Though stillage may be used as a soil fertilizer, its land application may be considered problematic due its high polluting potential. Anaerobic digestion represents an effective alternative treatment to reduce the pollution load of stillage. In addition, the methane gas produced within the process may be converted to energy, which can be directly applied to the treatment plant. The objective of this paper was to investigate the energetic potential of anaerobic digestion applied to stillage in the sugarcane ethanol industry. An overall analysis of the results indicates energy recovery capacity (ERC) values for methane ranging from 3.5% to 10%, respectively, for sugarcane juice and molasses. The processes employed to obtain the fermentable broth, as well as the distillation step, represent the main limiting factors to the energetic potential feasibility. Considering financial aspects the annual savings could reach up to US$ 30 million due to anaerobic digestion of stillage in relatively large-scale distilleries (365,000 m3 of ethanol per year). The best scenarios were verified for the association between anaerobic digestion of stillage and combustion of bagasse. In this case, the fossil fuels consumption in distilleries could be fully ceased, such the ERC of methane could reach values ranging from 140% to 890%.

  13. Aminomonas paucivorans gen. nov., sp. nov., a mesophilic, anaerobic, amino-acid-utilizing bacterium.

    PubMed

    Baena, S; Fardeau, M L; Ollivier, B; Labat, M; Thomas, P; Garcia, J L; Patel, B K

    1999-07-01

    A novel, asaccharolytic, amino-acid-degrading bacterium, designated strain GLU-3T, was isolated from an anaerobic lagoon of a dairy wastewater treatment plant. Strain GLU-3T stained Gram-negative and was an obligately anaerobic, non-spore-forming, slightly curved, rod-shaped bacterium (0.3 x 4.0-6.0 microns) which existed singly or in pairs. The DNA G+C content was 43 mol%. Optimum growth occurred at 35 degrees C and pH 7.5 on arginine with a generation time of 16 h. Good growth was obtained on arginine, histidine, threonine and glycine. Acetate was the end-product formed from all these substrates, but in addition, a trace of formate was detected from arginine and histidine, and ornithine was produced from arginine. Strain GLU-3T grew slowly on glutamate and produced acetate, carbon dioxide, formate, hydrogen and traces of propionate as the end-products. In syntrophic association with Methanobacterium formicicum, strain GLU-3T oxidized arginine, histidine and glutamate to give propionate as the major product; acetate, carbon dioxide and methane were also produced. Strain GLU-3T did not degrade alanine and the branched-chain amino acids valine, leucine and isoleucine either in pure culture or in association with M. formicicum. The nearest phylogenetic relative of strain GLU-3T was the thermophile Selenomonas acidaminovorans (similarity value of 89.5%). As strain GLU-3T is phylogenetically, physiologically and genotypically different from other amino-acid-degrading genera, it is proposed that it should be designated a new species of a new genus Aminomonas paucivorans gen. nov., sp. nov. (DSM 12260T).

  14. Thermophilic archaeal enzymes and applications in biocatalysis.

    PubMed

    Littlechild, Jennifer A

    2011-01-01

    Thermophilic enzymes have advantages for their use in commercial applications and particularly for the production of chiral compounds to produce optically pure pharmaceuticals. They can be used as biocatalysts in the application of 'green chemistry'. The thermophilic archaea contain enzymes that have already been used in commercial applications such as the L-aminoacylase from Thermococcus litoralis for the resolution of amino acids and amino acid analogues. This enzyme differs from bacterial L-aminoacylases and has similarities to carboxypeptidases from other archaeal species. An amidase/γ-lactamase from Sulfolobus solfataricus has been used for the production of optically pure γ-lactam, the building block for antiviral carbocyclic nucleotides. This enzyme has similarities to the bacterial signature amidase family. An alcohol dehydrogenase from Aeropyrum pernix has been used for the production of optically pure alcohols and is related to the zinc-containing eukaryotic alcohol dehydrogenases. A transaminase and a dehalogenase from Sulfolobus species have also been studied. The archaeal transaminase is found in a pathway for serine synthesis which is found only in eukaryotes and not in bacteria. It can be used for the asymmetric synthesis of homochiral amines of high enantioselective purity. The L-2-haloacid dehalogenase has applications both in biocatalysis and in bioremediation. All of these enzymes have increased thermostability over their mesophilic counterparts.

  15. Fermentation of sugarcane bagasse and chicken manure to calcium carboxylates under thermophilic conditions.

    PubMed

    Fu, Zhihong; Holtzapple, Mark T

    2010-09-01

    Sugarcane bagasse and chicken manure were anaerobically fermented to carboxylic acids using a mixed culture of marine microorganisms at 55 degrees C. Using the MixAlco process--an example of consolidated bioprocessing--the resulting carboxylate salts can be converted to mixed alcohol fuels or gasoline. To enhance digestibility, sugarcane bagasse was lime pretreated with 0.1 g Ca(OH)(2)/g dry biomass at 100 degrees C for 2 h. Four-stage countercurrent fermentation of 80% sugarcane bagasse/20% chicken manure was performed at various volatile solids (VS) loading rates and liquid residence times. Calcium carbonate was used as a buffer during fermentation. The highest acid productivity of 0.79 g/(L day) occurred at a total acid concentration of 21.5 g/L. The highest conversion (0.59 g VS digested/g VS fed) and yield (0.18 g total acids/g VS fed) occurred at a total acid concentration of 15.5 g/L. The continuum particle distribution model (CPDM) predicted the experimental total acid concentrations and conversions at an average error of 10.14% and 12.68%, respectively. CPDM optimizations show that high conversion (>80%) and total acid concentration of 21.3 g/L are possible with 300 g substrate/(L liquid), 30 days liquid residence time, and 3 g/(L day) solid loading rate. Thermophilic fermentation has a higher acetate content (approximately 63 wt%) than mesophilic fermentation (approximately 39 wt%).

  16. Microbial diversity of thermophiles with biomass deconstruction potential in a foliage-rich hot spring.

    PubMed

    Lee, Li Sin; Goh, Kian Mau; Chan, Chia Sing; Annie Tan, Geok Yuan; Yin, Wai-Fong; Chong, Chun Shiong; Chan, Kok-Gan

    2018-03-30

    The ability of thermophilic microorganisms and their enzymes to decompose biomass have attracted attention due to their quick reaction time, thermostability, and decreased risk of contamination. Exploitation of efficient thermostable glycoside hydrolases (GHs) could accelerate the industrialization of biofuels and biochemicals. However, the full spectrum of thermophiles and their enzymes that are important for biomass degradation at high temperatures have not yet been thoroughly studied. We examined a Malaysian Y-shaped Sungai Klah hot spring located within a wooded area. The fallen foliage that formed a thick layer of biomass bed under the heated water of the Y-shaped Sungai Klah hot spring was an ideal environment for the discovery and analysis of microbial biomass decay communities. We sequenced the hypervariable regions of bacterial and archaeal 16S rRNA genes using total community DNA extracted from the hot spring. Data suggested that 25 phyla, 58 classes, 110 orders, 171 families, and 328 genera inhabited this hot spring. Among the detected genera, members of Acidimicrobium, Aeropyrum, Caldilinea, Caldisphaera, Chloracidobacterium, Chloroflexus, Desulfurobacterium, Fervidobacterium, Geobacillus, Meiothermus, Melioribacter, Methanothermococcus, Methanotorris, Roseiflexus, Thermoanaerobacter, Thermoanaerobacterium, Thermoanaerobaculum, and Thermosipho were the main thermophiles containing various GHs that play an important role in cellulose and hemicellulose breakdown. Collectively, the results suggest that the microbial community in this hot spring represents a good source for isolating efficient biomass degrading thermophiles and thermozymes. © 2018 The Authors. MicrobiologyOpen published by John Wiley & Sons Ltd.

  17. Effect of heating strategy on power consumption and performance of a pilot plant anaerobic digester.

    PubMed

    Espinosa-Solares, Teodoro; Valle-Guadarrama, Salvador; Bombardiere, John; Domaschko, Max; Easter, Michael

    2009-05-01

    The effect of heating strategy on power consumption and performance of a pilot plant anaerobic digester treating chicken litter, under thermophilic conditions, has been studied. Heating strategy was evaluated using three different spans (0.2 degrees C, 0.6 degrees C, and 1.0 degree C) for triggering the temperature control system from target temperature (56.7 degrees C). The hydraulic retention time in the pilot plant digester was in the range of 32 to 37 days, varying the total solids concentration fed from 5% to 6%. The results showed that under the experimental conditions, heating was the most energy-demanding process with 95.5% of the energy used. Increments up to 7.5% and 3.8%, respectively, on mechanical and heating power consumption, were observed as the span, for triggering the temperature control system from target temperature, was increased. Under the experimental conditions studied here, an increment of 30.6% on the global biodigester performance index was observed when a span of 1.0 degree C was compared to the one of 0.2 degrees C.

  18. Cultivation of Anaerobic and Facultatively Anaerobic Bacteria from Spacecraft-Associated Clean Rooms▿

    PubMed Central

    Stieglmeier, Michaela; Wirth, Reinhard; Kminek, Gerhard; Moissl-Eichinger, Christine

    2009-01-01

    In the course of this biodiversity study, the cultivable microbial community of European spacecraft-associated clean rooms and the Herschel Space Observatory located therein were analyzed during routine assembly operations. Here, we focused on microorganisms capable of growing without oxygen. Anaerobes play a significant role in planetary protection considerations since extraterrestrial environments like Mars probably do not provide enough oxygen for fully aerobic microbial growth. A broad assortment of anaerobic media was used in our cultivation strategies, which focused on microorganisms with special metabolic skills. The majority of the isolated strains grew on anaerobic, complex, nutrient-rich media. Autotrophic microorganisms or microbes capable of fixing nitrogen were also cultivated. A broad range of facultatively anaerobic bacteria was detected during this study and also, for the first time, some strictly anaerobic bacteria (Clostridium and Propionibacterium) were isolated from spacecraft-associated clean rooms. The multiassay cultivation approach was the basis for the detection of several bacteria that had not been cultivated from these special environments before and also led to the discovery of two novel microbial species of Pseudomonas and Paenibacillus. PMID:19363082

  19. Cultivation of anaerobic and facultatively anaerobic bacteria from spacecraft-associated clean rooms.

    PubMed

    Stieglmeier, Michaela; Wirth, Reinhard; Kminek, Gerhard; Moissl-Eichinger, Christine

    2009-06-01

    In the course of this biodiversity study, the cultivable microbial community of European spacecraft-associated clean rooms and the Herschel Space Observatory located therein were analyzed during routine assembly operations. Here, we focused on microorganisms capable of growing without oxygen. Anaerobes play a significant role in planetary protection considerations since extraterrestrial environments like Mars probably do not provide enough oxygen for fully aerobic microbial growth. A broad assortment of anaerobic media was used in our cultivation strategies, which focused on microorganisms with special metabolic skills. The majority of the isolated strains grew on anaerobic, complex, nutrient-rich media. Autotrophic microorganisms or microbes capable of fixing nitrogen were also cultivated. A broad range of facultatively anaerobic bacteria was detected during this study and also, for the first time, some strictly anaerobic bacteria (Clostridium and Propionibacterium) were isolated from spacecraft-associated clean rooms. The multiassay cultivation approach was the basis for the detection of several bacteria that had not been cultivated from these special environments before and also led to the discovery of two novel microbial species of Pseudomonas and Paenibacillus.

  20. Development of a continuous bioconversion system using a thermophilic whole-cell biocatalyst.

    PubMed

    Ninh, Pham Huynh; Honda, Kohsuke; Yokohigashi, Yukako; Okano, Kenji; Omasa, Takeshi; Ohtake, Hisao

    2013-03-01

    The heat treatment of recombinant mesophilic cells having heterologous thermophilic enzymes results in the denaturation of indigenous mesophilic enzymes and the elimination of undesired side reactions; therefore, highly selective whole-cell catalysts comparable to purified enzymes can be readily prepared. However, the thermolysis of host cells leads to the heat-induced leakage of thermophilic enzymes, which are produced as soluble proteins, limiting the exploitation of their excellent stability in repeated and continuous reactions. In this study, Escherichia coli cells having the thermophilic fumarase from Thermus thermophilus (TtFTA) were treated with glutaraldehyde to prevent the heat-induced leakage of the enzyme, and the resulting cells were used as a whole-cell catalyst in repeated and continuous reactions. Interestingly, although electron microscopic observations revealed that the cellular structure of glutaraldehyde-treated E. coli was not apparently changed by the heat treatment, the membrane permeability of the heated cells to relatively small molecules (up to at least 3 kDa) was significantly improved. By applying the glutaraldehyde-treated E. coli having TtFTA to a continuous reactor equipped with a cell-separation membrane filter, the enzymatic hydration of fumarate to malate could be operated for more than 600 min with a molar conversion yield of 60% or higher.