WorldWideScience

Sample records for anaerobic enrichment culture

  1. Enrichment culture of marine anaerobic ammonium oxidation (anammox) bacteria

    Institute of Scientific and Technical Information of China (English)

    GUAN Yong-jie

    2016-01-01

    The present study investigates the enrichment of anaerobic ammonium oxidation (anammox) bacteria in the marine environment using sediment samples obtained from the East China Sea and discusses the nitrogen removal efficiency of marine anammox bioreactor. Enrichment of anammox bacteria with simultaneous removal of nitrite and ammonium ions was observed in the Anaerobic Sequencing Batch Reactor under a total nitrogen loading rate of 0.37kg-N m-3day-1. In this study, The nitrogen removal efficiency was up to 80% and the molar-reaction ratio of ammonium, nitrite and nitrate was 1.0:1.22:0.22 which was a little different from a previously reported ratio of 1.0:1.32:0.26 in a freshwater system.

  2. Microbial dynamics in anaerobic enrichment cultures degrading di-n-butyl phthalic acid ester

    DEFF Research Database (Denmark)

    Trably, Eric; Batstone, Damien J.; Christensen, Nina

    2008-01-01

    in enrichment cultures degrading phthalic acid esters under methanogenic conditions. A selection pressure was applied by adding DBP at 10 and 200 mg L(-1) in semi-continuous anaerobic reactors. The microbial dynamics were monitored using single strand conformation polymorphism (SSCP). While only limited abiotic...

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

    DEFF Research Database (Denmark)

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

    2009-01-01

    The aim of this study was to enrich, characterize and identify strict anaerobic extreme thermophilic hydrogen (H-2) producers from digested household solid wastes. A strict anaerobic extreme thermophilic H-2 producing bacterial culture was enriched from a lab-scale digester treating household was...... from digested household wastes. This study provided a culture with a potential to be applied in reactor systems for extreme thermophilic H-2 production from complex organic wastes.......The aim of this study was to enrich, characterize and identify strict anaerobic extreme thermophilic hydrogen (H-2) producers from digested household solid wastes. A strict anaerobic extreme thermophilic H-2 producing bacterial culture was enriched from a lab-scale digester treating household...... sources. Growth on glucose produced acetate, H-2 and carbon dioxide. Maximal H-2 production rate on glucose was 1.1 mmol l(-1) h(-1) with a maximum H-2 yield of 1.9 mole H-2 per mole glucose. 16S ribosomal DNA clone library analyses showed that the culture members were phylogenetically affiliated...

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

    DEFF Research Database (Denmark)

    Luo, Gang; Angelidaki, Irini

    2012-01-01

    the existing natural gas grid. The current study presents a new biological method for biogas upgrading in a separate biogas reactor, containing enriched hydrogenotrophic methanogens and fed with biogas and hydrogen. Both mesophilic- and thermophilic anaerobic cultures were enriched to convert CO2 to CH4...... by 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......Biogas produced by anaerobic digestion, is mainly used in a gas motor for heat and electricity production. However, after removal of CO2, biogas can be upgraded to natural gas quality, giving more utilization possibilities, such as utilization as autogas, or distant utilization by using...

  5. Use of {gamma}-hexachlorocyclohexane as a terminal electron acceptor by an anaerobic enrichment culture

    Energy Technology Data Exchange (ETDEWEB)

    Elango, Vijai, E-mail: velango@g.clemson.edu [Hazardous Substance Research Center/South and Southwest, Louisiana State University, 3221 Patrick Taylor Hall, Baton Rouge, LA 70803 (United States); Kurtz, Harry D. [Department of Genetics and Biochemistry, Clemson University, 100 Jordan Hall, Clemson, SC 29634 (United States); Anderson, Christina; Freedman, David L. [Department of Environmental Engineering and Earth Sciences, Box 340919, Clemson University, Clemson, SC 29634-0919 (United States)

    2011-12-15

    Highlights: Black-Right-Pointing-Pointer Use of {gamma}-hexachlorocyclohexane as a terminal electron acceptor was demonstrated. Black-Right-Pointing-Pointer H{sub 2} served as the electron donor for an enrichment culture that dechlorinated {gamma}-HCH. Black-Right-Pointing-Pointer H{sub 2} consumption for acetogenesis and methanogenesis stopped in HEPES media. Black-Right-Pointing-Pointer Addition of vancomycin significantly slowed the rate of {gamma}-HCH dechlorination. Black-Right-Pointing-Pointer Previously identified chlororespiring microbes were not detected in the enrichment. - Abstract: The use of {gamma}-hexachlorocyclohexane (HCH) as a terminal electron acceptor via organohalide respiration was demonstrated for the first time with an enrichment culture grown in a sulfate-free HEPES-buffered anaerobic mineral salts medium. The enrichment culture was initially developed with soil and groundwater from an industrial site contaminated with HCH isomers, chlorinated benzenes, and chlorinated ethenes. When hydrogen served as the electron donor, 79-90% of the electron equivalents from hydrogen were used by the enrichment culture for reductive dechlorination of the {gamma}-HCH, which was provided at a saturation concentration of approximately 10 mg/L. Benzene and chlorobenzene were the only volatile transformation products detected, accounting for 25% and 75% of the {gamma}-HCH consumed (on a molar basis), respectively. The enrichment culture remained active with only hydrogen as the electron donor and {gamma}-HCH as the electron acceptor through several transfers to fresh mineral salts medium for more than one year. Addition of vancomycin to the culture significantly slowed the rate of {gamma}-HCH dechlorination, suggesting that a Gram-positive organism is responsible for the reduction of {gamma}-HCH. Analysis of the {gamma}-HCH dechlorinating enrichment culture did not detect any known chlororespiring genera, including Dehalobacter. In bicarbonate-buffered medium

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

    Science.gov (United States)

    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).

  7. Identification of enzymes involved in anaerobic benzene degradation by a strictly anaerobic iron-reducing enrichment culture.

    Science.gov (United States)

    Abu Laban, Nidal; Selesi, Draženka; Rattei, Thomas; Tischler, Patrick; Meckenstock, Rainer U

    2010-10-01

    Anaerobic benzene degradation was studied with a highly enriched iron-reducing culture (BF) composed of mainly Peptococcaceae-related Gram-positive microorganisms. The proteomes of benzene-, phenol- and benzoate-grown cells of culture BF were compared by SDS-PAGE. A specific benzene-expressed protein band of 60 kDa, which could not be observed during growth on phenol or benzoate, was subjected to N-terminal sequence analysis. The first 31 amino acids revealed that the protein was encoded by ORF 138 in the shotgun sequenced metagenome of culture BF. ORF 138 showed 43% sequence identity to phenylphosphate carboxylase subunit PpcA of Aromatoleum aromaticum strain EbN1. A LC/ESI-MS/MS-based shotgun proteomic analysis revealed other specifically benzene-expressed proteins with encoding genes located adjacent to ORF 138 on the metagenome. The protein products of ORF 137, ORF 139 and ORF 140 showed sequence identities of 37% to phenylphosphate carboxylase PpcD of A. aromaticum strain EbN1, 56% to benzoate-CoA ligase (BamY) of Geobacter metallireducens and 67% to 3-octaprenyl-4-hydroxybenzoate carboxy-lyase (UbiD/UbiX) of A. aromaticum strain EbN1 respectively. These genes are proposed as constituents of a putative benzene degradation gene cluster (∼ 17 kb) composed of carboxylase-related genes. The identified gene sequences suggest that the initial activation reaction in anaerobic benzene degradation is probably a direct carboxylation of benzene to benzoate catalysed by putative anaerobic benzene carboxylase (Abc). The putative Abc probably consists of several subunits, two of which are encoded by ORFs 137 and 138, and belongs to a family of carboxylases including phenylphosphate carboxylase (Ppc) and 3-octaprenyl-4-hydroxybenzoate carboxy-lyase (UbiD/UbiX).

  8. High-efficiency hydrogen production by an anaerobic, thermophilic enrichment culture from an Icelandic hot spring.

    Science.gov (United States)

    Koskinen, Perttu E P; Lay, Chyi-How; Puhakka, Jaakko A; Lin, Ping-Jei; Wu, Shu-Yii; Orlygsson, Jóhann; Lin, Chiu-Yue

    2008-11-01

    Dark fermentative hydrogen production from glucose by a thermophilic culture (33HL), enriched from an Icelandic hot spring sediment sample, was studied in two continuous-flow, completely stirred tank reactors (CSTR1, CSTR2) and in one semi-continuous, anaerobic sequencing batch reactor (ASBR) at 58 degrees C. The 33HL produced H2 yield (HY) of up to 3.2 mol-H2/mol-glucose along with acetate in batch assay. In the CSTR1 with 33HL inoculum, H2 production was unstable. In the ASBR, maintained with 33HL, the H2 production enhanced after the addition of 6 mg/L of FeSO4 x H2O resulting in HY up to 2.51 mol-H2/mol-glucose (H2 production rate (HPR) of 7.85 mmol/h/L). The H2 production increase was associated with an increase in butyrate production. In the CSTR2, with ASBR inoculum and FeSO4 supplementation, stable, high-rate H2 production was obtained with HPR up to 45.8 mmol/h/L (1.1 L/h/L) and HY of 1.54 mol-H2/mol-glucose. The 33HL batch enrichment was dominated by bacterial strains closely affiliated with Thermobrachium celere (99.8-100%). T. celere affiliated strains, however, did not thrive in the three open system bioreactors. Instead, Thermoanaerobacterium aotearoense (98.5-99.6%) affiliated strains, producing H2 along with butyrate and acetate, dominated the reactor cultures. This culture had higher H2 production efficiency (HY and specific HPR) than reported for mesophilic mixed cultures. Further, the thermophilic culture readily formed granules in CSTR and ASBR systems. In summary, the thermophilic culture as characterized by high H2 production efficiency and ready granulation is considered very promising for H2 fermentation from carbohydrates.

  9. Inhibition of anaerobic ammonium oxidizing (anammox) enrichment cultures by substrates, metabolites and common wastewater constituents.

    Science.gov (United States)

    Carvajal-Arroyo, José M; Sun, Wenjie; Sierra-Alvarez, Reyes; Field, Jim A

    2013-03-01

    Anaerobic ammonium oxidation (anammox) is an emerging technology for nitrogen removal that provides a more environmentally sustainable and cost effective alternative compared to conventional biological treatment methods. The objective of this study was to investigate the inhibitory impact of anammox substrates, metabolites and common wastewater constituents on the microbial activity of two different anammox enrichment cultures (suspended and granular), both dominated by bacteria from the genus Brocadia. Inhibition was evaluated in batch assays by comparing the N(2) production rates in the absence or presence of each compound supplied in a range of concentrations. The optimal pH was 7.5 and 7.3 for the suspended and granular enrichment cultures, respectively. Among the substrates or products, ammonium and nitrate caused low to moderate inhibition, whereas nitrite caused almost complete inhibition at concentrations higher than 15 mM. The intermediate, hydrazine, either stimulated or caused low inhibition of anammox activity up to 3mM. Of the common constituents in wastewater, hydrogen sulfide was the most severe inhibitor, with 50% inhibitory concentrations (IC(50)) as low as 0.03 mM undissociated H(2)S. Dissolved O(2) showed moderate inhibition (IC(50)=2.3-3.8 mg L(-1)). In contrast, phosphate and salinity (NaCl) posed very low inhibition. The suspended- and granular anammox enrichment cultures had similar patterns of response to the various inhibitory stresses with the exception of phosphate. The findings of this study provide comprehensive insights on the tolerance of the anammox process to a wide variety of potential inhibiting compounds.

  10. Phylogenetic analysis of anaerobic psychrophilic enrichment cultures obtained from a greenland glacier ice core

    Science.gov (United States)

    Sheridan, Peter P.; Miteva, Vanya I.; Brenchley, Jean E.

    2003-01-01

    The examination of microorganisms in glacial ice cores allows the phylogenetic relationships of organisms frozen for thousands of years to be compared with those of current isolates. We developed a method for aseptically sampling a sediment-containing portion of a Greenland ice core that had remained at -9 degrees C for over 100,000 years. Epifluorescence microscopy and flow cytometry results showed that the ice sample contained over 6 x 10(7) cells/ml. Anaerobic enrichment cultures inoculated with melted ice were grown and maintained at -2 degrees C. Genomic DNA extracted from these enrichments was used for the PCR amplification of 16S rRNA genes with bacterial and archaeal primers and the preparation of clone libraries. Approximately 60 bacterial inserts were screened by restriction endonuclease analysis and grouped into 27 unique restriction fragment length polymorphism types, and 24 representative sequences were compared phylogenetically. Diverse sequences representing major phylogenetic groups including alpha, beta, and gamma Proteobacteria as well as relatives of the Thermus, Bacteroides, Eubacterium, and Clostridium groups were found. Sixteen clone sequences were closely related to those from known organisms, with four possibly representing new species. Seven sequences may reflect new genera and were most closely related to sequences obtained only by PCR amplification. One sequence was over 12% distant from its closest relative and may represent a novel order or family. These results show that phylogenetically diverse microorganisms have remained viable within the Greenland ice core for at least 100,000 years.

  11. Ammonia tolerant enriched methanogenic cultures as bioaugmentation inocula to alleviate ammonia inhibition in continuous anaerobic reactors

    DEFF Research Database (Denmark)

    Fotidis, Ioannis; Wang, Han; Angelidaki, Irini

    tolerant methanogenic culture as potential bioaugmentation inoculum in a continuous stirred tank reactor (CSTR) operating under “inhibited steady-state”, triggered by high ammonia levels (5 g NH4+-N L-1). The results of the current study established for the first time that bioaugmentation of an enriched...... ammonia tolerant methanogen in a CSTR reactor could completely alleviate the ammonia inhibitory effect. Furthermore, it was found that bioaugmentation with the enriched culture resulted in 25% higher methane production compared to when the bioaugmentation was achieved with pure methanogenic strains....... The bioaugmentation was performed without pausing the continuous operation of the CSTR reactor and without excluding the ammonia-rich substrate from the feedstock. Thus, bioaugmentation with mixed methanogenic cultures could potentially support the development of an efficient and cost-effective biomethanation process...

  12. Quantification of anaerobic ammonium-oxidizing bacteria in enrichment cultures by quantitative competitive PCR

    Institute of Scientific and Technical Information of China (English)

    HAO Chun; WANG Huan; LIU Qinhua; LI Xudong

    2009-01-01

    The anaerobic ammonium-oxidizing (ANAMMOX) bacteria were enriched from a sequencing batch biofilm reactor (SBBR) biofilm.We successfully developed a quantitative competitive polymerase chain reaction (QC-PCR) system to detect and quantify ANAMMOX bacteria in environmental samples.For QC-PCR system,PCR primer sets targeting 16S ribosomal RNA genes of ANAMMOX bacteria were designed and used.The quantification range of this system was 4 orders of magnitude,from 10~3 to 10~6 copies per PCR,corresponding to the detection limit of 300 target copies per mL.A 312-bp internal standard (IS) was constructed,which showed very similar amplification efficiency with the target amxC fragment (349 bp) over 4 orders of magnitude (10~3-10~6).The linear regressions were obtained with a R~2 of 0.9824 for 10~3 copies,R~2 of 0.9882 for 10~4 copies,0.9857 for 10~5 copies and 0.9899 for 10~6 copies.Using this method,we quantified ANAMMOX bacteria in a shortcut nitrification/denitrification-anammox system which is set for piggery wastewater treatment.

  13. Ultrastructure and viral metagenome of bacteriophages from an anaerobic methane oxidizing methylomirabilis bioreactor enrichment culture

    NARCIS (Netherlands)

    Gambelli, Lavinia; Cremers, Geert; Mesman, Rob; Guerrero, Simon; Dutilh, Bas E.; Jetten, Mike S M; den Camp, Huub J M Op; van Niftrik, Laura

    2016-01-01

    With its capacity for anaerobic methane oxidation and denitrification, the bacterium Methylomirabilis oxyfera plays an important role in natural ecosystems. Its unique physiology can be exploited for more sustainable wastewater treatment technologies. However, operational stability of full-scale bio

  14. Ammonia tolerant enriched methanogenic cultures as bioaugmentation inocula to alleviate ammonia inhibition in continuous anaerobic reactors

    DEFF Research Database (Denmark)

    Fotidis, Ioannis; Wang, Han; Angelidaki, Irini

    Ammonia is the most common inhibitor of anaerobic digestion (AD) process, resulting in suboptimal exploitation of the biogas potential of the feedstocks, causing significant economic losses to the biogas plants. Ammonia is mainly inhibiting the aceticlastic methanogens, while the hydrogenotrophic...... of ammonia-rich organic waste in full-scale continuous reactors.......Ammonia is the most common inhibitor of anaerobic digestion (AD) process, resulting in suboptimal exploitation of the biogas potential of the feedstocks, causing significant economic losses to the biogas plants. Ammonia is mainly inhibiting the aceticlastic methanogens, while the hydrogenotrophic...... methanogens are more robust to ammonia toxicity effect. It has been shown that bioaugmentation of a pure strain of a hydrogenotrophic methanogen (i.e. Methanoculleus bourgensis) in an ammonia inhibited continuous anaerobic reactor can improve methane production more than 30%. Nevertheless, cultivation...

  15. Establishment and Characterization of an Anaerobic Thermophilic (55 degrees C) Enrichment Culture Degrading Long-Chain Fatty Acids

    DEFF Research Database (Denmark)

    Angelidaki, Irini; Ahring, Birgitte Kiær

    1995-01-01

    A thermophilic, long-chain fatty acid-oxidizing culture was enriched. Stearate was used as the substrate, and methane and carbon dioxide were the sole end products. Cultivation was possible only when a fed-batch system was used or with addition of activated carbon or bentonite. The enrichment...

  16. Anaerobic alkane biodegradation by cultures enriched from oil sands tailings ponds involves multiple species capable of fumarate addition.

    Science.gov (United States)

    Tan, BoonFei; Semple, Kathleen; Foght, Julia

    2015-05-01

    A methanogenic short-chain alkane-degrading culture (SCADC) was enriched from oil sands tailings and transferred several times with a mixture of C6, C7, C8 and C10 n-alkanes as the predominant organic carbon source, plus 2-methylpentane, 3-methylpentane and methylcyclopentane as minor components. Cultures produced ∼40% of the maximum theoretical methane during 18 months incubation while depleting the n-alkanes, 2-methylpentane and methylcyclopentane. Substrate depletion correlated with detection of metabolites characteristic of fumarate activation of 2-methylpentane and methylcyclopentane, but not n-alkane metabolites. During active methanogenesis with the mixed alkanes, reverse-transcription PCR confirmed the expression of functional genes (assA and bssA) associated with hydrocarbon addition to fumarate. Pyrosequencing of 16S rRNA genes amplified during active alkane degradation revealed enrichment of Clostridia (particularly Peptococcaceae) and methanogenic Archaea (Methanosaetaceae and Methanomicrobiaceae). Methanogenic cultures transferred into medium containing sulphate produced sulphide, depleted n-alkanes and produced the corresponding succinylated alkane metabolites, but were slow to degrade 2-methylpentane and methylcyclopentane; these cultures were enriched in Deltaproteobacteria rather than Clostridia. 3-Methylpentane was not degraded by any cultures. Thus, nominally methanogenic oil sands tailings harbour dynamic and versatile hydrocarbon-degrading fermentative syntrophs and sulphate reducers capable of degrading n-, iso- and cyclo-alkanes by addition to fumarate.

  17. Anaerobic degradation of benzene by enriched consortia with humic acids as terminal electron acceptors

    Energy Technology Data Exchange (ETDEWEB)

    Cervantes, Francisco J., E-mail: fjcervantes@ipicyt.edu.mx [Division de Ciencias Ambientales, Instituto Potosino de Investigacion Cientifica y Tecnologica (IPICyT), Camino a la Presa San Jose 2055, Col. Lomas 4a. Seccion, San Luis Potosi, SLP, 78216 Mexico (Mexico); Mancilla, Ana Rosa; Toro, E. Emilia Rios-del [Division de Ciencias Ambientales, Instituto Potosino de Investigacion Cientifica y Tecnologica (IPICyT), Camino a la Presa San Jose 2055, Col. Lomas 4a. Seccion, San Luis Potosi, SLP, 78216 Mexico (Mexico); Alpuche-Solis, Angel G.; Montoya-Lorenzana, Lilia [Division de Biologia Molecular, Instituto Potosino de Investigacion Cientifica y Tecnologica (IPICyT), Camino a la Presa San Jose 2055, Col. Lomas 4a. Seccion, San Luis Potosi, SLP, 78216 Mexico (Mexico)

    2011-11-15

    Highlights: {yields} Enriched consortia were able to couple the anaerobic degradation of benzene to the reduction of humic acids. {yields} Electron-equivalents derived from anaerobic benzene oxidation were highly recovered as reduced humic acids. {yields} Several species from classes {beta}-, {delta}- and {gamma}-Proteobacteria were enriched during the anaerobic degradation of benzene. - Abstract: The anaerobic degradation of benzene coupled to the reduction of humic acids (HA) was demonstrated in two enriched consortia. Both inocula were able to oxidize benzene under strict anaerobic conditions when the humic model compound, anthraquinone-2,6-disulfonate (AQDS), was supplied as terminal electron acceptor. An enrichment culture originated from a contaminated soil was also able to oxidize benzene linked to the reduction of highly purified soil humic acids (HPSHA). In HPSHA-amended cultures, 9.3 {mu}M of benzene were degraded, which corresponds to 279 {+-} 27 micro-electron equivalents ({mu}Eq) L{sup -1}, linked to the reduction of 619 {+-} 81 {mu}Eq L{sup -1} of HPSHA. Neither anaerobic benzene oxidation nor reduction of HPSHA occurred in sterilized controls. Anaerobic benzene oxidation did not occur in soil incubations lacking HPSHA. Furthermore, negligible reduction of HPSHA occurred in the absence of benzene. The enrichment culture derived from this soil was dominated by two {gamma}-Proteobacteria phylotypes. A benzene-degrading AQDS-reducing enrichment originated from a sediment sample showed the prevalence of different species from classes {beta}-, {delta}- and {gamma}-Proteobacteria. The present study provides clear quantitative demonstration of anaerobic degradation of benzene coupled to the reduction of HA.

  18. Phylogenetic and metabolic diversity of hexahydro-1,3,5-trinitro-1,3,5-triazine (RDX)-transforming bacteria in strictly anaerobic mixed cultures enriched on RDX as nitrogen source.

    Science.gov (United States)

    Zhao, Jian-Shen; Spain, Jim; Hawari, Jalal

    2003-11-01

    Five obligate anaerobes that were most closely related to Clostridium bifermentans, Clostridium celerecrescens, Clostridium saccharolyticum, Clostridium butyricum and Desulfovibrio desulfuricans by their 16S rRNA genes sequences were isolated from enrichment cultures using hexahydro-1,3,5-trinitro-1,3,5-triazine (RDX) as a nitrogen source. The above isolates transformed RDX at rates of 24.0, 5.4, 6.2, 2.5, 5.5 mumol h(-1) g (dry weight) of cells(-1), respectively, to nitrite, formaldehyde, methanol, and nitrous oxide. The present results indicate that clostridia are major strains responsible for RDX removal, and all isolates seemed to mainly transform RDX via its initial reduction to MNX and subsequent denitration. Since clostridia are commonly present in soil, we suggest that they may contribute to the removal of RDX in the subsurface (anoxic) soil.

  19. Simple and convenient method for culturing anaerobic bacteria.

    OpenAIRE

    Behbehani, M J; Jordan, H. V.; Santoro, D L

    1982-01-01

    A simple and convenient method for culturing anaerobic bacteria is described. Cultures can be grown in commercially available flasks normally used for preparation of sterile external solutions. A special disposable rubber flask closure maintains anaerobic conditions in the flask after autoclaving. Growth of a variety of anaerobic oral bacteria was comparable to that obtained after anaerobic incubation of broth cultures in Brewer Anaerobic Jars.

  20. Enrichment of high ammonia tolerant methanogenic culture

    DEFF Research Database (Denmark)

    Fotidis, Ioannis; Karakashev, Dimitar Borisov; Proietti, Nicolas

    Ammonia is the major toxicant in full scale anaerobic digesters of animal wastes which are rich in proteins and/or urea, such as pig or poultry wastes. Ammonia inhibition decreases methane production rates, increases volatile fatty acids concentration and leads to economic losses for the biogas...... plants. The methods used today to counteract ammonia inhibition are slow and costexpensive. A new biological approach to avoid or counteract ammonia inhibition by using ammonia tolerant methanogens, could provide a sustainable solution for cost-effective digestion of abundant ammonia-rich wastes. The aim...... of the current study was to isolate and identify methanogenic cultures tolerant to high ammonia concentrations. A mixed methanogenic population was stepwise exposed to ammonia concentrations (1 to 9.26 g NH4+-N L-1) during an enrichment process with successive batch cultivations. The methanogenic population...

  1. Phylogenetic and Metabolic Diversity of Hexahydro-1,3,5-trinitro-1,3,5-triazine (RDX)-transforming Bacteria in Strictly Anaerobic Mixed Cultures Enriched on RDX as Nitrogen Source

    Science.gov (United States)

    2003-01-01

    the Enterobacteriaceae family ( Klebsiella pneumoniae, Serratia marcescens, Morganella morganii, Citrobacter freundii, and Escherichia coli) [10,11... wastewater to methane (70% of the total gas released). The following compounds were added to the basic salts and vitamins medium to enrich bacteria using RDX...RDX biodegradation by a methanogenic enrichment culture obtained from an explosives man- ufacturing wastewater treatment plant. Technical report, pp. 99

  2. Carbohydrate-enriched cyanobacterial biomass as feedstock for bio-methane production through anaerobic digestion

    DEFF Research Database (Denmark)

    Markou, Giorgos; Angelidaki, Irini; Georgakakis, Dimitris

    2013-01-01

    The anaerobic digestion performance using carbohydrate-enriched biomass of Arthrospira platensis was studied. The carbohydrate enrichment was achieved after the cultivation of A. platensis under phosphorus limitation conditions. Three biomass compositions (60%, 40% and 20% carbohydrates content) ...

  3. Diversity and enrichment of nitrite-dependent anaerobic methane oxidizing bacteria from wastewater sludge.

    Science.gov (United States)

    Luesken, Francisca A; van Alen, Theo A; van der Biezen, Erwin; Frijters, Carla; Toonen, Ger; Kampman, Christel; Hendrickx, Tim L G; Zeeman, Grietje; Temmink, Hardy; Strous, Marc; Op den Camp, Huub J M; Jetten, Mike S M

    2011-11-01

    Recently discovered microorganisms affiliated to the bacterial phylum NC10, named "Candidatus Methylomirabilis oxyfera", perform nitrite-dependent anaerobic methane oxidation. These microorganisms could be important players in a novel way of anaerobic wastewater treatment where ammonium and residual dissolved methane might be removed at the expense of nitrate or nitrite. To find suitable inocula for reactor startup, ten selected wastewater treatment plants (WWTPs) located in The Netherlands were screened for the endogenous presence of M. oxyfera using molecular diagnostic methods. We could identify NC10 bacteria with 98% similarity to M. oxyfera in nine out of ten WWTPs tested. Sludge from one selected WWTP was used to start a new enrichment culture of NC10 bacteria. This enrichment was monitored using specific pmoA primers and M. oxyfera cells were visualized with fluorescence oligonucleotide probes. After 112 days, the enrichment consumed up to 0.4 mM NO(2)(-) per day. The results of this study show that appropriate sources of biomass, enrichment strategies, and diagnostic tools existed to start and monitor pilot scale tests for the implementation of nitrite-dependent methane oxidation in wastewater treatment at ambient temperature.

  4. Culture enriched molecular profiling of the cystic fibrosis airway microbiome.

    Science.gov (United States)

    Sibley, Christopher D; Grinwis, Margot E; Field, Tyler R; Eshaghurshan, Christina S; Faria, Monica M; Dowd, Scot E; Parkins, Michael D; Rabin, Harvey R; Surette, Michael G

    2011-01-01

    The microbiome of the respiratory tract, including the nasopharyngeal and oropharyngeal microbiota, is a dynamic community of microorganisms that is highly diverse. The cystic fibrosis (CF) airway microbiome refers to the polymicrobial communities present in the lower airways of CF patients. It is comprised of chronic opportunistic pathogens (such as Pseudomonas aeruginosa) and a variety of organisms derived mostly from the normal microbiota of the upper respiratory tract. The complexity of these communities has been inferred primarily from culture independent molecular profiling. As with most microbial communities it is generally assumed that most of the organisms present are not readily cultured. Our culture collection generated using more extensive cultivation approaches, reveals a more complex microbial community than that obtained by conventional CF culture methods. To directly evaluate the cultivability of the airway microbiome, we examined six samples in depth using culture-enriched molecular profiling which combines culture-based methods with the molecular profiling methods of terminal restriction fragment length polymorphisms and 16S rRNA gene sequencing. We demonstrate that combining culture-dependent and culture-independent approaches enhances the sensitivity of either approach alone. Our techniques were able to cultivate 43 of the 48 families detected by deep sequencing; the five families recovered solely by culture-independent approaches were all present at very low abundance (<0.002% total reads). 46% of the molecular signatures detected by culture from the six patients were only identified in an anaerobic environment, suggesting that a large proportion of the cultured airway community is composed of obligate anaerobes. Most significantly, using 20 growth conditions per specimen, half of which included anaerobic cultivation and extended incubation times we demonstrate that the majority of bacteria present can be cultured.

  5. Culture enriched molecular profiling of the cystic fibrosis airway microbiome.

    Directory of Open Access Journals (Sweden)

    Christopher D Sibley

    Full Text Available The microbiome of the respiratory tract, including the nasopharyngeal and oropharyngeal microbiota, is a dynamic community of microorganisms that is highly diverse. The cystic fibrosis (CF airway microbiome refers to the polymicrobial communities present in the lower airways of CF patients. It is comprised of chronic opportunistic pathogens (such as Pseudomonas aeruginosa and a variety of organisms derived mostly from the normal microbiota of the upper respiratory tract. The complexity of these communities has been inferred primarily from culture independent molecular profiling. As with most microbial communities it is generally assumed that most of the organisms present are not readily cultured. Our culture collection generated using more extensive cultivation approaches, reveals a more complex microbial community than that obtained by conventional CF culture methods. To directly evaluate the cultivability of the airway microbiome, we examined six samples in depth using culture-enriched molecular profiling which combines culture-based methods with the molecular profiling methods of terminal restriction fragment length polymorphisms and 16S rRNA gene sequencing. We demonstrate that combining culture-dependent and culture-independent approaches enhances the sensitivity of either approach alone. Our techniques were able to cultivate 43 of the 48 families detected by deep sequencing; the five families recovered solely by culture-independent approaches were all present at very low abundance (<0.002% total reads. 46% of the molecular signatures detected by culture from the six patients were only identified in an anaerobic environment, suggesting that a large proportion of the cultured airway community is composed of obligate anaerobes. Most significantly, using 20 growth conditions per specimen, half of which included anaerobic cultivation and extended incubation times we demonstrate that the majority of bacteria present can be cultured.

  6. Anaerobic mineralization of toluene by enriched sediments with quinones and humus as terminal electron acceptors

    NARCIS (Netherlands)

    Cervantes, F.J.; Dijksma, W.; Duong-Dac, T.; Ivanova, A.; Lettinga, G.; Field, J.A.

    2001-01-01

    The anaerobic microbial oxidation of toluene to CO2 coupled to humus respiration was demonstrated by use of enriched anaerobic sediments from the Amsterdam petroleum harbor (APH) and the Rhine River. Both highly purified soil humic acids (HPSHA) and the humic quinone moiety model compound anthraquin

  7. 一个硫酸盐还原细菌富集物对丁草胺的厌氧降解%Anaerobic degradation of butachlor by sulfate-reducing bacteria enrichment culture

    Institute of Scientific and Technical Information of China (English)

    叶央芳; 杜宇峰

    2000-01-01

    An enrichment culture of sulfate-reducing bacteria,capable of anaerabic degrading butachlor,was obtained.The degradation kinetics of butachlor by the enrichment culture was determined and the optimum concentration of butachlor,the optimum pH and temperature for degradation of butachlor were observed..%通过多次富集培养,得到一个能有效厌氧降解丁草胺的硫酸盐还原细菌(SRB)富集物,并对该富集物的生长动力学以及生长的最适丁草胺浓度、最适pH和最适温度作了探讨.

  8. Cultural Enrichment through Community Action.

    Science.gov (United States)

    Wilson, O. J.

    This project was conceived as a technique for helping to eliminate a cultural void in the areas of art, music, and theatre in the service area of Western Kentucky University. To implement this concept, demonstrations were conducted in art, music, theatre, and in library and lecture resources in 16 counties over a four-year period. The attendance…

  9. Subculturing of a polychlorinated biphenyl-dechlorinating anaerobic enrichment on solid media

    Energy Technology Data Exchange (ETDEWEB)

    May, H.D.; Boyle, A.W.; Price, W.A. II.; Blake, C.K. (Celgene Corp., Warren, NJ (United States))

    1992-12-01

    An anaerobic culture capable of dechlorinating polychlorinated biphenyls was subcultured under strict anaerobic conditions on solid media containing sterilized river sediment. The dechlorination activity was transferred as a bacterial colony on a solid medium three times. After two transfers on solid medium, the culture was no longer methanogenic but still dechlorinated a mixture of tri- and tetrachlorobiphenyls. This demonstrates that anaerobic bacteria are responsible for the polychlorinated biphenyl dechlorination and can be grown without polychlorinated biphenyl on solid media.

  10. Metabolic characteristics of an aerobe isolated from a methylotrophic methanogenic enrichment culture

    Indian Academy of Sciences (India)

    Stephen V Rapheal; K R Swaminathan; K Lalitha

    2003-03-01

    An anaerobic methylotrophic methanogenic enrichment culture, with sustained metabolic characteristics, including that of methanation for over a decade, was the choice of the present study on interspecies interactions. Growth and methanation by the enrichment were suppressed in the presence of antibiotics, and no methanogen grown on methanol could be isolated using stringent techniques. The present study confirmed syntrophic metabolic interactions in this enrichment with the isolation of a strain of Pseudomonas sp. The organism had characteristic metabolic versatility in metabolizing a variety of substrates including alcohols, aliphatic acids, amino acids, and sugars. Anaerobic growth was favoured with nitrate in the growth medium. Cells grown anaerobically with methanol, revealed maximal nitrate reductase activity. Constitutive oxidative activity of the membrane system emerged from the high-specific oxygen uptake and nitrate reductase activities of the aerobically and anerobically grown cells respectively. Cells grown anaerobically on various alcohols effectively oxidized methanol in the presence of flavins, cofactor FAD and the methanogenic cofactor F420, suggesting a constitutive alcohol oxidizing capacity. In cells grown anaerobically on methanol, the rate of methanol oxidation with F420 was three times that of FAD. Efficient utilization of alcohols in the presence of F420 is a novel feature of the present study. The results suggest that utilization of methanol by the mixed culture would involve metabolic interactions between the Pseudomonas sp. and the methanogen(s). Methylotrophic, methanogenic partnership involving an aerobe is a novel feature hitherto unreported among anaerobic syntrophic associations and is of ecological significance.

  11. Utilization of aminoaromatic acids by a methanogenic enrichment culture and by a novel Citrobacter freundii strain

    NARCIS (Netherlands)

    Savelieva, O.; Kotova, I.; Roelofsen, W.; Stams, A.J.M.; Netrusov, A.

    2004-01-01

    Following incubation of mesophilic methanogenic floccular sludge from a lab-scale upflow anaerobic sludge bed reactor used to treat cattle manure wastewater, a stable 5-aminosalicylate-degrading enrichment culture was obtained. Subsequently, a Citrobacter freundii strain, WA1, was isolated from the

  12. 21 CFR 866.2330 - Enriched culture medium.

    Science.gov (United States)

    2010-04-01

    ... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Enriched culture medium. 866.2330 Section 866.2330...) MEDICAL DEVICES IMMUNOLOGY AND MICROBIOLOGY DEVICES Microbiology Devices § 866.2330 Enriched culture medium. (a) Identification. An enriched culture medium is a device that consists primarily of liquid...

  13. Effect of chlorate, molybdate, and shikimic acid on Salmonella enterica serovar Typhimurium in aerobic and anaerobic cultures.

    Science.gov (United States)

    Oliver, Christy E; Beier, Ross C; Hume, Michael E; Horrocks, Shane M; Casey, Thomas A; Caton, Joel S; Nisbet, David J; Smith, David J; Krueger, Nathan A; Anderson, Robin C

    2010-04-01

    Experiments were conducted to determine factors that affect sensitivity of Salmonella enterica serovar Typhimurium to sodium chlorate (5mM). In our first experiment, cultures grown without chlorate grew more rapidly than those with chlorate. An extended lag before logarithmic growth was observed in anaerobic but not aerobic cultures containing chlorate. Chlorate inhibition of growth during aerobic culture began later than that observed in anaerobic cultures but persisted once inhibition was apparent. Conversely, anaerobic cultures appeared to adapt to chlorate after approximately 10h of incubation, exhibiting rapid compensatory growth. In anaerobic chlorate-containing cultures, 20% of total viable counts were resistant to chlorate by 6h and had propagated to 100% resistance (>10(9)CFU mL(-1)) by 24h. In the aerobic chlorate-containing cultures, 12.9% of colonies had detectable resistance to chlorate by 6h, but only 1% retained detectable resistance at 24h, likely because these cultures had opportunity to respire on oxygen and were thus not enriched via the selective pressure of chlorate. In another study, treatment with shikimic acid (0.34 mM), molybdate (1mM) or their combination had little effect on aerobic or anaerobic growth of Salmonella in the absence of added chlorate. As observed in our earlier study, chlorate resistance was not detected in any cultures without added chlorate. Chlorate resistant Salmonella were recovered at equivalent numbers regardless of treatment after 8h of aerobic or anaerobic culture with added chlorate; however, by 24h incubation chlorate sensitivity was completely restored to aerobic but not anaerobic cultures treated with shikimic acid or molybdate but not their combination. Results indicate that anaerobic adaptation of S. Typhimurium to sodium chlorate during pure culture is likely due to the selective propagation of low numbers of cells exhibiting spontaneous resistance to chlorate and this resistance is not reversible by

  14. Characteristics of enriched cultures for bio-huff-`n`-puff tests at Jilin oil field

    Energy Technology Data Exchange (ETDEWEB)

    Xiu-Yuan Wang; Gang Dai; Yan-Fen Xue; Shu-Hua Xie [Institute of Microbiology, Beijing (China)] [and others

    1995-12-31

    Three enriched cultures (48, 15a, and 26a), selected from more than 80 soil and water samples, could grow anaerobically in the presence of crude oil at 30{degrees}C and could ferment molasses to gases and organic acids. Oil recovery by culture 48 in the laboratory model experiment was enhanced by 25.2% over the original reserves and by 53.7% over the residual reserves. Enriched culture 48 was composed of at least 4 species belonging to the genera Eubacterium, Fusobacterium, and Bacteroides. This enriched culture was used as inoculum for MEOR field trials at Jilin oil field with satisfactory results. The importance of the role of these isolates in EOR was confirmed by their presence and behavior in the fluids produced from the microbiologically treated reservoir.

  15. Enrichment and hydrogen production by marine anaerobic hydrogen-producing microflora

    Institute of Scientific and Technical Information of China (English)

    CAI JinLing; WANG GuangCe; LI YanChuan; ZHU DaLing; PAN GuangHua

    2009-01-01

    Acid,alkali,heat-shock,KNO3 and control pretreatment methods applied to anaerobic sludge were evaluated for their ability to selectively enrich the marine hydrogen-producing mixed microflora.Seawater culture medium was used as the substrate.The hydrogen yield of pretreated microflora was higher than that of the un-pretreated control (P<0.05).Among the pretreatment methods studied,heat-shock pretreatment yielded the greatest hydrogen production,which was 14.6 times that of the control.When the effect of initial pH on hydrogen production of heat-shock pretreated samples was studied,hydrogen was produced over the entire pH range (pH 4-10).The hydrogen yield peaked at initial pH 8 (79 mL/g sucrose) and then steadily decreased as the initial pH increased.Sucrose consumption was high at neutral initial pH.During the process of hydrogen production,pH decreased gradually,which indicated that the acquired microflora consisted of acidogenic bacteria.

  16. Microbial community in anaerobic hydrogen-producing microflora enriched from sludge compost.

    Science.gov (United States)

    Ueno, Y; Haruta, S; Ishii, M; Igarashi, Y

    2001-11-01

    Hydrogen production by thermophilic anaerobic microflora enriched from sludge compost was studied by using an artificial medium containing cellulose powder. Hydrogen gas was evolved with the formation of acetate, ethanol, and butyrate by decomposition of the cellulose powder. The hydrogen production yield was 2.0 mol/mol-hexose by either batch or chemostat cultivation. A medium that did not contain peptone demonstrated a lower hydrogen production yield of 1.0 mol/mol-hexose with less formation of butyrate. The microbial community in the microflora was investigated through isolation of the microorganisms by both plating and denaturing gradient gel electrophoresis (DGGE) of the' PCR-amplified V3 region of 16S rDNA. Sixty-eight microorganisms were isolated from the microflora and classified into nine distinct groups by genetic fingerprinting of the PCR-DGGE or by a random amplified polymorphic DNA analysis and determination of the partial sequence of 16S rDNA. Most of the isolates belonged to the cluster of the thermophilic Clostridium/Bacillus subphylum of low G+C gram-positive bacteria. Product formation by most of the isolated strains corresponded to that produced by the microflora. Thermoanaerobacterium thermosaccharolyticium was isolated in the enrichment culture with or without added peptone. and was detected with strong intensity by PCR-DGGE. Two other thermophilic cellulolytic microorganisms, Clostridium thermocellum and Clostridium cellulosi, were also detected by PCR-DGGE, although they could not be isolated. These findings imply that hydrogen production from cellulose by microflora is performed by a consortium of several species of microorganisms.

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

    KAUST Repository

    Meulepas, Roel J W

    2010-05-06

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

  18. Effect of inorganic carbon on anaerobic ammonium oxidation enriched in sequencing batch reactor

    Institute of Scientific and Technical Information of China (English)

    Liao Dexiang; Li Xiaoming; Yang Qi; Zeng Guangming; Guo Liang; Yue Xiu

    2008-01-01

    The present lab-scale research reveals the enrichment of anaerobic ammonium oxidation microorganism from methanogenic anaerobic granular sludge and the effect of inorganic carbon (sodium bicarbonate) on anaerobic ammonium oxidation. The enrichment of anammox bacteria was carried out in a 7.0-L SBR and the effect of bicarbonate on anammox was conducted in a 3.0-L SBR. Research results , especially the biomass, showed first signs of anammox activity after 54 d cultivation with synthetic wastewater, when the pH was controlled between 7.5 and 8.3, the temperature was 35℃. The anammox activity increased as the influent bicarbonate concentration increased from 1.0 to 1.5 g/L and then, was inhibited as the bicarbonate concentration approached 2.0 g/L. However, the activity could be restored by the reduction of bicarbonate concentration to 1.0 g/L, as shown by rapid conversion of ammonium, and nitrite and nitrate production with normal stoichiometry. The optimization of the bicarbonate concentration in the reactor could increase the anammox rate up to 66.4 mgN/(L·d).

  19. Enhancement of anaerobic methanogenesis at a short hydraulic retention time via bioelectrochemical enrichment of hydrogenotrophic methanogens.

    Science.gov (United States)

    Li, Yang; Zhang, Yaobin; Liu, Yiwen; Zhao, Zhiqiang; Zhao, Zisheng; Liu, Sitong; Zhao, Huimin; Quan, Xie

    2016-10-01

    Anaerobic digestion (AD) is an important energy strategy for converting organic waste to CH4. A major factor limiting the practical applicability of AD is the relatively long hydraulic retention time (HRT) which declines the treatment efficiency of digesters. A coupling process of anaerobic digestion and 'electromethanogenesis' was proposed to enhance anaerobic digestion at a short HRT in this study. Microorganisms analysis indicated that the electric-biological reactor enriched hydrogenotrophic methanogens in both cathodic biofilm and suspended sludge, helping achieve the high organic removal (71.0% vs 42.3% [control reactor]) and CH4 production (248.5mL/h vs 51.3mL/h), while the additional electric input was only accounted for 25.6% of the energy income from the increased CH4 production. This study demonstrated that a bioelectrochemical enhanced anaerobic reactor could improve the CH4 production and organic removal at a short HRT, providing an economically feasible scheme to treat wastewater.

  20. Nitrogen removal by autotrophic ammonium oxidizing bacteria enrichment under anaerobic conditions

    Directory of Open Access Journals (Sweden)

    Pongsak (Lek Noophan

    2008-07-01

    Full Text Available Sludge from an anoxic tank at the centralized wastewater treatment plant, Nong Khaem, Bangkok, Thailand, was inoculatedin an anaerobic sequencing batch reactor (ASBR. The optimal compositions and operating conditions of the stock of autotrophic ammonium oxidizing bacteria medium were determined. The process of oxidizing ammonium with bacteria under anaerobic conditions is often referred to as the Anammox process (NO2- to N2 gas, using NH4+ as the electron donor and NO2- as the electron acceptor. The startup period for the anammox culture took more than three months. With ammoniumand nitrite concentration ratios of 1:1.38 and 1:1.6, the nitrogen conversion rate zero order. Fluorescent in situ hybridization(FISH was used to identify specific autotrophic ammonium oxidizing bacteria (Nitrosomonas spp., Candidatus Brocadia anammoxidans, and Candidatus Kuenenia stuttgartiensis. Results from this work demonstrated a shift in the species of ammonium oxidizing bacteria from Nitrosomonas spp. to Candidati Brocadia anammoxidans and Kuenenia stuttgartiensis, with increased ammonium concentrations from 3 mM to 15 mM. Under NH4+:NO2- ratios of 1:1.38 and 1:1.6 the ammoniumoxidizing bacteria were able to remove both ammonium and nitrite simultaneously. The specific nitrogen removal rate of theanammox bacteria (Candidati Brocadia anammoxidans and Kuenenia stuttgartiensis was significantly higher than that of anaerobic ammonium oxidizing bacteria (Nitrosomonas spp.. Anaerobic ammonium oxidizing bacteria (Candidati Brocadia anammoxidans and Kuenenia stuttgartiensis are strict anaerobes.

  1. Biohydrogen production from desugared molasses (DM) using thermophilic mixed cultures immobilized on heat treated anaerobic sludge granules

    DEFF Research Database (Denmark)

    Kongjan, Prawit; O-Thong, Sompong; Angelidaki, Irini

    2011-01-01

    Hydrogen production from desugared molasses (DM) was investigated in both batch and continuous reactors using thermophilic mixed cultures enriched from digested manure by load shock (loading with DM concentration of 50.1 g-sugar/L) to suppress methanogens. H2 gas, free of methane, was produced...... by Thermoanaerobacterium spp., which are key players in fermentative hydrogen production of DM under thermophilic conditions. Furthermore, the granules in the UASB reactor were also significantly containing Thermoanaerobacterium spp. and phylum Firmecutes (most Clotridium, Bacillus and Desulfobacterium....... The enriched hydrogen producing mixed culture achieved from the 16.7 g-sugars/L DM batch cultivation was immobilized on heat treated anaerobic sludge granules in an up-flow anaerobic sludge blanket (UASB) reactor. The UASB reactor, operated at a hydraulic retention time (HRT) of 24 h fed with 16.7 g...

  2. Selective enrichment of Geobacter sulfurreducens from anaerobic granular sludge with quinones as terminal electron acceptors

    NARCIS (Netherlands)

    Cervantes-Carillo, F.J.; Duong Dac, T.; Ivanova, A.E.; Roest, de K.; Akkermans, A.D.L.; Lettinga, G.; Field, J.A.

    2003-01-01

    A quinone-respiring, enrichment culture derived from methanogenic granular sludge was phylogenetically characterized by using a combined cloning-denaturing gradient gel electrophoresis (DGGE) method, which revealed that the consortium developed was dominated by a single microorganism: 97% related, i

  3. Characterization of wheat straw-degrading anaerobic alkali-tolerant mixed cultures from soda lake sediments by molecular and cultivation techniques.

    Science.gov (United States)

    Porsch, Katharina; Wirth, Balázs; Tóth, Erika M; Schattenberg, Florian; Nikolausz, Marcell

    2015-09-01

    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.

  4. Hydrogen production from rice winery wastewater in an upflow anaerobic reactor by using mixed anaerobic cultures

    Energy Technology Data Exchange (ETDEWEB)

    Hanqing Yu; Zhenhu Zhu [University of Science and Technology, Hefei, Anhui (China). School of Chemistry and Materials; Wenrong Hu [Shandong Univ., Jinan (China). School of Resources and Environmental Engineering; Haisheng Zhang [Jingzi Wine Distillery Company, Shandong (China)

    2002-12-01

    Continuous production of hydrogen from the anaerobic acidogenesis of a high-strength rice winery wastewater by a mixed bacterial flora was demonstrated. The experiment was conducted in a 3.0-l upflow reactor to investigate individual effects of hydraulic retention time (HRT) (2-24 h), chemical oxygen demand (COD) concentration in wastewater (14-36 g COD/l), pH (4.5-6.0) and temperature (20-55{sup o}C) on bio-hydrogen production from the wastewater. The biogas produced under all test conditions was composed of mostly hydrogen (53-61%) and carbon dioxide (37-45%), but contained no detectable methane. Specific hydrogen production rate increased with wastewater concentration and temperature, but with a decrease in HRT. An optimum hydrogen production rate of 9.33 lH{sub 2}/gVSSd was achieved at an HRT of 2 h, COD of 34 g/l, pH 5.5 and 55{sup o}C. The hydrogen yield was in the range of 1.37-2.14 mol/mol-hexose. In addition to acetate, propionate and butyrate, ethanol was also present in the effluent as an aqueous product. The distribution of these compounds in the effluent was more sensitive to wastewater concentration, pH and temperature, but was less sensitive to HRT. This upflow reactor was shown to be a promising biosystem for hydrogen production from high-strength wastewaters by mixed anaerobic cultures. (Author)

  5. Methane enrichment digestion experiments at the anaerobic experimental test unit at Walt Disney World. Final report, March 1989-August 1990

    Energy Technology Data Exchange (ETDEWEB)

    Srivastava, V.J.; Hill, A.H.

    1993-06-01

    The goal of the project was to determine the technical feasibility of utilizing a novel concept in anaerobic digestion, in-situ methane enrichment digestion or MED for producing utility-grade gas from a pilot-scale anaerobic digester. MED tests conducted during this program consistently achieved digester product gas with a methane (CH4) content of greater than 90% (on a dry-, nitrogen-free basis). The MED concept, because it requires relatively simple equipment and modest energy input, has the potential to simplify gas cleanup requirements and substantially reduce the cost of converting wastes and biomass to pipeline quality gas.

  6. Anaerobic degradation of o-phenylphenol by mixed and pure cultures

    Energy Technology Data Exchange (ETDEWEB)

    Sembiring, T.; Winter, J.

    1989-07-01

    An anaerobic enrichment culture that degrade 0.4 mmol/l per day of o-phenylphenol was selected from sediment of a waste water pond of a sugar factory. From the consortium an o-phenylphenol-degrading bacterium, strain B10, was isolated. Strain B10 could not degrade other aromatic substances, including phenylacetic acid, benzoate, o-hydroxybenzoate, p-hydroxybenzoate and phenol. Best growth was observed with glucose, pyruvate, lactate, methanol and H/sub 2//CO/sub 2/ as substrates, o-Phenylphenol was slowly degraded if supplied as the only carbon source and was cometabolized in the presence of >5 mmol/l glucose. Strain B10 has not yet been assigned to a known species or family.

  7. Enhancement of fermentative hydrogen/ethanol production from cellulose using mixed anaerobic cultures

    Energy Technology Data Exchange (ETDEWEB)

    Lin, Chiu-Yue; Hung, Wen-Chin [BioHydrogen Laboratory, Department of Environmental Engineering and Science, Feng Chia University, Taichung 40724 (China)

    2008-07-15

    Batch tests were conducted to evaluate the enhancement of hydrogen/ethanol (EtOH) productivity using cow dung microflora to ferment {alpha}-cellulose and saccharification products (glucose and xylose). Hydrogen/ethanol production was evaluated based on hydrogen/ethanol yields (HY/EY) under 55 C at various initial pH conditions (5.5-9.0). Our test results indicate that cow dung sludge is a good mixed natural-microflora seed source for producing biohydrogen/ethanol from cellulose and xylose. The heat-pretreatment, commonly used to produce hydrogen more efficiently from hexose, applied to mixed anaerobic cultures did not help cow dung culture convert cellulose and xylose into hydrogen/ethanol. Instead of heat-pretreatment, the mixed culture received enrichments cultivated at 55 C for 4 days. Positive results were observed: hydrogen/ethanol production from fermenting cellulose and xylose was effectively enhanced at increases of 4.8 (ethanol) to 8 (hydrogen) and 2.4 (ethanol) to 15.6 (hydrogen) folds, respectively. In which, the ethanol concentration produced from xylose reached 4-4.4 g/L, an output comparable to that of using heat-treated sewage sludge and better than that (1.25-3 g/L) using pure cultures. Our test results show that for the enriched cultures the initial cultivation pH can affect hydrogen/ethanol production including HY, EY and liquid fermentation product concentration and distribution. These results were also concurred using a denaturing gradient gel electrophoresis analysis saying that both cultivation pH and substrate can affect the enriched cow dung culture microbial communities. The enriched cow dung culture had an optimal initial cultivation pH range of 7.6-8.0 with peak HY/EY values of 2.8 mmol-H{sub 2}/g-cellulose, 5.8 mmol-EtOH/g-cellulose, 0.3 mol-H{sub 2}/mol-xylose and 1 mol-EtOH/mol-xylose. However, a pH change of 0.5 units from the optimal values reduced hydrogen/ethanol production efficiency by 20%. Strategies based on the experimental

  8. The maximum specific hydrogen-producing activity of anaerobic mixed cultures: definition and determination

    Science.gov (United States)

    Mu, Yang; Yang, Hou-Yun; Wang, Ya-Zhou; He, Chuan-Shu; Zhao, Quan-Bao; Wang, Yi; Yu, Han-Qing

    2014-06-01

    Fermentative hydrogen production from wastes has many advantages compared to various chemical methods. Methodology for characterizing the hydrogen-producing activity of anaerobic mixed cultures is essential for monitoring reactor operation in fermentative hydrogen production, however there is lack of such kind of standardized methodologies. In the present study, a new index, i.e., the maximum specific hydrogen-producing activity (SHAm) of anaerobic mixed cultures, was proposed, and consequently a reliable and simple method, named SHAm test, was developed to determine it. Furthermore, the influences of various parameters on the SHAm value determination of anaerobic mixed cultures were evaluated. Additionally, this SHAm assay was tested for different types of substrates and bacterial inocula. Our results demonstrate that this novel SHAm assay was a rapid, accurate and simple methodology for determining the hydrogen-producing activity of anaerobic mixed cultures. Thus, application of this approach is beneficial to establishing a stable anaerobic hydrogen-producing system.

  9. Temperature effects on fermentative hydrogen production from xylose using mixed anaerobic cultures

    Energy Technology Data Exchange (ETDEWEB)

    Lin, Chiu-Yue; Wu, Chein-Chung [BioHydrogen Laboratory, Department of Water Resource Engineering, Feng Chia University, P.O. Box 25-123, Taichung 40724 (China); Hung, Chun-Hsiung [Department of Environmental Engineering, National Chung-Hsing University, 250 Kuo-Kuang Road, Taichung 40227 (China)

    2008-01-15

    Sewage sludge microflora were anaerobically cultivated in a chemostat-type anaerobic bioreactor at temperatures of 30-55{sup o}C, a pH of 7.1 and a hydraulic retention time of 12 h to determine the hydrogen production efficiency from xylose (20 g-COD/L). It was demonstrated that hydrogen production of the enriched sewage sludge microflora (dominated by Clostridia species) was temperature-dependent in hydrogen gas content, hydrogen yield, hydrogen production rate (HPR) and specific HPR with values of 25.1-42.2% (v/v), 0.4-1.4 mol-H{sub 2}/mol-xylose, 0.06-0.24 mol-H{sub 2}/L-day and 0.02-0.10 mol-H{sub 2}/g-VSS-day, respectively, and the above values peaked whaaen being operated at 50{sup o}C. A transition temperature of 45{sup o}C existed by having a lowest hydrogen production efficiency. Butyrate and ethanol were the major soluble metabolite products for thermophilic and mesophilic fermentation, respectively. The liquid metabolite concentration fractions and microbial community analysis indicate that the differences in hydrogen production efficiency between each tested temperature might relate to the shifts in metabolic pathway or microbial community. Thermodynamic analysis using HPR values and Arrhenius equation showed that the activation energy was 74.7 kJ/mol. Strategies based on temperature control for optimal hydrogen production from xylose using natural mixed cultures are proposed. (author)

  10. Simultaneous enrichment of denitrifying anaerobic methane-oxidizing microorganisms and anammox bacteria in a hollow-fiber membrane biofilm reactor.

    Science.gov (United States)

    Ding, Zhao-Wei; Lu, Yong-Ze; Fu, Liang; Ding, Jing; Zeng, Raymond J

    2017-01-01

    In this study, the coculture system of denitrifying anaerobic methane oxidation (DAMO) microbes and anaerobic ammonium oxidation (anammox) bacteria was successfully enriched in a hollow-fiber membrane biofilm reactor (HfMBR) using freshwater sediment as the inoculum. The maximal removal rates of nitrate and ammonium were 78 mg N/L/day (131 mg N/m(2)/day) and 26 mg N/L/day (43 mg N/m(2)/day), respectively. Due to the high rate of methane mass transfer in HfMBR, the activity of DAMO archaea continued to increase during the enrichment period, indicating that HfMBR could be a powerful tool to enrich DAMO microorganisms. Effects of partial methane pressure, temperature, and pH on the cocultures were obvious. However, the microbial activity in HfMBR could be recovered quickly after the shock change of environmental factors. Furthermore, the result also found that DAMO bacteria likely had a stronger competitive advantage than anammox bacteria under the operating conditions in this study. High-throughput sequencing 16S rRNA genes illustrated that the dominant microbes were NC10, Euryarchaeota, Proteobacteria, Planctomycetes, and Chlorobi with relative abundance of 38.8, 26.2, 13.78, 6.2, and 3.6 %, respectively.

  11. Batch fermentative hydrogen production by enriched mixed culture: Combination strategy and their microbial composition.

    Science.gov (United States)

    Sivagurunathan, Periyasamy; Sen, Biswarup; Lin, Chiu-Yue

    2014-02-01

    The effect of individual and combined mixed culture on dark fermentative hydrogen production performance was investigated. Mixed cultures from cow dung (C1), sewage sludge (C2), and pig slurry (C3) were enriched under strict anaerobic conditions at 37°C with glucose as the sole carbon source. Biochemical hydrogen production test in peptone-yeast-glucose (PYG) and basal medium was performed for individual mixed cultures (C1, C2 and C3) and their combinations (C1-C2, C2-C3, C1-C3 and C1-C2-C3) at a glucose concentration of 10 g/L, 37°C and initial pH 7. Maximum hydrogen yields (HY) of 2.0 and 1.86 [Formula: see text] by C2, and 1.98 and 1.95 mol(H2)/mol(glucose) by C2-C3 were obtained in PYG and basal medium, respectively. Butyrate and acetate were the major soluble metabolites produced by all the cultures, and the ratio of butyrate to acetate was ∼2 fold higher in basal medium than PYG medium, indicating strong influence of media formulation on glucose catabolism. The major hydrogen-producing bacterial strains, observed in all mixed cultures, belonged to Clostridium butyricum, C. saccharobutylicum, C. tertium and C. perfringens. The hydrogen production performance of the combined mixed culture (C2-C3) was further evaluated on beverage wastewater (10 g/L) at pH 7 and 37°C. The results showed an HY of 1.92 mol(H2)/mol(glucose-equivalent). Experimental evidence suggests that hydrogen fermentation by mixed culture combination could be a novel strategy to improve the HY from industrial wastewater.

  12. Dimethylamine biodegradation by mixed culture enriched from drinking water biofilter.

    Science.gov (United States)

    Liao, Xiaobin; Chen, Chao; Zhang, Jingxu; Dai, Yu; Zhang, Xiaojian; Xie, Shuguang

    2015-01-01

    Dimethylamine (DMA) is one of the important precursors of drinking water disinfection by-product N-nitrosodimethylamine (NDMA). Reduction of DMA to minimize the formation of carcinogenic NDMA in drinking water is of practical importance. Biodegradation plays a major role in elimination of DMA pollution in the environment, yet information on DMA removal by drinking water biofilter is still lacking. In this study, microcosms with different treatments were constructed to investigate the potential of DMA removal by a mixed culture enriched from a drinking water biofilter and the effects of carbon and nitrogen sources. DMA could be quickly mineralized by the enrichment culture. Amendment of a carbon source, instead of a nitrogen source, had a profound impact on DMA removal. A shift in bacterial community structure was observed with DMA biodegradation, affected by carbon and nitrogen sources. Proteobacteria was the predominant phylum group in DMA-degrading microcosms. Microorganisms from a variety of bacterial genera might be responsible for the rapid DMA mineralization.

  13. Time-to-positivity-based discrimination between Enterobacteriaceae, Pseudomonas aeruginosa and strictly anaerobic Gram-negative bacilli in aerobic and anaerobic blood culture vials.

    Science.gov (United States)

    Defrance, Gilles; Birgand, Gabriel; Ruppé, Etienne; Billard, Morgane; Ruimy, Raymond; Bonnal, Christine; Andremont, Antoine; Armand-Lefèvre, Laurence

    2013-05-01

    Time-to-positivity (TTP) of first positive blood cultures growing Gram-negative bacilli (GNB) was investigated. When anaerobic vials were positive first, TTP ≤ 18 h differentiated Enterobacteriaceae from strict anaerobic Gram-negative bacilli (PPV 98.8%). When the aerobic ones were first, TTP ≤ 13 h differentiated Enterobacteriaceae from Pseudomonas aeruginosa and other GNB (PPV 80.8%).

  14. An automated blood culture system: the detection of anaerobic bacteria using a Malthus Microbiological Growth Analyser.

    Science.gov (United States)

    McMaster, J P; Barr, J G; Campbell, R R; Bennett, R B; Smyth, E T

    1985-10-01

    The Malthus Microbiological Growth Analyser has proved to be sensitive in detecting conductivity changes due to anaerobic metabolism in a number of widely used blood culture media. Freshly prepared cooked meat media and Thiol medium yielded the greatest gross conductivity changes, and were more sensitive of anaerobic metabolism than other media. Failure of the instrument to detect anaerobic metabolism was a problem particularly associated with growth in the thioglycollate medium. False positive detections of growth were attributed to a number of factors including electrode instability (6.0%) and bacterial contamination (8.75%).

  15. Anaerobic co-culture of mesenchymal stem cells and anaerobic pathogens - a new in vitro model system.

    Directory of Open Access Journals (Sweden)

    Katja Kriebel

    Full Text Available BACKGROUND: Human mesenchymal stem cells (hMSCs are multipotent by nature and are originally isolated from bone marrow. In light of a future application of hMSCs in the oral cavity, a body compartment with varying oxygen partial pressures and an omnipresence of different bacterial species i.e. periodontitis pathogens, we performed this study to gain information about the behavior of hMSC in an anaerobic system and the response in interaction with oral bacterial pathogens. METHODOLOGY/PRINCIPAL FINDINGS: We established a model system with oral pathogenic bacterial species and eukaryotic cells cultured in anaerobic conditions. The facultative anaerobe bacteria Fusobacterium nucleatum, Porphyromonas gingivalis and Aggregatibacter actinomycetemcomitans were studied. Their effects on hMSCs and primary as well as permanent gingival epithelial cells (Ca9-22, HGPEC were comparatively analyzed. We show that hMSCs cope with anoxic conditions, since 40% vital cells remain after 72 h of anaerobic culture. The Ca9-22 and HGPEC cells are significantly more sensitive to lack of oxygen. All bacterial species reveal a comparatively low adherence to and internalization into hMSCs (0.2% and 0.01% of the initial inoculum, respectively. In comparison, the Ca9-22 and HGPEC cells present better targets for bacterial adherence and internalization. The production of the pro-inflammatory chemokine IL-8 is higher in both gingival epithelial cell lines compared to hMSCs and Fusobacterium nucleatum induce a time-dependent cytokine secretion in both cell lines. Porphyromonas gingivalis is less effective in stimulating secretion of IL-8 in the co-cultivation experiments. CONCLUSIONS/SIGNIFICANCE: HMSCs are suitable for use in anoxic regions of the oral cavity. The interaction with local pathogenic bacteria does not result in massive pro-inflammatory cytokine responses. The test system established in this study allowed further investigation of parameters prior to set up of

  16. Diversity of reductive dehalogenase genes from environmental samples and enrichment cultures identified with degenerate primer PCR screens.

    Directory of Open Access Journals (Sweden)

    Laura Audrey Hug

    2013-11-01

    Full Text Available Reductive dehalogenases are the critical enzymes for anaerobic organohalide respiration, a microbial metabolic process that has been harnessed for bioremediation efforts to resolve chlorinated solvent contamination in groundwater and is implicated in the global halogen cycle. Reductive dehalogenase sequence diversity is informative for the dechlorination potential of the site or enrichment culture. A suite of degenerate PCR primers targeting a comprehensive curated set of reductive dehalogenase genes was designed and applied to twelve DNA samples extracted from contaminated and pristine sites, as well as six enrichment cultures capable of reducing chlorinated compounds to non-toxic end-products. The amplified gene products from four environmental sites and two enrichment cultures were sequenced using Illumina HiSeq, and the reductive dehalogenase complement of each sample determined. The results indicate that the diversity of the reductive dehalogenase gene family is much deeper than is currently accounted for: one-third of the translated proteins have less than 70% pairwise amino acid identity to database sequences. Approximately 60% of the sequenced reductive dehalogenase genes were broadly distributed, being identified in four or more samples, and often in previously sequenced genomes as well. In contrast, 17% of the sequenced reductive dehalogenases were unique, present in only a single sample and bearing less than 90% pairwise amino acid identity to any previously identified proteins. Many of the broadly distributed reductive dehalogenases are uncharacterized in terms of their substrate specificity, making these intriguing targets for further biochemical experimentation. Finally, comparison of samples from a contaminated site and an enrichment culture derived from the same site eight years prior allowed examination of the effect of the enrichment process.

  17. Hydrogenase activity in aged, nonviable Desulfovibrio vulgaris cultures and its significance in anaerobic biocorrosion.

    Science.gov (United States)

    Chatelus, C; Carrier, P; Saignes, P; Libert, M F; Berlier, Y; Lespinat, P A; Fauque, G; Legall, J

    1987-07-01

    Batch cultures of Desulfovibrio vulgaris stored at 32 degrees C for 10 months have been found to retain 50% of the hydrogenase activity of a 1-day culture. The hydrogenase found in old cultures needs reducing conditions for its activation. Viable cell counts are negative after 6 months, showing that the hydrogenase activity does not depend on the presence of viable cells. These observations are of importance in the understanding of anaerobic biocorrosion of metals caused by depolarization phenomena.

  18. Selective enrichment media bias the types of Salmonella enterica strains isolated from mixed strain cultures and complex enrichment broths.

    Science.gov (United States)

    Gorski, Lisa

    2012-01-01

    For foodborne outbreak investigations it can be difficult to isolate the relevant strain from food and/or environmental sources. If the sample is contaminated by more than one strain of the pathogen the relevant strain might be missed. In this study mixed cultures of Salmonella enterica were grown in one set of standard enrichment media to see if culture bias patterns emerged. Nineteen strains representing four serogroups and ten serotypes were compared in four-strain mixtures in Salmonella-only and in cattle fecal culture enrichment backgrounds using Salmonella enrichment media. One or more strain(s) emerged as dominant in each mixture. No serotype was most fit, but strains of serogroups C2 and E were more likely to dominate enrichment culture mixtures than strains of serogroups B or C1. Different versions of Rappaport-Vassiliadis (RV) medium gave different patterns of strain dominance in both Salmonella-only and fecal enrichment culture backgrounds. The fittest strains belonged to serogroups C1, C2, and E, and included strains of S. Infantis, S. Thompson S. Newport, S. 6,8:d:-, and S. Give. Strains of serogroup B, which included serotypes often seen in outbreaks such as S. Typhimurium, S. Saintpaul, and S. Schwarzengrund were less likely to emerge as dominant strains in the mixtures when using standard RV as part of the enrichment. Using a more nutrient-rich version of RV as part of the protocol led to a different pattern of strains emerging, however some were still present in very low numbers in the resulting population. These results indicate that outbreak investigations of food and/or other environmental samples should include multiple enrichment protocols to ensure isolation of target strains of Salmonella.

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

    Science.gov (United States)

    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).

  20. Carbon capture and biogas enhancement by carbon dioxide enrichment of anaerobic digesters treating sewage sludge or food waste.

    Science.gov (United States)

    Bajón Fernández, Y; Soares, A; Villa, R; Vale, P; Cartmell, E

    2014-05-01

    The increasing concentration of carbon dioxide (CO2) in the atmosphere and the stringent greenhouse gases (GHG) reduction targets, require the development of CO2 sequestration technologies applicable for the waste and wastewater sector. This study addressed the reduction of CO2 emissions and enhancement of biogas production associated with CO2 enrichment of anaerobic digesters (ADs). The benefits of CO2 enrichment were examined by injecting CO2 at 0, 0.3, 0.6 and 0.9 M fractions into batch ADs treating food waste or sewage sludge. Daily specific methane (CH4) production increased 11-16% for food waste and 96-138% for sewage sludge over the first 24h. Potential CO2 reductions of 8-34% for sewage sludge and 3-11% for food waste were estimated. The capacity of ADs to utilise additional CO2 was demonstrated, which could provide a potential solution for onsite sequestration of CO2 streams while enhancing renewable energy production.

  1. Biological hydrogen production from probiotic wastewater as substrate by selectively enriched anaerobic mixed microflora

    Energy Technology Data Exchange (ETDEWEB)

    Sivaramakrishna, D.; Sreekanth, D.; Himabindu, V. [Centre for Environment, Institute of Science and Technology, Jawaharlal Nehru Technological University, Kukatpally, Hyderabad 500072, Andhra Pradesh (India); Anjaneyulu, Y. [TLGVRC, JSU Box 18739, JSU, Jackson, MS 32917-0939 (United States)

    2009-03-15

    Biohydrogen production from probiotic wastewater using mixed anaerobic consortia is reported in this paper. Batch tests are carried out in a 5.0 L batch reactor under constant mesophillic temperature (37 C). The maximum hydrogen yield 1.8 mol-hydrogen/mol-carbohydrate is obtained at an optimum pH of 5.5 and substrate concentration 5 g/L. The maximum hydrogen production rate is 168 ml/h. The hydrogen content in the biogas is more than 65% and no significant methane is observed throughout the study. In addition to hydrogen, acetate, propionate, butyrate and ethanol are found to be the main by-products in the metabolism of hydrogen fermentation. (author)

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

    DEFF Research Database (Denmark)

    Wang, Han; Zhang, Yifeng; Angelidaki, Irini

    2016-01-01

    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......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...... more tolerant to high ammonia levels (≥5 g NH4+-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....

  3. Culturing aerobic and anaerobic bacteria and mammalian cells with a microfluidic differential oxygenator.

    Science.gov (United States)

    Lam, Raymond H W; Kim, Min-Cheol; Thorsen, Todd

    2009-07-15

    In this manuscript, we report on the culture of anaerobic and aerobic species within a disposable multilayer polydimethylsiloxane (PDMS) microfluidic device with an integrated differential oxygenator. A gas-filled microchannel network functioning as an oxygen-nitrogen mixer generates differential oxygen concentration. By controlling the relative flow rate of the oxygen and nitrogen input gases, the dissolved oxygen (DO) concentration in proximal microchannels filled with culture media are precisely regulated by molecular diffusion. Sensors consisting of an oxygen-sensitive dye embedded in the fluid channels permit dynamic fluorescence-based monitoring of the DO concentration using low-cost light-emitting diodes. To demonstrate the general utility of the platform for both aerobic and anaerobic culture, three bacteria with differential oxygen requirements (E. coli, A. viscosus, and F. nucleatum), as well as a model mammalian cell line (murine embryonic fibroblast cells (3T3)), were cultured. Growth characteristics of the selected species were analyzed as a function of eight discrete DO concentrations, ranging from 0 ppm (anaerobic) to 42 ppm (fully saturated).

  4. Composition of the bacterial community degrading Phaeocystis mucopolysaccharides in enrichment cultures

    NARCIS (Netherlands)

    Janse, Ingmar; Zwart, Gabriel; Maarel, Marc J.E.C. van der; Gottschal, Jan C.

    2000-01-01

    As described recently, mucopolysaccharides of the marine microalga Phaeocystis can be degraded in enrichment cultures. In this paper we report on the characterization of the microbial community in such enrichments. Denaturing gradient gel electrophoresis (DGGE) profiles that were obtained during muc

  5. Atrazine and its metabolites degradation in mineral salts medium and soil using an enrichment culture.

    Science.gov (United States)

    Kumar, Anup; Singh, Neera

    2016-03-01

    An atrazine-degrading enrichment culture was used to study degradation of atrazine metabolites viz. hydroxyatrazine, deethylatrazine, and deisopropylatrazine in mineral salts medium. Results suggested that the enrichment culture was able to degrade only hydroxyatrazine, and it was used as the sole source of carbon and nitrogen. Hydroxyatrazine degradation slowed down when sucrose and/or ammonium hydrogen phosphate were supplemented as the additional sources of carbon and nitrogen, respectively. The enrichment culture could degrade high concentrations of atrazine (up to 110 μg/mL) in mineral salts medium, and neutral pH was optimum for atrazine degradation. Further, except in an acidic soil, enrichment culture was able to degrade atrazine in three soil types having different physico-chemical properties. Raising the pH of acidic soil to neutral or alkaline enabled the enrichment culture to degrade atrazine suggesting that acidic pH inhibited atrazine-degrading ability. The study suggested that the enrichment culture can be successfully utilized to achieve complete degradation of atrazine and its persistent metabolite hydroxyatrazine in the contaminated soil and water.

  6. Enrichment methodology to increase the positivity of cultures from body fluids

    Directory of Open Access Journals (Sweden)

    Alessandra Valle Daur

    2006-12-01

    Full Text Available Isolation and identification of etiological agents found in body fluids can be of critical importance for the recovery of patients suffering from potentially-severe infections, which are often followed by serious sequels. Eighty-two samples of different body fluids were analyzed using two different methods: (1 the conventional culture method (agar plating and (2 the enrichment culture technique, using the Bact/Alert® blood culture bottle. The number of positive cultures increased on average from 9.7% to 23.1% with the enrichment culture technique. Pseudomonas aeruginosa, Escherichia coli and Staphylococcus aureus were the most frequently isolated bacteria. The enrichment method could provide a more accurate means the identifying etiological agents.

  7. Co-occurrence of Methanosarcina mazei and Geobacteraceae in an iron(III-reducing enrichment culture

    Directory of Open Access Journals (Sweden)

    Shiling eZheng

    2015-09-01

    Full Text Available Methanosaeta harundinacea and Methanosarcina barkeri, known as classic acetoclastic methanogens, are capable of directly accepting electrons from Geobacter metallireducens for the reduction of carbon dioxide to methane, having been revealed as direct interspecies electron transfer (DIET in the laboratory co-cultures. However, whether their co-occurrences are ubiquitous in the iron (III-reducing environments and the other species of acetoclastic methanogens such as Methanosarcina mazei are capable of DIET are still unknown. Instead of initiating the co-cultures with pure cultures, two-step cultivation was employed to selectively enrich iron (III-reducing microorganisms in a coastal gold mining river, Jiehe River, with rich iron content in the sediments. First, iron (III reducers including Geobacteraceae were successfully enriched by 3-months successive culture on amorphous Fe(III oxides as electron acceptor and acetate as electron donor. High-throughput Illumina sequencing, terminal restriction fragment length polymorphism (T-RFLP and clone library analysis based on 16S rRNA genes revealed that the enrichment cultures actively contained the bacteria belong to Geobacteraceae and Bacilli, exclusively dominated by the archaea belong to Methanosarcinaceae. Second, the enrichment cultures including methanogens and Geobacteraceae were transferred with ethanol as alternative electron donor. Remarkably, aggregates were successively formed in the enrichments after three transfers. The results revealed by RNA-based analysis demonstrate that the co-occurrence of Methanosarcina mazei and Geobacteraceae in an iron (III-reducing enrichment culture. Furthermore, the aggregates, as close physical contact, formed in the enrichment culture, indicate that DIET could be a possible option for interspecies electron transfer in the aggregates.

  8. Production of functional killer protein in batch cultures upon a shift from aerobic to anaerobic conditions

    Directory of Open Access Journals (Sweden)

    Gildo Almeida da Silva

    2011-06-01

    Full Text Available The aim of this work was to study the production of functional protein in yeast culture. The cells of Saccharomyces cerevisiae Embrapa 1B (K+R+ killed a strain of Saccharomyces cerevisiae Embrapa 26B (K-R-in grape must and YEPD media. The lethal effect of toxin-containing supernatant and the effect of aeration upon functional killer production and the correlation between the products of anaerobic metabolism and the functional toxin formation were evaluated. The results showed that at low sugar concentration, the toxin of the killer strain of Sacch. cerevisiae was only produced under anaerobic conditions . The system of killer protein production showed to be regulated by Pasteur and Crabtree effects. As soon as the ethanol was formed, the functional killer toxin was produced. The synthesis of the active killer toxin seemed to be somewhat associated with the switch to fermentation process and with concomitant alcohol dehydrogenase (ADH activity.

  9. Isolation, Characterization and Identification of Thiram-degrading Microorganisms from Soil Enrichment Cultures

    OpenAIRE

    ŞAHİN, Nurettin

    2000-01-01

    Mixed microbial cultures were obtained by enrichment from soil samples collected from the Gediz basin. A number of species of bacteria and fungi were isolated and partially characterized from enrichment clutures containing the fungicide thiram. According to their responses in morphological and biochemical tests fungi were identified as Aspergillus niger, A. flavus and Penicilium steckii. Bacterial isolates were assigned to the genera: Bacillus, Arthrobacter, Moraxella-like Acinetobacter a...

  10. Differential gene expression profiling of enriched human spermatogonia after short- and long-term culture.

    Science.gov (United States)

    Conrad, Sabine; Azizi, Hossein; Hatami, Maryam; Kubista, Mikael; Bonin, Michael; Hennenlotter, Jörg; Renninger, Markus; Skutella, Thomas

    2014-01-01

    This study aimed to provide a molecular signature for enriched adult human stem/progenitor spermatogonia during short-term (differentiation/spermatogenesis pathway were highly expressed in enriched short-term cultured spermatogonia. After long-term culture, a proportion of cells retained and aggravated the "spermatogonial" gene expression profile with the expression of germ and pluripotency-associated genes, while in the majority of long-term cultured cells this molecular profile, typical for the differentiation pathway, was reduced and more genes related to the extracellular matrix production and attachment were expressed. The approach we provide here to study the molecular status of in vitro cultured spermatogonia may be important to optimize the culture conditions and to evaluate the germ cell plasticity in the future.

  11. Detection of Salmonella invA gene in shrimp enrichment culture by polymerase chain reaction.

    Science.gov (United States)

    Upadhyay, Bishnu Prasad; Utrarachkij, Fuangfa; Thongshoob, Jarinee; Mahakunkijcharoen, Yuvadee; Wongchinda, Niracha; Suthienkul, Orasa; Khusmith, Srisin

    2010-03-01

    Contamination of seafood with salmonellae is a major public health concern. Detection of Salmonella by standard culture methods is time consuming. In this study, an enrichment culture step prior to polymerase chain reaction (PCR) was applied to detect 284 bp fragment of Salmonella invA in comparison with the conventional culture method in 100 shrimp samples collected from four different shrimp farms and fresh food markets around Bangkok. Samples were pre-enriched in non-selective lactose broth (LB) and selective tetrathionate broth (TTB). PCR detection limit was 10 pg and 10(4) cfu/ml of viable salmonellae with 100% specificity. PCR assay detected 19 different Salmonella serovars belonging to 8 serogroups (B, C1, C2-C3, D1, E1, E4 and K) commonly found in clinical and environmental samples in Thailand. The detection rate of PCR following TTB enrichment (24%) was higher than conventional culture method (19%). PCR following TTB, but not in LB enrichment allowed salmonella detection with 84% sensitivity, 90% specificity and 89% accuracy. Shrimp samples collected from fresh food markets had higher levels of contaminated salmonellae than those from shrimp farms. The results indicated that incorporation of an enrichment step prior to PCR has the potential to be applied for detection of naturally contaminated salmonellae in food, environment and clinical samples.

  12. Differential Gene Expression Profiling of Enriched Human Spermatogonia after Short- and Long-Term Culture

    Directory of Open Access Journals (Sweden)

    Sabine Conrad

    2014-01-01

    Full Text Available This study aimed to provide a molecular signature for enriched adult human stem/progenitor spermatogonia during short-term (<2 weeks and long-term culture (up to more than 14 months in comparison to human testicular fibroblasts and human embryonic stem cells. Human spermatogonia were isolated by CD49f magnetic activated cell sorting and collagen−/laminin+ matrix binding from primary testis cultures obtained from ten adult men. For transcriptomic analysis, single spermatogonia-like cells were collected based on their morphology and dimensions using a micromanipulation system from the enriched germ cell cultures. Immunocytochemical, RT-PCR and microarray analyses revealed that the analyzed populations of cells were distinct at the molecular level. The germ- and pluripotency-associated genes and genes of differentiation/spermatogenesis pathway were highly expressed in enriched short-term cultured spermatogonia. After long-term culture, a proportion of cells retained and aggravated the “spermatogonial” gene expression profile with the expression of germ and pluripotency-associated genes, while in the majority of long-term cultured cells this molecular profile, typical for the differentiation pathway, was reduced and more genes related to the extracellular matrix production and attachment were expressed. The approach we provide here to study the molecular status of in vitro cultured spermatogonia may be important to optimize the culture conditions and to evaluate the germ cell plasticity in the future.

  13. Breast Cancer Stem Cell Culture and Enrichment Using Poly(ε-Caprolactone Scaffolds

    Directory of Open Access Journals (Sweden)

    Sònia Palomeras

    2016-04-01

    Full Text Available The cancer stem cell (CSC population displays self-renewal capabilities, resistance to conventional therapies, and a tendency to post-treatment recurrence. Increasing knowledge about CSCs’ phenotype and functions is needed to investigate new therapeutic strategies against the CSC population. Here, poly(ε-caprolactone (PCL, a biocompatible polymer free of toxic dye, has been used to fabricate scaffolds, solid structures suitable for 3D cancer cell culture. It has been reported that scaffold cell culture enhances the CSCs population. A RepRap BCN3D+ printer and 3 mm PCL wire were used to fabricate circular scaffolds. PCL design and fabrication parameters were first determined and then optimized considering several measurable variables of the resulting scaffolds. MCF7 breast carcinoma cell line was used to assess scaffolds adequacy for 3D cell culture. To evaluate CSC enrichment, the Mammosphere Forming Index (MFI was performed in 2D and 3D MCF7 cultures. Results showed that the 60° scaffolds were more suitable for 3D culture than the 45° and 90° ones. Moreover, 3D culture experiments, in adherent and non-adherent conditions, showed a significant increase in MFI compared to 2D cultures (control. Thus, 3D cell culture with PCL scaffolds could be useful to improve cancer cell culture and enrich the CSCs population.

  14. Using Enrichment Clusters to Address the Needs of Culturally and Linguistically Diverse Learners

    Science.gov (United States)

    Allen, Jennifer K.; Robbins, Margaret A.; Payne, Yolanda Denise; Brown, Katherine Backes

    2016-01-01

    Using data from teacher interviews, classroom observations, and a professional development workshop, this article explains how one component of the schoolwide enrichment model (SEM) has been implemented at a culturally diverse elementary school serving primarily Latina/o and African American students. Based on a broadened conception of giftedness,…

  15. CULTURAL ENRICHMENT THROUGH COMMUNITY ACTION PROJECT. ANNUAL REPORT FOR PERIOD ENDING JULY 1, 1967. (TITLE SUPPLIED).

    Science.gov (United States)

    WILSON, O.J.

    WESTERN KENTUCKY UNIVERSITY INITIATED A PROJECT TO DETERMINE THE CULTURAL NEEDS AND INTERESTS OF THE AREA IT SERVES. IN PHASE ONE, CITY AND COUNTY REPRESENTATIVES ATTENDED A SYMPOSIUM ON ART, MUSIC, LIBRARY, LECTURE, AND THEATER RESOURCES AND FORMED A REGIONAL ADVISORY COUNCIL FOR COMMUNITY ENRICHMENT. PHASE TWO WAS A WORKSHOP TO HELP COUNCIL…

  16. 血培养厌氧菌实验室鉴定%ANAEROBIC BLOOD CULTURE OF LABORATORY IDENTIFICATION

    Institute of Scientific and Technical Information of China (English)

    郭素芳; 王俊瑞; 范文斌; 福泉; 张军力

    2015-01-01

    目的::通过血培养厌氧菌病例的临床及实验室检测资料,掌握厌氧菌鉴定方法,探讨实验室开展厌氧菌检测的重要性。方法:观察血培养仪厌氧血培养瓶阳性报警曲线,转种厌氧血琼脂厌氧环境培养,观察茵落形态,涂片革兰氏染色,VITIE 2鉴定到种。结果:12份血培养出厌氧菌13株。其中2份为需氧菌和厌氧菌混合感染,分别是脆弱拟杆菌混合血液链球菌和中间链球菌;1份血培养分离出2种厌氧菌分别为脆弱拟杆菌和梭形梭菌;其他血培养分别培养出产气荚膜梭菌、脆弱拟杆菌、单形拟杆菌等。结论:厌氧菌感染多为混合感染,且培养、鉴定要求条件较高。加强对厌氧菌感染的认识和实验室检测,对临床诊断、治疗以及合理使用抗生素具有指导作用。%Objective:By the case of anaerobic blood culture of clinical and laboratory test data,to master anaerobes identification methods and explore the importance of laboratory testing of anaerobic bacteria. Methods:Observe positive anaerobic blood culture bottles alarm curve in blood culture system,turn kind of anaerobic blood agar and anaerobic environment to culture,observing colony mor-phology,smearing by Gram staln and using VITIE 2 identified to species. Results:There are 13 anaerobic bacteria stralns of 12 cases of blood culture. Including 2 cases of aerobic and anaerobic mixed infections,respectively are bacteroides fragilis mixed with Streptococcus sanguis and Streptococcus in-termadius. One case of blood culture isolate two kinds of anaerobic bacteria were Bacteroides fragilis and Clostridium fusiform. Other blood cultures were isolated clostridium perfringens, bacteroides fragilis,Bacteroides uniformis and so on. Conclusion:We conclude that anaerobic bacteria infections are mostly mixed infection, the culture and identification of anaerobic bacteria requires a higher condition. Enhance understanding of anaerobic infections

  17. Chemical characterization and anaerobic biodegradability of hydrothermal liquefaction aqueous products from mixed-culture wastewater algae.

    Science.gov (United States)

    Tommaso, Giovana; Chen, Wan-Ting; Li, Peng; Schideman, Lance; Zhang, Yuanhui

    2015-02-01

    This study examined the chemical characteristics and the anaerobic degradability of the aqueous product from hydrothermal liquefaction (HTL-ap) from the conversion of mixed-culture algal biomass grown in a wastewater treatment system. The effects of the HTL reaction times from 0 to 1.5 h, and reaction temperatures from 260 °C to 320 °C on the anaerobic degradability of the HTL-ap were quantified using biomethane potential assays. Comparing chemical oxygen demand data for HTL-ap from different operating conditions, indicated that organic matter may partition from organic phase to aqueous phase at 320 °C. Moderate lag phase and the highest cumulative methane production were observed when HTL-ap was obtained at 320 °C. The longest lag phase and the smallest production rate were observed in the process fed with HTL-ap obtained at 300 °C. Nevertheless, after overcoming adaptation issues, this HTL-ap led to the second highest accumulated specific methane production. Acetogenesis was identified as a possible rate-limiting pathway.

  18. Quantitative proteome and transcriptome analysis of the archaeon Thermoplasma acidophilum cultured under aerobic and anaerobic conditions.

    Science.gov (United States)

    Sun, Na; Pan, Cuiping; Nickell, Stephan; Mann, Matthias; Baumeister, Wolfgang; Nagy, István

    2010-09-03

    A comparative proteome and transcriptome analysis of Thermoplasma acidophilum cultured under aerobic and anaerobic conditions has been performed. One-thousand twenty-five proteins were identified covering 88% of the cytosolic proteome. Using a label-free quantitation method, we found that approximately one-quarter of the identified proteome (263 proteins) were significantly induced (>2 fold) under anaerobic conditions. Thirty-nine macromolecular complexes were identified, of which 28 were quantified and 15 were regulated under anaerobiosis. In parallel, a whole genome cDNA microarray analysis was performed showing that the expression levels of 445 genes were influenced by the absence of oxygen. Interestingly, more than 40% of the membrane protein-encoding genes (145 out of 335 ORFs) were up- or down-regulated at the mRNA level. Many of these proteins are functionally associated with extracellular protein or peptide degradation or ion and amino acid transport. Comparison of the transcriptome and proteome showed only a weak positive correlation between mRNA and protein expression changes, which is indicative of extensive post-transcriptional regulatory mechanisms in T. acidophilum. Integration of transcriptomics and proteomics data generated hypotheses for physiological adaptations of the cells to anaerobiosis, and the quantitative proteomics data together with quantitative analysis of protein complexes provide a platform for correlation of MS-based proteomics studies with cryo-electron tomography-based visual proteomics approaches.

  19. Enrichment of cancer stem cell-like cells by culture in alginate gel beads.

    Science.gov (United States)

    Xu, Xiao-xi; Liu, Chang; Liu, Yang; Yang, Li; Li, Nan; Guo, Xin; Sun, Guang-wei; Ma, Xiao-jun

    2014-05-10

    Cancer stem cells (CSCs) are most likely the reason of cancer reoccurrence and metastasis. For further elucidation of the mechanism underlying the characteristics of CSCs, it is necessary to develop efficient culture systems to culture and expand CSCs. In this study, a three-dimensional (3D) culture system based on alginate gel (ALG) beads was reported to enrich CSCs. Two cell lines derived from different histologic origins were encapsulated in ALG beads respectively and the expansion of CSCs was investigated. Compared with two-dimensional (2D) culture, the proportion of cells with CSC-like phenotypes was significantly increased in ALG beads. Expression levels of CSC-related genes were greater in ALG beads than in 2D culture. The increase of CSC proportion after being cultured within ALG beads was further confirmed by enhanced tumorigenicity in vivo. Moreover, increased metastasis ability and higher anti-cancer drug resistance were also observed in 3D-cultured cells. Furthermore, we found that it was hypoxia, through the upregulation of hypoxia-inducible factors (HIFs) that occurred in ALG beads to induce the increasing of CSC proportion. Therefore, ALG bead was an efficient culture system for CSC enrichment, which might provide a useful platform for CSC research and promote the development of new anti-cancer therapies targeting CSCs.

  20. Treatment and electricity harvesting from sulfate/sulfide-containing wastewaters using microbial fuel cell with enriched sulfate-reducing mixed culture

    Energy Technology Data Exchange (ETDEWEB)

    Lee, Duu-Jong, E-mail: cedean@mail.ntust.edu.tw [Department of Chemical Engineering, National Taiwan University, Taipei, Taiwan (China); Department of Chemical Engineering, National Taiwan University of Science and Technology, Taipei, Taiwan (China); Lee, Chin-Yu [Department of Chemical Engineering, National Taiwan University, Taipei, Taiwan (China); Chang, Jo-Shu [Department of Chemical Engineering, National Cheng Kung University, Tainan, Taiwan (China); Center for Bioscience and Biotechnology, National Cheng Kung University, Tainan, Taiwan (China); Research Center for Energy Technology and Strategy, National Cheng Kung University, Tainan, Taiwan (China)

    2012-12-15

    Highlights: Black-Right-Pointing-Pointer We started up microbial fuel cell (MFC) using enriched sulfate-reducing mixed culture. Black-Right-Pointing-Pointer Sulfate-reducing bacteria and anode-respiring bacteria were enriched in anodic biofilms. Black-Right-Pointing-Pointer The MFC effectively remove sulfate to elementary sulfur in the presence of lactate. Black-Right-Pointing-Pointer The present device can treat sulfate laden wastewaters with electricity harvesting. - Abstract: Anaerobic treatment of sulfate-laden wastewaters can produce excess sulfide, which is corrosive to pipelines and is toxic to incorporated microorganisms. This work started up microbial fuel cell (MFC) using enriched sulfate-reducing mixed culture as anodic biofilms and applied the so yielded MFC for treating sulfate or sulfide-laden wastewaters. The sulfate-reducing bacteria in anodic biofilm effectively reduced sulfate to sulfide, which was then used by neighboring anode respiring bacteria (ARB) as electron donor for electricity production. The presence of organic carbons enhanced MFC performance since the biofilm ARB were mixotrophs that need organic carbon to grow. The present device introduces a route for treating sulfate laden wastewaters with electricity harvesting.

  1. Identification of triclosan-degrading bacteria in a triclosan enrichment culture using stable isotope probing.

    Science.gov (United States)

    Lee, Do Gyun; Cho, Kun-Ching; Chu, Kung-Hui

    2014-02-01

    Triclosan, a widely used antimicrobial agent, is an emerging contaminant in the environment. Despite its antimicrobial character, biodegradation of triclosan has been observed in pure cultures, soils and activated sludge. However, little is known about the microorganisms responsible for the degradation in mixed cultures. In this study, active triclosan degraders in a triclosan-degrading enrichment culture were identified using stable isotope probing (SIP) with universally (13)C-labeled triclosan. Eleven clones contributed from active microorganisms capable of uptake the (13)C in triclosan were identified. None of these clones were similar to known triclosan-degraders/utilizers. These clones distributed among α-, β-, or γ-Proteobacteria: one belonging to Defluvibacter (α-Proteobacteria), seven belonging to Alicycliphilus (β-Proteobacteria), and three belonging to Stenotrophomonas (γ-Proteobacteria). Successive additions of triclosan caused a significant shift in the microbial community structure of the enrichment culture, with dominant ribotypes belonging to the genera Alicycliphilus and Defluvibacter. Application of SIP has successfully identified diverse uncultivable triclosan-degrading microorganisms in an activated sludge enrichment culture. The results of this study not only contributed to our understanding of the microbial ecology of triclosan biodegradation in wastewater, but also suggested that triclosan degraders are more phylogenetically diverse than previously reported.

  2. Cholera Rapid Test with Enrichment Step Has Diagnostic Performance Equivalent to Culture

    Science.gov (United States)

    Ontweka, Lameck N.; Deng, Lul O.; Rauzier, Jean; Debes, Amanda K.; Tadesse, Fisseha; Parker, Lucy A.; Wamala, Joseph F.; Bior, Bior K.; Lasuba, Michael; But, Abiem Bona; Grandesso, Francesco; Jamet, Christine; Cohuet, Sandra; Ciglenecki, Iza; Serafini, Micaela; Sack, David A.; Quilici, Marie-Laure; Azman, Andrew S.; Luquero, Francisco J.

    2016-01-01

    Cholera rapid diagnostic tests (RDT) could play a central role in outbreak detection and surveillance in low-resource settings, but their modest performance has hindered their broad adoption. The addition of an enrichment step may improve test specificity. We describe the results of a prospective diagnostic evaluation of the Crystal VC RDT (Span Diagnostics, India) with enrichment step and of culture, each compared to polymerase chain reaction (PCR), during a cholera outbreak in South Sudan. RDTs were performed on alkaline peptone water inoculated with stool and incubated for 4–6 hours at ambient temperature. Cholera culture was performed from wet filter paper inoculated with stool. Molecular detection of Vibrio cholerae O1 by PCR was done from dry Whatman 903 filter papers inoculated with stool, and from wet filter paper supernatant. In August and September 2015, 101 consecutive suspected cholera cases were enrolled, of which 36 were confirmed by PCR. The enriched RDT had 86.1% (95% CI: 70.5–95.3) sensitivity and 100% (95% CI: 94.4–100) specificity compared to PCR as the reference standard. The sensitivity of culture versus PCR was 83.3% (95% CI: 67.2–93.6) for culture performed on site and 72.2% (95% CI: 54.8–85.8) at the international reference laboratory, where samples were tested after an average delay of two months after sample collection, and specificity was 98.5% (95% CI: 91.7–100) and 100% (95% CI: 94.5–100), respectively. The RDT with enrichment showed performance comparable to that of culture and could be a sustainable alternative to culture confirmation where laboratory capacity is limited. PMID:27992488

  3. Cholera Rapid Test with Enrichment Step Has Diagnostic Performance Equivalent to Culture.

    Science.gov (United States)

    Ontweka, Lameck N; Deng, Lul O; Rauzier, Jean; Debes, Amanda K; Tadesse, Fisseha; Parker, Lucy A; Wamala, Joseph F; Bior, Bior K; Lasuba, Michael; But, Abiem Bona; Grandesso, Francesco; Jamet, Christine; Cohuet, Sandra; Ciglenecki, Iza; Serafini, Micaela; Sack, David A; Quilici, Marie-Laure; Azman, Andrew S; Luquero, Francisco J; Page, Anne-Laure

    2016-01-01

    Cholera rapid diagnostic tests (RDT) could play a central role in outbreak detection and surveillance in low-resource settings, but their modest performance has hindered their broad adoption. The addition of an enrichment step may improve test specificity. We describe the results of a prospective diagnostic evaluation of the Crystal VC RDT (Span Diagnostics, India) with enrichment step and of culture, each compared to polymerase chain reaction (PCR), during a cholera outbreak in South Sudan. RDTs were performed on alkaline peptone water inoculated with stool and incubated for 4-6 hours at ambient temperature. Cholera culture was performed from wet filter paper inoculated with stool. Molecular detection of Vibrio cholerae O1 by PCR was done from dry Whatman 903 filter papers inoculated with stool, and from wet filter paper supernatant. In August and September 2015, 101 consecutive suspected cholera cases were enrolled, of which 36 were confirmed by PCR. The enriched RDT had 86.1% (95% CI: 70.5-95.3) sensitivity and 100% (95% CI: 94.4-100) specificity compared to PCR as the reference standard. The sensitivity of culture versus PCR was 83.3% (95% CI: 67.2-93.6) for culture performed on site and 72.2% (95% CI: 54.8-85.8) at the international reference laboratory, where samples were tested after an average delay of two months after sample collection, and specificity was 98.5% (95% CI: 91.7-100) and 100% (95% CI: 94.5-100), respectively. The RDT with enrichment showed performance comparable to that of culture and could be a sustainable alternative to culture confirmation where laboratory capacity is limited.

  4. 16S rRNA gene sequencing in routine identification of anaerobic bacteria isolated from blood cultures

    DEFF Research Database (Denmark)

    Justesen, Ulrik Stenz; Skov, Marianne Nielsine; Knudsen, Elisa;

    2010-01-01

    A comparison between conventional identification and 16S rRNA gene sequencing of anaerobic bacteria isolated from blood cultures in a routine setting was performed (n = 127). With sequencing, 89% were identified to the species level, versus 52% with conventional identification. The times...

  5. High frequency of Thermodesulfovibrio spp. and Anaerolineaceae in association with Methanoculleus spp. in a long-term incubation of n-alkanes-degrading methanogenic enrichment culture

    Directory of Open Access Journals (Sweden)

    Bo Liang

    2016-09-01

    Full Text Available In the present study, the microbial community and functional gene composition of a long-term active alkane-degrading methanogenic culture was established after two successive enrichment culture transfers and incubated for a total period of 1750 days. Molecular analysis was conducted after the second transfer (incubated for 750 days for both the active alkanes-degrading methanogenic enrichment cultures (T2-AE and the background control (T2-BC. A net increase of methane as the end product was detected in the headspace of the enrichment cultures amended with long-chain n-alkanes and intermediate metabolites, including octadecanoate, hexadecanoate, isocaprylate, butyrate, isobutyrate, propionate, acetate and formate were measured in the liquid cultures. The composition of microbial community shifted through the successive transfers over time of incubation. Sequences of bacterial and archaeal 16S rRNA gene (16S rDNA and mcrA functional gene indicated that bacterial sequences affiliated to Thermodesulfovibrio spp. and Anaerolineaceae and archaeal sequences falling within the genus Methanoculleus were the most frequently encountered and thus represented the dominant members performing the anaerobic degradation of long-chain n-alkanes and methanogenesis. In addition, the presence of assA functional genes encoding the alkylsuccinate synthase α subunit indicated that fumarate addition mechanism could be considered as a possible initial activation step of n-alkanes in the present study. The succession pattern of microbial communities indicates that Thermodesulfovibrio spp. could be a generalist participating in the metabolism of intermediates, while Anaerolineaceae plays a key role in the initial activation of long-chain n-alkane biodegradation.

  6. Developments in techniques for the isolation, enrichment, main culture conditions and identification of spermatogonial stem cells.

    Science.gov (United States)

    He, Yanan; Chen, Xiaoli; Zhu, Huabin; Wang, Dong

    2015-12-01

    The in vitro culture system of spermatogonial stem cells (SSCs) provides a basis for studies on spermatogenesis, and also contributes to the development of new methods for the preservation of livestock and animal genetic modification. In vitro culture systems have mainly been established for mouse SSCs, but are lacking for farm animals. We reviewed and analyzed the current progress in SSC techniques such as isolation, purification, cultivation and identification. Based on the published studies, we concluded that two-step enzyme digestion and magnetic-activated cell sorting are fast becoming the main methods for isolation and enrichment of SSCs. With regard to the culture systems, serum and feeders were earlier thought to play an important role in the self-renewal and proliferation of SSCs, but serum- and feeder-free culture systems as a means of overcoming the limitations of SSC differentiation in long-term SSC culture are being explored. However, there is still a need to establish more efficient and ideal culture systems that can also be used for SSC culture in larger mammals. Although the lack of SSC-specific surface markers has seriously affected the efficiency of purification and identification, the transgenic study is helpful for our identification of SSCs. Therefore, future studies on SSC techniques should focus on improving serum- and feeder-free culture techniques, and discovering and identifying specific surface markers of SSCs, which will provide new ideas for the optimization of SSC culture systems for mice and promote related studies in farm animals.

  7. Innovative Approaches Using Lichen Enriched Media to Improve Isolation and Culturability of Lichen Associated Bacteria

    Science.gov (United States)

    Biosca, Elena G.; Flores, Raquel; Santander, Ricardo D.; Díez-Gil, José Luis; Barreno, Eva

    2016-01-01

    Lichens, self-supporting mutualistic associations between a fungal partner and one or more photosynthetic partners, also harbor non-photosynthetic bacteria. The diversity and contribution of these bacteria to the functioning of lichen symbiosis have recently begun to be studied, often by culture-independent techniques due to difficulties in their isolation and culture. However, culturing as yet unculturable lichenic bacteria is critical to unravel their potential functional roles in lichen symbiogenesis, to explore and exploit their biotechnological potential and for the description of new taxa. Our objective was to improve the recovery of lichen associated bacteria by developing novel isolation and culture approaches, initially using the lichen Pseudevernia furfuracea. We evaluated the effect of newly developed media enriched with novel lichen extracts, as well as the influence of thalli washing time and different disinfection and processing protocols of thalli. The developed methodology included: i) the use of lichen enriched media to mimic lichen nutrients, supplemented with the fungicide natamycin; ii) an extended washing of thalli to increase the recovery of ectolichenic bacteria, thus allowing the disinfection of thalli to be discarded, hence enhancing endolichenic bacteria recovery; and iii) the use of an antioxidant buffer to prevent or reduce oxidative stress during thalli disruption. The optimized methodology allowed significant increases in the number and diversity of culturable bacteria associated with P. furfuracea, and it was also successfully applied to the lichens Ramalina farinacea and Parmotrema pseudotinctorum. Furthermore, we provide, for the first time, data on the abundance of culturable ecto- and endolichenic bacteria that naturally colonize P. furfuracea, R. farinacea and P. pseudotinctorum, some of which were only able to grow on lichen enriched media. This innovative methodology is also applicable to other microorganisms inhabiting these

  8. Effects of polychlorinated biphenyl congener concentration and sediment supplementation on rates of methanogenesis and 2,3,6-trichlorobiphenyl dechlorination in an anaerobic enrichment

    Energy Technology Data Exchange (ETDEWEB)

    Boyle, A.W.; May, H.D. (Celgene Corp., Warren, NJ (United States)); Blake, C.K. (Medical Univ. of South Carolina, Charleston, SC (United States)); Price, W.A. (Princeton Research Center, Lawrenceville, NJ (United States))

    1993-09-01

    The release of polychlorinated biphenyls (PCBs) into the environment has caused public concern. PCBs are know known to be susceptible to biodegradation; PCBs in the Hudson river have been shown to be extensively dechlorinated, but the rate of dechlorination in anaerobic environments have been slow, over months or years. This study tested the effects of PCB concentration and sediment supplementation with 2,3,6-trichlorobiphenol, on the rate of PCB dechlorination and methanogenesis. The rates of meta dechlorination in sediment supplemented cultures were measured in the laboratory, including both the rate per bacterial cell and the rate of methanogenesis when dechlorination increases. 24 refs., 3 figs., 1 tab.

  9. Isolation of Cultured Endothelial Progenitor Cells in vitro from PBMCs and CD133~+ Enriched Cells

    Institute of Scientific and Technical Information of China (English)

    郑伟红; 万亚峰; 马小鹏; 李兴睿; 杨志芳; 殷茜; 易继林

    2010-01-01

    Two isolation methods for sorting of endothelial progenitor cells(EPCs):from peripheral blood mononuclear cells(PBMCs)and CD133+ enriched cells were compared,by defining the cell morphology,phenotype,reproductive activities and function in vitro,to provide a reference for clinical application of EPCs.PBMCs from healthy subjects were used either directly for cell culture or for CD133+ sorting.The two groups of cells were cultured in complete medium 199(M199)for 7 to 14 days and the phenotypes of EPCs were an...

  10. Anaerobic bacteria grow within Candida albicans biofilms and induce biofilm formation in suspension cultures.

    Science.gov (United States)

    Fox, Emily P; Cowley, Elise S; Nobile, Clarissa J; Hartooni, Nairi; Newman, Dianne K; Johnson, Alexander D

    2014-10-20

    The human microbiome contains diverse microorganisms, which share and compete for the same environmental niches. A major microbial growth form in the human body is the biofilm state, where tightly packed bacterial, archaeal, and fungal cells must cooperate and/or compete for resources in order to survive. We examined mixed biofilms composed of the major fungal species of the gut microbiome, Candida albicans, and each of five prevalent bacterial gastrointestinal inhabitants: Bacteroides fragilis, Clostridium perfringens, Escherichia coli, Klebsiella pneumoniae, and Enterococcus faecalis. We observed that biofilms formed by C. albicans provide a hypoxic microenvironment that supports the growth of two anaerobic bacteria, even when cultured in ambient oxic conditions that are normally toxic to the bacteria. We also found that coculture with bacteria in biofilms induces massive gene expression changes in C. albicans, including upregulation of WOR1, which encodes a transcription regulator that controls a phenotypic switch in C. albicans, from the "white" cell type to the "opaque" cell type. Finally, we observed that in suspension cultures, C. perfringens induces aggregation of C. albicans into "mini-biofilms," which allow C. perfringens cells to survive in a normally toxic environment. This work indicates that bacteria and C. albicans interactions modulate the local chemistry of their environment in multiple ways to create niches favorable to their growth and survival.

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

    Science.gov (United States)

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

    2015-09-01

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

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

    Science.gov (United States)

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

    2015-01-01

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

  13. Microbial community analysis in sludge of anaerobic wastewater treatment systems : integrated culture-dependent and culture-independent approaches

    OpenAIRE

    Roest, C.

    2007-01-01

    The need for clean water is increasing and anaerobic wastewater treatment can be used as a cost-effective solution for purification of organically polluted industrial waste streams. This thesis presents results from microbiological investigations of several full-scale and lab-scale anaerobic wastewater treatments systems. Anaerobic wastewater treatment has gained popularity and is now one of the key technologies in environmental biotechnology. However, knowledge of the microbial community str...

  14. Bacterial community analysis of cypermethrin enrichment cultures and bioremediation of cypermethrin contaminated soils.

    Science.gov (United States)

    Akbar, Shamsa; Sultan, Sikander; Kertesz, Michael

    2015-07-01

    Cypermethrin is widely used for insect control; however, its toxicity toward aquatic life requires its complete removal from contaminated areas where the natural degradation ability of microbes can be utilized. Agricultural soil with extensive history of CM application was used to prepare enrichment cultures using cypermethrin as sole carbon source for isolation of cypermethrin degrading bacteria and bacterial community analysis using PCR-DGGE of 16 S rRNA gene. DGGE analysis revealed that dominant members of CM enrichment culture were associated with α-proteobacteria followed by γ-proteobacteria, Firmicutes, and Actinobacteria. Three potential CM-degrading isolates identified as Ochrobactrum anthropi JCm1, Bacillus megaterium JCm2, and Rhodococcus sp. JCm5 degraded 86-100% of CM (100 mg L(-1) ) within 10 days. These isolates were also able to degrade other pyrethroids, carbofuran, and cypermethrin degradation products. Enzyme activity assays revealed that enzymes involved in CM-degradation were inducible and showed activity when strains were grown on cypermethrin. Degradation kinetics of cypermethrin (200 mg kg(-1)) in soils inoculated with isolates JCm1, JCm2, and JCm5 suggested time-dependent disappearance of cypermethrin with rate constants of 0.0516, 0.0425, and 0.0807 d(-1), respectively, following first order rate kinetics. The isolated bacterial strains were among dominant genera selected under CM enriched conditions and represent valuable candidates for in situ bioremediation of contaminated soils and waters.

  15. Efficient anaerobic treatment of synthetic textile wastewater in a UASB reactor with granular sludge enriched with humic acids supported on alumina nanoparticles.

    Science.gov (United States)

    Cervantes, Francisco J; Gómez, Rafael; Alvarez, Luis H; Martinez, Claudia M; Hernandez-Montoya, Virginia

    2015-07-01

    A novel technique to co-immobilize humus-reducing microorganisms and humic substances (HS), supported on γ-Al2O3 nanoparticles (NP), by a granulation process in an upflow anaerobic sludge bed (UASB) reactor is reported in the present work. Larger granules (predominantly between 1 and 1.7 mm) were produced using NP coated with HS compared to those obtained with uncoated NP (mostly between 0.25 and 0.5 mm). The HS-enriched granular biomass was then tested for its capacity to achieve the reductive decolorization of the recalcitrant azo dye, reactive red 2 (RR2), in the same UASB reactor operated with a hydraulic residence time of 12 h and with glucose as electron donor. HS-enriched granules achieved higher decolorization and COD removal efficiencies, as compared to the control reactor operated in the absence of HS, in long term operation and applying high concentrations of RR2 (40-400 mg/L). This co-immobilizing technique could be attractive for its application in UASB reactors for the reductive biotransformation of several contaminants, such as nitroaromatics, poly-halogenated compounds, metalloids, among others.

  16. Microbial community analysis in sludge of anaerobic wastewater treatment systems : integrated culture-dependent and culture-independent approaches

    NARCIS (Netherlands)

    Roest, C.

    2007-01-01

    The need for clean water is increasing and anaerobic wastewater treatment can be used as a cost-effective solution for purification of organically polluted industrial waste streams. This thesis presents results from microbiological investigations of several full-scale and lab-scale anaerobic wastewa

  17. Improving the stability of thermophilic anaerobic digesters treating SS-OFMSW through enrichment with compost and leachate seeds

    KAUST Repository

    Ghanimeh, Sophia A.

    2013-03-01

    This paper examines the potential of improving the stability of thermophilic anaerobic digestion of source-sorted organic fraction of municipal solid waste (SS-OFMSW) by adding leachate and compost during inoculation. For this purpose, two stable thermophilic digesters, A (control) and B (with added leachate and compost), were subjected to a sustained substrate shock by doubling the organic loading rate for one week. Feeding was suspended then gradually resumed to reach the pre-shock loading rate (2. gVS/l/d). Digester A failed, exhibiting excessive increase in acetate and a corresponding decrease in pH and methane generation, and lower COD and solids removal efficiencies. In contrast, digester B was able to restore its functionality with 90% recovery of pre-shock methane generation rate at stable pH, lower hydrogen levels, and reduced VFAs and ammonia accumulation. © 2012 Elsevier Ltd.

  18. Inhibitory effects of butyrate on biological hydrogen production with mixed anaerobic cultures.

    Science.gov (United States)

    Zheng, Xian-Jun; Yu, Han-Qing

    2005-01-01

    In this study batch experiments were conducted to investigate the inhibitory effects of butyrate addition on hydrogen production from glucose by using anaerobic mixed cultures. Experimental results showed that addition of butyrate at 4.18 and 6.27 g/l only slightly inhibited hydrogen production, and addition of butyrate at 8.36-12.54 g/l imposed a moderate inhibitory effect on hydrogen production. At addition of 25.08 g/l, butyrate had a strong inhibitory influence on substrate degradation and hydrogen production. The distribution of the volatile fatty acids produced from the acidogeneisis of glucose was significantly influenced by the addition of butyrate. The inhibition of butyrate addition on hydrogen production was described well by a non-competitive and non-linear inhibition model, with the maximum hydrogen production rate of 59.3 ml/g-SS/h, critical added butyrate concentration of 25.08 g/l, and inhibition degree of 0.323, respectively. The C(I,50) values (the butyrate concentration at which bioactivity is reduced by 50%) for hydrogen production rate and yield were estimated as 19.39 and 20.78 g/l of added butyrate, respectively.

  19. Spirulina sp. LEB-18 culture using effluent from the anaerobic digestion

    Directory of Open Access Journals (Sweden)

    J. A. Borges

    2013-06-01

    Full Text Available The carbon source is the most expensive nutrient for Spirulina production; effluents from anaerobic digestion contain this nutrient in the form of HCO3-. The aim of this study was to assess the growth kinetics, composition and fatty acid profile of Spirulina sp. LEB-18 grown in standard Zarrouk medium (NaHCO3 16.8 g L-1 and in Zarrouk medium replaced with 20% (v/v effluent with reduced concentrations of NaHCO3 (5.3 and 2.8 g L-1. The use of effluent and lower concentrations of HCO3 was found to be an alternative to reduce the costs of Spirulina production, because there were no significant differences in growth parameters (µmax 0.324 - 0.354 d-1; Pmax 0.280 - 0.297 g L-1 d-1, in the different culture medium used. Lipids ranged between 4.9 and 5.0%; the media with effluent had higher levels of linoleic acid compared to the standard medium.

  20. Bio-Hydrogen Production from Pineapple Waste Extract by Anaerobic Mixed Cultures

    Directory of Open Access Journals (Sweden)

    Chakkrit Sreela-or

    2013-04-01

    Full Text Available A statistical experimental design was employed to optimize factors that affect the production of hydrogen from the glucose contained in pineapple waste extract by anaerobic mixed cultures. Results from Plackett-Burman design indicated that substrate concentration, initial pH and FeSO4 concentration had a statistically significant (p ≤ 0.05 influence on the hydrogen production potential (Ps and the specific hydrogen production rate (SHPR. The path of steepest ascent was undertaken to approach the optimal region of these three significant factors which was then optimized using response surface methodology (RSM with central composite design (CCD. The presence of a substrate concentration of 25.76 g-total sugar/L, initial pH of 5.56, and FeSO4 concentration of 0.81 g/L gave a maximum predicted Ps of 5489 mL H2/L, hydrogen yield of 1.83 mol H2/mol glucose, and SHPR of 77.31 mL H2/g-volatile suspended solid (VSS h. A verification experiment indicated highly reproducible results with the observed Ps and SHPR being only 1.13% and 1.14% different from the predicted values.

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

    DEFF Research Database (Denmark)

    De Abreu, Angela Alexandra Valente; Karakashev, Dimitar Borisov; Angelidaki, Irini;

    2012-01-01

    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...... 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....... Results Conversion of arabinose and glucose to hydrogen, by extreme thermophilic anaerobic mixed cultures was studied in continuous (70oC, pH 5.5) and batch (70oC, pH 5.5 and pH 7) assays. Two EGSB reactors, Rarab and Rgluc, were continuously fed with arabinose and glucose, respectively. No significant...

  2. Production of cellulases and hemicellulases by an anaerobic mixed culture from lignocellulosic biomass.

    Science.gov (United States)

    Tabassum, R; Rajoka, M I; Malik, K A

    1990-03-01

    A comparison of different habitats, biogas plant, rumen fluid and sewage sludge, for cellulolytic organisms indicated sewage studge was the best source. Enrichment cultura gave a mixed culture which exhibited CMCase activity as well as extracellular Avicelase, xylanase, β-glucosidase, β-xylosidase activities and cell-bound β-glucosidase, and β-xylosidase production in a synthetic medium with eleven different cellulosic and lignocellulosic substrates. The activity of extracellular β-glucosidase and β-xylosidase production was significantly higher than endogenous activities. Hemicellulases were induced better than cellulases. The anzyme system was stable under aerobic conditions. Of the different lignocellulosic substrates, kallar grass was the best inducer of extracellular enzymes.

  3. Identification of novel glycosyl hydrolases with cellulolytic activity against crystalline cellulose from metagenomic libraries constructed from bacterial enrichment cultures.

    Science.gov (United States)

    Mori, Toshio; Kamei, Ichiro; Hirai, Hirofumi; Kondo, Ryuichiro

    2014-01-01

    To obtain cellulases that are capable of degrading crystalline cellulose and cedar wood, metagenomic libraries were constructed from raw soil sample which was covered to pile of cedar wood sawdust or from its enrichment cultures. The efficiency of screening of metagenomic library was improved more than 3 times by repeating enrichment cultivation using crystalline cellulose as a carbon source, compared with the library constructed from raw soil. Four cellulase genes were obtained from the metagenomic libraries that were constructed from the total genome extracted from an enrichment culture that used crystalline cellulose as a carbon source. A cellulase gene and a xylanase gene were obtained from the enrichment culture that used unbleached kraft pulp as a carbon source. The culture supernatants of Escherichia coli expressing three clones that were derived from the enrichment culture that used crystalline cellulose showed activity against crystalline cellulose. In addition, these three enzyme solutions generated a reducing sugar from cedar wood powder. From these results, the construction of a metagenomic library from cultures that were repetition enriched using crystalline cellulose demonstrated that this technique is a powerful tool for obtaining cellulases that have activity toward crystalline cellulose.

  4. Detection of periodontopathogenic bacteria in pregnant women by traditional anaerobic culture method and by a commercial molecular genetic method.

    Science.gov (United States)

    Urbán, Edit; Terhes, Gabriella; Radnai, Márta; Gorzó, István; Nagy, Elisabeth

    2010-06-01

    To culture facultative and strict anaerobic bacteria is a well-established method for analyzing subgingival plaque samples. Micro-IDent and micro-IDent Plus (HAIN Lifescience GmbH, Nehren, Germany) tests are two commercially available rapid PCR-based methods for the identification and quantification of putative periodontopathogen bacteria. In this study, we compared these commercial PCR-based hybridization methods with conventional anaerobic culture technique. A total of 36 subgingival plaque samples were collected from periodontal pockets of pregnant women with chronic localized periodontitis. Aliquots of these samples were evaluated with species-specific probes provided by micro-IDent and micro-IDent Plus tests simultaneously, and from the same samples anaerobic and capnophylic bacteria were cultured on selective media. The overall agreement between both methods was excellent for Eubacterium nodatum, Tannerella forsythia and Porphyromonas gingivalis (97-92%), fair for Capnocytophaga sp, Eikenella corrodens, Actinobacillus actinomycetemcomitans, and Prevotella intermedia (91-89%) and poor for Fusobacterium nucleatum, Parvimonas micra (Micromonas micros), and Campylobacter rectus (86-78%). Discrepancies in the results may be explained by inability of culture method to distinguish between closely related taxa (e.i P. intermedia/Prevotella. nigrescens), and problems of keeping periodontopathogen bacteria viable, which is required for successful detection by standard culture method. Nucleic acid-based methods may replace cultivation method as frequently used methods in microbiological diagnosis of progressive periodontitis, thus micro-IDent and micro-IDent Plus tests can be recommended where culture of periodontopathogenic bacteria is not performed in routine microbiology laboratories to analyze subgingival plaque samples.

  5. Neonicotinoid Insecticides Alter the Gene Expression Profile of Neuron-Enriched Cultures from Neonatal Rat Cerebellum

    Directory of Open Access Journals (Sweden)

    Junko Kimura-Kuroda

    2016-10-01

    Full Text Available Neonicotinoids are considered safe because of their low affinities to mammalian nicotinic acetylcholine receptors (nAChRs relative to insect nAChRs. However, because of importance of nAChRs in mammalian brain development, there remains a need to establish the safety of chronic neonicotinoid exposures with regards to children’s health. Here we examined the effects of longterm (14 days and low dose (1 μM exposure of neuron-enriched cultures from neonatal rat cerebellum to nicotine and two neonicotinoids: acetamiprid and imidacloprid. Immunocytochemistry revealed no differences in the number or morphology of immature neurons or glial cells in any group versus untreated control cultures. However, a slight disturbance in Purkinje cell dendritic arborization was observed in the exposed cultures. Next we performed transcriptome analysis on total RNAs using microarrays, and identified significant differential expression (p < 0.05, q < 0.05, ≥1.5 fold between control cultures versus nicotine-, acetamiprid-, or imidacloprid-exposed cultures in 34, 48, and 67 genes, respectively. Common to all exposed groups were nine genes essential for neurodevelopment, suggesting that chronic neonicotinoid exposure alters the transcriptome of the developing mammalian brain in a similar way to nicotine exposure. Our results highlight the need for further careful investigations into the effects of neonicotinoids in the developing mammalian brain.

  6. Value of anaerobic blood culture in 4018 blood cultures samples%4018份血培养中厌氧血培养的价值分析

    Institute of Scientific and Technical Information of China (English)

    马艳; 胡必杰; 周春妹; 高晓东; 谢红梅; 黄声雷; 周昭彦; 鲍容

    2012-01-01

    目的 了解送检厌氧血培养瓶对病原菌检出率及阳性结果报告时间的影响.方法 对2011年1月-2012年3月送检的4018份疑似血流感染患者的血培养结果进行统计学分析.结果 同时送检需氧瓶和厌氧瓶的检出率达14.11%,高于仅送检需氧瓶的9.26%,差异有统计学意义(P<0.05);厌氧瓶中大肠埃希菌和肠球菌属的阳性结果报告时间(306、630min)明显短于需氧瓶(612、810 min)(P<0.05);同时送检需氧瓶和厌氧瓶的病份中,厌氧瓶阳性而需氧瓶阴性者占2.42%,厌氧瓶培养可增加血流感染病原菌检出率达17.11%.结论 增加厌氧瓶培养可以提高阳性率并缩短阳性结果报告时间,临床上要加强厌氧血培养瓶的送检.%OBJECTIVE To evaluate the clinical significance of anaerobic blood culture bottles in the detection rate of the pathogens and the time of positive reports. METHODS The blood culture was statistically analyzed for 4018 patients with suspected blood stream infections submitted from Jan 21011 to Mar 2012. RESULTS The detection rate of the aerobic bottles and anaerobic bottles submitted at the same period was 14. 11%. only higher than 9. 265-6 of the aerobic bottles, the difference was statistically significant (P<0. 05) ; on detecting Escherichia coli and Enterocaccus , anaerobic blood cultures bottles (306,630 min)needed significantly shorter time than did the aerobic blood botiles(612.810 min), (P<0. 05) ; 2. 42% of the cases only were determined positive in anaerobic blood cultures bottles while both aerobic and anaerobic bottles were obtained, the anaerobic blood cultures could increase 17. 11% of the isolation rate of the pathogens causing blood stream infections. CONCLUSION To increase the anaerobic bottle blood cultures may significantly increase the isolation rate and shorten the report time, it is necessary for the hospital to intensify the submission of the anaerobic blood culture bottles.

  7. Use of Response Surface Methodology to Optimize Culture Conditions for Hydrogen Production by an Anaerobic Bacterial Strain from Soluble Starch

    Science.gov (United States)

    Kieu, Hoa Thi Quynh; Nguyen, Yen Thi; Dang, Yen Thi; Nguyen, Binh Thanh

    2016-05-01

    Biohydrogen is a clean source of energy that produces no harmful byproducts during combustion, being a potential sustainable energy carrier for the future. Therefore, biohydrogen produced by anaerobic bacteria via dark fermentation has attracted attention worldwide as a renewable energy source. However, the hydrogen production capability of these bacteria depends on major factors such as substrate, iron-containing hydrogenase, reduction agent, pH, and temperature. In this study, the response surface methodology (RSM) with central composite design (CCD) was employed to improve the hydrogen production by an anaerobic bacterial strain isolated from animal waste in Phu Linh, Soc Son, Vietnam (PL strain). The hydrogen production process was investigated as a function of three critical factors: soluble starch concentration (8 g L-1 to 12 g L-1), ferrous iron concentration (100 mg L-1 to 200 mg L-1), and l-cysteine concentration (300 mg L-1 to 500 mg L-1). RSM analysis showed that all three factors significantly influenced hydrogen production. Among them, the ferrous iron concentration presented the greatest influence. The optimum hydrogen concentration of 1030 mL L-1 medium was obtained with 10 g L-1 soluble starch, 150 mg L-1 ferrous iron, and 400 mg L-1 l-cysteine after 48 h of anaerobic fermentation. The hydrogen concentration produced by the PL strain was doubled after using RSM. The obtained results indicate that RSM with CCD can be used as a technique to optimize culture conditions for enhancement of hydrogen production by the selected anaerobic bacterial strain. Hydrogen production from low-cost organic substrates such as soluble starch using anaerobic fermentation methods may be one of the most promising approaches.

  8. Anaerobic biodegradation of benzene series compounds by mixed cultures based on optional electronic acceptors

    Institute of Scientific and Technical Information of China (English)

    2007-01-01

    A series of batch experiments were performed using mixed bacterial consortia to investigate biodegradation performance of benzene,toluene,ethylbenzene and three xylene isomers (BTEX) under nitrate,sulfate and ferric iron reducing conditions.The results showed that toluene,ethylbenzeoe,m-xylene and o-xylene could be degraded independently by the mixed cultures coupled to nitrate,sulfate and ferric iron reduction.Under ferric iron reducing conditions the biodegradation of benzene and p-xylene could be occurred only in the presence of other alkylbenzenes.Alkylbenzenes can serve as the primary substrates to stimulate the transformation of benzene and p-xylene under anaerobic conditions.Benzene and p-xylene are more toxic than toluene and ethylbenzene,under the three terminal electron acceptors conditions,the degradation rates decreased with toluene > ethylbenzene > m-xylene > o-xylene > benzene > p-xylene.Nitrate was a more favorable electron acceptor compared to sulfate and ferric iron.The ratio between sulfate consumed and the loss of benzene,toluene,ethylbenzene,o-xylene,m-xylene,p-xylene was 4.44,4.51,4.42,4.32,4.37 and 4.23,respectively;the ratio between nitrate consumed and the loss of these substrates was 7.53,6.24,6.49,7.28,7.81,7.61,respectively;the ratio between the consumption of ferric iron and the loss of toluene,ethylbenzene,o-xylene,m-xylenewas 17.99,18.04,18.07,17.97,respectively.

  9. Regulation of Liver Enriched Transcription Factors in Rat Hepatocytes Cultures on Collagen and EHS Sarcoma Matrices.

    Directory of Open Access Journals (Sweden)

    Jürgen Borlak

    Full Text Available Liver-enriched transcription factors (LETF play a crucial role in the control of liver-specific gene expression and for hepatocytes to retain their molecular and cellular functions complex interactions with extra cellular matrix (ECM are required However, during cell isolation ECM interactions are disrupted and for hepatocytes to regain metabolic competency cells are cultured on ECM substrata. The regulation of LETFs in hepatocytes cultured on different ECM has not been studied in detail. We therefore compared two common sources of ECM and evaluated cellular morphology and hepatocyte differentiation by investigating DNA binding activity of LETFs at gene specific promoters and marker genes of hepatic metabolism. Furthermore, we studied testosterone metabolism and albumin synthesis to assess the metabolic competence of cell cultures. Despite significant difference in morphological appearance and except for HNF1β (p<0.001 most LETFs and several of their target genes did not differ in transcript expression after Bonferroni adjustment when cultured on collagen or Matrigel. Nonetheless, Western blotting revealed HNF1β, HNF3α, HNF3γ, HNF4α, HNF6 and the smaller subunits of C/EBPα and C/EBPβ to be more abundant on Matrigel cultured cells. Likewise, DNA binding activity of HNF3α, HNF3β, HNF4α, HNF6 and gene expression of hepatic lineage markers were increased on Matrigel cultured hepatocytes. To further investigate hepatic gene regulation, the effects of Aroclor 1254 treatment, e.g. a potent inducer of xenobiotic defense were studied in vivo and in vitro. The gene expression of C/EBP-α increased in rat liver and hepatocytes cultured on collagen and this treatment induced DNA binding activity of HNF4α, C/EBPα and C/EBPβ and gene expression of CYP1A1 and CYP1A2 in vivo and in vitro. Taken collectively, two sources of ECM greatly affected hepatocyte morphology, activity of liver enriched transcription factors, hepatic gene expression and

  10. Phylogenetic analysis of aerobic freshwater and marine enrichment cultures efficient in hydrocarbon degradation: effect of profiling method

    NARCIS (Netherlands)

    Chang, Y.J.; Stephen, J.R.; Richter, A.P.; Venosa, A.D.; Bruggemann, J.; MacNaughton, S.J.; Kowalchuk, G.A.; Haines, J.R.; Kline, E.; White, D.C.

    2000-01-01

    Aerobically grown enrichment cultures derived from hydrocarbon- contaminated seawater and freshwater sediments were generated by growth on crude oil as sole carbon source. Both cultures displayed a high rate of degradation for a wide range of hydrocarbon compounds. The bacterial species composition

  11. Impact of arachidonic acid enrichment of live rotifer prey on bacterial communities in rotifer and larval fish cultures.

    Science.gov (United States)

    Seychelles, Laurent H; Doiron, Kim; Audet, Céline; Tremblay, Réjean; Pernet, Fabrice; Lemarchand, Karine

    2013-03-01

    Rotifers (Brachionus plicatilis), commonly used at first feeding in commercial fish hatcheries, carry a large bacteria load. Because they are relatively poor in essential fatty acids, it is common practice to enrich them with fatty acids, including arachidonic acid (AA). This study aims to determine whether prey enrichment with AA may act as a prebiotic and modify the microbial community composition either in AA-enriched rotifer cultures or in larval-rearing water using winter flounder (Pseudopleuronectes americanus) as a larval fish model. AA enrichment modified the bacterial community composition in both the rotifer culture tanks and the larval-rearing tanks. We observed an increase in the number of cultivable bacteria on TCBS (thiosulfate-citrate-bile salts-sucrose) agar, used as a proxy for the abundance of Vibrio sp. The results suggest that AA may also play an indirect role in larval health.

  12. Persistence of pentolite (PETN and TNT) in soil microcosms and microbial enrichment cultures.

    Science.gov (United States)

    Arbeli, Ziv; Garcia-Bonilla, Erika; Pardo, Cindy; Hidalgo, Kelly; Velásquez, Trigal; Peña, Luis; C, Eliana Ramos; Avila-Arias, Helena; Molano-Gonzalez, Nicolás; Brandão, Pedro F B; Roldan, Fabio

    2016-05-01

    Pentolite is a mixture (1:1) of 2,4,6-trinitrotoluene (TNT) and pentaerythritol tetranitrate (PETN), and little is known about its fate in the environment. This study was aimed to determine the dissipation of pentolite in soils under laboratory conditions. Microcosm experiments conducted with two soils demonstrated that dissipation rate of PETN was significantly slower than that of TNT. Interestingly, the dissipation of PETN was enhanced by the presence of TNT, while PETN did not enhanced the dissipation of TNT. Pentolite dissipation rate was significantly faster under biostimulation treatment (addition of carbon source) in soil from the artificial wetland, while no such stimulation was observed in soil from detonation field. In addition, the dissipation rate of TNT and PETN in soil from artificial wetland under biostimulation was significantly faster than the equivalent abiotic control, although it seems that non-biological processes might also be important for the dissipation of TNT and PETN. Transformation of PETN was also slower during establishment of enrichment culture using pentolite as the sole nitrogen source. In addition, transformation of these explosives was gradually reduced and practically stopped after the forth cultures transfer (80 days). DGGE analysis of bacterial communities from these cultures indicates that all consortia were dominated by bacteria from the order Burkholderiales and Rhodanobacter. In conclusion, our results suggest that PETN might be more persistent than TNT.

  13. Selective enrichment and production of highly urease active bacteria by non-sterile (open) chemostat culture.

    Science.gov (United States)

    Cheng, Liang; Cord-Ruwisch, Ralf

    2013-10-01

    In general, bioprocesses can be subdivided into naturally occurring processes, not requiring sterility (e.g., beer brewing, wine making, lactic acid fermentation, or biogas digestion) and other processes (e.g., the production of enzymes and antibiotics) that typically require a high level of sterility to avoid contaminant microbes overgrowing the production strain. The current paper describes the sustainable, non-sterile production of an industrial enzyme using activated sludge as inoculum. By using selective conditions (high pH, high ammonia concentration, and presence of urea) for the target bacterium, highly active ureolytic bacteria, physiologically resembling Sporosarcina pasteurii were reproducibly enriched and then continuously produced via chemostat operation of the bioreactor. When using a pH of 10 and about 0.2 M urea in a yeast extract-based medium, ureolytic bacteria developed under aerobic chemostat operation at hydraulic retention times of about 10 h with urease levels of about 60 μmol min⁻¹ ml⁻¹ culture. For cost minimization at an industrial scale the costly protein-rich yeast extract medium could be replaced by commercial milk powder or by lysed activated sludge. Glutamate, molasses, or glucose-based media did not result in the enrichment of ureolytic bacteria by the chemostat. The concentration of intracellular urease was sufficiently high such that the produced raw effluent from the reactor could be used directly for biocementation in the field.

  14. Fermentative bio-hydrogen production from cellulose by cow dung compost enriched cultures

    Energy Technology Data Exchange (ETDEWEB)

    Ren, Nan-Qi; Xu, Ji-Fei; Gao, Ling-Fang; Xin, Liang; Qiu, Jie; Su, Dong-Xia [State Key Laboratory of Urban Water Resources and Environment, Harbin Institute of Technology, Harbin 150090 (China)

    2010-04-15

    The performance of hydrogen production from cellulose by the cow dung compost enriched continuously in defined medium containing cellulose was investigated. In the initial experiments, batch-fermentation was carried out to observe the effects of different substrate concentration conditions on the rate of cellulose-degrading, growth of bacteria and the capability of hydrogen-producing from cellulose. The result showed that the cellulose degradation decreased from 55% at 5 g/l to 22% at 30 g/l. The maximum cumulative hydrogen production and the rate of hydrogen production first increased from 828 ml/l at 5 g/l to 1251 ml/l at 10 g/l then remained constant beyond 10 g/l. The maximum hydrogen production potential, the rate of hydrogen production and the yield of hydrogen was 1525 ml/l, 33 ml/l.h, and 272 ml/g-cellulose (2.09 mol/mol-hexose) was obtained at substrate concentration 10 g/l, the hydrogen concentration in biogas was 47-50%(v/v) and there was no methane observed. During the conversion of cellulose into hydrogen, acetate and butyrate were main liquid end-products in the metabolism of hydrogen fermentation. These results proposed that cow dung compost enriched cultures were ideal microflora for hydrogen production from cellulose. (author)

  15. Detection of Yersinia spp. in meat products by enrichment culture, immunomagnetic separation and nested PCR.

    Science.gov (United States)

    Estrada, Cecilia S M Lucero; Velázquez, Lidia Del Carmen; Favier, Gabriela Isabel; Genaro, María Silvia Di; Escudero, María Esther

    2012-05-01

    The prevalence of Yersinia enterocolitica in meat products was assessed by four methods: cold enrichment in trypticase soy broth (A), enrichment in modified Rappaport broth at 25 °C (B), concentration by immunomagnetic separation (C) and yadA nested PCR (D). Furthermore, the pathogenic potentials of the isolates were established by phenotypic and genotypic tests, and their genomic relationships were determined by pulsed-field gel electrophoresis (PFGE). A total of 238 samples were collected at retail level in the city of San Luis, Argentina, during the period 2007-2008. The highest Yersinia prevalence in meat products was observed by method D (92 positive samples), followed by methods A (13 positive samples) and C (5 positive samples); however, no isolation was obtained by method B. Fourteen Y. enterocolitica and 4 Yersinia intermedia strains were recovered by culture. All Y. enterocolitica 2/O:9 strains gave results related to virulence by phenotypic tests and exhibited the genotype virF(+)myfA(+)ail(+)ystA(+). Two biotype 1A strains showed a genotype virF(-)myfA(-)ail(+)ystA(+)ystB(+). The 14 Y. enterocolitica strains isolated during this work plus one reference strain were separated into 11 genomic types by PFGE. This genomic heterogeneity of the isolates shows the diversity of Y. enterocolitica strains in our region. It is the first time that IMS was used to search Y. enterocolitica strains from naturally contaminated meat products.

  16. Dissimilatory Reduction of Elemental Selenium to Selenide in Sediments and Anaerobic Cultures of Selenium Respiring Bacteria

    Science.gov (United States)

    Herbel, M. J.; Switzer-Blum, J.; Oremland, R. S.

    2001-12-01

    Selenium contaminated environments often contain elemental Se (Se0) in their sediments that originates from dissimilatory reduction of Se oxyanions. The forms of Se in sedimentary rocks similarly contain high proportions of Se0, but much of the Se is also in the form of metal selenides, Se-2. It is not clear if the occurrence of these selenides is due to microbial reduction of Se0, or some other biological or chemical process. In this investigation we examined the possibility that bacterial respiratory reduction of Se0 to Se-2 could explain the presence of the latter species in sedimentary rocks. We conducted incubations of anoxic sediment slurries amended with different forms of Se0. High levels of Se0 (mM) were added to San Francisco Bay sediments in order to enhance the detection of soluble HSe-, which was precipitated with Cu2+ then redissolved and quantified by ICP-MS. Concentrations of HSe- were highest in live samples amended with red amorphous Se0 formed by either microbial reduction of Se+4 ("biogenic Se0") or by chemical oxidation of H2Se(g) ("chem. Se0"); very little HSe- was formed in those amended with black crystalline Se0, indicating the general lack of reactivity of this allotrope. Controls poisoned with 10% formalin did not produce HSe- from additions of chem. Se0. Reduction of both forms of red amorphous Se0 to HSe- occurred vigorously in growing cultures of Bacillus selenitireducens, an anaerobic halophile previously isolated from sediments of Mono Lake, CA. Up to 73% and 68% of red amorphous, biogenic Se0 or chem. Se0, respectively, was reduced to HSe- during growth of B. selenitireducens, (incubation time ~ 200 hrs): oxidation of lactate to acetate as well as cell density increases indicated that a dissimilatory reduction pathway was likely. Reduction was most enhanced when cells were previously grown on elemental sulfur or Se+4. In contrast to the growth experiments, washed cell suspensions of B. selenitireducens exhibited no HSe- production

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

    Directory of Open Access Journals (Sweden)

    Abreu Angela A

    2012-02-01

    Full Text Available Abstract 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

  18. Hydrogenotrophic culture enrichment reveals rumen Lachnospiraceae and Ruminococcaceae acetogens and hydrogen-responsive Bacteroidetes from pasture-fed cattle.

    Science.gov (United States)

    Gagen, Emma J; Padmanabha, Jagadish; Denman, Stuart E; McSweeney, Christopher S

    2015-07-01

    Molecular information suggests that there is a broad diversity of acetogens in the rumen, distinct from any currently isolated acetogens. We combined molecular analysis with enrichment culture techniques to investigate this diversity further. Methane-inhibited, hydrogenotrophic enrichment cultures produced acetate as the dominant end product. Acetyl-CoA synthase gene analysis revealed putative acetogens in the cultures affiliated with the Lachnospiraceae and Ruminococcaceae as has been found in other rumen studies. No formyltetrahydrofolate synthetase genes affiliating with acetogens or with 'homoacetogen similarity' scores >90% were identified. To further investigate the hydrogenotrophic populations in these cultures and link functional gene information with 16S rRNA gene identity, cultures were subcultured quickly, twice, through medium without exogenous hydrogen, followed by incubation without exogenous hydrogen. Comparison of cultures lacking hydrogen and their parent cultures revealed novel Lachnospiraceae and Ruminococcaceae that diminished in the absence of hydrogen, supporting the hypothesis that they were likely the predominant acetogens in the enrichments. Interestingly, a range of Bacteroidetes rrs sequences that demonstrated <86% identity to any named isolate also diminished in cultures lacking hydrogen. Acetogens or sulphate reducers from the Bacteroidetes have not been reported previously; therefore this observation requires further investigation.

  19. Therapy of Chronic Cardiosclerosis in WAG Rats Using Cultures of Cardiovascular Cells Enriched with Cardiac Stem Cell.

    Science.gov (United States)

    Chepeleva, E V; Pavlova, S V; Malakhova, A A; Milevskaya, E A; Rusakova, Ya L; Podkhvatilina, N A; Sergeevichev, D S; Pokushalov, E A; Karaskov, A M; Sukhikh, G T; Zakiyan, S M

    2015-11-01

    We developed a protocol for preparing cardiac cell culture from rat heart enriched with regional stem cells based on clonogenic properties and proliferation in culture in a medium with low serum content. Experiments on WAG rats with experimental ischemic myocardial damage showed that implantation of autologous regional stem cells into the left ventricle reduced the volume of cicatricial tissue, promoted angiogenesis in the damaged zone, and prevented the risk of heart failure development.

  20. High molecular weight dissolved organic matter enrichment selects for methylotrophs in dilution to extinction cultures.

    Science.gov (United States)

    Sosa, Oscar A; Gifford, Scott M; Repeta, Daniel J; DeLong, Edward F

    2015-12-01

    The role of bacterioplankton in the cycling of marine dissolved organic matter (DOM) is central to the carbon and energy balance in the ocean, yet there are few model organisms available to investigate the genes, metabolic pathways, and biochemical mechanisms involved in the degradation of this globally important carbon pool. To obtain microbial isolates capable of degrading semi-labile DOM for growth, we conducted dilution to extinction cultivation experiments using seawater enriched with high molecular weight (HMW) DOM. In total, 93 isolates were obtained. Amendments using HMW DOM to increase the dissolved organic carbon concentration 4x (280 μM) or 10x (700 μM) the ocean surface water concentrations yielded positive growth in 4-6% of replicate dilutions, whereas <1% scored positive for growth in non-DOM-amended controls. The majority (71%) of isolates displayed a distinct increase in cell yields when grown in increasing concentrations of HMW DOM. Whole-genome sequencing was used to screen the culture collection for purity and to determine the phylogenetic identity of the isolates. Eleven percent of the isolates belonged to the gammaproteobacteria including Alteromonadales (the SAR92 clade) and Vibrio. Surprisingly, 85% of isolates belonged to the methylotrophic OM43 clade of betaproteobacteria, bacteria thought to metabolically specialize in degrading C1 compounds. Growth of these isolates on methanol confirmed their methylotrophic phenotype. Our results indicate that dilution to extinction cultivation enriched with natural sources of organic substrates has a potential to reveal the previously unsuspected relationships between naturally occurring organic nutrients and the microorganisms that consume them.

  1. Enrichment and isolation of Bacillus beveridgei sp. nov., a facultative anaerobic haloalkaliphile from Mono Lake, California, that respires oxyanions of tellurium, selenium, and arsenic

    Science.gov (United States)

    Baesman, S.M.; Stolz, J.F.; Kulp, T.R.; Oremland, R.S.

    2009-01-01

    Mono Lake sediment slurries incubated with lactate and tellurite [Te(IV)] turned progressively black with time because of the precipitation of elemental tellurium [Te(0)]. An enrichment culture was established from these slurries that demonstrated Te(IV)-dependent growth. The enrichment was purified by picking isolated black colonies from lactate/Te(IV) agar plates, followed by repeated streaking and picking. The isolate, strain MLTeJB, grew in aqueous Te(IV)-medium if provided with a small amount of sterile solid phase material (e.g., agar plug; glass beads). Strain MLTeJB grew at high concentrations of Te(IV) (~8 mM) by oxidizing lactate to acetate plus formate, while reducing Te(IV) to Te(0). Other electron acceptors that were found to sustain growth were tellurate, selenate, selenite, arsenate, nitrate, nitrite, fumarate and oxygen. Notably, growth on arsenate, nitrate, nitrite and fumarate did not result in the accumulation of formate, implying that in these cases lactate was oxidized to acetate plus CO2. Strain MLTeJB is a low G + C Gram positive motile rod with pH, sodium, and temperature growth optima at 8.5-9.0, 0.5-1.5 M, and 40??C, respectively. The epithet Bacillus beveridgei strain MLTeJBT is proposed. ?? 2009 Springer.

  2. Anaerobic gut fungi: Advances in isolation, culture, and cellulolytic enzyme discovery for biofuel production.

    Science.gov (United States)

    Haitjema, Charles H; Solomon, Kevin V; Henske, John K; Theodorou, Michael K; O'Malley, Michelle A

    2014-08-01

    Anaerobic gut fungi are an early branching family of fungi that are commonly found in the digestive tract of ruminants and monogastric herbivores. It is becoming increasingly clear that they are the primary colonizers of ingested plant biomass, and that they significantly contribute to the decomposition of plant biomass into fermentable sugars. As such, anaerobic fungi harbor a rich reservoir of undiscovered cellulolytic enzymes and enzyme complexes that can potentially transform the conversion of lignocellulose into bioenergy products. Despite their unique evolutionary history and cellulolytic activity, few species have been isolated and studied in great detail. As a result, their life cycle, cellular physiology, genetics, and cellulolytic metabolism remain poorly understood compared to aerobic fungi. To help address this limitation, this review briefly summarizes the current body of knowledge pertaining to anaerobic fungal biology, and describes progress made in the isolation, cultivation, molecular characterization, and long-term preservation of these microbes. We also discuss recent cellulase- and cellulosome-discovery efforts from gut fungi, and how these interesting, non-model microbes could be further adapted for biotechnology applications.

  3. Lack of cardiac differentiation in c-kit-enriched porcine bone marrow and spleen hematopoietic cell cultures using 5-azacytidine

    NARCIS (Netherlands)

    M.L. Ramirez (Mario); T. McMorrow (Tara); T.M. Sanderson (Thomas M.); C.J. Lancos (C.); Y.-L. Tseng (Y.); D.K.C. Cooper (David); F.J.M.F. Dor (Frank)

    2005-01-01

    textabstractThe adult spleen is a source of early hematopoietic stem cells (HSC). We therefore studied whether culturing spleen or bone marrow (BM) HSC in medium containing 5-azacytidine could induce a cardiac phenotype. c-kit enrichment and depletion of adult pig spleen and BM mononuclear cells wer

  4. Feasibility study of an alkaline-based chemical treatment for the purification of polyhydroxybutyrate produced by a mixed enriched culture

    NARCIS (Netherlands)

    Jiang, Y.; Mikova, G.; Kleerebezem, R.; Van der Wielen, L.A.M.; Cuellar Soares, M.C.

    2015-01-01

    This study focused on investigating the feasibility of purifying polyhydroxybutyrate (PHB) from mixed culture biomass by alkaline-based chemical treatment. The PHB-containing biomass was enriched on acetate under non-sterile conditions. Alkaline treatment (0.2 M NaOH) together with surfactant SDS (0

  5. Stable acetate production in extreme-thermophilic (70ºC) mixed culture fermentation by selective enrichment of hydrogenotrophic methanogens

    NARCIS (Netherlands)

    Zhang, F.; Zhang, Y.; Ding, J.; Dai, K.; Van Loosdrecht, M.C.M.; Zeng, R.J.

    2014-01-01

    The control of metabolite production is difficult in mixed culture fermentation. This is particularly related to hydrogen inhibition. In this work, hydrogenotrophic methanogens were selectively enriched to reduce the hydrogen partial pressure and to realize efficient acetate production in extreme-th

  6. Timeline of bio-hydrogen production by anaerobic digestion of biomass

    OpenAIRE

    Bernadette E. TELEKY; Mugur C. BĂLAN; Nikolausz, Marcell

    2015-01-01

    Anaerobic digestion of biomass is a process capable to produce biohydrogen, a clean source of alternative energy. Lignocellulosic biomass from agricultural waste is considered a renewable energy source; therefore its utilization also contributes to the reduction of water, soil and air pollution. The study consists in five consecutive experiments designed to utilize anaerobic bacterial enrichment cultures originating from the Hungarian Lake, Hévíz. Wheat straw was used as com...

  7. Metagenomic and proteomic analyses to elucidate the mechanism of anaerobic benzene degradation

    Energy Technology Data Exchange (ETDEWEB)

    Abu Laban, Nidal [Helmholtz (Germany)

    2011-07-01

    This paper presents the mechanism of anaerobic benzene degradation using metagenomic and proteomic analyses. The objective of the study is to find out the microbes and biochemistry involved in benzene degradation. Hypotheses are proposed for the initial activation mechanism of benzene under anaerobic conditions. Two methods for degradation, molecular characterization and identification of benzene-degrading enzymes, are described. The physiological and molecular characteristics of iron-reducing enrichment culture are given and the process is detailed. Metagenome analysis of iron-reducing culture is presented using a pie chart. From the metagenome analysis of benzene-degrading culture, putative mobile element genes were identified in the aromatic-degrading configurations. Metaproteomic analysis of iron-reducing cultures and the anaerobic benzene degradation pathway are also elucidated. From the study, it can be concluded that gram-positive bacteria are involved in benzene degradation under iron-reducing conditions and that the catalysis mechanism of putative anaerobic benzene carboxylase needs further investigation.

  8. Influence of an aerobic fungus grown on solid culture on ruminal degradability and on a mixture culture of anaerobic cellulolytic bacteria.

    Science.gov (United States)

    Hernández-Díaz, R; Pimentel-González, D J; Figueira, A C; Viniegra-González, G; Campos-Montiel, R G

    2010-06-01

    In this work, the effect of a solid fungal culture of Aspergillus niger (An) grown on coffee pulp on the in situ ruminal degradability (RD) of corn stover was evaluated. In addition, the effect of its extracts on the in vitro dry matter disappearance (IVDMD) and on a mixed culture of anaerobic cellulolytic bacteria (MCACB) was also investigated. The solid ferment was a crude culture of An, grown on coffee pulp. Regarding in situ RD, a significant difference (p < 0.05) was found between treatment with 200 g/day of the solid culture and control (no solid culture added) on dry matter, crude protein and neutral detergent fibre on RD. All the water extracts (pH 4, 7 and 10) enhanced IVDMD and stimulated the cellulolytic activity on a MCACB. Ultrafiltration results showed that active compounds with a molecular weight lower than 30 kDa were responsible for the effect on MCACB. Such results suggest that the effects of the solid An culture in RD are related to the presence of water soluble compounds having a molecular weight lower than 30 kDa.

  9. Large-scale monocyte enrichment coupled with a closed culture system for the generation of human dendritic cells.

    Science.gov (United States)

    Pullarkat, Vinod; Lau, Roy; Lee, Sun-Min; Bender, James G; Weber, Jeffrey S

    2002-09-15

    Conventional methods for generating monocyte-derived dendritic cells (DC) for clinical trials utilize the property of plastic adherence to select monocytes from leukapheresis samples. This method is labor-intensive and has the potential for contamination at various steps. We evaluated a large-scale monocyte enrichment procedure using a cell selector (Isolex 300i(R)) followed by culture in a sterile bag system (Stericell(R)) for generation of DC. DC generated in tissue culture flasks after monocyte selection by plastic adherence were compared to those generated in Stericell(R) bags after monocyte enrichment by negative selection with the Isolex(R) 300i. DC were matured with lipopolysaccharide and pulsed with a peptide derived from the melanoma antigen gp100. Peptide-pulsed DC cultured by the two techniques were evaluated for phenotype, viability, ability to induce allogeneic and peptide-specific autologous proliferative responses as well as peptide-specific cytotoxic T-cell responses. The mean monocyte yield from leukapheresis collections was 17+/-2.4%, which increased to 52+/-11% after Isolex(R) selection. The DC yield of plated mononuclear cells from flasks or bags was 2.7+/-0.96% and 4.84+/-2.65%, respectively. DC cultured by both methods expressed high levels of CD86, CD80, CD40, CD83, CD44, CD11c and CD58, and was comparable in their ability to induce allogeneic and peptide-specific autologous proliferative responses as well as gp100 peptide-specific cytotoxic T-cell responses. These results indicate that potent monocyte-derived DC can be generated in a closed culture bag system after monocyte enrichment by immunomagnetic negative selection. Due to the closed nature of the enrichment and culture systems, the potential for contamination is minimized. This protocol is well suited for culturing large numbers of DC for clinical immunotherapy trials.

  10. The culture of cancer cell lines as tumorspheres does not systematically result in cancer stem cell enrichment.

    Science.gov (United States)

    Calvet, Christophe Y; André, Franck M; Mir, Lluis M

    2014-01-01

    Cancer stem cells (CSC) have raised great excitement during the last decade and are promising targets for an efficient treatment of tumors without relapses and metastases. Among the various methods that enable to enrich cancer cell lines in CSC, tumorspheres culture has been predominantly used. In this report, we attempted to generate tumorspheres from several murine and human cancer cell lines: B16-F10, HT-29, MCF-7 and MDA-MB-231 cells. Tumorspheres were obtained with variable efficiencies from all cell lines except from MDA-MB-231 cells. Then, we studied several CSC characteristics in both tumorspheres and adherent cultures of the B16-F10, HT-29 and MCF-7 cells. Unexpectedly, tumorspheres-forming cells were less clonogenic and, in the case of B16-F10, less proliferative than attached cells. In addition, we did not observe any enrichment in the population expressing CSC surface markers in tumorspheres from B16-F10 (CD133, CD44 and CD24 markers) or MCF-7 (CD44 and CD24 markers) cells. On the contrary, tumorspheres culture of HT-29 cells appeared to enrich in cells expressing colon CSC markers, i.e. CD133 and CD44 proteins. For the B16-F10 cell line, when 1 000 cells were injected in syngenic C57BL/6 mice, tumorspheres-forming cells displayed a significantly lower tumorigenic potential than adherent cells. Finally, tumorspheres culture of B16-F10 cells induced a down-regulation of vimentin which could explain, at least partially, the lower tumorigenicity of tumorspheres-forming cells. All these results, along with the literature, indicate that tumorspheres culture of cancer cell lines can induce an enrichment in CSC but in a cell line-dependent manner. In conclusion, extensive characterization of CSC properties in tumorspheres derived from any cancer cell line or cancer tissue must be performed in order to ensure that the generated tumorspheres are actually enriched in CSC.

  11. Biodegradation of Various Aromatic Compounds by Enriched Bacterial Cultures: Part A-Monocyclic and Polycyclic Aromatic Hydrocarbons.

    Science.gov (United States)

    Oberoi, Akashdeep Singh; Philip, Ligy; Bhallamudi, S Murty

    2015-08-01

    Present study focused on the screening of bacterial consortium for biodegradation of monocyclic aromatic hydrocarbon (MAH) and polycyclic aromatic hydrocarbons (PAHs). Target compounds in the present study were naphthalene, acenaphthene, phenanthrene (PAHs), and benzene (MAH). Microbial consortia enriched with the above target compounds were used in screening experiments. Naphthalene-enriched consortium was found to be the most efficient consortium, based on its substrate degradation rate and its ability to degrade other aromatic pollutants with significantly high efficiency. Substrate degradation rate with naphthalene-enriched culture followed the order benzene > naphthalene > acenaphthene > phenanthrene. Chryseobacterium and Rhodobacter were discerned as the predominant species in naphthalene-enriched culture. They are closely associated to the type strain Chryseobacterium arthrosphaerae and Rhodobacter maris, respectively. Single substrate biodegradation studies with naphthalene (PAH) and benzene (MAH) were carried out using naphthalene-enriched microbial consortium (NAPH). Phenol and 2-hydroxybenzaldehyde were identified as the predominant intermediates during benzene and naphthalene degradation, respectively. Biodegradation of toluene, ethyl benzene, xylene, phenol, and indole by NAPH was also investigated. Monod inhibition model was able to simulate biodegradation kinetics for benzene, whereas multiple substrate biodegradation model was able to simulate biodegradation kinetics for naphthalene.

  12. Isolation and characterization of a mesophilic heavy-metals-tolerant sulfate-reducing bacterium Desulfomicrobium sp. from an enrichment culture using phosphogypsum as a sulfate source.

    Science.gov (United States)

    Azabou, Samia; Mechichi, Tahar; Patel, Bharat K C; Sayadi, Sami

    2007-02-09

    A sulfate-reducing bacterium, was isolated from a 6 month trained enrichment culture in an anaerobic media containing phosphogypsum as a sulfate source, and, designated strain SA2. Cells of strain SA2 were rod-shaped, did not form spores and stained Gram-negative. Phylogenetic analysis of the 16S rRNA gene sequence of the isolate revealed that it was related to members of the genus Desulfomicrobium (average sequence similarity of 98%) with Desulfomicrobium baculatum being the most closely related (sequence similarity of 99%). Strain SA2 used thiosulfate, sulfate, sulfite and elemental sulfur as electron acceptors and produced sulfide. Strain SA2 reduced sulfate contained in 1-20g/L phosphogypsum to sulfide with reduction of sulfate contained in 2g/L phosphogypsum being the optimum concentration. Strain SA2 grew with metalloid, halogenated and non-metal ions present in phosphogypsum and with added high concentrations of heavy metals (125ppm Zn and 100ppm Ni, W, Li and Al). The relative order for the inhibitory metal concentrations, based on the IC(50) values, was Cu, Te>Cd>Fe, Co, Mn>F, Se>Ni, Al, Li>Zn.

  13. Transcriptional monitoring of steady state and effects of anaerobic phases in chemostat cultures of the filamentous fungus Trichoderma reesei

    Directory of Open Access Journals (Sweden)

    Penttilä Merja

    2006-10-01

    Full Text Available Abstract Background Chemostat cultures are commonly used in production of cellular material for systems-wide biological studies. We have used the novel TRAC (transcript analysis with aid of affinity capture method to study expression stability of approximately 30 process relevant marker genes in chemostat cultures of the filamentous fungus Trichoderma reesei and its transformant expressing laccase from Melanocarpus albomyces. Transcriptional responses caused by transient oxygen deprivations and production of foreign protein were also studied in T. reesei by TRAC. Results In cultures with good steady states, the expression of the marker genes varied less than 20% on average between sequential samples for at least 5 or 6 residence times. However, in a number of T. reesei cultures continuous flow did not result in a good steady state. Perturbations to the steady state were always evident at the transcriptional level, even when they were not measurable as changes in biomass or product concentrations. Both unintentional and intentional perturbations of the steady state demonstrated that a number of genes involved in growth, protein production and secretion are sensitive markers for culture disturbances. Exposure to anaerobic conditions caused strong responses at the level of gene expression, but surprisingly the cultures could regain their previous steady state quickly, even after 3 h O2 depletion. The main effect of producing M. albomyces laccase was down-regulation of the native cellulases compared with the host strain. Conclusion This study demonstrates the usefulness of transcriptional analysis by TRAC in ensuring the quality of chemostat cultures prior to costly and laborious genome-wide analysis. In addition TRAC was shown to be an efficient tool in studying gene expression dynamics in transient conditions.

  14. Survey of Naegleria from Taiwan recreational waters using culture enrichment combined with PCR.

    Science.gov (United States)

    Huang, Shih-Wei; Hsu, Bing-Mu

    2011-08-01

    Naegleria is a free-living amoeba. Pathogenic Naegleria may pose a health risk to people who come in contact with recreational waters. Here, we used Naegleria culture enrichment with PCR to identify the Naegleria species and investigated the distribution of Naegleria spp. in recreational waters including spring water, stream water and raw domestic water in central and southern Taiwan. In this study, Naegleria spp. were detected in 19 (17.8%) of the water samples. The occurrence of Naegleria in raw domestic water was 28.6%, higher than in stream water (14.7%) and in spring water (6.5%). The most frequently identified species exhibiting the closest phylogenetic relationships to the isolates were N. australiensis (n=4) and N. canariensis (n=4), followed by N. clarki (n=3) and N. philippinensis (n=3); N. americana (n=2). N. lovaniensis, N. dobsoni, and N. gruberi were each detected once. The pathogenic species N. fowleri was not detected, probably due to the low incubation temperature; however, the isolates exhibiting the closest phylogenetic relationships to the pathogenic species in mice of PAM, N. australiensis and N. philippinensis, were found. Results of this survey suggest the distribution of Naegleria spp. excluding N. fowleri in recreational waters. It should be considered a potential threat for health associated with human activities in recreational waters.

  15. Fermentative Hydrogen Production by Pure Culture with a New H2-producing Anaerobe

    Institute of Scientific and Technical Information of China (English)

    LI Yong-feng; REN Nan-qi; YANG Chuan-ping; XU Jing-li

    2006-01-01

    As a new clean energy source, the utilization and demand for hydrogen fuel are rapidly increasing. The integrated process of wastewater treatment of DESAR and energy recovery was developed in the studies. A new hydrogen anaerobe was isolated from the activated sludge. The optimal glucose concentration and the optimal initial pH were 12.0 g/L and 5. 5 respectively. The optimum C/N of the growth and hydrogen production in Rennanqilyf3 was (3.0 ~3.5): 1. The integrated process between DESAR system and biohydrogen production will be an important progress on energy recovery of DESAR system.

  16. The role of acid incubation in rapid immobilization of hydrogen-producing culture in anaerobic upflow column reactors

    Energy Technology Data Exchange (ETDEWEB)

    Zhang, Zhen-Peng; Tay, Joo-Hwa [School of Civil and Environmental Engineering, Nanyang Technological University (Singapore); Institute of Environmental Science and Engineering, Nanyang Technological University (Singapore); Show, Kuan-Yeow [Faculty of Science, Engineering and Technology, University Tunku Abdul Rahman, 31900 Kampar, Perak (Malaysia); Liang, David Tee [Institute of Environmental Science and Engineering, Nanyang Technological University (Singapore); Lee, Duu-Jong [Department of Chemical Engineering, National Taiwan University, Taipei 10617 (China); Su, Ay [Department of Mechanical Engineering, Fuel Cell Center, Yuan-Ze University, Taoyuan 320 (China)

    2008-10-15

    An approach of acidification was examined on formation of hydrogen-producing granules and biofilms in upflow column-shaped reactors. The reactors were fed with synthetic glucose wastewater and operated at 37 C and pH 5.5. The acclimated anaerobic culture was inoculated in four reactors designated R1, R2, R3 and R4, with R3 and R4 filled with granular activated carbon as support medium. To unveil the roles of acidification, microbial culture in R2 and R3 was subject to an acid incubation for 24 h by shifting the culture pH from 5.5 to 2.0. The experimental results suggested that the acidification substantially accelerated microbial granulation, but not biofilm formation. Microbial activities were inhibited by the acid incubation for about 78 h, resulting in the retarded formation of biofilms of the acidified culture. Reducing culture pH resulted in improvement in cell surface physicochemical properties favoring microbial adhesion and immobilization. Zeta potential increased from -25.3 mV to 11.9 mV, hydrophobicity in terms of contact angle improved from 31 to 38 and production of extracellular polymers increased from 66 mg/g-VSS to 136 mg/g-VSS. As a result of the formation of granules and biofilms, high hydrogen production rates of 6.98 and 7.49 L/L h were achieved in granule-based and biofilm-based reactors, respectively. It is concluded that acid incubation is an efficient means to initiate the rapid formation of granules by regulating the surface characteristics of microbial culture. The use of support media as starting nuclei may result in rapid formation of biofilms without the acidification. (author)

  17. 需氧与厌氧配对培养在提高血培养阳性率中的优势%Advantages of aerobic and anaerobic paired culture on raising positive rate of blood culture

    Institute of Scientific and Technical Information of China (English)

    蒋伟燕; 李方去; 杨锦红; 王大选; 刘彩霞

    2011-01-01

    目的 探讨同时做需氧与厌氧配对培养在提高血培养阳性率中的优势.方法 在每例患者的不同部位同时抽取两份血液标本,一份注人需氧血培养瓶,一份注入厌氧血培养瓶,两者分别采用Bact/Alert3D培养仪和BACTEC9120培养仪进行培养.结果 在3605份血培养中阳性共347份,阳性率为9.62%,其中仅需氧血培养阳性120份,占34.58%,仅厌氧血培养阳性88份,占25.36%,需氧与厌氧血培养均阳性139份,占40.06%;在88份仅厌氧血培养阳性中检出厌氧菌3份,占总阳性份数的0.86%.结论 同时做需氧厌氧配对培养能够提高血培养的阳性率,常规开展厌氧血培养十分必要.%OBJECTIVE To discuss the advantage of aerobic and anaerobic paired culture on raising the positive rate of blood culture. METHODS Two samples were collected from each patients different sites at the same time, one was injected into an aerobic blood culture bottle,and the other was injected into an anaerobic blood culture bottle. The aerobic and anaerobic blood culture bottles were cultured by using Baet/Alert 3D cultivator and BACTEC 9120 cultivator respectively. RESULTS There were 347 isolates(9. 62%) recovered from 3605 paired aerobic and anaerobic blood culture bottles, 120 positive isolates (34. 58 %) recovered only from aerobic bottles, and 88 isolates (25. 36 %) recovered only from anaerobic bottles , 139 isolates (40.06 % ) recovered from both of the two bottles. 3 cases of anaerobic bacteria were recovered in 88 isolates, accounting for 0.86%. CONCLUSION Using paired aerobic and anaerobic blood culture bottles can raise the positive rate of blood culture. It is necessary to carry out anaerobic blood culture generally..

  18. Enrichment and characterization of sulfate reducing, naphthalene degrading microorganisms

    Science.gov (United States)

    Steffen, Kümmel; Florian-Alexander, Herbst; Márcia, Duarte; Dietmar, Pieper; Jana, Seifert; Bergen Martin, von; Hans-Hermann, Richnow; Carsten, Vogt

    2014-05-01

    Polycyclic aromatic hydrocarbons (PAH) are pollutants of great concern due to their potential toxicity, mutagenicity and carcinogenicity. PAH are widely distributed in the environment by accidental discharges during the transport, use and disposal of petroleum products, and during forest and grass fires. Caused by their hydrophobic nature, PAH basically accumulate in sediments from where they are slowly released into the groundwater. Although generally limited by the low water solubility of PAH, microbial degradation is one of the major mechanisms leading to the complete clean-up of PAH-contaminated sites. Whereas organisms and biochemical pathways responsible for the aerobic breakdown of PAH are well known, anaerobic PAH biodegradation is less understood; only a few anaerobic PAH degrading cultures have been described. We studied the anaerobic PAH degradation in a microcosm approach to enrich anaerobic PAH degraders. Anoxic groundwater and sediment samples were used as inoculum. Groundwater samples were purchased from the erstwhile gas works facility and a former wood impregnation site. In contrast, sources of sediment samples were a former coal refining area and an old fuel depot. Samples were incubated in anoxic mineral salt medium with naphthalene as sole carbon source and sulfate as terminal electron acceptor. Grown cultures were characterized by feeding with 13C-labeled naphthalene, 16S rRNA gene sequencing using an Illumina® approach, and functional proteome analyses. Finally, six enrichment cultures able to degrade naphthalene under anoxic conditions were established. First results point to a dominance of identified sequences affiliated to the freshwater sulfate-reducing strain N47, which is a known anaerobic naphthalene degrader, in four out of the six enrichments. In those enrichments, peptides related to the pathway of anoxic naphthalene degradation in N47 were abundant. Overall the data underlines the importance of Desulfobacteria for natural

  19. Continuous culture enrichments of ammonia-oxidizing bacteria at low ammonium concentrations

    NARCIS (Netherlands)

    Bollmann, A.; Laanbroek, H.J.

    2001-01-01

    Until now enrichments of ammonia-oxidizing bacteria from natural ammonium-limited environments have been performed mainly in the presence of much higher ammonia concentrations than those present in the natural environment and many have resulted in the enrichment and isolation of environmentally less

  20. Enrichment culture and molecular identification of diverse Bartonella species in stray dogs.

    Science.gov (United States)

    Bai, Ying; Kosoy, Michael Y; Boonmar, Sumalee; Sawatwong, Pongpun; Sangmaneedet, Somboon; Peruski, Leonard F

    2010-12-15

    Using pre-enrichment culture in Bartonella alpha-Proteobacteria growth medium (BAPGM) followed by PCR amplification and DNA sequence identification that targeted a fragment of the citrate synthase gene (gltA), we provide evidence of common bartonella infections and diverse Bartonella species in the blood of stray dogs from Bangkok and Khon Kaen, Thailand. The overall prevalence of all Bartonella species was 31.3% (60/192), with 27.9% (31/111) and 35.8% (29/81) in the stray dogs from Bangkok and Khon Kaen, respectively. Phylogenetic analyzes of gltA identified eight species/genotypes of Bartonella in the blood of stray dogs, including B. vinsonii subsp. arupensis, B. elizabethae, B. grahamii, B. quintana, B. taylorii, and three novel genotypes (BK1, KK1 and KK2) possibly representing unique species with ≤ 90.2% similarities to any of the known Bartonella species B. vinsonii subsp. arupensis was the only species detected in dogs from both sites, B. quintana and BK1 were found in the dogs from Bangkok, B. elizabethae, B. taylorii, KK1 and KK2 were found in the dogs from Khon Kaen. We conclude that stray dogs in Thailand are frequently infected with Bartonella species that vary with geographic region. As some Bartonella species detected in the present study are considered pathogenic for humans, stray dogs in Thailand may serve as possible reservoirs for Bartonella causing human illnesses. Further work is needed to determine the role of those newly discovered Bartonella genotypes/species in human and veterinary medicine.

  1. Bacterial community analysis in chlorpyrifos enrichment cultures via DGGE and use of bacterial consortium for CP biodegradation.

    Science.gov (United States)

    Akbar, Shamsa; Sultan, Sikander; Kertesz, Michael

    2014-10-01

    The organophosphate pesticide chlorpyrifos (CP) has been used extensively since the 1960s for insect control. However, its toxic effects on mammals and persistence in environment necessitate its removal from contaminated sites, biodegradation studies of CP-degrading microbes are therefore of immense importance. Samples from a Pakistani agricultural soil with an extensive history of CP application were used to prepare enrichment cultures using CP as sole carbon source for bacterial community analysis and isolation of CP metabolizing bacteria. Bacterial community analysis (denaturing gradient gel electrophoresis) revealed that the dominant genera enriched under these conditions were Pseudomonas, Acinetobacter and Stenotrophomonas, along with lower numbers of Sphingomonas, Agrobacterium and Burkholderia. Furthermore, it revealed that members of Bacteroidetes, Firmicutes, α- and γ-Proteobacteria and Actinobacteria were present at initial steps of enrichment whereas β-Proteobacteria appeared in later steps and only Proteobacteria were selected by enrichment culturing. However, when CP-degrading strains were isolated from this enrichment culture, the most active organisms were strains of Acinetobacter calcoaceticus, Pseudomonas mendocina and Pseudomonas aeruginosa. These strains degraded 6-7.4 mg L(-1) day(-1) of CP when cultivated in mineral medium, while the consortium of all four strains degraded 9.2 mg L(-1) day(-1) of CP (100 mg L(-1)). Addition of glucose as an additional C source increased the degradation capacity by 8-14 %. After inoculation of contaminated soil with CP (200 mg kg(-1)) disappearance rates were 3.83-4.30 mg kg(-1) day(-1) for individual strains and 4.76 mg kg(-1) day(-1) for the consortium. These results indicate that these organisms are involved in the degradation of CP in soil and represent valuable candidates for in situ bioremediation of contaminated soils and waters.

  2. Assessing the influence of CH4 concentration during culture enrichment on the biodegradation kinetics and population structure.

    Science.gov (United States)

    López, Juan C; Quijano, Guillermo; Pérez, Rebeca; Muñoz, Raúl

    2014-12-15

    Methanotrophic communities were enriched in three stirred tank reactors continuously supplied with CH4-laden air at 20, 2 and 0.2 gCH4 m(-3) in order to evaluate the influence of CH4 concentration on the biodegradation kinetics, population structure and potential polyhydroxyalkanoate production under sequential nitrogen limitations. The population structure of the enriched cultures, dominated by type I methanotrophs, was influenced by CH4 concentration. No significant correlation between CH4 concentration and the maximum specific degradation rate (qmax) or the half-saturation constant (KS) was recorded, microorganisms enriched at 2 gCH4 m(-3) presenting the highest qmax and those enriched at 20 and 0.2 gCH4 m(-3) exhibiting the lowest KS. Maximum polyhydroxybutyrate (PHB) contents of 1.0% and 12.6% (w/w) were achieved at 20 and 2 g CH4 m(-3), respectively. Polyhydroxyvalerate (PHV) was also detected at PHV:PHB ratios of up to 12:1 and 4:1 in the communities enriched at 20 and 0.2 gCH4 m(-3), respectively.

  3. Anaerobic taurine oxidation: a novel reaction by a nitrate-reducing Alcaligenes sp.

    Science.gov (United States)

    Denger, K; Laue, H; Cook, A M

    1997-06-01

    Enrichment cultures were prepared under strictly anoxic conditions in medium representing fresh water and containing an organosulfonate as electron donor and carbon source, and nitrate as electron acceptor. The inoculum was from the anaerobic digestor of two communal sewage works. The natural organosulfonates 2-aminoethanesulfonate (taurine), DL-2-amino-3-sulfopropionate (cysteate) and 2-hydroxyethanesulfonate (isethionate) all gave positive enrichments, whereas unsubstituted alkanesulfonates, such as methanesulfonate and arenesulfonates, gave no enrichment. Two representative enrichments were used to obtain pure cultures, and strains NKNTAU (utilizing taurine) and NKNIS (utilizing isethionate) were isolated. Strain NKNTAU was examined in detail. Out of 18 tested organosulfonates, it utilized only one, taurine, and was identified as a novel Alcaligenes sp., a facultatively anaerobic bacterium. Carbon from taurine was converted to cell material and carbon dioxide. The amino group was released as ammonium ion and the sulfonate moiety was recovered as sulfate. Nitrate was reduced to nitrogen gas.

  4. Potential effect of matrix stiffness on the enrichment of tumor initiating cells under three-dimensional culture conditions.

    Science.gov (United States)

    Liu, Chang; Liu, Yang; Xu, Xiao-xi; Wu, Hao; Xie, Hong-guo; Chen, Li; Lu, Ting; Yang, Li; Guo, Xin; Sun, Guang-wei; Wang, Wei; Ma, Xiao-jun; He, Xin

    2015-01-01

    Cancer stem cell (CSC) or tumor initiating cell (TIC) plays an important role in tumor progression and metastasis. Biophysical forces in tumor microenvironment have an important effect on tumor formation and development. In this study, the potential effect of matrix stiffness on the biological characteristics of human head and neck squamous cell carcinoma (HNSCC) TICs, especially the enrichment of HNSCC TICs, was investigated under three-dimensional (3D) culture conditions by means of alginate gel (ALG) beads with different matrix stiffnesses. ALG beads with soft (21 kPa), moderate (70 kPa) and hard (105 kPa) stiffness were generated by changing alginate concentration. It was found that significant HNSCC TIC enrichment was achieved in the ALG beads with moderate matrix stiffness (70 kPa). The gene expression of stemness markers Oct3/4 and Nanog, TIC markers CD44 and ABCG2 was enhanced in cells under this moderate (70 kPa) stiffness. HNSCC TIC proportion was also highly enriched under moderate matrix stiffness, accompanying with higher tumorigenicity, metastatic ability and drug resistance. And it was also found that the possible molecular mechanism underlying the regulated TIC properties by matrix stiffness under 3D culture conditions was significantly different from 2D culture condition. Therefore, the results achieved in this study indicated that 3D biophysical microenvironment had an important effect on TIC characteristics and alginate-based biomimetic scaffolds could be utilized as a proper platform to investigate the interaction between tumor cells and 3D microenvironment.

  5. Building Learning Communities for Research Collaboration and Cross-Cultural Enrichment in Science Education

    Science.gov (United States)

    Sparrow, E. B.

    2003-12-01

    The GLOBE program has provided opportunities for environmental science research and education collaborations among scientists, teachers and K-12 students, and for cross-cultural enrichment nationally and abroad. In Alaska, GLOBE has also provided funding leverage in some cases, and a base for several other science education programs that share a common goal of increasing student interest, understanding, process skills and achievement in science, through involvement in ongoing research investigations. These programs that use GLOBE methodologies (standardized scientific measurements and learning activities developed by scientists and educators) are: Global Change Education Using Western Science and Native Knowledge also known as "Observing Locally, Connecting Globally" (OLCG); Alaska Earth System Science Education Alliance: Improving Understanding of Climate Variability and Its Relevance to Rural Alaska; Schoolyard Long Term Ecological Research; Alaska Rural Research Partnership; Alaska Partnership for Teacher Enhancement; Alaska Lake Ice and Snow Observatory Network; Alaska Boreal Forest Council Education Outreach; Calypso Farm and Ecology Center; Environmental Education Outreach; and also GLOBE Arctic POPs (persistent organic pollutants) a program that involves countries in the circumpolar North. The University of Alaska GLOBE Partnership has collaborated with the BLM Campbell Creek Science Center Globe Partnership in facilitating GLOBE Training Workshops and providing teacher support. GLOBE's extensive website including data entry, archive, analysis and visualization capabilities; GLOBE Teacher Guide, videos and other materials provided; excellent GLOBE science research and education staff, training support office, GLOBE help desk, alignment of GLOBE curriculum with national science education standards and GLOBE certification of teachers trained on even just one GLOBE investigation, have made it easier to implement GLOBE in the classroom. Using GLOBE, whole

  6. Evidence for the biogenic origin of manganese-enriched layers in Lake Superior sediments.

    Science.gov (United States)

    Palermo, Christine; Dittrich, Maria

    2016-04-01

    Manganese (Mn) and iron (Fe)-enriched sediment layers were discovered in Lake Superior within, above and below the oxic-anoxic interface. While the role of bacteria in redox reactions with Mn is known to be significant, little information exists about indigenous microbial communities in many freshwater environments. This study examined the bacterial communities of Mn-enriched layers in Lake Superior to identify the potential Mn(II) oxidizers responsible for the formation of Mn oxides. Anaerobic Mn(II) oxidation occurring in the Mn-enriched layers at the oxic-anoxic interface was investigated using Mn(II)-enriched cultures. High-resolution microscopic and spectroscopic investigations provided evidence of the biogenic formation of Mn oxides on cell surfaces. Spectroscopic mapping confirmed high levels of Mn in structures resembling biogenic Mn oxides. These structures were observed in enrichment cultures and in Mn-enriched layer sediment samples, indicating the significance of biogenic Mn oxidation occurring in situ. 16S ribosomal DNA pyrosequencing was used to identify the bacteria potentially responsible for Mnoxide formation in the enrichment cultures and Mn-enriched layers, revealing that the Mn-enriched layer contains classes with known Mn(II)-oxidizing members. Pyrosequencing of bacterial cultures suggested that these bacteria may be Bacillus strains, and that anaerobic microbial-mediated Mn(II) oxidation contributes to the formation of the layers.

  7. Anaerobic bacteria in otitis media.

    Science.gov (United States)

    Fulghum, R S; Daniel, H J; Yarborough, J G

    1977-01-01

    Anaerobic bacteria, Peptostrepotococcus intermedius and Propionibacterium acnes, were found in mixed culture specimens from four to ten tested cases of chronic secretory otitis media. These anaerobic bacteria were in a mixed infection flora with aerobic bacteria most often Staphylococcus epidermidis and Cornybacterium sp. which do not fit any established species. The findings of anaerobic bacteria in otitis media is consistent with the sporadic report of the involvement of anaerobic bacteria in otitis media in the literature since 1898.

  8. Diversity of anaerobic microbes in spacecraft assembly clean rooms.

    Science.gov (United States)

    Probst, Alexander; Vaishampayan, Parag; Osman, Shariff; Moissl-Eichinger, Christine; Andersen, Gary L; Venkateswaran, Kasthuri

    2010-05-01

    Although the cultivable and noncultivable microbial diversity of spacecraft assembly clean rooms has been previously documented using conventional and state-of-the-art molecular techniques, the occurrence of obligate anaerobes within these clean rooms is still uncertain. Therefore, anaerobic bacterial communities of three clean-room facilities were analyzed during assembly of the Mars Science Laboratory rover. Anaerobic bacteria were cultured on several media, and DNA was extracted from suitable anaerobic enrichments and examined with conventional 16S rRNA gene clone library, as well as high-density phylogenetic 16S rRNA gene microarray (PhyloChip) technologies. The culture-dependent analyses predominantly showed the presence of clostridial and propionibacterial strains. The 16S rRNA gene sequences retrieved from clone libraries revealed distinct microbial populations associated with each clean-room facility, clustered exclusively within gram-positive organisms. PhyloChip analysis detected a greater microbial diversity, spanning many phyla of bacteria, and provided a deeper insight into the microbial community structure of the clean-room facilities. This study presents an integrated approach for assessing the anaerobic microbial population within clean-room facilities, using both molecular and cultivation-based analyses. The results reveal that highly diverse anaerobic bacterial populations persist in the clean rooms even after the imposition of rigorous maintenance programs and will pose a challenge to planetary protection implementation activities.

  9. The culture of cancer cell lines as tumorspheres does not systematically result in cancer stem cell enrichment.

    Directory of Open Access Journals (Sweden)

    Christophe Y Calvet

    Full Text Available Cancer stem cells (CSC have raised great excitement during the last decade and are promising targets for an efficient treatment of tumors without relapses and metastases. Among the various methods that enable to enrich cancer cell lines in CSC, tumorspheres culture has been predominantly used. In this report, we attempted to generate tumorspheres from several murine and human cancer cell lines: B16-F10, HT-29, MCF-7 and MDA-MB-231 cells. Tumorspheres were obtained with variable efficiencies from all cell lines except from MDA-MB-231 cells. Then, we studied several CSC characteristics in both tumorspheres and adherent cultures of the B16-F10, HT-29 and MCF-7 cells. Unexpectedly, tumorspheres-forming cells were less clonogenic and, in the case of B16-F10, less proliferative than attached cells. In addition, we did not observe any enrichment in the population expressing CSC surface markers in tumorspheres from B16-F10 (CD133, CD44 and CD24 markers or MCF-7 (CD44 and CD24 markers cells. On the contrary, tumorspheres culture of HT-29 cells appeared to enrich in cells expressing colon CSC markers, i.e. CD133 and CD44 proteins. For the B16-F10 cell line, when 1 000 cells were injected in syngenic C57BL/6 mice, tumorspheres-forming cells displayed a significantly lower tumorigenic potential than adherent cells. Finally, tumorspheres culture of B16-F10 cells induced a down-regulation of vimentin which could explain, at least partially, the lower tumorigenicity of tumorspheres-forming cells. All these results, along with the literature, indicate that tumorspheres culture of cancer cell lines can induce an enrichment in CSC but in a cell line-dependent manner. In conclusion, extensive characterization of CSC properties in tumorspheres derived from any cancer cell line or cancer tissue must be performed in order to ensure that the generated tumorspheres are actually enriched in CSC.

  10. Effect of free ammonia and free nitrous acid concentration on the anabolic and catabolic processes of an enriched Nitrosomonas culture.

    Science.gov (United States)

    Vadivelu, Vel M; Keller, Jurg; Yuan, Zhiguo

    2006-12-05

    The effects of free ammonia (FA; NH(3)) and free nitrous acid (FNA; HNO(2)) concentrations on the metabolisms of an enriched ammonia oxidizing bacteria (AOB) culture were investigated using a method allowing the decoupling of growth and energy generation processes. A lab-scale sequencing batch reactor (SBR) was operated for the enrichment of an AOB culture. Fluorescent in-situ hybridization (FISH) analysis showed that 82% of the bacterial population in the SBR bound to the NEU probe specifically designed for Nitrosomonas europaea. Batch tests were carried out to measure the oxygen and ammonium consumption rates by the culture at various FA and FNA levels, in the presence or absence of inorganic carbon (CO(2), HCO(3) (-), and CO(3) (2-)). It was revealed that FA of up to 16.0 mgNH(3)-N . L(-1), which was the highest concentration used in this study, did not have any inhibitory effect on either the catabolic or anabolic processes of the Nitrosomonas culture. In contrast, FNA inhibited both the growth and energy production capabilities of the Nitrosomonas culture. The inhibition on growth initiated at approximately 0.10 mgHNO(2)-N . L(-1), and the data suggested that the biosynthesis was completely stopped at an FNA concentration of 0.40 mgHNO(2)-N . L(-1). The inhibition on energy generation initiated at a slightly lower level but the Nitrosomonas culture was still oxidizing ammonia at half of the maximum rate at an FNA concentration of 0.50-0.63 mgHNO(2)-N . L(-1). The affinity constant of the Nitrosomonas culture with respect to ammonia was determined to be 0.36 mgNH(3)-N . L(-1), independent of the presence or absence of inorganic carbon.

  11. Effect of changing temperature on anaerobic hydrogen production and microbial community composition in an open-mixed culture bioreactor

    Energy Technology Data Exchange (ETDEWEB)

    Karadag, Dogan; Puhakka, Jaakko A. [Department of Chemistry and Bioengineering, Tampere University of Technology, Tampere (Finland)

    2010-10-15

    The temperature effect (37-65 C) on H{sub 2} production from glucose in an open-mixed culture bioreactor using an enrichment culture from a hot spring was studied. The dynamics of microbial communities was investigated by polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE). At 45 and 60 C the H{sub 2} production was the highest i.e. 1.71 and 0.85 mol H{sub 2}/mol glucose, respectively. No H{sub 2} was produced at temperatures 50 and 55 C. At 37-45 C, H{sub 2} production was produced by butyrate type fermentation while fermentation mechanism changed to ethanol type at 60 C. Clostridium species were dominant at 37-45 C while at 50-55 C and 60 C the culture was dominated by Bacillus coagulans and Thermoanaerobacterium, respectively. In the presence of B. Coagulans the metabolism was directed to lactate production. The results show that the mixed culture had two optima for H{sub 2} production and that the microbial communities and metabolic patterns promptly changed according to changing temperatures. (author)

  12. Biodegradation of azo dyes in cocultures of anaerobic granular sludge with aerobic aromatic amine degrading enrichment cultures

    NARCIS (Netherlands)

    Tan, N.C.G.; Prenefeta-Boldú, F.X.; Opsteeg, J.L.; Lettinga, G.; Field, J.A.

    1999-01-01

    A prerequisite for the mineralization (complete biodegradation) of many azo dyes is a combination of reductive and oxidative steps. In this study, the biodegradation of two azo dyes, 4-phenylazophenol (4-PAP) and Mordant Yellow 10 (4-sulfophenylazo-salicylic acid; MY10), was evaluated in batch exper

  13. Improved enrichment culture technique for methane-oxidizing bacteria from marine ecosystems: the effect of adhesion material and gas composition.

    Science.gov (United States)

    Vekeman, Bram; Dumolin, Charles; De Vos, Paul; Heylen, Kim

    2017-02-01

    Cultivation of microbial representatives of specific functional guilds from environmental samples depends largely on the suitability of the applied growth conditions. Especially the cultivation of marine methanotrophs has received little attention, resulting in only a limited number of ex situ cultures available. In this study we investigated the effect of adhesion material and headspace composition on the methane oxidation activity in methanotrophic enrichments obtained from marine sediment. Addition of sterilized natural sediment or alternatively the addition of acid-washed silicon dioxide significantly increased methane oxidation. This positive effect was attributed to bacterial adhesion on the particles via extracellular compounds, with a minimum amount of particles required for effect. As a result, the particles were immobilized, thus creating a stratified environment in which a limited diffusive gas gradients could build up and various microniches were formed. Such diffusive gas gradient might necessitate high headspace concentrations of CH4 and CO2 for sufficient concentrations to reach the methane-oxidizing bacteria in the enrichment culture technique. Therefore, high concentrations of methane and carbon dioxide, in addition to the addition of adhesion material, were tested and indeed further stimulated methane oxidation. Use of adhesion material in combination with high concentrations of methane and carbon dioxide might thus facilitate the cultivation and subsequent enrichment of environmentally important members of this functional guild. The exact mechanism of the observed positive effects on methane oxidation and the differential effect on methanotrophic diversity still needs to be explored.

  14. Enrichment strategy to select functional consortium from mixed cultures: Consortium from rumen liquor for simultaneous cellulose degradation and hydrogen production

    Energy Technology Data Exchange (ETDEWEB)

    Wang, Aijie; Ren, Nanqi [State Key Lab of Urban Water Resource and Environment, Harbin Institute of Technology, Harbin 150090 (China); School of Environmental and Municipal Engineering, Harbin Institute of Technology, Harbin 150090 (China); Gao, Lingfang; Xu, Jifei; Liu, Chong; Lee, Duu-Jong [School of Environmental and Municipal Engineering, Harbin Institute of Technology, Harbin 150090 (China)

    2010-12-15

    Strain isolation using conventional roll tube/plating technique is time consuming and is able to culture in vitro only a small fraction of existing microbes in a natural microflora. This paper proposed a simple and rapid method to select the as-simple-as-possible biological consortium by serially diluting the original mixed culture. The diluted which remains, while the one diluted in serial loses the target function, is defined as the functional consortium of the original mixed culture. Since the microbial structure and the reaction pathway incorporated with the functional consortium is much simpler than its original mother liquor, detailed analysis on the strain interaction is possible without the risk of losing key functional strains as often caused from conventional isolation method. The rumen liquor that can degrade cellulose and produce hydrogen is used as a demonstration example. A ''rumen cellulose-degrading bacterial consortium'' (RCBC) was identified using the proposed enrichment strategy. (author)

  15. Proteomic characterization of golgi membranes enriched from Arabidopsis suspension cell cultures

    DEFF Research Database (Denmark)

    Hansen, Sara Fasmer; Ebert, Berit; Rautengarten, Carsten

    2016-01-01

    The plant Golgi apparatus has a central role in the secretory pathway and is the principal site within the cell for the assembly and processing of macromolecules. The stacked membrane structure of the Golgi apparatus along with its interactions with the cytoskeleton and endoplasmic reticulum has...... historically made the isolation and purification of this organelle difficult. Density centrifugation has typically been used to enrich Golgi membranes from plant microsomal preparations, and aside from minor adaptations, the approach is still widely employed. Here we outline the enrichment of Golgi membranes...

  16. Metaproteogenomic analysis of a sulfate-reducing enrichment culture reveals genomic organization of key enzymes in the m-xylene degradation pathway and metabolic activity of proteobacteria.

    Science.gov (United States)

    Bozinovski, Dragana; Taubert, Martin; Kleinsteuber, Sabine; Richnow, Hans-Hermann; von Bergen, Martin; Vogt, Carsten; Seifert, Jana

    2014-10-01

    This study aimed to ascertain the functional and phylogenetic relationships within an m-xylene degrading sulfate-reducing enrichment culture, which had been maintained for several years in the laboratory with m-xylene as the sole source of carbon and energy. Previous studies indicated that a phylotype affiliated to the Desulfobacteraceae was the main m-xylene assimilating organism. In the present study, genes and gene products were identified by a metaproteogenomic approach using LC-MS/MS analysis of the microbial community, and 2426 peptides were identified from 576 proteins. In the metagenome of the community, gene clusters encoding enzymes involved in fumarate addition to a methyl moiety of m-xylene (nms, bss), as well as gene clusters coding for enzymes involved in modified beta-oxidation to (3-methyl)benzoyl-CoA (bns), were identified in two separate contigs. Additionally, gene clusters containing homologues to bam genes encoding benzoyl-CoA reductase (Bcr) class II, catalyzing the dearomatization of (3-methyl)benzoyl-CoA, were identified. Time-resolved protein stable isotope probing (protein-SIP) experiments using (13)C-labeled m-xylene showed that the respective gene products were highly (13)C-labeled. The present data suggested the identification of gene products that were similar to those involved in methylnaphthalene degradation even though the consortium was not capable of growing in the presence of naphthalene, methylnaphthalene or toluene as substrates. Thus, a novel branch of enzymes was found that was probably specific for anaerobic m-xylene degradation.

  17. A novel anaerobic co-culture system for bio-hydrogen production from sugarcane bagasse.

    Science.gov (United States)

    Cheng, Jingrong; Zhu, Mingjun

    2013-09-01

    A novel co-culture of Clostridium thermocellum and Thermoanaerobacterium aotearoense with pretreated sugarcane bagasse (SCB) under mild alkali conditions for bio-hydrogen production was established, exhibiting a cost-effective and synergetic advantage in bio-hydrogen production over monoculture of C. thermocellum or T. aotearoense with untreated SCB. The optimized pretreatment conditions were established to be 3% NaOH, and a liquid to solid ratio of 25:1 at 80°C for 3h. A final hydrogen production of 50.05±1.51 mmol/L was achieved with 40 g/L pretreated SCB at 55°C. The established co-culture system provides a novel consolidated bio-processing strategy for bioconversion of SCB to bio-hydrogen.

  18. Biosynthesis of highly enriched 13C-lycopene for human metabolic studies using repeated batch tomato cell culturing with 13C-glucose

    OpenAIRE

    Moran, Nancy E.; Rogers, Randy B.; Lu, Chi-Hua; Conlon, Lauren E.; Lila, Mary Ann; Clinton, Steven K.; Erdman, John W

    2013-01-01

    While putative disease-preventing lycopene metabolites are found in both tomato (Solanum lycopersicum) products and in their consumers, mammalian lycopene metabolism is poorly understood. Advances in tomato cell culturing techniques offer an economical tool for generation of highly-enriched 13C-lycopene for human bioavailability and metabolism studies. To enhance the 13C-enrichment and yields of labeled lycopene from the hp-1 tomato cell line, cultures were first grown in 13C-glucose media fo...

  19. Effects of carbon sources on the enrichment of halophilic polyhydroxyalkanoate-storing mixed microbial culture in an aerobic dynamic feeding process

    Science.gov (United States)

    Cui, You-Wei; Zhang, Hong-Yu; Lu, Peng-Fei; Peng, Yong-Zhen

    2016-08-01

    Microbial polyhydroxyalkanoate (PHA) production serves as a substitute for petroleum-based plastics. Enriching mixed microbial cultures (MMCs) with the capacity to store PHA is a key precursor for low-cost PHA production. This study investigated the impact of carbon types on enrichment outcomes. Three MMCs were separately fed by acetate sodium, glucose, and starch as an enriching carbon source, and were exposed to long-term aerobic dynamic feeding (ADF) periods. The PHA production capacity, kinetics and stoichiometry of the enrichments, the PHA composition, and the microbial diversity and community composition were explored to determine carbon and enrichment correlations. After 350-cycle enriching periods under feast-famine (F-F) regimes, the MMCs enriched by acetate sodium and glucose contained a maximum PHA content of 64.7% and 60.5% cell dry weight (CDW). The starch-enriched MMC only had 27.3% CDW of PHA. High-throughput sequencing revealed that non-PHA bacteria survived alongside PHA storing bacteria, even under severe F-F selective pressure. Genus of Pseudomonas and Stappia were the possible PHA accumulating bacteria in acetate-enriched MMC. Genus of Oceanicella, Piscicoccus and Vibrio were found as PHA accumulating bacteria in glucose-enriched MMC. Vibrio genus was the only PHA accumulating bacteria in starch-enriched MMC. The community diversity and composition were regulated by the substrate types.

  20. Thermo-acidophillic biohydrogen production from rice bran de-oiled wastewater by Selectively enriched mixed culture

    Energy Technology Data Exchange (ETDEWEB)

    Sivaramakrishna, D.; Sreekanth, D.; Himabindu, V. [Centre for Environment, Institute of Science and Technology, Jawaharlal Nehru Technological University Hyderabad, Kukatpally Hyderabad-500 085 (India); Narasu, M. Lakshmi [Centre for Biotechnology, Institute of Science and Technology, Jawaharlal Nehru Technological University Hyderabad, Kukatpally Hyderabad-500 085 (India)

    2010-07-01

    The present study focuses on the biohydrogen production in an anaerobic batch reactor operated at thermophillic (570C) and acidophilic conditions (pH 6) with rice bran de-oiled wastewater (RBOW) as substrate. The hydrogen generating mixed microflora was enriched from slaughter house sludge (SHS) through acid treatment (pH 3-4, for 24h) coupled with heat treatment (1h at 1000C) to eliminate non-spore forming bacteria and to inhibit the growth of methanogenic bacteria (MB) prior to inoculation in the reactor. The hydrogen production rate was maximum at 570C (1861 +- 14ml/L-WW/d) compared to 370C (651 +- 30ml/L-ww/d). The Hydrogen yield increased with temperature from 1.1 to 2.2 molH2/mol of substrate respectively. The optimum pH range for hydrogen production in this system was observed in between 5.5 to 6. Acid-forming pathway with butyric acid as a major metabolite dominated the metabolic flow during the hydrogen production.

  1. Thermo-acidophillic biohydrogen production from rice bran de-oiled wastewater by Selectively enriched mixed culture

    Directory of Open Access Journals (Sweden)

    D.Sivaramakrishna, D.Sreekanth, V.Himabindu, M.Lakshmi Narasu

    2010-07-01

    Full Text Available The present study focuses on the biohydrogen production in an anaerobic batch reactor operated at thermophillic (570C and acidophilic conditions (pH 6 with rice bran de-oiled wastewater (RBOW as substrate. The hydrogen generating mixed microflora was enriched from slaughter house sludge (SHS through acid treatment (pH 3-4, for 24h coupled with heat treatment (1h at 1000C to eliminate non-spore forming bacteria and to inhibit the growth of methanogenic bacteria (MB prior to inoculation in the reactor. The hydrogen production rate was maximum at 570C (1861±14ml/L-WW/d compared to 370C (651±30ml/L-ww/d. The Hydrogen yield increased with temperature from 1.1 to 2.2 molH2/mol of substrate respectively. The optimum pH range for hydrogen production in this system was observed in between 5.5 to 6. Acid-forming pathway with butyric acid as a major metabolite dominated the metabolic flow during the hydrogen production.

  2. RISK FACTORS IN NEONATAL ANAEROBIC INFECTIONS

    Directory of Open Access Journals (Sweden)

    M. S. Tabib

    2008-06-01

    Full Text Available Anaerobic bacteria are well known causes of sepsis in adults but there are few studies regarding their role in neonatal sepsis. In an attempt to define the incidence of neonatal anaerobic infections a prospective study was performed during one year period. A total number of 400 neonates under sepsis study were entered this investigation. Anaerobic as well as aerobic cultures were sent. The patients were subjected to comparison in two groups: anaerobic culture positive and anaerobic culture negative and this comparison were analyzed statistically. There were 7 neonates with positive anaerobic culture and 35 neonates with positive aerobic culture. A significant statistical relationship was found between anaerobic infections and abdominal distention and pneumonia. It is recommended for those neonates with abdominal distention and pneumonia refractory to antibiotic treatment to be started on antibiotics with anaerobic coverage.

  3. Metabolic energy-based modelling explains product yielding in anaerobic mixed culture fermentations.

    Directory of Open Access Journals (Sweden)

    Rebeca González-Cabaleiro

    Full Text Available The fermentation of glucose using microbial mixed cultures is of great interest given its potential to convert wastes into valuable products at low cost, however, the difficulties associated with the control of the process still pose important challenges for its industrial implementation. A deeper understanding of the fermentation process involving metabolic and biochemical principles is very necessary to overcome these difficulties. In this work a novel metabolic energy based model is presented that accurately predicts for the first time the experimentally observed changes in product spectrum with pH. The model predicts the observed shift towards formate production at high pH, accompanied with ethanol and acetate production. Acetate (accompanied with a more reduced product and butyrate are predicted main products at low pH. The production of propionate between pH 6 and 8 is also predicted. These results are mechanistically explained for the first time considering the impact that variable proton motive potential and active transport energy costs have in terms of energy harvest over different products yielding. The model results, in line with numerous reported experiments, validate the mechanistic and bioenergetics hypotheses that fermentative mixed cultures products yielding appears to be controlled by the principle of maximum energy harvest and the necessity of balancing the redox equivalents in absence of external electron acceptors.

  4. Anaerobic oxidation of methane in grassland soils used for cattle husbandry

    Science.gov (United States)

    Bannert, A.; Bogen, C.; Esperschütz, J.; Koubová, A.; Buegger, F.; Fischer, D.; Radl, V.; Fuß, R.; Chroňáková, A.; Elhottová, D.; Šimek, M.; Schloter, M.

    2012-10-01

    While the importance of anaerobic methane oxidation has been reported for marine ecosystems, the role of this process in soils is still questionable. Grasslands used as pastures for cattle overwintering show an increase in anaerobic soil micro-sites caused by animal treading and excrement deposition. Therefore, anaerobic potential methane oxidation activity of severely impacted soil from a cattle winter pasture was investigated in an incubation experiment under anaerobic conditions using 13C-labelled methane. We were able to detect a high microbial activity utilizing CH4 as nutrient source shown by the respiration of 13CO2. Measurements of possible terminal electron acceptors for anaerobic oxidation of methane were carried out. Soil sulfate concentrations were too low to explain the oxidation of the amount of methane added, but enough nitrate and iron(III) were detected. However, only nitrate was consumed during the experiment. 13C-PLFA analyses clearly showed the utilization of CH4 as nutrient source mainly by organisms harbouring 16:1ω7 PLFAs. These lipids were also found as most 13C-enriched fatty acids by Raghoebarsing et al. (2006) after addition of 13CH4 to an enrichment culture coupling denitrification of nitrate to anaerobic oxidation of methane. This might be an indication for anaerobic oxidation of methane by relatives of "Candidatus Methylomirabilis oxyfera" in the investigated grassland soil under the conditions of the incubation experiment.

  5. Effects of CO2 enrichment and nutrients supply intermittency on batch cultures of Isochrysis galbana.

    Science.gov (United States)

    Picardo, Marta C; de Medeiros, José Luiz; Araújo, Ofélia de Queiroz F; Chaloub, Ricardo Moreira

    2013-09-01

    Aiming at enhanced performance to increase economic feasibility of microalgae based processes, Isochrysis galbana was grown in three modes of cultivation: batch, intermittent fed batch and semi-continuous. The batch mode was conducted under two regimes of aeration: conventional aeration and CO2 enriched aeration (5% v/v in air). Increased biomass productivity without significant impact on lipid accumulation was observed for CO2 enriched aeration relatively to cultivation aerated with air only. The intermittent fed batch cultivation policy was proven to be useful for lipid accumulation, increasing the lipid content by 19.8%. However, the semi-continuous mode resulted in higher productivity due to increased biomass concentration; the biomass productivity reached 0.51 g/(Ld). Fluorescence measurements were performed; the calculated low electron transport rate showed the need to increase the irradiance. The results showed that I. galbana can be grown in semi-continuous condition at high levels of biomass productivity.

  6. Microbial succession in response to pollutants in batch-enrichment culture

    OpenAIRE

    Shuo Jiao; Weimin Chen; Entao Wang; Junman Wang; Zhenshan Liu; Yining Li; Gehong Wei

    2016-01-01

    As a global problem, environmental pollution is an important factor to shape the microbial communities. The elucidation of the succession of microbial communities in response to pollutants is essential for developing bioremediation procedures. In the present study, ten batches of soil-enrichment subcultures were subjected to four treatments: phenanthrene, n-octadecane, phenanthrene + n-octadecane, or phenanthrene + n-octadecane + CdCl2. Forty pollutant-degrading consortia, corresponding to ea...

  7. Microbial succession in response to pollutants in batch-enrichment culture.

    Science.gov (United States)

    Jiao, Shuo; Chen, Weimin; Wang, Entao; Wang, Junman; Liu, Zhenshan; Li, Yining; Wei, Gehong

    2016-02-24

    As a global problem, environmental pollution is an important factor to shape the microbial communities. The elucidation of the succession of microbial communities in response to pollutants is essential for developing bioremediation procedures. In the present study, ten batches of soil-enrichment subcultures were subjected to four treatments: phenanthrene, n-octadecane, phenanthrene + n-octadecane, or phenanthrene + n-octadecane + CdCl2. Forty pollutant-degrading consortia, corresponding to each batch of the four treatments were obtained. High-throughput sequencing of the 16S rRNA gene revealed that the diversity, richness and evenness of the consortia decreased throughout the subculturing procedure. The well-known hydrocarbon degraders Acinetobacter, Gordonia, Sphingobium, Sphingopyxis, and Castellaniella and several other genera, including Niabella and Naxibacter, were detected in the enriched consortia. The predominant microbes varied and the microbial community in the consortia gradually changed during the successive subculturing depending on the treatment, indicating that the pollutants influenced the microbial successions. Comparison of the networks in the treatments indicated that organic pollutants and CdCl2 affected the co-occurrence patterns in enriched consortia. In conclusion, single environmental factors, such as the addition of nutrients or selection pressure, can shape microbial communities and partially explain the extensive differences in microbial community structures among diverse environments.

  8. Passport to Cultural Enrichment: The Peace Corps World Wise Schools Experience

    Science.gov (United States)

    Carano, Kenneth T.

    2009-01-01

    In recent studies, youths in the United States have demonstrated a remarkable lack of cultural literacy. As the world is becoming increasingly interconnected, it is imperative that students enhance their understanding of other cultures. A classroom correspondence match with a Peace Corps volunteer through the Coverdell Peace Corps World Wise…

  9. Signatures of Autotrophic and Heterotrophic Metabolic Activity in Enrichment Cultures from a Sulphur Oxidizing Acid Mine Site

    Science.gov (United States)

    Slater, G. F.; Bernier, L.; Cowie, B. R.; Warren, L. A.

    2006-12-01

    Delineating the role of microorganisms in geochemical processes of interest in natural environments requires the development of tools that provide the ability to distinguish amongst microbial activity associated with different metabolic guilds. The gap between phylogenetic characterization and phenotypic understanding remains, underscoring the need to consider alternative methods. Compound specific analysis of cellular components has the potential to differentiate between active metabolic processes supporting microbial communities and may be especially useful in extreme environments. The goal of this study was to determine whether the phospholipids fatty acid (PLFA) distribution and isotopic signatures associated with autotrophs and heterotrophs enriched from an acid mine drainage (AMD) system differed, and further whether natural consortial autotrophic isolates showed similar signatures to autotrophic pure strains of Acidithiobacillus ferrooxidans and A. thiooxidans. Two distinct initial enrichments with tetrathionate and CO2 yielded primarily autotrophic (95%) Acidithiobaccillus spp. sulphur oxidizing communities. The remaining microbial members of theses enrichments (subculture of the consortial isolates in a medium amended with glucose but without tetrathionate selectively resulted in their visible growth. PLFA profiles and δ13C signatures from autotrophic (1) natural enrichments, pure cultures of (2) A. ferrooxidans and (3) A. thiooxidans were similar, but collectively differed from those of the natural heterotrophic enrichment cultures. The PLFA profiles for the heterotrophic communities were made up of primarily (88-99%) C16:0 and two isomers of C18:1. In contrast, the autotrophic communities had high proportions of C16:1 (up to 18%) as well as cyclo C17 and cyclo C19 PLFA that combined comprised 18 to 58% of the observed PLFA. The δ13C signatures of the PLFA also differed strongly between the two trophic levels. The δ13C of the autotrophic PLFA, - 24 to

  10. Survival of indicator organisms during enrichment on tetrachloroethene.

    Science.gov (United States)

    Skramstad, J D; Hurst, C J; Novak, P J

    2003-01-01

    A laboratory study was performed as the basis for a full-scale bioaugmentation project at a site contaminated with chlorinated ethenes. The objectives of this study were to 1) develop a protocol to enrich for a tetrachloroethene (PCE)-dechlorinating culture from waste activated sludge and anaerobic digester biosolids and 2) monitor the survival of fecal coliform bacteria and bacteriophage, which model enteric viruses, during the enrichment process. A culture was enriched in 8 days with the ability to degrade 6-microM PCE to cis-dichloroethene. Using the enrichment protocol in two identical experiments, significant inactivation of fecal coliform bacteria (2 log) and somatic coliphage (0.33 log) was observed in one of the experiments; no inactivation occurred in the second experiment. The number of F-specific coliphage decreased in both experiments (0.87 and 1.26 log inactivation). Despite the decrease in some of the coliform and bacteriophage numbers, the quantity of organisms and phage particles present after enrichment was still high (approximately 7.5 x 10(5) most probable number/L, 6.9 x 10(6) plaque-forming units (PFU)/L, and 3.3 x 10(5) PFU/L, for fecal coliform bacteria, somatic coliphage, and F-specific coliphage, respectively). This may be cause for concern, depending on the current and future groundwater use at or near a site undergoing bioaugmentation with cultures derived from waste activated sludge and anaerobic digester biosolids.

  11. mRNA differential display in a microbial enrichment culture: simultaneous identification of three cyclohexanone monooxygenases from three species.

    Science.gov (United States)

    Brzostowicz, Patricia C; Walters, Dana M; Thomas, Stuart M; Nagarajan, Vasantha; Rouvière, Pierre E

    2003-01-01

    mRNA differential display has been used to identify cyclohexanone oxidation genes in a mixed microbial community derived from a wastewater bioreactor. Thirteen DNA fragments randomly amplified from the total RNA of an enrichment subculture exposed to cyclohexanone corresponded to genes predicted to be involved in the degradation of cyclohexanone. Nine of these DNA fragments are part of genes encoding three distinct Baeyer-Villiger cyclohexanone monooxygenases from three different bacterial species present in the enrichment culture. In Arthrobacter sp. strain BP2 and Rhodococcus sp. strain Phi2, the monooxygenase is part of a gene cluster that includes all the genes required for the degradation of cyclohexanone, while in Rhodococcus sp. strain Phi1 the genes surrounding the monooxygenase are not predicted to be involved in this degradation pathway but rather seem to belong to a biosynthetic pathway. Furthermore, in the case of Arthrobacter strain BP2, three other genes flanking the monooxygenase were identified by differential display, demonstrating that the repeated sampling of bacterial operons shown earlier for a pure culture (D. M. Walters, R. Russ, H. Knackmuss, and P. E. Rouvière, Gene 273:305-315, 2001) is also possible for microbial communities. The activity of the three cyclohexanone monooxygenases was confirmed and characterized following their expression in Escherichia coli.

  12. Spontaneous bacterial peritonitis due to Listeria monocytogenes: importance of enrichment culture.

    Science.gov (United States)

    Jayasinghe, Saroj; Connor, Martin; Donaldson, Shona; Austin, Hannah; Foster, Adele

    2010-09-01

    A case of Listeria monocytogenes induced spontaneous bacterial peritonitis (SBP) is reported in a patient with primary biliary cirrhosis. It is an indolent illness and may not show a neutrophil reaction in peritoneal fluid. Enrichment broth was required to isolate L monocytogenes in the patient. This is not routinely used in the UK and therefore isolates may be missed. L monocytogenes remains sensitive to ampicillin, penicillin and gentamicin, but is resistant to cephalosporin antibiotics. The rising incidence of listeriosis in the population suggests that the incidence of SBP from L monocytogenes is likely to increase.

  13. Developments in techniques for the isolation, enrichment, main culture conditions and identification of spermatogonial stem cells

    OpenAIRE

    He, Yanan; Chen, Xiaoli; Zhu, Huabin; Wang, Dong

    2015-01-01

    The in vitro culture system of spermatogonial stem cells (SSCs) provides a basis for studies on spermatogenesis, and also contributes to the development of new methods for the preservation of livestock and animal genetic modification. In vitro culture systems have mainly been established for mouse SSCs, but are lacking for farm animals. We reviewed and analyzed the current progress in SSC techniques such as isolation, purification, cultivation and identification. Based on the published studie...

  14. Culturing bias in marine heterotrophic flagellates analyzed through seawater enrichment incubations.

    Science.gov (United States)

    del Campo, Javier; Balagué, Vanessa; Forn, Irene; Lekunberri, Itziar; Massana, Ramon

    2013-10-01

    The diversity of heterotrophic flagellates is generally based on cultivated strains, on which ultrastructural, physiological, and molecular studies have been performed. However, the relevance of these cultured strains as models of the dominant heterotrophic flagellates in the marine planktonic environment is unclear. In fact, molecular surveys typically recover novel eukaryotic lineages that have refused cultivation so far. This study was designed to directly address the culturing bias in planktonic marine heterotrophic flagellates. Several microcosms were established adding increasing amounts and sources of organic matter to a confined natural microbial community pre-filtered by 3 μm. Growth dynamics were followed by epifluorescence microscopy and showed the expected higher yield of bacteria and heterotrophic flagellates at increased organic matter additions. Moreover, protist diversity analyzed by molecular tools showed a clear substitution in the community, which differed more and more from the initial sample as the organic matter increased. Within this gradient, there was also an increase of sequences related to cultured organisms as well as a decrease in diversity. Culturing bias is partly explained by the use of organic matter in the isolation process, which drives a shift in the community to conditions closer to laboratory cultures. An intensive culturing effort using alternative isolation methods is necessary to allow the access to the missing heterotrophic flagellates that constitute the abundant and active taxa in marine systems.

  15. Succession of lignocellulolytic bacterial consortia bred anaerobically from lake sediment

    NARCIS (Netherlands)

    Korenblum, Elisa; Jiménez Avella, Diego; van Elsas, Jan

    2016-01-01

    Anaerobic bacteria degrade lignocellulose in various anoxic and organically rich environments, often in a syntrophic process. Anaerobic enrichments of bacterial communities on a recalcitrant lignocellulose source were studied combining polymerase chain reaction–denaturing gradient gel electrophoresi

  16. Applicability of a Lactobacillus amylovorus strain as co-culture for natural folate bio-enrichment of fermented milk.

    Science.gov (United States)

    Laiño, Jonathan Emiliano; Juarez del Valle, Marianela; Savoy de Giori, Graciela; LeBlanc, Jean Guy Joseph

    2014-11-17

    The ability of 55 strains from different Lactobacillus species to produce folate was investigated. In order to evaluate folic acid productivity, lactobacilli were cultivated in the folate-free culture medium (FACM). Most of the tested strains needed folate for growth. The production and the extent of vitamin accumulation were distinctive features of individual strains. Lactobacillus amylovorus CRL887 was selected for further studies because of its ability to produce significantly higher concentrations of vitamin (81.2 ± 5.4 μg/L). The safety of this newly identified folate producing strain was evaluated through healthy experimental mice. No bacterial translocation was detected in liver and spleen after consumption of CRL887 during 7 days and no undesirable side effects were observed in the animals that received this strain. This strain in co-culture with previously selected folate producing starter cultures (Lactobacillus bulgaricus CRL871, and Streptococcus thermophilus CRL803 and CRL415) yielded a yogurt containing high folate concentrations (263.1 ± 2.4 μg/L); a single portion of which would provide 15% of the recommended dietary allowance. This is the first report where a Lactobacillus amylovorus strain was successfully used as co-culture for natural folate bio-enrichment of fermented milk.

  17. Reduction of Salmonella Typhimurium by Fermentation Metabolites of Diamond V Original XPC in an In Vitro Anaerobic Mixed Chicken Cecal Culture

    Science.gov (United States)

    Rubinelli, Peter; Roto, Stephanie; Kim, Sun Ae; Park, Si Hong; Pavlidis, Hilary O.; McIntyre, Don; Ricke, Steven C.

    2016-01-01

    Fermentation metabolites of Diamond V Original XPC™ (XPC), a biological product derived from yeast fermentation, were evaluated for their ability to reduce the Salmonella Typhimurium population using an in vitro mixed anaerobic culture system containing cecal microbiota to simulate chicken hindgut conditions. Four different samples were prepared: anaerobic mixed culture containing (1) feed only, (2) cecal only (ceca were harvested from 42 days old broiler chickens), (3) feed and cecal contents, and (4) feed, cecal contents, and 1% XPC. Two experimental conditions were investigated: Group 1, in which the cecal content was added at the same time as a S. Typhimurium marker strain and Group 2, in which the cecal content was preincubated for 24 h prior to the inoculation with the S. Typhimurium marker strain. The mixed cultures were incubated anaerobically at 37°C, and the S. Typhimurium marker strain was enumerated at 0, 24, and 48 h. Analysis of short chain fatty acids was also conducted for 24 h. In the Group 1 experiment, adding XPC did not exhibit significant reduction of S. Typhimurium. However, the presence of XPC resulted in rapid reduction of S. Typhimurium in Group 2. S. Typhimurium was reduced from 6.81 log10 CFU/ml (0 h) to 3.73 log10 CFU/ml and 1.19 log10 CFU/ml after 24 and 48 h, respectively. These levels were also 2.47 log10 and 2.72 log10 lower than the S. Typhimurium level recovered from the control culture with feed and cecal contents, but without XPC. Based on these results, it appears that the ability of XPC to reduce S. Typhimurium requires the presence of the cecal microbiota. Short chain fatty acid analysis indicated that acetate and butyrate concentrations of cultures containing XPC were twofold greater than the control cultures by 24 h of anaerobic growth. Results from the present study suggest that dietary inclusion of XPC may influence cecal microbiota fermentation and has the potential to reduce Salmonella in the cecum

  18. Atrazine biodegradation efficiency, metabolite detection, and trzD gene expression by enrichment bacterial cultures from agricultural soil.

    Science.gov (United States)

    Solomon, Robinson David Jebakumar; Kumar, Amit; Satheeja Santhi, Velayudhan

    2013-12-01

    Atrazine is a selective herbicide used in agricultural fields to control the emergence of broadleaf and grassy weeds. The persistence of this herbicide is influenced by the metabolic action of habituated native microorganisms. This study provides information on the occurrence of atrazine mineralizing bacterial strains with faster metabolizing ability. The enrichment cultures were tested for the biodegradation of atrazine by high-performance liquid chromatography (HPLC) and mass spectrometry. Nine cultures JS01.Deg01 to JS09.Deg01 were identified as the degrader of atrazine in the enrichment culture. The three isolates JS04.Deg01, JS07.Deg01, and JS08.Deg01 were identified as efficient atrazine metabolizers. Isolates JS04.Deg01 and JS07.Deg01 produced hydroxyatrazine (HA) N-isopropylammelide and cyanuric acid by dealkylation reaction. The isolate JS08.Deg01 generated deethylatrazine (DEA), deisopropylatrazine (DIA), and cyanuric acid by N-dealkylation in the upper degradation pathway and later it incorporated cyanuric acid in their biomass by the lower degradation pathway. The optimum pH for degrading atrazine by JS08.Deg01 was 7.0 and 16S rDNA phylogenetic typing identified it as Enterobacter cloacae strain JS08.Deg01. The highest atrazine mineralization was observed in case of isolate JS08.Deg01, where an ample amount of trzD mRNA was quantified at 72 h of incubation with atrazine. Atrazine bioremediating isolate E. cloacae strain JS08.Deg01 could be the better environmental remediator of agricultural soils and the crop fields contaminated with atrazine could be the source of the efficient biodegrading microbial strains for the environmental cleanup process.

  19. Atrazine biodegradation efficiency, metabolite detection, and trzD gene expression by enrichment bacterial cultures from agricultural soil

    Institute of Scientific and Technical Information of China (English)

    Robinson David Jebakumar SOLOMON; Amit KUMAR; Velayudhan SATHEEJA SANTHI

    2013-01-01

    Atrazine is a selective herbicide used in agricultural fields to control the emergence of broadleaf and grassy weeds. The persistence of this herbicide is influenced by the metabolic action of habituated native microor-ganisms. This study provides information on the occurrence of atrazine mineralizing bacterial strains with faster me-tabolizing ability. The enrichment cultures were tested for the biodegradation of atrazine by high-performance liquid chromatography (HPLC) and mass spectrometry. Nine cultures JS01.Deg01 to JS09.Deg01 were identified as the degrader of atrazine in the enrichment culture. The three isolates JS04.Deg01, JS07.Deg01, and JS08.Deg01 were identified as efficient atrazine metabolizers. Isolates JS04.Deg01 and JS07.Deg01 produced hydroxyatrazine (HA) N-isopropylammelide and cyanuric acid by dealkylation reaction. The isolate JS08.Deg01 generated deethylatrazine (DEA), deisopropylatrazine (DIA), and cyanuric acid by N-dealkylation in the upper degradation pathway and later it incorporated cyanuric acid in their biomass by the lower degradation pathway. The optimum pH for degrading atrazine by JS08.Deg01 was 7.0 and 16S rDNA phylogenetic typing identified it as Enterobacter cloacae strain JS08.Deg01. The highest atrazine mineralization was observed in case of isolate JS08.Deg01, where an ample amount of trzD mRNA was quantified at 72 h of incubation with atrazine. Atrazine bioremediating isolate E. cloacae strain JS08.Deg01 could be the better environmental remediator of agricultural soils and the crop fields contaminated with atrazine could be the source of the efficient biodegrading microbial strains for the environmental cleanup process.

  20. Lactate induces tumour necrosis factor-alpha, interleukin-6 and interleukin-1beta release in microglial- and astroglial-enriched primary cultures.

    Science.gov (United States)

    Andersson, Anna K; Rönnbäck, Lars; Hansson, Elisabeth

    2005-06-01

    Hyperammonaemia has deleterious effects on the CNS in patients with liver dysfunction. Cellular mechanisms underlying the effects of hyperammonaemia are largely unknown, although astrocytes have been the main target of interest. This study investigated how treatment with NH4Cl and lactate, which increase in the brain as a consequence of hyperammonaemia, affects cells in primary rat cultures enriched in either astrocytes or microglia. Morphological changes were studied over time using light microscopy. Release of the proinflammatory cytokines tumour necrosis factor-alpha (TNF-alpha), interleukin (IL)-6 and IL-1beta was measured using ELISA. NH4Cl was found to induce vacuole formation in both culture systems. Lactate treatment altered astrocytic appearance, resulting in increased space between individual cells. Microglia adopted a round morphology with either NH4Cl or lactate treatment. Lactate, but not NH4Cl, induced release of TNF-alpha and IL-6 in both astroglial- and microglial-enriched cultures, while IL-1beta was released only in microglial cultures. Cytokine release was higher in the microglial- than in the astroglial-enriched cultures. Additionally, the astroglial-enriched cultures containing approximately 10% microglial cells released more cytokines than cultures containing about 5% microglial cells. Taken together, our data suggest that most TNF-alpha, IL-6 and IL-1beta release comes from microglia. Thus, microglia could play an important role in the pathological process of hyperammonaemia.

  1. Anaerobic Biodegradation of Tetrachloroethylene with Acetic Acid as Cometabolism Substrate under Anaerobic Condition

    Institute of Scientific and Technical Information of China (English)

    LI Ye; LIU Fei; CHEN Honghan; SHI Jinhua; WANG Yufan

    2008-01-01

    A series of batch-type experiments with acetate acid as the primary substrate wereperformed using enrichment cultures developed from the anaerobic sludge to investigate the effect ofacetate acid on tetrachloroethylene (PCE) biodegradation. Experimental results indicated that acetateacid was an efficient electron donor in affecting the biotransformability of PCE. Trichloroethylene(TCE) was the primary dehalogenation product, and small amounts of dichloroethylenes (DCEs) werealso detected. No significant further DCEs degradation was detected. PCE degradation rate in theexperiment was 36.6 times faster than background rate in natural groundwater.

  2. One health from a social-ecological systems perspective: enriching social and cultural dimensions.

    Science.gov (United States)

    Ross, Helen

    2013-01-01

    This chapter offers insights from the environmental management paradigm of 'social-ecological systems' and related bodies of theory on people-environment relationships to assist the evolution of the One Health interdisciplinary endeavour of health promotion across human-animal ecosystem relationships. It also seeks to expand thinking about the social and cultural dimensions that are likely to prove important in the development of thinking and practice in the One Health field. It advocates consideration of cultural and economic relationships affecting people's interactions with domesticated and wild animal species and ecosystems, and exploration of the cognitive and behavioural aspects of these interactions.

  3. Biodegradation of Chlorpyrifos by Pseudomonas Resinovarans Strain AST2.2 Isolated from Enriched Cultures.

    Directory of Open Access Journals (Sweden)

    Anish Sharma*,

    2016-04-01

    Full Text Available A bacterial strain AST2.2 with chlorpyrifos degrading ability was isolated by enrichment technique from apple orchard soil with previous history of chlorpyrifos use. Based on the morphological, biochemical tests and 16S rRNA sequence analysis, AST2.2 strain was identified as Pseudomonas resinovarans. The strain AST2.2 utilized chlorpyrifos as the sole source of carbon and energy. This strain exhibited growth upto 400mg/l concentration of chlorpyrifos and exhibited high extracellular organophosphorus hydrolase (OPH activity. Gas chromatography-flame ionization detector (GC-FID studies revealed that Pseudomonas resinovarans AST2.2 degraded 43.90 % of chlorpyrifos (400 mg/l within 96 hrs. Intermediates of chlorpyrifos degradation were identified using GC-MS. This strain have potential to degrade chlorpyrifos and thus can be used for bioremediation and ecological restoration of sites contaminated with chlorpyrifos.

  4. Real-time PCR detection of Campylobacter spp.: A comparison toclassic culturing and enrichment

    NARCIS (Netherlands)

    Boer, P. de; Rahaoui, H.; Leer, R.J.; Montijn, R.C.; Vossen, J.M.B.M. van der

    2015-01-01

    The major disadvantage of the current gold standard for detection of the food pathogen Campylobacter, i.e. culturing, is the lengthy procedure. In this study we assessed the use of real-time PCR for detection of Campylobacter. To this end, 926 poultry samples, taken from transport containers and bro

  5. Culturally Diverse Literature: Enriching Variety in an Era of Common Core State Standards

    Science.gov (United States)

    Boyd, Fenice B.; Causey, Lauren L.; Galda, Lee

    2015-01-01

    The authors argue for the overwhelming importance of finding and including culturally diverse literature into the curricula teachers are authorized to teach. They discuss the implications of use and offer ideas on how to identify quality literature to include in classroom and school libraries.

  6. Effects of Culture Media and Light Intensity on in vitro Growth of Oncidium under CO2 Enrichment Condition

    Institute of Scientific and Technical Information of China (English)

    He Songlin; Pan Huitang; Yang Qiusheng; Kong Dezheng; Zhang Qixiang; Michio Tanaka

    2003-01-01

    The effects of culture media and light intensity on in vitro growth of Oncidium 'Aloha Iwanga' were investigated under CO2 enrichment condition. Height, fresh and dry weight of the Oncidium seedlings were measured, and the leaf number per plant, shoot number per plant, leaf width and leaf chlorophyll content were also investigated. The results were as follows: 1) The seedling height, fresh and dry weight, leaf number per plant, leaf width and leaf chlorophyll content of the shoots growing on MS complete culture medium were higher than those on 1/2MS, VW and 1/2VW media. The root number per plant and ratio of dry matter of the seedlings cultured on 1/2MS and 1/2VW media were higher than those on MS and VW; 2) The seedling height, fresh weight, dry weight, dry matter ratio and leaf chlorophyll content, leaf length, leaf width, root length, leaf number per plant, root number per plant of seedlings of Oncidium growing under 4 500 lx and 1 700 lx were higher than those under 750 lx. However, there was no significant difference in those growth parameters mentioned above while dealing with 4 500 lx and 1 700 lx except for the seedling height. Nevertheless, the leaf color of plants under 4 500 lx was lighter and the leaves of the lower parts became yellowish in comparison with those growing under 1 700 lx.

  7. Stable acetate production in extreme-thermophilic (70°C) mixed culture fermentation by selective enrichment of hydrogenotrophic methanogens

    Science.gov (United States)

    Zhang, Fang; Zhang, Yan; Ding, Jing; Dai, Kun; van Loosdrecht, Mark C. M.; Zeng, Raymond J.

    2014-06-01

    The control of metabolite production is difficult in mixed culture fermentation. This is particularly related to hydrogen inhibition. In this work, hydrogenotrophic methanogens were selectively enriched to reduce the hydrogen partial pressure and to realize efficient acetate production in extreme-thermophilic (70°C) mixed culture fermentation. The continuous stirred tank reactor (CSTR) was stable operated during 100 days, in which acetate accounted for more than 90% of metabolites in liquid solutions. The yields of acetate, methane and biomass in CSTR were 1.5 +/- 0.06, 1.0 +/- 0.13 and 0.4 +/- 0.05 mol/mol glucose, respectively, close to the theoretical expected values. The CSTR effluent was stable and no further conversion occurred when incubated for 14 days in a batch reactor. In fed-batch experiments, acetate could be produced up to 34.4 g/L, significantly higher than observed in common hydrogen producing fermentations. Acetate also accounted for more than 90% of soluble products formed in these fed-batch fermentations. The microbial community analysis revealed hydrogenotrophic methanogens (mainly Methanothermobacter thermautotrophicus and Methanobacterium thermoaggregans) as 98% of Archaea, confirming that high temperature will select hydrogenotrophic methanogens over aceticlastic methanogens effectively. This work demonstrated a potential application to effectively produce acetate as a value chemical and methane as an energy gas together via mixed culture fermentation.

  8. Enhanced biogas yield from energy crops with rumen anaerobic fungi

    Energy Technology Data Exchange (ETDEWEB)

    Prochazka, Jindrich; Zabranska, Jana; Dohanyos, Michal [Department of Water Technology and Environmental Engineering, Faculty of Environmental Technology, Institute of Chemical Technology in Prague, Prague (Czech Republic); Mrazek, Jakub; Strosova, Lenka; Fliegerova, Katerina [Laboratory of Anaerobic Microbiology, Institute of Animal Physiology and Genetics, CAS, v.v.i., Prague (Czech Republic)

    2012-06-15

    Anaerobic fungi (AF) are able to degrade crop substrates with higher efficiency than commonly used anaerobic bacteria. The aim of this study was to investigate ways of use of rumen AF to improve biogas production from energy crops under laboratory conditions. In this study, strains of AF isolated from feces or rumen fluid of cows and deer were tested for their ability to integrate into the anaerobic bacterial ecosystem used for biogas production, in order to improve degradation of substrate polysaccharides and consequently the biogas yield. Batch culture, fed batch culture, and semicontinuous experiments have been performed using anaerobic sludge from pig slurry fermentation and different kinds of substrates (celluloses, maize, and grass silage) inoculated by different genera of AF. All experiments showed a positive effect of AF on the biogas yield and quality. AF improved the biogas production by 4-22%, depending on the substrate and AF species used. However, all the cultivation experiments indicated that rumen fungi do not show long-term survival in fermenters with digestate from pig slurry. The best results were achieved during fed batch experiment with fungal culture Anaeromyces (KF8), in which biogas production was enhanced during the whole experimental period of 140 days. This result has not been achieved in semicontinuous experiment, where increment in biogas production in fungal enriched reactor was only 4% after 42 days. (copyright 2012 Wiley-VCH Verlag GmbH and Co. KGaA, Weinheim)

  9. Nicotine suppresses lipopolysaccharide-induced release of interleukin-6 in mixed glia and microglia-enriched cultures

    Institute of Scientific and Technical Information of China (English)

    Zhihua Li; Qingzan Zhao; Hua Zhang; Xiuhua Ren; Mingfu Zhou; Weidong Zang

    2011-01-01

    Inflammation plays an important role in the pathogenesis of Parkinson's disease (PD) through the over-activation of microglia.Epidemiological studies show that smoking is associated with a lower incidence of PD.This study hypothesized that the neuroprotective effect of nicotine is mediated by modulating the activation of microglia via cytokine release.This study found that nicotine pretreatment suppressed the lipopolysaccharide-induced inflammatory reaction in the nervous system, especially microglia activation and interleukin-6 production.The inhibitory effects of 100 pmol/L nicotine were stronger compared with 1 and 10 pmol/L nicotine.These findings indicate that nicotine significantly decreases the production of proinflammatory interleukin-6 in mixed glia or microglia-enriched cultures, and plays an inhibitory effect on the lipopolysaccharide-induced inflammatory reaction.

  10. Modeling microbial ethanol production by E. coli under aerobic/anaerobic conditions: applicability to real postmortem cases and to postmortem blood derived microbial cultures.

    Science.gov (United States)

    Boumba, Vassiliki A; Kourkoumelis, Nikolaos; Gousia, Panagiota; Economou, Vangelis; Papadopoulou, Chrissanthy; Vougiouklakis, Theodore

    2013-10-10

    The mathematical modeling of the microbial ethanol production under strict anaerobic experimental conditions for some bacterial species has been proposed by our research group as the first approximation to the quantification of the microbial ethanol production in cases where other alcohols were produced simultaneously with ethanol. The present study aims to: (i) study the microbial ethanol production by Escherichia coli under controlled aerobic/anaerobic conditions; (ii) model the correlation between the microbial produced ethanol and the other higher alcohols; and (iii) test their applicability in: (a) real postmortem cases that had positive BACs (>0.10 g/L) and co-detection of higher alcohols and 1-butanol during the original ethanol analysis and (b) postmortem blood derived microbial cultures under aerobic/anaerobic controlled experimental conditions. The statistical evaluation of the results revealed that the formulated models were presumably correlated to 1-propanol and 1-butanol which were recognized as the most significant descriptors of the modeling process. The significance of 1-propanol and 1-butanol as descriptors was so powerful that they could be used as the only independent variables to create a simple and satisfactory model. The current models showed a potential for application to estimate microbial ethanol - within an acceptable standard error - in various tested cases where ethanol and other alcohols have been produced from different microbes.

  11. Anaerobic Biotransformation and Mobility of Pu and of Pu-EDTA

    Energy Technology Data Exchange (ETDEWEB)

    Xun, Luying

    2009-11-20

    The enhanced mobility of radionuclides by co-disposed chelating agent, ethylenediaminetetraacetate (EDTA), is likely to occur only under anaerobic conditions. Our extensive effort to enrich and isolate anaerobic EDTA-degrading bacteria has failed. Others has tried and also failed. To explain the lack of anaerobic biodegradation of EDTA, we proposed that EDTA has to be transported into the cells for metabolism. A failure of uptake may contribute to the lack of EDTA degradation under anaerobic conditions. We demonstrated that an aerobic EDTA-degrading bacterium strain BNC1 uses an ABC-type transporter system to uptake EDTA. The system has a periplasmic binding protein that bind EDTA and then interacts with membrane proteins to transport EDTA into the cell at the expense of ATP. The bind protein EppA binds only free EDTA with a Kd of 25 nM. The low Kd value indicates high affinity. However, the Kd value of Ni-EDTA is 2.4 x 10^(-10) nM, indicating much stronger stability. Since Ni and other trace metals are essential for anaerobic respiration, we conclude that the added EDTA sequestrates all trace metals and making anaerobic respiration impossible. Thus, the data explain the lack of anaerobic enrichment cultures for EDTA degradation. Although we did not obtain an EDTA degrading culture under anaerobic conditions, our finding may promote the use of certain metals that forms more stable metal-EDTA complexes than Pu(III)-EDTA to prevent the enhanced mobility. Further, our data explain why EDTA is the most dominant organic pollutant in surface waters, due to the lack of degradation of certain metal-EDTA complexes.

  12. Real-time PCR detection of Campylobacter spp.: A comparison to classic culturing and enrichment.

    Science.gov (United States)

    de Boer, P; Rahaoui, H; Leer, R J; Montijn, R C; van der Vossen, J M B M

    2015-10-01

    The major disadvantage of the current gold standard for detection of the food pathogen Campylobacter, i.e. culturing, is the lengthy procedure. In this study we assessed the use of real-time PCR for detection of Campylobacter. To this end, 926 poultry samples, taken from transport containers and broiler caeca in The Netherlands in 2007, were subjected to three different real-time PCR detection methods: one targeting the Campylobacter jejuni hipO gene, one targeting the Campylobacter coli glyA gene, and one generically targeting Campylobacter spp. 16S rDNA sequence. The PCR results from the three different PCR protocols were compared to the work of Nauta et al. (2009) who analyzed the same set of samples collected from 62 broiler flocks by means of enrichment culturing. The results indicate that the generic 16S campylobacter PCR detection is equally reliable but much faster (4 h instead of ≥2 days) than detection by means of culturing. Moreover, PCR detection targeting the hipO and the glyA gene provide the possibility of C. jejuni and C. coli species discrimination. The generic Campylobacter spp. PCR analysis also confirmed the high incidence of Campylobacter spp. in poultry samples (∼90%) and the species specific PCR showed the simultaneous presence of C. jejuni and C. coli in ∼24% of the samples. Furthermore, the results from the three PCR analyses suggested the occurrence of alternative Campylobacter species in almost 10% of the samples. The campylobacter PCR detection methods reported here can replace traditional culturing because of being quicker and more reliable.

  13. Predominance of Viable Spore-Forming Piezophilic Bacteria in High-Pressure Enrichment Cultures from ~1.5 to 2.4 km-Deep Coal-Bearing Sediments below the Ocean Floor

    Science.gov (United States)

    Fang, Jiasong; Kato, Chiaki; Runko, Gabriella M.; Nogi, Yuichi; Hori, Tomoyuki; Li, Jiangtao; Morono, Yuki; Inagaki, Fumio

    2017-01-01

    Phylogenetically diverse microorganisms have been observed in marine subsurface sediments down to ~2.5 km below the seafloor (kmbsf). However, very little is known about the pressure-adapted and/or pressure-loving microorganisms, the so called piezophiles, in the deep subseafloor biosphere, despite that pressure directly affects microbial physiology, metabolism, and biogeochemical processes of carbon and other elements in situ. In this study, we studied taxonomic compositions of microbial communities in high-pressure incubated sediment, obtained during the Integrated Ocean Drilling Program (IODP) Expedition 337 off the Shimokita Peninsula, Japan. Analysis of 16S rRNA gene-tagged sequences showed that members of spore-forming bacteria within Firmicutes and Actinobacteria were predominantly detected in all enrichment cultures from ~1.5 to 2.4 km-deep sediment samples, followed by members of Proteobacteria, Acidobacteria, and Bacteroidetes according to the sequence frequency. To further study the physiology of the deep subseafloor sedimentary piezophilic bacteria, we isolated and characterized two bacterial strains, 19R1-5 and 29R7-12, from 1.9 and 2.4 km-deep sediment samples, respectively. The isolates were both low G+C content, gram-positive, endospore-forming and facultative anaerobic piezophilic bacteria, closely related to Virgibacillus pantothenticus and Bacillus subtilis within the phylum Firmicutes, respectively. The optimal pressure and temperature conditions for growth were 20 MPa and 42°C for strain 19R1-5, and 10 MPa and 43°C for strain 29R7-12. Bacterial (endo)spores were observed in both the enrichment and pure cultures examined, suggesting that these piezophilic members were derived from microbial communities buried in the ~20 million-year-old coal-bearing sediments after the long-term survival as spores and that the deep biosphere may host more abundant gram-positive spore-forming bacteria and their spores than hitherto recognized. PMID:28220112

  14. The effects of theaflavin-enriched black tea extract on muscle soreness, oxidative stress, inflammation, and endocrine responses to acute anaerobic interval training: a randomized, double-blind, crossover study

    Directory of Open Access Journals (Sweden)

    Golem Devon L

    2010-02-01

    Full Text Available Abstract Background Muscle soreness and decreased performance often follow a bout of high-intensity exercise. By reducing these effects, an athlete can train more frequently and increase long-term performance. The purpose of this study is to examine whether a high-potency, black tea extract (BTE alters the delayed onset muscle soreness (DOMS, oxidative stress, inflammation, and cortisol (CORT responses to high-intensity anaerobic exercise. Methods College-age males (N = 18 with 1+ yrs of weight training experience completed a double-blind, placebo-controlled, crossover study. Subjects consumed the BTE (1,760 mg BTE·d-1 or placebo (PLA for 9 days. Each subject completed two testing sessions (T1 & T2, which occurred on day 7 of the intervention. T1 & T2 consisted of a 30 s Wingate Test plus eight 10 s intervals. Blood samples were obtained before, 0, 30 & 60 min following the interval sessions and were used to analyze the total to oxidized glutathione ratio (GSH:GSSG, 8-isoprostane (8-iso, CORT, and interleukin 6 (IL-6 secretion. DOMS was recorded at 24 & 48 h post-test using a visual analog scale while BTE or PLA continued to be administered. Significance was set at P . Results Compared to PLA, BTE produced significantly higher average peak power (P = 0.013 and higher average mean power (P = 0.067 across nine WAnT intervals. BTE produced significantly lower DOMS compared to PLA at 24 h post test (P and 48 h post test (P . Compared to PLA, BTE had a slightly higher GSH:GSSG ratio at baseline which became significantly higher at 30 and 60 min post test (P . AUC analysis revealed BTE to elicit significantly lower GSSG secretion (P = 0.009, significantly higher GSH:GSSG ratio (P = 0.001, and lower CORT secretion (P = 0.078 than PLA. AUC analysis did not reveal a significant difference in total IL-6 response (P = 0.145 between conditions. Conclusions Consumption of theaflavin-enriched black tea extract led to improved recovery and a reduction in

  15. The effects of theaflavin-enriched black tea extract on muscle soreness, oxidative stress, inflammation, and endocrine responses to acute anaerobic interval training: a randomized, double-blind, crossover study

    Science.gov (United States)

    2010-01-01

    Background Muscle soreness and decreased performance often follow a bout of high-intensity exercise. By reducing these effects, an athlete can train more frequently and increase long-term performance. The purpose of this study is to examine whether a high-potency, black tea extract (BTE) alters the delayed onset muscle soreness (DOMS), oxidative stress, inflammation, and cortisol (CORT) responses to high-intensity anaerobic exercise. Methods College-age males (N = 18) with 1+ yrs of weight training experience completed a double-blind, placebo-controlled, crossover study. Subjects consumed the BTE (1,760 mg BTE·d-1) or placebo (PLA) for 9 days. Each subject completed two testing sessions (T1 & T2), which occurred on day 7 of the intervention. T1 & T2 consisted of a 30 s Wingate Test plus eight 10 s intervals. Blood samples were obtained before, 0, 30 & 60 min following the interval sessions and were used to analyze the total to oxidized glutathione ratio (GSH:GSSG), 8-isoprostane (8-iso), CORT, and interleukin 6 (IL-6) secretion. DOMS was recorded at 24 & 48 h post-test using a visual analog scale while BTE or PLA continued to be administered. Significance was set at P < 0.05. Results Compared to PLA, BTE produced significantly higher average peak power (P = 0.013) and higher average mean power (P = 0.067) across nine WAnT intervals. BTE produced significantly lower DOMS compared to PLA at 24 h post test (P < 0.001) and 48 h post test (P < 0.001). Compared to PLA, BTE had a slightly higher GSH:GSSG ratio at baseline which became significantly higher at 30 and 60 min post test (P < 0.002). AUC analysis revealed BTE to elicit significantly lower GSSG secretion (P = 0.009), significantly higher GSH:GSSG ratio (P = 0.001), and lower CORT secretion (P = 0.078) than PLA. AUC analysis did not reveal a significant difference in total IL-6 response (P = 0.145) between conditions. Conclusions Consumption of theaflavin-enriched black tea extract led to improved recovery and a

  16. Cellular responses of Saccharomyces cerevisiae at near-zero growth rates: transcriptome analysis of anaerobic retentostat cultures

    NARCIS (Netherlands)

    Boender, L.G.M.; Van Maris, A.J.A.; De Hulster, E.A.F.; Almering, M.J.H.; Van der Klei, I.J.; Veenhuis, M.; De Winde, J.H.; Pronk, J.T.; Daran-Lapujade, P.A.S.

    2011-01-01

    Extremely low specific growth rates (below 0.01 h−1) represent a largely unexplored area of microbial physiology. In this study, anaerobic, glucose-limited retentostats were used to analyse physiological and genome-wide transcriptional responses of Saccharomyces cerevisiae to cultivation at near-zer

  17. Cellular responses of Saccharomyces cerevisiae at near-zero growth rates : transcriptome analysis of anaerobic retentostat cultures

    NARCIS (Netherlands)

    Boender, Leonie G. M.; van Maris, Antonius J. A.; de Hulster, Erik A. F.; Almering, Marinka J. H.; van der Klei, Ida J.; Veenhuis, Marten; de Winde, Johannes H.; Pronk, Jack T.; Daran-Lapujade, Pascale

    2011-01-01

    Extremely low specific growth rates (below 0.01 h(-1)) represent a largely unexplored area of microbial physiology. In this study, anaerobic, glucose-limited retentostats were used to analyse physiological and genome-wide transcriptional responses of Saccharomyces cerevisiae to cultivation at near-z

  18. A discourse on creating and rendering educational and cultural enrichment services to rural migrant workers by public libraries

    Institute of Scientific and Technical Information of China (English)

    WANG; Zizhou; Charles; C.Yan

    2009-01-01

    As far as China’s enormous success in her economic reforms and development is concerned,Chinese rural migrant workers indeed have had their biggest share of contributions.Yet ironically in return,they have only received a disproportionate meager share of benefit rewards.These people represent a huge economically deprived group at the bottom of the social totem pole in China’s metropolises.On the whole as a social group,their educational attainment is relatively low as compared to their average urban coworkers.As such being the case,their rights to access some of the cost-free cultural and educational enrichment programs in the society are limited and not always assured.Nevertheless and in general speaking,they manifest a strong and consistent desire to acquire all sorts of new and practical knowledge by means of accessing the resources and facilities of their local public libraries.It is suggested in this paper that public libraries are in a good position to give strong support to the central government’s strategic planning for the development of a public culture service systema).In implementing such a government initiative with an unswerving purpose of advancing social justice and equality,public libraries should strife to provide as many as possible their library services at no cost to the public,especially to those socially deprived rural migrant workers.

  19. Implants composed of carbon fiber mesh and bone-marrow-derived, chondrocyte-enriched cultures for joint surface reconstruction.

    Science.gov (United States)

    Robinson, D; Efrat, M; Mendes, D G; Halperin, N; Nevo, Z

    1993-01-01

    The current study integrates two distinct approaches in joint resurfacing into a combined type of implant, composed of carbon fiber mesh impregnated and coated with a hyaluronic-acid-based delivery substance containing cultured cells. Rabbit autogeneic chondrocyte-enriched cultures obtained from mesenchymal stem cells (chondroprogenitor cells) derived from adult rabbit bone marrow were grown in vitro under conditions favoring chondrogenesis. The improvement in quality of repair when a combined implant containing both cells and a carbon scaffold was used, in comparison to the utilization of carbon fiber mesh alone, was clearly demonstrated using clinical, histological, biochemical, and biomechanical examinations. Evaluations of the joints were performed at 6 weeks and 6 months after implantation. The repair tissue in the cell-implanted joints consisted of a typical hyaline cartilage, which was more cellular and thicker than the repair tissue in the hyaluronic-acid-impregnated carbon-fiber-implanted control joints. The hyaline cartilage in the experimental group formed a superficial layer above the carbon fibers, flush with the joint surface. In the controls, in which carbon fiber and the delivery substance alone were implanted, a histologically and biochemically fibrous tissue that was inferior biomechanically to the new cartilage was formed by the cells containing implants.

  20. Enhancement of Phenolics, Resveratrol and Antioxidant Activity by Nitrogen Enrichment in Cell Suspension Culture of Vitis vinifera

    Directory of Open Access Journals (Sweden)

    Napaporn Sae-Lee

    2014-06-01

    Full Text Available Ammonium nitrate (NH4NO3, an important nitrogen source (34% N, has been used as an elicitor to stimulate plant growth and development as well as induce secondary metabolites under controlled conditions. In the present paper, we investigated the enhancement of cell biomass, total phenolics, resveratrol levels, and antioxidant activity of Vitis vinifera cv. Pok Dum by nitrogen enrichment (MS medium supplemented with NH4NO3 at 0, 500, 1,000, 5,000 and 10,000 mg/L. The highest accumulations of biomass, phenolics and resveratrol contents were observed at 8.8-fold (86.6 g DW/L, 15.9-fold (71.91 mg GAE/g DW and 5.6-fold (277.89 µg/g DW by the 14th day, in the medium supplemented with 500 mg/L NH4NO3. Moreover, the antioxidant activities of cultured grape cells estimated by the DPPH· and ABTS·+ assay were positively correlated with phenolics and resveratrol, and the maximum activity was also observed in cultured cells with 500 mg/L NH4NO3 at 176.11 and 267.79 mmol TE/100 g DW, respectively.

  1. Characterization of Fe (III)-reducing enrichment culture and isolation of Fe (III)-reducing bacterium Enterobacter sp. L6 from marine sediment.

    Science.gov (United States)

    Liu, Hongyan; Wang, Hongyu

    2016-07-01

    To enrich the Fe (III)-reducing bacteria, sludge from marine sediment was inoculated into the medium using Fe (OH)3 as the sole electron acceptor. Efficiency of Fe (III) reduction and composition of Fe (III)-reducing enrichment culture were analyzed. The results indicated that the Fe (III)-reducing enrichment culture with the dominant bacteria relating to Clostridium and Enterobacter sp. had high Fe (III) reduction of (2.73 ± 0.13) mmol/L-Fe (II). A new Fe (III)-reducing bacterium was isolated from the Fe (III)-reducing enrichment culture and identified as Enterobacter sp. L6 by 16S rRNA gene sequence analysis. The Fe (III)-reducing ability of strain L6 under different culture conditions was investigated. The results indicated that strain L6 had high Fe (III)-reducing activity using glucose and pyruvate as carbon sources. Strain L6 could reduce Fe (III) at the range of NaCl concentrations tested and had the highest Fe (III) reduction of (4.63 ± 0.27) mmol/L Fe (II) at the NaCl concentration of 4 g/L. This strain L6 could reduce Fe (III) with unique properties in adaptability to salt variation, which indicated that it can be used as a model organism to study Fe (III)-reducing activity isolated from marine environment.

  2. Characterization of bacterial diversity in an atrazine degrading enrichment culture and degradation of atrazine, cyanuric acid and biuret in industrial wastewater.

    Science.gov (United States)

    Dutta, Anirban; Vasudevan, Venugopal; Nain, Lata; Singh, Neera

    2016-01-01

    An enrichment culture was used to study atrazine degradation in mineral salt medium (MSM) (T1), MSM+soil extract (1:1, v/v) (T2) and soil extract (T3). Results suggested that enrichment culture required soil extract to degrade atrazine, as after second sequential transfer only partial atrazine degradation was observed in T1 treatment while atrazine was completely degraded in T2 and T3 treatments even after fourth transfer. Culture independent polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE) technique confirmed selective enrichment of genus Bacillus along with Pseudomonas and Burkholderia. Degradation of atrazine/metabolites in the industrial wastewater was studied at different initial concentrations of the contaminants [wastewater-water (v/v) ratio: T1, 1:9; T2, 2:8; T3, 3:7; T4, 5:5 and T5, undiluted effluent]. The initial concentrations of atrazine, cyanuric acid and biuret ranged between 5.32 and 53.92 µg mL(-1), 265.6 and 1805.2 µg mL(-1) and 1.85 and 16.12 µg mL(-1), respectively. The enrichment culture was able to completely degrade atrazine, cyanuric acid and biuret up to T4 treatment, while no appreciable degradation of contaminants was observed in the undiluted effluent (T5). Inability of enrichment culture to degrade atrazine/metabolites might be due to high concentrations of cyanuric acid. Therefore, a separate study on cyanuric acid degradation suggested: (i) no appreciable cyanuric acid degradation with accumulation of an unidentified metabolite in the medium where cyanuric acid was supplemented as the sole source of carbon and nitrogen; (ii) partial cyanuric acid degradation with accumulation of unidentified metabolite in the medium containing additional nitrogen source; and (iii) complete cyanuric acid degradation in the medium supplemented with an additional carbon source. This unidentified metabolite observed during cyanuric acid degradation and also detected in the enrichment culture inoculated wastewater samples

  3. Establishment of microbial eukaryotic enrichment cultures from a chemically stratified antarctic lake and assessment of carbon fixation potential.

    Science.gov (United States)

    Dolhi, Jenna M; Ketchum, Nicholas; Morgan-Kiss, Rachael M

    2012-04-20

    Lake Bonney is one of numerous permanently ice-covered lakes located in the McMurdo Dry Valleys, Antarctica. The perennial ice cover maintains a chemically stratified water column and unlike other inland bodies of water, largely prevents external input of carbon and nutrients from streams. Biota are exposed to numerous environmental stresses, including year-round severe nutrient deficiency, low temperatures, extreme shade, hypersalinity, and 24-hour darkness during the winter (1). These extreme environmental conditions limit the biota in Lake Bonney almost exclusively to microorganisms (2). Single-celled microbial eukaryotes (called "protists") are important players in global biogeochemical cycling (3) and play important ecological roles in the cycling of carbon in the dry valley lakes, occupying both primary and tertiary roles in the aquatic food web. In the dry valley aquatic food web, protists that fix inorganic carbon (autotrophy) are the major producers of organic carbon for organotrophic organisms (4, 2). Phagotrophic or heterotrophic protists capable of ingesting bacteria and smaller protists act as the top predators in the food web (5). Last, an unknown proportion of the protist population is capable of combined mixotrophic metabolism (6, 7). Mixotrophy in protists involves the ability to combine photosynthetic capability with phagotrophic ingestion of prey microorganisms. This form of mixotrophy differs from mixotrophic metabolism in bacterial species, which generally involves uptake dissolved carbon molecules. There are currently very few protist isolates from permanently ice-capped polar lakes, and studies of protist diversity and ecology in this extreme environment have been limited (8, 4, 9, 10, 5). A better understanding of protist metabolic versatility in the simple dry valley lake food web will aid in the development of models for the role of protists in the global carbon cycle. We employed an enrichment culture approach to isolate potentially

  4. Geochemical diversity in S processes mediated by culture-adapted and environmental-enrichments of Acidithiobacillus spp.

    Science.gov (United States)

    Bernier, Luc; Warren, Lesley A.

    2007-12-01

    Coupled S speciation and acid generation resulting from S processing associated with five different microbial treatments, all primarily Acidithiobacillus spp. (i.e. autotrophic S-oxidizers) were evaluated in batch laboratory experiments. Microbial treatments included two culture-adapted strains, Acidithiobacillus ferrooxidans and Acidithiobacillus thiooxidans, their consortia and two environmental enrichments from a mine tailings lake that were determined to be >95% Acidithiobacillus spp., by whole-cell fluorescent hybridization. Using batch experiments simulating acidic mine waters with no carbon amendments, acid generation, and S speciation associated with the oxidation of three S substrates (thiosulfate, tetrathionate, and elemental S) were evaluated. Aseptic controls showed no observable pH decrease over the experimental time course (1 month) for all three S compounds examined. In contrast, pH decreased in all microbial treatments from starting pH values of 4 to 2 or less for all three S substrates. Results show a non-linear relationship between the pH dynamics of the batch cultures and their corresponding sulfate concentrations, and indicate how known microbial S processing pathways have opposite impacts, ultimately on pH dynamics. Associated geochemical modeling indicated negligible abiogenic processes contributing to the observed results, indicating strong microbial control of acid generation extending over pH ranges from 4 to less than 2. However, the observed acid generation rates and associated S speciation were both microbial treatment and substrate-specific. Results reveal a number of novel insights regarding microbial catalysis of S oxidation: (1) metabolic diversity in S processing, as evidenced by the observed geochemical signatures in S chemical speciation and rates of acid generation amongst phylogenetically similar organisms (to the genus level); (2) consortial impacts differ from those of individual strain members; (3) environmental enrichments

  5. The metabolism of neonicotinoid insecticide thiamethoxam by soil enrichment cultures, and the bacterial diversity and plant growth-promoting properties of the cultured isolates.

    Science.gov (United States)

    Zhou, Guang-Can; Wang, Ying; Ma, Yuan; Zhai, Shan; Zhou, Ling-Yan; Dai, Yi-Jun; Yuan, Sheng

    2014-01-01

    A soil enrichment culture (SEC) rapidly degraded 96% of 200 mg L(-1) neonicotinoid insecticide thiamethoxam (TMX) in MSM broth within 30 d; therefore, its metabolic pathway of TMX, bacterial diversity and plant growth-promoting rhizobacteria (PGPR) activities of the cultured isolates were studied. The SEC transformed TMX via the nitro reduction pathway to form nitrso, urea metabolites and via cleavage of the oxadiazine cycle to form a new metabolite, hydroxyl CLO-tri. In addition, 16S rRNA gene-denaturing gradient gel electrophoresis analysis revealed that uncultured rhizobacteria are predominant in the SEC broth and that 77.8% of the identified bacteria belonged to uncultured bacteria. A total of 31 cultured bacterial strains including six genera (Achromobacter, Agromyces, Ensifer, Mesorhizobium, Microbacterium and Pseudoxanthomonas) were isolated from the SEC broth. The 12 strains of Ensifer adhaerens have the ability to degrade TMX. All six selected bacteria showed PGPR activities. E. adhaerens TMX-23 and Agromyces mediolanus TMX-25 produced indole-3-acetic acid, whereas E. adhaerens TMX-23 and Mesorhizobium alhagi TMX-36 are N2-fixing bacteria. The six-isolated microbes were tolerant to 200 mg L(-1) TMX, and the growth of E. adhaerens was significantly enhanced by TMX, whereas that of Achromobacter sp. TMX-5 and Microbacterium sp.TMX-6 were enhanced slightly. The present study will help to explain the fate of TMX in the environment and its microbial degradation mechanism, as well as to facilitate future investigations of the mechanism through which TMX enhances plant vigor.

  6. Biodegradation of Various Aromatic Compounds by Enriched Bacterial Cultures: Part B--Nitrogen-, Sulfur-, and Oxygen-Containing Heterocyclic Aromatic Compounds.

    Science.gov (United States)

    Oberoi, Akashdeep Singh; Philip, Ligy; Bhallamudi, S Murty

    2015-07-01

    Present study focused on the biodegradation of various heterocyclic nitrogen, sulfur, and oxygen (NSO) compounds using naphthalene-enriched culture. Target compounds in the study were pyridine, quinoline, benzothiophene, and benzofuran. Screening studies were carried out using different microbial consortia enriched with specific polycyclic aromatic hydrocarbon (PAH) and NSO compounds. Among different microbial consortia, naphthalene-enriched culture was the most efficient consortium based on high substrate degradation rate. Substrate degradation rate with naphthalene-enriched culture followed the order pyridine > quinoline > benzofuran > benzothiophene. Benzothiophene and benzofuran were found to be highly recalcitrant pollutants. Benzothiophene could not be biodegraded when concentration was above 50 mg/l. It was observed that 2-(1H)-quinolinone, benzothiophene-2-one, and benzofuran-2,3-dione were formed as metabolic intermediates during biodegradation of quinoline, benzothiophene, and benzofuran, respectively. Quinoline-N and pyridine-N were transformed into free ammonium ions during the biodegradation process. Biodegradation pathways for various NSO compounds are proposed. Monod inhibition model was able to simulate single substrate biodegradation kinetics satisfactorily. Benzothiophene and benzofuran biodegradation kinetics, in presence of acetone, was simulated using a generalized multi-substrate model.

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

    Science.gov (United States)

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

    2016-01-01

    Here we introduce a novel strategy for turbidimetric monitoring of bacterial growth in liquid culture. The instrumentation comprises a light source, a customized 3D printed culture tube holder and a miniaturized spectrophotometer, connected through optical cables. Due to its small footprint and the possibility to operate with external light, bacterial growth was directly monitored from culture tubes in a simple and versatile fashion. This new portable measurement technique was used to monitor the growth of facultative (Escherichia coli ATCC/25922, and Staphylococcus aureus ATCC/29213) and strictly (Butyrivibrio fibrisolvens JW11, Butyrivibrio proteoclasticus P18, and Propionibacterium acnes DSMZ 1897) anaerobic bacteria. For E. coli and S. aureus, the growth rates calculated from normalized optical density values were compared with those ones obtained using a benchtop spectrophotometer without significant differences (P = 0.256). For the strictly anaerobic species, a high precision (relative standard deviation < 3.5%) was observed between replicates up to 48 h. Regarding its potential for customization, this manifold could accommodate further developments for customized turbidimetric monitoring, such as the use of light-emitting diodes as a light source or flow cells. PMID:27630632

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

    Directory of Open Access Journals (Sweden)

    Margarida R. G. Maia

    2016-08-01

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

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

    Science.gov (United States)

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

    2016-01-01

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

  10. Detection of Mycoplasma ovipneumoniae and M. arginini in bighorn sheep using enrichment culture coupled with genus- and species-specific polymerase chain reaction.

    Science.gov (United States)

    Weiser, Glen C; Drew, Mark L; Cassirer, E Frances; Ward, Alton C S

    2012-04-01

    Mycoplasma species are of interest as possible primary pathogens in the pneumonia complex of bighorn sheep (Ovis canadensis). Previous investigations have not commonly detected low frequencies of Mycoplasma spp. from free-ranging bighorn sheep, possibly due to the fastidious and slow growth of these organisms. We developed a culture protocol that employed an average initial 3-day enrichment culture in liquid Hayflick broth in a CO(2)-enhanced atmosphere. The broth was plated to solid Hayflick medium and the cultures observed for growth for up to 30 days. Polymerase chain reaction (PCR) was performed on DNA isolated from the enrichment broth and on isolates obtained from culture using Mycoplasma genus-specific PCR assays and species-specific PCR assays for M. arginini and M. ovipneumoniae. Some cultures that grew on Hayflick plates were picked as single colonies but were mixed because two organisms may grow together and appear as a single colony. Culture and PCR tests produced similar results for M. arginini, but for M. ovipneumoniae, culture alone was less accurate than PCR. Use of genus-specific primers also may allow detection of other species in samples negative for M. arginini and M. ovipneumoniae. Two methods of transport from field to laboratory (Port-a-Cul™ tubes, cryoprotectant in liquid N(2) and Fisher Transport System) gave similar results under our study conditions.

  11. Anaerobic degradation of benzene by marine sulfate-reducing bacteria

    Science.gov (United States)

    Musat, Florin; Wilkes, Heinz; Musat, Niculina; Kuypers, Marcel; Widdel, Friedrich

    2010-05-01

    Benzene, the archetypal aromatic hydrocarbon is a common constituent of crude oil and oil-refined products. As such, it can enter the biosphere through natural oil seeps or as a consequence of exploitation of fossil fuel reservoirs. Benzene is chemically very stable, due to the stabilizing aromatic electron system and to the lack of functional groups. Although the anaerobic degradation of benzene has been reported under denitrifying, sulfate-reducing and methanogenic conditions, the microorganisms involved and the initial biochemical steps of degradation remain insufficiently understood. Using marine sediment from a Mediterranean lagoon a sulfate-reducing enrichment culture with benzene as the sole organic substrate was obtained. Application of 16S rRNA gene-based methods showed that the enrichment was dominated (more than 85% of total cells) by a distinct phylotype affiliated with a clade of Deltaproteobacteria that include degraders of other aromatic hydrocarbons, such as naphthalene, ethylbenzene and m-xylene. Using benzoate as a soluble substrate in agar dilution series, several pure cultures closely related to Desulfotignum spp. and Desulfosarcina spp. were isolated. None of these strains was able to utilize benzene as a substrate and hybridizations with specific oligonucleotide probes showed that they accounted for as much as 6% of the total cells. Incubations with 13C-labeled benzene followed by Halogen in situ Hybridization - Secondary Ion Mass Spectroscopy (HISH-SIMS) analysis showed that cells of the dominant phylotype were highly enriched in 13C, while the accompanying bacteria had little or no 13C incorporation. These results demonstrate that the dominant phylotype was indeed the apparent benzene degrader. Dense-cell suspensions of the enrichment culture did not show metabolic activity toward added phenol or toluene, suggesting that benzene degradation did not proceed through anaerobic hydroxylation or methylation. Instead, benzoate was identified in

  12. Comparison of polymerase chain reaction methods and plating for analysis of enriched cultures of Listeria monocytogenes when using the ISO11290-1 method.

    Science.gov (United States)

    Dalmasso, Marion; Bolocan, Andrei Sorin; Hernandez, Marta; Kapetanakou, Anastasia E; Kuchta, Tomáš; Manios, Stavros G; Melero, Beatriz; Minarovičová, Jana; Muhterem, Meryem; Nicolau, Anca Ioana; Rovira, Jordi; Skandamis, Panagiotis N; Stessl, Beatrix; Wagner, Martin; Jordan, Kieran; Rodríguez-Lázaro, David

    2014-03-01

    Analysis for Listeria monocytogenes by ISO11290-1 is time-consuming, entailing two enrichment steps and subsequent plating on agar plates, taking five days without isolate confirmation. The aim of this study was to determine if a polymerase chain reaction (PCR) assay could be used for analysis of the first and second enrichment broths, saving four or two days, respectively. In a comprehensive approach involving six European laboratories, PCR and traditional plating of both enrichment broths from the ISO11290-1 method were compared for the detection of L. monocytogenes in 872 food, raw material and processing environment samples from 13 different dairy and meat food chains. After the first and second enrichments, total DNA was extracted from the enriched cultures and analysed for the presence of L. monocytogenes DNA by PCR. DNA extraction by chaotropic solid-phase extraction (spin column-based silica) combined with real-time PCR (RTi-PCR) was required as it was shown that crude DNA extraction applying sonication lysis and boiling followed by traditional gel-based PCR resulted in fewer positive results than plating. The RTi-PCR results were compared to plating, as defined by the ISO11290-1 method. For first and second enrichments, 90% of the samples gave the same results by RTi-PCR and plating, whatever the RTi-PCR method used. For the samples that gave different results, plating was significantly more accurate for detection of positive samples than RTi-PCR from the first enrichment, but RTi-PCR detected a greater number of positive samples than plating from the second enrichment, regardless of the RTi-PCR method used. RTi-PCR was more accurate for non-food contact surface and food contact surface samples than for food and raw material samples especially from the first enrichment, probably because of sample matrix interference. Even though RTi-PCR analysis of the first enrichment showed less positive results than plating, in outbreak scenarios where a rapid result is

  13. In vitro activity of trovafloxacin against Bacteroides fragilis in mixed culture with either Escherichia coli or a vancomycin- resistant strain of Enterococcus faecium determined by an anaerobic time-kill technique.

    NARCIS (Netherlands)

    L.E.T. Stearne (Lorna); C. Kooi; W.H.F. Goessens (Wil); I.A.J.M. Bakker-Woudenberg (Irma); I.C. Gyssens (Inge)

    2001-01-01

    textabstractTo determine the efficacy of trovafloxacin as a possible treatment for intra-abdominal abscesses, we have developed an anaerobic time-kill technique using different inocula to study the in vitro killing of Bacteroides fragilis in pure culture or in mixed cul

  14. Legal Thinking on the Construction of Culture -Enriched Beijing%"人文北京"建设的法制思考

    Institute of Scientific and Technical Information of China (English)

    杨积堂

    2012-01-01

    The core of Culture - enriched Beijing construction lies in humanity and culture. From the perspec- tive of humanity, Culture - enriched Beijing construction aims to build up a more prosperous, civilized, harmoni- ous, livable and healthy socialist society. From the perspective of culture, Culture -enriched Beijing construction aims to build up the socialism with Chinese characteristics with the most humanistic concern, civilized and cultural charm capital. Legal construction is the significant aim and content requirement. For one thing, rule of law is one of content requirements to realize the real Culture - enriched Beijing. For another, perfecting the legal system is the fundamental guarantee in Culture -enriched Beijing construction. The perfect culture legal system, harmonious and efficient enforcement of cultural mechanism, uncorrupted judicial operation mechanism as well as convenient law intermediary service mechanism are the requirements of cultural legal construction. As for the legal system construc- tion, laying down the legal construction strategy, perfecting the local cultural code system and creating good legal atmosphere for Culture -enriched Beijing construction are proposed.%“人文北京”建设的核心是“人”和“文”,以人为本,以文化人,从“人”的角度,“人文北京”就是要建设更加繁荣、文明、和谐、宜居、健康的社会主义和谐社会“首善之区”。从“文”的角度.“人文北京”就是要建成最具人文关怀、最显文明风采、最有文化魅力的中国特色社会主义先进文化之都。“法治”建设,是人文北京建设的重要目标和内涵要求。一方面,法治是实现真正的“人文北京”的内涵之一;另一方面,健全法制又是“人文北京”建设的根本保障。完善的文化法律法规体系、协调高效的文化执法机制、廉洁公正的司法运行机制、便捷的法律中介服务机制是文化法制

  15. Preparation of glycerol-enriched yeast culture and its effect on blood metabolites and ruminal fermentation in goats.

    Directory of Open Access Journals (Sweden)

    Gengping Ye

    Full Text Available The aim of this study was to isolate a glycerol-producing yeast strain from nature to prepare glycerol-enriched yeast culture (GY, and preliminarily evaluate the effects of GY on blood metabolites and ruminal fermentation in goats. During the trial, six isolates were isolated from unprocessed honey, and only two isolates with higher glycerol yield were identified by analysis of 26S ribosomal DNA sequences. One of the two isolates was identified as Saccharomyces cerevisiae, a direct-fed microbe permitted by the FDA. This isolate was used to prepare GY. The fermentation parameters were optimized through single-factor and orthogonal design methods to maximize the glycerol yield and biomass. The final GY contained 38.7±0.6 g/L glycerol and 12.6±0.5 g/L biomass. In vivo, eight castrated male goats with ruminal fistula were used in a replicated 4×4 Latin square experiment with four consecutive periods of 15 d. Treatments were as follows: control, LGY, MGY, and HGY with 0, 100, 200, and 300 mL GY per goat per day, respectively. The GY was added in two equal portions at 08∶00 and 17∶00 through ruminal fistula. Samples of blood and ruminal fluid were collected on the last one and two days of each period, respectively. Results showed that the plasma concentrations of triglyceride and total cholesterol were not affected by the supplemented GY. Compared with the control, goats supplemented with MGY and HGY had significantly higher (P<0.05 concentrations of plasma glucose and total protein, ruminal volatile fatty acid and molar proportion of propionate, and significantly lower (P<0.05 ruminal pH and ammonia nitrogen. These parameters changed linearly with increasing GY supplementation level (P<0.05. In conclusion, GY has great potential to be developed as a feed additive with dual effects of glycerol and yeast for ruminants.

  16. Anaerobic bacteria

    Science.gov (United States)

    Brook I, Goldstein EJ. Diseases caused by non-spore forming anaerobic bacteria. In: Goldman L, Schafer AI, eds. Goldman's Cecil Medicine . 25th ed. Philadelphia, PA: Elsevier Saunders; 2015:chap 297. Stedman's Online ...

  17. Hydrogen and lipid production from starch wastewater by co-culture of anaerobic sludge and oleaginous microalgae with simultaneous COD, nitrogen and phosphorus removal.

    Science.gov (United States)

    Ren, Hong-Yu; Liu, Bing-Feng; Kong, Fanying; Zhao, Lei; Ren, Nanqi

    2015-11-15

    Anaerobic sludge (AS) and microalgae were co-cultured to enhance the energy conversion and nutrients removal from starch wastewater. Mixed ratio, starch concentration and initial pH played critical roles on the hydrogen and lipid production of the co-culture system. The maximum hydrogen production of 1508.3 mL L(-1) and total lipid concentration of 0.36 g L(-1) were obtained under the optimized mixed ratio (algae:AS) of 30:1, starch concentration of 6 g L(-1) and initial pH of 8. The main soluble metabolites in dark fermentation were acetate and butyrate, most of which can be consumed in co-cultivation. When sweet potato starch wastewater was used as the substrate, the highest COD, TN and TP removal and energy conversion efficiencies reached 80.5%, 88.7%, 80.1% and 34.2%, which were 176%, 178%, 200% and 119% higher than that of the control group (dark fermentation), respectively. This research provided a novel approach and achieved efficient simultaneous energy recovery and nutrients removal from starch wastewater by the co-culture system.

  18. 322份血培养阳性病原菌在需氧、厌氧瓶的检出率比较%Comparison of detection rates of 322 blood culture positive pathogens in aerobic and anaerobic bottles

    Institute of Scientific and Technical Information of China (English)

    陈玉莲; 徐涛; 李磊邦; 李景松; 李永赞

    2012-01-01

    OBJECTIVE To analyze the detection rates of blood culture-positive pathogens in the aerobic bottle and anaerobic bottle from Jul 2010 to Jul 2011. METHODS The culture and identification for the blood were performed with the United States BD 9050 automated blood culture system and French bio-Merieux ATBExpression automatic bacterial identification system. RESULTS Of 2700 blood culture specimens, 322 blood cultures were positive, the positive rate was 11. 9%, including gram-positive cocci (24. 5%), gram-positive bacilli (0. 6%), gram-negative cocci (0.3%), gram-negative bacilli (65.8%), anaerobic (2.0%), and fungi (6.8%); there were only 130 (40. 4%) positive blood cultures in aerobic bottle and 60 (18. 6%) positive blood cultures in anaerobic bottlei there were 132 (41. 0%} positive cultures in aerobic and anaerobic blood cultures. CONCLUSION Simultaneous culture with aerobic and anaerobic bottles for the same patient not only can fully reflect the rapidity of automatic blood culture system of rapid but also improve the positive rate of blood culture.%目的 分析2010年7月-2011年7月血培养阳性的病原菌在需氧、厌氧瓶检出率.方法 血培养及鉴定采用美国BD9050全自动血培养仪和法国生物梅里埃公司ATBExpression自动细菌鉴定仪.结果 在2700份血培养中阳性322份,阳性率为11.9%,其中革兰阳性球菌占24.5%、革兰阳性杆菌占0.6%、革兰阴性球菌占0.3%、革兰阴性杆菌占65.8%、厌氧菌占2.0%、真菌占6.8%;其中仅需氧血培养阳性130份,占40.4%,仅厌氧血培养阳性60份,占18.6%,需氧与厌氧血培养均阳性132份,占41.0%.结论 同一患者同时进行需氧瓶及厌氧瓶培养,即能充分体现自动血培养仪快速性,又能提高血培养阳性率.

  19. Phylogenetic and functional diversity within toluene-degrading, sulphate-reducing consortia enriched from a contaminated aquifer.

    Science.gov (United States)

    Kuppardt, Anke; Kleinsteuber, Sabine; Vogt, Carsten; Lüders, Tillmann; Harms, Hauke; Chatzinotas, Antonis

    2014-08-01

    Three toluene-degrading microbial consortia were enriched under sulphate-reducing conditions from different zones of a benzene, toluene, ethylbenzene and xylenes (BTEX) plume of two connected contaminated aquifers. Two cultures were obtained from a weakly contaminated zone of the lower aquifer, while one culture originated from the highly contaminated upper aquifer. We hypothesised that the different habitat characteristics are reflected by distinct degrader populations. Degradation of toluene with concomitant production of sulphide was demonstrated in laboratory microcosms and the enrichment cultures were phylogenetically characterised. The benzylsuccinate synthase alpha-subunit (bssA) marker gene, encoding the enzyme initiating anaerobic toluene degradation, was targeted to characterise the catabolic diversity within the enrichment cultures. It was shown that the hydrogeochemical parameters in the different zones of the plume determined the microbial composition of the enrichment cultures. Both enrichment cultures from the weakly contaminated zone were of a very similar composition, dominated by Deltaproteobacteria with the Desulfobulbaceae (a Desulfopila-related phylotype) as key players. Two different bssA sequence types were found, which were both affiliated to genes from sulphate-reducing Deltaproteobacteria. In contrast, the enrichment culture from the highly contaminated zone was dominated by Clostridia with a Desulfosporosinus-related phylotype as presumed key player. A distinct bssA sequence type with high similarity to other recently detected sequences from clostridial toluene degraders was dominant in this culture. This work contributes to our understanding of the niche partitioning between degrader populations in distinct compartments of BTEX-contaminated aquifers.

  20. Landfill Leachate as Enrichment Culture for Ammonia-Oxidizing Bacteria%利用垃圾渗滤液富集培养氨氧化菌

    Institute of Scientific and Technical Information of China (English)

    崔荣; 李金玲; 李凤德; 韩京龙

    2011-01-01

    Maintaining a certain amount of active ammonia-oxidizing bacteria (AOB) in activated sludge is essential for the biological nitrogen removal process. The addition of enriched AOB into activated sludge is an effective option to increase AOB population. To economically get enriched AOB and effectively treat landfill leachate, the feasibility of using landfill leachate as a culture for enriching AOB was examined. Leachate from the Yantai municipal landfill site was used as the culture, and returned sludge from the Xin' an River municipal sewage treatment plant A2/O process was used as seed sludge. The AOB were enriched by fed batch cultivation. The results showed that after four cultivation cycles, the population of cells in the enriched AOB was 5. 6 times higher than the original activated sludge. By adding 14.5% of enriched AOB after four cultivation cycles into the activated sludge, the ammonia oxidation rate increased by 65.4%, which confirmed that landfill leachate could be used as a culture for enriching AOB.%确保活性污泥中适当的氨氧化菌(AOB)的数量及活性对污水生物除氮过程至关重要,投加富集AOB是增加活性污泥中AOB浓度的方法之一.为了经济有效地获取富集的AOB并有效处理难降解的垃圾渗滤液,对利用垃圾渗滤液富集培养AOB的可行性进行了研究.采用烟台市生活垃圾填埋场的垃圾渗滤液作为培养基,利用辛安河污水处理厂A2/O工艺二沉池的回流污泥进行接种,通过更代方式富集培养AOB.结果显示:更代4次后,菌液中AOB的浓度增至原来的5.6倍;向活性污泥中投加14.5%的经过4次更代富集培养的AOB,氨氧化速率提高了65.4%,从而验证了利用垃圾渗滤液富集AOB是可行的.

  1. Anaerobic oxidation of methane in grassland soils used for cattle husbandry

    Directory of Open Access Journals (Sweden)

    A. Bannert

    2012-10-01

    Full Text Available While the importance of anaerobic methane oxidation has been reported for marine ecosystems, the role of this process in soils is still questionable. Grasslands used as pastures for cattle overwintering show an increase in anaerobic soil micro-sites caused by animal treading and excrement deposition. Therefore, anaerobic potential methane oxidation activity of severely impacted soil from a cattle winter pasture was investigated in an incubation experiment under anaerobic conditions using 13C-labelled methane. We were able to detect a high microbial activity utilizing CH4 as nutrient source shown by the respiration of 13CO2. Measurements of possible terminal electron acceptors for anaerobic oxidation of methane were carried out. Soil sulfate concentrations were too low to explain the oxidation of the amount of methane added, but enough nitrate and iron(III were detected. However, only nitrate was consumed during the experiment. 13C-PLFA analyses clearly showed the utilization of CH4 as nutrient source mainly by organisms harbouring 16:1ω7 PLFAs. These lipids were also found as most 13C-enriched fatty acids by Raghoebarsing et al. (2006 after addition of 13CH4 to an enrichment culture coupling denitrification of nitrate to anaerobic oxidation of methane. This might be an indication for anaerobic oxidation of methane by relatives of "Candidatus Methylomirabilis oxyfera" in the investigated grassland soil under the conditions of the incubation experiment.

  2. 海洋细菌增菌培养方法的研究%Cultural methods of enrichment in ocean bacteria

    Institute of Scientific and Technical Information of China (English)

    马聪; 蒋学兵; 刘敏; 张在文; 王海东; 陈昌国; 郭建巍

    2008-01-01

    Objective To explore the effecitve cultural methods of enrichment in ocean bacteria. Methods Different kinds of cultural mediums were selectd to culture seawater bacteria. Results Both alkaline peptone water( APW) and halophilic bacteria liquid had good effects to Vibrio enrichment. NO. 2 nutrient broth was effective for ordinary bacteria. Eumycete could be isolated from seawater by cultural sabouraud liquid. Conclusions APW and halophilic bacteria liquid should be selected for Vibrio enrichment, NO. 2 nutrient broth for ordinary bacteria. Moreover, cultural sabouraud liquid can be effective for fungus.%目的 探讨理想的海洋细菌增菌培养方法.方法 选择不同种类的增菌培养液对海水中细菌进行增菌培养.结果 碱性蛋白胨水与嗜盐菌增菌液对海水弧菌具有良好的增菌效果,2号营养肉汤适合海水普通细菌的增菌培养,液体沙保氏增菌液能从海水中增菌培养出真菌.结论 海水中弧菌增菌应选用碱性蛋白胨水与嗜盐菌增菌液,增菌普通细菌应选用2号营养肉汤,真菌应选用液体沙保氏增菌液.

  3. Transition of an Anaerobic Escherichia coli Culture to Aerobiosis: Balancing mRNA and Protein Levels in a Demand-Directed Dynamic Flux Balance Analysis.

    Directory of Open Access Journals (Sweden)

    Joachim von Wulffen

    Full Text Available The facultative anaerobic bacterium Escherichia coli is frequently forced to adapt to changing environmental conditions. One important determinant for metabolism is the availability of oxygen allowing a more efficient metabolism. Especially in large scale bioreactors, the distribution of oxygen is inhomogeneous and individual cells encounter frequent changes. This might contribute to observed yield losses during process upscaling. Short-term gene expression data exist of an anaerobic E. coli batch culture shifting to aerobic conditions. The data reveal temporary upregulation of genes that are less efficient in terms of energy conservation than the genes predicted by conventional flux balance analyses. In this study, we provide evidence for a positive correlation between metabolic fluxes and gene expression. We then hypothesize that the more efficient enzymes are limited by their low expression, restricting flux through their reactions. We define a demand that triggers expression of the demanded enzymes that we explicitly include in our model. With these features we propose a method, demand-directed dynamic flux balance analysis, dddFBA, bringing together elements of several previously published methods. The introduction of additional flux constraints proportional to gene expression provoke a temporary demand for less efficient enzymes, which is in agreement with the transient upregulation of these genes observed in the data. In the proposed approach, the applied objective function of growth rate maximization together with the introduced constraints triggers expression of metabolically less efficient genes. This finding is one possible explanation for the yield losses observed in large scale bacterial cultivations where steady oxygen supply cannot be warranted.

  4. Coexistence of a sulphate-reducing Desulfovibrio species and the dehalorespiring Desulfitobacterium frappieri TCE1 in defined chemostat cultures grown with various combinations of sulphate and tetrachloroethene

    NARCIS (Netherlands)

    Drzyzga, O; Gerritse, J; Dijk, JA; Elissen, H; Gottschal, JC

    2001-01-01

    A two-member co-culture consisting of the dehalorespiring Desulfitobacterium frappieri TCE1 and the sulphate-reducing Desulfovibrio sp. strain SULF1 was obtained via anaerobic enrichment from soil contaminated with tetrachloroethene (PCE), In this co-culture, PCE dechlorination to cis-dichloroethene

  5. Evaluation of the matrix effect of thermophilic anaerobic digestion on inactivation of infectious laryngotracheitis virus using real-time PCR and viral cell culture.

    Science.gov (United States)

    Gao, Tiejun; Bowlby, Evelyn; Tong, Yupin; Wu, John T Y; Wong, Lester; Tower, Robert J; Pang, Xiaoli; Li, Xiaomei

    2012-04-01

    The matrix effect of the thermophilic anaerobic digestion (TAD) process on inactivation of infectious laryngotracheitis virus (ILTV) was evaluated. Viral cell culture and real-time PCR were used for assessing removal of the viral infectivity and degradation of viral DNA, respectively. Results showed that the TAD-derived matrix alone can inactivate the virus and destroy the nucleic acid helix core of ILTV in a time-and- dose-dependent manner. No cytopathogenic effect (CPE) was observed in the cells exposed to ILTV pre-treated with TAD matrix for 1.5h in experiment 1 and for 16h in experiment 2. There was a significant statistical difference between TAD matrix treated and non-treated cultures (p<0.001, Chi-test). Amplifiable ILT viral DNA was reduced 2.27 log by 1.5h-treatment and was not present by 16h-treatment with TAD matrix, indicating complete viral DNA fragmentation. The TAD process is an environmentally friendly way for disposing of poultry biowaste and carcasses.

  6. Fermentative hydrogen production from xylose using anaerobic mixed microflora

    Energy Technology Data Exchange (ETDEWEB)

    Lin, Chiu-Yue; Cheng, Chao-Hui [BioHydrogen Laboratory, Department of Hydraulic Engineering, Feng Chia University, P.O. Box 25-123, Taichung 40724 (Taiwan)

    2006-06-15

    Sewage sludge microflora were anaerobically cultivated in a chemostat-type anaerobic bioreactor at a temperature of 35+/-1{sup o}C, pH of 7.1 and hydraulic retention time of 12h to determine the hydrogen production efficiency from xylose (20g-COD/L). This enriched microflora was used as a seed in batch experiments to investigate the pH and substrate concentration effects on hydrogen-producing bioactivity. It is demonstrated that the enriched mesophilic sewage sludge microflora with a continuous feeding can produce hydrogen from xylose with hydrogen content of 32% (v/v) in the biogas. Each mole of xylose yields 0.7moles of hydrogen and each gram of biomass produces 0.038moles of hydrogen per day. According to the batch test results, changes in pH and xylose concentration could enhance the microflora hydrogen production activity. Batch cultivation of this mixed microflora at pH values of 6-7 and xylose concentrations of 20g-COD/L resulted in high hydrogen production with a yield of 1.92-2.25mol-H{sub 2}/mol-xylose. This value is comparable to that from an enrichment culture. Strategies based on pH and xylose concentration controls for optimal hydrogen production from xylose using sewage sludge microflora are proposed. (author)

  7. Composting-Like Conditions Are More Efficient for Enrichment and Diversity of Organisms Containing Cellulase-Encoding Genes than Submerged Cultures.

    Science.gov (United States)

    Heiss-Blanquet, Senta; Fayolle-Guichard, Françoise; Lombard, Vincent; Hébert, Agnès; Coutinho, Pedro M; Groppi, Alexis; Barre, Aurélien; Henrissat, Bernard

    2016-01-01

    Cost-effective biofuel production from lignocellulosic biomass depends on efficient degradation of the plant cell wall. One of the major obstacles for the development of a cost-efficient process is the lack of resistance of currently used fungal enzymes to harsh conditions such as high temperature. Adapted, thermophilic microbial communities provide a huge reservoir of potentially interesting lignocellulose-degrading enzymes for improvement of the cellulose hydrolysis step. In order to identify such enzymes, a leaf and wood chip compost was enriched on a mixture of thermo-chemically pretreated wheat straw, poplar and Miscanthus under thermophile conditions, but in two different set-ups. Unexpectedly, metagenome sequencing revealed that incubation of the lignocellulosic substrate with compost as inoculum in a suspension culture resulted in an impoverishment of putative cellulase- and hemicellulase-encoding genes. However, mimicking composting conditions without liquid phase yielded a high number and diversity of glycoside hydrolase genes and an enrichment of genes encoding cellulose binding domains. These identified genes were most closely related to species from Actinobacteria, which seem to constitute important players of lignocellulose degradation under the applied conditions. The study highlights that subtle changes in an enrichment set-up can have an important impact on composition and functions of the microcosm. Composting-like conditions were found to be the most successful method for enrichment in species with high biomass degrading capacity.

  8. Effect of anaerobic blood culture on diagnosis and treatment of suspected bloodstream infections%厌氧血培养对疑似血流感染患者临床诊断治疗的影响

    Institute of Scientific and Technical Information of China (English)

    余世全; 周惠琴; 刘励军

    2012-01-01

    OBJECTIVE To study the effect of anaerobic blood culture on the positive rate of blood culture of the patients with suspected bloodstream infections so as to provide basis for the timely diagnosis and treatment. METHODS The blood sample were cultured by adopting two sets of aerobic and anaerobic blood culture bottles, the culture result was identified and analyzed by using VITEK-32 instrument and identification card after the detection with BACTEC-9120 full-automatic blood culture system. RESULTS Of 1744 blood culture samples submitted, there were 215 positive with the positive rate of 12. 33% , totally 80 strains of bacteria were isolated; there were 84 both aerobic and anaerobic culture positive, accounting for 39. 07%, and there were 111 aerobic culture positive with anaerobic culture negative, accounting for 51. 63%, and there were 20 aerobic culture negative with anaerobic culture positive, accounting for 9. 30% , as compared with the result of the aerobic and anaerobic blood cultures, the differences were statistically significant(^2 = 23. 536, P<0. 05); of the both aerobic and anaerobic blood culture positive samples, there were 5 strains of pathogens appeared as positive 2 days before the anaerobic blood culture, 2 strains as positive 1 day before the anaerobic blood culture. CONCLUSION The anaerobic blood culture can increase the positive rate of the blood culture of the patients with suspected bloodstream infections; two sets of aerobic and anaerobic blood culture mode can improve the positive rate of blood culture, ensure the acquirement of the culture result in the early stage, avoid the escaped examination, and diagnose and treat the diseases in a timely manner, which plays a significant role in improving the cure rate of the patients.%目的 探讨厌氧血培养对疑似血流感染患者血培养阳性率的影响,为临床及时正确的诊断及治疗提供依据.方法 血液标本采用双套需氧瓶加厌氧瓶血培养,用BACTEC-9120全自

  9. In-Situ Anaerobic Biosurfactant Production Process For Remediation Of DNAPL Contamination In Subsurface Aquifers

    Science.gov (United States)

    Albino, J. D.; Nambi, I. M.

    2009-12-01

    Microbial Enhanced Oil Recovery (MEOR) and remediation of aquifers contaminated with hydrophobic contaminants require insitu production of biosurfactants for mobilization of entrapped hydrophobic liquids. Most of the biosurfactant producing microorganisms produce them under aerobic condition and hence surfactant production is limited in subsurface condition due to lack of oxygen. Currently bioremediation involves expensive air sparging or excavation followed by exsitu biodegradation. Use of microorganisms which can produce biosurfactants under anaerobic conditions can cost effectively expedite the process of insitu bioremediation or mobilization. In this work, the feasibility of anaerobic biosurfactant production in three mixed anaerobic cultures prepared from groundwater and soil contaminated with chlorinated compounds and municipal sewage sludge was investigated. The cultures were previously enriched under complete anaerobic conditions in the presence of Tetrachloroethylene (PCE) for more than a year before they were studied for biosurfactant production. Biosurfactant production under anaerobic conditions was simulated using two methods: i) induction of starvation in the microbial cultures and ii) addition of complex fermentable substrates. Positive result for biosurfactant production was not observed when the cultures were induced with starvation by adding PCE as blobs which served as the only terminal electron acceptor. However, slight reduction in interfacial tension was noticed which was caused by the adherence of microbes to water-PCE interface. Biosurfactant production was observed in all the three cultures when they were fed with complex fermentable substrates and surface tension of the liquid medium was lowered below 35 mN/m. Among the fermentable substrates tested, vegetable oil yielded highest amount of biosurfactant in all the cultures. Complete biodegradation of PCE to ethylene at a faster rate was also observed when vegetable oil was amended to the

  10. Multiplexed Single Intact Cell Droplet Digital PCR (MuSIC ddPCR) Method for Specific Detection of Enterohemorrhagic E. coli (EHEC) in Food Enrichment Cultures

    Science.gov (United States)

    McMahon, Tanis C.; Blais, Burton W.; Wong, Alex; Carrillo, Catherine D.

    2017-01-01

    Foodborne illness attributed to enterohemorrhagic E. coli (EHEC), a highly pathogenic subset of Shiga toxin-producing E. coli (STEC), is increasingly recognized as a significant public health issue. Current microbiological methods for identification of EHEC in foods often use PCR-based approaches to screen enrichment broth cultures for characteristic gene markers [i.e., Shiga toxin (stx) and intimin (eae)]. However, false positives arise when complex food matrices, such as beef, contain mixtures of eae-negative STEC and eae-positive E. coli, but no EHEC with both markers in a single cell. To reduce false-positive detection of EHEC in food enrichment samples, a Multiplexed, Single Intact Cell droplet digital PCR (MuSIC ddPCR) assay capable of detecting the co-occurrence of the stx and eae genes in a single bacterial cell was developed. This method requires: (1) dispersal of intact bacteria into droplets; (2) release of genomic DNA (gDNA) by heat lysis; and (3) amplification and detection of genetic targets (stx and eae) using standard TaqMan chemistries with ddPCR. Performance of the method was tested with panels of EHEC and non-target E. coli. By determining the linkage (i.e., the proportion of droplets in which stx and eae targets were both amplified), samples containing EHEC (typically greater than 20% linkage) could be distinguished from samples containing mixtures of eae-negative STEC and eae-positive E. coli (0–2% linkage). The use of intact cells was necessary as this linkage was not observed with gDNA extracts. EHEC could be accurately identified in enrichment broth cultures containing excess amounts of background E. coli and in enrichment cultures derived from ground beef/pork and leafy-green produce samples. To our knowledge, this is the first report of dual-target detection in single bacterial cells using ddPCR. The application of MuSIC ddPCR to enrichment-culture screening would reduce false-positives, thereby improving the cost, speed, and accuracy of

  11. Enrichment of prostate cancer stem-like cells from human prostate cancer cell lines by culture in serum-free medium and chemoradiotherapy.

    Science.gov (United States)

    Wang, Lei; Huang, Xing; Zheng, Xinmin; Wang, Xinghuan; Li, Shiwen; Zhang, Lin; Yang, Zhonghua; Xia, Zhiping

    2013-01-01

    The discovery of rare subpopulations of cancer stem cells (CSCs) has created a new focus in cancer research. As CSCs demonstrate resistance to chemoradiation therapy relative to other cancer cells, this allows the enrichment of CSC populations by killing apoptosis-susceptible cancer cells. In this study, three commonly used human prostate cancer (PCa) cell lines (DU145, PC-3 and LNCaP) were examined for their expression of the putative stem cell markers CD133 and CD44 via flow cytometric analysis. Under normal culture conditions, CD133(+)/CD44(+) cells were only present in the DU145 cell line, and comprised only a minor percentage (0.1% ± 0.01%) of the total population. However, the proportion of these CD133(+)/CD44(+) prostate CSCs could be increased in these cell lines via culture in serum-free medium (SFM), or through chemotherapy or radiotherapy. Indeed, after culture in SFM, the proportion of CD133(+)/CD44(+) cells in DU145 and PC-3 had increased to 10.3% and 3.0%, respectively. Moreover, the proportion had increased to 9.8% enriched by chemotherapy and 3.5% by radiotherapy in DU145. Colony-formation tests, cell invasion assays, and tumor xenografts in BALB/c nude mice were used to evaluate the stem cell properties of CD133(+)/CD44(+) PCa cells that were isolated via fluorescence-activated cell sorting (FACS). CD133(+)/CD44(+) cells had an enhanced colony-formation capability and invasive ability in vitro, and displayed greater tumorigenic properties in vivo. These results demonstrate the presence of CD133(+)/CD44(+) prostate CSCs in established PCa cell lines and that populations of these cells can be enriched by culture in SFM or chemoradiotherapy. Finding novel therapies to override chemoradiation resistance in the prostate CSCs is the key to improve long-term results in PCa management.

  12. Quantification of enteric viruses, pathogen indicators, and Salmonella bacteria in class B anaerobically digested biosolids by culture and molecular methods.

    Science.gov (United States)

    Wong, Kelvin; Onan, Brandon M; Xagoraraki, Irene

    2010-10-01

    The most common class B biosolids in the United States are generated by mesophilic anaerobic digestion (MAD), and MAD biosolids have been used for land application. However, the pathogen levels in MAD biosolids are still unclear, especially with respect to enteric viruses. In this study, we determined the occurrence and the quantitative levels of enteric viruses and indicators in 12 MAD biosolid samples and of Salmonella enterica in 6 MAD biosolid samples. Three dewatered biosolid samples were also included in this study for purposes of comparison. Human adenoviruses (HAdV) had the highest gene levels and were detected more frequently than other enteric viruses. The gene levels of noroviruses (NV) reported were comparable to those of enteroviruses (EV) and human polyomaviruses (HPyV). The occurrence percentages of HAdV, HAdV species F, EV, NV GI, NV GII, and HPyV in MAD samples were 83, 83, 42, 50, 75, and 58%, respectively. No hepatitis A virus was detected. Infectious HAdV was detected more frequently than infectious EV, and all infectious HAdV were detected when samples were propagated in A549 cells. Based on most-probable-number (MPN) analysis, A549 cells were more susceptible to biosolid-associated viruses than BGM cells. All indicator levels in MAD biosolids were approximately 10(4) MPN or PFU per gram (dry), and the dewatered biosolids had significantly higher indicator levels than the MAD biosolids. Only two MAD samples tested positive for Salmonella enterica, where the concentration was below 1.0 MPN/4 g. This study provides a broad comparison of the prevalence of different enteric viruses in MAD biosolids and reports the first detection of noroviruses in class B biosolids. The observed high quantitative and infectivity levels of adenoviruses in MAD biosolids indicate that adenovirus is a good indicator for the evaluation of sludge treatment efficiency.

  13. 富铬蚁巢伞(鸡(土从)菌)液体培养%Submerged culture of chromium-enriched Termitomyces albuminosus

    Institute of Scientific and Technical Information of China (English)

    胡忠策; 郑晓冬; 陈新爱; 郑裕国

    2008-01-01

    A species of mushroom, Termitomyces albuminosus, was cultured in liquid medium for production of chromium-enriched mycelium. The influence of chromium (Ⅲ) on mycelial growth of T. albuminosus was investigated. An optimum medium composed of 5.6g/L yeast extract, 51.6g/L hydrolyzed rice, 2g/L KH2PO4,and 20mg/L chromium(Ⅲ) with initial pH of 4.5 was obtained by using method of central composite design (CCD). After incubation of 84h, the maximal biomass of chromium-enriched mycelia reached 24.23g DMW(dried mycelial weight)/L with 272μg/g DMW chromium content in 500mL flasks containing 100mL medium with an inoculum of 8% on a shaker of 100r/min under an optimized cultivation condition at 28℃.

  14. Validation of Performance of Plastic versus Glass Bottles for Culturing Anaerobes from Blood in BacT/ALERT SN Medium

    OpenAIRE

    Mirrett, Stanley; Joyce, Maria J.; Reller, L. Barth

    2005-01-01

    To validate performance, we compared the new plastic BacT/ALERT (bioMérieux, Durham, NC) SN bottle to the current glass SN bottle with samples of blood obtained for culture from adults and found them comparable for both recovery and speed of detection of microorganisms. We conclude that the safety advantage of plastic bottles can be achieved without compromising performance.

  15. Validation of performance of plastic versus glass bottles for culturing anaerobes from blood in BacT/ALERT SN medium.

    Science.gov (United States)

    Mirrett, Stanley; Joyce, Maria J; Reller, L Barth

    2005-12-01

    To validate performance, we compared the new plastic BacT/ALERT (bioMérieux, Durham, NC) SN bottle to the current glass SN bottle with samples of blood obtained for culture from adults and found them comparable for both recovery and speed of detection of microorganisms. We conclude that the safety advantage of plastic bottles can be achieved without compromising performance.

  16. Addition of novobiocin in pre-enrichment step can improve Salmonella culture protocol of modified semisolid Rappaport-Vassiliadis

    DEFF Research Database (Denmark)

    Jensen, Annette Nygaard; Sørensen, Gitte; Baggesen, Dorte Lau

    2003-01-01

    The aim was to investigate the effect of addition of Novobiocin to the non-selective buffered peptone water (BPW) for pre-enrichment of Salmonella in connection with plating on modified semisolid Rappaport-Vassiliadis (MSRV). In a semi-quantitative study, the level of Salmonella following pre-enr...

  17. Biosynthesis of highly enriched 13C-lycopene for human metabolic studies using repeated batch tomato cell culturing with 13C-glucose.

    Science.gov (United States)

    Moran, Nancy Engelmann; Rogers, Randy B; Lu, Chi-Hua; Conlon, Lauren E; Lila, Mary Ann; Clinton, Steven K; Erdman, John W

    2013-08-15

    While putative disease-preventing lycopene metabolites are found in both tomato (Solanum lycopersicum) products and in their consumers, mammalian lycopene metabolism is poorly understood. Advances in tomato cell culturing techniques offer an economical tool for generation of highly-enriched (13)C-lycopene for human bioavailability and metabolism studies. To enhance the (13)C-enrichment and yields of labelled lycopene from the hp-1 tomato cell line, cultures were first grown in (13)C-glucose media for three serial batches and produced increasing proportions of uniformly labelled lycopene (14.3±1.2%, 39.6±0.5%, and 48.9±1.5%) with consistent yields (from 5.8 to 9 mg/L). An optimised 9-day-long (13)C-loading and 18-day-long labelling strategy developed based on glucose utilisation and lycopene yields, yielded (13)C-lycopene with 93% (13)C isotopic purity, and 55% of isotopomers were uniformly labelled. Furthermore, an optimised acetone and hexane extraction led to a fourfold increase in lycopene recovery from cultures compared to a standard extraction.

  18. Epidemiology of Salmonella sp. in California cull dairy cattle: prevalence of fecal shedding and diagnostic accuracy of pooled enriched broth culture of fecal samples

    Directory of Open Access Journals (Sweden)

    Omran A. Abu Aboud

    2016-08-01

    Full Text Available Background The primary objective of this cross-sectional study was to estimate the crude, seasonal and cull-reason stratified prevalence of Salmonella fecal shedding in cull dairy cattle on seven California dairies. A secondary objective was to estimate and compare the relative sensitivity (Se and specificity (Sp for pools of 5 and 10 enriched broth cultures of fecal samples for Salmonella sp. detection. Methods Seven dairy farms located in the San Joaquin Valley of California were identified and enrolled in the study as a convenience sample. Cull cows were identified for fecal sampling once during each season between 2014 and 2015, specifically during spring, summer, fall, and winter, and 10 cows were randomly selected for fecal sampling at the day of their sale. In addition, study personnel completed a survey based on responses of the herd manager to questions related to the previous four month’s herd management. Fecal samples were frozen until testing for Salmonella. After overnight enrichment in liquid broth, pools of enrichment broth (EBP were created for 5 and 10 samples. All individual and pooled broths were cultured on selective media with putative Salmonella colonies confirmed by biochemical testing before being serogrouped and serotyped. Results A total of 249 cull cows were enrolled into the study and their fecal samples tested for Salmonella. The survey-weighted period prevalence of fecal shedding of all Salmonella sp. in the cull cow samples across all study herds and the entire study period was 3.42% (N = 249; SE 1.07. The within herd prevalence of Salmonella shed in feces did not differ over the four study seasons (P = 0.074. The Se of culture of EBP of five samples was 62.5% (SE = 17.12, which was not statistically different from the Se of culture of EBP of 10 (37.5%, SE = 17.12, P = 0.48. The Sp of culture of EBP of five samples was 95.24% (SE = 3.29 and for pools of 10 samples was 100.00% (SE = 0. There was no statistical

  19. Enrichment of Secondary Wastewater Sludge for Production of Hydrogen from Crude Glycerol and Comparative Evaluation of Mono-, Co- and Mixed-Culture Systems

    Directory of Open Access Journals (Sweden)

    Vinayak Laxman Pachapur

    2016-01-01

    Full Text Available Anaerobic digestion using mixed-culture with broader choice of pretreatments for hydrogen (H2 production was investigated. Pretreatment of wastewater sludge by five methods, such as heat, acid, base, microwave and chloroform was conducted using crude glycerol (CG as substrate. Results for heat treatment (100 °C for 15 min showed the highest H2 production across the pretreatment methods with 15.18 ± 0.26 mmol/L of medium at 30 °C in absence of complex media and nutrient solution. The heat-pretreated inoculum eliminated H2 consuming bacteria and produced twice as much as H2 as compared to other pretreatment methods. The fermentation conditions, such as CG concentration (1.23 to 24 g/L, percentage of inoculum size (InS (1.23% to 24% v/v along with initial pH (2.98 to 8.02 was tested using central composite design (CCD with H2 production as response parameter. The maximum H2 production of 29.43 ± 0.71 mmol/L obtained at optimum conditions of 20 g/L CG, 20% InS and pH 7. Symbiotic correlation of pH over CG and InS had a significant (p-value: 0.0011 contribution to H2 production. The mixed-culture possessed better natural acclimatization activity for degrading CG, at substrate inhibition concentration and provided efficient inoculum conditions in comparison to mono- and co-culture systems. The heat pretreatment step used across mixed-culture system is simple, cheap and industrially applicable in comparison to mono-/co-culture systems for H2 production.

  20. Enrichment of Secondary Wastewater Sludge for Production of Hydrogen from Crude Glycerol and Comparative Evaluation of Mono-, Co- and Mixed-Culture Systems.

    Science.gov (United States)

    Pachapur, Vinayak Laxman; Kutty, Prianka; Brar, Satinder Kaur; Ramirez, Antonio Avalos

    2016-01-13

    Anaerobic digestion using mixed-culture with broader choice of pretreatments for hydrogen (H₂) production was investigated. Pretreatment of wastewater sludge by five methods, such as heat, acid, base, microwave and chloroform was conducted using crude glycerol (CG) as substrate. Results for heat treatment (100 °C for 15 min) showed the highest H₂ production across the pretreatment methods with 15.18 ± 0.26 mmol/L of medium at 30 °C in absence of complex media and nutrient solution. The heat-pretreated inoculum eliminated H₂ consuming bacteria and produced twice as much as H₂ as compared to other pretreatment methods. The fermentation conditions, such as CG concentration (1.23 to 24 g/L), percentage of inoculum size (InS) (1.23% to 24% v/v) along with initial pH (2.98 to 8.02) was tested using central composite design (CCD) with H₂ production as response parameter. The maximum H₂ production of 29.43 ± 0.71 mmol/L obtained at optimum conditions of 20 g/L CG, 20% InS and pH 7. Symbiotic correlation of pH over CG and InS had a significant (p-value: 0.0011) contribution to H₂ production. The mixed-culture possessed better natural acclimatization activity for degrading CG, at substrate inhibition concentration and provided efficient inoculum conditions in comparison to mono- and co-culture systems. The heat pretreatment step used across mixed-culture system is simple, cheap and industrially applicable in comparison to mono-/co-culture systems for H₂ production.

  1. Non-cultured adipose-derived CD45(-) side population cells are enriched for progenitors that give rise to myofibres in vivo

    DEFF Research Database (Denmark)

    Andersen, Ditte C; Schrøder, Henrik D; Jensen, Charlotte H

    2008-01-01

    skeletal muscle repair mainly relies on the satellitecell, several reports have shown that vessel-associated cells may adopt a myogenic phenotype when exposed to a muscle environment. In accordance with these findings, we also observed invitro myogenic specification of SPCD45(-) cells when cocultured...... with myoblasts. Furthermore, immediate intramuscular engraftment of non-cultured SPCD45(-) cells gave rise to myofibres andcells lining blood vessels, whereas the SVF only provided donor derived mononuclear cells. We therefore conclude that the SPCD45(-) fraction of adipose-derived SVF is enriched for cells...

  2. Fermentation and growth response of a primary poultry isolate of Salmonella typhimurium grown under strict anaerobic conditions in continuous culture and amino acid-limited batch culture.

    Science.gov (United States)

    Maciorowski, K G; Nisbet, D J; Ha, S D; Corrier, D E; DeLoach, J R; Ricke, S C

    1997-01-01

    Salmonella typhimurium is a significant hazard to consumer health that is carried asymptomatically in poultry gastrointestinal tracts. Nurmi cultures may prevent Salmonella colonization in young chicks, but the mechanism of competitive exclusion is unclear. Modeling Salmonella's metabolism in pure culture may allow for greater definition in choosing strains for Nurmi cultures. The growth rates and affinity constants of S. typhimurium growing in amino acid-limited conditions were determined in batch culture and compared to primary poultry isolates of cecal strains. Serine and NH4Cl were the best N sources for growth of all organisms tested in this study. The fermentation response of S. typhimurium was also monitored in continuous culture at a slow dilution rate of 0.021 h-1. S. typhimurium was found to adapt to VL media, with trends in protein disappearance, Yglucose, and Yprotein. This may show that amino acid or protein concentrations may be an integral component of the initial establishment of S. typhimurium in the cecum.

  3. Metabolic interactions between methanogenic consortia and anaerobic respiring bacteria

    DEFF Research Database (Denmark)

    Stams, A.J.; Oude Elferink, S.J.; Westermann, Peter

    2003-01-01

    Most types of anaerobic respiration are able to outcompete methanogenic consortia for common substrates if the respective electron acceptors are present in sufficient amounts. Furthermore, several products or intermediate compounds formed by anaerobic respiring bacteria are toxic to methanogenic...... consortia. Despite the potentially adverse effects, only few inorganic electron acceptors potentially utilizable for anaerobic respiration have been investigated with respect to negative interactions in anaerobic digesters. In this chapter we review competitive and inhibitory interactions between anaerobic...... respiring populations and methanogenic consortia in bioreactors. Due to the few studies in anaerobic digesters, many of our discussions are based upon studies of defined cultures or natural ecosystems...

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

    DEFF Research Database (Denmark)

    Justesen, Tage; Justesen, Ulrik Stenz

    2013-01-01

    The maintenance of a strict anaerobic atmosphere is essential for the culture of strict anaerobic bacteria. We describe a simple and sensitive quality control method of the anaerobic atmosphere, based on the measurement of the zone diameter around a 5-μg metronidazole disk when testing an aerotol...

  5. Anaerobic biodegradability of macropollutants

    DEFF Research Database (Denmark)

    Angelidaki, Irini

    2002-01-01

    A variety of test procedures for determination of anaerobic biodegradability has been reported. This paper reviews the methods developed for determination of anaerobic biodegradability of macro-pollutants. Anaerobic biodegradability of micro-pollutants is not included. Furthermore, factors import...

  6. Non-Neuronal Cells Are Required to Mediate the Effects of Neuroinflammation: Results from a Neuron-Enriched Culture System.

    Directory of Open Access Journals (Sweden)

    Chin Wai Hui

    Full Text Available Chronic inflammation is associated with activated microglia and reactive astrocytes and plays an important role in the pathogenesis of neurodegenerative diseases such as Alzheimer's. Both in vivo and in vitro studies have demonstrated that inflammatory cytokine responses to immune challenges contribute to neuronal death during neurodegeneration. In order to investigate the role of glial cells in this phenomenon, we developed a modified method to remove the non-neuronal cells in primary cultures of E16.5 mouse cortex. We modified previously reported methods as we found that a brief treatment with the thymidine analog, 5-fluorodeoxyuridine (FdU, is sufficient to substantially deplete dividing non-neuronal cells in primary cultures. Cell cycle and glial markers confirm the loss of ~99% of all microglia, astrocytes and oligodendrocyte precursor cells (OPCs. More importantly, under this milder treatment, the neurons suffered neither cell loss nor any morphological defects up to 2.5 weeks later; both pre- and post-synaptic markers were retained. Further, neurons in FdU-treated cultures remained responsive to excitotoxicity induced by glutamate application. The immunobiology of the FdU culture, however, was significantly changed. Compared with mixed culture, the protein levels of NFκB p65 and the gene expression of several cytokine receptors were altered. Individual cytokines or conditioned medium from β-amyloid-stimulated THP-1 cells that were, potent neurotoxins in normal, mixed cultures, were virtually inactive in the absence of glial cells. The results highlight the importance of our glial-depleted culture system and identifies and offer unexpected insights into the complexity of -brain neuroinflammation.

  7. 血培养需氧及厌氧瓶阳性检出率与报警时间分析%Analysis of positive rate and alarm time of aerobic and anaerobic blood culture bottle

    Institute of Scientific and Technical Information of China (English)

    答嵘; 王伟; 马晨; 俱西驰

    2015-01-01

    OBJECTIVE To analyze the positive rate ,pathogenic distribution and alarm time of aerobic and anaero‐bic bottles in blood culture so as to provide clinical references for treatments .METHODS Totally 1684 sets of blood culture bottle were collected from Sep .to Nov .2013 ,and they were cultivated by BacT/ALERT 3D auto‐matic bacteria/mycobacterial culture .Identification and drug sensitive test were performed by VITEK‐2 Compact automatic microbe cultivation and drug susceptibility instrument . The data were analyzed by SPSS 18 .0 . RESULTS In this study ,1684 sets of blood culture bottle including aerobic and anaerobic bottles were collected . Totally 1498 sets were double negative ,and 101 sets were double positive .Generally 66 sets of bottle were aerobic positive and anaerobic negative ,while 19 sets of bottle were anaerobic positive and aerobic negative .Positive rate of aerobic bottle was 9 .92% ,while that of anaerobic bottles was 7 .13% .The detection rate of Acinetobacter , Brucella ,Micrococcus ,Corynebacterium and fungi in aerobic bottle was 100% ,while anaerobic bacteria in anae‐robic bottle was 100% .Alarm time of aerobic and anaerobic bottles had no statistical differences .CONCLUSION Blood culture of double sets of aerobic and anaerobic blood culture bottles is recommended ,which improves the positive rate and discriminate pollution .%目的:分析成套血培养中需氧与厌氧瓶的阳性检出率、病原菌分布与报警时间的特点,为临床诊断提供参考依据。方法收集2013年9-11月血培养瓶1684套,采用法国生物梅里埃公司BacT/ALERT 3D全自动细菌/分枝杆菌培养仪进行培养,阳性报警时转种采用VIT EK‐2 Compact全自动微生物鉴定与药敏仪进行病原菌鉴定与药敏试验,采用SPSS 18.0软件进行数据统计分析。结果收集的1684套血培养瓶中需氧瓶与厌氧瓶同时阴性1498套,同时阳性101套,需氧瓶阳性而厌氧瓶阴性66套,需

  8. Bacterial Wound Culture

    Science.gov (United States)

    ... Home Visit Global Sites Search Help? Bacterial Wound Culture Share this page: Was this page helpful? Also known as: Aerobic Wound Culture; Anaerobic Wound Culture Formal name: Culture, wound Related ...

  9. Comparative analysis of tertiary alcohol esterase activity in bacterial strains isolated from enrichment cultures and from screening strain libraries.

    Science.gov (United States)

    Herter, Susanne; Nguyen, Giang-Son; Thompson, Mark L; Steffen-Munsberg, Fabian; Schauer, Frieder; Bornscheuer, Uwe T; Kourist, Robert

    2011-05-01

    The preparation of enantiopure tertiary alcohols is of great contemporary interest due to the application of these versatile building blocks in organic synthesis and as precursors towards high value pharmaceutical compounds. Herein, we describe two approaches taken towards the discovery of novel biocatalysts for the synthesis of these valuable compounds. The first approach was initiated with screening of 47 bacterial strains for hydrolytic activity towards the simple tertiary alcohol ester tert-butyl acetate. In conjunction, a second method focussed on the isolation of strains competent for growth on tert-butyl acetate as the sole source of carbon and energy. From functional screening, 10 Gram-positive Actinomycetes showed hydrolytic activity, whilst enrichment selection resulted in the identification of 14 active strains, of which five belong to the Gram-negative cell-wall type. Bacterial strains obtained from both approaches were viable for enantioselective hydrolysis of pyridine substituted tertiary alcohol esters in addition to bulky aliphatic and keto-derived substrates from the same class. Activity towards each of the test substrates was uncovered, with promising enantioselectivities of up to E = 71 in the hydrolysis of a para-substituted pyridine tertiary alcohol ester using a strain of Rhodococcus ruber. Interestingly strains of Microbacterium and Alcaligenes sp. gave opposite enantiopreference in the hydrolysis of a meta-substituted pyridine tertiary alcohol ester with E values of 17 and 54. These approaches show that via both possibilities, screening established strain collections and performing enrichment selection, it is possible to identify novel species which show activity towards sterically challenging substrates.

  10. ENRICHMENT AND CHARACTERIZATION OF THYMUS-REPOPULATING CELLS IN STROMA-DEPENDENT CULTURES OF RAT BONE-MARROW

    NARCIS (Netherlands)

    PRAKAPAS, Z; DENOYELLE, M; DARGEMONT, C; KROESE, FGM; THIERY, JP; DEUGNIER, MA

    1993-01-01

    The bone marrow precursor cells seeding the thymus have been difficult to investigate using fresh bone marrow and in vivo thymus reconstitution assays. We have therefore designed a short-term bone marrow culture system allowing the study of thymus-repopulating cells in the marrow microenvironment. L

  11. An enrichment and acclimation procedure to obtain photo heterotrophic cultures for H{sub 2} production from organic effluents

    Energy Technology Data Exchange (ETDEWEB)

    Acevedo-Benitez, J. A.; Ponce-Noyola, M. T.; Poggi-Varaldo, H. M.

    2009-07-01

    Production of H{sub 2} via photo heterotrophic is an attractive alternative, due to the capacity of photo heterotrophic organisms to convert the organic matter of effluents into H{sub 2}. The objective of our work was to develop a protocol for selecting undefined mixed cultures of photo heterotrophic microorganism with the capability of producing of H{sub 2}. (Author)

  12. Bringing the Past to the Present. : The use of tagging and storytelling for the enrichment of cultural heritage.

    NARCIS (Netherlands)

    Vliet, H.M.M. (Harry) van; Hekman, Erik

    2011-01-01

    The Internet’s dominant role in recent years has caused a change in the relationship between media producers, suppliers and consumers in the traditional media landscape. The cultural sector must therefore decide what to do with today’s digital media in response to the general public’s changing role,

  13. Anaerobic Thermophiles

    Directory of Open Access Journals (Sweden)

    Francesco Canganella

    2014-02-01

    Full Text Available 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

  14. Enhanced start-up of anaerobic facultatively autotrophic biocathodes in bioelectrochemical systems.

    Science.gov (United States)

    Zaybak, Zehra; Pisciotta, John M; Tokash, Justin C; Logan, Bruce E

    2013-12-01

    Biocathodes in bioelectrochemical systems (BESs) can be used to convert CO2 into diverse organic compounds through a process called microbial electrosynthesis. Unfortunately, start-up of anaerobic biocathodes in BESs is a difficult and time consuming process. Here, a pre-enrichment method was developed to improve start-up of anaerobic facultatively autotrophic biocathodes capable of using cathodes as the electron donor (electrotrophs) and CO2 as the electron acceptor. Anaerobic enrichment of bacteria from freshwater bog sediment samples was first performed in batch cultures fed with glucose and then used to inoculate BES cathode chambers set at -0.4V (versus a standard hydrogen electrode; SHE). After two weeks of heterotrophic operation of BESs, CO2 was provided as the sole electron acceptor and carbon source. Consumption of electrons from cathodes increased gradually and was sustained for about two months in concert with a significant decrease in cathode chamber headspace CO2. The maximum current density consumed was -34 ± 4 mA/m(2). Biosynthesis resulted in organic compounds that included butanol, ethanol, acetate, propionate, butyrate, and hydrogen gas. Bacterial community analyses based on 16S rRNA gene clone libraries revealed Trichococcus palustris DSM 9172 (99% sequence identity) as the prevailing species in biocathode communities, followed by Oscillibacter sp. and Clostridium sp. Isolates from autotrophic cultivation were most closely related to Clostridium propionicum (99% sequence identity; ZZ16), Clostridium celerecrescens (98-99%; ZZ22, ZZ23), Desulfotomaculum sp. (97%; ZZ21), and Tissierella sp. (98%; ZZ25). This pre-enrichment procedure enables simplified start-up of anaerobic biocathodes for applications such as electrofuel production by facultatively autotrophic electrotrophs.

  15. Enhanced start-up of anaerobic facultatively autotrophic biocathodes in bioelectrochemical systems

    KAUST Repository

    Zaybak, Zehra

    2013-12-01

    Biocathodes in bioelectrochemical systems (BESs) can be used to convert CO2 into diverse organic compounds through a process called microbial electrosynthesis. Unfortunately, start-up of anaerobic biocathodes in BESs is a difficult and time consuming process. Here, a pre-enrichment method was developed to improve start-up of anaerobic facultatively autotrophic biocathodes capable of using cathodes as the electron donor (electrotrophs) and CO2 as the electron acceptor. Anaerobic enrichment of bacteria from freshwater bog sediment samples was first performed in batch cultures fed with glucose and then used to inoculate BES cathode chambers set at -0.4V (versus a standard hydrogen electrode; SHE). After two weeks of heterotrophic operation of BESs, CO2 was provided as the sole electron acceptor and carbon source. Consumption of electrons from cathodes increased gradually and was sustained for about two months in concert with a significant decrease in cathode chamber headspace CO2. The maximum current density consumed was -34±4mA/m2. Biosynthesis resulted in organic compounds that included butanol, ethanol, acetate, propionate, butyrate, and hydrogen gas. Bacterial community analyses based on 16S rRNA gene clone libraries revealed Trichococcus palustris DSM 9172 (99% sequence identity) as the prevailing species in biocathode communities, followed by Oscillibacter sp. and Clostridium sp. Isolates from autotrophic cultivation were most closely related to Clostridium propionicum (99% sequence identity; ZZ16), Clostridium celerecrescens (98-99%; ZZ22, ZZ23), Desulfotomaculum sp. (97%; ZZ21), and Tissierella sp. (98%; ZZ25). This pre-enrichment procedure enables simplified start-up of anaerobic biocathodes for applications such as electrofuel production by facultatively autotrophic electrotrophs. © 2013 Elsevier B.V.

  16. Bacterial diversity of autotrophic enriched cultures from remote, glacial Antarctic, Alpine and Andean aerosol, snow and soil samples

    Directory of Open Access Journals (Sweden)

    E. González-Toril

    2009-01-01

    Full Text Available Four different communities and one culture of autotrophic microbial assemblages were obtained by incubation of samples collected from high elevation snow in the Alps (Mt. Blanc area and the Andes (Nevado Illimani summit, Bolivia, from Antarctic aerosol (French station Dumont d'Urville and a maritime Antarctic soil (King George Island, South Shetlands, Uruguay Station Artigas, in a minimal mineral (oligotrophic media. Molecular analysis of more than 200 16S rRNA gene sequences showed that all cultured cells belong to the Bacteria domain. Phylogenetic comparison with the currently available rDNA database allowed sequences belonging to Proteobacteria Alpha-, Beta- and Gamma-proteobacteria, Actinobacteria and Bacteroidetes phyla to be identified. The Andes snow culture was the richest in bacterial diversity (eight microorganisms identified and the marine Antarctic soil the poorest (only one. Snow samples from Col du Midi (Alps and the Andes shared the highest number of identified microorganisms (Agrobacterium, Limnobacter, Aquiflexus and two uncultured Alphaproteobacteria clones. These two sampling sites also shared four sequences with the Antarctic aerosol sample (Limnobacter, Pseudonocardia and an uncultured Alphaproteobacteriaclone. The only microorganism identified in the Antarctica soil (Brevundimonas sp. was also detected in the Antarctic aerosol. Most of the identified microorganisms had been detected previously in cold environments, marine sediments soils and rocks. Air current dispersal is the best model to explain the presence of very specific microorganisms, like those identified in this work, in environments very distant and very different from each other.

  17. Timeline of bio-hydrogen production by anaerobic digestion of biomass

    Directory of Open Access Journals (Sweden)

    Bernadette E. TELEKY

    2015-12-01

    Full Text Available Anaerobic digestion of biomass is a process capable to produce biohydrogen, a clean source of alternative energy. Lignocellulosic biomass from agricultural waste is considered a renewable energy source; therefore its utilization also contributes to the reduction of water, soil and air pollution. The study consists in five consecutive experiments designed to utilize anaerobic bacterial enrichment cultures originating from the Hungarian Lake, Hévíz. Wheat straw was used as complex substrate to produce hydrogen. The timeline evolution of hydrogen production was analyzed and modelled by two functions: Logistic and Boltzmann. The results proved that hydrogen production is significant, with a maximum of 0.24 mlN/ml and the highest hydrogen production occurs between the days 4-10 of the experiment.

  18. Heavy metal incorporation in foraminiferal calcite: results from multi-element enrichment culture experiments with Ammonia tepida

    Directory of Open Access Journals (Sweden)

    G.-J. Reichart

    2010-08-01

    Full Text Available The incorporation of heavy metals into carbonate tests of the shallow water benthic foraminifer Ammonia tepida was investigated under controlled laboratory conditions. Temperature, salinity, and pH of the culture solutions were kept constant throughout the duration of this experiment, while trace metal concentrations were varied. Concentrations of Ni, Cu, and Mn were set 5-, 10-, and 20 times higher than levels found in natural North Sea water; for reference, a control experiment with pure filtered natural North Sea water was also analysed. The concentrations of Cu and Ni from newly grown chambers were determined by means of both μ-synchrotron XRF and Laser Ablation Inductively Coupled Plasma Mass Spectroscopy (LA-ICP-MS. The results of both independent analytical techniques agreed within the analytical uncertainty. In general, the concentration of the analysed elements in the tests increased in line with their concentration in the culture solutions. Potential toxic and/or chemical competition effects might have resulted in the decreased incorporation of Ni and Cu into the calcite of the specimens exposed to the highest elemental concentrations. Mn incorporation exhibited large variability in the experiment with the 20-fold increased element concentrations, potentially due to antagonistic effects with Cu. The partition coefficients of Cu and Ni were calculated to be 0.14 ± 0.02 and 1.0 ± 0.5, respectively, whereas the partition coefficient of Mn was estimated to be least 2.4. These partition coefficients now open the way for reconstructing past concentrations for these elements in sea water.

  19. Mechanisms, Chemistry, and Kinetics of Anaerobic Biodegradation of cis-Dichloroethene and Vinyl Chloride

    Energy Technology Data Exchange (ETDEWEB)

    McCarty, P.L.; Spormann, A.M.

    2000-12-01

    Anaerobic biological processes can result in PCE and TCE destruction through conversion to cis-dichloroethene (cDCE) then to vinyl chloride (VC), and finally to ethene. Here, the chlorinated aliphatic hydrocarbons (CAHs) serve as electron acceptors in energy metabolism, requiring electron donors such as hydrogen from an external source. The purpose of this study was to learn more about the biochemistry of cDCE and VC conversion to ethene, to better understand the requirements for electron donors, and to determine factors affecting the rates of CAH degradation and organism growth. The biochemistry of reductive dehalogenation of VC was studied with an anaerobic mixed culture enriched on VC. In other studies on electron donor needs for dehalogenation of cDCE and VC, competition for hydrogen was found to occur between the dehalogenators and other microorganisms such as methanogens and homoacetogens in a benzoate-acclimated dehalogenating methanogenic mixed culture. Factors affecting the relative rates of destruction of the solvents and their intermediate products were evaluated. Studies using a mixed PCE-dehalogenating culture as well as the VC enrichment for biochemical studies suggested that the same species was involved in both cDCE and VC dechlorination, and that cDCE and VC competitively inhibited each other's dechlorination rate.

  20. Anaerobic benzene degradation by Gram-positive sulfate-reducing bacteria.

    Science.gov (United States)

    Abu Laban, Nidal; Selesi, Drazenka; Jobelius, Carsten; Meckenstock, Rainer U

    2009-06-01

    Despite its high chemical stability, benzene is known to be biodegradable with various electron acceptors under anaerobic conditions. However, our understanding of the initial activation reaction and the responsible prokaryotes is limited. In the present study, we enriched a bacterial culture that oxidizes benzene to carbon dioxide under sulfate-reducing conditions. Community analysis using terminal restriction fragment length polymorphism, 16S rRNA gene sequencing and FISH revealed 95% dominance of one phylotype that is affiliated to the Gram-positive bacterial genus Pelotomaculum showing that sulfate-reducing Gram-positive bacteria are involved in anaerobic benzene degradation. In order to get indications of the initial activation mechanism, we tested the substrate utilization, performed cometabolism tests and screened for putative metabolites. Phenol, toluene, and benzoate could not be utilized as alternative carbon sources by the benzene-degrading culture. Cometabolic degradation experiments resulted in retarded rates of benzene degradation in the presence of phenol whereas toluene had no effect on benzene metabolism. Phenol, 2-hydroxybenzoate, 4-hydroxybenzoate, and benzoate were identified as putative metabolites in the enrichment culture. However, hydroxylated aromatics were shown to be formed abiotically. Thus, the finding of benzoate as an intermediate compound supports a direct carboxylation of benzene as the initial activation mechanism but additional reactions leading to its formation cannot be excluded definitely.

  1. Anaerobic mineralization of 2,4,6-tribromophenol to CO2 by a synthetic microbial community comprising Clostridium, Dehalobacter, and Desulfatiglans.

    Science.gov (United States)

    Li, Zhiling; Yoshida, Naoko; Wang, Aijie; Nan, Jun; Liang, Bin; Zhang, Chunfang; Zhang, Dongdong; Suzuki, Daisuke; Zhou, Xue; Xiao, Zhixing; Katayama, Arata

    2015-01-01

    Anaerobic mineralization of 2,4,6-tribromophenol (2,4,6-TBP) was achieved by a synthetic anaerobe community comprising a highly enriched culture of Dehalobacter sp. phylotype FTH1 acting as a reductive debrominator; Clostridium sp. strain Ma13 acting as a hydrogen supplier via glucose fermentation; and a novel 4-chlorophenol-degrading anaerobe, Desulfatiglans parachlorophenolica strain DS. 2,4,6-TBP was debrominated to phenol by the combined action of Ma13 and FTH1, then mineralized into CO2 by sequential introduction of DS, confirmed using [ring-(14)C(U)] phenol. The optimum concentrations of glucose, SO4(2-), and inoculum densities were 0.5 or 2.5mM, 1.0 or 2.5mM, and the densities equivalent to 10(4)copiesmL(-1) of the 16S rRNA genes, respectively. This resulted in the complete mineralization of 23μM 2,4,6-TBP within 35days (0.58μmolL(-1)d(-1)). Thus, using a synthetic microbial community of isolates or highly enriched cultures would be an efficient, optimizable, low-cost strategy for anaerobic bioremediation of halogenated aromatics.

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

    Directory of Open Access Journals (Sweden)

    M.T. Kato

    1997-12-01

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

  3. Anaerobic High-Throughput Cultivation Method for Isolation of Thermophiles Using Biomass-Derived Substrates

    Energy Technology Data Exchange (ETDEWEB)

    Hamilton-Brehm, Scott [ORNL; Vishnivetskaya, Tatiana A [ORNL; Allman, Steve L [ORNL; Mielenz, Jonathan R [ORNL; Elkins, James G [ORNL

    2012-01-01

    Flow cytometry (FCM) techniques have been developed for sorting mesophilic organisms, but the difficulty increases if the target microbes are thermophilic anaerobes. We demonstrate a reliable, high-throughput method of screening thermophilic anaerobic organisms using FCM and 96-well plates for growth on biomass-relevant substrates. The method was tested using the cellulolytic thermophiles Clostridium ther- mocellum (Topt = 55 C), Caldicellulosiruptor obsidiansis (Topt = 78 C) and the fermentative hyperthermo- philes, Pyrococcus furiosus (Topt = 100 C) and Thermotoga maritima (Topt = 80 C). Multi-well plates were incubated at various temperatures for approximately 72 120 h and then tested for growth. Positive growth resulting from single cells sorted into individual wells containing an anaerobic medium was verified by OD600. Depending on the growth substrate, up to 80 % of the wells contained viable cultures, which could be transferred to fresh media. This method was used to isolate thermophilic microbes from Rabbit Creek, Yellowstone National Park (YNP), Wyoming. Substrates for enrichment cultures including crystalline cellulose (Avicel), xylan (from Birchwood), pretreated switchgrass and Populus were used to cultivate organisms that may be of interest to lignocellulosic biofuel production.

  4. Feeding glycerol-enriched yeast culture improves lactation performance, energy status, and hepatic gluconeogenic enzyme expression of dairy cows during the transition period.

    Science.gov (United States)

    Ye, G; Liu, J; Liu, Y; Chen, X; Liao, S F; Huang, D; Huang, K

    2016-06-01

    This study aimed to evaluate the effects of feeding glycerol-enriched yeast culture (GY) on feed intake, lactation performance, blood metabolites, and expression of some key hepatic gluconeogenic enzymes in dairy cows during the transition period. Forty-four multiparous transition Holstein cows were blocked by parity, previous 305-d mature equivalent milk yield, and expected calving date and randomly allocated to 4 dietary treatments: Control (no additive), 2 L/d of GY (75.8 g/L glycerol and 15.3 g/L yeast), 150 g/d of glycerol (G; 0.998 g/g glycerol), and 1 L/d of yeast culture (Y; 31.1 g/L yeast). All additives were top-dressed and hand mixed into the upper one-third of the total mixed ration in the morning from -14 to +28 d relative to calving. Results indicated that the DMI, NE intake, change of BCS, and milk yields were not affected by the treatments ( > 0.05). Supplementation of GY or Y increased milk fat percentages, milk protein percentages, and milk protein yields relative to the Control or G group ( 0.10). In conclusion, dietary GY or Y supplementation increased the milk fat and protein content of the cows in early lactation and GY or G supplementation improved the energy status as indicated by greater plasma glucose and lower plasma BHBA and NEFA concentrations and upregulated the hepatic gluconeogenic enzymes of dairy cows during the transition period. Feeding cows with a GY mixture in the peripartum period combined the effects of yeast on lactation performance and the effects of glycerol on energy status in dairy cows.

  5. CD133-enriched Xeno-Free human embryonic-derived neural stem cells expand rapidly in culture and do not form teratomas in immunodeficient mice

    Directory of Open Access Journals (Sweden)

    Daniel L. Haus

    2014-09-01

    Full Text Available Common methods for the generation of human embryonic-derived neural stem cells (hNSCs result in cells with potentially compromised safety profiles due to maintenance of cells in conditions containing non-human proteins (e.g. in bovine serum or on mouse fibroblast feeders. Additionally, sufficient expansion of resulting hNSCs for scaling out or up in a clinically relevant time frame has proven to be difficult. Here, we report a strategy that produces hNSCs in completely “Xeno-Free” culture conditions. Furthermore, we have enriched the hNSCs for the cell surface marker CD133 via magnetic sorting, which has led to an increase in the expansion rate and neuronal fate specification of the hNSCs in vitro. Critically, we have also confirmed neural lineage specificity upon sorted hNSC transplantation into the immunodeficient NOD-scid mouse brain. The future use or adaptation of these protocols has the potential to better facilitate the advancement of pre-clinical strategies from the bench to the bedside.

  6. Contribution of enrichments and resampling for sulfate reducing bacteria diversity assessment by high-throughput cultivation.

    Science.gov (United States)

    Colin, Yannick; Goñi-Urriza, Marisol; Caumette, Pierre; Guyoneaud, Rémy

    2015-03-01

    The development of new high-throughput cultivation methods aims to increase the isolation efficiency as compared to standard techniques that often require enrichment procedures to compensate the low microbial recovery. In the current study, estuarine sulfate-reducing bacteria were isolated using an anaerobic isolation procedure in 384-well microplates. Ninety-nine strains were recovered from initial sediments. Isolates were identified according to their partial 16S rRNA sequences and clustered into 13 phylotypes. Besides, the increase in species richness obtained through enrichments or resampling was investigated. Forty-four enrichment procedures were conducted and shifts in sulfate-reducing bacterial communities were investigated through dsrAB gene fingerprinting. Despite efforts in conducting numerous enrichment conditions only few of them were statistically different from initial sample. The cultural diversity obtained from 3 of the most divergent enrichments, as well as from resampled sediments equally contributed to raise the sulfate-reducing diversity up to 22 phylotypes. Enrichments (selection of metabolism) or resampling (transient populations and micro-heterogeneity) may still be helpful to assess new microbial phylotypes. Nevertheless, all the newly cultivated strains were all representatives of minor Operational Taxonomic Units and could eventually be recovered by maintaining high-throughput isolation effort from the initial sediments.

  7. Anaerobic Digestion: Process

    DEFF Research Database (Denmark)

    Angelidaki, Irini; Batstone, Damien J.

    2011-01-01

    with very little dry matter may also be called a digest. The digest should not be termed compost unless it specifically has been composted in an aerated step. This chapter describes the basic processes of anaerobic digestion. Chapter 9.5 describes the anaerobic treatment technologies, and Chapter 9.......6 addresses the mass balances and environmental aspects of anaerobic digestion....

  8. Biohydrogen production by anaerobic fermentation of waste. Final project report

    Energy Technology Data Exchange (ETDEWEB)

    Karakashev, D.; Angelidaki, I.

    2009-01-15

    The objective of this project was to investigate and increase dark fermentative hydrogen production from organic wastes by optimizing important process parameters (reactor type, pH, temperature, organic loading, retention time, inoculation strategy, microbial composition). Labscale experiments were carried out at the Department of Environmental Engineering, Technical University of Denmark. A two steps process for hydrogen production in the first step and methane production in the second step in serial connected fully mixed reactors was developed and could successfully convert organic matter to approx. 20-25 % hydrogen and 15-80 % to methane. Sparging with methane produced in the second stage could significantly increase the hydrogen production. Additionally it was shown that upflow anaerobic sludge blanket (UASB) reactor system was very promising for high effective biohydrogen production from glucose at 70 deg C. Glucose-fed biofilm reactors filled with plastic carriers demonstrated high efficient extreme thermophilic biohydrogen production with mixed cultures. Repeated batch cultivations via exposure of the cultures to increased concentrations of household solid waste was found to be most useful method to enhance hydrogen production rate and reduce lag phase of extreme thermophilic fermentation process. Low level of pH (5.5) at 3-day HRT was enough to inhibit completely the methanogenesis and resulted in stable extreme thermophilic hydrogen production. Homoacetogenisis was proven to be an alternative competitor to biohydrogen production from organic acids under thermophilic (55 deg. C) conditions. With respect to microbiology, 16S rRNA targeted oligonucleotide probes were designed to monitor the spatial distribution of hydrogen producing bacteria in sludge and granules from anaerobic reactors. An extreme thermophilic (70 deg. C), strict anaerobic, mixed microbial culture with high hydrogen producing potential was enriched from digested household waste. Culture

  9. Effect of fish oil enriched enteral diet on inflammatory bowel disease tissues in organ culture: Differential effects on ulcerative colitis and Crohn's disease

    Institute of Scientific and Technical Information of China (English)

    Doris Meister; Subrata Ghosh

    2005-01-01

    AIM: To investigate the influence of fish oil enriched enteral diet on intestinal tissues taken from Crohn's disease (CD), ulcerative colitis (UC) and non-inflamed non-IBD control patients in vitro.METHODS: Colonoscopic biopsies from patients with active CD (n = 4), active UC (n = 7), and non-inflamed non-IBD control patients (n = 4) were incubated (three dilutions of 1:20, 1:10, and 1:5) with Waymouth's culture medium and enteral elemental diet (EO28, SHS, Liverpool, UK) modified in the fatty acid composition with fish oil (EF) in an organ culture system for 24 h. In each experimental set-up, incubation with Waymouth's medium alone as control was included. Tissue viability was assessed by adding bromodeoxyuridine (BrdU) to the culture fluid and immunohistochemically staining for BrdU uptake. Cytokine ratio of IL-1ra/IL-1β (low ratio indicative of inflammation) and production of those cytokines as a percentage of medium control were assayed in the culture supernatant. RESULTS: Incubation of CD-affected tissue with EF (1:20, 1:10, and 1:5) modestly and non-significantly increased IL-1ra/IL-1β ratio as compared with medium control (CD 39.1±16.1; 26.5±7.8, 47.1±16.8 vs control 13.0±2.2), but incubation of UC-affected tissues increased IL-1ra/IL-1β ratio significantly in all three dilutions (UC 69.1±32.2, P<0.05; 76.1±36.4, P = 0.05;84.5±37.3, P<0.02; vs control 10.2±3.7). Incubation of non-inflamed non-IBD control tissue did not increase the IL-1ra/IL-1β ratio in any dilution compared to medium control (69.3±47.0, 54.1±30.6, 79.4±34.0 vs control 76.1±37.3). Average percentage production of IL-1β indexed against medium control was significantly less in UC after EF incubation as compared with CD (UC 24.0±4.8 vs CD 51.8±8.1; P<0.05). Average percentage production of IL-1ra was markedly higher in UC (135.9±3.4) than that in control patients (36.5±4.3) (P<0.0001).CONCLUSION: IBD tissues, after incubation with elemental diet modified in its

  10. A comparative study on the effects of tylosin on select bacteria during continuous flow culture of mixed populations of gut microflora derived from a feral and a domestic pig.

    Science.gov (United States)

    Ramlachan, Nicole; Anderson, Robin C; Andrews, Kathleen; Harvey, Roger B; Nisbet, David J

    2008-02-01

    Continuous flow cultures of feral (culture FC) and domesticated (culture RPCF) pig gut microflora were established in steady state. Cultures were continuously infused with 25 or 100 microg tylosin/mL and sampled at intervals to assess effects on total culturable anaerobes, Bacteroides and Enterococcus via plating to agar supplemented without or with 100 microg tylosin/mL, the latter to assess bacterial sensitivity to tylosin. Endogenous tylosin-insensitive anaerobes within the cultures, while similar prior to tylosin administration, responded differently during tylosin administration, with concentrations in RPCF cultures becoming enriched more than in FC cultures. Tylosin-insensitive anaerobes in RPCF cultures persisted at increased concentrations after cessation of tylosin administration whereas concentrations in FC cultures decreased slightly. Concentrations of Bacteroides and endogenous Enterococcus recovered on medium without tylosin decreased to near or below detectable levels in FC cultures administered 25 or 100 microg tylosin/mL. Tylosin-insensitive Bacteroides were enriched to >5 log10 CFU/mL in RPCF cultures after 25 microg tylosin/mL but not at 100 microg tylosin/mL. Populations of endogenous tylosin-insensitive Enterococcus were enriched in RPCF but not FC cultures administered 25 or 100 microg tylosin/mL. In cultures administered 100 microg tylosin/mL, an exogenous-sourced E. faecium possessing tylosin resistance maintained itself only in the presence of tylosin. These results indicate that under the conditions of these tests, antibiotic exposure may enrich for antibiotic-insensitive bacteria populations of endogenous or exogenous origin but that the ability of an exogenous tylosin-resistant E. faecium to persist is reduced in the absence of the antibiotic, likely due to exclusion by native flora.

  11. Anaerobic Metabolism and Bioremediation of Explosives-Contaminated Soil

    Science.gov (United States)

    Boopathy, Raj

    Nitroaromatic compounds pollute soil, water, and food via use of pesticides, plastics, pharmaceuticals, landfill dumping of industrial wastes, and the military use of explosives. Biotransformation of trinitrotoluene and other nitroaromatics by aerobic bacteria in the laboratory has been frequently reported, but the anaerobic bacterial metabolism of nitroaromatics has not been studied as extensively perhaps due to the difficulty in working with anaerobic cultures and the slow growth of anaerobes. Sulfate-reducing and methanogenic bacteria can metabolize nitroaromatic compounds under anaerobic conditions if appropriate electron donors and electron acceptors are present in the environment.

  12. A combined approach for the enhanced detection and isolation of Bartonella species in dog blood samples: pre-enrichment liquid culture followed by PCR and subculture onto agar plates.

    Science.gov (United States)

    Duncan, Ashlee W; Maggi, Ricardo G; Breitschwerdt, Edward B

    2007-05-01

    Historically, direct plating, lysis centrifugation, or freeze-thaw approaches have proven to be highly insensitive methods for confirming Bartonella species infection in dogs. A prospective study was designed to compare diagnostic methods for the detection of Bartonella using samples submitted to the Vector-Borne Disease Diagnostic Laboratory at North Carolina State University. Methods included indirect immunofluorescence assay, PCR, direct inoculation of a blood agar plate (trypticase soy agar with 5% rabbit blood), and inoculation into a novel pre-enrichment liquid medium, Bartonella/alpha-Proteobacteria growth medium (BAPGM). Sequential research efforts resulted in the development of a combinational approach consisting of pre-enrichment culture of Bartonella species in BAPGM, sub-inoculation of the liquid culture onto agar plates, followed by DNA amplification using PCR. The multi-faceted approach resulted in substantial improvement in the microbiological detection and isolation of Bartonella when compared to direct inoculation of a blood agar plate. Importantly, this approach facilitated the detection and subsequent isolation of both single and co-infections with two Bartonella species in the blood of naturally infected dogs. The use of a combinational approach of pre-enrichment culture and PCR may assist in the diagnostic confirmation of bartonellosis in dogs and other animals.

  13. The effects of co-contaminants and native wetland sediments on the activity and dominant transformation mechanisms of a 1,1,2,2-tetrachloroethane (TeCA)-degrading enrichment culture

    Science.gov (United States)

    Lorah, Michelle M.; Schiffmacher, Emily N.; Becker, Jennifer G.; Voytek, Mary A.

    2016-01-01

    Bioremediation strategies, including bioaugmentation with chlorinated ethene-degrading enrichment cultures, have been successfully applied in the cleanup of subsurface environments contaminated with tetrachloroethene (PCE) and/or trichloroethene (TCE). However, these compounds are frequently found in the environment as components of mixtures that may also contain chlorinated ethanes and methanes. Under these conditions, the implementation of bioremediation may be complicated by inhibition effects, particularly when multiple dehalorespirers are present. We investigated the ability of the 1,1,2,2-tetrachloroethane (TeCA)-dechlorinating culture WBC-2 to biotransform TeCA alone, or a mixture of TeCA plus PCE and carbon tetrachloride (CT), in microcosms. The microcosms contained electron donors provided to biostimulate the added culture and sediment collected from a wetland where numerous “hotspots” of contamination with chlorinated solvent mixtures exist. The dominant TeCA biodegradation mechanism mediated by the WBC-2 culture in the microcosms was different in the presence of these wetland sediments than in the sediment-free enrichment culture or in previous WBC-2 bioaugmented microcosms and column tests conducted with wetland sediment collected at nearby sites. The co-contaminants and their daughter products also inhibited TeCA biodegradation by WBC-2. These results highlight the need to conduct biodegradability assays at new sites, particularly when multiple contaminants and dehalorespiring populations are present.

  14. Anaerobic filters for the treatment of coal gasification wastewater.

    Science.gov (United States)

    Suidan, M T; Siekerka, G L; Kao, S W; Pfeffer, J T

    1983-06-01

    A process train consisting of the following sequence of unit processes, a berl-saddle-packed anaerobic filter, an expanded bed, granular activated carbon anaerobic filter, and an activated sludge nitrification system was evaluated for the treatment of a synthetically prepared coal gasification wastewater. The first-stage anaerobic filter resulted in very little removal of organic matter and no methane production. Excellent reduction in organic matter occurred in the granular activated carbon anaerobic filter. The removal mechanism was initially adsorptive and near the end of the study, removal of organic matter was primarily through conversion to methane gas. It is felt that the success of the activated carbon anaerobic filter was due to the ability of the activated carbon to sequester some components of the wastewater that were toxic to the mixed culture of anaerobic microorganisms. The activated sludge nitrification system resulted in complete ammonia oxidation and was very efficient in final effluent polishing.

  15. Rapid identification of Salmonella serovars in feces by specific detection of virulence genes, invA and spvC, by an enrichment broth culture-multiplex PCR combination assay.

    Science.gov (United States)

    Chiu, C H; Ou, J T

    1996-10-01

    In order to make a rapid and definite diagnosis of Salmonella enteritis in children, an enrichment broth culture-multiplex PCR combination assay was devised to identify Salmonella serovars directly from fecal samples. Two pairs of oligonucleotide primers were prepared according to the sequences of the chromosomal invA and plasmid spvC genes. PCR with these two primers would produce either one amplicon (from the invA gene) or two amplicons (from the invA and spvC genes), depending on whether or not the Salmonella bacteria contained a virulence plasmid. The fecal sample was diluted 10- to 20-fold into gram-negative enrichment broth and incubated to eliminate inhibitory compounds and also to allow selective enrichment of the bacteria. One or two amplicons were obtained, the expected result if Salmonella bacteria were present. The detection limit of this PCR was about 200 bacteria per reaction mixture. The primers were specific, as no amplification products were obtained with 18 species and 22 isolates of non-Salmonella bacteria tested which could be present in the feces or cause contamination. In contrast, when 23 commonly seen Salmonella serovars (38 isolates) were tested, all were shown to carry the invA gene and seven concomitantly harbored the spvC gene of the virulence plasmid. This assay was applied to the diagnosis of Salmonella enteritis in 57 children who were suffering from mucoid and/or bloody diarrhea. Of the 57 children, 38 were PCR positive and 22 were culture positive. There were two culture-positive samples that were not detected by PCR. Thus, this PCR assay showed an efficiency of 95% (38 of 40), which is much higher than the 60% (24 of 40) by culture alone. Not only is this method more sensitive, rapid, and efficient but it will cause only an incremental increase in the cost of stool processing, since enrichment cultivation of fecal samples from diarrheal patients using gram-negative enrichment broth is a routine practice for identification in many

  16. Direct identification of bacteria from BacT/ALERT anaerobic positive blood cultures by MALDI-TOF MS: MALDI Sepsityper kit versus an in-house saponin method for bacterial extraction.

    Science.gov (United States)

    Meex, Cécile; Neuville, Florence; Descy, Julie; Huynen, Pascale; Hayette, Marie-Pierre; De Mol, Patrick; Melin, Pierrette

    2012-11-01

    In cases of bacteraemia, a rapid species identification of the causal agent directly from positive blood culture broths could assist clinicians in the timely targeting of empirical antimicrobial therapy. For this purpose, we evaluated the direct identification of micro-organisms from BacT/ALERT (bioMérieux) anaerobic positive blood cultures without charcoal using the Microflex matrix-assisted laser desorption/ionization (MALDI) time of flight MS (Bruker), after bacterial extraction by using two different methods: the MALDI Sepsityper kit (Bruker) and an in-house saponin lysis method. Bruker's recommended criteria for identification were expanded in this study, with acceptance of the species identification when the first three results with the best matches with the MALDI Biotyper database were identical, whatever the scores were. In total, 107 monobacterial cultures and six polymicrobial cultures from 77 different patients were included in this study. Among monomicrobial cultures, we identified up to the species level 67 and 66 % of bacteria with the MALDI Sepsityper kit and the saponin method, respectively. There was no significant difference between the two extraction methods. The direct species identification was particularly inconclusive for Gram-positive bacteria, as only 58 and 52 % of them were identified to the species level with the MALDI Sepsityper kit and the saponin method, respectively. Results for Gram-negative bacilli were better, with 82.5 and 90 % of correct identification to the species level with the MALDI Sepsityper kit and the saponin method, respectively. No misidentifications were given by the direct procedures when compared with identifications provided by the conventional method. Concerning the six polymicrobial blood cultures, whatever the extraction method used, a correct direct identification was only provided for one of the isolated bacteria on solid medium in all cases. The analysis of the time-to-result demonstrated a reduction

  17. 东南太平洋热液区硫化物样品富集培养物中Fe3+还原细菌的多样性分析%Diversity analysis of Fe3+ -reducing bacteria in enriched cultures from hydrothermal sulfide sample of southeast Pacific Ocean

    Institute of Scientific and Technical Information of China (English)

    秦丹; 张晓波; 姜丽晶; 盖英宝; 曾湘

    2011-01-01

    采集了东南太平洋热液区DY115-20V-S35-TVG17站位的沉积物硫化物样品,其含有76.717% (m/m)强还原性的磁性良好的黄铁矿.以乳酸钠为电子供体,不溶性的氢氧化铁为电子受体,对该样品进行Fe3+还原细菌的定向富集培养.通过构建该菌群的16S rDNA基因文库,随机挑选82个阳性克隆子进行群落结构多样性分析.研究结果表明:82个克隆代表的6种基因型分别属于4个主要类群,其中ε-变形杆菌占有绝对优势,达到55.2%.此外其还包括厚壁菌门、β-变形杆菌和α-变形杆菌,克隆子比例依次为27.6%、10.3%、6.9%.在富集产物中还原氢氧化铁的功能菌群主要是具有硫还原作用的ε-变形菌中的硫还原菌.%The ocean contains rich resources and in deep sea sediments there exists iron compounds at different stages of oxidation and reduction. Iron is one of the most widely distributed elements on the earth and its cycle chiefly includes two processes of oxidation and reduction. As a primitive form of respiration, Fe3 + -reduction and Fe3 + -reducing bacteria play an important role in the iron cycle where in sediments with an anaerobic environment, where iron chiefly exists in a reduced form. In the current study, a hydrothermal sulfide sample was obtained from the southeast Pacific Ocean, site DY115-20V-S35-TVG17 (50.648 l°N,37.780 2°W, depth 2 783 m) using a television grasp sampler in December, 2008, and was found to be mainly composed of iron pyrite with good magnetism, chiefly Fe1-XS and FeS2, 76.717% (m/m) through X-ray detection. To investigate the diversity of Fe3 +-reducing bacteria in the sample, we designed a medium with sodium lactate as the electron donor and insoluble ferric hydroxide as electron acceptor and cultivated the enriched culture of Fe3 +-reducing bacteria from this sample. The 16S Rdna gene library showed that the 29 selected clones in enriched cultures were clustered into two groups. Pro

  18. Use of starter cultures of lactic acid bacteria and yeasts as inoculum enrichment for the production of gowé, a sour beverage from Benin

    DEFF Research Database (Denmark)

    Vieira-Dalodé, G.; Madodé, Y.E.; Hounhouigan, J.

    2008-01-01

    Lactobacillus fermentum, Weissella confusa, Kluyveromyces marxianus and Pichia anomala, previously isolated during natural fermentation of traditional gowé, were tested as inoculum enrichment for controlled fermentation of gowé. The final product was subjected to chemical analysis and sensory eva...

  19. Phosphorus uptake by potato from fertilizers recovered from anaerobic digestion

    Science.gov (United States)

    Field experiments were conducted in the Columbia Basin of South Central Washington to assess the yield of potato (Solanum tuberosum) in response to application of phosphorus enriched materials recovered from anaerobic digestion of manure. The treatments were comprised of four rates (0, 56, 112 and ...

  20. In vitro Characterization of Phenylacetate Decarboxylase, a Novel Enzyme Catalyzing Toluene Biosynthesis in an Anaerobic Microbial Community

    DEFF Research Database (Denmark)

    Zargar, K.; Saville, R.; Phelan, R. M.;

    2016-01-01

    an anaerobic, sewage-derived enrichment culture that quantitatively produces toluene from phenylacetate; complementary metagenomic and metaproteomic analyses are also presented. Among the noteworthy findings is that this enzyme is not the well-characterized clostridial p-hydroxyphenylacetate decarboxylase (Csd...... similarly inhibited by an amide analog of p-hydroxyphenylacetate. Based upon these and other data, we hypothesize that the toluene synthase reaction involves a glycyl radical decarboxylase. This first-time study of the phenylacetate decarboxylase reaction constitutes an important step in understanding...

  1. Anaerobic Digestion and its Applications

    Science.gov (United States)

    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...

  2. Multiplex Real-Time PCR Assays for Screening of Shiga Toxin 1 and 2 Genes, Including All Known Subtypes, and Escherichia coli O26-, O111-, and O157-Specific Genes in Beef and Sprout Enrichment Cultures.

    Science.gov (United States)

    Harada, Tetsuya; Iguchi, Atsushi; Iyoda, Sunao; Seto, Kazuko; Taguchi, Masumi; Kumeda, Yuko

    2015-10-01

    Shiga toxin family members have recently been classified using a new nomenclature into three Stx1 subtypes (Stx1a, Stx1c, and Stx1d) and seven Stx2 subtypes (Stx2a, Stx2b, Stx2c, Stx2d, Stx2e, Stx2f, and Stx2g). To develop screening methods for Stx genes, including all of these subtype genes, and Escherichia coli O26-, O111-, and O157-specific genes in laboratory investigations of Shiga toxin-producing E. coli (STEC) foodborne cases, we developed multiplex real-time PCR assays and evaluated their specificity and quantitative accuracy using STEC and non-STEC isolates, recombinant plasmids, and food enrichment cultures and by performing STEC spiking experiments with beef and sprout enrichment cultures. In addition, we evaluated the relationship between the recovery rates of the target strains by direct plating and immunomagnetic separation and the cycle threshold (CT) values of the real-time PCR assays for the Stx subtypes and STEC O26, O111, and O157 serogroups. All three stx1- and seven stx2-subtype genes were detected by real-time PCR with high sensitivity and specificity, and the quantitative accuracy of this assay was confirmed using control plasmids and STEC spiking experiments. The results of the STEC spiking experiments suggest that it is not routinely possible to isolate STEC from enrichment cultures with real-time PCR CT values greater than 30 by direct plating on MacConkey agar, although highly selective media and immunomagnetic beads were able to isolate the inoculated strains from the enrichment cultures. These data suggest that CT values obtained from the highly quantitative real-time PCR assays developed in this study provide useful information to develop effective isolation strategies for STEC from food samples. The real-time PCR assays developed here are expected to aid in investigations of infections or outbreaks caused by STEC harboring any of the stx-subtype genes in the new Stx nomenclature, as well as STEC O26, O111, and O157.

  3. Ubiquitous Presence and Novel Diversity of Anaerobic Alkane Degraders in Cold Marine Sediments.

    Science.gov (United States)

    Gittel, Antje; Donhauser, Johanna; Røy, Hans; Girguis, Peter R; Jørgensen, Bo B; Kjeldsen, Kasper U

    2015-01-01

    Alkanes are major constituents of crude oil and are released to the marine environment by natural seepage and from anthropogenic sources. Due to their chemical inertness, their removal from anoxic marine sediments is primarily controlled by the activity of anaerobic alkane-degrading microorganisms. To facilitate comprehensive cultivation-independent surveys of the diversity and distribution of anaerobic alkane degraders, we designed novel PCR primers that cover all known diversity of the 1-methylalkyl succinate synthase gene (masD/assA), which catalyzes the initial activation of alkanes. We studied masD/assA gene diversity in pristine and seepage-impacted Danish coastal sediments, as well as in sediments and alkane-degrading enrichment cultures from the Middle Valley (MV) hydrothermal vent system in the Pacific Northwest. MasD/assA genes were ubiquitously present, and the primers captured the diversity of both known and previously undiscovered masD/assA gene diversity. Seepage sediments were dominated by a single masD/assA gene cluster, which is presumably indicative of a substrate-adapted community, while pristine sediments harbored a diverse range of masD/assA phylotypes including those present in seepage sediments. This rare biosphere of anaerobic alkane degraders will likely increase in abundance in the event of seepage or accidental oil spillage. Nanomolar concentrations of short-chain alkanes (SCA) were detected in pristine and seepage sediments. Interestingly, anaerobic alkane degraders closely related to strain BuS5, the only SCA degrader in pure culture, were found in mesophilic MV enrichments, but not in cold sediments from Danish waters. We propose that the new masD/assA gene lineages in these sediments represent novel phylotypes that are either fueled by naturally occurring low levels of SCA or that metabolize medium- to long-chain alkanes. Our study highlights that masD/assA genes are a relevant diagnostic marker to identify seepage and microseepage, e

  4. Antipathogenic activity of probiotics against Salmonella Typhimurium and Clostridium difficile in anaerobic batch culture systems: is it due to synergies in probiotic mixtures or the specificity of single strains?

    Science.gov (United States)

    Tejero-Sariñena, Sandra; Barlow, Janine; Costabile, Adele; Gibson, Glenn R; Rowland, Ian

    2013-12-01

    Probiotics are currently being investigated for prevention of infections caused by enteric pathogens. The aim of this in vitro study was to evaluate the influence of three single probiotics: Lactobacillus casei NCIMB 30185 (PXN 37), Lactobacillus acidophilus NCIMB 30184 (PXN 35), Bifidobacterium breve NCIMB 30180 (PXN 25) and a probiotic mixture containing the above strains plus twelve other strains belonging to the Lactobacillus, Bifidobacterium, Lactococcus, Streptococcus and Bacillus genera on the survival of Salmonella Typhimurium and Clostridium difficile using pH-controlled anaerobic batch cultures containing mixed faecal bacteria. Changes in relevant bacterial groups and effects of probiotic addition on survival of the two pathogens were assessed over 24 h. Quantitative analysis of bacterial populations revealed that there was a significant increase in lactobacilli and/or bifidobacteria numbers, depending on probiotic addition, compared with the control (no added probiotic). There was also a significant reduction in S. Typhimurium and C. difficile numbers in the presence of certain probiotics compared with controls. Of the probiotic treatments, two single strains namely L. casei NCIMB 30185 (PXN 37), and B. breve NCIMB 30180 (PXN 25) were the most potent in reducing the numbers of S. Typhimurium and C. difficile. In addition, the supplementation with probiotics into the systems influenced some fermentations parameters. Acetate was found in the largest concentrations in all vessels and lactate and formate were generally detected in higher amounts in vessels with probiotic addition compared to controls.

  5. Anaerobic Digestion Foaming Causes

    OpenAIRE

    Ganidi, Nafsika

    2008-01-01

    Anaerobic digestion foaming has been encountered in several sewage treatment plants in the UK. Foaming has raised major concerns for the water utilities due to significant impacts on process efficiency and operational costs. Several foaming causes have been suggested over the past few years by researchers. However, the supporting experimental information is limited and in some cases site specific. The present report aimed to provide a better understanding of the anaerobic di...

  6. Anaerobic infections in the head and neck region.

    Science.gov (United States)

    Tabaqchali, S

    1988-01-01

    Anaerobic bacteria form the predominant flora of the oral cavity, outnumbering facultative organisms by 10-1,000: 1. The type of anaerobic bacteria and their concentration depend on the anatomical site and the degree of anaerobiosis in the different sites in the mouth. Three groups of anaerobic bacteria inhabit the oral cavity; the strict anaerobes, the moderate anaerobes, and the microaerophilic group of organisms. The majority of anaerobic bacterial infections occurring in the region of the mouth, head and neck are caused by the commensal flora. These infections include dental and periodontal disease where the predominant organisms are Bacteroides species, Veillonella, Bifidobacteria, Peptococcus, Peptostreptococcus and Propionibacterium species. More recently, Bacteroides endontalis has been isolated from a periapical abscess of endodontal origin and B. gingivalis, B. intermedius, Haemophilus actinomycetemcomitans and Wollinella species in chronic periodontal disease. Treponema species and other strict anaerobes are seen in smears of severe periodontal disease and acute necrotising gingivitis, but have not yet been isolated in pure culture. Until such time, their role in disease remains uncertain. Fusobacterium nucleatum is specially associated with severe orofacial infections which may extend into the mediastinum. Other anaerobic infections include chronic otitis media, chronic sinusitis and mastoiditis, and brain abscess. Treatment of these conditions should include the use of beta-lactamase resistant antimicrobials, such as clindamycin or one of the nitroimidazoles with penicillin.

  7. Methane and hydrogen production by human intestinal anaerobic bacteria.

    Science.gov (United States)

    McKay, L F; Holbrook, W P; Eastwood, M A

    1982-06-01

    The gas above liquid cultures of a variety of human intestinal anaerobic bacteria was sampled and analysed by headspace gas chromatography. Hydrogen production was greatest with strains of the genus Clostridium, intermediate with anaerobic cocci and least with Bacteroides sp. Very few strains produced methane although small amounts were detected with one strain of B. thetaiotaomicron, C. perfringens and C. histolyticum. There may be a relationship between these anaerobic bacteria and several gastrointestinal disorders in which there is a build up of hydrogen or methane in the intestines.

  8. Anaerobic Biodegradation of Detergent Surfactants

    Directory of Open Access Journals (Sweden)

    Erich Jelen

    2009-03-01

    Full Text Available Detergent surfactantscan 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. Microbiological Hydrogen Production by Anaerobic Fermentation and Photosynthetic Process

    Energy Technology Data Exchange (ETDEWEB)

    Asada, Y.; Ohsawa, M.; Nagai, Y.; Fukatsu, M.; Ishimi, K.; Ichi-ishi, S.

    2009-07-01

    Hydrogen gas is a clean and renewable energy carrier. Microbiological hydrogen production from glucose or starch by combination used of an anaerobic fermenter and a photosynthetic bacterium, Rhodobacter spheroides RV was studied. In 1984, the co-culture of Clostridium butyricum and RV strain to convert glucose to hydrogen was demonstrated by Miyake et al. Recently, we studied anaerobic fermentation of starch by a thermophilic archaea. (Author)

  10. Taurine reduction in anaerobic respiration of Bilophila wadsworthia RZATAU.

    Science.gov (United States)

    Laue, H; Denger, K; Cook, A M

    1997-05-01

    Organosulfonates are important natural and man-made compounds, but until recently (T. J. Lie, T. Pitta, E. R. Leadbetter, W. Godchaux III, and J. R. Leadbetter. Arch. Microbiol. 166:204-210, 1996), they were not believed to be dissimilated under anoxic conditions. We also chose to test whether alkane- and arenesulfonates could serve as electron sinks in respiratory metabolism. We generated 60 anoxic enrichment cultures in mineral salts medium which included several potential electron donors and a single organic sulfonate as an electron sink, and we used material from anaerobic digestors in communal sewage works as inocula. None of the four aromatic sulfonates, the three unsubstituted alkanesulfonates, or the N-sulfonate tested gave positive enrichment cultures requiring both the electron donor and electron sink for growth. Nine cultures utilizing the natural products taurine, cysteate, or isethionate were considered positive for growth, and all formed sulfide. Two clearly different pure cultures were examined. Putative Desulfovibrio sp. strain RZACYSA, with lactate as the electron donor, utilized sulfate, aminomethanesulfonate, taurine, isethionate, and cysteate, converting the latter to ammonia, acetate, and sulfide. Strain RZATAU was identified by 16S rDNA analysis as Bilophila wadsworthia. In the presence of, e.g., formate as the electron donor, it utilized, e.g., cysteate and isethionate and converted taurine quantitatively to cell material and products identified as ammonia, acetate, and sulfide. Sulfite and thiosulfate, but not sulfate, were utilized as electron sinks, as was nitrate, when lactate was provided as the electron donor and carbon source. A growth requirement for 1,4-naphthoquinone indicates a menaquinone electron carrier, and the presence of cytochrome c supports the presence of an electron transport chain. Pyruvate-dependent disappearance of taurine from cell extracts, as well as formation of alanine and release of ammonia and acetate, was

  11. Invited review: anaerobic fermentation of dairy food wastewater.

    Science.gov (United States)

    Hassan, A N; Nelson, B K

    2012-11-01

    Dairy food wastewater disposal represents a major environmental problem. This review discusses microorganisms associated with anaerobic digestion of dairy food wastewater, biochemistry of the process, factors affecting anaerobic digestion, and efforts to develop defined cultures. Anaerobic digestion of dairy food wastewater offers many advantages over other treatments in that a high level of waste stabilization is achieved with much lower levels of sludge. In addition, the process produces readily usable methane with low nutrient requirements and no oxygen. Anaerobic digestion is a series of complex reactions that broadly involve 2 groups of anaerobic or facultative anaerobic microorganisms: acidogens and methanogens. The first group of microorganisms breaks down organic compounds into CO(2) and volatile fatty acids. Some of these organisms are acetogenic, which convert long-chain fatty acids to acetate, CO(2), and hydrogen. Methanogens convert the acidogens' products to methane. The imbalance among the different microbial groups can lead not only to less methane production, but also to process failure. This is due to accumulation of intermediate compounds, such as volatile fatty acids, that inhibit methanogens. The criteria used for evaluation of the anaerobic digestion include levels of hydrogen and volatile fatty acids, methane:carbon ratio, and the gas production rate. A steady state is achieved in an anaerobic digester when the pH, chemical oxygen demand of the effluent, the suspended solids of the effluent, and the daily gas production remain constant. Factors affecting efficiency and stability of the process are types of microorganisms, feed C:N ratio, hydraulic retention time, reactor design, temperature, pH control, hydrogen pressure, and additives such as manure and surfactants. As anaerobic digesters become increasingly used in dairy plants, more research should be directed toward selecting the best cultures that maximize methane production from dairy

  12. ISOLATION OF ANAEROBES IN DEEP SEATED PRESSURE ULCERS USING A NOVEL INNOVATIVE TECHNIQUE OF ANAEROBE ISOLATION

    Directory of Open Access Journals (Sweden)

    Lalbiaktluangi

    2015-12-01

    Full Text Available BACKGROUND Isolation of an anaerobe is usually neglected in hospitals with limited resources due to the expensive and complicated technique of anaerobic isolation methods, which is difficult to arrange in such resource poor settings. Conventionally adopted anaerobic culture methods such as Anaerobic jar, Gas-Pak, Anoxomat or Automated glove-box systems are extremely costly and cumbersome for single unit testing, but not suitable for small scale laboratories. However, anaerobic bacteria are not to be overlooked as they have made a comeback in clinical settings and are even showing resistance to Metronidazole, once thought to be the gold standard bullet against anaerobes. Deep seated pressure ulcers are usually the site where anaerobe causes an infection in synergy with aerobes. AIMS AND OBJECTIVES Isolation of anaerobes in deep seated pressure ulcers using a novel innovative technique and to study their antibiogram profile. MATERIALS AND METHODS Swabs taken from depth of deep seated pressure ulcers were immediately inoculated in Brucella blood agar at bedside and placed in polycarbonate airtight jar for anaerobic incubation using a novel innovative Modified Candle Jar technique. In this technique five grams of grease-free grade zero steel wool were dipped in 50ml freshly prepared acidified copper sulphate solution until the copper colour appeared. Excess solution was drained and the steel wool was moulded into a loose pad to fit on an open Petri plate placed on top of the inoculated Brucella blood agar plates. A white-wax candle was placed at the centre of this plate. A small test tube containing mixture of 0.5g sodium-bicarbonate and 0.5g magnesium carbonate was kept ready to be placed inside the jar, just after placing the inoculated plate and incubated for 48 hours. RESULTS Peptostreptococcus anaerobius and Bacteroides fragilis were successfully isolated from deep seated pressure ulcers by this method. Antibiogram studies were done using the

  13. Metabolic capabilities of microorganisms involved in and associated with the anaerobic oxidation of methane

    Directory of Open Access Journals (Sweden)

    Gunter eWegener

    2016-02-01

    Full Text Available In marine sediments the anaerobic oxidation of methane with sulfate as electron acceptor (AOM is responsible for the removal of a major part of the greenhouse gas methane. AOM is performed by consortia of anaerobic methane-oxidizing archaea (ANME and their specific partner bacteria. The physiology of these organisms is poorly understood, which is due to their slow growth with doubling times in the order of months and the phylogenetic diversity in natural and in vitro AOM enrichments. Here we study sediment-free long-term AOM enrichments that were cultivated from seep sediments sampled off the Italian Island Elba (20°C; hereon called E20 and from hot vents of the Guaymas Basin, Gulf of California, cultivated at 37°C (G37 or at 50°C (G50. These enrichments were dominated by consortia of ANME-2 archaea and Seep-SRB2 partner bacteria (E20 or by ANME-1, forming consortia with Seep-SRB2 bacteria (G37 or with bacteria of the HotSeep-1 cluster (G50. We investigate lipid membrane compositions as possible factors for the different temperature affinities of the different ANME clades and show autotrophy as characteristic feature for both ANME clades and their partner bacteria. Although in the absence of additional substrates methane formation was not observed, methanogenesis from methylated substrates (methanol and methylamine could be quickly stimulated in the E20 and the G37 enrichment. Responsible for methanogenesis are archaea from the genus Methanohalophilus and Methanococcoides, which are minor community members during AOM (1 to 7‰ of archaeal 16S rRNA gene amplicons. In the same two cultures also sulfur disproportionation could be quickly stimulated by addition of zero-valent colloidal sulfur. The isolated partner bacteria are likewise minor community members (1 to 9‰ of bacterial 16S rRNA gene amplicons, whereas the dominant partner bacteria (Seep-SRB1a, Seep-SRB2 or HotSeep-1 did not grow on elemental sulfur. Our results support a

  14. Effect of a Semi-Purified Oligosaccharide-Enriched Fraction from Caprine Milk on Barrier Integrity and Mucin Production of Co-Culture Models of the Small and Large Intestinal Epithelium

    Directory of Open Access Journals (Sweden)

    Alicia M. Barnett

    2016-05-01

    Full Text Available Caprine milk contains the highest amount of oligosaccharides among domestic animals, which are structurally similar to human milk oligosaccharides (HMOs. This suggests caprine milk oligosaccharides may offer similar protective and developmental effects to that of HMOs. However, to date, studies using oligosaccharides from caprine milk have been limited. Thus, this study aimed to examine the impact of a caprine milk oligosaccharide-enriched fraction (CMOF on barrier function of epithelial cell co-cultures of absorptive enterocytes (Caco-2 cells and mucus-secreting goblet cells (HT29-MTX cells, that more closely simulate the cell proportions found in the small (90:10 and large intestine (75:25. Treatment of epithelial co-cultures with 0.4, 1.0, 2.0 and 4.0 mg/mL of CMOF was shown to have no effect on metabolic activity but did enhance cell epithelial barrier integrity as measured by trans-epithelial electrical resistance (TEER, in a dose-dependent manner. The CMOF at the maximum concentration tested (4.0 mg/mL enhanced TEER, mucin gene expression and mucin protein abundance of epithelial co-cultures, all of which are essential components of intestinal barrier function.

  15. Anaerobic degradation of long-chain alkylamines by a denitrifying Pseudomonas stutzeri.

    Science.gov (United States)

    Nguyen, Phuong D; van Ginkel, Cornelis G; Plugge, Caroline M

    2008-10-01

    The anaerobic degradation of tetradecylamine and other long-chain alkylamines by a newly isolated denitrifying bacterium was studied. Strain ZN6 was isolated from a mixture of soil and active sludge and was identified as representing Pseudomonas stutzeri, based on partial 16S rRNA gene sequence analysis. Strain ZN6 was a mesophilic, motile, Gram-negative rod-shaped bacterium and was able to grow on a variety of compounds including even-numbered primary fatty amines with alkyl chains ranging from C(4) to C(18) coupled to nitrate reduction. Alkylamines were used as sole carbon, energy and nitrogen source and were completely mineralized. Nitrate was dissimilated by ZN6 to nitrite. When strain ZN6 was grown under nitrate limitation, nitrite was slowly dissimilated further. When cocultivated with the complete denitrifier Castellaniella defragens ZN3, anaerobic degradation under denitrifying of alkylamines by strain ZN6 was slightly faster. Strain ZN3 is a complete denitrifier, unable to convert tetradecylamine, and was copurified from the same enrichment culture as strain ZN6. The proposed pathway for the degradation of alkylamines in strain ZN6 starts with C-N cleavages to alkanals and further oxidation to the corresponding fatty acids.

  16. Importance of cobalt for individual trophic groups in an anaerobic methanol-degrading consortium.

    OpenAIRE

    Florencio, L; Field, J A; Lettinga, G

    1994-01-01

    Methanol is an important anaerobic substrate in industrial wastewater treatment and the natural environment. Previous studies indicate that cobalt greatly stimulates methane formation during anaerobic treatment of methanolic wastewaters. To evaluate the effect of cobalt in a mixed culture, a sludge with low background levels of cobalt was cultivated in an upflow anaerobic sludge blanket reactor. Specific inhibitors in batch assays were then utilized to study the effect of cobalt on the growth...

  17. Anaerobic treatability of wastewater contaminated with propylene glycol.

    Science.gov (United States)

    Sezgin, Naim; Tonuk, Gulseven Ubay

    2013-09-01

    The purpose of this study was to investigate the biodegradability of propylene glycol in anaerobic conditions by using methanogenic culture. A master reactor was set up to develop a culture that would be acclimated to propylene glycol. After reaching steady-state, culture was transferred to serum bottles. Three reactors with same initial conditions were run for consistency. Propylene glycol was completely biodegradable under anaerobic methanogenic conditions. Semi-continuous reactors operated at a temperature of 35°C had consistently achieved a propylene glycol removal of higher than 95 % based on chemical oxygen demand (COD). It was found that in semi-continuous reactors, anaerobic treatment of propylene glycol at concentrations higher than 1,500 mg COD m(-3) day(-1) was not convenient due to instable effluent COD.

  18. The anaerobic digestion process

    Energy Technology Data Exchange (ETDEWEB)

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

    1996-01-01

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

  19. Anaerobic digestion without biogas?

    NARCIS (Netherlands)

    Kleerebezem, R.; Joosse, B.; Rozendaal, R.; Van Loosdrecht, M.C.M.

    2015-01-01

    Anaerobic digestion for the production of methane containing biogas is the classic example of a resource recovery process that combines stabilization of particulate organic matter or wastewater treatment with the production of a valuable end-product. Attractive features of the process include the pr

  20. Prospective study of the clinical performance of three BACTEC media in a modern emergency department: Plus Aerobic/F, Plus Anaerobic/F, and Anaerobic Lytic/F.

    Science.gov (United States)

    Rocchetti, Andrea; Di Matteo, Luigi; Bottino, Paolo; Foret, Benjamin; Gamalero, Elisa; Calabresi, Alessandra; Guido, Gianluca; Casagranda, Ivo

    2016-11-01

    The performance of 3 blood culture bottles (BACTEC Plus Aerobic/F, Plus Anaerobic/F, and Anaerobic Lytic/F) were analyzed with clinical specimens collected from 688 Emergency Department patients. A total of 270 strains belonging to 33 species were identified, with E. coli and S. aureus as the most frequently detected. Overall recovery rate (RR) of bacteria and yeast was equivalent in the Plus Aerobic/F vials (208 of 270 isolates; 77.0%) and Anaerobic Lytic/F vials (206 isolates; 76.3%) and significantly better than in the Plus Anaerobic/F vials (189 isolates; 70.0%). Median time to detection (TTD) was earliest with the Anaerobic Lytic/F vials (12.0h) compared with the Plus Aerobic/F (14.6h) and Plus Anaerobic/F vials (15.4h). Positivity rate (PR) was similar for Anaerobic Lytic/F vials (76.9%) and Plus Aerobic/F vials (76.5%), but better if compared with Plus Anaerobic/F vials (69.4%). The PR and TTD for the combination of Plus Aerobic/F with Anaerobic Lytic/F (94.5% and 12.3h, respectively) was significantly better than with Plus Aerobic/F with Plus Anaerobic/F (87.8% and 14.1h).

  1. Anaerobic benzene oxidation via phenol in Geobacter metallireducens.

    Science.gov (United States)

    Zhang, Tian; Tremblay, Pier-Luc; Chaurasia, Akhilesh Kumar; Smith, Jessica A; Bain, Timothy S; Lovley, Derek R

    2013-12-01

    Anaerobic activation of benzene is expected to represent a novel biochemistry of environmental significance. Therefore, benzene metabolism was investigated in Geobacter metallireducens, the only genetically tractable organism known to anaerobically degrade benzene. Trace amounts (Geobacter metallireducens anaerobically oxidizing benzene to carbon dioxide with the reduction of Fe(III). Phenol was not detected in cell-free controls or in Fe(II)- and benzene-containing cultures of Geobacter sulfurreducens, a Geobacter species that cannot metabolize benzene. The phenol produced in G. metallireducens cultures was labeled with (18)O during growth in H2(18)O, as expected for anaerobic conversion of benzene to phenol. Analysis of whole-genome gene expression patterns indicated that genes for phenol metabolism were upregulated during growth on benzene but that genes for benzoate or toluene metabolism were not, further suggesting that phenol was an intermediate in benzene metabolism. Deletion of the genes for PpsA or PpcB, subunits of two enzymes specifically required for the metabolism of phenol, removed the capacity for benzene metabolism. These results demonstrate that benzene hydroxylation to phenol is an alternative to carboxylation for anaerobic benzene activation and suggest that this may be an important metabolic route for benzene removal in petroleum-contaminated groundwaters, in which Geobacter species are considered to play an important role in anaerobic benzene degradation.

  2. Survival of the North American strain of viral hemorrhagic septicemia virus (VHSV) in filtered seawater and seawater containing ovarian fluid, crude oil and serum-enriched culture medium

    Science.gov (United States)

    Kocan, R.M.; Hershberger, P.K.; Elder, N.E.

    2001-01-01

     The North American strain of viral hemorrhagic septicemia virus (NA-VHSV) could be recovered for up to 40 h in natural filtered seawater (27 ppt) with a 50% loss of infectivity after approximately 10 h at 15°C. Addition of 10 ppb North Slope crude oil to the seawater had no effect on virus survival. However, when various concentrations of teleost ovarian fluid were added to seawater, virus could be recovered after 72 h at 0.01% ovarian fluid and after 96 h at 1.0%. When cell culture medium supplemented with 10% fetal bovine serum was added to the seawater, 100% of the virus could be recovered for the first 15 d and 60% of the virus remained after 36 d. These findings quantify NA-VHSV infectivity in natural seawater and demonstrate that ovarian fluid, which occurs naturally during spawning events, significantly prolongs the survival and infectivity of the virus. The extended stabilization of virus in culture medium supplemented with serum allows for low titer field samples to be collected and transported in an unfrozen state without significant loss of virus titer.

  3. cultural

    Directory of Open Access Journals (Sweden)

    Irene Kreutz

    2006-01-01

    Full Text Available Es un estudio cualitativo que adoptó como referencial teorico-motodológico la antropología y la etnografía. Presenta las experiencias vivenciadas por mujeres de una comunidad en el proceso salud-enfermedad, con el objetivo de comprender los determinantes sócio-culturales e históricos de las prácticas de prevención y tratamiento adoptados por el grupo cultural por medio de la entrevista semi-estructurada. Los temas que emergieron fueron: la relación entre la alimentación y lo proceso salud-enfermedad, las relaciones con el sistema de salud oficial y el proceso salud-enfermedad y lo sobrenatural. Los dados revelaron que los moradores de la comunidad investigada tienen un modo particular de explicar sus procedimientos terapéuticos. Consideramos que es papel de los profesionales de la salud en sus prácticas, la adopción de abordajes o enfoques que consideren al individuo en su dimensión sócio-cultural e histórica, considerando la enorme diversidad cultural en nuestro país.

  4. Construction and screening of metagenomic libraries derived from enrichment cultures: generation of a gene bank for genes conferring alcohol oxidoreductase activity on Escherichia coli.

    Science.gov (United States)

    Knietsch, Anja; Waschkowitz, Tanja; Bowien, Susanne; Henne, Anke; Daniel, Rolf

    2003-03-01

    Enrichment of microorganisms with special traits and the construction of metagenomic libraries by direct cloning of environmental DNA have great potential for identifying genes and gene products for biotechnological purposes. We have combined these techniques to isolate novel genes conferring oxidation of short-chain (C(2) to C(4)) polyols or reduction of the corresponding carbonyls. In order to favor the growth of microorganisms containing the targeted genes, samples collected from four different environments were incubated in the presence of glycerol and 1,2-propanediol. Subsequently, the DNA was extracted from the four samples and used to construct complex plasmid libraries. Approximately 100,000 Escherichia coli strains of each library per test substrate were screened for the production of carbonyls from polyols on indicator agar. Twenty-four positive E. coli clones were obtained during the initial screen. Sixteen of them contained a plasmid (pAK101 to pAK116) which conferred a stable carbonyl-forming phenotype. Eight of the positive clones exhibited NAD(H)-dependent alcohol oxidoreductase activity with polyols or carbonyls as the substrates in crude extracts. Sequencing revealed that the inserts of pAK101 to pAK116 encoded 36 complete and 17 incomplete presumptive protein-encoding genes. Fifty of these genes showed similarity to sequenced genes from a broad collection of different microorganisms. The genes responsible for the carbonyl formation of E. coli were identified for nine of the plasmids (pAK101, pAK102, pAK105, pAK107 to pAK110, pAK115, and pAK116). Analyses of the amino acid sequences deduced from these genes revealed that three (orf12, orf14, and orf22) encoded novel alcohol dehydrogenases of different types, four (orf5, sucB, fdhD, and yabF) encoded novel putative oxidoreductases belonging to groups distinct from alcohol dehydrogenases, one (glpK) encoded a putative glycerol kinase, and one (orf1) encoded a protein which showed no similarity to any

  5. STUDY OF AEROBIC AND ANAEROBIC BACTERIA IN CHRONIC SUPPURATIVE OTITIS MEDIA

    Directory of Open Access Journals (Sweden)

    Satyanarayana

    2015-04-01

    Full Text Available One hundred six patients with clinical diagnosis of CSOM were investigated bacteriologically using appropriate aerobic and anaerobic techniques. Positive cultures were obtained in 100 specimens. Aerobic bacteria alone were present in 51%, anaerobic bacteria alone in 10% and a combination of both in 34% of aural swabs . The anaerobes isolated were Bacteroides 30%, Peptostreptococci 26%, followed by others. Aerobic bacteriology showed the predominance of Pseudomonas 30.4% followed by Staphylococcus aureus 21.4%. Metronidazole was found to be most effective (87% drug aga inst anaerobes and Amikacin (98% against aerobes.

  6. Development of a rapid lateral flow immunoassay test for detection of exosomes previously enriched from cell culture medium and body fluids

    Directory of Open Access Journals (Sweden)

    Myriam Oliveira-Rodríguez

    2016-08-01

    Full Text Available Exosomes are cell-secreted nanovesicles (40–200 nm that represent a rich source of novel biomarkers in the diagnosis and prognosis of certain diseases. Despite the increasingly recognized relevance of these vesicles as biomarkers, their detection has been limited due in part to current technical challenges in the rapid isolation and analysis of exosomes. The complexity of the development of analytical platforms relies on the heterogeneous composition of the exosome membrane. One of the most attractive tests is the inmunochromatographic strips, which allow rapid detection by unskilled operators. We have successfully developed a novel lateral flow immunoassay (LFIA for the detection of exosomes based on the use of tetraspanins as targets. We have applied this platform for the detection of exosomes purified from different sources: cell culture supernatants, human plasma and urine. As proof of concept, we explored the analytical potential of this LFIA platform to accurately quantify exosomes purified from a human metastatic melanoma cell line. The one-step assay can be completed in 15 min, with a limit of detection of 8.54×105 exosomes/µL when a blend of anti-CD9 and anti-CD81 were selected as capture antibodies and anti-CD63 labelled with gold nanoparticles as detection antibody. Based on our results, this platform could be well suited to be used as a rapid exosome quantification tool, with promising diagnostic applications, bearing in mind that the detection of exosomes from different sources may require adaptation of the analytical settings to their specific composition.

  7. Biodegradation potential of pure and mixed bacterial cultures for removal of 4-nitroaniline from textile dye wastewater.

    Science.gov (United States)

    Khalid, Azeem; Arshad, Muhammad; Crowley, David E

    2009-03-01

    Environmentally toxic aromatic amines including nitroanilines are commonly generated in dye contaminated wastewater in which azo dyes undergo degradation under anaerobic conditions. The aim of this study was to develop a process for biological treatment of 4-nitroaniline. Three bacteria identified as Acinetobacter sp., Citrobacter freundii and Klebsiella oxytoca were isolated from enrichment cultures of activated sludge on 4-nitroaniline, after which the isolates and the mixed culture were studied to determine optimal conditions for biodegradation. HPLC analyses showed the mixed culture was capable of complete removal of 100micromol/L of 4-nitroaniline within 72h under aerobic conditions. There was an inverse linear relationship (R(2)=0.96) between the rate of degradation (V) and 4-nitraoaniline concentrations [S] over 100-1000micromol/L. The bacterial culture was also capable of decolorizing structurally different azo dyes (Acid Red-88, Reactive Black-5, Direct Red-81, and Disperse Orange-3) and also degraded nitrobenzene. Our findings show that enrichment cultures from activated sludge can be effective for the removal of dyes and their toxic intermediates, and that treatment may best be accomplished using an anaerobic-aerobic process.

  8. New perspectives in anaerobic digestion

    DEFF Research Database (Denmark)

    van Lier, J.B.; Tilche, A.; Ahring, Birgitte Kiær;

    2001-01-01

    The IWA specialised group on anaerobic digestion (AD) is one of the oldest working groups of the former IAWQ organisation. Despite the fact that anaerobic technology dates back more than 100 years, the technology is still under development, adapting novel treatment systems to the modern...... requirements. In fact, most advances were achieved during the last three decades, when high-rate reactor systems were developed and a profound insight was obtained in the microbiology of the anaerobic communities. This insight led to a better understanding of anaerobic treatment and, subsequently, to a broader...

  9. Enrichment of Breast Cancer Stem Cells by Serum-free Shaking Suspension Culture%无血清摇动悬浮培养富集乳腺癌干细胞

    Institute of Scientific and Technical Information of China (English)

    闫文星; 陈玉丙; 张红梅; 国春龙; 王铁君

    2011-01-01

    Objective To investigate the characteristics of division of breast cancer MCF-7 cells under various conditions so as to develop a rapid and effective method for enrichment of breast cancer stem cells. Methods MCF-7 cells were subjected to static culture in complete medium (group A), shaking culture in complete medium (group B), static culture with cytokine (group C) and shaking culture with cytokine (group D) respectively, for 12, 24 and 36 h, then observed for division under inverted phase contrast microscope, based on which clone formation rate was calculated, and determined for the percentage of CD44+/CD24"/low lymphocyte subsets. Results In groups B and C, rod-like divisions were observed in both about 30% of cells 12 h, and in about 50% and about 60% of cells respectively 24 h, while large cell clones were formed 36 h after culture. However, in group D, rod-like division was observed in about 50% of cells 12 h, and in about 80% of cells 24 h when several cell clones appeared, while the number of cell clones decreased 36 h after culture. The percentage of CD44+/CD24"/km lymphocyte subsets 12 h after culture in group D (8. 05%) was 8 times of those in group B (0. 99%) and 2. 1 times of those in group C (3. 80%). However, the percentage 24 h after culture in group D (15. 24%) was 3 times of those in group B (4. 83%) and 6 times of those in group C (2. 30%). The percentage in group D decreased to 9. 68% 36 h after culture, which was about 9 times of those in groups B (0. 95%) and C (1. 03%). Conclusion The mitotic division of MCF-7 cells was accelerated in shaking culture with cytokine, while the percentage of CD44+/CD24'/low lymphocyte subsets increased rapidly, and stem cell pool increased significantly, indicating serum-free shaking suspension culture a rapid and effective method for enrichment of breast cancer stem cells.%目的 探讨不同培养条件下乳腺癌MCF-7细胞的分裂特点,建立快速、有效的乳腺癌干细胞富集方法.方法 将MCF-7

  10. Anaerobic biodegradation of lipids of the marine microalga Nannochloropsis salina

    NARCIS (Netherlands)

    Sinninghe Damsté, J.S.; Grossi, V.; Blokker, P.

    2001-01-01

    In order to determine the susceptibility to anaerobic biodegradation of the different lipid biomarkers present in a marine microalga containing algaenan, portions of one large batch of cultured Nannochloropsis salina (Eustigmatophyceae) were incubated in anoxic sediment slurries for various times. A

  11. Microbial degradation of 4-monobrominated diphenyl ether with anaerobic sludge

    Energy Technology Data Exchange (ETDEWEB)

    Shih, Yang-hsin, E-mail: yhs@ntu.edu.tw [Department of Agricultural Chemistry, National Taiwan University, Taipei 106, Taiwan, ROC (China); Chou, Hsi-Ling [Department of Soil and Environmental Sciences, National Chung Hsing University, Taichung 407, Taiwan, ROC (China); Peng, Yu-Huei [Department of Agricultural Chemistry, National Taiwan University, Taipei 106, Taiwan, ROC (China)

    2012-04-30

    Highlights: Black-Right-Pointing-Pointer BDE-3 was degraded with two anaerobes in different rates. Black-Right-Pointing-Pointer Glucose addition augment the debromination efficiencies. Black-Right-Pointing-Pointer Hydrogen gas was detected and relative microbes were identified. Black-Right-Pointing-Pointer Extra-carbon source enhanced degradation partial due to H{sub 2}-generation bacteria. - Abstract: Polybrominated diphenyl ethers (PBDEs) are widely used flame retardant additives for many plastic and electronic products. Owing to their ubiquitous distribution in the environment, multiple toxicity to humans, and increasing accumulation in the environment, the fate of PBDEs is of serious concern for public safety. In this study, the degradation of 4-monobrominated diphenyl ether (BDE-3) in anaerobic sludge and the effect of carbon source addition were investigated. BDE-3 can be degraded by two different anaerobic sludge samples. The by-products, diphenyl ether (DE) and bromide ions, were monitored, indicating the reaction of debromination within these anaerobic samples. Co-metabolism with glucose facilitated BDE-3 biodegradation in terms of kinetics and efficiency in the Jhongsing sludge. Through the pattern of amplified 16S rRNA gene fragments in denatured gradient gel electrophoresis (DGGE), the composition of the microbial community was analyzed. Most of the predominant microbes were novel species. The fragments enriched in BDE-3-degrading anaerobic sludge samples are presumably Clostridium sp. This enrichment coincides with the H{sub 2} gas generation and the facilitation of debromination during the degradation process. Findings of this study provide better understanding of the biodegradation of brominated DEs and can facilitate the prediction of the fate of PBDEs in the environment.

  12. Cultivation and irradiation of human fibroblasts in a medium enriched with platelet lysate for obtaining feeder layer in epidermal cell culture; Cultivo e irradiacao de fibroblastos humanos em meio enriquecido com lisado de plaquetas para obtencao de camada de sustentacao em culturas de celulas da epiderme

    Energy Technology Data Exchange (ETDEWEB)

    Yoshito, Daniele

    2011-07-01

    For over 30 years, the use of culture medium, enriched with bovine serum, and murines fibroblasts, with the rate of proliferation controlled by irradiation or by share anticarcinogenic drugs, has been playing successfully its role in assisting in the development of keratinocytes in culture, for clinical purposes. However, currently there is a growing concern about the possibility of transmitting prions and animals viruses to transplanted patients. Taking into account this concern, the present work aims to cultivate human fibroblasts in a medium enriched with human platelets lysate and determine the irradiation dose of these cells, for obtaining feeder layer in epidermal cell culture. For carrying out the proposed objective, platelets lysis has standardized, this lysate was used for human fibroblasts cultivation and the irradiation dose enough to inhibit its duplication was evaluated. Human keratinocytes were cultivated in these feeder layers, in culture medium enriched with the lysate. With these results we conclude that the 10% platelets lysate promoted a better adhesion and proliferation of human fibroblasts and in all dose levels tested (60 to 300 Gy), these had their mitotic activity inactivated by ionizing irradiation, being that the feeder layers obtained with doses from 70 to 150 Gy were those that provided the best development of keratinocytes in medium containing 2.5% of human platelet lysate. Therefore, it was possible to standardize both the cultivation of human fibroblasts as its inactivation for use as feeder layer in culture of keratinocytes, so as to eliminate xenobiotics components. (author)

  13. Anaerobic Digestion of Piggery Waste

    NARCIS (Netherlands)

    Velsen, van A.F.M.

    1981-01-01

    Anaerobic digestion is a biological process by which organic matter is converted to methane and carbon dioxide by microbes in the absence of air (oxygen). In nature, anaerobic conversions occur at all places where organic material accumulates and the supply of oxygen is deficient, e.g. in marshes an

  14. Economic viability of anaerobic digestion

    Energy Technology Data Exchange (ETDEWEB)

    Wellinger, A. [INFOENERGIE, Ettenhausen (Switzerland)

    1996-01-01

    The industrial application of anaerobic digestion is a relatively new, yet proven waste treatment technology. Anaerobic digestion reduces and upgrades organic waste, and is a good way to control air pollution as it reduces methane and nitrous gas emissions. For environmental and energy considerations, anaerobic digestion is a nearly perfect waste treatment process. However, its economic viability is still in question. A number of parameters - type of waste (solid or liquid), digester system, facility size, product quality and end use, environmental requirements, cost of alternative treatments (including labor), and interest rates - define the investment and operating costs of an anaerobic digestion facility. Therefore, identical facilities that treat the same amount and type of waste may, depending on location, legislation, and end product characteristics, reveal radically different costs. A good approach for evaluating the economics of anaerobic digestion is to compare it to treatment techniques such as aeration or conventional sewage treatment (for industrial wastewater), or composting and incineration (for solid organic waste). For example, the cost (per ton of waste) of in-vessel composting with biofilters is somewhat higher than that of anaerobic digestion, but the investment costs 1 1/2 to 2 times more than either composting or anaerobic digestion. Two distinct advantages of anaerobic digestion are: (1) it requires less land than either composting or incinerating, which translates into lower costs and milder environmental and community impacts (especially in densely populated areas); and (2) it produces net energy, which can be used to operate the facility or sold to nearby industries.

  15. Anaerobic digestion of solid material

    DEFF Research Database (Denmark)

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

    2007-01-01

    A new multidimensional (3 and 2D) anaerobic digestion model for cylindrical reactor with non-uniform influent concentration distributions was developed to study the way in which mixing intensity affects the efficiency of continuous-flow anaerobic digestion. Batch experiments reported and simulate...

  16. Gravimetric enrichment of high lipid and starch accumulating microalgae.

    Science.gov (United States)

    Hassanpour, Morteza; Abbasabadi, Mahsa; Ebrahimi, Sirous; Hosseini, Maryam; Sheikhbaglou, Ahmad

    2015-11-01

    This study presents gravimetric enrichment of mixed culture to screen starch and lipid producing species separately in a sequencing batch reactor. In the enriched starch-producing mixed culture photobioreactor, the starch content at the end of steady state batch became 3.42 times the beginning of depletion. Whereas in the enriched lipid-producing photobioreactor, the lipid content at the end of steady state batch became 3 times the beginning of famine phase. The obtained results revealed that the gravimetric enrichment is a suitable screening method for specific production of storage compounds in none-sterile large-scaled condition.

  17. In vitro Characterization of Phenylacetate Decarboxylase, a Novel Enzyme Catalyzing Toluene Biosynthesis in an Anaerobic Microbial Community.

    Science.gov (United States)

    Zargar, K; Saville, R; Phelan, R M; Tringe, S G; Petzold, C J; Keasling, J D; Beller, H R

    2016-08-10

    Anaerobic bacterial biosynthesis of toluene from phenylacetate was reported more than two decades ago, but the biochemistry underlying this novel metabolism has never been elucidated. Here we report results of in vitro characterization studies of a novel phenylacetate decarboxylase from an anaerobic, sewage-derived enrichment culture that quantitatively produces toluene from phenylacetate; complementary metagenomic and metaproteomic analyses are also presented. Among the noteworthy findings is that this enzyme is not the well-characterized clostridial p-hydroxyphenylacetate decarboxylase (CsdBC). However, the toluene synthase under study appears to be able to catalyze both phenylacetate and p-hydroxyphenylacetate decarboxylation. Observations suggesting that phenylacetate and p-hydroxyphenylacetate decarboxylation in complex cell-free extracts were catalyzed by the same enzyme include the following: (i) the specific activity for both substrates was comparable in cell-free extracts, (ii) the two activities displayed identical behavior during chromatographic separation of cell-free extracts, (iii) both activities were irreversibly inactivated upon exposure to O2, and (iv) both activities were similarly inhibited by an amide analog of p-hydroxyphenylacetate. Based upon these and other data, we hypothesize that the toluene synthase reaction involves a glycyl radical decarboxylase. This first-time study of the phenylacetate decarboxylase reaction constitutes an important step in understanding and ultimately harnessing it for making bio-based toluene.

  18. Bio-oil upgrading strategies to improve PHA production from selected aerobic mixed cultures.

    Science.gov (United States)

    Moita Fidalgo, Rita; Ortigueira, Joana; Freches, André; Pelica, João; Gonçalves, Magarida; Mendes, Benilde; Lemos, Paulo C

    2014-06-25

    Recent research on polyhydroxyalkanoates (PHAs) has focused on developing cost-effective production processes using low-value or industrial waste/surplus as substrate. One of such substrates is the liquid fraction resulting from pyrolysis processes, bio-oil. In this study, valorisation of bio-oil through PHA production was investigated. The impact of the complex bio-oil matrix on PHA production by an enriched mixed culture was examined. The performance of the direct utilization of pure bio-oil was compared with the utilization of three defined substrates contained in this bio-oil: acetate, glucose and xylose. When compared with acetate, bio-oil revealed lower capacity for polymer production as a result of a lower polymer yield on substrate and a lower PHA cell content. Two strategies for bio-oil upgrade were performed, anaerobic fermentation and vacuum distillation, and the resulting liquid streams were tested for polymer production. The first one was enriched in volatile fatty acids and the second one mainly on phenolic and long-chain fatty acids. PHA accumulation assays using the upgraded bio-oils attained polymer yields on substrate similar or higher than the one achieved with acetate, although with a lower PHA content. The capacity to use the enriched fractions for polymer production has yet to be optimized. The anaerobic digestion of bio-oil could also open-up the possibility to use the fermented bio-oil directly in the enrichment process of the mixed culture. This would increase the selective pressure toward an optimized PHA accumulating culture selection.

  19. Impact of salinity on the anaerobic metabolism of phosphate-accumulating organisms (PAO) and glycogen-accumulating organisms (GAO).

    Science.gov (United States)

    Welles, L; Lopez-Vazquez, C M; Hooijmans, C M; van Loosdrecht, M C M; Brdjanovic, D

    2014-09-01

    The use of saline water as secondary quality water in urban environments for sanitation is a promising alternative towards mitigating fresh water scarcity. However, this alternative will increase the salinity in the wastewater generated that may affect the biological wastewater treatment processes, such as biological phosphorus removal. In addition to the production of saline wastewater by the direct use of saline water in urban environments, saline wastewater is also generated by some industries. Intrusion of saline water into the sewers is another source of salinity entering the wastewater treatment plant. In this study, the short-term effects of salinity on the anaerobic metabolism of phosphate-accumulating organisms (PAO) and glycogen-accumulating organisms (GAO) were investigated to assess the impact of salinity on enhanced biological phosphorus removal. Hereto, PAO and GAO cultures enriched at a relatively low salinity level (0.02 % W/V) were exposed to salinity concentrations of up to 6 % (as NaCl) in anaerobic batch tests. It was demonstrated that both PAO and GAO are affected by higher salinity levels, with PAO being the more sensitive organisms to the increasing salinity. The maximum acetate uptake rate of PAO decreased by 71 % when the salinity increased from 0 to 1 %, while that of GAO decreased by 41 % for the same salinity increase. Regarding the stoichiometry of PAO, a decrease in the P-release/HAc uptake ratio accompanied with an increase in the glycogen consumption/HAc uptake ratio was observed for PAO when the salinity increased from 0 to 2 % salinity, indicating a metabolic shift from a poly-P-dependent to a glycogen-dependent metabolism. The anaerobic maintenance requirements of PAO and GAO increased as the salinity concentrations risen up to 4 % salinity.

  20. 应用渗透膜高密度培养富硒螺旋藻%Application of Osmotic Membrane for High Cell Density Culture of Selenium-Enriched Spirulina platensis

    Institute of Scientific and Technical Information of China (English)

    任璐艳; 靳兴媛; 周永林; 张逸波; 凌钦婕; 黄峙

    2012-01-01

    应用透析袋(OT法)组装光生物反应器,与鼓泡式生物反应器(AB法)和摇瓶(SB法)进行对比实验,探讨利用渗透膜建立高密度培养富硒螺旋藻(Se-sP)方法的可行性。称质量法监测并推算藻细胞生长率和最大生物量,荧光光度法测定se-sP及其培养液中硒含量和形态变化,分光光度法检测Se-SP中总蛋白(TP)、藻蓝蛋白(PC)、叶绿素a(Chla)、总胡萝卜素(Caro)、水溶性多糖(SPS)及巯基(-SH)含量等主要营养构成,培养液中无机碳(IC)和有机溶解碳(DOC)的含量采用自动分析仪测定。结果发现:OT法培养Se-SP的最大生物量可达9.6·g/L,是AB和SB方法培养的3-5倍,其中有机硒转化率及TP、PC、Chla、SPS和-SH等主要营养成分的含量均有明显提高。相应地,培养基中IC剩余减少,DOC无明显累积。结果提示,OT法可实现Se-SP高密度培养,并有望获得优质Se-SP产品,其优势体现在高细胞增殖速率、高有机硒转化率、高无机营养源消耗率和低有机碳的积累,便于培养液的再生利用,减少废液污染。%Selenium-enriched Spirulinaplatensis (Se-SP) was cultured in osmotic membrane tube (OT) and compared with air blowing bioreactor (AB) and shaking bottle (SB) cultures under the same conditions. Se supplementation was 100 μ g/mL in the form of sodium selenite. Growth rate and maximum biomass were monitored and deduced according to dry weight at designated time points. Total Se, organic Se and inorganic Se in Se-SP as well as soluble and insoluble Se species in culture medium were determined using a fluorimeter. Total protein (TP), phycocyanin (PC), chlorophyll a (Chla), carotenoids (Caro), total polysac- charides (SPS) and sulfydryl groups (-SH) in Se-SP were also detected colorimetrically. The contents of inorganic carbon (IC) and dissolved organic carbon

  1. Isolation and Cultivation of Anaerobes

    DEFF Research Database (Denmark)

    Aragao Börner, Rosa

    2016-01-01

    Anaerobic microorganisms play important roles in different biotechnological processes. Their complex metabolism and special cultivation requirements have led to less isolated representatives in comparison to their aerobic counterparts.In view of that, the isolation and cultivation of anaerobic...... microorganisms is still a promising venture, and conventional methodologies as well as considerations and modifications are presented here. An insight into new methodologies and devices as well as a discussion on future perspectives for the cultivation of anaerobes may open the prospects of the exploitation...

  2. Microbial communities mediating algal detritus turnover under anaerobic conditions

    Directory of Open Access Journals (Sweden)

    Jessica M. Morrison

    2017-01-01

    Full Text Available Background Algae encompass a wide array of photosynthetic organisms that are ubiquitously distributed in aquatic and terrestrial habitats. Algal species often bloom in aquatic ecosystems, providing a significant autochthonous carbon input to the deeper anoxic layers in stratified water bodies. In addition, various algal species have been touted as promising candidates for anaerobic biogas production from biomass. Surprisingly, in spite of its ecological and economic relevance, the microbial community involved in algal detritus turnover under anaerobic conditions remains largely unexplored. Results Here, we characterized the microbial communities mediating the degradation of Chlorella vulgaris (Chlorophyta, Chara sp. strain IWP1 (Charophyceae, and kelp Ascophyllum nodosum (phylum Phaeophyceae, using sediments from an anaerobic spring (Zodlteone spring, OK; ZDT, sludge from a secondary digester in a local wastewater treatment plant (Stillwater, OK; WWT, and deeper anoxic layers from a seasonally stratified lake (Grand Lake O’ the Cherokees, OK; GL as inoculum sources. Within all enrichments, the majority of algal biomass was metabolized within 13–16 weeks, and the process was accompanied by an increase in cell numbers and a decrease in community diversity. Community surveys based on the V4 region of the 16S rRNA gene identified different lineages belonging to the phyla Bacteroidetes, Proteobacteria (alpha, delta, gamma, and epsilon classes, Spirochaetes, and Firmicutes that were selectively abundant under various substrate and inoculum conditions. Within all kelp enrichments, the microbial communities structures at the conclusion of the experiment were highly similar regardless of the enrichment source, and were dominated by the genus Clostridium, or family Veillonellaceae within the Firmicutes. In all other enrichments the final microbial community was dependent on the inoculum source, rather than the type of algae utilized as substrate

  3. Microbial communities mediating algal detritus turnover under anaerobic conditions

    Science.gov (United States)

    Morrison, Jessica M.; Murphy, Chelsea L.; Baker, Kristina; Zamor, Richard M.; Nikolai, Steve J.; Wilder, Shawn; Elshahed, Mostafa S.

    2017-01-01

    Background Algae encompass a wide array of photosynthetic organisms that are ubiquitously distributed in aquatic and terrestrial habitats. Algal species often bloom in aquatic ecosystems, providing a significant autochthonous carbon input to the deeper anoxic layers in stratified water bodies. In addition, various algal species have been touted as promising candidates for anaerobic biogas production from biomass. Surprisingly, in spite of its ecological and economic relevance, the microbial community involved in algal detritus turnover under anaerobic conditions remains largely unexplored. Results Here, we characterized the microbial communities mediating the degradation of Chlorella vulgaris (Chlorophyta), Chara sp. strain IWP1 (Charophyceae), and kelp Ascophyllum nodosum (phylum Phaeophyceae), using sediments from an anaerobic spring (Zodlteone spring, OK; ZDT), sludge from a secondary digester in a local wastewater treatment plant (Stillwater, OK; WWT), and deeper anoxic layers from a seasonally stratified lake (Grand Lake O’ the Cherokees, OK; GL) as inoculum sources. Within all enrichments, the majority of algal biomass was metabolized within 13–16 weeks, and the process was accompanied by an increase in cell numbers and a decrease in community diversity. Community surveys based on the V4 region of the 16S rRNA gene identified different lineages belonging to the phyla Bacteroidetes, Proteobacteria (alpha, delta, gamma, and epsilon classes), Spirochaetes, and Firmicutes that were selectively abundant under various substrate and inoculum conditions. Within all kelp enrichments, the microbial communities structures at the conclusion of the experiment were highly similar regardless of the enrichment source, and were dominated by the genus Clostridium, or family Veillonellaceae within the Firmicutes. In all other enrichments the final microbial community was dependent on the inoculum source, rather than the type of algae utilized as substrate. Lineages enriched

  4. Two-Dimensional Stable Isotope Fractionation During Aerobic and Anaerobic Alkane Biodegradation and Implications for the Field

    Science.gov (United States)

    El Morris, Brandon; Suflita, Joseph M.; Richnow, Hans-Hermann

    2010-05-01

    Quantitatively, n-alkanes comprise a major portion of most crude oils. In petroliferous formations, it may be possible to relate the loss of these compounds to the levels of biodegradation occurring in situ [1]. Moreover, it is important to develop indicators of alkane degradation that may be used to monitor bioremediation of hydrocarbon-impacted environments. Desulfoglaeba alkanexedens and Pseudomonas putida GPo1 were used to determine if carbon and hydrogen stable isotope fractionation could differentiate between n-alkane degradation under anaerobic and aerobic conditions, respectively in the context of the Rayleigh equation model [2]. Bacterial cultures were sacrificed by acidification and headspace samples were analyzed for stable isotope composition using gas chromatography-isotope ratio mass spectrometry. Carbon enrichment factors (bulk) for anaerobic and aerobic biodegradation of hexane were -5.52 ± 0.2‰ and -4.34 ± 0.3‰, respectively. Hydrogen enrichment during hexane degradation was -43.14 ± 6.32‰ under sulfate-reducing conditions, and was too low for quantification during aerobiosis. Collectively, this indicates that the correlation between carbon and hydrogen stable isotope fractionation (may be used to help elucidate in situ microbial processes in oil reservoirs, and during intrinsic as well as engineered remediation efforts. References 1. Asif, M.; Grice, K.; Fazeelat, T., Assessment of petroleum biodegradation using stable hydrogen isotopes of individual saturated hydrocarbon and polycyclic aromatic hydrocarbon distributions in oils from the Upper Indus Basin, Pakistan. Organic Geochemistry 2009, 40, (3), 301-311. 2. Fischer, A.; Herklotz, I.; Herrmann, S.; Thullner, M.; Weelink, S. A. B.; Stams, A., J. M.; Schloemann, M.; Richnow, H.-H.; Vogt, C., Combined carbon and hydrogen isotope fractionation investigations for elucidating benzene biodegradation pathways. Environ. Sci. Technol. 2008, 42, 4356-4363.

  5. A review of anaerobic treatment of saline wastewater.

    Science.gov (United States)

    Xiao, Yeyuan; Roberts, Deborah J

    2010-01-01

    Large volumes of saline (> 2% w/v NaCl) wastewaters are discharged from many industries; e.g. seafood processing, textile dyeing, oil and gas production, tanneries and drinking water treatment processes. Although anaerobic treatment would be the most cost-effective and sustainable technology for the treatment of many of these saline wastewaters, the salinity is considered to be inhibitory to anaerobic biological treatment processes. The recent applications of salt-tolerant cultures for the treatment of wastewaters from seafood processing and ion-exchange processes suggest that biological systems can be used to treat salty wastewaters. Additionally, organisms capable of anaerobic degradation of contaminants in saline solutions have been observed in marine sediments and have been characterized during the last two decades. This manuscript provides a review of the recent research on anaerobic treatment of saline wastewater and bacterial consortia capable of the anaerobic degradation of pollutants in saline solutions, documenting that the biological treatment of saline wastewaters is promising.

  6. Decrease in anaerobe-related bacteraemias and increase in Bacteroides species isolation rate from 1998 to 2007: a retrospective study.

    Science.gov (United States)

    Lazarovitch, Tsilia; Freimann, Sarit; Shapira, Galina; Blank, Helena

    2010-06-01

    Conflicting data have accumulated in recent years regarding the incidence of anaerobic bacteraemias. The aim of this study was to determine the prevalence of bacteraemias due to anaerobic bacteria and evaluate the importance of anaerobic blood cultures in a university hospital in Israel. A retrospective survey which focused on anaerobic blood culture bottles was performed on blood cultures received in our laboratory during the decade from January 1998 to December 2007. Anaerobic-related bacteraemias decreased during that period, whereas a significant increase was observed in Bacteroides species isolated from the blood cultures (from 18% during 1998-2002 to 43% during 2003-2007). Comparison of the medical records of 54 patients with Bacteroides-related bacteraemia during the two end periods (1998-1999 and 2006-2007) revealed a marked increase in complex underlying diseases. Hypertension and diabetes mellitus type II were found in 29% of the patients in 1998-1999 and increased to 43-45% of the patients in 2006-2007. Ischemic heart disease also increased from 14% of the patients in 1998-1999 to 43% in 2006-2007. We conclude that although positive anaerobic blood cultures account for a small percentage of positive blood samples, the growing involvement of Bacteroides species-related bacteraemias together with an increase in complex underlying diseases in these patients emphasize the importance of anaerobic blood cultures, particularly in patients with co-morbidities.

  7. Bioaugmentation with an acetate-oxidising consortium as a tool to tackle ammonia inhibition of anaerobic digestion

    DEFF Research Database (Denmark)

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

    2013-01-01

    Ammonia is the major inhibitor of anaerobic digestion (AD) process in biogas plants. In the current study, the bioaugmentation of the ammonia tolerant SAO co-culture (i.e. Clostridium ultunense spp. nov. in association with Methanoculleus spp. strain MAB1) in a mesophilic up-flow anaerobic sludge...

  8. Combined anaerobic ammonium and methane oxidation for nitrogen and methane removal.

    Science.gov (United States)

    Zhu, Baoli; Sánchez, Jaime; van Alen, Theo A; Sanabria, Janeth; Jetten, Mike S M; Ettwig, Katharina F; Kartal, Boran

    2011-12-01

    Anammox (anaerobic ammonium oxidation) is an environment-friendly and cost-efficient nitrogen-removal process currently applied to high-ammonium-loaded wastewaters such as anaerobic digester effluents. In these wastewaters, dissolved methane is also present and should be removed to prevent greenhouse gas emissions into the environment. Potentially, another recently discovered microbial pathway, n-damo (nitrite-dependent anaerobic methane oxidation) could be used for this purpose. In the present paper, we explore the feasibility of simultaneously removing methane and ammonium anaerobically, starting with granules from a full-scale anammox bioreactor. We describe the development of a co-culture of anammox and n-damo bacteria using a medium containing methane, ammonium and nitrite. The results are discussed in the context of other recent studies on the application of anaerobic methane- and ammonia-oxidizing bacteria for wastewater treatment.

  9. Anaerobic Biodegradation of Detergent Surfactants

    OpenAIRE

    Erich Jelen; Ute Merrettig-Bruns

    2009-01-01

    Detergent surfactantscan 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 str...

  10. Performance and microbial community analysis of the anaerobic reactor with coke oven gas biomethanation and in situ biogas upgrading

    DEFF Research Database (Denmark)

    Wang, Wen; Xie, Li; Luo, Gang;

    2013-01-01

    (HFM). With pH control at 8.0, the added H2 and CO were fully consumed and no negative effects on the anaerobic degradation of sewage sludge were observed. The maximum CH4 content in the biogas was 99%. The addition of SCOG resulted in enrichment and dominance of homoacetogenetic genus Treponema......A new method for simultaneous coke oven gas (COG) biomethanation and in situ biogas upgrading in anaerobic reactor was developed in this study. The simulated coke oven gas (SCOG) (92% H2 and 8% CO) was injected directly into the anaerobic reactor treating sewage sludge through hollow fiber membrane...

  11. The role of anaerobic digestion in the emerging energy economy.

    Science.gov (United States)

    Batstone, Damien John; Virdis, Bernardino

    2014-06-01

    Anaerobic digestion is the default process for biological conversion of residue organics to renewable energy and biofuel in the form of methane. However, its scope of application is expanding, due to availability of new technologies, and the emerging drivers of energy and nutrient conservation and recovery. Here, we outline two of these new application areas, namely wastewater nutrient and energy recovery, and generation of value added chemicals through mixed culture biotechnology. There exist two options for nutrient and energy recovery from domestic wastewater: low energy mainline and partition-release-recovery. Both are heavily dependent on anaerobic digestion as an energy generating and nutrient release step, and have been enabled by new technologies such as low emission anaerobic membrane processes. The area of mixed culture biotechnology has been previously identified as a key industrial opportunity, but is now moving closer to application due application of existing and new technologies. As well as acting as a core technology option in bioproduction, anaerobic digestion has a key role in residual waste valorization and generation of energy for downstream processing. These new application areas and technologies are emerging simultaneously with substantial advances in knowledge of underlying mechanisms such as electron transfer, understanding of which is critical to development of the new application areas.

  12. Potential application of anaerobic extremophiles for hydrogen production

    Science.gov (United States)

    Pikuta, Elena V.; Hoover, Richard B.

    2004-11-01

    In processes of the substrate fermentation most anaerobes produce molecular hydrogen as a waste end product, which often controls the culture growth as an inhibitor. Usually in nature the hydrogen is easily removed from an ecosystem, due to its physical features, and an immediate consumption by the secondary anaerobes that sometimes behave as competitors for electron donors; a classical example of this kind of substrate competition in anaerobic microbial communities is the interaction between methanogens and sulfate- or sulfur-reducers. Previously, on the mixed cultures of anaerobes at neutral pH, it was demonstrated that bacterial hydrogen production could provide a good alternative energy source. At neutral pH the original cultures could easily contaminated by methanogens, and the 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 furthermore, the cultivation with pathogenic contaminants on an industrial scale would create an unsafe situation. In our laboratory the experiments with obligately alkaliphilic bacteria producing hydrogen as an end metabolic product were performed at different conditions. The mesophilic, haloalkaliphilic and obligately anaerobic bacterium Spirochaeta americana ASpG1T was studied and various cultivation regimes were compared for the most effective hydrogen production. In a highly mineralized media with pH 9.5-10.0 not many known methanogens are capable of growth, and the probability of developing pathogenic contaminants is theoretically is close to zero (in medicine carbonate- saturated solutions are applied as antiseptics). Therefore the cultivation of alkaliphilic hydrogen producing bacteria could be considered as a safe and economical process for large-scale industrial bio-hydrogen production in the future. Here we present and discuss the experimental data

  13. Effect of temperature and organic nutrients on the biodegradation of linear alkylbenzene sulfonate (LAS) during the composting of anaerobically digested sludge from a wastewater treatment plant.

    Science.gov (United States)

    Sanz, E; Prats, D; Rodríguez, M; Camacho, A

    2006-01-01

    Limits on the application of biosolids (anaerobically processed sludges from wastewater treatment plants) as fertilizers for the amendment of soil are becoming greater because of the accumulation of recalcitrant substances, making necessary the use of techniques that bring the concentration of xenobiotics to lower concentrations than those permitted. In general, the biosolids composting process is sufficient to reduce the usual concentration of linear alkylbenzene sulfonates (LAS) to low levels. In this work, an assessment is made on the effect of temperature in the capacity of enriched bacterial populations to biodegrade LAS, together with the influence that the available nutrients may have in the biodegradation of these compounds. The results show that the microbial metabolism of LAS was not observed in the thermophilic range. The optimum temperature for the biodegradation of LAS appears to be around 40 degrees C, this is, the lowest assayed here, and at this temperature the differences in the biodegradation of LAS among the nutritionally supplemented cultures are small.

  14. The effect of cyclic anaerobic-aerobic conditions on biodegradation of azo dyes.

    Science.gov (United States)

    Yaşar, Semra; Cirik, Kevser; Cinar, Ozer

    2012-03-01

    The effect of cyclic anaerobic-aerobic conditions on the biodegradative capability of the mixed microbial culture for the azo dye Remazol Brilliant Violet 5R (RBV-5R) was investigated in the sequencing batch reactor (SBR) fed with a synthetic textile wastewater. The SBR had a 12-h cycle time with anaerobic-aerobic periods of 3/9, 6/6 and 9/3 h. General SBR performance was assessed by measurement of catabolic enzymes (catechol 2,3-dioxygenase, azo reductase), chemical oxygen demand (COD), color and amount of aromatic amines. In this study, under steady-state conditions, the anaerobic period of the cyclic SBR was found to allow the reductive decolorization of azo dye. Longer anaerobic periods resulted in higher color removal efficiencies, approximately 71% for the 3-h, 87% for 6-h and 92% for the 9-h duration. Total COD removal efficiencies were over 84% under each of the cyclic conditions and increased as the length of the anaerobic period was increased; however, the highest color removal rate was attained for the cycle with the shortest anaerobic period of 3 h. During the decolorization of RBV-5R, two sulfonated aromatic amines (benzene based and naphthalene based) were formed. Additionally, anaerobic azo reductase enzyme was found to be positively affected with the increasing duration of the anaerobic period; however; it was vice versa for the aerobic catechol 2,3-dioxygenase (C23DO) enzyme.

  15. New techniques for growing anaerobic bacteria: Experiments with Clostridium butyricum and Clostridium acetobutylicum

    Energy Technology Data Exchange (ETDEWEB)

    Adler, H.I.; Crow, W.D.; Hadden, C.T.; Hall, J.; Machanoff, R.

    1983-01-01

    Stable membrane fragments derived from Escherichia coli produce and maintain strict anaerobic conditions when added to liquid or solid bacteriological media. Techniques for growing Clostridium butyricum and Clostridium acetobutylicum in membrane containing media are described. Liquid cultures initiated by very small inocula can be grown in direct contact with air. In solid media, colonies develop rapidly from individual cells even without incubation in anaerobic jars or similar devices. Observations on growth rates, spontaneous mutations, radiation and oxygen sensitivity of anaerobic bacteria have been made using these new techniques.

  16. 水培条件下几种观赏植物对铅的富集特征%Enrichment Characteristics of Pb by Several Kinds of Ornamental Plants Under Hydroponic Culture

    Institute of Scientific and Technical Information of China (English)

    李翠兰; 邵泽强; 王玉军; 张晋京

    2011-01-01

    通过水培试验,研究了铅处理质量浓度为0~200 mg·L-1条件下,7种观赏植物(包括四季海棠、百日草、茶花凤仙、金鱼草、金盏菊、天竺葵和雁来红)的生长反应和富集特征.结果表明:供试观赏植物在试验的铅胁迫水平下都具有较强的耐性;四季海棠和百日草的地上部最大铅质量分数分别达到了1 229.2、1 209.7 mg·kg-1,满足超富集植物的临界质量分数标准,同时它们在试验铅处理质量浓度下的地上部铅质量分数和富集系数均显著高于其他观赏植物;四季海棠和百日草的铅转移系数最高值可达到0.9以上,并且百日草铅质量浓度在200 mg·L-1时的转移系数仍高于100 mg·L-1时的转移系数.%The growth response and enrichment characteristics of seven ornamental plants, Begonia semperflorens, Zinnia elegan, Impatiens balsamina, Antirrhinum majus, Calendula officinalis, Pelargonium hortorum and Amaranthus tricolo, were studied under hydroponic culture with 0-200 mg · L-1 of Pb. Results showed that all the tested ornamental plants had higher tolerance to Pb in the range of experimental Pb stress levels. The highest Pb mass fractions in the above-ground parts of B. semperflorens and Z. elegan were 1 229.2 and 1 209.7 mg · kg-1, respectively, which reached the critical standard of Pb hyperaccumulating plants. And the Pb mass fraction and bioaccumulation coefficient in the above-ground parts of B. semperflorens and Z. elegan was significantly higher than those of the other five ornamental plants. The highest Pb translocation factors for B. semperflorens and Z. elegan were over 0.9, and the Pb translocation factor for Z. elegan was still higher under 200 mg · L-1 than under 100 mg · L-1 of Pb.

  17. Microbiology, ecology, and application of the nitrite-dependent anaerobic methane oxidation process

    OpenAIRE

    Li-Dong eShen; Bao-lan eHu

    2012-01-01

    Nitrite-dependent anaerobic methane oxidation (n-damo), which couples the anaerobic oxidation of methane to denitrification, is a recently discovered process observed in Candidatus Methylomirabilis oxyfera. M. oxyfera is affiliated with the NC10 phylum, a phylum having no members in pure culture. Based on the isotopic labeling experiments, it is hypothesized that M. oxyfera has an unusual intra-aerobic pathway for the production of oxygen via the dismutation of nitric oxide into dinitrogen ga...

  18. Anaerobic biodegradation of PAHs in mangrove sediment with amendment of NaHCO3.

    Science.gov (United States)

    Li, Chun-Hua; Wong, Yuk-Shan; Wang, Hong-Yuan; Tam, Nora Fung-Yee

    2015-04-01

    Mangrove sediment is unique in chemical and biological properties. Many of them suffer polycyclic aromatic hydrocarbon (PAH) contamination. However, the study on PAH biological remediation for mangrove sediment is deficient. Enriched PAH-degrading microbial consortium and electron acceptor amendment are considered as two effective measures. Compared to other electron acceptors, the study on CO2, which is used by methanogens, is still seldom. This study investigated the effect of NaHCO3 amendment on the anaerobic biodegradation of four mixed PAHs, namely fluorene (Fl), phenanthrene (Phe), fluoranthene (Flua) and pyrene (Pyr), with or without enriched PAH-degrading microbial consortium in mangrove sediment slurry. The trends of various parameters, including PAH concentrations, microbial population size, electron-transport system activities, electron acceptor and anaerobic gas production were monitored. The results revealed that the inoculation of enriched PAH-degrading consortium had a significant effect with half lives shortened by 7-13 days for 3-ring PAHs and 11-24 days for 4-ring PAHs. While NaHCO3 amendment did not have a significant effect on the biodegradation of PAHs and other parameters, except that CO2 gas in the headspace of experimental flasks was increased. One of the possible reasons is that mangrove sediment contains high concentrations of other electron acceptors which are easier to be utilized by anaerobic bacteria, the other one is that the anaerobes in mangrove sediment can produce enough CO2 gas even without adding NaHCO3.

  19. Nitrate removal by organotrophic anaerobic ammonium oxidizing bacteria with C2/C3 fatty acid in upflow anaerobic sludge blanket reactors.

    Science.gov (United States)

    Liang, Yuhai; Li, Dong; Zhang, Xiaojing; Zeng, Huiping; Yang, Yin; Zhang, Jie

    2015-10-01

    In anaerobic ammonium oxidation (Anammox) process, a harsh ratio of nitrite to ammonia in influent was demanded, and the max nitrogen removal efficiency could only achieve to 89%, both of which limited the development of Anammox. The aim of this work was to study the nitrate removal by organotrophic anaerobic ammonium oxidizing bacteria (AAOB) with C2/C3 fatty acid in upflow anaerobic sludge blanket (UASB) reactors. In this study, organotrophic AAOB was successfully enriched by adding acetate and propionate with the total organic carbon to nitrogen (TOC/N) ratio of 0.1. In the condition of low substrate, the TN removal efficiency reached 90%, with the effluent TN of around 11.8 mg L(-1). After the addition of acetate and propionate, the predominant species in Anammox granular sludge transformed to Candidatus Jettenia that belonging to organotrophic AAOB from the Candidatus Kuenenia relating to general AAOB.

  20. Recalcitrant Carbonaceous Material: A Source of Electron Donors for Anaerobic Microbial Metabolisms in the Subsurface?

    Science.gov (United States)

    Nixon, S. L.; Montgomery, W.; Sephton, M. A.; Cockell, C. S.

    2014-12-01

    More than 90% of organic material on Earth resides in sedimentary rocks in the form of kerogens; fossilized organic matter formed through selective preservation of high molecular weight biopolymers under anoxic conditions. Despite its prevalence in the subsurface, the extent to which this material supports microbial metabolisms is unknown. Whilst aerobic microorganisms are known to derive energy from kerogens within shales, utilization in anaerobic microbial metabolisms that proliferate in the terrestrial subsurface, such as microbial iron reduction, has yet to be demonstrated. Data are presented from microbial growth experiments in which kerogens and shales were supplied as the sole electron donor source for microbial iron reduction by an enrichment culture. Four well-characterized kerogens samples (representative of Types I-IV, classified by starting material), and two shale samples, were assessed. Organic analysis was carried out to investigate major compound classes present in each starting material. Parallel experiments were conducted to test inhibition of microbial iron reduction in the presence of each material when the culture was supplied with a full redox couple. The results demonstrate that iron-reducing microorganisms in this culture were unable to use kerogens and shales as a source of electron donors for energy acquisition, despite the presence of compound classes known to support this metabolism. Furthermore, the presence of these materials was found to inhibit microbial iron reduction to varying degrees, with some samples leading to complete inhibition. These results suggest that recalcitrant carbonaceous material in the terrestrial subsurface is not available for microbial iron reduction and similar metabolisms, such as sulphate-reduction. Further research is needed to investigate the inhibition exerted by these materials, and to assess whether these findings apply to other microbial consortia. These results may have significant implications for

  1. Characterization of anaerobic chloroethene-dehalogenating activity in several subsurface sediments

    Energy Technology Data Exchange (ETDEWEB)

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

    1996-11-01

    Anaerobic microcosms of subsurface soils from four locations were used to investigate the separate effects of several electron donors on tetrachloroethylene (PCE) dechlorination activity. The substrates tested were methanol, formate, lactate, acetate, and sucrose. Various levels of sulfate-reducing, acetogenic, fermentative, and methanogenic activity were observed in all sediments. PCE dechlorination was detected in all microcosms, but the amount of dehalogenation varied by several orders of magnitude. Trichloroethylene was the primary dehalogenation product; however, small amounts of cis-1,2-dichloroethylene, 1,1-dichloroethylene, and vinyl chloride were also detected in several microcosms. Lactate-amended microcosms showed large amounts of dehalogenation. in three of the four sediments. One of the two sediments which showed positive activity with lactate also had large amounts of delialogenation with methanol. Sucrose, formate, and acetate also stimulated large amounts of delialogenation in one sediment that showed activity with lactate. These results suggest that lactate may be an appropriate substrate for screening sediments for PCE or TCE delialogenation activity, but that the microbial response is not sufficient for complete in situ bioremediation. A detailed study of the Victoria activity revealed that delialogenation rates were more similar to the Cornell culture than to rates measured for methanogens, or a methanol-enriched sediment culture. This may suggest that these sediments contain a highly efficient delialogenation activity similar to the Cornell culture. This assertion is supported further by the fact that an average of 3% of added reducing equivalents could be diverted to dehalogenation in tests which were conducted using PCE-saturated hexadecane as a constant source of PCE during incubation. Further evidence is needed to confirm this premise. The application of these results to in situ bioremediation of highly contaminated areas are discussed.

  2. Degradation of TCE using sequential anaerobic biofilm and aerobic immobilized bed reactor

    Science.gov (United States)

    Chapatwala, Kirit D.; Babu, G. R. V.; Baresi, Larry; Trunzo, Richard M.

    1995-01-01

    Bacteria capable of degrading trichloroethylene (TCE) were isolated from contaminated wastewaters and soil sites. The aerobic cultures were identified as Pseudomonas aeruginosa (four species) and Pseudomonas fluorescens. The optimal conditions for the growth of aerobic cultures were determined. The minimal inhibitory concentration values of TCE for Pseudomonas sps. were also determined. The aerobic cells were immobilized in calcium alginate in the form of beads. Degradation of TCE by the anaerobic and dichloroethylene (DCE) by aerobic cultures was studied using dual reactors - anaerobic biofilm and aerobic immobilized bed reactor. The minimal mineral salt (MMS) medium saturated with TCE was pumped at the rate of 1 ml per hour into the anaerobic reactor. The MMS medium saturated with DCE and supplemented with xylenes and toluene (3 ppm each) was pumped at the rate of 1 ml per hour into the fluidized air-uplift-type reactor containing the immobilized aerobic cells. The concentrations of TCE and DCE and the metabolites formed during their degradation by the anaerobic and aerobic cultures were monitored by GC. The preliminary study suggests that the anaerobic and aerobic cultures of our isolates can degrade TCE and DCE.

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

    OpenAIRE

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

    1983-01-01

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

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

    Science.gov (United States)

    Gövert, D.; Conrad, R.

    2009-04-01

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

  5. Consideration and development of anaerobic fermentation equipment on solid organic rejectamenta

    Institute of Scientific and Technical Information of China (English)

    2007-01-01

    This article, based on investigation and analysis, existed anaerobic fermentation equipment, with a view to the production of high-latitude area, pointed out the thought of exploiting efficient anaerobic fermentation equipment, including: the settling independent equipment of acidogenic phase and methanogenic phase; the applying conbination of AF and UASB in methanogenic phase; adopting efficient sludge inverse flowing equipment and the technique of flora enrichment, and efficient method of saving energy and thermal retardation; adopting autocontrol which could make the equipment run efficiently and stably.

  6. Uranium Conversion & Enrichment

    Energy Technology Data Exchange (ETDEWEB)

    Karpius, Peter Joseph [Los Alamos National Lab. (LANL), Los Alamos, NM (United States)

    2017-02-06

    The isotopes of uranium that are found in nature, and hence in ‘fresh’ Yellowcake’, are not in relative proportions that are suitable for power or weapons applications. The goal of conversion then is to transform the U3O8 yellowcake into UF6. Conversion and enrichment of uranium is usually required to obtain material with enough 235U to be usable as fuel in a reactor or weapon. The cost, size, and complexity of practical conversion and enrichment facilities aid in nonproliferation by design.

  7. Potential for anaerobic conversion of xenobiotics

    DEFF Research Database (Denmark)

    Mogensen, Anders Skibsted; Dolfing, J.; Haagensen, Frank;

    2003-01-01

    This review covers the latest research on the anaerobic biodegradation of aromatic xenobiotic compounds, with emphasis on surfactants, polycyclic aromatic hydrocarbons, phthalate esters, polychlorinated biphenyls, halogenated phenols, and pesticides. The versatility of anaerobic reactor systems...

  8. Porphyromonas gingivalis as a Model Organism for Assessing Interaction of Anaerobic Bacteria with Host Cells.

    Science.gov (United States)

    Wunsch, Christopher M; Lewis, Janina P

    2015-12-17

    Anaerobic bacteria far outnumber aerobes in many human niches such as the gut, mouth, and vagina. Furthermore, anaerobic infections are common and frequently of indigenous origin. The ability of some anaerobic pathogens to invade human cells gives them adaptive measures to escape innate immunity as well as to modulate host cell behavior. However, ensuring that the anaerobic bacteria are live during experimental investigation of the events may pose challenges. Porphyromonas gingivalis, a Gram-negative anaerobe, is capable of invading a variety of eukaryotic non-phagocytic cells. This article outlines how to successfully culture and assess the ability of P. gingivalis to invade human umbilical vein endothelial cells (HUVECs). Two protocols were developed: one to measure bacteria that can successfully invade and survive within the host, and the other to visualize bacteria interacting with host cells. These techniques necessitate the use of an anaerobic chamber to supply P. gingivalis with an anaerobic environment for optimal growth. The first protocol is based on the antibiotic protection assay, which is largely used to study the invasion of host cells by bacteria. However, the antibiotic protection assay is limited; only intracellular bacteria that are culturable following antibiotic treatment and host cell lysis are measured. To assess all bacteria interacting with host cells, both live and dead, we developed a protocol that uses fluorescent microscopy to examine host-pathogen interaction. Bacteria are fluorescently labeled with 2',7'-Bis-(2-carboxyethyl)-5-(and-6)-carboxyfluorescein acetoxymethyl ester (BCECF-AM) and used to infect eukaryotic cells under anaerobic conditions. Following fixing with paraformaldehyde and permeabilization with 0.2% Triton X-100, host cells are labeled with TRITC phalloidin and DAPI to label the cell cytoskeleton and nucleus, respectively. Multiple images taken at different focal points (Z-stack) are obtained for temporal

  9. Arsenic, Anaerobes, and Astrobiology

    Science.gov (United States)

    Stolz, J. F.; Oremland, R. S.; Switzer Blum, J.; Hoeft, S. E.; Baesman, S. M.; Bennett, S.; Miller, L. G.; Kulp, T. R.; Saltikov, C.

    2013-12-01

    Arsenic is an element best known for its highly poisonous nature, so it is not something one would associate with being a well-spring for life. Yet discoveries made over the past two decades have delineated that not only are some microbes resistant to arsenic, but that this element's primary redox states can be exploited to conserve energy and support prokaryotic growth ('arsenotrophy') in the absence of oxygen. Hence, arsenite [As(III)] can serve as an electron donor for chemo- or photo-autotrophy while arsenate [As(V)] will serve as an electron acceptor for chemo-heterotrophs and chemo-autotrophs. The phylogenetic diversity of these microbes is broad, encompassing many individual species from diverse taxonomic groups in the Domain Bacteria, with fewer representatives in the Domain Archaea. Speculation with regard to the evolutionary origins of the key functional genes in anaerobic arsenic transformations (arrA and arxA) and aerobic oxidation (aioB) has led to a disputation as to which gene and function is the most ancient and whether arsenic metabolism extended back into the Archaean. Regardless of its origin, robust arsenic metabolism has been documented in extreme environments that are rich in their arsenic content, such as hot springs and especially hypersaline soda lakes associated with volcanic regions. Searles Lake, CA is an extreme, salt-saturated end member where vigorous arsenic metabolism occurs, but there is no detectable sulfate-reduction or methanogenesis. The latter processes are too weak bio-energetically to survive as compared with arsenotrophy, and are also highly sensitive to the abundance of borate ions present in these locales. These observations have implications with respect to the search for microbial life elsewhere in the Solar System where volcanic-like processes have been operative. Hence, because of the likelihood of encountering dense brines in the regolith of Mars (formed by evapo-concentration) or beneath the ice layers of Europa

  10. A Cultural Classroom Library

    Science.gov (United States)

    Lawrence, Maria

    2007-01-01

    Native American and other cultural stories provide students with a broader perspective on the world. In addition, cultural stories connect science content and knowledge about the world to cultural interpretations and people's life ways. By implementing the ideas suggested in this article, you can select books that both enrich your science library…

  11. Anaerobic methane oxidation in a landfill-leachate plume.

    Science.gov (United States)

    Grossman, Ethan L; Cifuentes, Luis A; Cozzarelli, Isabelle M

    2002-06-01

    The alluvial aquifer adjacent to Norman Landfill, OK, provides an excellent natural laboratory for the study of anaerobic processes impacting landfill-leachate contaminated aquifers. We collected groundwaters from a transect of seven multilevel wells ranging in depth from 1.3 to 11 m that were oriented parallel to the flow path. The center of the leachate plume was characterized by (1) high alkalinity and elevated concentrations of total dissolved organic carbon, reduced iron, and methane, and (2) negligible oxygen, nitrate, and sulfate concentrations. Methane concentrations and stable carbon isotope (delta13C) values suggest anaerobic methane oxidation was occurring within the plume and at its margins. Methane delta13C values increased from about -54 per thousand near the source to > -10 per thousand downgradient and at the plume margins. The isotopic fractionation associated with this methane oxidation was -13.6+/-1.0 per thousand. Methane 13C enrichment indicated that 80-90% of the original landfill methane was oxidized over the 210-m transect. First-order rate constants ranged from 0.06 to 0.23 per year, and oxidation rates ranged from 18 to 230 microM/y. Overall, hydrochemical data suggest that a sulfate reducer-methanogen consortium may mediate this methane oxidation. These results demonstrate that natural attenuation through anaerobic methane oxidation can be an important sink for landfill methane in aquifer systems.

  12. Bioconversion of selenate in methanogenic anaerobic granular sludge.

    Science.gov (United States)

    Astratinei, Violeta; van Hullebusch, Eric; Lens, Piet

    2006-01-01

    The capacity of anaerobic granular sludge to remove selenate from contaminated wastewater was investigated. The potential of different types of granular sludge to remove selenate from the liquid phase was compared to that of suspended sludge and contaminated soil and sediment samples. The selenate removal rates ranged from 400 to 1500 microg g VSS(-1) h(-1), depending on the source of biomass, electron donor, and the initial selenate concentration. The granular structure protects the microorganisms when exposed to high selenate concentrations (0.1 to 1 mM). Anaerobic granular sludge "Eerbeek," originating from a UASB reactor treating paper mill wastewater, removed about 90, 50, and 36% of 0.1, 0.5, and 1 mM of Se, respectively, from the liquid phase when incubated with 20 mM lactate at 30 degrees C and pH 7.5. Selenite, elemental Se (Se(o)), and metal selenide precipitates were the conversion products. Enrichments from the anaerobic granular sludge "Eerbeek" were able to convert 90% of the 10-mM selenate to Se(o) at a rate of 1505 microg Se(VI) g cells(-1) h(-1), a specific growth rate of 0.0125 g cells h(-1), and a yield of 0.083 g cells mg Se(-1). Both microbial metabolic processes (e.g dissimilatory reduction) as well as microbially mediated physicochemical mechanisms (adsorption and precipitation) contribute to the removal of selenate from the Se-containing medium.

  13. Enriching the Catalog

    Science.gov (United States)

    Tennant, Roy

    2004-01-01

    After decades of costly and time-consuming effort, nearly all libraries have completed the retrospective conversion of their card catalogs to electronic form. However, bibliographic systems still are really not much more than card catalogs on wheels. Enriched content that Amazon.com takes for granted--such as digitized tables of contents, cover…

  14. 21 CFR 866.2120 - Anaerobic chamber.

    Science.gov (United States)

    2010-04-01

    ... and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES IMMUNOLOGY AND MICROBIOLOGY DEVICES Microbiology Devices § 866.2120 Anaerobic chamber. (a) Identification. An anaerobic chamber is a device intended for medical purposes to maintain an anaerobic...

  15. The phenomenon of granulation of anaerobic sludge.

    NARCIS (Netherlands)

    Hulshoff Pol, L.W.

    1989-01-01

    Successful high-rate anaerobic wastewater treatment can only be accomplished when the slowgrowing anaerobic biomass is efficiently held back in the anaerobic treatment system. This biomass retention can be achieved in various ways including immobilization of the organisms on fixed materials and immo

  16. Involvement of protozoa in anaerobic wastewater treatment process.

    Science.gov (United States)

    Priya, M; Haridas, Ajit; Manilal, V B

    2007-12-01

    It is only very rarely recognised in literature that anaerobic reactors may contain protozoa in addition to various bacterial and archeal groups. The role of protozoa in anaerobic degradation was studied in anaerobic continuous stirred tank reactors (CSTR) and batch tests. Anaerobic protozoa, especially the ciliated protozoa, have direct influence on the performance of CSTR at all organic loading rates (1-2g CODl(-1)d(-1)) and retention times (5-10 days). The studies revealed that chemical oxygen demand (COD) removal is strongly correlated to ciliate density in CSTR fed with oleate (suspended COD) and acetate (soluble COD). There was no significant difference in COD removal between reactors fed suspended COD and those fed soluble COD. However, the diversity and number of ciliates is greater in CSTR fed with particulate feed. The mixed liquor suspended solids (MLSS) representing biomass was significantly lower (16-34%) in CSTR with protozoa. In batch tests, increased COD removal and methane production was observed in sludge having ciliates as compared with sludge without protozoa. Methane production increased linearly with number of ciliates (R(2)=0.96) in batch tests with protozoa. Direct utilization of COD by flagellates and ciliates was observed in bacteria-suppressed cultures. The technological importance of these results is that reactors with protozoa-rich sludge can enhance the rate of mineralization of complex wastewater, especially wastewater containing particulate COD.

  17. Bacterial community succession during the enrichment of chemolithoautotrophic arsenite oxidizing bacteria at high arsenic concentrations

    Institute of Scientific and Technical Information of China (English)

    Nguyen Ai Le; Akiko Sato; Daisuke Inoue; Kazunari Sei; Satoshi Soda; Michihiko Ike

    2012-01-01

    To generate cost-effective technologies for the removal of arsenic from water,we developed an enrichment culture of chemolithoautotrophic arsenite oxidizing bacteria (CAOs) that could effectively oxidize widely ranging concentrations of As(Ⅲ) to As(Ⅴ).In addition,we attempted to elucidate the enrichment process and characterize the microbial composition of the enrichment culture.A CAOs enrichment culture capable of stably oxidizing As(Ⅲ) to As(Ⅴ) was successfully constructed through repeated batch cultivation for more than 700 days,during which time the initial As(Ⅲ) concentrations were increased in a stepwise manner from l to 10-12 mmol/L.As(Ⅲ) oxidation activity of the enrichment culture gradually improved,and 10-12 mmol/L As(Ⅲ) was almost completely oxidized within four days.Terminal restriction fragment length polymorphism analysis showed that the dominant bacteria in the enrichment culture varied drastically during the enrichment process depending on the As(Ⅲ) concentration.Isolation and characterization of bacteria in the enrichment culture revealed that the presence of multiple CAOs with various As(Ⅲ) oxidation abilities enabled the culture to adapt to a wide range of As(Ⅲ) concentrations.The CAOs enrichment culture constructed here may he useful for pretreatment of water from which arsenic is being removed.

  18. Anaerobic nitrite-dependent methane-oxidizing bacteria - novel participants in methane cycling of drained peatlands ecosystems

    Science.gov (United States)

    Kravchenko, Irina; Sukhacheva, Marina; Menko, Ekaterina; Sirin, Andrey

    2014-05-01

    amplification of 16S rRNA (Ettwig et al. 2009) and pmoA (Luesken et al. 2011) genes followed by construction of clone libraries. Phylogenetic analysis revealed only one n-damo bacterium distantly related to uncultured anaerobic methanotrophs found in situ. It may represent a new cluster of NC10 bacteria with an identity of less than 96 and 86% to the 16S rRNA and pmoA genes of "Ca. Methylomirabilis oxyfera," respectively. An enrichment of nitrite-reducing methanotrophic NC10 bacteria was successfully obtained from this sample in a static anaerobic culture with methane and nitrite at an in situ pH of 6.3. The bacterial abundance in enrichment was estimated using quantitative PCR and FISH (DBACT-0193-a-A probe) analysis and was found to increase up to 10 times for 120 days. The results of this study expand our knowledge of the diversity and distribution of NC10 bacteria in the environment and their potential contribution to nitrogen and methane cycles in northern peatland ecosystems. We think that AOM may be more active in anthropogenic disturbed peatlands with greater supply of elements that could potentially serve as electron acceptors. In spite of generally low concentration, seasonal increases in nitrate content in drained peatlands may work as an important control of CH4 fluxes. The study was partially supported by RFBR research project # 12-05-01029_a.

  19. Perspectives of Anaerobic Soil Disinfestation

    NARCIS (Netherlands)

    Lamers, J.G.; Runia, W.T.; Molendijk, L.P.G.; Bleeker, P.O.

    2010-01-01

    Biological soil disinfestation is an environmentally friendly method to disinfest soil. From now on we refer to it as anaerobic soil disinfestation (ASD). With ASD a green manure crop (40 t/ha) is homogeneously incorporated into the topsoil (0-30 cm) after which the field is lightly compacted and ir

  20. Anaerobic granular sludge and biofilm reactors

    DEFF Research Database (Denmark)

    Skiadas, Ioannis V.; Gavala, Hariklia N.; Schmidt, Jens Ejbye

    2003-01-01

    by the immobilization of the biomass, which forms static biofilms, particle-supported biofilms, or granules depending on the reactor's operational conditions. The advantages of the high-rate anaerobic digestion over the conventional aerobic wastewater treatment methods has created a clear trend for the change......-rate anaerobic treatment systems based on anaerobic granular sludge and biofilm are described in this chapter. Emphasis is given to a) the Up-flow Anaerobic Sludge Blanket (UASB) systems, b) the main characteristics of the anaerobic granular sludge, and c) the factors that control the granulation process...

  1. Polyphasic characterization of a PCP-to-phenol dechlorinating microbial community enriched from paddy soil

    Energy Technology Data Exchange (ETDEWEB)

    Yoshida, Naoko [EcoTopia Science Institute, Nagoya University Nagoya 464-8603 (Japan)]. E-mail: ysd75@esi.nagoya-u.ac.jp; Yoshida, Yukina [Graduate School of Engineering, Nagoya University, Nagoya 464-8603 (Japan); Handa, Yuko [Graduate School of Engineering, Nagoya University, Nagoya 464-8603 (Japan); Kim, Hyo-Keun [Korea Ginseng and Tobacco Research Institute, Taejon 305-345 (Korea, Republic of); Ichihara, Shigeyuki [Faculty of Agriculture, Meijo University, Nagoya 468-8502 (Japan); Katayama, Arata [EcoTopia Science Institute, Nagoya University Nagoya 464-8603 (Japan); Graduate School of Engineering, Nagoya University, Nagoya 464-8603 (Japan)

    2007-08-01

    Dechlorination of PCP has been observed previously under anaerobic condition in paddy soil. However, there is poor information about the dechlorination pathway of PCP and the microbial community associated with the PCP dechlorination in paddy soil. In this study, an anaerobic microbial community dechlorinating PCP was enriched by serial transfers from a paddy soil using a medium containing PCP, lactate and the steam-sterilized paddy soil. The enriched microbial community dechlorinated PCP completely to phenol under the anaerobic condition by a dechlorinating pathway as follows; PCP {sup {yields}} 2,3,4,5-tetrachlorophenol {sup {yields}} 3,4,5-trichlorophenol {sup {yields}} 3,5-dichlorophenol {sup {yields}} 3-chlorophenol {sup {yields}} phenol. Intermediate products such as 3-chlorophenol were not accumulated, which were immediately dechlorinated to phenol. The enriched microbial community was characterized physiologically by testing the effects of electron donors and electron acceptors on the dechlorinating activity. The dechlorinating activity was promoted with lactate, pyruvate, and hydrogen as electron donors but not with acetate. Electron acceptors, nitrate and sulphate, inhibited the dechlorinating activity competitively but not iron (III). The microbial group associated with the anaerobic dechlorination was characterized by the effect of specific inhibitors on the PCP dechlorination. Effects of specific metabolic inhibitors and antibiotics indicated the involvement of Gram-positive spore-forming bacteria with the PCP dechlorinating activity, which was represented as bacteria of phylum Firmicutes. The structure of the microbial community was characterized by fluorescence in situ hybridization, quinone profiling, and PCR-DGGE (denaturing gel gradient electrophoresis). The combined results indicated the predominance of Clostridium species of phylum Firmicutes in the microbial community. Desulfitobacterium spp. known as anaerobic Gram-positive spore

  2. Investigation of the effect of culture type on biological hydrogen production from sugar industry wastes.

    Science.gov (United States)

    Ozkan, Leyla; Erguder, Tuba H; Demirer, Goksel N

    2010-05-01

    The bio-hydrogen generation potential of sugar industry wastes was investigated. In the first part of the study, acidogenic anaerobic culture was enriched from the mixed anaerobic culture (MAC) through acidification of glucose. In the second part of the study, glucose acclimated acidogenic seed was used, along with the indigenous microorganisms, MAC, 2-bromoethanesulfonate treated MAC and heat treated MAC. Two different COD levels (4.5 and 30g/L COD) were investigated for each culture type. Reactors with initial COD concentration of 4.5g/L had higher H(2) yields (20.3-87.7mL H(2)/g COD) than the reactors with initial COD concentration of 30g/L (0.9-16.6mL H(2)/g COD). The 2-bromoethanesulfonate and heat treatment of MAC inhibited the methanogenic activity, but did not increase the H(2) production yield. The maximum H(2) production (87.7mL H(2)/g COD) and minimum methanogenic activity were observed in the unseeded reactor with 4.5g/L of initial COD.

  3. Effect of anaerobic bovine colostrum fermentation on bacteria growth inhibition

    Directory of Open Access Journals (Sweden)

    Mara Helena Saalfeld

    Full Text Available ABSTRACT: Efficient handling programs that provide high quality colostrum in adequate amounts to dairy farm calves are needed to assure their health and survival. Replacers (or milk substitutes often become necessary when colostrum presents inadequate quality, or in order to break the cycle of infectious disease transmission. In this study we aimed to assess the effect of anaerobic fermentation processing (colostrum silage on bacterial that represent interest to animal health. Colostrum samples were inoculated with cultures of Brucella abortus , Escherichia coli , Leptospira interrogans serovar Copenhageni , Mycobacterium bovis , Salmonella Enteritidis , Salmonella Typhimurium , Staphylococcus aureus , and Bacillus cereus and then subjected to anaerobic fermentation. On the first day, and every seven days until 30th days after fermentation, the samples were cultured and colony forming units counted. At seven days of fermentation, B. abortus , L. interrogans , and M. bovis were not detected. At 14th days of fermentation, E. coli , S. aureus , S. Enteritidis and S. Typhimurium were no longer detected. However, we were able to detect both lactic acid bacteria and B. cereus until 30th days of fermentation. From this study we suggested that anaerobic fermentation processing can inhibit important bacteria that cause economical losses for the cattle industry. The observations suggested that colostrum silage is a promising form to conserve bovine colostrum.

  4. Anaerobic degradation of benzoate by sulfate-reducing bacteria

    Energy Technology Data Exchange (ETDEWEB)

    Silva, S.P.; Adorno, M.A.T.; Moraes, E.M.; Varesche, M.B.A. [Sao Paulo Univ., Sao Carlos, SP (Brazil). Biological Processes Laboratory

    2004-07-01

    Anaerobic processes are an efficient way to degrade aromatic compounds in industrial wastewater, such as phenol, cresol and benzoate. This study characterized the bacteria that degrades benzoate, an anaerobic degradation intermediate of several complex aromatic compounds. In particular, the study assessed the capacity to use benzoate with sulfate reducing bacteria in mesophilic conditions. Biofilm from polyurethane foam matrices of a fixed bed reactor was used as the cellular inoculum to treat industrial wastewater containing organic peroxide. Dilution techniques were used to purify the material and obtain cultures of cocci. The benzoate consumption capacity in sulfidogenic conditions was observed when the purified inoculum was applied to batch reactors with different benzoate/sulfate relations. Results indicate that purification was positive to bacteria that can degrade aromatic compounds. Desulfococcus multivorans bacteria was identified following the physiologic and kinetic experiments. The 0.6 benzoate/sulfate relation was considered ideal for complete consumption of carbon and total use of sulfur. 10 refs., 3 figs.

  5. Binary Interactions of Antagonistic Bacteria with Candida albicans Under Aerobic and Anaerobic Conditions.

    Science.gov (United States)

    Benadé, Eliska; Stone, Wendy; Mouton, Marnel; Postma, Ferdinand; Wilsenach, Jac; Botha, Alfred

    2016-04-01

    We used both aerobic and anaerobic liquid co-cultures, prepared with Luria Bertani broth, to study the effect of bacteria on the survival of Candida albicans in the external environment, away from an animal host. The bacteria were represented by Aeromonas hydrophila, Bacillus cereus, Bacillus subtilis, Clostridium, Enterobacter, Klebsiella pneumoniae, Kluyvera ascorbata and Serratia marcescens. Under aerobic conditions, the yeast's growth was inhibited in the presence of bacterial growth; however, under anaerobic conditions, yeast and bacterial growth in co-cultures was similar to that observed for pure cultures. Subsequent assays revealed that the majority of bacterial strains aerobically produced extracellular hydrolytic enzymes capable of yeast cell wall hydrolysis, including chitinases and mannan-degrading enzymes. In contrast, except for the A. hydrophila strain, these enzymes were not detected in anaerobic bacterial cultures, nor was the antimicrobial compound prodigiosin found in anaerobic cultures of S. marcescens. When we suspended C. albicans cells in crude extracellular enzyme preparations from K. pneumoniae and S. marcescens, we detected no negative effect on yeast viability. However, we found that these preparations enhance the toxicity of prodigiosin towards the yeast, especially in combination with mannan-degrading enzymes. Analyses of the chitin and mannan content of yeast cell walls revealed that less chitin was produced under anaerobic than aerobic conditions; however, the levels of mannan, known for its low permeability, remained the same. The latter phenomenon, as well as reduced production of the bacterial enzymes and prodigiosin, may contribute to anaerobic growth and survival of C. albicans in the presence of bacteria.

  6. Occurrence of fastidious Campylobacter spp. in fresh meat and poultry using an adapted cultural protocol.

    Science.gov (United States)

    Lynch, Orla A; Cagney, Claire; McDowell, David A; Duffy, Geraldine

    2011-11-01

    This study used an adapted cultural protocol for the recovery of fastidious species of Campylobacter, to gain a more accurate understanding of the diversity of Campylobacter populations in fresh meats. Chicken (n=185), pork (n=179) and beef (n=186) were collected from supermarkets and butchers throughout the Republic of Ireland. Samples were enriched in Campylobacter enrichment broth for 24h under an atmosphere of 2.5% O(2), 7% H(2), 10% CO(2), and 80.5% N(2). The enriched samples were then filtered onto non-selective Anaerobe Basal Agar supplemented with lysed horse blood using mixed ester filter membranes. Isolates were identified by both genus and species-specific PCR assays and biochemical testing. The incidence of campylobacters on beef (36%) was significantly higher than on pork (22%) or chicken (16%), and far exceeds previously reported prevalence levels. The method was successful in recovering 7 species of Campylobacter, including the fastidious spp. C. concisus and C. mucosalis, from chicken meat, and 10 species, including C. concisus, C. curvus, C. mucosalis, C. sputorum, and C. upsaliensis, from minced beef. The isolation of C. concisus and C. upsaliensis from meat in this study is of particular significance, due to their emerging clinical relevance. The results of this study confirm that the diversity of Campylobacter species on fresh meats is greater than previously reported and highlights the bias of cultural methods towards the recovery of C. jejuni.

  7. The Effect of Anaerobic and Aerobic Fish Sludge Supernatant on Hydroponic Lettuce

    Directory of Open Access Journals (Sweden)

    Simon Goddek

    2016-06-01

    Full Text Available The mobilization of nutrients from fish sludge (i.e., feces and uneaten feed plays a key role in optimizing the resource utilization and thus in improving the sustainability of aquaponic systems. While several studies have documented the aerobic and anaerobic digestion performance of aquaculture sludge, the impact of the digestate on plant growth has yet to be understood. The present study examines the impact of either an aerobic or an anaerobic digestion effluent on lettuce plant growth, by enriching a mixture of aquaculture and tap water with supernatants from both aerobic and anaerobic batch reactors. The lettuce plants grown in the hydroponic system supplied with supernatant from an anaerobic reactor had significantly better performance with respect to weight gain than both, those in the system where supernatant from the aerobic reactor was added, as well as the control system. It can be hypothesized that this effect was caused by the presence of NH4+ as well as dissolved organic matter, plant growth promoting rhizobacteria and fungi, and humic acid, which are predominantly present in anaerobic effluents. This study should therefore be of value to researchers and practitioners wishing to further develop sludge remineralization in aquaponic systems.

  8. Molecular ecology of anaerobic reactor systems

    DEFF Research Database (Denmark)

    Hofman-Bang, H. Jacob Peider; Zheng, D.; Westermann, Peter;

    2003-01-01

    Anaerobic reactor systems are essential for the treatment of solid and liquid wastes and constitute a core facility in many waste treatment plants. Although much is known about the basic metabolism in different types of anaerobic reactors, little is known about the microbes responsible...... to the abundance of each microbe in anaerobic reactor systems by rRNA probing. This chapter focuses on various molecular techniques employed and problems encountered when elucidating the microbial ecology of anaerobic reactor systems. Methods such as quantitative dot blot/fluorescence in-situ probing using various...

  9. Evaluation of a prereduced anaerobically sterilized medium (PRAS II) system for identification anaerobic microorganisms.

    Science.gov (United States)

    Beaucage, C M; Onderdonk, A B

    1982-09-01

    A prereduced, anaerobically sterilized system of tubed media (PRAS II; Scott Laboratories, Fiskeville, R.I.) was evaluated for accuracy in the identification of anerobic microorganisms. PRAS II was found to be a rapid and accurate identification system for obligate anaerobes which does not require the use of gas cannula inoculation or incubation in a special anaerobic environment.

  10. Evaluation of a prereduced anaerobically sterilized medium (PRAS II) system for identification anaerobic microorganisms.

    OpenAIRE

    Beaucage, C M; Onderdonk, A B

    1982-01-01

    A prereduced, anaerobically sterilized system of tubed media (PRAS II; Scott Laboratories, Fiskeville, R.I.) was evaluated for accuracy in the identification of anerobic microorganisms. PRAS II was found to be a rapid and accurate identification system for obligate anaerobes which does not require the use of gas cannula inoculation or incubation in a special anaerobic environment.

  11. Applications of aerobic and anaerobic bacteria in the fields of biological degradation of contaminants and biological wastewater treatment

    OpenAIRE

    Jovcic, Alexander

    2003-01-01

    In the work here presented four distinctly different problems were investigated. The first problem was an investigation into the degradation of Dichloroethylene (DCE) and 1,1-bis (p-Chlorophenyl)-2-dichloroethylene (DDE) utilising pure bacterial cultures. The second investigation dealt with the degradation of DDE and polychlorinated Biphenyl’s (PCB’s) utilising anaerobic sediments and soils from New Zealand. The third investigation worked on the Granulation of anaerobic River-sediments in Upf...

  12. 新型OxyPlateTM厌氧系统隔离眼部厌氧菌的评估%Evaluation of the new OxyPlateTM Anaerobic System for the isolation of ocular anaerobic bacteria

    Institute of Scientific and Technical Information of China (English)

    Emily K. Deschler; Paul P. Thompson; Regis Paul Kowalski

    2013-01-01

    目的:厌氧细菌可引起眼部感染,我们测试OxyPlateTM厌氧系统(OxyPlateTM Anaerobic System,OXY)隔离可引起眼部疾病的厌氧细菌.方法:OXY不需要直接的厌氧条件(比如厌氧袋,罐),将其与常规的厌氧袋培养基相比.琼脂培养基上眼部厌氧细菌菌株在好氧和厌氧条件下(厌氧袋)行标准的菌落计数:(1)OXY(好氧);(2)5%羊血(sheep blood,SB);(3)巧克力琼脂;(4)Schaedler琼脂.测试的眼部体外培养细菌来自眼内炎,泪囊炎,包括10个丙酸杆菌和3个放线菌种类.在每个培养条件下,每个细菌菌落计数隔离,排名从大到小,并在非参数比较下确定最佳的培养条件.结果:所有的厌氧条件对于厌氧菌株呈阳性反应.厌氧菌在有氧条件下的SB和Schaedler的琼脂中无法增长.痤疮丙酸杆菌在巧克力琼脂中生长稀疏.作为一种厌氧系统,在厌氧袋SB分离比OXY(P=0.0028)和巧克力琼脂(P=0.0028)分离出更多的菌落数.结论:虽然OXY经测试并没比其他的厌氧系统更高效,它似乎是一个合理隔离厌氧细菌的替代方法.其琼脂培养基在一个专门设计的盘并不需要厌氧袋使得OXY优于其他厌氧系统.%AIM: Anaerobic bacteria can cause ocular infections. We tested the OxyPlateTM Anaerobic System (OXY) to isolate pertinent anaerobic bacteria that can cause ocular disease.METHODS: OXY, which does not require direct anaerobic conditions (i.e. bags, jars), was compared to conventional isolation of incubating culture media in anaerobic bags. Standard colonies counts were performed on anaerobic ocular bacterial isolates under aerobic and anaerobic conditions (anaerobic bags) using agar media: 1) OXY (aerobic only), 2) 5% sheep blood (SB), 3) Chocolate, and 4) Schaedler. The bacteria tested were de-identified ocular isolates cultured from endophthalmitis and dacryocystitis that include 10 Propionibacterium acnes and 3 Actinomyces species. The colony counts for each bacteria isolate, on each

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

    Science.gov (United States)

    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).

  14. EVALUATION OF THE TEA TREE OIL ACTIVITY TO ANAEROBIC BACTERIA--IN VITRO STUDY.

    Science.gov (United States)

    Ziółkowska-Klinkosz, Marta; Kedzia, Anna; Meissner, Hhenry O; Kedzia, Andrzej W

    2016-01-01

    The study of the sensitivity to tea tree oil (Australian Company TTD International Pty. Ltd. Sydney) was carried out on 193 strains of anaerobic bacteria isolated from patients with various infections within the oral cavity and respiratory tracts. The susceptibility (MIC) of anaerobes was determined by means of plate dilution technique in Brucella agar supplemented with 5% defibrinated sheep blood, menadione and hemin. Inoculum contained 10(5) CFU per spot was cultured with Steers replicator upon the surface of agar with various tea tree oil concentrations or without oil (anaerobes growth control). Incubation the plates was performed in anaerobic jars under anaerobic conditions at 37 degrees C for 48 h. MIC was defined as the lowest concentrations of the essential oil completely inhibiting growth of anaerobic bacteria. Test results indicate, that among Gram-negative bacteria the most sensitive to essential oil were strains of Veillonella and Porphyromonas species. Essential oil in low concentrations (MIC in the range of = 0.12 - 0.5 mg/mL) inhibited growth of accordingly 80% and 68% strains. The least sensitive were strains of the genus Tannerella, Parabacteroides and Dialister (MIC 1.0 - 2.0 mg/mL). In the case of Gram-positive anaerobic bacteria the tea tree oil was the most active to strains of cocci of the genus Anaerococcus and Ruminococcus (MIC in range = 0.12 - 0.5 mg/mL) or strains of rods of the genus Eubacterium and Eggerthella (MIC = 0.25 mg/mL). Among Gram-positive rods the least sensitive were the strains of the genus Bifidobacterium ( MIC = 2.0 mg/mL). The tea tree oil was more active to Gram-positive than to Gram-negative anaerobic bacteria.

  15. NADH-linked aldose reductase: the key to anaerobic alcoholic fermentation of xylose by yeasts

    NARCIS (Netherlands)

    Bruinenberg, P.M.; De Bot, P.H.M.; Van Dijken, J.P.; Scheffers, W.A.

    1984-01-01

    The kinetics and enzymology of o-xylose utilization were studied in aerobic and anaerobic batch cultures of the facultatively fermentative yeasts Candida utilis, Pachysolen tannophilus, and Pichia stipitis. These yeasts did not produce ethanol under aerobic conditions. When shifted to anaerobiosis c

  16. ANAEROBIC DDT BIOTRANSFORMATION: ENHANCEMENT BY APPLICATION OF SURFACTANTS AND LOW OXIDATION REDUCTION POTENTIAL

    Science.gov (United States)

    Enhancement of anaerobic DDT (1,1,1-trichloro-2,2-bis(p-chlorophenyl) ethane) biotransformation by mixed cultures was studied with application of surfactants and oxidation reduction potential reducing agents. Without amendments, DDT transformation resulted mainly in the pr...

  17. Chain elongation in anaerobic reactor microbiomes to recover resources from waste

    NARCIS (Netherlands)

    Spirito, C.M.; Richter, H.; Rabaey, K.; Stams, A.J.M.; Angenent, L.T.

    2014-01-01

    Different microbial pathways can elongate the carbon chains of molecules in open cultures of microbial populations (i.e. reactor microbiomes) under anaerobic conditions. Here, we discuss three such pathways: 1. homoacetogenesis to combine two carbon dioxide molecules into acetate; 2. succinate forma

  18. Thermal breeder fuel enrichment zoning

    Science.gov (United States)

    Capossela, Harry J.; Dwyer, Joseph R.; Luce, Robert G.; McCoy, Daniel F.; Merriman, Floyd C.

    1992-01-01

    A method and apparatus for improving the performance of a thermal breeder reactor having regions of higher than average moderator concentration are disclosed. The fuel modules of the reactor core contain at least two different types of fuel elements, a high enrichment fuel element and a low enrichment fuel element. The two types of fuel elements are arranged in the fuel module with the low enrichment fuel elements located between the high moderator regions and the high enrichment fuel elements. Preferably, shim rods made of a fertile material are provided in selective regions for controlling the reactivity of the reactor by movement of the shim rods into and out of the reactor core. The moderation of neutrons adjacent the high enrichment fuel elements is preferably minimized as by reducing the spacing of the high enrichment fuel elements and/or using a moderator having a reduced moderating effect.

  19. Anaerobic oxidation of arsenite in Mono Lake water and by a facultative, arsenite-oxidizing chemoautotroph, strain MLHE-1

    Science.gov (United States)

    Oremland, R.S.; Hoeft, S.E.; Santini, J.M.; Bano, N.; Hollibaugh, R.A.; Hollibaugh, J.T.

    2002-01-01

    Arsenite [As(III)]-enriched anoxic bottom water from Mono Lake, California, produced arsenate [As(V)] during incubation with either nitrate or nitrite. No such oxidation occurred in killed controls or in live samples incubated without added nitrate or nitrite. A small amount of biological As(III) oxidation was observed in samples amended with Fe(III) chelated with nitrolotriacetic acid, although some chemical oxidation was also evident in killed controls. A pure culture, strain MLHE-1, that was capable of growth with As(III) as its electron donor and nitrate as its electron acceptor was isolated in a defined mineral salts medium. Cells were also able to grow in nitrate-mineral salts medium by using H2 or sulfide as their electron donor in lieu of As(III). Arsenite-grown cells demonstrated dark 14CO2 fixation, and PCR was used to indicate the presence of a gene encoding ribulose-1,5-biphosphate carboxylase/oxygenase. Strain MLHE-1 is a facultative chemoautotroph, able to grow with these inorganic electron donors and nitrate as its electron acceptor, but heterotrophic growth on acetate was also observed under both aerobic and anaerobic (nitrate) conditions. Phylogenetic analysis of its 16S ribosomal DNA sequence placed strain MLHE-1 within the haloalkaliphilic Ectothiorhodospira of the ??-Proteobacteria. Arsenite oxidation has never been reported for any members of this subgroup of the Proteobacteria.

  20. Linking microbial community, environmental variables and methanogenesis in anaerobic biogas digesters of chemically enhanced primary treatment sludge.

    Science.gov (United States)

    Ju, Feng; Lau, Frankie; Zhang, Tong

    2017-02-27

    Understanding the influences of biotic and abiotic factors on microbial community structure and methanogenesis are important for its engineering and ecological significance. In this study, four biogas digesters were supplied with the same inoculum and feeding sludge, but operated at different sludge retention time (7 to 16 days) and organic loading rates for 90 days to determine the relative influence of biotic and environmental factors on the microbial community assembly and methanogenic performance. Despite different operational parameters, all digester communities were dominated by Bacteroidales, Clostridiales and Thermotogales, and followed the same trend of population dynamics over time. Network and multivariate analyses suggest that deterministic factors, including microbial competition (involving Bacteroidales spp.), niche differentiation (e.g., within Clostridiales spp.), and periodic microbial immigration (from feed sludge), are the key drivers of microbial community assembly and dynamics. A yet-to-be-cultured phylotype of Bacteroidales (GenBank ID: GU389558.1) is implicated as a strong competitor for carbohydrates. Moreover, biogas-producing rate and methane content were significantly related with the abundances of functional populations rather than any operational or physicochemical parameter, revealing microbiological mediation of methanogenesis. Combined, this study enriches our understandings of biological and environmental drivers of microbial community assembly and performance in anaerobic digesters.

  1. Biomass stabilization in the anaerobic digestion of wastewater sludges

    Energy Technology Data Exchange (ETDEWEB)

    Arnaiz, C. [Universidad de Sevilla, Dept. de Ingenieria Quimica y Ambiental, Sevilla (Spain); Gutierrez, J.C. [Universidad Pablo de Olavide, Dept. de Ciencias Ambientales, Sevilla (Spain); Lebrato, J. [Universidad de Sevilla, Grupo Tratamiento de Aguas Residuales, Sevilla (Spain)

    2005-07-01

    Sludge stabilization processes include both volatile solid destruction and biomass stabilization. Traditionally, both processes have been considered together, in such a way that, when volatile solid destruction is achieved, the biomass is considered stabilized. In this study, volatile solids reduction and biomass stabilization in the anaerobic digestion of primary, secondary and mixed sludges from municipal wastewater treatment plants were researched in batch cultures by measurements of suspended solids and suspended lipid-phosphate. The estimated kinetic constants were higher in all sludge types tested for the biomass stabilization process, indicating that volatile solids destruction and biomass stabilization are not parallel processes, since the latter one is reached before the former. (Author)

  2. Biomass stabilization in the anaerobic digestion of wastewater sludges

    Energy Technology Data Exchange (ETDEWEB)

    Arnaiz, C. [Universidad de Sevilla (Spain). Escuela Universitaria Politecnica. Departamento de Ingenieria Quimica y Ambiental; Gutierrez, J.C. [Universidad Pablo de Olavide, Sevilla (Spain). Departamento de Ciencias Ambientales; Lebrato, J. [Universidad de Sevilla (Spain). Escuela Universitaria Politecnica

    2006-07-15

    Sludge stabilization processes include both volatile solid destruction and biomass stabilization. Traditionally, both processes have been considered together, in such a way that, when volatile solid destruction is achieved, the biomass is considered stabilized. In this study, volatile solids reduction and biomass stabilization in the anaerobic digestion of primary, secondary and mixed sludges from municipal wastewater treatment plants were researched in batch cultures by measurements of suspended solids and suspended lipid-phosphate. The estimated kinetic constants were higher in all sludge types tested for the biomass stabilization process, indicating that volatile solids destruction and biomass stabilization are not parallel processes, since the latter one is reached before the former. (author)

  3. Isolation and Characterization of Microbes Mediating Thermodynamically Favorable Coupling of Anaerobic Oxidation of Methane and Metal Reduction

    Science.gov (United States)

    Glass, J. B.; Reed, B. C.; Sarode, N. D.; Kretz, C. B.; Bray, M. S.; DiChristina, T. J.; Stewart, F. J.; Fowle, D. A.; Crowe, S.

    2014-12-01

    Methane is the third most reduced environmentally relevant electron donor for microbial metabolisms after organic carbon and hydrogen. In anoxic ecosystems, the major sink for methane is anaerobic oxidation of methane (AOM) mediated by syntrophic microbial consortia that couple AOM to reduction of an oxidized electron acceptor to yield free energy. In marine sediments, AOM is generally coupled to reduction of sulfate despite an extremely small amount of free energy yield because sulfate is the most abundant electron acceptor in seawater. While AOM coupled to Fe(III) and Mn(IV) reduction (Fe- and Mn-AOM) is 10-30x more thermodynamically favorable than sulfate-AOM, and geochemical data suggests that it occurs in diverse environments, the microorganisms mediating Fe- and Mn-AOM remain unknown. Lake Matano, Indonesia is an ideal ecosystem to enrich for Fe- and Mn-AOM microbes because its anoxic ferruginous deep waters and sediments contain abundant Fe(III), Mn(IV) and methane, and extremely low sulfate and nitrate. Our research aims to isolate and characterize the microbes mediating Fe- and Mn-AOM from three layers of Lake Matano sediments through serial enrichment cultures in minimal media lacking nitrate and sulfate. 16S rRNA amplicon sequencing of sediment inoculum revealed the presence of the Fe(III)-reducing bacterium Geobacter (5-10% total microbial community in shallow sediment and 35-60% in deeper sediment) as well as 1-2% Euryarchaeota implicated in methane cycling, including ANME-1 and 2d and Methanosarcinales. After 90 days of primary enrichment, all three sediment layers showed high levels of Fe(III) reduction (60-90 μM Fe(II) d-1) in the presence of methane compared to no methane and heat-killed controls. Treatments with added Fe(III) as goethite contained higher abundances of Geobacter than the inoculum (60-80% in all layers), suggesting that Geobacter may be mediating Fe(III) reduction in these enrichments. Quantification of AOM rates is underway, and

  4. Anaerobic ammonium oxidation in a bioreactor treating slaughterhouse wastewater

    Directory of Open Access Journals (Sweden)

    V. Reginatto

    2005-12-01

    Full Text Available Ammonium oxidation was thought to be an exclusively aerobic process; however, as recently described in the literature, it is also possible under anaerobic conditions and this process was named ANAMMOX. This work describes the operation of a system consisting of a denitrifying reactor coupled to a nitrifying reactor used for removal of nitrogen from slaughterhouse wastewater. During operation of the denitrifying reactor an average nitrogen ammonium removal rate of 50 mg/Ld was observed. This biomass was used to seed a second reactor, operated in repeated fed batch mode, fed with synthetic medium specific to the growth of bacteria responsible for the ANAMMOX process. The nitrogen loading rate varied between 33 and 67 mgN/Ld and average nitrogen removal was 95% and 40%, respectively. Results of fluorescence in situ hybridization (FISH confirmed the presence of anammox-like microorganisms in the enriched biomass.

  5. Life at extreme limits: the anaerobic halophilic alkalithermophiles.

    Science.gov (United States)

    Mesbah, Noha M; Wiegel, Juergen

    2008-03-01

    The ability of anaerobic microorganisms to proliferate under extreme conditions is of widespread importance for microbial physiology, remediation, industry, and evolution. The halophilic alkalithermophiles are a novel group of polyextremophiles. Tolerance to alkaline pH, elevated NaCl concentrations, and high temperatures necessitates mechanisms for cytoplasmic pH acidification; permeability control of the cell membrane; and stability of proteins, the cell wall, and other cellular constituents to multiple extreme conditions. Although it is generally assumed that extremophiles growing at more than one extreme combine adaptive mechanisms for each individual extreme, adaptations for individual extremes often counteract each other. However, in alkaline, hypersaline niches heated via intense solar irradiation, culture-independent analyses have revealed the presence of an extensive diversity of aerobic and anaerobic microorganisms belonging to Bacteria and Archaea that survive and grow under multiple harsh conditions. Thus, polyextremophiles must have developed novel adaptive strategies enabling them to grow and proliferate under multiple extreme conditions. The recent isolation of two novel anaerobic, halophilic alkalithermophiles, Natranaerobius thermophilus and Halonatronum saccharophilum, will provide a platform for detailed biochemical, genomic, and proteomic experiments, allowing a greater understanding of the novel adaptive mechanisms undoubtedly employed by polyextremophiles. In this review, we highlight growth characteristics, ecology, and phylogeny of the anaerobic halophilic alkalithermophiles isolated. We also describe the bioenergetic and physiological problems posed by growth at the multiple extreme conditions of alkaline pH, high NaCl concentration, and elevated temperature under anoxic conditions and highlight recent findings and unresolved problems regarding adaptation to multiple extreme conditions.

  6. Benzene removal by a novel modification of enhanced anaerobic biostimulation.

    Science.gov (United States)

    Xiong, Wenhui; Mathies, Chris; Bradshaw, Kris; Carlson, Trevor; Tang, Kimberley; Wang, Yi

    2012-10-01

    A novel modification of enhanced anaerobic bioremediation techniques was developed by using non-activated persulfate to accelerate the organic phosphorus breakdown and then stimulate benzene biodegradation by nitrate and sulfate reduction. Benzene concentrations in groundwater where nitrate, triethyl phosphate and persulfate were successfully injected were reduced at removal efficiencies greater than 77% to the levels below the applicable guideline. Soil benzene was removed effectively by the modification of the enhanced anaerobic bioremediation with removal efficiencies ranging between 75.9% and 92.8%. Geochemical analytical results indicated that persulfate effectively breaks down triethyl phosphate into orthophosphate, thereby promoting nitrate and sulfate utilization. Microbial analyses (quantitative polymerase chain reaction, denaturing gradient gel electrophoresis and 16S ribosomal RNA) demonstrated that benzene was primarily biodegraded by nitrate reduction while sulfate reduction played an important role in benzene removal at some portions of the study site. Enrichment in the heavier carbon isotope ¹³C of residual benzene with the increased removal efficiency provided direct evidence for benzene biodegradation. Nitrogen, sulfur and oxygen isotope analyses indicated that both nitrate reduction and sulfate reduction were occurring as bioremediation mechanisms.

  7. Thermal pretreatment of algae for anaerobic digestion.

    Science.gov (United States)

    Marsolek, Michael D; Kendall, Elizabeth; Thompson, Phillip L; Shuman, Teodora Rutar

    2014-01-01

    The objective of this work was to determine the benefit of thermal pretreatment on biogas yield from microalgae-fed anaerobic digester mesocosms. Replicate Nanochloropsis oculata cultures were heated for 4h at 30, 60, and 90°C, as well as at a constant temperature of 90°C for 1, 3.5, and 12h. Net biogas production increased from 0.28L biogas/g volatile solids added (VSa) for the control to 0.39 L biogas/g VSa (p<0.01) when heated at 90°C, but there was no improvement at 30 or 60°C. Increased biogas production correlated with increased soluble chemical oxygen demand (COD). Net biogas production increased as a function of heating time, from 0.32 L biogas/g VSa for the control, to 0.41, 0.43, and 0.44 L biogas/g VSa (p<0.05 for all combinations vs. control) when preheated at 90°C for 1, 3.5, and 12h, respectively. However, despite enhanced biogas production the energy balance is negative for thermal pretreatment.

  8. Carbon monoxide conversion by anaerobic bioreactor sludges

    NARCIS (Netherlands)

    Sipma, J.; Stams, A.J.M.; Lens, P.N.L.; Lettinga, G.

    2003-01-01

    Seven different anaerobic sludges from wastewater treatment reactors were screened for their ability to convert carbon monoxide (CO) at 30 and 55degreesC
    Seven different anaerobic sludges from wastewater treatment reactors were screened for their ability to convert carbon monoxide (CO) at 30 and

  9. Anaerobic critical velocity in four swimming techniques.

    Science.gov (United States)

    Neiva, H P; Fernandes, R J; Vilas-Boas, J P

    2011-03-01

    The aim of this study was to assess critical velocity in order to control and evaluate anaerobic swimming training. 51 highly trained male swimmers performed maximal 15, 25, 37.5 and 50 m in the 4 swimming techniques to determine critical velocity from the distance-time relationship. Anaerobic critical velocity was compared with 100 m swimming performance and corresponding partials. Complementarily, 9 swimmers performed a 6×50 m (4 min interval) training series at front crawl individual anaerobic critical velocity, capillary blood lactate concentrations being assessed after each repetition. The mean±SD values of anaerobic critical velocity and its relationship with the 100 m event were: 1.61±0.07 (r=0.60, p=0.037), 1.53±0.05 (r=0.81, p=0.015), 1.33±0.05 (r=0.83, p=0.002), and 1.75±0.05 (r=0.74, p=0.001), for butterfly, backstroke, breaststroke and front crawl, respectively. However, differences between anaerobic critical velocity and performance were observed (with exception of the second half of the 100 m swimming events in breaststroke and butterfly). Lactate concentration values at the end of the series were 14.52±1.06 mmol.l (-1), which suggests that it was indeed an anaerobic training set. In this sense, anaerobic critical velocity can be used to prescribe anaerobic training intensities.

  10. Integrated anaerobic and aerobic treatment of sewage.

    NARCIS (Netherlands)

    Kaijun Wang,

    1994-01-01

    This thesis describes results of investigations dealing with sequential concept of anaerobic-aerobic treatment of municipal wastewater. The main purposes of the study were 1) to develop a proper anaerobic hydrolytic pretreatment unit, consisting of a Hydrolysis Upflow Sludge Bed (HUSB-) reactor and

  11. Anaerobic degradation of linear alkylbenzene sulfonate

    DEFF Research Database (Denmark)

    Mogensen, Anders Skibsted; Haagensen, Frank; Ahring, Birgitte Kiær

    2003-01-01

    increases during anaerobic stabilization due to transformation of easily degradable organic matter. Hence, LAS is regarded as resistant to biodegradation under anaerobic conditions. We present data from a lab-scale semi-continuously stirred tank reactor (CSTR) spiked with linear dodecylbenzene sulfonate (C...

  12. Prospects of Anaerobic Digestion Technology in China

    Institute of Scientific and Technical Information of China (English)

    2007-01-01

    As the world's largest developing country, China must face the problem of managing municipal solid waste, and the challenge of organic waste disposal is even more serious. Considering the characteristics of traditional waste disposal technologies and the subsequent secondary pollution, anaerobic digestion has various advantages such as reduction in the land needed for disposal and preservation of environmental quality. In light of the energy crisis, this paper focuses on the potential production of biogas from biowaste through anaerobic digestion processes, the problems incurred by the waste collection system, and the efficiency of the anaerobic digestion process. Use of biogas in a combined heat and power cogeneration system is also discussed. Finally, the advantages of anaerobic digestion technology for the Chinese market are summarized. The anaerobic digestion is suggested to be a promising treating technology for the organic wastes in China.

  13. Toxicants inhibiting anaerobic digestion: a review.

    Science.gov (United States)

    Chen, Jian Lin; Ortiz, Raphael; Steele, Terry W J; Stuckey, David C

    2014-12-01

    Anaerobic digestion is increasingly being used to treat wastes from many sources because of its manifold advantages over aerobic treatment, e.g. low sludge production and low energy requirements. However, anaerobic digestion is sensitive to toxicants, and a wide range of compounds can inhibit the process and cause upset or failure. Substantial research has been carried out over the years to identify specific inhibitors/toxicants, and their mechanism of toxicity in anaerobic digestion. In this review we present a detailed and critical summary of research on the inhibition of anaerobic processes by specific organic toxicants (e.g., chlorophenols, halogenated aliphatics and long chain fatty acids), inorganic toxicants (e.g., ammonia, sulfide and heavy metals) and in particular, nanomaterials, focusing on the mechanism of their inhibition/toxicity. A better understanding of the fundamental mechanisms behind inhibition/toxicity will enhance the wider application of anaerobic digestion.

  14. Startup of anaerobic fluidized bed reactors with acetic acid as the substrate.

    Science.gov (United States)

    Hsu, Y; Shieh, W K

    1993-02-05

    The startup of anaerobic fluidized bed reactors, which use Manville R-633 beads as the growth support media, acetate enriched bacterial culture as the inoculum, and acetic acid as the sole substrate, is studied. Tow startup strategies are evaluated: one based on maximum and stable substrate utilization and another based on maximum substrate loading controlled by reactor pH. The startup process is characterized using a number of operational parameters.The reactors again excellent total organic carbon (TOC) removal (i.e., > 97% at a feed concentration of 5000 mg TOC/L) and stable methane production (i.e., 0.90 L CH(4)/g TOC, where TOC(r) is TOC removed) at a early stage of the startup process, regardless of the strategies applied. The loading can be increased rapidly without the danger of being overloaded. Significant losses of growth support media and biomass caused by gas effervescence at higher loadings limits the maximum loading that can be safely applied during startup process.A high reactor immobilized biomass inventory is achievable using the porous growth support media (e.g., Manville 633 beads). A rapid increase in loading creates a substrate rich environment that yields more viable reactor biomass. Both substrate utilization rate (batch and continuous) and immobilized biomass inventory stabilize concomitantly at the late stage of the startup process, indicating the attainment of steady-state conditions in reactors. Therefore, they are better parameters that TOC removal and methane production for characterizing the entire startup process of aerobic fluidized bed reactor.The strategy based on maximum substrate loading controlled by reactor pH significantly shortens the startup time. In this case, the reactor attains steady-state conditions approximately 140 days after startup. On the other hand, a startup time of 200 days is required when the strategy based maximum substrate utilization is adopted.

  15. Effect of interactions between carbon dioxide enrichment and NH4+/NO3- ratio on pH of culturing nutrient solution,growth and vigor of tomato root system

    Institute of Scientific and Technical Information of China (English)

    Juan LI; Jianmin ZHOU

    2008-01-01

    A growth chamber experiment was conducted to investigate the influence of NH4+/NO3- ratio and elevated CO2 concentration on the pH in nutrient solution,growth and root vigor system of tomato seedling roots,which attempts to understand whether the elevated CO2 concentration can alleviate the harmful effects of higher NH4+-N concentration in nutrient solutions on the tomato root system.Tomato (Lycopersicon esculenturn Mill.var.Hezuo 906) was grown in pots with nutrient solutions varying in NH4+/NO3- ratio (0:1,1:3,1:1,3:1and 1:0) and the growth chambers were supplied with with the growth process and CO2 concentration increased.At both CO2 levels,pH increased when 100% NO3--N was supplied and decreased in other treatments.The pH decrease in the nutrient solution was directly correlated to the NH4+-N proportion.The pH value was more reduced in 100% NH4+-N nutrient solution than increased in the 100% NO3--N nutrient solution.CO2 enrichment increased the dry weight of shoots and roots,root vigor system,total absorbing area and active absorbing area of tomato seedlings.All the measurement indexes above were increased in the elevated CO2 concentration treatment with the NO3- proportion increase in the nutrient solutions.Thus,under the elevated CO2 concentration,the dry weights of shoots and roots,root vigor system,total root absorbing area and active absorbing area were found to be inversely correlated to NH4+/NO3- ratio,leading to about 65.8%,78.0%,18.9%,12.9% and 18.9% increase,respectively,compared with that under the ambient CO2 concentration.Our results indicated that tomato seedling roots may benefit mostly from CO2 enrichment when 100% NO3--N nutrient solutions was supplied,but the CO2 concentration elevation did not alleviate the harmful effects when 100% NHa+-N was supplied.

  16. Bacteremia due to anaerobic bacteria: epidemiology in a northern Bari Hospital, Italy

    Directory of Open Access Journals (Sweden)

    Maria Antonietta Distasi

    2015-06-01

    Full Text Available Background. Anaerobic bacteria are part of the commensal bacterial flora of skin and mucosae. Iatrogenic and pathological conditions altering this commensal relationship cause life-threatening diseases. Materials and Methods. We analysed the blood cultures sent to the microbiology of our hospital between 2008 and the first quarter of 2013 to measure the frequency of bacteraemia caused by anaerobia. We examined 3138 vials of blood cultures for anaerobia, inoculated following in-house standard procedures. The colonies grown in absence of air were subjected to biochemical analysis. The MICs of metronidazole for 23 of the 26 organisms was tested. Results. Twelve bacteria of the Bacteroides genus were identified, 9 Propionibacterium acnes, 1 Peptosctreptococcus micros, 1 Lactobacillus acidophilus, 1 Clostridium perfringens, 1 Prevotella oralis, 1 Eubacterium lentum. Conclusions. The analysis of the results suggests that the incidence of cultures positive to anaerobia was constant across the years. We note that advanced age, altered mucocutaneous tropism, alterations to the oral and intestinal bacterial flora intensify the risk of anaerobial pathogenicity. The analysis of the metronidazole-determined MIC suggests that the intestinal anaerobic flora responds well to therapy and prophylaxis with Metronidazole, while the anaerobic bacteria residing on skin and other mucosae are resistant. It is however hard to determine the clinical impact of anaerobic bacteremiae and their effect on the outcome of the patient, due to the scarcity of available clinical data.

  17. Anaerobic Bacteremia: Impact of Inappropriate Therapy on Mortality

    Science.gov (United States)

    Lee, Yangsoon; Park, Yongjung; Kim, Myungsook; Choi, Jun Yong; Yong, Dongeun; Jeong, Seok Hoon

    2016-01-01

    Background Investigation on incidence and mortality of anaerobic bacteremia (AB) is clinically relevant in spite of its infrequent occurrence and not often explored, which report varies according to period and institutions. Therefore, it is necessary to analyze the incidence and risk factors related to mortality and assess clinical outcomes of AB in current aspect. Materials and Methods Characteristics of AB patients and anaerobic bacteria from blood culture at a university hospital in 2012 were reviewed retrospectively. The correlation between risk factors and 28-day patient mortality was analyzed. Results A total of 70 non-duplicated anaerobic bacteria were isolated from blood of 70 bacteremia patients in 2012. The history of cardiovascular disease as host's risk factor was statistically significant (P = 0.0344) in univariate and multivariate analysis. Although the inappropriate therapy was not statistically significant in univariate and multivariate analysis, the survival rate of bacteremia was significantly worse in patients who had inappropriate therapy compared with those underwent appropriate therapy (hazard ratio, 5.4; 95% confidence interval, 1.7–6.9; P = 0.004). The most frequently isolated organism was Bacteroides fragilis (32 isolates, 46%), followed by Bacteroides thetaiotaomicron (10, 14%), and non-perfringens Clostridium (7, 10%). Conclusion The incidence of AB in 2012 was 2.3% (number of AB patients per 100 positive blood culture patients) and the mortality rate in patients with clinically significant AB was 21.4%. In addition, AB was frequently noted in patients having malignancy and the survival rate of AB was significantly worse in patients who received inappropriate therapy compared with those underwent appropriate therapy. PMID:27433379

  18. 构建一种高产量小胶质细胞体外纯化培养的方法%A new method for culturing highly enriched microglia in vitro

    Institute of Scientific and Technical Information of China (English)

    魏桂荣; 张敏; 董继华; 梅元武; 刘仁刚

    2005-01-01

    BACKGROUND: Microglial cells are prominently involved in certain neurologic diseases such as Parkinson disease and Alzeheimer disease. In vitro primary culture is commonly used in studies on the functions of microglia.However, these classical culture methods have some defects including complex procedures and low out-put.OBJECTIVE: To establish a simplified high-output primary culture of microglia.DESIGN: An explorative experiment with microglial cells as the single sample.SETTING: Department of Neurology, Affiliated Union Hospital, Tongji Medical College, Huazhong University of Science and Technology.MATERIALS: The study was finished at the Central Laboratory of Union Hospital from April to October 2004. Microglial cells were obtained from 10 newborn(one day) male Kunming mice that were selected.METHODS: The author' s culture method was based on McCarthy method, we developed a new culture method and made some improvements,including the increased cell density for primary culture and nutritional deprivation. The microlglial cells were isolated with low-concentration trypsin-EDTA(ethylene diamine tetraacetic acid) digestion and immunochemically labeled with MAC-1 antibody, so as to measure the output and purity of microglia.MAIN OUTCOME MEASURES: ① Morphologic features of microglial cells, observed with inverted microscope; ② Purity and activity of microglia cultured with these two methods, were measured immunohistochemically.RESULTS: For microglia cultured with McCarthy method, the culture cycle was 20 days and the output was 2 × l05 cells per flask with a purity of 95% -97%. The new method shortened the culture cycle to 15 days and the output reached 1 × 106 cells per flask with a purity of 96-98%. Cell purity and activity had no significant difference between these two culture methods.CONCLUSION: The new method has a similar purity and activity with classical method; however, it may simplify procedures, shorten cycle, and increase output, and therefore can be

  19. Hydrogen-enriched fuels

    Energy Technology Data Exchange (ETDEWEB)

    Roser, R. [NRG Technologies, Inc., Reno, NV (United States)

    1998-08-01

    NRG Technologies, Inc. is attempting to develop hardware and infrastructure that will allow mixtures of hydrogen and conventional fuels to become viable alternatives to conventional fuels alone. This commercialization can be successful if the authors are able to achieve exhaust emission levels of less than 0.03 g/kw-hr NOx and CO; and 0.15 g/kw-hr NMHC at full engine power without the use of exhaust catalysts. The major barriers to achieving these goals are that the lean burn regimes required to meet exhaust emissions goals reduce engine output substantially and tend to exhibit higher-than-normal total hydrocarbon emissions. Also, hydrogen addition to conventional fuels increases fuel cost, and reduces both vehicle range and engine output power. Maintaining low emissions during transient driving cycles has not been demonstrated. A three year test plan has been developed to perform the investigations into the issues described above. During this initial year of funding research has progressed in the following areas: (a) a cost effective single-cylinder research platform was constructed; (b) exhaust gas speciation was performed to characterize the nature of hydrocarbon emissions from hydrogen-enriched natural gas fuels; (c) three H{sub 2}/CH{sub 4} fuel compositions were analyzed using spark timing and equivalence ratio sweeping procedures and finally; (d) a full size pick-up truck platform was converted to run on HCNG fuels. The testing performed in year one of the three year plan represents a baseline from which to assess options for overcoming the stated barriers to success.

  20. CLEAN: CLustering Enrichment ANalysis

    Directory of Open Access Journals (Sweden)

    Medvedovic Mario

    2009-07-01

    Full Text Available Abstract Background Integration of biological knowledge encoded in various lists of functionally related genes has become one of the most important aspects of analyzing genome-wide functional genomics data. In the context of cluster analysis, functional coherence of clusters established through such analyses have been used to identify biologically meaningful clusters, compare clustering algorithms and identify biological pathways associated with the biological process under investigation. Results We developed a computational framework for analytically and visually integrating knowledge-based functional categories with the cluster analysis of genomics data. The framework is based on the simple, conceptually appealing, and biologically interpretable gene-specific functional coherence score (CLEAN score. The score is derived by correlating the clustering structure as a whole with functional categories of interest. We directly demonstrate that integrating biological knowledge in this way improves the reproducibility of conclusions derived from cluster analysis. The CLEAN score differentiates between the levels of functional coherence for genes within the same cluster based on their membership in enriched functional categories. We show that this aspect results in higher reproducibility across independent datasets and produces more informative genes for distinguishing different sample types than the scores based on the traditional cluster-wide analysis. We also demonstrate the utility of the CLEAN framework in comparing clusterings produced by different algorithms. CLEAN was implemented as an add-on R package and can be downloaded at http://Clusteranalysis.org. The package integrates routines for calculating gene specific functional coherence scores and the open source interactive Java-based viewer Functional TreeView (FTreeView. Conclusion Our results indicate that using the gene-specific functional coherence score improves the reproducibility of the

  1. Microbial decolorization of reactive black-5 in a two-stage anaerobic-aerobic reactor using acclimatized activated textile sludge.

    Science.gov (United States)

    Mohanty, Sagarika; Dafale, Nishant; Rao, Nageswara Neti

    2006-10-01

    A two-stage anaerobic-aerobic treatment process based on mixed culture of bacteria isolated from textile dye effluent was used to degrade reactive black 5 dye (RB-5). The anaerobic step was studied in more detail by varying the dye concentration from 100 to 3000 mg l(-1). The results showed that major decolorization was achieved during the anaerobic process. The time required for decolorization by > 90% increased as the concentration of the dye increased. It was also found that maintaining dissolved oxygen (DO) concentration below 0.5 mg l(-1 )and addition of a co-substrate viz., glucose, facilitates anaerobic decolorization reaction remarkably. An attempt was made to identify the metabolites formed in anaerobic process by using high performance liquid chromatography (HPLC) and UV-VIS spectrophotometry. A plate assay was performed for the detection of dominant decolorizing bacteria. Only a few bacterial colonies with high clearing zones (decolorization zones) were found. The results showed that under anaerobic condition RB-5 molecules were reduced and aromatic amines were generated. The aromatic amine metabolite was partly removed in subsequent aerobic bio-treatment. It was possible to achieve more than 90% decolorization and approximately 46% reduction in amine metabolite concentration through two-stage anaerobic-aerobic treatment after a reaction period of 2 days.

  2. In vitro culture of previously uncultured oral bacterial phylotypes.

    Science.gov (United States)

    Thompson, Hayley; Rybalka, Alexandra; Moazzez, Rebecca; Dewhirst, Floyd E; Wade, William G

    2015-12-01

    Around a third of oral bacteria cannot be grown using conventional bacteriological culture media. Community profiling targeting 16S rRNA and shotgun metagenomics methods have proved valuable in revealing the complexity of the oral bacterial community. Studies investigating the role of oral bacteria in health and disease require phenotypic characterizations that are possible only with live cultures. The aim of this study was to develop novel culture media and use an in vitro biofilm model to culture previously uncultured oral bacteria. Subgingival plaque samples collected from subjects with periodontitis were cultured on complex mucin-containing agar plates supplemented with proteose peptone (PPA), beef extract (BEA), or Gelysate (GA) as well as on fastidious anaerobe agar plus 5% horse blood (FAA). In vitro biofilms inoculated with the subgingival plaque samples and proteose peptone broth (PPB) as the growth medium were established using the Calgary biofilm device. Specific PCR primers were designed and validated for the previously uncultivated oral taxa Bacteroidetes bacteria HOT 365 and HOT 281, Lachnospiraceae bacteria HOT 100 and HOT 500, and Clostridiales bacterium HOT 093. All agar media were able to support the growth of 10 reference strains of oral bacteria. One previously uncultivated phylotype, Actinomyces sp. HOT 525, was cultivated on FAA. Of 93 previously uncultivated phylotypes found in the inocula, 26 were detected in in vitro-cultivated biofilms. Lachnospiraceae bacterium HOT 500 was successfully cultured from biofilm material harvested from PPA plates in coculture with Parvimonas micra or Veillonella dispar/parvula after colony hybridization-directed enrichment. The establishment of in vitro biofilms from oral inocula enables the cultivation of previously uncultured oral bacteria and provides source material for isolation in coculture.

  3. Medium factors on anaerobic production of rhamnolipids by Pseudomonas aeruginosa SG and a simplifying medium for in situ microbial enhanced oil recovery applications.

    Science.gov (United States)

    Zhao, Feng; Zhou, Jidong; Han, Siqin; Ma, Fang; Zhang, Ying; Zhang, Jie

    2016-04-01

    Aerobic production of rhamnolipid by Pseudomonas aeruginosa was extensively studied. But effect of medium composition on anaerobic production of rhamnolipid by P. aeruginosa was unknown. A simplifying medium facilitating anaerobic production of rhamnolipid is urgently needed for in situ microbial enhanced oil recovery (MEOR). Medium factors affecting anaerobic production of rhamnolipid were investigated using P. aeruginosa SG (Genbank accession number KJ995745). Medium composition for anaerobic production of rhamnolipid by P. aeruginosa is different from that for aerobic production of rhamnolipid. Both hydrophobic substrate and organic nitrogen inhibited rhamnolipid production under anaerobic conditions. Glycerol and nitrate were the best carbon and nitrogen source. The commonly used N limitation under aerobic conditions was not conducive to rhamnolipid production under anaerobic conditions because the initial cell growth demanded enough nitrate for anaerobic respiration. But rhamnolipid was also fast accumulated under nitrogen starvation conditions. Sufficient phosphate was needed for anaerobic production of rhamnolipid. SO4(2-) and Mg(2+) are required for anaerobic production of rhamnolipid. Results will contribute to isolation bacteria strains which can anaerobically produce rhamnolipid and medium optimization for anaerobic production of rhamnolipid. Based on medium optimization by response surface methodology and ions composition of reservoir formation water, a simplifying medium containing 70.3 g/l glycerol, 5.25 g/l NaNO3, 5.49 g/l KH2PO4, 6.9 g/l K2HPO4·3H2O and 0.40 g/l MgSO4 was designed. Using the simplifying medium, 630 mg/l of rhamnolipid was produced by SG, and the anaerobic culture emulsified crude oil to EI24 = 82.5 %. The simplifying medium was promising for in situ MEOR applications.

  4. Impact of acclimation methods on microbial communities and performance of anaerobic fluidized bed membrane bioreactors

    KAUST Repository

    Labarge, Nicole

    2016-10-17

    An anaerobic fluidized bed membrane bioreactor (AFMBR) is a new and effective method for energy-efficient treatment of low strength wastewater, but the factors that affect performance are not well known. Different inocula and acclimation methods of the granular activated carbon (GAC) used in the reactor were examined here to determine their impact on chemical oxygen demand (COD) removal and microbial community composition of domestic wastewater-fed AFMBRs. AFMBRs inoculated with anaerobic digester sludge (D) or domestic wastewater (W) and fed domestic wastewater, or inoculated with a microbiologically diverse anaerobic bog sediment and acclimated using methanol (M), all produced the same COD removal of 63 ± 12% using a diluted wastewater feed (100 ± 21 mg L−1 COD). However, an AFMBR with GAC inoculated with anaerobic digester sludge and acclimated using acetate (A) showed significantly increased wastewater COD removal to 84 ± 6%. In addition, feeding the AFMBR with the M-acclimated GAC with an acetate medium for one week subsequently increased COD removal to 70 ± 6%. Microbial communities enriched on the GAC included Geobacter, sulfur-reducing bacteria, Syntrophaceae, and Chlorobiaceae, with reactor A having the highest relative abundance of Geobacter. These results showed that acetate was the most useful substrate for acclimation of GAC communities, and GAC harbors unique communities relative to those in the AFMBR influent and recirculated solution.

  5. Aerobic versus Anaerobic Microbial Degradation of Clothianidin under Simulated California Rice Field Conditions.

    Science.gov (United States)

    Mulligan, Rebecca A; Tomco, Patrick L; Howard, Megan W; Schempp, Tabitha T; Stewart, Davis J; Stacey, Phillip M; Ball, David B; Tjeerdema, Ronald S

    2016-09-28

    Microbial degradation of clothianidin was characterized under aerobic and anaerobic California rice field conditions. Rate constants (k) and half-lives (DT50) were determined for aerobic and anaerobic microcosms, and an enrichment experiment was performed at various nutrient conditions and pesticide concentrations. Temperature effects on anaerobic degradation rates were determined at 22 ± 2 and 35 ± 2 °C. Microbial growth was assessed in the presence of various pesticide concentrations, and distinct colonies were isolated and identified. Slow aerobic degradation was observed, but anaerobic degradation occurred rapidly at both 25 and 35 °C. Transformation rates and DT50 values in flooded soil at 35 ± 2 °C (k = -7.16 × 10(-2) ± 3.08 × 10(-3) day(-1), DT50 = 9.7 days) were significantly faster than in 25 ± 2 °C microcosms (k= -2.45 × 10(-2) ± 1.59 × 10(-3) day(-1), DT50 = 28.3 days). At the field scale, biodegradation of clothianidin will vary with extent of oxygenation.

  6. Removal of anaerobic soluble microbial products in a biological activated carbon reactor.

    Science.gov (United States)

    Dong, Xiaojing; Zhou, Weili; He, Shengbing

    2013-09-01

    The soluble microbial products (SMP) in the biological treatment effluent are generally of great amount and are poorly biodegradable. Focusing on the biodegradation of anaerobic SMP, the biological activated carbon (BAC) was introduced into the anaerobic system. The experiments were conducted in two identical lab-scale up-flow anaerobic sludge blanket (UASB) reactors. The high strength organics were degraded in the first UASB reactor (UASB1) and the second UASB (UASB2, i.e., BAC) functioned as a polishing step to remove SMP produced in UASB1. The results showed that 90% of the SMP could be removed before granular activated carbon was saturated. After the saturation, the SMP removal decreased to 60% on the average. Analysis of granular activated carbon adsorption revealed that the main role of SMP removal in BAC reactor was biodegradation. A strain of SMP-degrading bacteria, which was found highly similar to Klebsiella sp., was isolated, enriched and inoculated back to the BAC reactor. When the influent chemical oxygen demand (COD) was 10,000 mg/L and the organic loading rate achieved 10 kg COD/(m3 x day), the effluent from the BAC reactor could meet the discharge standard without further treatment. Anaerobic BAC reactor inoculated with the isolated Klebsiella was proved to be an effective, cheap and easy technical treatment approach for the removal of SMP in the treatment of easily-degradable wastewater with COD lower than 10,000 mg/L.

  7. Riboflavin Reduces Pro-Inflammatory Activation of Adipocyte-Macrophage Co-culture. Potential Application of Vitamin B2 Enrichment for Attenuation of Insulin Resistance and Metabolic Syndrome Development

    Directory of Open Access Journals (Sweden)

    Agnieszka Irena Mazur-Bialy

    2016-12-01

    Full Text Available Due to the progressive increase in the incidence of obese and overweight individuals, cardiometabolic syndrome has become a worldwide pandemic in recent years. Given the immunomodulatory properties of riboflavin, the current study was performed to investigate the potency of riboflavin in reducing obesity-related inflammation, which is the main cause of insulin resistance, diabetes mellitus 2 or arteriosclerosis. We determined whether pretreatment with a low dose of riboflavin (10.4–1000 nM affected the pro-inflammatory activity of adipocyte-macrophage co-culture (3T3 L1-RAW 264.7 following lipopolysaccharide stimulation (LPS; 100 ng/mL which mimics obesity-related inflammation. The apoptosis of adipocytes and macrophages as well as tumor necrosis factor-alpha (TNF-α, interleukin 6 (IL-6, interleukin 1beta (IL-1β, monocyte chemotactic protein 1 (MCP-1, high-mobility group box 1 (HMGB1, transforming growth factor–beta 1 (TGFβ, interleukin 10 (IL-10, inducible nitric oxide synthase (iNOS, nitric oxide (NO, matrix metalloproteinase 9 (MMP-9, tissue inhibitor of metalloproteinases-1 (TIMP-1 expression and release, macrophage migration and adipokines (adiponectin and leptin were determined. Our results indicated an efficient reduction in pro-inflammatory factors (TNFα, IL-6, MCP-1, HMGB1 upon culture with riboflavin supplementation (500–1000 nM, accompanied by elevation in anti-inflammatory adiponectin and IL-10. Moreover, macrophage migration was reduced by the attenuation of chemotactic MCP-1 release and degradation of the extracellular matrix by MMP-9. In conclusion, riboflavin effectively inhibits the pro-inflammatory activity of adipocyte and macrophage co-cultures, and therefore we can assume that its supplementation may reduce the likelihood of conditions associated with the mild inflammation linked to obesity.

  8. Kinetics of anaerobic degradation of glycol-based type I aircraft deicing fluids.

    Science.gov (United States)

    Schoenberg, T; Veltman, S; Switzenbaum, M

    2001-01-01

    The kinetics of anaerobic degradation of glycol-based Type I aircraft deicing fluids (ADFs) were characterized using suspended-growth fill-and-draw reactors. Both Type I ADFs tested showed near-complete anaerobic degradability. First-order degradation rate constants of 3.5 d(-1) for the propylene glycol-based Type I ADF and 5.2 d(-1) for the ethylene glycol-based Type I ADF were obtained through continuous-culture means under mesophilic conditions (35 degrees C). Fill-and-draw operation at lower temperatures affected anaerobic degradability only minimally down to 25 degrees C but substantially below 25 degrees C. High Type I ADF feed concentrations substantially affected degradability. Batch testing of fill-and-draw reactors resulted in first-order degradation rate constants of 1.9 d(-1) for propylene glycol-based Type I ADF and 3.5 d(-1) for ethylene glycol-based Type I ADF.

  9. Biogas production from brewery spent grain enhanced by bioaugmentation with hydrolytic anaerobic bacteria.

    Science.gov (United States)

    Čater, Maša; Fanedl, Lijana; Malovrh, Špela; Logar, Romana Marinšek

    2015-06-01

    Lignocellulosic substrates are widely available but not easily applied in biogas production due to their poor anaerobic degradation. The effect of bioaugmentation by anaerobic hydrolytic bacteria on biogas production was determined by the biochemical methane potential assay. Microbial biomass from full scale upflow anaerobic sludge blanket reactor treating brewery wastewater was a source of active microorganisms and brewery spent grain a model lignocellulosic substrate. Ruminococcus flavefaciens 007C, Pseudobutyrivibrio xylanivorans Mz5(T), Fibrobacter succinogenes S85 and Clostridium cellulovorans as pure and mixed cultures were used to enhance the lignocellulose degradation and elevate the biogas production. P. xylanivorans Mz5(T) was the most successful in elevating methane production (+17.8%), followed by the coculture of P. xylanivorans Mz5(T) and F. succinogenes S85 (+6.9%) and the coculture of C. cellulovorans and F. succinogenes S85 (+4.9%). Changes in microbial community structure were detected by fingerprinting techniques.

  10. Enriching distinctive microbial communities from marine sediments via an electrochemical-sulfide-oxidizing process on carbon electrodes

    Directory of Open Access Journals (Sweden)

    Shiue-Lin eLi

    2015-02-01

    Full Text Available Sulfide is a common product of marine anaerobic respiration, and a potent reactant biologically and geochemically. Here we demonstrate the impact on microbial communities with the removal of sulfide via electrochemical methods. The use of differential pulse voltammetry revealed that the oxidation of soluble sulfide was seen at + mV (vs. SHE at all pH ranges tested (from pH = 4 to 8, while non-ionized sulfide, which dominated at pH = 4 was poorly oxidized via this process. Two mixed cultures (CAT and LA were enriched from two different marine sediments (from Catalina Island, CAT; from the Port of Los Angeles, LA in serum bottles using a seawater medium supplemented with lactate, sulfate, and yeast extract, to obtain abundant biomass. Both CAT and LA cultures were inoculated in electrochemical cells (using yeast-extract-free seawater medium as an electrolyte equipped with carbon-felt electrodes. In both cases, when potentials of +630 or 130 mV (vs. SHE were applied, currents were consistently higher at +630 then at 0 mV, indicating more sulfide being oxidized at the higher potential. In addition, higher organic-acid and sulfate conversion rates were found at +630 mV with CAT, while no significant differences were found with LA at different potentials. The results of microbial-community analyses revealed a decrease in diversity for both CAT and LA after electrochemical incubation. In addition, some bacteria (e.g., Clostridium and Arcobacter not well known to be capable of extracellular electron transfer, were found to be dominant in the electrochemical cells. Thus, even though the different mixed cultures have different tolerances for sulfide, electrochemical-sulfide removal can lead to major population changes.

  11. A Contrastive Research on American-Chinese Cultural Identity from Perspective of Mass Culture

    Institute of Scientific and Technical Information of China (English)

    赵欣

    2012-01-01

    The appearance and development of "Super Girl" accompanies with the rise of mass culture and it reflects the main features of mass culture.Meanwhile western mass cultural product especially the American mass cultural product "American Idol" influenced Chinese mass culture and it has produced massive impact on the identity of national cultural and cultural independence in China.The thesis can help to deepen and make up for the deficiency of mass culture theories,to enrich the applicational experience and the...

  12. Recovery of anaerobic digestion after exposure to toxicants. Final report

    Energy Technology Data Exchange (ETDEWEB)

    Yang, J.; Parkin, G.F.; Speece, R.E.

    1979-12-01

    The concept that methane fermentation cannot tolerate chronic or slug doses of toxicants has almost totally precluded methane fermentation as a viable contender for the treatment of industrial wastewaters. This study assayed a wide variety of toxicants, heavy metals, inorganic salts, organic chemicals, solvents, and antibiotics which are used in industrial processes and, therefore, appear in the industrial wastewaters therefrom. Toxicity was related to the reduction in methane production of a control containing no toxicant. The response of methane fermentation after exposure to a toxicant was assayed with unacclimated cultures as well as cultures which had been acclimated to increasing concentrations of the toxicant over long periods of time. The reversible nature of the toxicants was assayed by adding slug doses to plug flow anaerobic filters and recording gas production prior to, during, and after toxicant addition.

  13. Effects of heat shocks on microbial community structure and microbial activity of a methanogenic enrichment degrading benzoate.

    Science.gov (United States)

    Mei, R; Narihiro, T; Nobu, M K; Liu, W-T

    2016-11-01

    In anaerobic digesters, temperature fluctuation could lead to process instability and failure. It is still not well understood how digester microbiota as a whole respond to heat shock, and what specific organisms are vulnerable to perturbation or responsible for process recovery after perturbation. To address these questions, a mesophilic benzoate-degrading methanogenic culture enriched from digester was subjected to different levels of heat shock. Three types of methane production profiles after perturbation were observed in comparison to the control: uninhibited, inhibited with later recovery, and inhibited without recovery. These responses were correlated with the microbial community compositions based on the analyses of 16S rRNA and 16S rRNA gene. Specifically, the primary benzoate-degrading syntroph was highly affected by heat shock, and its abundance and activity were both crucial to the restoration of benzoate degradation after heat shock. In contrast, methanogens were stable regardless whether methane production was inhibited. Populations related to 'Candidatus Cloacimonetes' and Firmicutes showed stimulated growth. These observations indicated distinct physiological traits and ecological niches associated with individual microbial groups. The results obtained after exposure to heat shock can be critical to more comprehensive characterization of digester ecology under perturbations.

  14. Intraspecific variation in aerobic and anaerobic locomotion

    DEFF Research Database (Denmark)

    Svendsen, Jon Christian; Tirsgård, Bjørn; Cordero, Gerardo A.;

    2015-01-01

    Intraspecific variation and trade-off in aerobic and anaerobic traits remain poorly understood in aquatic locomotion. Using gilthead sea bream (Sparus aurata) and Trinidadian guppy (Poecilia reticulata), both axial swimmers, this study tested four hypotheses: (1) gait transition from steady...... to unsteady (i.e., burst-assisted) swimming is associated with anaerobic metabolism evidenced as excess post exercise oxygen consumption (EPOC); (2) variation in swimming performance (critical swimming speed; U crit) correlates with metabolic scope (MS) or anaerobic capacity (i.e., maximum EPOC); (3...

  15. Energy from anaerobic methane production. [Sweden

    Energy Technology Data Exchange (ETDEWEB)

    1982-02-01

    Since 1970 Swedish researchers have been testing the ANAMET (anaerobic-aerobic-methane) process, which involves converting industrial wastewaters via an initial anaerobic microbiological step followed by an aerobic one. Recycling the biomass material in each step allows shorter hydraulic retention times without decreasing stability or solids reduction. Since the first ANAMET plants began operating at a Swedish sugar factory in 1972, 17 more plants have started up or are under construction. Moreover, the ANAMET process has engendered to offshoot BIOMET (biomass-methane) process, a thermophilic anaerobic scheme that can handle sugar-beet pulp as well as grass and other soft, fast-growing biomasses.

  16. Anaerobic degradation and toxicity of commercial cationic surfactants in anaerobic screening tests.

    Science.gov (United States)

    García, M T; Campos, E; Sánchez-Leal, J; Ribosa, I

    2000-09-01

    Anaerobic biodegradability and toxicity on anaerobic bacteria of di(hydrogenated tallow) dimethyl ammonium chloride (DHTDMAC) and two esterquats have been investigated. A batch test system containing municipal digester solids as a source of anaerobic bacteria, based on the method proposed by the ECETOC, has been applied. To evaluate the potential toxicity of such surfactants on anaerobic sludge, a co-substrate, an easily biodegradable compound in anaerobic conditions, has been added to the samples to test and the effects on biogas production have been determined. For the esterquats studied high biodegradation levels were obtained and no toxic effects on anaerobic bacteria were observed even at the highest concentrations tested, 100 and 200 mg C/l, respectively. On the contrary, DHTDMAC was not degradated at the same test conditions. However, no inhibitory effects on the biogas production were detected for this surfactant at concentrations <100 mg C/l.

  17. Assessment of the ability of sludge to degrade PCP under anaerobic conditions

    Directory of Open Access Journals (Sweden)

    R. M. L. Bolaños

    2005-12-01

    Full Text Available The capacity of sludge from different sources to degrade pentachlorophenol (PCP was evaluated. Three 2.5 liter reactors (R1, R2, and R3 were inoculated with different anaerobic sludges, semi continuously fed and maintained in orbital motion at 30±1°C. R1 was inoculated with aerobic sludge and river sediment collected downstream from a pulp and paper plant. R2 received sludge from an anaerobic reactor treating effluents from a paper recycling plant and R3 received anaerobic sludge from a biodigestor treating industrial and domestic effluents. The sludges were first acclimatized to a culture medium generally recommended for organochloride anaerobic degradation studies. The reactors were then subjected to increasing concentrations of PCP from 0.05 to 10.0 mg.l-1. PCP degradation and metabolite formation were monitored using gas chromatography, and the effects of PCP on the anaerobic process were verified by monitoring pH, volatile fatty acids, alkalinity, total suspended solids, and chemical oxygen demand. It was found that PCP did not affect reactor performance. All the sludges displayed the best PCP degradation capacity at a concentration of 0.2 mg.l-1, producing fewer chlorinated metabolites than when higher PCP concentrations were applied. R1 consistently produced fewer chlorinated metabolites, confirming the hypothesis that pre exposure to chlorinated compounds improves the sludge's capacity to degrade PCP.

  18. Clinically significant anaerobic bacteria isolated from patients in a South African academic hospital: antimicrobial susceptibility testing.

    Science.gov (United States)

    Naidoo, S; Perovic, O; Richards, G A; Duse, A G

    2011-09-27

    BACKGROUND. Increasing resistance to some antimicrobial agents among anaerobic bacteria has made susceptibility patterns less predictable. METHOD. This was a prospective study of the susceptibility data of anaerobic organisms isolated from clinical specimens from patients with suspected anaerobic infections from June 2005 until February 2007. Specimens were submitted to the microbiology laboratory at Charlotte Maxeke Johannesburg Academic Hospital, where microscopy, culture and susceptibility testing were performed the using E test® strip minimum inhibitory concentration method. Results were interpreted with reference to Clinical and Laboratory Standards Institute guidelines for amoxicillin-clavulanate, clindamycin, metronidazole, penicillin, ertapenem, cefoxitin, ceftriaxone, chloramphenicol and piperacillin-tazobactam. RESULTS. One hundred and eighty anaerobic isolates were submitted from 165 patients. The most active antimicrobial agents were chloramphenicol (100% susceptible), ertapenem (97.2%), piperacillin-tazobactam (99.4%) and amoxicillin-clavulanic acid (96.7%). Less active were metronidazole (89.4%), cefoxitin (85%), clindamycin (81.7%), ceftriaxone (68.3%) and penicillin (33.3%). CONCLUSION. Susceptibility testing should be performed periodically to identify emerging trends in resistance and to modify empirical treatment of anaerobic infections.

  19. Microbial populations of an upflow anaerobic sludge blanket reactor treating wastewater from a gelatin industry.

    Science.gov (United States)

    Vieira, A M; Bergamasco, R; Gimenes, M L; Nakamura, C V; Dias Filho, B P

    2001-12-01

    The microbial populations of an upflow anaerobic sludge blanket reactor, used for treating wastewater from the gelatin industry, were studied by microbiological methods and phase-contrast and electron microscopy. Microscopy examination of the sludge showed a complex mixture of various rod-shaped and coccoid bacterial pluslong filaments and verymobile curved rods. In addition free-living anaerobic ciliates and flagellates were also observed. The trophic group population observed in decreasing order of dominance were hydrolytic and acetogenic at 10(6) and sulfate reducing and methanogenic at 10(5). The rate of methane production in anaerobic granular sludge cultivated in growth medium supplement with formate pressurized with H2:CO2 showed a significant increase in methane yield compared with theseed culture containingthe same substrate and atmosphere of N2:CO2. Similar rates of methane production were observed when the growth medium was supplemented with acetate pressurized either with H2:CO2 or N2:CO2. The number of total anaerobic bacteria at 10(7), fecal coliforms and total coliforms at 10(6), and fecal streptococci at 10(3) is based on colony counts on solid media. The four prevalent species of facultative anaerobic gram-negative bacteria that belong to the family of Enterobacteriaceae were identified as Escherichia coli, Esherichia fergusonii, Klebsiella oxytoca, and Citrobacter freundii. The species Aeromonas hydrophila, Aeromonas veronii, Acinetobacter iwoffi and Stenotrophomonas maltophila were the most frequently isolated glucose fermenting and nonfermenting gram-negative bacilli.

  20. Antimicrobial resistance and susceptibility testing of anaerobic bacteria.

    Science.gov (United States)

    Schuetz, Audrey N

    2014-09-01

    Infections due to anaerobic bacteria can be severe and life-threatening. Susceptibility testing of anaerobes is not frequently performed in laboratories, but such testing is important to direct appropriate therapy. Anaerobic resistance is increasing globally, and resistance trends vary by geographic region. An overview of a variety of susceptibility testing methods for anaerobes is provided, and the advantages and disadvantages of each method are reviewed. Specific clinical situations warranting anaerobic susceptibility testing are discussed.

  1. Nitrogen and phosphorus recovery from anaerobic co-digestion residues of poultry manure and maize silage via struvite precipitation.

    Science.gov (United States)

    Yilmazel, Y Dilsad; Demirer, Goksel N

    2013-08-01

    Anaerobic digestion is commonly used for the stabilization of agricultural and animal wastes. However, owing to the stringent environmental criteria, anaerobic digester effluents need to be further treated to reduce nutrient loads to the receiving water bodies. Struvite precipitation is one of the promising techniques applied for this purpose. Yet, in the majority of cases, struvite precipitation is only applied to the liquid phase of anaerobic digester effluents. This study investigated the recovery of nutrients from both the liquid and the solid phases of the phase-separated effluent of a full-scale biogas plant co-digesting poultry manure and maize silage. Struvite precipitation in the liquid phase led to 72.1% and 95.1% average removal efficiencies of ammonium-nitrogen (NH4-N) and orthophosphate respectively. Changing the external phosphorus source did not make any statistically significant difference in nutrient removal. An acidic phosphorus-dissolution process was applied to the solid phase sample to obtain a phosphorus-enriched solution. More than 90.0% of both NH4-N and PO4-P were recovered from the phosphorus-enriched solution with the amendments of magnesium and phosphorus. In the experiments performed without any addition of external magnesium- and phosphorus-containing chemicals, almost complete (99.6%) PO4-P recovery and partial (14.6%) NH4-N recovery were obtained. The results of this study could contribute to the understanding of nutrient recovery from anaerobic digestion residues of manure and agricultural wastes by struvite precipitation.

  2. Distinct Dual C-Cl Isotope Fractionation Patterns during Anaerobic Biodegradation of 1,2-Dichloroethane: Potential To Characterize Microbial Degradation in the Field.

    Science.gov (United States)

    Palau, J; Yu, R; Hatijah Mortan, S; Shouakar-Stash, O; Rosell, M; Freedman, D L; Sbarbati, C; Fiorenza, S; Aravena, R; Marco-Urrea, E; Elsner, M; Soler, A; Hunkeler, D

    2017-03-07

    This study investigates, for the first time, dual C-Cl isotope fractionation during anaerobic biodegradation of 1,2-dichloroethane (1,2-DCA) via dihaloelimination by Dehalococcoides and Dehalogenimonas-containing enrichment cultures. Isotopic fractionation of 1,2-DCA (εbulk(C) and εbulk(Cl)) for Dehalococcoides (-33.0 ± 0.4‰ and -5.1 ± 0.1‰) and Dehalogenimonas-containing microcosms (-23 ± 2‰ and -12.0 ± 0.8‰) resulted in distinctly different dual element C-Cl isotope correlations (Λ = Δδ(13)C/Δδ(37)Cl ≈ εbulk(C)/εbulk(Cl)), 6.8 ± 0.2 and 1.89 ± 0.02, respectively. Determined isotope effects and detected products suggest that the difference on the obtained Λ values for biodihaloelimination could be associated with a different mode of concerted bond cleavage rather than two different reaction pathways (i.e., stepwise vs concerted). Λ values of 1,2-DCA were, for the first time, determined in two field sites under reducing conditions (2.1 ± 0.1 and 2.2 ± 2.9). They were similar to the one obtained for the Dehalogenimonas-containing microcosms (1.89 ± 0.02) and very different from those reported for aerobic degradation pathways in a previous laboratory study (7.6 ± 0.1 and 0.78 ± 0.03). Thus, this study illustrates the potential of a dual isotope analysis to differentiate between aerobic and anaerobic biodegradation pathways of 1,2-DCA in the field and suggests that this approach might also be used to characterize dihaloelimination of 1,2-DCA by different bacteria, which needs to be confirmed in future studies.

  3. Exocellular electron transfer in anaerobic microbial communities

    NARCIS (Netherlands)

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

    2006-01-01

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

  4. An anaerobic mitochondrion that produces hydrogen

    NARCIS (Netherlands)

    Boxma, Brigitte; Graaf, Rob M. de; Staay, Georg W.M. van der; Alen, Theo A. van; Ricard, Guenola; Gabaldón, Toni; Hoek, Angela H.A.M. van; Moon-van der Staay, Seung Yeo; Koopman, Werner J.H.; Hellemond, Jaap J. van; Tielens, Aloysius G.M.; Friedrich, Thorsten; Veenhuis, Marten; Huynen, Martijn A.; Hackstein, Johannes H.P.

    2005-01-01

    Hydrogenosomes are organelles that produce ATP and hydrogen, and are found in various unrelated eukaryotes, such as anaerobic flagellates, chytridiomycete fungi and ciliates. Although all of these organelles generate hydrogen, the hydrogenosomes from these organisms are structurally and metabolicall

  5. Anaerobic Toxicity of Cationic Silver Nanoparticles

    Data.gov (United States)

    U.S. Environmental Protection Agency — Toxicity data for the impact of nano-silver on anaerobic degradation. This dataset is associated with the following publication: Gitipour, A., S. Thiel, K. Scheckel,...

  6. Anaerobic membrane bioreactor under extreme conditions (poster)

    NARCIS (Netherlands)

    Munoz Sierra, J.D.; De Kreuk, M.K.; Spanjers, H.; Van Lier, J.B.

    2013-01-01

    Membrane bioreactors ensure biomass retention by the application of micro or ultrafiltration processes. This allows operation at high sludge concentrations. Previous studies have shown that anaerobic membrane bioreactors is an efficient way to retain specialist microorganisms for treating wastewater

  7. Anammox enrichment from reject water on blank biofilm carriers and carriers containing nitrifying biomass: operation of two moving bed biofilm reactors (MBBR).

    Science.gov (United States)

    Zekker, Ivar; Rikmann, Ergo; Tenno, Toomas; Lemmiksoo, Vallo; Menert, Anne; Loorits, Liis; Vabamäe, Priit; Tomingas, Martin; Tenno, Taavo

    2012-07-01

    The anammox bacteria were enriched from reject water of anaerobic digestion of municipal wastewater sludge onto moving bed biofilm reactor (MBBR) system carriers-the ones initially containing no biomass (MBBR1) as well as the ones containing nitrifying biomass (MBBR2). Duration of start-up periods of the both reactors was similar (about 100 days), but stable total nitrogen (TN) removal efficiency occurred earlier in the system containing nitrifying biomass. Anammox TN removal efficiency of 70% was achieved by 180 days in both 20 l volume reactors at moderate temperature of 26.0°C. During the steady state phase of operation of MBBRs the average TN removal efficiencies and maximum TN removal rates in MBBR1 were 80% (1,000 g-N/m(3)/day, achieved by 308 days) and in MBBR2 85% (1,100 g-N/m(3)/day, achieved by 266 days). In both reactors mixed bacterial cultures were detected. Uncultured Planctomycetales bacterium clone P4, Candidatus Nitrospira defluvii and uncultured Nitrospira sp. clone 53 were identified by PCR-DGGE from the system initially containing blank biofilm carriers as well as from the nitrifying biofilm system; from the latter in addition to these also uncultured ammonium oxidizing bacterium clone W1 and Nitrospira sp. clone S1-62 were detected. FISH analysis revealed that anammox microorganisms were located in clusters in the biofilm. Using previously grown nitrifying biofilm matrix for anammox enrichment has some benefits over starting up the process from zero, such as less time for enrichment and protection against severe inhibitions in case of high substrate loading rates.

  8. 'Low-acid' sulfide oxidation using nitrate-enriched groundwater

    Science.gov (United States)

    Donn, Michael; Boxall, Naomi; Reid, Nathan; Meakin, Rebecca; Gray, David; Kaksonen, Anna; Robson, Thomas; Shiers, Denis

    2016-04-01

    where pH remains neutral. The "low-acid" oxidation of sulfides with nitrate as an electron acceptor has been demonstrated at the laboratory scale. In 90-day microcosm respirometry experiments, we exposed a mixture of pulverized quartz and pyrite -rich ore to natural, high-nitrate groundwater and inoculated the microcosms with a culture of aerobic and anaerobic nitrate-dependent iron and sulfur-oxidising microorganisms, which were enriched from ore, groundwater and activated waste water. Incubations were performed under both oxic and anoxic conditions, in addition to abiotic controls. Initial results show that oxidation of the sulfides under nitrate-rich and microbially enhanced conditions does produce less acid than the same material under oxic conditions, and to some degree can match the models as long as oxygen ingress can be controlled. These results are the focus of further research into how this process can be enhanced and whether it can be applied in the field. Nitrate-driven oxidation of sulfides could potentially be used as a new approach to reduce acid generation and leaching of contaminants from waste dumps, in a passive or actively managed process designed to deplete and/or ameliorate (i.e. through surface passivation) the mineralogical hazard. Developing our understanding of biological aspects of these processes may also allow testing of longer-term "bio-caps" for various tailings and dump materials.

  9. Methanethiol Removal from Biogas by Biological Conversion in an Anaerobic Biotrickling Filter

    Institute of Scientific and Technical Information of China (English)

    王佳佳; 张卫江; 徐姣

    2015-01-01

    In this study, methanethiol(MT)-degradation bacteria were cultivated by using MT, methanol and trimethylamine as carbon sources under anaerobic conditions. It was found that the batch bacteria used MT and metha-nol as carbon sources grew faster than those used trimethylamine. The enriched bacteria used MT and methanol as the carbon sources were respectively inoculated in different biotrickling filters. The biological conversion performance of MT under anaerobic conditions was investigated in biotrickling filters. The results showed that the performance of the biotrickling filter inoculated with the bacteria enriched using MT was better than that inoculated with the bacteria en-riched using methanol. When the inlet concentration of MT was 0.005vol%(50,ppm), the empty bed residence time was 50 s, pH value was 8.0, and the flow rate of the nutrient solution was 10 L/h, the removal efficiency of MT reached 95.3%. Adding methanol stimulated the growth of the biomass and the degradation of MT, but caused that some bacteria only degrading methanol outcompeted the bacteria only degrading MT. The concentration of sodium bicarbonate in the nutrient solution needed to be controlled lower than 30 g/L, otherwise, it would be harmful to the degradation of MT.

  10. Long-term operation of microbial electrosynthesis cell reducing CO2 to multi-carbon chemicals with a mixed culture avoiding methanogenesis.

    Science.gov (United States)

    Bajracharya, Suman; Yuliasni, Rustiana; Vanbroekhoven, Karolien; Buisman, Cees J N; Strik, David P B T B; Pant, Deepak

    2017-02-01

    In microbial electrosynthesis (MES), CO2 can be reduced preferably to multi-carbon chemicals by a biocathode-based process which uses electrochemically active bacteria as catalysts. A mixed anaerobic consortium from biological origin typically produces methane from CO2 reduction which circumvents production of multi-carbon compounds. This study aimed to develop a stable and robust CO2 reducing biocathode from a mixed culture inoculum avoiding the methane generation. An effective approach was demonstrated based on (i) an enrichment procedure involving inoculum pre-treatment and several culture transfers in H2:CO2 media, (ii) a transfer from heterotrophic to autotrophic growth and (iii) a sequential batch operation. Biomass growth and gradual acclimation to CO2 electro-reduction accomplished a maximum acetate production rate of 400mgLcatholyte(-1)d(-1) at -1V (vs. Ag/AgCl). Methane was never detected in more than 300days of operation. Accumulation of acetate up to 7-10gL(-1) was repeatedly attained by supplying (80:20) CO2:N2 mixture at -0.9 to -1V (vs. Ag/AgCl). In addition, ethanol and butyrate were also produced from CO2 reduction. Thus, a robust CO2 reducing biocathode can be developed from a mixed culture avoiding methane generation by adopting the specific culture enrichment and operation procedures without the direct addition of chemical inhibitor.

  11. EFFECT OF MUSIC ON ANAEROBIC EXERCISE PERFORMANCE

    OpenAIRE

    Atan, T.

    2013-01-01

    For years, mostly the effects of music on cardiorespiratory exercise performance have been studied, but a few studies have examined the effect of music on anaerobic exercise. The purpose of this study was to assess the effect of listening to music and its rhythm on anaerobic exercise: on power output, heart rate and the concentration of blood lactate. 28 male subjects were required to visit the laboratory on 6 occasions, each separated by 48 hours. Firstly, each subject performed the Running-...

  12. Anaerobic digester for treatment of organic waste

    Energy Technology Data Exchange (ETDEWEB)

    Sharma, V. K. [Indian Insitute of Technology, Delhi (India)]|[ENEA, Centro Ricerche Trisaia, Matera (Italy); Fortuna, F.; Canditelli, M.; Cornacchia, G. [ENEA, Centro Ricerche Trisaia, Matera (Italy). Dipt. Ambiente; Farina, R. [ENEA, centro Ricerche ``Ezio Clementel``, Bologna (Italy). Dipt. Ambiente

    1997-09-01

    The essential features of both new and more efficient reactor systems and their appropriate applications for various organic waste management situations, description of several working plants are discussed in the present communication. It is hoped that significant development reported here would be useful in opening a new vista to the application of anaerobic biotechnology for the waste treatment of both low/high organic strength and specialized treatment for toxic substances, using appropriate anaerobic methods.

  13. Anaerobic digestion foaming causes – A review

    OpenAIRE

    Ganidi, Nafsika; Tyrrel, Sean F.; Cartmell, Elise

    2009-01-01

    Anaerobic digestion foaming has been encountered in several sewage treatment plants in the UK. Foaming has raised major concerns for the water companies due to significant impacts on process efficiency and operational costs. Several foaming causes have been identified over the past few years by researchers. However, the supporting experimental information is limited and in some cases absent. The present report aims to provide a detailed review of the current anaerobic digestion foaming proble...

  14. SLEEP DEPRIVATION INDUCED ANXIETY AND ANAEROBIC PERFORMANCE

    Directory of Open Access Journals (Sweden)

    Selma Arzu Vardar

    2007-12-01

    Full Text Available The aim of this study was to investigate the effects of sleep deprivation induced anxiety on anaerobic performance. Thirteen volunteer male physical education students completed the Turkish version of State Anxiety Inventory and performed Wingate anaerobic test for three times: (1 following a full-night of habitual sleep (baseline measurements, (2 following 30 hours of sleep deprivation, and (3 following partial-night sleep deprivation. Baseline measurements were performed the day before total sleep deprivation. Measurements following partial sleep deprivation were made 2 weeks later than total sleep deprivation measurements. State anxiety was measured prior to each Wingate test. The mean state anxiety following total sleep deprivation was higher than the baseline measurement (44.9 ± 12.9 vs. 27.6 ± 4.2, respectively, p = 0.02 whereas anaerobic performance parameters remained unchanged. Neither anaerobic parameters nor state anxiety levels were affected by one night partial sleep deprivation. Our results suggest that 30 hours continuous wakefulness may increase anxiety level without impairing anaerobic performance, whereas one night of partial sleep deprivation was ineffective on both state anxiety and anaerobic performance

  15. Anaerobic reductive dechlorination of tetrachloroethene: how can dual Carbon-Chlorine isotopic measurements help elucidating the underlying reaction mechanism?

    Science.gov (United States)

    Badin, Alice; Buttet, Géraldine; Maillard, Julien; Holliger, Christof; Hunkeler, Daniel

    2014-05-01

    Chlorinated ethenes (CEs) such as tetrachloroethene (PCE) are common persistent groundwater contaminants. Among clean-up strategies applied to sites affected by such pollution, bioremediation has been considered with a growing interest as it represents a cost-effective, environmental friendly approach. This technique however sometimes leads to an incomplete and slow biodegradation of CEs resulting in an accumulation of toxic metabolites. Understanding the reaction mechanisms underlying anaerobic reductive dechlorination would thus help assessing PCE biodegradation in polluted sites. Stable isotope analysis can provide insight into reaction mechanisms. For chlorinated hydrocarbons, carbon (C) and chlorine (Cl) isotope data (δ13C and δ37Cl) tend to show a linear correlation with a slope (m ≡ ɛC/ɛCl) characteristic of the reaction mechanism [1]. This study hence aims at exploring the potential of a dual C-Cl isotope approach in the determination of the reaction mechanisms involved in PCE reductive dechlorination. C and Cl isotope fractionation were investigated during anaerobic PCE dechlorination by two bacterial consortia containing members of the Sulfurospirillum genus. The specificity in these consortia resides in the fact that they each conduct PCE reductive dechlorination catalysed by one different reductive dehalogenase, i.e. PceADCE which yields trichloroethene (TCE) and cis-dichloroethene (cDCE), and PceATCE which yields TCE only. The bulk C isotope enrichment factors were -3.6±0.3 o for PceATCE and -0.7±0.1o for PceADCE. The bulk Cl isotope enrichment factors were -1.3±0.2 o for PceATCE and -0.9±0.1 o for PceADCE. When applying the dual isotope approach, two m values of 2.7±0.1 and 0.7±0.2 were obtained for the reductive dehalogenases PceATCE and PceADCE, respectively. These results suggest that PCE can be degraded according to two different mechanisms. Furthermore, despite their highly similar protein sequences, each reductive dehalogenase seems

  16. Understanding how commensal obligate anaerobic bacteria regulate immune functions in the large intestine.

    Science.gov (United States)

    Maier, Eva; Anderson, Rachel C; Roy, Nicole C

    2014-12-24

    The human gastrointestinal tract is colonised by trillions of commensal bacteria, most of which are obligate anaerobes residing in the large intestine. Appropriate bacterial colonisation is generally known to be critical for human health. In particular, the development and function of the immune system depends on microbial colonisation, and a regulated cross-talk between commensal bacteria, intestinal epithelial cells and immune cells is required to maintain mucosal immune homeostasis. This homeostasis is disturbed in various inflammatory disorders, such as inflammatory bowel diseases. Several in vitro and in vivo studies indicate a role for Faecalibacterium prausnitzii, Bacteroides thetaiotaomicron, Bacteroides fragilis, Akkermansia muciniphila and segmented filamentous bacteria in maintaining intestinal immune homeostasis. These obligate anaerobes are abundant in the healthy intestine but reduced in several inflammatory diseases, suggesting an association with protective effects on human health. However, knowledge of the mechanisms underlying the effects of obligate anaerobic intestinal bacteria remains limited, in part due to the difficulty of co-culturing obligate anaerobes together with oxygen-requiring human epithelial cells. By using novel dual-environment co-culture models, it will be possible to investigate the effects of the unstudied majority of intestinal microorganisms on the human epithelia. This knowledge will provide opportunities for improving human health and reducing the risk of inflammatory diseases.

  17. Highly enriched Betaproteobacteria growing anaerobically with p-xylene and nitrate

    DEFF Research Database (Denmark)

    Rotaru, Amelia-Elena; Probian, Christina; Wilkes, Heinz;

    2010-01-01

    -degrading Betaproteobacteria from the Azoarcus-Thauera clade. Complete oxidation p-xylene to CO(2) coupled to denitrification was suggested by quantitative measurements of substrate consumption. Metabolite analysis identified (4-methylbenzyl)succinate and (4-methylphenyl)itaconate, suggesting addition to fumarate...

  18. Enrichment and terminal differentiation of striated muscle progenitors in vitro

    Energy Technology Data Exchange (ETDEWEB)

    Becher, Ulrich M.; Breitbach, Martin; Sasse, Philipp [Institute of Physiology I, Life and Brain Center, University of Bonn, Bonn (Germany); Garbe, Stephan [Department of Radiology, University of Bonn, Bonn (Germany); Ven, Peter F.M. van der [Institute for Cell Biology, University of Bonn, Bonn (Germany); Fuerst, Dieter O., E-mail: dfuerst@uni-bonn.de [Institute for Cell Biology, University of Bonn, Bonn (Germany); Fleischmann, Bernd K., E-mail: bernd.fleischmann@uni-bonn.de [Institute of Physiology I, Life and Brain Center, University of Bonn, Bonn (Germany)

    2009-10-01

    Enrichment and terminal differentiation of mammalian striated muscle cells is severely hampered by fibroblast overgrowth, de-differentiation and/or lack of functional differentiation. Herein we report a new, reproducible and simple method to enrich and terminally differentiate muscle stem cells and progenitors from mice and humans. We show that a single gamma irradiation of muscle cells induces their massive differentiation into structurally and functionally intact myotubes and cardiomyocytes and that these cells can be kept in culture for many weeks. Similar results are also obtained when treating skeletal muscle-derived stem cells and progenitors with Mitomycin C.

  19. Anaerobic Nitrogen Fixers on Mars

    Science.gov (United States)

    Lewis, B. G.

    2000-07-01

    The conversion of atmospheric nitrogen gas to the protein of living systems is an amazing process of nature. The first step in the process is biological nitrogen fixation, the transformation of N2 to NH3. The phenomenon is crucial for feeding the billions of our species on Earth. On Mars, the same process may allow us to discover how life can adapt to a hostile environment, and render it habitable. Hostile environments also exist on Earth. For example, nothing grows in coal refuse piles due to the oxidation of pyrite and marcasite to sulfuric acid. Yet, when the acidity is neutralized, alfalfa and soybean plants develop root nodules typical of symbiotic nitrogen fixation with Rhizobium species possibly living in the pyritic material. When split open, these nodules exhibited the pinkish color of leghemoglobin, a protein in the nodule protecting the active nitrogen-fixing enzyme nitrogenase against the toxic effects of oxygen. Although we have not yet obtained direct evidence of nitrogenase activity in these nodules (reduction of acetylene to ethylene, for example), these findings suggested the possibility that nitrogen fixation was taking place in this hostile, non-soil material. This immediately raises the possibility that freeliving anaerobic bacteria which fix atmospheric nitrogen on Earth, could do the same on Mars.

  20. Anaerobic Microbial Degradation of Hydrocarbons: From Enzymatic Reactions to the Environment.

    Science.gov (United States)

    Rabus, Ralf; Boll, Matthias; Heider, Johann; Meckenstock, Rainer U; Buckel, Wolfgang; Einsle, Oliver; Ermler, Ulrich; Golding, Bernard T; Gunsalus, Robert P; Kroneck, Peter M H; Krüger, Martin; Lueders, Tillmann; Martins, Berta M; Musat, Florin; Richnow, Hans H; Schink, Bernhard; Seifert, Jana; Szaleniec, Maciej; Treude, Tina; Ullmann, G Matthias; Vogt, Carsten; von Bergen, Martin; Wilkes, Heinz

    2016-01-01

    Hydrocarbons are abundant in anoxic environments and pose biochemical challenges to their anaerobic degradation by microorganisms. Within the framework of the Priority Program 1319, investigations funded by the Deutsche Forschungsgemeinschaft on the anaerobic microbial degradation of hydrocarbons ranged from isolation and enrichment of hitherto unknown hydrocarbon-degrading anaerobic microorganisms, discovery of novel reactions, detailed studies of enzyme mechanisms and structures to process-oriented in situ studies. Selected highlights from this program are collected in this synopsis, with more detailed information provided by theme-focused reviews of the special topic issue on 'Anaerobic biodegradation of hydrocarbons' [this issue, pp. 1-244]. The interdisciplinary character of the program, involving microbiologists, biochemists, organic chemists and environmental scientists, is best exemplified by the studies on alkyl-/arylalkylsuccinate synthases. Here, research topics ranged from in-depth mechanistic studies of archetypical toluene-activating benzylsuccinate synthase, substrate-specific phylogenetic clustering of alkyl-/arylalkylsuccinate synthases (toluene plus xylenes, p-cymene, p-cresol, 2-methylnaphthalene, n-alkanes), stereochemical and co-metabolic insights into n-alkane-activating (methylalkyl)succinate synthases to the discovery of bacterial groups previously unknown to possess alkyl-/arylalkylsuccinate synthases by means of functional gene markers and in situ field studies enabled by state-of-the-art stable isotope probing and fractionation approaches. Other topics are Mo-cofactor-dependent dehydrogenases performing O2-independent hydroxylation of hydrocarbons and alkyl side chains (ethylbenzene, p-cymene, cholesterol, n-hexadecane), degradation of p-alkylated benzoates and toluenes, glycyl radical-bearing 4-hydroxyphenylacetate decarboxylase, novel types of carboxylation reactions (for acetophenone, acetone, and potentially also benzene and

  1. Application of dynamic membranes in anaerobic membranes in anaerobic membrane bioreactor systems

    NARCIS (Netherlands)

    Erşahin, M.E.

    2015-01-01

    Anaerobic membrane bioreactors (AnMBRs) physically ensure biomass retention by the application of a membrane filtration process. With growing application experiences from aerobic membrane bioreactors (MBRs), the combination of membrane and anaerobic processes has received much attention and become m

  2. Importance of pre-enrichment media for isolation of Salmonella spp. from swine and poultry

    DEFF Research Database (Denmark)

    Hoorfar, Jeffrey; Baggesen, Dorte Lau

    1998-01-01

    The performance of two new (1-day) culture methods, Salmonella Enrichment Broth (SEB) and Revive, and an alternative pre-enrichment broth, designated Universal pre-enrichment broth (UB), was compared to the internationally accepted buffered peptone water (BPW). The study was directed towards...... detection of Salmonella in 100 faecal samples from porcine and 100 neck-skin samples from poultry. The sensitivity (number of positive cases per method among all the positive cases) of the conventional pre-enrichment in BPW was found to be 0.77 for swine and 0.66 for poultry samples, while a combination...

  3. The effect of outside conditions on anaerobic ammonia oxidation reaction

    Institute of Scientific and Technical Information of China (English)

    YANG Min; WANG Shu-bo

    2016-01-01

    Organic carbon, inorganic carbon, temperature, pH and ORP are all to have a certain influence on the anaerobic ammonia oxidation reaction. We can draw some conclusions on the optimum conditions of anaerobic ammonia oxidation reaction. The optimum temperature of the anaerobic ammonia oxidation reaction is 30-35℃. And the optimum pH of the anaerobic ammonia reaction is 7.5-8.3. The presence of organic matters can affect the anaerobic ammonia reaction, and different organic matters have different influence on it. The concentration of the inorganic carbon also exist great influence on the reaction. High inorganic carbon concentration also can inhibit anaerobic ammonia oxidation reaction.

  4. Enrichment of light hydrocarbon mixture

    Science.gov (United States)

    Yang; Dali; Devlin, David; Barbero, Robert S.; Carrera, Martin E.; Colling, Craig W.

    2010-08-10

    Light hydrocarbon enrichment is accomplished using a vertically oriented distillation column having a plurality of vertically oriented, nonselective micro/mesoporous hollow fibers. Vapor having, for example, both propylene and propane is sent upward through the distillation column in between the hollow fibers. Vapor exits neat the top of the column and is condensed to form a liquid phase that is directed back downward through the lumen of the hollow fibers. As vapor continues to ascend and liquid continues to countercurrently descend, the liquid at the bottom of the column becomes enriched in a higher boiling point, light hydrocarbon (propane, for example) and the vapor at the top becomes enriched in a lower boiling point light hydrocarbon (propylene, for example). The hollow fiber becomes wetted with liquid during the process.

  5. Effects of microbial inhibitors on anaerobic degradation of DDT

    Energy Technology Data Exchange (ETDEWEB)

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

    2004-09-15

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

  6. Biohydrogen production from rotten orange with immobilized mixed culture: Effect of immobilization media for various composition of substrates

    Energy Technology Data Exchange (ETDEWEB)

    Damayanti, Astrilia, E-mail: liasholehasd@gmail.com [Department of Chemical Engineering, Faculty of Engineering, Semarang State University, E1 Building, 2nd floor, Kampus Sekaran, Gunungpati, Semarang 50229 (Indonesia); Department of Chemical Engineering, Faculty of Engineering, Gadjah Mada University, Jl. Grafika No. 2, Kampus UGM, Yogyakarta 55281 (Indonesia); Sarto,; Syamsiah, Siti; Sediawan, Wahyudi B. [Department of Chemical Engineering, Faculty of Engineering, Gadjah Mada University, Jl. Grafika No. 2, Kampus UGM, Yogyakarta 55281 (Indonesia)

    2015-12-29

    Enriched–immobilized mixed culture was utilized to produce biohydrogen in mesophilic condition under anaerobic condition using rotten orange as substrate. The process was conducted in batch reactors for 100 hours. Microbial cultures from three different sources were subject to a series of enrichment and immobilized in two different types of media, i.e. calcium alginate (CA, 2%) and mixture of alginate and activated carbon (CAC, 1:1). The performance of immobilized culture in each media was tested for biohydrogen production using four different substrate compositions, namely orange meat (OM), orange meat added with peel (OMP), orange meat added with limonene (OML), and mixture of orange meat and peel added with limonene (OMPL). The results show that, with immobilized culture in CA, the variation of substrate composition gave significant effect on the production of biohydrogen. The highest production of biohydrogen was detected for substrate containing only orange meet, i.e. 2.5%, which was about 3-5 times higher than biohydrogen production from other compositions of substrate. The use of immobilized culture in CAC in general has increased the hydrogen production by 2-7 times depending on the composition of substrate, i.e. 5.4%, 4.8%, 5.1%, and 4.4% for OM, OMP, OML, and OMPL, respectively. The addition of activated carbon has eliminated the effect of inhibitory compounds in the substrate. The major soluble metabolites were acetic acid, propionic acid, and butyric acid.

  7. Design Enriches Life,Life Breeds Fashion

    Directory of Open Access Journals (Sweden)

    Yun Gong

    2016-11-01

    Full Text Available As society advances and technology changes,fashion design has developed from the initial few exclusive to public demand, and fashion category also covers all walks of life, extends to every area from a single demand to industrialization. Fashion design development, has become an indispensable part of people's lives. Fashion satisfies public demand, reflecting the public interest towards, and designer correctly grasp of fashion and taste of popular motivated the design of continuous innovation to creating new business value, cultural values of the social value of fashion. A precise definition to fashion is hard, cause too broad as it relates to areas, such as products, clothing, entertainment, advertising, decoration, home, fashion is reach into every aspect of our lives. Fashion derived from life elements, convergence elements of era and innovation into your life all over again, this cycle forward and updated, fully embodies the understanding of art and life. Design enriches life, life gave birth to fashion.

  8. Effect of Different Filling Materials in Anammox Bacteria Enrichment

    Directory of Open Access Journals (Sweden)

    Dilek ÖZGÜN

    2012-12-01

    Full Text Available Purpose: Anaerobic ammonium oxidation (Anammox is a process that ammonium as electron donor is oxidized to nitrogen gas using nitrite as electron acceptor. Compared to conventional nitrification-denitrification processes, this process is used less oxygen and no organic material (methanol, glucose. However, the slow growth rate of Anammox bacteria (11-30 days is disadvantages. Therefore, batch reactors have been carried out in these bacteria enrichment. In this study continuously operated upflow anaerobic sludge reactor (UASB using different filling materials disposing of sensitive and slow-growing Anammox bacteria out of the system is purposed. Design and Methods: System is operated up-flow column reactor at 2 days hydraulic retention time (HRT in 45 days. In this study, ceramic stones and Linpor filling material are used. Using synthetic wastewater containing ammonium and nitrite, Ar/CO2 anaerobic conditions (95/5% supplied with gas. System is operated at a temperature 253 C in UASB. Temperature, pH, ammonia-nitrogen and nitrite nitrogen are measured. Results: Both filling material reactors are operated in 45 days. Ceramic stones filling reactor is observed quickly reaches 90% were used reactor ammonium removal. The ammonium nitrogen removal was slower in Linpor filling materials reactor. Nitrite removal is reached up to 90% in both the reactor. When compared to the stoichiometric equation in Linpor was composed of large amounts of nitrate. At the end of 25 days the results were similar to ceramic stone filling reactor with Linpor filling material reactors. Conclusions and Original Value: Anammox process as from nitrogen removal processes was discovered in 1995. Anammox bacteria that make up this process due to very low growth rates of microbial bacteria in the system must be kept in the system. Most of the studies in the literature, these bacteria enrichment stage is started instead of a continuous batch reactor system. In this study

  9. 31 CFR 540.316 - Uranium enrichment.

    Science.gov (United States)

    2010-07-01

    ... 31 Money and Finance: Treasury 3 2010-07-01 2010-07-01 false Uranium enrichment. 540.316 Section... FOREIGN ASSETS CONTROL, DEPARTMENT OF THE TREASURY HIGHLY ENRICHED URANIUM (HEU) AGREEMENT ASSETS CONTROL REGULATIONS General Definitions § 540.316 Uranium enrichment. The term uranium enrichment means the process...

  10. N2O production by ammonia oxidizing bacteria in an enriched nitrifying sludge linearly depends on inorganic carbon concentration.

    Science.gov (United States)

    Peng, Lai; Ni, Bing-Jie; Ye, Liu; Yuan, Zhiguo

    2015-05-01

    The effect of inorganic carbon (IC) on nitrous oxide (N2O) production by ammonia oxidizing bacteria (AOB) was investigated over a concentration range of 0-12 mmol C/L, encompassing typical IC levels in a wastewater treatment reactors. The AOB culture was enriched along with nitrite-oxidizing bacteria (NOB) in a sequencing batch reactor (SBR) to perform complete nitrification. Batch experiments were conducted with continuous carbon dioxide (CO2) stripping or at controlled IC concentrations. The results revealed a linear relationship between N2O production rate (N2OR) and IC concentration (R(2) = 0.97) within the IC range studied, suggesting a substantial effect of IC on N2O production by AOB. Similar results were also obtained with an AOB culture treating anaerobic sludge digestion liquor. The fundamental mechanism responsible for this dependency is unclear; however, in agreement with previous studies, it was observed that the ammonia oxidation rate (AOR) was also influenced by the IC concentration, which could be well described by the Monod kinetics. These resulted in an exponential relationship between N2OR and AOR, as previously observed in experiments where AOR was altered by varying dissolved oxygen and ammonia concentrations. It is therefore possible that IC indirectly affected N2OR by causing a change in AOR. The observation in this study indicates that alkalinity (mostly contributed by IC) could be a significant factor influencing N2O production and should be taken into consideration in estimating and mitigating N2O emissions in wastewater treatment systems.

  11. ANAEROBIC BIOLOGICAL TREATMENT OF PRODUCED WATER

    Energy Technology Data Exchange (ETDEWEB)

    John R. Gallagher

    2001-07-31

    During the production of oil and gas, large amounts of water are brought to the surface and must be disposed of in an environmentally sensitive manner. This is an especially difficult problem in offshore production facilities where space is a major constraint. The chief regulatory criterion for produced water is oil and grease. Most facilities have little trouble meeting this criterion using conventional oil-water separation technologies. However, some operations have significant amounts of naphthenic acids in the water that behave as oil and grease but are not well removed by conventional technologies. Aerobic biological treatment of naphthenic acids in simulated-produced water has been demonstrated by others; however, the system was easily overloaded by the large amounts of low-molecular-weight organic acids often found in produced waters. The objective of this research was to determine the ability of an anaerobic biological system to treat these organic acids in a simulated produced water and to examine the potential for biodegradation of the naphthenic acids in the anaerobic environment. A small fixed-film anaerobic biological reactor was constructed and adapted to treat a simulated produced water. The bioreactor was tubular, with a low-density porous glass packing material. The inocula to the reactor was sediment from a produced-water holding pond from a municipal anaerobic digester and two salt-loving methanogenic bacteria. During start-up, the feed to the reactor contained glucose as well as typical produced-water components. When glucose was used, rapid gas production was observed. However, when glucose was eliminated and the major organic component was acetate, little gas was generated. Methane production from acetate may have been inhibited by the high salt concentrations, by sulfide, or because of the lack, despite seeding, of microbes capable of converting acetate to methane. Toluene, a minor component of the produced water (0.1 g/L) was removed in the

  12. Genome sequence of Victivallis vadensis ATCC BAA-548, an anaerobic bacterium from the phylum Lentisphaerae, isolated from the human gastro-intestinal tract

    Energy Technology Data Exchange (ETDEWEB)

    Van Passel, Mark W.J. [Wageningen University and Research Centre, The Netherlands; Kant, Ravi [University of Helsinki; Palva, Airi [University of Helsinki; Lucas, Susan [U.S. Department of Energy, Joint Genome Institute; Copeland, A [U.S. Department of Energy, Joint Genome Institute; Lapidus, Alla L. [U.S. Department of Energy, Joint Genome Institute; Glavina Del Rio, Tijana [U.S. Department of Energy, Joint Genome Institute; Dalin, Eileen [U.S. Department of Energy, Joint Genome Institute; Tice, Hope [U.S. Department of Energy, Joint Genome Institute; Bruce, David [Los Alamos National Laboratory (LANL); Goodwin, Lynne A. [Los Alamos National Laboratory (LANL); Pitluck, Sam [U.S. Department of Energy, Joint Genome Institute; Davenport, Karen W. [Los Alamos National Laboratory (LANL); Sims, David [Los Alamos National Laboratory (LANL); Detter, J. Chris [U.S. Department of Energy, Joint Genome Institute; Han, Cliff [Los Alamos National Laboratory (LANL); Larimer, Frank W [ORNL; Land, Miriam L [ORNL; Hauser, Loren John [ORNL; Kyrpides, Nikos C [U.S. Department of Energy, Joint Genome Institute; Ovchinnikova, Galina [U.S. Department of Energy, Joint Genome Institute; Richardson, Paul [U.S. Department of Energy, Joint Genome Institute; De Vos, Willem M. [Wageningen University and Research Centre, The Netherlands; Smidt, Hauke [Wageningen University and Research Centre, The Netherlands; Zoetendal, Erwin G. [Wageningen University and Research Centre, The Netherlands

    2011-01-01

    Victivallis vadensis ATCC BAA-548 represents the first cultured representative from the novel phylum Lentisphaerae, a deep-branching bacterial lineage. Few cultured bacteria from this phylum are known, and V. vadensis therefore represents an important organism for evolutionary studies. V. vadensis is a strictly anaerobic sugar-fermenting isolate from the human gastro-intestinal tract.

  13. Oxygen Effects in Anaerobic Digestion - II

    Directory of Open Access Journals (Sweden)

    Deshai Botheju

    2010-04-01

    Full Text Available Standard models describing bio-gasification using anaerobic digestion do not include necessary processes to describe digester dynamics under the conditions of oxygen presence. Limited oxygenation in anaerobic digestion can sometimes be beneficial. The oxygen effects included anaerobic digestion model, ADM 1-Ox, was simulated against experimental data obtained from laboratory scale anaerobic digesters operated under different oxygenation conditions. ADM 1-Ox predictions are generally in good agreement with the trends of the experimental data. ADM 1-Ox simulations suggest the existence of an optimum oxygenation level corresponding to a peak methane yield. The positive impact of oxygenation on methane yield is more pronounced at conditions characterized by low hydrolysis rate coefficients (slowly degradable feed and low biomass concentrations. The optimum oxygenation point moves towards zero when the hydrolysis rate coefficient and the biomass concentration increase. Accordingly, the impact of oxygenation on methane yield can either be positive or negative depending on the digestion system characteristics. The developed ADM 1-Ox model can therefore be a valuable tool for recognizing suitable operating conditions for achieving the maximum benefits from partial aeration in anaerobic digestion.

  14. Experimental otitis media in gerbils and chinchillas with Streptococcus pneumoniae, Haemophilus influenzae, and other aerobic and anaerobic bacteria.

    OpenAIRE

    Fulghum, R S; Brinn, J E; Smith, A M; Daniel, H J; Loesche, P J

    1982-01-01

    To ascertain the usefulness of Mongolian gerbils as an inbred model for otitis media, 52 Mongolian gerbils (Meriones unguiculatus, strain MONT/Tum) were compared with 26 chinchillas (Chinchilla laniger) for susceptibility to Streptococcus pneumoniae type 3. Haemophilus influenzae type b, and a polymicrobic culture including anaerobes (Streptococcus intermedius, Propionibacterium acnes, Staphylococcus epidermidis, and Corynebacterium sp.). Organisms were inoculated percutaneously into the supe...

  15. Anaerobic denitrification in fungi from the coastal marine sediments off Goa, India

    Digital Repository Service at National Institute of Oceanography (India)

    Manohar, C.S.; Raghukumar, C.

    . Res.: 113(1); 2009; 100-109 Anaerobic denitrification in fungi from the coastal marine sediments off Goa, India Cathrine Sumathi J*, Chandralata Raghukumar National Institute of Oceanography, (Council of Scientific and Industrial Research) Dona... grateful to Dr. Shoun H. for lending us the culture # MT-811 for our studies. We are thankful to Dr Seshagiri Raghukumar for his critical review of the manuscript and for helping us in the identification of the fungi. This is NIO contribution number...

  16. Building a Culture of Achievement

    Science.gov (United States)

    Major, Marc R.

    2009-01-01

    Culture is the social and intergenerational glue that defines, connects, sustains, and enriches the members of successful communities--including schools and classrooms. A classroom culture is a psychological atmosphere that nurtures and shapes students' attitudes about their own identity, classes, school, and learning in general. Classroom culture…

  17. Dissimilatory reduction of perchlorate and other common pollutants by a consortium enriched from tidal flats of the Yellow Sea

    Institute of Scientific and Technical Information of China (English)

    Nirmala Bardiya; Jae-Ho Bae

    2015-01-01

    Objective: To enrich a facultative anaerobic bacterial consortium from the Yellow Sea and assess its ability to reduce perchlorate and other co-pollutants. Methods: Bacterial consortium collected from the tidal flats of the Yellow Sea was enriched in an anoxic medium containing perchlorate as the electron (e-) acceptor and acetate as the electron (e-) donor. The enriched consortium was then tested for perchlorate reduction under different perchlorate concentrations and in the presence of nitrate by using standard anaerobic techniques. The complete enzymatic reduction of perchlorate to chloride was confirmed by chlorite dismutation. Ability of the consortium to grow with alternate e- acceptors was also tested with acetate as the e- donor. Results: The enriched consortium could rapidly reduce perchlorate up to the initial concentration of 25.65 mmol/L. In the presence of nitrate, perchlorate reduction did not occur immediately and reduction of nitrate started after a lag phase, with concomitant accumulation of nitrite. The perchlorate-enriched consortium could reduce chlorate, oxygen, Cr (VI), and selenate as the alternate e- acceptors but failed to utilize sulfate, thiosulfate, sulfite, and nitrite. Conclusions: The consortium from the tidal flats of the Yellow Sea could reduce perchlorate and co-contaminants such as chlorate, nitrate, Cr (VI), and selenate under heterotrophic conditions with acetate as the e- donor and carbon source. While perchlorate was completely dismutated into innocuous chloride and oxygen, accumulation of nitrite occurred during the reduction of nitrate.

  18. Nonproteinogenic D-amino acids at millimolar concentrations are a toxin for anaerobic microorganisms relevant to early Earth and other anoxic planets.

    Science.gov (United States)

    Nixon, Sophie L; Cockell, Charles S

    2015-03-01

    The delivery of extraterrestrial organics to early Earth provided a potentially important source of carbon and energy for microbial life. Optically active organic compounds of extraterrestrial origin exist in racemic form, yet life on Earth has almost exclusively selected for L- over D-enantiomers of amino acids. Although D-enantiomers of proteinogenic amino acids are known to inhibit aerobic microorganisms, the role of concentrated nonproteinogenic meteoritic D-amino acids on anaerobic metabolisms relevant to early Earth and other anoxic planets such as Mars is unknown. Here, we test the inhibitory effect of D-enantiomers of two nonproteinogenic amino acids common to carbonaceous chondrites, norvaline and α-aminobutyric acid, on microbial iron reduction. Three pure strains (Geobacter bemidjiensis, Geobacter metallireducens, Geopsychrobacter electrodiphilus) and an iron-reducing enrichment culture were grown in the presence of 10 mM D-enantiomers of both amino acids. Further tests were conducted to assess the inhibitory effect of these D-amino acids at 1 and 0.1 mM. The presence of 10 mM D-norvaline and D-α-aminobutyric acid inhibited microbial iron reduction by all pure strains and the enrichment. G. bemidjiensis was not inhibited by either amino acid at 0.1 mM, but D-α-aminobutyric acid still inhibited at 1 mM. Calculations using published meteorite accumulation rates to the martian surface indicate D-α-aminobutyric acid may have reached inhibitory concentrations in little over 1000 years during peak infall. These data show that, on a young anoxic planet, the use of one enantiomer over another may render the nonbiological enantiomer an environmental toxin. Processes that generate racemic amino acids in the environment, such as meteoritic infall or impact synthesis, would have been toxic processes and could have been a selection pressure for the evolution of early racemases.

  19. Biological hydrogen production by moderately thermophilic anaerobic bacteria

    Energy Technology Data Exchange (ETDEWEB)

    HP Goorissen; AJM Stams [Laboratory of Microbiology, Wageningen University and Research centre Wageningen (Netherlands)

    2006-07-01

    This study focuses on the biological production of hydrogen at moderate temperatures (65-75 C) by anaerobic bacteria. A survey was made to select the best (moderate) thermophiles for hydrogen production from cellulolytic biomass. From this survey we selected Caldicellulosiruptor saccharolyticus (a gram-positive bacterium) and Thermotoga elfii (a gram-negative bacterium) as potential candidates for biological hydrogen production on mixtures of C{sub 5}-C{sub 6} sugars. Xylose and glucose were used as model substrates to describe growth and hydrogen production from hydrolyzed biomass. Mixed substrate utilization in batch cultures revealed differences in the sequence of substrate consumption and in catabolites repression of the two microorganisms. The regulatory mechanisms of catabolites repression in these microorganisms are not known yet. (authors)

  20. [Prolonged cultivation of an anaerobic bacterial community producing hydrogen].

    Science.gov (United States)

    Belokopytov, B F; Ryzhmanova, Ia V; Laurinavichius, K S; Shcherbakova, V A

    2012-01-01

    This paper studies various methods of long-term maintenance of the process of hydrogen evolution during the growth of an aerobic bacterial community on a starch-containing environment. When cultured in separable trip fermentation mode for 72 days, from 0.10 to 0.23 H2/l of medium/day was formed. The regime of regular reseeding lasted more than 100 days, forming an average of 0.81 1 H2/l of medium/day. The advantages and disadvantages of different methods of microbial hydrogen production during a dark starch fermentation process are presented. From the obtained H2 forming microbial communities, we isolated an anaerobic spore-forming bacterium (strain BF). Phylogenetic analysis of the 16S RNA gene sequence of the new strain showed that according to its genotype it belongs to the Clostridium butyricum species.

  1. Anaerobic lipid degradation through acidification and methanization.

    Science.gov (United States)

    Kim, Ijung; Kim, Sang-Hyoun; Shin, Hang-Sik; Jung, Jin-Young

    2010-01-01

    In biological wastewater treatment high lipid concentration is known to inhibit microorganisms and cause active biomass flotation. To reduce lipid inhibition, a two-phase anaerobic system, consisting of an anaerobic sequencing batch reactor (ASBR) and an upflow anaerobic sludge blanket (UASB) reactor, was applied to synthetic dairy wastewater. During 153 days of operation, the two-phase system showed stable performance in lipid degradation. In the ASBR, a 13% lipid removal efficiency and 10% double bond removal efficiency were maintained. In the UASB, the chemical oxygen demand (COD), lipid and volatile fatty acid (VFA) removal efficiencies were more than 80%, 70% and 95%, respectively, up to organic loading rate 6.5 g COD/L/day. There were no operational problems such as serious scum formation or sludge washout. Protein degradation occurred prior to degradation during acidogenesis.

  2. Biochemistry and physiology of anaerobic bacteria

    Energy Technology Data Exchange (ETDEWEB)

    NONE

    2000-05-18

    We welcome you to The Power of Anaerobes. This conference serves two purposes. One is to celebrate the life of Harry D. Peck, Jr.,who was born May 18, 1927 and would have celebrated his 73rd birthday at this conference. He died November 20, 1998. The second is to gather investigators to exchange views within the realm of anaerobic microbiology, an area in which tremendous progress has been seen during recent years. It is sufficient to mention discoveries of a new form of life (the archaea), hyper or extreme thermophiles, thermophilic alkaliphiles and anaerobic fungi. With these discoveries has come a new realization about physiological and metabolic properties of microorganisms, and this in turn has demonstrated their importance for the development, maintenance and sustenance of life on Earth.

  3. Kinetics and modeling of anaerobic digestion process

    DEFF Research Database (Denmark)

    2003-01-01

    Anaerobic digestion modeling started in the early 1970s when the need for design and efficient operation of anaerobic systems became evident. At that time not only was the knowledge about the complex process of anaerobic digestion inadequate but also there were computational limitations. Thus......, the first models were very simple and consisted of a limited number of equations. During the past thirty years much research has been conducted on the peculiarities of the process and on the factors that influence it on the one hand while an enormous progress took place in computer science on the other....... The combination of both parameters resulted in the development of more and more concise and complex models. In this chapter the most important models found in the literature are described starting from the simplest and oldest to the more recent and complex ones....

  4. Multivariate monitoring of anaerobic co-digestion

    DEFF Research Database (Denmark)

    Madsen, Michael; Holm-Nielsen, Jens Bo

    Anaerobic digestion processes for production of renewable energy in the form of biogas, and in the future hydrogen, are becoming increasingly important worldwide. Sustainable solutions for renewable energy production systems are given high political priority, amongst other things due to global...... warming and environmental concerns. Anaerobic digestion applied in agriculture can simultaneously convert heterogeneous biomasses and wastes from the primary agricultural sector and from the bio processing industries, for instance food processing, pharma, and biofuel production, into valuable organic...... fertiliser and renewable energy. Meanwhile, in order for the biogas sector to become a significant player in the energy supply chain, the anaerobic digestion process has to be controlled to a greater extent than what is implemented as state-of-the-art today. Through application of the philosophy behind...

  5. The effect of tannic compounds on anaerobic wastewater treatment.

    NARCIS (Netherlands)

    Field, J.A.

    1989-01-01

    Anaerobic wastewater treatment is an alternative to the conventional aerobic treatment processes for the removal of easily biodegradable organic matter in medium to high strength industrial wastestreams. Anaerobic treatment has several advantages, however one important disadvantage is the high sensi

  6. Operation of three parallel AN/AO processes to enrich denitrifying phosphorus removing bacteria for low strength wastewater treatment

    Institute of Scientific and Technical Information of China (English)

    XIA Si-qing; LIU Hong-bo

    2006-01-01

    Three parallel anaerobic-anoxic/anaerobic-aerobic (AN/AO) processes were developed to enrich denitrifying phosphorus removal bacteria (DPB) for low strength wastewater treatment. The main body of the parallel AN/AO process consists of an AN(anaerobic-anoxic) process and an AO (anaerobic-aerobic) process. In the AO process, the common phosphorus accumulating organisms (PAOs) was dominate, while in the AN process, DPB was dominate. The volume of anaerobic zone(Vana):anoxic zone(Vano):aerobic zone (Vaer) for the parallel AN/AO process is 1:1:1 in contrast with a Vana:Vaer and Vano,:Vaer of 1:2 and 1:4 for a traditional biological nutrient removal process (BNR). Process 3 excels in the 3 processes on the basis of COD, TN and TP removal. For 4 month operation, the effluent COD concentration of process 3 did not exceed 60 mg/L; the effluent TN concentration of process 3 was lower than 15 mg/L; and the effluent TP concentration of process 3 was lower than 1 mg/L.

  7. Robust regulation of anaerobic digestion processes.

    Science.gov (United States)

    Mailleret, L; Bernard, O; Steyer, J P

    2003-01-01

    This paper deals with the problem of controlling anaerobic digestion processes. A two-step (i.e. acidogenesis-methanization) mass balance model is considered for a 1 m3 fixed bed digester treating industrial wine distillery wastewater. The control law aims at regulating the organic pollution level while avoiding washout of biomass. To this end, a simple output feedback controller is considered which regulates a variable strongly related to the Chemical Oxygen Demand (COD). Numerical simulations assuming noisy measurements first illustrate the robustness of this control procedure. Then, the regulating procedure is implemented on the considered anaerobic digestion process in order to validate and demonstrate its efficiency in real life experiments.

  8. Anaerobic bacteria, the colon and colitis.

    Science.gov (United States)

    Roediger, W E

    1980-02-01

    Anaerobic bacteria constitute more than 90% of the bacteria in the colon. An anaerobic environment is needed to maintain their growth and the production of short-chain fatty acids by these bacteria from carbohydrates. Short-chain fatty acids are rapidly absorbed and essential for metabolic as well as functional welfare of the colonic mucosa. The importance of these acids in water absorption and in the patogenesis of colitis is discussed in relation to the concept of "energy deficiency diseases" of the colonic mucosa.

  9. The Pasteur effect in facultative anaerobic metazoa.

    Science.gov (United States)

    Schmidt, H; Kamp, G

    1996-05-15

    The existence and the regulatory mechanisms of the Pasteur effect in facultative anaerobic metazoa are discussed. There are three reasons for the controversy surrounding this phenomenon. 1) The different definitions of the Pasteur effect, 2) the antagonistic effect of metabolic depression and its species specific response to hypoxia, as well as 3) the laboratory-specific differences in the experimental procedures for analyzing the Pasteur effect and its regulation. This review aims to clarify the confusion about the existence of the Pasteur effect in facultative anaerobic metazoa and to offer possible molecular mechanisms.

  10. Life cycle assessment of energy from waste via anaerobic digestion: a UK case study.

    Science.gov (United States)

    Evangelisti, Sara; Lettieri, Paola; Borello, Domenico; Clift, Roland

    2014-01-01

    Particularly in the UK, there is potential for use of large-scale anaerobic digestion (AD) plants to treat food waste, possibly along with other organic wastes, to produce biogas. This paper presents the results of a life cycle assessment to compare the environmental impacts of AD with energy and organic fertiliser production against two alternative approaches: incineration with energy production by CHP and landfill with electricity production. In particular the paper investigates the dependency of the results on some specific assumptions and key process parameters. The input Life Cycle Inventory data are specific to the Greater London area, UK. Anaerobic digestion emerges as the best treatment option in terms of total CO2 and total SO2 saved, when energy and organic fertiliser substitute non-renewable electricity, heat and inorganic fertiliser. For photochemical ozone and nutrient enrichment potentials, AD is the second option while incineration is shown to be the most environmentally friendly solution. The robustness of the model is investigated with a sensitivity analysis. The most critical assumption concerns the quantity and quality of the energy substituted by the biogas production. Two key issues affect the development and deployment of future anaerobic digestion plants: maximising the electricity produced by the CHP unit fuelled by biogas and to defining the future energy scenario in which the plant will be embedded.

  11. Microbial community structure and dynamics during anaerobic digestion of various agricultural waste materials.

    Science.gov (United States)

    Ziganshin, Ayrat M; Liebetrau, Jan; Pröter, Jürgen; Kleinsteuber, Sabine

    2013-06-01

    The influence of the feedstock type on the microbial communities involved in anaerobic digestion was investigated in laboratory-scale biogas reactors fed with different agricultural waste materials. Community composition and dynamics over 2 months of reactors' operation were investigated by amplicon sequencing and profiling terminal restriction fragment length polymorphisms of 16S rRNA genes. Major bacterial taxa belonged to the Clostridia and Bacteroidetes, whereas the archaeal community was dominated by methanogenic archaea of the orders Methanomicrobiales and Methanosarcinales. Correlation analysis revealed that the community composition was mainly influenced by the feedstock type with the exception of a temperature shift from 38 to 55 °C which caused the most pronounced community shifts. Bacterial communities involved in the anaerobic digestion of conventional substrates such as maize silage combined with cattle manure were relatively stable and similar to each other. In contrast, special waste materials such as chicken manure or Jatropha press cake were digested by very distinct and less diverse communities, indicating partial ammonia inhibition or the influence of other inhibiting factors. Anaerobic digestion of chicken manure relied on syntrophic acetate oxidation as the dominant acetate-consuming process due to the inhibition of aceticlastic methanogenesis. Jatropha as substrate led to the enrichment of fiber-degrading specialists belonging to the genera Actinomyces and Fibrobacter.

  12. Anaerobic oxidation of methane by sulfate in hypersaline groundwater of the Dead Sea aquifer.

    Science.gov (United States)

    Avrahamov, N; Antler, G; Yechieli, Y; Gavrieli, I; Joye, S B; Saxton, M; Turchyn, A V; Sivan, O

    2014-11-01

    Geochemical and microbial evidence points to anaerobic oxidation of methane (AOM) likely coupled with bacterial sulfate reduction in the hypersaline groundwater of the Dead Sea (DS) alluvial aquifer. Groundwater was sampled from nine boreholes drilled along the Arugot alluvial fan next to the DS. The groundwater samples were highly saline (up to 6300 mm chlorine), anoxic, and contained methane. A mass balance calculation demonstrates that the very low δ(13) CDIC in this groundwater is due to anaerobic methane oxidation. Sulfate depletion coincident with isotope enrichment of sulfur and oxygen isotopes in the sulfate suggests that sulfate reduction is associated with this AOM. DNA extraction and 16S amplicon sequencing were used to explore the microbial community present and were found to be microbial composition indicative of bacterial sulfate reducers associated with anaerobic methanotrophic archaea (ANME) driving AOM. The net sulfate reduction seems to be primarily controlled by the salinity and the available methane and is substantially lower as salinity increases (2.5 mm sulfate removal at 3000 mm chlorine but only 0.5 mm sulfate removal at 6300 mm chlorine). Low overall sulfur isotope fractionation observed ((34) ε = 17 ± 3.5‰) hints at high rates of sulfate reduction, as has been previously suggested for sulfate reduction coupled with methane oxidation. The new results demonstrate the presence of sulfate-driven AOM in terrestrial hypersaline systems and expand our understanding of how microbial life is sustained under the challenging conditions of an extremely hypersaline environment.

  13. Enhanced phosphorus recovery and biofilm microbial community changes in an alternating anaerobic/aerobic biofilter.

    Science.gov (United States)

    Tian, Qing; Ong, Say Kee; Xie, Xuehui; Li, Fang; Zhu, Yanbin; Wang, Feng Rui; Yang, Bo

    2016-02-01

    The operation of an alternating anaerobic/aerobic biofilter (AABF), treating synthetic wastewater, was modified to enhance recovery of phosphorus (P). The AABF was periodically fed with an additional carbon source during the anaerobic phase to force the release of biofilm-sequestered P which was then harvested and recovered. A maximum of 48% of the total influent P was found to be released in the solution for recovery. Upon implementation of periodic P bio-sequestering and P harvesting, the predominant bacterial communities changed from β-Proteobacteria to γ-Proteobacteria groups. The genus Pseudomonas of γ-Proteobacteria was found to enrich greatly with 98% dominance. Dense intracellular poly-P granules were found within the cells of the biofilm, confirming the presence of P accumulating organisms (PAOs). Periodic addition of a carbon source to the AABF coupled with intracellular P reduction during the anaerobic phase most probably exerted environmental stress in the selection of Pseudomonas PAOs over PAOs of other phylogenic types. Results of the study provided operational information on the selection of certain microbial communities for P removal and recovery. This information can be used to further advance P recovery in biofilm systems such as the AABFs.

  14. The Influence of Hydration on Anaerobic Performance: A Review

    Science.gov (United States)

    Kraft, Justin A.; Green, James M.; Bishop, Phillip A.; Richardson, Mark T.; Neggers, Yasmin H.; Leeper, James D.

    2012-01-01

    This review examines the influence of dehydration on muscular strength and endurance and on single and repeated anaerobic sprint bouts. Describing hydration effects on anaerobic performance is difficult because various exercise modes are dominated by anaerobic energy pathways, but still contain inherent physiological differences. The critical…

  15. Stability of anaerobic reactors under micro-aeration conditions

    Energy Technology Data Exchange (ETDEWEB)

    Fernandez-Polanco, M.; Perez, S.; Diaz, I.; Fernandez-Polanco, F.

    2009-07-01

    Oxidation of sulphide in anaerobic bioreactors by introducing limited amounts of oxygen provides a relatively simple strategy for reducing the levels of sulphite in anaerobic digesters (biogas and effluent). The introduction of limited amounts of air is a general practice in agricultural anaerobic digesters, it is estimated that worldwide over 3.000 units are operated under such conditions. (Author)

  16. Anaerobic Digestion. Student Manual. Biological Treatment Process Control.

    Science.gov (United States)

    Carnegie, John W., Ed.

    This student manual contains the textual material for a four-lesson unit on anaerobic digestion control. Areas addressed include: (1) anaerobic sludge digestion (considering the nature of raw sludge, purposes of anaerobic digestion, the results of digestion, types of equipment, and other topics); (2) digester process control (considering feeding…

  17. Microalgae Cultivation on Anaerobic Digestate of Municipal Wastewater, Sewage Sludge and Agro-Waste

    Directory of Open Access Journals (Sweden)

    Luca Zuliani

    2016-10-01

    Full Text Available Microalgae are fast-growing photosynthetic organisms which have the potential to be exploited as an alternative source of liquid fuels to meet growing global energy demand. The cultivation of microalgae, however, still needs to be improved in order to reduce the cost of the biomass produced. Among the major costs encountered for algal cultivation are the costs for nutrients such as CO2, nitrogen and phosphorous. In this work, therefore, different microalgal strains were cultivated using as nutrient sources three different anaerobic digestates deriving from municipal wastewater, sewage sludge or agro-waste treatment plants. In particular, anaerobic digestates deriving from agro-waste or sewage sludge treatment induced a more than 300% increase in lipid production per volume in Chlorella vulgaris cultures grown in a closed photobioreactor, and a strong increase in carotenoid accumulation in different microalgae species. Conversely, a digestate originating from a pilot scale anaerobic upflow sludge blanket (UASB was used to increase biomass production when added to an artificial nutrient-supplemented medium. The results herein demonstrate the possibility of improving biomass accumulation or lipid production using different anaerobic digestates.

  18. Antarctic strict anaerobic microbiota from Deschampsia antarctica vascular plants rhizosphere reveals high ecology and biotechnology relevance.

    Science.gov (United States)

    Peixoto, Rafael José Marques; Miranda, Karla Rodrigues; Lobo, Leandro Araujo; Granato, Alessandra; de Carvalho Maalouf, Pedro; de Jesus, Hugo Emiliano; Rachid, Caio T C C; Moraes, Saulo Roni; Dos Santos, Henrique Fragoso; Peixoto, Raquel Silva; Rosado, Alexandre Soares; Domingues, Regina Maria Cavalcanti Pilotto

    2016-11-01

    The Antarctic soil microbial community has a crucial role in the growth and stabilization of higher organisms, such as vascular plants. Analysis of the soil microbiota composition in that extreme environmental condition is crucial to understand the ecological importance and biotechnological potential. We evaluated the efficiency of isolation and abundance of strict anaerobes in the vascular plant Deschampsia antarctica rhizosphere collected in the Antarctic's Admiralty Bay and associated biodiversity to metabolic perspective and enzymatic activity. Using anaerobic cultivation methods, we identified and isolated a range of microbial taxa whose abundance was associated with Plant Growth-Promoting Bacteria (PGPB) and presences were exclusively endemic to the Antarctic continent. Firmicutes was the most abundant phylum (73 %), with the genus Clostridium found as the most isolated taxa. Here, we describe two soil treatments (oxygen gradient and heat shock) and 27 physicochemical culture conditions were able to increase the diversity of anaerobic bacteria isolates. Heat shock treatment allowed to isolate a high percentage of new species (63.63 %), as well as isolation of species with high enzymatic activity (80.77 %), which would have potential industry application. Our findings contribute to the understanding of the role of anaerobic microbes regarding ecology, evolutionary, and biotechnological features essential to the Antarctic ecosystem.

  19. Study of the role of anaerobic metabolism in succinate production by Enterobacter aerogenes.

    Science.gov (United States)

    Tajima, Yoshinori; Kaida, Kenichi; Hayakawa, Atsushi; Fukui, Keita; Nishio, Yousuke; Hashiguchi, Kenichi; Fudou, Ryosuke; Matsui, Kazuhiko; Usuda, Yoshihiro; Sode, Koji

    2014-09-01

    Succinate is a core biochemical building block; optimizing succinate production from biomass by microbial fermentation is a focus of basic and applied biotechnology research. Lowering pH in anaerobic succinate fermentation culture is a cost-effective and environmentally friendly approach to reducing the use of sub-raw materials such as alkali, which are needed for neutralization. To evaluate the potential of bacteria-based succinate fermentation under weak acidic (pH Enterobacter aerogenes AJ110637, which rapidly assimilates glucose at pH 5.0. Based on the profile of anaerobic products, we constructed single-gene knockout mutants to eliminate the main anaerobic metabolic pathways involved in NADH re-oxidation. These single-gene knockout studies showed that the ethanol synthesis pathway serves as the dominant NADH re-oxidation pathway in this organism. To generate a metabolically engineered strain for succinate production, we eliminated ethanol formation and introduced a heterogeneous carboxylation enzyme, yielding E. aerogenes strain ΔadhE/PCK. The strain produced succinate from glucose with a 60.5% yield (grams of succinate produced per gram of glucose consumed) at pH <6.2 and anaerobic conditions. Thus, we showed the potential of bacteria-based succinate fermentation under weak acidic conditions.

  20. Enrichment of Fusobacteria in Sea Surface Oil Slicks from the Deepwater Horizon Oil Spill.

    Science.gov (United States)

    Gutierrez, Tony; Berry, David; Teske, Andreas; Aitken, Michael D

    2016-07-27

    The Deepwater Horizon (DWH) oil spill led to rapid microbial community shifts in the Gulf of Mexico, including the formation of unprecedented quantities of marine oil snow (MOS) and of a massive subsurface oil plume. The major taxa that bloomed in sea surface oil slicks during the spill included Cycloclasticus, and to a lesser extent Halomonas, Alteromonas, and Pseudoalteromonas-organisms that grow and degrade oil hydrocarbons aerobically. Here, we show that sea surface oil slicks at DWH contained obligate and facultative anaerobic taxa, including members of the obligate anaerobic phylum Fusobacteria that are commonly found in marine sediment environments. Pyrosequencing analysis revealed that Fusobacteria were strongly selected for when sea surface oil slicks were allowed to develop anaerobically. These organisms have been found in oil-contaminated sediments in the Gulf of Mexico, in deep marine oil reservoirs, and other oil-contaminated sites, suggesting they have putative hydrocarbon-degrading qualities. The occurrence and strong selection for Fusobacteria in a lab-based incubation of a sea surface oil slick sample collected during the spill suggests that these organisms may have become enriched in anaerobic zones of suspended particulates, such as MOS. Whilst the formation and rapid sinking of MOS is recognised as an important mechanism by which a proportion of the Macondo oil had been transported to the sea floor, its role in potentially transporting microorganisms, including oil-degraders, from the upper reaches of the water column to the seafloor should be considered. The presence of Fusobacteria on the sea surface-a highly oxygenated environment-is intriguing, and may be explained by the vertical upsurge of oil that provided a carrier to transport these organisms from anaerobic/micro-aerophilic zones in the oil plume or seabed to the upper reaches of the water column. We also propose that the formation of rapidly-sinking MOS may have re-transported these

  1. Environmental impacts of anaerobic digestion and the use of anaerobic residues as soil amendment

    Energy Technology Data Exchange (ETDEWEB)

    Mosey, F.E. [VFA Services Ltd., Herts (United Kingdom)

    1996-01-01

    This paper defines the environmental role of anaerobic digestion within the overall objective of recovering energy from renewable biomass resources. Examples and opportunities for incorporating anaerobic digestion into biomass-to-energy schemes are discussed, together with environmental aspects of anaerobic digestion plants. These include visual, public amenity, pathogens and public health, odor control, and gaseous emissions. Digestate disposal and the benefits of restrictions on recycling organic wastes and biomass residues back to the land are discussed, particularly as they relate to American and European codes of practice and environmental legislation. The paper concludes that anaerobic digestion, if performed in purpose-designed reactors that efficiently recover and use biogas, is an environmentally benign process that can enhance energy recovery and aid the beneficial land use of plant residues in many biomass-to-energy schemes.

  2. Assessment of active methanogenic archaea in a methanol-fed upflow anaerobic sludge blanket reactor.

    Science.gov (United States)

    Cerrillo, Míriam; Morey, Lluís; Viñas, Marc; Bonmatí, August

    2016-12-01

    Methanogenic archaea enrichment of a granular sludge was undertaken in an upflow anaerobic sludge blanket (UASB) reactor fed with methanol in order to enrich methylotrophic and hydrogenotrophic methanogenic populations. A microbial community assessment, in terms of microbial composition and activity-throughout the different stages of the feeding process with methanol and acetate-was performed using specific methanogenic activity (SMA) assays, quantitative real-time polymerase chain reaction (qPCR), and high-throughput sequencing of 16S ribosomal RNA (rRNA) genes from DNA and complementary DNA (cDNA). Distinct methanogenic enrichment was revealed by qPCR of mcrA gene in the methanol-fed community, being two orders of magnitude higher with respect to the initial inoculum, achieving a final mcrA/16S rRNA ratio of 0.25. High-throughput sequencing analysis revealed that the resulting methanogenic population was mainly composed by methylotrophic archaea (Methanomethylovorans and Methanolobus genus), being also highly active according to the RNA-based assessment. SMA confirmed that the methylotrophic pathway, with a direct conversion of methanol to CH4, was the main step of methanol degradation in the UASB. The biomass from the UASB, enriched in methanogenic archaea, may bear great potential as additional inoculum for bioreactors to carry out biogas production and other related processes.

  3. Growth kinetics and competition between Methanosarcina and Methanosaeta in mesophilic anaerobic digestion.

    Science.gov (United States)

    Conklin, Anne; Stensel, H David; Ferguson, John

    2006-05-01

    Methanosarcina species with a high maximum specific growth rate (mumax) and high half-saturation coefficient (KS) and Methanosaeta species with a low mumax and low KS are the only known aceticlastic methanogens. Because of Methanosaeta's low KS, the low acetate concentrations in conventional, mesophilic anaerobic digestion yield Methanosaeta dominance. However, Methanosarcina absorbs increases in acetate more efficiently and thus promotes more stable digestion. This paper tests the hypothesis that decreasing digester feeding frequencies can increase Methanosarcina predominance. Two acetate-fed reactors were established at a 17-day solids retention time. One reactor was fed hourly, and one was fed once daily. Microscopic and molecular methods were used to verify that the hourly fed reactor enriched for Methanosaeta, while the daily fed reactor enriched for Methanosarcina. Growth and substrate-use kinetics were measured for each reactor. A digester overload condition was simulated, and the Methanosarcina-enriched reactor was found to perform better than the Methanosaeta-enriched reactor. These findings indicate that Methanosarcina dominance can be achieved with infrequent feedings, leading to more stable digestion.

  4. Tissue culture: the unrealized potential

    OpenAIRE

    Sato, Gordon

    2007-01-01

    Lack of differentiated functions of the tissue of origin in tissue culture thought to be due to dedifferentiation was shown to be due to selective overgrowth of fibroblasts. Enrichment culture techniques, (alternate animal and culture passage), designed to give the functionally differentiated cells selective advantage over the fibroblasts resulted in a large number of functionally differentiated clonal strains. Thus the dogma of dedifferentiation was destroyed. It is proposed to substitute th...

  5. Biomedical advances from tissue culture

    OpenAIRE

    Okamoto, Tetsuji; Sato, J. Denry; Barnes, David W.; Sato, Gordon H.

    2013-01-01

    The demonstration that the “dedifferentiation” of cells commonly observed in the early days of tissue