WorldWideScience

Sample records for denitrifying enrichment cultures

  1. Anaerobic ferrous oxidation by heterotrophic denitrifying enriched culture.

    Wang, Ru; Zheng, Ping; Xing, Ya-Juan; Zhang, Meng; Ghulam, Abbas; Zhao, Zhi-Qing; Li, Wei; Wang, Lan

    2014-05-01

    Heterotrophic denitrifying enriched culture (DEC) from a lab-scale high-rate denitrifying reactor was discovered to perform nitrate-dependent anaerobic ferrous oxidation (NAFO). The DEC was systematically investigated to reveal their denitrification activity, their NAFO activity, and the predominant microbial population. The DEC was capable of heterotrophic denitrification with methanol as the electron donor, and autotrophic denitrification with ferrous salt as the electron donor named NAFO. The conversion ratios of ferrous-Fe and nitrate-N were 87.41 and 98.74 %, and the consumption Fe/N ratio was 2.3:1 (mol/mol). The maximum reaction velocity and half saturation constant of Fe were 412.54 mg/(l h) and 8,276.44 mg/l, and the counterparts of N were 20.87 mg/(l h) and 322.58 mg/l, respectively. The predominant bacteria were Hyphomicrobium, Thauera, and Flavobacterium, and the predominant archaea were Methanomethylovorans, Methanohalophilus, and Methanolobus. The discovery of NAFO by heterotrophic DEC is significant for the development of wastewater treatment and the biogeochemical iron cycle and nitrogen cycle. PMID:24619339

  2. Degradation of toluene and m-xylene and transformation of o-xylene by denitrifying enrichment cultures.

    Evans, P J; Mang, D T; Young, L Y

    1991-01-01

    Seven different sources of inocula that included sediments, contaminated soils, groundwater, process effluent, and sludge were used to establish enrichment cultures of denitrifying bacteria on benzene, toluene, and xylenes in the absence of molecular oxygen. All of the enrichment cultures demonstrated complete depletion of toluene and partial depletion of o-xylene within 3 months of incubation. The depletion of o-xylene was correlated to and dependent on the metabolism of toluene. No losses of benzene, p-xylene, or m-xylene were observed in these initial enrichment cultures. However, m-xylene was degraded by a subculture that was incubated on m-xylene alone. Complete carbon, nitrogen, and electron balances were determined for the degradation of toluene and m-xylene. These balances showed that these compounds were mineralized with greater than 50% conversion to CO2 and significant assimilation into biomass. Additionally, the oxidation of these compounds was shown to be dependent on nitrate reduction and denitrification. These microbial degradative capabilities appear to be widespread, since the widely varied inoculum sources all yielded similar results. PMID:2014990

  3. Anaerobic oxidation of cholesterol by a denitrifying enrichment.

    Barrandeguy, E; Tarlera, S

    2001-01-01

    Sterols (e.g. cholesterol) present in wool scouring effluent represent the most recalcitrant fraction in anaerobic treatment. This study was conducted to examine the feasibility of removal of this organic load through a denitrifying post-treatment stage. A stable cholesterol-denitrifying enrichment (CHOL-1) was obtained from sludge of a bench-scale upflow sludge bed (USB) denitrifying reactor integrated to a carbon and nitrogen removal system for sanitary landfill leachate. According to the amounts of cholesterol degraded and of nitrite and nitrogen gas formed, the capacity for complete cholesterol oxidation under anaerobic conditions by CHOL-1 can be assumed. Nitrite accumulation observed at a low C/N ratio outlines the importance of determining the optimal C/N ratio for adequate denitrifying reactor performance. The enrichment was partly identified with molecular analysis of cloned 16S rDNA sequences revealing the presence of two groups of bacteria belonging to the beta subclass of the Proteobacteria. According to analysis of sequences, it can be inferred that a yet uncultivated new bacterium is the one responsible for cholesterol oxidation. Results of this study suggest that sludge from a denitrifying reactor treating leachate is potentially useful in a combined anaerobic-anoxic system for degradation of cholesterol that remains after methanogenic treatment. PMID:11575077

  4. Metagenomic analysis of denitrifying wastewater enrichment cultures able to transform the explosive, 3-nitro-1,2,4-triazol-5-one (NTO).

    Eberly, Jed O; Indest, Karl J; Hancock, Dawn E; Jung, Carina M; Crocker, Fiona H

    2016-06-01

    Removal of 3-nitro-1,2,4-triazol-5-one (NTO) was investigated in conjunction with heterotrophic and autotrophic denitrifying growth conditions by a microbial consortium from a wastewater treatment plant. Microcosms were supplemented with molasses, methanol, or thiosulfate. Cultures were passaged twice by transferring 10 % of the culture volume to fresh media on days 11 and 21. Rates of NTO removal were 18.71 ± 0.65, 9.04 ± 2.61, and 4.34 ± 2.72 mg/L/day while rates of nitrate removal were 20.08 ± 1.13, 21.58 ± 1.20, and 24.84 ± 1.26 mg/L/day, respectively, for molasses, methanol, or thiosulfate. Metagenomic analysis showed that Proteobacteria and Firmicutes were the major phyla in the microbial communities. In molasses supplemented cultures, the community profile at the family level changed over time with Pseudomonadaceae the most abundant (67.4 %) at day 11, Clostridiaceae (65.7 %) at day 21, and Sporolactobacillaceae (35.4 %) and Clostridiaceae (41.0 %) at day 29. Pseudomonadaceae was the dominant family in methanol and thiosulfate supplemented cultures from day 21 to 29 with 76.6 and 81.6 % relative abundance, respectively. PMID:27033535

  5. Enrichment of DNRA bacteria in a continuous culture.

    van den Berg, Eveline M; van Dongen, Udo; Abbas, Ben; van Loosdrecht, Mark Cm

    2015-10-01

    Denitrification and dissimilatory nitrate reduction to ammonium (DNRA) are competing microbial nitrate-reduction processes. The occurrence of DNRA has been shown to be effected qualitatively by various parameters in the environment. A more quantitative understanding can be obtained using enrichment cultures in a laboratory reactor, yet no successful DNRA enrichment culture has been described. We showed that a stable DNRA-dominated enrichment culture can be obtained in a chemostat system. The enrichment was based on the hypothesis that nitrate limitation is the dominant factor in selecting for DNRA. First, a conventional denitrifying culture was enriched from activated sludge, with acetate and nitrate as substrates. Next, the acetate concentration in the medium was increased to obtain nitrate-limiting conditions. As a result, conversions shifted from denitrification to DNRA. In this selection of a DNRA culture, two important factors were the nitrate limitation and a relatively low dilution rate (0.026 h(-1)). The culture was a highly enriched population of Deltaproteobacteria most closely related to Geobacter lovleyi, based on 16S rRNA gene sequencing (97% similarity). We established a stable and reproducible cultivation method for the enrichment of DNRA bacteria in a continuously operated reactor system. This enrichment method allows to further investigate the DNRA process and address the factors for competition between DNRA and denitrification, or other N-conversion pathways. PMID:25909972

  6. Distribution of baroduric, psychrotrophic and culturable nitrifying and denitrifying bacteria in the Central Indian Basin

    LokaBharathi, P.A.; PradeepRam, A.S.; Nair, S.; Nath, B.N.; Chandramohan, D.

    The abundance of baroduric, culturable nitrifying and denitrifying bacteria in the deep-sea cores of Central Indian Basin (CIB) at ca 5000 m depth was investigated. Analysis of 8 cores, sampled between 10 degrees 00 minutes S and 75 degrees 55...

  7. Enrichment of denitrifying methanotrophic bacteria for application after direct low-temperature anaerobic sewage treatment

    Kampman, Christel, E-mail: christel.kampman@wur.nl [Sub-department of Environmental Technology, Wageningen University, P.O. Box 17, 6700 AA, Wageningen (Netherlands); Hendrickx, Tim L.G. [Sub-department of Environmental Technology, Wageningen University, P.O. Box 17, 6700 AA, Wageningen (Netherlands); Luesken, Francisca A.; Alen, Theo A. van; Op den Camp, Huub J.M.; Jetten, Mike S.M. [Department of Microbiology, Institute for Water and Wetland Research, Radboud University Nijmegen, Heyendaalseweg 135, 6525 AJ Nijmegen (Netherlands); Zeeman, Grietje; Buisman, Cees J.N.; Temmink, Hardy [Sub-department of Environmental Technology, Wageningen University, P.O. Box 17, 6700 AA, Wageningen (Netherlands)

    2012-08-15

    Highlights: Black-Right-Pointing-Pointer A new concept for low-temperature anaerobic sewage treatment is proposed. Black-Right-Pointing-Pointer In this concept, denitrification and methane oxidation are performed by Methylomirabilis oxyfera. Black-Right-Pointing-Pointer The bacteria were enriched from fresh water sediment using sequencing fed-batch reactors. Black-Right-Pointing-Pointer The volumetric consumption rate has to be increased by an order of magnitude for practical application. Black-Right-Pointing-Pointer Further research should focus on systems with improved biomass retention. - Abstract: Despite many advantages of anaerobic sewage treatment over conventional activated sludge treatment, it has not yet been applied in temperate zones. This is especially because effluent from low-temperature anaerobic treatment contains nitrogen and dissolved methane. The presence of nitrogen and methane offers the opportunity to develop a reactor in which methane is used as electron donor for denitrification. Such a reactor could be used in a new concept for low-temperature anaerobic sewage treatment, consisting of a UASB-digester system, a reactor for denitrification coupled to anaerobic methane oxidation, and a nitritation reactor. In the present study denitrifying methanotrophic bacteria similar to 'Candidatus Methylomirabilis oxyfera' were enriched. Maximum volumetric nitrite consumption rates were 33.5 mg NO{sub 2}{sup -}-N/L d (using synthetic medium) and 37.8 mg NO{sub 2}{sup -}-N/L d (using medium containing effluent from a sewage treatment plant), which are similar to the maximum rate reported so far. Though the goal was to increase the rates, in both reactors, after reaching these maximum rates, volumetric nitrite consumption rates decreased in time. Results indicate biomass washout may have significantly decelerated enrichment. Therefore, to obtain higher volumetric consumption rates, further research should focus on systems with complete biomass retention.

  8. Enrichment of denitrifying methanotrophic bacteria for application after direct low-temperature anaerobic sewage treatment

    Highlights: ► A new concept for low-temperature anaerobic sewage treatment is proposed. ► In this concept, denitrification and methane oxidation are performed by Methylomirabilis oxyfera. ► The bacteria were enriched from fresh water sediment using sequencing fed-batch reactors. ► The volumetric consumption rate has to be increased by an order of magnitude for practical application. ► Further research should focus on systems with improved biomass retention. - Abstract: Despite many advantages of anaerobic sewage treatment over conventional activated sludge treatment, it has not yet been applied in temperate zones. This is especially because effluent from low-temperature anaerobic treatment contains nitrogen and dissolved methane. The presence of nitrogen and methane offers the opportunity to develop a reactor in which methane is used as electron donor for denitrification. Such a reactor could be used in a new concept for low-temperature anaerobic sewage treatment, consisting of a UASB-digester system, a reactor for denitrification coupled to anaerobic methane oxidation, and a nitritation reactor. In the present study denitrifying methanotrophic bacteria similar to ‘Candidatus Methylomirabilis oxyfera’ were enriched. Maximum volumetric nitrite consumption rates were 33.5 mg NO2−-N/L d (using synthetic medium) and 37.8 mg NO2−-N/L d (using medium containing effluent from a sewage treatment plant), which are similar to the maximum rate reported so far. Though the goal was to increase the rates, in both reactors, after reaching these maximum rates, volumetric nitrite consumption rates decreased in time. Results indicate biomass washout may have significantly decelerated enrichment. Therefore, to obtain higher volumetric consumption rates, further research should focus on systems with complete biomass retention.

  9. Anaerobic Toluene Activation by Benzylsuccinate Synthase in a Highly Enriched Methanogenic Culture

    Beller, Harry R.; Edwards, Elizabeth A

    2000-01-01

    Permeabilized cells of a highly enriched, toluene-mineralizing, methanogenic culture catalyzed the addition of toluene to fumarate to form benzylsuccinate under anaerobic conditions. The specific in vitro rate of benzylsuccinate formation was >85% of the specific in vivo rate of toluene consumption. This is the first report of benzylsuccinate synthase activity in a methanogenic culture; the activity has previously been reported to occur in denitrifying, sulfate-reducing, and anoxygenic photot...

  10. Enrichment of denitrifying methanotrophic bacteria from Taihu sediments by a membrane biofilm bioreactor at ambient temperature.

    Wang, Shenghui; Wu, Qing; Lei, Ting; Liang, Peng; Huang, Xia

    2016-03-01

    Denitrification coupled to anaerobic methane oxidation is a recently discovered process performed by bacteria affiliated to the NC10 phylum. These microorganisms could play important roles in the energy-efficient way of anaerobic wastewater treatment where residual dissolved methane might be removed at the expense of nitrate or nitrite. The difficulty to enrich these microorganisms due to a slow growth rate, especially at low temperatures, limited its application in engineering field. In this study, an NC10 bacteria community was enriched from Taihu sediments by a membrane biofilm bioreactor at ambient temperature of 10-25 °C. After 13 months enrichment, the maximum denitrification rate of the enriched culture reached 0.54 mM day(-1) for nitrate and 1.06 mM day(-1) for nitrite. Anaerobic methane oxidation coupled denitrification was estimated from the (13)C-labeled CO2 ((13)CO2) production during batch incubations with (13)CH4. Furthermore, analysis of 16S rRNA genes clone library confirmed the presence of NC10 phylum bacteria and fluorescence in situ hybridization showed that NC10 bacteria dominated the reactor. All of the results indicated the NC10 bacteria community was competitive in terms of treating nitrate-contaminated water or wastewater under natural conditions. PMID:26578374

  11. Mineralization of Trichloroethylene by Heterotrophic Enrichment Cultures

    Fliermans, C. B.; Phelps, T J; Ringelberg, D.; Mikell, A. T.; White, D C

    1988-01-01

    Microbial consortia capable of aerobically degrading more than 99% of exogenous trichloroethylene (TCE) (50 mg/liter) were collected from TCE-contaminated subsurface sediments and grown in enrichment cultures. TCE at concentrations greater than 300 mg/liter was not degraded, nor was TCE used by the consortia as a sole energy source. Energy sources which permitted growth included tryptone-yeast extract, methanol, methane, and propane. The optimum temperature range for growth and subsequent TCE...

  12. 21 CFR 866.2330 - Enriched culture medium.

    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. Bacteriophages May Bias Outcome of Bacterial Enrichment Cultures

    Muniesa, Maite; Blanch, Anicet R; Lucena, Francisco; Jofre, Juan de

    2005-01-01

    Enrichment cultures are widely used for the isolation of bacteria in clinical, biotechnological, and environmental studies. However, competition, relative growth rates, or inhibitory effects may alter the outcome of enrichment cultures, causing the phenomenon known as enrichment bias. Bacteriophages are a major component in many microbial systems, and it abounds in natural settings. This abundance means that bacteriophages are likely to be present in many laboratory enrichment cultures. Our h...

  14. Highly enriched Betaproteobacteria growing anaerobically with p-xylene and nitrate

    Rotaru, Amelia-Elena; Probian, Christina; Wilkes, Heinz; Harder, Jens

    2010-01-01

    The identity of the microorganisms capable of anaerobic p-xylene degradation under denitrifying conditions is hitherto unknown. Here, we report highly enriched cultures of freshwater denitrifying bacteria that grow anaerobically with p-xylene as the sole organic carbon source and electron donor....... Long curved rods, with 95% 16S rRNA gene sequence identity to Denitratisoma oestradiolicum, dominated the enrichment cultures (>91% of all cells), as detected by phylotype-specific probes. These Rhodocyclaceae microorganisms were distantly related to other denitrifying hydrocarbon-degrading...

  15. Enrichment of high ammonia tolerant methanogenic culture

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

    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...... population was derived from a full scale biogas reactor (Hashøj, Denmark), fed with 75% animal manure and 25% food industries organic waste. Basal anaerobic medium was used for the enrichment along with sodium acetate (1 g HAc L-1) as a carbon source. Fluorescence insitu hybridization (FISH) was used to...

  16. Enrichment and Short Term Culture of the Ovine Gonocyte

    Rongfeng Li

    2011-01-01

    Full Text Available The aim of this study was to investigate the effects of two types of ovine testis cells population as feeder cell on in vitro culture of the enriched ovine gonocytes. The feeder cell populations were prepared from 5-6 months old ovine testis. The 1-2 months old neonatal rams were used to isolate germ cells through a two step enzymatic digestion followed by differential plating for SSCs enrichment. Isolated and enriched cells were characterized by using PLZF and VASA antibody. During the 1st week of culture, gonocyte formed pairs and chains of type A spermatogonia. After 1 week, colonies started to increase in size. About 2 weeks later, more colonies in type II feeder group kept undifferentiated and looks more effective regarding colony formation of spermatogonia compared with type I feeder group in the ovine spermatogonial stem cell culture system.

  17. Protein Chips for Detection of Salmonella spp. from Enrichment Culture.

    Poltronieri, Palmiro; Cimaglia, Fabio; De Lorenzis, Enrico; Chiesa, Maurizio; Mezzolla, Valeria; Reca, Ida Barbara

    2016-01-01

    Food pathogens are the cause of foodborne epidemics, therefore there is a need to detect the pathogens in food productions rapidly. A pre-enrichment culture followed by selective agar plating are standard detection methods. Molecular methods such as qPCR have provided a first rapid protocol for detection of pathogens within 24 h of enrichment culture. Biosensors also may provide a rapid tool to individuate a source of Salmonella contamination at early times of pre-enrichment culture. Forty mL of Salmonella spp. enrichment culture were processed by immunoseparation using the Pathatrix, as in AFNOR validated qPCR protocols. The Salmonella biosensor combined with immunoseparation showed a limit of detection of 100 bacteria/40 mL, with a 400 fold increase to previous results. qPCR analysis requires processing of bead-bound bacteria with lysis buffer and DNA clean up, with a limit of detection of 2 cfu/50 μL. Finally, a protein chip was developed and tested in screening and identification of 5 common pathogen species, Salmonella spp., E. coli, S. aureus, Campylobacter spp. and Listeria spp. The protein chip, with high specificity in species identification, is proposed to be integrated into a Lab-on-Chip system, for rapid and reproducible screening of Salmonella spp. and other pathogen species contaminating food productions. PMID:27110786

  18. Protein Chips for Detection of Salmonella spp. from Enrichment Culture

    Palmiro Poltronieri

    2016-04-01

    Full Text Available Food pathogens are the cause of foodborne epidemics, therefore there is a need to detect the pathogens in food productions rapidly. A pre-enrichment culture followed by selective agar plating are standard detection methods. Molecular methods such as qPCR have provided a first rapid protocol for detection of pathogens within 24 h of enrichment culture. Biosensors also may provide a rapid tool to individuate a source of Salmonella contamination at early times of pre-enrichment culture. Forty mL of Salmonella spp. enrichment culture were processed by immunoseparation using the Pathatrix, as in AFNOR validated qPCR protocols. The Salmonella biosensor combined with immunoseparation showed a limit of detection of 100 bacteria/40 mL, with a 400 fold increase to previous results. qPCR analysis requires processing of bead-bound bacteria with lysis buffer and DNA clean up, with a limit of detection of 2 cfu/50 μL. Finally, a protein chip was developed and tested in screening and identification of 5 common pathogen species, Salmonella spp., E. coli, S. aureus, Campylobacter spp. and Listeria spp. The protein chip, with high specificity in species identification, is proposed to be integrated into a Lab-on-Chip system, for rapid and reproducible screening of Salmonella spp. and other pathogen species contaminating food productions.

  19. Chloroform degradation in methanogenic methanol enrichment cultures and by Methanosarcina barkeri 227.

    Bagley, D M; Gossett, J M

    1995-01-01

    The effects of methanol addition and consumption on chloroform degradation rate and product distribution in methanogenic methanol enrichment cultures and in cultures of Methanosarcina barkeri 227 were investigated. Degradation of chloroform with initial concentrations up to 27.3 microM in enrichment cultures and 4.8 microM in pure cultures was stimulated by the addition of methanol. However, methanol consumption was inhibited by as little as 2.5 microM chloroform in enrichment cultures and 0....

  20. Immunological identification and distribution of dissimilatory heme cd1 and nonheme copper nitrite reductases in denitrifying bacteria.

    Coyne, M S; Arunakumari, A; Averill, B.A.; Tiedje, J. M.

    1989-01-01

    Polyclonal antibodies were used to identify heme or copper nitrite reductases in the following groups: 23 taxonomically diverse denitrifiers from culture collections, 100 numerically dominant denitrifiers from geographically diverse environments, and 51 denitrifiers from a culture collection not selected for denitrification. Antisera were raised against heme nitrite reductases from Pseudomonas aeruginosa and Pseudomonas stutzeri and against copper nitrite reductase from Achromobacter cyclocla...

  1. Anaerobic oxidation of acetylene by estuarine sediments and enrichment cultures

    Culbertson, Charles W.; Zehnder, Alexander J. B.; Oremland, Ronald S.

    1981-01-01

    Acetylene disappeared from the gas phase of anaerobically incubated estuarine sediment slurries, and loss was accompanied by increased levels of carbon dioxide. Acetylene loss was inhibited by chloramphenicol, air, and autoclaving. Addition of14C2H2to slurries resulted in the formation of14CO2and the transient appearance of14C-soluble intermediates, of which acetate was a major component. Acetylene oxidation stimulated sulfate reduction; however, sulfate reduction was not required for the loss of C2H2to occur. Enrichment cultures were obtained which grew anaerobically at the expense of C2H2.

  2. Anaerobic oxidation of acetylene by estuarine sediments and enrichment cultures

    Acetylene disappeared from the gas phase of anaerobically incubated estuarine sediment slurries, and loss was accompanied by increased levels of carbon dioxide. Acetylene loss was inhibited by chloroamphenicol, air, and autoclaving. Addition of 14C2H2 to slurries resulted in the formation of 14CO2 and the transient appearance of 14C-soluble intermediates, of which acetate was a major component. Acetylene oxidation stimulated sulfate reduction; however, sulfate reduction was not required for the loss of C2H2 to occur. Enrichment cultures were obtained which grew anaerobically at the expense of C2H2

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

    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. PMID:25280176

  4. Effect of Enrichment Medium on Real-time Detection of Salmonella enterica from Lettuce and Tomato Enrichment Cultures

    vThree commonly used enrichment broths for detection of Salmonella (Buffered Peptone Water BPW, Tryptic Soy Broth TSB, and Universal Preenrichment Broth UPB) were compared for use in real time SYBR Green PCR detection of Salmonella introduced into enrichment cultures made from store bought let...

  5. Bordetella avium antibiotic resistance, novel enrichment culture, and antigenic characterization.

    Beach, Nathan M; Thompson, Seth; Mutnick, Rachel; Brown, Lisa; Kettig, Gina; Puffenbarger, Robyn; Stockwell, Stephanie B; Miyamoto, David; Temple, Louise

    2012-11-01

    Bordetella avium continues to be an economic issue in the turkey industry as the causative agent of bordetellosis, which often leads to serious secondary infections. This study presents a broad characterization of the antibiotic resistance patterns in this diverse collection of B. avium strains collected over the past thirty years. In addition, the plasmid basis for the antibiotic resistance was characterized. The antibiotic resistance pattern allowed the development of a novel enrichment culture method that was subsequently employed to gather new isolates from diseased turkeys and a healthy sawhet owl. While a healthy turkey flock was shown to seroconvert by four weeks-of-age, attempts to culture B. avium from healthy turkey poults were unsuccessful. Western blot of B. avium strains using pooled serum from diseased and healthy commercial turkey flocks revealed both antigenic similarities and differences between strains. In sum, the work documents the continued exposure of commercial turkey flocks to B. avium and the need for development of an effective, inexpensive vaccine to control spread of the disease. PMID:22721730

  6. Heterotrophic nitrification among denitrifiers.

    Castignetti, D; Hollocher, T C

    1984-01-01

    Twelve denitrifying bacteria representing six genera were tested for an ability to nitrify pyruvic oxime heterotrophically. Six of these bacteria exhibited appreciable nitrification activity, yielding as much as 5.8 mM nitrite and little or no nitrate when grown in a mineral salts medium containing 7 mM pyruvic oxime and 0.05% yeast extract. Of the six active bacteria, four (Pseudomonas denitrificans, Pseudomonas aeruginosa, and two strains of Pseudomonas fluorescens) could grow on yeast extr...

  7. Selective Enrichment Media Bias the Types of Salmonella enterica Strains Isolated from Mixed Strain Cultures and Complex Enrichment Broths

    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-o...

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

    Luo, Gang; Angelidaki, Irini

    2012-01-01

    existing natural gas grid. The current study presents a new biological method for biogas upgrading in a separate biogas reactor, containing enriched hydrogenotrophic methanogens and fed with biogas and hydrogen. Both mesophilic- and thermophilic anaerobic cultures were enriched to convert CO2 to CH4 by...... addition of H2. Enrichment at thermophilic temperature (55°C) resulted in CO2 and H2 bioconversion rate of 320 mL CH4/(gVSS h), which was more than 60% higher than that under mesophilic temperature (37°C). Different dominant species were found at mesophilic- and thermophilic-enriched cultures, as revealed...

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

    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. PMID:26846292

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

    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.

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

    Alessandra Valle, Daur; Francisco, Klimak Jr.; Laura Lcia, Cogo; Gislene Digenes, Boto; Cristina Leise Bastos, Monteiro; Libera Maria, Dalla Costa.

    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.

  12. Enhanced production and isotope enrichment of recombinant glycoproteins produced in cultured mammalian cells

    Skelton, David; Goodyear, Abbey [Florida State University, Department of Chemistry and Biochemistry (United States); Ni, DaQun; Walton, Wendy J.; Rolle, Myron; Hare, Joan T. [Florida State University, Institute of Molecular Biophysics (United States); Logan, Timothy M., E-mail: tlogan@fsu.ed [Florida State University, Department of Chemistry and Biochemistry (United States)

    2010-10-15

    NMR studies of post-translationally modified proteins are complicated by the lack of an efficient method to produce isotope enriched recombinant proteins in cultured mammalian cells. We show that reducing the glucose concentration and substituting glutamate for glutamine in serum-free medium increased cell viability while simultaneously increasing recombinant protein yield and the enrichment of non-essential amino acids compared to culture in unmodified, serum-free medium. Adding dichloroacetate, a pyruvate dehydrogenase kinase inhibitor, further improves cell viability, recombinant protein yield, and isotope enrichment. We demonstrate the method by producing partially enriched recombinant Thy1 glycoprotein from Lec1 Chinese hamster ovary (CHO) cells using U-{sup 13}C-glucose and {sup 15}N-glutamate as labeled precursors. This study suggests that uniformly {sup 15}N,{sup 13}C-labeled recombinant proteins may be produced in cultured mammalian cells starting from a mixture of labeled essential amino acids, glucose, and glutamate.

  13. Enhanced production and isotope enrichment of recombinant glycoproteins produced in cultured mammalian cells

    NMR studies of post-translationally modified proteins are complicated by the lack of an efficient method to produce isotope enriched recombinant proteins in cultured mammalian cells. We show that reducing the glucose concentration and substituting glutamate for glutamine in serum-free medium increased cell viability while simultaneously increasing recombinant protein yield and the enrichment of non-essential amino acids compared to culture in unmodified, serum-free medium. Adding dichloroacetate, a pyruvate dehydrogenase kinase inhibitor, further improves cell viability, recombinant protein yield, and isotope enrichment. We demonstrate the method by producing partially enriched recombinant Thy1 glycoprotein from Lec1 Chinese hamster ovary (CHO) cells using U-13C-glucose and 15N-glutamate as labeled precursors. This study suggests that uniformly 15N,13C-labeled recombinant proteins may be produced in cultured mammalian cells starting from a mixture of labeled essential amino acids, glucose, and glutamate.

  14. Bordetella avium Antibiotic Resistance, Novel Enrichment Culture, and Antigenic Characterization

    Beach, Nathan M; Thompson, Seth; Mutnick, Rachel; Brown, Lisa; Kettig, Gina; Puffenbarger, Robyn; Miyamoto, David; Temple, Louise

    2012-01-01

    Bordetella avium continues to be an economic issue in the turkey industry as the causative agent of bordetellosis, which often leads to serious secondary infections. This study presents a broad characterization of the antibiotic resistance patterns in this diverse collection of B. avium strains collected over the past thirty years. In addition, the plasmid basis for the antibiotic resistance was characterized. The antibiotic resistance pattern allowed the development of a novel enrichment cul...

  15. Use of γ-hexachlorocyclohexane as a terminal electron acceptor by an anaerobic enrichment culture

    Highlights: ► Use of γ-hexachlorocyclohexane as a terminal electron acceptor was demonstrated. ► H2 served as the electron donor for an enrichment culture that dechlorinated γ-HCH. ► H2 consumption for acetogenesis and methanogenesis stopped in HEPES media. ► Addition of vancomycin significantly slowed the rate of γ-HCH dechlorination. ► Previously identified chlororespiring microbes were not detected in the enrichment. - Abstract: The use of γ-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 γ-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 γ-HCH consumed (on a molar basis), respectively. The enrichment culture remained active with only hydrogen as the electron donor and γ-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 γ-HCH dechlorination, suggesting that a Gram-positive organism is responsible for the reduction of γ-HCH. Analysis of the γ-HCH dechlorinating enrichment culture did not detect any known chlororespiring genera, including Dehalobacter. In bicarbonate-buffered medium, reductive dechlorination of γ-HCH was accompanied by significant levels of acetogenesis as well as methanogenesis.

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

    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, reductive dechlorination of {gamma}-HCH was accompanied by significant levels of acetogenesis as well as methanogenesis.

  17. Anaerobic degradation of isobutyrate by methanogenic enrichment cultures and by a Desuifococcus multivorans strain

    Stieb, Marion; Schink, Bernhard

    1989-01-01

    Methanogenic enrichment cultures with isobuty-rate as sole source of carbon and energy were inoculated with sediment and sludge samples from freshwater and marine origin. Over more than 20 transfers, these cultures fermented 2 mol isobutyrate with 1 mol CO2 via an intermediate for-marion of n-butyrate to 4 mol acetate and 1 mol CH4. The primary isobutyrate-fermenting bacteria could not be purified. From one of the marine enrichment cultures, a sulfate-reducing bacterium was isolated which oxi...

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

    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.

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

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

    2009-01-01

    The aim of this study was to enrich, characterize and identify strict anaerobic extreme thermophilic hydrogen (H-2) producers from digested household solid wastes. A strict anaerobic extreme thermophilic H-2 producing bacterial culture was enriched from a lab-scale digester treating household...... wastes at 70 degrees C. The enriched mixed culture consisted of two rod-shaped bacterial members growing at an optimal temperature of 80 degrees C and an optimal pH 8.1. The culture was able to utilize glucose, galactose, mannose, xylose, arabinose, maltose, sucrose, pyruvate and glycerol as carbon...... 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....

  20. Ca2+ enrichment in culture medium potentiates effect of oligonucleotides.

    Hori, Shin-Ichiro; Yamamoto, Tsuyoshi; Waki, Reiko; Wada, Shunsuke; Wada, Fumito; Noda, Mio; Obika, Satoshi

    2015-10-30

    Antisense and RNAi-related oligonucleotides have gained attention as laboratory tools and therapeutic agents based on their ability to manipulate biological events in vitro and in vivo. We show that Ca(2+) enrichment of medium (CEM) potentiates the in vitro activity of multiple types of oligonucleotides, independent of their net charge and modifications, in various cells. In addition, CEM reflects in vivo silencing activity more consistently than conventional transfection methods. Microscopic analysis reveals that CEM provides a subcellular localization pattern of oligonucleotides resembling that obtained by unassisted transfection, but with quantitative improvement. Highly monodispersed nanoparticles ~100 nm in size are found in Ca(2+)-enriched serum-containing medium regardless of the presence or absence of oligonucleotides. Transmission electron microscopy analysis reveals that the 100-nm particles are in fact an ensemble of much smaller nanoparticles (ϕ ∼ 15 nm). The presence of these nanoparticles is critical for the efficient uptake of various oligonucleotides. In contrast, CEM is ineffective for plasmids, which are readily transfected via the conventional calcium phosphate method. Collectively, CEM enables a more accurate prediction of the systemic activity of therapeutic oligonucleotides, while enhancing the broad usability of oligonucleotides in the laboratory. PMID:26101258

  1. Quantification of anaerobic ammonium-oxidizing bacteria in enrichment cultures by real-time PCR

    Tsushima, Ikuo; Kindaichi, Tomonori; Okabe, Satoshi

    2007-01-01

    The anaerobic ammonium-oxidizing (ANAMMOX) bacteria were enriched from a rotating disk reactor (RDR) biofilm in semi batch cultures. Based on fluorescence in situ hybridization (FISH) analysis, this enrichment led to a relative population size of 36% ANAMMOX bacteria. Phylogenetic analysis revealed that all the detected clones were related to the previously reported ANAMMOX bacteria, Candidatus Brocadia anammoxidans (AF375994), with 92% sequence similarity. Furthermore, we successfully develo...

  2. Transformations of Halogenated Aromatic Aldehydes by Metabolically Stable Anaerobic Enrichment Cultures

    Neilson, Alasdair H.; Allard, Ann-Sofie; Hynning, Per-ke; Remberger, Mikael

    1988-01-01

    Metabolically stable enrichment cultures of anaerobic bacteria obtained by elective enrichment of sediment samples from the Baltic Sea and Gulf of Bothnia have been used to study the oxidation and reduction of the aldehyde group of various halogenated aromatic aldehydes. During the transformation of 5- and 6-chlorovanillin, 6-bromovanillin, 3-chloro-4-hydroxybenzaldehyde, 3,5-dichloro-4-hydroxybenzaldehyde, and 3,5-dibromo-4-hydroxybenzaldehyde, it was shown that synthesis of the correspondin...

  3. Resource recovery from organic waste streams by microbial enrichment cultures:

    Tamis, J.

    2015-01-01

    Polyhydroxyalkanoate (PHA) is a natural product that can potentially replace a part of the chemicals and plastics derived from fossil sources. One of the main barriers for market entry of PHA is its relatively high price compared to conventional (fossil) feedstocks. This high price is related to current industrial production methods which are based on the cultivation of pure microbial cultures of a single species that a.o. has to be protected from contaminations from unwanted microorganisms t...

  4. Resource recovery from organic waste streams by microbial enrichment cultures

    Tamis, J.

    2015-01-01

    Polyhydroxyalkanoate (PHA) is a natural product that can potentially replace a part of the chemicals and plastics derived from fossil sources. One of the main barriers for market entry of PHA is its relatively high price compared to conventional (fossil) feedstocks. This high price is related to current industrial production methods which are based on the cultivation of pure microbial cultures of a single species that a.o. has to be protected from contaminations from unwanted microorganisms t...

  5. Two-stage mineralization of phenanthrene by estuarine enrichment cultures

    The polycyclic aromatic hydrocarbon phenanthrene was mineralized in two stages by soil, estuarine water, and sediment microbial populations. At high concentrations, phenanthrene was degraded, with the concomitant production of biomass and accumulation of Folin-Ciocalteau-reactive aromatic intermediates. Subsequent consumption of these intermediates resulted in a secondary increase in biomass. Analysis of intermediates by high-performance liquid chromatography, thin-layer chromatography, and UV absorption spectrometry showed 1-hydroxy-2-naphthoic acid (1H2NA) to be the predominant product. A less pronounced two-stage mineralization pattern was also observed by monitoring 14CO2 production from low concentrations (0.5 mg liter-1) of radiolabeled phenanthrene. Here, mineralization of 14C-labeled 1H2NA could explain the incremental 14CO2 produced during the later part of the incubations. Accumulation of 1H2NA by isolates obtained from enrichments was dependent on the initial phenanthrene concentration. The production of metabolites during polycyclic aromatic hydrocarbon biodegradation is discussed with regard to its possible adaptive significance and its methodological implications

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

    Palomeras, Sònia; Rabionet, Marc; Ferrer, Inés; Sarrats, Ariadna; Garcia-Romeu, Maria Luisa; Puig, Teresa; Ciurana, Joaquim

    2016-01-01

    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. PMID:27120585

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

    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.

  8. Anaerobic benzene degradation by iron- and sulfate-reducing enrichment cultures

    Abu Laban, Nidal

    2010-01-01

    Anaerobic benzene degradation was studied with a sulfate- and an iron-reducing enrichment culture (BPL and BF). In culture BPL, molecular analysis revealed 95% dominance of a novel phylotype that is affiliated to the genus Pelotomaculum. To elucidate the initial activation mechanism of anaerobic benzene degradation, putative metabolites were screened revealing benzoate as a product of a carboxylation reaction. For identification of enzymes involved in anaerobic benzene degradation the whole p...

  9. BACTERIOLOGY OF BODY FLUIDS WITH AN EVALUATION OF ENRICHMENT TECHNIQUE TO INCREASE CULTURE POSITIVITY

    Anasua

    2014-12-01

    Full Text Available BACKGROUND: Body fluids like ascitic fluids, pleural fluids, cerebrospinal fluids (CSF etc. are sent for culture in a clinical microbiology laboratory to achieve etiological diagnosis. However the yield of such cultures is usually very low. So, ongoing monitoring of prevalent pathogenic organisms and their sensitivities help the clinicians institute therapy in absence of a culture report. AIMS: The study was done to identify the common pathogens isolated from body fluids along with their antimicrobial susceptibility pattern and also to evaluate the impact of enrichment on their culture positivity. SETTING AND DESIGN: A 3-month prospective analytical study was done in a tertiary care hospital. MATERIALS AND METHODS: A total of 333 Body fluids were processed; 103 of them were ascitic fluids, 71 pleural fluids, 139 CSF and 20 other fluids. They were processed by plating the direct sample and after enrichment. Enrichment was done by two methods: in Soyabean-Caesin digest broth (274 samples and by BACTEC (59 samples.Isolates were identified by routine procedures & their antimicrobial susceptibility determined as per CLSI guidelines. The results were analyzed using Microsoft Excel® software using p<0.05 as the cut-off for significance. RESULTS: Gram negative isolate were obtained from 21.3% of the samples. The common isolates were Pseudomonas (20.7%, Acinetobacter (11.6%, Citrobacter (10.7% and E. coli (10.7%. The antibiotics most effective against Gram negative pathogens were Gentamicin (47.5%, Pipercillin-Tazobactam (51.6%, Amikacin (56.7% and Cefoperazone-Sulbactam(65.3%. Gram positive isolates, obtained from 9% of the samples, mostly consisted of MSSA, Enterococcus and CONS, for which Ciprofloxacin (48% followed by Cotrimoxazole (40% and Erythromycin (28.6% showed reasonable efficacy. The Culture positivity with direct plating, Soyabean-Caesin broth enrichment and BACTEC was 14.41%, 29.19% and 42.37% respectively. Increase in positivity by Soyabean-Casein broth was maximum for pleural fluids (12% followed by ascitic fluids (11.6% and CSF (11.52%.Using automated system the corresponding increases were 20.7%for ascitic fluids and 5.4%for pleural fluids. The mean time for identification using direct plating, enrichment method and BACTEC were 48 hours, 72 hours and 40 hours respectively. CONCLUSION: Gram negative isolates are commonly isolated pathogens from body fluids in our setup. Enrichment of body fluids improved yield of pathogens. In resource-poor settings simple enrichment in blood culture bottles can increase culture positivity of these precious samples.

  10. Summary report on the aerobic degradation of diesel fuel and the degradation of toluene under aerobic, denitrifying and sulfate reducing conditions

    This report contains a number of studies that were performed to better understand the technology of the biodegradation of petroleum hydrocarbons. Topics of investigation include the following: diesel fuel degradation by Rhodococcus erythropolis; BTEX degradation by soil isolates; aerobic degradation of diesel fuel-respirometry; aerobic degradation of diesel fuel-shake culture; aerobic toluene degradation by A3; effect of HEPES, B1, and myo-inositol addition on the growth of A3; aerobic and anaerobic toluene degradation by contaminated soils; denitrifying bacteria MPNs; sulfate-reducing bacteria MPNs; and aerobic, DNB and SRB enrichments

  11. Stable Isotopic Studies of n-Alkane Metabolism by a Sulfate-Reducing Bacterial Enrichment Culture

    Davidova, Irene A.; Lisa M. Gieg; Nanny, Mark; Kropp, Kevin G.; Suflita, Joseph M.

    2005-01-01

    Gas chromatography-mass spectrometry and nuclear magnetic resonance spectroscopy were used to study the metabolism of deuterated n-alkanes (C6 to C12) and 1-13C-labeled n-hexane by a highly enriched sulfate-reducing bacterial culture. All substrates were activated via fumarate addition to form the corresponding alkylsuccinic acid derivatives as transient metabolites. Formation of d14-hexylsuccinic acid in cell extracts from exogenously added, fully deuterated n-hexane confirmed that this reac...

  12. Degradation of unsaturated hydrocarbons by methanogenic enrichment cultures : (Aromatic compounds; alcohol degradation; hexadecene; squalene; hydratation; carboxylation)

    Schink, Bernhard

    1985-01-01

    The biodegradability of hydrocarbons under anaerobic conditions was studied in enrichment cultures using mineral media inoculated with sewage sludge or sediment samples of limnic and marine origin. No indication of methanogenic degradation was obtained with either n-hexane, n-hexadecane, n-heptadecane, 1-hexene, cis-2-hexene, trans-2-hexene, isoprene, 1-hexine, benzene, toluene, xylene, cyclohexene, cycloheptatriene, cyclopentadiene, styrene, naphthalene, azulene, or fl-carotene. Squalene was...

  13. Metabolic by-products of anaerobic toluene degradation by sulfate-reducing enrichment cultures.

    Beller, H. R.; Reinhard, M.; Grbić-Galić, D

    1992-01-01

    Two dead-end metabolites of anaerobic toluene transformation, benzylsuccinic acid and benzylfumaric acid, accumulated in sulfate-reducing enrichment cultures that were fed toluene as the sole carbon source. Stable isotope-labeled toluene and gas chromatography-mass spectrometry were used to confirm that the compounds resulted from toluene metabolism. The two metabolites constituted less than 10% of the toluene carbon (over 80% was mineralized to carbon dioxide, according to a previous study)....

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

    Trably, Eric; Batstone, Damien J.; Christensen, Nina; Patureau, Dominique; Schmidt, Jens Ejbye

    2008-01-01

    Although anaerobic biodegradation of di-n-butyl phthalic acid ester (DBP) has been studied over the past decade, only little is known about the microorganisms involved in the biological anaerobic degradation pathways. The aim of this work is to characterize the microbial community dynamics in...... losses were observed in the sterile controls (20-22%), substantial DBP biodegradation was found in the enrichment cultures (90-99%). In addition, significant population changes were observed. The dominant bacterial species in the DBP-degrading cultures was affiliated to Soehngenia saccharolytica, a...

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

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

    2015-09-01

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

  16. Identification of active denitrifiers in full-scale nutrient removal wastewater treatment systems.

    McIlroy, Simon Jon; Starnawska, Anna; Starnawski, Piotr; Saunders, Aaron Marc; Nierychlo, Marta; Nielsen, Per Halkjaer; Nielsen, Jeppe Lund

    2016-01-01

    Denitrification is essential to the removal of nitrogen from wastewater during treatment, yet an understanding of the diversity of the active denitrifying bacteria responsible in full-scale wastewater treatment plants (WWTPs) is lacking. In this study, stable-isotope probing (SIP) was applied in combination with microautoradiography (MAR)-fluorescence in situ hybridization (FISH) to identify previously unrecognized active denitrifying phylotypes in a full-scale WWTP with biological N and P removal. Acknowledging that different denitrifiers will have specific carbon source preferences, a fully (13)C-labelled complex substrate was used for SIP incubations, under nitrite-reducing conditions, in order to maximize the capture of the potentially metabolically diverse denitrifiers likely present. Members of the Rhodoferax, Dechloromonas, Sulfuritalea, Haliangium and Thermomonas were represented in the 16S rRNA gene clone libraries from DNA enriched in (13)C, with FISH probes optimized here for their in situ characterization. FISH and MAR confirmed that they were all active denitrifiers in the community. The combined approach of SIP and MAR-FISH represents an excellent approach for identifying and characterizing an un-described diversity of active denitrifiers in full-scale systems. PMID:25181571

  17. Biodegradation of munitions compounds by a sulfate reducing bacterial enrichment culture

    Boopathy, R.; Manning, J. [Argonne National Lab., IL (United States). Environmental Research Div.

    1997-08-01

    The degradation of several munitions compounds was studied. The compounds included 2,4,6-trinitrotoluene (TNT), hexahydro-1,3,5-trinitro-1,3,5-triazine, octahydro-1,3,5,7-tetranitro-1,3,5,7-tetraazocine, 2,4,6-trinitrobenzene (TNB), and 2,4-dinitrotoluene. All of the compounds studied were degraded by the sulfate reducing bacterial (SRB) enrichment culture. The SRB culture did not use the munitions compounds as their sole source of carbon. However, all the munitions compounds tested served as the sole source of nitrogen for the SRB culture. Degradation of munitions compounds was achieved by a co-metabolic process. The SRB culture used a variety of carbon sources including pyruvate, ethanol, formate, lactate, and H{sub 2}-CO{sub 2}. The SRB culture was an incomplete oxidizer, unable to carry out the terminal oxidation of organic substrates to CO{sub 2} as the sole product, and it did not use acetate or methanol as a carbon source. In addition to serving as nitrogen sources, the munitions compounds also served as electron acceptors in the absence of sulfate. A soil slurry experiment with 5% and 10% munitions compounds-contaminated soil showed that the contaminant TNT was metabolized by the SRB culture in the presence of pyruvate as electron donor. This culture may be useful in decontaminating munitions compounds-contaminated soil and water under anaerobic conditions.

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

    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.

  19. Tetrachloromethane-Degrading Bacterial Enrichment Cultures and Isolates from a Contaminated Aquifer

    Christian Penny

    2015-07-01

    Full Text Available The prokaryotic community of a groundwater aquifer exposed to high concentrations of tetrachloromethane (CCl4 for more than three decades was followed by terminal restriction fragment length polymorphism (T-RFLP during pump-and-treat remediation at the contamination source. Bacterial enrichments and isolates were obtained under selective anoxic conditions, and degraded 10 mgL?1 CCl4, with less than 10% transient formation of chloroform. Dichloromethane and chloromethane were not detected. Several tetrachloromethane-degrading strains were isolated from these enrichments, including bacteria from the Klebsiella and Clostridium genera closely related to previously described CCl4 degrading bacteria, and strain TM1, assigned to the genus Pelosinus, for which this property was not yet described. Pelosinus sp. TM1, an oxygen-tolerant, Gram-positive bacterium with strictly anaerobic metabolism, excreted a thermostable metabolite into the culture medium that allowed extracellular CCl4 transformation. As estimated by T-RFLP, phylotypes of CCl4-degrading enrichment cultures represented less than 7%, and archaeal and Pelosinus strains less than 0.5% of the total prokaryotic groundwater community.

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

    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. PMID:25656248

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

    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...

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

    Angelidaki, I.; Ahring, B K

    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 culture consisted of a short rod and two bacteria antigenically related to Methanobacterium thermoautotrophicum (Delta)H and Methanosarcina thermophila TM-1.

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

    Carvajal-Arroyo, Jos 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. PMID:23245574

  4. Halobacterium denitrificans sp. nov., an extremely halophilic denitrifying bacterium

    Tomlinson, G. A.; Jahnke, L. L.; Hochstein, L. I.

    1986-01-01

    Halobacterium denitrificans was one of several carbohydrate-utilizing, denitrifying, extremely halophilic bacteria isolated by anaerobic enrichment in the presence of nitrate. Anaerobic growth took place only when nitrate (or nitrite) was present and was accompanied by the production of dinitrogen. In the presence of high concentrations of nitrate (i.e., 0.5 percent), nitrous oxide and nitrite were also detected. When grown aerobically in a mineral-salts medium containing 0.005 percent yeast extract, H. denitrificans utilized a variety of carbohydrates as sources of carbon and energy. In every case, carbohydrate utilization was accompanied by acid production.

  5. Halobacterium denitrificans sp. nov. - An extremely halophilic denitrifying bacterium

    Tomlinson, G. A.; Jahnke, L. L.; Hochstein, L. I.

    1986-01-01

    Halobacterium denitrificans was one of several carbohydrate-utilizing, denitrifying, extremely halophilic bacteria isolated by anaerobic enrichment in the presence of nitrate. Anaerobic growth took place only when nitrate (or nitrite) was present and was accompanied by the production of dinitrogen. In the presence of high concentrations of nitrate (i.e., 0.5 percent), nitrous oxide and nitrite were also detected. When grown aerobically in a mineral-salts medium containing 0.005 percent yeast extract, H. denitrificans utilized a variety of carbohydrates as sources of carbon and energy. In every case, carbohydrate utilization was accompanied by acid production.

  6. Large scale production of Blackleg vaccine by fermenter and enriched culture medium in Iran

    Pilehchian Langroudi, R.

    2012-06-01

    Full Text Available In all biological systems growth is defined as increase of chemical compounds. Bacteria can achieve to balanced growth if they are growing in a medium, which are completely adapted to it. Clostridium chauvoei, (Clostridium feseri is an anaerobic, spore forming, motile, and polymorph bacteria, which its size varies from 0.5-1 to 3-8 micron and could be observed as individual bacterium, diplo, and rarely streptococcus. Blackleg is a fatal disease of young cattle. It produces an acute local infection, and the resulting blood poisoning leads to rapid death. Clostridium chauvoei and, less frequently, Clostridium septicum are the most commonly responsible organisms. Vaccination is the only effective means for controlling of blackleg disease. Several kinds of vaccine are available commercially. It is 4 decades that blackleg vaccine is produced in Razi institute and because of enhanced demand of country, decision was made to improve the production procedure of this vaccine using large-scale fermenter, so the aim of this study was adaptation of Clostridium chauvoei to growth and proliferation in fermenter for preparation of a potent vaccine. Accordingly attempts were made to prepare and formulate the ingredients in order to obtain high yield of Clostridium chauvoei in culture medium by fermenter. All experiments were done in two sets: A-growth in glass bottles using conventional culture medium and B-growth in fermenter using conventional culture medium similar to A and also enriched culture medium. Results showed high yield of Clostridium chauvoei suspension in fermenter after 10 hours, using enriched culture medium (more than 1,480,000,000 organisms/ml, but no significant changes was obtained in glass bottles conditions comparing to the fermenter conditions. The safety and potency of the prepared vaccine was determined in sheep and guinea pigs according to British pharmacopoeia (veterinary with satisfactory results. Since this research has been successfully done in Razi research institute, so the mono valent inactivated blackleg vaccine, using the enriched culture medium is currently producing by fermenter and is used for immunization of cattles in Iran.

  7. Who is actively denitrifying in activated sludge?

    Hansen, Aviaja Anna; Nielsen, Jeppe Lund

    Bacteroidetes. Furthermore, potential denitrifying genera of Alpha- and Betaproteobacteria were quantified in the activated sludge with 16S rRNA gene probes for fluorescence in situ hybridization (FISH). This revealed that Aquaspirillum-related bacteria were dominant followed by bacteria related to Azoarcus......Denitrification is of crucial importance in nitrogen removal from wastewater. However, due to the polyphyletic taxonomy of denitrifiers, little is known about the composition and ecophysiology of the actively denitrifying community in activated sludge. To identify the active denitrifiers in a full......-scale wastewater treatment plant the transcripts (mRNA) of the nirS, nirK and nosZ denitrification genes expressed under acetate or amino acid consumption were amplified, sequenced and identified. This revealed that the majority of the denitrifiers belonged to Alpha- and Betaproteobacteria, while only few...

  8. Paralytic shellfish toxin production by the dinoflagellate Alexandrium pacificum (Chinhae Bay, Korea) in axenic, nutrient-limited chemostat cultures and nutrient-enriched batch cultures.

    Han, Myungsoo; Lee, Haeok; Anderson, Donald M; Kim, Baikho

    2016-03-15

    Blooms of Alexandrium pacificum (formerly Alexandrium tamarense) are common in Chinhae Bay (Korea), presumably linked to anthropogenic eutrophication. Here we examine PSP toxin content and composition in axenic chemostat and batch cultures of A. pacificum using growth conditions that differed according to dilution rate, nutrient limitations, and enrichments. Phosphate (P)-limited cells in chemostat cultures had higher toxin content and a toxin composition that differed from that of nitrogen (N)-limited cells at the highest growth rates. Therefore, toxin composition changes do occur in axenic cultures of A. pacificum following extended growth under steady state conditions. In nutrient-limited batch cultures that received N and P enrichment, the N-enriched cultures showed a more diverse toxin profile than the P-enriched cells; the toxin content of N-enriched cells was lower than in the P-enriched cultures. We infer the following order for the biosynthesis of individual toxins: C1, C2>GTX3>GTX1>neoSTX. PMID:26874747

  9. Exploration and isolation of novel thermophiles in frozen enrichment cultures derived from a terrestrial acidic hot spring.

    Sakai, Hiroyuki D; Kurosawa, Norio

    2016-03-01

    An isolation strategy, exploring novel microorganisms in frozen enrichment cultures (ENFE), which uses a combination of enrichment culture and 16S rRNA gene clone analysis, was evaluated for isolating uncultured thermophiles from a terrestrial acidic hot spring. The procedure comprised (a) multiple enrichment cultures under various conditions, (b) cryostorage of all enrichments, (c) microbial community analyses of the enrichments using 16S rRNA gene sequences, and (d) purification of microorganisms from enrichments containing previously uncultured microorganisms. The enrichments were performed under a total of 36 conditions, and 16 of these enrichments yielded positive microbial growth with the detection of three previously uncultured archaea. Two of the three previously uncultured archaea, strains HS-1 and HS-3, were successfully isolated. Strain HS-1 and HS-3 represented a novel lineage of the order Sulfolobales and novel species of the genus Sulfolobus, respectively. Although innovative isolation methods play strategic roles in isolating previously uncultured microorganisms, the ENFE strategy showed potential for characterizing and isolating such microorganisms using conventional media and techniques. PMID:26860120

  10. Comparison of bacterial enriched-broth culture, enzyme linked immunosorbent assay, and broth culture-polymerase chain reaction techniques for identifying asymptomatic infections with Salmonella in swine

    Sibley, Jennifer; Yue, Binbin; Huang, Fei; Harding, John; Kingdon, Jill; Chirino-Trejo, Manuel; Appleyard, Greg D.

    2003-01-01

    A polymerase chain reaction (PCR) assay was combined with a broth-culture enrichment system to detect Salmonella shed in feces from subclinically infected swine. The effectiveness of the broth culture-polymerase chain reaction (BC-PCR) assay to identify pigs shedding Salmonella in feces was compared with a microbiological culture and a commercial enzyme linked immunosorbent assay (ELISA) kit to detect Salmonella-specific serum antibody. A total of 67 pigs were tested by each of the 3 methodol...

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

    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. PMID:26832872

  12. Bacterial oxidation of dibromomethane and methyl bromide in natural waters and enrichment cultures

    Goodwin, K.D.; Schaefer, J.K.; Oremland, R.S.

    1998-01-01

    Bacterial oxidation of 14CH2Br2 and 14CH3Br was measured in freshwater, estuarine, seawater, and hypersaline-alkaline samples. In general, bacteria from the various sites oxidized similar amounts of 14CH2Br2 and comparatively less 14CH3Br. Bacterial oxidation of 14CH3Br was rapid in freshwater samples compared to bacterial oxidation of 14CH3Br in more saline waters. Freshwater was also the only site in which methyl fluoride-sensitive bacteria (e.g., methanotrophs or nitrifiers) governed brominated methane oxidation. Half-life calculations indicated that bacterial oxidation of CH2Br2 was potentially significant in all of the waters tested. In contrast, only in freshwater was bacterial oxidation of CH3Br as fast as chemical removal. The values calculated for more saline sites suggested that bacterial oxidation of CH3Br was relatively slow compared to chemical and physical loss mechanisms. However, enrichment cultures demonstrated that bacteria in seawater can rapidly oxidize brominated methanes. Two distinct cultures of nonmethanotrophic methylotrophs were recovered; one of these cultures was able to utilize CH2Br2 as a sole carbon source, and the other was able to utilize CH3Br as a sole carbon source.

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

    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 meta...

  14. Continuous enrichment culture and molecular monitoring to investigate the microbial diversity of thermophiles inhabiting deep-sea hydrothermal ecosystems.

    Postec, Anne; Urios, Laurent; Lesongeur, Françoise; Ollivier, Bernard; Querellou, Joël; Godfroy, Anne

    2005-03-01

    The microflora developing during a continuous enrichment culture from a hydrothermal chimney sample was investigated by molecular methods. The culture was performed in a gas-lift bioreactor under anaerobic conditions, at 90 degrees C and pH 6.5, on a complex medium containing sulfur as the terminal electron acceptor. Archaeal and bacterial diversity was studied. Microorganisms affiliated with the genera Pyrococcus, Marinitoga, and Bacillus were detected through DGGE analysis of 16S rDNA. Additional sequences phylogenetically related to Thermococcus and epsilon-Proteobacteria were detected by cloning and sequencing of 16S rDNA from two samples of the enrichment culture. In comparison, the sequences retrieved from cloning analysis from an enrichment culture performed in a flask (batch condition) using the same culture medium showed that only members of the genus Thermococcus were cultivated. Therefore, continuous enrichment culture using the gas-lift bioreactor can be considered as an efficient and improved method for investigating microbial communities originating from deep-sea hydrothermal vents. PMID:15717222

  15. Continuous enrichment culturing of thermophiles under sulfate and nitrate-reducing conditions and at deep-sea hydrostatic pressures.

    Houghton, J L; Seyfried, W E; Banta, A B; Reysenbach, A-L

    2007-03-01

    A continuous culture bioreactor was developed to enrich for nitrate and sulfate reducing thermophiles under in situ deep-sea pressures. The ultimate objective of this experimental design was to be able to study microbial activities at chemical and physical conditions relevant to seafloor hydrothermal vents. Sulfide, sulfate and oxide minerals from sampled seafloor vent-chimney structures [East Pacific Rise (9 degrees 46'N)] served as source mineral and microbial inoculum for enrichment culturing using nitrate and sulfate-enriched media at 70 and 90 degrees C and 250 bars. Changes in microbial diversity during the continuous reaction flow were monitored using denaturing gradient gel electrophoresis (DGGE) of PCR amplified 16S rRNA gene fragments. Time series changes in fluid chemistry were also monitored throughout the experiment to assess the feedback between mineral-fluid reaction and metabolic processes. Data indicate a shift from the dominance of epsilon Proteobacteria in the initial inoculum to the several Aquificales-like phylotypes in nitrate-reducing enrichment media and Thermodesulfobacteriales in the sulfate-reducing enrichment media. Methanogens were detected in the original sulfide sample and grew in selected sulfate-enriched experiments. Microbial interactions with anhydrite and pyrrhotite in the chimney material resulted in measurable changes in fluid chemistry despite a fluid residence time only 75 min in the reactor. Changes in temperature rather than source material resulted in greater differences in microbial enrichments and mediated geochemical reactions. PMID:17221162

  16. Denitrifying Bacteria in the Earthworm Gastrointestinal Tract and In Vivo Emission of Nitrous Oxide (N(inf2)O) by Earthworms

    Karsten, G. R.; H. L. Drake

    1997-01-01

    Earthworms (Lumbricus rubellus and Octolasium lacteum) and gut homogenates did not produce CH(inf4), and methanogens were not readily culturable from gut material. In contrast, the numbers of culturable denitrifiers averaged 7 x 10(sup7) and 9 x 10(sup6) per g (dry weight) of gut material for L. rubellus and O. lacteum, respectively; these values were 256- and 35-fold larger than the numbers of culturable denitrifiers in the soil from which the earthworms were obtained. Anaerobically incubate...

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

    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. PMID:23892419

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

    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)

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

    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 etal. (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 (4h 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. PMID:26187833

  20. Anaerobic Degradation of Benzene, Toluene, Ethylbenzene, and o-Xylene in Sediment-Free Iron-Reducing Enrichment Cultures

    Jahn, Michael K.; Haderlein, Stefan B.; Meckenstock, Rainer U.

    2005-01-01

    Monoaromatic hydrocarbons such as benzene, toluene, ethylbenzene, and xylene (BTEX) are widespread contaminants in groundwater. We examined the anaerobic degradation of BTEX compounds with amorphous ferric oxide as electron acceptor. Successful enrichment cultures were obtained for all BTEX substrates both in the presence and absence of AQDS (9,10-anthraquinone-2,6-disulfonic acid). The electron balances showed a complete anaerobic oxidation of the aromatic compounds to CO2. This is the first report on the anaerobic degradation of o-xylene and ethylbenzene in sediment-free iron-reducing enrichment cultures. PMID:15933041

  1. High yield derivation of enriched glutamatergic neurons from suspension-cultured mouse ESCs for neurotoxicology research

    Hubbard Kyle S

    2012-10-01

    Full Text Available Abstract Background Recently, there has been a strong emphasis on identifying an in vitro model for neurotoxicity research that combines the biological relevance of primary neurons with the scalability, reproducibility and genetic tractability of continuous cell lines. Derived neurons should be homotypic, exhibit neuron-specific gene expression and morphology, form functioning synapses and consistently respond to neurotoxins in a fashion indistinguishable from primary neurons. However, efficient methods to produce neuronal populations that are suitable alternatives to primary neurons have not been available. Methods With the objective of developing a more facile, robust and efficient method to generate enriched glutamatergic neuronal cultures, we evaluated the neurogenic capacity of three mouse embryonic stem cell (ESC lines (R1, C57BL/6 and D3 adapted to feeder-independent suspension culture. Neurogenesis and neuronal maturation were characterized as a function of time in culture using immunological, genomic, morphological and functional metrics. The functional responses of ESNs to neurotropic toxins with distinctly different targets and mechanisms of toxicity, such as glutamate, α-latrotoxin (LTX, and botulinum neurotoxin (BoNT, were also evaluated. Results Suspension-adapted ESCs expressed markers of pluripotency through at least 30 passages, and differentiation produced 97×106 neural progenitor cells (NPCs per 10-cm dish. Greater than 99% of embryonic stem cell-derived neurons (ESNs expressed neuron-specific markers by 96 h after plating and rapidly developed complex axodendritic arbors and appropriate compartmentalization of neurotypic proteins. Expression profiling demonstrated the presence of transcripts necessary for neuronal function and confirmed that ESN populations were predominantly glutamatergic. Furthermore, ESNs were functionally receptive to all toxins with sensitivities and responses consistent with primary neurons. Conclusions These findings demonstrate a cost-effective, scalable and flexible method to produce a highly enriched glutamatergic neuron population. The functional characterization of pathophysiological responses to neurotropic toxins and the compatibility with multi-well plating formats were used to demonstrate the suitability of ESNs as a discovery platform for molecular mechanisms of action, moderate-throughput analytical approaches and diagnostic screening. Furthermore, for the first time we demonstrate a cell-based model that is sensitive to all seven BoNT serotypes with EC50 values comparable to those reported in primary neuron populations. These data providing compelling evidence that ESNs offer a neuromimetic platform suitable for the evaluation of molecular mechanisms of neurotoxicity.

  2. Continuous enrichment culture and molecular monitoring to investigate the microbial diversity of thermophiles inhabiting deep-sea hydrothermal ecosystems

    POSTEC, Anne; Urios, Laurent; Lesongeur, Francoise; Ollivier, Bernard; Querellou, Joel; Godfroy, Anne

    2005-01-01

    The microflora developing during a continuous enrichment culture from a hydrothermal chimney sample was investigated by molecular methods. The culture was performed in a gas-lift bioreactor under anaerobic conditions, at 90 degrees C and pH 6.5, on a complex medium containing sulfur as the terminal electron acceptor. Archaeal and bacterial diversity was studied. Microorganisms affiliated with the genera Pyrococcus, Marinitoga, and Bacillus were detected through DGGE analysis of 16S rDNA. Addi...

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

    Jensen, Annette Nygaard; Srensen, Gitte; Baggesen, Dorte Lau; Bdker, Rene; Hoorfar, Jeffrey

    2003-01-01

    , 90 samples opposed to 50 yielded a strong positive reading (+++) when Novobiocin was applied. Growth was on average 0.3 scores higher when pre-enriched with Novobiocin. The difference in growth score medians of the two methods was highly significant (Sign test; p <0.001). Despite the increased...... pre-enrichment broth with BPW made semi-selective by addition of Novobiocin. The semi-quantitative assessment of the Salmonella level showed that addition of Novobiocin in the pre-enrichment step on average increased the level of Salmonella 1.2 log dilution steps. When growth was scored at five levels...... sensitivity, 13 culture-positive samples were missed when using the Novobiocin-containing BPW. In conclusion, a simple addition of Novobiocin in the BPW pre-enrichment step of fecal samples may facilitate reading and thereby detection of Salmonella on MSRV. The increase of Salmonella in the semi...

  4. Chronic impact of tetracycline on nitrification kinetics and the activity of enriched nitrifying microbial culture.

    Katipoglu-Yazan, Tugce; Merlin, Christophe; Pons, Marie-Nolle; Ubay-Cokgor, Emine; Orhon, Derin

    2015-04-01

    This study evaluated the chronic impact of tetracycline on biomass with enriched nitrifying community sustained in a lab-scale activated sludge system. For this purpose, a fill and draw reactor fed with 100mg COD/L of peptone mixture and 50mgN/L of ammonia was sustained at a sludge age of 15 days. At steady-state, the reactor operation was continued with a daily tetracycline dosing of 50mg/L for more than 40 days, with periodic monitoring of the microbial composition, the nitrifying bacteria abundance, as well as the amoA and 16S rRNA gene activity, using molecular techniques. Changes in the kinetics of nitrification were quantified by modelling concentration profiles of major nitrogen fractions and oxygen uptake rate profiles derived from parallel batch experiments. Activated sludge modeling results indicated inhibitory impact of tetracycline on the growth of nitrifiers with a significant increase of the half saturation coefficients in corresponding rate equations. Tetracycline also inactivated biomass components of the enriched culture at a gradually increasing rate with time of exposure, leading to total collapse of nitrification. Molecular analyses revealed significant changes in the composition of the microbial community throughout the observation period. They also showed that continuous exposure to tetracycline inflicted significant reduction in amoA mRNA and 16S rRNA levels directly affecting nitrification. The chronic impact was much more pronounced on the ammonia oxidizing bacteria (AOB) community. These observations explained the basis of numerical changes identified in the growth kinetics of nitrifiers under stress conditions. PMID:25616640

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

    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??M) or 10x (700??M) 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. PMID:25978545

  6. Draft Genome Sequence of Alteromonas Macleodii Strain MIT1002, Isolated from an Enrichment Culture of the Marine Cyanobacterium Prochlorococcus

    Coe, Allison; Martin-Cuadrado, Ana-Belen; Biller, Steven; Chisholm, Sallie

    2015-01-01

    Alteromonas spp. are heterotrophic gammaproteobacteria commonly found in marine environments. We present here the draft genome sequence of Alteromonas macleodii MIT1002, which was isolated from an enrichment culture of the marine cyanobacterium Prochlorococcus NATL2A. This genome contains a mixture of features previously seen only within either the “surface” or “deep” Alteromonas ecotype.

  7. Draft Genome Sequence of Alteromonas macleodii Strain MIT1002, Isolated from an Enrichment Culture of the Marine Cyanobacterium Prochlorococcus

    Biller, Steven J.; Coe, Allison; Martin-Cuadrado, Ana-Belen; Chisholm, Sallie W

    2015-01-01

    Alteromonas spp. are heterotrophic gammaproteobacteria commonly found in marine environments. We present here the draft genome sequence of Alteromonas macleodii MIT1002, which was isolated from an enrichment culture of the marine cyanobacterium Prochlorococcus NATL2A. This genome contains a mixture of features previously seen only within either the “surface” or “deep” Alteromonas ecotype.

  8. Functional gene pyrosequencing and network analysis: an approach to examine the response of denitrifying bacteria to increased nitrogen supply in salt marsh sediments

    JenniferL.Bowen

    2013-11-01

    Full Text Available Functional gene pyrosequencing is emerging as a useful tool to examine the diversity and abundance of microbes that facilitate key biogeochemical processes. One such process, denitrification, is of particular importance because it converts fixed nitrate (NO3- to N2 gas, which returns to the atmosphere. In N limited salt marshes, removal of NO3- prior to entering adjacent waters helps prevent eutrophication. Understanding the dynamics of salt marsh microbial denitrification is thus imperative for the maintenance of healthy coastal ecosystems. We used pyrosequencing of the nirS gene to examine the denitrifying community response to fertilization in experimentally enriched marsh plots. A key challenge in the analysis of sequence data derived from pyrosequencing is understanding whether small differences in gene sequences are ecologically meaningful. We apply a novel approach from information theory that determined that the optimal similarity level for clustering DNA sequences into OTUs, while still capturing the ecological complexity of the system, was 88% similarity. With this clustering, phylogenetic analysis yielded 6 dominant clades of denitrifiers, the largest of which, accounting for more than half of all the sequences collected, had no close cultured representatives. Of the 638 OTUs identified, only 11 were present in all plots and no single OTU was dominant. We did, however, find a large number of specialist OTUs that were present only in a single plot. The high degree of endemic OTUs, while accounting for a large proportion of the nirS diversity in the plots, were found in lower abundance than the generalist taxa. The proportion of specialist taxa increased with increasing supply of nutrients, suggesting that addition of fertilizer may create conditions that expand the niche space for denitrifying organisms and may enhance the genetic capacity for denitrification.

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

    LauraAudreyHug

    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.

  10. Macrofaunal Impact on the Denitrifying Bacterial Community in Freshwater Sediment

    Poulsen, Morten; Stief, Peter; Schramm, Andreas

    , with nitrate-reducing and denitrifying bacteria as examples. The diversity of these functional groups was compared in sediment microcosms with and without Chironomus plumosus larvae, using the genes encoding nitrate reductase (narG) and nitrous oxide reductase (nosZ) as functional markers. The...... estimated phylotype richness of narG increased from 68 in sediment without larvae to 170 in sediment with larvae. Part of this increase in narG diversity could be explained by metabolic activation of certain nitrate-reducing bacteria in the gut of C. plumosus, since 18.3 % of the additional phylotypes were...... enriching the sediment with partly degraded organic material excreted by the larvae. In contrast to narG, nosZ phylotype richness was unaffected by the presence of chironomid larvae, and very few nosZ phylotypes were actively expressed in the larvae gut. Our results suggest that burrowing macroinvertebrates...

  11. Passive markers as a low-cost method of enriching cultural visits on user’s demand

    Anestis Koutsoudis

    2014-01-01

    Full Text Available The idea of enriching a cultural heritage visitor’s experience by employing mobile devices such as smartphones and tablets is an active research domain. This is mainly due to the worldwide user penetration of such devices along with their continuous hardware performance enhancement. This work presents the experience and feedback gained by one year of operation of an experimental information system that aims in enhancing the experience of a visitor on his/her demand. The evaluated system is based on the use case-scenario where the a mobile device is used to capture and decode passive markers (such as QR-codes that are located on facades of selected important buildings in the old town of Xanthi, Greece, in order to enrich the visitor’s experience through a broad context of historical and architectural information delivered over the Web.   Keywords: On-demand information, Passive marker, QR-code, Smartphone, Visit enrichment.

  12. PCR amplification of Bartonella koehlerae from human blood and enrichment blood cultures

    Breitschwerdt Edward B

    2010-08-01

    Full Text Available Abstract Background Cats appear to be the primary reservoir host for Bartonella koehlerae, an alpha Proteobacteria that is most likely transmitted among cat populations by fleas (Ctenocephalides felis. Bartonella koehlerae has caused endocarditis in a dog and in one human patient from Israel, but other clinically relevant reports involving this bacterium are lacking. Despite publication of numerous, worldwide epidemiological studies designed to determine the prevalence of Bartonella spp. bacteremia in cats, B. koehlerae has never been isolated using conventional blood agar plates. To date, successful isolation of B. koehlerae from cats and from the one human endocarditis patient has consistently required the use of chocolate agar plates. Results In this study, Bartonella koehlerae bacteremia was documented in eight immunocompetent patients by PCR amplification and DNA sequencing, either prior to or after enrichment blood culture using Bartonella alpha Proteobacteria growth medium. Presenting symptoms most often included fatigue, insomnia, joint pain, headache, memory loss, and muscle pain. Four patients were also infected with Bartonella vinsonii subsp. berkhoffii genotype II. After molecular documentation of B. koehlerae infection in these patients, a serological test was developed and serum samples were tested retrospectively. Bartonella koehlerae antibodies were not detected (titers B. koehlerae antibody titers of 1:64 or greater. Conclusions Although biased by a study population consisting of individuals with extensive arthropod and animal exposure, the results of this study suggest that B. koehlerae bacteremia is more common in immunocompetent people than has been previously suspected. Future studies should more thoroughly define modes of transmission and risk factors for acquiring infection with B. koehlerae. In addition, studies are needed to determine if B. koehlerae is a cause or cofactor in the development of arthritis, peripheral neuropathies or tachyarrhythmias in patients.

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

    Lpez, Juan C; Quijano, Guillermo; Prez, Rebeca; Muoz, Ral

    2014-12-15

    Methanotrophic communities were enriched in three stirred tank reactors continuously supplied with CH4-laden air at 20, 2 and 0.2gCH4m(-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 2gCH4m(-3) presenting the highest qmax and those enriched at 20 and 0.2gCH4m(-3) exhibiting the lowest KS. Maximum polyhydroxybutyrate (PHB) contents of 1.0% and 12.6% (w/w) were achieved at 20 and 2g CH4m(-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 0.2gCH4m(-3), respectively. PMID:25169644

  14. Anaerobic Degradation of Benzene, Toluene, Ethylbenzene, and o-Xylene in Sediment-Free Iron-Reducing Enrichment Cultures

    Jahn, Michael K.; Haderlein, Stefan B.; Meckenstock, Rainer U.

    2005-01-01

    Monoaromatic hydrocarbons such as benzene, toluene, ethylbenzene, and xylene (BTEX) are widespread contaminants in groundwater. We examined the anaerobic degradation of BTEX compounds with amorphous ferric oxide as electron acceptor. Successful enrichment cultures were obtained for all BTEX substrates both in the presence and absence of AQDS (9,10-anthraquinone-2,6-disulfonic acid). The electron balances showed a complete anaerobic oxidation of the aromatic compounds to CO2. This is the first...

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

    LauraAudreyHug; ElizabethAnneEdwards

    2013-01-01

    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 dehaloge...

  16. Formation of spontaneous hematopoietic colonies in semisolid cultures of lymphocyte enriched fractions of peripheral blood from beagles

    The formation of hematopoietic colonies in semisolid cultures derived from lymphocyte enriched fractions of canine peripheral blood is described. Such colonies occurred in the absence of either mitogens or colony stimulating factor. Although the mechanism(s) involved in such colony formation is not known, preliminary results suggest an increased tendency for spontaneous colony formation with age and a possible relationship to low-level irradiation and neoplasia. The observation underlines the importance of doing nonmitogen stimulated control cultures in order to determine the true influence of mitogens or growth factors on colony formation

  17. Degradation of polycyclic aromatic hydrocarbons by microbial consortia enriched from three soils using two different culture media

    A consortium composed of many different bacterial species is required to efficiently degrade polycyclic aromatic hydrocarbons (PAH) in oil-contaminated soil. We obtained six PAH-degrading microbial consortia from three oil-contaminated soils using two different isolation culture media. Denaturing gradient gel electrophoresis (DGGE) and sequence analyses of amplified 16s rRNA genes confirmed the bacterial community was greatly affected by both the culture medium and the soil from which the consortia were enriched. Three bacterial consortia enriched using malt yeast extract (MYE) medium showed higher degradation rates of PAHs than consortia enriched using Luria broth (LB) medium. Consortia obtained from a soil and then added back to that same soil was more effective in degrading PAHs than adding, to the same soil, consortia isolated from other, unrelated soils. This suggests that inoculum used for bioremediation should be from the same, or very similar nearby soils, as the soil that is actually being bioremediated. -- Highlights: •Six PAH-degrading microbial consortia were isolated from three oil-contaminated soils. •The bacterial community by 16s rRNA genes was affected by culture media and source soil. •Inoculum should be from the same or similar soil as the soil being bioremediated. -- Bioremediation of oil-contaminated soils was most effective when using inoculum of microbial consortia from the same or similar soil as the soil being bioremediated

  18. Isolation of uncultivated anaerobic thermophiles from compost by supplementing cell extract of Geobacillus toebii in enrichment culture medium.

    Bae, Jin-Woo; Rhee, Sung-Keun; Park, Ja Ryeong; Kim, Byung-Chun; Park, Yong-Ha

    2005-12-01

    Several researchers have reported that microorganisms can be cultivated only in the presence of other microorganisms. We suggest that a portion of uncultivated microorganisms might be cultivated in the presence of cellular components released from bacteria in their natural environments. In this study, the cell extract of Geobacillus toebii was used to enrich uncultivated thermophiles from compost. In the process of enrichment cultures, cell extract supplementation apparently changed the community composition. This change was monitored by PCR-DGGE targeting 16S rRNA gene. Five novel groups of microorganisms (similarity of 16S rRNA gene to the closest relative <96%) were specifically isolated from enrichment cultures by using cell extract-supplemented culture media. Their growth was found to be dependent on the addition of extract of G. toebii. Putting these findings together, we suggest that the extracts of bacteria could be one of the growth factors in the thermal ecosystem with a possibility of extending other ecological niches. PMID:16041476

  19. Denitrifying bacteria in bulk and maize-rhizospheric soil: diversity and N2O-reducing abilities.

    Chèneby, D; Perrez, S; Devroe, C; Hallet, S; Couton, Y; Bizouard, F; Iuretig, G; Germon, J C; Philippot, L

    2004-07-01

    The aim of this study was to determine the effect of the rhizosphere of maize on the diversity of denitrifying bacteria. Community structure comparison was performed by constructing a collection of isolates recovered from bulk and maize planted soil. A total of 3240 nitrate-reducing isolates were obtained and 188 of these isolates were identified as denitrifiers based on their ability to reduce nitrate to N2O or N2. 16S rDNA fragments amplified from the denitrifying isolates were analysed by restriction fragment length polymorphism. Isolates were grouped according to their restriction patterns, and 16S rDNA of representatives from each group were sequenced. A plant dependent enrichment of Agrobacterium-related denitrifiers has been observed resulting in a modification of the structure of the denitrifying community between planted and bulk soil. In addition, the predominant isolates in the rhizosphere soil were not able to reduce N2O while dominant isolates in the bulk soil evolve N2 as a denitrification product. PMID:15381970

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

    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 classes of students have engaged in and contributed data to science investigations. In Alaska, classes and individual students have conducted their own inquiry studies and have successfully presented their investigations and competed at science fairs and statewide high school science symposium and international conferences. Two students presented their research investigations at the GLOBE Learning Expedition in Croatia and four students presented their study at the GLOBE Arctic POPs Conference in Sweden. These students increased not only their understanding and knowledge of science but also in appreciation of people in other countries and their cultures. Friendships have also bloomed. The learning community in Alaska has expanded to include family and community members including Native elders (using OLCG), teachers, scientists and students from other countries. The following challenges remain: 1) getting funds to be able to provide GLOBE equipment and continuous support to GLOBE teachers and students throughout the year, 2) reaching teachers and students in remote areas, 3) rapid teacher turn-over rate in rural areas, 4) using inquiry-based pedagogies during GLOBE professional development workshops including the opportunity for teacher participants to conduct their own inquiries during the workshop, 5) time, school curriculum and national education requirement constraints, 6) involving school administrators, and more local scientists and community members, and 7) providing culturally relevant and responsive science education programs and life-long learning communities.

  1. Fermentative degradation of nonionic surfactants and polyethylene glycol by enrichment cultures and by pure cultures of homoacetogenic and propionate-forming bacteria.

    WAGENER, Stefan; Schink, Bernhard

    1988-01-01

    Linear alkyl ethoxylates (polyethylene glycol alkyl ethers) were fermented completely to methane and CO2 in enrichment cultures inoculated with anoxic sewage sludge. Long-chain fatty acids were released as intermediates. No degradation was found with polypropylene glycol and polypropylene glycol-containing surfactants. Two types of primary ethoxylate-degrading bacteria were isolated and characterized. Both degraded polyethylene glycols with molecular weights of 1,000 completely. Strain KoB35 ...

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

    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)

  3. Differential requirements for enriched atmospheric carbon dioxide content for intracellular growth in cell culture among selected members of the genus Rickettsia.

    Kopmans-Gargantiel, A I; Wisseman, C. L.

    1981-01-01

    In an in vitro chicken embryo cell culture system, strains of Rickettsia prowazekii, R. mooseri, and R. rickettsii, but not of R. tsutsugamushi, required an atmosphere enriched in CO2 for intracellular growth.

  4. Biodiversity within hot spring microbial mat communities: molecular monitoring of enrichment cultures

    Ward, D. M.; Santegoeds, C. M.; Nold, S. C.; Ramsing, N. B.; Ferris, M. J.; Bateson, M. M.

    1997-01-01

    We have begun to examine the basis for incongruence between hot spring microbial mat populations detected by cultivation or by 16S rRNA methods. We used denaturing gradient gel electrophoresis (DGGE) to monitor enrichments and isolates plated therefrom. At near extincting inoculum dilutions we observed Chloroflexus-like and cyanobacterial populations whose 16S rRNA sequences have been detected in the 'New Pit' Spring Chloroflexus mat and the Octopus Spring cyanobacterial mat. Cyanobacterial populations enriched from 44 to 54 degrees C and 56 to 63 degrees C samples at near habitat temperatures were similar to those previously detected in mat samples of comparable temperatures. However, a lower temperature enrichment from the higher temperature sample selected for the populations found in the lower temperature sample. Three Thermus populations detected by both DGGE and isolation exemplify even more how enrichment may bias our view of community structure. The most abundant population was adapted to the habitat temperature (50 degrees C), while populations adapted to 65 degrees C and 70 degrees C were 10(2)- and 10(4)-fold less abundant, respectively. However, enrichment at 70 degrees C favored the least abundant strain. Inoculum dilution and incubation at the habitat temperature favored the more numerically relevant populations. We enriched many other aerobic chemoorganotrophic populations at various inoculum dilutions and substrate concentrations, most of whose 16S rRNA sequences have not been detected in mats. A common feature of numerically relevant cyanobacterial, Chloroflexus-like and aerobic chemorganotrophic populations, is that they grow poorly and resist cultivation on solidified medium, suggesting plating bias, and that the medium composition and incubation conditions may not reflect the natural microenvironments these populations inhabit.

  5. Functional consortium for denitrifying sulfide removal process

    Chen, Chuan [Harbin Inst. of Technology (CN). State Key Lab. of Water Resource and Environment (SKLWRE); Harbin Inst. of Technology (China). School of Municipal and Environmental Engineering; Ren, Nanqi; Wang, Aijie [Harbin Inst. of Technology (CN). State Key Lab. of Water Resource and Environment (SKLWRE); Liu, Lihong [Harbin Inst. of Technology (China). School of Municipal and Environmental Engineering; Lee, Duu-Jong [Harbin Inst. of Technology (CN). State Key Lab. of Water Resource and Environment (SKLWRE); National Taiwan Univ., Taipei (China). Dept. of Chemical Engineering

    2010-03-15

    Denitrifying sulfide removal (DSR) process simultaneously converts sulfide, nitrate, and chemical oxygen demand from industrial wastewaters to elemental sulfur, nitrogen gas, and carbon dioxide, respectively. This investigation utilizes a dilution-to-extinction approach at 10{sup -2} to 10{sup -6} dilutions to elucidate the correlation between the composition of the microbial community and the DSR performance. In the original suspension and in 10{sup -2} dilution, the strains Stenotrophomonas sp., Thauera sp., and Azoarcus sp. are the heterotrophic denitrifiers and the strains Paracoccus sp. and Pseudomonas sp. are the sulfide-oxidizing denitrifers. The 10{sup -4} dilution is identified as the functional consortium for the present DSR system, which comprises two functional strains, Stenotrophomonas sp. strain Paracoccus sp. At 10{sup -6} dilution, all DSR performance was lost. The functions of the constituent cells in the DSR granules were discussed based on data obtained using the dilution-to-extinction approach. (orig.)

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

    Jiang, Yang; Mikova, Gizela; Kleerebezem, Robbert; van der Wielen, Luuk Am; Cuellar, Maria C

    2015-12-01

    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 (0.2 w/v% SDS) could reach 99% purity, with more than 90% recovery. The lost PHB could be mostly attributed to PHB hydrolysis during the alkaline treatment. PHB hydrolysis could be moderated by increasing the crystallinity of the PHB granules, for example, by biomass pretreatment (e.g. freezing or lyophilization) or by effective cell lysis (e.g. adjusting alkali concentration). The suitability of the purified PHB by alkaline treatment for polymer applications was evaluated by molecular weight and thermal stability. A solvent based purification method was also performed for comparison purposes. As result, PHB produced by mixed enriched cultures was found suitable for thermoplastic applications when purified by the solvent method. While the alkaline method resulted in purity, recovery yield and molecular weight comparable to values reported in literature for PHB produced by pure cultures, it was found unsuitable for thermoplastic applications. Given the potential low cost and favorable environmental impact of this method, it is expected that PHB purified by alkaline method may be suitable for other non-thermal polymer applications, and as a platform chemical. PMID:25642402

  7. The influence of fertilization on the quantitative distribution of denitrifying bacteria in the soil

    Simona Dunca; Marius Stefan; Mariana Murgoci

    2010-01-01

    Highlighting the processes by which the nitrogen compounds are reduced in the soil (i.e. denitrification) and which result in the increase of the ammonium level, is extremely important since denitrification is one of the effective mechanisms that reduce the nitrogen content in the soils rich in nitrites thus preventing soil pollution. The determination of the most probable number of denitrifying bacteria was carried out by the multiple tube technique using Allens culture medium, ...

  8. A New Protocol to Detect Multiple Foodborne Pathogens with PCR Dipstick DNA Chromatography after a Six-Hour Enrichment Culture in a Broad-Range Food Pathogen Enrichment Broth

    Masahiro Hayashi; Tatsuya Natori; Sayoko Kubota-Hayashi; Machiko Miyata; Kiyofumi Ohkusu; Keiko Kawamoto; Hisao Kurazono; Souichi Makino; Takayuki Ezaki

    2013-01-01

    A quick foodborne pathogen screening method after six-hour enrichment culture with a broad-range food pathogen enrichment broth is described. Pathogenic factors of Salmonella enterica, Shigella spp., enteroinvasive Escherichia coli, and enterohemorrhagic E. coli are amplified with a cocktail primer and rapid polymerase chain reaction (PCR), which finishes amplification in 30 min. The PCR amplicon was differentiated with a dipstick DNA chromatography assay in 5–10 min. Starting from a four- to...

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

    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…

  10. Enrichment and identification of ?9-Tetrahydrocannabinolic acid synthase from Pichia pastoris culture supernatants

    Kerstin Lange

    2015-09-01

    Full Text Available This data article refers to the report ?9-Tetrahydrocannabinolic acid synthase (THCAS production in Pichia pastoris enables chemical synthesis of cannabinoids (Lange et. al. 2015 [2]. THCAS was produced on a 2L lab scale using recombinant P. pastoris KM71 KE1. Enrichment of THCAS as a technically pure enzyme was realized using dialysis and cationic exchange chromatography. nLC-ESI-MS/MS analysis identified THCAS in different fractions obtained by cationic exchange chromatography.

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

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

    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 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. PMID:26905741

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

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

    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 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. PMID:26905741

  13. Characterization of diverse heterocyclic amine-degrading denitrifying bacteria from various environments.

    Bae, Hee-Sung; Im, Wan-Taek; Suwa, Yuichi; Lee, James M; Lee, Sung-Taik; Chang, Young-Keun

    2009-04-01

    Although, there have been many published bacterial strains aerobically degrading the heterocyclic amine compounds, only one strain to date has been reported to degrade pyrrolidine under denitrifying conditions. In this study, denitrifying bacteria degrading pyrrolidine and piperidine were isolated from diverse geological and ecological origins through selective enrichment procedures. Based on the comparative sequence results of 16S rRNA genes, 30 heterocyclic amine-degrading isolates were grouped into ten distinct phylotypes belonging to the genera Thauera, Castellaniella, Rhizobium, or Paracoccus of the phylum Proteobacteria. The representative isolates of individual phylotypes were characterized by phylogenetic, phenotypic and chemotaxonomical traits, and dissimilatory nitrite reductase gene (nirK and nirS). All isolates completely degraded pyrrolidine and piperidine under both aerobic and anaerobic conditions. The anaerobic degradations were coupled to nitrate reduction. A metabolic pathway for the anaerobic degradation of pyrrolidine was proposed on the basis of enzyme activities implicated in pyrrolidine metabolism from three isolates. The three key pyrrolidine-metabolizing enzymes pyrrolidine dehydrogenase, gamma-aminobutyrate/alpha-ketoglutarate aminotransferase, and succinic semialdehyde dehydrogenase, were induced by heterocyclic amines under denitrifying conditions. They were also induced in cells grown aerobically on heterocyclic amines, suggesting that the anaerobic degradation of pyrrolidine shares the pathway with aerobic degradation. PMID:19183960

  14. Study of selenocompounds from selenium-enriched culture of edible sprouts

    Funes Collado, Virginia; Morell García, Albert; Rubio i Rovira, Roser; López Sánchez, José Fermín

    2013-01-01

    Selenium is recognised as an essential micronutrient for humans and animals. One of the main sources of selenocompounds in the human diet is vegetables. Therefore, this study deals with the Se species present in different edible sprouts grown in Se-enriched media. We grew alfalfa, lentil and soy in a hydroponic system amended with soluble salts, containing the same proportion of Se, in the form of Se(VI) and Se(IV). Total Se in the sprouts was determined by acidic digestion in a microwave sys...

  15. Anaerobic degradation of 3-aminobenzoate by a newly isolated sulfate reducer and a methanogenic enrichment culture

    Schnell, Sylvia; Schink, Bernhard

    1992-01-01

    A new rod-shaped, gram-negative, non-sporing sulfate reducer, strain mAB1, was enriched and isolated from marine sediment samples with 3-aminobenzoate as sole electron and carbon source. Strain mAB1 degraded 3-aminobenzoate completely to CO 2 and NH 3 with stoichiometric reduction of sulfate to sulfide. Ceils contained carbon monoxide dehydrogenase, cytochromes, and sulfite reductase P582. Strain mAB1 degraded also benzoate, 4-aminobenzoate, hydroxybenzoates, and some aliphatic compounds. Bes...

  16. A new protocol to detect multiple foodborne pathogens with PCR dipstick DNA chromatography after a six-hour enrichment culture in a broad-range food pathogen enrichment broth.

    Hayashi, Masahiro; Natori, Tatsuya; Kubota-Hayashi, Sayoko; Miyata, Machiko; Ohkusu, Kiyofumi; Kawamoto, Keiko; Kurazono, Hisao; Makino, Souichi; Ezaki, Takayuki

    2013-01-01

    A quick foodborne pathogen screening method after six-hour enrichment culture with a broad-range food pathogen enrichment broth is described. Pathogenic factors of Salmonella enterica, Shigella spp., enteroinvasive Escherichia coli, and enterohemorrhagic E. coli are amplified with a cocktail primer and rapid polymerase chain reaction (PCR), which finishes amplification in 30 min. The PCR amplicon was differentiated with a dipstick DNA chromatography assay in 5-10 min. Starting from a four- to six-hour enrichment culture, this assay was finished within 45 min. Detection sensitivity of this protocol was less than 2.5 CFU/25 g for S. enterica and 3.3 CFU/25 g for enterohemorrhagic E. coli in spiked ground meat experiments. PMID:24364031

  17. The Control of Microcystis spp. Bloom by Combining Indigenous Denitrifying Bacteria From Sutami Reservoir with Fimbristylis globulosa and Vetiveria zizanoides

    Bayu Agung Prahardika

    2013-04-01

    Full Text Available The purpose of this research is to know the ability of polyculture macrophyte (Fimbristylis globulosa and Vetiveria zizanoides and the combination of both with consortium of indigenous denitrifying bacteria from Sutami reservoir that was added by Microcystis spp. or not to reduce the concentration of nitrate, dissolved phosphate and the carrying capacity of Microcystis spp. The experiment was done in a medium filled up with Sutami reservoir water enriched with 16 ppm of nitrate and 0.4 ppm of phosphate. The denitrifying bacteria used in this research were DR-14, DU-27-1, DU-30-1, DU-30-2, TA-8 and DU-27-4 isolated from Sutami reservoir. The treatments were incubated within 15 days. Microcystis spp. abundance was calculated every day, but the measurement of the concentration of nitrate and dissolved phosphate was done every six days. The results showed that both treatment and the combination of both macrophytes with a consortium of denitrifying indigenous bacteria were added or not either Microcystis able to reduce nitrate at 99% and 93-99% orthophosphoric. The combination of macrophytes with denitrifying indigenous bacterial consortium from Sutami reservoir was able to inhibit the carrying capacity of Microcystis spp. highest up to 47.87%. They could also significantly reduce the abundance of Microcystis from 107 cells/mL in earlier days of the treatment into 0.35x104 cells/mL after fifteen days of incubation.

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

    Fotidis, Ioannis; Wang, Han; Angelidaki, Irini

    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...

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

    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.

  20. Growth and nitrate reduction of Beggiatoa filaments studied in enrichment cultures

    Kamp, Anja

    nitrate, sulphide, oxygen, and pH were determined with microsensors. The more nitrate that was added to the agar, the deeper the Beggiatoa filaments glided into anoxic agar layers, suggesting that the Beggiatoa used nitrate to oxidise sulphide at depths below the depth that oxygen penetrated. In the...... oxidation. This interpretation is consistent with a fourfold-lower oxygen flux and a twofold- higher sulphide flux into the nitrate-exposed mats compared to the fluxes for controls without nitrate. Additionally, a pronounced pH maximum was observed within the Beggiatoa mat; such a pH maximum is known to......In this thesis, several aspects of the gliding, filamentous, colourless sulphur bacteria Beggiatoa were investigated. The first part of this thesis addressed the growth mode, breakage of filaments for multiplication, and movement directions of filaments of Beggiatoa. Marine Beggiatoa were enriched...

  1. Rapid and Specific Enrichment of Culturable Gram Negative Bacteria Using Non-Lethal Copper-Free Click Chemistry Coupled with Magnetic Beads Separation

    Fugier, Emilie; Dumont, Audrey; Malleron, Annie; Poquet, Enora; Mas Pons, Jordi; Baron, Aurélie; Vauzeilles, Boris; Dukan, Sam

    2015-01-01

    Currently, identification of pathogenic bacteria present at very low concentration requires a preliminary culture-based enrichment step. Many research efforts focus on the possibility to shorten this pre-enrichment step which is needed to reach the minimal number of cells that allows efficient identification. Rapid microbiological controls are a real public health issue and are required in food processing, water quality assessment or clinical pathology. Thus, the development of new methods fo...

  2. Biopolymer Production by Bacterial Enrichment Cultures Using Non-Fermented Substrates :

    Moralejo Gárate, H.

    2014-01-01

    Polyhydroxyalkanotes (PHAs) are naturally occurring polymers synthesized by a wide range of microorganisms. Their physiological role is to act as carbon and energy reserves, and their mechanical and physical properties are similar to those of petrochemical plastics. PHAs can be synthesized from renewable materials and they are biodegradable. Considering these properties, PHAs are commonly known as bioplastics. Commercial processes for the production of PHA are based on the use of pure culture...

  3. Proteins differentially expressed in human beta-cells-enriched pancreatic islet cultures and human insulinomas

    Terra, Letícia F; Teixeira, Priscila C; Wailemann, Rosangela A M; Zelanis, André; Palmisano, Giuseppe; Cunha-Neto, Edecio; Kalil, Jorge; Larsen, Martin Røssel; Labriola, Leticia; Sogayar, Mari C

    those of primary beta-cells using DIGE followed by MS. The results were validated by Western blotting. An average of 1800 spots was detected with less than 1% exhibiting differential abundance. Proteins more abundant in human islets, such as Caldesmon, are involved in the regulation of cell...... source of cultured human beta-cells for beta-cell research, along with the development of new therapeutic strategies for detection, characterization and treatment of insulinomas....

  4. Biopolymer Production by Bacterial Enrichment Cultures Using Non-Fermented Substrates

    Moralejo Gárate, H.

    2014-01-01

    Polyhydroxyalkanotes (PHAs) are naturally occurring polymers synthesized by a wide range of microorganisms. Their physiological role is to act as carbon and energy reserves, and their mechanical and physical properties are similar to those of petrochemical plastics. PHAs can be synthesized from renewable materials and they are biodegradable. Considering these properties, PHAs are commonly known as bioplastics. Commercial processes for the production of PHA are based on the use of pure culture...

  5. Denitrification by intact soybean nodules in relation to natural 15N enrichment of nodules

    The natural 15N abundance of nodules of soybeans (Glycine max (L.) Merrill) which are actively fixing N2 is considerably higher than other tissues. To investigate the question of whether isotopic fractionation associated with denitrification by bacteroids causes this 15N enrichment, we inoculated soybeans with two strains of Rhizobium japonicum. Free-living cultures of one of these (strain USDA 33) were unable to denitrify or respire NO3-, while free-living cultures of the second (strain USDA 138) were capable of denitrification. USDA 138 formed nodules which fixed N2 very efficiently. The N of these nodules was enriched in 15N and the nodules reduced a substantial amount of NO3- to NO2- and N2O. Nodules infected with USDA 33 fixed about half as much N2 as those infected with USDA 138. The former nodules were enriched in 15N (although less so than nodules infected with USDA 138), despite the fact that the nodules formed by USDA 33 did not reduce NO3-. Clearly denitrification could not have been the cause of 15N enrichment of nodules infected with strain USDA 33. Alternative causes of 15N enrichment of soybean nodules and their possible metabolic significance are discussed

  6. Production of polyhydroxyalkanoates from fermented sugar cane molasses by a mixed culture enriched in glycogen accumulating organisms.

    Bengtsson, Simon; Pisco, Ana R; Reis, Maria A M; Lemos, Paulo C

    2010-02-01

    Batch production of polyhydroxyalkanoates (PHAs) under aerobic conditions by an open mixed culture enriched in glycogen accumulating organisms (GAOs) with fermented sugar cane molasses as substrate was studied. The produced polymers contained five types of monomers, namely 3-hydroxybutyrate (3HB), 3-hydroxyvalerate (3HV), 3-hydroxy-2-methylbutyrate (3H2MB), 3-hydroxy-2-methylvalerate (3H2MV) and the medium chain length monomer 3-hydroxyhexanoate (3HHx). With fermented molasses as substrate, PHA was produced under concurrent consumption of stored glycogen with yields of 0.47-0.66 C-mol PHA per C-mol of total carbon substrate and with rates up to 0.65 C-mol/C-molX h. In order to investigate the role of glycogen during aerobic PHA accumulation in GAOs, synthetic single volatile fatty acids (VFAs) were used as substrates and it was found that the fate of glycogen was dependent on the type of VFA being consumed. Aerobic PHA accumulation occurred under concurrent glycogen consumption with acetate as substrate and under minor concurrent glycogen production with propionate as substrate. With butyrate and valerate as substrates, PHA accumulation occurred with the glycogen pool unaffected. The composition of the PHA was dependent on the VFA composition of the fermented molasses and was 56-70 mol-% 3HB, 13-43 mol-% 3HV, 1-23 mol-% 3HHx and 0-2 mol-% 3H2MB and 3H2MV. The high polymer yields and production rates suggest that enrichment of GAOs can be a fruitful strategy for mixed culture production of PHA from waste substrates. PMID:19958801

  7. Pilot-scale chitin extraction from shrimp shell waste by deproteination and decalcification with bacterial enrichment cultures.

    Bajaj, Mini; Freiberg, Andrea; Winter, Josef; Xu, Youmei; Gallert, Claudia

    2015-11-01

    Extraction of chitin from mechanically pre-purified shrimp shells can be achieved by successive NaOH/HCl treatment, protease/HCl treatment or by environmentally friendly fermentation with proteolytic/lactic acid bacteria (LAB). For the last mentioned alternative, scale-up of shrimp shell chitin purification was investigated in 0.25 L (F1), 10 L (F2), and 300 L (F3) fermenters using an anaerobic, chitinase-deficient, proteolytic enrichment culture from ground meat for deproteination and a mixed culture of LAB from bio-yoghurt for decalcification. Protein removal in F1, F2, and F3 proceeded in parallel within 40 h at an efficiency of 89-91 %. Between 85 and 90 % of the calcit was removed from the shells by LAB in another 40 h in F1, F2, and F3. After deproteination of shrimp shells in F3, spent fermentation liquor was re-used for a next batch of 30-kg shrimp shells in F4 (300 L) which eliminated 85.5 % protein. The purity of the resulting chitin was comparable in F1, F2, F3, and F4. Viscosities of chitosan, obtained after chitin deacetylation and of chitin, prepared biologically or chemically in the laboratory, were much higher than those of commercially available chitin and chitosan. PMID:26227412

  8. Enrichment of CD8+ cells from melanoma tumor-infiltrating lymphocyte cultures reveals tumor reactivity for use in adoptive cell therapy.

    Prieto, Peter A; Durflinger, Katherine H; Wunderlich, John R; Rosenberg, Steven A; Dudley, Mark E

    2010-06-01

    Adoptive cell therapy (ACT) using tumor-infiltrating lymphocytes (TIL) for metastatic melanoma has shown objective response rates as high as 72%. The successful application of this therapy requires the selection of unique tumor-reactive lymphocyte cultures for each patient. This is a technically and logistically difficult undertaking, and patients who do not have tumor-reactive TIL are not considered eligible for treatment. To simplify the methods of TIL generation and extend TIL-based immunotherapy to additional patients, methods were developed to use unselected, minimally cultured ("young") TIL. Young TIL cultures contain a variable number of CD8(+), CD4(+), and CD3(-)CD56(+) natural killer cells. In this study we retrospectively investigated a role for these subsets in the clinical outcome of patients treated with TIL derived from selected microcultures. This analysis demonstrated a suggestive but nonsignificant association between the number of CD8(+) cells administered and tumor regression. We therefore investigated the feasibility of selecting CD8(+) cells from young TIL cultures for ACT therapy. The available methods for clinical scale CD8(+) enrichment proved inadequate for TIL, so an optimized CD8(+) enrichment method was developed and is reported here. We observed that CD8 (+)enrichment of some TIL cultures revealed in vitro tumor recognition that was not evident in bulk culture, and an improved in vitro recognition of tumor in other TIL cultures. In addition, the enriched CD8(+) young TIL expanded more reliably and predictably in rapid expansions than the bulk TIL. Thus, optimized CD8(+) selection combines the benefits of antigen-selected TIL and young TIL for generating lymphocyte cultures for ACT, and should be evaluated in cell transfer therapy protocols. PMID:20463593

  9. New developments in the mass culture and nutritional enrichment of the rotifer Brachionus plicatilis using artificial diets

    Komis, A.; Naessens, E.; Van Ballaer, E.; Van Sprang, P; Léger, P.; Sorgeloos, P

    1989-01-01

    A research program was carried out to improve and facilitate rotifer (Brachionus plicatilis) culturing as well as to assure their optimal nutritional quality as a prey, concentrating on the development of a dry and complete rotifer diet that does not require the use of algae and does result in a HUFA rich Brachionus. The present poster provides results dealing with culture aspects and rotifer nutritional quality. Different diet compositions based on single cell proteins enriched with various ...

  10. Distribution of Microorganisms in Deep-Sea Hydrothermal Vent Chimneys Investigated by Whole-Cell Hybridization and Enrichment Culture of Thermophilic Subpopulations

    Harmsen, H; Prieur, D; Jeanthon, C

    1997-01-01

    The microbial community structure of hydrothermal vent chimneys was evaluated by the combined use of enrichment cultures and whole-cell hybridizations with fluorescently labeled 16S rRNA-based oligonucleotide probes. Chimneys were collected during the Microsmoke cruise on the Mid-Atlantic Ridge and were subsampled on board and stored under reduced conditions or fixed. For estimation of culturable thermophiles, selective media were inoculated by dilution series of the samples and incubated at ...

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

    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 53.92g mL(-1), 265.6 and 1805.2g mL(-1) and 1.85 and 16.12g 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, however, was degraded up to T4 treatments and was persistent in the T5 treatment. Probably, accumulation of this metabolite inhibited atrazine/cyanuric acid degradation by the enrichment culture in undiluted wastewater. PMID:26479154

  12. Comparative and multilaboratory studies of two immunodiffusion method enrichment protocols and the AOAC/Bacteriological Analytical Manual culture method for detection of Salmonella in all foods.

    Feldsine, P T; Falbo-Nelson, M T; Hustead, D L; Flowers, R S; Flowers, M J

    1995-01-01

    The single enrichment immunodiffusion (1-step), the preenrichment and selective enrichment immunodiffusion (2-step), and the AOAC/Bacteriological Analytical Manual culture methods for Salmonella were evaluated for equivalence in 2 separate studies, a comparative evaluation and a multilaboratory dilution study. In the comparative study, all 3 methods were performed on 10 food types. For 550 samples, analyses resulted in 99.3 and 99.6% agreement between the culture method and the 1-step and 2-step methods, respectively. False negative rates were 0.9 and 0.3% for 1-step and culture, and 0.0% and 0.6% for the 2-step and culture, respectively. Subsequently, 6 food types were included in a multilaboratory dilution-to-extinction study. A sequential dilution series of Salmonella in foods was analyzed by the 3 methods to determine their lower limits of detection for Salmonella. A total of 1185 samples analyzed resulted in 98.9% agreement between 1-step and culture, and 99.7% agreement between 2-step and culture. False negative rates were 1.8 and 0.1% for 1-step and culture, and 0.4 and 0.1% for 2-step and culture, respectively. During these evaluations, 1735 samples and controls representing 10 different naturally contaminated and inoculated foods were tested. The data indicate statistical equivalence of all 3 methods when analyzing all food types. PMID:7580340

  13. Cultivation and irradiation of human fibroblasts in a medium enriched with platelet lysate for obtaining feeder layer in epidermal cell culture

    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)

  14. Rapid and Specific Enrichment of Culturable Gram Negative Bacteria Using Non-Lethal Copper-Free Click Chemistry Coupled with Magnetic Beads Separation.

    Fugier, Emilie; Dumont, Audrey; Malleron, Annie; Poquet, Enora; Mas Pons, Jordi; Baron, Aurélie; Vauzeilles, Boris; Dukan, Sam

    2015-01-01

    Currently, identification of pathogenic bacteria present at very low concentration requires a preliminary culture-based enrichment step. Many research efforts focus on the possibility to shorten this pre-enrichment step which is needed to reach the minimal number of cells that allows efficient identification. Rapid microbiological controls are a real public health issue and are required in food processing, water quality assessment or clinical pathology. Thus, the development of new methods for faster detection and isolation of pathogenic culturable bacteria is necessary. Here we describe a specific enrichment technique for culturable Gram negative bacteria, based on non-lethal click chemistry and the use of magnetic beads that allows fast detection and isolation. The assimilation and incorporation of an analog of Kdo, an essential component of lipopolysaccharides, possessing a bio-orthogonal azido function (Kdo-N3), allow functionalization of almost all Gram negative bacteria at the membrane level. Detection can be realized through strain-promoted azide-cyclooctyne cycloaddition, an example of click chemistry, which interestingly does not affect bacterial growth. Using E. coli as an example of Gram negative bacterium, we demonstrate the excellent specificity of the technique to detect culturable E. coli among bacterial mixtures also containing either dead E. coli, or live B. subtilis (as a model of microorganism not containing Kdo). Finally, in order to specifically isolate and concentrate culturable E. coli cells, we performed separation using magnetic beads in combination with click chemistry. This work highlights the efficiency of our technique to rapidly enrich and concentrate culturable Gram negative bacteria among other microorganisms that do not possess Kdo within their cell envelope. PMID:26061695

  15. Drinking Water Denitrification using Autotrophic Denitrifying Bacteria in a Fluidized Bed Bioreactor 

    Abdolmotaleb Seid-mohammadi

    2013-02-01

    Full Text Available Background and Objectives: Contamination of drinking water sources with nitrate may cause adverse effects on human health. Due to operational and maintenance problems of physicochemical nitrate removal processes, using biological denitrification processes have been performed. The aim of this study is to evaluate nitrate removal efficiency from drinking water using autotrophic denitrifying bacteria immobilized on sulfur impregnated activated carbon in a fluidized bed bioreactor. Materials and Methods: After impregnating activated carbon by sulfur as a microorganism carriers and enrichment and inoculation of denitrifying bacteria, a laboratory-scale fluidized bed bioreactor was operated. Nitrate removal efficiency, nitrite, turbidity, hardness and TOC in the effluent were examined during the whole experiment under various conditions including constant influent nitrate concentration as 90 mg NO3--N/l corresponding to different HRT ranging from 5.53 to 1.5 hr. Results: We found that  the denitrification rates was depended on the hydraulic retention time and the nitrate removal efficiency was up to 98%  and nitrite concentration was lower than 1mg/l at optimum HRT=2.4 hr respectively. Moreover, there was no difference in hardness between influent and effluent due to supplying sodium bicarbonate as carbon source for denitrifying bacteria.  However pH, TOC, hardness, and turbidity of the effluent met the W.H.O guidelines for drinking water.  Conclusion: This study demonstrated that an innovative carrier as sulfur impregnated activated carbon could be used as both the biofilm carrier and energy source for treating nitrate contaminated drinking water in the lab-scale fluidized bed bioreactor.

  16. Characterization of a murine neuron-enriched model of primary trigeminal ganglia cultures to study the interferon- β antiviral effect against Herpes Simplex Virus type 1

    Ana Maria Low-Calle

    2014-06-01

    Full Text Available Objective: To characterize a neuron-enriched primary TG culture and evaluate interferon- β expression and activity after HSV-1 infection. Materials and methods: The percentage of neurons present in cultures was assessed by neurofilament immunocytochemistry. Cultures were treated with interferon- β and infected with HSV-1, then viral antigen positive cells were counted and interferon- βexpression was assessed by quantitative PCR. Results: The culture contained 15% neurons and 85% non-neuronal cells. A cytopathic effect was observed, associated with high viral spread (72.9% neurons and 48.3% non-neuronal cells were positive for viral antigen. Interferon- β treatment impaired the cytopathic effect and decreased the infected neurons to 16.7% and infected non-neuronal cells to 7.8%. Viral infection at 6 h postinfection significantly increased the interferon- β transcripts by 18.2 fold, while at 18 h postinfection Interferon pre-treatment in infected cultures increased interferon- β transcription by 3.7 fold. Discussion: This culture model contained 15% neurons, which is 10 times higher compared to other reported cultures, and non-neuronal cells comprised 85% of cells in this culture. All types of cells were found to be infected, which is similar to that reported during acute infections in vivo . Additionally, interferon- βdecreased the infected cells, avoiding the cytopathic effect, which is similar to that reported in swine TG cultures. Conclusions: A neuron-enriched primary TG model was characterized. Interferon- β treatment protected cells from cytopathic effects and viral spread, while viral infection up-regulated interferon- β expression. This result means that interferon- β exerts an important antiviral effect against HSV-1 in these cultures.

  17. Denitrifying capability and community dynamics of glycogen accumulating organisms during sludge granulation in an anaerobic-aerobic sequencing batch reactor

    Bin, Zhang; Bin, Xue; Zhigang, Qiu; Zhiqiang, Chen; Junwen, Li; Taishi, Gong; Wenci, Zou; Jingfeng, Wang

    2015-08-01

    Denitrifying capability of glycogen accumulating organisms (GAOs) has received great attention in environmental science and microbial ecology. Combining this ability with granule processes would be an interesting attempt. Here, a laboratory-scale sequencing batch reactor (SBR) was operated to enrich GAOs and enable sludge granulation. The results showed that the GAO granules were cultivated successfully and the granules had denitrifying capability. The batch experiments demonstrated that all NO3--N could be removed or reduced, some amount of NO2--N were accumulated in the reactor, and N2 was the main gaseous product. SEM analysis suggested that the granules were tightly packed with a large amount of tetrad-forming organisms (TFOs); filamentous bacteria served as the supporting structures for the granules. The microbial community structure of GAO granules was differed substantially from the inoculant conventional activated sludge. Most of the bacteria in the seed sludge grouped with members of Proteobacterium. FISH analysis confirmed that GAOs were the predominant members in the granules and were distributed evenly throughout the granular space. In contrast, PAOs were severely inhibited. Overall, cultivation of the GAO granules and utilizing their denitrifying capability can provide us with a new approach of nitrogen removal and saving more energy.

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

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

    2012-01-01

    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 phototrophic and mixotrophic protists from Lake Bonney. Sampling depths in the water column were chosen based on the location of primary production maxima and protist phylogenetic diversity (4, 11), as well as variability in major abiotic factors affecting protist trophic modes: shallow sampling depths are limited for major nutrients, while deeper sampling depths are limited by light availability. In addition, lake water samples were supplemented with multiple types of growth media to promote the growth of a variety of phototrophic organisms. RubisCO catalyzes the rate limiting step in the Calvin Benson Bassham (CBB) cycle, the major pathway by which autotrophic organisms fix inorganic carbon and provide organic carbon for higher trophic levels in aquatic and terrestrial food webs (12). In this study, we applied a radioisotope assay modified for filtered samples (13) to monitor maximum carboxylase activity as a proxy for carbon fixation potential and metabolic versatility in the Lake Bonney enrichment cultures. PMID:22546995

  19. Comparative metagenomics of three Dehalococcoides-containing enrichment cultures: the role of the non-dechlorinating community

    Hug Laura A

    2012-07-01

    Full Text Available Abstract Background The Dehalococcoides are strictly anaerobic bacteria that gain metabolic energy via the oxidation of H2 coupled to the reduction of halogenated organic compounds. Dehalococcoides spp. grow best in mixed microbial consortia, relying on non-dechlorinating members to provide essential nutrients and maintain anaerobic conditions. A metagenome sequence was generated for the dechlorinating mixed microbial consortium KB-1. A comparative metagenomic study utilizing two additional metagenome sequences for Dehalococcoides-containing dechlorinating microbial consortia was undertaken to identify common features that are provided by the non-dechlorinating community and are potentially essential to Dehalococcoides growth. Results The KB-1 metagenome contained eighteen novel homologs to reductive dehalogenase genes. The metagenomes obtained from the three consortia were automatically annotated using the MG-RAST server, from which statistically significant differences in community composition and metabolic profiles were determined. Examination of specific metabolic pathways, including corrinoid synthesis, methionine synthesis, oxygen scavenging, and electron-donor metabolism identified the Firmicutes, methanogenic Archaea, and the ?-Proteobacteria as key organisms encoding these pathways, and thus potentially producing metabolites required for Dehalococcoides growth. Conclusions Comparative metagenomics of the three Dehalococcoides-containing consortia identified that similarities across the three consortia are more apparent at the functional level than at the taxonomic level, indicating the non-dechlorinating organisms identities can vary provided they fill the same niche within a consortium. Functional redundancy was identified in each metabolic pathway of interest, with key processes encoded by multiple taxonomic groups. This redundancy likely contributes to the robust growth and dechlorination rates in dechlorinating enrichment cultures.

  20. Accumulation of nitrite in denitrifying barriers when phosphate is limiting

    Hunter, W. J.

    2003-10-01

    Permeable in situ denitrifying barriers can remove nitrate from groundwater. Barriers may be constructed by filling an excavated area with a porous mixture of sand, fine gravel, and substrate or by the injection of a nonaqueous phase substrate into an aquifer. The substrate stimulates the development of a denitrifying microbial community by providing an electron donor. The objective of this study was to determine the ability of denitrifying barriers to function under low-phosphate conditions. Sand columns injected with a soybean oil emulsion were used as laboratory models of denitrifying barriers. When a natural groundwater containing 17 mg l -1 nitrate-N and 0.009 mg l -1 phosphate-P was pumped through the columns, only a small amount of nitrate was removed from the water and, in some effluent fractions, 52% to 88% of the influent nitrate had converted to nitrite. Nitrite also accumulated when the phosphate concentration of the groundwater was increased to 0.040 or 0.080 mg l -1 phosphate-P. Only when a 0.160 mg l -1 phosphate-P supplement was added to the groundwater was there a loss of nitrate without a large accumulation of nitrite. The addition of solid calcium phosphate or rock phosphate to the sand columns was found to provide adequate phosphate for denitrification in short-term studies. These studies point out the need to ensure that adequate phosphate is present in denitrifying barriers especially when such barriers are used beneath phosphate-binding soils.

  1. Inhibitory Effect of Gamma-Irradiated Chitosan on the Growth of Denitrifiers

    Javier Vilcáez

    2009-01-01

    Full Text Available In order to find an environmentally benign substitute to hazardous inhibitory agents, the inhibitory effect of γ-irradiated chitosans against a mixed culture of denitrifying bacteria was experimentally evaluated. Unlike other studies using pure aerobic cultures, the observed effect was not a complete inhibition but a transient inhibition reflected by prolonged lag phases and reduced growth rates. Raw chitosan under acid conditions (pH 6.3 exerted the strongest inhibition followed by the 100 kGy and 500 kGy irradiated chitosans, respectively. Therefore, because the molecular weight of chitosan decreases with the degree of γ-irradiation, the inhibitory properties of chitosan due to its high molecular weight were more relevant than the inhibitory properties gained due to the modification of the surface charge and/or chemical structure by γ-irradiation. High dosage of γ-irradiated appeared to increase the growth of mixed denitrifying bacteria in acid pH media. However, in neutral pH media, high dosage of γ-irradiation appeared to enhance the inhibitory effect of chitosan.

  2. Degradation of Polycyclic Aromatic Hydrocarbons at Low Temperature under Aerobic and Nitrate-Reducing Conditions in Enrichment Cultures from Northern Soils

    Eriksson, Mikael; Sodersten, Erik; Yu, Zhongtang; Dalhammar, Gunnel; Mohn, William W

    2003-01-01

    The potential for biodegradation of polycyclic aromatic hydrocarbons (PAHs) at low temperature and under anaerobic conditions is not well understood, but such biodegradation would be very useful for remediation of polluted sites. Biodegradation of a mixture of 11 different PAHs with two to five aromatic rings, each at a concentration of 10 μg/ml, was studied in enrichment cultures inoculated with samples of four northern soils. Under aerobic conditions, low temperature severely limited PAH bi...

  3. Are Isotopologue Signatures of N2O from Bacterial Denitrifiers Indicative of NOR Type?

    Well, R.; Braker, G.; Giesemann, A.; Flessa, H.

    2010-12-01

    Nitrous oxide (N2O) fluxes from soils result from its production by nitrification and denitrification and reduction during denitrification. The structure of the denitrifying microbial community contributes to the control of net N2O fluxes. Although molecular techniques are promising for identifying the active community of N2O producers, there are few data until now because methods to explore gene expression of N2O production are laborious and disregard regulation of activity at the enzyme level. The isotopologue signatures of N2O including δ18O, average δ15N (δ15Nbulk) and 15N site preference (SP = difference in δ15N between the central and peripheral N positions of the asymmetric N2O molecule) have been used to estimate the contribution of partial processes to net N2O fluxes to the atmosphere. However, the use of this approach to study N2O dynamics in soils requires knowledge of isotopic signatures of N2O precursors and isotopologue fractionation factors (ɛ) of all processes of N2O production and consumption. In contrast to δ18O and δ15Nbulk, SP is independent of precursor signatures and hence is a promising parameter here. It is assumed that SP of produced N2O is almost exclusively controlled by the enzymatic isotope effects of NO reductases (NOR). These enzymes are known to be structurally different between certain classes of N2O producers with each class causing different isotope effects (Schmidt et al., 2004). The NH2OH-to-N2O step of nitrifiers and the NO3-to-N2O step of fungal denitrifiers are associated with large site-specific 15N effects with SP of 33 to 37 ‰ (Sutka et al., 2006, 2008) while the few tested species of gram-negative bacterial denitrifiers (cNOR group) exhibited low SP of -5 to 0‰ (Sutka et al., 2006; Toyoda et al., 2005). The aim of our study was to determine site-specific fractionation factors of the NO3-to-N2O step (ɛSP) for several species of denitrifiers representing each of the known NOR-types of bacteria, i.e. cNOR, qNOR and qCuANOR. Pure cultures where incubated at 25°C under denitrifying conditions (anaerobic in the presence of 5 mM NO3-) in their preferred media for 2 weeks. Acetylene (10%) was added to the headspace to inhibit reduction of N2O. Gas samples where collected from incubation vessels and analysed for δ18O, δ15Nbulk and SP using isotope ratio mass spectrometry. ɛSP of the qNOR-type ranged between -9.7 and -18 ‰ and of the cNOR-type between -1.7 to -10.8 ‰. Our results suggest that SP of N2O produced by soil bacteria may be more negative than previously estimated since the ɛSP range of the tested cNOR-type was larger compared to earlier studies and the qNOR type tended to be even more negative. This needs to be taken into account when using isotopologue signatures of N2O to estimate other processes such as fungal denitrification, nitrification and N2O reduction by denitrifiers. References Schmidt, H.-L. et al. 2004, Rapid Comm. Mass Spectrom. 18, 2036-2040. Sutka, R.L et al. 2006, Appl. Environ. Microbiol. 72, 638-644. Sutka, R. L et al. 2008, Rapid Commun. Mass Spectrom. 22, 3989-3996. Toyoda, S. et al. 2005, Soil Biol. Biochem. 37, 1535-1545.

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

    Highlights: ► We started up microbial fuel cell (MFC) using enriched sulfate-reducing mixed culture. ► Sulfate-reducing bacteria and anode-respiring bacteria were enriched in anodic biofilms. ► The MFC effectively remove sulfate to elementary sulfur in the presence of lactate. ► 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.

  5. Isotope effects and O-exchange with water during N2O production by denitrifying fungi

    Rohe, Lena; Braker, Gesche; Well, Reinhard; Giesemann, Anette; Anderson, Traute-Heidi; Wrage-Mönnig, Nicole; Flessa, Heinz

    2013-04-01

    N2O from soil denitrification originates from bacteria and - to an unknown extent - also from fungi. In pure culture studies, bacterial and fungal denitrification showed differences in isotopomer ratios of N2O. Isotopomer ratios of N2O can be expressed as 15N site preference (SP), i.e. the difference between δ15N of the central and terminal N-position of the asymmetric N2O molecule and might be suitable to distinguish between bacterial and fungal N2O under denitrifying soil conditions. Oxygen exchange takes place between H2O and intermediates during the denitrification process. Several studies showed that O-exchange of bacterial denitrifiers varies, but information about fungal O-exchanges is lacking. The objectives of the study presented are i) to verify SP of fungal N2O reported for two strains from pure culture studies and ii) to analyze the oxygen exchange between intermediates and H2O during denitrification in fungal pure cultures. Six different fungal pure cultures known to be capable of denitrification were incubated under anaerobic conditions, either with nitrite or nitrate. Gas samples were analysed for concentration and isotopic signatures (SP, average δ15N, δ18O) of N2O. To investigate the oxygen exchange, both treatments were also established in a tracer experiment where 18O-labelled water was used in the medium. The fungal strains evaluated so far indicate similar SP as reported previously. Concerning the amount of N2O produced, the nitrite led to a higher yield than nitrate. O-exchange with 18O-labelled water during fungal denitrification was visible. Using nitrite, the exchange ranged amounted from 39% to full exchange while nitrate led to exchange rates between 13% and full exchange. Detailed results will be presented.

  6. Hydraulic flow characteristics of agricultural residues for denitrifying bioreactor media

    Denitrifying bioreactors are a promising technology to mitigate agricultural subsurface drainage nitrate-nitrogen losses, a critical water quality goal for the Upper Mississippi River Basin. This study was conducted to evaluate the hydraulic properties of agricultural residues that are potential bio...

  7. IDENTIFICATION AND ECOPHYSIOLOGY OF ACTIVE DENITRIFIERS IN ACTIVATED SLUDGE

    Hansen, Aviaja Anna; Le-Quy, Vang; Nielsen, Kåre Lehmann; Nielsen, Jeppe Lund

    reactor studies. To obtain better identification of active denitrifying communities in full-scale wastewater treatment plants (WWTPs) we applied DNA-SIP with 13C-labelled substrates, and RT-PCR of expressed denitrification genes (nirS, nirK and nosZ) upon various substrate-inductions. To come around the...

  8. N-acyl homoserine lactone-degrading microbial enrichment cultures isolated from Penaeus vannamei shrimp gut and their probiotic properties in Brachionus plicatilis cultures

    Tinh, N.T.N.; Asanka Gunasekara, R.A.Y.S.; Boon, N.; Dierckens, K.; Sorgeloos, P.; Bossier, P

    2007-01-01

    Three bacterial enrichment cultures (ECs) were isolated from the digestive tract of Pacific white shrimp Penaeus vannamei, by growing the shrimp microbial communities in a mixture of N-acyl homoserine lactone (AHL) molecules. The ECs, characterized by denaturing gradient gel electrophoresis analysis and subsequent rRNA sequencing, degraded AHL molecules in the degradation assays. Apparently, the resting cells of the ECs also degraded one of the three types of quorum-sensing signal molecules p...

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

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

    2008-01-01

    Side population (SP) cells are highly able to exclude the Hoechst 33342 dye through membrane transporters, a feature associated with cell immaturity and therefore proposed as a marker of stem cells. Herein we demonstrate that the adipose tissue derived stromal vascular fraction (SVF) contains a...... 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...... novel population of non-haematopoietic "side population" (SPCD45(-)) cells. Simultaneous qRT-PCR of 64 genes revealed that the freshly isolated SPCD45(-) was highly enriched for cells expressing genes related to stem cells, the Notch pathway, and early vascular precursors. Notably, the expression of...

  10. Clustering and nonlinear regression analysis applied to the in vitro growing of Leucoagaricus gongylophorus (Singer Möller in a culture environment enriched with different botanical extracts

    Marcelo Dias de Souza

    2011-12-01

    Full Text Available The study aimed to evaluate the in vitro growing of Leucoagaricus gongylophorus in a culture environment enriched with different extracts from botanical species, via multivariate statistical analysis of data clustering, besides identifying the fungus behavior through nonlinear regression equations. The fungus was inoculated into a BDA medium enriched with eight botanical extracts, along with two controls (with and without the addition of 20mL distilled water, in Petri plates. The plates were kept in a climatized chamber at a 25±1ºC temperature in the dark, during a 42 days period and the fungus growing was evaluated at weekly intervals. It was proved that the clustering analysis and the Chapman-Richards model are effi cient to verify the similarity of different botanical extracts in the fungus development, as well as to estimate and describe the growing of L. gongylophorus.

  11. Trace Gas Emission from in-Situ Denitrifying Bioreactors

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

    2014-12-01

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

  12. Comparative Analysis of 16S rRNA and amoA Genes from Archaea Selected with Organic and Inorganic Amendments in Enrichment Culture

    Xu, Mouzhong; Schnorr, Jon; Keibler, Brandon; Simon, Holly M.

    2012-01-01

    We took advantage of a plant-root enrichment culture system to characterize mesophilic soil archaea selected through the use of organic and inorganic amendments. Comparative analysis of 16S rRNA and amoA genes indicated that specific archaeal clades were selected under different conditions. Three amoA sequence clades were identified, while for a fourth group, identified by 16S rRNA gene analysis alone and referred to as the “root” clade, we detected no corresponding amoA gene. The amoA-contai...

  13. Alternative anaerobic enrichments to the bacteriological analytical manual culture method for isolation of Shigella sonnei from selected types of fresh produce.

    Jacobson, Andrew P; Thunberg, Richard L; Johnson, Mildred L; Hammack, Thomas S; Andrews, Wallace H

    2004-01-01

    Alternative methods of reducing oxygen during anaerobic enrichment in the Bacteriological Analytical Manual (BAM) Shigella culture method were evaluated and compared to the current and less practical GasPak method. The alternative anaerobic methods included the use of reducing agents in Shigella broth and reducing culture container headspace volume to minimize atmospheric effects on oxygen concentration in Shigella broth during enrichment. The reducing agents evaluated were sodium thioglycollate, L-cystine, L-cysteine, titanium(III) citrate, and dithiothreitol, each at concentrations of 0.1, 0.05, and 0.01%. The use of Oxyrase for Broth with the enrichment medium (Shigella broth) was evaluated at concentrations of 10, 20 and 30 microL/mL. Recoveries of chill- and freeze-stressed S. sonnei strains 357 and 20143 were determined with each anaerobic method, including the GasPak method, using inoculation levels ranging from 10(0)to 10(3) cells. For each anaerobic method, strain, inoculation level, and stress type, 5 replicate enrichments were evaluated by streaking to MacConkey agar for isolation. The numbers of cultures with each method from which S. sonnei was isolated were used to compare the alternative anaerobic methods to the GasPak method. The alternative anaerobic method with which chill- and freeze-stressed S. sonnei strains 357 and 20143 were isolated most consistently was the use of Oxyrase for Broth in Shigella broth at a concentration of 20 microL/mL. This method was compared to the GasPak anaerobic method in evaluations on the recovery of S. sonnei strains 357 and 20143 from artificially contaminated test portions of parsley, cilantro, green onions, strawberries, carrots, and celery. A third anaerobic method included the use of 0.5 cm mineral oil overlay on cultures containing Oxyrase for Broth at concentrations of 20 microL/mL. Recovery rates of strain 357 were significantly greater (p parsley, cilantro, and celery. When Oxyrase for Broth was used with Shigella broth, strain 357 was isolated at higher rates from all produce types, except cilantro, when 0.5 cm mineral oil overlay was applied to enrichment cultures. The use of mineral oil overlay with Oxyrase for Broth also improved recovery of strain 20143 from test portions of all produce types except green onion and strawberries. These differences were significant (p parsley, carrots, and cilantro (1 of 2 evaluations). No statistically significant differences (p > 0.05) between the GasPak and Oxyrase for Broth anaerobic methods occurred when mineral oil overlay was used with Oxyrase for Broth. The use of Oxyrase for Broth with a 0.5 cm mineral oil overlay is a practical alternative for anaerobic enrichment with the BAM method in the analysis of some produce types. Differences in recovery among the different produce types and methods occurred between S. sonnei strains 357 and 20143, emphasizing the need for additional S. sonnei strains in future evaluations. PMID:15493668

  14. Enhanced nitrate and phosphate removal in a denitrifying bioreactor with biochar.

    Bock, Emily; Smith, Nick; Rogers, Mark; Coleman, Brady; Reiter, Mark; Benham, Brian; Easton, Zachary M

    2015-03-01

    Denitrifying bioreactors (DNBRs) are an emerging technology used to remove nitrate-nitrogen (NO) from enriched waters by supporting denitrifying microorganisms with organic carbon in an anaerobic environment. Field-scale investigations have established successful removal of NO from agricultural drainage, but the potential for DNBRs to remediate excess phosphorus (P) exported from agricultural systems has not been addressed. We hypothesized that biochar addition to traditional woodchip DNBRs would enhance NO and P removal and reduce nitrous oxide (NO) emissions based on previous research demonstrating reduced leaching of NO and P and lower greenhouse gas production associated with biochar amendment of agricultural soils. Nine laboratory-scale DNBRs, a woodchip control, and eight different woodchip-biochar treatments were used to test the effect of biochar on nutrient removal. The biochar treatments constituted a full factorial design of three factors (biochar source material [feedstock], particle size, and application rate), each with two levels. Statistical analysis by repeated measures ANOVA showed a significant effect of biochar, time, and their interaction on NO and dissolved P removal. Average P removal of 65% was observed in the biochar treatments by 18 h, after which the concentrations remained stable, compared with an 8% increase in the control after 72 h. Biochar addition resulted in average NO removal of 86% after 18 h and 97% after 72 h, compared with only 13% at 18 h and 75% at 72 h in the control. Biochar addition also resulted in significantly lower NO production. These results suggest that biochar can reduce the design residence time by enhancing nutrient removal rates. PMID:26023979

  15. nirS-containing denitrifier communities in the water column and sediment of the Baltic Sea

    Falk, S.; Hannig, M.; Gliesche, C.; Wardenga, R.; Köster, M. (Miriam); Jürgens, K.; Braker, G.

    2007-01-01

    The aim of this study was to compare structural differences in the nirS-type denitrifying microbial communities along the environmental gradients observed in the water column and coastal sediments of the Baltic Sea. To link community structure and environmental gradients, denitrifier communities were analyzed by terminal restriction fragment length polymorphism (T-RFLP) based on nirS as a functional marker gene for denitrification. nirS-type denitrifier community composition...

  16. Complete genome sequence of Methylophilus sp. TWE2 isolated from methane oxidation enrichment culture of tap-water.

    Xia, Fei; Zou, Bin; Shen, Cong; Zhu, Ting; Gao, Xin-Hua; Quan, Zhe-Xue

    2015-10-10

    The non-methane-utilizing methylotroph, Methylophilus sp. TWE2, was isolated from tap-water during the enrichment of methanotrophs with methane. The complete genome sequence of strain TWE2 showed that this bacterium may convert methanol to formaldehyde via catalysis of methanol dehydrogenase (MDH), after which formaldehyde would be assimilated to biomass through the ribulose monophosphate (RuMP) pathway or dissimilated via the tetrahydromethanopterin (H4MPT) pathway. The deficiency of glycolysis and the TCA cycle indicate that strain TWE2 may be an obligate methylotroph. This is the first complete genome sequence of the genus Methylophilus. PMID:26253961

  17. Diversity of Denitrifying Bacteria in the San Francisco Bay

    Atluri, A.; Lee, J.; Francis, C. A.

    2012-12-01

    We compared the diversity of communities of denitrifying bacteria from the San Francisco Bay to investigate whether environmental factors affect diversity. To do this, we studied the sequence diversity of the marker gene nirK. nirK codes for the enzyme nitrite reductase which helps reduce nitrite to nitric oxide, an important step in denitrification. Sediment samples were collected spatially from five different locations and temporally during the four different seasons along a salinity gradient in the bay. After collecting samples and extracting DNA from them, we used PCR to amplify our gene of interest, created clone libraries for sequencing, and compared phylogenetic trees from the different communities. Based on several phylogenetic analyses on our tree and environments, we saw that denitrifying bacteria from the North and Central Bay form distinct spatial clusters; Central Bay communities are very similar to each other, while communities from the North Bay are more distinct from each other and from communities in the Central Bay. Bacteria from site 8.1M (Carquinez Strait) showed the most cm-scale spatial diversity, and there was the most species richness during the winter. All this suggests that diversity of communities of denitrifying bacteria may be affected by spatial and temporal environmental factors.

  18. Experimental investigation of activities and tolerance of denitrifying bacteria under alkaline and reducing condition

    In the geological disposal system of TRU wastes, nitrogen generation by denitrifying bacteria could provide significant impact on the assessment of this system, because nitrate contained in process concentrated liquid waste might be electron acceptor for denitrifying bacteria. In this study, the activities and tolerance of denitrifying under disposal condition were investigated. Pseudomonas denitrificans as denitrifying bacteria was used. The results showed that Pseudomonas denitrificans had activity under reducing condition, but under high pH condition (pH>9.5), the activity of Pseudomonas denitrificans was not detected. It is possible that the activity of Pseudomonas denitrificans would be low under disposal condition. (author)

  19. Rapid in vitro regeneration method for Moringa oleifera and performance evaluation of field grown nutritionally enriched tissue cultured plants

    Saini, R. K.; Shetty, N. P.; P.Giridhar; Ravishankar, G.A

    2012-01-01

    The present investigations were attempted to develop the rapid in vitro micropropagation protocol of Moringa oleifera (Variety-PKM-1) from nodal sections of young, aseptically grown seedlings. Benzyladenine (BA) at 4.44 μM was found to be optimal in producing on maximum an average of 9.0 ± 1.0 axillary shoots per explant after 15 days of inoculation. A high multiplication rate was established through routine sub culturing of nodal sections explanted from in vitro shoot cultures. In vitro root...

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

    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)

  1. D/H fractionation in lipids of facultative and obligate denitrifying and sulfate reducing bacteria

    Osburn, M. R.; Sessions, A. L.

    2012-12-01

    The hydrogen isotopic composition of lipids has been shown to vary broadly in both cultured bacteria and in environmental samples. Culturing studies have indicated that this variability may primarily reflect metabolism; however, the limited number of organisms studied thus far prevents application of these trends to interpretation of environmental samples. Here we report D/H fractionations in anaerobic bacteria, including both facultative and obligate anaerobic organisms with a range of electron donors, acceptors, and metabolic pathways. Experiments using the metabolically flexible alphaproteobacterium Paracoccus denitrificans probe particular central metabolic pathways using a range of terminal electron acceptors. While a large range of δD values has been observed during aerobic metabolism, denitrifying cultures produce a more limited range in δD values that are more similar to each other than the corresponding aerobic culture. Data from the sulfate reducing bacteria Desulfobacterium autotrophicum and Desulfobacter hydrogenophilus indicate that chemolithoautotrophy and anaerobic heterotrophy can produce similar δD values, and are similar between bacteria despite differing metabolic pathways. These results suggest that the fractionation of D/H depends both on the specific metabolic pathway and the electron acceptor. While this is not inconsistent with previous studies, it suggests the simple correspondence between δD and metabolism previously understood from aerobic bacteria is not universally applicable.

  2. Criteria and Methodology for Identifying Respiratory Denitrifiers

    Mahne, I.; Tiedje, J.M.

    1995-01-01

    Respiratory denitrification is not always adequately established when bacteria are characterized. We have tested a simple method that allows one to evaluate whether the two necessary criteria to claim denitrification have been met, namely, that N(inf2) or N(inf2)O is produced from nitrate or nitrite and that this reduction is coupled to a growth yield increase. Microorganisms were cultured in sealed tubes under a helium headspace and in the presence of 0, 2, 4, 7, and 10 mM nitrate or nitrite...

  3. Comparative analysis of 16S rRNA and amoA genes from archaea selected with organic and inorganic amendments in enrichment culture.

    Xu, Mouzhong; Schnorr, Jon; Keibler, Brandon; Simon, Holly M

    2012-04-01

    We took advantage of a plant-root enrichment culture system to characterize mesophilic soil archaea selected through the use of organic and inorganic amendments. Comparative analysis of 16S rRNA and amoA genes indicated that specific archaeal clades were selected under different conditions. Three amoA sequence clades were identified, while for a fourth group, identified by 16S rRNA gene analysis alone and referred to as the "root" clade, we detected no corresponding amoA gene. The amoA-containing archaea were present in media with either organic or inorganic amendments, whereas archaea representing the root clade were present only when organic amendment was used. Analysis of amoA gene abundance and expression, together with nitrification-coupled growth assays, indicated potential growth by autotrophic ammonia oxidation for members of two group 1.1b clades. Increased abundance of one of these clades, however, also occurred upon the addition of organic amendment. Finally, although amoA-containing group 1.1a archaea were present in enrichments, we detected neither expression of amoA genes nor evidence for nitrification-coupled growth of these organisms. These data support a model of a diverse metabolic community in mesophilic soil archaea that is just beginning to be characterized. PMID:22267662

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

    Gonzlez-Toril, E.; Amils, R.; Delmas, R. J.; Petit, J.-R.; Komrek, J.; Elster, J.

    2009-01-01

    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.

  5. Azoarcus taiwanensis sp. nov., a denitrifying species isolated from a hot spring.

    Lee, Duu-Jong; Wong, Biing-Teo; Adav, Sunil S

    2014-02-01

    The strain NSC3(T), a novel, facultative, chemolithotrophic, denitrifying, alkaliphilic, sulfide-oxidizing bacterium isolated from a hot spring in Yang-Ming Mountain, Taiwan, was Gram negative, rod shaped, and motile by single polar flagella and grew facultatively by adopting a denitrifying metabolism. The 16S rRNA sequence analysis revealed that strain NSC3(T) belongs to beta subclass of the Proteobacteria and most closely related to Azoarcus evansii KB740(T) (95.44 %), Azoarcus toluvorans Td-21(T) (95.21 %), Azoarcus tolulyticus Tol-4(T) (95.08 %), and Azoarcus toluclasticus MF63(T) (94.94 %). The phylogenetic analyses based on 16S rRNA gene sequences indicated that the strain NSC3(T) formed a distinct lineage in the Betaproteobacteria and that it exhibited the highest level of sequence similarity with species of the genera Azoarcus (95.28-93.13 %). The major fatty acids of the type strain were C16:0 (26.9 %), C16:1w7c (28.9 %), C18:0 (9.6 %), and C18:1w7c/w6c (29.9 %). The DNA G+C content of genomic DNA was 63.7 mol%. On the basis of the 16S rRNA sequence similarity, phenotypic and genotypic characteristics, and chemotaxonomic data, the strain NSC3(T) could be differentiated from other species of the genus Azoarcus. Therefore, strain NSC3(T) (equal to BCRC 80111(T) and DSM 24109(T)) is proposed as a novel species in genus Azoarcus, for which the name Azoarcus taiwanensis sp. nov. is proposed. The strain NSC3(T) is deposited in Bioresource Collection and Research Center, Taiwan, under the reference number BCRC 80111(T), and German Collection of Microorganisms and Cell Cultures, Germany (DSMZ), with DSM 24109(T). PMID:23695778

  6. Molecular and Stable Isotope Investigation of Nitrite Respiring Bacterial Communities Capable of Anaerobic Ammonium Oxidation (ANAMMOX) and Denitrifying Anaerobic Methane Oxidation (DAMO) in Nitrogen Contaminated Groundwater

    Song, B.; Hirsch, M.; Taylor, J.; Smith, R. L.; Repert, D.; Tobias, C. R.

    2010-12-01

    Anaerobic ammonium oxidation (ANAMMOX) and denitrifying anaerobic methane oxidation (DAMO) are two recently discovered N2 production pathways in the microbial nitrogen cycle. ANAMMOX has been relatively well investigated in various aquatic ecosystems, while DAMO has been examined only in freshwater wetlands. However, neither ANAMMOX nor DAMO have been studied in groundwater ecosystems as microbial N removal processes where they could compliment or compete with denitrification to remediate N contaminated aquifers. Thus, we conducted molecular and stable isotope analyses to detect and measure ANAMMOX and DAMO in a nitrogen contaminated aquifer on Cape Cod, Massachusetts. The study site has a plume of nitrogen contaminated groundwater as a result of continuous discharge of treated wastewater over 60 years. Groundwater was collected from multiport sampling devices installed at two sites, near the waste-water disposal location (A) and more than 3 km down gradient (B) along the contamination plume. Biomass was collected from water samples for DNA extraction and 15N tracer incubation experiments. PCR with specific 16S rRNA gene primers detected the presence of ANAMMOX and DAMO bacteria at both sites. Phylogenetic analysis of 16S rRNA genes revealed that the ANAMMOX community at site A was most associated with Kuenenia spp. while site B had a community more closely related to Brocadia spp. The DAMO communities at the two sites were quite different based on 16S rRNA gene analysis. The communities at site B are closely associated with Candidatus “Methylomirabilis oxyfera”, which is the first enriched DAMO culture. Most of the 16S rRNA sequences detected in site A were related to those found in other DAMO enrichment cultures established from a eutrophic ditch sediment. In order to determine active members of ANAMMOX communities, the transcriptional expression of hydrazine oxidase (hzo) and hydrazine hydrolase (hh) genes was examined at both sites. In addition, 15N tracer incubation experiments were used to measure the rates of ANAMMOX and denitrification. ANAMMOX was found to be higher than denitrification at site A where ANAMMOX accounted for 60% of the 15N2 production. In contrast, denitrification was higher than ANAMMOX at site B where Methylomirabilis spp. were found. Thus, this study clearly demonstrates the potential importance of ANAMMOX and DAMO in the nitrogen removal from groundwater and suggests that detailed characterization of the processes under in situ subsurface conditions could provide new information regarding the ecology of these microbes.

  7. Dynamics of Soil Denitrifier Populations: Relationships between Enzyme Activity, Most-Probable-Number Counts, and Actual N Gas Loss †

    Martin, Kendall; Parsons, Laura L.; Murray, Robert E.; Smith, M. Scott

    1988-01-01

    To better understand temporal variability in soil denitrification, denitrifying enzyme activity (DEA) and denitrifier populations (as determined by most-probable-number [MPN] counts) were measured in field and laboratory experiments. Measurements of DEA and MPN provided highly contradictory indications of denitrifier dynamics. In laboratory incubations, under conditions favoring active denitrification, the synthesis of new denitrifying enzymes and the actual amount of denitrification were clo...

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

    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 (186114ml/L-WW/d compared to 370C (65130ml/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.

  9. Pandoraea oxalativorans sp. nov., Pandoraea faecigallinarum sp. nov. and Pandoraea vervacti sp. nov., isolated from oxalate-enriched culture.

    Sahin, Nurettin; Tani, Akio; Kotan, Recep; Sedlcek, Ivo; Kimbara, Kazuhide; Tamer, Abdurrahman U

    2011-09-01

    Five isolates, designated TA2, TA4, TA25(T), KOx(T) and NS15(T) were isolated in previous studies by enrichment in mineral medium with potassium oxalate as the sole carbon source and were characterized using a polyphasic approach. The isolates were Gram-reaction-negative, aerobic, non-spore-forming rods. Phylogenetic analyses based on 16S rRNA and DNA gyrase B subunit (gyrB) gene sequences confirmed that the isolates belonged to the genus Pandoraea and were most closely related to Pandoraea sputorum and Pandoraea pnomenusa (97.2-99.7?% 16S rRNA gene sequence similarity). The isolates could be differentiated from their closest relatives on the basis of several phenotypic characteristics. The major cellular fatty acid profiles of the isolates comprised C??:?, C??:??7c, C??:? cyclo and summed feature 3 (C??:??7c and/or iso-C??:? 2-OH). On the basis of DNA-DNA hybridization studies and phylogenetic analyses, the isolates represent three novel species within the genus Pandoraea, for which the names Pandoraea oxalativorans sp. nov. (TA25(T) ?=?NBRC 106091(T) ?=?CCM 7677(T) ?=?DSM 23570(T)), Pandoraea faecigallinarum sp. nov. (KOx(T) ?=?NBRC 106092(T) ?=?CCM 2766(T) ?=?DSM 23572(T)) and Pandoraea vervacti sp. nov. (NS15(T) ?=?NBRC 106088(T) ?=?CCM 7667(T) ?=?DSM 23571(T)) are proposed. PMID:20952546

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

    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.

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

    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.

  12. Elimination of pharmaceuticals in single- and three-stage pre-denitrifying MBBR

    Polesel, Fabio; Torresi, Elena; Loreggian, L.; Escola Casas, Monica; Bester, Kai; Plsz, Benedek G.

    This study investigated the elimination of pharmaceuticals in pre-denitrifying moving bed biofilm reactors (MBBRs) in single- and three-stage configurations. Under batch conditions, biotransformation and retransformation of two pharmaceuticals (trimethoprim, sulfamethoxazole) occurred at comparable...... or higher rates than in denitrifying activated sludge. Based on estimated rate coefficients, concentrations in continuously operated MBBRs were predicted....

  13. Isotope enrichment

    In this chapter of the textbook for chemists the isotope enrichment methods are overviewed. The subsections are: General characterization of the methods used for isotope enrichment, Distillation and chemical exchange, Electrochemical methods, Diffusion methods, Isotope enrichment through centrifugation, Single-stage methods, Cascades

  14. Determination of the cause of the symptoms on yellow yam (Dioscorea cayenensis Lam.) leaf tissue and their eradication, enriching the culture medium and using techniques of meristem culture, thermo and chemotherapy on in vitro conditions

    Yams (Dioscorea spp) has been cultivated for exportation in Costa Rica, in North Huetar region. In vitro culture technique has been used for multiplying planting material for many advantages. However, cleaning of viruses that affect has been ineffective. Viruses such as: the potyvirus, potexvirus, cucumovirus . Methods like meristem culture, chemotherapy, thermotherapy and combinations of these have been used for the elimination of virus in plant species. The plants were evaluated in indexing assays, observing symptoms, serological methods and electron microscopy, among others. Other problems that have been affecting in vitro plant are deficient culture media in some nutrient. The presence of some abnormal characteristics in leaf tissue was determined whether have been caused by a virus or a nutritional deficiency in the culture medium. The presence of the virus has tried to find using ELISA and electron microscopy. Tests meristem culture, thermotherapy and chemotherapy have been made for the eradication of a possible virus; which have been assessed by observation of symptomatology and ELISA. The efficiency of the culture medium was evaluated to enrich it with nitrogen or excess iron. None of the suspected virus found in ELISA tests. Filaments are presumably viral particles were found through analysis of ultrastructure, as well as alterations in chloroplasts which indicated the presence of a pathogen or toxicity. Thermotherapy and chemotherapy with the concentration of 40 mg/L of ribavirin have been the most effective for the elimination of symptoms in virus eradication treatments. Assessments nutrient concentrations have shown that the differences between the various treatments used were undetectable. The symptoms presented were caused, according to the conclusions, by a virus which should preferably deal with thermotherapy. (author)

  15. Functional gene pyrosequencing reveals core proteobacterial denitrifiers in boreal lakes.

    Saarenheimo, Jatta; Tiirola, Marja Annika; Rissanen, Antti J

    2015-01-01

    Denitrification is an important microbial process in aquatic ecosystems that can reduce the effects of eutrophication. Here, quantification and pyrosequencing of nirS, nirK, and nosZ genes encoding for nitrite and nitrous oxide reductases was performed in sediment samples from four boreal lakes to determine the structure and seasonal stability of denitrifying microbial populations. Sediment quality and nitrate concentrations were linked to the quantity and diversity of denitrification genes, the abundance of denitrifying populations (nirS and nosZ genes) correlated with coupled nitrification-denitrification (Dn), and the denitrification of the overlying water NO3 (-) (Dw) correlated with the nirS/nirK ratio. The number of core nirS, nirK, and nosZ operational taxonomical units (OTUs) was low (6, 7, and 3, respectively), and most of these core OTUs were shared among the lakes. Dominant nirK sequences matched best with those of the order Rhizobiales, which was one of the main bacterial orders present in the sediment microbiomes, whereas the dominant nirS sequences were affiliated with the order Burkholderiales. Over half of the nosZ sequences belonged to a single OTU of the order Burkholderiales, but coupled nitrification-denitrification rate correlated with another dominant nosZ OTU assigned to the order Rhodospirillales. The study indicates that a few core proteobacterial clusters may drive denitrification in boreal lake sediments, as the same Alpha- and Betaproteobacteria denitrifier clusters were present in different lakes and seasons. PMID:26191058

  16. Effects of inorganic salts on denitrifying granular sludge: The acute toxicity and working mechanisms.

    Wang, Ru; Zheng, Ping; Ding, A-Qiang; Zhang, Meng; Ghulam, Abbas; Yang, Cheng; Zhao, He-Ping

    2016-03-01

    It is highly significant to investigate the toxicity of inorganic salts to denitrifying granular sludge (DGS) and its mechanism since the application of high-rate denitrification is seriously limited in the treatment of saline nitrogen-rich wastewaters. The batch experiments showed that the IC50 (half inhibition concentration) and LC50 (half lethal concentration) of NaCl, Na2SO4 and Na3PO4 on DGS were 11.46, 21.72, 7.46g/L and 77.35, 100.58, 67.92g/L respectively. Based on the analysis of specific denitrifying activity, the live cell percentage, the cell structure, and the DNA leakage, the toxicity of low salinity was ascribed to the inhibition of denitrifying activity and the toxicity of high salinity was ascribed to both the inhibition of denitrifying activity and the lethality of denitrifying cell. PMID:26773376

  17. Modeling denitrifying sulfide removal process using artificial neural networks

    The denitrifying sulfide removal (DSR) process has complex interactions between autotrophic and heterotrophic denitrifers; thus, constructing a detailed mechanistic model and proper control architecture is difficult. Artificial neural networks (ANNs) are capable of inferring the complex relationships between input and output process variables without a detailed characterization of the mechanisms governing the process. This work presents a novel ANN that accurately predicts the steady-state performance of an expended granular sludge bed (EGSB)-DSR bioreactor for nitrite denitrification and the complete DSR process. The proposed ANN shows that at a threshold hydraulic retention time (HRT) < 7 h, influent sulfide concentration markedly affects reactor performance.

  18. Chloramphenicol Inhibition of Denitrifying Enzyme Activity in Two Agricultural Soils

    Murray, Robert E.; Knowles, Roger

    1999-01-01

    Chloramphenicol, at concentrations greater than 0.1 g/liter (0.3 mM), inhibited the denitrifying enzyme activity (DEA) of slurries of humisol and sandy loam soils by disrupting the activity of existing nitrate reductase enzymes. When the concentration of chloramphenicol was increased from 0.1 to 2.0 g/liter (6.0 mM), the rate of nitrite production from nitrate decreased by 25 to 46%. The rate of NO production from nitrate decreased by 20 to 39%, and the rate of N2O production from nitrate, in...

  19. Warming-induced changes in denitrifier community structure modulate the ability of phototrophic river biofilms to denitrify

    Boulêtreau, Stéphanie, E-mail: stephanie.bouletreau@univ-tlse3.fr [Université de Toulouse, UPS, INP, EcoLab (Laboratoire Ecologie Fonctionnelle et Environnement), 118 route de Narbonne, F-31062 Toulouse (France); CNRS, EcoLab, F-31062 Toulouse (France); Lyautey, Emilie [Université de Toulouse, UPS, INP, EcoLab (Laboratoire Ecologie Fonctionnelle et Environnement), 118 route de Narbonne, F-31062 Toulouse (France); CNRS, EcoLab, F-31062 Toulouse (France); Dubois, Sophie [Université de Bordeaux, EPOC - OASU, UMR 5805, Station Marine d' Arcachon, 2 rue du Professeur Jolyet, 33120 Arcachon (France); Compin, Arthur [Université de Toulouse, UPS, INP, EcoLab (Laboratoire Ecologie Fonctionnelle et Environnement), 118 route de Narbonne, F-31062 Toulouse (France); CNRS, EcoLab, F-31062 Toulouse (France); Delattre, Cécile; Touron-Bodilis, Aurélie [EDF Recherche et Développement, LNHE (Laboratoire National d' Hydraulique et Environnement), 6 quai Watier, F-78401 Chatou (France); Mastrorillo, Sylvain [Université de Toulouse, UPS, INP, EcoLab (Laboratoire Ecologie Fonctionnelle et Environnement), 118 route de Narbonne, F-31062 Toulouse (France); CNRS, EcoLab, F-31062 Toulouse (France); Garabetian, Frédéric [Université de Bordeaux, EPOC - OASU, UMR 5805, Station Marine d' Arcachon, 2 rue du Professeur Jolyet, 33120 Arcachon (France)

    2014-01-01

    Microbial denitrification is the main nitrogen removing process in freshwater ecosystems. The aim of this study was to show whether and how water warming (+ 2.5 °C) drives bacterial diversity and structuring and how bacterial diversity affects denitrification enzymatic activity in phototrophic river biofilms (PRB). We used water warming associated to the immediate thermal release of a nuclear power plant cooling circuit to produce natural PRB assemblages on glass slides while testing 2 temperatures (mean temperature of 17 °C versus 19.5 °C). PRB were sampled at 2 sampling times during PRB accretion (6 and 21 days) in both temperatures. Bacterial community composition was assessed using ARISA. Denitrifier community abundance and denitrification gene mRNA levels were estimated by q-PCR and qRT-PCR, respectively, of 5 genes encoding catalytic subunits of the denitrification key enzymes. Denitrification enzyme activity (DEA) was measured by the acetylene-block assay at 20 °C. A mean water warming of 2.5 °C was sufficient to produce contrasted total bacterial and denitrifier communities and, therefore, to affect DEA. Indirect temperature effect on DEA may have varied between sampling time, increasing by up to 10 the denitrification rate of 6-day-old PRB and decreasing by up to 5 the denitrification rate of 21-day-old PRB. The present results suggest that indirect effects of warming through changes in bacterial community composition, coupled to the strong direct effect of temperature on DEA already demonstrated in PRB, could modulate dissolved nitrogen removal by denitrification in rivers and streams. - Highlights: •We produced river biofilms in 2 mean temperature conditions: 17 vs 19.5 °C. •We compared their denitrifiers' structuring and functioning in 6d- and 21d-old biofilms. •A difference of 2.5 °C produced contrasted denitrifier communities. •The indirect temperature effect on denitrification activity shifted between biofilm age. •Warming impact strongly depends on the bacterial successional trajectory.

  20. Warming-induced changes in denitrifier community structure modulate the ability of phototrophic river biofilms to denitrify

    Microbial denitrification is the main nitrogen removing process in freshwater ecosystems. The aim of this study was to show whether and how water warming (+ 2.5 °C) drives bacterial diversity and structuring and how bacterial diversity affects denitrification enzymatic activity in phototrophic river biofilms (PRB). We used water warming associated to the immediate thermal release of a nuclear power plant cooling circuit to produce natural PRB assemblages on glass slides while testing 2 temperatures (mean temperature of 17 °C versus 19.5 °C). PRB were sampled at 2 sampling times during PRB accretion (6 and 21 days) in both temperatures. Bacterial community composition was assessed using ARISA. Denitrifier community abundance and denitrification gene mRNA levels were estimated by q-PCR and qRT-PCR, respectively, of 5 genes encoding catalytic subunits of the denitrification key enzymes. Denitrification enzyme activity (DEA) was measured by the acetylene-block assay at 20 °C. A mean water warming of 2.5 °C was sufficient to produce contrasted total bacterial and denitrifier communities and, therefore, to affect DEA. Indirect temperature effect on DEA may have varied between sampling time, increasing by up to 10 the denitrification rate of 6-day-old PRB and decreasing by up to 5 the denitrification rate of 21-day-old PRB. The present results suggest that indirect effects of warming through changes in bacterial community composition, coupled to the strong direct effect of temperature on DEA already demonstrated in PRB, could modulate dissolved nitrogen removal by denitrification in rivers and streams. - Highlights: •We produced river biofilms in 2 mean temperature conditions: 17 vs 19.5 °C. •We compared their denitrifiers' structuring and functioning in 6d- and 21d-old biofilms. •A difference of 2.5 °C produced contrasted denitrifier communities. •The indirect temperature effect on denitrification activity shifted between biofilm age. •Warming impact strongly depends on the bacterial successional trajectory

  1. Dynamic Metabolic Modeling of Denitrifying Bacterial Growth: The Cybernetic Approach

    Song, Hyun-Seob; Liu, Chongxuan

    2015-06-29

    Denitrification is a multistage reduction process converting nitrate ultimately to nitrogen gas, carried out mostly by facultative bacteria. Modeling of the denitrification process is challenging due to the complex metabolic regulation that modulates sequential formation and consumption of a series of nitrogen oxide intermediates, which serve as the final electron acceptors for denitrifying bacteria. In this work, we examined the effectiveness and accuracy of the cybernetic modeling framework in simulating the growth dynamics of denitrifying bacteria in comparison with kinetic models. In four different case studies using the literature data, we successfully simulated diauxic and triauxic growth patterns observed in anoxic and aerobic conditions, only by tuning two or three parameters. In order to understand the regulatory structure of the cybernetic model, we systematically analyzed the effect of cybernetic control variables on simulation accuracy. The results showed that the consideration of both enzyme synthesis and activity control through u- and v-variables is necessary and relevant and that uvariables are of greater importance in comparison to v-variables. In contrast, simple kinetic models were unable to accurately capture dynamic metabolic shifts across alternative electron acceptors, unless an inhibition term was additionally incorporated. Therefore, the denitrification process represents a reasonable example highlighting the criticality of considering dynamic regulation for successful metabolic modeling.

  2. A method for measuring the anoxic biodegradability under denitrifying conditions.

    Vzquez-Rodrguez, Gabriela A; Beltrn-Hernndez, Rosa Icela; Lucho-Constantino, Carlos Alexander; Blasco, Jos Luis

    2008-04-01

    A test for assessing the anoxic biodegradability of organic compounds under denitrifying conditions is proposed. The method is based on the recovery and quantification of the CO2 produced, which is evidence of complete biodegradation of the test compound (added as the sole carbon source). The tests were carried out in a mineral medium, with nitrate as electron acceptor. Whole lake sediments, sediment extracts and a commercial inoculum were assayed as a possible inoculum source by means of glucose biodegradability tests. It was found that the sediment extracts constitute a suitable and environmentally-relevant inoculum source, since they add non-significant amounts of carbon to the tests. Two xenobiotic compounds, namely, aniline and phenol, were tested in the aforementioned conditions as well as in a standard aerobic biodegradability test. Both aniline and phenol attained a biodegradation level higher than 60% in a short time period (biodegradable in denitrifying environments. Nevertheless, the kinetics obtained in the anoxic test were slower than in aerobic conditions, and even suggested the accumulation of intermediate metabolites in the case of phenol. The results of this study indicate that the fate of xenobiotic compounds under anoxic conditions differs from that observed in an oxic environment, and therefore it should be considered by standard biodegradability testing procedures. PMID:18096201

  3. Enrichment of denitrifying methanotrophic bacteria for application after direct low-temperature anaerobic sewage treatment

    Kampman, C.; Hendrickx, T.L.G.; Luesken, F.; van Alen, T. A.; Op den Camp, H. J. M.; Jetten, M. S. M.; Zeeman, G.; Buisman, C.J.N.; Temmink, B.G.

    2012-01-01

    Despite many advantages of anaerobic sewage treatment over conventional activated sludge treatment, it has not yet been applied in temperate zones. This is especially because effluent from low-temperature anaerobic treatment contains nitrogen and dissolved methane. The presence of nitrogen and methane offers the opportunity to develop a reactor in which methane is used as electron donor for denitrification. Such a reactor could be used in a new concept for low-temperature anaerobic sewage tre...

  4. Enrichment of denitrifying phosphorus accumulating organisms (DPAOs) in a continuous flow laboratory scale plant

    Enhanced Biological Phosphate Removal (EBPR) is a well established method for efficient removal of phosphate during wastewater treatment by using biological instead of chemical phenomena. EBPR is currently implemented at a number of Waste Water Treatment Plants (WWTPs) and usually phosphate removal occurs by recirculating activated sludge between an anaerobic and a aerobic tank. (Author)

  5. The social cost of coastal erosion. Using cultural theory to enrich the interpretation of stated preference data.

    Kontogianni, A.; Tourkolias, C.; Vousdoukas, M.; Skourtos, M.

    2012-04-01

    Natural coastal processes are to a great extent modified by proximity to man-made structures. Engineered interventions, port facilities, housing and industrial infrastructure, all can increase the coastline fluctuations significantly relative to those along a long unobstructed coastline. As a consequence, coastlines are increasingly exposed to coastal erosion, a phenomenon defined as the encroachment of land by the sea after averaging over a period, which is sufficiently long to eliminate the impacts of weather, storm events and local sediment dynamics. In order to provide cost effective management of coastal erosion it is crucial to estimate both the benefits and costs associated with various management alternatives. The initiatives on Integrated Coastal Zone Manegment in Europe, but also the upcoming Marine Strategy Framwork Directive would benefit greatly from a proliferation of socioeconomic information to assist decision makers who must weigh the impacts of various types of coastal improvement and the cost of beach protection/restoration. In that spirit, the objective of the present research is to report the results of a survey undertaken in two resort beaches on the island of Lesvos (Greece), designed to estimate public preferences for avoiding coastal erosion. A mixed methodological approach is employed by combining an open-ended contingent valuation survey with cultural theory of risk perception. The empirical models to analyze individual choices of erosion control programs and the associated welfare measures are presented, followed by the discussion of model specification and estimation issues, and the results of the data analysis. Some concluding remarks are then presented. By choosing this approach we aim at improving our understanding of preference structure for avoiding public risk, accepted level of risk and perceptions thereof. The framework can also be used for assessing the social cost of extreme weather events such as storm surges in the coastal zone, to get an insight for insurance values.

  6. A Fungal Cytochrome P-450nor Confers Denitrifying Ability to Tobacco By-2 Cells

    Babiker M.A. Abdel-Banat

    2008-01-01

    Full Text Available Reactive nitrogen gases progressively contribute to the global warming. Development of gas-gas denitrifying plants that can efficiently reduce reactive nitrogen gases to dinitrogen (N2 could help to mitigate the effect of these gases. Taking the advances in gene manipulation technology, tobacco BY-2 cells were transformed with the fungus Cylindrocarpon tonkinense cytochrome P-450nor2 (Cnor2 gene. The product of this gene acts as nitric oxide reductase (nor. Transgenic BY-2 cell clones cultured in 15N-labelled nitrate (15NO3- actively evolved 15N2O gas up to 35-folds compared to the wild-type cells. In 15N-labelled ammonium (15NH4+, the transgenic and wild-type cells produced comparable amounts of 15N2O. This indicates that ammonium is not a direct substrate for nor and the small amount of N2O observed may be due to the nitrification of ammonium to nitrite. Addition of tungstate (a nitrate reductase inhibitor and cyanide to the transgenic cell cultures strongly inhibited 15N2O production. Activity of nor enzyme was also confirmed by in vitro activity assay. These observations together suggest that Cnor2 is actively expressed and enhanced the reduction of nitrate to N2O in plant cells. This finding indicates that plant cells are capable to tackle the denitrification pathway.

  7. Elemental sulfur formation and nitrogen removal from wastewaters by autotrophic denitrifiers and anammox bacteria.

    Liu, Chunshuang; Zhao, Dongfeng; Yan, Laihong; Wang, Aijie; Gu, Yingying; Lee, Duu-Jong

    2015-09-01

    Elemental sulfur (S(0)) formation from and nitrogen removal on sulfide, nitrate and ammonium-laden wastewaters were achieved by denitrifying ammonium oxidation (DEAMOX) reactor with autotrophic denitrifiers and anaerobic ammonium oxidation (anammox) bacteria. The sulfide to nitrate ratio is a key process parameter for excess accumulation of S(0) and a ratio of 1.31:1 is a proposed optimum. The Alishewanella, Thauera and Candidatus Anammoximicrobium present respectively the autotrophic denitrifiers and anammox bacteria for the reactor. DEAMOX is demonstrated promising biological process for treating organics-deficient (S+N) wastewaters with excess S(0) production. PMID:26022701

  8. Enhanced performance of denitrifying sulfide removal process under micro-aerobic condition

    The denitrifying sulfide removal (DSR) process with bio-granules comprising both heterotrophic and autotrophic denitrifiers can simultaneously convert nitrate, sulfide and acetate into di-nitrogen gas, elementary sulfur and carbon dioxide, respectively, at high loading rates. This study determines the reaction rate of sulfide oxidized into sulfur, as well as the reduction of nitrate to nitrite, would be enhanced under a micro-aerobic condition. The presence of limited oxygen mitigated the inhibition effects of sulfide on denitrifier activities, and enhanced the performance of DSR granules. The advantages and disadvantages of applying the micro-aerobic condition to the DSR process are discussed.

  9. Greenhouse Gas Emission from In-situ Denitrifying Bioreactors

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

    2013-12-01

    Despite decades of concerted effort to mitigate nonpoint source nitrate (NO3-) pollution from agricultural lands, these efforts have not been sufficient to arrest eutrophication, which continues to be a serious and chronic problem. Two primary processes for removing excess NO3- from water are biological assimilation and denitrification. Denitrifying bacteria use NO3- as the electron acceptor for respiration in the absence of oxygen. Denitrification results in reduced forms of nitrogen, often dinitrogen gas (N2) but also nitrous oxide (N2O), an aggressive greenhouse gas (GHG). A promising solution to NO3- pollution is to intercept agricultural discharges with denitrifying bioreactors (DNBRs), though research has been limited to NO3- level reduction and omitted process mechanisms. DNBRs work by providing an anaerobic environment with plenty of organic matter (commonly woodchips) for denitrifying bacteria to flourish. While, initial results from bioreactor studies show that they can cost-effectively remove NO3-, GHG emission could be an unintended consequence. The study's goal is to determine how bioreactor design promotes microbial denitrification while limiting N2O production. It specifically focuses on expanding the body of knowledge concerning DNBRs in the areas of design implications and internal processes by measuring intermediate compounds and not solely NO3-. Nutrient samples are collected at inflow and outflow structures and tested for NO3- and nitrite (NO2-). Dissolved and headspace gas samples are collected and tested for N2O. Additional gas samples will be analyzed for naturally-occurring isotopic N2 to support proposed pathways. Designs will be analyzed both through the N2O/N2 production ratio and NO2- production caused by various residence times and inflow NO3- concentrations. High GHG ratios and NO2- production suggest non-ideal conditions or flow patterns for complete denitrification. NO3- reduction is used for comparison with previous studies. Few studies have focused on the reactions and products of DNBRs and instead note NO3- reductions in outflows as evidence that this should be used to treat agricultural discharges. It is, however, critical to understand all of the potential impacts that this treatment solution may have, which means identifying processes and design specifications that may affect them. This study addresses this lack of mechanistic understanding of DNBRs by means of sampling for incomplete products. Results will quantify gaseous products and internal processes and present a more complete picture of the potential for widespread use of DNBRs.

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

    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.

  11. Development and evaluation of a loop-mediated isothermal amplification assay combined with enrichment culture for rapid detection of very low numbers of Vibrio parahaemolyticus in seafood samples.

    Di, Huiling; Ye, Lei; Neogi, Sucharit Basu; Meng, Hecheng; Yan, He; Yamasaki, Shinji; Shi, Lei

    2015-01-01

    The aim of this study was to develop and evaluate a rapid and effective method to detect Vibrio parahaemolyticus, a leading pathogen causing seafood-borne gastroenteritis. A newly designed loop-mediated isothermal amplification (LAMP) assay including a short enrichment period was optimized. This assay correctly detected all the target strains (n=61) but none of the non-target strains (n=34). Very low numbers of V. parahaemolyticus (2 colony forming unit (CFU) per gram of seafood) could be detected within 3 h and the minimum time of the whole assay was only 5 h. Comparative screening of various seafood samples (n=70) indicated that the LAMP assay is superior to polymerase chain reaction (PCR) and conventional culture methods because it is more rapid and less complex. This highly sensitive LAMP assay can be applicable as the method of choice in large-scale and rapid screening of seafood and environmental samples to detect V. parahaemolyticus strains. PMID:25744462

  12. Uranium enrichment

    GAO was asked to address several questions concerning a number of proposed uranium enrichment bills introduced during the 100th Congress. The bill would have restructured the Department of Energy's uranium enrichment program as a government corporation to allow it to compete more effectively in the domestic and international markets. Some of GAO's findings discussed are: uranium market experts believe and existing market models show that the proposed DOE purchase of a $750 million of uranium from domestic producers may not significantly increase production because of large producer-held inventories; excess uranium enrichment production capacity exists throughout the world; therefore, foreign producers are expected to compete heavily in the United States throughout the 1990s as utilities' contracts with DOE expire; and according to a 1988 agreement between DOE's Offices of Nuclear Energy and Defense Programs, enrichment decommissioning costs, estimated to total $3.6 billion for planning purposes, will be shared by the commercial enrichment program and the government

  13. Microbial diversity in methanogenic hydrocarbon-degrading enrichment cultures isolated from a water-flooded oil reservoir (Dagang oil field, China)

    Jimnez, Nria; Cai, Minmin; Straaten, Nontje; Yao, Jun; Richnow, Hans H.; Krger, Martin

    2015-04-01

    Microbial transformation of oil to methane is one of the main degradation processes taking place in oil reservoirs, and it has important consequences as it negatively affects the quality and economic value of the oil. Nevertheless, methane could constitute a recovery method of carbon from exhausted reservoirs. Previous studies combining geochemical and isotopic analysis with molecular methods showed evidence for in situ methanogenic oil degradation in the Dagang oil field, China (Jimnez et al., 2012). However, the main key microbial players and the underlying mechanisms are still relatively unknown. In order to better characterize these processes and identify the main microorganisms involved, laboratory biodegradation experiments under methanogenic conditions were performed. Microcosms were inoculated with production and injection waters from the reservoir, and oil or 13C-labelled single hydrocarbons (e.g. n-hexadecane or 2-methylnaphthalene) were added as sole substrates. Indigenous microbiota were able to extensively degrade oil within months, depleting most of the n-alkanes in 200 days, and producing methane at a rate of 76 6 mol day-1 g-1 oil added. They could also produce heavy methane from 13C-labeled 2-methylnaphthalene, suggesting that further methanogenesis may occur from the aromatic and polyaromatic fractions of Dagang reservoir fluids. Microbial communities from oil and 2-methyl-naphthalene enrichment cultures were slightly different. Although, in both cases Deltaproteobacteria, mainly belonging to Syntrophobacterales (e.g. Syntrophobacter, Smithella or Syntrophus) and Clostridia, mostly Clostridiales, were among the most represented taxa, Gammaproteobacteria could be only identified in oil-degrading cultures. The proportion of Chloroflexi, exclusively belonging to Anaerolineales (e.g. Leptolinea, Bellilinea) was considerably higher in 2-methyl-naphthalene degrading cultures. Archaeal communities consisted almost exclusively of representatives of Methanomicrobia (mainly belonging to genera Methanosaeta and Methanoculleus). As both syntrophic Bacteria and methanogenic Archaea are abundant in Dagang, the studied areas of this oil field may have a significant potential to test the in situ conversion of oil into methane as a possible way to increase total hydrocarbon recovery.

  14. Performance of Denitrifying Microbial Fuel Cell with Biocathode over Nitrite.

    Zhao, Huimin; Zhao, Jianqiang; Li, Fenghai; Li, Xiaoling

    2016-01-01

    Microbial fuel cell (MFC) with nitrite as an electron acceptor in cathode provided a new technology for nitrogen removal and electricity production simultaneously. The influences of influent nitrite concentration and external resistance on the performance of denitrifying MFC were investigated. The optimal effectiveness were obtained with the maximum total nitrogen (TN) removal rate of 54.80 ± 0.01 g m(-3) d(-1). It would be rather desirable for the TN removal than electricity generation at lower external resistance. Denaturing gradient gel electrophoresis suggested that Proteobacteria was the predominant phylum, accounting for 35.72%. Thiobacillus and Afipia might benefit to nitrite removal. The presence of nitrifying Devosia indicated that nitrite was oxidized to nitrate via a biochemical mechanism in the cathode. Ignavibacterium and Anaerolineaceae was found in the cathode as a heterotrophic bacterium with sodium acetate as substrate, which illustrated that sodium acetate in anode was likely permeated through proton exchange membrane to the cathode. PMID:27047462

  15. Denitrifying sulfide removal process on high-tetracycline wastewater.

    Liu, Chunshuang; Xu, Jian; Lee, Duu-Jong; Yu, Daoyong; Liu, Lihong

    2016-04-01

    Antibiotics wastewater from tetracycline (TC) production unit can have high levels of chemical oxygen demand, ammonium and sulfate and up to a few hundreds of milligrams per liter of TC. Denitrifying sulfide removal (DSR) process is set up for simultaneously removal of sulfur, carbon and nitrogen from waters. The DSR process was for the first time studied for treating TC wastewaters. The TC stress has no adverse effects on removal rates of nitrate and acetate; however, it moderately deteriorated sulfide removal rates and S(0) accumulation rates when the concentration is higher than 100mgL(-1) TC. The Thauera sp., and Pseudomonas sp. present the heterotrophs and Sulfurovum sp. presented the autotroph for the present DSR reactions. The high tolerance of TC stress by the tested consortium was explained by the excess production of extracellular polymeric substances at high TC concentration, which can bind with TC for minimizing its inhibition effects. PMID:26810146

  16. Uranium enrichment

    Canada is the world's largest producer and exporter of uranium, most of which is enriched elsewhere for use as fuel in LWRs. The feasibility of a Canadian uranium-enrichment enterprise is therefore a perennial question. Recent developments in uranium-enrichment technology, and their likely impacts on separative work supply and demand, suggest an opportunity window for Canadian entry into this international market. The Canadian opportunity results from three particular impacts of the new technologies: 1) the bulk of the world's uranium-enrichment capacity is in gaseous diffusion plants which, because of their large requirements for electricity (more than 2000 kW·h per SWU), are vulnerable to competition from the new processes; 2) the decline in enrichment costs increases the economic incentive for the use of slightly-enriched uranium (SEU) fuel in CANDU reactors, thus creating a potential Canadian market; and 3) the new processes allow economic operation on a much smaller scale, which drastically reduces the investment required for market entry and is comparable with the potential Canadian SEU requirement. The opportunity is not open-ended. By the end of the century the enrichment supply industry will have adapted to the new processes and long-term customer/supplier relationships will have been established. In order to seize the opportunity, Canada must become a credible supplier during this century

  17. BIODEGRADATION OF AROMATIC COMPOUNDS UNDER MIXED OXYGEN/DENITRIFYING CONDITIONS: A REVIEW

    Bioremediation of aromatic hydrocarbons in groundwater and sediments is often limited by dissolved oxygen. Many aromatic hydrocarbons degrade very slowly or not at all under anaerobic conditions. Nitrate is a good alternative electron acceptor to oxygen, and denitrifying bacteria...

  18. Identification of active denitrifiers in full-scale nutrient removal wastewater treatment systems

    Mcllroy, Simon; Starnawska, Anna; Starnawski, Piotr; Saunders, Aaron M.; Nierychlo, Marta; Nielsen, Per Halkjær; Nielsen, Jeppe Lund

    2015-01-01

    Denitrification is essential to the removal of nitrogen from wastewater during treatment, yet an understanding of the diversity of the active denitrifying bacteria responsible in full-scale wastewater treatment plants (WWTPs) is lacking. In this study, stable-isotope probing (SIP) was applied in...... combination with microautoradiography (MAR)-fluorescence in situ hybridization (FISH) to identify previously unrecognized active denitrifying phylotypes in a full-scale WWTP with biological N and P removal. Acknowledging that different denitrifiers will have specific carbon source preferences, a fully 13C......-labelled complex substrate was used for SIP incubations, under nitrite-reducing conditions, in order to maximize the capture of the potentially metabolically diverse denitrifiers likely present. Members of the Rhodoferax, Dechloromonas, Sulfuritalea, Haliangium and Thermomonas were represented in the 16S rRNA gene...

  19. INHIBITION OF ALKYLBENZENE BIODEGRADATION UNDER DENITRIFYING CONDITIONS BY USING THE ACETYLENE BLOCK TECHNIQUE

    Addition of acetylene to microcosms simultaneously amended with nitrate and alkylbenzenes resulted in inhibition of the rate of alkylbenzene biodegradation under denitrifying conditions. Toluene, xylenes, and 1,2,4-trimethylbenzene were recalcitrant, whereas ethylbenzene was degr...

  20. Properties and electron transfer specificity of copper proteins from the denitrifier "Achromobacter cycloclastes".

    Liu, M. Y.; M. C. Liu; Payne, W. J.; LeGall, J.

    1986-01-01

    A blue copper protein (Mr 12,000) was purified from cells of "Achromobacter cycloclastes" grown as a denitrifier. When reduced, the blue copper protein transferred electrons to the copper protein nitrite reductase purified from the same cells, whereas a variety of cytochromes from denitrifiers failed to do so. Inclusion of a protease inhibitor, phenylmethylsulfonyl fluoride, in the buffers employed during preparation yielded purified blue copper protein with 18 more amino acid residues and tw...

  1. Immobilization of denitrifying bacteria with polymer carrier produced by radiation polymerization

    Polymer carriers were prepared from acrylamide, glycidyl methacrylate (GMA) and mixed monomer of glycidyl methacrylate (GMA) and 2-hydroxyethyl methacrylate (HEMA) by radiation polymerization. Two denitrifying strains of bacteria were immobilized into the polymer matrix. The denitrifying activity of immobilized cells with these carriers was related to the monomer composition of polymers. The optimum monomer composition was 50% in polyacrylamide and 5:15, 5:20, 10:20 % in poly (GMA-HEMA). (author)

  2. A Comparative Study of the Bacterial Community in Denitrifying and Traditional Enhanced Biological Phosphorus Removal Processes

    Lv, Xiao-Mei; Shao, Ming-Fei; Li, Chao-Lin; Li, Ji; Gao, Xin-lei; Sun, Fei-Yun

    2014-01-01

    Denitrifying phosphorus removal is an attractive wastewater treatment process due to its reduced carbon source demand and sludge minimization potential. Two lab-scale sequencing batch reactors (SBRs) were operated in alternating anaerobic-anoxic (A-A) or anaerobic-oxic (A-O) conditions to achieve denitrifying enhanced biological phosphate removal (EBPR) and traditional EBPR. No significant differences were observed in phosphorus removal efficiencies between A-A SBR and A-O SBR, with phosphoru...

  3. Spatial distribution of total, ammonia-oxidizing, and denitrifying bacteria in biological wastewater treatment reactors for bioregenerative life support

    Sakano, Yuko; Pickering, Karen D.; Strom, Peter F.; Kerkhof, Lee J.; Janes, H. W. (Principal Investigator)

    2002-01-01

    Bioregenerative life support systems may be necessary for long-term space missions due to the high cost of lifting supplies and equipment into orbit. In this study, we investigated two biological wastewater treatment reactors designed to recover potable water for a spacefaring crew being tested at Johnson Space Center. The experiment (Lunar-Mars Life Support Test Project-Phase III) consisted of four crew members confined in a test chamber for 91 days. In order to recycle all water during the experiment, an immobilized cell bioreactor (ICB) was employed for organic carbon removal and a trickling filter bioreactor (TFB) was utilized for ammonia removal, followed by physical-chemical treatment. In this study, the spatial distribution of various microorganisms within each bioreactor was analyzed by using biofilm samples taken from four locations in the ICB and three locations in the TFB. Three target genes were used for characterization of bacteria: the 16S rRNA gene for the total bacterial community, the ammonia monooxygenase (amoA) gene for ammonia-oxidizing bacteria, and the nitrous oxide reductase (nosZ) gene for denitrifying bacteria. A combination of terminal restriction fragment length polymorphism (T-RFLP), sequence, and phylogenetic analyses indicated that the microbial community composition in the ICB and the TFB consisted mainly of Proteobacteria, low-G+C gram-positive bacteria, and a Cytophaga-Flexibacter-Bacteroides group. Fifty-seven novel 16S rRNA genes, 8 novel amoA genes, and 12 new nosZ genes were identified in this study. Temporal shifts in the species composition of total bacteria in both the ICB and the TFB and ammonia-oxidizing and denitrifying bacteria in the TFB were also detected when the biofilms were compared with the inocula after 91 days. This result suggests that specific microbial populations were either brought in by the crew or enriched in the reactors during the course of operation.

  4. Uranium enrichment

    This report is a collection of chapters each outlining a different aspect of uranium enrichment. These include different technological procedures for enrichment with specific attention on: the Netherlands' ultracentrifuge; the market and economic situation; political and social implications as well as environmental problems; the history and decision-making at the governmental level which led to the treaty of Almelo; regional aspects; and finally a state-of-the-art evaluation. The report was composed solely on publicly-available data

  5. NirS-containing denitrifier communities in the water column and sediment of the Baltic Sea

    K. Jürgens

    2006-06-01

    Full Text Available The aim of this study was to compare structural differences in the denitrifying microbial communities along the environmental gradients observed in the water column and coastal sediments of the Baltic Sea. To link community structure and environmental gradients, denitrifier communities were analyzed by terminal restriction fragment length polymorphism (T-RFLP based on nirS as a functional marker gene for denitrification. NirS-type denitrifier community composition was further evaluated by phylogenetic analysis of nirS sequences from clone libraries. T-RFLP analysis indicated some overlap but also major differences of communities from the water column and the sediment. Shifts in community composition along the biogeochemical gradients were observed only in the water column while denitrifier communities were rather uniform within the upper 30 mm of the sediment. Specific terminal restriction fragments (T-RFs indicative for the sulfidic zone suggest the presence of nitrate-reducing and sulfide-oxidizing microorganisms that were previously shown to be important at the suboxic-sulfidic interface in the water column of the Baltic Sea. Phylogenetic analysis of nirS genes from the Baltic Sea and of sequences from marine habitats all over the world indicated distinct denitrifier communities that grouped mostly according to their habitat. We suggest that these subgroups of denitrifiers had developed after selection through several factors, i.e. their habitats (water column or sediment, impact by prevalent environmental conditions and isolation by large geographic distances between habitats.

  6. Improvement of sensitivity for Mycobacterium avium subsp. paratuberculosis (MAP) detection in bovine fecal samples by specific duplex F57/IC real-time and conventional IS900 PCRs after solid culture enrichment.

    Fawzy, Ahmad; Eisenberg, Tobias; El-Sayed, Amr; Zschck, Michael

    2015-04-01

    Mycobacterium avium subsp. paratuberculosis (MAP) is the etiological agent of Johne's disease in ruminants and a probable pathogen of Crohn's disease in humans. Accurate, cost-effective, and time-relevant diagnostics are the basis for efficient control programs. This study was conducted as an attempt to re-evaluate MAP detection improvement by coupling solid media enrichment to a more specific IS900 conventional PCR and a very specific F57/IC real-time PCR. In a spiking experiment, we investigated the improvement of molecular-based MAP detection in feces after a culture-based enrichment step into Herrold's egg yolk media with mycobactin J (HEYM-MJ) for different time intervals, when compared to traditional culture. Detection limit of culture was 0.33??10(4)bacteria??g(-1) (33CFUg(-1)), while that of IS900 PCR when coupled with an enrichment step for 2, 4, and 6weeks was 0.33??10(5) (0.33??10(3)CFUg(-1)), 0.33??10(4) (33CFUg(-1)), and 33 (>3.3CFUg(-1)) bacteria??g(-1), respectively. Whereas the detection limits of F57/IC real-time PCR after the enrichment step for the same time intervals were 0.33??10(5) (0.33??10(3)CFUg(-1)), 0.33??10(3) (3.3CFUg(-1)), and 33 (>3.3CFUg(-1)) bacteria??g(-1), respectively. Altogether, enrichment of bovine fecal samples into solid media increased the sensitivity of specific molecular detection of MAP using IS900 conventional PCR and duplex F57/IC real-time PCR and offers an expedited and accurate alternative for MAP detection in bovine feces. Validation of these results is further recommended using field bovine fecal samples. PMID:25716216

  7. [NO3-/NO2- inhibits sulfate-reducing activity of the enrichment culture of sulfate-reducing prokaryotes from an off-shore oil reservoir at Bohai Bay, China].

    Liu, Hong-Yu; Shi, Rong-Jiu; Zhang, Ying; Shi, Zhen-Guo; Zhang, Ying-Yue; Yu, Liang; Zhang, Xiao-Bo; Tan, Tao

    2014-08-01

    Long-term injection of sulfate-rich water into oil reservoirs stimulates the proliferation of sulfate-reducing prokaryotes (SRP) therein and results in production of a great amount of H2S, leading to souring in oil reservoirs and related environmental problems. In this study, we first, using modified API RP 38 medium, enriched SRP present in production water from a producing well at Bohai Bay, China, and then examined the inhibitory effects of nitrate or nitrite on sulfate reduction activity of the SRP. Results showed that the enriched SRP culture exhibited a high sulfate reduction activity as indicated by a sulfate-reducing rate of 10.4 mmol SO4(2-) x d(-1) x g(-1) dry cell. In presence of 0.4, 0.8, 1.8, and 4.2 mmol x L(-1) nitrate, sulfate reduction was inhibited for 5, 9, 20, and over 35 days, respectively. With the addition of 0.6, 0.9, 1.4, 2.6 and 4.6 mmol x L(-1) of nitrite, the inhibitory period lasted 3, 12, 22, and over 39 days, respectively. The SRP enrichment culture could dissimilatorily reduce nitrate to ammonium. When sulfate, nitrate and nitrite coexisted, nitrate or nitrite was preferentially used over sulfate as electron acceptor by the enriched SRP. This competitive use of electron acceptor and the strong inhibitory effect of nitrite possibly accounted for the suppression of nitrate and nitrite on the sulfate-reducing activity of the enriched SRP cultures from offshore oil reservoir at Bohai Bay. PMID:25509091

  8. Analysis of denitrifier community in a bioaugmented sequencing batch reactor for the treatment of coking wastewater containing pyridine and quinoline

    Bai, Yaohui; Xing, Rui; Wen, Donghui; Tang, Xiaoyan [Peking Univ., Beijing (CN). Key Lab. of Water and Sediment Sciences (Ministry of Education); Sun, Qinghua [Peking Univ., Beijing (CN). Key Lab. of Water and Sediment Sciences (Ministry of Education); Chinese Center for Disease Control and Prevention, Beijing (China). Inst. of Environmental Health and Related Product Safety

    2011-05-15

    The denitrifier community and associated nitrate and nitrite reduction in the bioaugmented and general sequencing batch reactors (SBRs) during the treatment of coking wastewater containing pyridine and quinoline were investigated. The efficiency and stability of nitrate and nitrite reduction in SBR was considerably improved after inoculation with four pyridine- or quinoline-degrading bacterial strains (including three denitrifying strains). Terminal restriction fragment length polymorphism (T-RFLP) based on the nosZ gene revealed that the structures of the denitrifier communities in bioaugmented and non-bioaugmented reactors were distinct and varied during the course of the experiment. Bioaugmentation protected indigenous denitrifiers from disruptions caused by pyridine and quinoline. Clone library analysis showed that one of the added denitrifiers comprised approximately 6% of the denitrifier population in the bioaugmented sludge. (orig.)

  9. Draft Genome Sequence of Marinobacter sp. Strain P4B1, an Electrogenic Perchlorate-Reducing Strain Isolated from a Long-Term Mixed Enrichment Culture of Marine Bacteria

    Stepanov, Victor G.; Xiao, Yeyuan; Lopez, April J.

    2016-01-01

    The perchlorate-reducing strain Marinobacter sp. strain P4B1 was isolated from a long-term perchlorate-degrading enrichment culture seeded with marine sediment. The draft genome of Marinobacter sp. P4B1 is comprised of the bacterial chromosome (3.60 Mbp, G+C 58.51%, 3,269 predicted genes) and its associated plasmid pMARS01 (0.14 Mbp, G+C 52.95%, 165 predicted genes). PMID:26798109

  10. NirS-containing denitrifier communities in the water column and sediment of the Baltic Sea

    Jürgens, K.; Köster, M. (Miriam); Braker, G.; Wardenga, R.; Hannig, M.; Falk, S.; Gliesche, C.

    2006-01-01

    The aim of this study was to compare structural differences in the denitrifying microbial communities along the environmental gradients observed in the water column and coastal sediments of the Baltic Sea. To link community structure and environmental gradients, denitrifier communities were analyzed by terminal restriction fragment length polymorphism (T-RFLP) based on nirS as a functional marker gene for denitrification. NirS-type denitrifier community composition was further evaluated by ph...

  11. nirS-containing denitrifier communities in the water column and sediment of the Baltic Sea

    Falk, S.; Hannig, M.; Gliesche, C.; Wardenga, R.; Köster, M. (Miriam); Jürgens, K.; Braker, G.

    2007-01-01

    The aim of this study was to compare structural differences in the nirS-type denitrifying microbial communities along the environmental gradients observed in the water column and coastal sediments of the Baltic Sea. To link community structure and environmental gradients, denitrifier communities were analyzed by terminal restriction fragment length polymorphism (T-RFLP) based on nirS as a functional marker gene for denitrification. nirS-type denitrifier community composition...

  12. Denitrifying bacteria from the terrestrial subsurface exposed to mixed waste contamination

    Green, Stefan [Florida State University; Prakash, Om [Florida State University; Gihring, Thomas [Florida State University; Akob, Denise M. [Florida State University; Jasrotia, Puja [Florida State University; Jardine, Philip M [ORNL; Watson, David B [ORNL; Brown, Steven D [ORNL; Palumbo, Anthony Vito [ORNL; Kostka, Joel [Florida State University

    2010-01-01

    In terrestrial subsurface environments where nitrate is a critical groundwater contaminant, few cultivated representatives are available with which to verify the metabolism of organisms that catalyze denitrification. In this study, five species of denitrifying bacteria from three phyla were isolated from subsurface sediments exposed to metal radionuclide and nitrate contamination as part of the U.S. Department of Energy s Oak Ridge Integrated Field Research Challenge (OR-IFRC). Isolates belonged to the genera Afipia and Hyphomicrobium (Alphaproteobacteria), Rhodanobacter (Gammaproteobacteria), Intrasporangium (Actinobacteria) and Bacillus (Firmicutes). Isolates from the phylum Proteobacteria were confirmed as complete denitrifiers, whereas the Gram-positive isolates reduced nitrate to nitrous oxide. Ribosomal RNA gene analyses reveal that bacteria from the genus Rhodanobacter comprise a diverse population of circumneutral to moderately acidophilic denitrifiers at the ORIFRC site, with a high relative abundance in areas of the acidic source zone. Rhodanobacter species do not contain a periplasmic nitrite reductase and have not been previously detected in functional gene surveys of denitrifying bacteria at the OR-IFRC site. Sequences of nitrite and nitrous oxide reductase genes were recovered from the isolates and from the terrestrial subsurface by designing primer sets mined from genomic and metagenomic data and from draft genomes of two of the isolates. We demonstrate that a combination of cultivation, genomic and metagenomic data are essential to the in situ characterization of denitrifiers and that current PCR-based approaches are not suitable for deep coverage of denitrifying microorganisms. Our results indicate that the diversity of denitrifiers is significantly underestimated in the terrestrial subsurface.

  13. Denitrifying bacteria from the terrestrial subsurface exposed to mixed waste contamination

    In terrestrial subsurface environments where nitrate is a critical groundwater contaminant, few cultivated representatives are available with which to verify the metabolism of organisms that catalyze denitrification. In this study, five species of denitrifying bacteria from three phyla were isolated from subsurface sediments exposed to metal radionuclide and nitrate contamination as part of the U.S. Department of Energy's Oak Ridge Integrated Field Research Challenge (OR-IFRC). Isolates belonged to the genera Afipia and Hyphomicrobium (Alphaproteobacteria), Rhodanobacter (Gammaproteobacteria), Intrasporangium (Actinobacteria) and Bacillus (Firmicutes). Isolates from the phylum Proteobacteria were confirmed as complete denitrifiers, whereas the Gram-positive isolates reduced nitrate to nitrous oxide. Ribosomal RNA gene analyses reveal that bacteria from the genus Rhodanobacter comprise a diverse population of circumneutral to moderately acidophilic denitrifiers at the ORIFRC site, with a high relative abundance in areas of the acidic source zone. Rhodanobacter species do not contain a periplasmic nitrite reductase and have not been previously detected in functional gene surveys of denitrifying bacteria at the OR-IFRC site. Sequences of nitrite and nitrous oxide reductase genes were recovered from the isolates and from the terrestrial subsurface by designing primer sets mined from genomic and metagenomic data and from draft genomes of two of the isolates. We demonstrate that a combination of cultivation, genomic and metagenomic data are essential to the in situ characterization of denitrifiers and that current PCR-based approaches are not suitable for deep coverage of denitrifying microorganisms. Our results indicate that the diversity of denitrifiers is significantly underestimated in the terrestrial subsurface.

  14. Effect of pH on the denitrifying enzyme activity in pasture soils in relation to the intrinsic differences in denitrifier communities

    Čuhel, Jiří; Šimek, Miloslav

    2011-01-01

    Roč. 56, č. 3 (2011), s. 230-235. ISSN 0015-5632 R&D Projects: GA MŠk LC06066; GA AV ČR IAA600660605 Institutional research plan: CEZ:AV0Z60660521 Keywords : pH * denitrifying enzyme activity * pasture soils Subject RIV: EH - Ecology, Behaviour Impact factor: 0.677, year: 2011

  15. Anaerobic degradation of toluene by a denitrifying bacterium.

    Evans, P J; Mang, D T; Kim, K S; Young, L Y

    1991-01-01

    A denitrifying bacterium, designated strain T1, that grew with toluene as the sole source of carbon under anaerobic conditions was isolated. The type of agar used in solid media and the toxicity of toluene were determinative factors in the successful isolation of strain T1. Greater than 50% of the toluene carbon was oxidized to CO2, and 29% was assimilated into biomass. The oxidation of toluene to CO2 was stoichiometrically coupled to nitrate reduction and denitrification. Strain T1 was tolerant of and grew on 3 mM toluene after a lag phase. The rate of toluene degradation was 1.8 mumol min-1 liter-1 (56 nmol min-1 mg of protein-1) in a cell suspension. Strain T1 was distinct from other bacteria that oxidize toluene anaerobically, but it may utilize a similar biochemical pathway of oxidation. In addition, o-xylene was transformed to a metabolite in the presence of toluene but did not serve as the sole source of carbon for growth of strain T1. This transformation was dependent on the degradation of toluene. Images PMID:2059037

  16. Seasonal Patterns in Microbial Community Composition in Denitrifying Bioreactors Treating Subsurface Agricultural Drainage.

    Porter, Matthew D; Andrus, J Malia; Bartolerio, Nicholas A; Rodriguez, Luis F; Zhang, Yuanhui; Zilles, Julie L; Kent, Angela D

    2015-10-01

    Denitrifying bioreactors, consisting of water flow control structures and a woodchip-filled trench, are a promising approach for removing nitrate from agricultural subsurface or tile drainage systems. To better understand the seasonal dynamics and the ecological drivers of the microbial communities responsible for denitrification in these bioreactors, we employed microbial community "fingerprinting" techniques in a time-series examination of three denitrifying bioreactors over 2 years, looking at bacteria, fungi, and the denitrifier functional group responsible for the final step of complete denitrification. Our analysis revealed that microbial community composition responds to depth and seasonal variation in moisture content and inundation of the bioreactor media, as well as temperature. Using a geostatistical analysis approach, we observed recurring temporal patterns in bacterial and denitrifying bacterial community composition in these bioreactors, consistent with annual cycling. The fungal communities were more stable, having longer temporal autocorrelations, and did not show significant annual cycling. These results suggest a recurring seasonal cycle in the denitrifying bioreactor microbial community, likely due to seasonal variation in moisture content. PMID:25910602

  17. Impact of Land Use Management and Soil Properties on Denitrifier Communities of Namibian Savannas.

    Braker, Gesche; Matthies, Diethart; Hannig, Michael; Brandt, Franziska Barbara; Brenzinger, Kristof; Gröngröft, Alexander

    2015-11-01

    We studied potential denitrification activity and the underlying denitrifier communities in soils from a semiarid savanna ecosystem of the Kavango region in NE Namibia to help in predicting future changes in N(2)O emissions due to continuing changes of land use in this region. Soil type and land use (pristine, fallow, and cultivated soils) influenced physicochemical characteristics of the soils that are relevant to denitrification activity and N(2)O fluxes from soils and affected potential denitrification activity. Potential denitrification activity was assessed by using the denitrifier enzyme activity (DEA) assay as a proxy for denitrification activity in the soil. Soil type and land use influenced C and N contents of the soils. Pristine soils that had never been cultivated had a particularly high C content. Cultivation reduced soil C content and the abundance of denitrifiers and changed the composition of the denitrifier communities. DEA was strongly and positively correlated with soil C content and was higher in pristine than in fallow or recently cultivated soils. Soil type and the composition of both the nirK- and nirS-type denitrifier communities also influenced DEA. In contrast, other soil characteristics like N content, C:N ratio, and pH did not predict DEA. These findings suggest that due to greater availability of soil organic matter, and hence a more effective N cycling, the natural semiarid grasslands emit more N(2)O than managed lands in Namibia. PMID:25977143

  18. Uranium enrichment

    This paper reports that in 1990 the Department of Energy began a two-year project to illustrate the technical and economic feasibility of a new uranium enrichment technology-the atomic vapor laser isotope separation (AVLIS) process. GAO believes that completing the AVLIS demonstration project will provide valuable information about the technical viability and cost of building an AVLIS plant and will keep future plant construction options open. However, Congress should be aware that DOE still needs to adequately demonstrate AVLIS with full-scale equipment and develop convincing cost projects. Program activities, such as the plant-licensing process, that must be completed before a plant is built, could take many years. Further, an updated and expanded uranium enrichment analysis will be needed before any decision is made about building an AVLIS plant. GAO, which has long supported legislation that would restructure DOE's uranium enrichment program as a government corporation, encourages DOE's goal of transferring AVLIS to the corporation. This could reduce the government's financial risk and help ensure that the decision to build an AVLIS plant is based on commercial concerns. DOE, however, has no alternative plans should the government corporation not be formed. Further, by curtailing a planned public access program, which would have given private firms an opportunity to learn about the technology during the demonstration project, DOE may limit its ability to transfer AVLIS to the private sector

  19. Uranium enrichment

    This chapter discusses the development of uranium enrichment processes. In the introduction there is a brief history of uranium enrichment, followed by a summary of the criteria used for the assessment of an isotope separation process, e.g. the separation factor, separative power, and the power consumption of a separating element. This is followed by a discussion of the two main processes used, i.e. gaseous diffusion and centrifugation. The reason for the change from diffusion to centrifugation in the UK, mainly on power costs, is discussed. The development potential of centrifuges is also assessed. Other processes which have been developed up to pilot stage are described, e.g. the Becker jet nozzle and the South African process. This is followed by a description of some plasma-based methods. The next topic is concerned with chemical exchange methods and an attempt is made to assess their potential in the enrichment scene from published information. This chapter concludes with a discussion of the advanced laser isotope-separation methods. The two approaches, i.e. the atomic and the molecular routes are discussed again using published information. This information is insufficient to give a complete assessment of the methods, especially the molecular route, but is enough to give indications of their potential

  20. Differential Isotopic Fractionation during Cr(VI) Reduction by an Aquifer-Derived Bacterium under Aerobic versus Denitrifying Conditions

    Han, R.; Qin, L.; Brown, S. T.; Christensen, J. N.; Beller, H. R.

    2012-01-27

    We studied Cr isotopic fractionation during Cr(VI) reduction by Pseudomonas stutzeri strain RCH2. Finally, despite the fact that strain RCH2 reduces Cr(VI) cometabolically under both aerobic and denitrifying conditions and at similar specific rates, fractionation was markedly different under these two conditions (ε was ~2‰ aerobically and ~0.4‰ under denitrifying conditions).

  1. Porównawcze badania nad występowaniem bakterii w hydroponicznych uprawach sałaty (Lactuca sativa L. z azotanowaą i amonową formą azotu w pożywce [Comparative studies on the occurrence of bacteria in hydroponic cultures of lettuce (Lactuca sativa L. enriched with nitrate or ammonia forms of nitrogen in the nutrient medium

    Z. Kobierzyńska-Gołąb

    2015-06-01

    Full Text Available The experiment demonstrated the greatest number of bacteria on the surface of roots immersed in nutrient solution, next on the epidermis of the parts of roots remaining in the air space above the nutrient solution and on the roots growing in the seed-bed; a smaller number in the seed-bed itself and the smallest number in the nutrient solution. The population of bacteria consisted of up to 90% short rod of bacilli. The total number of bacteria in hydroponic culture with ammonium was higher than in that with nitrate. As a rule the bacteria belonging to the separate physiological groups, participating in nitrogen metabolism (ammonifying, proteolytic, proteinizing, denitrifying and oligonitrophilic bacteria appeared to be more numerus in the culture with ammonium than in that with nitrate. The growth of plants in hydroponic culture with ammonium was weaker than in that with nitrate.

  2. Effect of electro-stimulation on activity of heterotrophic denitrifying bacteria and denitrification performance.

    Liu, Hengyuan; Tong, Shuang; Chen, Nan; Liu, Ying; Feng, Chuanping; Hu, Qili

    2015-11-01

    The effects of electro-stimulation on heterotrophic denitrifying bacterial activity and nitrate removal were investigated using a bench-scale bio-electrochemical reactor in this study. Results showed that the maximum nitrate removal efficiency was 100% at the optimum current density of 200mA/m(2), at which low nitrite production and high ATP aggregate level were obtained. The activity of denitrifying bacteria was highest at the range densities of 200-250mA/m(2), although the terminative pH increased to 8.62 at 200mA/m(2) and 9.63 at 250mA/m(2). This demonstrates that suitable current densities could improve the activity of denitrifying bacteria. Therefore, this study provides a number of useful information to improve the bio-electrochemical reactor designs and promote the removal efficiency of pollutants. PMID:26231132

  3. Nitrification and denitrifying phosphorus removal in an upright continuous flow reactor.

    Reza, Maryam; Alvarez Cuenca, Manuel

    2016-01-01

    Simultaneous nitrification and denitrifying phosphorus removal was achieved in a single-sludge continuous flow bioreactor. The upright bioreactor was aligned with a biomass fermenter (BF) and operated continuously for over 350 days. This study revealed that unknown bacteria of the Saprospiraceae class may have been responsible for the successful nutrient removal in this bioreactor. The successive anoxic-aerobic stages of the bioreactor with upright alignment along with a 60 L BF created a unique ecosystem for the growth of nitrifier, denitrifiers, phosphorus accumulating organisms and denitrifying phosphorus accumulating organisms. Furthermore, total nitrogen to chemical oxygen demand (COD) ratio and total phosphorus to COD ratio of 0.6 and 0.034, respectively, confirmed the comparative advantages of this advanced nutrient removal process relative to both sequencing batch reactors and activated sludge processes. The process yielded 95% nitrogen removal and over 90% phosphorus removal efficiencies. PMID:27148710

  4. Concurrent Activity of Anammox and Denitrifying Bacteria in the Black Sea

    JohnB.Kirkpatrick

    2012-07-01

    Full Text Available After the discovery of ANaerobic AMMonium OXidation (anammox in the Black Sea in 2003, the role of heterotrophic denitrification as the main marine pathway for fixed N loss was questioned. A 3 part, 15 month time series investigating Black Sea nitrite reductase (nirS mRNA transcripts at a single location was conducted in order to better understand the activity of anammox and denitrifying bacteria. Here we show that both of these groups were active, as well as being concurrent in the lower suboxic zone over this time span. Their distributions, however, were fundamentally different; denitrification expression was much more variable. Depth profiles covering the suboxic zone showed that the four groups of anammox-type sequences were expressed consistently in the lower suboxic zone, and were consistent with anammox 16S rDNA gene profiles. By contrast, denitrifier-type nirS sequence groups were mixed; half of the groups exhibited consistent expression in the lower suboxic zone, while others varied between season and depth. Co-occurrence of both anammox and denitrifier expression was common and ongoing. Both types of transcripts were also found in samples with low concentrations of sulfide ( >2 μM. 6 major groups of denitrifier-type nirS transcripts were identified, making these transcripts more diverse than previous DNA clone libraries. Several groups of denitrifier-type nirS transcripts were closely related to sequences from the Baltic Sea. An increase in denitrifier-type nirS transcript diversity and depth range in October 2007 corresponded to an increase in mixed layer net community productivity as measured by O2/Ar gas ratios, as well as to an increase in N2 concentrations in the upper suboxic zone. Taken together, the variations in expression patterns between anammox and denitrification provide one explanation as to how near instantaneous rate measurements, such as isotope spike experiments, may regularly detect anammox activity but underreport denitrification.

  5. Mapping spatial patterns of denitrifiers for bridging community ecology and microbial processes along environmental gradients

    Bru, D.; Cuhel, J.; Saby, N.; Cheneby, D.; Chronokova, A.; Arrouays, D.; Martin-Laurent, F.; Simek, M.

    2010-12-01

    While there is ample evidence that microbial processes can exhibit large variations at a field scale, very little is known about the spatial distribution of the communities mediating these processes. To explore spatial patterns of size and activity of the denitrifying community, a functional guild involved in N-cycling, in a grassland field subjected to different cattle grazing regimes.We used geostatistical modeling to map the distribution of size and activity of the denitrifier community in the pasture. Size of the denitrifier community was estimated by PCR quantification of the denitrification gene copy numbers while its activity was estimated by measuring potential denitrification activity and potential N2O emissions. Non-random distribution patterns of the size and of the activity of the denitrifier community were observed with a field-scale spatial dependence. The soil properties, which were strongly affected by presence of cattle, imposed significant control on potential denitrification activity, potential N2O production but not on the size of the denitrifier community. The relative abundance of bacteria possessing the nosZ gene encoding the N2O reductase within the total bacterial community was a strong predictor of the N2O/N2 ratio. Our results clearly indicated that patterns of distribution of the abundance of denitrifiers can be modelled at a field scale. Characterization of such pattern at a field-scale constitutes the first step in modelling distribution of functional bacterial communities at a scale compatible with land management strategies. The absolute abundance of most denitrification genes was not correlated with potential denitrification activity or potential N2O production. However, the relative abundance of bacteria possessing the nosZ gene in the total bacterial community was a strong predictor of the N2O/(N2+N2O) ratio, suggesting a relationship between ecosystem processes and bacterial community composition.

  6. Effect of pyrene on denitrification activity and abundance and composition of denitrifying community in an agricultural soil

    Toxicity of pyrene on the denitrifiers was studied by spiking an agricultural soil with pyrene to a series of concentrations (0-500 mg kg-1) followed by dose-response and dynamic incubation experiments. Results showed a positive correlation between potential denitrification activity and copy numbers of denitrifying functional genes (nirK, nirS and nosZ), and were both negatively correlated with pyrene concentrations. Based on the comparison of EC50 values, denitrifiers harboring nirK, nirS or nosZ gene were more sensitive than denitrification activity, and denitrifiers harboring nirS gene were more sensitive than that harboring nirK or nosZ genes. Seven days after spiking with EC50 concentration of pyrene, denitrifiers diversity decreased and community composition changed in comparison with the control. Phylogenetic analyses of three genes showed that the addition of pyrene increased the proportion of Bradyrhizobiaceae, Rhodospirillales, Burkholderiales and Pseudomonadales. Some species belonging to these groups were reported to be able to degrade PAHs. - Highlights: → Toxicity of pyrene on the denitrifiers was studied by spiking an agricultural soil with pyrene. → PDA was positively correlated with the abundance of denitrifiers harboring nirK, nirS or nosZ gene. → Both PDA and the abundance of denitrifiers were negatively correlated with pyrene concentrations. → Denitrifiers harboring nirk, nirS or nosZ gene are more sensitive to pyrene than PDA in soils. - Denitrifiers harboring nirK, nirS or nosZ gene are more sensitive to pyrene contamination than potential denitrification activity in soils.

  7. Effect of pyrene on denitrification activity and abundance and composition of denitrifying community in an agricultural soil

    Guo Guangxia; Deng Huan; Qiao Min; Mu Yujing [State Key Lab of Regional and Urban Ecology, Research Center for Eco-Environmental Sciences, Chinese Academy of Sciences, Beijing 100085 (China); Zhu Yongguan, E-mail: ygzhu@rcees.ac.cn [State Key Lab of Regional and Urban Ecology, Research Center for Eco-Environmental Sciences, Chinese Academy of Sciences, Beijing 100085 (China); Key Lab of Urban Environment and Health, Institute of Urban Environment, Chinese Academy of Sciences, Xiamen 361021 (China)

    2011-07-15

    Toxicity of pyrene on the denitrifiers was studied by spiking an agricultural soil with pyrene to a series of concentrations (0-500 mg kg{sup -1}) followed by dose-response and dynamic incubation experiments. Results showed a positive correlation between potential denitrification activity and copy numbers of denitrifying functional genes (nirK, nirS and nosZ), and were both negatively correlated with pyrene concentrations. Based on the comparison of EC{sub 50} values, denitrifiers harboring nirK, nirS or nosZ gene were more sensitive than denitrification activity, and denitrifiers harboring nirS gene were more sensitive than that harboring nirK or nosZ genes. Seven days after spiking with EC{sub 50} concentration of pyrene, denitrifiers diversity decreased and community composition changed in comparison with the control. Phylogenetic analyses of three genes showed that the addition of pyrene increased the proportion of Bradyrhizobiaceae, Rhodospirillales, Burkholderiales and Pseudomonadales. Some species belonging to these groups were reported to be able to degrade PAHs. - Highlights: > Toxicity of pyrene on the denitrifiers was studied by spiking an agricultural soil with pyrene. > PDA was positively correlated with the abundance of denitrifiers harboring nirK, nirS or nosZ gene. > Both PDA and the abundance of denitrifiers were negatively correlated with pyrene concentrations. > Denitrifiers harboring nirk, nirS or nosZ gene are more sensitive to pyrene than PDA in soils. - Denitrifiers harboring nirK, nirS or nosZ gene are more sensitive to pyrene contamination than potential denitrification activity in soils.

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

    Vieira-Dalodé, G.; Madodé, Y.E.; Hounhouigan, J.; Jespersen, Lene; Jakobsen, Mogens

    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...

  9. 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.

    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. PMID:26408128

  10. Pyruvic oxime nitrification and copper and nickel resistance by a Cupriavidus pauculus, an active heterotrophic nitrifier-denitrifier.

    Ramirez, Miguel; Obrzydowski, Jennifer; Ayers, Mary; Virparia, Sonia; Wang, Meijing; Stefan, Kurtis; Linchangco, Richard; Castignetti, Domenic

    2014-01-01

    Heterotrophic nitrifiers synthesize nitrogenous gasses when nitrifying ammonium ion. A Cupriavidus pauculus, previously thought an Alcaligenes sp. and noted as an active heterotrophic nitrifier-denitrifier, was examined for its ability to produce nitrogen gas (N2) and nitrous oxide (N2O) while heterotrophically nitrifying the organic substrate pyruvic oxime [CH3-C(NOH)-COOH]. Neither N2 nor N2O were produced. Nucleotide and phylogenetic analyses indicated that the organism is a member of a genus (Cupriavidus) known for its resistance to metals and its metabolism of xenobiotics. The microbe (a Cupriavidus pauculus designated as C. pauculus strain UM1) was examined for its ability to perform heterotrophic nitrification in the presence of Cu(2+) and Ni(2+) and to metabolize the xenobiotic phenol. The bacterium heterotrophically nitrified well when either 1?mM Cu(2+) or 0.5?mM Ni(2+) was present in either enriched or minimal medium. The organism also used phenol as a sole carbon source in either the presence or absence of 1?mM Cu(2+) or 0.5?mM Ni(2+). The ability of this isolate to perform a number of different metabolisms, its noteworthy resistance to copper and nickel, and its potential use as a bioremediation agent are discussed. PMID:25580463

  11. Environmental evaluation of coexistence of denitrifying anaerobic methane-oxidizing archaea and bacteria in a paddy field.

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

    2016-01-01

    The nitrate-dependent denitrifying anaerobic methane oxidation (DAMO) process, which is metabolized together by anaerobic methanotrophic archaea and NC10 phylum bacteria, is expected to be important for the global carbon and nitrogen cycles. However, there are little studies about the existence of this process and the functional microbes in environments. Therefore, the coexistence of DAMO archaea and bacteria in a paddy field was evaluated in this study. Next-generation sequencing showed that the two orders, Methanosarcinales and Nitrospirales, to which DAMO archaea and DAMO bacteria belong, were detected in the four soil samples. Then the in vitro experiments demonstrated both of nitrite- and nitrate-dependent DAMO activities, which confirmed the coexistence of DAMO archaea and DAMO bacteria. It was the first report about the coexistence of DAMO archaea and bacteria in a paddy field. Furthermore, anammox bacteria were detected in two of the four samples. The in vitro experiments did not show anammox activity in the initial period but showed low anammox activity after 20 days' enrichment. These results implicated that anammox bacteria may coexist with DAMO microorganisms in this field, but at a very low percentage. PMID:26394860

  12. Nitrogen Removal from Micro-Polluted Reservoir Water by Indigenous Aerobic Denitrifiers

    Ting-Lin Huang

    2015-04-01

    Full Text Available Treatment of micro-polluted source water is receiving increasing attention because of environmental awareness on a global level. We isolated and identified aerobic denitrifying bacteria Zoogloea sp. N299, Acinetobacter sp. G107, and Acinetobacter sp. 81Y and used these to remediate samples of their native source water. We first domesticated the isolated strains in the source water, and the 48-h nitrate removal rates of strains N299, G107, and 81Y reached 33.69%, 28.28%, and 22.86%, respectively, with no nitrite accumulation. We then conducted a source-water remediation experiment and cultured the domesticated strains (each at a dry cell weight concentration of 0.4 ppm together in a sample of source water at 20–26 °C and a dissolved oxygen concentration of 3–7 mg/L for 60 days. The nitrate concentration of the system decreased from 1.57 ± 0.02 to 0.42 ± 0.01 mg/L and that of a control system decreased from 1.63 ± 0.02 to 1.30 ± 0.01 mg/L, each with no nitrite accumulation. Total nitrogen of the bacterial system changed from 2.31 ± 0.12 to 1.09 ± 0.01 mg/L, while that of the control system changed from 2.51 ± 0.13 to 1.72 ± 0.06 mg/L. The densities of aerobic denitrification bacteria in the experimental and control systems ranged from 2.8 × 104 to 2 × 107 cfu/mL and from 7.75 × 103 to 5.5 × 105 cfu/mL, respectively. The permanganate index in the experimental and control systems decreased from 5.94 ± 0.12 to 3.10 ± 0.08 mg/L and from 6.02 ± 0.13 to 3.61 ± 0.11 mg/L, respectively, over the course of the experiment. Next, we supplemented samples of the experimental and control systems with additional bacteria or additional source water and cultivated the systems for another 35 days. The additional bacteria did little to improve the water quality. The additional source water provided supplemental carbon and brought the nitrate removal rate in the experimental system to 16.97%, while that in the control system reached only 3.01%, with no nitrite accumulation in either system. Our results show that aerobic denitrifying bacteria remain highly active after domestication and demonstrate the applicability of such organisms in the bioremediation of oligotrophic ecosystems.

  13. Self-protected nitrate reducing culture for intrinsic repair of concrete cracks.

    Erşan, Yusuf Ç; Gruyaert, Elke; Louis, Ghislain; Lors, Christine; De Belie, Nele; Boon, Nico

    2015-01-01

    Attentive monitoring and regular repair of concrete cracks are necessary to avoid further durability problems. As an alternative to current maintenance methods, intrinsic repair systems which enable self-healing of cracks have been investigated. Exploiting microbial induced CaCO3 precipitation (MICP) using (protected) axenic cultures is one of the proposed methods. Yet, only a few of the suggested healing agents were economically feasible for in situ application. This study presents a [Formula: see text] reducing self-protected enrichment culture as a self-healing additive for concrete. Concrete admixtures Ca(NO3)2 and Ca(HCOO)2 were used as nutrients. The enrichment culture, grown as granules (0.5-2 mm) consisting of 70% biomass and 30% inorganic salts were added into mortar without any additional protection. Upon 28 days curing, mortar specimens were subjected to direct tensile load and multiple cracks (0.1-0.6 mm) were achieved. Cracked specimens were immersed in water for 28 days and effective crack closure up to 0.5 mm crack width was achieved through calcite precipitation. Microbial activity during crack healing was monitored through weekly NOx analysis which revealed that 92 ± 2% of the available [Formula: see text] was consumed. Another set of specimens were cracked after 6 months curing, thus the effect of curing time on healing efficiency was investigated, and mineral formation at the inner crack surfaces was observed, resulting in 70% less capillary water absorption compared to healed control specimens. In conclusion, enriched mixed denitrifying cultures structured in self-protecting granules are very promising strategies to enhance microbial self-healing. PMID:26583015

  14. Impact of Flood Spates on Denitrifying Bacteria in Low Order Streams

    Herrman, K.; Stokdyk, J.

    2011-12-01

    The impact of flood events on channel design, macroinvertebrates, and periphyton in stream ecosystems has been well studied. Little is known, however, about how flood spates affect microorganisms found in stream sediments. Denitrifying bacteria are beneficial organisms because they convert nitrates to nitrogen gas. Providing data that describes the impact of flood events on denitrifiers and the time required after the disturbance for the bacteria to recover are crucial in understanding nitrogen dynamics in stream ecosystems. Three low order streams in central Wisconsin, USA are being monitored during several flood spates during July and August of 2011. Discharge is being continuously monitored in all three streams and sediments are being collected before and after several flood events for laboratory assays. Specifically, sediments are being processed for denitrification rates using the acetylene inhibition technique, microbial biomass carbon using chloroform fumigation, and the quantification of denitrifying bacteria (i.e., nirS, nirK, and nosZ genes) using real-time quantitative PCR. Preliminary data show that within 36 hours after a 90 mm rain event, microbial biomass carbon in all three streams (580 μg C g sediment-1) significantly increased (F1,23 = 650 ± 140; p bacteria in stream sediments. Denitrification rates and quantification of denitrifying bacteria still need to be analyzed to determine if these specific bacteria follow a similar pattern or if the bacterial recolonization of stream sediments follows a unique pattern.

  15. Cattle activities affect abundance and activity of nitrifying and denitrifying microbial communities in upland soil

    Chroňáková, Alica; Radl, V.; Čuhel, Jiří; Gattinger, A.; Šimek, Miloslav; Elhottová, Dana; Schloter, M.

    Uppsala : Swedish University of Agriculture Sciences, 2007. [Achievements of COST 856. Denitrification and related aspects. Final meeting of the ESF COST Action 856 /14./. 05.12.2007-08.12.2007, Uppsala] Institutional research plan: CEZ:AV0Z60660521 Keywords : cattle activities * nitrifying and denitrifying microbial communities * upland soil Subject RIV: EH - Ecology, Behaviour

  16. Post cold-storage conditioning time affects soil denitrifying enzyme activity

    Chirinda, Ngoni; Olesen, Jørgen E; Porter, John R.

    2011-01-01

    Soil denitrifying enzyme activity (DEA) is often assessed after cold storage. Previous studies using the short-term acetylene inhibition method have not considered conditioning time (post-cold-storage warm-up time prior to soil analysis) as a factor influencing results. We observed fluctuations i...

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

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

  18. Mapping spatial patterns of denitrifiers for bridging community ecology and microbial processes along environmental gradients

    Philippot, L.; Čuhel, Jiří; Saby, N.P.A.; Chéneby, D.; Chroňáková, Alica; Bru, D.; Arrouays, D.; Martin-Laurent, F.; Šimek, Miloslav

    Uppsala : Swedish University of Agricultural Sciences, 2009. s. 110. [BAGECO 10. Bacterial Genetics and Ecology - Coexisting on a Changing Planet. 15.06.2009-19.06.2009, Uppsala] Institutional research plan: CEZ:AV0Z60660521 Keywords : spatial patterns * denitrifiers * microbial processes Subject RIV: EH - Ecology, Behaviour

  19. cultural

    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.

  20. RELATIONSHIP BETWEEN THE CONCENTRATION OF DENITRIFIERS AND PSEUDOMONAS SPP. IN SOILS: IMPLICATIONS FOR BTX BIOREMEDIATION (R823420)

    Aquifer microcosms were used to investigate the effect of stimulating denitrification on microbial population shifts and BTX degradation potential. Selective pressurefor facultative denitrifiers was applied to a treatment set by feeding acetate and nitrate, and cycling electr...

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

    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.

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

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

    2016-01-01

    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. PMID:26771607

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

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

    2016-01-01

    Anaerobic digestion using mixed-culture with broader choice of pretreatments for hydrogen (H?) 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 H? 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 H? consuming bacteria and produced twice as much as H? 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 H? production as response parameter. The maximum H? 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 H? 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 H? production. PMID:26771607

  4. 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

    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.

  5. Simultaneous removal of sulfide, nitrate and acetate under denitrifying sulfide removal condition: Modeling and experimental validation

    Xu, Xijun; Chen, Chuan; Wang, Aijie; Guo, Wanqian; Zhou, Xu [State Key Laboratory of Urban Water Resource and Environment, Harbin Institute of Technology, Harbin 150090 (China); Lee, Duu-Jong, E-mail: djlee@ntu.edu.tw [State Key Laboratory of Urban Water Resource and Environment, Harbin Institute of Technology, Harbin 150090 (China); Department of Chemical Engineering, National Taiwan University, Taipei 106, Taiwan (China); Department of Chemical Engineering, National Taiwan University of Science and Technology, Taipei 106, Taiwan (China); Ren, Nanqi, E-mail: rnq@hit.edu.cn [State Key Laboratory of Urban Water Resource and Environment, Harbin Institute of Technology, Harbin 150090 (China); Chang, Jo-Shu [Research Center for Energy Technology and Strategy, National Cheng Kung University, Tainan, Taiwan (China)

    2014-01-15

    Graphical abstract: Model evaluation applied to case study 1: (A-G) S{sup 2−}, NO{sub 3}{sup −}-N, NO{sub 2}{sup −}-N, and Ac{sup −}-C profiles under initial sulfide concentrations of 156.2 (A), 539 (B), 964 (C), 1490 (D), 342.7 (E), 718 (F), and 1140.7 (G) mg L{sup −1}. The solid line represents simulated result and scatter represents experimental result. -- Highlights: • This work developed a mathematical model for DSR process. • Kinetics of sulfur–nitrogen–carbon and interactions between denitrifiers were studied. • Kinetic parameters of the model were estimated via data fitting. • The model described kinetic behaviors of DSR processes over wide parametric range. -- Abstract: Simultaneous removal of sulfide (S{sup 2−}), nitrate (NO{sub 3}{sup −}) and acetate (Ac{sup −}) under denitrifying sulfide removal process (DSR) is a novel biological wastewater treatment process. This work developed a mathematical model to describe the kinetic behavior of sulfur–nitrogen–carbon and interactions between autotrophic denitrifiers and heterotrophic denitrifiers. The kinetic parameters of the model were estimated via data fitting considering the effects of initial S{sup 2−} concentration, S{sup 2−}/NO{sub 3}{sup −}-N ratio and Ac{sup −}-C/NO{sub 3}{sup −}-N ratio. Simulation supported that the heterotrophic denitratation step (NO{sub 3}{sup −} reduction to NO{sub 2}{sup −}) was inhibited by S{sup 2−} compared with the denitritation step (NO{sub 2}{sup −} reduction to N{sub 2}). Also, the S{sup 2−} oxidation by autotrophic denitrifiers was shown two times lower in rate with NO{sub 2}{sup −} as electron acceptor than that with NO{sub 3}{sup −} as electron acceptor. NO{sub 3}{sup −} reduction by autotrophic denitrifiers occurs 3–10 times slower when S{sup 0} participates as final electron donor compared to the S{sup 2−}-driven pathway. Model simulation on continuous-flow DSR reactor suggested that the adjustment of hydraulic retention time is an efficient way to make the reactor tolerating high S{sup 2−} loadings. The proposed model properly described the kinetic behaviors of DSR processes over wide parametric ranges and which can offer engineers with basis to optimize bioreactor operation to improve the treatment capacity.

  6. Characterization of Denitrifying Phosphorus Removal Microorganisms in a Novel Two-Sludge Process by Combining Chemical with Microbial Analysis

    Haiming Zou; Xiwu Lu; Saad Abualhail

    2014-01-01

    The present work focuses on the investigation of denitrifying phosphorus removal organisms (DPB) in a novel two-sludge denitrifying phosphorus removal process by combining chemical with microbial analysis. When the two-sludge process operated stably over one year, good phosphorus (P) release and P uptake performance of activated sludge samples collected from this process were present in anaerobic and anoxic conditions, respectively, via batch test, showing that the ratio of P release specific...

  7. Relationship between Nitrite Reduction and Active Phosphate Uptake in the Phosphate-Accumulating Denitrifier Pseudomonas sp. Strain JR 12

    Barak, Yoram; van Rijn, Jaap

    2000-01-01

    Phosphate uptake by the phosphate-accumulating denitrifier Pseudomonas sp. JR12 was examined with different combinations of electron and carbon donors and electron acceptors. Phosphate uptake in acetate-supplemented cells took place with either oxygen or nitrate but did not take place when nitrite served as the final electron acceptor. Furthermore, nitrite reduction rates by this denitrifier were shown to be significantly reduced in the presence of phosphate. Phosphate uptake assays in the pr...

  8. Performance of a reactor containing denitrifying immobilized biomass in removing ethanol and aromatic hydrocarbons (BTEX) in a short operating period

    A horizontal-flow anaerobic immobilized biomass reactor (HAIB) containing denitrifying biomass was evaluated with respect to its ability to remove, separately and in a short operating period (30 days), organic matter, nitrate, and the hydrocarbons benzene (41.4 mg L-1), toluene (27.8 mg L-1), ethylbenzene (31.1 mg L-1), o-xylene (28.5 mg L-1), m-xylene (28.4 mg L-1) and p-xylene (32.1 mg L-1). The purified culture, which was grown in the presence of the specific hydrocarbon, was used as the source of cells to be immobilized in the polyurethane foam. After 30 days of operation, the foam was removed and a new immobilized biomass was grown in the presence of another hydrocarbon. The average hydrocarbon removal efficiency attained was 97%. The organic matter, especially ethanol, was removed with an average efficiency of 83% at a mean influent concentration of 1185.0 mg L-1. A concomitant removal of 97% of nitrate was observed for a mean influent concentration of 423.4 mg L-1. The independent removal of each hydrocarbon demonstrated that these contaminants can be biodegraded separately, without the need for a compound to be the primary substrate for the degradation of another. This study proposes the application of the system for treatment of areas contaminated with these compounds, with substitution and formation of a biofilm in a 30-day period

  9. Influence of environmental factors on dissolved nitrate stable isotopes under denitrifying conditions - carbon sources and water isotopes

    Wunderlich, A.; Meckenstock, R.; Einsiedl, F.

    2012-04-01

    Stable isotopes in dissolved nitrate are regularly used to identify sources of nitrate contamination in aquifers and water bodies. A dual isotope plot of 15N and 18O in nitrate can provide good evidence of the origin of such pollution as various sources have different isotopic signatures. Microbial denitrification changes both isotopic values by removing nitrate with lighter isotopes first, thereby increasing δ18O as well as δ15N. This change can distort the determination of sources but also has the potential to be used to identify and quantify microbial denitrification. Previous studies found a wide range of enrichment factors (ɛ) that did not allow conclusions towards the extent of microbial denitrification. However, it was found that during denitrification at each respective field site or laboratory experiment, there was a constant ratio in increase of the values of δ18O in relation to δ15N. That ratio was, however, not constant across field sites and the values published range from below 0.5 to more than 1.0. The reasons for these variations in enrichment factors and relative enrichment of oxygen compared to nitrogen are yet unknown. We conducted microcosm experiments with three different bacterial species to elucidate possible influences of environmental factors on these parameters. As a result we conclude that the type of carbon source available to denitrifying bacteria can play a role in the value of the enrichment factors, but not in the relative enrichment of the two isotopes. Specifically we found that complex hydrocarbons (toluene, benzoate) produce significantly different enrichment factors in nitrate than a simple hydrocarbon substrate (acetate). The relative enrichment of δ18O compared to δ15N was 0.86. We hypothesise that this influence is based on a variation in process kinetics of cross-membrane nitrate transport in relation to intracellular nitrate reduction. The core of the hypothesis is that nitrate transport into the cell becomes rate limiting as a result of a carbon source induced change in cell membrane composition. The apparent kinetic isotope effect observed outside the cell is then changed as transport-related isotope effects dominate the observations. In addition, a possible effect of water δ18O values on the δ18O of dissolved nitrate was researched. Intermediary nitrite is known to exchange oxygen atoms with water; a reverse reaction of the nitrate reducing step could thus influence the oxygen isotope composition of dissolved nitrate without changing the nitrogen isotopic composition in the same way. Such a process was already shown for sulfate reduction. By adding 18O-labelled water to microcosm experiments, we could show that such an exchange exists for selected microorganisms. The environmental implications of this result is discussed.

  10. Denitrification potential under different fertilization regimes is closely coupled with changes in the denitrifying community in a black soil.

    Yin, Chang; Fan, Fenliang; Song, Alin; Cui, Peiyuan; Li, Tingqiang; Liang, Yongchao

    2015-07-01

    Preferable inorganic fertilization over the last decades has led to fertility degradation of black soil in Northeast China. However, how fertilization regimes impact denitrification and its related bacterial community in this soil type is still unclear. Here, taking advantage of a suit of molecular ecological tools in combination of assaying the potential denitrification (DP), we explored the variation of activity, community structure, and abundance of nirS and nirK denitrifiers under four different fertilization regimes, namely no fertilization control (N0M0), organic pig manure (N0M1), inorganic fertilization (N1M0), and combination of inorganic fertilizer and pig manure (N1M1). The results indicated that organic fertilization increased DP, but inorganic fertilization had no impacts. The increase of DP was mirrored by the shift of nirS denitrifiers' community structure but not by that of nirK denitrifiers'. Furthermore, the change of DP coincided with the variation of abundances of both denitrifiers. Shifts of community structure and abundance of nirS and nirK denitrifiers were correlated with the change of soil pH, total nitrogen (TN), organic matter (OM), C:P, total phosphorus (TP), and available phosphorus (Olsen P). Our results suggest that the change of DP under these four fertilization regimes was closely related to the shift of denitrifying bacteria communities resulting from the variation of properties in the black soil tested. PMID:25715781

  11. Development of a Cell Culture Method To Isolate and Enrich Salmonella enterica Serotype Enteritidis from Shell Eggs for Subsequent Detection by Real-Time PCR▿

    Day, J. B.; Basavanna, U.; Sharma, S. K.

    2009-01-01

    Salmonella enterica serotype Enteritidis is a major cause of nontyphoidal salmonellosis from ingestion of contaminated raw or undercooked shell eggs. Current techniques used to identify Salmonella serotype Enteritidis in eggs are extremely laborious and time-consuming. In this study, a novel eukaryotic cell culture system was combined with real-time PCR analysis to rapidly identify Salmonella serotype Enteritidis in raw shell eggs. The system was compared to the standard microbiological metho...

  12. Nitrous oxide reductase (nosZ) gene-specific PCR primers for detection of denitrifiers and three nosZ genes from marine sediments.

    Scala, D J; Kerkhof, L J

    1998-05-01

    Two PCR primer sets for the nitrous oxide reductase gene (nosZ) were developed. The initial primers were based on three sequences in GenBank and used to amplify nosZ from continental shelf sediments and from two denitrifiers in culture, Thiosphaera pantotropha and Pseudomonas denitrificans. Three unique marine sediment nosZ genes were identified and sequenced. The marine nosZ genes were most closely related to the nosZ genes of Paracoccus denitrificans or to Rhizobium meliloti. Alignment of all nosZ sequences currently available (n = 10) facilitated redesign of the PCR primers. Three new primer sets which amplify 1100 bp, 900 bp and 250 bp regions of the nosZ gene were designed and tested. The new primers robustly amplified nosZ fragments from samples in which the initial nosZ primers were only marginally successful. PMID:9595664

  13. Relative rates of nitric oxide and nitrous oxide production by nitrifiers, denitrifiers, and nitrate respirers

    Anderson, I. C.; Levine, J. S.

    1986-01-01

    An account is given of the atmospheric chemical and photochemical effects of biogenic nitric and nitrous oxide emissions. The magnitude of the biogenic emission of NO is noted to remain uncertain. Possible soil sources of NO and N2O encompass nitrification by autotropic and heterotropic nitrifiers, denitrification by nitrifiers and denitrifiers, nitrate respiration by fermenters, and chemodenitrification. Oxygen availability is the primary determinant of these organisms' relative rates of activity. The characteristics of this major influence are presently investigated in light of the effect of oxygen partial pressure on NO and N2O production by a wide variety of common soil-nitrifying, denitrifying, and nitrate-respiring bacteria under laboratory conditions. The results obtained indicate that aerobic soils are primary sources only when there is sufficient moisture to furnish anaerobic microsites for denitrification.

  14. Simultaneous production of acetate and methane from glycerol by selective enrichment of hydrogenotrophic methanogens in extreme-thermophilic (70 °C) mixed culture fermentation

    Highlights: • Simultaneous production of acetate and methane from glycerol was investigated. • Acetate accounted for more than 90% of metabolites in liquid solutions. • The maximum concentration of acetate was above 13 g/L. • 93% of archaea were hydrogenotrophic methanogens. • Thermoanaerobacter was main bacterium and its percentage was 92%. - Abstract: The feasibility of simultaneous production of acetate and methane from glycerol was investigated by selective enrichment of hydrogenotrophic methanogens in an extreme-thermophilic (70 °C) fermentation. Fed-batch experiments showed acetate was produced at the concentration up to 13.0 g/L. A stable operation of the continuous stirred tank reactor (CSTR) was reached within 100 days. Acetate accounted for more than 90 w/w% of metabolites in the fermentation liquid. The yields of methane and acetate were close to the theoretical yields with 0.74–0.80 mol-methane/mol-glycerol and 0.63–0.70 mol-acetate/mol-glycerol. The obtained microbial community was characterized. Hydrogenotrophic methanogens, mainly Methanothermobacter thermautotrophicus formed 93% of the methanogenogenic community. This confirms that a high temperature (70 °C) could effectively select for hydrogenotrophic methanogenic archaea. Thermoanaerobacter spp. was the main bacterium forming 91.5% of the bacterial population. This work demonstrated the conversion of the byproduct of biodiesel production, glycerol, to acetate as a chemical and biogas for energy generation

  15. Insight into the short- and long-term effects of Cu(II) on denitrifying biogranules.

    Chen, Hui; Chen, Qian-Qian; Jiang, Xiao-Yan; Hu, Hai-Yan; Shi, Man-Ling; Jin, Ren-Cun

    2016-03-01

    This study aimed to investigate the short- and long-term effects of Cu(2+) on the activity and performance of denitrifying bacteria. The short-term effects of various concentrations of Cu(2+) on the denitrifying bacteria were evaluated using batch assays. The specific denitrifying activity (SDA) decreased from 14.3 ± 2.2 (without Cu(2+)) to 6.1 ± 0.1 mg N h(-1)g(-1) VSS (100 mg Cu(2+)L(-1)) when Cu(2+) increased from 0 to 100 mg L(-1) with an increment of 10 mg Cu(2+)L(-1). A non-competitive inhibition model was used to calculate the 50% inhibition concentration (IC50) of Cu(2+) on denitrifying sludge (30.6 ± 2.5 mg L(-1)). Monod and Luong models were applied to investigate the influence of the initial substrate concentration, and the results suggested that the maximum substrate removal rate would be reduced with Cu(2+) supplementation. Pre-exposure to Cu(2+) could lead to an 18.2-46.2% decrease in the SDA and decreasing percentage of the SDA increased with both exposure time and concentration. In the continuous-flow test, Cu(2+) concentration varied from 1 to 75 mg L(-1); however, no clear deterioration was observed in the reactor, and the reactor was kept stable, with the total nitrogen removal efficiency and total organic carbon efficiency greater than 89.0 and 85.0%, respectively. The results demonstrated the short-term inhibition of Cu(2+) upon denitrification, and no notable adversity was observed during the continuous-flow test after long-term acclimation. PMID:26610098

  16. Factors influencing spatial patterns of activity of denitrifying bacteria in a grassland soil

    Čuhel, Jiří; Šimek, Miloslav; Saby, N.; Chéneby, D.; Philippot, L.

    Istanbul : Turkish Microbiological Society, 2008. s. 314. [Meetings of the Three Divisions of the International Union of Microbiological Societies 2008. International Congress of Bacteriology and Applied Microbiology /12./. 05.08.2008-09.08.2008, Istanbul] R&D Projects: GA AV ČR IAA600660605; GA MŠk LC06066; GA MŠk MEB020726 Institutional research plan: CEZ:AV0Z60660521 Keywords : spatial patterns * denitrifying bacteria * grassland soil Subject RIV: EH - Ecology, Behaviour

  17. Denitrifying microbial consortium in pasture soil is influenced by the intensity of cattle-induced stress

    Čuhel, Jiří; Šimek, Miloslav

    Nijmegen : Radboud University Nijmegen, 2006. s. 38. [COST Action 856, Ecological Aspects of Denitrification , with Emphasis on Agriculture. Management Committee Meeting & Workshop, New developments in the N-cycle reactions in the first decade of the 21st century. 09.03.2006-11.03.2006, Nijmegen] Institutional research plan: CEZ:AV0Z60660521 Keywords : denitrifying microbial consortium * pasture soil * intensity of cattle-induced stress Subject RIV: EH - Ecology, Behaviour

  18. Mapping field-scale spatial patterns of size and activity of the denitrifier community

    Philippot, L.; Čuhel, Jiří; Saby, N.P.A.; Chéneby, D.; Chroňáková, Alica; Bru, D.; Arrouays, D.; Martin-Laurent, F.; Šimek, Miloslav

    2009-01-01

    Roč. 11, č. 6 (2009), s. 1518-1526. ISSN 1462-2912 R&D Projects: GA MŠk MEB020726; GA MŠk LC06066; GA AV ČR IAA600660605 Institutional research plan: CEZ:AV0Z60660521 Keywords : field-scale spatial patterns * denitrifier community Subject RIV: EH - Ecology, Behaviour Impact factor: 4.909, year: 2009

  19. Evaluation of the microbial diversity of denitrifying bacteria in batch reactor

    S. I. Maintinguer; I. K. Sakamoto; M. A. T. Adorno; M. B. A. Varesche

    2013-01-01

    Microbial communities in an industrial activated sludge plant may contribute to the denitrification process, but the information on the microorganisms present in denitrifying reactors is still scarce. Removal of inorganic nitrogen compounds can be accomplished by the addition of carbon sources to the biological process of denitrification. Ethanol is an economically viable alternative as a carbon source in tropical countries like Brazil, with large-scale production from sugarcane. This paper r...

  20. Coexistence of nitrifying, anammox and denitrifying bacteria in a sequencing batch reactor

    MichelaLangone; JiaYan

    2014-01-01

    Elevated nitrogen removal efficiencies from ammonium-rich wastewaters have been demonstrated by several applications, that combine nitritation and anammox processes. Denitrification will occur simultaneously when organic carbon is also present. In this study, the activity of aerobic ammonia oxidizing, anammox and denitrifying bacteria in a full scale Sequencing Batch Reactor, treating digester supernatants, was studied by means of batch-assays. AOB and anammox activities were maximum at pH of...

  1. Abundance, composition and activity of denitrifier communities in metal polluted paddy soils

    Yuan Liu; Yongzhuo Liu; Huimin Zhou; Lianqing Li; Jinwei Zheng; Xuhui Zhang; Jufeng Zheng; Genxing Pan

    2016-01-01

    Denitrification is one of the most important soil microbial processes leading to the production of nitrous oxide (N2O). The potential changes with metal pollution in soil microbial community for N2O production and reduction are not well addressed. In this study, topsoil samples were collected both from polluted and non-polluted rice paddy fields and denitrifier communities were characterized with molecular fingerprinting procedures. All the retrieved nirK sequences could be grouped into neith...

  2. Localization of the cytochrome cd1 and copper nitrite reductases in denitrifying bacteria.

    Coyne, M S; Arunakumari, A; Pankratz, H S; Tiedje, J. M.

    1990-01-01

    The locations of cytochrome cd1 nitrite reductases in Pseudomonas aeruginosa and Pseudomonas fluorescens and copper nitrite reductases in Achromobacter cycloclastes and Achromobacter xylosoxidans were identified. Immunogold labeling with colloidal-gold probes showed that the nitrite reductases were synthesized exclusively in anaerobically grown (denitrifying) cells. Little immunogold label occurred in the cytoplasm of these four strains; most was found in the periplasmic space or was associat...

  3. Impact of plant identity, diversity and composition on diversity, composition and function of nirK-type denitrifying microorganisms in temperate grassland soil

    Bremer, Christina

    2007-01-01

    The impact of plant species, the number of plant species (diversity) and the composition of plant species on nirK-type denitrifying microorganisms in soil was studied. NirK encodes the enzyme nitrite reductase which is the key enzyme for dissimilatory nitrate reduction. As a measure for denitrifier function, net and gross dinitrogenoxide (N2O) production rates and denitrifier enzyme activity were determined. Plant identity, divers...

  4. Abundance, composition and activity of denitrifier communities in metal polluted paddy soils

    Liu, Yuan; Liu, Yongzhuo; Zhou, Huimin; Li, Lianqing; Zheng, Jinwei; Zhang, Xuhui; Zheng, Jufeng; Pan, Genxing

    2016-01-01

    Denitrification is one of the most important soil microbial processes leading to the production of nitrous oxide (N2O). The potential changes with metal pollution in soil microbial community for N2O production and reduction are not well addressed. In this study, topsoil samples were collected both from polluted and non-polluted rice paddy fields and denitrifier communities were characterized with molecular fingerprinting procedures. All the retrieved nirK sequences could be grouped into neither ?- nor ?- proteobacteria, while most of the nosZ sequences were affiliated with ?-proteobacteria. The abundances of the nirK and nosZ genes were reduced significantly in the two polluted soils. Thus, metal pollution markedly affected composition of both nirK and nosZ denitrifiers. While the total denitrifying activity and N2O production rate were both reduced under heavy metal pollution of the two sites, the N2O reduction rate showed no significant change. These findings suggest that N2O production activity could be sensitive to heavy metal pollution, which could potentially lead to a decrease in N2O emission in polluted paddies. Therefore, metal pollution could have potential impacts on soil N transformation and thus on N2O emission from paddy soils. PMID:26739424

  5. [Analysis on Diversity of Denitrifying Microorganisms in Sequential Batch Bioreactor Landfill].

    Li, Wei-Hua; Sun, Ying-Jie; Liu, Zi-Liang; Ma, Qiang; Yang, Qiang

    2016-01-15

    A denitrification functional microorganism gene clone library (amoA, nosZ) and the PCR-RFLP technology was constructed to investigate the microbial diversity of denitrifying microorganisms in the late period of stabilization of sequential batch bioreactor landfill. The results indicated that: the bacterial diversity of ammonia oxidizing bacteria in the aged refuse reactor was very high, and most of them were unknown groups, also, all bacteria were unculturable or had not been isolated. The phylogenetic analysis suggested that the dominant ammonia oxidizing bacteria were presumably Nitrosomonas of 6-Proteobacteria. The diversity of denitrifying bacteria in fresh refuse reactor was abundant, which mainly included Thauera and Thiobacillus of 6-Proteobacteria. As Thauera sp. has the denitrification characteristics under the condition of aerobic while Thiobacillus denitrificans has the autotrophic denitrification characteristics, it was speculated that aerobic denitrification and autotrophic denitrification might be the main pathways for nitrogen removal in the fresh refuse reactor at the late period of stabilization. Additionally, another group in the gene clone library of denitrifying bacteria may be classified as Bradyrhizobiaceae of alpha-Proteobacteria. PMID:27078976

  6. Abundance, composition and activity of denitrifier communities in metal polluted paddy soils

    Liu, Yuan; Liu, Yongzhuo; Zhou, Huimin; Li, Lianqing; Zheng, Jinwei; Zhang, Xuhui; Zheng, Jufeng; Pan, Genxing

    2016-01-01

    Denitrification is one of the most important soil microbial processes leading to the production of nitrous oxide (N2O). The potential changes with metal pollution in soil microbial community for N2O production and reduction are not well addressed. In this study, topsoil samples were collected both from polluted and non-polluted rice paddy fields and denitrifier communities were characterized with molecular fingerprinting procedures. All the retrieved nirK sequences could be grouped into neither α- nor β- proteobacteria, while most of the nosZ sequences were affiliated with α-proteobacteria. The abundances of the nirK and nosZ genes were reduced significantly in the two polluted soils. Thus, metal pollution markedly affected composition of both nirK and nosZ denitrifiers. While the total denitrifying activity and N2O production rate were both reduced under heavy metal pollution of the two sites, the N2O reduction rate showed no significant change. These findings suggest that N2O production activity could be sensitive to heavy metal pollution, which could potentially lead to a decrease in N2O emission in polluted paddies. Therefore, metal pollution could have potential impacts on soil N transformation and thus on N2O emission from paddy soils.

  7. Characterizations of denitrifying polyphosphate-accumulating bacterium Paracoccus sp. Strain YKP-9.

    Lee, Han Woong; Park, Yong Keun

    2008-12-01

    A denitrifying polyphosphate-accumulating bacterium (YKP-9) was isolated from activated sludge of a 5-stage biological nutrient removal process with step feed system. This organism was a Gram-negative, coccus-shaped, facultative aerobic chemoorganotroph. It had a respiratory type of metabolism with oxygen, nitrate, and nitrite as terminal electron acceptors. The 16S rRNA gene sequence of strain YKP-9 was most similar to the 16S rRNA gene sequence of Paracoccus sp. OL18 (AY312056) (similarity level, 97%). Denitrifying polyphosphate accumulation by strain YKP-9 was examined under anaerobicanoxic and anaerobic-oxic batch conditions. It was able to use external carbon sources for polyhydroxyalkanoates (PHA) synthesis and to release phosphate under anaerobic condition. It accumulated polyphosphate and grew a little on energy provided by external carbon sources under anoxic condition, but did neither accumulate polyphosphate nor grow in the absence of external carbon sources under anoxic condition. Cells with intracellular PHA cannot accumulate polyphosphate in the absence of external carbon sources under anoxic condition. Under oxic condition, it grew but could not accumulate polyphosphate with external carbon sources. Based on the results from this study, strain YKP-9 is a new-type denitrifying polyphosphate-accumulating bacterium that accumulates polyphosphate only under anoxic condition, with nitrate and nitrite as the electron acceptors in the presence of external carbon sources. PMID:19131700

  8. Disentangling the rhizosphere effect on nitrate reducers and denitrifiers: insight into the role of root exudates.

    Henry, S; Texier, S; Hallet, S; Bru, D; Dambreville, C; Chèneby, D; Bizouard, F; Germon, J C; Philippot, L

    2008-11-01

    To determine to which extent root-derived carbon contributes to the effects of plants on nitrate reducers and denitrifiers, four solutions containing different proportions of sugar, organic acids and amino acids mimicking maize root exudates were added daily to soil microcosms at a concentration of 150 microg C g(-1) of soil. Water-amended soils were used as controls. After 1 month, the size and structure of the nitrate reducer and denitrifier communities were analysed using the narG and napA, and the nirK, nirS and nosZ genes as molecular markers respectively. Addition of artificial root exudates (ARE) did not strongly affect the structure or the density of nitrate reducer and denitrifier communities whereas potential nitrate reductase and denitrification activities were stimulated by the addition of root exudates. An effect of ARE composition was also observed on N(2)O production with an N(2)O:(N(2)O + N(2)) ratio of 0.3 in microcosms amended with ARE containing 80% of sugar and of 1 in microcosms amended with ARE containing 40% of sugar. Our study indicated that ARE stimulated nitrate reduction or denitrification activity with increases in the range of those observed with the whole plant. Furthermore, we demonstrated that the composition of the ARE affected the nature of the end-product of denitrification and could thus have a putative impact on greenhouse gas emissions. PMID:18393993

  9. A comparative study of the bacterial community in denitrifying and traditional enhanced biological phosphorus removal processes.

    Lv, Xiao-Mei; Shao, Ming-Fei; Li, Chao-Lin; Li, Ji; Gao, Xin-Lei; Sun, Fei-Yun

    2014-09-17

    Denitrifying phosphorus removal is an attractive wastewater treatment process due to its reduced carbon source demand and sludge minimization potential. Two lab-scale sequencing batch reactors (SBRs) were operated in alternating anaerobic-anoxic (A-A) or anaerobic-oxic (A-O) conditions to achieve denitrifying enhanced biological phosphate removal (EBPR) and traditional EBPR. No significant differences were observed in phosphorus removal efficiencies between A-A SBR and A-O SBR, with phosphorus removal rates being 87.9% and 89.0% respectively. The community structures in denitrifying and traditional EBPR processes were evaluated by high-throughput sequencing of the PCR-amplified partial 16S rRNA genes from each sludge. The results obtained showed that the bacterial community was more diverse in A-O sludge than in A-A sludge. Taxonomy and ?-diversity analyses indicated that a significant shift occurred in the dominant microbial community in A-A sludge compared with the seed sludge during the whole acclimation phase, while a slight fluctuation was observed in the abundance of the major taxonomies in A-O sludge. One Dechloromonas-related OTU outside the 4 known Candidatus "Accumulibacter" clades was detected as the main OTU in A-A sludge at the stationary operation, while Candidatus "Accumulibacter" dominated in A-O sludge. PMID:24964811

  10. Effect of CO2 enrichment and high photosynthetic photon flux densities (PPFD) on rubisco and PEP-case activities of in vitro cultured strawberry plants

    Standard growing conditions in vitro (low light and CO2) are not conducive to autotrophy. An experiment was conducted to improve photosynthesis in vitro in the hope of increasing survival in acclimatization. A factorial experiment was elaborated where CO2 and PPFD were supplied to in vitro cultured strawberry plants in the rooting stage. Activities of carboxylating enzymes were determined after 4 weeks of culture. The activities of non-activated and activated rubisco and PEP-Case were measured after extraction of the enzymes and a reaction with NaH14CO3 followed by scintillation counting spectroscopy. High CO2 concentration significantly increased net assimilation rates (NAR) by 165% over the control for both 1650 and 3000 ppm CO2. High PPFD only increased NAR by 12 and 35% for 150 and 250 ?molm-2s-1 respectively over the control. Plants grown at 3000 ppm CO2 had the highest level of chlorophyll/g FW with 97% more than the control. The activity of PEP-Case was the highest at high light levels and high CO2 with rates of 1.65 for 1650 ppm versus 1.22 mmol CO2 mg-1 chl. h-1 at 250 ?molm-2s-1. There was no difference in PEP activity at low light levels. The rubisco activity was lower at 1650 and 3000 ppm CO2. Increases in NAR correlate more closely to the PEP-Case than to Rubisco activity. Physiological significance of high activity of PEP-Case over rubisco will be discussed

  11. Other enrichment related contracts

    In addition to long-term enrichment contracts, DOE has other types of contracts: (1) short-term, fixed-commitment enrichment contract; (2) emergency sales agreement for enriched uranium; (3) feed material lease agreement; (4) enriched uranium storage agreement; and (5) feed material usage agreement

  12. Denitrifying phosphorus removal from municipal wastewater and dynamics of "Candidatus Accumulibacter" and denitrifying bacteria based on genes of ppk1, narG, nirS and nirK.

    Zeng, Wei; Zhang, Jie; Wang, Anqi; Peng, Yongzhen

    2016-05-01

    Relevance of clade-level population dynamics of "Candidatus Accumulibacter" to performance of denitrifying phosphorus (P) removal from municipal wastewater was investigated. Stable denitrifying P removal in anoxic zone of continuous-flow reactor was achieved, accounting for 90% of total P removal. Clades IIC and IIF affiliated with Accumulibacter lineage were the dominant clades during denitrifying P removal, reaching 90% of ppk1 clone library. NarG gene library indicated Gamma and Beta-proteobacteria played an important role in nitrate reduction. Diversity and abundance of nirS library was much more than nirK, and thus became the main functional gene to execute nitrite reduction. Based on abundance of nirS, nirK and ppk1, the ratio of Accumulibacter capable of denitrifying P removal to total Accumulibacter was 22%. No matter whether Accumulibacter had narG gene or not, high abundance of narG at a level of 10(9)cells/(g dried-sludge) promoted nitrate reduced to nitrite, ensuring performance of denitrifying P removal. PMID:26896717

  13. Effect of pyrene on denitrification activity and abundance and composition of denitrifying community in an agricultural soil.

    Guo, Guang-Xia; Deng, Huan; Qiao, Min; Mu, Yu-Jing; Zhu, Yong-Guan

    2011-07-01

    Toxicity of pyrene on the denitrifiers was studied by spiking an agricultural soil with pyrene to a series of concentrations (0-500 mg kg(-1)) followed by dose-response and dynamic incubation experiments. Results showed a positive correlation between potential denitrification activity and copy numbers of denitrifying functional genes (nirK, nirS and nosZ), and were both negatively correlated with pyrene concentrations. Based on the comparison of EC(50) values, denitrifiers harboring nirK, nirS or nosZ gene were more sensitive than denitrification activity, and denitrifiers harboring nirS gene were more sensitive than that harboring nirK or nosZ genes. Seven days after spiking with EC(50) concentration of pyrene, denitrifiers diversity decreased and community composition changed in comparison with the control. Phylogenetic analyses of three genes showed that the addition of pyrene increased the proportion of Bradyrhizobiaceae, Rhodospirillales, Burkholderiales and Pseudomonadales. Some species belonging to these groups were reported to be able to degrade PAHs. PMID:21497968

  14. Beyond Job Enrichment to Employment Enrichment

    Werther, William B., Jr.

    1975-01-01

    Employment enrichment views the total work environment confronting employees as a system consisting of two overlapping areas: worker-job and worker-organization subsystems. Job enrichment has improved the worker-job subsystem. The focus of this article is on methods of improving the worker-organization relationship. (Author/JB)

  15. Nitrogen Removal by a Fungal Aerobic Denitrifier of Penicillium Strain

    Chaocheng Zhao

    2012-01-01

    Full Text Available A kind of aerobic Penicillium that can remove ammonia, nitrite and nitrate was isolated through an improved bromothymol blue (BTB selective culture medium method in this experiment and then the nitrogen removal by the strain was detailedly investigated. The results showed that this strain was able to make use of many kinds of organic carbon compounds as sole carbon source for the removal of the three types of inorganic nitrogen compounds but the way of removal was different. Ammonia was assimilated for forming cell components such as amino acid and protein, different from which, nitrite and nitrate were eliminated by the aid of dual assimilation and denitrification. When the three types of nitrogen coexist, the removal order was as follows: ammonia>nitrite>nitrate. Type of carbon source, initial nitrogen concentration and carbon nitrogen ratio (C/N all had different effect on final solution pH, dry weight, nitrogen removal rate and removal ability of the strain. It was tested that non-polar organic carbon source containing -CH3 group like sucrose was inclined to be used by the strain. When sucrose was carbon source, the optimum C/N of ammonia, nitrite and nitrate removal were separate 4-6, 8-12 and 12-16. In addition, it was demonstrated with calculation that the removal abilities of the above mentioned three nitrogen of the strain were about 50, 60 and 90 mg g-1 respectively, showing its tremendous capability of nitrogen removal.

  16. 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

    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)

  17. In situ detection of denitrifying bacteria by mRNA-targeted nucleic acid probes and catalyzed reporter deposition

    Kofoed, Michael Vedel; Stief, Peter; Poulsen, Morten; Schramm, Andreas

    In situ detection of denitrifying bacteria by mRNA-targeted nucleic acid probes and catalyzed reporter deposition   Michael V.W. Kofoed, Peter Stief, Morten Poulsen, and Andreas Schramm Department of Biological Sciences, Microbiology, University of Aarhus, Denmark Denitrification, the sequential...... reduction of nitrate to dinitrogen gas, is essential for the removal of fixed nitrogen from natural and engineered ecosystems. However, community structure and activity dynamics of denitrifying bacteria in most systems are poorly understood, partially due to difficulties in identifying and quantifying...... reductase) and nosZ (encoding nitrous oxide reductase), to detect nitrate-reducing and completely denitrifying bacteria, respectively. Enzyme-labelled oligonucleotide probes and digoxygenin-labelled polynucleotide probes were evaluated for in situ hybridization in combination with immunochemical detection...

  18. Diversity of Nitrate-Reducing and Denitrifying Bacteria in a Marine Aquaculture Biofilter and their Response to Sulfide

    Krieger, Bärbel; Schwermer, Carsten U.; Rezakhani, Nastaran; Horn, Marcus A.; Gieseke, Armin; Cytryn, Eddie; Dror, Minz; van Rijn, Jaap; Drake, Harold L.; Schramm, Andreas

    2006-01-01

    DIVERSITY OF NITRATE-REDUCING AND DENITRIFYING BACTERIA IN A MARINE AQUACULTURE BIOFILTER AND THEIR RESPONSE TO SULFIDE B.U. Krieger 1,5, C. Schwermer 2, N. Rezakhani 5, M.A. Horn 1, A. Gieseke 2, E. Cytryn 3, D. Minz 3, J. van Rijn 4, H.L. Drake 1, A. Schramm 5 1 Dept. of Ecological Microbiology...... nitrous oxide reductase, trigger a shift from denitrification to dissimilatory nitrate reduction to ammonium (DNRA), or be used as electron donor for nitrate reduction. The goal of this study was to identify and isolate nitrate-reducing and denitrifying bacteria from the biofilter and to investigate their...... Alphaproteobacteria but also including Beta- and Gammaproteobacteria, Bacteroidetes, Firmicutes, and Actinobacteria. The diversity of the isolates was compared to the cultivation-independent diversity of nitrate-reducing and denitrifying bacteria based on narG and nosZ as functional marker genes. Growth experiments...

  19. Fecal Bacteria, Bacteriophage, and Nutrient Reductions in a Full-Scale Denitrifying Woodchip Bioreactor.

    Rambags, Femke; Tanner, Chris C; Stott, Rebecca; Schipper, Louis A

    2016-05-01

    Denitrifying bioreactors using woodchips or other slow-release carbon sources can be an effective method for removing nitrate (NO) from wastewater and tile drainage. However, the ability of these systems to remove fecal microbes from wastewater has been largely uninvestigated. In this study, reductions in fecal indicator bacteria () and viruses (F-specific RNA bacteriophage [FRNA phage]) were analyzed by monthly sampling along a longitudinal transect within a full-scale denitrifying woodchip bioreactor receiving secondary-treated septic tank effluent. Nitrogen, phosphorus, 5-d carbonaceous biochemical oxygen demand (CBOD), and total suspended solids (TSS) reduction were also assessed. The bioreactor demonstrated consistent and substantial reduction of (2.9 log reduction) and FRNA phage (3.9 log reduction) despite receiving highly fluctuating inflow concentrations [up to 3.5 × 10 MPN (100 mL) and 1.1 × 10 plaque-forming units (100 mL) , respectively]. Most of the removal of fecal microbial contaminants occurred within the first meter of the system (1.4 log reduction for ; 1.8 log reduction for FRNA phage). The system was also efficient at removing NO (>99.9% reduction) and TSS (89% reduction). There was no evidence of consistent removal of ammonium, organic nitrogen, or phosphorus. Leaching of CBOD occurred during initial operation but decreased and stabilized at lower values (14 g O m) after 9 mo. We present strong evidence for reliable microbial contaminant removal in denitrifying bioreactors, demonstrating their broader versatility for wastewater treatment. Research on the removal mechanisms of microbial contaminants in these systems, together with the assessment of longevity of removal, is warranted. PMID:27136150

  20. Relating phylogenetic and functional diversity among denitrifiers and quantifying their capacity to predict community functioning

    JoanaFalcão Salles

    2012-06-01

    Full Text Available Genetic diversity of phylogenetic or functional markers is widely used as a proxy of microbial diversity. However, it remains unclear at what extent functional diversity, gene sequence diversity and community functioning are linked. We analysed, for a range of denitrifying bacteria, the relationships between (i the similarity of functional traits evaluated from metabolic profiles (BIOLOG plates or from N2O accumulation patterns on different C sources, and (ii the similarity of phylogenetic (16S rRNA gene or functional (nir gene markers. We also calculated different proxies for the diversity of denitrifier community based on taxa richness, phylogenetic or functional similarities, and evaluated their performance in inferring the functioning of assembled denitrifying communities. For individual strains, the variation in the 16S rRNA gene sequence explained only 10% of the variation in metabolic patterns and were not related to N2O accumulation. The latter was correlated with the similarity of nitrite reductase residues. When nir genes were analysed separately, the similarity in amino acids coded by the nirS genes accounted for 48% of the variance of the observed pattern of N2O accumulation, whereas nirK amino acid residues were unrelated to N2O accumulation. For bacterial assemblages, phylogenetic diversity and mean community dissimilarity, calculated using 16S rRNA gene sequences, and functional diversity measures associated with BIOLOG, predicted poorly the variation in the functioning of assembled communities (below 15%. In contrast, the proxies of functional diversity based on N2O accumulation patterns performed better and explained from 23 to 42% of the variation in community functioning. Amongst those, community niche was the best metric, indicating the importance of complementarity for carbon resources among taxa in the context of denitrification.

  1. Moving Denitrifying Bioreactors beyond Proof of Concept: Introduction to the Special Section.

    Christianson, Laura E; Schipper, Louis A

    2016-05-01

    Denitrifying bioreactors are organic carbon-filled excavations designed to enhance the natural process of denitrification for the simple, passive treatment of nitrate-nitrogen. Research on and installation of these bioreactors has accelerated within the past 10 years, particularly in watersheds concerned about high nonpoint-source nitrate loads and also for tertiary wastewater treatment. This special section, inspired by the meeting of the Managing Denitrification in Agronomic Systems Community at the 2014 Annual Meeting of the American Society of Agronomy, Crop Science Society of America, and Soil Science Society of America, aims to firmly establish that denitrifying bioreactors for treatment of nitrate in drainage waters, groundwater, and some wastewaters have moved beyond the proof of concept. This collection of 14 papers expands the peer-reviewed literature of denitrifying bioreactors into new locations, applications, and environmental conditions. There is momentum behind the pairing of wood-based bioreactors with other media (biochar, corn cobs) and in novel designs (e.g., use within treatment trains or use of baffles) to broaden applicability into new kinds of waters and pollutants and to improve performance under challenging field conditions such as cool early season agricultural drainage. Concerns about negative bioreactor by-products (nitrous oxide and hydrogen sulfide emissions, start-up nutrient flushing) are ongoing, but this translates into a significant research opportunity to develop more advanced designs and to fine tune management strategies. Future research must think more broadly to address bioreactor impacts on holistic watershed health and greenhouse gas balances and to facilitate collaborations that allow investigation of mechanisms within the bioreactor "black box." PMID:27136139

  2. Performance of Agricultural Residue Media in Laboratory Denitrifying Bioreactors at Low Temperatures.

    Feyereisen, Gary W; Moorman, Thomas B; Christianson, Laura E; Venterea, Rodney T; Coulter, Jeffrey A; Tschirner, Ulrike W

    2016-05-01

    Denitrifying bioreactors can be effective for removing nitrate from agricultural tile drainage; however, questions about cold springtime performance persist. The objective of this study was to improve the nitrate removal rate (NRR) of denitrifying bioreactors at warm and cold temperatures using agriculturally derived media rather than wood chips (WC). Corn ( L.) cobs (CC), corn stover (CS), barley ( L.) straw (BS), WC, and CC followed by a compartment of WC (CC+WC) were tested in laboratory columns for 5 mo at a 12-h hydraulic residence time in separate experiments at 15.5 and 1.5°C. Nitrate-N removal rates ranged from 35 to 1.4 at 15.5°C and from 7.4 to 1.6 g N m d at 1.5°C, respectively; NRRs were ranked CC > CC+WC > BS = CS > WC and CC ≥ CC+WC = CS ≥ BS > WC for 15.5 and 1.5°C, respectively. Although NRRs for CC were increased relative to WC, CC released greater amounts of carbon. Greater abundance of nitrous oxide (NO) reductase gene () was supported by crop residues than WC at 15.5°C, and CS and BS supported greater abundance than WC at 1.5°C. Production of NO relative to nitrate removal (NO) was consistently greater at 1.5°C (7.5% of nitrate removed) than at 15.5°C (1.9%). The NO was lowest in CC (1.1%) and CC-WC (0.9%) and greatest in WC (9.7%). Using a compartment of agricultural residue media in series before wood chips has the potential to improve denitrifying bioreactor nitrate removal rates, but field-scale verification is needed. PMID:27136142

  3. Concurrent activity of anammox and denitrifying bacteria in the Black Sea

    JohnB.Kirkpatrick; ClaraA.Fuchsman; EvgeniyYakushev; JamesT.Staley

    2012-01-01

    After the discovery of ANaerobic AMMonium OXidation (anammox) in the Black Sea in 2003, the role of heterotrophic denitrification as the main marine pathway for fixed N loss was questioned. A 3 part, 15 month time series investigating Black Sea nitrite reductase (nirS) mRNA transcripts at a single location was conducted in order to better understand the activity of anammox and denitrifying bacteria. Here we show that both of these groups were active, as well as being concurrent in the lower s...

  4. In Vivo Emission of Dinitrogen by Earthworms via Denitrifying Bacteria in the Gut

    Horn, Marcus A.; Mertel, Ralph; Gehre, Matthias; Kästner, Matthias; Drake, Harold L.

    2006-01-01

    Earthworms emit the greenhouse gas nitrous oxide (N2O), and ingested denitrifiers in the gut appear to be the main source of this N2O. The primary goal of this study was to determine if earthworms also emit dinitrogen (N2), the end product of complete denitrification. When [15N]nitrate was injected into the gut, the earthworms Aporrectodea caliginosa and Lumbricus terrestris emitted labeled N2 (and also labeled N2O) under in vivo conditions; emission of N2 by these two earthworms was relative...

  5. Sediment denitrifier community composition and nirS gene expression investigated with functional gene microarrays

    Francis, C.A.; Jackson, G.A.; Ward, B.B.; Bülow, Steffen

    2008-01-01

    Chesapeake Bay clones, as well as sequences from many other environments. Greatest nirS diversity was detected at the freshwater station at the head of the bay and least diversity at the higher salinity station near the mouth of the Bay. The most common OTUs from the sequence database were detected on the...... total RNA extracts) targets were hybridized to the same array to compare the profiles of community composition at the DNA (relative abundance) and mRNA (gene expression) levels. Only the three dominant denitrifying groups (in terms of relative strength of DNA hybridization signal) were detected at the m...

  6. Adaptation of Denitrifying Populations to Low Soil pH †

    Timothy B. Parkin; Sexstone, Alan J.; Tiedje, James M.

    1985-01-01

    Natural denitrification rates and activities of denitrifying enzymes were measured in an agricultural soil which had a 20-year past history of low pH (pH ca. 4) due to fertilization with acid-generating ammonium salts. The soil adjacent to this site had been limed and had a pH of ca. 6.0. Natural denitrification rates of these areas were of similar magnitude: 158 ng of N g−1 of soil day−1 for the acid soil and 390 ng of N g−1 of soil day−1 at the neutral site. Estimates of in situ denitrifyin...

  7. Derived enriched uranium market

    The potential impact on the uranium market of highly enriched uranium from nuclear weapons dismantling in the Russian Federation and the USA is analyzed. Uranium supply, conversion, and enrichment factors are outlined for each country; inventories are also listed. The enrichment component and conversion components are expected to cause little disruption to uranium markets. The uranium component of Russian derived enriched uranium hexafluoride is unresolved; US legislation places constraints on its introduction into the US market

  8. Enrichment of recycled uranium

    The centrifuge enrichment process is well suited for the re-enrichment of uranium recovered during reprocessing. BNFL has participated fully with Urenco shareholders in studies which have enabled Urenco to offer commercial enrichment services on recycled uranium. The paper examines aspects of chemical and radiological purity of enrichment plant feed and product. Additionally the effects of programming the flow of reprocessed uranium throughout the fuel cycle are examined and the potential economic incentives quantified. (author)

  9. Uranium Enrichment, an overview

    This general presentation on uranium enrichment will be followed by lectures on more specific topics including descriptions of enrichment processes and assessments of the prevailing commercial and industrial situations. I shall therefore avoid as much as possible duplications with these other lectures, and rather dwell on: some theoretical aspects of enrichment in general, underlying the differences between statistical and selective processes, a review and comparison between enrichment processes, remarks of general order regarding applications, the proliferation potential of enrichment. It is noteworthy that enrichment: may occur twice in the LWR fuel cycle: first by enriching natural uranium, second by reenriching uranium recovered from reprocessing, must meet LWR requirements, and in particular higher assays required by high burn up fuel elements, bears on the structure of the entire front part of the fuel cycle, namely in the conversion/reconversion steps only involving UF6 for the moment. (author). tabs., figs., 4 refs

  10. Molecular approaches to understand the regulation of N2O emission from denitrifying bacteria - model strains and soil communities (Invited)

    Frostegard, A.; Bakken, L. R.

    2010-12-01

    Emissions of N2O from agricultural soils are largely caused by denitrifying bacteria. Field measurements of N2O fluxes show large variations and depend on several environmental factors, and possibly also on the composition of the denitrifying microbial community. The temporal and spatial variation of fluxes are not adequately captured by biogeochemical models, and few options for mitigations have been invented, which underscores the need to understand the mechanisms underlying the emissions of N2O. Analyses of denitrification genes and transcripts extracted from soils are important for describing the system, but may have limited value for prediction of N2O emissions. In contrast, phenotypic analyses are direct measures of the organisms responses to changing environmental conditions. Our approach is to combine phenotypic characterizations using high-resolution gas kinetics, with gene transcription analyses to study denitrification regulatory phenotypes (DRP) of bacterial strains or complex microbial communities. The rich data sets obtained provide a basis for refinement of biochemical and physiological research on this key process in the nitrogen cycle. The strength of this combined approach is illustrated by a series of experiments investigating effects of soil pH on denitrification. Soil pH emerges as a master variable determining the microbial community composition as well as its denitrification product ratio (N2O/N2), with higher ratio in acid than in alkaline soil. It is therefore likely that emissions of N2O from agro-ecosystems will increase in large parts of the world where soil pH is decreasing due to intensified management and increased use of chemical fertilizers. Considering its immense implications, surprisingly few attempts have been made to unravel the mechanisms involved in the pH-control of the product stoichiometry of denitrification. We investigated the kinetics of gas transformations (O2, NO, N2O and N2) and transcription of functional genes in intact soil samples from long-term liming experiments. Expression of nirS (encoding nitrite reductase) and nosZ (encoding N2OR) was generally lower at pH6 compared to pH8, but the nosZ/nirS transcript ratios were similar or even higher at pH6. These results were largely corroborated in refined experiments using extracted bacterial communities from the same soils, which allowed us to better control the pH levels experienced by the cells. The findings indicated that the higher N2O/N2 product ratios at pH6 were due to a post-transcriptional effect. Pure culture experiments using Paracoccus denitrificans demonstrated very low N2O/N2 ratios at pH7 while N2O reduction was severely inhibited by suboptimal pH, and that this inhibition occurred during protein synthesis/assembly rather than at the transcription level. The results are important for the understanding of main factors affecting N2O emissions from terrestrial ecosystems.

  11. Quantity-activity relationship of denitrifying bacteria and environmental scaling in streams of a forested watershed

    O'Connor, B.L.; Hondzo, Miki; Dobraca, D.; LaPara, T.M.; Finlay, J.A.; Brezonik, P.L.

    2006-01-01

    The spatial variability of subreach denitrification rates in streams was evaluated with respect to controlling environmental conditions, molecular examination of denitrifying bacteria, and dimensional analysis. Denitrification activities ranged from 0 and 800 ng-N gsed-1 d-1 with large variations observed within short distances (Denitrifying bacteria were quantified using a competitive polymerase chain reaction (cPCR) technique that amplified the nirK gene that encodes for nitrite reductase. Results showed a range of nirK quantities from 103 to 107 gene-copy-number gsed.-1 A nonparametric statistical test showed no significant difference in nirK quantifies among stream reaches, but revealed that samples with a high denitrification potential had significantly higher nirK quantities. Denitrification activity was positively correlated with nirK quantities with scatter in the data that can be attributed to varying environmental conditions along stream reaches. Dimensional analysis was used to evaluate denitrification activities according to environmental variables that describe fluid-flow properties, nitrate and organic material quantities, and dissolved oxygen flux. Buckingham's pi theorem was used to generate dimensionless groupings and field data were used to determine scaling parameters. The resulting expressions between dimensionless NO3- flux and dimensionless groupings of environmental variables showed consistent scaling, which indicates that the subreach variability in denitrification rates can be predicted by the controlling physical, chemical, and microbiological conditions. Copyright 2006 by the American Geophysical Union.

  12. Evaluation of the microbial diversity of denitrifying bacteria in batch reactor

    S. I. Maintinguer

    2013-09-01

    Full Text Available Microbial communities in an industrial activated sludge plant may contribute to the denitrification process, but the information on the microorganisms present in denitrifying reactors is still scarce. Removal of inorganic nitrogen compounds can be accomplished by the addition of carbon sources to the biological process of denitrification. Ethanol is an economically viable alternative as a carbon source in tropical countries like Brazil, with large-scale production from sugarcane. This paper reports the successful aplication of activated sludge with nitrate and ethanol in a batch anaerobic reactor. The operation lasted 61.5 h with total consumption of nitrate in 42.5 h, nitrite generation (2.0 mg/L and ethanol consumption (830.0 mg/L in 23.5 h. Denitrifying cell counts by the most probable number at the start of the operation were lower than at the end, confirming the ability of the inoculum from activated sludge for the denitrification process. The samples from cell counts were identified as Acidovorax sp., Acinetobacter sp., Comamonas sp. and uncultured bacteria. Therefore, these species may be involved in nitrate reduction and ethanol consumption in the batch reactor.

  13. Autotrophic, hydrogen-oxidizing, denitrifying bacteria in groundwater, potential agents for bioremediation of nitrate contamination

    Smith, R.L.; Ceazan, M.L.; Brooks, M.H.

    1994-01-01

    Addition of hydrogen or formate significantly enhanced the rate of consumption of nitrate in slurried core samples obtained from an active zone of denitrification in a nitrate-contaminated sand and gravel aquifer (Cape Cod, Mass.). Hydrogen uptake by the core material was immediate and rapid, with an apparent K(m) of 0.45 to 0.60 ??M and a V(max) of 18.7 nmol cm-3 h-1 at 30??C. Nine strains of hydrogen-oxidizing denitrifying bacteria were subsequently isolated from the aquifer. Eight of the strains grew autotrophically on hydrogen with either oxygen or nitrate as the electron acceptor. One strain grew mixotrophically. All of the isolates were capable of heterotrophic growth, but none were similar to Paracoccus denitrificans, a well-characterized hydrogen-oxidizing denitrifier. The kinetics for hydrogen uptake during denitrification were determined for each isolate with substrate depletion progress curves; the K(m)s ranged from 0.30 to 3.32 ??M, with V(max)s of 1.85 to 13.29 fmol cell-1 h-1. Because these organisms appear to be common constituents of the in situ population of the aquifer, produce innocuous end products, and could be manipulated to sequentially consume oxygen and then nitrate when both were present, these results suggest that these organisms may have significant potential for in situ bioremediation of nitrate contamination in groundwater.

  14. Identification of active denitrifiers in rice paddy soil by DNA- and RNA-based analyses.

    Yoshida, Megumi; Ishii, Satoshi; Fujii, Daichi; Otsuka, Shigeto; Senoo, Keishi

    2012-01-01

    Denitrification occurs markedly in rice paddy fields; however, few microbes that are actively involved in denitrification in these environments have been identified. In this study, we used a laboratory soil microcosm system in which denitrification activity was enhanced. DNA and RNA were extracted from soil at six time points after enhancing denitrification activity, and quantitative PCR and clone library analyses were performed targeting the 16S rRNA gene and denitrification functional genes (nirS, nirK and nosZ) to clarify which microbes are actively involved in denitrification in rice paddy soil. Based on the quantitative PCR results, transcription levels of the functional genes agreed with the denitrification activity, although gene abundance did not change at the DNA level. Diverse denitrifiers were detected in clone library analysis, but comparative analysis suggested that only some of the putative denitrifiers, especially those belonging to the orders Neisseriales, Rhodocyclales and Burkholderiales, were actively involved in denitrification in rice paddy soil. PMID:22972387

  15. Optimizing and real-time control of biofilm formation, growth and renewal in denitrifying biofilter.

    Liu, Xiuhong; Wang, Hongchen; Long, Feng; Qi, Lu; Fan, Haitao

    2016-06-01

    A pilot-scale denitrifying biofilter (DNBF) with a treatment capacity of 600m(3)/d was used to study real-time control of biofilm formation, removal and renewal. The results showed biofilm formation, growth and removal can be well controlled using on-line monitored turbidity. The status of filter layer condition can be well indicated by Turb break points on turbidity profile. There was a very good linear relationship between biofilm growth degree (Xbiof) and filter clogging degree (Cfilter) with R(2) higher than 0.99. Filter layer clogging coefficient (Yc) lower than 0.27 can be used to determine stable filter layer condition. Since variations of turbidity during backwash well fitted normal distribution with R(2) higher than 0.96, biofilm removal during backwash also can be well optimized by turbidity. Although biofilm structure and nirK-coding denitrifying communities using different carbon sources were much more different, DNBF was still successfully and stably optimized and real-time controlled via on-line turbidity. PMID:26994461

  16. Enrichment of high ammonia tolerant methanogenic culture

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

    2012-01-01

    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, co...

  17. Importance of pre-enrichment media for isolation of Salmonella spp. from swine and poultry

    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......, it may be advantageous to use SEE rather than standard culturing. (C) 1998 Federation of European Microbiological Societies....

  18. EFFECTS OF A PERMEABLE REACTIVE BARRIER ON DENITRIFYING BACTERIA COMMUNITIES AND METHYLMERCURY CONCENTRATIONS IN WAQUOIT BAY, MA

    It is expected that microbial denitrifying bacterial communities in the barrier will be distinct from those at control sites due to the high amount of degradable carbon and the unique redox conditions present within the barrier. It also is expected that toxic methylmercury ...

  19. Diversity of nitrite reductase genes (nirS) in the denitrifying water column of the coastal Arabian Sea

    Jayakumar, D.A.; Francis, C.A.; Naqvi, S.W.A.; Ward, B.B.

    and nitrate distribution in the Arabian Sea coastal denitrifying region are reported. Phylogenetic analysis grouped 132 nirS Arabian Sea sequences into 12 major clusters. Most of the nirS sequences from the coastal water column did not show a high level...

  20. Advanced enrichment techniques

    BNFL is in a unique position in that it has commercial experience of diffusion enrichment, and of centrifuge enrichment through its associate company Urenco. In addition BNFL is developing laser enrichment techniques as part of a UK development programme in this area. The paper describes the development programme which led to the introduction of competitive centrifuge enrichment technology by Urenco and discusses the areas where improvements have and will continue to be made in the centrifuge process. It also describes the laser development programme currently being undertaken in the UK. The paper concludes by discussing the relative merits of the various methods of uranium enrichment, with particular reference to the enrichment market likely to obtain over the rest of the century. (author)

  1. Alternative isotope enrichment processes

    Alternative processes such as gas centrifugation, plasma separation, and laser excited separation are evaluated for use at the ORNL Stable Isotope Enrichment Facility. The applicabiliy of each process to the isotopic enrichment of the calutron feed material and to the selective production of isotopes is determined. The process energy demands are compared to those of the existing facilities. The isotopic enrichment of the feed material prior to a first-pass through the calutrons can result in a significant saving in energy

  2. Uranium enrichment: an overview

    This paper is a general presentation of uranium enrichment processes and assessments of the prevailing commercial and industrial situations. It gives first some theoretical aspects of enrichment in general and explains the differences between statistical and selective processes in particular. Then a review of the different processes is made with a comparison between them. Finally, some general remarks concerning applications are given and the risks of proliferation related to enrichment are mentioned. (J.S.). 4 refs., 5 figs., 8 tabs

  3. Uranium enrichment in Japan

    A uranium enrichment pilot plant that uses the gas centrifugation process came into full operation in 1982. Japanese utilities expect the demand for enrichment services to be about 8 million SWU/y for 1990 and 12 million SWU/y for 2000. Although most enrichment services needed up to the year 2000 have been secured, domestic production capability is needed to ensure security of supply. Japan will proceed to commission a commercial enrichment plant by 1990; its capacity should reach 3 million SWU/y by 2000. Research continues into improvements in the centrifuge process

  4. A Cultural Classroom Library

    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…

  5. Denitrifying sulfide removal process on high-salinity wastewaters in the presence of Halomonas sp.

    Liu, Chunshuang; Zhao, Dongfeng; Ma, Wenjuan; Guo, Yadong; Wang, Aijie; Wang, Qilin; Lee, Duu-Jong

    2016-02-01

    Biological conversion of sulfide, acetate, and nitrate to, respectively, elemental sulfur (S(0)), carbon dioxide, and nitrogen-containing gas (such as N2) at NaCl concentration of 35-70g/L was achieved in an expanded granular sludge bed (EGSB) reactor. A C/N ratio of 1:1 was noted to achieve high sulfide removal and S(0) conversion rate at high salinity. The extracellular polymeric substance (EPS) quantities were increased with NaCl concentration, being 11.4-mg/g volatile-suspended solids at 70mg/L NaCl. The denitrifying sulfide removal (DSR) consortium incorporated Thauera sp. and Halomonas sp. as the heterotrophs and Azoarcus sp. being the autotrophs at high salinity condition. Halomonas sp. correlates with the enhanced DSR performance at high salinity. PMID:26454867

  6. Commercial developments in enrichment

    The significance of the dollars spent by government and industry so far on enrichment programs is enhanced by the number of engineering and component design and supply firms who have become involved in the public and private development and design projects. The knowhow gained from this experience provides the nucleus of the industrial manufacturing base for the unique enrichment technology necessary to build the future U.S. enrichment plants. The $1.5 billion CIP/CUP programs now under way and the $4.4 billion Portsmouth Add-On Plant are bringing the U.S. enrichment capacity to the capability to supply the contracted 323 GW of reactors, while conserving uranium by allowing operation at a lower, more nearly optimum tails assay. The government enrichment contract books have remained closed to new orders since July of 1975. Inevitably, new foreign enrichment projects are actively pursuing customers. Until the government or private U.S. enrichment ventures resume contracting activity, the foreign plants have no U.S. competition. The technology exists within ERDA and industry to maintain U.S. leadership in the world enrichment market if the institutional questions can be answered

  7. Modeling of Cr(VI) Bioreduction Under Fermentative and Denitrifying Conditions

    Molins, S.; Steefel, C.; Yang, L.; Beller, H. R.

    2011-12-01

    The mechanisms of bioreductive immobilization of Cr(VI) were investigated by reactive transport modeling of a set of flow-through column experiments performed using natural Hanford 100H aquifer sediment. The columns were continuously eluted with 5 μM Cr(VI), 5 mM lactate as the electron donor, and selected electron acceptors (tested individually). Here we focus on the two separate experimental conditions that showed the most removal of Cr(VI) from solution: fermentation and denitrification. In each case, a network of enzymatic and abiotic reaction pathways was considered to interpret the rate of chromate reduction. The model included biomass growth and decay, and thermodynamic limitations on reaction rates, and was constrained by effluent concentrations measured by IC and ICP-MS and additional information from bacterial isolates from column effluent. Under denitrifying conditions, Cr(VI) reduction was modeled as co-metabolic with nitrate reduction based on experimental observations and previous studies on a denitrifying bacterium derived from the Hanford 100H aquifer. The reactive transport model results supported this interpretation of the reaction mechanism and were used to quantify the efficiency of the process. The models results also suggest that biomass growth likely relied on a nitrogen source other than ammonium (e.g. nitrate). Under fermentative conditions and based on cell suspension studies performed on a bacterial isolate from the columns, the model assumes that Cr(VI) reduction is carried out directly by fermentative bacteria that convert lactate into acetate and propionate. The evolution to complete lactate fermentation and Cr(VI) reduction took place over a week's time and simulations were used to determine an estimate for a lower limit of the rate of chromate reduction by calibration with the flow-through column experimental results. In spite of sulfate being added to these columns, sulfate reduction proceeded at a slow rate and was not well constrained.

  8. [Denitrifying bacteria of constructed wetland system based on nitrous oxide reductase gene (nosZ)].

    Wang, Xiao-Jun; Chen, Shao-Hua; Zhang, Zhao-Ji; Xiao, Jun-Chao

    2012-04-01

    A horizontal subsurface flow constructed wetland was used to treat micro-pollution surface water and its effluent flowed into a landscape river as the water supplement. The abundance of the nitrous oxide reductase gene nosZ and 16S rDNA was investigated for the rhizosphere soil of the constructed wetland and the sediment of the landscape river by quantitive PCR. The diversity and similarity of denitrifiers were studied by using PCR-DGGE technique targeting the functional gene nosZ. The quantitive PCR results showed that the copy number of 16S rDNA and nosZ gene of the rhizosphere soil of the constructed wetland and the sediment of the landscape river were 1.91E + 07, 1.26E + 06 and 2.68E + 07, 8.37E + 05 copies x ng(-1) (by DNA), and 1.45E + 11, 9.31E + 09 and 5.31E + 11, 1.45E + 10 copies x g(-1) (by dry soil), respectively. Generally, the total bacterial count and denitrifiers in the sediment of the landscape river was higher than that of the constructed wetland, but the relative abundance of nosZ of the latter (3.8% - 10.1%) was significantly higher than that of the former (1.7% - 4.1%). There was low similarity between the two niches in cluster analysis. The predominant species of rhizosphere soil were mainly contained Rhodobacearales, Rhizobiales and Burkholderiales, although the predominant species of sediment were uncultured bacteria. PMID:22720582

  9. Coexistence of nitrifying, anammox and denitrifying bacteria in a sequencing batch reactor.

    Langone, Michela; Yan, Jia; Haaijer, Suzanne C M; Op den Camp, Huub J M; Jetten, Mike S M; Andreottola, Gianni

    2014-01-01

    Elevated nitrogen removal efficiencies from ammonium-rich wastewaters have been demonstrated by several applications, that combine nitritation and anammox processes. Denitrification will occur simultaneously when organic carbon is also present. In this study, the activity of aerobic ammonia oxidizing, anammox and denitrifying bacteria in a full scale sequencing batch reactor, treating digester supernatants, was studied by means of batch-assays. AOB and anammox activities were maximum at pH of 8.0 and 7.8-8.0, respectively. Short term effect of nitrite on anammox activity was studied, showing nitrite up to 42 mg/L did not result in inhibition. Both denitrification via nitrate and nitrite were measured. To reduce nitrite-oxidizing activity, high NH3-N (1.9-10 mg NH3-N/L) and low nitrite (3-8 mg TNN/L) are required conditions during the whole SBR cycle. Molecular analysis showed the nitritation-anammox sludge harbored a high microbial diversity, where each microorganism has a specific role. Using ammonia monooxygenase ?-subunit (amoA) gene as a marker, our analyses suggested different macro- and micro-environments in the reactor strongly affect the AOB community, allowing the development of different AOB species, such as N. europaea/eutropha and N. oligotropha groups, which improve the stability of nitritation process. A specific PCR primer set, used to target the 16S rRNA gene of anammox bacteria, confirmed the presence of the "Ca. Brocadia fulgida" type, able to grow in presence of organic matter and to tolerate high nitrite concentrations. The diversity of denitrifiers was assessed by using dissimilatory nitrite reductase (nirS) gene-based analyses, who showed denitifiers were related to different betaproteobacterial genera, such as Thauera, Pseudomonas, Dechloromonas and Aromatoleum, able to assist in forming microbial aggregates. Concerning possible secondary processes, no n-damo bacteria were found while NOB from the genus Nitrobacter was detected. PMID:24550899

  10. Continuous nitrous oxide abatement in a novel denitrifying off-gas bioscrubber.

    Frutos, Osvaldo D; Arvelo, Ilan A; Pérez, Rebeca; Quijano, Guillermo; Muñoz, Raúl

    2015-04-01

    The potential of a bioscrubber composed of a packed bed absorption column coupled to a stirred tank denitrification bioreactor (STR) was assessed for 95 days for the continuous abatement of a diluted air emission of N2O at different liquid recycling velocities. N2O removal efficiencies of up to 40 ± 1 % were achieved at the highest recirculation velocity (8 m h(-1)) at an empty bed residence time of 3 min using a synthetic air emission containing N2O at 104 ± 12 ppmv. N2O was absorbed in the packed bed column and further reduced in the STR at efficiencies >80 % using methanol as electron donor. The long-term operation of the bioscrubber suggested that the specialized N2O degrading community established was not able to use N2O as nitrogen source. Additional nitrification assays showed that the activated sludge used as inoculum was not capable of aerobically oxidizing N2O to nitrate or nitrite, regardless of the inorganic carbon concentration tested. Denitrification assays confirmed the ability of non-acclimated activated sludge to readily denitrify N2O at a specific rate of 3.9 mg N2O g VSS h(-1) using methanol as electron donor. This study constitutes, to the best of our knowledge, the first systematic assessment of the continuous abatement of N2O in air emission. A characterization of the structure of the microbial population in the absorption column by DGGE-sequencing revealed a high microbial diversity and the presence of heterotrophic denitrifying methylotrophs. PMID:25547842

  11. Uranium enrichment. Principles

    Uranium enrichment industry is a more than 60 years old history and has developed without practically no cost, efficiency or profit constraints. However, remarkable improvements have been accomplished since the Second World War and have led to the development of various competing processes which reflect the diversity of uranium compositions and of uranium needs. Content: 1 - general considerations: uranium isotopes, problem of uranium enrichment, first realizations (USA, Russia, Europe, Asia, other countries), present day situation, future needs and market evolution; 2 - principles of isotopic separation: processes classification (high or low enrichment), low elementary enrichment processes, equilibrium time, cascade star-up and monitoring, multi-isotopes case, uranium reprocessing; 3 - enrichment and proliferation. (J.S.)

  12. Input Deficit and Stimulus Enrichment: A Replication-with-Expansion

    Friedrich, Douglas; And Others

    1973-01-01

    Two studies were conducted based on a methodological criticism of J. Gordon and H. Haywood's (1969) research on stimulus enrichment procedures with cultural-familial and brain-damaged retarded persons. (Author)

  13. Developments in uranium enrichment

    The enrichment services market is still characterized by overcapacities. While consumption worldwide will rise by some 15% to 39,000 t SWU/a over the next ten years, capacities amount to nearly 50,000 t SWU/a. The price for enrichment services probably has reached its all time low. Prices below U.S. $ 100/kg SWU are not likely to cover costs even of the economically most advanced enrichment processes. Urenco has prepared for the difficult enrichment business in the years to come by streamlining and cost cutting measures. The company intends to hold and increase its share of more than 10% in the world market. The uranium enrichment plant of Gronau will be expanded further. Expansion beyond 1000 t is subject to another permit being granted under the Atomic Energy Act, an application for which was filed in December 1994. Centrifuge technology is the superior enrichment technology, i.e., there is still considerable potential for further development. Construction of enrichment plants employing the centrifuge technology in the United States and in France is being pursued in various phases, from feasibility studies to licensing procedures. Before these plants could be implemented, however, considerable problems of organization would have to be solved, and the market would have to change greatly, respectively. The laser process, at the present time, does not seem to be able to develop into a major industrial competitor. (orig.)

  14. TRIGA low enrichment fuel

    Sixty TRIGA reactors have been sold and the earliest of these are now passing twenty years of operation. All of these reactors use the uranium-zirconium hydride fuel (UZrH) which provides certain unique advantages arising out of its large prompt negative temperature coefficient, very low fission product release, and high temperature capability. Eleven of these Sixty reactors are conversions from plate fuel to TRIGA fuel which were made as a result of these advantages. With only a few exceptions, TRIGA reactors have always used low-enriched-uranium (LEU) fuel with an enrichment of 19.9%. The exceptions have either been converted from the standard low-enriched fuel to the 70% enriched FLIP fuel in order to achieve extended lifetime, or are higher powered reactors which were designed for long life using 93%-enriched uranium during the time when the use and export of highly enriched uranium (HEU) was not restricted. The advent of international policies focusing attention on nonproliferation and safeguards made the HEU fuels obsolete. General Atomic immediately undertook a development effort (nearly two years ago) in order to be in a position to comply with these policies for all future export sales and also to provide a low-enriched alternative to fully enriched plate-type fuels. This important work was subsequently partially supported by the U.S. Department of Energy. The laboratory and production tests have shown that higher uranium densities can be achieved to compensate for reducing the enrichment to 20%, and that the fuels maintain the characteristics of the very thoroughly proven standard TRIGA fuels. In May of 1978, General Atomic announced that these fuels were available for TRIGA reactors and for plate-type reactors with power levels up to 15 MW with GA's standard commercial warranty

  15. TRIGA low enrichment fuel

    Sixty TRIGA reactors have been sold and the earliest of these are now passing twenty years of operation. All of these reactors use the uranium zirconium hydride fuel (UZrH) which provides certain unique advantages arising out of its large prompt negative temperature coefficient, very low fission product release, and high temperature capability. Eleven of these Sixty reactors are conversions from plate fuel to TRIGA fuel which were made as a result of these advantages. With only a few exceptions, TRIGA reactors have always used low-enriched uranium (LEU) fuel with an enrichment of 19.9%. The exceptions have either been converted from the standard low-enriched fuel to the 70% enriched FLIP fuel in order to achieve extended lifetime, or are higher powered reactors which were designed for long life using 93%-enriched uranium during the time when the use and export of highly enriched uranium (HEU) was not restricted. The advent of international policies focusing attention on nonproliferation and safeguards made the HEU fuels obsolete. General Atomic immediately undertook a development effort (nearly two years ago) in order to be in a position to comply with these policies for all future export sales and also to provide a low-enriched alternative to fully enriched plate-type fuels. This important work was subsequently partially supported by the U.S. Department of Energy. The laboratory and production tests have shown that higher uranium densities can be achieved to compensate for reducing the enrichment to 20%, and that the fuels maintain the characteristics of the very thoroughly proven standard TRIGA fuels. In May of 1978, General Atomic announced that these fuels were available for TRIGA reactors and for plate-type reactors with power levels up to 15 MW with General Atomic's standard commercial warranty

  16. Uranium conversion and enrichment

    A description is given of the Atomic Energy Corporation's uranium conversion and enrichment plants at Valinda ba, including a brief discussion of problems encountered and plans for future developments. (author)

  17. Metal enrichment processes

    Schindler, S

    2008-01-01

    There are many processes that can transport gas from the galaxies to their environment and enrich the environment in this way with metals. These metal enrichment processes have a large influence on the evolution of both the galaxies and their environment. Various processes can contribute to the gas transfer: ram-pressure stripping, galactic winds, AGN outflows, galaxy-galaxy interactions and others. We review their observational evidence, corresponding simulations, their efficiencies, and their time scales as far as they are known to date. It seems that all processes can contribute to the enrichment. There is not a single process that always dominates the enrichment, because the efficiencies of the processes vary strongly with galaxy and environmental properties.

  18. [Effect of short-time drought process on denitrifying bacteria abundance and N2O emission in paddy soil].

    Lu, Jing; Liu, Jin-Bo; Sheng, Rong; Liu, Yi; Chen, An-Lei; Wei, Wen-Xue

    2014-10-01

    In order to investigate the impact of drying process on greenhouse gas emissions and denitrifying microorganisms in paddy soil, wetting-drying process was simulated in laboratory conditions. N2O flux, redox potential (Eh) were monitored and narG- and nosZ-containing denitrifiers abundances were determined by real-time PCR. N2O emission was significantly increased only 4 h after drying process began, and it was more than 6 times of continuous flooding (CF) at 24 h. In addition, narG and nosZ gene abundances were increased rapidly with the drying process, and N2O emission flux was significantly correlated with narG gene abundance (P paddy soil. PMID:25796895

  19. Motif Enrichment Tool

    Blatti, Charles; Sinha, Saurabh

    2014-01-01

    The Motif Enrichment Tool (MET) provides an online interface that enables users to find major transcriptional regulators of their gene sets of interest. MET searches the appropriate regulatory region around each gene and identifies which transcription factor DNA-binding specificities (motifs) are statistically overrepresented. Motif enrichment analysis is currently available for many metazoan species including human, mouse, fruit fly, planaria and flowering plants. MET also leverages high-thr...

  20. Metal enrichment processes

    Schindler, S.; Diaferio, A.

    2008-01-01

    There are many processes that can transport gas from the galaxies to their environment and enrich the environment in this way with metals. These metal enrichment processes have a large influence on the evolution of both the galaxies and their environment. Various processes can contribute to the gas transfer: ram-pressure stripping, galactic winds, AGN outflows, galaxy-galaxy interactions and others. We review their observational evidence, corresponding simulations, their efficiencies, and the...

  1. Structure and function of microbial communities in constructed wetlands - influence of environmental parameters and pesticides on denitrifying bacteria

    Milenkovski, Susann

    2009-01-01

    This thesis addresses the interactions and relationships between natural aquatic bacterial communities, environmental parameters, anthropogenic chemicals and the denitrification pathway in the habitat of agricultural constructed wetlands. The main aim was to gain fundamental knowledge of the drivers behind the processes of the denitrification (i.e. nitrogen removal) in constructed wetlands, hence, the structure and function of the denitrifying bacterial community as efficient nitrogen removal...

  2. Is soil fauna an important driver of key soil function ? the case of denitrifier communities in tropical soils

    Brauman, Alain; Philippot, Laurent; Baudoin, Ezékiel; Djigal, Djibril; Gueye, M.; Rabary, B.; Assigbetse, K.; Lensi, R.; Villenave, Cécile; Blanchart, Eric

    2008-01-01

    According to recent estimations, soil animals represent as much as 23% of the total diversity of living organisms. If the importance of soil fauna on soil global microbial processes like mineralization is widely recognized, their impact on composition of soil microbial guilds, like denitrifiers, remains questionable. This study will resume several works undertaken on the influence of keystone tropical soil species (nematodes, termites, earthworms) on different parameters (activity- density an...

  3. Pyruvic Oxime Nitrification and Copper and Nickel Resistance by a Cupriavidus pauculus, an Active Heterotrophic Nitrifier-Denitrifier

    Miguel Ramirez; Jennifer Obrzydowski; Mary Ayers; Sonia Virparia; Meijing Wang; Kurtis Stefan; Richard Linchangco; Domenic Castignetti

    2014-01-01

    Heterotrophic nitrifiers synthesize nitrogenous gasses when nitrifying ammonium ion. A Cupriavidus pauculus, previously thought an Alcaligenes sp. and noted as an active heterotrophic nitrifier-denitrifier, was examined for its ability to produce nitrogen gas (N2) and nitrous oxide (N2O) while heterotrophically nitrifying the organic substrate pyruvic oxime [CH3C(NOH)COOH]. Neither N2 nor N2O were produced. Nucleotide and phylogenetic analyses indicated that the organism is a member of a ge...

  4. Anaerobic Mineralization of Quaternary Carbon Atoms: Isolation of Denitrifying Bacteria on Pivalic Acid (2,2-Dimethylpropionic Acid)

    Probian, Christina; Wülfing, Annika; Harder, Jens

    2003-01-01

    The degradability of pivalic acid was established by the isolation of several facultative denitrifying strains belonging to Zoogloea resiniphila, to Thauera and Herbaspirillum, and to Comamonadaceae, related to [Aquaspirillum] and Acidovorax, and of a nitrate-reducing bacterium affiliated with Moraxella osloensis. Pivalic acid was completely mineralized to carbon dioxide. The catabolic pathways may involve an oxidation to dimethylmalonate or a carbon skeleton rearrangement, a putative 2,2-dim...

  5. Population analysis in a denitrifying sand filter: conventional and in situ identification of Paracoccus spp. in methanol-fed biofilms.

    Neef, A.; Zaglauer, A; Meier, H.; Amann, R; Lemmer, H; Schleifer, K H

    1996-01-01

    The microbial community of a denitrifying sand filter in a municipal wastewater treatment plant was examined by conventional and molecular techniques to identify the bacteria actively involved in the removal of nitrate. In this system, denitrification is carried out as the last step of water treatment by biofilms growing on quartz grains with methanol as a supplemented carbon source. The biofilms are quite irregular, having a median thickness of 13 to 20 microns. Fatty acid analysis of 56 den...

  6. Whole-Genome Transcriptional Analysis of Chemolithoautotrophic Thiosulfate Oxidation by Thiobacillus denitrificans under Aerobic versus Denitrifying Conditions†

    Beller, Harry R.; Letain, Tracy E.; Chakicherla, Anu; Kane, Staci R.; Legler, Tina C.; Coleman, Matthew A.

    2006-01-01

    Thiobacillus denitrificans is one of the few known obligate chemolithoautotrophic bacteria capable of energetically coupling thiosulfate oxidation to denitrification as well as aerobic respiration. As very little is known about the differential expression of genes associated with key chemolithoautotrophic functions (such as sulfur compound oxidation and CO2 fixation) under aerobic versus denitrifying conditions, we conducted whole-genome, cDNA microarray studies to explore this topic systemat...

  7. Seasonal variations of nitrate reducing and denitrifying bacteria utilizing hexadecane in Mandovi estuary, Goa, West Coast of India

    Sousa, T.D.; Ingole, B.; Sousa, S.D.; Bhosle, S.

    Journal of Geo-Marine Sciences Vol. 42(5), September 2013, pp. 587-592 Seasonal variations of nitrate reducing and denitrifying bacteria utilizing hexadecane in Mandovi estuary, Goa, West Coast of India Trelita de Sousa1, Baban Ingole2...-monsoon) from 4 stations (Fig. 1): Panjim ________ *Corresponding author INDIAN J. MAR. SCI., VOL. 42, NO. 5, SEPTEMBER 2013 588 jetty, Panjim (A); Mangrove ecosystem in the backwaters of River Mandovi, Merces (B); Ribandar jetty, Ribandar (C); Mangrove...

  8. Toxic effects exerted on methanogenic, nitrifying and denitrifying bacteria by chemicals used in a milk analysis laboratory

    Lopez-Fiuza, J.; Buys, B.; Mosquera-Corral, A.; Omil, F.; Mendez, R.

    2002-01-01

    The toxic effects caused by the chemicals contained in wastewaters generated by laboratories involved in raw milk analyses were assessed using batch assays. These assays were carried out separately with methanogenic, ammonium-oxidizing, nitrite-oxidizing and denitrifying bacteria. Since sodium azide is one of the main components of the chemical mixture present in these streams, a set of assays was carried out with the whole chemical mixture, and another one was performed only with azide as th...

  9. Feasibility of enhancing the DEnitrifying AMmonium OXidation (DEAMOX) process for nitrogen removal by seeding partial denitrification sludge.

    Cao, Shenbin; Peng, Yongzhen; Du, Rui; Wang, Shuying

    2016-04-01

    The recently proposed DEnitrifying AMmonium OXidation (DEAMOX) process combined anaerobic ammonia oxidation (ANAMMOX) with denitrification to convert nitrate to nitrite, which was a promising way for treating wastewater containing nitrate and ammonia. This study investigated the feasibility of establishing DEAMOX process by seeding partial denitrification sludge (NO3(-) → NO2(-)) using sodium acetate as an electron donor in a sequencing batch reactor. Results showed that the DEAMOX process was established successfully and operated stably in 114-days operation. The average effluent total nitrogen concentration was below 5 mg L(-1) and TN removal efficiency reached up to 97% at COD/NO3(-) ratio of 3.0 under initial NH4(+) concentration of 25 mg L(-1) and NO3(-) of 30 mg L(-1). It suggested that the presence of NO2(-) in the system supplied for ANAMMOX and the relatively long sludge retention time (SRT) for denitrifiers were attributed to commendable coexistence of ANAMMOX and denitrifying bacteria. PMID:26829308

  10. Coexistence of nitrifying, anammox and denitrifying bacteria in a sequencing batch reactor

    MichelaLangone

    2014-02-01

    Full Text Available Elevated nitrogen removal efficiencies from ammonium-rich wastewaters have been demonstrated by several applications, that combine nitritation and anammox processes. Denitrification will occur simultaneously when organic carbon is also present. In this study, the activity of aerobic ammonia oxidizing, anammox and denitrifying bacteria in a full scale Sequencing Batch Reactor, treating digester supernatants, was studied by means of batch-assays. AOB and anammox activities were maximum at pH of 8.0 and 7.8-8.0, rispectively. Short term effect of nitrite on anammox activity was studied, showing nitrite up to 42 mg/L did not result in inhibition. Both denitrification via nitrate and nitrite were measured. To reduce nitrite-oxidizing activity, high of NH3 – N (1.9-10 mg N-NH3/L and low nitrite (3-8 mg TNN/L are required conditions during the whole SBR cycle.Molecular analysis showed the nitritation-anammox sludge harbored a high microbial diversity, where each microorganism has a specific role. Using ammonia monooxygenase α –subunit (amoA gene as a marker, our analyses suggested different macro- and micro-environments in the reactor strongly affect the AOB community, allowing the development of different AOB species, such as N. europaea/eutropha and N. oligotropha groups, which improve the stability of nitritation process. A specific PCR primer set, used to target the 16S rRNA gene of anammox bacteria, confirmed the presence of the “Ca. Brocadia fulgida” type, able to grow in precence of organic matter and to tolerate high nitrite concentrations. The diversity of denitrifiers was assessed by using dissimilatory nitrite reductase (nirS gene-based analyses, who showed denitifiers were related to different betaproteobacterial genera, such as Thauera, Pseudomonas, Dechloromonas and Aromatoleum, able to assist in forming microbial aggregates. Concerning possible secondary processes, no n-damo bacteria were found while NOB from the genus of Nitrobacter was detected.

  11. Oxygen enrichment incineration

    Oxygen enriched combustion technology has recently been used in waste incineration. To apply the oxygen enrichment on alpha-bearing waste incineration, which is being developed, a state-of-an-art review has been performed. The use of oxygen or oxygen-enriched air instead of air in incineration would result in increase of combustion efficiency and capacity, and reduction of off-gas product. Especially, the off-gas could be reduced below a quarter, which might reduce off-gas treatment facilities, and also increase an efficiency of off-gas treatment. However, the use of oxygen might also lead to local overheating and high nitrogen oxides (NOx) formation. To overcome these problems, an application of low NOx oxy-fuel burner and recycling of a part of off-gas to combustion chamber have been suggested

  12. Laser and gas centrifuge enrichment

    Heinonen, Olli [Senior Fellow, Belfer Center for Science and International Affairs, Harvard Kennedy School, Cambridge, Massachusetts (United States)

    2014-05-09

    Principles of uranium isotope enrichment using various laser and gas centrifuge techniques are briefly discussed. Examples on production of high enriched uranium are given. Concerns regarding the possibility of using low end technologies to produce weapons grade uranium are explained. Based on current assessments commercial enrichment services are able to cover the global needs of enriched uranium in the foreseeable future.

  13. Biogeographical distribution of denitrifying anaerobic methane oxidizing bacteria in Chinese wetland ecosystems.

    Zhu, Guibing; Zhou, Leiliu; Wang, Yu; Wang, Shanyun; Guo, Jianhua; Long, Xi-En; Sun, Xingbin; Jiang, Bo; Hou, Qiaoyun; Jetten, Mike S M; Yin, Chengqing

    2015-02-01

    The discovery of denitrifying anaerobic methane oxidation with nitrite as electron acceptor mediated by 'Candidatus?Methylomirabilis oxyfera' connected the biogeochemical carbon and nitrogen cycle in a new way. However, it is important to have a comprehensive understanding about the distribution of M.?oxyfera-like bacteria in the terrestrial realm, especially the wetland ecosystems that are known as the largest natural source of atmospheric methane. Here, our molecular evidence demonstrated that a wide geographical distribution of M.?oxyfera-like bacteria at oxic/anoxic interfaces of various wetlands (n?=?91) over the Chinese territory. Intriguingly, the M.?oxyfera-like bacteria were detected in some extreme environments, indicating that M.?oxyfera-like bacteria occupied a wide range of habitats. Quantitative polymerase chain reaction estimated that the abundance of M.?oxyfera-like bacteria ranged from 2.2??10(3) to 2.3??10(7) copies?g(-1) dry soil, and up to around 0.62% of the total number of bacteria. Moreover, the M.?oxyfera-like bacteria showed high biodiversity in wetland ecosystems based on the analysis of 462 pmoA and 287 16S rRNA gene sequences. The current study revealed the widespread distribution and biogeography of M.?oxyfera-like bacteria in the terrestrial system. PMID:25223900

  14. Advanced uranium enrichment processes

    Three advanced Uranium enrichment processes are dealt with in the report: AVLIS (Atomic Vapour LASER Isotope Separation), MLIS (Molecular LASER Isotope Separation) and PSP (Plasma Separation Process). The description of the physical and technical features of the processes constitutes a major part of the report. If further presents comparisons with existing industrially used enrichment technologies, gives information on actual development programmes and budgets and ends with a chapter on perspectives and conclusions. An extensive bibliography of the relevant open literature is added to the different subjects discussed. The report was drawn up by the nuclear research Centre (CEA) Saclay on behalf of the Commission of the European Communities

  15. Studies on the enrichment culture of hydrocarbon oxidizing bacteria for MEOR. Part 3. ; Effect of adding yeast extract on the function of bacteria. Sekiyu no zokaishu wo mokuteki to shita tanka suiso riyokin no shuseki baiyo ni kansuru kenkyu. 3. ; Kobo ekisu tenka no eikyo

    Kinoshita, A.; Enomoto, H.; Chida, T. (Tohoku University, Sendai (Japan). Faculty of Engineering)

    1994-03-01

    Possibility of minimizing sources of nutritions to be injected and reducing crude oil density may be expected if crude oil in an oil bed can be utilized as a nutrition source for microorganisms in an enhanced oil recovery. With the expectation, studies are being performed on the enrichment culture of hydrocarbon oxidizing bacteria. This paper describes an experiment on the effect of adding yeast extract using four strains, standing on the results reported in up to the previous paper. The result may be summarized as follows: the addition of yeast extract has made proliferation more active, with all of the four strains having their bacterial concentration increased to 10[sup 8] - 10[sup 9] cells/ml in one to two days; decrease has been seen in the increase of specific growth rate (about 4 to 9 times against the reference culture ground), alkalinity in the culture liquid pH at the termination of the culture experiment lasting two weeks, and interface tension between the culture liquid and kerosene; and the method to multiplicate microorganisms by adding a small amount of yeast extract to a place where petroleum exists is judged an effective method for MEOR. 3 refs., 10 figs., 3 tabs.

  16. Everyone Vegetarian, World Enriching

    Wu, John Y.

    2014-01-01

    This essay advocates global vegetarian diet. Firstly, seven personal health benefits and four global benefits of vegetarian dinners are specified to enrich the entire globe, and then, secondly, I explore concretely how to overcome internal hurdle and external hurdle, so as to effectively propagate vegetarian dinners throughout the world. Everyone wins, including animals!

  17. An enriching industrial project

    The TRICASTIN industrial gas-diffusion uranium enrichment plant which supplies fuel to nuclear power stations is discussed. A concise history of the plant is given and this is followed by a discussion on what has been learnt during the development. Financial, commercial, constructional, and multinational aspects are pinpointed. (L.F.)

  18. Promotion of uranium enrichment business

    The Committee on Nuclear Power has studied on the basic nuclear power policy, establishing its five subcommittees, entrusted by the Ministry of Nternational Trade and Industry. The results of examination by the subcommittee on uranium enrichment business are given along with a report in this connection by the Committee. In order to establish the nuclear fuel cycle, the aspect of uranium enrichment is essential. The uranium enrichment by centrifugal process has proceeded steadily in Power Reactor and Nuclear Fuel Development Corporation. The following matters are described: the need for domestic uranium enrichment, the outlook for overseas enrichment services and the schedule for establishing domestic enrichment business, the current state of technology development, the position of the prototype enrichment plant, the course to be taken to establish enrichment business the main organization operating the prototype and commercial plants, the system of supplying centrifuges, the domestic conversion of natural uranium the subsidies for uranium enrichment business. (J.P.N.)

  19. The role of paraffin oil on the interaction between denitrifying anaerobic methane oxidation and Anammox processes.

    Fu, Liang; Ding, Zhao-Wei; Ding, Jing; Zhang, Fang; Zeng, Raymond J

    2015-10-01

    Methane is sparingly soluble in water, resulting in a slow reaction rate in the denitrifying anaerobic methane oxidation (DAMO) process. The slow rate limits the feasibility of research to examine the interaction between the DAMO and the anaerobic ammonium oxidation (Anammox) process. In this study, optimized 5 % (v/v) paraffin oil was added as a second liquid phase to improve methane solubility in a reactor containing DAMO and Anammox microbes. After just addition, methane solubility was found to increase by 25 % and DAMO activity was enhanced. After a 100-day cultivation, the paraffin reactor showed almost two times higher consumption rates of NO3 (-) (0.2268 mmol/day) and NH4 (+) (0.1403 mmol/day), compared to the control reactor without paraffin oil. The microbes tended to distribute in the oil-water interface. The quantitative (q) PCR result showed the abundance of gene copies of DAMO archaea, DAMO bacteria, and Anammox bacteria in the paraffin reactor were higher than those in the control reactor after 1 month. Fluorescence in situ hybridization revealed that the percentages of the three microbes were 55.5 and 77.6 % in the control and paraffin reactors after 100 days, respectively. A simple model of mass balance was developed to describe the interactions between DAMO and Anammox microbes and validate the activity results. A mechanism was proposed to describe the possible way that paraffin oil enhanced DAMO activity. It is quite clear that paraffin oil enhances not only DAMO activity but also Anammox activity via the interaction between them; both NO3 (-) and NH4 (+) consumption rates were about two times those of the control. PMID:26036704

  20. Nitrate reduction by denitrifying anaerobic methane oxidizing microorganisms can reach a practically useful rate.

    Cai, Chen; Hu, Shihu; Guo, Jianhua; Shi, Ying; Xie, Guo-Jun; Yuan, Zhiguo

    2015-12-15

    Methane in biogas has been proposed to be an electron donor to facilitate complete nitrogen removal using denitrifying anaerobic methane oxidizing (DAMO) microorganisms in an anaerobic ammonium oxidation (anammox) reactor, by reducing the nitrate produced. However, the slow growth and the low activity of DAMO microorganisms cast a serious doubt about the practical usefulness of such a process. In this study, a previously established lab-scale membrane biofilm reactor (MBfR), with biofilms consisting of a coculture of DAMO and anammox microorganisms, was operated to answer if the DAMO reactor can achieve a nitrate reduction rate that can potentially be applied for wastewater treatment. Through progressively increasing nitrate and ammonium loading rates to the reactor, a nitrate removal rate of 684 ± 10 mg-N L(-1) d(-1) was achieved after 453 days of operation. This rate is, to our knowledge, by far the highest reported for DAMO reactors, and far exceeds what is predicted to be required for nitrate removal in a sidestream (5.6-135 mg-N L(-1) d(-1)) or mainstream anammox reactor (3.2-124 mg-N L(-1) d(-1)). Mass balance analysis showed that the nitrite produced by nitrate reduction was jointly reduced by anammox bacteria at a rate of 354 ± 3 mg-N L(-1) d(-1), accompanied by an ammonium removal rate of 268 ± 2 mg-N L(-1) d(-1), and DAMO bacteria at a rate of 330 ± 9 mg-N L(-1) d(-1). This study shows that the nitrate reduction rate achieved by the DAMO process can be high enough for removing nitrate produced by anammox process, which would enable complete nitrogen removal from wastewater. PMID:26414889

  1. Simultaneous electricity generation and pollutant removal in microbial fuel cell with denitrifying biocathode over nitrite

    Highlights: • An MFC was successfully started up using nitrite as cathodic electron acceptor. • The optimal HRT was deemed to be 8 h in this study. • The suitable temperature for power generation was found to be 20 °C. • The suitable temperature for pollutant removal was found to be 25 °C. • Free buffer led to 50% decrease of TN removal rate and power generation. - Abstract: The influences of hydraulic retention time, temperature and free buffer on the performance of short-cut denitrifying microbial fuel cell were investigated after it was successfully started up using nitrite as the cathodic electron acceptor. The results revealed that a power density of 8.3 ± 0.5 W m−3 NC was obtained after 15 days operation. The desirable hydraulic retention time was found in this study to be 8 h, with a COD removal rate of 2.117 ± 0.006 kg m−3 NC d−1 and a total nitrogen removal rate of 0.041 ± 0.002 kg m−3 NC d−1, respectively. It demonstrated that temperature had different effects on the electricity generation and pollutant removal performance of microbial fuel cell. The suitable temperature for power generation and pollutant removal was found to be 20 °C and 25 °C, respectively. Free buffer led to 50% decrease of both total nitrogen removal rate and power density of microbial fuel cell compared to that with phosphate buffer solution addition. The optimal total nitrogen removal rate obtained in the case with sodium azide addition (0.075 ± 0.008 kg m−3 NC d−1) increased by 50% as compared to that without sodium azide addition. It suggested that abolishing oxygen or inhibiting nitrite oxidizing bacteria would favor nitrogen removal

  2. Decaying cyanobacteria decrease N2O emissions related to diversity of intestinal denitrifiers of Chironomus plumosus

    Xu Sun

    2014-10-01

    Full Text Available Nitrous oxide (N2O emission of fresh invertebrates has too long been neglected in eutrophic lakes, although the sediments these animals inhabit are presumably hot spots of N2O emission. Thus, the experiment in this research was designed to gain insight into the influence of cyanobacterial degradation on the N2O emission by fresh water invertebrates (Chironomus plumosus. The presence of decaying cyanobacteria in Lake Taihu decreased the N2O emission rate of Chironomus plumosus larvae from the larvae body by almost 400% for the larvae as a whole. The N2O emission rate decreased by 350% based on readings from studies of their gut, which was mostly due to stimulation of intestinal complete denitrification. The quantitative PCR results showed that intestinal gene abundance of nirK, nosZ (encoding the copper nitrite reductase and N2O reductase, respectively were significantly increased with the presence of decaying cyanobacteria. In contrast nirS (encoding the cytochrome cd1 heme nitrite reductase and the total bacteria decreased. In the gut of Chironomus plumosus, the diversity and richness of nosZ and nirK were lower with the cyanobacteria. Phylogenetic analysis of the intestinal function genes (nosZ and nirK showed that the nosZ- and nirK-type denitrifying bacterial sequences were related to different phylotypes. Hence, additional cyanobacteria increased the abundance, but decreased the richness and diversity of intestinal nitrate-reducing bacteria, probably by providing more carbon source in the gut. The data obtained in this study elucidates that the decaying cyanobacteria decreased the emissions of N2O by the aquatic invertebrates in freshwater sediment and could serve as a valuable resource for nitrogen removal affecting greenhouse gas emissions.

  3. Denitrifying and diazotrophic community responses to artificial warming in permafrost and tallgrass prairie soils.

    Penton, Christopher R; St Louis, Derek; Pham, Amanda; Cole, James R; Wu, Liyou; Luo, Yiqi; Schuur, E A G; Zhou, Jizhong; Tiedje, James M

    2015-01-01

    Increasing temperatures have been shown to impact soil biogeochemical processes, although the corresponding changes to the underlying microbial functional communities are not well understood. Alterations in the nitrogen (N) cycling functional component are particularly important as N availability can affect microbial decomposition rates of soil organic matter and influence plant productivity. To assess changes in the microbial component responsible for these changes, the composition of the N-fixing (nifH), and denitrifying (nirS, nirK, nosZ) soil microbial communities was assessed by targeted pyrosequencing of functional genes involved in N cycling in two major biomes where the experimental effect of climate warming is under investigation, a tallgrass prairie in Oklahoma (OK) and the active layer above permafrost in Alaska (AK). Raw reads were processed for quality, translated with frameshift correction, and a total of 313,842 amino acid sequences were clustered and linked to a nearest neighbor using reference datasets. The number of OTUs recovered ranged from 231 (NifH) to 862 (NirK). The N functional microbial communities of the prairie, which had experienced a decade of experimental warming were the most affected with changes in the richness and/or overall structure of NifH, NirS, NirK and NosZ. In contrast, the AK permafrost communities, which had experienced only 1 year of warming, showed decreased richness and a structural change only with the nirK-harboring bacterial community. A highly divergent nirK-harboring bacterial community was identified in the permafrost soils, suggesting much novelty, while other N functional communities exhibited similar relatedness to the reference databases, regardless of site. Prairie and permafrost soils also harbored highly divergent communities due mostly to differing major populations. PMID:26284038

  4. Optimizing Hydraulic Retention Times in Denitrifying Woodchip Bioreactors Treating Recirculating Aquaculture System Wastewater.

    Lepine, Christine; Christianson, Laura; Sharrer, Kata; Summerfelt, Steven

    2016-05-01

    The performance of wood-based denitrifying bioreactors to treat high-nitrate wastewaters from aquaculture systems has not previously been demonstrated. Four pilot-scale woodchip bioreactors (approximately 1:10 scale) were constructed and operated for 268 d to determine the optimal range of design hydraulic retention times (HRTs) for nitrate removal. The bioreactors were operated under HRTs ranging from 6.6 to 55 h with influent nitrate concentrations generally between 20 and 80 mg NO-N L. These combinations resulted in N removal rates >39 g N m d, which is greater than previously reported. These high removal rates were due in large part to the relatively high chemical oxygen demand and warm temperature (∼19°C) of the wastewater. An optimized design HRT may not be the same based on metrics of N removal rate versus N removal efficiency; longer HRTs demonstrated higher removal efficiencies, and shorter HRTs had higher removal rates. When nitrate influent concentrations were approximately 75 mg NO-N L ( = 6 sample events), the shortest HRT (12 h) had the lowest removal efficiency (45%) but a significantly greater removal rate than the two longest HRTs (42 and 55 h), which were N limited. Sulfate reduction was also observed under highly reduced conditions and was exacerbated under prolonged N-limited environments. Balancing the removal rate and removal efficiency for this water chemistry with a design HRT of approximately 24 h would result in a 65% removal efficiency and removal rates of at least 18 g N m d. PMID:27136146

  5. Different response of nitrifiers and denitrifiers to re-wetting shape the NO release from soils in laboratory incubation experiments

    Behrendt, Thomas; Wu, Dianming; Song, Guozheng; Pommerenke, Bianca; Braker, Gesche

    2014-05-01

    Laboratory incubation studies incubating soils at a wide range of soil moistures, soil temperatures and mixing ratios of NO in the headspace and inorganic nutrient contents (NH4+,NO3-, NO2-) showed that release rates of NO follow an exponential function with increasing soil temperature and an optimum function for soil moisture. This approach assumes that environmental factors play the dominant role in shaping an ecosystem and thereby microbial communities and control the NO release from soil. We determined the NO release rate for a dryland farming soil under flooding irrigation in Xinjiang, China, a mid-latitude agricultural soil (Mainz, Germany), and a rice paddy (Ambai, India) upon wetting and subsequent drying out of soils. Only the release rate for the mid-latitude agricultural soil followed an optimum function for soil moisture. Release rates for the dryland farming soil and rice paddy, however, followed a two maxima function with distinct maxima at higher and lower soil moisture. Acknowledgement of two distinct maxima is critical for more accurately assessing regional biogenic NO emissions of soils under field conditions. To analyse the response of nitrifiers and denitrifiers involved in NO turnover in more detail we linked molecular analysis of functional gene expression (nirK and nirS, bacterial and archaeal amoA) and microbial community composition to NO release rates. We could show that the maximal transcriptional activity of denitrifiers and ammonia oxidizers differs with soil moisture and that higher transcriptional activity of nirS-type denitrifiers at higher soil moisture and of archaeal ammonia oxidizers at lower soil moisture may explain the two maxima for NO release.

  6. Denitrifiers in the surface zone are primarily responsible for the nitrous oxide emission of dairy manure compost

    Highlights: ► Nitrous oxide (N2O) productions of each compost zones were compared. ► The pile surface emitted significant fluxes of N2O. ► The isotopic signature of N2O from surface and NO2− amended core were different. ► The denitrifying gene abundance was significantly higher in pile surface than the pile core. -- Abstract: During the dairy manure composting process, significant nitrous oxide (N2O) emissions occur just after the pile turnings. To understand the characteristics of this N2O emission, samples were taken from the compost surface and core independently, and the N2O production was monitored in laboratory incubation experiments. Equal amounts of surface and core samples were mixed to simulate the turning, and the 15N isotope ratios within the molecules of produced N2O were analyzed by isotopomer analysis. The results showed that the surface samples emitted significant levels of N2O, and these emissions were correlated with NOx−-N accumulation. Moreover, the surface samples and surface-core mixed samples incubated at 30 °C produced N2O with a low site preference (SP) value (−0.9 to 7.0‰) that was close to bacteria denitrification (0‰), indicating that denitrifiers in the surface samples are responsible for this N2O production. On the other hand, N2O produced by NO2−-amended core samples and surface samples incubated at 60 °C showed unrecognized isotopic signatures (SP = 11.4–20.3‰). From these results, it was revealed that the N2O production occurring just after the turnings was mainly derived from bacterial denitrification (including nitrifier denitrification) of NOx−-N under mesophilic conditions, and surface denitrifying bacteria appeared to be the main contributor to this process

  7. Enrichment Strategies in Phosphoproteomics.

    Leitner, Alexander

    2016-01-01

    The comprehensive study of the phosphoproteome is heavily dependent on appropriate enrichment strategies that are most often, but not exclusively, carried out on the peptide level. In this chapter, I give an overview of the most widely used techniques. In addition to dedicated antibodies, phosphopeptides are enriched by their selective interaction with metals in the form of chelated metal ions or metal oxides. The negative charge of the phosphate group is also exploited in a variety of chromatographic fractionation methods that include different types of ion exchange chromatography, hydrophilic interaction chromatography (HILIC), and electrostatic repulsion HILIC (ERLIC) chromatography. Selected examples from the literature will demonstrate how a combination of these techniques with current high-performance mass spectrometry enables the identification of thousands of phosphorylation sites from various sample types. PMID:26584921

  8. Future of uranium enrichment

    The increasing amount of separative work being done in government facilities to produce low-enriched uranium fuel for nuclear utilities again raises the question: should this business-type, industrial function be burned over the private industry. The idea is being looked at by the Reagan administration, but faces problems of national security as well as from the unique nature of the business. This article suggests that a joint government-private venture combining enriching, reprocessing, and waste disposal could be the answer. Further, a separate entity using advanced laser technology to deplete existing uranium tails and lease them for fertile blankets in breeder reactors might earn substantial revenues to help reduce the national debt

  9. South Australia, uranium enrichment

    The Report sets out the salient data relating to the establishment of a uranium processing centre at Redcliff in South Australia. It is conceived as a major development project for the Commonwealth, the South Australian Government and Australian Industry comprising the refining and enrichment of uranium produced from Australian mines. Using the data currently available in respect of markets, demand, technology and possible financial return from overseas sales, the project could be initiated immediately with hexafluoride production, followed rapidly in stages by enrichment production using the centrifuge process. A conceptual development plan is presented, involving a growth pattern that would be closely synchronised with the mining and production of yellowcake. The proposed development is presented in the form of an eight-and-half-year programme. Costs in this Report are based on 1975 values, unless otherwise stated. (Author)

  10. Denitrifier communities in tank bromeliads and prospected N2O emissions from tank substrate upon increasing N-deposition

    Suleiman, Marcel; Brandt, Franziska; Brenzinger, Kristof; Martinson, Guntars; Braker, Gesche

    2014-05-01

    It is well known that tropical rainforest soils with total emissions of 1.34 Tg N/yr from the tropics, play a significant role in the global N2O emissions scenarios. Significant contributions were reported particularly for tropical rainforest soils in South and Central America due to the large areas covered by rainforest in this region. In tropical rainforests of the Americas tank bromeliads constitute a prominent group of plants and were shown to significantly contribute to the production of the greenhouse gas methane from tropical forests. It is, however, essentially unknown whether and how bromeliads may contribute to the production of N2O, another important greenhouse gas. It is also unknown whether N2O emissions relate to atmospheric N-deposition and whether an increase in emissions is to be expected upon the prospected increase in N-deposition. We studied the propensity of tank substrate of the bromeliad Werauhia gladioliflora to emit N2O and how this potential is related to the underlying denitrifier communities. In tropical forests of Costa Rica Werauhia gladioliflora is very abundant with 9.85 specimen m-2. Incubation of the tank substrate with increasing amounts of fertilizer to reflect predicted N-deposition scenarios resulted in proportionally increasing net N2O production. Based on the abundance of Werauhia gladioliflora we estimated annual emissions of 395 µg N2O-N m-2 day-1 for N-deposition levels to date which is in the range of tropical soils. At a surplus of N 70% of N2O produced were not reduced leading to accumulation of N2O which agreed well with the finding that 95% of the denitrifiers detected lacked a gene encoding a N2O-reductase and are therefore unable to reduce N2O to dinitrogen. Generally, denitrifiers were highly abundant and ready to denitrify immediately after provision of a nitrogen source because carbon is non-limiting in tank substrate. Our results suggest that tank bromeliad substrate may be a significant source of N2O in neotropical forests and that with prospected increasing future N-depositions annual N2O emissions might increase.

  11. Novel Denitrifying Bacterium Ochrobactrum anthropi YD50.2 Tolerates High Levels of Reactive Nitrogen Oxides▿ †

    Doi, Yuki; Takaya, Naoki; Takizawa, Noboru

    2009-01-01

    Most studies of bacterial denitrification have used nitrate (NO3−) as the first electron acceptor, whereas relatively less is understood about nitrite (NO2−) denitrification. We isolated novel bacteria that proliferated in the presence of high levels of NO2− (72 mM). Strain YD50.2, among several isolates, was taxonomically positioned within the α subclass of Proteobacteria and identified as Ochrobactrum anthropi YD50.2. This strain denitrified NO2−, as well as NO3−. The gene clusters for deni...

  12. Microbial Enrichment of Vermicompost

    Kuppuraj Rajasekar; Thilagavathy Daniel; Natchimuthu Karmegam

    2012-01-01

    The present study has been conducted to explore the possibility of enrichment of vermicompost with microbial inoculants (i.e., biofertilizer organisms), Azospirillum brasilense and Rhizobium leguminosarum, optimization of inoculum level, and time of inoculation during vermicomposting. The survival rate of each microbial inoculant, total microbial population in vermicompost, and their correlation with the microbial inoculants during the storage period (180 days) were assessed. The change in po...

  13. ENRICHMENT OF OBO ONTOLOGIES

    Bada, Michael; Hunter, Lawrence

    2006-01-01

    This paper describes a frame-based integration of the three GO subontologies, the Chemicals of Biological Interest ontology (ChEBI), and the Cell Type Ontology (CTO) in which relationships between elements of the ontologies are modeled in a way that better captures the relational semantics between biological concepts represented by the terms, rather than between the terms themselves, than previous frame-based efforts. We also describe a methodology for creating suggested enriching assertions ...

  14. What is Enrichment For (?)

    Matsangos, P.

    2011-01-01

    It has long been held that schooling, the curriculum (hidden or otherwise), and the pedagogical practices pertaining to them are the perpetrators of social and economic inequalities in Western society. My interest surrounds the role of ‘enrichment’, as an auxiliary of education, in perpetuating or restricting social justice. As the enrichment coordinator within an inner city Sixth Form college, it became part of my remit to research the dispositions held by our largely underprivileged student...

  15. Uranium enrichment technologies

    Several technically mature and commercially proven methods of separation are currently available on the enrichment market, such as the diffusion and centrifuge methods. According to the information available, the diffusion technologies, which have dominated the scene so far, will lose much of their influence in favor of the centrifuge technologies which, because of their special process characteristics, such as lower energy consumption and smaller economic plant sizes, can adapt themselves to the conditions of the market in a flexible and economic way. Other methods, such as the gas dynamics Helikon and separation nozzle techniques, and chemical enrichment are on the verge of commercial maturity. However, their economic performance and chances in the enrichment market cannot yet be assessed with any degree of precision. The prospects and possibilities of the laser method, which has received much backing recently, especially in the USA, are difficult to weigh in the light of the knowledge available to date. However, it is hardly to be expected that technically mature and commercially viable plants operating by this technology are going to be available before the turn of the century. (orig.)

  16. INFLUENCE OF ALTERNATIVE ELECTRON ACCEPTORS ON THE ANAEROBIC BIODEGRADABILITY OF CHLORINATED PHENOLS AND BENZOIC ACIDS

    Nitrate, sulfate, and carbonate were used as electron acceptors to examine the anaerobic biodegradability of chlorinated aromatic compounds in estuarine and freshwater sediments. he respective denitrifying, sulfidogenic, and methanogenic enrichment cultures were established on ea...

  17. pH-driven shifts in overall and transcriptionally active denitrifiers control gaseous product stoichiometry in growth experiments with extracted bacteria from soil

    Brenzinger, Kristof; Dörsch, Peter; Braker, Gesche

    2015-01-01

    Soil pH is a strong regulator for activity as well as for size and composition of denitrifier communities. Low pH not only lowers overall denitrification rates but also influences denitrification kinetics and gaseous product stoichiometry. N2O reductase is particularly sensitive to low pH which seems to impair its activity post-transcriptionally, leading to higher net N2O production. Little is known about how complex soil denitrifier communities respond to pH change and whether their ability ...

  18. New technology for uranium enrichment

    The author reviews new enrichment technologies from the perspective of their impact upon the availability, economics, and security of supply of enriched uranium. Two very attractive enrichment technologies, advanced gas centrifugation and atomic vapour laser isotope separation, are available to the United States. Both should be fully demonstrated in the mid-1980s and should reduce U.S. SWU prices considerably to about $50 or less by the year 2000. World enrichment supplies are adequate to meet demand

  19. Effect of cultivation methods on nutritional enrichment of euryhaline rotifer Brachionus plicatilis

    Kotani, T.; Genka, T.; Fushimi, H.; Hayashi, M.; Dierckens, K.; Sorgeloos, P

    2009-01-01

    This study aimed at comparing fatty acid contents of rotifers cultured with different methods after nutritional enrichment in order to evaluate the rotifer quality produced by these methods. Rotifers were cultured using either a batch or a continuous culture. From the batch culture, three experimental subpopulations were used, sampled from the culture at 1, 24, and 48 h after rotifer inoculation. The continuous culture was performed with two tanks; one was for cultivation with continuous feed...

  20. Characterization of a murine neuron-enriched model of primary trigeminal ganglia cultures to study the interferon- ? antiviral effect against Herpes Simplex Virus type 1 / Caracterizacin de un modelo murino de cultivos primarios de ganglio trigeminal enriquecido en neuronas para el estudio del efecto antiviral del interfern- ? frente al virus Herpes simple tipo 1

    Ana Maria, Low-Calle; Jeanette, Prada-Arismendy; Jaime E., Castellanos.

    2014-06-01

    Full Text Available Objetivo: Caracterizar un cultivo primario de ganglio trigeminal (GT) enriquecido en neuronas y evaluar la expresin de interfern- y su actividad frente a la infeccin con Herpes simple tipo 1 (HSV-1). Materiales y mtodos: El porcentaje de neuronas fue determinado por inmunocitoqumica para neurof [...] ilamento. Los cultivos fueron tratados con interfern- ? e infectados con HSV-1, y se cuantificaron las clulas positivas para antgeno viral por inmunocitoqumica y la expresin de interfern- ? por PCR cuantitativa. Resultados: El cultivo present un 15% de neuronas y 85% de clulas no neuronales. Se encontr efecto citoptico, asociado a una alta diseminacin de la infeccin (72,9% neuronas y 48,3% de clulas no neuronales positivas para antgeno viral). El interfern- ? evit la aparicin de efecto citoptico y disminuy las clulas infectadas a 16,7% en neuronas y a 7,8% las clulas no neuronales. La infeccin viral increment la expresin de transcritos de interfern- ? 18,2 veces a las 6 h de infeccin, mientras que a las 18 h post infeccin el tratamiento con interfern increment esta expresin 3,7 veces. Discusin: Los cultivos presentaron un 15% de neuronas, lo cual es 10 veces ms que en otros cultivos reportados. Las clulas no neuronales representan el 85% de las clulas del cultivo, y se evidenci que todos los tipos de clulas se infectaron; similar a lo que ha sido reportado durante infecciones agudas in vivo . Adicionalmente, el interfern- ? disminuy el porcentaje de clulas infectadas y evit la aparicin de efecto citoptico, similar a lo que ha sido reportado en cultivos de GT porcino. Conclusiones: Se caracteriz un modelo de cultivo primario de GT enriquecido en neuronas. Interfern- ? protegi las clulas del efecto citoptico y la diseminacin viral mientras que la infeccin viral increment la expresin de interfern- ?. Por lo tanto, el interfern- ? ejerci un papel antiviral importante frente al HSV-1 en estos cultivos. Abstract in english Objective: To characterize a neuron-enriched primary TG culture and evaluate interferon- ? expression and activity after HSV-1 infection. Materials and methods: The percentage of neurons present in cultures was assessed by neurofilament immunocytochemistry. Cultures were treated with interferon- ? a [...] nd infected with HSV-1, then viral antigen positive cells were counted and interferon- ?expression was assessed by quantitative PCR. Results: The culture contained 15% neurons and 85% non-neuronal cells. A cytopathic effect was observed, associated with high viral spread (72.9% neurons and 48.3% non-neuronal cells were positive for viral antigen). Interferon- ? treatment impaired the cytopathic effect and decreased the infected neurons to 16.7% and infected non-neuronal cells to 7.8%. Viral infection at 6 h postinfection significantly increased the interferon- ? transcripts by 18.2 fold, while at 18 h postinfection Interferon pre-treatment in infected cultures increased interferon- ? transcription by 3.7 fold. Discussion: This culture model contained 15% neurons, which is 10 times higher compared to other reported cultures, and non-neuronal cells comprised 85% of cells in this culture. All types of cells were found to be infected, which is similar to that reported during acute infections in vivo . Additionally, interferon- ?decreased the infected cells, avoiding the cytopathic effect, which is similar to that reported in swine TG cultures. Conclusions: A neuron-enriched primary TG model was characterized. Interferon- ? treatment protected cells from cytopathic effects and viral spread, while viral infection up-regulated interferon- ? expression. This result means that interferon- ? exerts an important antiviral effect against HSV-1 in these cultures.

  1. Estimate of denitrifying microbiota in tertiary sewage treatment and kinetics of the denitrification process using different sources of carbon

    Marchetto Margarida

    2003-01-01

    Full Text Available A study of the kinetics of denitrification was carried out in the laboratory based on the quantification of N2O, the final product of the activity of denitrifying microorganisms, when the enzymatic reduction of N2O to N2 was blocked by acetylene. Concentrated mixed liquor (sludge from a reactor with intermittent aeration used for sewage treatment was used as the inoculum, while methanol, acetic acid, glucose, effluent sewage from an anaerobic fluidized bed reactor and synthetic substrate simulating domestic sewage were used as carbon sources. The mean concentration of nitrate was 20 mg/L. Maxima of N2O production and NO3- consumption occurred between 0.5h and 2.0h of incubation using all the carbon sources, which characterized the denitrification process. Acetic acid and methanol were responsible for the highest rates of N2O production. The estimated number of denitrifying microorganisms in the reactor with intermittent aeration, using the MPN technique, varied from 10(9 to 10(10 MPN/g VSS, indicating a high potential for the occurrence of denitrification.

  2. Denitrifying bacteria from the genus Rhodanobacter dominate bacterial communities in the highly contaminated subsurface of a nuclear legacy waste site

    Green, Stefan [Florida State University; Prakash, Om [Florida State University; Jasrotia, Puja [Florida State University; Overholt, Will [Florida State University; Cardenas, Erick [Michigan State University, East Lansing; Hubbard, Daniela [Florida State University; Tiedje, James M. [Michigan State University, East Lansing; Watson, David B [ORNL; Schadt, Christopher Warren [ORNL; Brooks, Scott C [ORNL; Kostka, Joel [Florida State University

    2011-01-01

    The effect of long-term mixed-waste contamination, particularly uranium and nitrate, on the microbial community in the terrestrial subsurface was investigated at the field scale at the Oak Ridge Integrated Field Research Challenge (ORIFRC) site in Oak Ridge, TN. The abundance, community composition, and distribution of groundwater microorganisms were examined across the site during two seasonal sampling events. At representative locations, subsurface sediment was also examined from two boreholes, one sampled from the most heavily contaminated area of the site and another from an area with low contamination. A suite of DNA- and RNA-based molecular tools were employed for community characterization, including quantitative PCR of ribosomal RNA and nitrite reductase genes, community composition fingerprinting analysis, and high-throughput pyrotag sequencing of rRNA genes. The results demonstrate that pH is a major driver of the subsurface microbial community structure, and denitrifying bacteria from the genus Rhodanobacter (class Gammaproteobacteria) dominate at low pH. The relative abundance of bacteria from this genus was positively correlated with lower pH conditions, and these bacteria were abundant and active in the most highly contaminated areas. Other factors, such as concentration of nitrogen species, oxygen and sampling season did not appear to strongly influence the distribution of Rhodanobacter. Results indicate that these organisms are acid-tolerant denitrifiers, well suited to the acidic, nitrate-rich subsurface conditions, and pH is confirmed as a dominant driver of bacterial community structure in this contaminated subsurface environment.

  3. Isolation of the ε-caprolactam denitrifying bacteria from a wastewater treatment system manufactured with acrylonitrile-butadiene-styrene resin

    ε-Caprolactam has high COD and toxicity, so its discharge to natural water and soil systems may lead to an adverse environmental effect on water quality, endangering public health and welfare. This investigation attempts to isolate ε-caprolactam denitrifying bacteria from a wastewater treatment system manufactured with acrylonitrile-butadiene-styrene (ABS) resin. The goal is to elucidate the effectiveness of isolated pure strain and ABS mixed strains in treating ε-caprolactam from synthetic wastewater. The results reveal that Paracoccus versutus MDC-3 was isolated from the wastewater treatment system manufactured with ABS resin. The ABS mixed strains and P. versutus MDC-3 can consume up to 1539 mg/l ε-caprolactam to denitrify from synthetic wastewater. Complete ε-caprolactam removal depended on the supply of sufficient electron acceptors (nitrate). Strain P. versutus MDC-3, Hyphomicrobium sp. HM, Methylosinus pucelana and Magnetospirillum sp. CC-26 are related closely, according to the phylogenetic analyses of 16S rDNA sequences

  4. Effect of the C:N:P ratio on the denitrifying dephosphatation in a sequencing batch biofilm reactor (SBBR).

    Mielcarek, Artur; Rodziewicz, Joanna; Janczukowicz, Wojciech; Thornton, Arthur J; Jóźwiak, Tomasz; Szymczyk, Paula

    2015-12-01

    A series of investigations were conducted using sequencing batch biofilm reactor (SBBR) to explore the influence of C:N:P ratio on biological dephosphatation including the denitrifying dephosphatation and the denitrification process. Biomass in the reactor occurred mainly in the form of a biofilm attached to completely submerged disks. Acetic acid was used as the source of organic carbon. C:N:P ratios have had a significant effect on the profiles of phosphate release and phosphate uptake and nitrogen removal. The highest rates of phosphate release and phosphate uptake were recorded at the C:N:P ratio of 140:70:7. The C:N ratio of 2.5:1 ensured complete denitrification. The highest rate of denitrification was achieved at the C:N:P ratio of 140:35:7. The increase of nitrogen load caused an increase in phosphates removal until a ratio C:N:P of 140:140:7. Bacteria of the biofilm exposed to alternate conditions of mixing and aeration exhibited enhanced intracellular accumulation of polyphosphates. Also, the structure of the biofilm encouraged anaerobic-aerobic as well as anoxic-anaerobic and absolutely anaerobic conditions in a SBBR. These heterogeneous conditions in the presence of nitrates may be a significant factor determining the promotion of denitrifying polyphosphate accumulating organism (DNPAO) development. PMID:26702975

  5. Effect of carbon dioxide and bicarbonate as inorganic carbon sources on growth and adaptation of autohydrogenotrophic denitrifying bacteria

    Acclimation of autohydrogenotrophic denitrifying bacteria using inorganic carbon source (CO2 and bicarbonate) and hydrogen gas as electron donor was performed in this study. In this regard, activated sludge was used as the seed source and sequencing batch reactor (SBR) technique was applied for accomplishing the acclimatization. Three distinct strategies in feeding of carbon sources were applied: (I) continuous sparging of CO2, (II) bicarbonate plus continuous sparging of CO2, and (III) only bicarbonate. The pH-reducing nature of CO2 showed an unfavorable impact on denitrification rate; however bicarbonate resulted in a buffered environment in the mixed liquor and provided a suitable mean to maintain the pH in the desirable range of 7-8.2. As a result, bicarbonate as the only carbon source showed a faster adaptation, while carbon dioxide as the only carbon source as well as a complementary carbon source added to bicarbonate resulted in longer acclimation period. Adapted hydrogenotrophic denitrifying bacteria, using bicarbonate and hydrogen gas in the aforementioned pH range, caused denitrification at a rate of 13.33 mg NO3--N/g MLVSS/h for degrading 20 and 30 mg NO3--N/L and 9.09 mg NO3--N/g MLVSS/h for degrading 50 mg NO3--N/L

  6. Nitrate-dependent anaerobic ferrous oxidation (NAFO) by denitrifying bacteria: a perspective autotrophic nitrogen pollution control technology.

    Zhang, Meng; Zheng, Ping; Wang, Ru; Li, Wei; Lu, Huifeng; Zhang, Jiqiang

    2014-12-01

    The nitrate-dependent anaerobic ferrous oxidation (NAFO) is an important discovery in the fields of microbiology and geology, which is a valuable biological reaction since it can convert nitrate into nitrogen gas, removing nitrogen from wastewater. The research on NAFO can promote the development of novel autotrophic biotechnologies for nitrogen pollution control and get a deep insight into the biogeochemical cycles. In this work, batch experiments were conducted with denitrifying bacteria as biocatalyst to investigate the performance of nitrogen removal by NAFO. The results showed that the denitrifying bacteria were capable of chemolithotrophic denitrification with ferrous salt as electron donor, namely NAFO. And the maximum nitrate conversion rates (qmax) reached 57.89 mg (g VSS d)−1, which was the rate-limiting step in NAFO. Fe/N ratio, temperature and initial pH had significant influences on nitrogen removal by NAFO process, and their optimal values were 2.0 °C, 30.15 °C and 8.0 °C, respectively. PMID:25461924

  7. Acidophilic denitrifiers dominate the N2O production in a 100-year-old tea orchard soil.

    Huang, Ying; Long, Xi-En; Chapman, Stephen J; Yao, Huaiying

    2015-03-01

    Aerobic denitrification is the main process for high N2O production in acid tea field soil. However, the biological mechanisms for the high emission are not fully understood. In this study, we examined N2O emission and denitrifier communities in 100-year-old tea soils with four pH levels (3.71, 5.11, 6.19, and 7.41) and four nitrate concentration (0, 50, 200, and 1000 mg kg(-1) of NO3 (-)-N) addition. Results showed the highest N2O emission (10.1 mg kg(-1) over 21 days) from the soil at pH 3.71 with 1000 mg kg(-1) NO3 (-) addition. The N2O reduction and denitrification enzyme activity in the acid soils (pH tree analysis. Our results showed that the acidophilic denitrifier adaptation to the acid environment results in high N2O emission in this highly acidic tea soil. PMID:25273518

  8. Preliminary insights into ?15N and ?18O of nitrate in natural mosses: A new application of the denitrifier method

    Natural mosses have been employed as reactive and accumulative indicators of atmospheric pollutants. Using the denitrifier method, the concentration, ?15N and ?18O of moss nitrate (NO3?) were measured to elucidate the sources of NO3? trapped in natural mosses. Oven drying at 5570 C, not lyophilization, was recommended to dry mosses for NO3? analyses. An investigation from urban to mountain sites in western Tokyo suggested that moss [NO3?] can respond to NO3? availability in different habitats. NO3? in terricolous mosses showed isotopic ratios as close to those of soil NO3?, reflecting the utilization of soil NO3?. Isotopic signatures of NO3? in corticolous and epilithic mosses elucidated atmospheric NO3? sources and strength from the urban (vehicle NOx emission) to mountain area (wet-deposition NO3?). However, mechanisms and isotopic effects of moss NO3? utilization must be further verified to enable the application of moss NO3? isotopes for source identification. - Highlights: ? Plant [NO3?] has been used to get insights into forest N saturation. ? However, detailed protocols were not established for plant [NO3?] analysis. ? Moreover, nitrogen/oxygen isotopic ratios of plant NO3? cannot be measured. ? We developed a new protocol for concentration and isotopic measurement of plant NO3?. ? The new method can open a new window to use these parameters for NO3? physiology. - Plant nitrate isotopes can be measured using the denitrifier method and nitrate isotopes of mosses are useful to differentiate atmospheric nitrate sources.

  9. Effect of carbon dioxide and bicarbonate as inorganic carbon sources on growth and adaptation of autohydrogenotrophic denitrifying bacteria

    Ghafari, Shahin; Hasan, Masitah [Department of Chemical Engineering, Faculty of Engineering, University of Malaya, 50603 Kuala Lumpur (Malaysia); Aroua, Mohamed Kheireddine [Department of Chemical Engineering, Faculty of Engineering, University of Malaya, 50603 Kuala Lumpur (Malaysia)], E-mail: mk_aroua@um.edu.my

    2009-03-15

    Acclimation of autohydrogenotrophic denitrifying bacteria using inorganic carbon source (CO{sub 2} and bicarbonate) and hydrogen gas as electron donor was performed in this study. In this regard, activated sludge was used as the seed source and sequencing batch reactor (SBR) technique was applied for accomplishing the acclimatization. Three distinct strategies in feeding of carbon sources were applied: (I) continuous sparging of CO{sub 2}, (II) bicarbonate plus continuous sparging of CO{sub 2}, and (III) only bicarbonate. The pH-reducing nature of CO{sub 2} showed an unfavorable impact on denitrification rate; however bicarbonate resulted in a buffered environment in the mixed liquor and provided a suitable mean to maintain the pH in the desirable range of 7-8.2. As a result, bicarbonate as the only carbon source showed a faster adaptation, while carbon dioxide as the only carbon source as well as a complementary carbon source added to bicarbonate resulted in longer acclimation period. Adapted hydrogenotrophic denitrifying bacteria, using bicarbonate and hydrogen gas in the aforementioned pH range, caused denitrification at a rate of 13.33 mg NO{sub 3}{sup -}-N/g MLVSS/h for degrading 20 and 30 mg NO{sub 3}{sup -}-N/L and 9.09 mg NO{sub 3}{sup -}-N/g MLVSS/h for degrading 50 mg NO{sub 3}{sup -}-N/L.

  10. Uranium enrichment in France

    Eurodif's uranium enrichment plant in France will undoubtedly remain reliable, flexible and competitive for at least the next twenty years. Its replacement is nevertheless already under study and evaluation by Cogema, the main shareholder, since the investment decisions about the technology chosen for the substitution of the gaseous process diffusion will have to be made in near future. Supporting the efforts of the nuclear utilities to keep the nuclear power option viable and competitive, Cogema presently focuses on the development of laser processes, designed to attractively offer fully loaded costs, i.e. including the returns on investment, not much higher than the variable production costs of today. (orig.)

  11. Enriching recycled uranium

    The paper reviews the progress of the use of recycled uranium during the period 1985-8. This article was originally presented as a paper at the 1988 Uranium Institute symposium (which was held in London). A description is given of the differences between natural and recycled uranium, and the presence of U236 in recycled uranium. The concept of equivalent reactivity is described, as well as the cost benefit of using recycled uranium. A summary of Urenco tests and trials with reprocessed uranium is given. Enrichment, UF6 conversion and fuel fabrication are also discussed. (U.K.)

  12. Uranium enrichment by gas centrifuge

    After recalling the physical principles and the techniques of centrifuge enrichment the report describes the centrifuge enrichment programmes of the various countries concerned and compares this technology with other enrichment technologies like gaseous diffusion, laser, aerodynamic devices and chemical processes. The centrifuge enrichment process is said to be able to replace with advantage the existing enrichment facilities in the short and medium term. Future prospects of the process are also described, like recycled uranium enrichment and economic improvements; research and development needs to achieve the economic prospects are also indicated. Finally the report takes note of the positive aspect of centrifuge enrichment as far as safeguards and nuclear safety are concerned. 27 figs, 113 refs

  13. The Cultural Heritage of Interculturalism

    Martin Peterson

    2015-03-01

    Full Text Available The article traces the cultural heritage of inter-cultural contexts, which have had profound impact over long time. It takes its departure in antique and culturally complex environments in the eastern Mediterranean. One millennium later corresponding inter-cultural conditions are explored in the western part of the Mediterranean. Both cases demonstrated their wide and long lasting influences on posterity. The cultural heritage implied the deep effects of cross-fertilization and ensuing cultural enrichment as the conflation of several well-endowed cultures took place. A similar, more powerful outcome followed the Radical Enlightenment in Leiden around 1650s and in Vienna some centuries later.

  14. Cattle grazing and trampling activities affect abundance and activity of nitrifying and denitrifying microbial communities and N.sub.2./sub.O production in upland soil

    Chroňáková, Alica; Radl, V.; Čuhel, Jiří; Gattinger, A.; Šimek, Miloslav; Elhottová, Dana; Schloter, M.

    Braunschweig : Bba - Federal Biological Research Centre for Agriculture and Forestry, 2007. s. 159-160. [Symposium on Bacterial Genetics and Ecology. BAGECO 9. Microbial Community Networks /9./. 23.06.2007-27.06.2007, Wernigerode] Institutional research plan: CEZ:AV0Z60660521 Keywords : cattle grazing and trampling activities * nitrifying and denitrifying microbial communities * upland soil Subject RIV: EH - Ecology, Behaviour

  15. Enrichment of boron 10

    A isotopic separation pilot plant with five ion exchange columns interconnected in series were designed and built in the IEN. The columns are charged with a strong anionic resin in its alkaline form. The boric acid solution is introduced in the separation columns until it reaches a absorbing zone length which is sufficient to obtain the desired boron-10 isotopic concentration. The boric acid absorbing zone movement is provided by the injection of a diluted hydrochloric acid solution, which replace the boric acid throughout the columns. The absorbing zone equilibrium length is proportional to its total length. The enriched boron-10 and the depleted boron are located in the final boundary and in the initial position of the absorbing zones, respectively. (author)

  16. Distribution of typical denitrifying functional genes and diversity of the nirS-encoding bacterial community related to environmental characteristics of river sediments

    Huang, S.; Chen, C.; Yang, X.; Wu, Q.; Zhang, R.

    2011-10-01

    Denitrification in river sediments leads to nitrate removal from the aquatic system; therefore, it is necessary to understand functional diversity of denitrifier communities in the system. Sediment samples (0-25 cm depth) were collected from three typical locations along the Pearl River. The real-time PCR approach was used to measure the abundance of nitrate (narG), nitrite (nirS, nirK and nrfA), and nitrous oxide (nosZ) reductase genes from the sediment samples. Assemblages of nirS, nirK and nosZ indicated that complete denitrification occurred in sediment cores, with the greatest number of gene copies from 5-15 cm depth. Dissimilatory nitrate reduction appeared to be important below 15 cm depth, based on increasing gene copies of narG and nrfA with sediment depth. There was a close match (78-94 %) between the nirS sequences recovered from the Pearl River sediment and those detected in estuarine and marine sediments as well as active sludge, suggesting that the nitrogen source in the Pearl River sediment was affected by domestic sewage inputs and irregular tides. Canonical correspondence analysis indicated that the spatial distribution of denitrifying bacteria was highly correlated with dissolved inorganic nitrogen (including NH4+, NO2- and NO3-) concentrations in sediment. It was concluded that the difference in dissolved inorganic nitrogen concentrations along the sediment profile influenced the distribution of denitrifying genes and the nirS-encoding denitrifier community in the river sediment. In addition, a variety of novel denitrifying bacteria were revealed in the river sediment.

  17. Distribution of typical denitrifying functional genes and diversity of the nirS-encoding bacterial community related to environmental characteristics of river sediments

    R. Zhang

    2011-10-01

    Full Text Available Denitrification in river sediments leads to nitrate removal from the aquatic system; therefore, it is necessary to understand functional diversity of denitrifier communities in the system. Sediment samples (0–25 cm depth were collected from three typical locations along the Pearl River. The real-time PCR approach was used to measure the abundance of nitrate (narG, nitrite (nirS, nirK and nrfA, and nitrous oxide (nosZ reductase genes from the sediment samples. Assemblages of nirS, nirK and nosZ indicated that complete denitrification occurred in sediment cores, with the greatest number of gene copies from 5–15 cm depth. Dissimilatory nitrate reduction appeared to be important below 15 cm depth, based on increasing gene copies of narG and nrfA with sediment depth. There was a close match (78–94 % between the nirS sequences recovered from the Pearl River sediment and those detected in estuarine and marine sediments as well as active sludge, suggesting that the nitrogen source in the Pearl River sediment was affected by domestic sewage inputs and irregular tides. Canonical correspondence analysis indicated that the spatial distribution of denitrifying bacteria was highly correlated with dissolved inorganic nitrogen (including NH4+, NO2− and NO3− concentrations in sediment. It was concluded that the difference in dissolved inorganic nitrogen concentrations along the sediment profile influenced the distribution of denitrifying genes and the nirS-encoding denitrifier community in the river sediment. In addition, a variety of novel denitrifying bacteria were revealed in the river sediment.

  18. Distribution of typical denitrifying functional genes and diversity of the nirS-encoding bacterial community related to environmental characteristics of river sediments

    X. Yang

    2011-05-01

    Full Text Available Denitrification in river sediments leads to nitrate removal from the aquatic system; therefore, it is necessary to understand functional diversity of denitrifier communities in the system. Sediment samples (0–25 cm depth were collected from three typical locations along the Pearl River. The real-time PCR approach was used to measure the abundance of nitrate (narG, nitrite (nirS, nirK and nrfA, and nitrous oxide (nosZ reductase genes from the sediment samples. Assemblages of nirS, nirK and nosZ indicated that complete denitrification occurred in sediment cores, with the greatest number of gene copies from 5–15 cm depth. Dissimilatory nitrate reduction appeared to be important below 15 cm depth, based on increasing gene copies of narG and nrfA with sediment depth. There was a close match (78–94 % between the nirS sequences recovered from Pearl River sediment and those detected in estuarine and marine sediments as well as active sludge, suggesting that domestic sewage inputs and irregular tides. Canonical correspondence analysis indicated that the spatial distribution of denitrifying bacteria was highly correlated with dissolved inorganic N (DIN: NH4+, NO2 and NO3− concentrations in sediment. We conclude that changes in DIN within the sediment profile influences the distribution of denitrifying genes and the nirS-encoding denitrifier community in the river sediment. Our results also reveal a variety of novel denitrifying bacteria in the river sediment.

  19. Advanced Neutron Source enrichment study

    A study has been performed of the impact on performance of using low enriched uranium (20% 235U) or medium enriched uranium (35% 235U) as an alternative fuel for the Advanced Neutron Source, which is currently designed to use uranium enriched to 93% 235U. Higher fuel densities and larger volume cores were evaluated at the lower enrichments in terms of impact on neutron flux, safety, safeguards, technical feasibility, and cost. The feasibility of fabricating uranium silicide fuel at increasing material density was specifically addressed by a panel of international experts on research reactor fuels. The most viable alternative designs for the reactor at lower enrichments were identified and discussed. Several sensitivity analyses were performed to gain an understanding of the performance of the reactor at parametric values of power, fuel density, core volume, and enrichment that were interpolations between the boundary values imposed on the study or extrapolations from known technology

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

    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. PMID:26636960

  1. Denitrifiers in the surface zone are primarily responsible for the nitrous oxide emission of dairy manure compost

    Maeda, Koki, E-mail: k_maeda@affrc.go.jp [Dairy Research Division, National Agricultural Research Center for Hokkaido Region, National Agricultural and Food Research Organization, 1 Hitsujigaoka, Sapporo 062-8555 (Japan); Department of Environmental Chemistry and Engineering, Tokyo Institute of Technology, 4259 Nagatsuta, Midori-ku, Yokohama 226-8502 (Japan); Toyoda, Sakae [Department of Environmental Chemistry and Engineering, Tokyo Institute of Technology, 4259 Nagatsuta, Midori-ku, Yokohama 226-8502 (Japan); Hanajima, Dai [Dairy Research Division, National Agricultural Research Center for Hokkaido Region, National Agricultural and Food Research Organization, 1 Hitsujigaoka, Sapporo 062-8555 (Japan); Yoshida, Naohiro [Department of Environmental Chemistry and Engineering, Tokyo Institute of Technology, 4259 Nagatsuta, Midori-ku, Yokohama 226-8502 (Japan)

    2013-03-15

    Highlights: ► Nitrous oxide (N{sub 2}O) productions of each compost zones were compared. ► The pile surface emitted significant fluxes of N{sub 2}O. ► The isotopic signature of N{sub 2}O from surface and NO{sub 2}{sup −} amended core were different. ► The denitrifying gene abundance was significantly higher in pile surface than the pile core. -- Abstract: During the dairy manure composting process, significant nitrous oxide (N{sub 2}O) emissions occur just after the pile turnings. To understand the characteristics of this N{sub 2}O emission, samples were taken from the compost surface and core independently, and the N{sub 2}O production was monitored in laboratory incubation experiments. Equal amounts of surface and core samples were mixed to simulate the turning, and the {sup 15}N isotope ratios within the molecules of produced N{sub 2}O were analyzed by isotopomer analysis. The results showed that the surface samples emitted significant levels of N{sub 2}O, and these emissions were correlated with NO{sub x}{sup −}-N accumulation. Moreover, the surface samples and surface-core mixed samples incubated at 30 °C produced N{sub 2}O with a low site preference (SP) value (−0.9 to 7.0‰) that was close to bacteria denitrification (0‰), indicating that denitrifiers in the surface samples are responsible for this N{sub 2}O production. On the other hand, N{sub 2}O produced by NO{sub 2}{sup −}-amended core samples and surface samples incubated at 60 °C showed unrecognized isotopic signatures (SP = 11.4–20.3‰). From these results, it was revealed that the N{sub 2}O production occurring just after the turnings was mainly derived from bacterial denitrification (including nitrifier denitrification) of NO{sub x}{sup −}-N under mesophilic conditions, and surface denitrifying bacteria appeared to be the main contributor to this process.

  2. Spectral enrichments of model categories

    Dugger, Daniel

    2005-01-01

    We prove that every stable, combinatorial model category has a natural enrichment by symmetric spectra (or more precisely, a natural equivalence class of enrichments). This in some sense generalizes the simplicial enrichments of model categories provided by the Dwyer-Kan hammock localization. As one particular application, we are able to associate to every object of a stable, combinatorial model category a "homotopy endomorphism ring spectrum". The homotopy types of these ring spectra are pre...

  3. Low enriched fuels for NRU

    Low enriched uranium silicide dispersion fuels are under development for use in Canadian test reactors. These 20% enriched dispersion fuels are to replace the current 93% enriched uranium-aluminum alloy driver fuels. The reduced enrichment is intended to reduce the risk of illegal diversion for weapons proliferation. Developments in uranium silicide dispersion manufacturing technology have proven the production viability of the fuel. Success in the irradiation testing of the dispersion fuels in mini-element form has led to the irradiation of seven full-size fuel assemblies

  4. Do freshwater macrophytes influence the community structure of ammonia-oxidizing and denitrifying bacteria in the rhizospere?

    Herrmann, Martina; Schramm, Andreas

    2006-01-01

    DO FRESHWATER MACROPHYTES INFLUENCE THE COMMUNITY STRUCTURE OF AMMONIA-OXIDIZING AND DENITRIFYING BACTERIA IN THE RHIZOSPHERE? M. Herrmann, A. Schramm Department of Biological Sciences, Microbiology, University of Aarhus, Aarhus, Denmark Aquatic macrophytes such as Littorella uniflora and Lobelia...... to unvegetated sediment, especially with respect to the availability of oxygen, organic carbon, and inorganic nitrogen. We hypothesize that macrophyte species create specific niches for ammonia oxidizing and nitrate-reducing bacteria in their rhizosphere, leading to plant-dependant differences in...... community composition of ammonia oxidizing bacteria between the root surface, the rhizosphere and unvegetated sediment and between plant species, however, differences in the community composition among sampling sites also suggest a strong impact of the chemical properties of the sediment....

  5. Selection of denitrifying phosphorous accumulating organisms in IFAS systems: comparison of nitrite with nitrate as an electron acceptor.

    Jabari, Pouria; Munz, Giulio; Oleszkiewicz, Jan A

    2014-08-01

    Nitrite and nitrate were compared as electron acceptors to select for denitrifying phosphorous accumulating organisms (DPAO) in two integrated fixed film activated sludge (IFAS 1 and IFAS 2) systems operated as sequencing batch reactors. The bench-scale experiment lasted one year and synthetic wastewater was used as feed. During anoxic conditions 20mgNO3(-)-NL(-1) were dosed into IFAS-1 and 20mgNO2(-)-NL(-1) were dosed into IFAS-2. Long term phosphorous and ammonia removal via nitritation were achieved in both systems and both attached and suspended biomass contributed to phosphorous and ammonia removal. DPAO showed no specific adaptation to the electron acceptor as evidenced by short term switch of feeding with nitrate or nitrite. Anoxic phosphorus uptake rate was significantly higher with nitrite than with nitrate. Results showed that DPAO activity with nitrite could be integrated into attached and suspended biomass of IFAS systems in long term operation. PMID:24873702

  6. Community structures and activity of denitrifying microbes in a forested catchment in central Japan: survey using nitrite reductase genes

    Ohte, N.; Aoki, M.; Katsuyama, C.; Suwa, Y.; Tange, T.

    2012-12-01

    To elucidate the mechanisms of denitrification processes in the forested catchment, microbial ecological approaches have been applied in an experimental watershed that has previously investigated its hydrological processes. The study catchment is located in the Chiba prefecture in central Japan under the temperate Asian monsoon climate. Potential activities of denitrification of soil samples were measured by incubation experiments under anoxic condition associated with Na15NO3 addition. Existence and variety of microbes having nitrite reductase genes were investigated by PCR amplification, cloning and sequencings of nirK and nirS fragments after DNA extraction. Contrary to our early expectation that the potential denitrification activity was higher at deeper soil horizon with consistent groundwater residence than that in the surface soil, denitrification potential was higher in shallower soil horizons than deeper soils. This suggested that the deficiency of NO3- as a respiratory substrate for denitrifier occurred in deeper soils especially in the summer. However, high denitrification activity and presence of microbes having nirK and nirS in surface soils usually under aerobic condition was explainable by the fact that the majority of denitrifying bacteria have been recognized as a facultative anaerobic bacterium. This also suggests the possibility of that denitrification occurs even in the surface soils if the wet condition is provided by rainwater during and after a storm event. Community structures of microbes having nirK were different between near surface and deeper soil horizons, and ones having nirS was different between saturated zone (under groundwater table) and unsaturated soil horizons. These imply that microbial communities with nisK are sensitive to the concentration of soil organic matters and ones with nirS is sensitive to soil moisture contents.

  7. Transitions in nirS-type Denitrifier Diversity, Community Composition, and Biogeochemical Activity along the Chesapeake Bay Estuary

    ChristopherAFrancis

    2013-08-01

    Full Text Available Chesapeake Bay, the largest estuary in North America, can be characterized as having steep and opposing gradients in salinity and dissolved inorganic nitrogen along the main axis of the Bay. In this study, the diversity of nirS gene fragments (encoding cytochrome cd1-type nitrite reductase, physical/chemical parameters, and benthic N2-fluxes were analyzed in order to determine how denitrifier communities and biogeochemical activity vary along the estuary salinity gradient. The nirS gene fragments were PCR-amplified, cloned, and sequenced from sediment cores collected at five stations. Sequence analysis of 96 to 123 nirS clones from each station revealed extensive overall diversity in this estuary, as well as distinct spatial structure in the nirS sequence distributions. Both nirS-based richness and community composition varied among stations, with the most dramatic shifts occurring between low-salinity (oligohaline and moderate-salinity (mesohaline sites. For four samples collected in April, the nirS-based richness, nitrate concentrations, and N2-fluxes all decreased in parallel along the salinity gradient from the oligohaline northernmost station to the highest salinity (polyhaline station near the mouth of the Bay. The vast majority of the 550 nirS sequences were distinct from cultivated denitrifiers, although many were closely related to environmental clones from other coastal and estuarine systems. Interestingly, 8 of the 172 OTUs identified accounted for 42% of the total nirS clones, implying the presence of a few dominant and many rare genotypes, which were distributed in a non-random manner along the salinity gradient of Chesapeake Bay. These data, comprising the largest dataset to investigate nirS clone sequence diversity from an estuarine environment, also provided information that was required for the development of nirS microarrays to investigate the interaction of microbial diversity, environmental gradients, and biogeochemical activity.

  8. Whole-Genome Transcriptional Analysis of Chemolithoautotrophic Thiosulfate Oxidation by Thiobacillus denitrificans Under Aerobic vs. Denitrifying Conditions

    Beller, H R; Letain, T E; Chakicherla, A; Kane, S R; Legler, T C; Coleman, M A

    2006-04-22

    Thiobacillus denitrificans is one of the few known obligate chemolithoautotrophic bacteria capable of energetically coupling thiosulfate oxidation to denitrification as well as aerobic respiration. As very little is known about the differential expression of genes associated with ke chemolithoautotrophic functions (such as sulfur-compound oxidation and CO2 fixation) under aerobic versus denitrifying conditions, we conducted whole-genome, cDNA microarray studies to explore this topic systematically. The microarrays identified 277 genes (approximately ten percent of the genome) as differentially expressed using Robust Multi-array Average statistical analysis and a 2-fold cutoff. Genes upregulated (ca. 6- to 150-fold) under aerobic conditions included a cluster of genes associated with iron acquisition (e.g., siderophore-related genes), a cluster of cytochrome cbb3 oxidase genes, cbbL and cbbS (encoding the large and small subunits of form I ribulose 1,5-bisphosphate carboxylase/oxygenase, or RubisCO), and multiple molecular chaperone genes. Genes upregulated (ca. 4- to 95-fold) under denitrifying conditions included nar, nir, and nor genes (associated respectively with nitrate reductase, nitrite reductase, and nitric oxide reductase, which catalyze successive steps of denitrification), cbbM (encoding form II RubisCO), and genes involved with sulfur-compound oxidation (including two physically separated but highly similar copies of sulfide:quinone oxidoreductase and of dsrC, associated with dissimilatory sulfite reductase). Among genes associated with denitrification, relative expression levels (i.e., degree of upregulation with nitrate) tended to decrease in the order nar > nir > nor > nos. Reverse transcription, quantitative PCR analysis was used to validate these trends.

  9. City model enrichment

    Smart, Philip D.; Quinn, Jonathan A.; Jones, Christopher B.

    The combination of mobile communication technology with location and orientation aware digital cameras has introduced increasing interest in the exploitation of 3D city models for applications such as augmented reality and automated image captioning. The effectiveness of such applications is, at present, severely limited by the often poor quality of semantic annotation of the 3D models. In this paper, we show how freely available sources of georeferenced Web 2.0 information can be used for automated enrichment of 3D city models. Point referenced names of prominent buildings and landmarks mined from Wikipedia articles and from the OpenStreetMaps digital map and Geonames gazetteer have been matched to the 2D ground plan geometry of a 3D city model. In order to address the ambiguities that arise in the associations between these sources and the city model, we present procedures to merge potentially related buildings and implement fuzzy matching between reference points and building polygons. An experimental evaluation demonstrates the effectiveness of the presented methods.

  10. Enrichment operations outlook

    The U. S. gaseous diffusion plants have achieved an admirable record of performance and reliability, and there is confidence it can be maintained through the rest of this century. In addition to aggressive regular maintenance, three specific programs are combined to maximize the reliability of the plants: A Deferred Maintenance Program to restore production equipment and associated auxiliary systems; a Cascade Improvement Program (CIP) to install new and improved technology such as rebuilt and more efficient compressors, piping and control valves with better aerodynamic characteristics, and new and improved barrier; and a Cascade Uprating Program (CUP) to refurbish and add capacity to electrical and process cooling systems. A three-plant steering committee has been formed to examine in detail all aspects of enrichment facilities not already covered by these three specific programs, and task forces have been assigned to investigate in depth long-term reliability in these areas. In many cases, specific testing programs have been designed to evaluate the potential life of components, and information from commercial industries is routinely used. Where deficiencies are found, capital budget items are submitted and corrective action planned. Existing experience is available to those industries concerned with new plant ventures, and the Industrial Participation Program is one example where private industry can take full advantage of this existing experience. (U.S.)

  11. Uranium enrichment and weapon proliferation

    The internationalization of the commercial enrichment industry under a unified authority is recommended. This would guarantee fuel supplies to all countries at fair prices, could arrange for equitable compensation of states with existing enrichment capabilities, and could enforce non-proliferation measures. (U.K.)

  12. Structural Monitoring of Oligosaccharides through 13C Enrichment and NMR Observation of Acetyl Groups

    YU, FEI; Prestegard, J H

    2006-01-01

    Structural characterization of biomolecules by NMR methods frequently requires the enrichment of magnetically active isotopes at particular molecular sites. Introduction is usually achieved biosynthetically through the use of bacterial cultures grown on isotopically enriched media, but for certain types of molecules—cell-surface carbohydrates of mammalian origin, for example—this is not practical. Here we explore a means of introducing 13C-enriched sites, postisolation in natural carbohydrate...

  13. Process evaluation by isotope enrichment

    An evaluation of an engineering or biological process or system can be made by treating the system with an isotopically-enriched substance which flows through the system intact or which is changed into another substance containing isotope-enrichment, and a gaseous sample thereof is analyzed at low pressure e.g. by infra-red spectroscopy to measure the intensity of a specific absorption line of the enriched isotopic species whereby the concentration of the species is calculated. The enrichment of the measured species furnishes information about the functioning of the process. In bio-medical testing, the tracer isotopic species used frequently are enriched values of CO, CO2, H2O or NH3 in the breath or derived from tissues or other specimens. (author)

  14. AEC determines uranium enrichment policy

    The Advisory Committee on Uranium Enrichment of the Atomic Energy Commission (AEC) has submitted a report to AEC chairman concerning the promotion of the introduction of advanced material, high performance centrifuges to replace conventional metallic drum centrifuges, and the development of next generation advanced centrifuges. The report also called for the postponement until around 1997 of the decision whether the development should be continued or not on atomic vapor laser isotope separation (AVLIS) and molecular laser isotope separation (MLIS) processes, as well as the virtual freezing of the construction of a chemical process demonstration plant. The report was approved by the AEC chairman in August. The uranium enrichment service market in the world will continue to be characterized by oversupply. The domestic situation of uranium enrichment supply-demand trend, progress of the expansion of Rokkasho enrichment plant, the trend in the development of gas centrifuge process and the basic philosophy of commercializing domestic uranium enrichment are reported. (K.I.)

  15. Mineralization of Diethylthiophosphoric Acid by an Enriched Consortium from Cattle Dip

    Shelton, Daniel R.

    1988-01-01

    Enrichment cultures were initiated from cattle dip solution with diethylthiophosphoric acid (DETP) as the carbon and energy source. An enriched consortium consisting of at least six distinct bacterial species was subsequently obtained. The consortium mineralized DETP to sulfate and phosphate while utilizing ethyl moieties as a carbon and energy source. These data demonstrate that DETP can be metabolized under environmentally realistic conditions.

  16. Hydrogen-enriched fuels

    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.

  17. High enrichment to low enrichment core's conversion. Accidents analysis

    This work analyzes the different accidents that may occur in the reactor's facility after the 20% high-enriched uranium core's conversion. The reactor (of 5 thermal Mw), built in the 50's and 60's, is of the 'swimming pool' type, with light water and fuel elements of the curve plates MTR type, enriched at 93.15 %. This analysis includes: a) accidents by reactivity insertion; b) accidents by coolant loss; c) analysis by flow loss and d) fission products release. (Author)

  18. High enrichment to low enrichment core's conversion. Technical securities

    This work presents the fulfillment of the technical securities subscribed by INVAP S.E. for the conversion of a high enriched uranium core. The reactor (of 5 thermal Mw), built in the 50's and 60's, is of the 'swimming pool' type, with light water and fuel elements of the curve plates MTR type, enriched at 93.15 %. These are neutronic and thermohydraulic securities. (Author)

  19. Safety of uranium enrichment plant

    With respect to safety evaluation of the gas centrifuge enrichment facility, several characteristic problems are described as follows. Criticality safety in the cascade equipments can be obtained to maintain the enrichment of UF6 below 5 %. External radiation dose equivalent rate of the 30B cylinder is low enough, the shield is not necessary. Penetration ratio of the two-stage HEPA filters for UF6 aerosol is estimated at 10-9. From the experimental investigation, vacuum tightness is not damaged by destruction of gas centrifuge rotor. Carbon steel can be used for uranium enrichment equipments under the condition below 100degC. (author)

  20. Spectral enrichments of model categories

    Dugger, D

    2005-01-01

    We prove that every stable, combinatorial model category has a natural enrichment by symmetric spectra (or more precisely, a natural equivalence class of enrichments). This in some sense generalizes the simplicial enrichments of model categories provided by the Dwyer-Kan hammock localization. As one particular application, we are able to associate to every object of a stable, combinatorial model category a "homotopy endomorphism ring spectrum". The homotopy types of these ring spectra are preserved by Quillen equivalences, and so are an invariant for stable model categories.

  1. Rectoanal Mucosal Swab Culture Is More Sensitive Than Fecal Culture and Distinguishes Escherichia coli O157:H7-Colonized Cattle and Those Transiently Shedding the Same Organism

    Rice, Daniel H.; Sheng, Haiqing Q.; Wynia, Stacey A.; Carolyn J. Hovde

    2003-01-01

    Enrichment and direct (nonenrichment) rectoanal mucosal swab (RAMS) culture techniques were developed and compared to traditional fecal culture for the detection of Escherichia coli O157:H7 in experimentally infected and naturally infected cattle. Holstein steers (n = 16) orally dosed with E. coli O157:H7 were sampled after bacterial colonization starting 15 days postinoculation. Enrichment RAMS cultures (70.31% positive) were more sensitive than enrichment fecal cultures with 10 g of feces (...

  2. Evaluation of the microbial diversity in sequencing batch reactor treating linear alkylbenzene sulfonate under denitrifying and mesophilic conditions using swine sludge as inoculum

    Iolanda Cristina Silveira Duarte

    2015-06-01

    Full Text Available The objective of this study was to evaluate the degradation of Linear Alkylbenzene Sulfonate (LAS in anaerobic sequencing batch reactor (ASBR under denitrifying conditions using swine sludge as inoculum. The reactor was operated for 104 days with synthetic substrate containing nitrate, and LAS was added later (22 mg/L. Considering the added mass of the LAS, the adsorbed mass in the sludge and discarded along with the effluent, degradation of the surfactant at the end of operation was 87%, removal of chemical oxygen demand was 86% and nitrate was 98%. The bacterial community was evaluated by cutting the bands and sequencing of polymerase chain reaction (PCR fragments and denaturing gradient gel electrophoresis (DGGE. The sequences obtained were related to the phylum Proteobacteria and the alpha-and beta-proteobacteria classes, these bacteria were probably involved in the degradation of LAS. The efficiently degraded LAS in the reactor was operated in batch sequences in denitrifying conditions.

  3. Evaluation of the microbial diversity in sequencing batch reactor treating linear alkylbenzene sulfonate under denitrifying and mesophilic conditions using swine sludge as inoculum

    Iolanda Cristina Silveira, Duarte; Lorena Lima de, Oliveira; Dagoberto Yukio, Okada; Pierre Ferreira do, Prado; Maria Bernadete Amncio, Varesche.

    2015-06-01

    Full Text Available The objective of this study was to evaluate the degradation of Linear Alkylbenzene Sulfonate (LAS) in anaerobic sequencing batch reactor (ASBR) under denitrifying conditions using swine sludge as inoculum. The reactor was operated for 104 days with synthetic substrate containing nitrate, and LAS was [...] added later (22 mg/L). Considering the added mass of the LAS, the adsorbed mass in the sludge and discarded along with the effluent, degradation of the surfactant at the end of operation was 87%, removal of chemical oxygen demand was 86% and nitrate was 98%. The bacterial community was evaluated by cutting the bands and sequencing of polymerase chain reaction (PCR) fragments and denaturing gradient gel electrophoresis (DGGE). The sequences obtained were related to the phylum Proteobacteria and the alpha-and beta-proteobacteria classes, these bacteria were probably involved in the degradation of LAS. The efficiently degraded LAS in the reactor was operated in batch sequences in denitrifying conditions.

  4. Denitrifying Alphaproteobacteria from the Arabian Sea That Express nosZ, the Gene Encoding Nitrous Oxide Reductase, in Oxic and Suboxic Waters

    Wyman, Michael; Hodgson, Sylvia; Bird, Clare

    2013-01-01

    Marine ecosystems are significant sources of the powerful greenhouse gas nitrous oxide (N2O). A by-product of nitrification and an intermediate in the denitrification pathway, N2O is formed primarily in oxygen-deficient waters and sediments. We describe the isolation of a group of alphaproteobacteria from the suboxic waters of the Arabian Sea that are phylogenetically affiliated with Labrenzia spp. and other denitrifiers. Quantitative PCR assays revealed that these organisms were very broadly...

  5. Abundance and Diversity of Bacterial Nitrifiers and Denitrifiers and Their Functional Genes in Tannery Wastewater Treatment Plants Revealed by High-Throughput Sequencing

    Wang, Zhu; Zhang, Xu-Xiang; LU, Xin; LIU, Bo; LI, YAN; Long, Chao; Li, Aimin

    2014-01-01

    Biological nitrification/denitrification is frequently used to remove nitrogen from tannery wastewater containing high concentrations of ammonia. However, information is limited about the bacterial nitrifiers and denitrifiers and their functional genes in tannery wastewater treatment plants (WWTPs) due to the low-throughput of the previously used methods. In this study, 454 pyrosequencing and Illumina high-throughput sequencing, combined with molecular methods, were used to comprehensively ch...

  6. Environmental enrichment for laboratory marmosets.

    Byron, J K; Bodri, M S

    2001-09-01

    The authors report on using two environmental enrichment devices for marmosets, and suggest the design of five other devices that may be more successful in stimulating foraging or grooming behavior than the devices tested. PMID:11910416

  7. Uranium enrichment program of NUCLEI

    The first step towards enrichment uranium in Brazil was taken by NUCLEI in 1977, in the scope of the Agreement for Cooperation in the Field of Pacific Uses of Nuclear Energy, signed with the Federal Republic of Germany in 1975. The construction of the first cascade of a demonstration plant by the Jet-Nozzle method was started in 1978 and shows, in october 1989, a physical progress of round 93%. On the results obtained from the first cascade operation - estimated for 1991 - depends NUCLEI's future, the future of its already desimbursed investments and the future availability, in the long range, of enriched uranium for the brazilian reactors. Alternatives for future enrichment programs are presented, considering that any future program for uranium enrichment necessarily passes by the first cascade commissioning and operation. (author)

  8. The enrichment industry reaches maturity

    As the nuclear power industry enters the 1980s, uranium enrichment supply can no longer be considered one of the critical problem areas of the nuclear fuel cycle. It has become an industrial and commercial activity which has reached a high degree of maturity. Three main aspects of this maturity are discussed: 1. the availability of enrichment services from several facilities with very diverse ownership; 2. the involvement of private industry, especially in Europe, and the application of normal commercial rules to enrichment contracts; 3. the ability of the enrichment industry to cope with recent setbacks in the advancement of nuclear power programmes whilst carrying out an active research and development programme that will help to ensure its future technical and economic viability. (U.K.)

  9. Enrichment and terminal differentiation of striated muscle progenitors in vitro

    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.

  10. Culturally Responsive Teaching for American Indian Learners.

    Pewewardy, Cornel D.

    Teachers in a multicultural society need to respect cultural differences, know the cultural resources their students bring to class, and be skilled at tapping into learners' cultural resources in the teaching-learning process. They must believe that all students are capable of learning, and they must implement an enriched curriculum for all…

  11. Uranium enrichment: an evolving market

    With over half of the world uranium enrichment market uncommitted to any supplier early in the next century, competition is certain to be fierce. In the meantime the outlood remains unclear, with the market dominated by a number of developments -privatisation of the United States Enrichment Corp (USEC), increasing availability of Russian and US military inventories, the deployment of advanced technology and the closure of nuclear power plants due to deregulation. (author)

  12. Uranium enrichment by displacement chromatography

    Natural Uranium contains 0.72% of fissile isotope 235U. Need for 235U enriched uranium for light water moderated reactors and other uses are well known. Uranium enrichment is a highly complex and tedious process owing to extremely small separation factor. Several processes have been tried; however, gaseous diffusion and centrifugation techniques based on mass dependent isotope effect are being exploited for producing enriched uranium. In the recent past, Isotope fractionation based on mass independent isotope effect such as LASER technique, ion exchange displacement chromatography etc. have drawn the attention of separation scientist and efforts are being made for development and industrial deployment of these technologies. Indian Nuclear Power Programme is based on use of natural uranium (PHWRs) and slightly enriched uranium (BWRs, PWRs) with emphasis on use of vast resources of thorium. Spent fuel from PHWRs, which is depleted in 235U (0.2 to 0.3%), appears to be a promising source of slightly enriched uranium (1 to 3% 235U). Harnessing this source for 235U enrichment would augment fuel supply for PHWR and light water reactors. Ion exchange displacement chromatography is a proliferation resistant novel technique for 235U enrichment. It is based on nuclear field shift - an isotope shift in orbital electrons resulting from the isotopic difference in nuclear size and shape. Due to this effect lighter isotope (235U) exhibits a stronger tendency for a higher valence i.e. UO22+ as compared to heavier isotope (238U). Preliminary laboratory study is being carried out at HWP, Manuguru for exploring feasibility of using ion exchange displacement chromatography for enriching 235U using 4-vinyl pyridine based anion exchange resin

  13. Uranium enrichment: an evolving market

    Longenecker, J.; Witzel, R. [Longenecker and Associates, Del Mar, CA (United States)

    1997-09-01

    With over half of the world uranium enrichment market uncommitted to any supplier early in the next century, competition is certain to be fierce. In the meantime the outlood remains unclear, with the market dominated by a number of developments -privatisation of the United States Enrichment Corp (USEC), increasing availability of Russian and US military inventories, the deployment of advanced technology and the closure of nuclear power plants due to deregulation. (author).

  14. Linking N2O emission from biochar-amended composting process to the abundance of denitrify (nirK and nosZ) bacteria community.

    Li, Shuqing; Song, Lina; Jin, Yaguo; Liu, Shuwei; Shen, Qirong; Zou, Jianwen

    2016-12-01

    Manure composting has been recognized as an important anthropogenic source of nitrous oxide (N2O) contributing to global warming. However, biochar effect on N2O emissions from manure composting is rarely evaluated, especially by linking it to abundance of denitrifying bacteria community. Results of this study indicated that biochar amendment significantly reduced N2O emissions from manure composting, primarily due to suppression of the nirK gene abundance of denitrifying bacteria. Pearson's correlation analysis showed a significant positive correlation between nirK abundance and N2O fluxes, while a negative correlation between nosZ density and N2O fluxes. Simultaneously, a linear correlation between nirK gene abundance minus nosZ gene abundance with N2O fluxes was also observed. In addition, a statistical model for estimating N2O emissions based on the bacterial denitrifying functional genes was developed and verified to adequately fit the observed emissions. Our results highlighted that biochar amendment would be an alternative strategy for mitigating N2O emissions during manure composting, and the information of related functional bacterial communities could be helpful for understanding the mechanism of N2O emissions. PMID:27207069

  15. Effects of carbon-to-sulfur (C/S) ratio and nitrate (N) dosage on Denitrifying Sulfur cycle-associated Enhanced Biological Phosphorus Removal (DS-EBPR)

    Yu, Mei; Lu, Hui; Wu, Di; Zhao, Qing; Meng, Fangang; Wang, Yudan; Hao, Xiaodi; Chen, Guang-Hao

    2016-03-01

    In this study, the Denitrifying Sulfur cycle-associated Enhanced Biological Phosphorous Removal (DS-EBPR) with 20 mg P/L/d of the volumetric P removal rate was successfully achieved in a Sequencing Batch Reactor (SBR). The effects of carbon-to-sulfur (C/S) mass ratio and nitrate (N) dosage were investigated through two batch tests to reveal the role of wastewater compositions in DS-EBPR performance. The optimal specific P release and uptake rates (0.4 and 2.4 mg P/g VSS/h, respectively) were achieved at C/S/P/N mass ratio of 150/200/20/20, and poly-S is supplied as a potential electron and energy storage. The nitrate dosage in a range of 10–50 mg N/L had no significant influence on P uptake rates (2.1 ~ 2.4 mg P/g VSS/h), but significantly affected the storage of inclusion poly-S, the poly-S oxidation rate was increased about 16% while dosing nitrate from 20 to 30 mg N/L. It implies that nitrate is denitrified in the P uptake phase, and excess nitrate is further consumed by poly-S. Moreover, the microbial analysis showed that the functional bacteria should mostly belong to denitrifying bacteria or Unclassified genera.

  16. Effects of carbon-to-sulfur (C/S) ratio and nitrate (N) dosage on Denitrifying Sulfur cycle-associated Enhanced Biological Phosphorus Removal (DS-EBPR).

    Yu, Mei; Lu, Hui; Wu, Di; Zhao, Qing; Meng, Fangang; Wang, Yudan; Hao, Xiaodi; Chen, Guang-Hao

    2016-01-01

    In this study, the Denitrifying Sulfur cycle-associated Enhanced Biological Phosphorous Removal (DS-EBPR) with 20 mg P/L/d of the volumetric P removal rate was successfully achieved in a Sequencing Batch Reactor (SBR). The effects of carbon-to-sulfur (C/S) mass ratio and nitrate (N) dosage were investigated through two batch tests to reveal the role of wastewater compositions in DS-EBPR performance. The optimal specific P release and uptake rates (0.4 and 2.4 mg P/g VSS/h, respectively) were achieved at C/S/P/N mass ratio of 150/200/20/20, and poly-S is supplied as a potential electron and energy storage. The nitrate dosage in a range of 10-50 mg N/L had no significant influence on P uptake rates (2.1 ~ 2.4 mg P/g VSS/h), but significantly affected the storage of inclusion poly-S, the poly-S oxidation rate was increased about 16% while dosing nitrate from 20 to 30 mg N/L. It implies that nitrate is denitrified in the P uptake phase, and excess nitrate is further consumed by poly-S. Moreover, the microbial analysis showed that the functional bacteria should mostly belong to denitrifying bacteria or Unclassified genera. PMID:26983801

  17. Effects of warming and drought on potential N2O emissions and denitrifying bacteria abundance in grasslands with different land-use.

    Keil, Daniel; Niklaus, Pascal A; von Riedmatten, Lars R; Boeddinghaus, Runa S; Dormann, Carsten F; Scherer-Lorenzen, Michael; Kandeler, Ellen; Marhan, Sven

    2015-07-01

    Increased warming in spring and prolonged summer drought may alter soil microbial denitrification. We measured potential denitrification activity and denitrifier marker gene abundances (nirK, nirS, nosZ) in grasslands soils in three geographic regions characterized by site-specific land-use indices (LUI) after warming in spring, at an intermediate sampling and after summer drought. Potential denitrification was significantly increased by warming, but did not persist over the intermediate sampling. At the intermediate sampling, the relevance of grassland land-use intensity was reflected by increased potential N2O production at sites with higher LUI. Abundances of total bacteria did not respond to experimental warming or drought treatments, displaying resilience to minor and short-term effects of climate change. In contrast, nirS- and nirK-type denitrifiers were more influenced by drought in combination with LUI and pH, while the nosZ abundance responded to the summer drought manipulation. Land-use was a strong driver for potential denitrification as grasslands with higher LUI also had greater potentials for N2O emissions. We conclude that both warming and drought affected the denitrifying communities and the potential denitrification in grassland soils. However, these effects are overruled by regional and site-specific differences in soil chemical and physical properties which are also related to grassland land-use intensity. PMID:26092950

  18. Assessing the activity and diversity of fumarate-fed denitrifying bacteria by performing field single-well push-pull tests.

    Kim, Jin-Hoon; Ha, Chul-Yoon; Oa, Seong-Wook; Lee, Jin-Woo; Park, Sun-Hwa; Kwon, Soo-Youl; Kim, Sungpyo; Kim, Young

    2011-01-01

    In situ biological denitrification has been proposed as an important metabolic activity in the remediation of nitrate-contaminated groundwater. In this study, the effects of fumarate, an electron donor for biological denitrification, on the in situ denitrifying activity were determined by using three types of single-well push-pull tests; transport, biostimulation and activity tests. During the tests, changes in microbial community composition were also investigated using denaturing gradient gel electrophoresis (DGGE) of 16S rRNA genes. Transport test demonstrated that non-reactive tracer and biologically reactive solutes behaved similarly. A biostimulation test was conducted to stimulate the denitrifying activities of native microorganisms, which were monitored by detecting the simultaneous production of CO(2) and drastic degradations of both nitrate and fumarate after the injection of fumarate as an electron donor and/or carbon source, with nitrate as an electron acceptor. A phylogenetic analysis suggested that the taxonomic affiliation of the dominant species before biostimulation was γ-Proteobacteria, including Acinetobacter species and Pseudomonas fluorescens, while the dominant species after biostimulation were affiliated with β-Proteobacteria, cytophaga-Flavobacterium-Bacteroides and high G+C gram-positive bacteria. These results suggest that the analyses of groundwater samples using a combination of single well push pull tests with DGGE can be applied to investigate the activity, diversity and composition shift of denitrifying bacteria in a nitrate-contaminated aquifer. PMID:21104493

  19. Abundance and Diversity of Bacterial Nitrifiers and Denitrifiers and Their Functional Genes in Tannery Wastewater Treatment Plants Revealed by High-Throughput Sequencing

    Wang, Zhu; Zhang, Xu-Xiang; Lu, Xin; Liu, Bo; Li, Yan; Long, Chao; Li, Aimin

    2014-01-01

    Biological nitrification/denitrification is frequently used to remove nitrogen from tannery wastewater containing high concentrations of ammonia. However, information is limited about the bacterial nitrifiers and denitrifiers and their functional genes in tannery wastewater treatment plants (WWTPs) due to the low-throughput of the previously used methods. In this study, 454 pyrosequencing and Illumina high-throughput sequencing, combined with molecular methods, were used to comprehensively characterize structures and functions of nitrification and denitrification bacterial communities in aerobic and anaerobic sludge of two full-scale tannery WWTPs. Pyrosequencing of 16S rRNA genes showed that Proteobacteria and Synergistetes dominated in the aerobic and anaerobic sludge, respectively. Ammonia-oxidizing bacteria (AOB) amoA gene cloning revealed that Nitrosomonas europaea dominated the ammonia-oxidizing community in the WWTPs. Metagenomic analysis showed that the denitrifiers mainly included the genera of Thauera, Paracoccus, Hyphomicrobium, Comamonas and Azoarcus, which may greatly contribute to the nitrogen removal in the two WWTPs. It is interesting that AOB and ammonia-oxidizing archaea had low abundance although both WWTPs demonstrated high ammonium removal efficiency. Good correlation between the qPCR and metagenomic analysis is observed for the quantification of functional genes amoA, nirK, nirS and nosZ, indicating that the metagenomic approach may be a promising method used to comprehensively investigate the abundance of functional genes of nitrifiers and denitrifiers in the environment. PMID:25420093

  20. Nitrite production in a partial denitrifying upflow sludge bed (USB) reactor equipped with gas automatic circulation (GAC).

    Cao, Shenbin; Li, Baikun; Du, Rui; Ren, Nanqi; Peng, Yongzhen

    2016-03-01

    Nitrite production in a partial denitrifying (NO3(-)-N→NO2(-)-N) upflow sludge bed (USB) reactor equipped with gas automatic circulation (GAC) was investigated at ambient temperature (28.8-14.1 °C). The nitrite production rate (NPR) increased with the nitrate loading rate (NLR). Average NPR of 6.63 kgN m(-3) d(-1) was obtained at 28.0 °C with the organic loading rate (OLR) and NLR of 25.38 KgCOD∙m(-3)∙d(-1) and 10.82 kgN m(-3) d(-1), respectively. However, serious sludge floatation was observed when the NLR increased to 13.18 kgN m(-3) d(-1), which might be attributed to sludge bulking at high NLR. The USB reactor recovered rapidly when seeded with the sludge discharged before the deteriorated period, and a stable NPR of ∼4.35 kgN m(-3) d(-1) was achieved at 14.1-15.7 °C in the following 100-day operation, during which the maximum nitrate-to-nitrite transformation ratio (NTR) of 81.4% was achieved at the GAC rate of 1.08 L h(-1). The application of GAC in the partial denitrifying USB reactor enhanced mass transfer, which effectively avoided the channel and dead space, and improved the nitrate transform to nitrite. Moreover, it was found that the GAC system played an important role in promoting the stability of the USB reactor by preventing the sludge floatation. The Illumina high-throughput sequencing analysis revealed that the genus of Thauera was dominate in the USB reactor (67.2-50.2%), which may be responsible for the high nitrite accumulation. Results in this study have an important application in treating nitrate wastewater with an economic and efficient way by combining with ANAMMOX process. PMID:26760483

  1. The activity and community structure of total bacteria and denitrifying bacteria across soil depths and biological gradients in estuary ecosystem.

    Lee, Seung-Hoon; Kang, Hojeong

    2016-02-01

    The distribution of soil microorganisms often shows variations along soil depth, and even in the same soil layer, each microbial group has a specific niche. In particular, the estuary soil is intermittently flooded, and the characteristics of the surface soil layer are different from those of other terrestrial soils. We investigated the microbial community structure and activity across soil depths and biological gradients composed of invasive and native plants in the shallow surface layer of an estuary ecosystem by using molecular approaches. Our results showed that the total and denitrifying bacterial community structures of the estuarine wetland soil differed according to the short depth gradient. In growing season, gene copy number of 16S rRNA were 1.52(±0.23) × 10(11), 1.10(±0.06) × 10(11), and 4.33(±0.16) × 10(10) g(-1) soil; nirS were 5.41(±1.25) × 10(8), 4.93(±0.94) × 10(8), and 2.61(±0.28) × 10(8) g(-1) soil; and nirK were 9.67(±2.37) × 10(6), 3.42(±0.55) × 10(6), and 2.12(±0.19) × 10(6) g(-1) soil in 0 cm, 5 cm, and 10 cm depth layer, respectively. The depth-based difference was distinct in the vegetated sample and in the growing season, evidencing the important role of plants in structuring the microbial community. In comparison with other studies, we observed differences in the microbial community and functions even across very short depth gradients. In conclusion, our results suggested that (i) in the estuary ecosystem, the denitrifying bacterial community could maintain its abundance and function within shallow surface soil layers through facultative anaerobiosis, while the total bacterial community would be both quantitatively and qualitatively affected by the soil depth, (ii) the nirS gene community, rather than the nirK one, should be the first candidate used as an indicator of the microbial denitrification process in the estuary system, and (iii) as the microbial community is distributed and plays a certain niche role according to biogeochemical factors, the study of the microbial community even in surface soil should be performed in detail by considering the soil depth. PMID:26526456

  2. Comparison of enrichment measurement techniques for gas centrifuge enrichment plants

    A comparison of enrichment techniques has been carried out using a small bank of gas centrifuge enrichment units to simulate conditions in a commercial plant. The detectors compared included an in-line version of the gas phase enrichment monitor and a prototype Go-NoGo monitor. In these comparisons it was found that the gas phase enrichment monitor had an ultimate precision of about 0.5% for all measurements of enrichment between natural and 20%. The Go-NoGo instrument developed in the present programme was based on previous studies, and took advantage of the experience gained in other support programmes. Two different methods of correcting for wall deposits were investigated. On the basis of these studies a versatile Go-NoGo instrument has been designed for usage on pipes varying in diameter from 30 to 120 mm. Further development of this instrument will be carried out using a UF6 test loop facility which is currently under construction. (author)

  3. Energetic optimization of algal lipid production in bubble columns: Part II: Evaluation of CO2 enrichment

    This study evaluated growth and lipid productivity of Nannochloropsis salina under sparging with carbon dioxide-enriched air. Carbon dioxide enrichments ranging from 0.5 to 9.5% and gas-to-culture volume ratios ranging from 0.02 to 1.0 min−1 were tested in 900 mL bubble column batch reactors. An energy-based approach is proposed to optimize CO2 enrichment. Cultures sparged with CO2-enrichments of 0.5–4% grew at nearly double the rate of those sparged with ambient air, accumulating 64% or more lipids. Based on energy efficiency, CO2-enrichment of 0.5% was found to be optimal while CO2-enrichments of 6.5% and greater were observed to cause inhibition. At this optimal CO2-enrichment of 0.5%, no significant changes were noted in the lipid content over the range of gas-to-culture volume ratios tested. These studies indicated that optimal energy recovery from N. salina can be achieved with CO2-enrichment of 0.5% sparged at a gas-to-culture volume ratio 0.18 min−1. Under this optimal condition, biomass growth rate was 0.161 g L−1 d−1, and the lipid content was 67.8%, yielding total lipid production of 0.771 g L−1 over 10 days at a net energy yield of 25 W m−3. -- Highlights: ► Evaluated algal productivity under sparging with CO2-enriched air. ► Proposed net energy to optimize algal productivity. ► Optimized CO2-enrichment and gas flow to maximize net energy. ► Net energy of 25 W/m3 found under optimal conditions.

  4. Partial Nitrification and Denitrifying Phosphorus Removal in a Pilot-Scale ABR/MBR Combined Process.

    Wu, Peng; Xu, Lezhong; Wang, Jianfang; Huang, Zhenxing; Zhang, Jiachao; Shen, Yaoliang

    2015-11-01

    A pilot-scale combined process consisting of an anaerobic baffled reactor (ABR) and an aerobic membrane bioreactor (MBR) for the purpose of achieving easy management, low energy demands, and high efficiencies on nutrient removal from municipal wastewater was investigated. The process operated at room temperature with hydraulic retention time (HRT) of 7.5 h, recycle ratio 1 of 200%, recycle ratio 2 of 100%, and dissolved oxygen (DO) of 1 mg/L and achieved good effluent quality with chemical oxygen demand (COD) of 25 mg/L, NH4 (+)-N of 4 mg/L, total nitrogen (TN) of 11 mg/L, and total phosphorus (TP) of 0.7 mg/L. The MBR achieved partial nitrification, and NO2 (-)-N has been accumulated (4 mg/L). Efficient short-cut denitrification was occurred in the ABR with a TN removal efficiency of 51%, while the role of denitrification and phosphorus removal removed partial TN (14%). Furthermore, nitrogen was further removed (11%) by simultaneous nitrification and denitrification in the MBR. In addition, phosphorus accumulating organisms in the MBR sufficiently uptake phosphorus; thus, effluent TP further reduced with a TP removal efficiency of 84%. Analysis of fluorescence in situ hybridization (FISH) showed that ammonia oxidizing bacteria (AOB) and phosphorus accumulating organisms (PAOs) were enriched in the process. In addition, the accumulation of NO2 (-)-N was contributed to the inhibition on the activities of the NOB rather than its elimination. PMID:26411352

  5. [Influences of long-term application of organic and inorganic fertilizers on the composition and abundance of nirS-type denitrifiers in black soil].

    Yin, Chang; Fan, Fen-Liang; Li, Zhao-Jun; Song, A-Lin; Zhu, Ping; Peng, Chang; Liang, Yong-Chao

    2012-11-01

    The objectives of this study were to explore the effects of long-term organic and inorganic fertilizations on the composition and abundance of nirS-type denitrifiers in black soil. Soil samples were collected from 4 treatments (i. e. no fertilizer treatment, CK; organic manure treatment, OM; chemical fertilizer treatment (NPK) and combination of organic and chemical fertilizers treatment (MNPK)) in Gongzhuling Long-term Fertilization Experiment Station. Composition and abundance of nirS-type denitrifiers were analyzed with terminal restriction fragment length polymorphism (T-RFLP) and real-time quantitative PCR (Q-PCR), respectively. Denitrification enzyme activity (DEA) and soil properties were also measured. Application of organic fertilizers (OM and MNPK) significantly increased the DEAs of black soil, with the DEAs in OM and MNPK being 5.92 and 6.03 times higher than that in CK treatment, respectively, whereas there was no significant difference between NPK and CK. OM and MNPK treatments increased the abundances of nirS-type denitrifiers by 2.73 and 3.83 times relative to that of CK treatment, respectively. The abundance of nirS-type denitrifiers in NPK treatment was not significantly different from that of CK. The T-RFLP analysis of nirS genes showed significant differences in community composition between organic and inorganic treatments, with the emergence of a 79 bp T-RF, a significant decrease in relative abundance of the 84 bp T-RF and a loss of the 99 bp T-RF in all organic treatments. Phylogenetic analysis indicated that the airS-type denitrifiers in the black soil were mainly composed of alpha, beta and gamma-Proteobacteria. The 79 bp-type denitrifiers inhabiting exclusively in organic treatments (OM and MNPK) were affiliated to Pseudomonadaceae in gamma-Proteobacteria and Burkholderiales in beta-Proteobacteria. The 84 bp-types were related to Burkholderiales and Rhodocyclales. Correlation analysis indicated that pH, concentrations of total nitrogen (TN), total phosphorus (TP), total organic carbon (TOC), nitrate (NO3(-) -N) and ammonia (NH4(+) -N) were significantly related to abundances of nirS-denitrifers (r = 0.724-0.922, P < 0.05) and the DEA (r = 0.453-0.938, P < 0.01). In addition, the DEAs were linearly and positively correlated with the abundances of nirS-type denitrifers (r = 0.85, P < 0.01). Redundancy analysis showed that except moisture, pH and concentrations of TP, TP, TOC, NH4(+) -N and NO3(-) -N were significantly correlated with the community structure of nirS-type denirifiers (r = 0.440-0.862, P < 0.01). Furthermore, the DEAs were significantly correlated with the compositions of nirS-denirifiers (r = 0.863, P < 0.01). In conclusion, the airS-type denitrifiers in the black soil are more responsive to the organic treatments than to the inorganic treatments in terms of community composition and abundance, both of which are correlated with the changes of DEAs. PMID:23323433

  6. Semantic enrichment for medical ontologies.

    Lee, Yugyung; Geller, James

    2006-04-01

    The Unified Medical Language System (UMLS) contains two separate but interconnected knowledge structures, the Semantic Network (upper level) and the Metathesaurus (lower level). In this paper, we have attempted to work out better how the use of such a two-level structure in the medical field has led to notable advances in terminologies and ontologies. However, most ontologies and terminologies do not have such a two-level structure. Therefore, we present a method, called semantic enrichment, which generates a two-level ontology from a given one-level terminology and an auxiliary two-level ontology. During semantic enrichment, concepts of the one-level terminology are assigned to semantic types, which are the building blocks of the upper level of the auxiliary two-level ontology. The result of this process is the desired new two-level ontology. We discuss semantic enrichment of two example terminologies and how we approach the implementation of semantic enrichment in the medical domain. This implementation performs a major part of the semantic enrichment process with the medical terminologies, with difficult cases left to a human expert. PMID:16185937

  7. Uranium enrichment: technology, economics, capacity

    Large-scale enrichment of uranium has now been carried out for 40 years. While the gaseous diffusion process was the original choice of several countries and continues today to provide the major component of the world production of separative work, the last two decades have witnessed the development of a number of alternative processes for enrichment. These processes, which are being studied and deployed around the world, offer a wide range of technical and economic characteristics which will be useful in assuring adequate capacity to meet projected reactor fuel market needs through the rest of this century at competitive prices. With present uncertainties in future enriched uranium needs, it is apparent that flexibility in the deployment and operation of any enrichment process will be one of the prime considerations for the future. More economical production of separative work not only can have a beneficial impact on reactor fuel costs, but also tends to conserve natural uranium resources. This paper reviews the world scene in the enrichment component of the fuel cycle, including existing or planned commercial-scale facilities and announced R and D efforts on various processes

  8. Gaseous diffusion -- the enrichment workhorse

    Construction of the first large-scale gaseous diffusion facility was started as part of the Manhattan Project in Oak Ridge, Tennessee, in 1943. This facility, code named ''K-25,'' began operation in January 1945 and was fully on stream by September 1945. Four additional process buildings were later added in Oak Ridge as the demand for enriched uranium escalated. New gaseous diffusion plants were constructed at Paducah, Kentucky, and Portsmouth, Ohio, during this period. The three gaseous diffusion plants were the ''workhorses'' which provided the entire enriched uranium demand for the United States during the 1950s and 1960s. As the demand for enriched uranium for military purposes decreased during the early 1960s, power to the diffusion plants was curtailed to reduce production. During the 1960s, as plans for the nuclear power industry were formulated, the role of the diffusion plants gradually changed from providing highly-enriched uranium for the military to providing low-enriched uranium for power reactors

  9. Why is Brazil enriching uranium?

    In Brazil construction began in 1971 on Angra 1, a 626 MW Westinghouse pressurized water reactor (PWR). It was completed in 1984. Later, Angra 2 (a Kraftwerk Union PWR) achieved commercial operation in 2000. Brazil is considering the construction of seven nuclear power plants over the next 15 years. In preparation for this nuclear industry expansion, Brazil is building a uranium enrichment facility to provide nuclear fuel for Angra 1 and 2 starting in 2010 at Resende in the state of Rio de Janeiro, and collated with nuclear fuel fabrication facilities. This paper investigates whether the Resende Enrichment Facility will be able to provide uranium enrichment services at a cost lower than the international market price. We find that while Brazil is unlikely to be internationally competitive in the enrichment market, the Resende Enrichment Facility completes the front end of Brazil's nuclear fuel cycle. This assures uninterrupted nuclear fuel to its currently operating light water reactors, while providing the option of expanding capacity, lowering cost, and competing in the international nuclear fuel market after 2020

  10. Why is Brazil enriching uranium?

    Cabrera-Palmer, Belkis [Center for International Security and Cooperation (CISAC), Stanford University, Stanford, CA 94305-6165 (United States); Rothwell, Geoffrey [Department of Economics, Stanford University, Stanford, CA 94305-6072 (United States)

    2008-07-15

    In Brazil construction began in 1971 on Angra 1, a 626 MW Westinghouse pressurized water reactor (PWR). It was completed in 1984. Later, Angra 2 (a Kraftwerk Union PWR) achieved commercial operation in 2000. Brazil is considering the construction of seven nuclear power plants over the next 15 years. In preparation for this nuclear industry expansion, Brazil is building a uranium enrichment facility to provide nuclear fuel for Angra 1 and 2 starting in 2010 at Resende in the state of Rio de Janeiro, and collated with nuclear fuel fabrication facilities. This paper investigates whether the Resende Enrichment Facility will be able to provide uranium enrichment services at a cost lower than the international market price. We find that while Brazil is unlikely to be internationally competitive in the enrichment market, the Resende Enrichment Facility completes the front end of Brazil's nuclear fuel cycle. This assures uninterrupted nuclear fuel to its currently operating light water reactors, while providing the option of expanding capacity, lowering cost, and competing in the international nuclear fuel market after 2020. (author)

  11. A laboratory investigation of interactions between denitrifying anaerobic methane oxidation (DAMO) and anammox processes in anoxic environments

    Hu, Shihu; Zeng, Raymond J.; Haroon, Mohamed F.; Keller, Jurg; Lant, Paul A.; Tyson, Gene W.; Yuan, Zhiguo

    2015-03-01

    This study investigates interactions between recently identified denitrifying anaerobic methane oxidation (DAMO) and anaerobic ammonium oxidation (anammox) processes in controlled anoxic laboratory reactors. Two reactors were seeded with the same inocula containing DAMO organisms Candidatus Methanoperedens nitroreducens and Candidatus Methylomirabilis oxyfera, and anammox organism Candidatus Kuenenia stuttgartiensis. Both were fed with ammonium and methane, but one was also fed with nitrate and the other with nitrite, providing anoxic environments with different electron acceptors. After steady state reached in several months, the DAMO process became solely/primarily responsible for nitrate reduction while the anammox process became solely responsible for nitrite reduction in both reactors. 16S rRNA gene amplicon sequencing showed that the nitrate-driven DAMO organism M. nitroreducens dominated both the nitrate-fed (~70%) and the nitrite-fed (~26%) reactors, while the nitrite-driven DAMO organism M. oxyfera disappeared in both communities. The elimination of M. oxyfera from both reactors was likely the results of this organism being outcompeted by anammox bacteria for nitrite. K. stuttgartiensis was detected at relatively low levels (1-3%) in both reactors.

  12. Biodegradability of some nitrogenous heterocyclic compounds and co-degradation with phenol by denitrifiers in anoxic sludge reactor.

    Wang, Zixing; Xu, Xiaochen; Yang, Fenglin; Tan, Zhongxia; Chen, Jie

    2015-01-01

    Phenol and nitrogenous heterocyclic compounds (NHCs) are typical organic pollutants in coal gasification wastewater which are difficult to deal with. Unlike phenol, the stable molecular structure of NHCs make them nearly impossible to degrade under aerobic or anaerobic condition. In this paper, biodegradation of phenol and NHCs as carbon sources for denitrification was studied in a laboratory-scale anoxic reactor. Denitrifiers could degrade 490 mg/L phenol and 321.5 mg/L NO3(-)-N within 12 hours with removal efficiencies of 99.8% and 99.6%, respectively. The inhibition of pyridine on the microbes could be reduced by adding phenol into influent and the experimental results showed that pyridine could be degraded as the sole carbon source with the maximum organic loading rate of 4.38 mg/(g MLSSh) (MLSS: mixed liquor suspended solids). When phenol was included as a growth substrate, the degradation performance of quinoline and pyrrole was improved due to co-degradation, and removal rate of NHCs increased according with increment of phenol in influent. PMID:26204065

  13. Isolation and Characterization of Novel Denitrifying Bacterium Geobacillus sp. SG-01 Strain from Wood Chips Composted with Swine Manure.

    Yang, Seung-Hak; Cho, Jin-Kook; Lee, Soon-Youl; Abanto, Oliver D; Kim, Soo-Ki; Ghosh, Chiranjit; Lim, Joung-Soo; Hwang, Seong-Gu

    2013-11-01

    Nitrate contamination in ground and surface water is an increasingly serious environmental problem and only a few bacterial strains have been identified that have the ability to remove nitrogen pollutants from wastewater under thermophilic conditions. We therefore isolated thermophilic facultative bacterial strains from wood chips that had been composted with swine manure under aerated high temperature conditions so as to identify strains with denitrifying ability. Nine different colonies were screened and 3 long rod-shaped bacterial strains designated as SG-01, SG-02, and SG-03 were selected. The strain SG-01 could be differentiated from SG-02 and SG-03 on the basis of the method that it used for sugar utilization. The 16S rRNA genes of this strain also had high sequence similarity with Geobacillus thermodenitrificans 465(T) (99.6%). The optimal growth temperatures (55C), pH values (pH 7.0), and NaCl concentrations (1%) required for the growth of strain SG-01 were established. This strain reduced 1.18 mM nitrate and 1.45 mM nitrite in LB broth after 48 h of incubation. These results suggest that the G. thermodenitrificans SG-01 strain may be useful in the removal of nitrates and nitrites from wastewater generated as a result of livestock farming. PMID:25049754

  14. Functional bacteria and process metabolism of the Denitrifying Sulfur conversion-associated Enhanced Biological Phosphorus Removal (DS-EBPR) system: An investigation by operating the system from deterioration to restoration.

    Guo, Gang; Wu, Di; Hao, Tianwei; Mackey, Hamish Robert; Wei, Li; Wang, Haiguang; Chen, Guanghao

    2016-05-15

    A sulfur conversion-associated Enhanced Biological Phosphorus (P) Removal (EBPR) system is being developed to cater for the increasing needs to treat saline/brackish wastewater resulting from seawater intrusion into groundwater and sewers and frequent use of sulfate coagulants during drinking water treatment, as well as to meet the demand for eutrophication control in warm climate regions. However, the major functional bacteria and metabolism in this emerging biological nutrient removal system are still poorly understood. This study was thus designed to explore the functional microbes and metabolism in this new EBPR system by manipulating the deterioration, failure and restoration of a lab-scale system. This was achieved by changing the mixed liquor suspended solids (MLSS) concentration to monitor and evaluate the relationships among sulfur conversion (including sulfate reduction and sulfate production), P removal, variation in microbial community structures, and stoichiometric parameters. The results show that the stable Denitrifying Sulfur conversion-associated EBPR (DS-EBPR) system was enriched by sulfate-reducing bacteria (SRB) and sulfide-oxidizing bacteria (SOB). These bacteria synergistically participated in this new EBPR process, thereby inducing an appropriate level of sulfur conversion crucial for achieving a stable DS-EBPR performance, i.e. maintaining sulfur conversion intensity at 15-40 mg S/L, corresponding to an optimal sludge concentration of 6.5 g/L. This range of sulfur conversion favors microbial community competition and various energy flows from internal polymers (i.e. polysulfide or elemental sulfur (poly-S(2-)/S(0)) and poly-β-hydroxyalkanoates (PHA)) for P removal. If this range was exceeded, the system might deteriorate or even fail due to enrichment of glycogen-accumulating organisms (GAOs). Four methods of restoring the failed system were investigated: increasing the sludge concentration, lowering the salinity or doubling the COD loading, non of which restored SRB and SOB activities for DS-EBPR; only the final novel approach of adding 25 ± 5 mg S/L of external sulfide into the reactor at the beginning of the anoxic phase could efficiently restore the DS-EBPR system from failure. The present study represents a step towards understanding the DS-EBPR metabolism and provides an effective remedial measure for recovering a deteriorating or failed DS-EBPR system. PMID:27010789

  15. An Ignored Perspective in Cross-cultural Teaching-the Aestheticization of Chinese Ethic Culture

    Peng Shuixiang

    2014-01-01

    Wisdom regained through perspective has become a significant mission of cross-cultural teaching and communication. It’s urgent for Chinese aesthetic conception to merge into Cross-cultural Teaching in the trend of cultural globalization, in which the aestheticization of Chinese ethic culture is often ignored. This study is to rediscover new light for today’s humanity in the traditional notions of wisdom from Chinese ethic culture and to enrich people transculturally. It is herein argued that ...

  16. Enrichment of light hydrocarbon mixture

    Yang; Dali (Los Alamos, NM); Devlin, David (Santa Fe, NM); Barbero, Robert S. (Santa Cruz, NM); Carrera, Martin E. (Naperville, IL); Colling, Craig W. (Warrenville, IL)

    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.

  17. Enrichment of light hydrocarbon mixture

    Yang, Dali; Devlin, David; Barbero, Robert S.; Carrera, Martin E.; Colling, Craig W.

    2011-11-29

    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.

  18. Advances in uranium enrichment processes

    Advances in gas centrifuges and development of the atomic vapour laser isotope separation process promise substantial reductions in the cost of enriched uranium. The resulting reduction in LWR fuel costs could seriously erode the economic advantage of CANDU, and in combination with LWR design improvements, shortened construction times and increased operational reliability could allow the LWR to overtake CANDU. CANDU's traditional advantages of neutron economy and high reliability may no longer be sufficient - this is the challenge. The responses include: combining neutron economy and dollar economy by optimizing CANDU for slightly enriched uranium fuel; developing cost-reducing improvements in design, manufacture and construction; and reducing the cost of heavy water. Technology is a renewable resource which must be continually applied to a product for it to remain competitive in the decades to come. Such innovation is a prerequisite to Canada increasing her share of the international market for nuclear power stations. The higher burn-up achievable with enriched fuel in CANDU can reduce the fuel cycle costs by 20 to 40 percent for a likely range of costs for yellowcake and separative work. Alternatively, some of the benefits of a higher fissile content can take the form of a cheaper reactor core containing fewer fuel channels and less heavy water, and needing only a single fuelling machine. An opportunity that is linked to this need to introduce an enriched uranium fuel cycle into CANDU is to build an enrichment business in Canada. This could offer greater value added to our uranium exports, security of supply for enriched CANDUs, technological growth in Canada and new employment opportunities. AECL has a study in progress to define this opportunity

  19. Advances in uranium enrichment processes

    Advances in gas centrifuges and development of the atomic vapour laser isotope separation process promise substantial reductions in the cost of enriched uranium. The resulting reduction in LWR fuel costs could seriously erode the economic advantage of CANDU, and in combination with LWR design improvement, shortened construction times and increased operational reliability could allow the LWR to overtake CANDU. CANDU's traditional advantages of neutron economy and high reliability may no longer be sufficient - this is the challenge. The responses include: combining neutron economy and dollar economy by optimizing CANDU for slightly enriched uranium fuel; developing cost-reducing improvements in design, manufacture and construction; and reducing the cost of heavy water. Technology is a renewable resource which must be continually applied to a product for it to remain competitive in the decades to come. Such innovation is a prerequisite to Canada increasing her share of the international market for nuclear power stations. The higher burn-up achievable with enriched fuel in CANDU can reduce the fuel cycle costs by 20 to 40 percent for a likely range of costs for yellowcake and separative work. Alternatively, some of the benefits of a higher fissile content can take the form of a cheaper reactor core containing fewer fuel channels and less heavy water, and needing only a single fuelling machine. An opportunity that is linked to this need to introduce an enriched uranium fuel cycle into CANDU is to build an enrichment business in Canada. This could offer greater value added to our uranium exports, security of supply for enriched CANDUs, technological growth in Canada and new employment opportunities. AECL has a study in progress to define this opportunity

  20. Uranium enriched granites in Sweden

    Granites with uranium contents higher than normal occur in a variety of geological settings in the Swedish Precambrian, and represent a variety of granite types and ages. They may have been generated by (1) the anatexis of continental crust (2) processes occurring at a much greater depth. They commonly show enrichement in F, Sn, W and/or Mo. Only in one case is an important uranium mineralization thought to be directly related to a uranium-enriched granite, while the majority of epigenetic uranium mineralizations with economic potential are related to hydrothermal processes in areas where the bedrock is regionally uranium-enhanced. (Authors)

  1. A new uranium enrichment facility

    The Tricastin society of uranium enrichment, Areva subsidiary began to build its new facility of uranium enrichment, George Bess 2. Based on a low energy consumer technology, this facility is devoted to replace the actual facility in 2012. The complex will allow to have available 7.5 millions of separation work units. The modular conception of the facility will allow a progressive commissioning and a continuity in the fuel production. The definitive cessation of EURODIF facility is planned for 2012. (N.C.)

  2. Uranium enrichment. 1980 annual report

    This report contains data and related information on the production of enriched uranium at the gaseous diffusion plants and an update on the construction and project control center for the gas centrifuge plant. Power usage at the gaseous diffusion plants is illustrated. The report contains several glossy color pictures of the plants and processes described. In addition to gaseous diffusion and the centrifuge process, three advanced isotope separation process are now being developed. The business operation of the enrichment plants is described; charts on revenue, balance sheets, and income statements are included

  3. Mixing Intensity Effects of Attached Growth on Enriched Anammox Cultures

    Pongsak Noophan

    2015-01-01

    Full Text Available Anaerobic ammonium oxidation (anammox is a promising new technology for the treatment of wastewater with high ammonium and low carbon concentrations. Earlier work suggests that optimal processing would be realized within a sequencing batch reactor (SBR. However, the relatively slow growth of anammox bacteria inhibits the rates of nitrogen removal and biomass yielding. Improved anammox performance has been demonstrated when the bacteria are in granular form or attached to a growth medium. Little has been reported concerning the effect of mixing rate on nitrogen (N removal with attached anammox bacteria. This work subjected anammox bacteria attached to polystyrene sponge in SBR to various intensities of impeller mixing and studied the effect on NH4+ and NO2- removal. Nitrogen processing was virtually the same with velocity gradient values between 13.5 and 222 s-1. More vigorous mixing at 407 and 666 s-1 values significantly inhibited N removal, likely due to detachment of bacteria from the growth medium. Following the poor N removal at the two higher mixing intensities, agitation was reduced to 24.8 s-1 velocity gradient value. Recovery of N removal rates required 2-3 weeks, the slow time attributed to slow reattachment to the growth medium. Denaturing gradient gel electrophoresis (DGGE analysis identified the prominent anammox species in the experimental study as Candidatus Brocadia anammoxidans and Candidatus Kuenenia stuttgartiensis.

  4. Creating a Culturally Enriched Curriculum for Grades K-6.

    Block, Cathy Collins; Zinke, Jo Ann

    This book provides a multicultural curriculum that can be used alone or as a supplemental program for language arts and social studies instruction in kindergarten through grade 6. It uses the pluralistic values of the students' heritages to develop their social and problem-solving abilities. Each of the 30 units contains quotations that exemplify…

  5. Mixing Intensity Effects of Attached Growth on Enriched Anammox Cultures

    Pongsak Noophan; Tippawan Boonyawat; Saengdao Saengsuwan; Siriporn Larpkiattaworn

    2015-01-01

    Anaerobic ammonium oxidation (anammox) is a promising new technology for the treatment of wastewater with high ammonium and low carbon concentrations. Earlier work suggests that optimal processing would be realized within a sequencing batch reactor (SBR). However, the relatively slow growth of anammox bacteria inhibits the rates of nitrogen removal and biomass yielding. Improved anammox performance has been demonstrated when the bacteria are in granular form or attached to a growth medium. Li...

  6. Two-Stage Mineralization of Phenanthrene by Estuarine Enrichment Cultures

    Guerin, William F.; Jones, Galen E.

    1988-01-01

    The polycyclic aromatic hydrocarbon phenanthrene was mineralized in two stages by soil, estuarine water, and sediment microbial populations. At high concentrations, phenanthrene was degraded, with the concomitant production of biomass and accumulation of Folin-Ciocalteau-reactive aromatic intermediates. Subsequent consumption of these intermediates resulted in a secondary increase in biomass. Analysis of intermediates by high-performance liquid chromatography, thin-layer chromatography, and U...

  7. Isotopomeric characterization of nitrous oxide produced by reaction of enzymes extracted from nitrifying and denitrifying bacteria

    Yamazaki, T.; Hozuki, T.; Arai, K.; Toyoda, S.; Koba, K.; Fujiwara, T.; Yoshida, N.

    2014-05-01

    Nitrous oxide (N2O) is a potent greenhouse gas and produced in denitrification and nitrification by various microorganisms. Site preference (SP) of 15N in N2O, which is defined as the difference in the natural abundance of isotopomers 14N15NO and 15N14NO relative to 14N14NO, has been reported to be a useful tool to quantitatively distinguish N2O production pathways. To determine representative SP values for each microbial process, we firstly measured SP of N2O produced in the enzyme reaction of hydroxylamine oxidoreductase (HAO) purified from two species of ammonia oxidizing bacteria (AOB), Nitrosomonas europaea and Nitrosococcus oceani, and that of nitric oxide reductase (NOR) from Paracoccus denitrificans. The SP value for NOR reaction (-5.9 ± 2.1‰) showed nearly the same value as that reported for N2O produced by P. denitrificans in pure culture. In contrast, SP value for HAO reaction (36.3 ± 2.3‰) was a little higher than the values reported for N2O produced by AOB in aerobic pure culture. Using the SP values obtained by HAO and NOR reactions, we calculated relative contribution of the nitrite (NO2-) reduction (which is followed by NO reduction) to N2O production by N. oceani incubated under different O2 availability. Our calculations revealed that previous in vivo studies might have underestimated the SP value for the NH2OH oxidation pathway possibly due to a small contribution of NO2- reduction pathway. Further evaluation of isotopomer signatures of N2O using common enzymes of other processes related to N2O would improve the isotopomer analysis of N2O in various environments.

  8. Isotopomeric characterization of nitrous oxide produced by reaction of enzymes extracted from nitrifying and denitrifying bacteria

    T. Yamazaki

    2013-10-01

    Full Text Available Nitrous oxide (N2O is a potent greenhouse gas and produced in denitrification and nitrification in environmental nitrogen cycle by various microorganism. Site preference (SP of 15N in N2O, which is defined as the difference in the natural abundance of isotopomers 14N15NO and 15N14NO relative to 14N14NO, has been reported to be a useful tool to quantitatively distinguish N2O production pathway. To determine representative SP value for each microbial process, we firstly measured SP of N2O produced in the enzyme reaction of hydroxylamine oxidoreductase (HAO purified from two species of ammonia oxidizing bacteria (AOB, Nitrosomonas europaea and Nitrosococcus oceani, and that of nitric oxide reductase (NOR from Paracoccus denitrificans, respectively. The SP value for NOR reaction (−5.9 ± 2.1‰ showed nearly the same value as that reported for N2O produced by P. denitrificans in pure culture. In contrast, SP value for HAO reaction (36.3 ± 2.3‰ was a little higher than the values reported for N2O produced by AOB in aerobic pure culture. Using the SP values obtained by HAO and NOR reactions, we calculated relative contribution of the nitrite (NO2– reduction (which is followed by NO reduction to N2O production by N. oceani incubated under different O2 availability. Our calculations revealed that previous in vivo studies might have underestimated the SP value for NH2OH oxidation pathway possibly due to a small contribution of NO2– reduction pathway. Further evaluation of isotopomer signatures of N2O using common enzymes of other processes related to N2O would improve the isotopomer analysis of N2O in various environments.

  9. Environmental Development Plan: uranium enrichment

    This Environmental Development Plan identifies and examines the environmental, health, safety, and socioeconomic concerns and corresponding requirements associated with the DOE research, development, demonstration, and operation of the Uranium Enrichment program, including the gaseous diffusion process, the centrifuge process, centrifuge rotor fabrication, and related research and development activities

  10. Enrichment reduction for research reactors

    The worldwide activities on enrichment reduction for research reactors are reviewed and the national and international programs are described. Especially the following points are discussed: Benchmark calculations, reactor safety, fuel element development, irradiation tests, post irradiation examinations, full core demonstrations, activities of the GKSS and economical questions. (orig.)

  11. Real-time PCR assay for the simultaneous quantification of nitrifying and denitrifying bacteria in activated sludge

    Geets, J.; Cooman, M. de; Wittebolle, L.; Verstraete, W.; Boon, N. [Ghent Univ. (BE). Lab. of Microbial Ecology and Technology (LabMET); Heylen, K.; Vanparys, B.; Vos, P. de [Ghent Univ. (Belgium). Dept. of Biochemistry, Physiology and Microbiology

    2007-05-15

    In order to improve wastewater treatment processes, a need exists for tools that rapidly give detailed insight into the community structure of activated sludge, supplementary to chemical and physical data. In this study, the advantages of microarrays and quantitative polymerase chin reaction (PCR) methods were combined into a real-time PCR assay that allows the simultaneous quantification of phylogenetic and functional genes involved in nitrification and denitrification processes. Simultaneous quantification was possible along a 5-log dynamic range and with high linear correlation (R{sup 2}>0.98). The specificity of the assay was confirmed by cloning and sequencing analyses of PCR amplicons obtained from activated sludge. The real-time assay was validated on mixed liquid samples of different treatment plants, which varied in nitrogen removal rate. The abundance of ammonia oxidizers was in the order of magnitude of 10{sup 6} down to 10{sup 4} ml{sup -1}, whereas nitrite oxidizers were less abundant (10{sup 3}-10{sup 1} order of magnitude). The results were in correspondence with the nitrite oxidation rate in the sludge types. As for the nirS, nirK, and nosZ gene copy numbers, their abundance was generally in the order of magnitude of 10{sup 8}-10{sup 5}. When sludge samples were subjected to lab-scale perturbations, a decrease in nitrification rate was reflected within 18 h in the copy numbers of nitrifier genes (decrease with 1 to 5 log units), whereas denitrification genes remained rather unaffected. These results demonstrate that the method is a fast and accurate tool for the analysis of the (de)nitrifying community structure and size in both natural and engineered environmental samples. (orig.)

  12. Asymmetric reduction of ketones and β-keto esters by (S)-1-phenylethanol dehydrogenase from denitrifying bacterium Aromatoleum aromaticum.

    Dudzik, A; Snoch, W; Borowiecki, P; Opalinska-Piskorz, J; Witko, M; Heider, J; Szaleniec, M

    2015-06-01

    Enzyme-catalyzed enantioselective reductions of ketones and keto esters have become popular for the production of homochiral building blocks which are valuable synthons for the preparation of biologically active compounds at industrial scale. Among many kinds of biocatalysts, dehydrogenases/reductases from various microorganisms have been used to prepare optically pure enantiomers from carbonyl compounds. (S)-1-phenylethanol dehydrogenase (PEDH) was found in the denitrifying bacterium Aromatoleum aromaticum (strain EbN1) and belongs to the short-chain dehydrogenase/reductase family. It catalyzes the stereospecific oxidation of (S)-1-phenylethanol to acetophenone during anaerobic ethylbenzene mineralization, but also the reverse reaction, i.e., NADH-dependent enantioselective reduction of acetophenone to (S)-1-phenylethanol. In this work, we present the application of PEDH for asymmetric reduction of 42 prochiral ketones and 11 β-keto esters to enantiopure secondary alcohols. The high enantioselectivity of the reaction is explained by docking experiments and analysis of the interaction and binding energies of the theoretical enzyme-substrate complexes leading to the respective (S)- or (R)-alcohols. The conversions were carried out in a batch reactor using Escherichia coli cells with heterologously produced PEDH as whole-cell catalysts and isopropanol as reaction solvent and cosubstrate for NADH recovery. Ketones were converted to the respective secondary alcohols with excellent enantiomeric excesses and high productivities. Moreover, the progress of product formation was studied for nine para-substituted acetophenone derivatives and described by neural network models, which allow to predict reactor behavior and provides insight on enzyme reactivity. Finally, equilibrium constants for conversion of these substrates were derived from the progress curves of the reactions. The obtained values matched very well with theoretical predictions. PMID:25549618

  13. Effect of land use on the density of nitrifying and denitrifying bacteria in the Colombian Coffee Region

    Vallejo Quintero Victoria Eugenia

    2011-12-01

    Full Text Available

    Soil microbial communities involved in the cycling of nitrogen (N are essential to maintaining and improving soil fertility, productivity and functionality of natural and agricultural ecosystems. However, some compounds generated during the metabolic processes performed by nitrifying (NB and denitrifying (DB bacteria are associated with the production of greenhouse gases, groundwater pollution and acidification. Therefore, the study of these bacteria is essential for economic and environmental sustainability. This study evaluated the effect of different land uses in two river basins (La Vieja and Otun on NB and DB densities. Two sampling events (SE were conducted by selecting the most representative land uses. Physicochemical (T , pH, moisture and nitrate and microbiological properties (NB and DB densities were evaluated. In both SEs, significantly higher densities of NB and DB were observed in the land uses: pasture, guadua (DB only and unshaded coffee (La Vieja and onion (Otun. These land uses, excluding guadua, are dependent on nitrogen fertilizers, which together with the activities of grazing livestock on pastures may lead to greater availability of substrates for the NB. The use of agricultural machinery and overgrazing in pasture and onion uses generate compacted soil and other physical disturbances, encouraging the growth of DB. Forests had the lowest densities of NB and DB possibly due to a reduced availability of N and the releasing of allelopathic compounds from certain plants. Finally, the densities of ammonium-oxidizing bacteria had the greatest differences between the land uses evaluated, demonstrating its high sensitivity to agricultural management practices and livestock. We suggest that changes in the abundance of this community could serve as a relevant and cost-effective bioindicator for soil monitoring.

  14. Denitratimonas tolerans gen. nov., sp. nov., a denitrifying bacterium isolated from a bioreactor for tannery wastewater treatment.

    Han, Song-Ih; Kim, Ju-Ok; Lee, Ye-Rim; Ekpeghere, Kalu I; Koh, Sung-Cheol; Whang, Kyung-Sook

    2016-06-01

    A denitrifying bacterium, designated strain E4-1(T), was isolated from a bioreactor for tannery wastewater treatment, and its taxonomic position was investigated using a polyphasic approach. Strain E4-1(T), a facultative anaerobic bacterium, was observed to grow between 0 and 12 % (w/v) NaCl, between pH 3.0 and 12.0. Cells were found to be oxidase-positive and catalase-negative. Phylogenetic analyses based on 16S rRNA gene sequences showed that strain E4-1(T) forms a distinct lineage with respect to closely related genera in the family Xanthomonadaceae, and is closely related to Chiayiivirga, Aquimonas and Dokdonella, and the levels of 16S rRNA gene sequence similarity with respect to the type species of related genera are less than 93.9 %. The predominant respiratory quinone was determined to be ubiquinone-8 (Q-8) and the major cellular fatty acids were determined to be iso-C15:0, iso-C17:1 ω9c, iso-C11:0 and iso-C11:0 3OH. Based on physiological, biochemical and chemotaxonomic properties together with results of comparative 16S rRNA gene sequence analysis, strain E4-1(T) is considered to represent a novel species in a new genus, for which the name Denitratimonas tolerans gen. nov., sp. nov. is proposed. The type strain is E4-1(T) (=KACC 17565(T) = NCAIM B 025327(T)). PMID:27108138

  15. Selective enrichment of a methanol-utilizing consortium using pulp & paper mill waste streams

    Gregory R. Mockos; William A. Smith; Frank J. Loge; David N. Thompson

    2007-04-01

    Efficient utilization of carbon inputs is critical to the economic viability of the current forest products sector. Input carbon losses occur in various locations within a pulp mill, including losses as volatile organics and wastewater . Opportunities exist to capture this carbon in the form of value-added products such as biodegradable polymers. Waste activated sludge from a pulp mill wastewater facility was enriched for 80 days for a methanol-utilizing consortium with the goal of using this consortium to produce biopolymers from methanol-rich pulp mill waste streams. Five enrichment conditions were utilized: three high-methanol streams from the kraft mill foul condensate system, one methanol-amended stream from the mill wastewater plant, and one methanol-only enrichment. Enrichment reactors were operated aerobically in sequencing batch mode at neutral pH and 25°C with a hydraulic residence time and a solids retention time of four days. Non-enriched waste activated sludge did not consume methanol or reduce chemical oxygen demand. With enrichment, however, the chemical oxygen demand reduction over 24 hour feed/decant cycles ranged from 79 to 89 %, and methanol concentrations dropped below method detection limits. Neither the non-enriched waste activated sludge nor any of the enrichment cultures accumulated polyhydroxyalkanoates (PHAs) under conditions of nitrogen sufficiency. Similarly, the non-enriched waste activated sludge did not accumulate PHAs under nitrogen limited conditions. By contrast, enriched cultures accumulated PHAs to nearly 14% on a dry weight basis under nitrogen limited conditions. This indicates that selectively-enriched pulp mill waste activated sludge can serve as an inoculum for PHA production from methanol-rich pulp mill effluents.

  16. Observation of high seasonal variation in community structure of denitrifying bacteria in arable soil receiving artificial fertilizer and cattle manure by determining T-RFLP of nir gene fragments

    Priemé, Anders; Wolsing, Martin

    2004-01-01

    Temporal and spatial variation of communities of soil denitrifying bacteria at sites receiving mineral fertilizer (60 and 120 kg N ha-1 year-1) and cattle manure (75 and 150 kg N ha-1 year-1) were explored using terminal restriction fragment length polymorphism (T-RFLP) analyses of PCR amplified...... significant seasonal shift in the community structure of nirK-containing bacteria. Also, sites treated with mineral fertilizer or cattle manure showed different communities of nirK-containing denitrifying bacteria, since the T-RFLP patterns of soils treated with these fertilizers were significantly different...... investigated belonged to a yet uncultivated cluster of denitrifying bacteria....

  17. Community Structure Evolution and Enrichment of Glycogen-Accumulating Organisms Producing Polyhydroxyalkanoates from Fermented Molasses▿

    Pisco, Ana R.; Bengtsson, Simon; Werker, Alan; Reis, Maria A. M.; Lemos, Paulo C.

    2009-01-01

    An open mixed culture was enriched with glycogen-accumulating organisms (GAOs) by using a sequencing batch reactor and treating an agroindustrial waste (sugar cane molasses) under cyclic anaerobic-aerobic conditions. Over a 1-year operating period, the culture exhibited a very stable GAO phenotype with an average polyhydroxyalkanoate (PHA) content of 17% total suspended solids. However, the GAO microbial community evolved over the course of operation to a culture exhibiting unusual characteri...

  18. Removal of NO3--N from polluted groundwater in decommissioned mining area in an in-situ leach uranium mine by denitrifying bacteria bioreactor

    The pollution of groundwater by NO3- in the decommissioned mining area in an in-situ leach uranium mine is being paid more and more attention. The denitrifying bacteria by domesticating the sludge taken from the decommissioned mining area in an in-situ leach uranium mine in North West of China were obtained. An up-flow fixed-bed denitrifying bacteria bioreactor was designed. The effects of pH, the ratio of carbon to nitrogen and HRT on the removal of NO3--N from the polluted groundwater in the decommissioned mining area in the in-situ leach uranium mine by the denitrifying bacteria bioreactor were investigated. The results show that when pH, the concentration of NO3--N and HRT are set to 6.50, 1000 mg/L and 2.3 h, respectively, the removal of the NO3--N amounts to 97%, the NO3--N degradation rate is 388 mg/(h · L) and the capacity of the bioreactor is 0. 35 m3/(h · m3). When pH , the concentration of NO3--N and HRT are set to 6.50, 550 mg/L and 1.4 h, respectively, the removal of the NO3--N amounts to 96% and the capacity of the bioreactor is 0.62 m3/(h · m3). The bioreactor should work on the operation conditions that pH is 5.00-8.00 and the ratio of carbon to nitrogen is 0.6-0.8. (authors)

  19. Laboratory and field studies on BTEX biodegradation in a fuel-contaminated aquifer under denitrifying conditions. Book chapter, May 88-Dec 92

    Leaking underground storage tanks are a major source of groundwater contamination by petroleum hydrocarbons. Of the approximately 1.4 million underground tanks storing gasoline in the United States, some petroleum experts estimate that 75,000 to 100,000 are leaking (Feliciano 1984). Gasoline and other fuels contain benzene, toluene, ethylbenzene, and xylenes (collectively known as BTEX), which are hazardous compounds regulated by the U.S. Environmental Protection Agency (EPA 1977). Laboratory studies were conducted in conjunction with a field demonstration project on nitrate-mediated biorestoration of a fuel-contaminated aquifer at a U.S. Coast Guard facility in Traverse City, MI. Microcosms were prepared under either aerobic or strictly anaerobic, denitrifying conditions using core samples aseptically obtained from the aquifer. The microcosms were spiked with aromatic hydrocarbons (BTEX) and incubated as 12 C. Virtually all of the aromatic hydrocarbons, including benzene, were degraded to below detection limits within seven days under aerobic conditions, although o-xylene was somewhat more recalcitrant. Under denitrifying conditions, toluene, ethylbenzene, m-xylene, and 1,2,4-trimethylbenzene were also degraded to below detection limits, although this occurred between two to three weeks. o-Xylene was only slowly degraded and benzene was recalcitrant under denitrifying conditions. In the field demonstration project, an infiltration gallery was used to recirculate water at a rate sufficient to create a water table mound encompassing the contaminated interval. After hydraulic equilbrium was achieved nitrate and nutrients were added to the recharge water. Benzene removal occurred before nitrate addition; mass balances indicated that sufficient oxygen was recirculated to account for complete biodegradation aerobically

  20. Enrichment into the 21st century

    This article discusses the future of the enrichment services market into the next century. It is estimated that demand for enrichment services will reach 31 million SWU by the end of the century and remain constant for the following 10 years. The current world enrichment capacity is 44 million SWU, or some 50% ahead of the demand. This oversupply is projected to continue into the next century, but in spite of this, several suppliers are planning new enrichment facilities. HEU as a source of enriched uranium is examined. Overall, long-term prices for enrichment services are expected to decline in the coming decade

  1. Presence of Two Different Active nirS Nitrite Reductase Genes in a Denitrifying Thauera sp. from a High-Nitrate-Removal-Rate Reactor

    Etchebehere, Claudia; Tiedje, James

    2005-01-01

    The nirS nitrite reductase genes were studied in two strains (strains 27 and 28) isolated from two denitrifying reactors and characterized as Thauera according to their 16S rRNA gene sequences. Strain 28 contains a single nirS sequence, which is related to the nirS of Thauera mechernichensis, and strain 27 contains two nirS sequences; one is similar to the nirS sequence from Thauera mechernichensis (gene 2), but the second one (gene 8) is from a separate clade with nirS from Pseudomonas stutz...

  2. Draft genome sequence of the denitrifying strain Kiloniella sp. P1-1 isolated from the gut microflora of Pacific white shrimp, Litopenaeus vannamei.

    Wang, Liping; Li, Xiaoyi; Shao, Zongze

    2015-12-01

    A denitrifying bacterium, Kiloniella sp. P1-1, was isolated from the intestinal tract contents of Pacific white shrimp (Litopenaeus vannamei) from the Taiwan Strait, China. Here we report the draft genome sequence of Kiloniella sp. P1-1, which comprised 4,312,146 bp in 85 contigs with a G+C content of 46.09%. Related genes involved in the denitrification process could be identified in this genome. The genome sequence of Kiloniella sp. P1-1 may provide insights into the mechanism of denitrification. PMID:26303669

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

    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 attenuation of environmental contaminants. Understanding of diversity and physiology of anaerobic PAH degradation will contribute to remediation efforts of low-oxygen environments such as aquifers or river sediments.

  4. Observation of high seasonal variation in community structure of denitrifying bacteria in arable soil receiving artificial fertilizer and cattle manure by determining T-RFLP of nir gene fragments

    Priem, Anders; Wolsing, Martin

    2004-01-01

    Temporal and spatial variation of communities of soil denitrifying bacteria at sites receiving mineral fertilizer (60 and 120 kgNha-1year-1) and cattle manure (75 and 150 kgNha-1year-1) were explored using terminal restriction fragment length polymorphism (T-RFLP) analyses of PCR amplified...... significant seasonal shift in the community structure of nirK-containing bacteria. Also, sites treated with mineral fertilizer or cattle manure showed different communities of nirK-containing denitrifying bacteria, since the T-RFLP patterns of soils treated with these fertilizers were significantly different...

  5. Sequential enrichment of microbial population exhibiting enhanced biodegradation of crude oil

    The distribution of oil-degrading bacteria in the coastal waters and sediments of Hokkaido, Japan, was surveyed. It was found that the potential of mixed microbial populations to degrade weathered crude oil was not confined to any ecological components (water or sediment) nor to the sampling stations. One microbial culture that was stable during repeated subculturing degraded 45% of the saturates and 20% of the aromatics present in crude oil in 10 days during the initial screening. The residual hydrocarbons in this culture were extracted by chloroform and dispersed in a fresh seawater-based medium and subsequently inoculated with microorganisms from the first culture. After full growth of the second culture, the residual hydrocarbons were extracted and dispersed in a fresh medium in which microorganisms from the second culture had been inoculated. This sequential process was carried out six times to enrich those microorganisms that grew on the recalcitrant components of crude oil. After repeated exposure of the residual crude oil to the enriched microorganisms, about 80% of the initially added crude oil was degraded. The cultures obtained after each enrichment cycle were kept, and the degradation of fresh crude oil by the enriched microorganisms was monitored. The degrading activity of the enriched cultures increased as the number of enrichment cycles increased. A microbial population that had been selected six times on the residual crude oil could degrade 70% of the saturates and 30% of the aromatics of crude oil, indicating that growth of a microbial population on residual crude oil improved its ability to biodegrade crude oil. 21 refs., 2 tabs., 7 figs

  6. Boron Enrichment in Martian Clay

    Stephenson, James D.; Hallis, Lydia J.; Nagashima, Kazuhide; Freeland, Stephen J.

    2013-01-01

    We have detected a concentration of boron in martian clay far in excess of that in any previously reported extra-terrestrial object. This enrichment indicates that the chemistry necessary for the formation of ribose, a key component of RNA, could have existed on Mars since the formation of early clay deposits, contemporary to the emergence of life on Earth. Given the greater similarity of Earth and Mars early in their geological history, and the extensive disruption of Earth's earliest minera...

  7. Uranium enrichment and public policy

    Alternative policies to the present government monopoly of the uranium enrichment process are discussed in this monograph. Alternative policies analyzed are continued government monopoly and operation by the Department of Energy, establishment of a government corporation, and complete relinquishing of government control with the private sector taking the risks of constructing additional capacity. The material is divided into 6 chapters. Chapter 1 describes the fuel cycle and the public policy issues involved. The enrichment process, current capacity, and the operation of the plants by the government are described in Chapter 2. Chapter 3 estimates future demand for electricity in the U.S. and forecasts the demand for nuclear power and for enrichment services. Chapter 4 describes the historical background, including the Nuclear Fuel Assurance Act, opposition to it, and its demise. The basic policy options the government faces with an analysis of the advantages and disadvantages of each are included in Chapter 5. The final chapter summarizes the findings and presents the conclusion that, from an economic efficiency viewpoint, selling the existing plants and turning the industry over to the private sector would be best. An appendix describes some of the issues involved in plutonium recycling

  8. Evaluation of vitamin C-enriched Artemia nauplii for larvae of the giant freshwater prawn

    Merchie, G.; Lavens, P.; Radull, J.; Nelis, H.; De Leenheer, A. P.; Sorgeloos, P.

    2000-01-01

    The effects of high levels of ascorbic acid (AA) delivered through enriched live food has been verified through the successful culture of larval giant freshwater prawn, Macrobrachium rosenbergii Two successive feeding trials were set up using a control (550 g AA g-1 DW) and two different AA-enrichment levels in Artemia (1300 and 2750 g AA g-1 DW). Under standard culture conditions, no differences in growth nor survival could be observed demonstrating that the nutritional requirements are be...

  9. Uranium enrichment (a strategy analysis overview)

    An analysis of available information on enrichment technology, separative work supply and demand, and SWU cost is presented. Estimates of present and future enrichment costs are provided for use in strategy analyses of alternate nuclear fuel cycles and systems. (auth)

  10. Lysine-Enriched Cecropin-Mellitin Antimicrobial Peptides with Enhanced Selectivity ▿

    Sato, Hiromi; Feix, Jimmy B

    2008-01-01

    Lysine-enriched analogs of the cecropin-mellitin hybrid peptide, CA1-7 M2-9 (designated CM15), designed with optimized amphipathicity, retained antimicrobial activities similar to that of wild-type CM15 and had substantially reduced levels of hemolytic activity and cytotoxicity toward cultured macrophages, resulting in enhanced selectivity. These lysine-enriched analogs provide templates for improved CM15 peptide or peptidomimetic antibiotics.

  11. Enriched uranium sales: effect on supply industry

    The subject is covered in sections: introduction (combined effect of low-enriched uranium (LEU) inventory sales and utility services enrichment contract terms); enrichment market overview; enrichment market dynamics; the reaction of the US Department of Energy; elimination of artificial demand; draw down of inventories; purchase and sale of LEU inventories; tails assay option; unfulfilled requirements for U3O8; conclusions. (U.K.)

  12. Enriching Music and Language Arts Experiences

    Flohr, John W.

    2006-01-01

    The article focuses on enriching music and language arts experiences of students. Music can enrich literature and language arts, poetry, theater arts, transitions, science, and math, as well as help meet special learner needs. A well-understood example of enrichment is the alphabet song. A music or classroom teacher using the alphabet song helps…

  13. Urenco's contribution to the enrichment market

    The paper deals with the Present status of Urenco Enrichment Plants and Order Book, Urenco's future development, The Western World enrichment supply situation, Urenco's Policy on the reliability of enrichment supplies, and its contribution to a reliable, stable and competitive world enrichment market. Urenco's current plant capacity and forward committed expansion plans up to 1990 and its present position in the western world enrichment market are described. The application of Urenco's centrifuge technology to the enrichment of recycles uranium and the relevance of laser enrichment development to Urenco are discussed. The paper next discusses the structure and main characteristics of the major sectional markets of the western world enrichment market, i.e. particularly in the United States, Western Europe and Asia (Japan, Korea and Taiwan). Urenco's contractual approach regarding the reliability of enrichment supplies is discussed, it is shown that Urenco's development of the centrifuge process to the point of economic maturity, Urenco's establishment of a multi-national commercial enrichment organization, Urenco's achievement in the world enrichment market by a commercial and competitive approach, Urenco's preparedness to negotiate suitable supply arrangements including the establishment of plants in other market areas have vastly contributed to a stabilisation of the world enrichment market. (J.P.N.)

  14. Bioaugmentation of nitrate-dependent anaerobic ferrous oxidation by heterotrophic denitrifying sludge addition: A promising way for promotion of chemoautotrophic denitrification.

    Wang, Ru; Zheng, Ping; Zhang, Meng; Zhao, He-Ping; Ji, Jun-Yuan; Zhou, Xiao-Xin; Li, Wei

    2015-12-01

    Nitrate-dependent anaerobic ferrous oxidation (NAFO) is a new and valuable bio-process for the treatment of wastewaters with low C/N ratio, and the NAFO process is in state of the art. The heterotrophic denitrifying sludge (HDS), possessing NAFO activity, was used as bioaugmentation to enhance NAFO efficiency. At a dosage of 6% (V/V), the removal of nitrate and ferrous was 2.4 times and 2.3 times of as primary, and the volumetric removal rate (VRR) of nitrate and ferrous was 2.4 times and 2.2 times of as primary. Tracing experiments of HDS indicated that the bioaugmentation on NAFO reactor was resulted from the NAFO activity by HDS itself. The predominant bacteria in HDS were identified as Thauera (52.5%) and Hyphomicrobium (20.0%) which were typical denitrifying bacteria and had potential ability to oxidize ferrous. In conclusion, HDS could serve as bioaugmentation or a new seeding sludge for operating high-efficiency NAFO reactors. PMID:26348287

  15. Two-step nitrification in a pure moving bed biofilm reactor-membrane bioreactor for wastewater treatment: nitrifying and denitrifying microbial populations and kinetic modeling.

    Leyva-Díaz, J C; González-Martínez, A; Muñío, M M; Poyatos, J M

    2015-12-01

    The moving bed biofilm reactor-membrane bioreactor (MBBR-MBR) is a novel solution to conventional activated sludge processes and membrane bioreactors. In this study, a pure MBBR-MBR was studied. The pure MBBR-MBR mainly had attached biomass. The bioreactor operated with a hydraulic retention time (HRT) of 9.5 h. The kinetic parameters for heterotrophic and autotrophic biomasses, mainly nitrite-oxidizing bacteria (NOB), were evaluated. The analysis of the bacterial community structure of the ammonium-oxidizing bacteria (AOB), NOB, and denitrifying bacteria (DeNB) from the pure MBBR-MBR was carried out by means of pyrosequencing to detect and quantify the contribution of the nitrifying and denitrifying bacteria in the total bacterial community. The relative abundance of AOB, NOB, and DeNB were 5, 1, and 3%, respectively, in the mixed liquor suspended solids (MLSS), and these percentages were 18, 5, and 2%, respectively, in the biofilm density (BD) attached to carriers. The pure MBBR-MBR had a high efficiency of total nitrogen (TN) removal of 71.81±16.04%, which could reside in the different bacterial assemblages in the fixed biofilm on the carriers. In this regard, the kinetic parameters for autotrophic biomass had values of YA=2.3465 mg O2 mg N(-1), μm, A=0.7169 h(-1), and KNH=2.0748 mg NL(-1). PMID:26264139

  16. Hydrogenotrophic denitrification process efficiency and the number of denitrifying bacteria (MPN) in the sequencing batch biofilm reactor (SBBR) with platinum and carbon anodes.

    Kłodowska, Izabella; Rodziewicz, Joanna; Janczukowicz, Wojciech; Gotkowska-Płachta, Anna; Cydzik-Kwiatkowska, Agnieszka

    2016-04-15

    This work reports on the effect of electric current density and anode material (platinum, carbon) on the concentration of oxidized and mineral forms of nitrogen, on physical parameters (pH, redox potential, electrical conductivity) and the number of denitrifying bacteria in the biofilm (MPN). Experiments were conducted under anaerobic conditions without and with the flow of electric current (with density of 79 mA·m(-2) and 132 mA·m(-2)). Results obtained in the study enabled concluding that increasing density of electric current caused a decreasing concentration of nitrate in the reactor with platinum anode (R1) and carbon anode (R2). Its concentration depended on anode material. The highest hydrogenotrophic denitrification efficiency was achieved in R2 in which the process was aided by inorganic carbon (CO2) that originated from carbon anode oxidation and the electrical conductivity of wastewater increased as a result of the presence of HCO3(-) and CO3(2-) ions. Strong oxidizing properties of the platinum anode (R1) prevented the accumulation of adverse forms of nitrogen, including nitrite and ammonia. The increase in electric current density affected also a lower number of denitrifying bacteria (MPN) in the biofilm in both reactors (R1 and R2). Metal oxides accumulated on the surface of the cathode had a toxic effect upon microorganisms and impaired the production of a hydrogen donor. PMID:26809836

  17. Nitrous oxide nitrification and denitrification 15N enrichment factors from Amazon forest soils.

    Prez, Tibisay; Garcia-Montiel, Diana; Trumbore, Susan; Tyler, Stanley; de Camargo, Plnio; Moreira, Marcelo; Piccolo, Marisa; Cerri, Carlos

    2006-12-01

    The isotopic signatures of 15N and 18O in N2O emitted from tropical soils vary both spatially and temporally, leading to large uncertainty in the overall tropical source signature and thereby limiting the utility of isotopes in constraining the global N2O budget. Determining the reasons for spatial and temporal variations in isotope signatures requires that we know the isotope enrichment factors for nitrification and denitrification, the two processes that produce N2O in soils. We have devised a method for measuring these enrichment factors using soil incubation experiments and report results from this method for three rain forest soils collected in the Brazilian Amazon: soil with differing sand and clay content from the Tapajos National Forest (TNF) near Santarm, Par, and Nova Vida Farm, Rondnia. The 15N enrichment factors for nitrification and denitrification differ with soil texture and site: -111 per thousand +/- 12 per thousand and -31 per thousand +/- 11 per thousand for a clay-rich Oxisol (TNF), -102 per thousand +/- 5 per thousand and -45 per thousand +/- 5 per thousand for a sandier Ultisol (TNF), and -10.4 per thousand +/- 3.5 per thousand (enrichment factor for denitrification) for another Ultisol (Nova Vida) soil, respectively. We also show that the isotopomer site preference (delta15Nalpha - delta15Nbeta, where alpha indicates the central nitrogen atom and beta the terminal nitrogen atom in N2O) may allow differentiation between processes of production and consumption of N2O and can potentially be used to determine the contributions of nitrification and denitrification. The site preferences for nitrification and denitrification from the TNF-Ultisol incubated soils are: 4.2 per thousand +/- 8.4 per thousand and 31.6 per thousand +/- 8.1 per thousand, respectively. Thus, nitrifying and denitrifying bacteria populations under the conditions of our study exhibit significantly different 15N site preference fingerprints. Our data set strongly suggests that N2O isotopomers can be used in concert with traditional N2O stable isotope measurements as constraints to differentiate microbial N2O processes in soil and will contribute to interpretations of the isotopic site preference N2O values found in the free troposphere. PMID:17205894

  18. Relationship between N2O Fluxes from an Almond Soil and Denitrifying Bacterial Populations Estimated by Quantitative PCR

    Matiasek, M.; Suddick, E. C.; Smart, D. R.; Scow, K. M.

    2008-12-01

    Cultivated soils emit substantial quantities of nitrous oxide (N2O), a greenhouse gas with almost 300 times the radiative forcing potential of CO2. Agriculture-related activities generate from 6 to 35 Tg N2O-N per year, or about 60 to 70% of global production. The microbial processes of nitrification, denitrification and nitrifier denitrification are major biogenic sources of N2O to the atmosphere from soils. Denitrification is considered the major source of N2O especially when soils are wet. The microbial N transformations that produce N2O depend primarily on nitrogen (N) fertilizer, with water content, available carbon and soil temperature being secondary controllers. Despite the fact that microbial processes are responsible for N2O emissions, very little is known about the numbers or types of populations involved. The objective of this study was to relate changes in denitrifying population densities, using quantitative PCR (qPCR) of functional genes, to N2O emissions in a fertilized almond orchard. Quantitative PCR targeted three specific genes involved in denitrification: nirS, nirK and nosZ. Copy numbers of the genes were related back to population densities and the portion of organisms likely to produce nitrous oxide. The study site, a 21.7 acre almond orchard fitted with micro-sprinklers, was fertigated (irrigated and fertilized simultaneously) with 50 lbs/acre sodium nitrate in late March 2008, then irrigated weekly. Immediately after the initial fertigation, fluxes of N2O and CO2, moisture content, inorganic N and denitrification gene copy numbers were measured 6 times over 24 days. Despite the fact that N2O emissions increased following fertigation, there was no consistent increase in any of the targeted genes. The genes nirK and nirS ranged from 0.4-1.4 107 and 0.4-1.4 108, whereas nosZ ranged from 2-8 106 copy numbers per g soil, respectively. Considerable variation, compounded by the small sample sizes used for DNA analysis, made it difficult to discern trends over time. High spatial variability was also observed with one of the field replicates have a substantially higher flux of N2O. This replicate also had the highest water filled pore space (WFPS) and water content, factors that likely favored denitrification. Water saturation of soil air space, optimal at >60% for denitrification to occur, was relatively low in the other field replicates. Thus, the low N2O flux measurements and gene copy numbers agreed in supporting the hypothesis denitrification was relatively low under the environmental conditions of these particular almond soils.

  19. Identification of the function of extracellular polymeric substances (EPS) in denitrifying phosphorus removal sludge in the presence of copper ion.

    Wang, Yayi; Qin, Jian; Zhou, Shuai; Lin, Ximao; Ye, Liu; Song, Chengkang; Yan, Yuan

    2015-04-15

    Industrial wastewater containing heavy metals that enters municipal wastewater treatment plants inevitably has a toxic impact on biological treatment processes. In this study, the impact of Cu(II) (0, 1.5, 2, 2.5, 3 mg/L) on the performance of denitrifying phosphorus removal (DPR) and microbial community structures was investigated. Particularly, the dynamic change in the amount and composition of extracellular polymeric substances (EPS), and the role of EPS in P removal, were assessed using three-dimensional excitation-emission matrix fluorescence spectroscopy combined with parallel factor (PARAFAC) analysis. The results showed that, after long-term adjustment, the P removal efficiency was maintained at 95 2.7% at Cu(II) addition up to 2.5 mg/L, but deteriorated when the Cu(II) addition was 3 mg/L. The EPS content, including proteins and humic substances, increased with increasing Cu(II) additions at concentrations ?2.5 mg/L. This property of EPS was beneficial for protecting phosphate-accumulating organisms (PAOs) against heavy metals, as both proteins and humic substances are strong ligands for Cu(II). Therefore, the PAOs abundance was still relatively high (67 3%) when Cu(II) accumulation in sludge was up to 10 mg/g SS. PARAFAC confirmed that aromatic proteins could be transformed into soluble microbial byproduct-like material when microorganisms were subjected to Cu(II) stress, owing to their strong metal ion complexing capacity. The increase in the percentage of humic-like substances enhanced the detoxification function of the sludge EPS. EPS accounted for approximately 26-47% of P removed by adsorption when Cu(II) additions were between 0 and 2.5 mg/L. The EPS function, including binding toxic heavy metals and P storage, enhanced the operating stability of DPR systems. This study provides us with a better understanding of (1) the tolerance of DPR sludge to copper toxicity and (2) the function of sludge EPS in the presence of heavy metals in biological P removal systems. PMID:25697691

  20. Enriching Orphans’ Potentials through Interpersonal and Intrapersonal Intelligence Enrichment Activities

    Nurulwahida Hj Azid

    2016-01-01

    Full Text Available Orphans are considered a minority and they should be given a greater emphasis so that they do not feel left out and can build their own lives without a sense of humility. This does not mean that the orphans should be pampered instead they should be given the confidence and motivation to strive for success in later life. Humility among orphans can be associated with interpersonal and intrapersonal intelligences. This study aims to evaluate the impact of problem-solving activity treatment based on the interpersonal and intrapersonal intelligences. 46 students from two orphanages were involved as the treatment group. The research design used was a one-group pretest-posttest design applied through a combination of quantitative and qualitative approaches. Enrichment activities that provided interpersonal and intrapersonal skills as evidenced in this study should be carried out regularly at orphanages. Our study has proven that orphans‟ rights to learn cannot be neglected and „no child left behind „policy needs to be carried through by everybody involved with orphans‟ well-being. Teachers and carers need to be trained to use these enrichment activities at their orphanages to help maximize the orphans‟ potentials.

  1. Trace elements affect methanogenic activity and diversity in enrichments from subsurface coal bed produced water

    KlausNüsslein

    2012-05-01

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

  2. Enrichment planting without soil treatment

    Hagner, Mats

    1998-12-31

    Where enrichment planting had been carried out with either of the two species Picea abies and Pinus contorta, the survival of the planted seedlings was at least as good as after planting in a normal clear cut area treated with soil scarification. This was in spite of the fact that the seedlings were placed shallow in the humus layer without any soil treatment. However, they were sheltered from insects by treatment before planting. Where enrichment planting was carried out with Pinus sylvestris the survival in dense forest was poor, but in open forest the survival was good. The growth of planted seedlings was enhanced by traditional clearing and soil treatment. However, this was for Pinus sylvestris not enough to compensate for the loss of time, 1-2 years, caused by arrangement of soil scarification. The growth of seedlings planted under crown cover was directly related to basal area of retained trees. However, the variation in height growth among individual seedlings was very big, which meant that some seedlings grow well also under a fairly dense forest cover. The pioneer species Pinus sylvestris reacted more strongly to basal area of retained trees than did the shade tolerant species Picea abies. Enrichment planting seems to be a necessary tool for preserving volume productivity, at places where fairly intensive harvest of mature trees has been carried out in stands of ordinary forest type in central Sweden. If double seedlings, with one Picea abies and one Pinus sylvestris, are used, the probability for long term establishment is enhanced 13 refs, 20 figs, 4 tabs

  3. Nematode Indicators of Organic Enrichment

    Ferris, Howard; Bongers, Tom

    2006-01-01

    The organisms of the soil food web, dependent on resources from plants or on amendment from other sources, respond characteristically to enrichment of their environment by organic matter. Primary consumers of the incoming substrate, including bacteria, fungi, plant-feeding nematodes, annelids, and some microarthropods, are entry-level indicators of enrichment. However, the quantification of abundance and biomass of this diverse group, as an indicator of resource status, requires a plethora of extraction and assessment techniques. Soluble organic compounds are absorbed by bacteria and fungi, while fungi also degrade more recalcitrant sources. These organisms are potential indicators of the nature of incoming substrate, but current methods of biomass determination do not reliably indicate their community composition. Guilds of nematodes that feed on bacteria (e.g., Rhabditidae, Panagrolaimidae) and fungi (e.g., Aphelenchidae, Aphelenchoididae) are responsive to changes in abundance of their food. Through direct herbivory, plant-feeding nematodes (e.g., many species of Tylenchina) also contribute to food web resources. Thus, analysis of the nematode community of a single sample provides indication of carbon flow through an important herbivore channel and through channels mediated by bacteria and fungi. Some nematode guilds are more responsive than others to resource enrichment. Generally, those bacterivores with short lifecycles and high reproductive potential (e.g., Rhabditidae) most closely mirror the bloom of bacteria or respond most rapidly to active plant growth. The feeding habits of some groups remain unclear. For example, nematodes of the Tylenchidae may constitute 30% or more of the individuals in a soil sample; further study is necessary to determine which resource channels they portray and the appropriate level of taxonomic resolution for this group. A graphic representation of the relative biomass of bacterivorous, fungivorous, and herbivorous nematodes provides a useful tool for assessing the importance of the bacterial, fungal, and plant resource channels in an extant food web. PMID:19259424

  4. EMIS: Enrichment Management Information System

    The objective of the effort was the development of a calculational tool which would permit DOE to make realistic projections of the separative work demands of its customers as an aid in planning for the future operation of the enrichment facilities. The calculational methods were to recognize the flexibility in scheduling, etc., available to DOE customers under their enrichment contreacts as well as the ability to pool separative work withdrawn under two or more contracts. It was also to be designed to permit the user to add new customers or delete existing contracts as well as to allow variation of the estimated start-up dates for those reactors currently under construction. The output of the model was to display projections of separative work demand as SWU and in triplet form for several categories of reactors. These included, in addition to the total demand on the enrichment complex, separate totals for domestic and foreign customers, for reactors with Requirements and Adjustable Fixed Commitment contracts and for reactors in operation, under construction and projected new reactors designated by the user. The foundations upon which the system is based are described in this report. This includes a discussion of the available alternatives for customer management of separative work and the assumptions and guidelines established for development of the model. The report also contains a description of the computational procedures used in the model, and a description of the EMIS system itself, which also serves as a users' manual for the code. Finally, samples demonstrating the use of the system and indicating the form of the input and output are included

  5. Boron enrichment in martian clay.

    Stephenson, James D; Hallis, Lydia J; Nagashima, Kazuhide; Freeland, Stephen J

    2013-01-01

    We have detected a concentration of boron in martian clay far in excess of that in any previously reported extra-terrestrial object. This enrichment indicates that the chemistry necessary for the formation of ribose, a key component of RNA, could have existed on Mars since the formation of early clay deposits, contemporary to the emergence of life on Earth. Given the greater similarity of Earth and Mars early in their geological history, and the extensive disruption of Earth's earliest mineralogy by plate tectonics, we suggest that the conditions for prebiotic ribose synthesis may be better understood by further Mars exploration. PMID:23762242

  6. Producing deuterium-enriched products

    A method of producing an enriched deuterium product from a gaseous feed stream of mixed hydrogen and deuterium, comprises: (a) combining the feed stream with gaseous bromine to form a mixture of the feed stream and bromine and exposing the mixture to an electrical discharge effective to form deuterium bromide and hydrogen bromide with a ratio of D/H greater than the ratio of D/H in the feed stream; and (b) separating at least a portion of the hydrogen bromide and deuterium bromide from the mixture. (author)

  7. Using metagenomic analyses to estimate the consequences of enrichment bias for pathogen detection

    Pettengill James B

    2012-07-01

    Full Text Available Abstract Background Enriching environmental samples to increase the probability of detection has been standard practice throughout the history of microbiology. However, by its very nature, the process of enrichment creates a biased sample that may have unintended consequences for surveillance or resolving a pathogenic outbreak. With the advent of next-generation sequencing and metagenomic approaches, the possibility now exists to quantify enrichment bias at an unprecedented taxonomic breadth. Findings We investigated differences in taxonomic profiles of three enriched and unenriched tomato phyllosphere samples taken from three different tomato fields (n = 18. 16S rRNA gene meteganomes were created for each of the 18 samples using 454/Roche’s pyrosequencing platform, resulting in a total of 165,259 sequences. Significantly different taxonomic profiles and abundances at a number of taxonomic levels were observed between the two treatments. Although as many as 28 putative Salmonella sequences were detected in enriched samples, there was no significant difference in the abundance of Salmonella between enriched and unenriched treatments. Conclusions Our results illustrate that the process of enriching greatly alters the taxonomic profile of an environmental sample beyond that of the target organism. We also found evidence suggesting that enrichment may not increase the probability of detecting a target. In conclusion, our results further emphasize the need to develop metagenomics as a validated culture independent method for pathogen detection.

  8. Marketing keeps enrichment market alive

    A resurgence of interest by utilities in the uranium market has occured, as a result of marketing efforts by primary suppliers -especially the US DoE and Russia's Techsnabexport (TENEX). Price, contract flexibility and reliability of supply have been the dominant factors governing utility purchase considerations. Many US utilities have sought to diversify their enrichment supply sources for economic reasons. Low spot market prices have encouraged major suppliers to be more competitive and to offer attractive prices in the long-term market. Although Eurodif and Urenco are at a disadvantage because of exchange rates, all of the primary suppliers have focused on the post-1995 period -especially in the USA where large unfulfilled requirements remain. The DoEs success in capturing part of this unfilled demand proves it can be competitive. The emergence and success of TENEX as a supplier indicates buyer willingness to explore new supply sources. However, the future success of all enrichment suppliers depends on their ability to adapt to a changing market by reducing excess capacity, controlling costs and appealing to utilities with competitive contracts. (Author)

  9. Urine culture

    Culture and sensitivity - urine ... when urinating. You also may have a urine culture after you have been treated for an infection. ... when bacteria or yeast are found in the culture. This likely means that you have a urinary ...

  10. Evaluación del crecimiento de un cultivo de Daphnia magna alimentado con Saccharomyces cereviseae y un enriquecimiento con avena soya Avaliação do crescimento de um cultivo de Daphnia magna alimentada con Saccharomyces cereviseae e um meio enrriquecido com aveia soja Growth culture evaluation of Daphnia magna feed with Saccharomyces cereviseae enrichment with oat soy

    Lucía E Ocampo

    2010-03-01

    Full Text Available Se realizó el cultivo experimental de cladócero Daphnia magna alimentado con probióticos. Se realizaron 16 ensayos experimentales, en el laboratorio de larvicultura de peces y alimento vivo (Larpeali de la Facultad de Ciencias Agrarias de la Universidad de Antioquia, bajo condiciones controladas de temperatura ambiente (21 - 25 °C, temperatura del agua (22 - 23 °C y pH (7.6. Empleando una dieta de Saccharomyces cereviseae y un medio de enriquecimiento con ácidos grasos (n-6 proveniente de harina avena-soya. Las concentraciones de dieta y enriquecimiento fueron de 25 ppm y 12.5 ppm, en arreglo factorial 2x2 (2 niveles de dieta con levadura y 2 niveles dieta con avena de soya, los cultivos de Daphnia por tratamiento se realizaron con cuatro replicas con el fin de determinar su efecto sobre el desempeño de la población. Se alimentaron cada tercer día, durante 15 días, evaluándose el número de organismos al final del período. Se obtuvieron diferencias altamente significativas (p0.05. Se evidenció que la combinación de estos componentes en sus concentraciones más altas potenció el crecimiento de la Daphnia magna, alcanzando un número de microcrustáceos de 826ª Daphnias/L ± 9.57. Se puede concluir que los cladóceros por sus características de crecimiento en cultivo, presentan adaptación favorable a las condiciones de manejo para la producción de biomasa útil como alimento vivo en acuicultura.O objetivo do experimento foi avaliar o efeito das gorduras saturadas e insaturadas e seu grau de proteção (protegidas e sem proteção sob a cinética e a extensão da degradação in vitro da matéria seca (MS em dietas para ruminantes. Quatro dietas foram formuladas para conter o mesmo nível de energia (3200 Kcal ED /Kg MS e proteína (13%PC a ração total misturada (RTM, de acordo com as necessidades nutricionais de ovelhas em fase de finalização, quatro tipos de gorduras foram feitas com 8% de MS e foram testados tipos de gorduras foram testados: 1 gordura saturada sem proteção (GSSP, 2 gordura saturada protegida (GSP, 3 gordura insaturada sem proteção (GISP e 4 gordura insaturada protegida (GIP. Para estimar a cinética e a extensão da degradação da MS e cinética da produção de gás, as rações foram avaliadas mediante a técnica in vitro de produção de gás, seguindo os modelos propostos por Orskov e McDonald (1979 y France et al (1993, para isto foi empregado o procedimento PROC NLIN de SAS (2001. Ao mesmo tempo, foi realizado um analise de medidas repetidas no tempo para verificar o efeito dos tratamentos sobre a produção de gás e a degradação da MS no tempo com ajuda do procedimento PROC MIXED do SAS (2001. Neste experimento verificou-se que a utilização de gorduras insaturadas protegidas não afetou a degradação de MS quando comparadas com as não protegidas. No caso das gorduras saturadas, não houve um claro efeito da proteção sob a degradação e a cinética de fermentação da MS.Experimental cultivate cladocerans Daphnia magna feed with probiotics was realized. At University of Antioquia, Faculty of Agrarian Science, Larviculture, fish and food live Laboratory (Larpeali, under controlled conditions of environmental temperature between (21 - 25 °C, water temperature (22 - 23°C and pH (7.6, were performed. One diet of Saccharomyces cereviseae was used and one enriched medium of oat-soy flour (fatty acid, n-6 to concentrations of diet and enriched medium were the same, 25 ppm and 12.5 ppm, 2x2 factorial treatment arrangements (2 levels of Saccharomyces cereviseae and 2 levels of oat-soy flour, diets was fed to four replicates of Daphnia culture per treatment in order to determine their effects on the populations cultivated. A 15-days feeding, each third day, trial were conducted, to assess the organisms at final period. The results showed significant high difference (p0.05. The results showed that the combination of their high concentrations of components enhanced the population growth of Daphnia magna, reaching a number of microcrustaceans of 826ª ± 9.57. These cladocerans for their characteristics of growth in culture, present favorable adaptation to the handling conditions to produce biomasses potentially useful as nutritious particle as live food in aquaculture purposes.

  11. Safeguards Culture

    Frazar, Sarah L.; Mladineo, Stephen V.

    2012-07-01

    The concepts of nuclear safety and security culture are well established; however, a common understanding of safeguards culture is not internationally recognized. Supported by the National Nuclear Security Administration, the authors prepared this report, an analysis of the concept of safeguards culture, and gauged its value to the safeguards community. The authors explored distinctions between safeguards culture, safeguards compliance, and safeguards performance, and evaluated synergies and differences between safeguards culture and safety/security culture. The report concludes with suggested next steps.

  12. Organizational Culture

    Adrian HUDREA

    2006-02-01

    Full Text Available Cultural orientations of an organization can be its greatest strength, providing the basis for problem solving, cooperation, and communication. Culture, however, can also inhibit needed changes. Cultural changes typically happen slowly – but without cultural change, many other organizational changes are doomed to fail. The dominant culture of an organization is a major contributor to its success. But, of course, no organizational culture is purely one type or another. And the existence of secondary cultures can provide the basis for change. Therefore, organizations need to understand the cultural environments and values.

  13. Microscale effects on denitrification: does the ability of denitrifying bacteria to reduce N2O depend on their position in the soil matrix?

    Dörsch, Peter; Nadeem, Shahid; Almås, Åsgeir; Bakken, Lars R.

    2013-04-01

    Soil is a heterogeneous matrix with a variety of microhabitats which probably select for organisms with distinct functional traits. The composition and functioning of soil denitrifier communities (DC) has been studied intensely over the last decades, primarily because of their role in the emission of N2O from soil. The tacit assumption in such studies is that the soil microbial community is one "thing". In the present study, we challenge the concept of DC as a homogenous entity and suggest a stratification of denitrifier function based on the position within the soil matrix. We hypothesize that soil contains "inner" and "outer" habitats; the inner consisting of sites within crevices and cavities of the soil mineral material, structured organic materials and strong biofilms, the outer consisting of exposed surfaces and macropores within the soil matrix. We further believe that sequential dispersion/extraction by density gradient centrifugation (DGC) can be used to crudely separate the organisms residing in the two habitat types. We operationally define loosely attached cells (LAC) as those which are liberated from soil particles by moderate dispersion of soils. LAC were separated from the soil by DGC, and the pellets at the bottom of the gradients containing bacteria still attached to or embedded in soil material were then subjected to a stronger dispersion to release the more strongly attached cells (SAC) which were again separated from the soil by DGC. We hypothesized that SAC are cells situated deeper in biofilms or other protective structures within the soil matrix than LAC. We further hypothesized that the two habitats select for different characteristics regarding the regulation of denitrification. In short, SAC were expected to express N2O-reductase earlier than LAC, because SAC experience anoxia and lack of NOx more frequently than LAC. First results from incubations with LAC and SAC from different soils lend strong support to this hypothesis; in response to oxygen depletion, both denitrifying communities expressed similar apparent growth rates, both produced a mixture of NO, N2O and N2, but the relative amount of N2O produced was much higher for LAC than SAC. This reemphasizes the significance of soil structure, habitable pore space, predation and bacteria-mineral interactions as regulating factors for N2O emission in mineral soils and has implications for the understanding of selection pressures for sustained N2O-reductase activity in soil.

  14. Nitrite survival and nitrous oxide production of denitrifying phosphorus removal sludges in long-term nitrite/nitrate-fed sequencing batch reactors.

    Wang, Yayi; Zhou, Shuai; Ye, Liu; Wang, Hong; Stephenson, Tom; Jiang, Xuxin

    2014-12-15

    Nitrite-based phosphorus (P) removal could be useful for innovative biological P removal systems where energy and carbon savings are a priority. However, using nitrite for denitrification may cause nitrous oxide (N2O) accumulation and emissions. A denitrifying nitrite-fed P removal system [Formula: see text] was successfully set up in a sequencing batch reactor (SBR) and was run for 210 days. The maximum pulse addition of nitrite to [Formula: see text] was 11 mg NO2(-)-N/L in the bulk, and a total of 34 mg NO2(-)-N/L of nitrite was added over three additions. Fluorescent in situ hybridization results indicated that the P-accumulating organisms (PAOs) abundance was 75 1.1% in [Formula: see text] , approximately 13.6% higher than that in a parallel P removal SBR using nitrate [Formula: see text] . Type II Accumulibacter (PAOII) (unable to use nitrate as an electron acceptor) was the main PAOs species in [Formula: see text] , contributing 72% to total PAOs. Compared with [Formula: see text] , [Formula: see text] biomass had enhanced nitrite/free nitrous acid (FNA) endurance, as demonstrated by its higher nitrite denitrification and P uptake rates. N2O accumulated temporarily in [Formula: see text] after each pulse of nitrite. Peak N2O concentrations in the bulk for [Formula: see text] were generally 6-11 times higher than that in [Formula: see text] ; these accumulations were rapidly denitrified to nitrogen gases. N2O concentration increased rapidly in nitrate-cultivated biomass when 5 or 10 mg NO2(-)-N/L per pulse was added. Whereas, N2O accumulation did not occur in nitrite-cultivated biomass until up to 30 mg NO2(-)-N/L per pulse was added. Long-term acclimation to nitrite and pulse addition of nitrite in [Formula: see text] reduced the risk of nitrite accumulation, and mitigated N2O accumulation and emissions from denitrifying P removal by nitrite. PMID:25261626

  15. Classification of positive blood cultures

    Gradel, Kim O; Knudsen, Inge Jenny Dahl; Arpi, Rolf Magnus; Ostergaard, Christian; Schnheyder, Henrik Carl; Sgaard, Mette

    2012-01-01

    ABSTRACT: BACKGROUND: Information from blood cultures is utilized for infection control, public health surveillance, and clinical outcome research. This information can be enriched by physicians assessments of positive blood cultures, which are, however, often available from selected patient groups...... or pathogens only. The aim of this work was to determine whether patients with positive blood cultures can be classified effectively for outcome research in epidemiological studies by the use of administrative data and computer algorithms, taking physicians assessments as reference. METHODS......: Physicians assessments of positive blood cultures were routinely recorded at two Danish hospitals from 2006 through 2008. The physicians assessments classified positive blood cultures as: a) contamination or bloodstream infection; b) bloodstream infection as mono- or polymicrobial; c) bloodstream infection...

  16. Uranium enrichment and the environment

    The three gaseous diffusion plants comprise one of the largest industrial complexes in the free world. Since all of the enrichment is done inside a very large plumbing maze, the control of radioactivity does not present a serious problem. Conventional, nonexotic engineering and administrative measures adequately control the minor levels of radioactivity associated with support activities such as equipment decontamination and maintenance. The treatment and control of chemical waste streams to comply with Federal and state regulations has required the commitment of 47 million dollars since 1974. Through 1982, an additional 84 million dollars may be required. The high cost is not a reflection of initially poor conditions but is rather the result of providing large treatment systems to meet very low discharge limits. Examples that will be discussed include airborne particulate removal, recirculating cooling water treatment, and hazardous waste disposal concepts

  17. Enriched natural zeolites ? mineral fertilizer

    Rybrov Lucia

    2001-12-01

    Full Text Available Clinoptilolit belongs to the big group of minerals called zeolites. Chemically they are alumosilicates and have a very difficult crystal structure. Nature zeolites are used as sorbents, molecular sieves and catalyzators. Their application is also in agriculture and horticulture. Endeavour of all agricurtural subjects from the plant area is to reach a maximum efficiency and production.Charges for the fertilization are 14 ? 40% of all charges depending on the structure of product, intensity of production and share of nutrition in soil. For the elimination of the loss in the nutrition matter, it is possible to use a ?porter?, which secures a regular and inteligent supply of nutrition to plants.Regarding the physical and chemical properties is good to use natural zeolites as a ?porter?.The experiments were made with enriched zeolites, and their influences an vegetables and flowers roots was reserched. Results were positive.

  18. Gasket for uranium enrichment plant

    Purpose: To obtain a gasket to be inserted between flange joints in the equipments and pipe lines of an uranium enrichment plant having neither permeability nor adsorptivity to water while maintaining mechanical, physical and chemical properties of an elastomer gasket. Constitution: A gasket made of an elastomeric material such as a polymer is integratedly formed at its surface with anti-slip projections. The gasket is further surrounded at its upper and lower peripheral sides, as well as outer circumferential portion with an U-sectioned cover (enclosure) made of fluoro-plastics. In this arrangement, the gasket main body shows a gas-tightness for uranium hexafluoride gas and the cover exhibits a gas-tightness for other component gases such as moisture to thereby prevent degradation of the gasket due to absorption and permeation of the moisture. (Furukawa, Y.)

  19. Stable isotope enrichment: Current and future potential

    Oak Ridge National Laboratory (ORNL) operates the Isotope Enrichment Facility for the purpose of providing enriched stable isotopes, selected radioactive isotopes (including the actinides), and isotope-related materials and services for use in various research applications. ORNL is responsible for isotope enrichment and the distribution of approximately 225 nongaseous stable isotopes from 50 multi-isotopic elements. Many enriched isotope products are of prime importance in the fabrication of nuclear targets and the subsequent production of special radionuclides. State-of-the-art techniques to achieve special isotopic, chemical, and physical requirements are performed at ORNL This report describes the status and capabilities of the Isotope Enrichment Facility and the Isotope Research Materials Laboratory as well as emphasizing potential advancements in enrichment capabilities

  20. Experimental comparison of uranium enrichment measurement methods

    Park, Se-Hwan; Eom, Sung-Ho; Shin, Hee-Sung; Kim, Ho Dong; Park, June-Sic

    2012-08-01

    Uranium enrichment (the weight fraction of the fissile material, 235U, in the uranium sample) can be determined with passive or active methods. In the present work, the characteristics of uranium enrichment measurement methods were measured and compared. Especially, the applicability of a low-intensity neutron generator to the uranium enrichment measurement was investigated. The γ- ray energy spectra from uranium samples (enrichment from 1.5% to 4.5%) were measured passively. The delayed γ-ray spectra from the uranium were measured by interrogating with a continuous neutron beam or a pulsed neutron beam. The relation between the γ-ray count rate and the uranium enrichment was obtained for each measurement. The background contribution to the total count rate could be minimized and the sensitivity of the count rate to the uranium enrichment was larger when the γ-ray spectrum was measured with a pulsed neutron beam.