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Sample records for acidophilic methanotroph isolate

  1. Complete genome sequence of the extremely acidophilic methanotroph isolate V4, Methylacidiphilum infernorum, a representative of the bacterial phylum Verrucomicrobia

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    Stott Matthew B

    2008-07-01

    Full Text Available Abstract Background The phylum Verrucomicrobia is a widespread but poorly characterized bacterial clade. Although cultivation-independent approaches detect representatives of this phylum in a wide range of environments, including soils, seawater, hot springs and human gastrointestinal tract, only few have been isolated in pure culture. We have recently reported cultivation and initial characterization of an extremely acidophilic methanotrophic member of the Verrucomicrobia, strain V4, isolated from the Hell's Gate geothermal area in New Zealand. Similar organisms were independently isolated from geothermal systems in Italy and Russia. Results We report the complete genome sequence of strain V4, the first one from a representative of the Verrucomicrobia. Isolate V4, initially named "Methylokorus infernorum" (and recently renamed Methylacidiphilum infernorum is an autotrophic bacterium with a streamlined genome of ~2.3 Mbp that encodes simple signal transduction pathways and has a limited potential for regulation of gene expression. Central metabolism of M. infernorum was reconstructed almost completely and revealed highly interconnected pathways of autotrophic central metabolism and modifications of C1-utilization pathways compared to other known methylotrophs. The M. infernorum genome does not encode tubulin, which was previously discovered in bacteria of the genus Prosthecobacter, or close homologs of any other signature eukaryotic proteins. Phylogenetic analysis of ribosomal proteins and RNA polymerase subunits unequivocally supports grouping Planctomycetes, Verrucomicrobia and Chlamydiae into a single clade, the PVC superphylum, despite dramatically different gene content in members of these three groups. Comparative-genomic analysis suggests that evolution of the M. infernorum lineage involved extensive horizontal gene exchange with a variety of bacteria. The genome of M. infernorum shows apparent adaptations for existence under extremely

  2. Acidophilic algae isolated from mine-impacted environments and their roles in sustaining heterotrophic acidophiles

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    David Barrie Johnson

    2012-09-01

    Full Text Available Two acidophilic algae, identified as strains of Chlorella protothecoides var. acidicola and Euglena mutabilis, were isolated in pure culture from abandoned copper mines in Spain and Wales and grown in pH- and temperature-controlled bioreactors. The Chlorella isolate grew optimally at pH 2.5 and 30 ˚C, with a corresponding culture doubling time of 9 hours. The isolates displayed similar tolerance (10-50 mM to four transition metals tested. Growth of the algae in liquid media was paralleled with increasing concentrations of dissolved organic carbon (DOC. Glycolic acid was identified as a significant component (12- 14% of total DOC. Protracted incubation resulted in concentrations of glycolic acid declining in both cases, and glycolic acid added to a culture of Chlorella incubated in the dark was taken up by the alga (~100% within three days. Two monosaccharides were identified in cell-free liquors of each algal isolate: fructose and glucose (Chlorella, and mannitol and glucose (Euglena. These were rapidly metabolised by acidophilic heterotrophic bacteria (Acidiphilium and Acidobacterium spp. though only fructose was utilised by the more fastidious heterotroph Acidocella aromatica. The significance of algae in promoting the growth of iron- (and sulfate- reducing heterotrophic acidophiles that are important in remediating mine-impacted waters is discussed.

  3. Role of Acidophilic Methanotrophs in Long Term Natural Attenuation of cVOCs in Low pH Aquifers

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    2017-06-01

    bacteria in acidic peatlands using newly developed 16S rRNA-targeted fluorescent oligonucleotide probes. FEMS Microbiol. Ecol. 43:299-308. Dedysh S.N...Methylocystis strain SB2 grown on ethanol: bioremediation via facultative methanotrophy. FEMS Microbiol. Lett. 318:137–142. Islam T., Torsvik V., Larsen Ø...Verrumicrobia. Appl. Environ. Microbiol. 71:8402-8410. Semrau J.D., DiSpirito A.A., Yoon, S. 2010. Methanotrophs and copper. FEMS Microbiol. Rev. 34

  4. Biological conversion of biogas to methanol using methanotrophs isolated from solid-state anaerobic digestate.

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    Sheets, Johnathon P; Ge, Xumeng; Li, Yueh-Fen; Yu, Zhongtang; Li, Yebo

    2016-02-01

    The aim of this work was to isolate methanotrophs (methane oxidizing bacteria) that can directly convert biogas produced at a commercial anaerobic digestion (AD) facility to methanol. A methanotrophic bacterium was isolated from solid-state anaerobic digestate. The isolate had characteristics comparable to obligate methanotrophs from the genus Methylocaldum. This newly isolated methanotroph grew on biogas or purified CH4 and successfully converted biogas from AD to methanol. Methanol production was achieved using several methanol dehydrogenase (MDH) inhibitors and formate as an electron donor. The isolate also produced methanol using phosphate with no electron donor or using formate with no MDH inhibitor. The maximum methanol concentration (0.43±0.00gL(-1)) and 48-h CH4 to methanol conversion (25.5±1.1%) were achieved using biogas as substrate and a growth medium containing 50mM phosphate and 80mM formate.

  5. Ardrea characterisation of acidophilic micro-organisms isolated from gold mines in Marmato, Colombia

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    Edna Judith Márquez F.

    2008-02-01

    Full Text Available Mineral bio-oxidation improves the extraction of valuable metals and also decreases the impact caused by mining waste; however, the interactions between the micro-organisms so involved are little known. Double-layer solid culture media techniques and amplified ribosomal DNA restriction enzyme analysis (Ardrea, using Eco72I, Eco24I, XcmI and BsaAI enzymes, were used for characterising four micro-organisms isolated from gold mines located in Marmato, Colombia. This work was aimed at better understanding of native acidophilic micro-organisms’ microbial interactions in mixed cultures. Iron and sulphur oxidising isolates revealed similar restriction patterns to those previously reported for Acidithiobacillus ferrooxidans; however, one of them exhibited different colony morphology compared to previously reported morphology. The iron non-oxidising isolate presented a restriction pattern agreeing with theoretical analysis of Acidithiobacillus thiooxidans database sequences. ARDREA proved to be a viable technique for differentiating between At. ferrooxidans and At. thiooxidans; in turn, it enabled checking isolates’ identity with their physiological traits and colony morphology.

  6. Isolation of methanotrophic bacteria from a london landfill: a preliminary study using molecular and stable isotopic techniques.

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    Sriskantharajah, S.; Cutting, S.; Lowry, D.; Grassineau, N.; Nisbet, E.

    2003-04-01

    Methane emissions from landfills are an important source of European greenhouse emissions, and could be reduced by a biological management program that used methanotrophs in landfill cover soils. Topsoil samples taken from a London Landfill were incubated on Nitrate Mineral Salts medium in the presence of methane. The resulting colonies were probed for methanotrophic DNA using PCR amplification. DNA from methanotroph positive colonies was cloned and sequenced for identification. Isolates belonging to the genera Methylocaldum, Methylomonas and Methylosinus were detected. Phylogenetic analysis suggests the presence of possible new species. In addition dried samples of the isolates were analysed for their stable carbon isotope (δ 13C) composition. The results were δ 13C values of -27 per mil and -25 per mil for Methylomonas isolates, -35 per mil and -44 per mil for Methylosinus isolates, -58 per mil and -60 per mil for some of the Methylocaldum isolates and -35 per mil and -45 per mil for the others. This isotopic variation is reflected in a phylogenetic tree of the isolates. The differences shown in the δ 13C analysis could be due to differing biochemical properties, and if the technique is further developed, it may be used for rapid identification of bacteria useful in landfill management for reducing methane emissions. The results suggest that useful reductions in methane emissions could be achieved by a careful design of landfill cover to culture methanotrophs.

  7. Purification and biochemical characterization of an acidophilic amylase from a newly isolated Bacillus sp. DR90.

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    Asoodeh, Ahmad; Alemi, Ashraf; Heydari, Akbar; Akbari, Jafar

    2013-03-01

    An acidophilic and Ca(2+)-independent amylase was purified from a newly isolated Bacillus sp. DR90 by ion-exchange chromatography, and exhibited a molecular weight of 68.9 kDa by SDS-PAGE. The optimum pH and temperature of the enzyme were found to be 4.0 and 45 °C, respectively. The enzyme activity was increased by Ba(2+), Fe(2+) and Mg(2+), and decreased by Hg(2+) and Zn(2+), while it was not affected by Na(+), K(+), phenylmethylsulfonyl fluoride and β-mercaptoethanol. Ca(2+) and EDTA did not have significant effect on the enzyme activity and thermal stability. The values of K m and V max for starch as substrate were 4.5 ± 0.13 mg/ml and 307 ± 12 μM/min/mg, respectively. N,N-dialkylimidazolium-based ionic liquids such as 1-hexyl-3-methylimidazolium bromide [HMIM][Br] have inhibitory effect on the enzyme activity. Thin layer chromatography analyses displayed that maltose and glucose are the main products of the enzyme reaction on starch. Regarding the features of the enzyme, it may be utilized as a novel candidate for industrial applications.

  8. Isolation and identification of moderately thermophilic acidophilic iron-oxidizing bacterium and its bioleaching characterization

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    ZENG Wei-min; WU Chang-bin; ZHANG Ru-bing; HU Pei-lei; QIU Guan-zhou; GU Guo-hua; ZHOU Hong-bo

    2009-01-01

    A moderately thermophilic acidophilic iron-oxidizing bacterium ZW-1 was isolated from Dexing mine, Jiangxi Province, China. The morphological, biochemical and physiological characteristics, 16S rRNA sequence and bioleaching characterization of strain ZW-1 were studied. The optimum growth temperature is 48 ℃, and the optimum initial pH is 1.9. The strain can grow autotrophically by using ferrous iron or elemental sulfur as sole energy sources. The strain is also able to grow heterotrophically by using peptone and yeast extract powder, but not glucose. The cell density of strain ZW-1 can reach up to 1.02×108 /mL with addition of 0.4 g/L peptone. A phylogenetic tree was constructed by comparing with the published 16S rRNA sequences of the relative bacteria species. In the phylogenetic tree, strain ZW-1 is closely relative to Sulfobacilus acidophilus with more than 99% sequence similarity. The results of bioleaching experiments indicate that the strain could oxidize Fe2+ efficiently, and the maximum oxidizing rate is 0.295 g/(L·h). It could tolerate high concentration of Fe3+ and Cu2+ (35 g/L and 25 g/L, respectively). After 20 d, 44.6% of copper is extracted from chalcopyrite by using strain ZW-1 as inocula.

  9. Methylohalobius crimeensis gen. nov., sp. nov., a moderately halophilic, methanotrophic bacterium isolated from hypersaline lakes of Crimea.

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    Heyer, Jürgen; Berger, Ursula; Hardt, Martin; Dunfield, Peter F

    2005-09-01

    A novel genus and species are proposed for two strains of methanotrophic bacteria isolated from hypersaline lakes in the Crimean Peninsula of Ukraine. Strains 10Ki(T) and 4Kr are moderate halophiles that grow optimally at 1-1.5 M (5.8-8.7%, w/v) NaCl and tolerate NaCl concentrations from 0.2 M up to 2.5 M (1.2-15%). This optimum and upper limit are the highest for any methanotrophic bacterium known to date. The strains are Gram-negative, aerobic, non-pigmented, motile, coccoid to spindle-shaped bacteria that grow on methane or methanol only and utilize the ribulose monophosphate pathway for carbon assimilation. They are neutrophilic (growth occurs only in the range pH 6.5-7.5) and mesophilic (optimum growth occurs at 30 degrees C). On the basis of 16S rRNA gene sequence phylogeny, strains 10Ki(T) and 4Kr represent a type I methanotroph within the 'Gammaproteobacteria'. However, the 16S rRNA gene sequence displays <91.5 % identity to any public-domain sequence. The most closely related methanotrophic bacterium is the thermophilic strain HB. The DNA G+C content is 58.7 mol%. The major phospholipid fatty acids are 18:1omega7 (52-61%), 16:0 (22-23%) and 16:1omega7 (14-20%). The dominance of 18:1 over 16:0 and 16:1 fatty acids is unique among known type I methanotrophs. The data suggest that strains 10Ki(T) and 4Kr should be considered as belonging to a novel genus and species of type I methanotrophic bacteria, for which the name Methylohalobius crimeensis gen. nov., sp. nov. is proposed. Strain 10Ki(T) (=DSM 16011(T)=ATCC BAA-967(T)) is the type strain.

  10. Diversity and Ecophysiology of New Isolates of Extremely Acidophilic CS2-Converting Acidithiobacillus Strains

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    Smeulders, Marjan J.; Pol, Arjan; Zandvoort, Marcel H.; Jetten, Mike S. M.

    2013-01-01

    Biofiltration of industrial carbon disulfide (CS2)-contaminated waste air streams results in the acidification of biofilters and therefore reduced performance, high water use, and increased costs. To address these issues, we isolated 16 extremely acidophilic CS2-converting Acidithiobacillus thiooxidans strains that tolerated up to 6% (vol/vol) sulfuric acid. The ecophysiological properties of five selected strains (2Bp, Sts 4-3, S1p, G8, and BBW1) were compared. These five strains had pH optima between 1 (2Bp) and 2 (S1p). Their affinities for CS2 ranged between 80 (G8) and 130 (2Bp) μM. Strains S1p, G8, and BBW1 had more hydrophobic cell surfaces and produced less extracellular polymeric substance than did strains 2Bp and Sts 4-3. All five strains converted about 80% of the S added as CS2 to S0 when CS2 was supplied in excess. The rate of S0 consumption varied between 7 (Sts 4-3) and 63 (S1p) nmol O2 min−1 ml culture−1. Low S0 consumption rates correlated partly with low levels of cell attachment to externally produced S0 globules. During chemostat growth, the relative amount of CS2 hydrolase in the cell increased with decreasing growth rates. This resulted in more S0 accumulation during CS2 overloads at low growth rates. Intermittent interruptions of the CS2 supply affected all five strains. Strains S1p, G8, and BBW1 recovered from 24 h of starvation within 4 h, and strains 2Bp and Sts 4-3 recovered within 24 h after CS2 was resupplied. We recommend the use of mixtures of Acidithiobacillus strains in industrial biofilters. PMID:23995926

  11. Draft Genome Sequence of "Acidibacillus ferrooxidans" ITV01, a Novel Acidophilic Firmicute Isolated from a Chalcopyrite Mine Drainage Site in Brazil.

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    Dall'Agnol, Hivana; Ñancucheo, Ivan; Johnson, D Barrie; Oliveira, Renato; Leite, Laura; Pylro, Victor S; Holanda, Roseanne; Grail, Barry; Carvalho, Nelson; Nunes, Gisele Lopes; Tzotzos, George; Fernandes, Gabriel Rocha; Dutra, Julliane; Orellana, Sara Cuadros; Oliveira, Guilherme

    2016-03-17

    Here, we report the draft genome sequence of "Acidibacillus ferrooxidans" strain ITV01, a ferrous iron- and sulfide-mineral-oxidizing, obligate heterotrophic, and acidophilic bacterium affiliated with the phylum Firmicutes. Strain ITV01 was isolated from neutral drainage from a low-grade chalcopyrite from a mine in northern Brazil.

  12. Draft Genome Sequence of “Acidibacillus ferrooxidans” ITV01, a Novel Acidophilic Firmicute Isolated from a Chalcopyrite Mine Drainage Site in Brazil

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    Dall’Agnol, Hivana; Ñancucheo, Ivan; Johnson, D. Barrie; Oliveira, Renato; Leite, Laura; Holanda, Roseanne; Grail, Barry; Carvalho, Nelson; Nunes, Gisele Lopes; Tzotzos, George; Fernandes, Gabriel Rocha; Dutra, Julliane; Orellana, Sara Cuadros

    2016-01-01

    Here, we report the draft genome sequence of “Acidibacillus ferrooxidans” strain ITV01, a ferrous iron- and sulfide-mineral-oxidizing, obligate heterotrophic, and acidophilic bacterium affiliated with the phylum Firmicutes. Strain ITV01 was isolated from neutral drainage from a low-grade chalcopyrite from a mine in northern Brazil. PMID:26988062

  13. Methanobactin: a copper binding compound having antibiotic and antioxidant activity isolated from methanotrophic bacteria

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    DiSpirito, Alan A.; Zahn, James A.; Graham, David W.; Kim, Hyung J.; Alterman, Michail; Larive, Cynthia

    2007-04-03

    A means and method for treating bacterial infection, providing antioxidant activity, and chelating copper using a copper binding compound produced by methanotrophic bacteria is described. The compound, known as methanobactin, is the first of a new class of antibiotics having gram-positive activity. Methanobactin has been sequenced, and its structural formula determined.

  14. Alicyclobacillus sp. strain CC2, a thermo-acidophilic bacterium isolated from Deception Island (Antarctica) containing a thermostable superoxide dismutase enzyme

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    Daniela N. Correa-Llantén; Maximiliano J. Amenábar; Patricio A. Muñoz; María T. Monsalves; Miguel E. Castro; Jenny M.Blamey

    2014-01-01

    A gram-positive, rod-shaped, aerobic, thermo-acidophilic bacterium CC2 (optimal temperature 55℃and pH 4.0), belonging to the genus Alicyclobacillus was isolated from geothermal soil collected from“Cerro Caliente”, Deception Island, Antarctica. Owing to the harsh environmental conditions found in this territory, microorganisms are exposed to conditions that trigger the generation of reactive oxygen species (ROS). They must have an effective antioxidant defense system to deal with this oxidative stress. We focused on one of the most important enzymes: superoxide dismutase, which was partially purified and characterized. This study presents the ifrst report of a thermo-acidophilic bacterium isolated from Deception Island with a thermostable superoxide dismutase (SOD).

  15. A Description of an Acidophilic, Iron Reducer, Geobacter sp. FeAm09 Isolated from Tropical Soils

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    Healy, O.; Souchek, J.; Heithoff, A.; LaMere, B.; Pan, D.; Hollis, G.; Yang, W. H.; Silver, W. L.; Weber, K. A.

    2014-12-01

    Iron (Fe) is the fourth most abundant element in the Earth's crust and plays a significant role controlling the geochemistry in soils, sediments, and aquatic systems. As part of a study to understand microbially-catalysed iron biogeochemical cycling in tropical soils, an iron reducing isolate, strain FeAm09, was obtained. Strain FeAm09 was isolated from acidic, Fe-rich soils collected from a tropical forest (Luquillo Experimental Forest, Puerto Rico). Strain FeAm09 is a rod-shaped, motile, Gram-negative bacterium. Taxonomic analysis of the near complete 16S rRNA gene sequence revealed that strain FeAm09 is 94.7% similar to Geobacter lovleyi, placing it in the genus Geobacter within the Family Geobacteraceae in the Deltaproteobacteria. Characterization of the optimal growth conditions revealed that strain FeAm09 is a moderate acidophile with an optimal growth pH of 5.0. The optimal growth temperature was 37°C. Growth of FeAm09 was coupled to the reduction of soluble Fe(III), Fe(III)-NTA, with H2, fumarate, ethanol, and various organic acids and sugars serving as the electron donor. Insoluble Fe(III), in the form of synthetic ferrihydrite, was reduced by strain FeAm09 using acetate or H2 as the electron donor. The use of H2 as an electron donor in the presence of CO2 and absence of organic carbon and assimilation of 14C-labelled CO2 into biomass indicate that strain FeAm09 is an autotrophic Fe(III)-reducing bacterium. Together, these data describe the first acidophilic, autotrophic Geobacter species. Iron reducing bacteria were previously shown to be as abundant in tropical soils as in saturated sediments (lake-bottoms) and saturated soils (wetlands) where Fe(III) reduction is more commonly recognized as a dominant mode of microbial respiration. Furthermore, Fe(III) reduction was identified as a primary driver of carbon mineralization in these tropical soils (Dubinsky et al. 2010). In addition to mineralizing organic carbon, Geobacter sp. FeAm09 is likely to also

  16. A novel acidophilic, thermophilic iron and sulfur-oxidizing archaeon isolated from a hot spring of tengchong, yunnan, China

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    Jiannan Ding

    2011-06-01

    Full Text Available A novel thermoacidophilic iron and sulfur-oxidizing archaeon, strain YN25, was isolated from an in situ enriched acid hot spring sample collected in Yunnan, China. Cells were irregular cocci, about 0.9-1.02 µm×1.0-1.31 µm in the medium containing elemental sulfur and 1.5-2.22 µm×1.8-2.54 µm in ferrous sulfate medium. The ranges of growth and pH were 50-85 (optimum 65 and pH 1.0-6.0 (optimum 1.5-2.5. The acidophile was able to grow heterotrophically on several organic substrates, including various monosaccharides, alcohols and amino acids, though the growth on single substrate required yeast extract as growth factor. Growth occurred under aerobic conditions or via anaerobic respiration using elemental sulfur as terminal electron acceptor. Results of morphology, physiology, fatty acid analysis and analysis based on 16S rRNA gene sequence indicated that the strain YN25 should be grouped in the species Acidianus manzaensis. Bioleaching experiments indicated that this strain had excellent leaching capacity, with a copper yielding ratio up to 79.16% in 24 d. The type strain YN25 was deposited in China Center for Type Culture Collection (=CCTCCZNDX0050.

  17. Desulfosporosinus acididurans sp. nov.: an acidophilic sulfate-reducing bacterium isolated from acidic sediments

    NARCIS (Netherlands)

    Sanchez Andrea, I.; Stams, A.J.M.; Hedrich, S.; Nancucheo, I.; Johnson, D.B.

    2015-01-01

    Three strains of sulfate-reducing bacteria (M1T, D, and E) were isolated from acidic sediments (White river and Tinto river) and characterized phylogenetically and physiologically. All three strains were obligately anaerobic, mesophilic, spore-forming straight rods, stained Gram-negative and

  18. Uncovering a Microbial Enigma: Isolation and Characterization of the Streamer-Generating, Iron-Oxidizing, Acidophilic Bacterium “Ferrovum myxofaciens”

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    Hallberg, Kevin B.; Hedrich, Sabrina

    2014-01-01

    A betaproteobacterium, shown by molecular techniques to have widespread global distribution in extremely acidic (pH 2 to 4) ferruginous mine waters and also to be a major component of “acid streamer” growths in mine-impacted water bodies, has proven to be recalcitrant to enrichment and isolation. A modified “overlay” solid medium was devised and used to isolate this bacterium from a number of mine water samples. The physiological and phylogenetic characteristics of a pure culture of an isolate from an abandoned copper mine (“Ferrovum myxofaciens” strain P3G) have been elucidated. “F. myxofaciens” is an extremely acidophilic, psychrotolerant obligate autotroph that appears to use only ferrous iron as an electron donor and oxygen as an electron acceptor. It appears to use the Calvin-Benson-Bassham pathway to fix CO2 and is diazotrophic. It also produces copious amounts of extracellular polymeric materials that cause cells to attach to each other (and to form small streamer-like growth in vitro) and to different solid surfaces. “F. myxofaciens” can catalyze the oxidative dissolution of pyrite and, like many other acidophiles, is tolerant of many (cationic) transition metals. “F. myxofaciens” and related clone sequences form a monophyletic group within the Betaproteobacteria distantly related to classified orders, with genera of the family Nitrosomonadaceae (lithoautotrophic, ammonium-oxidizing neutrophiles) as the closest relatives. On the basis of the phylogenetic and phenotypic differences of “F. myxofaciens” and other Betaproteobacteria, a new family, “Ferrovaceae,” and order, “Ferrovales,” within the class Betaproteobacteria are proposed. “F. myxofaciens” is the first extreme acidophile to be described in the class Betaproteobacteria. PMID:24242243

  19. Bioprospecting for acidophilic lipid-rich green microalgae isolated from abandoned mine site water bodies.

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    Eibl, Joseph K; Corcoran, Jason D; Senhorinho, Gerusa N A; Zhang, Kejian; Hosseini, Nekoo Seyed; Marsden, James; Laamanen, Corey A; Scott, John A; Ross, Gregory M

    2014-03-26

    With fossil fuel sources in limited supply, microalgae show tremendous promise as a carbon neutral source of biofuel. Current microalgae biofuel strategies typically rely on growing high-lipid producing laboratory strains of microalgae in open raceways or closed system photobioreactors. Unfortunately, these microalgae species are found to be sensitive to environmental stresses or competition by regional strains. Contamination by invasive species can diminish productivity of commercial algal processes. A potential improvement to current strategies is to identify high-lipid producing microalgae, which thrive in selected culture conditions that reduce the risk of contamination, such as low pH. Here we report the identification of a novel high-lipid producing microalgae which can tolerate low pH growth conditions. Lig 290 is a Scenedesmus spp. isolated from a low pH waterbody (pH = 4.5) in proximity to an abandoned lignite mine in Northern Ontario, Canada. Compared to a laboratory strain of Scendesmus dimorphus, Lig 290 demonstrated robust growth rates, a strong growth profile, and high lipid production. As a consequence, Lig 290 may have potential application as a robust microalgal species for use in biofuel production.

  20. A new iron-oxidizing/O2-reducing supercomplex spanning both inner and outer membranes, isolated from the extreme acidophile Acidithiobacillus ferrooxidans.

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    Castelle, Cindy; Guiral, Marianne; Malarte, Guillaume; Ledgham, Fouzia; Leroy, Gisèle; Brugna, Myriam; Giudici-Orticoni, Marie-Thérèse

    2008-09-19

    The iron respiratory chain of the acidophilic bacterium Acidithiobacillus ferrooxidans involves various metalloenzymes. Here we demonstrate that the oxygen reduction pathway from ferrous iron (named downhill pathway) is organized as a supercomplex constituted of proteins located in the outer and inner membranes as well as in the periplasm. For the first time, the outer membrane-bound cytochrome c Cyc2 was purified, and we showed that it is responsible for iron oxidation and determined that its redox potential is the highest measured to date for a cytochrome c. The organization of metalloproteins inside the supramolecular structure was specified by protein-protein interaction experiments. The isolated complex spanning the two membranes had iron oxidase as well as oxygen reductase activities, indicating functional electron transfer between the first iron electron acceptor, Cyc2, and the Cu(A) center of cytochrome c oxidase aa(3). This is the first characterization of a respirasome from an acidophilic bacterium. In Acidithiobacillus ferrooxidans,O(2) reduction from ferrous iron must be coupled to the energy-consuming reduction of NAD(+)(P) from ferrous iron (uphill pathway) required for CO(2) fixation and other anabolic processes. Besides the proteins involved in the O(2) reduction, there were additional proteins in the supercomplex, involved in uphill pathway (bc complex and cytochrome Cyc(42)), suggesting a possible physical link between these two pathways.

  1. Expanding the verrucomicrobial methanotrophic world: description of three novel species of Methylacidimicrobium gen. nov.

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    van Teeseling, Muriel C F; Pol, Arjan; Harhangi, Harry R; van der Zwart, Sietse; Jetten, Mike S M; Op den Camp, Huub J M; van Niftrik, Laura

    2014-11-01

    Methanotrophic Verrucomicrobia have been found in geothermal environments characterized by high temperatures and low pH values. However, it has recently been hypothesized that methanotrophic Verrucomicrobia could be present under a broader range of environmental conditions. Here we describe the isolation and characterization of three new species of mesophilic acidophilic verrucomicrobial methanotrophs from a volcanic soil in Italy. The three new species showed 97% to 98% 16S rRNA gene identity to each other but were related only distantly (89% to 90% on the 16S rRNA level) to the thermophilic genus Methylacidiphilum. We propose the new genus Methylacidimicrobium, including the novel species Methylacidimicrobium fagopyrum, Methylacidimicrobium tartarophylax, and Methylacidimicrobium cyclopophantes. These mesophilic Methylacidimicrobium spp. were more acid tolerant than their thermophilic relatives; the most tolerant species, M. tartarophylax, still grew at pH 0.5. The variation in growth temperature optima (35 to 44°C) and maximum growth rates (µmax; 0.013 to 0.040 h(-1)) suggested that all species were adapted to a specific niche within the geothermal environment. All three species grew autotrophically using the Calvin cycle. The cells of all species contained glycogen particles and electron-dense particles in their cytoplasm as visualized by electron microscopy. In addition, the cells of one of the species (M. fagopyrum) contained intracytoplasmic membrane stacks. The discovery of these three new species and their growth characteristics expands the known diversity of verrucomicrobial methanotrophs and shows that they are present in many more ecosystems than previously assumed.

  2. Bacteriophages of methanotrophic bacteria

    Energy Technology Data Exchange (ETDEWEB)

    Tyutikow, F.M. (All-Union Research Inst. for Genetics and Selection of Industrial Microorganisms, Moscow, USSR); Bespalova, I.A.; Rebentish, B.A.; Aleksandrushkina, N.N.; Krivisky, A.S.

    1980-10-01

    Bacteriophages of methanotrophic bacteria have been found in 16 out of 88 studied samples (underground waters, pond water, soil, gas and oil installation waters, fermentor cultural fluids, bacterial paste, and rumen of cattle) taken in different geographic zones of the Soviet Union. Altogether, 23 phage strains were isolated. By fine structure, the phages were divided into two types (with very short or long noncontractile tails); by host range and serological properties, they fell into three types. All phages had guanine- and cytosine-rich double-stranded deoxyribonucleic acid consisting of common nitrogen bases. By all of the above-mentioned properties, all phages within each of the groups were completely identical to one another, but differed from phages of other groups.

  3. Production of poly-{beta}-hydroxybutyrate (PHB) by Methylobacterium organophilum isolated from a methanotrophic consortium in a two-phase partition bioreactor

    Energy Technology Data Exchange (ETDEWEB)

    Zuniga, C., E-mail: cristal7n@gmail.com [Universidad Autonoma Metropolitana-Cuajimalpa, Departamento de Procesos y Tecnologia, Artificios 40, Col. Miguel Hidalgo, C.P. 01120, Mexico D.F. (Mexico); Morales, M., E-mail: mmorales@correo.cua.uam.mx [Universidad Autonoma Metropolitana-Cuajimalpa, Departamento de Procesos y Tecnologia, Artificios 40, Col. Miguel Hidalgo, C.P. 01120, Mexico D.F. (Mexico); Le Borgne, S., E-mail: sylvielb@correo.cua.uam.mx [Universidad Autonoma Metropolitana-Cuajimalpa, Departamento de Procesos y Tecnologia, Artificios 40, Col. Miguel Hidalgo, C.P. 01120, Mexico D.F. (Mexico); Revah, S., E-mail: srevah@correo.cua.uam.mx [Universidad Autonoma Metropolitana-Cuajimalpa, Departamento de Procesos y Tecnologia, Artificios 40, Col. Miguel Hidalgo, C.P. 01120, Mexico D.F. (Mexico)

    2011-06-15

    The biodegradation of methane, a greenhouse gas, and the accumulation of poly-{beta}-hydroxybutyrate (PHB) were studied using a methanotrophic consortium and an isolated strain thereof. The specific rates for methane consumption were 100 and 17mg{sub CH{sub 4}}g{sub x}{sup -1} h{sup -1} for the isolate and the consortium, respectively. Also the effect of including 10% (v v{sup -1}) of silicone oil in a two-phase partitioning bioreactor (TPPB) was assayed for the elimination of 1% methane in air stream. TPPB allowed a 33-45% increase of methane elimination under growing conditions. Nitrogen limitation was assayed in bioreactors to promote PHB production. Under this condition, the specific methane degradation rate remained unchanged for the consortium and decreased to 36mg{sub CH{sub 4}}g{sub x}{sup -1} h{sup -1} for the isolated strain. The accumulated PHB in the reactor was 34% and 38% (w w{sup -1}) for the consortium and the isolate, respectively. The highest productivity was obtained in the TPPB and was 1.61mg{sub PHB}g{sub x}{sup -1} h{sup -1}. The CZ-2 isolate was identified as Methylobacterium organophilum, this is the first study that reports this species as being able to grow on methane and accumulate up to 57% (w w{sup -1}) of PHB under nitrogen limitation in microcosm experiments.

  4. Genome Sequence of the Acidophilic Bacterium Acidocella sp. Strain MX-AZ02

    DEFF Research Database (Denmark)

    Servín-Garcidueñas, Luis E.; Garrett, Roger A.; Amils, Ricardo;

    2013-01-01

    Here, we report the draft genome sequence of Acidocella sp. strain MX-AZ02, an acidophilic and heterotrophic alphaproteobacterium isolated from a geothermal lake in western Mexico.......Here, we report the draft genome sequence of Acidocella sp. strain MX-AZ02, an acidophilic and heterotrophic alphaproteobacterium isolated from a geothermal lake in western Mexico....

  5. Biodegradation of polycyclic aromatic hydrocarbons by an acidophilic Stenotrophomonas maltophilia strain AJH1 isolated from a mineral mining site in Saudi Arabia.

    Science.gov (United States)

    Arulazhagan, P; Al-Shekri, K; Huda, Q; Godon, J J; Basahi, J M; Jeyakumar, D

    2017-01-01

    The present study aims at analyzing the degradation of polycyclic aromatic hydrocarbons (PAHs) at acidic conditions (pH = 2) by acidophilic Stenotrophomonas maltophilia strain AJH1 (KU664513). The strain AJH1 was obtained from an enrichment culture obtained from soil samples of mining area in the presence of PAH as sole sources of carbon and energy. Strain AJH1was able to degrade low (anthracene, phenanthrene, naphthalene, fluorene) and high (pyrene, benzo(e)pyrene and benzo(k)fluoranthene) molecular weight PAHs in acidophilic mineral salt medium at pH 2, with removal rates of up to 95% (LMW PAH) and 80% (HMW PAH), respectively. In addition, strain AJH1 treated petroleum wastewater with 89 ± 1.1% COD removal under acidic condition (pH 2) in a continuously stirred reactor. Acidophilic S. maltophilia strain AJH1, hence holds the promise as an effective degrader for biological treatment of PAHs contaminated wastewater at acidic pH.

  6. The importance of methanotrophic activity in geothermal soils of Pantelleria island (Italy)

    Science.gov (United States)

    D'Alessandro, Walter; Gagliano, Antonina Lisa; Quatrini, Paola; Parello, Francesco

    2013-04-01

    were investigated by culturing and culture-independent techniques. The diversity of proteobacterial methanotrophs was investigated by creating a clone library of the amplified methane mono-oxygenase encoding gene, pmoA. Clone sequencing indicates the presence of Gammaproteobacteria in the soils of Favara Grande. Enrichment cultures, on a mineral medium in a CH4-enriched atmosphere, led to the isolation of different strains that were identified as Methylocistis spp., which belong to the Alphaproteobacteria. The presence of Verrucomicrobia was detected by amplification of pmoA gene using newly designed primers. Soils from Favara Grande show therefore the largest spectrum of methanotrophic microorganisms until now detected in a geothermal environment. While the presence of Verrucomicrobia in geothermal soils was predictable due to their thermophilic and acidophilic character, the presence of both Alpha and Gamma proteobacteria was unexpected. Their presence is limited to the shallowest part of the soil were temperatures are lower and is probably favored by a soil pH that is not too low (pH ~5) and their contribution to biological methane oxidation at Pantelleria is significant. Understanding the ecology of methanotrophy in geothermal sites will increase our knowledge of the role of soils in methane emissions in such environments.

  7. [Culturable psychrotolerant methanotrophic bacteria in landfill cover soil].

    Science.gov (United States)

    Kallistova, A Iu; Montonen, L; Jurgens, G; Munster, U; Kevbrina, M V; Nozhevnikova, A N

    2014-01-01

    Methanotrophs closely related to psychrotolerant members of the genera Methylobacter and Methylocella were identified in cultures enriched at 10@C from landfill cover soil samples collected in the period from April to November. Mesophilic methanotrophs of the genera Methylobacter and Methylosinus were found in cultures enriched at 20 degrees C from the same cover soil samples. A thermotolerant methanotroph related to Methylocaldum gracile was identified in the culture enriched at 40 degrees C from a sample collected in May (the temperature of the cover soil was 11.5-12.5 degrees C). In addition to methanotrophs, methylobacteria of the genera Methylotenera and Methylovorus and members of the genera Verrucomicrobium, Pseudomonas, Pseudoxanthomonas, Dokdonella, Candidatus Protochlamydia, and Thiorhodospira were also identified in the enrichment cultures. A methanotroph closely related to the psychrotolerant species Methylobacter tundripaludum (98% sequence identity of 16S r-RNA genes with the type strain SV96(T)) was isolated in pure culture. The introduction of a mixture of the methanotrophic enrichments, grown at 15 degrees C, into the landfill cover soil resulted in a decrease in methane emission from the landfill surface in autumn (October, November). The inoculum used was demonstrated to contain methanotrophs closely related to Methylobacter tundripaludum SV96.

  8. Methanotrophic activity and diversity of methanotrophs in volcanic geothermal soils at Pantelleria (Italy)

    Science.gov (United States)

    Gagliano, A. L.; D'Alessandro, W.; Tagliavia, M.; Parello, F.; Quatrini, P.

    2014-10-01

    Volcanic and geothermal systems emit endogenous gases by widespread degassing from soils, including CH4, a greenhouse gas twenty-five times as potent as CO2. Recently, it has been demonstrated that volcanic or geothermal soils are not only a source of methane, but are also sites of methanotrophic activity. Methanotrophs are able to consume 10-40 Tg of CH4 a-1 and to trap more than 50% of the methane degassing through the soils. We report on methane microbial oxidation in the geothermally most active site of Pantelleria (Italy), Favara Grande, whose total methane emission was previously estimated at about 2.5 Mg a-1 (t a-1). Laboratory incubation experiments with three top-soil samples from Favara Grande indicated methane consumption values of up to 59.2 nmol g-1 soil d.w. h-1. One of the three sites, FAV2, where the highest oxidation rate was detected, was further analysed on a vertical soil profile, the maximum methane consumption was measured in the top-soil layer, and values greater than 6.23 nmol g-1 h-1 were still detected up to a depth of 13 cm. The highest consumption rate was measured at 37 °C, but a still detectable consumption at 80 °C (> 1.25 nmol g-1 h-1) was recorded. The soil total DNA extracted from the three samples was probed by Polymerase Chain Reaction (PCR) using standard proteobacterial primers and newly designed verrucomicrobial primers, targeting the unique methane monooxygenase gene pmoA; the presence of methanotrophs was detected at sites FAV2 and FAV3, but not at FAV1, where harsher chemical-physical conditions and negligible methane oxidation were detected. The pmoA gene libraries from the most active site (FAV2) pointed to a high diversity of gammaproteobacterial methanotrophs, distantly related to Methylocaldum-Metylococcus genera, and the presence of the newly discovered acido-thermophilic Verrucomicrobia methanotrophs. Alphaproteobacteria of the genus Methylocystis were isolated from enrichment cultures under a methane

  9. Members of the methanotrophic genus Methylomarinum inhabit inland mud pots

    Directory of Open Access Journals (Sweden)

    Danielle T. Fradet

    2016-07-01

    Full Text Available Proteobacteria capable of converting the greenhouse gas methane to biomass, energy, and carbon dioxide represent a small but important sink in global methane inventories. Currently, 23 genera of methane oxidizing (methanotrophic proteobacteria have been described, although many are represented by only a single validly described species. Here we describe a new methanotrophic isolate that shares phenotypic characteristics and phylogenetic relatedness with the marine methanotroph Methylomarinum vadi. However, the new isolate derives from a terrestrial saline mud pot at the northern terminus of the Eastern Pacific Rise (EPR. This new cultivar expands our knowledge of the ecology of Methylomarinum, ultimately towards a fuller understanding of the role of this genus in global methane cycling.

  10. Soluble Methane Monooxygenase Production and Trichloroethylene Degradation by a Type I Methanotroph, Methylomonas methanica 68-1

    OpenAIRE

    Koh, Sung-Cheol; Bowman, John P.; Sayler, Gary S.

    1993-01-01

    A methanotroph (strain 68-1), originally isolated from a trichloroethylene (TCE)-contaminated aquifer, was identified as the type I methanotroph Methylomonas methanica on the basis of intracytoplasmic membrane ultrastructure, phospholipid fatty acid profile, and 16S rRNA signature probe hybridization. Strain 68-1 was found to oxidize naphthalene and TCE via a soluble methane monooxygenase (sMMO) and thus becomes the first type I methanotroph known to be able to produce this enzyme. The specif...

  11. Methane Oxidation and Molecular Characterization of Methanotrophs from a Former Mercury Mine Impoundment

    Directory of Open Access Journals (Sweden)

    Shaun M. Baesman

    2015-06-01

    Full Text Available The Herman Pit, once a mercury mine, is an impoundment located in an active geothermal area. Its acidic waters are permeated by hundreds of gas seeps. One seep was sampled and found to be composed of mostly CO2 with some CH4 present. The δ13CH4 value suggested a complex origin for the methane: i.e., a thermogenic component plus a biological methanogenic portion. The relatively 12C-enriched CO2 suggested a reworking of the ebullitive methane by methanotrophic bacteria. Therefore, we tested bottom sediments for their ability to consume methane by conducting aerobic incubations of slurried materials. Methane was removed from the headspace of live slurries, and subsequent additions of methane resulted in faster removal rates. This activity could be transferred to an artificial, acidic medium, indicating the presence of acidophilic or acid-tolerant methanotrophs, the latter reinforced by the observation of maximum activity at pH = 4.5 with incubated slurries. A successful extraction of sterol and hopanoid lipids characteristic of methanotrophs was achieved, and their abundances greatly increased with increased sediment methane consumption. DNA extracted from methane-oxidizing enrichment cultures was amplified and sequenced for pmoA genes that aligned with methanotrophic members of the Gammaproteobacteria. An enrichment culture was established that grew in an acidic (pH 4.5 medium via methane oxidation.

  12. Methane Oxidation and Molecular Characterization of Methanotrophs from a Former Mercury Mine Impoundment.

    Science.gov (United States)

    Baesman, Shaun M; Miller, Laurence G; Wei, Jeremy H; Cho, Yirang; Matys, Emily D; Summons, Roger E; Welander, Paula V; Oremland, Ronald S

    2015-06-23

    The Herman Pit, once a mercury mine, is an impoundment located in an active geothermal area. Its acidic waters are permeated by hundreds of gas seeps. One seep was sampled and found to be composed of mostly CO₂ with some CH₄ present. The δ(13)CH₄ value suggested a complex origin for the methane: i.e., a thermogenic component plus a biological methanogenic portion. The relatively (12)C-enriched CO₂ suggested a reworking of the ebullitive methane by methanotrophic bacteria. Therefore, we tested bottom sediments for their ability to consume methane by conducting aerobic incubations of slurried materials. Methane was removed from the headspace of live slurries, and subsequent additions of methane resulted in faster removal rates. This activity could be transferred to an artificial, acidic medium, indicating the presence of acidophilic or acid-tolerant methanotrophs, the latter reinforced by the observation of maximum activity at pH = 4.5 with incubated slurries. A successful extraction of sterol and hopanoid lipids characteristic of methanotrophs was achieved, and their abundances greatly increased with increased sediment methane consumption. DNA extracted from methane-oxidizing enrichment cultures was amplified and sequenced for pmoA genes that aligned with methanotrophic members of the Gammaproteobacteria. An enrichment culture was established that grew in an acidic (pH 4.5) medium via methane oxidation.

  13. Methane oxidation and molecular characterization of methanotrophs from a former mercury mine impoundment

    Science.gov (United States)

    Baesman, Shaun; Miller, Laurence G.; Wei, Jeremy H.; Cho, Yirang; Matys, Emily D.; Summons, Roger E.; Welander, Paula V.; Oremland, Ronald S.

    2015-01-01

    The Herman Pit, once a mercury mine, is an impoundment located in an active geothermal area. Its acidic waters are permeated by hundreds of gas seeps. One seep was sampled and found to be composed of mostly CO2 with some CH4 present. The δ13CH4 value suggested a complex origin for the methane: i.e., a thermogenic component plus a biological methanogenic portion. The relatively 12C-enriched CO2 suggested a reworking of the ebullitive methane by methanotrophic bacteria. Therefore, we tested bottom sediments for their ability to consume methane by conducting aerobic incubations of slurried materials. Methane was removed from the headspace of live slurries, and subsequent additions of methane resulted in faster removal rates. This activity could be transferred to an artificial, acidic medium, indicating the presence of acidophilic or acid-tolerant methanotrophs, the latter reinforced by the observation of maximum activity at pH = 4.5 with incubated slurries. A successful extraction of sterol and hopanoid lipids characteristic of methanotrophs was achieved, and their abundances greatly increased with increased sediment methane consumption. DNA extracted from methane-oxidizing enrichment cultures was amplified and sequenced for pmoA genes that aligned with methanotrophic members of the Gammaproteobacteria. An enrichment culture was established that grew in an acidic (pH 4.5) medium via methane oxidation.

  14. Selective grazing of methanotrophs by protozoa in a rice field soil.

    Science.gov (United States)

    Murase, Jun; Frenzel, Peter

    2008-09-01

    Biological methane oxidation is a key process in the methane cycle of wetland ecosystems. The methanotrophic biomass may be grazed by protozoa, thus linking the methane cycle to the soil microbial food web. In the present study, the edibility of different methanotrophs for soil protozoa was compared. The number of methanotroph-feeding protozoa in a rice field soil was estimated by determining the most-probable number (MPN) using methanotrophs as food bacteria; naked amoebae and flagellates were the dominant protozoa. Among ten methanotrophic strains examined as a food source, seven yielded a number of protozoa comparable with the yield with Escherichia coli [10(4) MPN (g soil dry weight)(-1)], and three out of four Methylocystis spp. yielded significantly fewer numbers [10(2)-10(3) MPN (g soil dry weight)(-1)]. The lower edibility of the Methylocystis spp. was not explained either by their growth phase or by harmful effects on protozoa. Incubation of the soil under methane resulted in a higher number of protozoa actively grazing on methanotrophs, especially on the less-edible group. Protozoa isolated from the soil demonstrated a grazing preference on the different methanotrophs consistent with the results of MPN counts. The results indicate that selective grazing by protozoa may be a biological factor affecting the methanotrophic community in a wetland soil.

  15. The methanol dehydrogenase gene, mxaF, as a functional and phylogenetic marker for proteobacterial methanotrophs in natural environments.

    Directory of Open Access Journals (Sweden)

    Evan Lau

    Full Text Available The mxaF gene, coding for the large (α subunit of methanol dehydrogenase, is highly conserved among distantly related methylotrophic species in the Alpha-, Beta- and Gammaproteobacteria. It is ubiquitous in methanotrophs, in contrast to other methanotroph-specific genes such as the pmoA and mmoX genes, which are absent in some methanotrophic proteobacterial genera. This study examined the potential for using the mxaF gene as a functional and phylogenetic marker for methanotrophs. mxaF and 16S rRNA gene phylogenies were constructed based on over 100 database sequences of known proteobacterial methanotrophs and other methylotrophs to assess their evolutionary histories. Topology tests revealed that mxaF and 16S rDNA genes of methanotrophs do not show congruent evolutionary histories, with incongruencies in methanotrophic taxa in the Methylococcaceae, Methylocystaceae, and Beijerinckiacea. However, known methanotrophs generally formed coherent clades based on mxaF gene sequences, allowing for phylogenetic discrimination of major taxa. This feature highlights the mxaF gene's usefulness as a biomarker in studying the molecular diversity of proteobacterial methanotrophs in nature. To verify this, PCR-directed assays targeting this gene were used to detect novel methanotrophs from diverse environments including soil, peatland, hydrothermal vent mussel tissues, and methanotroph isolates. The placement of the majority of environmental mxaF gene sequences in distinct methanotroph-specific clades (Methylocystaceae and Methylococcaceae detected in this study supports the use of mxaF as a biomarker for methanotrophic proteobacteria.

  16. Diversity of acidophilic prokaryotes at two acid mine drainage sites in Turkey.

    Science.gov (United States)

    Aytar, Pınar; Kay, Catherine Melanie; Mutlu, Mehmet Burçin; Çabuk, Ahmet; Johnson, David Barrie

    2015-04-01

    The biodiversity of acidophilic prokaryotes in two acidic (pH 2.8-3.05) mine drainage (AMD) sites (Balya and Çan) in Turkey was examined using a combined cultivation-based and cultivation-independent approach. The latter included analyzing microbial diversity using fluorescent in situ hybridization (FISH), terminal restriction enzyme fragment length polymorphism (`T-RFLP), and quantitative PCR (qPCR). Numbers of cultivatable heterotrophic acidophilic bacteria were over an order of magnitude greater than those of chemolithotrophic acidophiles in both AMD ponds examined. Isolates identified as strains of Acidithiobacillus ferrivorans, Acidiphilium organovorum, and Ferrimicrobium acidiphilum were isolated from the Balya AMD pond, and others identified as strains of Leptospirillum ferriphilum, Acidicapsa ligni, and Acidiphilium rubrum from Çan AMD. Other isolates were too distantly related (from analysis of their 16S rRNA genes) to be identified at the species level. Archaeal diversity in the two ponds appeared to be far more limited. T-RFLP and qPCR confirmed the presence of Ferroplasma-like prokaryotes, but no archaea were isolated from the two sites. qPCR generated semiquantitative data for genera of some of the iron-oxidizing acidophiles isolated and/or detected, suggesting the order of abundance was Leptospirillum > Ferroplasma > Acidithiobacillus (Balya AMD) and Ferroplasma > Leptospirillum > Acidithiobacillus (Çan AMD).

  17. The active methanotrophic community in a wetland from the High Arctic.

    Science.gov (United States)

    Graef, Christiane; Hestnes, Anne Grethe; Svenning, Mette Marianne; Frenzel, Peter

    2011-08-01

    The dominant terminal process of carbon mineralization in most freshwater wetlands is methanogenesis. With methane being an important greenhouse gas, the predicted warming of the Arctic may provide a positive feedback. However, the amount of methane released to the atmosphere may be controlled by the activity of methane-oxidizing bacteria (methanotrophs) living in the oxic surface layer of wetlands. Previously, methanotrophs have been isolated and identified by genetic profiling in High Arctic wetlands showing the presence of only a few genotypes. Two isolates from Solvatnet (Ny-Ålesund, Spitsbergen; 79°N) are available: Methylobacter tundripaludum (type I) and Methylocystis rosea (type II), raising the question whether the low diversity is a cultivation effect. We have revisited Solvatnet applying stable isotope probing (SIP) with (13) C-labelled methane. 16S rRNA profiling revealed active type I methanotrophs including M. tundripaludum, while no active type II methanotrophs were identified. These results indicate that the extant M. tundripaludum is an active methane oxidizer at its locus typicus; furthermore, Methylobacter seems to be the dominant active genus. Diversity of methanotrophs was low as compared, e.g. to wetland rice fields in the Mediterranean. This low diversity suggests a high vulnerability of Arctic methanotroph communities, which deserves more attention.

  18. [Effect of operational modes on community structure of type I methanotroph in the cover soil of municipal solid waste landfill].

    Science.gov (United States)

    Yu, Ting; He, Pin-Jing; Lü, Fan; Shao, Li-Ming

    2008-10-01

    Type I methanotroph is crucial for methane oxidization and it responses fast to the changes in environment. In this study, 16S rDNA-based denaturing gradient gel electrophoresis (DGGE) gene fingerprint technology was applied to investigate the effect of operational modes, i. e. high-density polyethylene liner (HDPE) isolation or subsurface irrigation of landfill leachate and vegetation, on community structure and diversity of type I methanotroph in soils covering municipal solid waste landfill. 16S rDNA based phylogenetic analysis reveals type I methanotroph in all tested soils belongs to Methylobacter. According to Shannon-Wiener diversity index and principal component analysis, landfill leachate subsurface irrigation and vegetation have more impact on type I methanotroph community structure and diversity than HDPE liner isolation does, and they reduce type I methanotroph diversity. Leachate irrigation is supposed to inhibit the growth of Methylobacter population. Community structure of type I methanotroph in landfill cover soil isolated by HDPE, i.e. invaded by landfill gas, shifts during long-term gas interference. When cover age is 1.5 years old, Shannon-Wiener diversity index of type I methanotroph reaches its maximum.

  19. Deciphering Community Structure of Methanotrophs Dwelling in Rice Rhizospheres of an Indian Rice Field Using Cultivation and Cultivation-Independent Approaches.

    Science.gov (United States)

    Pandit, Pranitha S; Rahalkar, Monali C; Dhakephalkar, Prashant K; Ranade, Dilip R; Pore, Soham; Arora, Preeti; Kapse, Neelam

    2016-04-01

    Methanotrophs play a crucial role in filtering out methane from habitats, such as flooded rice fields. India has the largest area under rice cultivation in the world; however, to the best of our knowledge, methanotrophs have not been isolated and characterized from Indian rice fields. A cultivation strategy composing of a modified medium, longer incubation time, and serial dilutions in microtiter plates was used to cultivate methanotrophs from a rice rhizosphere sample from a flooded rice field in Western India. We compared the cultured members with the uncultured community as revealed by three culture-independent methods. A novel type Ia methanotroph (Sn10-6), at the rank of a genus, and a putative novel species of a type II methanotroph (Sn-Cys) were cultivated from the terminal positive dilution (10(-6)). From lower dilution (10(-4)), a strain of Methylomonas spp. was cultivated. All the three culture-independent analyses, i.e., pmoA clone library, terminal restriction fragment length polymorphism (T-RFLP), and metagenomics approach, revealed the dominance of type I methanotrophs. Only metagenomic analysis showed significant presence of type II methanotrophs, albeit in lower proportion (37 %). All the three isolates showed relevance to the methanotrophic community as depicted by uncultured methods; however, the cultivated members might not be the most dominant ones. In conclusion, a combined cultivation and cultivation-independent strategy yielded us a broader picture of the methanotrophic community from rice rhizospheres of a flooded rice field in India.

  20. Distribution and diversity of Verrucomicrobia methanotrophs in geothermal and acidic environments.

    Science.gov (United States)

    Sharp, Christine E; Smirnova, Angela V; Graham, Jaime M; Stott, Matthew B; Khadka, Roshan; Moore, Tim R; Grasby, Stephen E; Strack, Maria; Dunfield, Peter F

    2014-06-01

    Recently, methanotrophic members of the phylum Verrucomicrobia have been described, but little is known about their distribution in nature. We surveyed methanotrophic bacteria in geothermal springs and acidic wetlands via pyrosequencing of 16S rRNA gene amplicons. Putative methanotrophic Verrucomicrobia were found in samples covering a broad temperature range (22.5-81.6°C), but only in acidic conditions (pH 1.8-5.0) and only in geothermal environments, not in acidic bogs or fens. Phylogenetically, three 16S rRNA gene sequence clusters of putative methanotrophic Verrucomicrobia were observed. Those detected in high-temperature geothermal samples (44.1-81.6°C) grouped with known thermoacidiphilic 'Methylacidiphilum' isolates. A second group dominated in moderate-temperature geothermal samples (22.5-40.1°C) and a representative mesophilic methanotroph from this group was isolated (strain LP2A). Genome sequencing verified that strain LP2A possessed particulate methane monooxygenase, but its 16S rRNA gene sequence identity to 'Methylacidiphilum infernorum' strain V4 was only 90.6%. A third group clustered distantly with known methanotrophic Verrucomicrobia. Using pmoA-gene targeted quantitative polymerase chain reaction, two geothermal soil profiles showed a dominance of LP2A-like pmoA sequences in the cooler surface layers and 'Methylacidiphilum'-like pmoA sequences in deeper, hotter layers. Based on these results, there appears to be a thermophilic group and a mesophilic group of methanotrophic Verrucomicrobia. However, both were detected only in acidic geothermal environments.

  1. Methanotrophs Contribute to Peatland Nitrogen

    Science.gov (United States)

    Larmola, Tuula; Leppänen, Sanna M.; Tuittila, Eeva-Stiina; Aarva, Maija; Merilä, Päivi; Fritze, Hannu; Tiirola, Marja

    2016-04-01

    Atmospheric nitrogen (N2) fixation is potentially an important N input mechanism to peatland ecosystems, but the extent of this process may have been underestimated because of the methods traditionally used inhibit the activity of methanothrophs. We examined the linkage of methane (CH4) oxidation and N2 fixation using 15N2 technique. Dominant flark and hummock Sphagnum species were collected from twelve pristine peatlands in Siikajoki, Finland, which varied in age from 200 to 2,500 y due to the postglacial rebound. The mosses were incubated in a two-day field 15N2 and 13CH4 pulse labelling experiment and the incorporation of 15N2 and 13CH4 in biomass was measured with Isotope Ratio Mass Spectrometer. The rates of Sphagnum-associated N2 fixation (0.1-2.9 g N m-2 y-1) were up to 10 times the current N deposition rates. Methane-induced N2 fixation contributed to over 1/3 of moss-associated N2 fixation in younger stages, but was switched off in old successional stages, despite active CH4 oxidation in these stages. Both the N2 fixation rates and the methanotrophic contribution to N2 fixation during peatland succession were primarily constrained by phosphorus availability. Previously overlooked methanotrophic N contribution may explain rapid peat and N accumulation during fen stages of peatland development. Reference. Larmola T., Leppänen S.M., Tuittila E.-S, Aarva M., Merilä P., Fritze H., Tiirola M. (2014) Methanotrophy induces nitrogen fixation during peatland development. Proceedings of the National Academy of Sciences USA 111 (2): 734-739.

  2. Bioleaching of metals from electronic scrap by moderately thermophilic acidophilic bacteria

    NARCIS (Netherlands)

    Ilyas, Sadia; Anwar, Munir A.; Niazi, Shahida B.; Ghauri, M. Afzal

    2007-01-01

    The present work was aimed at studying the bioleachability of metals from electronic scrap by the selected moderately thermophilic strains of acidophilic chemolithotrophic and acidophilic heterotrophic bacteria. These included Sulfobacillus thermosulfidooxidans and an unidentified acidophilic hetero

  3. Genomics and Metagenomics of Extreme Acidophiles in Biomining Environments

    Science.gov (United States)

    Holmes, D. S.

    2015-12-01

    Over 160 draft or complete genomes of extreme acidophiles (pH models of the ecophysiology of biomining environments and provide insight into the gene and genome evolution of extreme acidophiles. Additionally, since most of these acidophiles are also chemoautolithotrophs that use minerals as energy sources or electron sinks, their genomes can be plundered for clues about the evolution of cellular metabolism and bioenergetic pathways during the Archaean abiotic/biotic transition on early Earth. Acknowledgements: Fondecyt 1130683.

  4. Enhanced Productivity of a Lutein-Enriched Novel Acidophile Microalga Grown on Urea

    Science.gov (United States)

    Casal, Carlos; Cuaresma, Maria; Vega, Jose Maria; Vilchez, Carlos

    2011-01-01

    Coccomyxa acidophila is an extremophile eukaryotic microalga isolated from the Tinto River mining area in Huelva, Spain. Coccomyxa acidophila accumulates relevant amounts of β-carotene and lutein, well-known carotenoids with many biotechnological applications, especially in food and health-related industries. The acidic culture medium (pH < 2.5) that prevents outdoor cultivation from non-desired microorganism growth is one of the main advantages of acidophile microalgae production. Conversely, acidophile microalgae growth rates are usually very low compared to common microalgae growth rates. In this work, we show that mixotrophic cultivation on urea efficiently enhances growth and productivity of an acidophile microalga up to typical values for common microalgae, therefore approaching acidophile algal production towards suitable conditions for feasible outdoor production. Algal productivity and potential for carotenoid accumulation were analyzed as a function of the nitrogen source supplied. Several nitrogen conditions were assayed: nitrogen starvation, nitrate and/or nitrite, ammonia and urea. Among them, urea clearly led to the best cell growth (~4 × 108 cells/mL at the end of log phase). Ammonium led to the maximum chlorophyll and carotenoid content per volume unit (220 μg·mL·1 and 35 μg·mL·1, respectively). Interestingly, no significant differences in growth rates were found in cultures grown on urea as C and N source, with respect to those cultures grown on nitrate and CO2 as nitrogen and carbon sources (control cultures). Lutein accumulated up to 3.55 mg·g·1 in the mixotrophic cultures grown on urea. In addition, algal growth in a shaded culture revealed the first evidence for an active xanthophylls cycle operative in acidophile microalgae. PMID:21339944

  5. Enhanced Productivity of a Lutein-Enriched Novel Acidophile Microalga Grown on Urea

    Directory of Open Access Journals (Sweden)

    Carlos Vilchez

    2010-12-01

    Full Text Available Coccomyxa acidophila is an extremophile eukaryotic microalga isolated from the Tinto River mining area in Huelva, Spain. Coccomyxa acidophila accumulates relevant amounts of b-carotene and lutein, well-known carotenoids with many biotechnological applications, especially in food and health-related industries. The acidic culture medium (pH < 2.5 that prevents outdoor cultivation from non-desired microorganism growth is one of the main advantages of acidophile microalgae production. Conversely, acidophile microalgae growth rates are usually very low compared to common microalgae growth rates. In this work, we show that mixotrophic cultivation on urea efficiently enhances growth and productivity of an acidophile microalga up to typical values for common microalgae, therefore approaching acidophile algal production towards suitable conditions for feasible outdoor production. Algal productivity and potential for carotenoid accumulation were analyzed as a function of the nitrogen source supplied. Several nitrogen conditions were assayed: nitrogen starvation, nitrate and/or nitrite, ammonia and urea. Among them, urea clearly led to the best cell growth (~4 ´ 108 cells/mL at the end of log phase. Ammonium led to the maximum chlorophyll and carotenoid content per volume unit (220 mg·mL-1 and 35 mg·mL-1, respectively. Interestingly, no significant differences in growth rates were found in cultures grown on urea as C and N source, with respect to those cultures grown on nitrate and CO2 as nitrogen and carbon sources (control cultures. Lutein accumulated up to 3.55 mg·g-1 in the mixotrophic cultures grown on urea. In addition, algal growth in a shaded culture revealed the first evidence for an active xanthophylls cycle operative in acidophile microalgae.

  6. Genome Sequence of Desulfosporosinus sp. OT, an Acidophilic Sulfate-Reducing Bacterium from Copper Mining Waste in Norilsk, Northern Siberia

    OpenAIRE

    Abicht, Helge K.; Mancini, Stefano; Karnachuk, Olga V.; Solioz, Marc

    2011-01-01

    We have sequenced the genome of Desulfosporosinus sp. OT, a Gram-positive, acidophilic sulfate-reducing Firmicute isolated from copper tailing sediment in the Norilsk mining-smelting area in Northern Siberia, Russia. This represents the first sequenced genome of a Desulfosporosinus species. The genome has a size of 5.7 Mb and encodes 6,222 putative proteins.

  7. Genome sequence of Desulfosporosinus sp. OT, an acidophilic sulfate-reducing bacterium from copper mining waste in Norilsk, Northern Siberia.

    Science.gov (United States)

    Abicht, Helge K; Mancini, Stefano; Karnachuk, Olga V; Solioz, Marc

    2011-11-01

    We have sequenced the genome of Desulfosporosinus sp. OT, a Gram-positive, acidophilic sulfate-reducing Firmicute isolated from copper tailing sediment in the Norilsk mining-smelting area in Northern Siberia, Russia. This represents the first sequenced genome of a Desulfosporosinus species. The genome has a size of 5.7 Mb and encodes 6,222 putative proteins.

  8. Draft Genome Sequence of a Novel Acidophilic Iron-Oxidizing Firmicutes Species, “Acidibacillus ferrooxidans” (SLC66T)

    Science.gov (United States)

    Ñancucheo, Ivan; Oliveira, Renato; Dall’Agnol, Hivana; Johnson, D. Barrie; Grail, Barry; Holanda, Roseanne; Nunes, Gisele Lopes; Cuadros-Orellana, Sara

    2016-01-01

    Here, we present the draft genome sequence of the type strain of “Acidibacillus ferrooxidans,” a mesophilic, heterotrophic, and acidophilic bacterium that was isolated from mine spoilage subjected to accelerated weathering in humidity cell tests carried out by the former U.S. Bureau of Mines in Salt Lake City, UT. PMID:27198020

  9. Draft Genome Sequence of a Novel Acidophilic Iron-Oxidizing Firmicutes Species, "Acidibacillus ferrooxidans" (SLC66T).

    Science.gov (United States)

    Ñancucheo, Ivan; Oliveira, Renato; Dall'Agnol, Hivana; Johnson, D Barrie; Grail, Barry; Holanda, Roseanne; Nunes, Gisele Lopes; Cuadros-Orellana, Sara; Oliveira, Guilherme

    2016-05-19

    Here, we present the draft genome sequence of the type strain of "Acidibacillus ferrooxidans," a mesophilic, heterotrophic, and acidophilic bacterium that was isolated from mine spoilage subjected to accelerated weathering in humidity cell tests carried out by the former U.S. Bureau of Mines in Salt Lake City, UT.

  10. Methanotrophic activity and bacterial diversity in volcanic-geothermal soils at Pantelleria island (Italy)

    Science.gov (United States)

    Gagliano, A. L.; D'Alessandro, W.; Tagliavia, M.; Parello, F.; Quatrini, P.

    2014-04-01

    Verrucomicrobia. Alphaproteobacteria of the genus Methylocystis were isolated from enrichment cultures, under a methane containing atmosphere at 37 °C. The isolates grow at pH 3.5-8 and temperatures of 18-45 °C, and show a methane oxidation rate of ~ 450 μmol mol-1 h-1. Soils from Favara Grande showed the largest diversity of methanotrophic bacteria until now detected in a geothermal soil. While methanotrophic Verrucomicrobia are reported to dominate highly acidic geothermal sites, our results suggest that slightly acidic soils, in high enthalpy geothermal systems, host a more diverse group of both culturable and uncultivated methanotrophs.

  11. Gammaproteobacterial methanotrophs dominate cold methane seeps in floodplains of West Siberian rivers.

    Science.gov (United States)

    Oshkin, Igor Y; Wegner, Carl-Eric; Lüke, Claudia; Glagolev, Mikhail V; Filippov, Illiya V; Pimenov, Nikolay V; Liesack, Werner; Dedysh, Svetlana N

    2014-10-01

    A complex system of muddy fluid-discharging and methane (CH4)-releasing seeps was discovered in a valley of the river Mukhrinskaya, one of the small rivers of the Irtysh Basin, West Siberia. CH4 flux from most (90%) of these gas ebullition sites did not exceed 1.45 g CH4 h(-1), while some seeps emitted up to 5.54 g CH4 h(-1). The δ(13)C value of methane released from these seeps varied between -71.1 and -71.3‰, suggesting its biogenic origin. Although the seeps were characterized by low in situ temperatures (3.5 to 5°C), relatively high rates of methane oxidation (15.5 to 15.9 nmol CH4 ml(-1) day(-1)) were measured in mud samples. Fluorescence in situ hybridization detected 10(7) methanotrophic bacteria (MB) per g of mud (dry weight), which accounted for up to 20.5% of total bacterial cell counts. Most (95.8 to 99.3%) methanotroph cells were type I (gammaproteobacterial) MB. The diversity of methanotrophs in this habitat was further assessed by pyrosequencing of pmoA genes, encoding particulate methane monooxygenase. A total of 53,828 pmoA gene sequences of seep-inhabiting methanotrophs were retrieved and analyzed. Nearly all of these sequences affiliated with type I MB, including the Methylobacter-Methylovulum-Methylosoma group, lake cluster 2, and several as-yet-uncharacterized methanotroph clades. Apparently, microbial communities attenuating methane fluxes from these local but strong CH4 sources in floodplains of high-latitude rivers have a large proportion of potentially novel, psychrotolerant methanotrophs, thereby providing a challenge for future isolation studies. Copyright © 2014, American Society for Microbiology. All Rights Reserved.

  12. Biodegradation of trichloroethylene (TCE) by methanotrophic community.

    Science.gov (United States)

    Shukla, Awadhesh K; Vishwakarma, Pranjali; Upadhyay, S N; Tripathi, Anil K; Prasana, H C; Dubey, Suresh K

    2009-05-01

    Laboratory incubation experiments were carried out to assess the potential of methanotrophic culture for degrading TCE. Measurements of the growth rate and TCE degradation showed that the methanotrophs not only grew in presence of TCE but also degraded TCE. The rate of TCE degradation was found to be 0.19 ppm h(-1). The reverse transcriptase-PCR test was conducted to quantify expression of pmoA and mmoX genes. RT-PCR revealed expression of pmoA gene only. This observation provides evidence that the pmoA gene was functionally active for pMMO enzyme during the study. The diversity of the methanotrophs involved in TCE degradation was assessed by PCR amplification, cloning, restriction fragment length polymorphism and phylogenetic analysis of pmoA genes. Results suggested the occurrence of nine different phylotypes belonging to Type II methanotrophs in the enriched cultures. Out of the nine, five clustered with, genera Methylocystis and rest got clustered in to a separate group.

  13. Modelling the growth of a methanotrophic biofilm

    DEFF Research Database (Denmark)

    Arcangeli, J.-P.; Arvin, E.

    1999-01-01

    that heterotrophs and nitrifiers co-existed with methanotrophs in the biofilm. Heterotrophic biomass grew on soluble polymers formed by the hydrolysis of dead biomass entrapped in the biofilm. Nitrifier populations developed because of the presence of ammonia in the mineral medium. Based on these experimental...

  14. Identification of Methanotrophic Biomarker Lipids in the Symbiont-Containing Gills of Seep Mussels

    Science.gov (United States)

    Jahnke, L. L.; Zahiralis, K. D.; Klein, H. P.; Morrison, David (Technical Monitor)

    1994-01-01

    Mussels collected from hydrocarbon seeps in the Gulf of Mexico grow with methane as sole carbon and energy source due to a symbiotic association with methane-oxidizing bacteria. Transmission electron micrographs of mussel gills show cells with stacked intracytoplasmic membranes similar to type I methanotrophic bacteria. Methanotrophs are known to synthesize several types of cyclic triterpenes, hopanoids and methyl sterols, as well as unique monounsaturated fatty acid, double bond positional isomers that serve as biomarkers for this group. Lipid analysis of dissected mussels demonstrated the presence of these biomarkers predominantly in the gill tissue with much smaller amounts in mantle and remaining body tissues. Gill tissue contained 1150 micrograms/g dry wt. of hopanepolyol derivatives and diplopterol while the mantle tissue contained only 17 micrograms/g. The C16 monounsaturated fatty acids (16:1) characteristic of type I methanotrophic membranes dominated the gill tissue making up 53% of the total while only 5% 16:1 was present in the mantle tissue. The methyl sterol distribution was more dispersed. The predominant sterol in all tissues was cholesterol with lesser amounts of other desmethyl and 4-methyl sterols. The gill and mantle tissues contained 3461 micrograms (17% methyl) and 2750 micrograms (5% methyl) sterol per gm dry wt., respectively. Methyl sterol accounted for 44% of the sterol esters isolated from the gill, suggesting active demethylation of the methanotrophic sterols in this tissue. The use of lipid biomarkers could provide an effective means for identifying host-symbiont relationships.

  15. Mineral Salt Medium (MSM) for extreme acidophilic microorganisms

    OpenAIRE

    sprotocols

    2015-01-01

    Medium for growth of extreme acidophilic microorganisms. This medium does not contain trace elements. When not working on mineral, addition of trace element (TE) solution is necessary, see separate protocol. http://www.nature.com/protocolexchange/protocols/3811

  16. High diversity of methanotrophic bacteria in geothermal soils affected by high methane fluxes

    Science.gov (United States)

    D'Alessandro, Walter; Gagliano, Antonina Lisa; Quatrini, Paola; Parello, Francesco

    2014-05-01

    Volcanic and geothermal systems emit endogenous gases by widespread degassing from soils, including CH4, a greenhouse gas 25 times as potent as CO2. Recently, it has been demonstrated that volcanic/geothermal soils act as source, but also as biological filter for methane release to the atmosphere. For long time, volcanic/geothermal soils has been considered inhospitable for methanotrophic microorganisms, but new extremophile methanotrophs belonging to Verrucomicrobia were identified in three different areas (Pozzuoli, Italy; Hell's Gate, New Zealand; Kamchatka, Russia), explaining anomalous behaviours in methane leakages of several geothermal/volcanic sites. Our aim was to increase the knowledge of the relationship between methane emissions from volcanic/geothermal areas and biological methane oxidation, by investigating a geothermal site of Pantelleria island (Italy). Pantelleria Island hosts a high enthalpy geothermal system characterized by high temperature, high CH4 and very low H2S fluxes. Such characteristics are reflected in potentially great supply of methane for methanotrophs and scarce presence of inhibitors of their activity (H2S and NH3) in the Pantelleria soils. Potential methanotrophic activity within these soils was already evidenced by the CH4/CO2 ratio of the flux measurements which was lower than that of the respective fumarolic manifestations indicating a loss of CH4 during the gas travel towards the earth's surface. In this study laboratory incubation experiments using soils sampled at Favara Grande, the main hydrothermal area of Pantelleria, showed very high methane consumption rates (up to 9500 ng CH4 h-1 g-1). Furthermore, microbiological and culture-independent molecular analyses allowed to detect the presence of methanotrophs affiliated to Gamma- and Alpha-Proteobacteria and to the newly discovered acidothermophilic methanotrophs Verrucomicrobia. Culturable methanotrophic Alpha-proteobacteria of the genus Methylocystis were isolated by

  17. Diversity of methanotroph communities in a basalt aquifer.

    Science.gov (United States)

    Newby, D T; Reed, D W; Petzke, L M; Igoe, A L; Delwiche, M E; Roberto, F F; McKinley, J P; Whiticar, M J; Colwell, F S

    2004-06-01

    Methanotrophic bacteria play an important role in global cycling of carbon and co-metabolism of contaminants. Methanotrophs from pristine regions of the Snake River Plain Aquifer (SRPA; Idaho, USA) were studied in order to gain insight into the native groundwater communities' genetic potential to carry out TCE co-metabolism. Wells were selected that were proximal to a TCE plume believed to be undergoing natural attenuation. Methane concentrations ranged from 1 to >1000 nM. Carbon isotope ratios and diversity data together suggest that the SRPA contains active communities of methanotrophs that oxidize microbially produced methane. Microorganisms removed from groundwater by filtration were used as inocula for enrichments or frozen immediately and DNA was subsequently extracted for molecular characterization. Primers that specifically target methanotroph 16S rRNA genes or genes that code for subunits of soluble or particulate methane monooxygenase, mmoX and pmoA, respectively, were used to characterize the indigenous methanotrophs via PCR, cloning, RFLP analysis, and sequencing. Type I methanotroph clones aligned with Methylomonas, Methylocaldum, and Methylobacter sequences and a distinct 16S rRNA phylogenetic lineage grouped near Methylobacter. The majority of clone sequences in type II methanotroph 16S rRNA, pmoA, and mmoX gene libraries grouped closely with sequences in the Methylocystis genus. A subset of the type II methanotroph clones from the aquifer had sequences that aligned most closely to Methylosinus trichosporium OB3b and Methylocystis spp., known TCE-co-metabolizing methanotrophs.

  18. Diversity of methanotrophs in a simulated modified biocover reactor

    Institute of Scientific and Technical Information of China (English)

    Zifang Chi; Wenjing Lu; Hongtao Wang; Yan Zhao

    2012-01-01

    A simulated landfill biocover microcosm consisting of a modifying ceramsite material and compost were investigated.Results show that the mixture can improve the material porosity and achieve a stable and highly efficient (100%) methane oxidation over an extended operating period.The diversity of the methanotrophic community in the microcosm was assessed.Type Ⅰ methanotrophs were enhanced in the microcosm due to the increased air diffusion and distribution,whereas the microbial diversity and population density of type Ⅱ methanotrophs were not significantly affected.Moreover,the type Ⅰ methanotrophic community structure significantly varied with the reactor height,whereas that of type Ⅱ methanotrophic communities did not exhibit a spatial variation.Phylogenetic analysis showed that type Ⅰ methanotroph-based nested polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE) resulted in the detection of eight different populations,most of which are related to Methylobacter sp.,whereas that of type Ⅱ resulted in the detection of nine different populations,most of which are related to Methylocystaceae.Methanotrophic community analysis also indicated that a number of new methanotrophic genera not closely related to any known methanotrophic populations were present.

  19. Methane and trichloroethylene oxidation by an estuarine methanotroph, Methylobacter sp. strain BB5.1.

    OpenAIRE

    Smith, K. S.; Costello, A. M.; Lidstrom, M E

    1997-01-01

    An estuarine methanotroph was isolated from sediment enrichments and designated Methylobacter sp. strain BB5.1. In cells grown on medium with added copper, oxidation of methane and trichloroethylene occurred with similar Ks values, but the Vmax for trichloroethylene oxidation was only 0.1% of the methane oxidation Vmax. Cells grown on low-copper medium did not oxidize trichloroethylene and showed a variable rate of methane oxidation.

  20. Recovery of paddy soil methanotrophs from long term drought

    NARCIS (Netherlands)

    Collet, S.; Reim, A.; Ho, A.; Frenzel, P.

    2015-01-01

    Air-dried paddy soils stored for 1–18 years were used to examine the resistance of methanotrophs to drought. Older air-dried soils representing longer-lasting drought events reduced methanotrophic diversity, and adversely affected methane oxidation rate after re-wetting. In early incubations the typ

  1. Exploration and prediction of interactions between methanotrophs and heterotrophs.

    Science.gov (United States)

    Stock, Michiel; Hoefman, Sven; Kerckhof, Frederiek-Maarten; Boon, Nico; De Vos, Paul; De Baets, Bernard; Heylen, Kim; Waegeman, Willem

    2013-12-01

    Methanotrophs can form the basis of a methane-driven food web on which heterotrophic microorganisms can feed. In return, these heterotrophs can stimulate growth of methanotrophs in co-culture by providing growth additives. However, only a few specific interactions are currently known. We incubated nine methanotrophs with 25 heterotrophic strains in a pairwise miniaturized co-cultivation setup. Through principal component analysis and k-means clustering, methanotrophs and heterotrophs could be grouped according to their interaction behaviour, suggesting strain-dependent methanotroph-heterotroph complementarity. Co-cultivation significantly enhanced the growth parameters of three methanotrophs. This was most pronounced for Methylomonas sp. M5, with a threefold increase in maximum density and a fourfold increase in maximum increase in density in co-culture with Cupriavidus taiwanensis LMG 19424. In contrast, co-cultivation with Methylobacterium radiotolerans LMG 2269 and Pseudomonas aeruginosa LMG 12228 inhibited growth of most methanotrophs. Functional genomic analysis suggested the importance of vitamin metabolism for co-cultivation success. The generated data set was then successfully exploited as a proof-of-principle for predictive modelling of co-culture responses based on other interactions of the same heterotrophs and methanotrophs, yielding values of the area under the receiver operating characteristic curve of 0.73 upon 50% missing values for the maximum increase in density parameter. As such, these modelling-based tools were shown to hold great promise in reducing the amount of data that needs to be generated when conducting large co-cultivation studies.

  2. Final Technical Report: Role of Methanotrophs in Metal Mobilization, Metal Immobilization and Mineral Weathering: Effects on the In Situ Microbial Community and the Sustainability of Subsurface Water Systems

    Energy Technology Data Exchange (ETDEWEB)

    Semrau, Jeremy D. [Univ. of Michigan, Ann Arbor, MI (United States); DiSpirito, Alan A. [Iowa State Univ., Ames, IA (United States)

    2016-11-06

    Activities from this DOE sponsored project can be divided into four broad areas: (1) investigations into the potential of methanobactin, a biogenic metal-binding agent produced by methanotrophs, in mitigating mercury toxicity; (2) elucidation of the genetic basis for methanobactin synthesis from methanotrophs; (3) examination of differential gene expression of M. trichosporium OB3b when grown in the presence of varying amounts of copper and/or cerium, and (4) collection and characterization of soil cores from Savannah River Test Site to determine the ubiquity of methanobactin producing methanotrophs. From these efforts, we have conclusively shown that methanobactin can strongly bind mercury as Hg[II], and in so doing significantly reduce the toxicity of this metal to microbes. Further, we have deduced the genetic basis of methanobactin production in methanotrophs, enabling us to construct mutants such that we can now ascribe function to different genes as well as propose a pathway for methanobactin biosynthesis. We have also clear evidence that copper and cerium (as an example of a rare earth element) dramatically affect gene expression in methanotrophs, and thus have an important impact on the activity and application of these microbes to a variety of environmental and industrial issues. Finally, we successfully isolated one methanotroph from the deep subsurface of the Savannah River Test Site and characterized the ability of different forms of methanobactin to mobilize copper and mercury from these soils.

  3. Flow cytometry based techniques to study testicular acidophilic granulocytes from the protandrous fish gilthead seabream (Sparus aurata L.

    Directory of Open Access Journals (Sweden)

    Chaves-Pozo Elena

    2004-01-01

    Full Text Available The gilthead seabream is a protandrous seasonal breeding teleost that is an excellent model for studying the testicular regression process which occurs in both seasonal testicular involution and sex reversion. Little is known about the cell types and the molecular mechanisms involved in such processes, mainly because of the lack of appropriate methods for testis dissociation, and testicular cell isolation, culture and functional characterization. We have previously reported that gilthead seabream acidophilic granulocytes infiltrate the testis at post-spawning stage, settle close to the spermatogonia and accumulate intracellular interleukin-1&bgr;. In this paper, we report several flow cytometry based assays which allow to establish the role played by gilthead seabream testicular acidophilic granulocytes and permits their quantification.

  4. Ardrea characterisation of acidophilic micro-organisms isolated from gold mines in Marmato, Colombia Caracterización por Ardrea de microorganismos acidófilos aislados en minas de oro de Marmato, Colombia

    OpenAIRE

    Osorio E. Victor Manuel; Márquez G. Marco; Márquez F. Edna Judith

    2007-01-01

    Mineral bio-oxidation improves the extraction of valuable metals and also decreases the impact caused by mining waste; however, the interactions between the micro-organisms so involved are little known. Double-layer solid culture media techniques and amplified ribosomal DNA restriction enzyme analysis (Ardrea), using Eco72I, Eco24I, XcmI and BsaAI enzymes, were used for characterising four micro-organisms isolated from gold mines located in Marmato, Colombia. This work was aimed at better und...

  5. Life at acidic pH imposes an increased energetic cost for a eukaryotic acidophile.

    Science.gov (United States)

    Messerli, Mark A; Amaral-Zettler, Linda A; Zettler, Erik; Jung, Sung-Kwon; Smith, Peter J S; Sogin, Mitchell L

    2005-07-01

    Organisms growing in acidic environments, pHpH. We begin to investigate this premise by determining the magnitude of the transmembrane electrochemical H+ gradient in an acidophilic Chlamydomonas sp. (ATCC PRA-125) isolated from the Rio Tinto, a heavy metal laden, acidic river (pH 1.7-2.5). This acidophile grows most rapidly at pH 2 but is capable of growth over a wide pH range (1.5-7.0), while Chlamydomonas reinhardtii is restricted to growth at pH>or=3 with optimal growth between pH 5.5 and 8.5. With the fluorescent H+ indicator, 2',7'-bis-(2-carboxyethyl)-5-(and-6)-carboxyfluorescein (BCECF), we show that the acidophilic Chlamydomonas maintains an average cytosolic pH of 6.6 in culture medium at both pH 2 and pH 7 while Chlamydomonas reinhardtii maintains an average cytosolic pH of 7.1 in pH 7 culture medium. The transmembrane electric potential difference of Chlamydomonas sp., measured using intracellular electrodes at both pH 2 and 7, is close to 0 mV, a rare value for plants, animals and protists. The 40,000-fold difference in [H+] could be the result of either active or passive mechanisms. Evidence for active maintenance was detected by monitoring the rate of ATP consumption. At the peak, cells consume about 7% more ATP per second in medium at pH 2 than at pH 7. This increased rate of consumption is sufficient to account for removal of H+ entering the cytosol across a membrane with relatively high permeability to H+ (7x10(-8) cm s-1). Our results indicate that the small increase in the rate of ATP consumption can account for maintenance of the transmembrane H+ gradient without the imposition of cell surface H+ barriers.

  6. A novel approach for rapidly and cost-effectively assessing toxicity of toxic metals in acidic water using an acidophilic iron-oxidizing biosensor.

    Science.gov (United States)

    Yang, Shih-Hung; Cheng, Kuo-Chih; Liao, Vivian Hsiu-Chuan

    2017-11-01

    Contamination by heavy metals and metalloids is a serious environmental and health concern. Acidic wastewaters are often associated with toxic metals which may enter and spread into agricultural soils. Several biological assays have been developed to detect toxic metals; however, most of them can only detect toxic metals in a neutral pH, not in an acidic environment. In this study, an acidophilic iron-oxidizing bacterium (IOB) Strain Y10 was isolated, characterized, and used to detect toxic metals toxicity in acidic water at pH 2.5. The colorimetric acidophilic IOB biosensor was based on the inhibition of the iron oxidizing ability of Strain Y10, an acidophilic iron-oxidizing bacterium, by metals toxicity. Our results showed that Strain Y10 is acidophilic iron-oxidizing bacterium. Thiobacillus caldus medium (TCM) (pH 2.5) supplied with both S4O6(2-) and glucose was the optimum growth medium for Strain Y10. The optimum temperature and pH for the growth of Strain Y10 was 45 °C and pH 2.5, respectively. Our study demonstrates that the color-based acidophilic IOB biosensor can be semi-quantitatively observed by eye or quantitatively measured by spectrometer to detect toxicity from multiple toxic metals at pH 2.5 within 45 min. Our study shows that monitoring toxic metals in acidic water is possible by using the acidophilic IOB biosensor. Our study thus provides a novel approach for rapid and cost-effective detection of toxic metals in acidic conditions that can otherwise compromise current methods of chemical analysis. This method also allows for increased efficiency when screening large numbers of environmental samples. Copyright © 2017 Elsevier Ltd. All rights reserved.

  7. [Methanotrophic bacteria in cold seeps of the floodplains of northern rivers].

    Science.gov (United States)

    Belova, S É; Oshkin, I Iu; Glagolev, M V; Lapshina, E D; Maksiutov, Sh Sh; Dedysh, S N

    2013-01-01

    Small mud volcanoes (cold seeps), which are common in the floodplains of northern rivers, are a potentially important, although poorly studied sources of atmospheric methane. Field research on the cold seeps of the Mukhrina River (Khanty-Mansiysk Autonomous okrug, Russia) revealed methane fluxes from these structures to be orders of magnitude higher than from equivalent areas of the mid-taiga bogs. Microbial communities developing around the seeps were formed under conditions of high methane concentrations, low temperatures (3-5 degrees C), and near-neutral pH. Molecular identification of methane-oxidizing bacteria from this community by analysis of the pmoA gene encoding particulate methane monooxygenase revealed both type I and type II methanotrophs (classes Gammaproteobacteria and Alphaproteobacteria, respectively), with predomination of type I methanotrophs. Among the latter, microorganisms related to Methylobacterpsychrophilus and Methylobacter tundripaludum, Crenothrix polyspora (a stagnant water dweller), and a number of methanotrophs belonging to unknown taxa were detected. Growth characteristics of two isolates were determined. Methylobactersp. CMS7 exhibited active growth at 4-10 degrees C, while Methylocystis sp. SB12 grew better at 20 degrees C. Experimental results confirmed the major role ofmethanotrophic gammaproteobacteria in controlling the methane emission from cold river seeps.

  8. Genomics of the thermo-acidophilic red alga Galdieria sulphuraria

    OpenAIRE

    Barbier, Guillaume G.; Weber, Andreas P. M.; Zimmer, Marc

    2005-01-01

    Extremophilic organisms dwell in environments that are characterized by high or low temperatures (thermophiles or psychrophiles), very low or high pH-values (acidophiles or alkalophiles), high salt concentrations (halophiles), high pressure (barophiles), or extreme drought (xerophiles). Many extremophiles are microbes, and many also belong to the prokaryota. Galdieria sulphuraria, however, is a member of a group of extremophilic eukaryotes that are named Cyanidiales. Cyanidiales are unicellul...

  9. Draft Genome Sequence of Methylocaldum sp. SAD2, a Methanotrophic Strain That Can Convert Raw Biogas to Methanol in the Presence of Hydrogen Sulfide.

    Science.gov (United States)

    Wei, Xiangdong; Ge, Xumen; Li, Yebo; Yu, Zhongtang

    2017-08-10

    The draft genome sequence of Methylocaldum sp. SAD2, a methanotrophic strain isolated from a hydrogen sulfide-rich anaerobic digester, is reported here. Strain SAD2 possesses genes for methane oxidation in the presence of H2S. Copyright © 2017 Wei et al.

  10. Draft Genome Sequence of Methylocaldum sp. Strain 14B, an Obligate Hydrogen Sulfide-Tolerant Methanotrophic Strain That Can Convert Biogas to Methanol

    Science.gov (United States)

    Wei, Xiangdong; Ge, Xumen; Li, Yebo

    2017-01-01

    ABSTRACT The draft genome sequence of Methylocaldum sp. 14B, an obligate methanotrophic strain isolated from solid-state anaerobic digestion systems, is reported here. Strain 14B possesses genes for methane oxidation and exhibited tolerance to H2S. PMID:28428289

  11. Reverse Methanogenesis and Respiration in Methanotrophic Archaea

    Directory of Open Access Journals (Sweden)

    Peer H. A. Timmers

    2017-01-01

    Full Text Available Anaerobic oxidation of methane (AOM is catalyzed by anaerobic methane-oxidizing archaea (ANME via a reverse and modified methanogenesis pathway. Methanogens can also reverse the methanogenesis pathway to oxidize methane, but only during net methane production (i.e., “trace methane oxidation”. In turn, ANME can produce methane, but only during net methane oxidation (i.e., enzymatic back flux. Net AOM is exergonic when coupled to an external electron acceptor such as sulfate (ANME-1, ANME-2abc, and ANME-3, nitrate (ANME-2d, or metal (oxides. In this review, the reversibility of the methanogenesis pathway and essential differences between ANME and methanogens are described by combining published information with domain based (metagenome comparison of archaeal methanotrophs and selected archaea. These differences include abundances and special structure of methyl coenzyme M reductase and of multiheme cytochromes and the presence of menaquinones or methanophenazines. ANME-2a and ANME-2d can use electron acceptors other than sulfate or nitrate for AOM, respectively. Environmental studies suggest that ANME-2d are also involved in sulfate-dependent AOM. ANME-1 seem to use a different mechanism for disposal of electrons and possibly are less versatile in electron acceptors use than ANME-2. Future research will shed light on the molecular basis of reversal of the methanogenic pathway and electron transfer in different ANME types.

  12. Reverse Methanogenesis and Respiration in Methanotrophic Archaea

    Science.gov (United States)

    Koehorst, Jasper J.; Jetten, Mike S. M.; Stams, Alfons J. M.

    2017-01-01

    Anaerobic oxidation of methane (AOM) is catalyzed by anaerobic methane-oxidizing archaea (ANME) via a reverse and modified methanogenesis pathway. Methanogens can also reverse the methanogenesis pathway to oxidize methane, but only during net methane production (i.e., “trace methane oxidation”). In turn, ANME can produce methane, but only during net methane oxidation (i.e., enzymatic back flux). Net AOM is exergonic when coupled to an external electron acceptor such as sulfate (ANME-1, ANME-2abc, and ANME-3), nitrate (ANME-2d), or metal (oxides). In this review, the reversibility of the methanogenesis pathway and essential differences between ANME and methanogens are described by combining published information with domain based (meta)genome comparison of archaeal methanotrophs and selected archaea. These differences include abundances and special structure of methyl coenzyme M reductase and of multiheme cytochromes and the presence of menaquinones or methanophenazines. ANME-2a and ANME-2d can use electron acceptors other than sulfate or nitrate for AOM, respectively. Environmental studies suggest that ANME-2d are also involved in sulfate-dependent AOM. ANME-1 seem to use a different mechanism for disposal of electrons and possibly are less versatile in electron acceptors use than ANME-2. Future research will shed light on the molecular basis of reversal of the methanogenic pathway and electron transfer in different ANME types. PMID:28154498

  13. Environmental evidence for net methane production and oxidation in putative ANaerobic MEthanotrophic (ANME) archaea

    DEFF Research Database (Denmark)

    Lloyd, Karen; Teske, Andreas; Alperin, Marc J.

    2011-01-01

    Uncultured ANaerobic MEthanotrophic (ANME) archaea are often assumed to be obligate methanotrophs that are incapable of net methanogenesis, and are therefore used as proxies for anaerobic methane oxidation in many environments in spite of uncertainty regarding their metabolic capabilities...

  14. Gene loss and horizontal gene transfer contributed to the genome evolution of the extreme acidophile Ferrovum

    Directory of Open Access Journals (Sweden)

    Sophie Roxana Ullrich

    2016-05-01

    Full Text Available Acid mine drainage (AMD, associated with active and abandoned mining sites, is a habitat for acidophilic microorganisms that gain energy from the oxidation of reduced sulfur compounds and ferrous iron and that thrive at pH below 4. Members of the recently proposed genus Ferrovum are the first acidophilic iron oxidizers to be described within the Betaproteobacteria. Although they have been detected as typical community members in AMD habitats worldwide, knowledge of their phylogenetic and metabolic diversity is scarce. Genomics approaches appear to be most promising in addressing this lacuna since isolation and cultivation of Ferrovum has proven to be extremely difficult and has so far only been successful for the designated type strain Ferrovum myxofaciens P3G. In this study, the genomes of two novel strains of Ferrovum (PN-J185 and Z-31 derived from water samples of a mine water treatment plant were sequenced. These genomes were compared with those of Ferrovum sp. JA12 that also originated from the mine water treatment plant, and of the type strain (P3G. Phylogenomic scrutiny suggests that the four strains represent three Ferrovum species that cluster in two groups (1 and 2. Comprehensive analysis of their predicted metabolic pathways revealed that these groups harbor characteristic metabolic profiles, notably with respect to motility, chemotaxis, nitrogen metabolism, biofilm formation and their potential strategies to cope with the acidic environment. For example, while the F. myxofaciens strains (group 1 appear to be motile and diazotrophic, the non-motile group 2 strains have the predicted potential to use a greater variety of fixed nitrogen sources. Furthermore, analysis of their genome synteny provides first insights into their genome evolution, suggesting that horizontal gene transfer and genome reduction in the group 2 strains by loss of genes encoding complete metabolic pathways or physiological features contributed to the observed

  15. Streptacidiphilus gen. nov., acidophilic actinomycetes with wall chemotype I and emendation of the family Streptomycetaceae (Waksman and Henrici (1943)AL) emend. Rainey et al. 1997.

    Science.gov (United States)

    Kim, Seung Bum; Lonsdale, John; Seong, Chi-Nam; Goodfellow, Michael

    2003-01-01

    The taxonomic position of acidophilic actinomycetes selectively isolated from acidic soils and litter was examined using a polyphasic approach. The distinct 16S rDNA phyletic branch formed by representative strains was equated with related monophyletic clades that corresponded to the genera Kitasatospora and Streptomyces. The acidophilic isolates also exhibited a distinctive pH profile, a unique 16S rDNA signature, and contained major amounts of LL-diaminopimelic acid, galactose and rhamnose in whole-organism hydrolysates. It is proposed that these acidophilic actinomycetes be assigned to a new genus, Streptacidiphilus gen. nov., on the basis of genotypic and phenotypic differences. Three species were defined on the basis of DNA:DNA pairing and phenotypic data, namely, Streptacidiphilus albus sp. nov., the type species, Streptacidiphilus neutrinimicus sp. nov. and Streptacidiphilus carbonis sp. nov. Members of the genera Kitasatospora, Streptacidiphilus and Streptomyces share a number of key characteristics and form a stable monophyletic branch in the 16S rDNA tree. It is, therefore, proposed that the description of the family Streptomycetaceae be emended to account for properties shown by Kitasatospora and Streptacidiphilus species.

  16. Aquatic plant surface as a niche for methanotrophs

    Directory of Open Access Journals (Sweden)

    Naoko eYoshida

    2014-02-01

    Full Text Available This study investigated the potential local CH4 sink in various plant parts as a boundary environment of CH4 emission and consumption. By comparing CH4 consumption activities in cultures inoculated with parts from 39 plant species, we observed significantly higher consumption of CH4 associated with aquatic plants than other emergent plant parts such as woody plant leaves, macrophytic marine algae, and sea grass. In situ activity of CH4 consumption by methanotrophs associated with different species of aquatic plants was in the range of 3.7 – 37 μmol⋅h-1⋅g-1 dry weight, which was ca 5.7-370 fold higher than epiphytic CH4 consumption in submerged parts of emergent plants. The qPCR-estimated copy numbers of the particulate methane monooxygenase-encoding gene pmoA were variable among the aquatic plants and ranged in the order of 105 to 107 copies⋅g-1 dry weight, which correlated with the observed CH4 consumption activities. Phylogenetic identification of methanotrophs on aquatic plants based on the pmoA sequence analysis revealed a predominance of diverse gammaproteobacterial type-I methanotrophs, including a phylotype of a possible plant-associated methanotroph with the closest identity (86-89% to Methylocaldum gracile.

  17. Enrichment of mesophilic acidophiles from the Underground Copper Mine Bor

    Directory of Open Access Journals (Sweden)

    Conić Vesna T.

    2009-01-01

    Full Text Available In this work, autotrophic growth of mesophilic acidophiles from the Underground Copper Mine Bor was performed. Two selected solution samples collected from the 'Tilva Roš' ore body were prepared in a 9K nutrient medium (Silverman and Lundgren, 1959. The first sample TR k-16 was obtained during the hole drilling of the ore body, and the second TR k-31 from the drainage channel. Two samples of 9K media (Silverman and Lundgren, 1959 were inoculated with two selected solution samples from the underground mine Tilva Roš. Inoculated culture media were incubated without prior autoclaving in the period of 6 days at a temperature of 28 ?C with purging air through the system with enough oxygen and carbon dioxide. Oxidation rate of ferrous ions in the first 3 days of incubation was 14.8 and 10.7 wt.% Fe2+/day, the next 3 days 17.3 and 13.6 and for the total period of 6 days 98.3 and 74.8 wt.% for the first and second sample, respectively, i.e. 100 wt.% with initial percentage of ferrous ion in each medium. After centrifugation of enriched samples of culture media at 3000 rpm for 5 min, a plenty of mesophilic acidophiles were determined by microscopic method. According to Karavaiko [6], in the processes of incubition for 9K nutrient solution cells number reach a value of 108 cells/cm3.

  18. Methane oxidation by an extremely acidophilic bacterium of the phylum Verrucomicrobia

    NARCIS (Netherlands)

    Dunfield, P.F.; Yurgey, A.Q.; Senin, P.; Smirnova, A.V.; Stott, M.B.; Hou, S.; Ly, B.; Saw, J.H.; Zhou, Z.; Ren, Y.; Wang, J.; Mountain, B.W.; Crowe, M.A.; Weatherby, T.M.; Bodelier, P.L.E.; Liesack, W.; Feng, L.; Wang, L.; Alam, M.

    2007-01-01

    Aerobic methanotrophic bacteria consume methane as it diffuses away from methanogenic zones of soil and sediment1. They act as a biofilter to reduce methane emissions to the atmosphere, and they are therefore targets in strategies to combat global climate change. No cultured methanotroph grows optim

  19. Effect of afforestation and reforestation of pastures on the activity and population dynamics of methanotrophic bacteria.

    Science.gov (United States)

    Singh, Brajesh K; Tate, Kevin R; Kolipaka, Gokul; Hedley, Carolyn B; Macdonald, Catriona A; Millard, Peter; Murrell, J Colin

    2007-08-01

    We investigated the effect of afforestation and reforestation of pastures on methane oxidation and the methanotrophic communities in soils from three different New Zealand sites. Methane oxidation was measured in soils from two pine (Pinus radiata) forests and one shrubland (mainly Kunzea ericoides var. ericoides) and three adjacent permanent pastures. The methane oxidation rate was consistently higher in the pine forest or shrubland soils than in the adjacent pasture soils. A combination of phospholipid fatty acid (PLFA) and stable isotope probing (SIP) analyses of these soils revealed that different methanotrophic communities were active in soils under the different vegetations. The C18 PLFAs (signature of type II methanotrophs) predominated under pine and shrublands, and C16 PLFAs (type I methanotrophs) predominated under pastures. Analysis of the methanotrophs by molecular methods revealed further differences in methanotrophic community structure under the different vegetation types. Cloning and sequencing and terminal-restriction fragment length polymorphism analysis of the particulate methane oxygenase gene (pmoA) from different samples confirmed the PLFA-SIP results that methanotrophic bacteria related to type II methanotrophs were dominant in pine forest and shrubland, and type I methanotrophs (related to Methylococcus capsulatus) were dominant in all pasture soils. We report that afforestation and reforestation of pastures caused changes in methane oxidation by altering the community structure of methanotrophic bacteria in these soils.

  20. The taxonomic and physiologic diversity of the acidophilic bacteria of the genus Thiobacillus used in ores solubilization processes

    Directory of Open Access Journals (Sweden)

    Carmen Mădălina Cişmaşiu

    2010-01-01

    Full Text Available The development of biotechnological processes, based mainly on the activity of the acidophilic chemolithotrophic, proved their efficiency in recovering metals from sulphides ores and mining drains and in bioremediation of the polluted environment with residual inorganic substances, like the heavy metals ions and their compounds.Due to the influence of the physical-chemical factors on the development and the metabolic activity of the microorganism’s present in the industrial effluents, the study of these parameters was imposed for raising the efficiency of the processes of adsorption and biosolubilization of the metallic ions. A special importance for using bacteria of the genus Acidithiobacillus in the biosolubilization processes of heavy metals from acid mine tailings is represented by the resistance of these bacteria to high concentrations of metal ions.The experiments prove a strong relationship between the acidity of the medium and the behaviour of the acidophilic chemolithotrophic bacteria. The comparative analyses regarding the influence of metallic ions (Cu2+, Zn2+ and Fe2+ on the physiologic diversity of the Acidithiobacillus populations, isolated from the mining sites, demonstrated the higher resistance of these bacteria to higher concentrations of metallic ions.

  1. Iron meteorites can support the growth of acidophilic chemolithoautotrophic microorganisms.

    Science.gov (United States)

    González-Toril, Elena; Martínez-Frías, Jesús; Gómez Gómez, José María; Rull, Fernando; Amils, Ricardo

    2005-06-01

    Chemolithoautotrophy based on reduced inorganic minerals is considered a primitive energy transduction system. Evidence that a high number of meteorites crashed into the planet during the early period of Earth history led us to test the ability of iron-oxidizing bacteria to grow using iron meteorites as their source of energy. Here we report the growth of two acidophilic iron-oxidizing bacteria, Leptospirillum ferrooxidans and Acidithiobacillus ferrooxidans, on a piece of the Toluca meteorite as the only source of energy. The alteration of the surface of the exposed piece of meteorite, the solubilization of its oxidized metal constituents, mainly ferric iron, and the formation of goethite precipitates all clearly indicate that iron-meteorite-based chemolithotrophic metabolism is viable.

  2. Modelling of the growth of a methanotrophic biofilm

    DEFF Research Database (Denmark)

    Arcangeli, J.-P.; Arvin, E.

    1997-01-01

    . It indicated that the most influential factors were those related to the biofilm (i.e. density; solid volume fraction; thickness). This suggests that in order to improve the model, further research is needed in the field of biofilm structure and composition. (C) 1997 IAWQ. Published by Elsevier Science Ltd.......A model describing the growth of a methanotrophic biofilm is presented. This model involves simultaneous growth of methanotrophs, heterotrophs and nitrifiers. Heterotrophic biomass grows on soluble polymers which arise from the hydrolysis of dead biomass entrapped in the biofilm. Nitrifiers develop...... because of the presence of ammonia in the mineral medium. A comparison of this model with experimental data showed that the biofilm growth, methane removal, oxygen consumption, product formation and biofilm detachment could be fitted well. Parameter estimation yielded a maximum growth rate...

  3. Electricity generation from an inorganic sulfur compound containing mining wastewater by acidophilic microorganisms

    NARCIS (Netherlands)

    Ni, Gaofeng; Christel, Stephan; Roman, Pawel; Wong, Zhen Lim; Bijmans, Martijn F.M.; Dopson, Mark

    2016-01-01

    Sulfide mineral processing often produces large quantities of wastewaters containing acid-generating inorganic sulfur compounds. If released untreated, these wastewaters can cause catastrophic environmental damage. In this study, microbial fuel cells were inoculated with acidophilic

  4. Electricity generation from an inorganic sulfur compound containing mining wastewater by acidophilic microorganisms

    NARCIS (Netherlands)

    Ni, Gaofeng; Christel, Stephan; Roman, Pawel; Wong, Zhen Lim; Bijmans, Martijn F.M.; Dopson, Mark

    2016-01-01

    Sulfide mineral processing often produces large quantities of wastewaters containing acid-generating inorganic sulfur compounds. If released untreated, these wastewaters can cause catastrophic environmental damage. In this study, microbial fuel cells were inoculated with acidophilic microorganism

  5. Recurrence and frequency of disturbance have cumulative effect on methanotrophic activity, abundance, and community structure.

    Directory of Open Access Journals (Sweden)

    Adrian eHo

    2016-01-01

    Full Text Available Alternate prolonged drought and heavy rainfall is predicted to intensify with global warming. Desiccation-rewetting events alter the soil quality and nutrient concentrations which drive microbial-mediated processes, including methane oxidation, a key biogeochemical process catalyzed by methanotrophic bacteria. Although aerobic methanotrophs showed remarkable resilience to a suite of physical disturbances induced as a single event, their resilience to recurring disturbances is less known. Here, using a rice field soil in a microcosm study, we determined whether recurrence and frequency of desiccation-rewetting impose an accumulating effect on the methanotrophic activity. The response of key aerobic methanotroph subgroups (type Ia, Ib, and II were monitored using qPCR assays, and was supported by a t-RFLP analysis. The methanotrophic activity was resilient to recurring desiccation-rewetting, but increasing the frequency of the disturbance by two-fold significantly decreased methane uptake rate. Both the qPCR and t-RFLP analyses were congruent, showing the dominance of type Ia/Ib methanotrophs prior to disturbance, and after disturbance, the recovering community was predominantly comprised of type Ia (Methylobacter methanotrophs. Both type Ib and type II (Methylosinus/Methylocystis methanotrophs were adversely affected by the disturbance, but type II methanotrophs showed recovery over time, indicating relatively higher resilience to the disturbance. This revealed distinct, yet unrecognized traits among the methanotroph community members. Our results show that recurring desiccation-rewetting before a recovery in community abundance had an accumulated effect, compromising methanotrophic activity. While methanotrophs may recover well following sporadic disturbances, their resilience may reach a ‘tipping point’ where activity no longer recovered if disturbance persists and increase in frequency.

  6. Recurrence and Frequency of Disturbance have Cumulative Effect on Methanotrophic Activity, Abundance, and Community Structure.

    Science.gov (United States)

    Ho, Adrian; van den Brink, Erik; Reim, Andreas; Krause, Sascha M B; Bodelier, Paul L E

    2015-01-01

    Alternate prolonged drought and heavy rainfall is predicted to intensify with global warming. Desiccation-rewetting events alter the soil quality and nutrient concentrations which drive microbial-mediated processes, including methane oxidation, a key biogeochemical process catalyzed by methanotrophic bacteria. Although aerobic methanotrophs showed remarkable resilience to a suite of physical disturbances induced as a single event, their resilience to recurring disturbances is less known. Here, using a rice field soil in a microcosm study, we determined whether recurrence and frequency of desiccation-rewetting impose an accumulating effect on the methanotrophic activity. The response of key aerobic methanotroph subgroups (type Ia, Ib, and II) were monitored using qPCR assays, and was supported by a t-RFLP analysis. The methanotrophic activity was resilient to recurring desiccation-rewetting, but increasing the frequency of the disturbance by twofold significantly decreased methane uptake rate. Both the qPCR and t-RFLP analyses were congruent, showing the dominance of type Ia/Ib methanotrophs prior to disturbance, and after disturbance, the recovering community was predominantly comprised of type Ia (Methylobacter) methanotrophs. Both type Ib and type II (Methylosinus/Methylocystis) methanotrophs were adversely affected by the disturbance, but type II methanotrophs showed recovery over time, indicating relatively higher resilience to the disturbance. This revealed distinct, yet unrecognized traits among the methanotroph community members. Our results show that recurring desiccation-rewetting before a recovery in community abundance had an accumulated effect, compromising methanotrophic activity. While methanotrophs may recover well following sporadic disturbances, their resilience may reach a 'tipping point' where activity no longer recovered if disturbance persists and increase in frequency.

  7. Sulfur metabolism in the extreme acidophile Acidithiobacillus caldus

    Directory of Open Access Journals (Sweden)

    Stefanie eMangold

    2011-02-01

    Full Text Available Given the challenges to life at low pH, an analysis of inorganic sulfur compound oxidation was initiated in the chemolithoautotrophic extremophile Acidithiobacillus caldus. A. caldus is able to metabolize elemental sulfur and a broad range of inorganic sulfur compounds. It has been implicated in the production of environmentally damaging acidic solutions as well as participating in industrial bioleaching operations where it forms part of microbial consortia used for the recovery of metal ions. Based upon the recently published A. caldus type strain genome sequence, a bioinformatic reconstruction of elemental sulfur and inorganic sulfur compound metabolism predicted genes included: sulfide quinone reductase (sqr, tetrathionate hydrolase (tth, two sox gene clusters potentially involved in thiosulfate oxidation (soxABXYZ, sulfur oxygenase reductase (sor, and various electron transport components. RNA transcript profiles by semi-quantitative reverse transcription PCR suggested up-regulation of sox genes in the presence of tetrathionate. Extensive gel based proteomic comparisons of total soluble and membrane enriched protein fractions during growth on elemental sulfur and tetrathionate identified differential protein levels from the two Sox clusters as well as several chaperone and stress proteins up-regulated in the presence of elemental sulfur. Proteomics results also suggested the involvement of heterodisulfide reductase (HdrABC in A. caldus inorganic sulfur compound metabolism. A putative new function of Hdr in acidophiles is discussed. Additional proteomic analysis evaluated protein expression differences between cells grown attached to solid, elemental sulfur versus planktonic cells. This study has provided insights into sulfur metabolism of this acidophilic chemolithotroph and gene expression during attachment to solid elemental sulfur.

  8. Methanotrophic bacteria in oilsands tailings ponds of northern Alberta

    Science.gov (United States)

    Saidi-Mehrabad, Alireza; He, Zhiguo; Tamas, Ivica; Sharp, Christine E; Brady, Allyson L; Rochman, Fauziah F; Bodrossy, Levente; Abell, Guy CJ; Penner, Tara; Dong, Xiaoli; Sensen, Christoph W; Dunfield, Peter F

    2013-01-01

    We investigated methanotrophic bacteria in slightly alkaline surface water (pH 7.4–8.7) of oilsands tailings ponds in Fort McMurray, Canada. These large lakes (up to 10 km2) contain water, silt, clay and residual hydrocarbons that are not recovered in oilsands mining. They are primarily anoxic and produce methane but have an aerobic surface layer. Aerobic methane oxidation was measured in the surface water at rates up to 152 nmol CH4 ml−1 water d−1. Microbial diversity was investigated via pyrotag sequencing of amplified 16S rRNA genes, as well as by analysis of methanotroph-specific pmoA genes using both pyrosequencing and microarray analysis. The predominantly detected methanotroph in surface waters at all sampling times was an uncultured species related to the gammaproteobacterial genus Methylocaldum, although a few other methanotrophs were also detected, including Methylomonas spp. Active species were identified via 13CH4 stable isotope probing (SIP) of DNA, combined with pyrotag sequencing and shotgun metagenomic sequencing of heavy 13C-DNA. The SIP-PCR results demonstrated that the Methylocaldum and Methylomonas spp. actively consumed methane in fresh tailings pond water. Metagenomic analysis of DNA from the heavy SIP fraction verified the PCR-based results and identified additional pmoA genes not detected via PCR. The metagenome indicated that the overall methylotrophic community possessed known pathways for formaldehyde oxidation, carbon fixation and detoxification of nitrogenous compounds but appeared to possess only particulate methane monooxygenase not soluble methane monooxygenase. PMID:23254511

  9. Recovery of methanotrophs from disturbance: population dynamics, evenness and functioning.

    Science.gov (United States)

    Ho, Adrian; Lüke, Claudia; Frenzel, Peter

    2011-04-01

    Biodiversity is claimed to be essential for ecosystem functioning, but is threatened by anthropogenic disturbances. Prokaryotes have been assumed to be functionally redundant and virtually inextinguishable. However, recent work indicates that microbes may well be sensitive to environmental disturbance. Focusing on methane-oxidizing bacteria as model organisms, we simulated disturbance-induced mortality by mixing native with sterilized paddy soil in two ratios, 1:4 and 1:40, representing moderate and severe die-offs. Disturbed microcosms were compared with an untreated control. Recovery of activity and populations was followed over 4 months by methane uptake measurements, pmoA-qPCR, pmoA-based terminal restriction fragment length polymorphism and a pmoA-based diagnostic microarray. Diversity and evenness of methanotrophs decreased in disturbed microcosms, but functioning was not compromised. We consistently observed distinctive temporal shifts between type I and type II methanotrophs, and a rapid population growth leading to even higher cell numbers comparing disturbed microcosms with the control. Overcompensating mortality suggested that population size in the control was limited by competition with other bacteria. Overall, methanotrophs showed a remarkable ability to compensate for die-offs.

  10. Feasibility of atmospheric methane removal using methanotrophic biotrickling filters

    Energy Technology Data Exchange (ETDEWEB)

    Yoon, Sukhwan; Carey, Jeffrey N.; Semrau, Jeremy D. [Michigan Univ., Ann Arbor, MI (United States). Dept. of Civil and Environmental Engineering

    2009-07-15

    Methane is a potent greenhouse gas with a global warming potential {proportional_to}23 times that of carbon dioxide. Here, we describe the modeling of a biotrickling filtration system composed of methane-consuming bacteria, i.e., methanotrophs, to assess the utility of these systems in removing methane from the atmosphere. Model results indicate that assuming the global average atmospheric concentration of methane, 1.7 ppmv, methane removal is ineffective using these methanotrophic biofilters as the methane concentration is too low to enable cell survival. If the concentration is increased to 500-6,000 ppmv, however, similar to that found above landfills and in concentrated animal feeding operations (factory farms), 4.98-35.7 tons of methane can be removed per biofilter per year assuming biotrickling filters of typical size (3.66 m in diameter and 11.5 m in height). Using reported ranges of capital, operational, and maintenance costs, the cost of the equivalent ton of CO{sub 2} removal using these systems is $90-$910 ($2,070-$20,900 per ton of methane), depending on the influent concentration of methane and if heating is required. The use of methanotrophic biofilters for controlling methane emissions is technically feasible and, provided that either the costs of biofilter construction and operation are reduced or the value of CO{sub 2} credits is increased, can also be economically attractive. (orig.)

  11. Leaching of pyrite by acidophilic heterotrophic iron-oxidizing bacteria in pure and mixed cultures

    Energy Technology Data Exchange (ETDEWEB)

    Bacelar-Nicolau, P.; Johnson, D.B. [Univ. of Wales, Bangor (United Kingdom). School of Biological Sciences

    1999-02-01

    Seven strains of heterotrophic iron-oxidizing acidophilic bacteria were examined to determine their abilities to promote oxidative dissolution of pyrite (FeS{sub 2}) when they were grown in pure cultures and in mixed cultures with sulfur-oxidizing Thiobacillus spp. Only one of the isolates (strain T-24) oxidized pyrite when it was grown in pyrite-basal salts medium. However, when pyrite-containing cultures were supplemented with 0.02% (wt/vol) yeast extract, most of the isolates oxidized pyrite, and one (strain T-24) promoted rates of mineral dissolution similar to the rates observed with the iron-oxidizing autotroph Thiobacillus ferroxidans. Pyrite oxidation by another isolate (strain T-21) occurred in cultures containing between 0.005 and 0.05% (wt/vol) yeast extract but was completely inhibited in cultures containing 0.5% yeast extract. Ferrous iron was also needed for mineral dissolution by the iron-oxidizing heterotrophs, indicating that these organisms oxidize pyrite via the indirect mechanism. Mixed cultures of three isolates (strains T-21, T-232, and T-24) and the sulfur-oxidizing autotroph Thiobacillus thiooxidans promoted pyrite dissolution; since neither strains T-21 and T-23 nor T. thiooxidans could oxidize this mineral in yeast extract-free media, this was a novel example of bacterial synergism. Mixed cultures of strains T-21 and T-23 and the sulfur-oxidizing mixotroph Thiobacillus acidophilus also oxidized pyrite but to a lesser extent than did mixed cultures containing T. thiooxidans. Pyrite leaching by strain T -23 grown in an organic compound-rich medium and incubated either shaken or unshaken was also assessed. The potential environmental significance of iron-oxidizing heterotrophs in accelerating pyrite oxidation is discussed.

  12. Identification of hopanoid, sterol, and tetrahymanol production in the aerobic methanotroph Methylomicrobium alcaliphilum 20Z

    Science.gov (United States)

    Welander, P. V.; Summons, R. E.

    2013-12-01

    Correlating the occurrence of molecular biosignatures preserved in the rock record with specific microbial taxa is a compelling strategy for studying microbial life in the context of the Earth's distant past. Polycyclic triterpenoids, including the hopanes and steranes, comprise classes of biomarkers that are readily detected in a variety of ancient sediments and are clearly recognized as the diagenetic products of modern day bacterial hopanoids and eukaryotic sterols. Thus, based on the distribution of these lipids in extant microbes, the occurrence of their diagenetic products in the rock record is often utilized as evidence for the existence of specific bacterial and eukaryotic taxa in ancient ecosystems. However, questions have arisen about our understanding of the taxonomic distribution of many of these molecular biomarkers in extant microbes. This is prompting reassessments of the use of polycyclic triterpenoids as geological proxies for microbial taxa, especially in the light of the poorly defined issue of microbial diversity. Recently, significant effort has been put forth to better understand the biosynthesis, function, and regulation of these lipid molecules in a variety of modern organisms so that a more informed interpretation of their occurrence in the rock record can be reached. Here we report the unprecedented production of three different classes of polycyclic triterpenoid biomarker lipids in one bacterium. Methylomicrobium alcaliphilum 20Z, a member of the Gammaproteobacteria, is a halotolerant alkaliphilic aerobic methanotroph previously isolated from a moderately saline soda lake in Tuva (Central Asia). In this study, M. alcaliphilum is shown to produce C-3 methylated and unmethylated aminohopanoids commonly associated with other mesophilic aerobic methanotrophs. In addition, this organism is also able to produce 4,4-dimethyl sterols and surprisingly, the gammacerane triterpenoid tetrahymanol. Previously, tetrahymanol production has only been

  13. [Leaching of copper ore of the Udokanskoe deposit at low temperatures by an association of acidophilic chemolithotrophic microorganisms].

    Science.gov (United States)

    Kondrat'eva, T F; Pivovarova, T A; Krylova, L N; Melamud, V S; Adamov, E V; Karavaĭko, G I

    2011-01-01

    Pure cultures of indigenous microorganisms Acidithiobacillus ferrooxidans strain TFUd, Leptospirillum ferrooxidans strain LUd, and Sulfobacillus thermotolerans strain SUd have been isolated from the oxidation zone of sulfide copper ore of the Udokanskoe deposit. Regimes of bacterial-chemical leaching of ore have been studied over a temperature range from -10 to +20 degrees C. Effects of pH, temperature, and the presence of microorganisms on the extraction of copper have been shown. Bacterial leaching has been detected only at positive values of temperature, and has been much more active at +20 than at +4 degrees C. The process of leaching was more active when the ore contained more hydrophilic and oxidized minerals. The possibility of copper ore leaching of the Udokanskoe deposit using sulfuric acid with pH 0.4 at negative values of temperature and applying acidophilic chemolithotrophic microorganisms at positive values of temperature and low pH values was shown.

  14. Sulphide bioxidation using native Acidithiobacillus ferrooxidans and thiooxidans, related acidophile strains from "El Zancudo" gold mine (Titiribí, Colombia

    Directory of Open Access Journals (Sweden)

    Diana M Ossa H

    2007-02-01

    Full Text Available This investigation was aimed at a laboratory evaluation of the action of native Acidithiobacillus ferrooxidans-and thiooxidans-related acidophile micro-organism (isolated from El Zancudo gold mine in Titiribí, Colombia on the oxidation of sulphides, mainly consisting of pyrite and arsenopyrite; sulphide concentration varied from 8% to 16% and particle size from 75 um to 106 um. pH and redox potential measurement revealed efficient oxidation for each culture over a 25-day period, reaching 1,4 pH and 600 mV redox potential. 20 g/L As and 60 g/L Fe levels were reached in solution. XRD, FTIR and SEM/EDX analysis revealed the presence of bacterial oxidation products such as jarosite and iron arsenates, as well as low and high corrosion in pyrites and arsenopyrites, respectively. Key words: chemolithotrophics, bioleaching, pyrite, arsenopyrite, applied mineralogy.

  15. [The use of real-time PCR technology to assess the effectiveness of methods of DNA extraction from cultures of acidophilic chemolithotrophic microorganisms].

    Science.gov (United States)

    Rogatykh, S V; Dokshukina, A A; Khaĭnasova, T S; Muradov, S V; Kofiadi, I A

    2011-01-01

    Comparative evaluation of efficiency of several methods of DNA extraction from storage cultures of acidophilic chemolithotrophic microorganism communities isolated from sulfide ores of Shanuch ore deposit (Kamchatka peninsula) was conducted. DNA extraction methods in various combinations of physical (heating to 65-98 degrees C, grinding with SiO2 particles), enzymatic (treatment with lysozyme and proteinase K), and chemical (GuSCN, CTAB and KOH) treatments were tested. The evaluation of efficiency was performed using Real-time PCR. The best result was obtained for the combined method based on GuSCN lysis activity (lysis at 65 degrees C) followed by purification with phenol and chloroform.

  16. TCE degradation in a methanotrophic attached-film bioreactor

    Energy Technology Data Exchange (ETDEWEB)

    Fennell, D.; Nelson, Y.M.; Underhill, S.E.; White, T.E.; Jewell, W.J. (Cornell Univ., Ithaca, NY (United States). Dept. of Agricultural and Biological Engineering)

    1993-09-20

    Trichloroethene was degraded in expanded-bed bioreactors operated with mixed-culture methanotrophic attached films. Biomass concentrations of 8 to 75 g volatile solids (VS) per liter static bed (L[sub sb]) were observed. Batch TCE degradation rates at 35C followed the Michaelis-Menten model, and a maximum TCE degradation rate (q[sub max]) of 10.6 mg TCE/gVS [center dot] day and a half velocity coefficient (K[sub s]) of 2.8 mg TCE/L were predicted. Continuous-flow kinetics also followed the Michaelis-Menten model, but other parameters may be limiting, such as dissolved copper and dissolved methane-q[sub max] and K[sub s] were 2.9 mg TCE/gVS [center dot] day and 1.5 mg TCE/L, respectively, at low copper concentrations (0.003 to 0.006 mg Cu/L). The maximum rates decreased substantially with small increases in dissolved copper. Methane consumption during continuous-flow operation varied from 23 to 1,200 g CH[sub 4]/g TCE degraded. Increasing the influent dissolved methane concentration from 0.01 mg/L to 5.4 mg/L reduced the TCE degradation rate by nearly an order of magnitude at 21C. Exposure of biofilms to 1.4 mg/L tetrachloroethene (PCE) at 35C resulted in the loss of methane utilization ability. Tests with methanotrophs grown on granular activated carbon indicated that lower effluent TCE concentrations could be obtained. The low efficiencies of TCE removal and low degradation rates obtained at 35C suggest that additional improvements will be necessary to make methanotrophic TCE treatment attractive.

  17. Genes and enzymes of ectoine biosynthesis in halotolerant methanotrophs.

    Science.gov (United States)

    Reshetnikov, Alexander S; Khmelenina, Valentina N; Mustakhimov, Ildar I; Trotsenko, Yuri A

    2011-01-01

    Ectoine (1,4,5,6-tetrahydro-2-methyl-4-pyrimidine carboxylic acid) is a widely distributed compatible solute accumulated by halophilic and halotolerant microorganisms to prevent osmotic stress in highly saline environments. Ectoine as a highly water keeping compound stabilizing biomolecules and whole cells can be used in scientific work, cosmetics, and medicine. Detailed understanding of the organization/regulation of the ectoine biosynthetic pathway in various producers is an active area of research. Here we review current knowledge on some genetic and enzymatic aspects of ectoine biosynthesis in halophilic and halotolerant methanotrophs. By using PCR methodology, the genes coding for the specific enzymes of ectoine biosynthesis, diaminobutyric acid (DABA) aminotransferase (EctB), DABA acetyltransferase (EctA), and ectoine synthase (EctC), were identified in several methanotrophic species. Organization of these genes in either ectABC or ectABC-ask operons, the latter additionally encoding aspartate kinase isozyme (Ask), correlated well with methanotroph halotolerance and intracellular ectoine level. A new gene, ectR1 encoding the MarR-like transcriptional regulatory protein EctR1, negatively controlling transcription of ectoine biosynthetic genes was found upstream of ectABC-ask operon in Methylomicrobium alcaliphilum 20Z. The ectR-like genes were also found in halotolerant methanol utilizers Methylophaga alcalica and Methylophaga thalassica as well as in several genomes of nonmethylotrophic species. The His(6)-tagged DABA acetyltransferases from Mm. alcaliphilum, M. alcalica, and M. thalassica were purified and the enzyme properties were found to correlate with the ecophysiologies of these bacteria. All these discoveries should be very helpful for better understanding the biosynthetic mechanism of this important natural compound, and for the targeted metabolic engineering of its producers.

  18. Spatial patterns of methane oxidation and methanotrophic diversity in landfill cover soils of southern China.

    Science.gov (United States)

    Chi, Zi-Fang; Lu, Wen-Jing; Wang, Hong-Tao

    2015-04-01

    Aerobic CH4 oxidation is an important CH4 sink in landfills. To investigate the distribution and community diversity of methanotrophs and link with soil characteristics and operational parameters (e.g., concentrations of O2, CH4), cover soil samples were collected at different locations and depths from the Mengzi semi-aerobic landfill (SAL) in Yunnan Province of southern China. Specific PCR followed by denaturing gradient gel electrophoresis and realtime PCR were used to examine methanotrophs in the landfill cover soils. The results showed that different locations did harbor distinct methanotroph communities. Methanotrophs were more abundant in areas near the venting pipes because of the higher O2 concentrations. The depth of 20-25 cm, where the ratio of the CH4 to O2 was within the range from 1.3 to 8.6, was more conducive to the growth of CH4-oxidizing bacteria. Type II methanotrophs dominated in all samples compared with Type I methanotrophs, as evidenced by the high ratio of Type II to Type I methanotrophic copy numbers (from 1.76 to 11.60). The total copy numbers of methanotrophs detected were similar to other ecosystems, although the CH4 concentration was much higher in SAL cover soil. Methylobacter and Methylocystis were the most abundant Type I and Type II methanotrophs genera, respectively, in the Mengzi SAL. The results suggested that SALs could provide a special environment with both high concentrations of CH4 and O2 for methanotrophs, especially around the vertical venting pipes.

  19. Detection of autotrophic verrucomicrobial methanotrophs in a geothermal environment using stable isotope probing

    Directory of Open Access Journals (Sweden)

    Christine eSharp

    2012-08-01

    Full Text Available Genomic analysis of the methanotrophic verrucomicrobium Methylacidiphilum infernorum strain V4 has shown that most pathways conferring its methanotrophic lifestyle are similar to those found in proteobacterial methanotrophs. However, due to the large sequence divergence of its methane monooxygenase-encoding genes (pmo, ‘universal’ pmoA polymerase chain reaction (PCR primers do not target these bacteria. Unlike proteobacterial methanotrophs, Methylacidiphilum fixes carbon autotrophically, and uses methane only for energy generation. As a result, techniques used to detect methanotrophs in the environment such as 13CH4-stable isotope probing (SIP and pmoA-targeted PCR do not detect verrucomicrobial methanotrophs, and they may have been overlooked in previous environmental studies. We developed a modified SIP technique to identify active methanotrophic verrucomicrobia in the environment by labelling with 13CO2 and 13CH4, individually and in combination. Testing the protocol in M. infernorum strain V4 resulted in assimilation of 13CO2 but not 13CH4, verifying its autotrophic lifestyle. To specifically detect methanotrophs (as opposed to other autotrophs via 13CO2-SIP, a quantitative PCR (qPCR assay specific for verrucomicrobial-pmoA genes was developed and used in combination with SIP. Incubation of an acidic, high-temperature geothermal soil with 13CH4 + 12CO2 caused little shift in the density distribution of verrucomicrobial-pmoA genes relative to controls. However, labelling with 13CO2 in combination with 12CH4 or 13CH4 induced a strong shift in the distribution of verrucomicrobial-pmoA genes towards the heavy DNA fractions. The modified SIP technique demonstrated that the primary methanotrophs active in the soil were autotrophs and belonged to the Verrucomicrobia. This is the first demonstration of autotrophic, non-proteobacterial methanotrophy in situ, and provides a tool to detect verrucomicrobial methanotrophs in other ecosystems.

  20. Soluble Methane Monooxygenase Production and Trichloroethylene Degradation by a Type I Methanotroph, Methylomonas methanica 68-1

    Science.gov (United States)

    Koh, Sung-Cheol; Bowman, John P.; Sayler, Gary S.

    1993-01-01

    A methanotroph (strain 68-1), originally isolated from a trichloroethylene (TCE)-contaminated aquifer, was identified as the type I methanotroph Methylomonas methanica on the basis of intracytoplasmic membrane ultrastructure, phospholipid fatty acid profile, and 16S rRNA signature probe hybridization. Strain 68-1 was found to oxidize naphthalene and TCE via a soluble methane monooxygenase (sMMO) and thus becomes the first type I methanotroph known to be able to produce this enzyme. The specific whole-cell sMMO activity of 68-1, as measured by the naphthalene oxidation assay and by TCE biodegradation, was comparatively higher than sMMO activity levels in Methylosinus trichosporium OB3b grown in the same copper-free conditions. The maximal naphthalene oxidation rates of Methylomonas methanica 68-1 and Methylosinus trichosporium OB3b were 551 ± 27 and 321 ± 16 nmol h-1 mg of protein -1, respectively. The maximal TCE degradation rates of Methylomonas methanica 68-1 and Methylosinus trichosporium OB3b were 2,325 ± 260 and 995 ± 160 nmol h-1 mg of protein-1, respectively. The substrate affinity of 68-1 sMMO to naphthalene (Km, 70 ± 4 μM) and TCE (Km, 225 ± 13 μM), however, was comparatively lower than that of the sMMO of OB3b, which had affinities of 40 ± 3 and 126 ± 8 μM, respectively. Genomic DNA slot and Southern blot analyses with an sMMO gene probe from Methylosinus trichosporium OB3b showed that the sMMO genes of 68-1 have little genetic homology to those of OB3b. This result may indicate the evolutionary diversification of the sMMOs. Images PMID:16348920

  1. Metal resistance or tolerance? Acidophiles confront high metal loads via both abiotic and biotic mechanisms

    Directory of Open Access Journals (Sweden)

    Mark eDopson

    2014-04-01

    Full Text Available All metals are toxic at high concentrations and consequently their intracellular concentrations must be regulated. Acidophilic microorganisms have an optimum growth pH < 3 and proliferate in natural and anthropogenic low pH environments. Some acidophiles are involved in the catalysis of sulfide mineral dissolution, resulting in high concentrations of metals in solution. Acidophiles are often described as highly metal resistant via mechanisms such as multiple and/or more efficient active resistance systems than are present in neutrophiles. However, this is not the case for all acidophiles and we contend that their growth in high metal concentrations is partially due to an intrinsic tolerance as a consequence of the environment in which they live. In this perspective, we highlight metal tolerance via complexation of free metals by sulfate ions and passive tolerance to metal influx via an internal positive cytoplasmic transmembrane potential. These tolerance mechanisms have been largely ignored in past studies of acidophile growth in the presence of metals and should be taken into account.

  2. Gene Loss and Horizontal Gene Transfer Contributed to the Genome Evolution of the Extreme Acidophile “Ferrovum”

    Science.gov (United States)

    Ullrich, Sophie R.; González, Carolina; Poehlein, Anja; Tischler, Judith S.; Daniel, Rolf; Schlömann, Michael; Holmes, David S.; Mühling, Martin

    2016-01-01

    Acid mine drainage (AMD), associated with active and abandoned mining sites, is a habitat for acidophilic microorganisms that gain energy from the oxidation of reduced sulfur compounds and ferrous iron and that thrive at pH below 4. Members of the recently proposed genus “Ferrovum” are the first acidophilic iron oxidizers to be described within the Betaproteobacteria. Although they have been detected as typical community members in AMD habitats worldwide, knowledge of their phylogenetic and metabolic diversity is scarce. Genomics approaches appear to be most promising in addressing this lacuna since isolation and cultivation of “Ferrovum” has proven to be extremely difficult and has so far only been successful for the designated type strain “Ferrovum myxofaciens” P3G. In this study, the genomes of two novel strains of “Ferrovum” (PN-J185 and Z-31) derived from water samples of a mine water treatment plant were sequenced. These genomes were compared with those of “Ferrovum” sp. JA12 that also originated from the mine water treatment plant, and of the type strain (P3G). Phylogenomic scrutiny suggests that the four strains represent three “Ferrovum” species that cluster in two groups (1 and 2). Comprehensive analysis of their predicted metabolic pathways revealed that these groups harbor characteristic metabolic profiles, notably with respect to motility, chemotaxis, nitrogen metabolism, biofilm formation and their potential strategies to cope with the acidic environment. For example, while the “F. myxofaciens” strains (group 1) appear to be motile and diazotrophic, the non-motile group 2 strains have the predicted potential to use a greater variety of fixed nitrogen sources. Furthermore, analysis of their genome synteny provides first insights into their genome evolution, suggesting that horizontal gene transfer and genome reduction in the group 2 strains by loss of genes encoding complete metabolic pathways or physiological features

  3. A novel acidophile community populating waste ore deposits at an acid mine drainage site

    Institute of Scientific and Technical Information of China (English)

    HAO Chun-bo; ZHANG Hong-xun; BAI Zhi-hui; HU Qing; ZHANG Bao-guo

    2007-01-01

    Waste ore samples (pH 3.0) were collected at an acid mine drainage site in Anhui, China. The present acidophilic microbial community in the waste ore was studied with 16S rRNA gene clone library and denaturing gradient gel electrophoresis (DGGE). Eighteen different clones were identified and affiliated with Actinobacteria, low G + C Gram-positives, Thermomicrobia, Acidobacteria, Proteobacteria, Candidate division TM7, and Planctomycetes. Phylogenetic analysis of 16S rRNA gene sequences revealed a diversity of acidophiles in the samples that were mostly novel. It is unexpected that the moderately thermophilic acidophiles were abundant in the acidic ecosystem and may play a great role in the generation of AMD. The result of DGGE was consistent with that of clone library analysis. These findings help in the better understanding of the generation mechanism of AMD and in developing a more efficient method to control AMD.

  4. The Ecology of Acidophilic Microorganisms in the Corroding Concrete Sewer Environment.

    Science.gov (United States)

    Li, Xuan; Kappler, Ulrike; Jiang, Guangming; Bond, Philip L

    2017-01-01

    Concrete corrosion is one of the most significant problems affecting valuable sewer infrastructure on a global scale. This problem occurs in the aerobic zone of the sewer, where a layer of surface corrosion develops on the exposed concrete and the surface pH is typically lowered from around 11-10 (pristine concrete) to pH 2-4. Acidophilic microorganisms become established as biofilms within the concrete corrosion layer and enhance the loss of concrete mass. Until recently, the acidophilic community was considered to comprise relatively few species of microorganisms, however, the biodiversity of the corrosion community is now recognized as being extensive and varying from different sewer environmental conditions. The diversity of acidophiles in the corrosion communities includes chemolithoautotrophs, chemolithoheterotrophs, and chemoorganoheterotrophs. The activity of these microorganisms is strongly affected by H2S levels in the sewer gas phase, although CO2, organic matter, and iron in the corrosion layer influence this acidic ecosystem. This paper briefly presents the conditions within the sewer that lead to the development of concrete corrosion in that environment. The review focuses on the acidophilic microorganisms detected in sewer corrosion environments, and then summarizes their proposed functions and physiology, especially in relation to the corrosion process. To our knowledge, this is the first review of acidophilic corrosion microbial communities, in which, the ecology and the environmental conditions (when available) are considered. Ecological studies of sewer corrosion are limited, however, where possible, we summarize the important metabolic functions of the different acidophilic species detected in sewer concrete corrosion layers. It is evident that microbial functions in the acidic sewer corrosion environment can be linked to those occurring in the analogous acidic environments of acid mine drainage and bioleaching.

  5. Genomics of the thermo-acidophilic red alga Galdieria sulphuraria

    Science.gov (United States)

    Barbier, Guillaume G.; Zimmermann, Marc; Weber, Andreas P. M.

    2005-09-01

    Extremophilic organisms dwell in environments that are characterized by high or low temperatures (thermophiles or psychrophiles), very low or high pH-values (acidophiles or alkalophiles), high salt concentrations (halophiles), high pressure (barophiles), or extreme drought (xerophiles). Many extremophiles are microbes, and many also belong to the prokaryota. Galdieria sulphuraria, however, is a member of a group of extremophilic eukaryotes that are named Cyanidiales. Cyanidiales are unicellular red micro-algae that occur worldwide in hot acidic waters, volcanic calderas, and in human-made acidic environments such as acidic mine drainage. G. sulphuraria has a unique position within the Cyanidiales because, in contrast to the other obligate photoautotrophic members of this group, it is able to grow photoautotrophically, mixotrophically, and heterotrophically. It is not only resistant to acid (pH 0) and heat (56oC), but also to high salt (1.5 M NaCl), toxic metals, and many other abiotic stressors. This unusual combination of features such as thermophily, acidophily, resistance to a wide array of abiotic stressors, and an extraordinary metabolic plasticity make G. sulphuraria highly interesting model organism to study adaptation to extreme environments. We have started a genomics approach to gain insight into the biology of G. sulphuraria and to identify genes and gene products critical for survival under extreme conditions. To this end, we pursue a whole-genome, shotgun sequencing approach towards unraveling the genome sequence of G. sulphuraria. We report here on the status quo of the genome-sequencing project and we summarize what we have learned to date from the genome sequence about the biology of this truly unique extremophile.

  6. Genome Analysis of the Biotechnologically Relevant Acidophilic Iron Oxidising Strain JA12 Indicates Phylogenetic and Metabolic Diversity within the Novel Genus "Ferrovum".

    Directory of Open Access Journals (Sweden)

    Sophie R Ullrich

    Full Text Available Members of the genus "Ferrovum" are ubiquitously distributed in acid mine drainage (AMD waters which are characterised by their high metal and sulfate loads. So far isolation and microbiological characterisation have only been successful for the designated type strain "Ferrovum myxofaciens" P3G. Thus, knowledge about physiological characteristics and the phylogeny of the genus "Ferrovum" is extremely scarce.In order to access the wider genetic pool of the genus "Ferrovum" we sequenced the genome of a "Ferrovum"-containing mixed culture and successfully assembled the almost complete genome sequence of the novel "Ferrovum" strain JA12.The genome-based phylogenetic analysis indicates that strain JA12 and the type strain represent two distinct "Ferrovum" species. "Ferrovum" strain JA12 is characterised by an unusually small genome in comparison to the type strain and other iron oxidising bacteria. The prediction of nutrient assimilation pathways suggests that "Ferrovum" strain JA12 maintains a chemolithoautotrophic lifestyle utilising carbon dioxide and bicarbonate, ammonium and urea, sulfate, phosphate and ferrous iron as carbon, nitrogen, sulfur, phosphorous and energy sources, respectively.The potential utilisation of urea by "Ferrovum" strain JA12 is moreover remarkable since it may furthermore represent a strategy among extreme acidophiles to cope with the acidic environment. Unlike other acidophilic chemolithoautotrophs "Ferrovum" strain JA12 exhibits a complete tricarboxylic acid cycle, a metabolic feature shared with the closer related neutrophilic iron oxidisers among the Betaproteobacteria including Sideroxydans lithotrophicus and Thiobacillus denitrificans. Furthermore, the absence of characteristic redox proteins involved in iron oxidation in the well-studied acidophiles Acidithiobacillus ferrooxidans (rusticyanin and Acidithiobacillus ferrivorans (iron oxidase indicates the existence of a modified pathway in "Ferrovum" strain JA12

  7. Photochemical Performance of the Acidophilic Red Alga Cyanidium sp. in a pH Gradient

    Science.gov (United States)

    Kvíderová, Jana

    2012-06-01

    The acidophilic red alga Cyanidium sp. is one of the dominant mat-forming species in the highly acidic waters of Río Tinto, Spain. The culture of Cyanidium sp., isolated from a microbial mat sample collected at Río Tinto, was exposed to 9 different pH conditions in a gradient from 0.5 to 5 for 24 h and its physiological status evaluated by variable chlorophyll a fluorescence kinetics measurements. Maximum quantum yield was determined after 30 min, 1 h, 2 h, 4 h, 6 h and 24 h of exposure after 15 min dark adaptation. The effect of pH on photochemical activity of Cyanidium sp. was observable as early as 30 min after exposure and the pattern remained stable or with only minor modifications for 24 h. The optimum pH ranged from 1.5 to 2.5. A steep decrease of the photochemical activity was observed at pH below 1 even after 30 min of exposure. Although the alga had tolerated the exposure to pH = 1 for at least 6 h, longer (24 h) exposure resulted in reduction of the photochemical activity. At pH above 2.5, the decline was more moderate and its negative effect on photochemistry was less severe. According to the fluorescence measurements, the red alga Cyanidium sp. is well-adapted to prevailing pH at its original locality at Río Tinto, i.e. pH of 1 to 3. The short-term survival in pH < 1.5 may be adaptation to rare exposures to such low pH in the field. The tolerance of pH above 3 could be caused by adaptation to the microenvironment of the inner parts of microbial mats in which Cyanidium sp. usually dominates and where higher pH could occur due to photosynthetic oxygen production.

  8. Acquisition of useful and high ability genes for acidophilic bacteria; Kosansei saikin ni takai noryoku wo fuyosuru idenshi no kakutoku

    Energy Technology Data Exchange (ETDEWEB)

    Senda, T.; Inoue, C.; Shinbori, Y. [Tohoku University, Sendai (Japan)

    1997-02-01

    This effort aims at the development of high-performance bacteria usable in bio-leaching in metal smelting by acquiring genes capable of realizing such. A method is used of choosing some isolated strains exhibiting high-performance traits and acquiring target genes therefrom by use of genetic engineering. Approximately 200 kinds in the aggregate of acidophilic bacteria are currently available for the study, including isolated iron-oxidizing and sulfur-oxidizing bacteria, standard species acquired for the study, and strains previously isolated by the laboratory. The bacteria are tested with respect to their Fe{sup 2+}-oxidizing rates, sulfur-oxidizing capabilities, and strength to withstand inhibiting substances (Ag{sup +}, Cl{sup -}, Mo{sup 6+}, etc.), which results in the nomination of 8 strains. The study planned to follow includes processes involving the extraction of chromosome DNAs from the 8 strains and their refinement, gene cloning by the Southern hybridization method, determination of their base sequences, determination of the difference between the strains in point of gene expression, and investigations of the relations that the results of these processes bear toward the said high-performance traits. Also under way is a study about the infuence-exerting factors revealed during the evaluation of the abilities of acidphlic bacteria. 2 refs., 2 tabs.

  9. Acidophilic methanogens and their applications in anaerobic digestion.%嗜酸产甲烷菌及其在厌氧处理中的应用

    Institute of Scientific and Technical Information of China (English)

    郭晓慧; 吴伟祥; 韩志英; 石德智

    2011-01-01

    产甲烷菌在自然界碳素循环过程中发挥着重要作用.酸性泥炭沼泽环境中存在着多种未知的产甲烷古菌,其中嗜酸产甲烷菌因其特殊的生长代谢特征近年来引起学者的广泛关注.若将嗜酸产甲烷菌应用于高浓度有机废物或废水的厌氧消化过程中,可从本质上克服因酸积累造成的产甲烷抑制,减少运行成本,扩展厌氧消化处理技术的应用范围.本文综述了嗜酸产甲烷菌的富集分离培养方法、生理生化特性、代谢特征及相关分子生物学研究等内容,并对其在厌氧处理中的应用前景进行了分析和展望,提出了未来研究的方向.%Methanogens play an important role in global carbon cycle. There exists a range of unknown methanogenic archaea in acidic peat lands. among which. acidophilic methanogens have attracted increasing research interests because of their special metabolic characteristics. To introduce acidophilic methanogens in the anaerobic digestion process of high concentration organic wastes or waste water could essentially overcome the inhibition of acid accumulation on the methanogens and help reduce the operation cost. broadening the industrial application of anaerobic bio-treatment technology. In this paper, we reviewed the recent researches on acidophilic methanogens, with the focus on enrichment and isolation methods, physiological and biochemical characters. metabolic characteristics. and application of molecular biology. The potential applications of acidophilic methanogens in anaerobic digestion process were analyzed and proposed, and the directions for further researches were suggested.

  10. Acidophilic green alga Pseudochlorella sp. YKT1 accumulates high amount of lipid droplets under a nitrogen-depleted condition at a low-pH.

    Directory of Open Access Journals (Sweden)

    Shunsuke Hirooka

    Full Text Available Microalgal storage lipids are considered to be a promising source for next-generation biofuel feedstock. However, microalgal biodiesel is not yet economically feasible due to the high cost of production. One of the reasons for this is that the use of a low-cost open pond system is currently limited because of the unavoidable contamination with undesirable organisms. Extremophiles have an advantage in culturing in an open pond system because they grow in extreme environments toxic to other organisms. In this study, we isolated the acidophilic green alga Pseudochlorella sp. YKT1 from sulfuric acid mine drainage in Nagano Prefecture, Japan. The vegetative cells of YKT1 display the morphological characteristics of Trebouxiophyceae and molecular phylogenetic analyses indicated it to be most closely related to Pseudochlorella pringsheimii. The optimal pH and temperature for the growth of YKT1 are pH 3.0-5.0 and a temperature 20-25°C, respectively. Further, YKT1 is able to grow at pH 2.0 and at 32°C, which corresponds to the usual water temperature in the outdoors in summer in many countries. YKT1 accumulates a large amount of storage lipids (∼30% of dry weigh under a nitrogen-depleted condition at low-pH (pH 3.0. These results show that acidophilic green algae will be useful for industrial applications by acidic open culture systems.

  11. Influence of light intensity on methanotrophic bacterial activity in Petit Saut Reservoir, French Guiana.

    Science.gov (United States)

    Dumestre, J F; Guézennec, J; Galy-Lacaux, C; Delmas, R; Richard, S; Labroue, L

    1999-02-01

    One year after impoundment in January 1994, methanotrophic bacteria in Petit Saut Reservoir (French Guiana) were active at the oxic-anoxic interface. This activity was revealed by the sudden extinction of diffusive methane emission (600 metric tons of CH4. day-1 for the whole lake surface area, i.e., 360 km2). Lifting of inhibition was suspected. After reviewing the potential inhibitors of this physiological guild (O2, NH4+, sulfides) and considering the similarities with nitrifiers, we suggest that sunlight influenced the methanotrophic bacteria. On the basis of phospholipid analysis, only a type II methanotrophic community was identified in the lake. Both growth and methanotrophic activity of an enriched culture, obtained in the laboratory, were largely inhibited by illumination over 150 microeinsteins. m-2. s-1. These results were confirmed on a pure culture of Methylosinus trichosporium OB3B. In situ conditions showed that water transparency was quite stable in 1994 and 1995 and that the oxycline moved steadily deeper until January 1995. Considering the mean illumination profile during this period, we showed that removal of methanotrophic growth inhibition could only occur below a 2-m depth. The oxycline reached this level in October 1994, allowing methanotrophic bacteria to develop and to consume the entire methane emission 4 months later.

  12. Heavy metal resistance strategies of acidophilic bacteria and their acquisition: importance for biomining and bioremediation.

    Science.gov (United States)

    Navarro, Claudio A; von Bernath, Diego; Jerez, Carlos A

    2013-01-01

    Microbial solubilizing of metals in acid environments is successfully used in industrial bioleaching of ores or biomining to extract metals such as copper, gold, uranium and others. This is done mainly by acidophilic and other microorganisms that mobilize metals and generate acid mine drainage or AMD, causing serious environmental problems. However, bioremediation or removal of the toxic metals from contaminated soils can be achieved by using the specific properties of the acidophilic microorganisms interacting with these elements. These bacteria resist high levels of metals by using a few "canonical" systems such as active efflux or trapping of the metal ions by metal chaperones. Nonetheless, gene duplications, the presence of genomic islands, the existence of additional mechanisms such as passive instruments for pH and cation homeostasis in acidophiles and an inorganic polyphosphate-driven metal resistance mechanism have also been proposed. Horizontal gene transfer in environmental microorganisms present in natural ecosystems is considered to be an important mechanism in their adaptive evolution. This process is carried out by different mobile genetic elements, including genomic islands (GI), which increase the adaptability and versatility of the microorganism. This mini-review also describes the possible role of GIs in metal resistance of some environmental microorganisms of importance in biomining and bioremediation of metal polluted environments such as Thiomonas arsenitoxydans, a moderate acidophilic microorganism, Acidithiobacillus caldus and Acidithiobacillus ferrooxidans strains ATCC 23270 and ATCC 53993, all extreme acidophiles able to tolerate exceptionally high levels of heavy metals. Some of these bacteria contain variable numbers of GIs, most of which code for high numbers of genes related to metal resistance. In some cases there is an apparent correlation between the number of metal resistance genes and the metal tolerance of each of these

  13. Assessing the Efficacy of the Aerobic Methanotrophic Biofilter in Methane Hydrate Environments

    Energy Technology Data Exchange (ETDEWEB)

    Valentine, David

    2012-09-30

    In October 2008 the University of California at Santa Barbara (UCSB) initiated investigations of water column methane oxidation in methane hydrate environments, through a project funded by the National Energy Technology Laboratory (NETL) entitled: assessing the efficacy of the aerobic methanotrophic biofilter in methane hydrate environments. This Final Report describes the scientific advances and discoveries made under this award as well as the importance of these discoveries in the broader context of the research area. Benthic microbial mats inhabit the sea floor in areas where reduced chemicals such as sulfide reach the more oxidizing water that overlies the sediment. We set out to investigate the role that methanotrophs play in such mats at locations where methane reaches the sea floor along with sulfide. Mats were sampled from several seep environments and multiple sets were grown in-situ at a hydrocarbon seep in the Santa Barbara Basin. Mats grown in-situ were returned to the laboratory and used to perform stable isotope probing experiments in which they were treated with 13C-enriched methane. The microbial community was analyzed, demonstrating that three or more microbial groups became enriched in methane?s carbon: methanotrophs that presumably utilize methane directly, methylotrophs that presumably consume methanol excreted by the methanotrophs, and sulfide oxidizers that presumably consume carbon dioxide released by the methanotrophs and methylotrophs. Methanotrophs reached high relative abundance in mats grown on methane, but other bacterial processes include sulfide oxidation appeared to dominate mats, indicating that methanotrophy is not a dominant process in sustaining these benthic mats, but rather a secondary function modulated by methane availability. Methane that escapes the sediment in the deep ocean typically dissolved into the overlying water where it is available to methanotrophic bacteria. We set out to better understand the efficacy of this

  14. Seasonal Variation in Abundance and Diversity of Bacterial Methanotrophs in Five Temperate Lakes

    Science.gov (United States)

    Samad, Md Sainur; Bertilsson, Stefan

    2017-01-01

    Lakes are significant sources of methane (CH4) to the atmosphere. Within these systems, methanotrophs consume CH4 and act as a potential biofilter mitigating the emission of this potent greenhouse gas. However, it is still not well understood how spatial and temporal variation in environmental parameters influence the abundance, diversity, and community structure of methanotrophs in lakes. To address this gap in knowledge, we collected water samples from three depths (surface, middle, and bottom) representing oxic to suboxic or anoxic zones of five different Swedish lakes in winter (ice-covered) and summer. Methanotroph abundance was determined by quantitative real time polymerase chain reaction and a comparison to environmental variables showed that temperature, season as well as depth, phosphate concentration, dissolved oxygen, and CH4 explained the observed variation in methanotroph abundance. Due to minimal differences in methane concentrations (0.19 and 0.29 μM for summer and winter, respectively), only a weak and even negative correlation was observed between CH4 and methanotrophs, which was possibly due to usage of CH4. Methanotrophs were present at concentrations ranging from 105 to 106 copies/l throughout the oxic (surface) and suboxic/anoxic (bottom) water mass of the lakes, but always contributed less than 1.3% to the total microbial community. Relative methanotroph abundance was significantly higher in winter than in summer and consistently increased with depth in the lakes. Phylogenetic analysis of pmoA genes in two clone libraries from two of the ice-covered lakes (Ekoln and Ramsen) separated the methanotrophs into five distinct clusters of Methylobacter sp. (Type I). Terminal restriction fragment length polymorphism analysis of the pmoA gene further revealed significant differences in methanotrophic communities between lakes as well as between winter and summer while there were no significant differences between water layers. The study provides new

  15. Multisite-specific archaeosine tRNA-guanine transglycosylase (ArcTGT) from Thermoplasma acidophilum, a thermo-acidophilic archaeon

    National Research Council Canada - National Science Library

    Kawamura, Takuya; Hirata, Akira; Ohno, Satoshi; Nomura, Yuichiro; Nagano, Tomoko; Nameki, Nobukazu; Yokogawa, Takashi; Hori, Hiroyuki

    2016-01-01

    ...) and the subsequent modification of preQ0 to G(+) by archaeosine synthase. However, tRNA(Leu) from Thermoplasma acidophilum, a thermo-acidophilic archaeon, exceptionally has two G(+)13 and G(+)15 modifications...

  16. Metabolites associated with adaptation of microorganisms to an acidophilic, metal-rich environment identified by stable-isotope-enabled metabolomics

    National Research Council Canada - National Science Library

    Mosier, Annika C; Justice, Nicholas B; Bowen, Benjamin P; Baran, Richard; Thomas, Brian C; Northen, Trent R; Banfield, Jillian F

    2013-01-01

    ... associated with the environmental matrix. We identified key metabolites associated with acidophilic and metal-tolerant microorganisms using stable isotope labeling coupled with untargeted, high-resolution mass spectrometry. We observed...

  17. Characterization of methanotrophic bacterial populations in natural and agricultural aerobic soils of the European Russia

    Science.gov (United States)

    Kravchenko, Irina; Sukhacheva, Marina; Kizilova, Anna

    2014-05-01

    Atmospheric methane contributes to about 20% of the total radiative forcing by long-lived greenhouse gases, and microbial methane oxidation in upland soils is the only biological sink of methane. Microbial methane oxidation in aerated upland soils is estimated as 15 - 45 Tg yr-1 or 3-9% of the annual sink. Therefore there is need of extensive research to characterize methanotrophic activity in various ecosystems for possible application to reduce atmospheric methane fluxes and to minimize global climate change. The vast majority of known aerobic methanotrophs belongs to the Proteobacteria and placed in the families Methylococcaceae in the Gammaproteobacteria, and Methylocystaceae and Beijerinckiaceae in the Alphaproteobacteria. Known exceptions include the phylum Verrucomicrobia and uncultured methanotrophs such as Candidatus 'Methylomirabilis oxyfera' affiliated with the 'NC10' phylum. Plenty of studies of aerobic methane oxidation and key players of the process have been performed on various types of soils, and it was found that Methylocystis spp and uncultivated methanotrophs are abundant in upland soils. Two of the uncultured groups are upland soil cluster alphaproteobacteria (USCa) and gammaproteobacteria (USCg), as revealed by cultivation-independent surveys of pmoA diversity. Russia is extremely rich in soil types due to its vast territories, and most of these soils have never been investigated from the aspect of methanotrophy. This study addresses methane oxidation activity and diversity of aerobic methanotrophic bacteria in eight types of natural aerobic soils, four of which also had been under agricultural use. Methane fluxes have been measured by in situ static chamber method and methane oxidation rates in soil samples - by radioisotope tracer (14CH4) technique. Changes in methanotroph diversity and abundance were assessed by cloning and Sanger sequencing, and quantitative real-time PCR of pmoA genes. Methanotrophic population of unmanaged soils turned

  18. Diversity and distribution of methanotrophic archaea at cold seeps.

    Science.gov (United States)

    Knittel, Katrin; Lösekann, Tina; Boetius, Antje; Kort, Renate; Amann, Rudolf

    2005-01-01

    In this study we investigated by using 16S rRNA-based methods the distribution and biomass of archaea in samples from (i) sediments above outcropping methane hydrate at Hydrate Ridge (Cascadia margin off Oregon) and (ii) massive microbial mats enclosing carbonate reefs (Crimea area, Black Sea). The archaeal diversity was low in both locations; there were only four (Hydrate Ridge) and five (Black Sea) different phylogenetic clusters of sequences, most of which belonged to the methanotrophic archaea (ANME). ANME group 2 (ANME-2) sequences were the most abundant and diverse sequences at Hydrate Ridge, whereas ANME-1 sequences dominated the Black Sea mats. Other seep-specific sequences belonged to the newly defined group ANME-3 (related to Methanococcoides spp.) and to the Crenarchaeota of marine benthic group B. Quantitative analysis of the samples by fluorescence in situ hybridization (FISH) showed that ANME-1 and ANME-2 co-occurred at the cold seep sites investigated. At Hydrate Ridge the surface sediments were dominated by aggregates consisting of ANME-2 and members of the Desulfosarcina-Desulfococcus branch (DSS) (ANME-2/DSS aggregates), which accounted for >90% of the total cell biomass. The numbers of ANME-1 cells increased strongly with depth; these cells accounted 1% of all single cells at the surface and more than 30% of all single cells (5% of the total cells) in 7- to 10-cm sediment horizons that were directly above layers of gas hydrate. In the Black Sea microbial mats ANME-1 accounted for about 50% of all cells. ANME-2/DSS aggregates occurred in microenvironments within the mat but accounted for only 1% of the total cells. FISH probes for the ANME-2a and ANME-2c subclusters were designed based on a comparative 16S rRNA analysis. In Hydrate Ridge sediments ANME-2a/DSS and ANME-2c/DSS aggregates differed significantly in morphology and abundance. The relative abundance values for these subgroups were remarkably different at Beggiatoa sites (80% ANME-2a, 20

  19. Diversity and Distribution of Methanotrophic Archaea at Cold Seeps†

    Science.gov (United States)

    Knittel, Katrin; Lösekann, Tina; Boetius, Antje; Kort, Renate; Amann, Rudolf

    2005-01-01

    In this study we investigated by using 16S rRNA-based methods the distribution and biomass of archaea in samples from (i) sediments above outcropping methane hydrate at Hydrate Ridge (Cascadia margin off Oregon) and (ii) massive microbial mats enclosing carbonate reefs (Crimea area, Black Sea). The archaeal diversity was low in both locations; there were only four (Hydrate Ridge) and five (Black Sea) different phylogenetic clusters of sequences, most of which belonged to the methanotrophic archaea (ANME). ANME group 2 (ANME-2) sequences were the most abundant and diverse sequences at Hydrate Ridge, whereas ANME-1 sequences dominated the Black Sea mats. Other seep-specific sequences belonged to the newly defined group ANME-3 (related to Methanococcoides spp.) and to the Crenarchaeota of marine benthic group B. Quantitative analysis of the samples by fluorescence in situ hybridization (FISH) showed that ANME-1 and ANME-2 co-occurred at the cold seep sites investigated. At Hydrate Ridge the surface sediments were dominated by aggregates consisting of ANME-2 and members of the Desulfosarcina-Desulfococcus branch (DSS) (ANME-2/DSS aggregates), which accounted for >90% of the total cell biomass. The numbers of ANME-1 cells increased strongly with depth; these cells accounted 1% of all single cells at the surface and more than 30% of all single cells (5% of the total cells) in 7- to 10-cm sediment horizons that were directly above layers of gas hydrate. In the Black Sea microbial mats ANME-1 accounted for about 50% of all cells. ANME-2/DSS aggregates occurred in microenvironments within the mat but accounted for only 1% of the total cells. FISH probes for the ANME-2a and ANME-2c subclusters were designed based on a comparative 16S rRNA analysis. In Hydrate Ridge sediments ANME-2a/DSS and ANME-2c/DSS aggregates differed significantly in morphology and abundance. The relative abundance values for these subgroups were remarkably different at Beggiatoa sites (80% ANME-2a, 20

  20. Sucrose metabolism in halotolerant methanotroph Methylomicrobium alcaliphilum 20Z.

    Science.gov (United States)

    But, Sergey Y; Khmelenina, Valentina N; Reshetnikov, Alexander S; Mustakhimov, Ildar I; Kalyuzhnaya, Marina G; Trotsenko, Yuri A

    2015-04-01

    Sucrose accumulation has been observed in some methylotrophic bacteria utilizing methane, methanol, or methylated amines as a carbon and energy source. In this work, we have investigated the biochemical pathways for sucrose metabolism in the model halotolerant methanotroph Methylomicrobium alcaliphilum 20Z. The genes encoding sucrose-phosphate synthase (Sps), sucrose-phosphate phosphatase (Spp), fructokinase (FruK), and amylosucrase (Ams) were co-transcribed and displayed similar expression levels. Functional Spp and Ams were purified after heterologous expression in Escherichia coli. Recombinant Spp exhibited high affinity for sucrose-6-phosphate and stayed active at very high levels of sucrose (K i  = 1.0 ± 0.6 M). The recombinant amylosucrase obeyed the classical Michaelis-Menten kinetics in the reactions of sucrose hydrolysis and transglycosylation. As a result, the complete metabolic network for sucrose biosynthesis and re-utilization in the non-phototrophic organism was reconstructed for the first time. Comparative genomic studies revealed analogous gene clusters in various Proteobacteria, thus indicating that the ability to produce and metabolize sucrose is widespread among prokaryotes.

  1. Synthesis of silver nanoparticles from two acidophilic strains of Pilimelia columellifera subsp. pallida and their antibacterial activities.

    Science.gov (United States)

    Golińska, Patrycja; Wypij, Magdalena; Rathod, Dnyaneshwar; Tikar, Sagar; Dahm, Hanna; Rai, Mahendra

    2016-05-01

    Biosynthesis of silver nanoparticles (AgNPs) is an eco-friendly approach by using different biological sources; for example, plants and microorganisms such as bacteria, fungi, and actinobacteria. In this report, we present the biological synthesis of silver nanoparticles (AgNPs) by acidophilic actinomycetes SL19 and SL24 strains isolated from pine forest soil (pH tracking analysis (NTA), Zeta potential, Fourier transform infrared spectroscopy (FTIR) and Transmission electron microscopy (TEM). FTIR analysis revealed the presence of proteins as a capping agent. TEM analysis confirmed the formation of spherical and polydispersed NPs of 12.7 and 15.9 nm sizes. The in vitro antibacterial activity of AgNPs alone and in combination with antibiotics was evaluated against clinical bacteria viz., Staphylococcus aureus, Bacillus subtilis, Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeruginosa, and uropathogens such as Enterobacter, S. aureus, P. aeruginosa, K. pneumoniae, and E. coli. The lowest MIC (40 μg ml(-1) ) was demonstrated by AgNPs synthesized from SL24 against E. coli. However, the AgNPs of SL19 showed lowest MIC (70 μg ml(-1) ) against S. aureus. The activity of antibiotic was enhanced, when tested in combination with silver nanoparticles synthesized from both actinobacterial strains.

  2. Characterization of a novel thiosulfate dehydrogenase from a marine acidophilic sulfur-oxidizing bacterium, Acidithiobacillus thiooxidans strain SH.

    Science.gov (United States)

    Sharmin, Sultana; Yoshino, Eriko; Kanao, Tadayoshi; Kamimura, Kazuo

    2016-01-01

    A marine acidophilic sulfur-oxidizing bacterium, Acidithiobacillus thiooxidans strain SH, was isolated to develop a bioleaching process for NaCl-containing sulfide minerals. Because the sulfur moiety of sulfide minerals is metabolized to sulfate via thiosulfate as an intermediate, we purified and characterized the thiosulfate dehydrogenase (TSD) from strain SH. The enzyme had an apparent molecular mass of 44 kDa and was purified 71-fold from the solubilized membrane fraction. Tetrathionate was the product of the TSD-oxidized thiosulfate and ferricyanide or ubiquinone was the electron acceptor. Maximum enzyme activity was observed at pH 4.0, 40 °C, and 200 mM NaCl. To our knowledge, this is the first report of NaCl-stimulated TSD activity. TSD was structurally different from the previously reported thiosulfate-oxidizing enzymes. In addition, TSD activity was strongly inhibited by 2-heptyl-4-hydroxy-quinoline N-oxide, suggesting that the TSD is a novel thiosulfate:quinone reductase.

  3. Methane source strength and energy flow shape methanotrophic communities in oxygen-methane counter-gradients.

    Science.gov (United States)

    Krause, Sascha; Lüke, Claudia; Frenzel, Peter

    2012-04-01

    The role of microbial diversity for ecosystem functioning has become an important subject in microbial ecology. Recent work indicates that microbial communities and microbial processes can be very sensitive to anthropogenic disturbances. However, to what extent microbial communities may change upon, resist to, or overcome disturbances might differ depending on substrate availability. We used soil from an Italian rice field in gradient microcosms, and analysed the response of methanotrophic communities to an NH4 (+) pulse as a potential disturbance under two different CH4 source strengths. We found a significant influence of source strength, i.e. the energy flow through the methanotrophic community, while NH4 (+) had no effect. Our data suggest that historical contingencies, i.e. nitrogen fertilization, led to an ammonium-tolerant MOB community. Methanotrophs were able to oxidize virtually all CH4 diffusing into the oxic-anoxic boundary layer regardless of NH4 (+) addition. Total and active methanotrophic communities were assessed by a pmoA-specific microarray. From the reservoir of dormant methanotrophs, different species became active with Methylobacter and an environmental cluster affiliated with paddy soils being indicative for high CH4 source strength. Thus, a microbial seed bank is an important prerequisite to maintain functioning in a fluctuating environment.

  4. Succession of methanotrophs in oxygen-methane counter-gradients of flooded rice paddies.

    Science.gov (United States)

    Krause, Sascha; Lüke, Claudia; Frenzel, Peter

    2010-12-01

    Little is known about population dynamics and contribution of specific taxa to methane oxidation in flooded rice paddies. In this article we investigate the succession of methanotrophs in oxygen-methane counter-gradients. We used a gradient microcosm system that simulates oxic-anoxic interfaces of a water-saturated paddy soils, and measured pmoA-based (gene encoding particulate methane monooxygenase) terminal restriction fragment length polymorphism (T-RFLP) profiles at both the transcription (mRNA) and the population (DNA) levels. The DNA T-RFLP profiles indicated that the methanotrophic community present clearly differed from the active methanotrophic community. We observed a succession of the methanotrophic community over time without any direct effect of pore water chemistry on the community structure. Both the total population and the active subpopulation changed with time, whereas methane oxidation rates remained nearly constant. Hence, we suggest that a diverse microbial seed bank of methanotrophs is important in maintaining the function in a dynamic ecosystem.

  5. Heavy metal bioleaching and sludge stabilization in a single-stage reactor using indigenous acidophilic heterotrophs.

    Science.gov (United States)

    Mehrotra, Akanksha; Sreekrishnan, T R

    2017-01-10

    Simultaneous sludge digestion and metal leaching (SSDML) have been reported at mesophilic temperature. It is generally perceived that while sludge stabilization is effected by heterotrophs at neutral pH, metal bioleaching is done by acidophilic autotrophs. However, little information is available on the microbial communities involved in the process. This study carried out SSDML in a single-stage reactor using sludge indigenous microorganisms and looked at the bacterial communities responsible for the process. Volatile suspended solids were reduced by more than 40%. The concentration of zinc, copper, chromium, cadmium and nickel decreased by more than 45% in the dry sludge. Acidophilic species of Alicyclobacillus genus were the dominant heterotrophs. A few heterotrophic bacteria were detected which can oxidize iron (Alicyclobacillus ferrooxydans, Alicyclobacillus ferripilum and Ferrimicrobium acidiphilum). Acidithiobacillus ferrooxidans (autotroph) was responsible for the oxidation of both iron and sulfur which lead to a change in the pH from neutral to acidic. The presence of acidophilic heterotrophs, which can oxidize either iron or sulfur, enhanced the efficiency of SSDML process with respect to sludge stabilization and metal leaching. This study shows that it is possible to carry out the SSDML in a single-stage reactor with indigenous microorganisms.

  6. Lessons from the genomes of extremely acidophilic bacteria and archaea with special emphasis on bioleaching microorganisms.

    Science.gov (United States)

    Cárdenas, Juan Pablo; Valdés, Jorge; Quatrini, Raquel; Duarte, Francisco; Holmes, David S

    2010-10-01

    This mini-review describes the current status of recent genome sequencing projects of extremely acidophilic microorganisms and highlights the most current scientific advances emerging from their analysis. There are now at least 56 draft or completely sequenced genomes of acidophiles including 30 bacteria and 26 archaea. There are also complete sequences for 38 plasmids, 29 viruses, and additional DNA sequence information of acidic environments is available from eight metagenomic projects. A special focus is provided on the genomics of acidophiles from industrial bioleaching operations. It is shown how this initial information provides a rich intellectual resource for microbiologists that has potential to open innovative and efficient research avenues. Examples presented illustrate the use of genomic information to construct preliminary models of metabolism of individual microorganisms. Most importantly, access to multiple genomes allows the prediction of metabolic and genetic interactions between members of the bioleaching microbial community (ecophysiology) and the investigation of major evolutionary trends that shape genome architecture and evolution. Despite these promising beginnings, a major conclusion is that the genome projects help focus attention on the tremendous effort still required to understand the biological principles that support life in extremely acidic environments, including those that might allow engineers to take appropriate action designed to improve the efficiency and rate of bioleaching and to protect the environment.

  7. Culture-independent detection of "TM7" bacteria in a streptomycin-resistant acidophilic nitrifying process

    Science.gov (United States)

    Kurogi, T.; Linh, N. T. T.; Kuroki, T.; Yamada, T.; Hiraishi, A.

    2014-02-01

    Nitrification in biological wastewater treatment processes has been believed for long time to take place under neutral conditions and is inhibited under acidic conditions. However, we previously constructed acidophilic nitrifying sequencing-batch reactors (ANSBRs) being capable of nitrification at level, this study was undertaken to construct streptomycin-resistant acidophilic nitrifying (SRAN) reactors and to demonstrate whether TM7 bacteria are abundant in these reactors. The SRAN reactors were constructed by seeding with nitrifying sludge from an ANSBR and cultivating with ammonium-containing mineral medium (pH 4.0), to which streptomycin at a concentration of 10, 30 and 50 mg L-1 was added. In all reactors, the pH varied between 2.7 and 4.0, and ammonium was completely converted to nitrate in every batch cycle. PCR-aided denaturing gradient gel electrophoresis (DGGE) targeting 16S rRNA genes revealed that some major clones assigned to TM7 bacteria and Gammaproteobacteria were constantly present during the overall period of operation. Fluorescence in situ hybridization (FISH) with specific oligonucleotide probes also showed that TM7 bacteria predominated in all SRAN reactors, accounting for 58% of the total bacterial population on average. Although the biological significance of the TM7 bacteria in the SRAN reactors are unknown, our results suggest that these bacteria are possibly streptomycin-resistant and play some important roles in the acidophilic nitrifying process.

  8. Fly ash application in nutrient poor agriculture soils: impact on methanotrophs population dynamics and paddy yields.

    Science.gov (United States)

    Singh, Jay Shankar; Pandey, Vimal Chandra

    2013-03-01

    There are reports that the application of fly ash, compost and press mud or a combination thereof, improves plant growth, soil microbial communities etc. Also, fly ash in combination with farmyard manure or other organic amendments improves soil physico-chemical characteristics, rice yield and microbial processes in paddy fields. However, the knowledge about the impact of fly ash inputs alone or in combination with other organic amendments on soil methanotrophs number in paddy soils is almost lacking. We hypothesized that fly ash application at lower doses in paddy agriculture soil could be a potential amendment to elevate the paddy yields and methanotrophs number. Here we demonstrate the impact of fly ash and press mud inputs on number of methanotrophs, antioxidants, antioxidative enzymatic activities and paddy yields at agriculture farm. The impact of amendments was significant for methanotrophs number, heavy metal concentration, antioxidant contents, antioxidant enzymatic activities and paddy yields. A negative correlation was existed between higher doses of fly ash-treatments and methanotrophs number (R(2)=0.833). The content of antioxidants and enzymatic activities in leaves of higher doses fly ash-treated rice plants increased in response to stresses due to heavy metal toxicity, which was negatively correlated with rice grain yield (R(2)=0.944) and paddy straw yield (R(2)=0.934). A positive correlation was noted between heavy metals concentrations and different antioxidant and enzymatic activities across different fly ash treated plots.The data of this study indicate that heavy metal toxicity of fly ash may cause oxidative stress in the paddy crop and the antioxidants and related enzymes could play a defensive role against phytotoxic damages. We concluded that fly ash at lower doses with press mud seems to offer the potential amendments to improving soil methanotrophs population and paddy crop yields for the nutrient poor agriculture soils.

  9. Nutrient amendments in soil DNA stable isotope probing experiments reduce the observed methanotroph diversity.

    Science.gov (United States)

    Cébron, Aurélie; Bodrossy, Levente; Stralis-Pavese, Nancy; Singer, Andrew C; Thompson, Ian P; Prosser, James I; Murrell, J Colin

    2007-02-01

    Stable isotope probing (SIP) can be used to analyze the active bacterial populations involved in a process by incorporating 13C-labeled substrate into cellular components such as DNA. Relatively long incubation times are often used with laboratory microcosms in order to incorporate sufficient 13C into the DNA of the target organisms. Addition of nutrients can be used to accelerate the processes. However, unnatural concentrations of nutrients may artificially change bacterial diversity and activity. In this study, methanotroph activity and diversity in soil was examined during the consumption of 13CH4 with three DNA-SIP experiments, using microcosms with natural field soil water conditions, the addition of water, and the addition of mineral salts solution. Methanotroph population diversity was studied by targeting 16S rRNA and pmoA genes. Clone library analyses, denaturing gradient gel electrophoresis fingerprinting, and pmoA microarray hybridization analyses were carried out. Most methanotroph diversity (type I and type II methanotrophs) was observed in non-amended SIP microcosms. Although this treatment probably best reflected the in situ environmental conditions, one major disadvantage of this incubation was that the incorporation of 13CH4 was slow and some cross-feeding of 13C occurred, thereby leading to labeling of nonmethanotroph microorganisms. Conversely, microcosms supplemented with mineral salts medium exhibited rapid consumption of 13CH4, resulting in the labeling of a less diverse population of only type I methanotrophs. DNA-SIP incubations using water-amended microcosms yielded faster incorporation of 13C into active methanotrophs while avoiding the cross-feeding of 13C.

  10. Shifts in identity and activity of methanotrophs in arctic lake sediments in response to temperature changes

    Science.gov (United States)

    He, Ruo; Wooller, Matthew J.; Pohlman, John W.; Quensen, John; Tiedje, James M.; Leigh, Mary Beth

    2012-01-01

    Methane (CH4) flux to the atmosphere is mitigated via microbial CH4 oxidation in sediments and water. As arctic temperaturesincrease, understanding the effects of temperature on the activity and identity of methanotrophs in arctic lake sediments is importantto predicting future CH4 emissions. We used DNA-based stable-isotope probing (SIP), quantitative PCR (Q-PCR), andpyrosequencing analyses to identify and characterize methanotrophic communities active at a range of temperatures (4°C, 10°C,and 21°C) in sediments (to a depth of 25 cm) sampled from Lake Qalluuraq on the North Slope of Alaska. CH4 oxidation activitywas measured in microcosm incubations containing sediments at all temperatures, with the highest CH4 oxidation potential of37.5 mol g1 day1 in the uppermost (depth, 0 to 1 cm) sediment at 21°C after 2 to 5 days of incubation. Q-PCR of pmoA and ofthe 16S rRNA genes of type I and type II methanotrophs, and pyrosequencing of 16S rRNA genes in 13C-labeled DNA obtained bySIP demonstrated that the type I methanotrophs Methylobacter, Methylomonas, and Methylosoma dominated carbon acquisitionfrom CH4 in the sediments. The identity and relative abundance of active methanotrophs differed with the incubation temperature.Methylotrophs were also abundant in the microbial community that derived carbon from CH4, especially in the deeper sediments(depth, 15 to 20 cm) at low temperatures (4°C and 10°C), and showed a good linear relationship (R0.82) with the relativeabundances of methanotrophs in pyrosequencing reads. This study describes for the first time how methanotrophiccommunities in arctic lake sediments respond to temperature variations.

  11. Effect of earthworms on the community structure of active methanotrophic bacteria in a landfill cover soil.

    Science.gov (United States)

    Héry, Marina; Singer, Andrew C; Kumaresan, Deepak; Bodrossy, Levente; Stralis-Pavese, Nancy; Prosser, Jim I; Thompson, Ian P; Murrell, J Colin

    2008-01-01

    In the United Kingdom, landfills are the primary anthropogenic source of methane emissions. Methanotrophic bacteria present in landfill biocovers can significantly reduce methane emissions via their capacity to oxidize up to 100% of the methane produced. Several biotic and abiotic parameters regulate methane oxidation in soil, such as oxygen, moisture, methane concentration and temperature. Earthworm-mediated bioturbation has been linked to an increase in methanotrophy in a landfill biocover soil (AC Singer et al., unpublished), but the mechanism of this trophic interaction remains unclear. The aims of this study were to determine the composition of the active methanotroph community and to investigate the interactions between earthworms and bacteria in this landfill biocover soil where the methane oxidation activity was significantly increased by the earthworms. Soil microcosms were incubated with 13C-CH4 and with or without earthworms. DNA and RNA were extracted to characterize the soil bacterial communities, with a particular emphasis on methanotroph populations, using phylogenetic (16S ribosomal RNA) and functional methane monooxygenase (pmoA and mmoX) gene probes, coupled with denaturing gradient-gel electrophoresis, clone libraries and pmoA microarray analyses. Stable isotope probing (SIP) using 13C-CH4 substrate allowed us to link microbial function with identity of bacteria via selective recovery of 'heavy' 13C-labelled DNA or RNA and to assess the effect of earthworms on the active methanotroph populations. Both types I and II methanotrophs actively oxidized methane in the landfill soil studied. Results suggested that the earthworm-mediated increase in methane oxidation rate in the landfill soil was more likely to be due to the stimulation of bacterial growth or activity than to substantial shifts in the methanotroph community structure. A Bacteroidetes-related bacterium was identified only in the active bacterial community of earthworm-incubated soil but

  12. Soluble cytochromes from the marine methanotroph Methylomonas sp. strain A4.

    OpenAIRE

    DiSpirito, A A; Lipscomb, J. D.; Lidstrom, M E

    1990-01-01

    Soluble c-type cytochromes are central to metabolism of C1 compounds in methylotrophic bacteria. In order to characterize the role of c-type cytochromes in methane-utilizing bacteria (methanotrophs), we have purified four different cytochromes, cytochromes c-554, c-553, c-552, and c-551, from the marine methanotroph Methylomonas sp. strain A4. The two major species, cytochromes c-554 and c-552, were monoheme cytochromes and accounted for 57 and 26%, respectively, of the soluble c-heme. The ap...

  13. Genome Analysis of the Biotechnologically Relevant Acidophilic Iron Oxidising Strain JA12 Indicates Phylogenetic and Metabolic Diversity within the Novel Genus “Ferrovum”

    Science.gov (United States)

    Ullrich, Sophie R.; Poehlein, Anja; Tischler, Judith S.; González, Carolina; Ossandon, Francisco J.; Daniel, Rolf; Holmes, David S.; Schlömann, Michael; Mühling, Martin

    2016-01-01

    Background Members of the genus “Ferrovum” are ubiquitously distributed in acid mine drainage (AMD) waters which are characterised by their high metal and sulfate loads. So far isolation and microbiological characterisation have only been successful for the designated type strain “Ferrovum myxofaciens” P3G. Thus, knowledge about physiological characteristics and the phylogeny of the genus “Ferrovum” is extremely scarce. Objective In order to access the wider genetic pool of the genus “Ferrovum” we sequenced the genome of a “Ferrovum”-containing mixed culture and successfully assembled the almost complete genome sequence of the novel “Ferrovum” strain JA12. Phylogeny and Lifestyle The genome-based phylogenetic analysis indicates that strain JA12 and the type strain represent two distinct “Ferrovum” species. “Ferrovum” strain JA12 is characterised by an unusually small genome in comparison to the type strain and other iron oxidising bacteria. The prediction of nutrient assimilation pathways suggests that “Ferrovum” strain JA12 maintains a chemolithoautotrophic lifestyle utilising carbon dioxide and bicarbonate, ammonium and urea, sulfate, phosphate and ferrous iron as carbon, nitrogen, sulfur, phosphorous and energy sources, respectively. Unique Metabolic Features The potential utilisation of urea by “Ferrovum” strain JA12 is moreover remarkable since it may furthermore represent a strategy among extreme acidophiles to cope with the acidic environment. Unlike other acidophilic chemolithoautotrophs “Ferrovum” strain JA12 exhibits a complete tricarboxylic acid cycle, a metabolic feature shared with the closer related neutrophilic iron oxidisers among the Betaproteobacteria including Sideroxydans lithotrophicus and Thiobacillus denitrificans. Furthermore, the absence of characteristic redox proteins involved in iron oxidation in the well-studied acidophiles Acidithiobacillus ferrooxidans (rusticyanin) and Acidithiobacillus

  14. Familial relationships in hyperthermo- and acidophilic archaeal viruses

    DEFF Research Database (Denmark)

    Happonen, Lotta Johanna; Redder, Peter; Peng, Xu;

    2010-01-01

    Archaea often live in extreme, harsh environments such as acidic hot springs and hypersaline waters. To date, only two icosahedrally symmetric, membrane-containing archaeal viruses, SH1 and Sulfolobus turreted icosahedral virus (STIV), have been described in detail. We report the sequence and three......-dimensional structure of a third such virus isolated from a hyperthermoacidophilic crenarchaeon, Sulfolobus strain G4ST-2. Characterization of this new isolate revealed it to be similar to STIV on the levels of genome and structural organization. The genome organization indicates that these two viruses have diverged...... from a common ancestor. Interestingly, the prominent surface turrets of the two viruses are strikingly different. By sequencing and mass spectrometry, we mapped several large insertions and deletions in the known structural proteins that could account for these differences and showed that both viruses...

  15. Group-specific quantification of methanotrophs in landfill gas-purged laboratory biofilters by tyramide signal amplification-fluorescence in situ hybridization.

    Science.gov (United States)

    Wang, Hong; Einola, Juha; Heinonen, Mirja; Kulomaa, Markku; Rintala, Jukka

    2008-09-01

    The aim of this study was to quantitatively analyse methanotrophs in two laboratory landfill biofilters at different biofilter depths and at temperatures which mimicked the boreal climatic conditions. Both biofilters were dominated by type I methanotrophs. The biofilter depth profiles showed that type I methanotrophs occurred in the upper layer, where relatively high O(2) and low CH(4) concentrations were present, whereas type II methanotrophs were mostly distributed in the zone with high CH(4) and low O(2) concentrations. The number of type I methanotrophic cells declined when the temperature was raised from 15 degrees C to 23 degrees C, but increased when lowered to 5 degrees C. A slight decrease in type II methanotrophs was also observed when the temperature was raised from 15 degrees C to 23 degrees C, whereas cell numbers remained constant when lowered to 5 degrees C. The results indicated that low temperature conditions favored both type I and type II methanotrophs in the biofilters.

  16. Methanotrophic production of copolymer, poly(hydroxybutyrate-co-hydroxyvalerate), with high hydroxyvalerate content

    Science.gov (United States)

    Type II methanotrophic bacteria are a promising production platform for PHA biopolymers. These bacteria are known to produce pure poly-3-hydroxybutyrate homopolymer. We describe the production of a wide range of PHB-co-HV co-polymers by the co-feeding of methane and valerate. The ratio of HB to HV m...

  17. Intracellular PHB conversion in a type II methanotroph studied by 13 C NMR

    NARCIS (Netherlands)

    Vecherskaya, M.; Dijkema, C.; Stams, A.J.M.

    2001-01-01

    Poly-g-hydroxybutyrate (PHB) formation under aerobic conditions via incorporation of [13C-2]acetate as a cosubstrate and its intracellular degradation under anaerobic conditions in a Type II methanotroph was studied by 13C NMR. During PHB synthesis in the presence of labelled acetate, low levels of

  18. Aerobic methanotrophic communities at the Red Sea brine-seawater interface

    KAUST Repository

    Abdallah, Rehab Z.

    2014-09-23

    Abdallah RZ, Adel M, Ouf A, Sayed A, Ghazy MA, Alam I, Essack M, Lafi FF, Bajic VB, El-Dorry H and Siam R (2014) Aerobic methanotrophic communities at the Red Sea brine-seawater interface. Front. Microbiol. 5:487. doi: 10.3389/fmicb.2014.00487

  19. Methanotrophic community dynamics in a seasonally anoxic fjord: Saanich Inlet, British Columbia

    Directory of Open Access Journals (Sweden)

    Mónica Torres-Beltrán

    2016-12-01

    Full Text Available Marine oxygen minimum zones (OMZs play disproportionate roles in nutrient and climate active trace gas cycling including nitrous oxide and methane, in the ocean. OMZs are currently expanding due to climate change making it increasingly important to identify microbial controls on trace gas cycling at the individual, population and community levels. Here we present a two-year survey of the microbial community along seasonal redox gradients in Saanich Inlet focused on identifying microbial agents driving methane oxidation. Although methanotrophs were rare, we identified three uncultivated groups affiliated with particulate methane monooxygenase (pMMO encoding phylogenetic groups (OPU, and methanotrophic symbionts as primary drivers of methane oxidation in Saanich Inlet. Distribution and activity patterns for these three groups were consistent with niche partitioning that became increasingly resolved during water column stratification. Moreover co-occurrence analysis combined with multi-level indicator species analysis revealed significant correlations between operational taxonomic units affiliated with Methylophaga, Methylophilales, SAR324, Verrucomicrobia, and Planctomycetes with OPUs and methanotrophic symbiont groups. Taken together these observations shed new light on the composition, dynamics, and potential interspecific interactions of microbes associated with CH4 cycling in the Saanich Inlet water column, provide a baseline for comparison between coastal and open ocean OMZs and support the potential role of OPUs, and methanotrophic symbiont groups as a widely distributed pelagic sink for CH4 along continental margins.

  20. Effect of Mineral Nutrients on the Kinetics of Methane Utilization by Methanotrophs

    DEFF Research Database (Denmark)

    Boiesen, Anette; Arvin, Erik; Broholm, Kim

    1993-01-01

    The effect of different mineral nutrients on the kinetics of methane biodegradation by a mixed culture of methanotrophic bacteria was studied. The substrate factors examined were ammonia, iron, copper, manganese, phosphate, and sulphide. The presence of iron in the growth medium had a strong effect...

  1. Ammonium addition inhibits 13C-methane incorporation into methanotroph membrane lipids in a freshwater sediment

    NARCIS (Netherlands)

    Nold, s.c.; Boschker, H.T.S.; Pel, R.; Laanbroek, H.J.

    1999-01-01

    To investigate the effect of ammonium addition on the species composition and activity of freshwater methane oxidizing bacteria, intact sediment cores were labeled with (CH4)-C-13 and incubated under ambient and elevated ammonium concentrations. After 7 days, methanotroph activity was assessed by qu

  2. Enhanced bioleaching on attachment of indigenous acidophilic bacteria to pyrite surface

    Science.gov (United States)

    Wi, D. W.; Cho, K. H.; Kim, B. J.; Choi, N. C.; Park, C. Y.

    2012-04-01

    In recent years, bioleaching has been widely applied on an industrial scale due to the advantages of low cost and environment friendliness. The direct contact mechanism of bioleaching assumes the action of a metal sulfide-attached cell oxidizing the mineral by an enzyme system with oxygen to sulfate and metal cations. Fundamental surface properties of sulfide particles and leaching-bacteria in bioleaching play the key role in the efficiency of this process. The aim of this work is to investigate of direct contact bioleaching mechanism on pyrite through attachment properties between indigenous acidophilic bacteria and pyrite surfaces. The bacteria were obtained from sulfur hot springs, Hatchobaru thermal electricity plant in Japan. And pyrite was collected from mine waste from Gwang-yang abandoned gold mines, Korea. In XRD analyses of the pyrite, x-ray diffracted d-value belong to pyrite was observed. The indigenous acidophilic bacteria grew well in a solution and over the course of incubation pH decreased and Eh increased. In relation to a bacterial growth-curve, the lag phase was hardly shown while the exponential phase was very fast. Bioleaching experiment result was showed that twenty days after the indigenous acidophilic bacteria were inoculated to a pyrite-leaching medium, the bacterial sample had a greater concentration of Fe and Zn than within the control sample. In SEM-EDS analyses, rod-shaped bacteria and round-shaped microbes were well attached to the surface of pyrite. The size of the rod-shaped bacteria ranged from 1.05~1.10 ? to 4.01~5.38 ?. Round-shaped microbes were more than 3.0 ? in diameter. Paired cells of rod-shaped bacteria were attached to the surface of pyrite linearly.

  3. Transfer of IncP Plasmids to Extremely Acidophilic Thiobacillus thiooxidans

    OpenAIRE

    Jin, S. M.; Yan, W. M.; Wang, Z. N.

    1992-01-01

    The broad-host-range IncP plasmids RP4, R68.45, RP1::Tn501, and and pUB307 were transferred directly to extremely acidophilic Thiobacillus thiooxidans from Escherichia coli by conjugation at frequencies of 10-5 to 10-7 per recipient. The ability of T. thiooxidans to receive and express the antibiotic resistance markers was examined. The plasmid RP4 was transferred back to E. coli from T. thiooxidans at a frequency of 1.0 × 10-3 per recipient.

  4. Transfer of IncP Plasmids to Extremely Acidophilic Thiobacillus thiooxidans.

    Science.gov (United States)

    Jin, S M; Yan, W M; Wang, Z N

    1992-01-01

    The broad-host-range IncP plasmids RP4, R68.45, RP1::Tn501, and and pUB307 were transferred directly to extremely acidophilic Thiobacillus thiooxidans from Escherichia coli by conjugation at frequencies of 10 to 10 per recipient. The ability of T. thiooxidans to receive and express the antibiotic resistance markers was examined. The plasmid RP4 was transferred back to E. coli from T. thiooxidans at a frequency of 1.0 x 10 per recipient.

  5. [Preparation of Copper and Nickel from Metallurgical Waste Products with the Use of Acidophilic Chemolithotrophic Microorganisms].

    Science.gov (United States)

    Fomchenko, N V; Murav'ev, M I

    2015-01-01

    The study concerns the leaching of copper, nickel, and cobalt from metallurgical production slag with trivalent iron sulphates prepared in the process of oxidation of bivalent iron ions with the use of associations of acidophilic chemolithotrophic microorganisms. At the same time, copper extraction in the solution reached 91.2%, nickel reached 74.9%, and cobalt reached 90.1%. Copper was extracted by cementation, and nickel as sulphate was extracted by electrolysis. Associations of microorganisms can then completely bioregenerate the solution obtained after leaching.

  6. Cytochrome 572 is a conspicuous membrane protein with iron oxidation activity purified directly from a natural acidophilic microbial community.

    Science.gov (United States)

    Jeans, Chris; Singer, Steven W; Chan, Clara S; Verberkmoes, Nathan C; Shah, Manesh; Hettich, Robert L; Banfield, Jillian F; Thelen, Michael P

    2008-05-01

    Recently, there has been intense interest in the role of electron transfer by microbial communities in biogeochemical systems. We examined the process of iron oxidation by microbial biofilms in one of the most extreme environments on earth, where the inhabited water is pH 0.5-1.2 and laden with toxic metals. To approach the mechanism of Fe(II) oxidation as a means of cellular energy acquisition, we isolated proteins from natural samples and found a conspicuous and novel cytochrome, Cyt(572), which is unlike any known cytochrome. Both the character of its covalently bound prosthetic heme group and protein sequence are unusual. Extraction of proteins directly from environmental biofilm samples followed by membrane fractionation, detergent solubilization and gel filtration chromatography resulted in the purification of an abundant yellow-red protein. The purified protein has a cytochrome c-type heme binding motif, CxxCH, but a unique spectral signature at 572 nm, and thus is called Cyt(572). It readily oxidizes Fe(2+) in the physiologically relevant acidic regime, from pH 0.95-3.4. Other physical characteristics are indicative of a membrane-bound multimeric protein. Circular dichroism spectroscopy indicates that the protein is largely beta-stranded, and 2D Blue-Native polyacrylamide gel electrophoresis and chemical crosslinking independently point to a multi-subunit structure for Cyt(572). By analyzing environmental genomic information from biofilms in several distinctly different mine locations, we found multiple genetic variants of Cyt(572). MS proteomics of extracts from these biofilms substantiated the prevalence of these variants in the ecosystem. Due to its abundance, cellular location and Fe(2+) oxidation activity at very low pH, we propose that Cyt(572) provides a critical function for fitness within the ecological niche of these acidophilic microbial communities.

  7. An alternative allosteric regulation mechanism of an acidophilic l-lactate dehydrogenase from Enterococcus mundtii 15-1A

    Directory of Open Access Journals (Sweden)

    Yasuyuki Matoba

    2014-01-01

    Full Text Available A plant-derived Enterococcus mundtii 15-1A, that has been previously isolated from Brassica rapa L. subsp. nipposinica (L.H. Bailey Hanelt var. linearifolia by our group, possesses two kinds of l-lactate dehydrogenase (l-LDH: LDH-1 and LDH-2. LDH-1 was activated under low concentration of fluctose-1,6-bisphosphate (FBP at both pH 5.5 and 7.5. Although LDH-2 was also activated under the low concentration of FBP at pH 5.5, a high concentration of FBP is necessary to activate it at pH 7.5. The present study shows the crystal structures of the acidophilic LDH-2 in a complex with and without FBP and NADH. Although the tertiary structure of the ligands-bound LDH-2 is similar to that of the active form of other bacterial l-LDHs, the structure without the ligands is different from that of any other previously determined l-LDHs. Major structural alterations between the two structures of LDH-2 were observed at two regions in one subunit. At the N-terminal parts of the two regions, the ligands-bound form takes an α-helical structure, while the form without ligands displays more disordered and extended structures. A vacuum-ultraviolet circular dichroism analysis showed that the α-helix content of LDH-2 in solution is approximately 30% at pH 7.5, which is close to that in the crystal structure of the form without ligands. A D241N mutant of LDH-2, which was created by us to easily form an α-helix at one of the two parts, exhibited catalytic activity even in the absence of FBP at both pH 5.5 and 7.5.

  8. 极端环境下嗜热酸甲烷营养细菌研究进展%Advances in thermoacidophilic methanotrophs from extreme environments

    Institute of Scientific and Technical Information of China (English)

    郑勇; 郑袁明; 张丽梅; 贺纪正

    2009-01-01

    Methane-oxidizing bacteria (methanotrophs) play an important role in the biogeochemical carbon cycle and in controlling global climate change, by converting methane to carbon dioxide or biomass. Although these bacteria have been isolated from a variety of environments, most of which grow best at neutral pH (5-8) and moderate temperature ranges (20-35℃). Based on the phylogenetic analysis, methanotrophs are classified into type I and type II, which belong to the gamma- and alpha-Proteobacteria, respectively. Very recently, three independent studies have isolated methane-oxidizing microorganisms from extreme thermoacidophilic environments with pH values of approximately 1 and temperatures higher than 50℃, these nonproteobacterial strains were all identified as members of the phylum Verrucomicrobia. These new and unusual studies will undoubtedly expand the known phylogenetic and functional diversity of methanotrophs, also indicate that novel methane oxidizing pathways and mechanisms could exist in the methanotrophs. This review illustrates the latest advances in thermoacidophilic methanotrophs, based on the recent three reports on methane oxidation in the extreme environments.%甲烷营养细菌能够将温室气体甲烷(CH4)转化为CO2或生物质,在碳生物地球化学循环及缓解由温室气体导致的全球气候变化方面发挥着重要的作用.甲烷营养细菌生存的条件范围较为广泛,但在中性pH (5~8)和中温(20~35℃)范围内生长最佳.系统进化分析认为,它们均属于γ-或α-变形菌门(Proteobacteria).最近3项独立完成的研究从极端热酸(pH接近1,温度高于50℃)环境中分离获得了具有甲烷氧化(营养)功能的微生物,经鉴定均属于疣微菌门(Verrucomicrobia).这些全新的、不同于以往的研究结果不仅是对现有关于甲烷营养细菌生态学认知的进一步拓展,同时也暗示着可能存在着新型的、由微生物介导的CH4氧化途径与机制. 因此,特

  9. Low energy emulsion-based fermentation enabling accelerated methane mass transfer and growth of poly(3-hydroxybutyrate)-accumulating methanotrophs.

    Science.gov (United States)

    Myung, Jaewook; Kim, Minkyu; Pan, Ming; Criddle, Craig S; Tang, Sindy K Y

    2016-05-01

    Methane is a low-cost feedstock for the production of polyhydroxyalkanoate biopolymers, but methanotroph fermentations are limited by the low solubility of methane in water. To enhance mass transfer of methane to water, vigorous mixing or agitation is typically used, which inevitably increases power demand and operational costs. This work presents a method for accelerating methane mass transfer without agitation by growing methanotrophs in water-in-oil emulsions, where the oil has a higher solubility for methane than water does. In systems without agitation, the growth rate of methanotrophs in emulsions is five to six times that of methanotrophs in the medium-alone incubations. Within seven days, cells within the emulsions accumulate up to 67 times more P3HB than cells in the medium-alone incubations. This is achieved due to the increased interfacial area of the aqueous phase, and accelerated methane diffusion through the oil phase.

  10. Auto- and heterotrophic acidophilic bacteria enhance the bioremediation efficiency of sediments contaminated by heavy metals.

    Science.gov (United States)

    Beolchini, Francesca; Dell'Anno, Antonio; De Propris, Luciano; Ubaldini, Stefano; Cerrone, Federico; Danovaro, Roberto

    2009-03-01

    This study deals with bioremediation treatments of dredged sediments contaminated by heavy metals based on the bioaugmentation of different bacterial strains. The efficiency of the following bacterial consortia was compared: (i) acidophilic chemoautotrophic, Fe/S-oxidising bacteria, (ii) acidophilic heterotrophic bacteria able to reduce Fe/Mn fraction, co-respiring oxygen and ferric iron and (iii) the chemoautotrophic and heterotrophic bacteria reported above, pooled together, as it was hypothesised that the two strains could cooperate through a mutual substrate supply. The effect of the bioremediation treatment based on the bioaugmentation of Fe/S-oxidising strains alone was similar to the one based only on Fe-reducing bacteria, and resulted in heavy-metal extraction yields typically ranging from 40% to 50%. The efficiency of the process based only upon autotrophic bacteria was limited by sulphur availability. However, when the treatment was based on the addition of Fe-reducing bacteria and the Fe/S oxidizing bacteria together, their growth rates and efficiency in mobilising heavy metals increased significantly, reaching extraction yields >90% for Cu, Cd, Hg and Zn. The additional advantage of the new bioaugmentation approach proposed here is that it is independent from the availability of sulphur. These results open new perspectives for the bioremediation technology for the removal of heavy metals from highly contaminated sediments.

  11. Electricity generation from an inorganic sulfur compound containing mining wastewater by acidophilic microorganisms.

    Science.gov (United States)

    Ni, Gaofeng; Christel, Stephan; Roman, Pawel; Wong, Zhen Lim; Bijmans, Martijn F M; Dopson, Mark

    2016-09-01

    Sulfide mineral processing often produces large quantities of wastewaters containing acid-generating inorganic sulfur compounds. If released untreated, these wastewaters can cause catastrophic environmental damage. In this study, microbial fuel cells were inoculated with acidophilic microorganisms to investigate whether inorganic sulfur compound oxidation can generate an electrical current. Cyclic voltammetry suggested that acidophilic microorganisms mediated electron transfer to the anode, and that electricity generation was catalyzed by microorganisms. A cation exchange membrane microbial fuel cell, fed with artificial wastewater containing tetrathionate as electron donor, reached a maximum whole cell voltage of 72 ± 9 mV. Stepwise replacement of the artificial anolyte with real mining process wastewater had no adverse effect on bioelectrochemical performance and generated a maximum voltage of 105 ± 42 mV. 16S rRNA gene sequencing of the microbial consortia resulted in sequences that aligned within the genera Thermoplasma, Ferroplasma, Leptospirillum, Sulfobacillus and Acidithiobacillus. This study opens up possibilities to bioremediate mining wastewater using microbial fuel cell technology.

  12. Selection of Type I and Type II Methanotrophic Proteobacteria in a Fluidized Bed Reactor under Non-Sterile Conditions

    Science.gov (United States)

    2011-08-01

    00-00-2011 to 00-00-2011 4. TITLE AND SUBTITLE Selection of Type I and Type II methanotrophic proteobacteria in a fluidized bed reactor under...laboratory- scale fluidized bed reactor was initially inoculated with a Type II Methylocystis-like dominated culture. At elevated levels of dissolved...personal copy Selection of Type I and Type II methanotrophic proteobacteria in a fluidized bed reactor under non-sterile conditions Andrew R. Pfluger a, Wei

  13. Salinity Affects the Composition of the Aerobic Methanotroph Community in Alkaline Lake Sediments from the Tibetan Plateau.

    Science.gov (United States)

    Deng, Yongcui; Liu, Yongqin; Dumont, Marc; Conrad, Ralf

    2017-01-01

    Lakes are widely distributed on the Tibetan Plateau, which plays an important role in natural methane emission. Aerobic methanotrophs in lake sediments reduce the amount of methane released into the atmosphere. However, no study to date has analyzed the methanotroph community composition and their driving factors in sediments of these high-altitude lakes (>4000 m). To provide new insights on this aspect, the abundance and composition in the sediments of six high-altitude alkaline lakes (including both freshwater and saline lakes) on the Tibetan Plateau were studied. The quantitative PCR, terminal restriction fragment length polymorphism, and 454-pyrosequencing methods were used to target the pmoA genes. The pmoA gene copies ranged 10(4)-10(6) per gram fresh sediment. Type I methanotrophs predominated in Tibetan lake sediments, with Methylobacter and uncultivated type Ib methanotrophs being dominant in freshwater lakes and Methylomicrobium in saline lakes. Combining the pmoA-pyrosequencing data from Tibetan lakes with other published pmoA-sequencing data from lake sediments of other regions, a significant salinity and alkalinity effect (P = 0.001) was detected, especially salinity, which explained ∼25% of methanotroph community variability. The main effect was Methylomicrobium being dominant (up to 100%) in saline lakes only. In freshwater lakes, however, methanotroph composition was relatively diverse, including Methylobacter, Methylocystis, and uncultured type Ib clusters. This study provides the first methanotroph data for high-altitude lake sediments (>4000 m) and shows that salinity is a driving factor for the community composition of aerobic methanotrophs.

  14. A metagenomic study of methanotrophic microorganisms in Coal Oil Point seep sediments

    Directory of Open Access Journals (Sweden)

    Haverkamp Thomas HA

    2011-10-01

    Full Text Available Abstract Background Methane oxidizing prokaryotes in marine sediments are believed to function as a methane filter reducing the oceanic contribution to the global methane emission. In the anoxic parts of the sediments, oxidation of methane is accomplished by anaerobic methanotrophic archaea (ANME living in syntrophy with sulphate reducing bacteria. This anaerobic oxidation of methane is assumed to be a coupling of reversed methanogenesis and dissimilatory sulphate reduction. Where oxygen is available aerobic methanotrophs take part in methane oxidation. In this study, we used metagenomics to characterize the taxonomic and metabolic potential for methane oxidation at the Tonya seep in the Coal Oil Point area, California. Two metagenomes from different sediment depth horizons (0-4 cm and 10-15 cm below sea floor were sequenced by 454 technology. The metagenomes were analysed to characterize the distribution of aerobic and anaerobic methanotrophic taxa at the two sediment depths. To gain insight into the metabolic potential the metagenomes were searched for marker genes associated with methane oxidation. Results Blast searches followed by taxonomic binning in MEGAN revealed aerobic methanotrophs of the genus Methylococcus to be overrepresented in the 0-4 cm metagenome compared to the 10-15 cm metagenome. In the 10-15 cm metagenome, ANME of the ANME-1 clade, were identified as the most abundant methanotrophic taxon with 8.6% of the reads. Searches for particulate methane monooxygenase (pmoA and methyl-coenzyme M reductase (mcrA, marker genes for aerobic and anaerobic oxidation of methane respectively, identified pmoA in the 0-4 cm metagenome as Methylococcaceae related. The mcrA reads from the 10-15 cm horizon were all classified as originating from the ANME-1 clade. Conclusions Most of the taxa detected were present in both metagenomes and differences in community structure and corresponding metabolic potential between the two samples were mainly

  15. Methanotrophic communities in aerobic soils with different stages of natural reforestation

    Science.gov (United States)

    Kravchenko, Irina; Sukhacheva, Marina; Kuznetsova, Tatyana

    2017-04-01

    The land use and management has a significant impact on global biogeochemical cycles of biogenic elements and the Earth's climate. Deforestation is known to change the soil from a net sink for CH4 to a net source as a result of alteration in the activity and composition of the methanotrophic communities. At the same time, the recovery of ecosystems after their withdrawal from agricultural use is poorly understood. Currently, in Russia, the former arable lands occupy about 20% of the territory and more than half of them are not used in agriculture since the early 90-s. Here, soil CH4 oxidation rates and associated methanotrophic communities were examined in a chronosequence of gray forest soils, Moscow region, Russia, consisting of cropland soils, soils at the different stages under postagrogenic forest regenerating, and in a mature native forest. CH4 concentrations were measured by GC and several chemical (pH, total C and N, NH4 -N and NO3 -N) and physical (moisture content, porosity, water-filled pore space and bulk density) soil properties were evaluated. Methane oxidation rates were significantly influenced by reforestation and the regenerating soils have the potential to reach those of the native forest. In fallow, shrublands and young forest soil CH4-oxidation rates were significantly higher as compared with cropland, but not fully stabilized even after 25 years of reforestation. To examine whether changes in CH4-oxidation rate were linked to a shift in the microbial community, we have analyzed soil methanotrophic communities by cloning and sequencing of particulate methane monooxygenase (pmoA) using the primer pair A189-mb650. Based on the relative proportion of the clones it was shown the dominance Type II related and uncultured methanotrophs in forest soils. Both Type I and Type II methanotrophs were found in arable and postagrogenic soils, and the relative abundance of Type II methanotrophs increased with the age of regeneration and recovered after 15

  16. Hydrology is reflected in the functioning and community composition of methanotrophs in the littoral wetland of a boreal lake.

    Science.gov (United States)

    Siljanen, Henri M P; Saari, Anne; Krause, Sascha; Lensu, Anssi; Abell, Guy C J; Bodrossy, Levente; Bodelier, Paul L E; Martikainen, Pertti J

    2011-03-01

    In lake ecosystems a major proportion of methane (CH(4) ) emissions originate from the littoral zone, which can have a great spatial variability in hydrology, soil quality and vegetation. Hitherto, spatial heterogeneity and the effects it has on functioning and diversity of methanotrophs in littoral wetlands have been poorly understood. A diagnostic microarray based on the particulate methane monooxygenase gene coupled with geostatistics was used to analyse spatial patterns of methanotrophs in the littoral wetland of a eutrophic boreal lake (Lake Kevätön, Eastern Finland). The wetland had a hydrology gradient with a mean water table varying from -8 to -25 cm. The wettest area, comprising the highest CH(4) oxidation, had the highest abundance and species richness of methanotrophs. A high water table favoured the occurrence of type Ib methanotrophs, whereas types Ia and II were found under all moisture conditions. Thus the spatial heterogeneity in functioning and diversity of methanotrophs in littoral wetlands is highly dependent on the water table, which in turn varies spatially in relation to the geomorphology of the wetland. We suggest that changes in water levels resulting from regulation of lakes and/or global change will affect the abundance, activity and diversity of methanotrophs, and consequently CH(4) emissions from such systems. © 2010 Federation of European Microbiological Societies. Published by Blackwell Publishing Ltd. All rights reserved.

  17. [Continuous biosynthesis of epoxypropane in a methanotrophic attached-films reactor].

    Science.gov (United States)

    Xin, Jia-Ying; Cui, Jun-Ru; Chen, Jian-Bo; Li, Shu-Ben; Xia, Chun-Gu

    2002-01-01

    Using a fluidized bed as immobilization system, mixed culture methanotrophic attached-films were developed on diatomite particles. The Methane Monooxygenase (MMO) activity was found to increase obviously as soon as the lag phase ended. Greater than 90% of the MMO activity in the bed was attached. Biofilm concentration of 3.3-3.7 mg dry weight cell/g DS was observed. Batch experiments were performed to explore the possibility of producing epoxypropane by a cooxidation process. The effect of methane on the oxidation of propene to epoxypropane and the effect of propene on the growth of methanotroph were also studied. In continuous experiments, optimum mixed gaseous substrates (methane: 35%; propene: 20%; oxygen: 45%) were continuously circulated through the fluidized bed reactor to remove product. Initial epoxypropane productivity was 110-150 mumol/d. The bioreactor operated continuously for 25 d without obvious loss of epoxypropane productivity.

  18. Potential of pmoA amplicon pyrosequencing for methanotroph diversity studies.

    Science.gov (United States)

    Lüke, Claudia; Frenzel, Peter

    2011-09-01

    We analyzed the potential of pmoA amplicon pyrosequencing compared to that of Sanger sequencing with paddy soils as a model environment. We defined operational taxonomic unit (OTU) cutoff values of 7% and 18%, reflecting methanotrophic species and major phylogenetic pmoA lineages, respectively. Major lineages were already well covered by clone libraries; nevertheless, pyrosequencing provided a higher level of diversity at the species level.

  19. Molecular Analysis of the pmo (Particulate Methane Monooxygenase) Operons from Two Type II Methanotrophs

    OpenAIRE

    Gilbert, Bettina; McDonald, Ian R.; Finch, Ruth; Stafford, Graham P.; Nielsen, Allan K.; Murrell, J. Colin

    2000-01-01

    The particulate methane monooxygenase gene clusters, pmoCAB, from two representative type II methanotrophs of the α-Proteobacteria, Methylosinus trichosporium OB3b and Methylocystis sp. strain M, have been cloned and sequenced. Primer extension experiments revealed that the pmo cluster is probably transcribed from a single transcriptional start site located 300 bp upstream of the start of the first gene, pmoC, for Methylocystis sp. strain M. Immediately upstream of the putative start site, co...

  20. Molecular analysis of the pmo (particulate methane monooxygenase) operons from two type II methanotrophs.

    Science.gov (United States)

    Gilbert, B; McDonald, I R; Finch, R; Stafford, G P; Nielsen, A K; Murrell, J C

    2000-03-01

    The particulate methane monooxygenase gene clusters, pmoCAB, from two representative type II methanotrophs of the alpha-Proteobacteria, Methylosinus trichosporium OB3b and Methylocystis sp. strain M, have been cloned and sequenced. Primer extension experiments revealed that the pmo cluster is probably transcribed from a single transcriptional start site located 300 bp upstream of the start of the first gene, pmoC, for Methylocystis sp. strain M. Immediately upstream of the putative start site, consensus sequences for sigma(70) promoters were identified, suggesting that these pmo genes are recognized by sigma(70) and negatively regulated under low-copper conditions. The pmo genes were cloned in several overlapping fragments, since parts of these genes appeared to be toxic to the Escherichia coli host. Methanotrophs contain two virtually identical copies of pmo genes, and it was necessary to use Southern blotting and probing with pmo gene fragments in order to differentiate between the two pmoCAB clusters in both methanotrophs. The complete DNA sequence of one copy of pmo genes from each organism is reported here. The gene sequences are 84% similar to each other and 75% similar to that of a type I methanotroph of the gamma-Proteobacteria, Methylococcus capsulatus Bath. The derived proteins PmoC and PmoA are predicted to be highly hydrophobic and consist mainly of transmembrane-spanning regions, whereas PmoB has only two putative transmembrane-spanning helices. Hybridization experiments showed that there are two copies of pmoC in both M. trichosporium OB3b and Methylocystis sp. strain M, and not three copies as found in M. capsulatus Bath.

  1. Conventional methanotrophs are responsible for atmospheric methane oxidation in paddy soils

    Science.gov (United States)

    Cai, Yuanfeng; Zheng, Yan; Bodelier, Paul L. E.; Conrad, Ralf; Jia, Zhongjun

    2016-06-01

    Soils serve as the biological sink of the potent greenhouse gas methane with exceptionally low concentrations of ~1.84 p.p.m.v. in the atmosphere. The as-yet-uncultivated methane-consuming bacteria have long been proposed to be responsible for this `high-affinity' methane oxidation (HAMO). Here we show an emerging HAMO activity arising from conventional methanotrophs in paddy soil. HAMO activity was quickly induced during the low-affinity oxidation of high-concentration methane. Activity was lost gradually over 2 weeks, but could be repeatedly regained by flush-feeding the soil with elevated methane. The induction of HAMO activity occurred only after the rapid growth of methanotrophic populations, and a metatranscriptome-wide association study suggests that the concurrent high- and low-affinity methane oxidation was catalysed by known methanotrophs rather than by the proposed novel atmospheric methane oxidizers. These results provide evidence of atmospheric methane uptake in periodically drained ecosystems that are typically considered to be a source of atmospheric methane.

  2. Starvation and recovery in the deep-sea methanotroph Methyloprofundus sedimenti.

    Science.gov (United States)

    Tavormina, Patricia L; Kellermann, Matthias Y; Antony, Chakkiath Paul; Tocheva, Elitza I; Dalleska, Nathan F; Jensen, Ashley J; Valentine, David L; Hinrichs, Kai-Uwe; Jensen, Grant J; Dubilier, Nicole; Orphan, Victoria J

    2017-01-01

    In the deep ocean, the conversion of methane into derived carbon and energy drives the establishment of diverse faunal communities. Yet specific biological mechanisms underlying the introduction of methane-derived carbon into the food web remain poorly described, due to a lack of cultured representative deep-sea methanotrophic prokaryotes. Here, the response of the deep-sea aerobic methanotroph Methyloprofundus sedimenti to methane starvation and recovery was characterized. By combining lipid analysis, RNA analysis, and electron cryotomography, it was shown that M. sedimenti undergoes discrete cellular shifts in response to methane starvation, including changes in headgroup-specific fatty acid saturation levels, and reductions in cytoplasmic storage granules. Methane starvation is associated with a significant increase in the abundance of gene transcripts pertinent to methane oxidation. Methane reintroduction to starved cells stimulates a rapid, transient extracellular accumulation of methanol, revealing a way in which methane-derived carbon may be routed to community members. This study provides new understanding of methanotrophic responses to methane starvation and recovery, and lays the initial groundwork to develop Methyloprofundus as a model chemosynthesizing bacterium from the deep sea. © 2016 John Wiley & Sons Ltd.

  3. Response of methanotrophic communities to afforestation and reforestation in New Zealand.

    Science.gov (United States)

    Nazaries, Loïc; Tate, Kevin R; Ross, Des J; Singh, Jagrati; Dando, John; Saggar, Surinder; Baggs, Elizabeth M; Millard, Peter; Murrell, J Colin; Singh, Brajesh K

    2011-11-01

    Methanotrophs use methane (CH(4)) as a carbon source. They are particularly active in temperate forest soils. However, the rate of change of CH(4) oxidation in soil with afforestation or reforestation is poorly understood. Here, soil CH(4) oxidation was examined in New Zealand volcanic soils under regenerating native forests following burning, and in a mature native forest. Results were compared with data for pasture to pine land-use change at nearby sites. We show that following soil disturbance, as little as 47 years may be needed for development of a stable methanotrophic community similar to that in the undisturbed native forest soil. Corresponding soil CH(4)-oxidation rates in the regenerating forest soil have the potential to reach those of the mature forest, but climo-edaphic fators appear limiting. The observed changes in CH(4)-oxidation rate were directly linked to a prior shift in methanotrophic communities, which suggests microbial control of the terrestrial CH(4) flux and identifies the need to account for this response to afforestation and reforestation in global prediction of CH(4) emission.

  4. Monitoring Acidophilic Microbes with Real-Time Polymerase Chain Reaction (PCR) Assays

    Energy Technology Data Exchange (ETDEWEB)

    Frank F. Roberto

    2008-08-01

    Many techniques that are used to characterize and monitor microbial populations associated with sulfide mineral bioleaching require the cultivation of the organisms on solid or liquid media. Chemolithotrophic species, such as Acidithiobacillus ferrooxidans and Leptospirillum ferrooxidans, or thermophilic chemolithotrophs, such as Acidianus brierleyi and Sulfolobus solfataricus can grow quite slowly, requiring weeks to complete efforts to identify and quantify these microbes associated with bioleach samples. Real-time PCR (polymerase chain reaction) assays in which DNA targets are amplified in the presence of fluorescent oligonucleotide primers, allowing the monitoring and quantification of the amplification reactions as they progress, provide a means of rapidly detecting the presence of microbial species of interest, and their relative abundance in a sample. This presentation will describe the design and use of such assays to monitor acidophilic microbes in the environment and in bioleaching operations. These assays provide results within 2-3 hours, and can detect less than 100 individual microbial cells.

  5. Optimized Production of Xylitol from Xylose Using a Hyper-Acidophilic Candida tropicalis

    Directory of Open Access Journals (Sweden)

    Elena Tamburini

    2015-08-01

    Full Text Available The yeast Candida tropicalis DSM 7524 produces xylitol, a natural, low-calorie sweetener, by fermentation of xylose. In order to increase xylitol production rate during the submerged fermentation process, some parameters-substrate (xylose concentration, pH, aeration rate, temperature and fermentation strategy-have been optimized. The maximum xylitol yield reached at 60–80 g/L initial xylose concentration, pH 5.5 at 37 °C was 83.66% (w/w on consumed xylose in microaerophilic conditions (kLa = 2·h−1. Scaling up on 3 L fermenter, with a fed-batch strategy, the best xylitol yield was 86.84% (w/w, against a 90% of theoretical yield. The hyper-acidophilic behaviour of C. tropicalis makes this strain particularly promising for industrial application, due to the possibility to work in non-sterile conditions.

  6. In-Situ Quantification of Methanotrophic Activity in a Landfill Cover Soil Using Gas Push-Pull Tests

    Science.gov (United States)

    Gomez, K. E.; Gonzalez-Gil, G.; Schroth, M. H.; Zeyer, J.

    2007-12-01

    Landfills are both a major anthropogenic source and a sink for the greenhouse gas CH4. Methanogenic bacteria produce CH4 during the anaerobic digestion of landfill waste, whereas, methanotrophic bacteria consume CH4 as it is transported through a landfill cover soil. Methanotrophs are thought to be ubiquitous in soils, but typically exist in large numbers at oxic/anoxic interfaces, close to anaerobic methane sources but exposed to oxygen required for metabolism. Accurate in-situ quantification of the sink strength of methanotrophs in landfill cover soils is needed for global carbon balances and for local emissions mitigation strategies. We measured in-situ CH4 concentrations at 30, 60, and 100 cm depth at 18 evenly spaced locations across a landfill cover soil. Furthermore, we performed Gas Push-Pull Tests (GPPTs) to estimate in-situ rates of methanotrophic activity in the cover soil. The GPPT is a gas-tracer test in which a gas mixture containing CH4, O2, and non-reactive tracer gases is injected (pushed) into the soil followed by extraction (pull) from the same location. Quantification of CH4 oxidation rates is based upon comparison of the breakthrough curves of CH4 and tracer gases. We present the results of a series of GPPTs conducted at two locations in the cover soil to assess the feasibility and reproducibility of this technique to quantify methanotrophic activity. Additional GPPTs were performed with a methanotrophic inhibitor in the injection gas mixture to confirm the appropriate choice of tracers to quantify CH4 oxidation. Estimated CH4 oxidation rate constants indicate that the cover soil contains a highly active methanotrophic community.

  7. Effect of trichloroethylene and tetrachloroethylene on methane oxidation and community structure of methanotrophic consortium.

    Science.gov (United States)

    Choi, Sun-Ah; Lee, Eun-Hee; Cho, Kyung-Suk

    2013-01-01

    The methane oxidation rate and community structure of a methanotrophic consortium were analyzed to determine the effects of trichloroethylene (TCE) and tetrachloroethylene (PCE) on methane oxidation. The maximum methane oxidation rate (Vmax ) of the consortium was 326.8 μmol·g-dry biomass(-1)·h(-1), and it had a half-saturation constant (Km ) of 143.8 μM. The addition of TCE or PCE resulted in decreased methane oxidation rates, which were decreased from 101.73 to 5.47-24.64 μmol·g-dry biomass(-1)·h(-1) with an increase in the TCE-to-methane ratio, and to 61.95-67.43 μmol·g-dry biomass(-1)·h(-1) with an increase in the PCE-to-methane ratio. TCE and PCE were non-competitive inhibitors for methane oxidation, and their inhibition constants (Ki ) were 33.4 and 132.0 μM, respectively. When the methanotrophic community was analyzed based on pmoA using quantitative real-time PCR (qRT-PCR), the pmoA gene copy numbers were shown to decrease from 7.3 ± 0.7 × 10(8) to 2.1-5.0 × 10(7) pmoA gene copy number · g-dry biomass(-1) with an increase in the TCE-to-methane ratio and to 2.5-7.0 × 10(7) pmoA gene copy number · g-dry biomass(-1) with an increase in the PCE-to-methane ratio. Community analysis by microarray demonstrated that Methylocystis (type II methanotrophs) were the most abundant in the methanotrophic community composition in the presence of TCE. These results suggest that toxic effects caused by TCE and PCE change not only methane oxidation rates but also the community structure of the methanotrophic consortium.

  8. Quantitative proteomic analyses of the response of acidophilic microbial communities to different pH conditions.

    Science.gov (United States)

    Belnap, Christopher P; Pan, Chongle; Denef, Vincent J; Samatova, Nagiza F; Hettich, Robert L; Banfield, Jillian F

    2011-07-01

    Extensive genomic characterization of multi-species acid mine drainage microbial consortia combined with laboratory cultivation has enabled the application of quantitative proteomic analyses at the community level. In this study, quantitative proteomic comparisons were used to functionally characterize laboratory-cultivated acidophilic communities sustained in pH 1.45 or 0.85 conditions. The distributions of all proteins identified for individual organisms indicated biases for either high or low pH, and suggests pH-specific niche partitioning for low abundance bacteria and archaea. Although the proteome of the dominant bacterium, Leptospirillum group II, was largely unaffected by pH treatments, analysis of functional categories indicated proteins involved in amino acid and nucleotide metabolism, as well as cell membrane/envelope biogenesis were overrepresented at high pH. Comparison of specific protein abundances indicates higher pH conditions favor Leptospirillum group III, whereas low pH conditions promote the growth of certain archaea. Thus, quantitative proteomic comparisons revealed distinct differences in community composition and metabolic function of individual organisms during different pH treatments. Proteomic analysis revealed other aspects of community function. Different numbers of phage proteins were identified across biological replicates, indicating stochastic spatial heterogeneity of phage outbreaks. Additionally, proteomic data were used to identify a previously unknown genotypic variant of Leptospirillum group II, an indication of selection for a specific Leptospirillum group II population in laboratory communities. Our results confirm the importance of pH and related geochemical factors in fine-tuning acidophilic microbial community structure and function at the species and strain level, and demonstrate the broad utility of proteomics in laboratory community studies.

  9. Architecture and gene repertoire of the flexible genome of the extreme acidophile Acidithiobacillus caldus.

    Directory of Open Access Journals (Sweden)

    Lillian G Acuña

    Full Text Available BACKGROUND: Acidithiobacillus caldus is a sulfur oxidizing extreme acidophile and the only known mesothermophile within the Acidithiobacillales. As such, it is one of the preferred microbes for mineral bioprocessing at moderately high temperatures. In this study, we explore the genomic diversity of A. caldus strains using a combination of bioinformatic and experimental techniques, thus contributing first insights into the elucidation of the species pangenome. PRINCIPAL FINDINGS: Comparative sequence analysis of A. caldus ATCC 51756 and SM-1 indicate that, despite sharing a conserved and highly syntenic genomic core, both strains have unique gene complements encompassing nearly 20% of their respective genomes. The differential gene complement of each strain is distributed between the chromosomal compartment, one megaplasmid and a variable number of smaller plasmids, and is directly associated to a diverse pool of mobile genetic elements (MGE. These include integrative conjugative and mobilizable elements, genomic islands and insertion sequences. Some of the accessory functions associated to these MGEs have been linked previously to the flexible gene pool in microorganisms inhabiting completely different econiches. Yet, others had not been unambiguously mapped to the flexible gene pool prior to this report and clearly reflect strain-specific adaption to local environmental conditions. SIGNIFICANCE: For many years, and because of DNA instability at low pH and recurrent failure to genetically transform acidophilic bacteria, gene transfer in acidic environments was considered negligible. Findings presented herein imply that a more or less conserved pool of actively excising MGEs occurs in the A. caldus population and point to a greater frequency of gene exchange in this econiche than previously recognized. Also, the data suggest that these elements endow the species with capacities to withstand the diverse abiotic and biotic stresses of natural

  10. The relative contribution of methanotrophs to microbial communities and carbon cycling in soil overlying a coal-bed methane seep.

    Science.gov (United States)

    Mills, Christopher T; Slater, Gregory F; Dias, Robert F; Carr, Stephanie A; Reddy, Christopher M; Schmidt, Raleigh; Mandernack, Kevin W

    2013-06-01

    Seepage of coal-bed methane (CBM) through soils is a potential source of atmospheric CH4 and also a likely source of ancient (i.e. (14) C-dead) carbon to soil microbial communities. Natural abundance (13) C and (14) C compositions of bacterial membrane phospholipid fatty acids (PLFAs) and soil gas CO2 and CH4 were used to assess the incorporation of CBM-derived carbon into methanotrophs and other members of the soil microbial community. Concentrations of type I and type II methanotroph PLFA biomarkers (16:1ω8c and 18:1ω8c, respectively) were elevated in CBM-impacted soils compared with a control site. Comparison of PLFA and 16s rDNA data suggested type I and II methanotroph populations were well estimated and overestimated by their PLFA biomarkers, respectively. The δ(13) C values of PLFAs common in type I and II methanotrophs were as negative as -67‰ and consistent with the assimilation of CBM. PLFAs more indicative of nonmethanotrophic bacteria had δ(13) C values that were intermediate indicating assimilation of both plant- and CBM-derived carbon. Δ(14) C values of select PLFAs (-351 to -936‰) indicated similar patterns of CBM assimilation by methanotrophs and nonmethanotrophs and were used to estimate that 35-91% of carbon assimilated by nonmethanotrophs was derived from CBM depending on time of sampling and soil depth.

  11. The relative contribution of methanotrophs to microbial communities and carbon cycling in soil overlying a coal-bed methane seep

    Science.gov (United States)

    Mills, Christopher T.; Slater, Gregory F.; Dias, Robert F.; Carr, Stephanie A.; Reddy, Christopher M.; Schmidt, Raleigh; Mandernack, Kevin W.

    2013-01-01

    Seepage of coal-bed methane (CBM) through soils is a potential source of atmospheric CH4 and also a likely source of ancient (i.e. 14C-dead) carbon to soil microbial communities. Natural abundance 13C and 14C compositions of bacterial membrane phospholipid fatty acids (PLFAs) and soil gas CO2 and CH4 were used to assess the incorporation of CBM-derived carbon into methanotrophs and other members of the soil microbial community. Concentrations of type I and type II methanotroph PLFA biomarkers (16:1ω8c and 18:1ω8c, respectively) were elevated in CBM-impacted soils compared with a control site. Comparison of PLFA and 16s rDNA data suggested type I and II methanotroph populations were well estimated and overestimated by their PLFA biomarkers, respectively. The δ13C values of PLFAs common in type I and II methanotrophs were as negative as −67‰ and consistent with the assimilation of CBM. PLFAs more indicative of nonmethanotrophic bacteria had δ13C values that were intermediate indicating assimilation of both plant- and CBM-derived carbon. Δ14C values of select PLFAs (−351 to −936‰) indicated similar patterns of CBM assimilation by methanotrophs and nonmethanotrophs and were used to estimate that 35–91% of carbon assimilated by nonmethanotrophs was derived from CBM depending on time of sampling and soil depth.

  12. Effects of granular activated carbon on methane removal performance and methanotrophic community of a lab-scale bioreactor.

    Science.gov (United States)

    Lee, Eun-Hee; Choi, Sun-Ah; Yi, Taewoo; Kim, Tae Gwan; Lee, Sang-Don; Cho, Kyung-Suk

    2015-01-01

    Two identical lab-scale bioreactor systems were operated to examine the effects of granular activated carbon (GAC) on methane removal performance and methanotrophic community. Both bioreactor systems removed methane completely at a CH4 loading rate of 71.2 g-CH4·d(-1) for 17 days. However, the methane removal efficiency declined to 88% in the bioreactor without GAC, while the bioreactor amended with GAC showed greater methane removal efficiency of 97% at a CH4 loading rate of 107.5 g-CH4·d(-1). Although quantitative real-time PCR showed that methanotrophic populations were similar levels of 5-10 × 10(8) pmoA gene copy number·VSS(-1) in both systems, GAC addition changed the methanotrophic community composition of the bioreactor systems. Microarray assay revealed that GAC enhanced the type I methanotrophic genera including Methylobacter, Methylomicrobium, and Methylomonas of the system, which suggests that GAC probably provided a favorable environment for type I methanotrophs. These results indicated that GAC is a promising support material in bioreactor systems for CH4 mitigation.

  13. Isolation

    DEFF Research Database (Denmark)

    Agerholm, Frank Juul

    2011-01-01

    Næringsstoffet har i dette nummer sat fokus på ”velvære i vinterkulden”, ”indendørsaktiviteter” og ”fedtafgift”. I klummen vises det, at disse tre fokusområder, der for en umiddelbar betragtning måske nok synes noget uensartede, falder sammen i ét tema: Isolation!......Næringsstoffet har i dette nummer sat fokus på ”velvære i vinterkulden”, ”indendørsaktiviteter” og ”fedtafgift”. I klummen vises det, at disse tre fokusområder, der for en umiddelbar betragtning måske nok synes noget uensartede, falder sammen i ét tema: Isolation!...

  14. High Throughput Sequencing to Detect Differences in Methanotrophic Methylococcaceae and Methylocystaceae in Surface Peat, Forest Soil, and Sphagnum Moss in Cranesville Swamp Preserve, West Virginia, USA

    Directory of Open Access Journals (Sweden)

    Evan Lau

    2015-04-01

    Full Text Available Northern temperate forest soils and Sphagnum-dominated peatlands are a major source and sink of methane. In these ecosystems, methane is mainly oxidized by aerobic methanotrophic bacteria, which are typically found in aerated forest soils, surface peat, and Sphagnum moss. We contrasted methanotrophic bacterial diversity and abundances from the (i organic horizon of forest soil; (ii surface peat; and (iii submerged Sphagnum moss from Cranesville Swamp Preserve, West Virginia, using multiplex sequencing of bacterial 16S rRNA (V3 region gene amplicons. From ~1 million reads, >50,000 unique OTUs (Operational Taxonomic Units, 29 and 34 unique sequences were detected in the Methylococcaceae and Methylocystaceae, respectively, and 24 potential methanotrophs in the Beijerinckiaceae were also identified. Methylacidiphilum-like methanotrophs were not detected. Proteobacterial methanotrophic bacteria constitute <2% of microbiota in these environments, with the Methylocystaceae one to two orders of magnitude more abundant than the Methylococcaceae in all environments sampled. The Methylococcaceae are also less diverse in forest soil compared to the other two habitats. Nonmetric multidimensional scaling analyses indicated that the majority of methanotrophs from the Methylococcaceae and Methylocystaceae tend to occur in one habitat only (peat or Sphagnum moss or co-occurred in both Sphagnum moss and peat. This study provides insights into the structure of methanotrophic communities in relationship to habitat type, and suggests that peat and Sphagnum moss can influence methanotroph community structure and biogeography.

  15. Seasonal and spatial aspects of the eco-distribution of methanotrophic bacteria in floodplain soils

    Science.gov (United States)

    Bodelier, P. L. E.; Meima-Franke, M.; Kamst, M.; Bodrossy, L.; Stralis-Pavese, N.; Hefting, M. M.; Laanbroek, R.

    2009-04-01

    METHECO is the acronym of a consortium of research groups funded by the European Science foundation (ESF) within the EuroDIVERSITY program. The consortium investigates the role of microbial diversity in the dynamics and stability of global methane consumption. The consortium covers various habitats (i.e. Landfills, rice paddies, alpine meadows, littoral wetlands, forests, arctic wetlands, peat soils and river floodplains) and assesses the effects of natural environmental perturbation on the function structure relationship of methane-consuming microbial communities. Consortium members follow the same experimental and methodological scheme using DNA and RNA based techniques (i.e. pmoA-based cloning, DGGE, micro arrays, Real Time PCR, stable isotope probing). This paper presents the results obtained in a river floodplain along the river Rhine in the Netherlands, a habitat anticipated to be subjected to major changes in flooding regime due to climate change. Experiments were carried out to assess methanotrophic diversity, methane oxidation kinetics and spatial variability of function and structure of methane-oxidizing communities. Flooding events affected methane consumption negatively on short term. However, the long -term consequences of the flooding regime where the establishment of a distinct maximum methane consumption activity exactly in the part of the floodplain intermediate between permanently and irregularly flooded, where moisture and organic matter content were optimal for methane cycling. The methanotrophic community composition as analysed by pmoA micro array mirrored the result of the activity measurements, demonstrating that the communities differed clearly according to the flooding gradient. Diversity as assessed by micro array and activity components (initial consumption, Vmax, Vmax/Km) were positively correlated. QPCR analyses showed that main types of methanotrophic bacteria were differentially distributed throughout the flooding gradient. Type I

  16. Geochemical niches of iron-oxidizing acidophiles in acidic coal mine drainage.

    Science.gov (United States)

    Jones, Daniel S; Kohl, Courtney; Grettenberger, Christen; Larson, Lance N; Burgos, William D; Macaladya, Jennifer L

    2015-02-01

    A legacy of coal mining in the Appalachians has provided a unique opportunity to study the ecological niches of iron-oxidizing microorganisms. Mine-impacted, anoxic groundwater with high dissolved-metal concentrations emerges at springs and seeps associated with iron oxide mounds and deposits. These deposits are colonized by iron-oxidizing microorganisms that in some cases efficiently remove most of the dissolved iron at low pH, making subsequent treatment of the polluted stream water less expensive. We used full-cycle rRNA methods to describe the composition of sediment communities at two geochemically similar acidic discharges, Upper and Lower Red Eyes in Somerset County, PA, USA. The dominant microorganisms at both discharges were acidophilic Gallionella-like organisms, “Ferrovum” spp., and Acidithiobacillus spp. Archaea and Leptospirillum spp. accounted for less than 2% of cells. The distribution of microorganisms at the two sites could be best explained by a combination of iron(II) concentration and pH. Populations of the Gallionella-like organisms were restricted to locations with pH>3 and iron(II) concentration of >4 mM, while Acidithiobacillus spp. were restricted to pHiron(II) concentration of iron(II) concentration of >4 mM. Our findings offer a predictive framework that could prove useful for describing the distribution of microorganisms in acid mine drainage, based on readily accessible geochemical parameters.

  17. Solar Radiation Stress in Natural Acidophilic Biofilms of Euglena mutabilis Revealed by Metatranscriptomics and PAM Fluorometry.

    Science.gov (United States)

    Puente-Sánchez, Fernando; Olsson, Sanna; Gómez-Rodriguez, Manuel; Souza-Egipsy, Virginia; Altamirano-Jeschke, Maria; Amils, Ricardo; Parro, Victor; Aguilera, Angeles

    2016-02-01

    The daily photosynthetic performance of a natural biofilm of the extreme acidophilic Euglena mutabilis from Río Tinto (SW, Spain) under full solar radiation was analyzed by means of pulse amplitude-modulated (PAM) fluorescence measurements and metatrascriptomic analysis. Natural E. mutabilis biofilms undergo large-scale transcriptomic reprogramming during midday due to a dynamic photoinhibition and solar radiation stress. Photoinhibition is due to UV radiation and not to light intensity, as revealed by PAM fluorometry analysis. In order to minimize the negative effects of solar radiation, our data supports the presence of a circadian rhythm in this euglenophyte that increases their opportunity to survive. Differential gene expression throughout the day (at 12:00, 20:00 and night) was monitored by massive Illumina parallel sequencing of metatranscriptomic libraries. The transcription pattern was altered in genes involved in Photosystem II stability and repair, UV damaged DNA repair, non-photochemical quenching and oxidative stress, supporting the photoinhibition detected by PAM fluorometry at midday.

  18. Biodiversity and interactions of acidophiles: Key to understanding and optimizing microbial processing of ores and concentrates

    Institute of Scientific and Technical Information of China (English)

    D.B.JOHNSON

    2008-01-01

    Mining companies have become increasingly aware of the potential of microbiological approaches for recovering base and precious metals from low-grade ores,and for remediating acidic,metal-rich wastewaters that drain from both operating and abandoned mine sites.Biological systems offer a number of environmental and (sometimes) economical advantages over conventional approaches,such as pyrometallurgy,though their application is not appropriate in every situation.Mineral processing using micro-organisms has been exploited for extracting gold,copper,uranium and cobalt,and current developments are targeting other base metals.Recently,there has been a great increase in our knowledge and understanding of both the diversity of the microbiology of biomining environments,and of how the microorganisms interact with each other.The results from laboratory experiments which have simulated both stirred tank and heap bioreactor systems have shown that microbial consortia are more robust than pure cultures of mineral-oxidizing acidophiles,and also tend to be more effective at bioleaching and bio-oxidizing ores and concentrates.The paper presented a concise review of the nature and interactions of microbial consortia that are involved in the oxidation of sulfide minerals,and how these might be adapted to meet future challenges in biomining operations.

  19. Construction of the astaxanthin biosynthetic pathway in a methanotrophic bacterium Methylomonas sp. strain 16a.

    Science.gov (United States)

    Ye, Rick W; Yao, Henry; Stead, Kristen; Wang, Tao; Tao, Luan; Cheng, Qiong; Sharpe, Pamela L; Suh, Wonchul; Nagel, Eva; Arcilla, Dennis; Dragotta, Dominic; Miller, Edward S

    2007-04-01

    Methylomonas sp. strain 16a is an obligate methanotrophic bacterium that uses methane or methanol as the sole carbon source. An effort was made to engineer this organism for astaxanthin production. Upon expressing the canthaxanthin gene cluster under the control of the native hps promoter in the chromosome, canthaxanthin was produced as the main carotenoid. Further conversion to astaxanthin was carried out by expressing different combinations of crtW and crtZ genes encoding the beta-carotenoid ketolase and hydroxylase. The carotenoid intermediate profile was influenced by the copy number of these two genes under the control of the hps promoter. Expression of two copies of crtZ and one copy of crtW led to the accumulation of a large amount of the mono-ketolated product adonixanthin. On the other hand, expression of two copies of crtW and one copy of crtZ resulted in the presence of non-hydroxylated carotenoid canthaxanthin and the mono-hydroxylated adonirubin. Production of astaxanthin as the predominant carotenoid was obtained in a strain containing two complete sets of carotenoid biosynthetic genes. This strain had an astaxanthin titer ranging from 1 to 2.4 mg g(-1) of dry cell biomass depending on the growth conditions. More than 90% of the total carotenoid was astaxanthin, of which the majority was in the form of E-isomer. This result indicates that it is possible to produce astaxanthin with desirable properties in methanotrophs through genetic engineering.

  20. In situ bioremediation of trichloroethylene-contaminated water by a resting-cell methanotrophic microbial filter

    Energy Technology Data Exchange (ETDEWEB)

    Taylor, R T; Duba, A G; Durham, W B; Hanna, M L; Jackson, K J; Jovanovich, M C; Knapp, R B; Knezovich, J P; Shah, N N; Shonnard, D R; Wijesinghe, A M

    1992-10-01

    The Lawrence Livermore National Laboratory is testing and developing an in situ microbial filter technology for remediating migrating subsurface plumes contaminated with low concentrations of trichloroethylene (TCE). Their current focus is the establishment of a replenishable bioactive zone (catalytic filter) along expanding plume boundaries by the Injection of a representative methanotrophic bacterium, Methylosinus trichosporium OB3b. We have successfully demonstrated this microbial filter strategy using emplaced, attached resting cells (no methane additions) in a 1.1-m flow-through test bed loaded with water-saturated sand. Two separate 24 h pulses of TCE (109 ppb and 85 ppb), one week apart, were pumped through the system at a flow velocity of 1.5 cm/h; no TCE (<0.5 ppb) was detected on the downstream side of the microbial filter. Subsequent excavation of the wet sand confirmed the existence of a TCE-bioactive zone 19 days after it had been created. An enhanced longevity of the cellular, soluble-form methane monooxygenase produced by this methanotroph Is a result of our laboratory bioreactor culturing conditions. Additional experiments with cells in sealed vials and emplaced in the 1.1-m test bed yielded a high resting-cell finite TCE biotransformation capacity of [approximately] 0.25 mg per mg of bacteria; this is suitable for a planned sand-filled trench field demonstration at a Lawrence Livermore National Laboratory site.

  1. Biosynthesis of poly-3-hydroxybutyrate with a high molecular weight by methanotroph from methane and methanol

    Institute of Scientific and Technical Information of China (English)

    Yingxin Zhang; Jiaying Xin; Linlin Chen; Hao Song; Chungu Xia

    2008-01-01

    Poly-3-hydroxybutyrate (PHB) can be produced by various species of bacteria. Among the possible carbon sources, both methane and methanol could be a suitable substrate for the production of PHB. Methane is cheap and plentiful not only as natural gas, but also as biogas. Methanol can also maintain methanotrophic activity in some conditions. The methanotrophic strain Methylosinus trichosporium IMV3011 can accumulate PHB with methane and methanol in a brief nonsterile process. Liquid methanol (0.1%) was added to improve the oxidization of methane. The studies were carried out using shake flasks. Cultivation was performed in two stages: a continuous growth phase and a PHB accumulation phase under the conditions short of essential nutrients (ammonium, nitrate, phosphorus, copper, iron (III), magnesium or ethylenediamine tetraacetate (EDTA)) in batch culture. It was found that the most suitable growth time for the cell is 144 h. Then an optimized culture condition for second stage was determined, in which the PHB concentration could be much increased to 0.6 g/L. In order to increase PHB content, citric acid was added as an inhibitor of tricarboxylic acid cycle (TCA). It was found that citric acid is favorable for the PHB accumulation, and the PHB yield was increased to 40% (w/w) from the initial yield of 12% (w/w) after nutrient deficiency cultivation. The PHB produced is of very high quality with molecular weight up to 1.5 ×106Da.

  2. Effect of nitrogen fertilization on methane oxidation, abundance, community structure, and gene expression of methanotrophs in the rice rhizosphere.

    Science.gov (United States)

    Shrestha, Minita; Shrestha, Pravin Malla; Frenzel, Peter; Conrad, Ralf

    2010-12-01

    Nitrogen, one of the limiting factors for the yield of rice, can also have an important function in methane oxidation, thus affecting its global budget. Rice microcosms, planted in the greenhouse, were treated with the N-fertilizers urea (UPK) and ammonium sulfate (APK) or were only treated with phosphorous and potassium (PK). Methane oxidation rates in PK and UPK treatments were similar during most of the rice-growing season, revealing no effect of urea. However, ammonium sulfate strongly suppressed methanogenesis providing an unfavorable environment for methanotrophs in APK treatment. Roots and rhizospheric soil samples, collected from six different growth stages of the rice plant, were analyzed by terminal restriction fragment length polymorphism (T-RFLP) of the pmoA gene. Assignment of abundant T-RFs to cloned pmoA sequences indicated that the populations on roots were dominated by type-I methanotrophs, whereas the populations in rhizospheric soil were dominated by type-II methanotrophs irrespectively of growth stages and fertilizer treatments. Non-metric multidimensional scaling ordination analysis of T-RFLP profiles revealed that the methanotrophic community was significantly (PAPK treatment.

  3. Autotrophic Carbon Dioxide Fixation via the Calvin-Benson-Bassham Cycle by the Denitrifying Methanotroph "Candidatus Methylomirabilis oxyfera"

    NARCIS (Netherlands)

    Rasigraf, O.; Kool, D.M.; Jetten, M.S.M.; Ettwig, K.F.; Sinninghe Damsté, J.S.

    2014-01-01

    Methane is an important greenhouse gas and the most abundant hydrocarbon in the Earth's atmosphere. Methanotrophic microorganisms can use methane as their sole energy source and play a crucial role in the mitigation of methane emissions in the environment. "Candidatus Methylomirabilis oxyfera" is a

  4. Spatial Patterns of Soil Development, Methane Oxidation, and Methanotrophic Diversity along a Receding Glacier Forefield, Southeast Greenland

    DEFF Research Database (Denmark)

    Barcena, Teresa Gomez; Finster, Kai; Yde, Jacob Clement

    2011-01-01

    .24 nmol CH4 day−1 gsoil−1 at 10 °C in the LIA terminal moraine. Methane consumption was not detected in younger samples, despite the presence of high-affinity methanotrophs in all samples. This was indicated by successful amplification of partial pmoA genes, which code for a subunit of a key enzyme...

  5. Hydrology is reflected in the functioning and community composition of methanotrophs in the littoral wetland of a boreal lake

    NARCIS (Netherlands)

    Siljanen, H.M.P.; Saari, A.; Krause, S.; Lensu, A.; Abell, G.C.J.; Bodrossy, L.; Bodelier, P.L.E.; Martikainen, P.J.

    2011-01-01

    In lake ecosystems a major proportion of methane emissions originate from the littoral zone which can have a great spatial variability in hydrology, soil quality and vegetation. Hitherto, spatial heterogeneity and the effects it has on functioning and diversity of methanotrophs in littoral wetlands

  6. Production of uracil from methane by a newly isolated Methylomonas sp. SW1.

    Science.gov (United States)

    Kim, Sangwoo; Lee, Wangjun; Song, Insu; Kwon, Yuhyun; Yun, Seokhun; Park, Soohyun; Cho, Sukhyeong; Oh, Byung-Keun; Oh, Han Bin; Lee, Jinwon

    2016-12-20

    Methane is an abundant, inexpensive one-carbon feedstock and one of the most powerful greenhouse gases. Because it does not compete with food demand, it is considered a promising carbon feedstock for the production of valuable products using methanotrophic bacteria. Here, we isolated a novel methanotrophic bacterium, Methylomonas sp. SW1, from a sewage sample obtained from Wonju City Water Supply Drainage Center, Republic of Korea. The conditions for uracil production by Methylomonas sp. SW1, such as Cu(2+) concentration and temperature were investigated and optimized. As a result, Methylomonas sp. SW1 produced uracil from methane as a sole carbon source with a titer of 2.1mg/L in 84h without genetic engineering under the optimized condition. The results in this study demonstrate the feasibility of using Methylomonas sp. SW1 for the production of uracil from methane. This is the first report of uracil production from gas feedstock by methanotrophic bacteria.

  7. Warmer and drier conditions and nitrogen fertilizer application altered methanotroph abundance and methane emissions in a vegetable soil.

    Science.gov (United States)

    Ran, Yu; Xie, Jianli; Xu, Xiaoya; Li, Yong; Liu, Yapeng; Zhang, Qichun; Li, Zheng; Xu, Jianming; Di, Hongjie

    2016-11-12

    Methane (CH4) is a potent greenhouse gas, and soil can both be a source and sink for atmospheric CH4. It is not clear how future climate change may affect soil CH4 emissions and related microbial communities. The aim of this study was to determine the interactive effects of a simulated warmer and drier climate scenarios and the application of different nitrogen (N) sources (urea and manure) on CH4 emissions and related microbial community abundance in a vegetable soil. Greenhouses were used to control simulated climate conditions which gave 2.99 °C warmer and 6.2% lower water content conditions. The field experiment was divided into two phases. At the beginning of phase II, half of the greenhouses were removed to study possible legacy effects of the simulated warmer and drier conditions. The responses in methanogen and methanotroph abundance to a simulated climate change scenario were determined using real-time PCR. The results showed that the simulated warmer and drier conditions in the greenhouses significantly decreased CH4 emissions largely due to the lower soil moisture content. For the same reason, CH4 emissions of treatments in phase I were much lower than the same treatments in phase II. The abundance of methanotrophs showed a more significant response than methanogens to the simulated climate change scenario, increasing under simulated drier conditions. Methanogenic community abundance remained low, except where manure was applied which provided a source of organic C that stimulated methanogen growth. Soil moisture content was a major driver for methanotroph abundance and strongly affected CH4 emissions. The application of N source decreased CH4 emissions probably because of increased methanotrophic activity. CH4 emissions were positively correlated to methanogenic abundance and negatively correlated to methanotrophic abundance. These results demonstrate that projected future climate change conditions can have a feedback impact on CH4 emissions from the

  8. Autotrophic carbon dioxide fixation via the Calvin-Benson-Bassham cycle by the denitrifying methanotroph "Candidatus Methylomirabilis oxyfera".

    Science.gov (United States)

    Rasigraf, Olivia; Kool, Dorien M; Jetten, Mike S M; Sinninghe Damsté, Jaap S; Ettwig, Katharina F

    2014-04-01

    Methane is an important greenhouse gas and the most abundant hydrocarbon in the Earth's atmosphere. Methanotrophic microorganisms can use methane as their sole energy source and play a crucial role in the mitigation of methane emissions in the environment. "Candidatus Methylomirabilis oxyfera" is a recently described intra-aerobic methanotroph that is assumed to use nitric oxide to generate internal oxygen to oxidize methane via the conventional aerobic pathway, including the monooxygenase reaction. Previous genome analysis has suggested that, like the verrucomicrobial methanotrophs, "Ca. Methylomirabilis oxyfera" encodes and transcribes genes for the Calvin-Benson-Bassham (CBB) cycle for carbon assimilation. Here we provide multiple independent lines of evidence for autotrophic carbon dioxide fixation by "Ca. Methylomirabilis oxyfera" via the CBB cycle. The activity of ribulose-1,5-bisphosphate carboxylase/oxygenase (RubisCO), a key enzyme of the CBB cycle, in cell extracts from an "Ca. Methylomirabilis oxyfera" enrichment culture was shown to account for up to 10% of the total methane oxidation activity. Labeling studies with whole cells in batch incubations supplied with either (13)CH4 or [(13)C]bicarbonate revealed that "Ca. Methylomirabilis oxyfera" biomass and lipids became significantly more enriched in (13)C after incubation with (13)C-labeled bicarbonate (and unlabeled methane) than after incubation with (13)C-labeled methane (and unlabeled bicarbonate), providing evidence for autotrophic carbon dioxide fixation. Besides this experimental approach, detailed genomic and transcriptomic analysis demonstrated an operational CBB cycle in "Ca. Methylomirabilis oxyfera." Altogether, these results show that the CBB cycle is active and plays a major role in carbon assimilation by "Ca. Methylomirabilis oxyfera" bacteria. Our results suggest that autotrophy might be more widespread among methanotrophs than was previously assumed and implies that a methanotrophic

  9. Unexpected DNA-fingerprinting pattern in a deep peat bog: evidence for methanotrophs at the bottom?

    Science.gov (United States)

    Steinmann, P.; Rossi, P.; Huon, S.; Eilrich, B.; Casati, S.

    2003-04-01

    With the goal of a better understanding of the fate of methane in the deep layers of peat bogs, we analysed the microbial 16S rDNA gene pool and measured the stable carbon isotope composition of bulk peat of a deep (6 m) peat bog profile (Etang de la Gruyère, Switzerland). Both Bacterial and Archaean communities were assessed using respectively TTGE (Temporal Temperature Gradient Electrophoresis) and SSCP (Single Strand Conformation Polymorphism), with fragments of the V1-V3 region of the 16S rDNA gene. The "relative diversity" shown in the TTGE AND SSCP gel patterns is presented using indices and band numbers per sample (Simpson evenness). PCA was calculated on the basis of the intensities of all bands found in the TTGE and SSCP fingerprinting profiles. These DNA fingerprinting patterns reveal the presence of a structured microbial community throughout the whole depth profile. Clear differences can be observed between the communities found in the near surface layers and those found at depth. Surprisingly, for both Archaean and Bacterial communities, the deepest samples display a high similarity level with those found in the first 20 centimeters. The δ13C values of the peat are relatively constant from the surface of the bog down to a depth of 5 m (values between 25.5 ppm and 26.5 ppm). Below 5 m the values decrease considerably with depth ( 28.5 ppm). As a working hypothesis to explain the two observations, we consider the possibility of the presence of methanotrophs in the deepest parts of the bogs. The electron acceptors needed for methane oxidation could be derived from lateral advection of less reducing groundwater. However, available pore water analyses suggest that neither molecular oxygen, nor sulfate or nitrate are present. One possible oxidising agent would be trivalent iron (solid or colloidal). Indeed are the iron concentrations in the deeper pore waters are elevated. Such deep methanotrophic microbial community could be similar to those found near

  10. Biochemistry and Ecology of Novel Cytochromes Catalyzing Fe(II) Oxidation by an Acidophilic Microbial Community

    Science.gov (United States)

    Singer, S. W.; Jeans, C. J.; Thelen, M. P.; Verberkmoes, N. C.; Hettich, R. C.; Chan, C. S.; Banfield, J. F.

    2007-12-01

    An acidophilic microbial community found in the Richmond Mine at Iron Mountain, CA forms abundant biofilms in extremely acidic (pHcytochromes with unique properties. Sulfuric acid extraction of biofilm samples liberated one of these proteins, a 16 kDa cytochrome with an unusual alpha-band absorption at 579 (Cyt579). Genomic sequencing of multiple biofilms indicated that several variants of Cyt579 were present in Leptospirillum strains. Intact protein MS analysis identified the dominant variants in each biofilm and documented multiple N-terminal cleavage sites for Cyt579. By combining biochemical, geochemical and microbiological data, we established that the sequence variation and N-terminal processing of Cyt579 are selected by ecological conditions. In addition to the soluble Cyt579, the second cytochrome appears as a much larger protein complex of ~210 kDa predominant in the biofilm membrane fraction, and has an alpha-band absorption at 572 nm. The 60 kDa cytochrome subunit, Cyt572, resides in the outer membrane of LeptoII, and readily oxidizes Fe(II) at low pH (0.95 - 3.0). Several genes encoding Cyt572 were localized within a recombination hotspot between two strains of LeptoII, causing a large range of variation in the sequences. Genomic sequencing and MS proteomic studies established that the variants were also selected by ecological conditions. A general mechanistic model for Fe(II) oxidation has been developed from these studies. Initial Fe(II) oxidation by Cyt572 occurs at the outer membrane. Cyt572 then transfers electrons to Cyt579, perhaps representing an initial step in energy flow to the biofilm community. Amino acid variations and post-translational modifications of these unique cytochromes may represent fine-tuning of function in response to local environmental conditions.

  11. Arsenic oxidation and bioaccumulation by the acidophilic protozoan, Euglena mutabilis, in acid mine drainage (Carnoules, France)

    Energy Technology Data Exchange (ETDEWEB)

    Casiot, C.; Bruneel, O.; Personne, J.-C.; Leblanc, M.; Elbaz-Poulichet, F. [University of Montpellier 2, Montpellier (France)

    2004-03-29

    In the acid stream (pH 2.5-4.7) originating from the Camoules mine tailings, the acidophilic protozoan Euglena mutabilis grows with extremely high sulfate (1.9-4.9 g/l), iron (0.7-1.7 g/l) and arsenic concentrations (0.08-0.26 g/l). Strong variations in flow rate and high sulfate concentrations (up to 4.9 g/l) have been registered in early winter and might be the reason for the reduction in cell number of the protozoan from October to December 2001. No relation was established between arsenic concentration and/or speciation and abundance of the protozoan in the stream. Arsenite, which is the most toxic form, predominates in water. The oxidation of arsenite to arsenate occurred within a few days in laboratory experiments when E. mutabilis was present in Reigous Creek water and synthetic As(III)-rich culture medium. Methylated compounds (MMA, DMA) were not identified in the culture media. The protozoan bioaccumulated As in the cell (336{+-} 112 {mu}g As/g dry wt.) as inorganic arsenite (105 {+-} 52 {mu}g As/g dry wt.) and arsenate (231 {+-} 112 {mu}g As/g dry wt.). Adsorption of As at the cell surface reached 57 mg/g dry wt. in the As(V) form for E. mutabilis grown in 250 mg/l As(III) synthetic medium. Both intracellular accumulation and adsorption at the cell surface increased for increasing As(III) concentration in the medium but the concentration factor in the cell relative to soluble As decreased.

  12. Microalgae as a safe food source for animals: nutritional characteristics of the acidophilic microalga Coccomyxa onubensis

    Directory of Open Access Journals (Sweden)

    Francisco Navarro

    2016-10-01

    Full Text Available Background: Edible microalgae are marine or fresh water mesophilic species. Although the harvesting of microalgae offers an abundance of opportunities to the food and pharmaceutical industries, the possibility to use extremophilic microalgae as a food source for animals is not well-documented. Objective: We studied the effects of dietary supplementation of a powdered form of the acidophilic microalga Coccomyxa onubensis on growth and health parameters of laboratory rats. Method: Four randomly organized groups of rats (n=6 were fed a standard diet (Diet 1, control or with a diet in which 0.4% (Diet 2, 1.25% (Diet 3, or 6.25% (Diet 4 (w/w of the standard diet weight was substituted with dried microalgae powder, respectively. The four groups of animals were provided ad libitum access to feed for 45 days. Results: C. onubensis biomass is rich in protein (44.60% of dry weight and dietary fiber (15.73%, and has a moderate carbohydrate content (24.8% and a low lipid content (5.4% in which polyunsaturated fatty acids represent 65% of the total fatty acid. Nucleic acids are present at 4.8%. No significant difference was found in growth rates or feed efficiency ratios of the four groups of rats. Histological studies of liver and kidney tissue revealed healthy organs in control and C. onubensis-fed animals, while plasma hematological and biochemical parameters were within healthy ranges for all animals. Furthermore, animals fed a microalgae-enriched diet exhibited a statistically significant decrease in both blood cholesterol and triglyceride levels. The blood triglyceride content and very low density lipoprotein-cholesterol levels decreased by about 50% in rats fed Diet 4. Conclusions: These data suggest that C. onubensis may be useful as a food supplement for laboratory animals and may also serve as a nutraceutical in functional foods. In addition, microalgae powder-supplemented diets exerted a significant hypocholesterolemic and hypotriglyceridemic

  13. Aerobic methanotrophic communities at the Red Sea brine-seawater interface

    Directory of Open Access Journals (Sweden)

    Rehab Z. Abdallah

    2014-09-01

    Full Text Available The central rift of the Red Sea contains 25 brine pools with different physicochemical conditions, dictating the diversity and abundance of the microbial community. Three of these pools, the Atlantis II, Kebrit and Discovery Deeps, are uniquely characterized by a high concentration of hydrocarbons. The brine-seawater interface, described as an anoxic-oxic (brine-seawater boundary, is characterized by a high methane concentration, thus favoring aerobic methane oxidation. The current study analyzed the aerobic free–living methane-oxidizing bacterial communities that potentially contribute to methane oxidation at the brine-seawater interfaces of the three aforementioned brine pools, using metagenomic pyrosequencing, 16S rRNA pyrotags and pmoA library constructs. The sequencing of 16S rRNA pyrotags revealed that these interfaces are characterized by high microbial community diversity. Signatures of aerobic methane-oxidizing bacteria were detected in the Atlantis II Interface (ATII-I and the Kebrit Deep Upper (KB-U and Lower (KB-L brine-seawater interfaces. Through phylogenetic analysis of pmoA, we further demonstrated that the ATII-I aerobic methanotroph community is highly diverse. We propose four ATII-I pmoA clusters. Most importantly, cluster 2 groups with marine methane seep methanotrophs, and cluster 4 represent a unique lineage of an uncultured bacterium with divergent alkane monooxygenases. Moreover, non-metric multidimensional scaling (NMDS based on the ordination of putative enzymes involved in methane metabolism showed that the Kebrit interface layers were distinct from the ATII-I and DD-I brine-seawater interfaces.

  14. Study on processing highly effective Acidophilic bacteria starter culture and Acidophilic bacteria milk powder with new technology%新工艺制备高效嗜酸菌发酵剂和嗜酸菌奶粉的研究

    Institute of Scientific and Technical Information of China (English)

    方景泉; 徐欣

    2013-01-01

    Two different strains of acidophilic bacteria were incubated by different methods and through a series contrast tests,we found the best method for incubating.Then concentrated the bacteria with high sPeed centrifuge and put it in 10% skim milk solution with protective agent D,after vacuum freeze drying the viable count of Lactobacillus acidophilus exist in the starter culture were 8.3 ×1010g-1and its survival rate could be 68.8%.We can process Acidophilic bacteria milk powder when mixed it together with milk powder.%分别使两株嗜酸菌在脱脂乳水解液培养基和脱脂乳培养基中生长,通过一系列的对比实验,以及使菌体浓缩,浓菌体悬浮在添加保护剂D质量分数为10%脱脂乳中,经冷冻干燥后,所得高效嗜酸菌发酵剂中的细胞数达8.3×1010 g-1.其在真空冷冻干燥后的存活率最高可达68.8%.将此高效发酵剂与奶粉混合即可制得嗜酸菌奶粉.

  15. Investigation of energy gene expressions and community structures of free and attached acidophilic bacteria in chalcopyrite bioleaching.

    Science.gov (United States)

    Zhu, Jianyu; Jiao, Weifeng; Li, Qian; Liu, Xueduan; Qin, Wenqing; Qiu, Guanzhou; Hu, Yuehua; Chai, Liyuan

    2012-12-01

    In order to better understand the bioleaching mechanism, expression of genes involved in energy conservation and community structure of free and attached acidophilic bacteria in chalcopyrite bioleaching were investigated. Using quantitative real-time PCR, we studied the expression of genes involved in energy conservation in free and attached Acidithiobacillus ferrooxidans during bioleaching of chalcopyrite. Sulfur oxidation genes of attached A. ferrooxidans were up-regulated while ferrous iron oxidation genes were down-regulated compared with free A. ferrooxidans in the solution. The up-regulation may be induced by elemental sulfur on the mineral surface. This conclusion was supported by the results of HPLC analysis. Sulfur-oxidizing Acidithiobacillus thiooxidans and ferrous-oxidizing Leptospirillum ferrooxidans were the members of the mixed culture in chalcopyrite bioleaching. Study of the community structure of free and attached bacteria showed that A. thiooxidans dominated the attached bacteria while L. ferrooxidans dominated the free bacteria. With respect to available energy sources during bioleaching of chalcopyrite, sulfur-oxidizers tend to be on the mineral surfaces whereas ferrous iron-oxidizers tend to be suspended in the aqueous phase. Taken together, these results indicate that the main role of attached acidophilic bacteria was to oxidize elemental sulfur and dissolution of chalcopyrite involved chiefly an indirect bioleaching mechanism.

  16. The acidophilic microorganisms diversity present in lignite and pit coal from Paroseni, Halânga, Turceni mines

    Directory of Open Access Journals (Sweden)

    Carmen Madalina CISMASIU

    2009-11-01

    Full Text Available Pollution from coal combustion is the largest problem in the current use of coal and the biggest constraint on the increased use of coal. When these fossil fuels are combusted, sulphur-di-oxide is released into the atmosphere causing acid rains which dissolves buildings, kills forest. Knowing the physiological groups of microorganisms present in the coal samples has an ecological importance, completing the knowledge in the field of the microorganism’s ecology and a practical importance, being a source of new microorganisms with biotechnological potential. The microbial communities evidenced in such sites include both groups of chemolithotrophic microorganisms involved in the metals biosolubilization processes and groups of heterotrophic microorganisms involved in the processes of bioaccumulation or biofixation of metallic ions. In this context, this paper presents the study regarding the main physiological groups of microorganisms present in the pit coal and lignite samples after the industrial processing of coal. The results revealed that the microorganisms belonging to the following physiological groups: aerobic heterotrophic acidophilic bacteria, strictly anaerobic heterotrophic (sulphur-reducing, nitrifying bacteria (nitrite and nitrate bacteria, denitrifying bacteria and acidophilic chemolithotrophic bacteria on Fe2+, on S0 and on S2O3.

  17. Microbial ecology on the microcosm level: Activity and population dynamics of methanotrophic bacteria during early succession in a flooded rice field soil

    Science.gov (United States)

    Krause, S.; Frenzel, P.

    2009-04-01

    Methane oxidizing bacteria (methanotrophs) play an important role in natural wetlands and rice fields preventing large amounts of methane from escaping into the atmosphere. The occurrence of both type I and type II methanotrophs in the soil surface layer has been demonstrated in many studies. However, there is no profound understanding which of them are responsible for the oxidizing activity and how they differ ecologically. Hence, a gradient microcosm system was applied simulating oxic-anoxic interfaces of water saturated soils to unravel population dynamics in early succession of methanotrophs in a flooded rice paddy. Additionally, environmental parameters were analyzed to link environment, populations, and their specific activity. We measured pmoA-based (particulate methane monooxygenase) terminal restriction fragment length polymorphism (T-RFLP) profiles both on transcription and population level. DNA T-RFLP patterns showed no major differences in the methanotrophic community structure remaining relatively constant over time. In contrast the active methanotrophic community structure as detected by pmoA mRNA T-RFLP analysis clearly demonstrated a distinct pattern from DNA T-RFLP profiles. While type II represented the most prominent group on the population level it seems to play a minor role on the transcription level. Furthermore there were no clear implications towards a link between soil parameters (e.g. NH4+ concentration) and methanotrophic community structure.

  18. Methane monooxygenase from Methylosinus trichosporium OB3b. Purification and properties of a three-component system with high specific activity from a type II methanotroph.

    Science.gov (United States)

    Fox, B G; Froland, W A; Dege, J E; Lipscomb, J D

    1989-06-15

    Methane monooxygenase has been purified from the Type II methanotroph Methylosinus trichosporium OB3b. As observed for methane monooxygenase isolated from Type I methanotrophs, three protein components are required: a 39.7-kDa NADH reductase containing 1 mol each of FAD and a [2Fe-2S] cluster, a 15.8-kDa protein factor termed component B that contains no metals or cofactors, and a 245-kDa hydroxylase which appears to contain an oxo- or hydroxo-bridged binuclear iron cluster. Through the use of stabilizing reagents, the hydroxylase is obtained in high yield and exhibits a specific activity 8-25-fold greater than reported for previous preparations. The component B and reductase exhibit 1.5- and 4-fold greater specific activity, respectively. Quantitation of the hydroxylase oxo-bridged cluster using EPR and Mössbauer spectroscopies reveals that the highest specific activity preparations (approximately 1700 nmol/min/mg) contain approximately 2 clusters/mol. In contrast, hydroxylase preparations exhibiting a wide range of specific activities below 500 nmol/min/mg contain approximately 1 cluster/mol on average. Efficient turnover coupled to NADH oxidation requires all three protein components. However, both alkanes and alkenes are hydroxylated by the chemically reduced hydroxylase under single turnover conditions in the absence of component B and the reductase. Neither of these components catalyzes hydroxylation individually nor do they significantly affect the yield of hydroxylated product from the chemically reduced hydroxylase. Hydroxylase reduced only to the mixed valent [Fe(II).Fe(III)] state is unreactive toward O2 and yields little hydroxylated product on single turnover. This suggests that the catalytically active species is the fully reduced form. The data presented here provide the first evidence based on catalysis that the site of the monooxygenation reaction is located on the hydroxylase. It thus appears likely that the oxo-bridged iron cluster is capable of

  19. Sorption of ferrous iron by EPS from the acidophilic bacterium Acidiphilium Sp.: A mechanism proposal

    Directory of Open Access Journals (Sweden)

    Tapia, Jaime M.

    2016-09-01

    Full Text Available The aim of this work was to assess the uptake of Fe(II by extracellular polymeric substances (EPS from the acidophilic bacterium Acidiphillium 3.2Sup(5. These EPS were extracted using EDTA. EPS of A. 3.2Sup(5 loaded in sorption tests with Fe(II, were characterized using the following experimental techniques: scanning electron microscopy (SEM with energy dispersive X-ray microanalysis (EDX, X-ray diffraction (XRD and Fourier transform infrared spectroscopy (FTIR. The experimental results indicate that EPS adsorb ferrous iron according to Freundlich model with a metal sorption uptake of K = 1.14 mg1-1/n L1/n g-1 and a sorption intensity of 1/n = 1.26. In addition, ferrous iron sorption by EPS took place by preferential interaction with the carboxyl group which promotes the formation of ferrous iron oxalates (FeC2O4. Since the interaction reaction was reversible (Log K = 0.77 ± 0.33, that means that the cation sorption can be reversed at convenience.El objetivo de este trabajo fue estudiar la absorción de Fe(II por Sustancias Poliméricas Extracelulares (SPE provenientes de la bacteria acidófila Acidiphilium 3.2Sup(5. Las SPE fueron extraídas usando EDTA. SPE de A. 3.2Sup(5 cargadas con Fe(II fueron caracterizadas usando las siguientes técnicas experimentales: microscopia electrónica de barrido (MEB con microanálisis de energía dispersiva de rayos X (EDX, difracción de rayos X (DRX, y espectroscopía infrarojo (IR con transformada de Fourier (EIRTF. Los resultados muestran que las SPE absorben Fe(II según el modelo de Freundlich con un coeficiente de sorción K = 1,14 mg1-1/n g-1 e intensidad 1/n = 1,26. La captación de Fe(II por las SPE ocurre a través de la formación de oxalatos de hierro (FeC2O4, a través de una reacción reversible (Log K = 0,77 ± 0,33, lo cual implica que el hierro captado podría recuperarse si fuera de interés.

  20. Draft genome sequence of the extremely acidophilic biomining bacterium Acidithiobacillus thiooxidans ATCC 19377 provides insights into the evolution of the Acidithiobacillus genus.

    Science.gov (United States)

    Valdes, Jorge; Ossandon, Francisco; Quatrini, Raquel; Dopson, Mark; Holmes, David S

    2011-12-01

    Acidithiobacillus thiooxidans is a mesophilic, extremely acidophilic, chemolithoautotrophic gammaproteobacterium that derives energy from the oxidation of sulfur and inorganic sulfur compounds. Here we present the draft genome sequence of A. thiooxidans ATCC 19377, which has allowed the identification of genes for survival and colonization of extremely acidic environments.

  1. Draft Genome Sequence of the Extremely Acidophilic Biomining Bacterium Acidithiobacillus thiooxidans ATCC 19377 Provides Insights into the Evolution of the Acidithiobacillus Genus

    OpenAIRE

    Valdes, Jorge; Ossandon, Francisco; Quatrini, Raquel; Dopson, Mark; Holmes, David S

    2011-01-01

    Acidithiobacillus thiooxidans is a mesophilic, extremely acidophilic, chemolithoautotrophic gammaproteobacterium that derives energy from the oxidation of sulfur and inorganic sulfur compounds. Here we present the draft genome sequence of A. thiooxidans ATCC 19377, which has allowed the identification of genes for survival and colonization of extremely acidic environments.

  2. Expanding the range of polyhydroxyalkanoates synthesized by methanotrophic bacteria through the utilization of omega-hydroxyalkanoate co-substrates.

    Science.gov (United States)

    Myung, Jaewook; Flanagan, James C A; Waymouth, Robert M; Criddle, Craig S

    2017-12-01

    The first methanotrophic syntheses of polyhydroxyalkanoates (PHAs) that contain repeating units beyond 3-hydroxybutyrate and 3-hydroxyvalerate are reported. New PHAs synthesized by methanotrophs include poly(3-hydroxybutyrate-co-4-hydroxybutyrate) (P(3HB-co-4HB)), poly(3-hydroxybutyrate-co-5-hydroxyvalerate-co-3-hydroxyvalerate) (P(3HB-co-5HV-co-3HV)), and poly(3-hydroxybutyrate-co-6-hydroxyhexanoate-co-4-hydroxybutyrate) (P(3HB-co-6HHx-co-4HB)). This was achieved from a pure culture of Methylocystis parvus OBBP where the primary substrate is methane and the corresponding ω-hydroxyalkanoate monomers are added as a co-substrate after the cells are subjected to nitrogen-limited conditions.

  3. Genomic reconstruction of an uncultured hydrothermal vent gammaproteobacterial methanotroph (family Methylothermaceae indicates multiple adaptations to oxygen limitation

    Directory of Open Access Journals (Sweden)

    Connor Tobias Skennerton

    2015-12-01

    Full Text Available Hydrothermal vents are an important contributor to marine biogeochemistry, producing large volumes of reduced fluids, gasses, and metals and housing unique, productive microbial and animal communities fueled by chemosynthesis. Methane is a common constituent of hydrothermal vent fluid and is frequently consumed at vent sites by methanotrophic bacteria that serve to control escape of this greenhouse gas into the atmosphere. Despite their ecological and geochemical importance, little is known about the ecophysiology of uncultured hydrothermal vent-associated methanotrophic bacteria. Using metagenomic binning techniques, we recovered and analyzed a near-complete genome from a novel gammaproteobacterial methanotroph (B42 associated with a white smoker chimney in the Southern Lau basin. B42 was the dominant methanotroph in the community, at ~80x coverage, with only four others detected in the metagenome, all on low coverage contigs (7x - 12x. Phylogenetic placement of B42 showed it is a member of the Methylothermaceae, a family currently represented by only one sequenced genome. Metabolic inferences based on the presence of known pathways in the genome showed that B42 possesses a branched respiratory chain with A- and B-family heme copper oxidases, cytochrome bd oxidase and a partial denitrification pathway. These genes could allow B42 to respire over a wide range of oxygen concentrations within the highly dynamic vent environment. Phylogenies of the denitrification genes revealed they are the result of separate horizontal gene transfer from other proteobacteria and suggest that denitrification is a selective advantage in conditions where extremely low oxygen concentrations require all oxygen to be used for methane activation.

  4. Genomic Reconstruction of an Uncultured Hydrothermal Vent Gammaproteobacterial Methanotroph (Family Methylothermaceae) Indicates Multiple Adaptations to Oxygen Limitation

    Science.gov (United States)

    Skennerton, Connor T.; Ward, Lewis M.; Michel, Alice; Metcalfe, Kyle; Valiente, Chanel; Mullin, Sean; Chan, Ken Y.; Gradinaru, Viviana; Orphan, Victoria J.

    2015-01-01

    Hydrothermal vents are an important contributor to marine biogeochemistry, producing large volumes of reduced fluids, gasses, and metals and housing unique, productive microbial and animal communities fueled by chemosynthesis. Methane is a common constituent of hydrothermal vent fluid and is frequently consumed at vent sites by methanotrophic bacteria that serve to control escape of this greenhouse gas into the atmosphere. Despite their ecological and geochemical importance, little is known about the ecophysiology of uncultured hydrothermal vent-associated methanotrophic bacteria. Using metagenomic binning techniques, we recovered and analyzed a near-complete genome from a novel gammaproteobacterial methanotroph (B42) associated with a white smoker chimney in the Southern Lau basin. B42 was the dominant methanotroph in the community, at ∼80x coverage, with only four others detected in the metagenome, all on low coverage contigs (7x–12x). Phylogenetic placement of B42 showed it is a member of the Methylothermaceae, a family currently represented by only one sequenced genome. Metabolic inferences based on the presence of known pathways in the genome showed that B42 possesses a branched respiratory chain with A- and B-family heme copper oxidases, cytochrome bd oxidase and a partial denitrification pathway. These genes could allow B42 to respire over a wide range of oxygen concentrations within the highly dynamic vent environment. Phylogenies of the denitrification genes revealed they are the result of separate horizontal gene transfer from other Proteobacteria and suggest that denitrification is a selective advantage in conditions where extremely low oxygen concentrations require all oxygen to be used for methane activation. PMID:26779119

  5. Global molecular analyses of methane metabolism in methanotrophic alphaproteobacterium, Methylosinus trichosporium OB3b. Part I: transcriptomic study

    Directory of Open Access Journals (Sweden)

    Janet B Matsen

    2013-04-01

    Full Text Available Methane-utilizing bacteria (methanotrophs are important in both environmental and biotechnological applications, due to their ability to convert methane to multicarbon compounds. However, systems-level studies of methane metabolism have not been carried out in methanotrophs. In this work we have integrated genomic and transcriptomic information to provide an overview of central metabolic pathways for methane utilization in Methylosinus trichosporium OB3b, a model alphaproteobacterial methanotroph. Particulate methane monooxygenase, PQQ-dependent methanol dehydrogenase, the H4MPT-pathway and NAD-dependent formate dehydrogenase are involved in methane oxidation to CO2. All genes essential for operation of the serine cycle, the ethylmalonyl-CoA (EMC pathway, and the citric acid (TCA cycle were expressed. PEP-pyruvate-oxaloacetate interconversions may have a function in regulation and balancing carbon between the serine cycle and the EMC pathway. A set of transaminases may contribute to carbon partitioning between the pathways. Metabolic pathways for acquisition and/or assimilation of nitrogen and iron are discussed.

  6. Methanotrophic bacteria associated to rice roots: the cultivar effect assessed by T-RFLP and microarray analysis.

    Science.gov (United States)

    Lüke, Claudia; Bodrossy, Levente; Lupotto, Elisabetta; Frenzel, Peter

    2011-10-01

    Rice plants play a key role in regulating methane emissions from paddy fields by affecting both underlying processes: methane production and oxidation. Specific differences were reported for methane oxidation rates; however, studies on the bacterial communities involved are rare. Here, we analysed the methanotrophic community on the roots of 18 different rice cultivars by pmoA-based terminal restriction fragment length polymorphism (T-RFLP) and microarray analysis. Both techniques showed comparable and consistent results revealing a high diversity dominated by type II and type Ib methanotrophs. pmoA microarrays have been successfully used to study methane-oxidizing bacteria in various environments. However, the microarray's full potential resolving community structure has not been exploited yet. Here, we provide an example on how to include this information into multivariate statistics. The analysis revealed a rice cultivar effect on the methanotroph community composition that could be affiliated to the plant genotype. This effect became only significant by including the specific phylogenetic resolution provided by the microarray into the statistical analysis.

  7. Ammonia Oxidation and Nitrite Reduction in the Verrucomicrobial Methanotroph Methylacidiphilum fumariolicum SolV

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    Sepehr S. Mohammadi

    2017-09-01

    Full Text Available The Solfatara volcano near Naples (Italy, the origin of the recently discovered verrucomicrobial methanotroph Methylacidiphilum fumariolicum SolV was shown to contain ammonium (NH4+ at concentrations ranging from 1 to 28 mM. Ammonia (NH3 can be converted to toxic hydroxylamine (NH2OH by the particulate methane monooxygenase (pMMO, the first enzyme of the methane (CH4 oxidation pathway. Methanotrophs rapidly detoxify the intermediate NH2OH. Here, we show that strain SolV performs ammonium oxidation to nitrite at a rate of 48.2 nmol NO2-.h−1.mg DW−1 under O2 limitation in a continuous culture grown on hydrogen (H2 as an electron donor. In addition, strain SolV carries out nitrite reduction at a rate of 74.4 nmol NO2-.h−1.mg DW−1 under anoxic condition at pH 5–6. This range of pH was selected to minimize the chemical conversion of nitrite (NO2- potentially occurring at more acidic pH values. Furthermore, at pH 6, we showed that the affinity constants (Ks of the cells for NH3 vary from 5 to 270 μM in the batch incubations with 0.5–8% (v/v CH4, respectively. Detailed kinetic analysis showed competitive substrate inhibition between CH4 and NH3. Using transcriptome analysis, we showed up-regulation of the gene encoding hydroxylamine dehydrogenase (haoA cells grown on H2/NH4+ compared to the cells grown on CH4/NO3- which do not have to cope with reactive N-compounds. The denitrifying genes nirk and norC showed high expression in H2/NH4+ and CH4/NO3- grown cells compared to cells growing at μmax (with no limitation while the norB gene showed downregulation in CH4/NO3- grown cells. These cells showed a strong upregulation of the genes in nitrate/nitrite assimilation. Our results demonstrate that strain SolV can perform ammonium oxidation producing nitrite. At high concentrations of ammonium this may results in toxic effects. However, at low oxygen concentrations strain SolV is able to reduce nitrite to N2O to cope with this toxicity.

  8. Transcriptional activities of methanogens and methanotrophs vary with methane emission flux in rice soils under chronic nutrient constraints of phosphorus and potassium

    Science.gov (United States)

    Sheng, Rong; Chen, Anlei; Zhang, Miaomiao; Whiteley, Andrew S.; Kumaresan, Deepak; Wei, Wenxue

    2016-12-01

    Nutrient status in soil is crucial for the growth and development of plants which indirectly or directly affect the ecophysiological functions of resident soil microorganisms. Soil methanogens and methanotrophs can be affected by soil nutrient availabilities and plant growth, which in turn modulate methane (CH4) emissions. Here, we assessed whether deficits in soil-available phosphorus (P) and potassium (K) modulated the activities of methanogens and methanotrophs in a long-term (20 year) experimental system involving limitation in either one or both nutrients. Results showed that a large amount of CH4 was emitted from paddy soil at rice tillering stage (flooding) while CH4 flux was minimum at ripening stage (drying). Compared to soils amended with NPK fertiliser treatment, the soils without P input significantly reduced methane flux rates, whereas those without K input did not. Under P limitation, methanotroph transcript copy number significantly increased in tandem with a decrease in methanogen transcript abundance, suggesting that P-deficiency-induced changes in soil physio-chemical properties, in tandem with rice plant growth, might constrain the activity of methanogens, whereas the methanotrophs might be adaptive to this soil environment. In contrast, lower transcript abundance of both methanogen and methanotrophs were observed in K-deficient soils. Assessments of community structures based upon transcripts indicated that soils deficient in P induced greater shifts in the active methanotrophic community than K-deficient soils, while similar community structures of active methanogens were observed in both treatments. These results suggested that the population dynamics of methanogens and methanotrophs could vary along with the changes in plant growth states and soil properties induced by nutrient deficiency.

  9. Fluorescence in situ hybridization-flow cytometry-cell sorting-based method for separation and enrichment of type I and type II methanotroph populations.

    Science.gov (United States)

    Kalyuzhnaya, Marina G; Zabinsky, Rebecca; Bowerman, Sarah; Baker, David R; Lidstrom, Mary E; Chistoserdova, Ludmila

    2006-06-01

    A fluorescence in situ hybridization-flow cytometry (FISH/FC)-based method was optimized using artificial mixtures of pure cultures of methanotrophic bacteria. Traditional oligonucleotide probes targeting 16S rRNAs of type I (MG84/705 probe) and type II (MA450 probe) methanotrophs were labeled with fluorescein or Alexa fluor and used for FISH, followed by fluorescence-activated FC analysis and cell sorting (FACS). The method resulted in efficient separation of target cells (type I or type II methanotrophs) from the artificial mixtures. The method was then applied for detection and enrichment of type I and type II methanotroph populations from a natural sample, Lake Washington sediment. Cells were extracted from the sediment, fixed, and subjected to FISH/FC/FACS. The resulting subpopulations were analyzed by reverse transcriptase PCR surveys of 16S rRNA, pmoA (encoding a subunit of particulate methane monooxygenase), and fae (encoding formaldehyde-activating enzyme) genes. The functional gene analysis indicated specific separation of the type I and type II methanotroph populations. 16S rRNA gene analysis revealed that type I methanotrophs comprised 59% of the subpopulation separated using the type I-specific probe and that type II methanotrophs comprised 47.5% of the subpopulation separated using the type II-specific probe. Our data indicate that the FISH/FC/FACS protocol described can provide significant enrichment of microbial populations of interest from complex natural communities and that these can be used for genetic tests. We further tested the possibility of direct whole-genome amplification (WGA) from limited numbers of sorted cells, using artificial mixtures of microbes whose genome sequences are known. We demonstrated that efficient WGA can be achieved using 10(4) or more cells separated by 16S rRNA-specific FISH/FC/FACS, while fewer cells resulted in less specific WGA.

  10. Growth rate characteristics of acidophilic heterotrophic organisms from mine waste rock piles

    Science.gov (United States)

    Yacob, T. W.; Silverstein, J.; Jenkins, J.; Andre, B. J.; Rajaram, H.

    2010-12-01

    Autotrophic iron oxidizing bacteria play a key role in pyrite oxidation and generation of acid mine drainage AMD. Scarcity of organic substrates in many disturbed sites insures that IOB have sufficient oxygen and other nutrients for growth. It is proposed that addition of organic carbon substrate to waste rock piles will result in enrichment of heterotrophic microorganisms limiting the role of IOB in AMD generation. Previous researchers have used the acidophilic heterotroph Acidiphilium cryptum as a model to study the effects of organic substrate addition on the pyrite oxidation/AMD cycle. In order to develop a quantitative model of effects such as competition for oxygen, it is necessary to use growth and substrate consumption rate expressions, and one approach is to choose a model strain such as A. cryptum for kinetic studies. However we have found that the growth rate characteristics of A. cryptum may not provide an accurate model of the remediation effects of organic addition to subsurface mined sites. Fluorescent in-situ hybridization (FISH) assays of extracts of mine waste rock enriched with glucose and yeast extract did not produce countable numbers of cells in the Acidiphilium genus, with a detection limit of3 x 104 cells/gram rock, despite evidence of the presence of well established heterotrophic organisms. However, an MPN enrichment produced heterotrophic population estimates of 1x107 and 1x109 cells/gram rock. Growth rate studies of A. cryptum showed that cultures took 120 hours to degrade 50% of an initial glucose concentration of 2,000 mg/L. However a mixed culture enriched from mine waste rock consumed 100% of the same amount of glucose in 24 hours. Substrate consumption data for the mixed culture were fit to a Monod growth model: {dS}/{dt} = μ_{max}S {( {X_0}/{Y} + S_0 -S )}/{(K_s +S)} Kinetic parameters were estimated utilizing a non linear regression method coupled with an ODE solver. The maximum specific growth rate of the mixed population with

  11. Quantifying adhesion of acidophilic bioleaching bacteria to silica and pyrite by atomic force microscopy with a bacterial probe.

    Science.gov (United States)

    Diao, Mengxue; Taran, Elena; Mahler, Stephen; Nguyen, Tuan A H; Nguyen, Anh V

    2014-03-01

    The adhesion of acidophilic bacteria to mineral surfaces is an important phenomenon in bioleaching processes. In this study, functionalized colloidal probes covered by bioleaching bacterial cells (Acidithiobacillus thiooxidans and Leptospirillum ferrooxidans) were developed and used to sense specific adhesion forces to a silica surface and a pyrite surface in various solutions. Experimentally, recorded retraction curves of A. thiooxidans revealed sawtooth features that were in good agreement with the wormlike chain model, while that of L. ferrooxidans exhibited stair-step separation. The magnitudes of adhesion forces and snap-off distances were strongly influenced by the ionic strength and pH. Macroscopic surface properties including hydrophobicity and surface potential for bacterial cells and substrata were measured by a sessile drop method and microelectrophoresis. The ATR-FTIR spectra indicated the presence of different types of biopolymers on two strains of bacteria.

  12. The Effect of Specific Conditions on Cu, Ni, Zn and Al Recovery from PCBS Waste Using Acidophilic Bacterial Strains

    Directory of Open Access Journals (Sweden)

    Mrážiková A.

    2016-03-01

    Full Text Available The objective of this work was to evaluate the influence of static, stirring and shaking conditions on copper, zinc, nickel and aluminium dissolution from printed circuit boards (PCBs using the mixed acidophilic bacterial culture of Acidithiobacillus ferrooxidans and Acidithiobacillus thiooxidans. The results revealed that static conditions were the most effective in zinc and aluminium dissolution. Zinc was removed almost completely under static conditions, whereas maximum of nickel dissolution was reached under the stirring conditions. The highest copper recovery (36% was reached under stirring conditions. The shaking conditions appeared to be the least suitable. The relative importance of these systems for the bioleaching of copper and nickel decreased in the order: stirring, static conditions, shaking.

  13. Abundance and Activity of Methanotrophic Bacteria in Littoral and Profundal Sediments of Lake Constance (Germany) ▿

    Science.gov (United States)

    Rahalkar, M.; Deutzmann, J.; Schink, B.; Bussmann, I.

    2009-01-01

    The abundances and activities of aerobic methane-oxidizing bacteria (MOB) were compared in depth profiles of littoral and profundal sediments of Lake Constance, Germany. Abundances were determined by quantitative PCR (qPCR) targeting the pmoA gene and by fluorescence in situ hybridization (FISH), and data were compared to methane oxidation rates calculated from high-resolution concentration profiles. qPCR using type I MOB-specific pmoA primers indicated that type I MOB represented a major proportion in both sediments at all depths. FISH indicated that in both sediments, type I MOB outnumbered type II MOB at least fourfold. Results obtained with both techniques indicated that in the littoral sediment, the highest numbers of methanotrophs were found at a depth of 2 to 3 cm, corresponding to the zone of highest methane oxidation activity, although no oxygen could be detected in this zone. In the profundal sediment, highest methane oxidation activities were found at a depth of 1 to 2 cm, while MOB abundance decreased gradually with sediment depth. In both sediments, MOB were also present at high numbers in deeper sediment layers where no methane oxidation activity could be observed. PMID:18997033

  14. Abundance and activity of methanotrophic bacteria in littoral and profundal sediments of lake constance (Germany).

    Science.gov (United States)

    Rahalkar, M; Deutzmann, J; Schink, B; Bussmann, I

    2009-01-01

    The abundances and activities of aerobic methane-oxidizing bacteria (MOB) were compared in depth profiles of littoral and profundal sediments of Lake Constance, Germany. Abundances were determined by quantitative PCR (qPCR) targeting the pmoA gene and by fluorescence in situ hybridization (FISH), and data were compared to methane oxidation rates calculated from high-resolution concentration profiles. qPCR using type I MOB-specific pmoA primers indicated that type I MOB represented a major proportion in both sediments at all depths. FISH indicated that in both sediments, type I MOB outnumbered type II MOB at least fourfold. Results obtained with both techniques indicated that in the littoral sediment, the highest numbers of methanotrophs were found at a depth of 2 to 3 cm, corresponding to the zone of highest methane oxidation activity, although no oxygen could be detected in this zone. In the profundal sediment, highest methane oxidation activities were found at a depth of 1 to 2 cm, while MOB abundance decreased gradually with sediment depth. In both sediments, MOB were also present at high numbers in deeper sediment layers where no methane oxidation activity could be observed.

  15. Characterization of the ectoine biosynthesis genes of haloalkalotolerant obligate methanotroph "Methylomicrobium alcaliphilum 20Z".

    Science.gov (United States)

    Reshetnikov, Alexander S; Khmelenina, Valentina N; Trotsenko, Yuri A

    2006-01-01

    The genes involved in biosynthesis of the major compatible solute ectoine (1,4,5,6-tetrahydro-2-methylpyrimidine carboxylic acid) in halotolerant obligate methanotroph "Methylomicrobium alcaliphilum 20Z" were studied. The complete nucleotide sequences of the structural genes encoding L: -aspartokinase (Ask), L-2,4-diaminobutyric acid transaminase (EctB), L-2,4-diaminobutyric acid acetyltransferase (EctA), and L-ectoine synthase (EctC) were defined and shown to be transcribed as a single operon ectABCask. Phylogenetic analysis revealed high sequence identities (34-63%) of the Ect proteins to those from halophilic heterotrophs with the highest amino acid identities being to Vibrio cholerae enzymes. The chromosomal DNA fragment from "M. alcaliphilum 20Z" containing ectABC genes and putative promoter region was expressed in Escherichia coli. Recombinant cells could grow in the presence of 4% NaCl and synthesize ectoine. The data obtained suggested that despite the ectoine biosynthesis pathway being evolutionary well conserved with respect to the genes and enzymes involved, some differences in their organization and regulation could occur in various halophilic bacteria.

  16. Two-liquid phase partitioning biotrickling filters for methane abatement: exploring the potential of hydrophobic methanotrophs.

    Science.gov (United States)

    Lebrero, Raquel; Hernández, Laura; Pérez, Rebeca; Estrada, José M; Muñoz, Raúl

    2015-03-15

    The potential of two-liquid phase biotrickling filters (BTFs) to overcome mass transfer limitations derived from the poor aqueous solubility of CH4 has been scarcely investigated to date. In this context, the abatement of diluted methane emissions in two-liquid phase BTFs was evaluated using two different inocula: a type II methanotrophs culture in BTF 1 and a hydrophobic microbial consortium capable of growing inside silicone oil in BTF 2. Both BTFs supported stable elimination capacities above 45 g m(-3) h(-1) regardless of the inoculum, whereas no improvement derived from the presence of hydrophobic microorganisms compared to the type II metanotrophs culture was observed. Interestingly, the addition of silicone oil mediated a reduced metabolites concentration in the recycling aqueous phase, thus decreasing the needs for mineral medium renewal. Moreover, a 78% similarity was recorded between the microbial communities enriched in both BTFs at the end of the experimental period in spite of the differences in the initial inoculum structure. The results obtained confirmed the superior performance of two-liquid phase BTFs for CH4 abatement compared with conventional biotrickling filters.

  17. Oxygen-dependent niche formation of a pyrite-dependent acidophilic consortium built by archaea and bacteria.

    Science.gov (United States)

    Ziegler, Sibylle; Dolch, Kerstin; Geiger, Katharina; Krause, Susanne; Asskamp, Maximilian; Eusterhues, Karin; Kriews, Michael; Wilhelms-Dick, Dorothee; Goettlicher, Joerg; Majzlan, Juraj; Gescher, Johannes

    2013-09-01

    Biofilms can provide a number of different ecological niches for microorganisms. Here, a multispecies biofilm was studied in which pyrite-oxidizing microbes are the primary producers. Its stability allowed not only detailed fluorescence in situ hybridization (FISH)-based characterization of the microbial population in different areas of the biofilm but also to integrate these results with oxygen and pH microsensor measurements conducted before. The O2 concentration declined rapidly from the outside to the inside of the biofilm. Hence, part of the population lives under microoxic or anoxic conditions. Leptospirillum ferrooxidans strains dominate the microbial population but are only located in the oxic periphery of the snottite structure. Interestingly, archaea were identified only in the anoxic parts of the biofilm. The archaeal community consists mainly of so far uncultured Thermoplasmatales as well as novel ARMAN (Archaeal Richmond Mine Acidophilic Nanoorganism) species. Inductively coupled plasma analysis and X-ray absorption near edge structure spectra provide further insight in the biofilm characteristics but revealed no other major factors than oxygen affecting the distribution of bacteria and archaea. In addition to catalyzed reporter deposition FISH and oxygen microsensor measurements, microautoradiographic FISH was used to identify areas in which active CO2 fixation takes place. Leptospirilla as well as acidithiobacilli were identified as primary producers. Fixation of gaseous CO2 seems to proceed only in the outer rim of the snottite. Archaea inhabiting the snottite core do not seem to contribute to the primary production. This work gives insight in the ecological niches of acidophilic microorganisms and their role in a consortium. The data provided the basis for the enrichment of uncultured archaea.

  18. Changes in methane oxidation activity and methanotrophic community composition in saline alkaline soils.

    Science.gov (United States)

    Serrano-Silva, Nancy; Valenzuela-Encinas, César; Marsch, Rodolfo; Dendooven, Luc; Alcántara-Hernández, Rocio J

    2014-05-01

    The soil of the former Lake Texcoco is a saline alkaline environment where anthropogenic drainage in some areas has reduced salt content and pH. Potential methane (CH4) consumption rates were measured in three soils of the former Lake Texcoco with different electrolytic conductivity (EC) and pH, i.e. Tex-S1 a >18 years drained soil (EC 0.7 dS m(-1), pH 8.5), Tex-S2 drained for ~10 years (EC 9.0 dS m(-1), pH 10.3) and the undrained Tex-S3 (EC 84.8 dS m(-1), pH 10.3). An arable soil from Alcholoya (EC 0.7 dS m(-1), pH 6.7), located nearby Lake Texcoco was used as control. Methane oxidation in the soil Tex-S1 (lowest EC and pH) was similar to that in the arable soil from Alcholoya (32.5 and 34.7 mg CH4 kg(-1) dry soil day(-1), respectively). Meanwhile, in soils Tex-S2 and Tex-S3, the potential CH4 oxidation rates were only 15.0 and 12.8 mg CH4 kg(-1) dry soil day(-1), respectively. Differences in CH4 oxidation were also related to changes in the methane-oxidizing communities in these soils. Sequence analysis of pmoA gene showed that soils differed in the identity and number of methanotrophic phylotypes. The Alcholoya soil and Tex-S1 contained phylotypes grouped within the upland soil cluster gamma and the Jasper Ridge, California JR-2 clade. In soil Tex-S3, a phylotype related to Methylomicrobium alcaliphilum was detected.

  19. High resolution depth distribution of Bacteria, Archaea, methanotrophs, and methanogens in the bulk and rhizosphere soils of a flooded rice paddy

    Directory of Open Access Journals (Sweden)

    Hyo Jung eLee

    2015-06-01

    Full Text Available The communities and abundances of methanotrophs and methanogens, along with the oxygen, methane, and total organic carbon (TOC concentrations, were investigated along a depth gradient in a flooded rice paddy. Broad patterns in vertical profiles of oxygen, methane, TOC, and microbial abundances were similar in the bulk and rhizosphere soils, though methane and TOC concentrations and 16S rRNA gene copies were clearly higher in the rhizosphere soil than in the bulk soil. Oxygen concentrations decreased sharply to below detection limits at the 8 mm depth. Pyrosequencing of 16S rRNA genes showed that bacterial and archaeal communities varied according to the oxic, oxic-anoxic, and anoxic zones, indicating that oxygen is a determining factor for the distribution of bacterial and archaeal communities. Aerobic methanotrophs were maximally observed near the oxic-anoxic interface, while methane, TOC, and methanogens were highest in the rhizosphere soil at 30–200 mm depth, suggesting that methane is produced mainly from organic carbon derived from rice plants and is metabolized aerobically. The relative abundances of type I methanotrophs such as Methylococcus, Methylomonas, and Methylocaldum decreased more drastically than those of type II methanotrophs (such as Methylocystis and Methylosinus with increasing depth. Methanosaeta and Methanoregula were predominant methanogens at all depths, and the relative abundances of Methanosaeta, Methanoregula, and Methanosphaerula, and GOM_Arc_I increased with increasing depth. Based on contrasts between absolute abundances of methanogens and methanotrophs at depths sampled across rhizosphere and bulk soils (especially millimeter-scale slices at the surface, we have identified populations of methanogens (Methanosaeta, Methanoregula, Methanocella, Methanobacterium, and Methanosphaerula and methanotrophs (Methylosarcina, Methylococcus, Methylosinus, and unclassified Methylocystaceae that are likely physiologically

  20. Production of Glycolic Acid by Chemolithotrophic Iron- and Sulfur-Oxidizing Bacteria and Its Role in Delineating and Sustaining Acidophilic Sulfide Mineral-Oxidizing Consortia▿

    Science.gov (United States)

    Ñancucheo, Ivan; Johnson, D. Barrie

    2010-01-01

    Glycolic acid was detected as an exudate in actively growing cultures of three chemolithotrophic acidophiles that are important in biomining operations, Leptospirillum ferriphilum, Acidithiobacillus (At.) ferrooxidans, and At. caldus. Although similar concentrations of glycolic acid were found in all cases, the concentrations corresponded to ca. 24% of the total dissolved organic carbon (DOC) in cultures of L. ferriphilum but only ca. 5% of the total DOC in cultures of the two Acidithiobacillus spp. Rapid acidification (to pH 1.0) of the culture medium of At. caldus resulted in a large increase in the level of DOC, although the concentration of glycolic acid did not change in proportion. The archaeon Ferroplasma acidiphilum grew in the cell-free spent medium of At. caldus; glycolic acid was not metabolized, although other unidentified compounds in the DOC pool were metabolized. Glycolic acid exhibited levels of toxicity with 21 strains of acidophiles screened similar to those of acetic acid. The most sensitive species were chemolithotrophs (L. ferriphilum and At. ferrivorans), while the most tolerant species were chemoorganotrophs (Acidocella, Acidobacterium, and Ferroplasma species), and the ability to metabolize glycolic acid appeared to be restricted (among acidophiles) to Firmicutes (chiefly Sulfobacillus spp.). Results of this study help explain why Sulfobacillus spp. rather than other acidophiles are the main organic carbon-degrading bacteria in continuously fed stirred tanks used to bioprocess sulfide mineral concentrates and also why temporary cessation of pH control in these systems, resulting in rapid acidification, often results in a plume of the archaeon Ferroplasma. PMID:19933342

  1. Production of glycolic acid by chemolithotrophic iron- and sulfur-oxidizing bacteria and its role in delineating and sustaining acidophilic sulfide mineral-oxidizing consortia.

    Science.gov (United States)

    Nancucheo, Ivan; Johnson, D Barrie

    2010-01-01

    Glycolic acid was detected as an exudate in actively growing cultures of three chemolithotrophic acidophiles that are important in biomining operations, Leptospirillum ferriphilum, Acidithiobacillus (At.) ferrooxidans, and At. caldus. Although similar concentrations of glycolic acid were found in all cases, the concentrations corresponded to ca. 24% of the total dissolved organic carbon (DOC) in cultures of L. ferriphilum but only ca. 5% of the total DOC in cultures of the two Acidithiobacillus spp. Rapid acidification (to pH 1.0) of the culture medium of At. caldus resulted in a large increase in the level of DOC, although the concentration of glycolic acid did not change in proportion. The archaeon Ferroplasma acidiphilum grew in the cell-free spent medium of At. caldus; glycolic acid was not metabolized, although other unidentified compounds in the DOC pool were metabolized. Glycolic acid exhibited levels of toxicity with 21 strains of acidophiles screened similar to those of acetic acid. The most sensitive species were chemolithotrophs (L. ferriphilum and At. ferrivorans), while the most tolerant species were chemoorganotrophs (Acidocella, Acidobacterium, and Ferroplasma species), and the ability to metabolize glycolic acid appeared to be restricted (among acidophiles) to Firmicutes (chiefly Sulfobacillus spp.). Results of this study help explain why Sulfobacillus spp. rather than other acidophiles are the main organic carbon-degrading bacteria in continuously fed stirred tanks used to bioprocess sulfide mineral concentrates and also why temporary cessation of pH control in these systems, resulting in rapid acidification, often results in a plume of the archaeon Ferroplasma.

  2. Activities of methionine-γ-lyase in the acidophilic archaeon “Ferroplasma acidarmanus” strain fer1

    Directory of Open Access Journals (Sweden)

    Khan MA

    2013-04-01

    Full Text Available M A Khan,1 Madeline M López-Muñoz,2 Charles W Kaspar,3 Kai F Hung1 1Department of Biological Sciences, Eastern Illinois University, Charleston, IL, USA; 2Department of Biology, Universidad de Puerto Rico, Mayaguez, Puerto Rico; 3Bacteriology Department, University of Wisconsin, Madison, WI, USA Abstract: Biogeochemical processes on exposed pyrite ores result in extremely high levels of sulfuric acid at these locations. Acidophiles that thrive in these conditions must overcome significant challenges, including an environment with proton concentrations at pH 3 or below. The role of sulfur metabolism in the archaeon “Ferroplasma acidarmanus” strain fer1's ability to thrive in this environment was investigated due to its growth-dependent production of methanethiol, a volatile organic sulfur compound. Two putative sequences for methionine-γ-lyase (EC 4.4.1.11, an enzyme known to carry out α, γ-elimination on L-methionine to produce methanethiol, were identified in fer1. Bioinformatic analyses identified a conserved pyridoxal-5'-phosphate (PLP binding domain and a partially conserved catalytic domain in both putative sequences. Detection of PLP-dependent and L-methionine-dependent production of α-keto compounds and thiol groups in fer1 confirmed the presence of methionine-γ-lyase activity. Further, fer1 lysate was capable of processing related substrates, including D-methionine, L-cysteine, L-cystathionine, and L/D-homocysteine. When the two putative fer1 methionine-γ-lyase gene-coded proteins were expressed in Escherichia coli cells, one sequence demonstrated an ability to carry out α, γ-elimination activity, while the other exhibited γ-replacement activity. These fer1 methionine-γ-lyases also exhibited optimum pH, substrate specificity, and catalytic preferences that are different from methionine-γ-lyases from other organisms. These differences are discussed in the context of molecular phylogeny constructed using a maximum

  3. Use of an acidophilic yeast strain to enable the growth of leaching bacteria on solid media.

    Science.gov (United States)

    Ngom, Baba; Liang, Yili; Liu, Yi; Yin, Huaqun; Liu, Xueduan

    2015-03-01

    In this study, a Candida digboiensis strain was isolated from a heap leaching plant in Zambia and used in double-layer agar plate to efficiently isolate and purify leaching bacteria. Unlike Acidiphilium sp., the yeast strain was tetrathionate tolerant and could metabolize a great range of organic compounds including organic acids. These properties allowed the yeast strain to enable and fasten the growth of iron and sulfur oxidizers on double-layer agar plate. The isolates were identified as Acidithiobacillus ferrooxidans FOX1, Leptospirillun ferriphilum BN, and Acidithiobacillus thiooxidans ZMB. These three leaching bacteria were inhibited by organic acids such as acetic and propionic acids; however, their activities were enhanced by Candida digboiensis NB under dissolved organic matter stress.

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

    KAUST Repository

    Meulepas, Roel J W

    2010-05-06

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

  5. Enhanced Productivity of a Lutein-Enriched Novel Acidophile Microalga Grown on Urea

    NARCIS (Netherlands)

    Casal, C.; Cuaresma, M.; Vega, J.M.; Vilchez, C.

    2011-01-01

    Coccomyxa acidophila is an extremophile eukaryotic microalga isolated from the Tinto River mining area in Huelva, Spain. Coccomyxa acidophila accumulates relevant amounts of b-carotene and lutein, well-known carotenoids with many biotechnological applications, especially in food and health-related

  6. Enhanced Productivity of a Lutein-Enriched Novel Acidophile Microalga Grown on Urea

    NARCIS (Netherlands)

    Casal, C.; Cuaresma, M.; Vega, J.M.; Vilchez, C.

    2011-01-01

    Coccomyxa acidophila is an extremophile eukaryotic microalga isolated from the Tinto River mining area in Huelva, Spain. Coccomyxa acidophila accumulates relevant amounts of b-carotene and lutein, well-known carotenoids with many biotechnological applications, especially in food and health-related i

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

    Energy Technology Data Exchange (ETDEWEB)

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

    2010-07-01

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

  8. Metabolites Associated with Adaptation of Microorganisms to an Acidophilic, Metal-Rich Environment Identified by Stable-Isotope-Enabled Metabolomics

    Science.gov (United States)

    Mosier, Annika C.; Justice, Nicholas B.; Bowen, Benjamin P.; Baran, Richard; Thomas, Brian C.; Northen, Trent R.; Banfield, Jillian F.

    2013-01-01

    ABSTRACT Microorganisms grow under a remarkable range of extreme conditions. Environmental transcriptomic and proteomic studies have highlighted metabolic pathways active in extremophilic communities. However, metabolites directly linked to their physiology are less well defined because metabolomics methods lag behind other omics technologies due to a wide range of experimental complexities often associated with the environmental matrix. We identified key metabolites associated with acidophilic and metal-tolerant microorganisms using stable isotope labeling coupled with untargeted, high-resolution mass spectrometry. We observed >3,500 metabolic features in biofilms growing in pH ~0.9 acid mine drainage solutions containing millimolar concentrations of iron, sulfate, zinc, copper, and arsenic. Stable isotope labeling improved chemical formula prediction by >50% for larger metabolites (>250 atomic mass units), many of which were unrepresented in metabolic databases and may represent novel compounds. Taurine and hydroxyectoine were identified and likely provide protection from osmotic stress in the biofilms. Community genomic, transcriptomic, and proteomic data implicate fungi in taurine metabolism. Leptospirillum group II bacteria decrease production of ectoine and hydroxyectoine as biofilms mature, suggesting that biofilm structure provides some resistance to high metal and proton concentrations. The combination of taurine, ectoine, and hydroxyectoine may also constitute a sulfur, nitrogen, and carbon currency in the communities. PMID:23481603

  9. Organization and regulation of the arsenite oxidase operon of the moderately acidophilic and facultative chemoautotrophic Thiomonas arsenitoxydans.

    Science.gov (United States)

    Slyemi, Djamila; Moinier, Danielle; Talla, Emmanuel; Bonnefoy, Violaine

    2013-11-01

    Thiomonas arsenitoxydans is an acidophilic and facultatively autotrophic bacterium that can grow by oxidizing arsenite to arsenate. A comparative genomic analysis showed that the T. arsenitoxydans aioBA cluster encoding the two subunits of arsenite oxidase is distinct from the other clusters, with two specific genes encoding a cytochrome c and a metalloregulator belonging to the ArsR/SmtB family. These genes are cotranscribed with aioBA, suggesting that these cytochromes c are involved in arsenite oxidation and that this operon is controlled by the metalloregulator. The growth of T. arsenitoxydans in the presence of thiosulfate and arsenite, or arsenate, is biphasic. Real-time PCR experiments showed that the operon is transcribed during the second growth phase in the presence of arsenite or arsenate, whereas antimonite had no effect. These results suggest that the expression of the aioBA operon of T. arsenitoxydans is regulated by the electron donor present in the medium, i.e., is induced in the presence of arsenic but is repressed by more energetic substrates. Our data indicate that the genetic organization and regulation of the aioBA operon of T. arsenitoxydans differ from those of the other arsenite oxidizers.

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

    Directory of Open Access Journals (Sweden)

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

    2010-07-01

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

  11. Effects of nitrogen application rate and a nitrification inhibitor dicyandiamide on methanotroph abundance and methane uptake in a grazed pasture soil.

    Science.gov (United States)

    Dai, Yu; Di, Hong J; Cameron, Keith C; He, Ji-Zheng

    2013-12-01

    Methane-oxidizing bacteria (methanotrophs) in the soil are a unique group of methylotrophic bacteria that utilize methane (CH4) as their sole source of carbon and energy which limit the flux of methane to the atmosphere from soils and consume atmospheric methane. A field experiment was conducted to determine the effect of nitrogen application rates and the nitrification inhibitor dicyandiamide (DCD) on the abundance of methanotrophs and on methane flux in a grazed pasture soil. Nitrogen (N) was applied at four different rates, with urea applied at 50 and 100 kg N ha(-1) and animal urine at 300 and 600 kg N ha(-1). DCD was applied at 10 kg ha(-1). The results showed that both the DNA and selected mRNA copy numbers of the methanotroph pmoA gene were not affected by the application of urea, urine or DCD. The methanotroph DNA and mRNA pmoA gene copy numbers were low in this soil, below 7.13 × 10(3) g(-1) soil and 3.75 × 10(3) μg(-1) RNA, respectively. Daily CH4 flux varied slightly among different treatments during the experimental period, ranging from -12.89 g CH4 ha(-1) day(-1) to -0.83 g CH4 ha(-1) day(-1), but no significant treatment effect was found. This study suggests that the application of urea fertilizer, animal urine returns and the use of the nitrification inhibitor DCD do not significantly affect soil methanotroph abundance or daily CH4 fluxes in grazed grassland soils.

  12. Unmodified prolactin (PRL) promotes PRL secretion and acidophil hypertrophy and is associated with pituitary hyperplasia in female rats.

    Science.gov (United States)

    Johnson, Terence E; Vue, Mayza; Brekhus, Sharyn; Khong, Amy; Ho, Timothy W C; Walker, Ameae M

    2003-01-01

    In this study, we have tested the hypothesis that unmodified prolactin (U-PRL) and phosphorylated prolactin (P-PRL) have differential roles in the autoregulation of PRL secretion in vivo. Recombinant human U-PRL and a molecular mimic of P-PRL (S179D PRL) were administered to male rats and to female rats in different physiological states and the effect on rat PRL release was measured. Administration of U-PRL elevated rat PRL in all female animals, but was without effect in males. By contrast, S179D PRL was inactive in females, but inhibited PRL release in males. Morphometric and immunohistochemical analyses demonstrated acidophil hypertrophy and evidence of increased PRL secretion in the pituitaries of U-PRL-treated females. Analysis of the two forms of PRL during prolactinoma induction in two differentially susceptible strains of rats found a strong temporal correlation among increased ratios of U-PRL: P-PRL, increased circulating PRL, and increased cell proliferation. We conclude (1). that the autoregulatory mechanism(s) can distinguish between the two major forms of PRL and that higher proportions of U-PRL not only allow for higher circulating levels of PRL, but are also autostimulatory, (2). that the autoregulatory mechanism( s) are set differently in males and females such that females are more sensitive to autostimulation by U-PRL and less sensitive to inhibition by P-PRL, and (3). that U-PRL and P-PRL may also have differential roles in the regulation of pituitary cell proliferation.

  13. Proteogenomic and functional analysis of chromate reduction in Acidiphilium cryptum JF-5, an Fe(III)-respiring acidophile.

    Science.gov (United States)

    Magnuson, Timothy S; Swenson, Michael W; Paszczynski, Andrzej J; Deobald, Lee A; Kerk, David; Cummings, David E

    2010-12-01

    Acidiphilium cryptum JF-5, an acidophilic iron-respiring Alphaproteobacterium, has the ability to reduce chromate under aerobic and anaerobic conditions, making it an intriguing and useful model organism for the study of extremophilic bacteria in bioremediation applications. Genome sequence annotation suggested two potential mechanisms of Cr(VI) reduction, namely, a number of c-type cytochromes, and a predicted NADPH-dependent Cr(VI) reductase. In laboratory studies using pure cultures of JF-5, an NADPH-dependent chromate reductase activity was detected primarily in soluble protein fractions, and a periplasmic c-type cytochrome (ApcA) was also present, representing two potential means of Cr(VI) reduction. Upon further examination, it was determined that the NADPH-dependent activity was not specific for Cr(VI), and the predicted proteins were not detected in Cr(VI)-grown cultures. Proteomic data did show measureable amounts of ApcA in cells grown with Cr(VI). Purified ApcA is reducible by menadiol, and in turn can reduce Cr(VI), suggesting a means to obtain electrons from the respiratory chain and divert them to Cr(VI). Electrochemical measurements confirm that Cr reduction by ApcA is pH dependent, with low pH being favored. Homology modeling of ApcA and comparison to a known Cr(VI)-reducing c-type cytochrome structure revealed basic amino acids which could interact with chromate ion. From these studies, it can be concluded that A. cryptum has the physiologic and genomic capability to reduce Cr(VI) to the less toxic Cr(III). However, the expected chromate reductase mechanism may not be the primary means of Cr(VI) reduction in this organism.

  14. Genomic and physiological analysis of carbon storage in the verrucomicrobial methanotroph "Ca. Methylacidiphilum fumariolicum" SolV

    Directory of Open Access Journals (Sweden)

    Ahmad Fouad eKhadem

    2012-09-01

    Full Text Available Candidatus Methylacidiphilum fumariolicum SolV is a verrucomicrobial methanotroph that can grow in extremely acidic environments at high temperature. Strain SolV fixes carbon dioxide (CO2 via the Calvin-Benson-Bassham cycle with methane as energy source, a trait so far very unusual in methanotrophs. In this study, the ability of Ca. M. fumariolicum to store carbon was explored by genome analysis, physiological studies and electron microscopy. When cell cultures were depleted for nitrogen, glycogen storage was clearly observed in cytoplasmic storage vesicles by electron microscopy. After cessation of growth, the dry weight kept increasing and the bacteria were filled up almost entirely by glycogen. This was confirmed by biochemical analysis, which showed that glycogen accumulated to 36% of the total dry weight of the cells. When methane was removed from the culture, this glycogen was consumed within 47 days. During the period of glycogen consumption, the bacteria kept their viability high when compared to bacteria without glycogen (from cultures growing exponentially. The latter bacteria lost viability already after a few days when starved for methane. Analysis of the draft genome of Ca. M. fumariolicum SolV demonstrated that all known genes for glycogen storage and degradation were present and also transcribed. Phylogenetic analysis of these genes showed that they form a separate cluster with Ca. M. infernorum V4, and the most closely related other sequences only have an identity of 40%. This study presents the first physiological evidence of glycogen storage in the phylum Verrucomicrobia and indicates that carbon storage is important for survival at times of methane starvation.

  15. [Effect of temperature on the rate of oxidation of pyrrhotite-rich sulfide ore flotation concentrate and the structure of the acidophilic chemolithoautotrophic microbial community].

    Science.gov (United States)

    Moshchanetskii, P V; Pivovarova, T A; Belyi, A V; Kondrat'eva, T F

    2014-01-01

    Oxidation of flotation concentrate of a pyrrhotite-rich sulfide ore by acidophilic chemolithoautotrophic microbial communities at 35, 40, and 45 degrees C was investigated. According to the physicochemical parameters of the liquid phase of the pulp, as well as the results of analysis of the solid residue after biooxidation and cyanidation, the community developed at 40 degrees C exhibited the highest rate of oxidation. The degree of gold recovery at 35, 40, and 45 degrees C was 89.34, 94.59, and 83.25%, respectively. At 40 degrees C, the highest number of microbial cells (6.01 x 10(9) cells/mL) was observed. While temperature had very little effect on the species composition of microbial communities, except for the absence of Leptospirillum ferriphilum at 35 degrees C, the shares of individual species in the communities varied with temperature. Relatively high numbers of Sulfobacillus thermosulfidooxidans, the organism oxidizing iron and elemental sulfur at higher rates than other acidophilic chemolithotrophic species, were observed at 40 degrees C.

  16. GLOBAL MOLECULAR ANALYSES OF METHANE METABOLISM IN METHANOTROPHIC ALPHAPROTEOBACTERIUM, METHYLOSINUS TRICHOSPORIUM OB3B.PART II. METABOLOMICS AND 13C-LABELING STUDY

    Directory of Open Access Journals (Sweden)

    Marina G. Kalyuzhanaya

    2013-04-01

    Full Text Available In this work we use metabolomics and 13C-labeling data to refine central metabolic pathways for methane utilization in Methylosinus trichosporium OB3b, a model alphaproteobacterial methanotrophic bacterium. We demonstrate here that similar to non-methane utilizing methylotrophic alphaproteobacteria the core metabolism of the microbe is represented by several tightly connected metabolic cycles, such as the serine pathway, the ethylmalonyl-CoA (EMC pathway, and the citric acid (TCA cycle. Both in silico estimations and stable isotope labeling experiments combined with single cell (NanoSIMS and bulk biomass analyses indicate that a significantly larger portion of the cell carbon (over 60% is derived from CO2 in this methanotroph. Our 13C-labeling studies revealed an unusual topology of the assimilatory network in which phosph(enolpyruvate/pyruvate interconversions are key metabolic switches. A set of additional pathways for carbon fixation are identified and discussed.

  17. Cultivation of acidophilic algae Galdieria sulphuraria and Pseudochlorella sp. YKT1 in media derived from acidic hot springs

    Directory of Open Access Journals (Sweden)

    Shunsuke Hirooka

    2016-12-01

    Full Text Available Microalgae possess a high potential for producing pigments, antioxidants, and lipophilic compounds for industrial applications. However, the cultivation of microalgae comes at a high cost. To reduce the cost, changes from a closed bioreactor to open pond system and from a synthetic medium to environmental or wastewater-based medium are being sought. However, the use of open pond systems is currently limited because of contamination by undesirable organisms. To overcome this issue, one strategy is to combine acidophilic algae and acidic drainage in which other organisms are unable to thrive. Here, we tested waters from sulfuric acidic hot springs (Tamagawa, pH 1.15 and Tsukahara, pH 1.14 in Japan for the cultivation of the red alga Galdieria sulphuraria 074G and the green alga Pseudochlorella sp. YKT1. Both of these spring waters are rich in phosphate (0.043 and 0.145 mM, respectively compared to other environmental freshwater sources. Neither alga grew in the spring water but they grew very well when the waters were supplemented with an inorganic nitrogen source. The algal yields were ~2.73 g dry weight/L for G. sulphuraria and ~2.49 g dry weight/L for P. sp. YKT1, which were comparable to those in an autotrophic synthetic medium. P. sp. YKT1 grew in the spring waters supplemented either of NH4+, NO3- or urea, while G. sulphuraria grew only when NH4+ was supplemented. For P. sp. YKT1, the spring water was adjusted to pH 2.0, while for G. sulphuraria, no pH adjustment was required. In both cases, no additional pH-buffering compound was required. The phycocyanin of the thermophilic G. sulphuraria is known to be more thermostable than that from the Spirulina platensis currently used in phycocyanin production for commercial use. The phycocyanin content in G. sulphuraria in the Tsukahara water supplemented with NH4+ was 107.42±1.81 μg/mg dry weight, which is comparable to the level in S. platensis (148.3 μg/mg dry weight. P. sp. YKT1 cells in the

  18. Cultivation of Acidophilic Algae Galdieria sulphuraria and Pseudochlorella sp. YKT1 in Media Derived from Acidic Hot Springs.

    Science.gov (United States)

    Hirooka, Shunsuke; Miyagishima, Shin-Ya

    2016-01-01

    Microalgae possess a high potential for producing pigments, antioxidants, and lipophilic compounds for industrial applications. However, the cultivation of microalgae comes at a high cost. To reduce the cost, changes from a closed bioreactor to open pond system and from a synthetic medium to environmental or wastewater-based medium are being sought. However, the use of open pond systems is currently limited because of contamination by undesirable organisms. To overcome this issue, one strategy is to combine acidophilic algae and acidic drainage in which other organisms are unable to thrive. Here, we tested waters from sulfuric acidic hot springs (Tamagawa, pH 1.15 and Tsukahara, pH 1.14) in Japan for the cultivation of the red alga Galdieria sulphuraria 074G and the green alga Pseudochlorella sp. YKT1. Both of these spring waters are rich in phosphate (0.043 and 0.145 mM, respectively) compared to other environmental freshwater sources. Neither alga grew in the spring water but they grew very well when the waters were supplemented with an inorganic nitrogen source. The algal yields were ∼2.73 g dry weight/L for G. sulphuraria and ∼2.49 g dry weight/L for P. sp. YKT1, which were comparable to those in an autotrophic synthetic medium. P. sp. YKT1 grew in the spring waters supplemented either of NH4(+), NO3(-) or urea, while G. sulphuraria grew only when NH4(+) was supplemented. For P. sp. YKT1, the spring water was adjusted to pH 2.0, while for G. sulphuraria, no pH adjustment was required. In both cases, no additional pH-buffering compound was required. The phycocyanin of the thermophilic G. sulphuraria is known to be more thermostable than that from the Spirulina platensis currently used in phycocyanin production for commercial use. The phycocyanin content in G. sulphuraria in the Tsukahara water supplemented with NH4(+) was 107.42 ± 1.81 μg/mg dry weight, which is comparable to the level in S. platensis (148.3 μg/mg dry weight). P. sp. YKT1 cells in the

  19. Genome sequence of the copper resistant and acid-tolerant Desulfosporosinus sp. BG isolated from the tailings of a molybdenum-tungsten mine in the Transbaikal area

    Directory of Open Access Journals (Sweden)

    Olga V. Karnachuk

    2017-03-01

    Full Text Available Here, we report on the draft genome of a copper-resistant and acidophilic Desulfosporosinus sp. BG, isolated from the tailings of a molybdenum-tungsten mine in Transbaikal area. The draft genome has a size of 4.52 Mb and encodes transporters of heavy metals. The phylogenetic analysis based on concatenated ribosomal proteins revealed that strain BG clusters together with the other acidophilic copper-resistant strains Desulfosporosinus sp. OT and Desulfosporosinus sp. I2. The K+-ATPase, Na+/H+ antiporter and amino acid decarboxylases may participate in enabling growth at low pH. The draft genome sequence and annotation have been deposited at GenBank under the accession number NZ_MASS00000000.

  20. Methanotrophic activity in the water column above shallow gas flares west of Prins Karls Forland, Arctic Ocean

    Science.gov (United States)

    Gründger, Friederike; Svenning, Mette M.; Niemann, Helge; Silyakova, Anna; Serov, Pavel; Pavlov, Alexey K.; Granskog, Mats A.; Ferre, Bénédicte; Carroll, JoLynn

    2016-04-01

    Numerous gas flares, interpreted to be streams of methane bubbles, were discovered in shallow waters (average water depth about 90 m) on the continental shelf west of Prins Karls Forland (Western Svalbard) in the Arctic Ocean. Gas is released from the seabed to the water column and potentially transferred into the atmosphere where it acts as a potent greenhouse gas. In order to resolve the fate of dissolved methane in the water column, we carried out grid-pattern biogeochemical measurements in the study area of 30 x 15 km. Specifically, we measured concentrations of dissolved methane and microbial methane oxidation (MOx) rates at 8 water depths at 31 sampling stations and performed 16S rRNA sequencing analysis on selected samples to characterize the microbial community composition. Availability of dissolved methane is essential for the process of microbial methane oxidation. However, our measurements reveal that high concentrations of dissolved methane in the water column do not necessarily lead to high MOx rates. Our results indicated that the presence of marine methanotrophic biomass as well as dissolved organic matter is of larger importance for the process of microbial methane oxidation. For example, we found MOx hot spots with values up to 13 nmol l-1 d-1 at bottom water depth with dissolved methane concentrations less than 160 nmol l-1. In contrast, at stations where bottom methane concentration values reached 640 nmol l-1, MOx rates were less than 0.7 nmol l-1 d-1. To interpret observed interconnection between methane concentrations and MOx rates, we use vertical distributions of seawater temperature, salinity and properties of colored dissolved organic matter (CDOM). This information helps us characterize the oceanographic setting and circulation patterns in the area, which we believe has a major impact on the origin and distribution of methanotrophic microbial biomass and methane oxidation in methanerich bottom water. This study is part of the Centre for

  1. Methane-Fueled Syntrophy through Extracellular Electron Transfer: Uncovering the Genomic Traits Conserved within Diverse Bacterial Partners of Anaerobic Methanotrophic Archaea.

    Science.gov (United States)

    Skennerton, Connor T; Chourey, Karuna; Iyer, Ramsunder; Hettich, Robert L; Tyson, Gene W; Orphan, Victoria J

    2017-08-01

    The anaerobic oxidation of methane by anaerobic methanotrophic (ANME) archaea in syntrophic partnership with deltaproteobacterial sulfate-reducing bacteria (SRB) is the primary mechanism for methane removal in ocean sediments. The mechanism of their syntrophy has been the subject of much research as traditional intermediate compounds, such as hydrogen and formate, failed to decouple the partners. Recent findings have indicated the potential for extracellular electron transfer from ANME archaea to SRB, though it is unclear how extracellular electrons are integrated into the metabolism of the SRB partner. We used metagenomics to reconstruct eight genomes from the globally distributed SEEP-SRB1 clade of ANME partner bacteria to determine what genomic features are required for syntrophy. The SEEP-SRB1 genomes contain large multiheme cytochromes that were not found in previously described free-living SRB and also lack periplasmic hydrogenases that may prevent an independent lifestyle without an extracellular source of electrons from ANME archaea. Metaproteomics revealed the expression of these cytochromes at in situ methane seep sediments from three sites along the Pacific coast of the United States. Phylogenetic analysis showed that these cytochromes appear to have been horizontally transferred from metal-respiring members of the Deltaproteobacteria such as Geobacter and may allow these syntrophic SRB to accept extracellular electrons in place of other chemical/organic electron donors.IMPORTANCE Some archaea, known as anaerobic methanotrophs, are capable of converting methane into carbon dioxide when they are growing syntopically with sulfate-reducing bacteria. This partnership is the primary mechanism for methane removal in ocean sediments; however, there is still much to learn about how this syntrophy works. Previous studies have failed to identify the metabolic intermediate, such as hydrogen or formate, that is passed between partners. However, recent analysis of

  2. Characterization of bacteria acidophilic in samples of water coming into a region that suffers influence of uranium mine in Caldas (MG)

    Energy Technology Data Exchange (ETDEWEB)

    Campos, Michelle B.; Ferrari, Carla R.; Roque, Claudio V.; Ronqui, Leilane B.; Nascimento, Marcos R.L. do; Rodgher, Suzelei; Azevedo, Heliana [Laboratorio de Pocos de Caldas (LAPOC-CNEN/MG), MG (Brazil)], e-mail: michelle_borato@hotmail.com, e-mail: carlarolimferrari@yahoo.com.br, e-mail: cvroque@cnen.gov.br, e-mail: leilanebio@yahoo.com.br, e-mail: pmarcos@cnen.gov.br, e-mail: surodgher@uol.com.br, e-mail: hgomes@cnen.gov.br

    2009-07-01

    The fundamental condition for the bioleaching of the uranium ore is the presence of metallic sulfide such as pyrite associated with the ore, which is found in the ore and in the waste at the Unidade de Tratamento de Minerio (UTM) of Pocos de Caldas, State of Minas Gerais, Brazil. The present study aims to determine the chemical and microbiological characteristics in effluents of uranium mining from the UTM and in Antas dam, which receives treated effluents from the UTM. Water samples were collected Pit Mine (CM), located within the UTM facilities and from site 41 (Antas dam) in July and October 2008. We verified low pH values in water samples from CM (3.7) in comparison to the ones found at site 41 (6.65). There was a higher medium density value of Acidithiobacillus ferrooxidans, Acidithiobacillus thiooxidans and heterotrophic acidophilic bacteria in water samples at site CM compared to the values recorded from samples at site 41. Medium values of Fe{sup 2+}, uranium and zinc in samples from the site CM were higher than at site 41. The concentration of fluoride (68.5 mL{sup -l}) and manganese (2.34 mL{sup -1}) in water samples from site 41 were above the limits fixed for water bodies in Resolution CONAMA 357. The relative seasonal variation of some variables observed at site CM (low pH values, high densities of Acidithiobacillus sp. and heterotrophic acidophilic bacteria) shows that this site is one of the main sites of occurrence of acid mine drainage and action of bioleaching bacteria at UTM. (author)

  3. A novel denitrifying methanotroph of the NC10 phylum and its microcolony

    Science.gov (United States)

    He, Zhanfei; Cai, Chaoyang; Wang, Jiaqi; Xu, Xinhua; Zheng, Ping; Jetten, Mike S. M.; Hu, Baolan

    2016-09-01

    The NC10 phylum is a candidate phylum of prokaryotes and is considered important in biogeochemical cycles and evolutionary history. NC10 members are as-yet-uncultured and are difficult to enrich, and our knowledge regarding this phylum is largely limited to the first species ‘Candidatus Methylomirabilis oxyfera’ (M. oxyfera). Here, we enriched NC10 members from paddy soil and obtained a novel species of the NC10 phylum that mediates the anaerobic oxidation of methane (AOM) coupled to nitrite reduction. By comparing the new 16S rRNA gene sequences with those already in the database, this new species was found to be widely distributed in various habitats in China. Therefore, we tentatively named it ‘Candidatus Methylomirabilis sinica’ (M. sinica). Cells of M. sinica are roughly coccus-shaped (0.7–1.2 μm), distinct from M. oxyfera (rod-shaped; 0.25–0.5 × 0.8–1.1 μm). Notably, microscopic inspections revealed that M. sinica grew in honeycomb-shaped microcolonies, which was the first discovery of microcolony of the NC10 phylum. This finding opens the possibility to isolate NC10 members using microcolony-dependent isolation strategies.

  4. Isolation and characterisation of mineral-oxidising "Acidibacillus" spp. from mine sites and geothermal environments in different global locations.

    Science.gov (United States)

    Holanda, Roseanne; Hedrich, Sabrina; Ňancucheo, Ivan; Oliveira, Guilherme; Grail, Barry M; Johnson, D Barrie

    2016-09-01

    Eight strains of acidophilic bacteria, isolated from mine-impacted and geothermal sites from different parts of the world, were shown to form a distinct clade (proposed genus "Acidibacillus") within the phylum Firmicutes, well separated from the acidophilic genera Sulfobacillus and Alicyclobacillus. Two of the strains (both isolated from sites in Yellowstone National Park, USA) were moderate thermophiles that oxidised both ferrous iron and elemental sulphur, while the other six were mesophiles that also oxidised ferrous iron, but not sulphur. All eight isolates reduced ferric iron to varying degrees. The two groups shared <95% similarity of their 16S rRNA genes and were therefore considered to be distinct species: "Acidibacillus sulfuroxidans" (moderately thermophilic isolates) and "Acidibacillus ferrooxidans" (mesophilic isolates). Both species were obligate heterotrophs; none of the eight strains grew in the absence of organic carbon. "Acidibacillus" spp. were generally highly tolerant of elevated concentrations of cationic transition metals, though "A. sulfuroxidans" strains were more sensitive to some (e.g. nickel and zinc) than those of "A. ferrooxidans". Initial annotation of the genomes of two strains of "A. ferrooxidans" revealed the presence of genes (cbbL) involved in the RuBisCO pathway for CO2 assimilation and iron oxidation (rus), though with relatively low sequence identities.

  5. The δ15N and δ18O values of N2O produced during the co-oxidation of ammonia by methanotrophic bacteria

    Science.gov (United States)

    Mandernack, Kevin W.; Mills, Christopher T.; Johnson, Craig A.; Rahn, Thomas; Kinney, Chad

    2009-01-01

    In order to determine if the δ15N and δ18O values of N2O produced during co-oxidation of NH4+ by methanotrophic (methane oxidizing) bacteria can be isotopically distinguished from N2O produced either by autotrophic nitrifying or denitrifying bacteria, we conducted laboratory incubation experiments with pure cultures of methanotrophic bacteria that were provided NH4Cl as an oxidation substrate. The N2O produced during NH4+ oxidation by methanotrophic bacteria showed nitrogen isotope fractionation between NH4+ and N2O (εN2O–NH4+) of − 48 and − 55‰ for Methylomonas methanica and Methylosinus trichosporium, OB3b respectively. These large fractionations are similar to those previously measured for autotrophic nitrifying bacteria and consistent with N2O formation by multiple rate limiting steps that include NH4+oxidation by the methane monooxygenase enzyme and reduction of NO2− to N2O. Consequently, N2O formed by NH4+ oxidation via methanotrophic or autotrophic nitrifying bacteria might generally be characterized by lower δ15NN2O values than that formed by denitrificaiton, although this also depends on the variability of δ15N of available nitrogen sources (e.g., NH4+, NO3−, NO2−). Additional incubations with M. trichosporium OB3b at high and low CH4 conditions in waters of different δ18O values revealed that 19–27% of the oxygen in N2O was derived from O2 with the remainder from water. The biochemical mechanisms that could explain this amount of O2 incorporation are discussed. The δ18O of N2O formed under high CH4 conditions was ~ + 15‰ more positive than that formed under lower CH4 conditions. This enrichment resulted in part from the incorporation of O2 into N2O that was enriched in 18O due to an isotope fractionation effect of − 16.1 ± 2.0‰ and − 17.5 ± 5.4‰ associated with O2 consumption during the high and low methane concentration incubations, respectively. Therefore, N2O formed by NH4+

  6. A novel integrated biorefinery process for diesel fuel blendstock production using lipids from the methanotroph, Methylomicrobium buryatense

    Energy Technology Data Exchange (ETDEWEB)

    Dong, Tao; Fei, Qiang; Genelot, Marie; Smith, Holly; Laurens, Lieve M. L.; Watson, Michael J.; Pienkos, Philip T.

    2017-03-08

    In light of the availability of low-cost methane (CH4) derived from natural gas and biogas along with increasing concerns of the greenhouse gas emissions, the production of alternative liquid biofuels directly from CH4 is a promising approach to capturing wasted energy. A novel biorefinery concept integrating biological conversion of CH4 to microbial lipids together with lipid extraction and generation of hydrocarbon fuels is demonstrated in this study for the first time. An aerobic methanotrophic bacterium, Methylomicrobium buryatense capable of using CH4 as the sole carbon source was selected on the basis of genetic tractability, cultivation robustness, and ability to accumulate phospholipids in membranes. A maximum fatty acid content of 10% of dry cell weight was obtained in batch cultures grown in a continuous gas sparging fermentation system. Although phospholipids are not typically considered as a good feedstock for upgrading to hydrocarbon fuels, we set out to demonstrate that using a combination of novel lipid extraction methodology with advanced catalyst design, we could prove the feasibility of this approach. Up to 95% of the total fatty acids from membrane-bound phospholipids were recovered by a two-stage pretreatment method followed by hexane extraction of the aqueous hydrolysate. The upgrading of extracted lipids was then demonstrated in a hydrodeoxygeation process using palladium on silica as a catalyst. Lipid conversion in excess of 99% was achieved, with a full selectivity to hydrocarbons. The final hydrocarbon mixture is dominated by 88% pentadecane (C15H32) based on decarbonylation/decarboxylation and hydrogenation of C16 fatty acids, indicating that a biological gas-to-liquid fuel (Bio-GTL) process is technically feasible.

  7. On the relationship between methane production and oxidation by anaerobic methanotrophic communities from cold seeps of the Gulf of Mexico.

    Science.gov (United States)

    Orcutt, Beth; Samarkin, Vladimir; Boetius, Antje; Joye, Samantha

    2008-05-01

    The anaerobic oxidation of methane (AOM) in the marine subsurface is a significant sink for methane in the environment, yet our understanding of its regulation and dynamics is still incomplete. Relatively few groups of microorganisms consume methane in subsurface environments--namely the anaerobic methanotrophic archaea (ANME clades 1, 2 and 3), which are phylogenetically related to methanogenic archaea. Anaerobic oxidation of methane presumably proceeds via a 'reversed' methanogenic pathway. The ANME are generally associated with sulfate-reducing bacteria (SRB) and sulfate is the only documented final electron acceptor for AOM in marine sediments. Our comparative study explored the coupling of AOM with sulfate reduction (SR) and methane generation (MOG) in microbial communities from Gulf of Mexico cold seep sediments that were naturally enriched with methane and other hydrocarbons. These sediments harbour a variety of ANME clades and SRB. Following enrichment under an atmosphere of methane, AOM fuelled 50-100% of SR, even in sediment slurries containing petroleum-associated hydrocarbons and organic matter. In the presence of methane and sulfate, the investigated microbial communities produce methane at a small fraction ( approximately 10%) of the AOM rate. Anaerobic oxidation of methane, MOG and SR rates decreased significantly with decreasing concentration of methane, and in the presence of the SR inhibitor molybdate, but reacted differently to the MOG inhibitor 2-bromoethanesulfonate (BES). The addition of acetate, a possible breakdown product of petroleum in situ and a potential intermediate in AOM/SR syntrophy, did not suppress AOM activity; rather acetate stimulated microbial activity in oily sediment slurries.

  8. Culture scale-up and immobilisation of a mixed methanotrophic consortium for methane remediation in pilot-scale bio-filters.

    Science.gov (United States)

    Karthikeyan, Obulisamy Parthiba; Saravanan, Nadarajan; Cirés, Samuel; Alvarez-Roa, Carlos; Razaghi, Ali; Chidambarampadmavathy, Karthigeyan; Velu, Chinnathambi; Subashchandrabose, Gobalakrishnan; Heimann, Kirsten

    2017-02-01

    Robust methanotrophic consortia for methane (CH4) remediation and by-product development are presently not readily available for industrial use. In this study, a mixed methanotrophic consortium (MMC), sequentially enriched from a marine sediment, was assessed for CH4 removal efficiency and potential biomass-generated by-product development. Suitable packing material for bio-filters to support MMC biofilm establishment and growth was also evaluated. The enriched MMC removed ∼7-13% CH4 under a very high gas flow rate (2.5 L min(-1); 20-25% CH4) in continuous-stirred tank reactors (∼10 L working volume) and the biomass contained long-chain fatty acids (i.e. C16 and C18). Cultivation of the MMC on plastic bio-balls abated ∼95-97% CH4 in pilot-scale non-sterile outdoor-operated bio-filters (0.1 L min(-1); 1% CH4). Contamination by cyanobacteria had beneficial effects on treating low-level CH4, by providing additional oxygen for methane oxidation by MMC, suggesting that the co-cultivation of MMC with cyanobacterial mats does not interfere with and may actually be beneficial for remediation of CH4 and CO2 at industrial scale.

  9. Short-term variations of methane concentrations and methanotrophic activity in a coastal inlet (Eckernförde Bay, Germany)

    Science.gov (United States)

    Richner, Dominik; Niemann, Helge; Steinle, Lea; Schneider von Deimling, Jens; Urban, Peter; Hoffmann, Jasper; Schmidt, Mark; Treude, Tina; Lehmann, Moritz

    2016-04-01

    Large quantities of methane are produced in anoxic sediments of continental margins and may be liberated into the overlying water column and, potentially, into the atmosphere. However, a sequence of microbially mediated methane oxidation pathways in sediments and the water column mitigate the contribution of oceans to the atmospheric methane budget. Of particular importance are methanotrophic bacteria in the water column that mediate the aerobic oxidation of methane (MOx), and represent the final sink for methane before its release to the atmosphere where it acts as a potent greenhouse gas. However methane cycling in (aerobic) marine waters is not well constrained. Particularly little is known about spatiotemporal aspects of MOx activity and the underlying key physical, chemical and biological factors. Here we show results from our investigations on methane dynamics on very short time scales of hours to days in the Eckernförde Bay (E-Bay), a costal inlet of the Baltic Sea in northern Germany featuring seasonal bottom water hypoxia/anoxia. In autumn 2014, we observed highly spatiotemporal variations in water column methane contents and MOx activity: Anoxic bottom waters in a trough in the northern part of the bay contained extremely high methane concentrations of up to 800 nM, which sharply declined at the midwater redox interface (methane remained supersaturated with respect to the atmospheric equilibrium throughout the water column at all times). The methane decrease at the redox interface was related to highly active MOx communities consuming methane under microoxic conditions at rates of up 40 nM/d. About 12 hours later, the methane content and the extend of bottom water anoxia was much lower and MOx activity was highly reduced in the northern part but strongly elevated in the southern part of the bay. A few days later, bottom water anoxia, methane loading and MOx activity was partially re-established. In this contribution, we will discuss potential forcing

  10. Methane seep in shallow-water permeable sediment harbors high diversity of anaerobic methanotrophic communities, Elba, Italy

    Directory of Open Access Journals (Sweden)

    S Emil Ruff

    2016-03-01

    Full Text Available The anaerobic oxidation of methane (AOM is a key biogeochemical process regulating methane emission from marine sediments into the hydrosphere. AOM is largely mediated by consortia of anaerobic methanotrophic archaea (ANME and sulfate-reducing bacteria (SRB, and has mainly been investigated in deep-sea sediments. Here we studied methane seepage at four spots located at 12 m water depth in coastal, organic-carbon depleted permeable sands off the Island of Elba (Italy. We combined biogeochemical measurements, sequencing-based community analyses and in situ hybridization to investigate the microbial communities of this environment. Increased alkalinity, formation of free sulfide and nearly stoichiometric methane oxidation and sulfate reduction rates up to 200 nmol g-1 day-1 indicated the predominance of sulfate-coupled AOM. With up to 40 cm thickness the zones of AOM activity were unusually large and occurred in deeper sediment horizons (20–50 cm below seafloor as compared to diffusion-dominated deep-sea seeps, which is likely caused by advective flow of pore water due to the shallow water depth and permeability of the sands. Hydrodynamic forces also may be responsible for the substantial phylogenetic and unprecedented morphological diversity of AOM consortia inhabiting these sands, including the clades ANME-1a/b, ANME-2a/b/c, ANME-3 and their partner bacteria SEEP-SRB1a and SEEP-SRB2. High microbial dispersal, the availability of diverse energy sources and high habitat heterogeneity might explain that the emission spots shared few microbial taxa, despite their physical proximity. Although the biogeochemistry of this shallow methane seep was very different to that of deep-sea seeps, their key functional taxa were very closely related, which supports the global dispersal of key taxa and underlines strong selection by methane as the predominant energy source. Mesophilic, methane-fueled ecosystems in shallow-water permeable sediments may comprise

  11. Influence of water chemistry on the distribution of an acidophilic protozoan in an acid mine drainage system at the abandoned Green Valley coal mine, Indiana, USA

    Energy Technology Data Exchange (ETDEWEB)

    Brake, S.S.; Dannelly, H.K.; Connors, K.A.; Hasiotis, S.T. [Indiana State University, Terre Haute, IN (United States). Dept. of Geography Geology & Anthropology

    2001-07-01

    Euglena mutabilis, a benthic photosynthetic protozoan that intracellularly sequesters Fe, is variably abundant in the main effluent channel that contains acid mine drainage (AMD) discharging from the Green Valley coal mine site in western Indiana. Samples of effluent (pH 3.0-4.6) taken from the main channel and samples of contaminated stream water (pH 3.3 to 8.0) collected from an adjacent stream were analyzed to evaluate the influence of water chemistry on E. mutabilis distribution. E. mutabilis communities were restricted to areas containing unmixed effluent with the thickest (up to 3 mm) benthic communities residing in effluent containing high concentrations of total Fe (up to 12110 mg/l), SO{sub 4}(up to 2940 mg/l), Al (up to 1846 mg/l), and Cl (up to 629 mg/l). Communities were also present, but much less abundant, in areas with effluent containing lower concentrations of these same constituents. In effluent where SO{sub 4} was most highly concentrated, E. mutabilis was largely absent, suggesting that extremely high concentrations of SO{sub 4} may have an adverse effect on this potentially beneficial Fe-mediating, acidophilic protozoan.

  12. Adaptation of a mixed culture of acidophiles for a tank biooxidation of refractory gold concentrates containing a high concentration of arsenic.

    Science.gov (United States)

    Hong, Jeongsik; Silva, Rene A; Park, Jeonghyun; Lee, Eunseong; Park, Jayhyun; Kim, Hyunjung

    2016-05-01

    We adapted a mixed culture of acidophiles to high arsenic concentrations to confirm the possibility of achieving more than 70% biooxidation of refractory gold concentrates containing high arsenic (As) concentration. The biooxidation process was applied to refractory gold concentrates containing approximately 139.67 g/kg of total As in a stirred tank reactor using an adapted mixed culture of Acidithiobacillus ferrooxidans and Acidithiobacillus thiooxidans. The percentage of the biooxidation process was analyzed based on the total As removal efficiency. The As removal was monitored by inductively coupled plasma (ICP) analysis, conducted every 24 h. The results obtained with the adapted culture were compared with the percentage of biooxidation obtained with a non-adapted mixed culture of A. ferrooxidans and A. thiooxidans, and with their respective pure cultures. The percentages of biooxidation obtained during 358 h of reaction were 72.20%, 38.20%, 27.70%, and 11.45% for adapted culture, non-adapted culture, and pure cultures of A. thiooxidans and A. ferrooxidans, respectively. The adapted culture showed a peak maximum percentage of biooxidation of 77% at 120 h of reaction, confirming that it is possible to obtain biooxidation percentages over 70% in gold concentrates containing high As concentrations.

  13. Cloning and expression of acidstable, high maltose-forming, Ca2+-independent α-amylase from an acidophile Bacillus acidicola and its applicability in starch hydrolysis.

    Science.gov (United States)

    Sharma, Archana; Satyanarayana, T

    2012-05-01

    The α-amylase encoding gene from acidophilic bacterium Bacillus acidicola was cloned into pET28a(+) vector and expressed in Escherichia coli BL21 (DE3). The recombinant E. coli produced a 15-fold higher α-amylase than B. acidicola strain. The recombinant α-amylase was purified to homogeneity by one-step nickel affinity chromatography using Ni(2+)-NTA resin with molecular mass of 62 KDa. It is active in the pH range between 3.0 and 7.0 and 30 and 100 °C with optimum at pH 4.0 and 60 °C. The enzyme is Ca(2+)-independent with K (m) and k (cat) values (on soluble starch) of 1.6 mg ml(-1) and 108.7 s(-1), respectively. The α-amylase of B. acidicola is acidstable, high maltose forming and Ca(2+)-independent, and therefore, is a suitable candidate for starch hydrolysis and baking.

  14. Methods of combined bioprocessing and related microorganisms, thermophilic and/or acidophilic enzymes, and nucleic acids encoding said enzymes

    Science.gov (United States)

    Thompson, David N; Apel, William A; Thompson, Vicki S; Ward, Thomas E

    2013-07-23

    A genetically modified organism comprising: at least one nucleic acid sequence and/or at least one recombinant nucleic acid isolated from Alicyclobacillus acidocaldarius and encoding a polypeptide involved in at least partially degrading, cleaving, transporting, metabolizing, or removing polysaccharides, cellulose, lignocellulose, hemicellulose, lignin, starch, sugars, sugar oligomers, carbohydrates, complex carbohydrates, chitin, heteroxylans, glycosides, xylan-, glucan-, galactan-, or mannan-decorating groups; and at least one nucleic acid sequence and/or at least one recombinant nucleic acid encoding a polypeptide involved in fermenting sugar molecules to a product. Additionally, enzymatic and/or proteinaceous extracts may be isolated from one or more genetically modified organisms. The extracts are utilized to convert biomass into a product. Further provided are methods of converting biomass into products comprising: placing the genetically modified organism and/or enzymatic extracts thereof in fluid contact with polysaccharides, cellulose, lignocellulose, hemicellulose, lignin, starch, sugars, sugar oligomers, carbohydrates, complex carbohydrates, chitin, heteroxylans, glycosides, and/or xylan-, glucan-, galactan-, or mannan-decorating groups.

  15. Methods of combined bioprocessing and related microorganisms, thermophilic and/or acidophilic enzymes, and nucleic acids encoding said enzymes

    Energy Technology Data Exchange (ETDEWEB)

    Thompson, David N.; Apel, William A.; Thompson, Vicki S.; Ward, Thomas E.

    2016-03-22

    A genetically modified organism comprising: at least one nucleic acid sequence and/or at least one recombinant nucleic acid isolated from Alicyclobacillus acidocaldarius and encoding a polypeptide involved in at least partially degrading, cleaving, transporting, metabolizing, or removing polysaccharides, cellulose, lignocellulose, hemicellulose, lignin, starch, sugars, sugar oligomers, carbohydrates, complex carbohydrates, chitin, heteroxylans, glycosides, xylan-, glucan-, galactan-, or mannan-decorating groups; and at least one nucleic acid sequence and/or at least one recombinant nucleic acid encoding a polypeptide involved in fermenting sugar molecules to a product. Additionally, enzymatic and/or proteinaceous extracts may be isolated from one or more genetically modified organisms. The extracts are utilized to convert biomass into a product. Further provided are methods of converting biomass into products comprising: placing the genetically modified organism and/or enzymatic extracts thereof in fluid contact with polysaccharides, cellulose, lignocellulose, hemicellulose, lignin, starch, sugars, sugar oligomers, carbohydrates, complex carbohydrates, chitin, heteroxylans, glycosides, and/or xylan-, glucan-, galactan-, or mannan-decorating groups.

  16. Methods of combined bioprocessing and related microorganisms, thermophilic and/or acidophilic enzymes, and nucleic acids encoding said enzymes

    Energy Technology Data Exchange (ETDEWEB)

    Thompson, David N; Apel, William A; Thompson, Vicki S; Ward, Thomas E

    2014-04-08

    A genetically modified organism comprising: at least one nucleic acid sequence and/or at least one recombinant nucleic acid isolated from Alicyclobacillus acidocaldarius and encoding a polypeptide involved in at least partially degrading, cleaving, transporting, metabolizing, or removing polysaccharides, cellulose, lignocellulose, hemicellulose, lignin, starch, sugars, sugar oligomers, carbohydrates, complex carbohydrates, chitin, heteroxylans, glycosides, xylan-, glucan-, galactan-, or mannan-decorating groups; and at least one nucleic acid sequence and/or at least one recombinant nucleic acid encoding a polypeptide involved in fermenting sugar molecules to a product. Additionally, enzymatic and/or proteinaceous extracts may be isolated from one or more genetically modified organisms. The extracts are utilized to convert biomass into a product. Further provided are methods of converting biomass into products comprising: placing the genetically modified organism and/or enzymatic extracts thereof in fluid contact with polysaccharides, cellulose, lignocellulose, hemicellulose, lignin, starch, sugars, sugar oligomers, carbohydrates, complex carbohydrates, chitin, heteroxylans, glycosides, and/or xylan-, glucan-, galactan-, or mannan-decorating groups.

  17. Methods of combined bioprocessing and related microorganisms, thermophilic and/or acidophilic enzymes, and nucleic acids encoding said enzymes

    Energy Technology Data Exchange (ETDEWEB)

    Thompson, David N.; Apel, William A.; Thompson, Vicki S.; Ward, Thomas E.

    2017-08-15

    A genetically modified organism comprising: at least one nucleic acid sequence and/or at least one recombinant nucleic acid isolated from Alicyclobacillus acidocaldarius and encoding a polypeptide involved in at least partially degrading, cleaving, transporting, metabolizing, or removing polysaccharides, cellulose, lignocellulose, hemicellulose, lignin, starch, sugars, sugar oligomers, carbohydrates, complex carbohydrates, chitin, heteroxylans, glycosides, xylan-, glucan-, galactan-, or mannan-decorating groups; and at least one nucleic acid sequence and/or at least one recombinant nucleic acid encoding a polypeptide involved in fermenting sugar molecules to a product. Additionally, enzymatic and/or proteinaceous extracts may be isolated from one or more genetically modified organisms. The extracts are utilized to convert biomass into a product. Further provided are methods of converting biomass into products comprising: placing the genetically modified organism and/or enzymatic extracts thereof in fluid contact with polysaccharides, cellulose, lignocellulose, hemicellulose, lignin, starch, sugars, sugar oligomers, carbohydrates, complex carbohydrates, chitin, heteroxylans, glycosides, and/or xylan-, glucan-, galactan-, or mannan-decorating groups.

  18. Insights into the Quorum Sensing Regulon of the Acidophilic Acidithiobacillus ferrooxidans Revealed by Transcriptomic in the Presence of an Acyl Homoserine Lactone Superagonist Analog.

    Science.gov (United States)

    Mamani, Sigde; Moinier, Danielle; Denis, Yann; Soulère, Laurent; Queneau, Yves; Talla, Emmanuel; Bonnefoy, Violaine; Guiliani, Nicolas

    2016-01-01

    While a functional quorum sensing system has been identified in the acidophilic chemolithoautotrophic Acidithiobacillus ferrooxidans ATCC 23270(T) and shown to modulate cell adhesion to solid substrates, nothing is known about the genes it regulates. To address the question of how quorum sensing controls biofilm formation in A. ferrooxidans (T), the transcriptome of this organism in conditions in which quorum sensing response is stimulated by a synthetic superagonist AHL (N-acyl homoserine lactones) analog has been studied. First, the effect on biofilm formation of a synthetic AHL tetrazolic analog, tetrazole 9c, known for its agonistic QS activity, was assessed by fluorescence and electron microscopy. A fast adherence of A. ferrooxidans (T) cells on sulfur coupons was observed. Then, tetrazole 9c was used in DNA microarray experiments that allowed the identification of genes regulated by quorum sensing signaling, and more particularly, those involved in early biofilm formation. Interestingly, afeI gene, encoding the AHL synthase, but not the A. ferrooxidans quorum sensing transcriptional regulator AfeR encoding gene, was shown to be regulated by quorum sensing. Data indicated that quorum sensing network represents at least 4.5% (141 genes) of the ATCC 23270(T) genome of which 42.5% (60 genes) are related to biofilm formation. Finally, AfeR was shown to bind specifically to the regulatory region of the afeI gene at the level of the palindromic sequence predicted to be the AfeR binding site. Our results give new insights on the response of A. ferrooxidans to quorum sensing and on biofilm biogenesis.

  19. Differential expression of two bc1 complexes in the strict acidophilic chemolithoautotrophic bacterium Acidithiobacillus ferrooxidans suggests a model for their respective roles in iron or sulfur oxidation.

    Science.gov (United States)

    Bruscella, Patrice; Appia-Ayme, Corinne; Levicán, Gloria; Ratouchniak, Jeanine; Jedlicki, Eugenia; Holmes, David S; Bonnefoy, Violaine

    2007-01-01

    Three strains of the strict acidophilic chemolithoautotrophic Acidithiobacillus ferrooxidans, including the type strain ATCC 23270, contain a petIIABC gene cluster that encodes the three proteins, cytochrome c1, cytochrome b and a Rieske protein, that constitute a bc1 electron-transfer complex. RT-PCR and Northern blotting show that the petIIABC cluster is co-transcribed with cycA, encoding a cytochrome c belonging to the c4 family, sdrA, encoding a putative short-chain dehydrogenase, and hip, encoding a high potential iron-sulfur protein, suggesting that the six genes constitute an operon, termed the petII operon. Previous results indicated that A. ferrooxidans contains a second pet operon, termed the petI operon, which contains a gene cluster that is similarly organized except that it lacks hip. Real-time PCR and Northern blot experiments demonstrate that petI is transcribed mainly in cells grown in medium containing iron, whereas petII is transcribed in cells grown in media containing sulfur or iron. Primer extension experiments revealed possible transcription initiation sites for the petI and petII operons. A model is presented in which petI is proposed to encode the bc1 complex, functioning in the uphill flow of electrons from iron to NAD(P), whereas petII is suggested to be involved in electron transfer from sulfur (or formate) to oxygen (or ferric iron). A. ferrooxidans is the only organism, to date, to exhibit two functional bc1 complexes.

  20. Bioleaching of spent Ni-Cd batteries and phylogenetic analysis of an acidophilic strain in acidified sludge

    Institute of Scientific and Technical Information of China (English)

    ZHAO Ling; WANG Liang; YANG Dong; ZHU Nanwen

    2007-01-01

    Biohydrometallurgy is a novel method to recycle discarded batteries,in which sewage sludge is used as microorganisms and culture due to the presence of indigenous Thiobacilli.A two-step continuous flow.leaching system consisting of an acidifying reactor and a leaching reactor was introduced to achieve the bioleaching of spent nickelcadmium (Ni-Cd) batteries.The acid supematant produced in the acidifying reactor by the microorganisms with ferrous ions as the substrate was conducted into the leaching reactor to dissolve electrode materials.The efficiency of a batch treatment of batteries was examined.The results showed that the complete dissolution of two AA-sized Ni-Cd batteries with 0.6 L/d acid supernatant took about 30,20,and 35 days for Ni,Cd,and Co,respectively.But the dissolution ability of the three metals was different.Cd and Co can be leached mostly for pH below 4.0 while the complete dissolution of Ni can be achieved for pH below 2.5.Meanwhile,a strain (named Thiooxidans.WL) accounting for the reduction of pH in the acidified sludge was isolated and sequenced.It was identified to be 100% similar to Acidithiobacillus ferrooxidans strain Tf-49 based on 16S rDNA sequence analysis.The relevant phylogenetic tree constructed indicates that the strain should be classified into genus Acidithiobacillus ferrooxidans.

  1. Research Progress on Methanotrophic Bacteria in Landfills and the Reduction of Methane Emission%垃圾填埋场甲烷氧化菌及甲烷减排的研究进展

    Institute of Scientific and Technical Information of China (English)

    王晓琳; 曹爱新; 周传斌; 赵恺凝; 赵国柱

    2016-01-01

    As the main source of anthropogenic methane emission,landfills globally produce 35-69 Tg methane per year. The technology of reducing the methane emission in landfills has become a hot topic at present. Methanotrophic bacteria decomposing methane are the important biological collection to reduce atmospheric methane emissions,which is of significance in keeping the balance of the methane concentration in the atmosphere. Starting from the taxonomy and characteristics of methanotrophic bacteria,and the mechanism of its oxidizing methane, we summarized the latest research progress on the methods of studying diversity,factors affecting the activities of methanotrophic bacteria in landfills,and applications of them in the biological reduction of methane emission. Based on the prior researches,the issues in current studies of methanotrophic bacteria are also discussed. We propose comprehensive measures of utilizing the complex microbial agents of methanotrophic bacteria in landfills,providing a new thought in the research and application of reducing methane emission in landfills.%垃圾填埋场是全球最重要的人为甲烷排放源之一,其全球年甲烷释放量为35-69 Tg,垃圾填埋场甲烷减排是目前全球温室气体研究的热点。甲烷氧化菌能够氧化分解甲烷,作为减少大气甲烷排放的重要生物汇,对保持大气中甲烷浓度的平衡具有重要意义。从甲烷氧化菌的类型及其特征、甲烷氧化机理着手,介绍了多样性研究方法、填埋场中甲烷氧化菌的活性影响因素及甲烷生物减排应用等最新研究进展。在综述前人研究的基础上,探讨了目前研究的不足,提出了利用甲烷氧化菌复合微生物菌剂等综合处理措施,旨为垃圾填埋场甲烷减排的研究和应用提供新的思路。

  2. 水稻土甲烷氧化菌对镉胁迫的响应%Response of methanotrophic community structure and oxidation potential to cadmium stress in a Chinese paddy soil

    Institute of Scientific and Technical Information of China (English)

    郑勇; 郑袁明; 贺纪正

    2012-01-01

    重金属污染影响土壤微生物群落结构与活性,间接影响土壤碳(如CO2、CH4)的生物地球化学循环和全球气候变化.甲烷氧化菌氧化消耗CH4,降低大气中CH4含量,在缓解由温室气体导致的全球温暖化方面起着重要作用.本研究通过短期土壤培养实验,比较研究了不同强度重金属镉(Cd)胁迫下,水稻土中甲烷氧化菌的多度、群落组成及其氧化CH4潜势的差异.结果表明,添加Cd含量越大,水稻土氧化CH4潜势越弱,甲烷氧化菌pmoA基因拷贝数显著减少;甲烷氧化菌多度与水稻土氧化CH4潜势之间存在显著正相关关系(P< 0.001).群落组成分析发现,在相对低含量Cd(1 mg·kg-1)条件下,有新的甲烷氧化菌菌属出现,而添加较高含量Cd (10 mg·kg-1)时甲烷氧化菌种类减少.总之,Cd胁迫降低水稻土中甲烷氧化菌多样性及其氧化CH4潜势,可能导致原位CH4消耗减少,从而增加稻田CH4排放.%Heavy metal contamination has strong effect on the soil microbial community and activities, and also affects the biogeochemical cycling of soil carbon (e.g. CO2 and CH4) and global climate change. Fortunately, methane-oxidizing bacteria (methanotrophs) can consume CH4 and therefore play a critical role in the mitigation of global warming. Here, a short-term, laboratory-based incubation experiment was conducted to examine the effects of cadmium (Cd) amendments on the abundance, community composition and oxidation potential of methanotrophs in a southern China paddy soil. The results showed that there were significantly lower methanotrophic oxidation potential and abundance in the treatment of Cd amendment of 10 mg·kg-1 (high-Cd) than those in the treatment of Cd amendment of 1 mg·kg-1 (low-Cd) dry weight soil and the Control. Correlation analyses indicated that the methanotrophic oxidation potentials were significantly positively correlated with the abundance of methanotrophs (P < 0.001). For the community in rice

  3. Microbial community profiling of the Chinoike Jigoku ("Blood Pond Hell") hot spring in Beppu, Japan: isolation and characterization of Fe(III)-reducing Sulfolobus sp. strain GA1.

    Science.gov (United States)

    Masaki, Yusei; Tsutsumi, Katsutoshi; Hirano, Shin-Ichi; Okibe, Naoko

    2016-09-01

    Chinoike Jigoku ("Blood Pond Hell") is located in the hot spring town of Beppu on the southern island of Kyushu in Japan, and is the site of a red-colored acidic geothermal pond. This study aimed to investigate the microbial population composition in this extremely acidic environment and to isolate/characterize acidophilic microorganism with metal-reducing ability. Initially, PCR (using bacteria- and archaea-specific primers) of environmental DNA samples detected the presence of bacteria, but not archaea. This was followed by random sequencing analysis, confirming the presence of wide bacterial diversity at the site (123 clones derived from 18 bacterial and 1 archaeal genera), including those closely related to known autotrophic and heterotrophic acidophiles (Acidithiobacillus sp., Sulfobacillus sp., Alicyclobacillus sp.). Nevertheless, successive culture enrichment with Fe(III) under micro-aerobic conditions led to isolation of an unknown archaeal organism, Sulfolobus sp. GA1 (with 99.7% 16S rRNA gene sequence identity with Sulfolobus shibatae). Unlike many other known Sulfolobus spp., strain GA1 was shown to lack sulfur oxidation ability. Strain GA1 possessed only minor Fe(II) oxidation ability, but readily reduced Fe(III) during heterotrophic growth under micro-aerobic conditions. Strain GA1 was capable of reducing highly toxic Cr(VI) to less toxic/soluble Cr(III), demonstrating its potential utility in bioremediation of toxic metal species.

  4. Methane-Fueled Syntrophy through Extracellular Electron Transfer: Uncovering the Genomic Traits Conserved within Diverse Bacterial Partners of Anaerobic Methanotrophic Archaea

    Directory of Open Access Journals (Sweden)

    Connor T. Skennerton

    2017-08-01

    Full Text Available The anaerobic oxidation of methane by anaerobic methanotrophic (ANME archaea in syntrophic partnership with deltaproteobacterial sulfate-reducing bacteria (SRB is the primary mechanism for methane removal in ocean sediments. The mechanism of their syntrophy has been the subject of much research as traditional intermediate compounds, such as hydrogen and formate, failed to decouple the partners. Recent findings have indicated the potential for extracellular electron transfer from ANME archaea to SRB, though it is unclear how extracellular electrons are integrated into the metabolism of the SRB partner. We used metagenomics to reconstruct eight genomes from the globally distributed SEEP-SRB1 clade of ANME partner bacteria to determine what genomic features are required for syntrophy. The SEEP-SRB1 genomes contain large multiheme cytochromes that were not found in previously described free-living SRB and also lack periplasmic hydrogenases that may prevent an independent lifestyle without an extracellular source of electrons from ANME archaea. Metaproteomics revealed the expression of these cytochromes at in situ methane seep sediments from three sites along the Pacific coast of the United States. Phylogenetic analysis showed that these cytochromes appear to have been horizontally transferred from metal-respiring members of the Deltaproteobacteria such as Geobacter and may allow these syntrophic SRB to accept extracellular electrons in place of other chemical/organic electron donors.

  5. Vertical profiles of community abundance and diversity of anaerobic methanotrophic archaea (ANME) and bacteria in a simple waste landfill in north China.

    Science.gov (United States)

    Dong, Jun; Ding, Linjie; Wang, Xu; Chi, Zifang; Lei, Jiansen

    2015-03-01

    Anaerobic methane oxidation (AMO) is considered to be an important sink of CH4 in habitats as marine sediments. But, few studies focused on AMO in landfills which may be an important sink of CH4 derived from waste fermentation. To show evidence of AMO and to uncover function anaerobic methanotroph (ANME) community in landfill, different age waste samples were collected in Jinqianpu landfill located in north China. Through high-throughput sequencing, Methanomicrobiales and Methanosarcinales archaea associated with ANME and reverse methanogenic archaea of Methanosarcina and Methanobacterium were detected. Sulfate-reducing bacteria (SRB) (Desulfobulbus and Desulfococcus) which could couple with ANME-conducting AMO were also found. But, the community structure of ANME had no significant difference with depths. From the results of investigation, we can come to a conclusion that sulfate-dependent anaerobic methane oxidation (SR-DAMO) would be the dominant AMO process in the landfill, while iron-dependent anaerobic methane oxidation (M/IR-DAMO) process was weak though concentration of ferric iron was large in the landfill. Denitrification-dependent anaerobic methane oxidation (NR-DAMO) was negative because of lack of nitrate and relevant function microorganisms in the landfill. Results also indicate that CH4 mitigation would have higher potential by increasing electron acceptor contents and promoting the growth of relevant function microorganisms.

  6. The use of methanotrophic bacteria for the treatment of groundwater contaminated with trichloroethene at the US Department of Energy Kansas City Plant

    Energy Technology Data Exchange (ETDEWEB)

    Garland, S.B. II; Palumbo, A.V.; Strandberg, G.W.; Donaldson, T.L.; Farr, L.L.; Eng, W.; Little, C.D. (Oak Ridge National Lab., TN (USA); Tennessee Univ., Knoxville, TN (USA). Dept. of Microbiology; Florida State Univ., Tallahassee, FL (USA). Inst. of Molecular Biophysics)

    1989-11-01

    This study was conducted to demonstrate the technical feasibility of a trickle-filter methanotrophic bioreactor for the remediation of trichloroethene (TCE) contamination in groundwater. A bench-scale continuous-flow bioreactor was constructed and operated for several months to treat synthetic contaminated groundwater and to identify the rate of TCE degradation and the parameters that control bioreactor performance. With influent concentrations of TCE and trans-1,2-dichloroethene (DCE) of 1 mg/L each and a residence time of 50 min, approximately 50% of the TCE and 90% of the DCE were degraded in a single pass through the bioreactor. Further degradation of TCE was obtained with liquid recycle. The performance of the bench-scale bioreactor indicates that bioremediation of TCE-contaminated groundwater is technically feasible. A 3-month pilot plant project to further develop the process is estimated to cost approximately $180,000. A full-scale plant ranging in size from 50 to 700 gal/min is estimated to cost from $180,000 to $1 million to construct and from $4 to $1 per 1000 gal to operate.

  7. A Novel Framework for Quantifying past Methane Recycling by Sphagnum-Methanotroph Symbiosis Using Carbon and Hydrogen Isotope Ratios of Leaf Wax Biomarkers

    Science.gov (United States)

    Nichols, Jonathan E.; Isles, Peter D. F.; Peteet, Dorothy M.

    2014-01-01

    The concentration of atmospheric methane is strongly linked to variations in Earth's climate. Currently, we can directly reconstruct the total atmospheric concentration of methane, but not individual terms of the methane cycle. Northern wetlands, dominated by Sphagnum, are an important contributor of atmospheric methane, and we seek to understand the methane cycle in these systems. We present a novel method for quantifying the proportion of carbon Sphagnum assimilates from its methanotrophic symbionts using stable isotope ratios of leaf-wax biomarkers. Carbon isotope ratios of Sphagnum compounds are determined by two competing influences, water content and the isotope ratio of source carbon. We disentangled these effects using a combined hydrogen and carbon isotope approach. We constrained Sphagnum water content using the contrast between the hydrogen isotope ratios of Sphagnum and vascular plant biomarkers. We then used Sphagnum water content to calculate the carbon isotope ratio of Sphagnum's carbon pool. Using a mass balance equation, we calculated the proportion of recycled methane contributed to the Sphagnum carbon pool, 'PRM.' We quantified PRM in peat monoliths from three microhabitats in the Mer Bleue peatland complex. Modern studies have shown that water table depth and vegetation have strong influences on the peatland methane cycle on instrumental time scales. With this new approach, delta C-13 of Sphagnum compounds are now a useful tool for investigating the relationships among hydrology, vegetation, and methanotrophy in Sphagnum peatlands over the time scales of entire peatland sediment records, vital to our understanding of the global carbon cycle through the Late Glacial and Holocene.

  8. Identification and characterization of EctR1, a new transcriptional regulator of the ectoine biosynthesis genes in the halotolerant methanotroph Methylomicrobium alcaliphilum 20Z.

    Science.gov (United States)

    Mustakhimov, Ildar I; Reshetnikov, Alexander S; Glukhov, Anatoly S; Khmelenina, Valentina N; Kalyuzhnaya, Marina G; Trotsenko, Yuri A

    2010-01-01

    Genes encoding key enzymes of the ectoine biosynthesis pathway in the halotolerant obligate methanotroph Methylomicrobium alcaliphilum 20Z have been shown to be organized into an ectABC-ask operon. Transcription of the ect operon is initiated from two promoters, ectAp(1) and ectAp(2) (ectAp(1)p(2)), similar to the sigma(70)-dependent promoters of Escherichia coli. Upstream of the gene cluster, an open reading frame (ectR1) encoding a MarR-like transcriptional regulator was identified. Investigation of the influence of EctR1 on the activity of the ectAp(1)p(2) promoters in wild-type M. alcaliphilum 20Z and ectR1 mutant strains suggested that EctR1 is a negative regulator of the ectABC-ask operon. Purified recombinant EctR1-His(6) specifically binds as a homodimer to the putative -10 motif of the ectAp(1) promoter. The EctR1 binding site contains a pseudopalindromic sequence (TATTTAGT-GT-ACTATATA) composed of 8-bp half-sites separated by 2 bp. Transcription of the ectR1 gene is initiated from a single sigma(70)-like promoter. The location of the EctR1 binding site between the transcriptional and translational start sites of the ectR1 gene suggests that EctR1 may regulate its own expression. The data presented suggest that in Methylomicrobium alcaliphilum 20Z, EctR1-mediated control of the transcription of the ect genes is not the single mechanism for the regulation of ectoine biosynthesis.

  9. In vitro cholesterol-lowering properties of Lactobacillus plantarum AN6 isolated from aji-narezushi.

    Science.gov (United States)

    Kuda, T; Yazaki, T; Ono, M; Takahashi, H; Kimura, B

    2013-09-01

    Aji-narezushi is a traditional lactic acid-fermented fish. In this study, we screened for lactose-utilizing, acidophilic, bile-resistant and cholesterol-lowering lactic acid bacteria (LAB) from aji-narezushi for use as starter strains for fermented foods, as well as for use as probiotics. Of the 301 LAB isolates, 277 fermented lactose, and among these, 171 grew in de Man, Rogosa and Sharpe broth adjusted to pH 3·5. Thirty-four of the isolates were grown in a broth containing 3% (w/v) bile. All of the isolates were lactobacilli. Seven isolates that demonstrated cholesterol-lowering activity in ethanolic solution were selected. All of the isolates were identified as Lactobacillus plantarum. Lactobacillus plantarum AN6 showed the highest cholesterol-lowering activity. AN6 was more resistant to acid, salt and bile than the type strain NBRC15891(T). One-half of the cholesterol-lowering effect remained after boiling AN6 for 10 min. The Fourier transform infrared (FT-IR) analysis indicated that the content of cell wall polysaccharides in AN6 is higher than ones in the type strain. These results indicate that Lact. plantarum AN6 can be used as a profitable starter organism and probiotic.

  10. Effects of conservation tillage on soil photosynthetic bacteria and typeⅡ methanotrophs%保护性耕作对土壤光合细菌和Ⅱ型甲烷氧化菌的影响

    Institute of Scientific and Technical Information of China (English)

    王敬敬; 李新宇; 徐明恺; 苏振成; 李旭; 孙健; 张惠文

    2012-01-01

    保护性耕作对土壤微生物具有明显的保护效应,但是其对土壤光合细菌和甲烷氧化菌的影响却鲜有报道.本文采用土壤宏基因组16S rDNA变性梯度凝胶电泳(DGGE)和荧光定量PCR技术比较了不同耕作模式(免耕和传统翻耕)和不同秸秆覆盖量(0、50%、100%)对潮土中光合细菌和Ⅱ型甲烷氧化菌数量和群落结构的影响.结果表明:免耕土壤中光合细菌的多样性(多样性指数H=2.47)显著高于传统翻耕土壤(多样性指数H=2.35),且与土壤总氮呈显著正相关,数量略低于传统翻耕土壤;光合细菌的数量和多样性虽均随着秸秆覆盖量的增加而有所增加,但不显著;虽然免耕和秸秆覆盖对Ⅱ型甲烷氧化菌数量和多样性产生了有益的影响,但是耕作模式、秸秆覆盖及二者互作对其影响均不显著;不同处理中光合细菌和Ⅱ型甲烷氧化菌的种群结构无明显变化,光合细菌优势种群以根瘤菌目(Rhizobiales)和鞘脂单胞菌目(Sphingomonadales)为主,Ⅱ型甲烷氧化菌优势种群主要为甲基孢囊菌科(Methylocystaceae)的细菌类群.%Conservation tillage has beneficial effects on soil microbes, but the effects on soil photosynthetic,bacteria and methanotrophs are rarely reported. In this study, denaturing gel gradient electrophoresis (DGGE) and quantitative PCR technique were adopted to investigate the abundance and community structure of photosynthetic bacteria and type Ⅱ methanotrophs in a fluvo-aquic soil as affected by different tillage modes ( no-tillage and conventional tillage) and straw mulching (0, 50% , 100% ). Under no tillage, the diversity of soil photosynthetic bacteria was significantly higher but the abundance was slightly lower, as compared with those under conventional tillage, and there was a significant positive correlation between the diversity of soil photosynthetic bacteria and the soil total nitrogen. Both the abundance and the diversity of soil

  11. Unusual novel n-4 polyunsaturated fatty acids in cold-seep mussels (Bathymodiolus japonicus and Bathymodiolus platifrons), originating from symbiotic methanotrophic bacteria.

    Science.gov (United States)

    Saito, Hiroaki

    2008-07-25

    Novel fatty acids originated from the two cold-seep mussels Bathymodiolus japonicus and Bathymodiolus platifrons, which host methane-oxidizing bacteria, were determined by using gas chromatography-mass spectrometry analysis of the 4,4-dimethyloxazoline derivatives. The major polyunsaturated fatty acids (PUFAs) in the two mussels belong to unusual n-4 and n-7 methylene interrupted PUFAs, such as 18:3 n-7,10,13 (Delta5,8,11-18:3), 18:4 n-4,7,10,13 (Delta5,8,11,14-18:4), 20:3n-7,10,13 (Delta7,10,13-20:3), 20:4n-4,7,10,13 (Delta7,10,13,16-20:4), and 21:4n-7,10,13,16 (Delta5,8,11,14-20:4). The similarity of fatty acids in the two Bathymodiolus species produced by the symbiotic bacteria, indicate occurrence of highly homologous mussel symbionts. In contrast to the lipids of shallow-water mussel Mytilus galloprovincialis, which contains photosynthetic n-3 PUFAs, the two Bathymodiolus mussels were lacking in docosahexaenoic acid and icosapentaenoic acid even though they are marine animals. These findings suggest the Bathymodiolus species survive independently of photosynthetic products, similar to the Calyptogena clams, which house sulfur-oxidizing bacteria and whose lipid contains n-4 non-methylene interrupted PUFAs (20:3n-4,7,15 (Delta5,13,16-20:3), 20:4n-1,4,7,15 (Delta5,13,16,19-20:4), and 21:3n-4,7,16 (Delta5,14,17-20:3)). The similarity in n-4 fatty acids between the mussels and the clam suggests that these bivalves depend on analogous n-4 family PUFAs and that the n-4 PUFA family is a characteristic of all vent bivalves depending on geothermal energy. The differences of the n-4 PUFAs between the mussels and the clam suggest a generic specificity of symbiotic bacteria and differences in lipid physiology between thiotrophic and methanotrophic symbionts. Such a highly diversified variety of n-4 family PUFAs in the mussels and the clam under different environments presumably increase the great potential of the chemosynthetic bacteria.

  12. Miniaturized extinction culturing is the preferred strategy for rapid isolation of fast-growing methane-oxidizing bacteria.

    Science.gov (United States)

    Hoefman, Sven; van der Ha, David; De Vos, Paul; Boon, Nico; Heylen, Kim

    2012-05-01

    Methane-oxidizing bacteria (MOB) have a large potential as a microbial sink for the greenhouse gas methane as well as for biotechnological purposes. However, their application in biotechnology has so far been hampered, in part due to the relative slow growth rate of the available strains. To enable the availability of novel strains, this study compares the isolation of MOB by conventional dilution plating with miniaturized extinction culturing, both performed after an initial enrichment step. The extinction approach rendered 22 MOB isolates from four environmental samples, while no MOB could be isolated by plating. In most cases, extinction culturing immediately yielded MOB monocultures making laborious purification redundant. Both type I (Methylomonas spp.) and type II (Methylosinus sp.) MOB were isolated. The isolated methanotrophic diversity represented at least 11 different strains and several novel species based on 16S rRNA gene sequence dissimilarity. These strains possessed the particulate (100%) and soluble (64%) methane monooxygenase gene. Also, 73% of the strains could be linked to a highly active fast-growing mixed MOB community. In conclusion, miniaturized extinction culturing was more efficient in rapidly isolating numerous MOB requiring little effort and fewer materials, compared with the more widely applied plating procedure. This miniaturized approach allowed straightforward isolation and could be very useful for subsequent screening of desired characteristics, in view of their future biotechnological potential.

  13. The protein ORF80 from the acidophilic and thermophilic archaeon Sulfolobus islandicus binds highly site-specifically to double-stranded DNA and represents a novel type of basic leucine zipper protein

    Science.gov (United States)

    Lipps, Georg; Ibanez, Pablo; Stroessenreuther, Thomas; Hekimian, Katya; Krauss, Gerhard

    2001-01-01

    The cryptic high copy number plasmid pRN1 from the thermophilic and acidophilic crenarchaeote Sulfolobus islandicus shares three conserved open reading frames with other S.islandicus plasmids. One of the open reading frames, namely orf80, encodes a 9.5 kDa protein that has no homology to any characterised protein. Recombinant ORF80 purified from Escherichia coli binds to double-stranded DNA in a sequence-specific manner as suggested by EMSA experiments and DNase I footprints. Two highly symmetrical binding sites separated by ∼60 bp were found upstream of the orf80 gene. Both binding sites contain two TTAA motifs as well as other conserved bases. Fluorescence measurements show that short duplex DNAs derived from a single binding site sequence are bound with submicromolar affinity and moderate cooperativity by ORF80. On DNA fragments carrying both binding sites, a rather large protein–DNA complex is formed in a highly cooperative manner. ORF80 contains an N-terminal leucine zipper motif and a highly basic domain at its C-terminus. Compared to all known basic leucine zipper proteins the order of the domains is reversed in ORF80. ORF80 may therefore constitute a new subclass of basic leucine zipper DNA-binding proteins. PMID:11812827

  14. Rhodovastum atsumiense gen. nov., sp. nov., a phototrophic alphaproteobacterium isolated from paddy soil.

    Science.gov (United States)

    Okamura, Keiko; Hisada, Takayoshi; Kanbe, Toshio; Hiraishi, Akira

    2009-02-01

    A photoorganotrophic alphaproteobacterium designated strain G2-11(T) was isolated from submerged paddy soil. This bacterium had relatively large, oval to rod-shaped cells (2.0-3.0x3.0-10 microm). Cells were motile by means of single polar flagella. The color of phototrophically growing cultures was reddish-brown. The cell extract had absorption maxima at 375, 465, 492, 529, 592, 804, and 844 nm, indicating the presence of bacteriochlorophyll a and carotenoides of the spirilloxanthin series. Vesicular intracytoplasmic membranes were present. The main component of cellular fatty acids was C(18:1)omega7c. Ubiquinone-10 and rhodoquinone-10 were the major quinones. A 16S rRNA gene sequence analysis revealed that the isolate is closest to the acidophilic aerobic photosynthetic bacterium Acidisphaera rubrifaciens strain HS-AP3(T) (93.3% similarity). The G+C content of genomic DNA is 67.8 mol%. The name Rhodovastum atsumiense gen. nov., sp. nov. is proposed for the novel isolate. The type strain is strain G2-11(T) (=NBRC 104268(T)=KCTC 5708(T)).

  15. Metabolic engineering in methanotrophic bacteria

    Energy Technology Data Exchange (ETDEWEB)

    Kalyuzhnaya, MG; Puri, AW; Lidstrom, ME

    2015-05-01

    Methane, as natural gas or biogas, is the least expensive source of carbon for (bio)chemical synthesis. Scalable biological upgrading of this simple alkane to chemicals and fuels can bring new sustainable solutions to a number of industries with large environmental footprints, such as natural gas/petroleum production, landfills, wastewater treatment, and livestock. Microbial biocatalysis with methane as a feedstock has been pursued off and on for almost a half century, with little enduring success. Today, biological engineering and systems biology provide new opportunities for metabolic system modulation and give new optimism to the concept of a methane-based bio-industry. Here we present an overview of the most recent advances pertaining to metabolic engineering of microbial methane utilization. Some ideas concerning metabolic improvements for production of acetyl-CoA and pyruvate, two main precursors for bioconversion, are presented. We also discuss main gaps in the current knowledge of aerobic methane utilization, which must be solved in order to release the full potential of methane-based biosystems. (C) 2015 International Metabolic Engineering Society. Published by Elsevier Inc. All rights reserved.

  16. Caracterización por Ardrea de microorganismos acidófilos aislados en minas de oro de Marmato, Colombia Ardrea characterisation of acidophilic micro-organisms isolated from gold mines in Marmato, Colombia

    OpenAIRE

    Osorio E. Victor Manuel; Márquez F. Edna Judith; Márquez G. Marco

    2007-01-01

    La bio-oxidación de minerales mejora la recuperación de metales valiosos y disminuye el impacto negativo causado por los subproductos de las operaciones mineras, pero las interacciones de los microorganismos involucrados son poco conocidas. Con el objeto de avanzar en el estudio de interacciones microbianas de bacterias acidófilas nativas en cultivos mixtos, en este trabajo se utilizó la siembra en medio sólido doble capa y el análisis de enzimas de restricción del ADN ribosomal amplificado (...

  17. Isoler nu

    DEFF Research Database (Denmark)

    Andersen, N.E.; Ankerstjerne, P.; Jørgensen, B.

    Omfattende vejledning, der angiver, hvordan energiforbruget i eksisterende huse kan nedbringes ved isolering, tætning og forbedring af varmeanlægget, og hvor meget det koster. Anvisningen indeholder detaljerede eksempler på efterisolering af ydervægge, tage, gulve og vinduer. Henvender sig til...

  18. The Global Redox Responding RegB/RegA Signal Transduction System Regulates the Genes Involved in Ferrous Iron and Inorganic Sulfur Compound Oxidation of the Acidophilic Acidithiobacillus ferrooxidans

    Directory of Open Access Journals (Sweden)

    Danielle Moinier

    2017-07-01

    Full Text Available The chemical attack of ore by ferric iron and/or sulfuric acid releases valuable metals. The products of these reactions are recycled by iron and sulfur oxidizing microorganisms. These acidophilic chemolithotrophic prokaryotes, among which Acidithiobacillus ferrooxidans, grow at the expense of the energy released from the oxidation of ferrous iron and/or inorganic sulfur compounds (ISCs. In At. ferrooxidans, it has been shown that the expression of the genes encoding the proteins involved in these respiratory pathways is dependent on the electron donor and that the genes involved in iron oxidation are expressed before those responsible for ISCs oxidation when both iron and sulfur are present. Since the redox potential increases during iron oxidation but remains stable during sulfur oxidation, we have put forward the hypothesis that the global redox responding two components system RegB/RegA is involved in this regulation. To understand the mechanism of this system and its role in the regulation of the aerobic respiratory pathways in At. ferrooxidans, the binding of different forms of RegA (DNA binding domain, wild-type, unphosphorylated and phosphorylated-like forms of RegA on the regulatory region of different genes/operons involved in ferrous iron and ISC oxidation has been analyzed. We have shown that the four RegA forms are able to bind specifically the upstream region of these genes. Interestingly, the phosphorylation of RegA did not change its affinity for its cognate DNA. The transcriptional start site of these genes/operons has been determined. In most cases, the RegA binding site(s was (were located upstream from the −35 (or −24 box suggesting that RegA does not interfere with the RNA polymerase binding. Based on the results presented in this report, the role of the RegB/RegA system in the regulation of the ferrous iron and ISC oxidation pathways in At. ferrooxidans is discussed.

  19. The Global Redox Responding RegB/RegA Signal Transduction System Regulates the Genes Involved in Ferrous Iron and Inorganic Sulfur Compound Oxidation of the Acidophilic Acidithiobacillus ferrooxidans.

    Science.gov (United States)

    Moinier, Danielle; Byrne, Deborah; Amouric, Agnès; Bonnefoy, Violaine

    2017-01-01

    The chemical attack of ore by ferric iron and/or sulfuric acid releases valuable metals. The products of these reactions are recycled by iron and sulfur oxidizing microorganisms. These acidophilic chemolithotrophic prokaryotes, among which Acidithiobacillus ferrooxidans, grow at the expense of the energy released from the oxidation of ferrous iron and/or inorganic sulfur compounds (ISCs). In At. ferrooxidans, it has been shown that the expression of the genes encoding the proteins involved in these respiratory pathways is dependent on the electron donor and that the genes involved in iron oxidation are expressed before those responsible for ISCs oxidation when both iron and sulfur are present. Since the redox potential increases during iron oxidation but remains stable during sulfur oxidation, we have put forward the hypothesis that the global redox responding two components system RegB/RegA is involved in this regulation. To understand the mechanism of this system and its role in the regulation of the aerobic respiratory pathways in At. ferrooxidans, the binding of different forms of RegA (DNA binding domain, wild-type, unphosphorylated and phosphorylated-like forms of RegA) on the regulatory region of different genes/operons involved in ferrous iron and ISC oxidation has been analyzed. We have shown that the four RegA forms are able to bind specifically the upstream region of these genes. Interestingly, the phosphorylation of RegA did not change its affinity for its cognate DNA. The transcriptional start site of these genes/operons has been determined. In most cases, the RegA binding site(s) was (were) located upstream from the -35 (or -24) box suggesting that RegA does not interfere with the RNA polymerase binding. Based on the results presented in this report, the role of the RegB/RegA system in the regulation of the ferrous iron and ISC oxidation pathways in At. ferrooxidans is discussed.

  20. Isolation World

    OpenAIRE

    2012-01-01

    El trabajo de fin de grado tiene como nombre “Isolation World”, que en su traducción literal significa “Aislamiento del mundo”, un videojuego diseñado y creado desde cero en su totalidad, utilizando herramientas y conocimiento de lógica en programación que se han ido aprendiendo y desarrollando a lo largo de la carrera.

  1. Effects of inhibitors and NaCl on the oxidation of reduced inorganic sulfur compounds by a marine acidophilic, sulfur-oxidizing bacterium, Acidithiobacillus thiooxidans strain SH.

    Science.gov (United States)

    Kamimura, Kazuo; Higashino, Emi; Kanao, Tadayoshi; Sugio, Tsuyoshi

    2005-02-01

    The effect of NaCl and the pathways of the oxidation of reduced inorganic sulfur compounds were studied using resting cells and cell-free extracts of Acidithiobacillus thiooxidans strain SH. This isolate specifically requires NaCl for growth. The oxidation of sulfur and sulfite by resting cells was strongly inhibited by 2-heptyl-4-hydroxyquinoline-N-oxide. Carbonylcyanide m-chlorophenyl-hydrazone and monensin were also relatively strong inhibitors. Thiosulfate-oxidizing activity was not inhibited by these uncouplers. Valinomycin did not inhibit the oxidation of sulfur compounds. NaCl stimulated the sulfur- and sulfite-oxidizing activities in resting cells but not in cell-free extracts. The tetrathionate-oxidizing activity in resting cells was slightly stimulated by NaCl, whereas it did not influence the thiosulfate-oxidizing activity. Sulfide oxidation was biphasic, suggesting the formation of intermediate sulfur. The initial phase of sulfide oxidation was not affected by NaCl, whereas the subsequent oxidation of sulfur in the second phase was Na+-dependent. A model is proposed for the role of NaCl in the metabolism of reduced sulfur compounds in A. thiooxidans strain SH.

  2. Isolation and Identification of Alicyclobacillus with High Dipicolinic Acid and Heat Resistant Proteins from Mango Juice

    Directory of Open Access Journals (Sweden)

    Hamid Reza Akhbariyoon

    2016-10-01

    Full Text Available Background and Objectives: Microbial spoilage of juices and industries related with Alicyclobacillus are considerable international issues. This spore-forming bacterium causes changes in juices odor and taste. The isolation and identification of Alicyclobacillus contamination in juice producing and packaging industries has an essential role in the prevention and control of this type of spoilage bacterium in HACCP (Hazard analysis and critical control points manner.Materials and Methods: A thermo-acidophilic, non-pathogenic and sporeforming bacterium was isolated from mango juice. Preliminary identification of the isolates was based on morphological, biochemical and physiological properties. Identification at species level was made by PCR amplification. The influence of temperature in the range of 25-65°C in the growth of bacterium and in the range of 80-120°C in spore-resistant and heat resistant proteins was investigated and compared with other spore producing bacteria.Results and Conclusion: Phylogenetic analysis of the 16S rRNA gene sequencing indicated that the isolated strain constituted a distinct lineage in the Alicyclobacillus cluster and submitted to NCBI with access number Alicyclobacillus HRM-5 KM983424.1. The spores resisted 110°C for 3 h, and produced 28% dipicolinic acid more comparable to Bacillus licheniformis. Also they could produce 0.69 mg heat resistance protein after 1.5 h treatment in 100°C. The results showed that this strain could have biotechnological applications.Conflict of interests: The authors declare no conflict of interest.

  3. Complete Genome Sequence of Beijerinckia indica subsp. indica▿

    Science.gov (United States)

    Tamas, Ivica; Dedysh, Svetlana N.; Liesack, Werner; Stott, Matthew B.; Alam, Maqsudul; Murrell, J. Colin; Dunfield, Peter F.

    2010-01-01

    Beijerinckia indica subsp. indica is an aerobic, acidophilic, exopolysaccharide-producing, N2-fixing soil bacterium. It is a generalist chemoorganotroph that is phylogenetically closely related to facultative and obligate methanotrophs of the genera Methylocella and Methylocapsa. Here we report the full genome sequence of this bacterium. PMID:20601475

  4. The Application of Biomarker Genes for DNA/RNA-Stable Isotope Probing of Active Methanotrophs Responsible for Aerobic Methane Oxidation in Six Paddy Soils%基于核酸DNA/RNA同位素示踪技术的水稻土甲烷氧化微生物研究

    Institute of Scientific and Technical Information of China (English)

    郑燕; 贾仲君

    2016-01-01

    Rice fields are major source of atmospheric methane(CH4). However,30%~90% of CH4 produced in paddy soils is oxidized by methanotrophs before it escapes to the atmosphere. China holds the largest rice production in the world,but it remains largely unknown about the active methane oxidizers in paddy soils. In this study,soil microcosms of six paddy soil incubated with 13CH4 were constructed to assess active methanotrophs by tracing the isotopically labeled 13C-DNA/RNA. Six paddy soils collected from Yingtan City of Jiangxi Province(YT),Ziyang City of Sichuan Province(ZY),Jiaxing City of Zhejiang Province(JX),Changshu City of Jiangsu Province(CS),Yangzhou City of Jiangsu Province(YZ), and Wuchang City of Heilongjiang Province(WC),were incubated with 400 µmol-1 L labeled 13CH4 or unlabeled 12CH4 to determine aerobic methane oxidation kinetics. The destructive sampling was conducted when 400 µmol-1 L CH4 was consumed. 13C-DNA and 13C-RNA were obtained through ultracentrifugation of total DNA and RNA,respectively. Clone library of pmoA genes from 13C-DNA and 16S rRNA genes from 13C-RNA were constructed to analyze composition of active methanotrophic community. After ultracentrifugation of total DNA and RNA,the agarose gel electrophoresis of pmoA gene amplicons and methanotrophic 16S rRNA reverse transcription amplicons from the fractionated DNA and rRNA,respectively,were performed, indicating the incorporation of 13C-substrate into methanotrophs during the aerobic methane oxidation. DNA-SIP and rRNA-SIP each have their advantages. In contrast to DNA,the incorporation of labeled substrate into rRNA is much faster,and a greater unspecific background of ‘heavy’ nucleic acid was observed in ′heavy′fractions in rRNA-SIP than DNA-SIP,indicating the more efficient separation for DNA.. The separation of differentially labeled rRNA was effective,however,it was not as quantitative as for DNA. This resulted in a greater unspecific background of ‘heavy’ rRNA in

  5. Isolated sleep paralysis

    Science.gov (United States)

    Sleep paralysis - isolated; Parasomnia - isolated sleep paralysis ... Episodes of isolated sleep paralysis last from a few seconds to 1 or 2 minutes. During these episodes the person is unable to move ...

  6. Isolated patellofemoral osteoarthritis.

    NARCIS (Netherlands)

    Jonbergen, H.P. van; Poolman, R.W.; Kampen, A. van

    2010-01-01

    BACKGROUND AND PURPOSE: The optimal treatment for isolated patellofemoral osteoarthritis is unclear at present. We systematically reviewed the highest level of available evidence on the nonoperative and operative treatment of isolated patellofemoral osteoarthritis to develop an evidenced-based discu

  7. Isolated patellofemoral osteoarthritis.

    NARCIS (Netherlands)

    Jonbergen, H.P. van; Poolman, R.W.; Kampen, A. van

    2010-01-01

    BACKGROUND AND PURPOSE: The optimal treatment for isolated patellofemoral osteoarthritis is unclear at present. We systematically reviewed the highest level of available evidence on the nonoperative and operative treatment of isolated patellofemoral osteoarthritis to develop an evidenced-based

  8. IDENTIFICATION AND PHYSIOLOGICAL ASPECTS OF A NOVEL CAROTENOID-ENRICHED, METAL-RESISTANT MICROALGA ISOLATED FROM AN ACIDIC RIVER IN HUELVA (SPAIN)(1).

    Science.gov (United States)

    Garbayo, Inés; Torronteras, Rafael; Forján, Eduardo; Cuaresma, María; Casal, Carlos; Mogedas, Benito; Ruiz-Domínguez, María C; Márquez, Carmen; Vaquero, Isabel; Fuentes-Cordero, Juan L; Fuentes, Rocío; González-Del-Valle, Manuel; Vílchez, Carlos

    2012-06-01

    A heavy-metal-resistant, carotenoid-enriched novel unicellular microalga was isolated from an acidic river in Huelva, Spain. The isolated ribosomal 18S subunit rDNA sequence showed homology with known sequences from green microalgae, the closest sequence (98% homology) belonging to the genus Coccomyxa. The isolated microalga therefore was an up to now uncultured microalga. The microalga was isolated from Tinto River area (Huelva, Spain), an acidic river that exhibits very low pH (1.7-3.1) with high concentrations of sulfuric acid and heavy metals, including Fe, Cu, Mn, Ni, and Al. Electron micrographs show that the microalga contains a large chloroplast with a presence of lipid droplets, an increased number of starch bodies as well as electron-dense deposits and plastoglobules, the last observed only in iron-exposed cells. Unlike other acidophile microalgae, the isolated microalga showed high growth rates when cultivated photoautotrophycally (up to 0.6 d(-1) ) in a suitable culture medium prepared at our laboratory. The growth was shown to be iron dependent. When the microalga is grown in fluidized bed reactors, the high growth rates resulted in unexpectedly high productivities for being a microalga that naturally grows in acidic environments (0.32 g·L(-1) ·d(-1) ). The microalga also grows optimally on reduced carbon sources, including glucose and urea, and at an optimal temperature of 35°C. The alga pigment profile is particularly rich in carotenoids, especially lutein, suggesting that the microalga might have potential for antioxidant production, namely, xanthophylls.

  9. Characterization of Cytochrome 579, an Unusual Cytochrome Isolated from an Iron-Oxidizing Microbial Community▿

    Science.gov (United States)

    Singer, Steven W.; Chan, Clara S.; Zemla, Adam; VerBerkmoes, Nathan C.; Hwang, Mona; Hettich, Robert L.; Banfield, Jillian F.; Thelen, Michael P.

    2008-01-01

    A novel, soluble cytochrome with an unusual visible spectral signature at 579 nm (Cyt579) has been characterized after isolation from several different microbial biofilms collected in an extremely acidic ecosystem. Previous proteogenomic studies of an Fe(II)-oxidizing community indicated that this abundant red cytochrome could be extracted from the biofilms with dilute sulfuric acid. Here, we found that the Fe(II)-dependent reduction of Cyt579 was thermodynamically favorable at a pH of >3, raising the possibility that Cyt579 acts as an accessory protein for electron transfer. The results of transmission electron microscopy of immunogold-labeled biofilm indicated that Cyt579 is localized near the bacterial cell surface, consistent with periplasmic localization. The results of further protein analysis of Cyt579, using preparative chromatofocusing and sodium dodecyl sulfate-polyacrylamide gel electrophoresis, revealed three forms of the protein that correspond to different N-terminal truncations of the amino acid sequence. The results of intact-protein analysis corroborated the posttranslational modifications of these forms and identified a genomically uncharacterized Cyt579 variant. Homology modeling was used to predict the overall cytochrome structure and heme binding site; the positions of nine amino acid substitutions found in three Cyt579 variants all map to the surface of the protein and away from the heme group. Based on this detailed characterization of Cyt579, we propose that Cyt579 acts as an electron transfer protein, shuttling electrons derived from Fe(II) oxidation to support critical metabolic functions in the acidophilic microbial community. PMID:18469132

  10. Mutation and premating isolation.

    Science.gov (United States)

    Woodruff, R C; Thompson, J N

    2002-11-01

    While premating isolation might be traceable to different genetic mechanisms in different species, evidence supports the idea that as few as one or two genes may often be sufficient to initiate isolation. Thus, new mutation can theoretically play a key role in the process. But it has long been thought that a new isolation mutation would fail, because there would be no other individuals for the isolation-mutation-carrier to mate with. We now realize that premeiotic mutations are very common and will yield a cluster of progeny carrying the same new mutant allele. In this paper, we discuss the evidence for genetically simple premating isolation barriers and the role that clusters of an isolation mutation may play in initiating allopatric, and even sympatric, species divisions.

  11. Isolated Galaxies and Isolated Satellite Systems

    CERN Document Server

    Ann, H B; Choi, Yun-Young

    2009-01-01

    We search for isolated galaxies using a volume-limited sample of galaxies with 0.02r_{vir,nei} and \\rho <\\bar{\\rho} well segregates the CIG galaxies. We confirm the morphology conformity between the host and their satellites, which suggests importance of hydrodynamic interaction among galaxies within their virial radii in galaxy evolution.

  12. Alternativ isolering i bygninger

    DEFF Research Database (Denmark)

    Andersen, T.; Skibstrup Eriksen, S.

    Denne SBI-meddelelse beskriver resultaterne af en undersøgelse af bygninger, hvor der er anvendt alternativ isolering. Undersøgelsen omfatter termografering og fugtmåling af udvendige bygningsdele isoleret med alternativ isolering. Endvidere rapporteres resultaterne af målinger af indholdet af tu...

  13. Isolated Vascular Vertigo

    Science.gov (United States)

    2014-01-01

    Strokes in the distribution of the posterior circulation may present with vertigo, imbalance, and nystagmus. Although the vertigo due to a posterior circulation stroke is usually associated with other neurologic symptoms or signs, small infarcts involving the cerebellum or brainstem can develop vertigo without other localizing symptoms. Approximately 11% of the patients with an isolated cerebellar infarction present with isolated vertigo, nystagmus, and postural unsteadiness mimicking acute peripheral vestibular disorders. The head impulse test can differentiate acute isolated vertigo associated with cerebellar strokes (particularly within the territory of the posterior inferior cerebellar artery) from more benign disorders involving the inner ear. Acute audiovestibular loss may herald impending infarction in the territory of anterior inferior cerebellar artery. Appropriate bedside evaluation is superior to MRIs for detecting central vascular vertigo syndromes. This article reviews the keys to diagnosis of acute isolated vertigo syndrome due to posterior circulation strokes involving the brainstem and cerebellum. PMID:25328871

  14. Base isolation: Fresh insight

    Energy Technology Data Exchange (ETDEWEB)

    Shustov, V.

    1993-07-15

    The objective of the research is a further development of the engineering concept of seismic isolation. Neglecting the transient stage of seismic loading results in a widespread misjudgement: The force of resistance associated with velocity is mostly conceived as a source of damping vibrations, though it is an active force at the same time, during an earthquake type excitation. For very pliant systems such as base isolated structures with relatively low bearing stiffness and with artificially added heavy damping mechanism, the so called `damping`` force may occur even the main pushing force at an earthquake. Thus, one of the two basic pillars of the common seismic isolation philosophy, namely, the doctrine of usefulness and necessity of a strong damping mechanism, is turning out to be a self-deception, sometimes even jeopardizing the safety of structures and discrediting the very idea of seismic isolation. There is a way out: breaking with damping dependancy.

  15. Alternativ isolering i bygninger

    DEFF Research Database (Denmark)

    Hansen, Ernst Jan de Place

    2001-01-01

    Resume af rapport om målinger på alternative isoleringsmaterialer i bygninger, udgivet af Statens Byggeforskningsinstitut (SBI meddelelse 128) under Energistyrelsens udviklingsprogram "Miljø- og arbejdsmiljøvenlig isolering"...

  16. Isolated sleep paralysis

    OpenAIRE

    Sawant, Neena S.; Parkar, Shubhangi R.; Tambe, Ravindra

    2005-01-01

    Sleep paralysis (SP) is a cardinal symptom of narcolepsy. However, little is available in the literature about isolated sleep paralysis. This report discusses the case of a patient with isolated sleep paralysis who progressed from mild to severe SP over 8 years. He also restarted drinking alcohol to be able to fall asleep and allay his anxiety symptoms. The patient was taught relaxation techniques and he showed complete remission of the symptoms of SP on follow up after 8 months.

  17. Isolated sleep paralysis

    OpenAIRE

    Sawant, Neena S.; Shubhangi R Parkar; Tambe, Ravindra

    2005-01-01

    Sleep paralysis (SP) is a cardinal symptom of narcolepsy. However, little is available in the literature about isolated sleep paralysis. This report discusses the case of a patient with isolated sleep paralysis who progressed from mild to severe SP over 8 years. He also restarted drinking alcohol to be able to fall asleep and allay his anxiety symptoms. The patient was taught relaxation techniques and he showed complete remission of the symptoms of SP on follow up after 8 months.

  18. Isolated patellofemoral osteoarthritis

    OpenAIRE

    Jonbergen, H.P. van; Poolman, R. W.; Kampen, A. van

    2010-01-01

    BACKGROUND AND PURPOSE: The optimal treatment for isolated patellofemoral osteoarthritis is unclear at present. We systematically reviewed the highest level of available evidence on the nonoperative and operative treatment of isolated patellofemoral osteoarthritis to develop an evidenced-based discussion of treatment options. METHODS: A systematic computerized database search (Cochrane Database of Systematic Reviews, Cochrane Central Register of Controlled Trials, MEDLINE (PubMed), and EMBASE...

  19. Isolated Splenic Hydatid Disease

    Directory of Open Access Journals (Sweden)

    Alper Dilli

    2011-01-01

    Full Text Available Hydatid disease (HD continues to be a significant health problem in areas where animal husbandry is common but no proper veterinary control exists. The involvement of the spleen in HD is rare, and isolated splenic involvement is even less common. In this case report, we present isolated splenic HD in a 26-year-old female with complaint of abdominal pain, and we discuss some of the clinical aspects of HD. Evaluation of the patient with ultrasonography, computed tomography, and magnetic resonance imaging revealed the presence of an isolated splenic HD as a multivesicular cystic mass located near splenic hilus, measuring 12×11 cm. No other organ or system involvement could be demonstrated.

  20. Isolated Fallopian Tube Torsion

    Directory of Open Access Journals (Sweden)

    S. Kardakis

    2013-01-01

    Full Text Available Isolated torsion of the Fallopian tube is a rare gynecological cause of acute lower abdominal pain, and diagnosis is difficult. There are no pathognomonic symptoms; clinical, imaging, or laboratory findings. A preoperative ultrasound showing tubular adnexal masses of heterogeneous echogenicity with cystic component is often present. Diagnosis can rarely be made before operation, and laparoscopy is necessary to establish the diagnosis. Unfortunately, surgery often is performed too late for tube conservation. Isolated Fallopian tube torsion should be suspected in case of acute pelvic pain, and prompt intervention is necessary.

  1. Isolation of Photons

    CERN Document Server

    Wielers, M

    2001-01-01

    Photon identification in ATLAS is based mainly on the shower shapes in the calorimeters, conversion reconstruction and a track veto. In this note the additional gamma/jet separation power is evaluated if isolation is required around the shower as additional criteria.

  2. Motion of isolated bodies

    Energy Technology Data Exchange (ETDEWEB)

    Choquet-Bruhat, Yvonne [Academie des Sciences, 23 Quai de Conti, 75270 Paris Cedex 06 (France); Friedrich, Helmut [Max-Planck-Institut fuer Gravitationsphysik, Am Muehlenberg 1, 14476 Golm (Germany)

    2006-10-21

    It is shown that sufficiently smooth initial data for the Einstein-dust or the Einstein-Maxwell-dust equations with non-negative density of compact support develop into solutions representing isolated bodies in the sense that the matter field has spatially compact support and is embedded in an exterior vacuum solution.

  3. Motion of Isolated bodies

    CERN Document Server

    Choquet-Bruhat, Y; Choquet-Bruhat, Yvonne; Friedrich, Helmut

    2006-01-01

    It is shown that sufficiently smooth initial data for the Einstein-dust or the Einstein-Maxwell-dust equations with non-negative density of compact support develop into solutions representing isolated bodies in the sense that the matter field has spatially compact support and is embedded in an exterior vacuum solution.

  4. Biological Isolation Garment

    Science.gov (United States)

    1976-01-01

    A spinoff of astronaut's biological garment will allow hospital patients who are highly vulnerable to infection to leave their sterile habitats for several hours, carrying their germ free environment with them. Garments can be used in any of some 200 hospitals where isolation rooms are installed to treat leukemia.

  5. ISOLATED MOLECULES IN METALS

    NARCIS (Netherlands)

    1992-01-01

    In this paper, some results obtained on the formation of isolated molecules of composition SnOx in silver and SnFx in copper-are reviewed. Hyperfine interaction and ion beam interaction techniques were used for the identification of these molecules.

  6. ISOLATED MOLECULES IN METALS

    NARCIS (Netherlands)

    1992-01-01

    In this paper, some results obtained on the formation of isolated molecules of composition SnOx in silver and SnFx in copper-are reviewed. Hyperfine interaction and ion beam interaction techniques were used for the identification of these molecules.

  7. Isolated lingual leishmaniasis

    Directory of Open Access Journals (Sweden)

    Habibzadeh F

    2005-01-01

    Full Text Available Cutaneous leishmaniasis is endemic in Fars Province, southern Iran. However, mucosal leishmaniasis is extremely uncommon. Herein, we report a patient with isolated lingual leishmaniasis in an immunocompetent 40-year-old man. The lesion was totally excised. The patient was cured completely and is doing well after four years of follow-up, with no medical treatment

  8. Isolated jejunal varices.

    Directory of Open Access Journals (Sweden)

    Bhagwat S

    1995-04-01

    Full Text Available Isolated jejunal varices are an uncommon manifestation of portal hypertension. A one and a half year old boy presented with recurrent, massive gastrointestinal bleeding from jejunal varices. The bleeding site was identified at exploratory laparotomy. Jejunal resection and anastomosis resulted in complete resolution of the bleeding and there has been no recurrent bleeding over an eight month follow-up period.

  9. Proteoglycan isolation and analysis

    DEFF Research Database (Denmark)

    Woods, A; Couchman, J R

    2001-01-01

    Proteoglycans can be difficult molecules to isolate and analyze due to large mass, charge, and tendency to aggregate or form macromolecular complexes. This unit describes detailed methods for purification of matrix, cell surface, and cytoskeleton-linked proteoglycans. Methods for analysis...

  10. Isolating Triggered Star Formation

    Energy Technology Data Exchange (ETDEWEB)

    Barton, Elizabeth J.; Arnold, Jacob A.; /UC, Irvine; Zentner, Andrew R.; /KICP, Chicago /Chicago U., EFI; Bullock, James S.; /UC, Irvine; Wechsler, Risa H.; /KIPAC, Menlo

    2007-09-12

    Galaxy pairs provide a potentially powerful means of studying triggered star formation from galaxy interactions. We use a large cosmological N-body simulation coupled with a well-tested semi-analytic substructure model to demonstrate that the majority of galaxies in close pairs reside within cluster or group-size halos and therefore represent a biased population, poorly suited for direct comparison to 'field' galaxies. Thus, the frequent observation that some types of galaxies in pairs have redder colors than 'field' galaxies is primarily a selection effect. We use our simulations to devise a means to select galaxy pairs that are isolated in their dark matter halos with respect to other massive subhalos (N= 2 halos) and to select a control sample of isolated galaxies (N= 1 halos) for comparison. We then apply these selection criteria to a volume-limited subset of the 2dF Galaxy Redshift Survey with M{sub B,j} {le} -19 and obtain the first clean measure of the typical fraction of galaxies affected by triggered star formation and the average elevation in the star formation rate. We find that 24% (30.5 %) of these L* and sub-L* galaxies in isolated 50 (30) h{sup -1} kpc pairs exhibit star formation that is boosted by a factor of {approx}> 5 above their average past value, while only 10% of isolated galaxies in the control sample show this level of enhancement. Thus, 14% (20 %) of the galaxies in these close pairs show clear triggered star formation. Our orbit models suggest that 12% (16%) of 50 (30) h{sup -1} kpc close pairs that are isolated according to our definition have had a close ({le} 30 h{sup -1} kpc) pass within the last Gyr. Thus, the data are broadly consistent with a scenario in which most or all close passes of isolated pairs result in triggered star formation. The isolation criteria we develop provide a means to constrain star formation and feedback prescriptions in hydrodynamic simulations and a very general method of understanding

  11. Offset trench isolation

    Science.gov (United States)

    Roth, S. S.; Cooper, K. J.; Ray, W.; Kirsch, H. C.; Grove, Charlotte

    1994-08-01

    Feasibility of a new, recessed isolation technique that utilizes an offset, shallow trench in combination with thermal oxidation for achieving near zero final encroachment with excellent planarity is demonstrated. Etch of the shallow trench is offset from the original hardmask by an oxide sidewall spacer. After trench etch, HF is used to remove the hardmask oxide and sidewall spacers and to form a cavity which is self-aligned to the nitride edge. Exposed silicon regions are then reoxidized and encapsulated with polysilicon. Field oxide is then grown. The final field oxide profile exhibits steep sidewall angles without inducing substrate defects as evidenced by low diode leakage. Other isolation sensitive device parametrics such as gate oxide quality and metal oxide semiconductor field effect transistor threshold voltage stability are presented and exhibit good characteristics.

  12. Idiopathic isolated orbicularis weakness

    Science.gov (United States)

    MacVie, O P; Majid, M A; Husssin, H M; Ung, T; Manners, R M; Ormerod, I; Pawade, J; Harrad, R A

    2012-01-01

    Purpose Orbicularis weakness is commonly associated with seventh nerve palsy or neuromuscular and myopathic conditions such as myotonic dystrophy and myasethenia gravis. We report four cases of idiopathic isolated orbicularis weakness. Methods All four cases were female and the presenting symptoms of ocular irritation and epiphora had been present for over 7 years in three patients. All patients had lagophthalmos and three had ectropion. Three patients underwent full investigations which excluded known causes of orbicularis weakness. Two patients underwent oribularis oculi muscle biopsy and histological confirmation of orbicularis atrophy. Results All patients underwent surgery to specifically address the orbicularis weakness with satisfactory outcomes and alleviation of symptoms in all cases. Isolated orbicularis weakness may be a relatively common entity that is frequently overlooked. Conclusion Early recognition of this condition may lead to better management and prevent patients undergoing unnecessary surgical procedures. PMID:22322997

  13. So close, so different: geothermal flux shapes divergent soil microbial communities at neighbouring sites.

    Science.gov (United States)

    Gagliano, A L; Tagliavia, M; D'Alessandro, W; Franzetti, A; Parello, F; Quatrini, P

    2016-03-01

    This study is focused on the (micro)biogeochemical features of two close geothermal sites (FAV1 and FAV2), both selected at the main exhalative area of Pantelleria Island, Italy. A previous biogeochemical survey revealed high CH4 consumption and the presence of a diverse community of methanotrophs at FAV2 site, whereas the close site FAV1 was apparently devoid of methanotrophs and recorded no CH4 consumption. Next-Generation Sequencing (NGS) techniques were applied to describe the bacterial and archaeal communities which have been linked to the physicochemical conditions and the geothermal sources of energy available at the two sites. Both sites are dominated by Bacteria and host a negligible component of ammonia-oxidizing Archaea (phylum Thaumarchaeota). The FAV2 bacterial community is characterized by an extraordinary diversity of methanotrophs, with 40% of the sequences assigned to Methylocaldum, Methylobacter (Gammaproteobacteria) and Bejerickia (Alphaproteobacteria); conversely, a community of thermo-acidophilic chemolithotrophs (Acidithiobacillus, Nitrosococcus) or putative chemolithotrophs (Ktedonobacter) dominates the FAV1 community, in the absence of methanotrophs. Since physical andchemical factors of FAV1, such as temperature and pH, cannot be considered limiting for methanotrophy, it is hypothesized that the main limiting factor for methanotrophs could be high NH4(+) concentration. At the same time, abundant availability of NH4(+) and other high energy electron donors and acceptors determined by the hydrothermal flux in this site create more energetically favourable conditions for chemolithotrophs that outcompete methanotrophs in non-nitrogen-limited soils.

  14. Isolated Pulmonary Valve Endocarditis

    Directory of Open Access Journals (Sweden)

    Mohammad Ali Hatamizadeh

    2009-06-01

    Full Text Available Infective endocarditis is one of the most severe complications of parenteral drug abuse. The outstanding clinical feature of infective endocarditis in intravenous drug abusers is the high incidence of right-sided valve infection, and the tricuspid valve is involved in 60% to 70% of the cases. We herein report a case of isolated pulmonic valve infective endocarditis with a native pulmonary valve.

  15. Isolated unilateral disk edema

    OpenAIRE

    Varner P

    2011-01-01

    Paul VarnerJohn J Pershing VAMC, Poplar Bluff, MO, USAAbstract: Isolated unilateral disk edema is a familiar clinical presentation with myriad associations. Related, non-consensus terminology is a barrier to understanding a common pathogenesis. Mechanisms for the development of disk edema are reviewed, and a new framework for clinical differentiation of medical associations is presented.Keywords: disk edema, axoplasmic flow, clinical multiplier, optic neuritis, ischemic optic neuropathy, papi...

  16. Isolated unilateral disk edema

    Directory of Open Access Journals (Sweden)

    Varner P

    2011-07-01

    Full Text Available Paul VarnerJohn J Pershing VAMC, Poplar Bluff, MO, USAAbstract: Isolated unilateral disk edema is a familiar clinical presentation with myriad associations. Related, non-consensus terminology is a barrier to understanding a common pathogenesis. Mechanisms for the development of disk edema are reviewed, and a new framework for clinical differentiation of medical associations is presented.Keywords: disk edema, axoplasmic flow, clinical multiplier, optic neuritis, ischemic optic neuropathy, papilledema

  17. Isolating Triggered Star Formation

    CERN Document Server

    Barton, Elizabeth J; Zentner, Andrew R; Bullock, James S; Wechsler, Risa H

    2007-01-01

    Galaxy pairs provide a potentially powerful means of studying triggered star formation from galaxy interactions. We use a large cosmological N-body simulation coupled with a well-tested semi-analytic substructure model to demonstrate that the majority of galaxies in close pairs reside within cluster or group-size halos and therefore represent a biased population, poorly suited for direct comparison to ``field'' galaxies. Thus, the frequent observation that some types of galaxies in pairs have redder colors than ``field'' galaxies is primarily a selection effect. We select galaxy pairs that are isolated in their dark matter halos with respect to other massive subhalos (N=2 halos) and a control sample of isolated galaxies (N=1 halos) for comparison. We then apply these selection criteria to a volume-limited subset of the 2dF Galaxy Redshift Survey with M_Bj ~ 5 above their average past value, while only 10% of isolated galaxies in the control sample show this level of enhancement. Thus, 14% (20 %) of the galaxi...

  18. Isolation by distance, web service

    OpenAIRE

    Bohonak Andrew J; Jensen Jeffrey L; Kelley Scott T

    2005-01-01

    Abstract Background The population genetic pattern known as "isolation by distance" results from spatially limited gene flow and is a commonly observed phenomenon in natural populations. However, few software programs exist for estimating the degree of isolation by distance among populations, and they tend not to be user-friendly. Results We have created Isolation by Distance Web Service (IBDWS) a user-friendly web interface for determining patterns of isolation by distance. Using this site, ...

  19. ADRENALECTOMY FOR ISOLATED METASTASES

    Directory of Open Access Journals (Sweden)

    Svetlana Shumarova

    2017-08-01

    Full Text Available Background: Isolated adrenal gland metastases are not frequently finding. The aim of the present retrospective study was to estimate clinical and pathological parameters that could be used to predict survival after adrenalectomy. Material and methods: A total of 34 patients with adrenal gland masses suspected to be metastases was included in this study. The group of patients with isolated metastases was19 (56% and a group of patients with adrenal adenomas -15 (44%. Results: The sample of patients consisted of 18 (53% men and 16 (47% women from 40 to 81 years old with a mean (±SD age of 61.6±10.3 years, presented with adrenal mass suspected to be metastases. Nineteen (56% of them had demonstrated metastases and 16 (84% metachronous with median overall survival (OS 54.6 (range 43-66 months. Median OS in the group with metastases was 22.6 months. Lung carcinoma was the most common primary tumour metastasizing in the adrenal gland – 58% of all metastases with a disease-free interval (DFI of 13 months. It was presented by shorter median survival than the rest primary tumour types (37.8 vs. 96.7months; log-rank test, p=0.028. In the multivariate Cox’s hazard analysis of the surgical technique was found to be an independent prognostic factor (p=0.047, together with lung carcinoma vs. renal cell carcinoma (p=0.045. Conclusion: Adrenalectomy due to isolated metastases in the adrenal glands showed the median overall survival of 22.6 months. Shorter survival periods were associated with lung carcinoma, DFI<12months, conversion to open surgery, synchronous metastases, but not with age, tumour size or resection status.

  20. NORMOLIPEMIC ISOLATED XANTHOMA

    Directory of Open Access Journals (Sweden)

    Gowri Sankar

    2013-10-01

    Full Text Available ABSTRACT: Xanthomas are often a manifestation of underlying lipid abnormalities.A 17 - year - old female presented to our hospital with pain and swelling over t he left knee joint.Routine investigations and lipid profile were normal. Dermatological examination and Psychiatric examination were done and found to be normal. Histopathology from the lesion revealed macrophages and Touton type of giant cells. We present a case of normolipemic isolated xanthoma, which not only is an an uncommon occurrence but also needs a regular follow up because a high association with reticuloendothelial malignancies has also been reported.

  1. Journey from isolation

    Directory of Open Access Journals (Sweden)

    Wim A. Dreyer

    2011-06-01

    Full Text Available Since the Ottawa Consultation in 1982, the relationship between the Nederduitsch Hervormde Kerk van Afrika (NHKA and the World Alliance of Reformed Churches (WARC was nonexistent. In the NHKA it became progressively clear that it would be impossible to travel the road of faith alone. This article examined the factors which contributed to the growing isolation of the NHKA, especially nationalism, a particularistic ecclesiology and the rejection of Apartheid by international ecumenical bodies. It also reflected on efforts of the NHKA to return to the international ecumenical movement.

  2. Diabetes in population isolates

    DEFF Research Database (Denmark)

    Grarup, Niels; Moltke, Ida; Albrechtsen, Anders

    2015-01-01

    Type 2 diabetes (T2D) is an increasing health problem worldwide with particularly high occurrence in specific subpopulations and ancestry groups. The high prevalence of T2D is caused both by changes in lifestyle and genetic predisposition. A large number of studies have sought to identify...... on glucose-stimulated plasma glucose, serum insulin levels, and T2D. The variant defines a specific subtype of non-autoimmune diabetes characterized by decreased post-prandial glucose uptake and muscular insulin resistance. These and other recent findings in population isolates illustrate the value...

  3. Isolation of RNP granules

    DEFF Research Database (Denmark)

    Jønson, Lars; Nielsen, Finn Cilius; Christiansen, Jan

    2011-01-01

    be regarded as a supramolecular assembly of RNA and protein, probably representing several overlapping post-transcriptional operons. The present protocol describes how RNP granules may be isolated by the transgenic expression of a 3X FLAG version of an RNA-binding protein under tetracycline control via...... the tetracycline receptor/operator complex. In this way, inclusion of an appropriate tetracycline concentration ensures expression of the tagged version at the endogenous level, and the 3X FLAG tag is a convenient "handle" for the subsequent immunoprecipitation by immobilized anti-FLAG antibody....

  4. [Isolated Ledderhose fibromatosis plantaris].

    Science.gov (United States)

    Runkel, N; Göhring, U; Friedl, W; Roeren, T

    1993-07-01

    Plantar fibromatosis or Ledderhose syndrome has rarely been discussed in the medical literature. This clinical entity includes nodular Dupuytren-like indurations of the plantar aponeurosis. We present a case of a young man with isolated disease of his right foot and describe the clinical and pathomorphological features of this disease as well as its characteristic findings at magnetic resonance tomography. The therapy of choice is a subtotal resection of the plantar aponeurosis to prevent irreversible contractions of the toes. The prognosis following fascietomy is good.

  5. Isolation of Mouse Neutrophils.

    Science.gov (United States)

    Swamydas, Muthulekha; Luo, Yi; Dorf, Martin E; Lionakis, Michail S

    2015-08-03

    Neutrophils represent the first line of defense against bacterial and fungal pathogens. Indeed, patients with inherited and acquired qualitative and quantitative neutrophil defects are at high risk for developing bacterial and fungal infections and suffering adverse outcomes from these infections. Therefore, research aiming at defining the molecular factors that modulate neutrophil effector function under homeostatic conditions and during infection is essential for devising strategies to augment neutrophil function and improve the outcome of infected individuals. This unit describes a reproducible density gradient centrifugation-based protocol that can be applied in any laboratory to harvest large numbers of highly enriched and highly viable neutrophils from the bone marrow of mice both at the steady state and following infection with Candida albicans as described in UNIT. In another protocol, we also present a method that combines gentle enzymatic tissue digestion with a positive immunomagnetic selection technique or Fluorescence-activated cell sorting (FACS) to harvest highly pure and highly viable preparations of neutrophils directly from mouse tissues such as the kidney, the liver or the spleen. Finally, methods for isolating neutrophils from mouse peritoneal fluid and peripheral blood are included. Mouse neutrophils isolated by these protocols can be used for examining several aspects of cellular function ex vivo including pathogen binding, phagocytosis and killing, neutrophil chemotaxis, oxidative burst, degranulation and cytokine production, and for performing neutrophil adoptive transfer experiments.

  6. Schizosaccharomyces isolation method

    Directory of Open Access Journals (Sweden)

    Benito Santiago

    2014-01-01

    Full Text Available This study discusses the optimization of a selective and differential medium which would facilitate the isolation of Schizosaccharomyces (a genus with a low incidence compared to other microorganisms to select individuals from this genus for industrial purposes, especially in light of the recent recommendation of the use of yeasts from this genus in the wine industry by the International Organisation of Vine and Wine, or to detect the presence of such yeasts, for those many authors who consider them food spoilers. To this end, we studied various selective differential agents based on the main physiological characteristics of these species, such as their high resistances to high concentrations of sugar, sulfur dioxide, sorbic acid, benzoic acid, acetic acid or malo ethanolic fermentation. This selective medium is based on the genus resistance to the antibiotic actidione and its high resistance to inhibitory agents such as benzoic acid. Malic acid was used as a differential factor due to the ability of this genus to metabolise it to ethanol, which allows detecting of the degradation of this compound. Lastly, the medium was successfully used to isolate strains of Schizosaccharomyces pombe from honey and honeycombs.

  7. Isolated hepatosplenic tuberculosis

    Institute of Scientific and Technical Information of China (English)

    Vinay Kumar; Durgatosh Pandey

    2008-01-01

    BACKGROUND:Tuberculosis rarely involves the liver and spleen and when it does so, it is usually associated with disseminated disease. METHOD:We report a patient with isolated tuberculosis of the liver and spleen. RESULTS:A 50-year-old man presented with weakness and loss of weight for two months. CT scan of the abdomen showed multiple small hypodense lesions in the liver and spleen. Image-guided biopsy of liver lesion was inconclusive as it showed normal hepatocytes with interspersed lymphocytes. On laparotomy, the liver and spleen were studded with multiple small nodules. A 5-cm wedge of the liver was resected. Histopathological examination conifrmed the diagnosis of tubercular hepatitis. A detailed work-up failed to identify any other focus of tuberculosis. CONCLUSION:Although rare, tuberculosis should also be considered in the differential diagnosis for multiple nodules in the liver and spleen, along with lymphoproliferative diseases and metastatic lesions.

  8. Clinically isolated laryngeal sarcoidosis

    DEFF Research Database (Denmark)

    Plaschke, Christina Caroline; Owen, Hanne Hoejris; Rasmussen, Niels

    2011-01-01

    Laryngeal sarcoidosis is rare (0.5% of patients with sarcoidosis), the pathogenesis is unknown and the optimal treatment remains a matter of debate. We undertook this study to elucidate possible pathogenic factors in clinically isolated laryngeal sarcoidosis and to describe results...... of supraglottoplastic surgery. From 1995 to 2009, we identified six patients with histologically proven sarcoidosis of the larynx treated at Rigshospitalet. All patients were subjected to a panel of blood tests and MR scan of the head and neck. All patients had dyspnoea at admission, and five were subjected...... successful to maintain normal breathing. None of the many parameters examined--some previously having been found to be abnormal in sarcoidosis--were abnormal in the present cohort. We are therefore unable to elucidate the pathogenesis. The combined surgical approach re-established normal airway function...

  9. Isolated sulfite oxidase deficiency.

    Science.gov (United States)

    Relinque, B; Bardallo, L; Granero, M; Jiménez, P J; Luna, S

    2015-03-10

    Sulfite oxidase deficiency is an uncommon metabolic disease. Only few cases of its isolated form have been reported in the literature. We report a case of severe neonatal onset. A newborn baby of 41 weeks gestational age, weighted at birth of 3240 grams and had an Apgar score of 6-10-10. Fifty-three hours after being born, the baby started with seizures that were refractory to antiepileptic treatment. Brain function was monitored using a-EEG. Laboratory and imaging tests were performed. All of them were consistent with sulfite oxidase deficiency. The diagnosis was confirmed by genetic testing. We highlight the importance of this disease as part of the differential diagnosis of seizures during the neonatal period, as well as the importance of the therapeutic support based on dietary restrictions. It's also remarkable the possibility of prenatal diagnosis by quantifying enzyme activity and it's also possible carrying out DNA mutational analysis.

  10. Patients experience of source isolation

    DEFF Research Database (Denmark)

    Johansen, Kamilla; Pedersen, Didde; Kragbak, Nina

    2013-01-01

    Patients´ experience of source isolation - a literature review Title: Patients experience of source isolation – a literature review Authors: Nina Kragbak*, Didde Pedersen*, Kamilla Johansen* and Peter E. Jensen** *students, **lecturer, bachelor in science and Ph.D., at VIA University College......, Nursing education in Århus, Hedeager 2, 8200 Aarhus N, Denmark. Background: Medical treatment and care of patients with infections may include source isolation of the patient, to avoid spreading of the infection. However, isolation is a potential physiological and psychological stress factor...... of the patients perspectives of being isolated to identify areas of potential interest for developing new caring strategies to minimize the negative side effects of isolation. Methods: Literature was systematically searched in CINAHL, Nursing Reference Center, Social Care Online, SveMed+, The Cochrane Library...

  11. Protein loss during nuclear isolation

    OpenAIRE

    1983-01-01

    Cryomicrodissection makes possible the measurement of the entire in vivo protein content of the amphibian oocyte nucleus and provides a heretofore missing baseline for estimating protein loss during nuclear isolation by other methods. When oocyte nuclei are isolated into an aqueous medium, they lose 95% of their protein with a half-time of 250 s. This result implies an even more rapid loss of protein from aqueously isolated nuclei of ordinary-size cells.

  12. A hybrid base isolation system

    Energy Technology Data Exchange (ETDEWEB)

    Hart, G.C. [Univ. of California, Los Angeles, CA (United States); Lobo, R.F.; Srinivasan, M. [Hart Consultant Group, Santa Monica, CA (United States); Asher, J.W. [kpff Engineers, Santa Monica, CA (United States)

    1995-12-01

    This paper proposes a new analysis procedure for hybrid base isolation buildings when considering the displacement response of a base isolated building to wind loads. The system is considered hybrid because of the presence of viscous dampers in the building above the isolator level. The proposed analysis approach incorporates a detailed site specific wind study combined with a dynamic nonlinear analysis of the building response.

  13. Isolation and the parish ministry

    OpenAIRE

    Irvine, Andrew R.

    1989-01-01

    The purpose of this thesis was to examine the concept of isolation as it occurs within the profession of ministry. Isolation, for the purpose of this thesis, is defined social-psychologically. Within the field research isolation is considered as evidenced professionally, socially and spiritually. This study utilized as its sample base 200 hundred Church of Scotland ministers (15% of total population) which provided 159 usable responses to an extensive mail survey. The mai...

  14. Electricity generation from tetrathionate in microbial fuel cells by acidophiles.

    Science.gov (United States)

    Sulonen, Mira L K; Kokko, Marika E; Lakaniemi, Aino-Maija; Puhakka, Jaakko A

    2015-03-02

    Inorganic sulfur compounds, such as tetrathionate, are often present in mining process and waste waters. The biodegradation of tetrathionate was studied under acidic conditions in aerobic batch cultivations and in anaerobic anodes of two-chamber flow-through microbial fuel cells (MFCs). All four cultures originating from biohydrometallurgical process waters from multimetal ore heap bioleaching oxidized tetrathionate aerobically at pH below 3 with sulfate as the main soluble metabolite. In addition, all cultures generated electricity from tetrathionate in MFCs at pH below 2.5 with ferric iron as the terminal cathodic electron acceptor. The maximum current and power densities during MFC operation and in the performance analysis were 79.6 mA m(-2) and 13.9 mW m(-2) and 433 mA m(-2) and 17.6 mW m(-2), respectively. However, the low coulombic efficiency (below 5%) indicates that most of the electrons were directed to other processes, such as aerobic oxidation of tetrathionate and unmeasured intermediates. The microbial community analysis revealed that the dominant species both in the anolyte and on the anode electrode surface of the MFCs were Acidithiobacillus spp. and Ferroplasma spp. This study provides a proof of concept that tetrathionate serves as electron donor for biological electricity production in the pH range of 1.2-2.5. Copyright © 2014 Elsevier B.V. All rights reserved.

  15. Functional genomics of the thermo-acidophilic archaeon Sulfolobus solfataricus

    NARCIS (Netherlands)

    Oost, van der J.; Walther, J.; Brouns, S.J.J.; Werken, van de H.J.G.; Snijders, A.P.L.; Wright, P.C.; Andersson, A.; Bernander, R.; Vos, de W.M.

    2006-01-01

    Archaea and bacteria that optimally grow at temperatures above 60C and 80C are referred to as thermophiles and hyperthermophiles, respectively (Stetter, 1996). Since their discovery in the late 1960s (Brock and Freeze, 1969), attempts were made to reveal the secrets of the thermal resistance of thes

  16. Electricity generation from tetrathionate in microbial fuel cells by acidophiles

    Energy Technology Data Exchange (ETDEWEB)

    Sulonen, Mira L.K., E-mail: mira.sulonen@tut.fi; Kokko, Marika E.; Lakaniemi, Aino-Maija; Puhakka, Jaakko A.

    2015-03-02

    Highlights: • Electricity can be generated from tetrathionate in MFCs at pH below 2.5. • Tetrathionate disproportionated to sulfate and elemental sulfur. • Biohydrometallurgical process waters contained electrochemically active bacteria. • Acidithiobacillus spp. and Ferroplasma spp. were identified from the MFCs. - Abstract: Inorganic sulfur compounds, such as tetrathionate, are often present in mining process and waste waters. The biodegradation of tetrathionate was studied under acidic conditions in aerobic batch cultivations and in anaerobic anodes of two-chamber flow-through microbial fuel cells (MFCs). All four cultures originating from biohydrometallurgical process waters from multimetal ore heap bioleaching oxidized tetrathionate aerobically at pH below 3 with sulfate as the main soluble metabolite. In addition, all cultures generated electricity from tetrathionate in MFCs at pH below 2.5 with ferric iron as the terminal cathodic electron acceptor. The maximum current and power densities during MFC operation and in the performance analysis were 79.6 mA m{sup −2} and 13.9 mW m{sup −2} and 433 mA m{sup −2} and 17.6 mW m{sup −2}, respectively. However, the low coulombic efficiency (below 5%) indicates that most of the electrons were directed to other processes, such as aerobic oxidation of tetrathionate and unmeasured intermediates. The microbial community analysis revealed that the dominant species both in the anolyte and on the anode electrode surface of the MFCs were Acidithiobacillus spp. and Ferroplasma spp. This study provides a proof of concept that tetrathionate serves as electron donor for biological electricity production in the pH range of 1.2–2.5.

  17. 利用嗜酸混合菌从尼日利亚闪锌矿和方铅矿中生物浸出Zn(Ⅱ)和Pb(Ⅱ)%Bioleaching of Zn(Ⅱ) and Pb(Ⅱ) from Nigerian sphalerite and galena ores by mixed culture of acidophilic bacteria

    Institute of Scientific and Technical Information of China (English)

    AlafaraA.BABA; Folahan A.ADEKOLA; Rasaq F.ATATA; Risikat N.AHMED; Sandeep PANDA4

    2011-01-01

    Zn(Ⅱ) and Pb(Ⅱ) from Nigerian sphalerite and galena ores were bioleached by a mixed culture of acidophilic bacteria.The influences ofpH and ferric ion on the bioleaching rates of sphalerite and galena were examined.The result shows that pH 2.1 and 2.7 are favourable for the leaching of Zn(Ⅱ) and Pb(Ⅱ) from sphalerite and galena,respectively.It was observed that the use of agarose-simulated media caused cells to excrete exopolymers containing ferric ions which enhanced oxidation.The oxidation equilibrium for sphalerite and galena took 3 and 4 d,respectively.About 38.3% sphalerite and 34.2% galena were leached within 1 d and approximately 92.0% Zn(Ⅱ) and 89.0% Pb(Ⅱ) were recovered in 5 d,respectively.The unleached residual products were examined by X-ray diffraction for sphalerite,revealing the presence of elemental sulphur(S),zinc sulphate (ZnSO4) and few traces of calcium aluminate (Ca3Al2O6).The XRD pattern also indicates the presence of elemental sulphur (S),lead sulphate (PbSO4) and few traces ofitoite [Pb(S,Ge)(O,OH)4] and cobalt lead silicate [Pb8C0(Si2O7)3] in the urdeached galena ore.%利用嗜酸混合菌在尼日利亚闪锌矿和方铅矿中生物浸出Zn(Ⅱ)和Pb(Ⅱ),研究pH值和铁离子对闪锌矿和方铅矿浸出率的影响.结果表明:闪锌矿和方铅矿中Zn(Ⅱ)和Pb(Ⅱ)的氧化物在pH值分别为2.1和2.7时浸出效果最好.用模拟的琼脂糖培养基使细胞分泌出含铁离子的外聚合物,以此加强氧化.闪锌矿和方铅矿达到氧化平衡分别需要3和4d.闪锌矿和方铅矿在ld内的浸出率分别为38.3%和34.2%,5d内zn(Ⅱ)和Pb(Ⅱ)分别还原92.0%和89.0%.使用XRD研究两种矿物的未浸出的剩余产物,闪锌矿的组成为S、ZnSO4和少量Ca3Al2O6,方铅矿的组成为S、PbSO4和少量的Pb(S,Ge)(O,OH)4和Pb8Co(Si2O7)3.

  18. Isolated Pelvic Hyperthermochemotherapeutic Perfusion -An Experimental Study on Isolating Efficacy

    Institute of Scientific and Technical Information of China (English)

    2000-01-01

    Hyperthermochemotherapeutic perfusion model through isolated pelvic vessels was developed to evaluate the leakage of hyperthermia and drugs (such as adriamycin) from the isolated pelvic circulation to systemic circulation and its associated side/toxic effects. The isolated pelvic circulation was perfused through a femoral artery catheter with hyperthermic (48 ℃ to 55 ℃) adriamycin solution (50 μg/ml) for 30 min. The efflux was drained through a femoral vein catheter. And the pelvic temperature was kept at the level of 43±0.5 ℃. The temperature of pelvic circulation was kept at 4 ℃ to 5 ℃ greater than the systemic/core temperature. The adriamycin concentration of pelvic efflux was 12 to 46 folds of that of systemic serum. The difference between them was very significant (P<0.001). As the perfusion pressure was increased, which kept lower than the mean systemic artery pressure, the leakage of the adriamycin from the isolated pelvic circulation to systemic circulation was increased, but there was no significant difference between them (P>0.05). During isolated perfusion, the systemic blood dynamics remained stable and there were no organic injuries on the important organs. It was suggested that the isolating efficacy of the modality of isolated pelvic hyperthermochemotherapeutic perfusion through vessels was rather high. The hyperthermia and drugs could be effectively limited in the isolated pelvic region with minor side effects on the systemic circulation and important organs.

  19. Geometry of isolated horizons

    CERN Document Server

    Flandera, Aleš

    2016-01-01

    While the formalism of isolated horizons is known for some time, only quite recently the near horizon solution of Einstein's equations has been found in the Bondi-like coordinates by Krishnan in 2012. In this framework, the space-time is regarded as the characteristic initial value problem with the initial data given on the horizon and another null hypersurface. It is not clear, however, what initial data reproduce the simplest physically relevant black hole solution, namely that of Kerr-Newman which describes stationary, axisymmetric black hole with charge. Moreover, Krishnan's construction employs the non-twisting null geodesic congruence and the tetrad which is parallelly propagated along this congruence. While the existence of such tetrad can be easily established in general, its explicit form can be very difficult to find and, in fact it has not been provided for the Kerr-Newman metric. The goal of this thesis was to fill this gap and provide a full description of the Kerr-Newman metric in the framework ...

  20. Isolated subtalar arthrodesis.

    Science.gov (United States)

    Yildirim, Timur; Sofu, Hakan; Çamurcu, Yalkin; Özcan, Çağri; Öner, Ali; Şahin, Vedat

    2015-03-01

    The aim of this study is to review the results of isolated subtalar arthrodesis in adults and to make a comparative analysis of the clinical outcomes between the patients with posttraumatic subtalar arthritis and the ones with other etiologic factors, and to evaluate the effects of grafting. This study included 19 men and 12 women. The mean postoperative follow-up was 36.8 months. The mean AOFAS hindfoot score improved from a mean of 46 preoperatively to a mean of 77.3 postoperatively. Thirty-one of 33 arthrodeses achieved bony union at a mean time of 15.7 weeks. The mean increase in the talocalcaneal height was 3.8 mm in the feet operated without grafting, whereas it was 8.1 mm in the feet for which grafting was performed. Improvement in talocalcaneal angle was significantly better in the feet operated with bone grafting. The feet with posttraumatic subtalar arthritis were more prone to Reflex Sympathetic Dystrophy.

  1. Seismic isolation for Advanced LIGO

    CERN Document Server

    Abbott, R; Allen, G; Cowley, S; Daw, E; Debra, D; Giaime, J; Hammond, G; Hammond, M; Hardham, C; How, J; Hua, W; Johnson, W; Lantz, B; Mason, K; Mittleman, R; Nichol, J; Richman, S; Rollins, J; Shoemaker, D; Stapfer, G; Stebbins, R

    2002-01-01

    The baseline design concept for a seismic isolation component of the proposed 'Advanced LIGO' detector upgrade has been developed with proof-of-principle experiments and computer models. It consists of a two-stage in-vacuum active isolation platform that is supported by an external hydraulic actuation stage. Construction is underway for prototype testing of a full-scale preliminary design.

  2. Improved Active Vibration Isolation Systems

    Institute of Scientific and Technical Information of China (English)

    2007-01-01

    The control force, feedback gain, and actuator stroke of several active vibration isolation systems were analyzed based on a single-layer active vibration isolation system. The analysis shows that the feedback gain and actuator stroke cannot be selected independently and the active isolation system design must make a compromise between the feedback gain and actuator stroke. The performance of active isolation systems can be improved by the joint vibration reduction using an active vibration isolation system with an adaptive dynamic vibration absorber. The results show that the joint vibration reduction method can successfully avoid the compromise between the feedback gain and actuator stroke. The control force and the object vibration amplitude are also greatly reduced.

  3. Dynamic isolation technologies in negative pressure isolation wards

    CERN Document Server

    Xu, Zhonglin

    2017-01-01

    This book presents novel design principles and technologies for dynamic isolation based on experimental studies. These approaches have now become the local standard in Beijing and are currently being promoted for use nationwide. Further, the book provides details of measures and guidelines for the design process. Departing from the traditional understanding that isolation wards should be designed with high negative pressure, airtight doors and fresh air, it establishes the basis for designing biological clean rooms, including isolation wards, using a simple and convenient scientific approach. This book is intended for designers, engineers, researchers, hospital management staff and graduate students in heating ventilation air conditioning (HVAC), air cleaning technologies and related areas.

  4. Isolated Proofs of Knowledge and Isolated Zero Knowledge

    DEFF Research Database (Denmark)

    Damgård, Ivan Bjerre; Nielsen, Jesper Buus; Wichs, Daniel

    2008-01-01

    . This is because the prover may just be forwarding messages between the environment and the verifier while the environment performs all the necessary computation. In this paper we consider an ℓ-isolated prover, which is restricted to exchanging at most ℓ bits of information with its environment. We introduce a new...... notion called ℓ-isolated proofs of knowledge (ℓ-IPoK). These protocols securely ensure that an ℓ-isolated prover knows the witness. To prevent the above-mentioned attack, an ℓ-IPoK protocol has to have communication complexity greater than ℓ. We show that for any relation in NP and any value ℓ...

  5. [Isolation, identification and oxidizing characterization of an iron-sulfur oxidizing bacterium LY01 from acid mine drainage].

    Science.gov (United States)

    Liu, Yu-jiao; Yang, Xin-ping; Wang, Shi-mei; Liang, Yin

    2013-05-01

    An acidophilic iron-sulfur oxidizing bacterium LY01 was isolated from acid mine drainage of coal in Guizhou Province, China. Strain LY01 was identified as Acidithiobacillusferrooxidans by morphological and physiological characteristics, and phylogenetic analysis of its 16S rRNA gene sequence. Strain LY01 was able to grow using ferrous ion (Fe2+), elemental sulfur (S0) and pyrite as sole energy source, respectively, but significant differences in oxidation efficiency and bacterial growth were observed when different energy source was used. When strain LY01 was cultured in 9K medium with 44.2 g x L(-1) FeSO4.7H2O as the substrate, the oxidation efficiency of Fe2+ was 100% in 30 h and the cell number of strain LY01 reached to 4.2 x 10(7) cell x mL(-1). When LY01 was cultured in 9K medium with 10 g x L(-1) S0 as the substrate, 6.7% S0 oxidation efficiency, 2001 mg x L(-1) SO4(2-) concentration and 8.9 x 10(7) cell x mL(-1) cell number were observed in 21 d respectively. When LY01 was cultured with 30 g x L(-1) pyrite as the substrate, the oxidation efficiency of pyrite, SO4(2-) concentration and cell number reached 10%, 4443 mg x L(-1) and 3.4 x 10(8) cell x mL(-1) respectively in 20 d. The effects of different heavy metals (Ni2+, Pb2+) on oxidation activity of strain LY01 cultured with pyrite were investigated. Results showed that the oxidation activity of strain LY01 was inhibited to a certain extent with the addition of Ni2+ at 10-100 mg x L(-1) to the medium, but the addition of 10-100 mg x L(-1) Pb2+ had no effect on LY01 activity.

  6. Helicopter gearbox isolation using periodically layered fluidic isolators

    Science.gov (United States)

    Szefi, Joseph Thomas

    2003-07-01

    In rotorcraft transmissions, vibration generation by meshing gear pairs is a significant source of vibration and cabin noise. This high-frequency gearbox noise is primarily transmitted to the fuselage through rigid connections, which do not appreciably attenuate vibratory energy. The high-frequency vibrations typically include discrete gear-meshing frequencies in the range of 500--2000 Hz, and are often considered irritating and can reduce pilot effectiveness and passenger comfort. Periodically-layered isolators were identified as potential passive attenuators of these high frequency vibrations. Layered isolators exhibit transmissibility "stop bands," or frequency ranges in which there is very low transmissibility. An axisymmetric model was developed to accurately predict the location of these stop bands for isolators in compression. A Ritz approximation method was used to model the axisymmetric elastic behavior of layered cylindrical isolators. This model of layered isolators was validated with experiments. The physical design constraints of the proposed helicopter gearbox isolators were then estimated. Namely, constraints associated with isolator mass, axial stiffness, geometry, and elastomeric fatigue were determined. The passive performance limits of layered isolators were then determined using a design optimization methodology employing a simulated annealing algorithm. The results suggest that layered isolators cannot always meet frequency targets given a certain set of design constraints. Many passive and active design enhancements were considered to address this problem, and the use of embedded inertial amplifiers was found to exhibit a combination of advantageous effects. The first benefit was a lowering of the beginning stop band frequency, and thus a widening of the original stop band. The second was a tuned absorber effect, where the elastomer layer stiffness and the amplified tuned mass combined to act as a vibration absorber within the stop band. The

  7. Isolation of rat adrenocortical mitochondria

    Energy Technology Data Exchange (ETDEWEB)

    Solinas, Paola [Department of Pharmacology, School of Medicine, Case Western Reserve University, Cleveland, OH 44106 (United States); Department of Medicine, Center for Mitochondrial Disease, School of Medicine, Case Western Reserve University, Cleveland, OH 44106 (United States); Fujioka, Hisashi [Electron Microscopy Facility, Department of Pharmacology, Center for Mitochondrial Disease, School of Medicine, Case Western Reserve University, Cleveland, OH 44106 (United States); Tandler, Bernard [Department of Biological Sciences, School of Dental Medicine, Center for Mitochondrial Disease, School of Medicine, Case Western Reserve University, Cleveland, OH 44106 (United States); Hoppel, Charles L., E-mail: charles.hoppel@case.edu [Department of Pharmacology, School of Medicine, Case Western Reserve University, Cleveland, OH 44106 (United States); Department of Medicine, Center for Mitochondrial Disease, School of Medicine, Case Western Reserve University, Cleveland, OH 44106 (United States)

    2012-10-12

    Highlights: Black-Right-Pointing-Pointer A method for isolation of adrenocortical mitochondria from the adrenal gland of rats is described. Black-Right-Pointing-Pointer The purified isolated mitochondria show excellent morphological integrity. Black-Right-Pointing-Pointer The properties of oxidative phosphorylation are excellent. Black-Right-Pointing-Pointer The method increases the opportunity of direct analysis of adrenal mitochondria from small animals. -- Abstract: This report describes a relatively simple and reliable method for isolating adrenocortical mitochondria from rats in good, reasonably pure yield. These organelles, which heretofore have been unobtainable in isolated form from small laboratory animals, are now readily accessible. A high degree of mitochondrial purity is shown by the electron micrographs, as well as the structural integrity of each mitochondrion. That these organelles have retained their functional integrity is shown by their high respiratory control ratios. In general, the biochemical performance of these adrenal cortical mitochondria closely mirrors that of typical hepatic or cardiac mitochondria.

  8. Isolated Echinococcosis of cervical region

    National Research Council Canada - National Science Library

    Khare, Pratima; Kala, Pooja; Gupta, Renu; Chauhan, Nidhi

    2014-01-01

    .... We report here a very rare case of isolated hydatid cyst in a 45-year-old female patient, who presented with swelling in right cervical region about 5 cm below the angle of mandible with no evidence...

  9. Atomically thin nonreciprocal optical isolation

    Science.gov (United States)

    Lin, Xiao; Wang, Zuojia; Gao, Fei; Zhang, Baile; Chen, Hongsheng

    2014-01-01

    Optical isolators will play a critical role in next-generation photonic circuits, but their on-chip integration requires miniaturization with suitable nonreciprocal photonic materials. Here, we theoretically demonstrate the thinnest possible and polarization-selective nonreciprocal isolation for circularly polarized waves by using graphene monolayer under an external magnetic field. The underlying mechanism is that graphene electron velocity can be largely different for the incident wave propagating in opposite directions at cyclotron frequency, making graphene highly conductive and reflective in one propagation direction while transparent in the opposite propagation direction under an external magnetic field. When some practical loss is introduced, nonreciprocal isolation with graphene monolayer still possesses good performance in a broad bandwidth. Our work shows the first study on the extreme limit of thickness for optical isolation and provides theoretical guidance in future practical applications. PMID:24569672

  10. Waste Isolation Pilot Plant Overview

    Energy Technology Data Exchange (ETDEWEB)

    Weaver, Douglas James [Los Alamos National Lab. (LANL), Los Alamos, NM (United States)

    2017-03-27

    The mission of Waste Isolation Pilot Plant (WIPP) is to demonstrate the safe, environmentally sound, cost effective, permanent disposal of Transuranic (TRU) waste left from production of nuclear weapons.

  11. Pneumothorax complicating isolated clavicle fracture.

    Science.gov (United States)

    Hani, Redouane; Ennaciri, Badr; Jeddi, Idriss; El Bardouni, Ahmed; Mahfoud, Mustapha; Berrada, Mohamed Saleh

    2015-01-01

    Isolated clavicle fractures are among the commonest of traumatic fractures in the emergency department. Complications of isolated clavicle fractures are rare. Pneumothorax has been described as a complication of a fractured clavicle only rarely in English literature. In all the reported cases, the pneumothorax was treated by a thoracostomy and the clavicle fracture was treated conservatively. In our case, the pneumothorax required a chest drain insertion and the clavicle fracture was treated surgically with good result.

  12. Base isolation of fluid containers

    Energy Technology Data Exchange (ETDEWEB)

    Kim, S. [Cygna Group Inc./ICF Kaiser International, Oakland, CA (United States)

    1995-12-01

    Fluid containers often constitute critical internal equipment in power plants. However, due to possible structure-equipment interaction effect they are particularly vulnerable during strong earthquake events. An effective technique for protecting fluid containers is base isolation. By deflecting the possible seismic input energy into the superstructure, base isolation can substantially reduce seismic demand on the containers, making it more cost effective than equivalent conventional design.

  13. AN INVESTIGATION OF BASE ISOLATION

    OpenAIRE

    HABASHI, Akbar Sheidaei

    2012-01-01

    The effect of weak shear rigidity is important in the analysis of elastomeric bearings used for base isolation of buildings. In particular, the buckling load is low and the P-Δ effect is significant as compared to standard columns neglecting shear deformations. In this paper, a simple mechanical model accounting for both shear and flexural deformations is proposed to treat the P-Δ effect of elastomeric isolation bearings. With adequate choice of the model parameters, the simplified model agre...

  14. Neuroimmunological Responses to Social Isolation

    OpenAIRE

    Ramirez DDS, MSc, PhD, Karol

    2015-01-01

    Objective social isolation and perceived social isolation are psychosocial stressors that may impair the normal functioning of the neuroimmune system. Chronic activation of the neuro-immuno-endocrine communication and the consequent loss of homeostasis may lead to the appearance of pathologies and associated mood disorders. For example, alterations in the hypothalamic-pituitary- adrenal axis and sympathetic nervous system dynamics may account for the observed predisposition to inflammatory di...

  15. Market study: Biological isolation garment

    Science.gov (United States)

    1975-01-01

    The biological isolation garment was originally designed for Apollo astronauts to wear upon their return to earth from the moon to avoid the possibility of their contaminating the environment. The concept has been adapted for medical use to protect certain patients from environmental contamination and the risk of infection. The nature and size of the anticipated market are examined with certain findings and conclusions relative to clinical acceptability and potential commercial viability of the biological isolation garment.

  16. Isolation, biochemical and molecular characterization of 2 ...

    African Journals Online (AJOL)

    Isolation, biochemical and molecular characterization of ... Abstract. Pure cultures of 2-chlorophenol degrading bacteria were isolated from a natural enrichment that may be adapted to ... three isolates out of six yielded positive PCR products.

  17. Progress in Exosome Isolation Techniques

    Science.gov (United States)

    Li, Pin; Kaslan, Melisa; Lee, Sze Han; Yao, Justin; Gao, Zhiqiang

    2017-01-01

    Exosomes are one type of membrane vesicles secreted into extracellular space by most types of cells. In addition to performing many biological functions particularly in cell-cell communication, cumulative evidence has suggested that several biological entities in exosomes like proteins and microRNAs are closely associated with the pathogenesis of most human malignancies and they may serve as invaluable biomarkers for disease diagnosis, prognosis, and therapy. This provides a commanding impetus and growing demands for simple, efficient, and affordable techniques to isolate exosomes. Capitalizing on the physicochemical and biochemical properties of exosomes, a number of techniques have been developed for the isolation of exosomes. This article summarizes the advances in exosome isolation techniques with an emphasis on their isolation mechanism, performance, challenges, and prospects. We hope that this article will provide an overview of exosome isolation techniques, opening up new perspectives towards the development more innovative strategies and devices for more time saving, cost effective, and efficient isolations of exosomes from a wide range of biological matrices. PMID:28255367

  18. Geomorphology of Minnesota - Isolated Landform Structures

    Data.gov (United States)

    Minnesota Department of Natural Resources — Geomorphology of Minnesota - Isolated Landform Structures are essentially cartographic arcs representing isolated glacial features that were mapped in conjunction...

  19. Base isolation system and verificational experiment of base isolated building

    Energy Technology Data Exchange (ETDEWEB)

    Takeuchi, Mikio; Harada, Osamu; Aoyagi, Sakae; Matsuda, Taiji

    1987-05-15

    With the objective of rationalization of the earthquake resistant design and the economical design based thereupon, many base isolation systems have been proposed and its research, development and application have been made in recent years. In order to disseminate the system, it is necessary to accumulate the data obtained from vibration tests and earthquake observations and verify the reliability of the system. From this viewpoint, the Central Research Institute of Electric power Industry and Okumura Corporation did the following experiments with a base isolated building as the object: 1) static power application experiments, 2) shaking experiments, 3) free vibration experiments, 4) regular slight vibration observations and 5) earthquake response observations (continuing). This article reports the outline of the base isolation system and the base isolated building concerned as well as the results of the verification experiments 1) through 3) above. From the results of these verification experiments, the basic vibration characteristics of the base isolation system consisting of laminated rubber and plastic damper were revealed and its functions were able to be verified. Especially during the free vibration experiments, the initial displacement even up to a maximum of 10cm was applied to the portion between the foundation and the structure and this displacement corresponds to the responded amplitude in case of the earthquake of seismic intensity of the 6th degree. It is planned to continue the verification further. (18 figs, 3 tabs, 3 photos, 6 refs)

  20. Entropy of Isolated Horizons revisited

    CERN Document Server

    Basu, Rudranil; Majumdar, Parthasarathi

    2009-01-01

    The decade-old formulation of the isolated horizon classically and within loop quantum gravity, and the extraction of the microcanonical entropy of such a horizon from this formulation, is reviewed, in view of recent renewed interest. There are two main approaches to this problem: one employs an SU(2) Chern-Simons theory describing the isolated horizon degrees of freedom, while the other uses a reduced U(1) Chern-Simons theory obtained from the SU(2) theory, with appropriate constraints imposed on the spectrum of boundary states `living' on the horizon. It is shown that both these ways lead to the same infinite series asymptotic in horizon area for the microcanonical entropy of an isolated horizon. The leading area term is followed by an unambiguous correction term logarithmic in area with a coefficient $-\\frac32$, with subleading corrections dropping off as inverse powers of the area.

  1. VIBRATION ISOLATION SYSTEM PROBABILITY ANALYSIS

    Directory of Open Access Journals (Sweden)

    Smirnov Vladimir Alexandrovich

    2012-10-01

    Full Text Available The article deals with the probability analysis for a vibration isolation system of high-precision equipment, which is extremely sensitive to low-frequency oscillations even of submicron amplitude. The external sources of low-frequency vibrations may include the natural city background or internal low-frequency sources inside buildings (pedestrian activity, HVAC. Taking Gauss distribution into account, the author estimates the probability of the relative displacement of the isolated mass being still lower than the vibration criteria. This problem is being solved in the three dimensional space, evolved by the system parameters, including damping and natural frequency. According to this probability distribution, the chance of exceeding the vibration criteria for a vibration isolation system is evaluated. Optimal system parameters - damping and natural frequency - are being developed, thus the possibility of exceeding vibration criteria VC-E and VC-D is assumed to be less than 0.04.

  2. Isolated Echinococcosis of cervical region

    Directory of Open Access Journals (Sweden)

    Pratima Khare

    2014-01-01

    Full Text Available Echinococcosis, commonly called as hydatid disease, is a parasitic infestation caused by the larva of the genus Echinococcus in human. Isolated occurrence of Echinococcosis without any evidence of visceral disease is very rare. A thorough search of the literature revealed only 11 cases of isolated cervical Echinococcosis. We report here a very rare case of isolated hydatid cyst in a 45-year-old female patient, who presented with swelling in right cervical region about 5 cm below the angle of mandible with no evidence of the disease elsewhere in the body. The case was diagnosed on fine needle aspiration cytology. The diagnosis was further supported by histopathology. We propose that the treating physician should also consider the differential diagnosis of Echinococcosis in the presence of an asymptomatic soft tissue mass, especially when the patient lives in an endemic area.

  3. Solo doctors and ethical isolation.

    Science.gov (United States)

    Cooper, R J

    2009-11-01

    This paper uses the case of solo doctors to explore whether working in relative isolation from one's peers may be detrimental to ethical decision-making. Drawing upon the relevance of communication and interaction for ethical decision-making in the ethical theories of Habermas, Mead and Gadamer, it is argued that doctors benefit from ethical discussion with their peers and that solo practice may make this more difficult. The paper identifies a paucity of empirical research related to solo practice and ethics but draws upon more general medical ethics research and a study that identified ethical isolation among community pharmacists to support the theoretical claims made. The paper concludes by using the literary analogy of Soderberg's Doctor Glas to illustrate the issues raised and how ethical decision-making in relative isolation may be problematical.

  4. Isolation and use of rafts.

    Science.gov (United States)

    Brown, Deborah A

    2002-11-01

    This unit describes methods for isolating and analyzing rafts by detergent insolubility. To distinguish these rafts from raft-like membranes isolated by other methods, they are referred to here as detergent-resistant membranes (DRMs). DRMs can be isolated by flotation on sucrose density gradients or by pelleting after detergent extraction. DRM proteins can be analyzed by SDS-PAGE and immunoblotting. Additionally, radiolabeled DRM proteins can be analyzed, and lipids can be quantitated by high-performance thin layer chromatography. Support protocols needed for the lipid analysis are also provided. Finally, protocols for raft disruption by cholesterol removal and measuring the kinetics of such removal are included together with a method that reverses the cholesterol removal (cholesterol repletion).

  5. Bacteria isolated from amoebae/bacteria consortium

    Science.gov (United States)

    Tyndall, Richard L.

    1995-01-01

    New protozoan derived microbial consortia and method for their isolation are provided. Consortia and bacteria isolated therefrom are useful for treating wastes such as trichloroethylene and trinitrotoluene. Consortia, bacteria isolated therefrom, and dispersants isolated therefrom are useful for dispersing hydrocarbons such as oil, creosote, wax, and grease.

  6. Active Fault Isolation in MIMO Systems

    DEFF Research Database (Denmark)

    Niemann, Hans Henrik; Poulsen, Niels Kjølstad

    2014-01-01

    Active fault isolation of parametric faults in closed-loop MIMO system s are considered in this paper. The fault isolation consists of two steps. T he first step is group- wise fault isolation. Here, a group of faults is isolated from other pos sible faults in the system. The group-wise fault iso...

  7. Completely Isolated? Health Information Seeking among Social Isolates

    Science.gov (United States)

    Askelson, Natoshia M.; Campo, Shelly; Carter, Knute D.

    2011-01-01

    To better target messages it is important to determine where people seek their health information. Interpersonal networks are a common way most people gather health information, but some people have limited networks. Using data from the 2004 General Social Survey (N = 984), we compared social isolates and nonisolates in their health…

  8. Completely Isolated? Health Information Seeking among Social Isolates

    Science.gov (United States)

    Askelson, Natoshia M.; Campo, Shelly; Carter, Knute D.

    2011-01-01

    To better target messages it is important to determine where people seek their health information. Interpersonal networks are a common way most people gather health information, but some people have limited networks. Using data from the 2004 General Social Survey (N = 984), we compared social isolates and nonisolates in their health…

  9. Is an isolated cleft lip an isolated anomaly?

    NARCIS (Netherlands)

    Deelder, J. D.; Breugem, C. C.; de Vries, Iris; de Bruin, M.; van der Molen, A. B. Mink; van der Horst, C. M. A. M.

    Introduction: It is well known that patients with cleft lip/palate or cleft palate can have associated anomalies. However, there is a relative paucity of information about the possible anomalies associated with an isolated cleft lip. A recent study (Vallino et al., 2008) showed that children with

  10. Is an isolated cleft lip an isolated anomaly?

    NARCIS (Netherlands)

    Deelder, J. D.; Breugem, C. C.; de Vries, Iris; de Bruin, M.; van der Molen, A. B. Mink; van der Horst, C. M. A. M.

    2011-01-01

    Introduction: It is well known that patients with cleft lip/palate or cleft palate can have associated anomalies. However, there is a relative paucity of information about the possible anomalies associated with an isolated cleft lip. A recent study (Vallino et al., 2008) showed that children with cl

  11. Optical Isolator Utilizing Surface Plasmons

    Directory of Open Access Journals (Sweden)

    Shinji Yuasa

    2012-05-01

    Full Text Available Feasibility of usage of surface plasmons in a new design of an integrated optical isolator has been studied. In the case of surface plasmons propagating at a boundary between a transition metal and a double-layer dielectric, there is a significant difference of optical loss for surface plasmons propagating in opposite directions. Utilizing this structure, it is feasible to fabricate a competitive plasmonic isolator, which benefits from a broad wavelength operational bandwidth and a good technological compatibility for integration into the Photonic Integrated Circuits (PIC. The linear dispersion relation was derived for plasmons propagating in a multilayer magneto-optical slab.

  12. Isolation and Cultivation of Anaerobes

    DEFF Research Database (Denmark)

    Aragao Börner, Rosa

    2016-01-01

    Anaerobic microorganisms play important roles in different biotechnological processes. Their complex metabolism and special cultivation requirements have led to less isolated representatives in comparison to their aerobic counterparts.In view of that, the isolation and cultivation of anaerobic...... microorganisms is still a promising venture, and conventional methodologies as well as considerations and modifications are presented here. An insight into new methodologies and devices as well as a discussion on future perspectives for the cultivation of anaerobes may open the prospects of the exploitation...

  13. Isolation contactor state control system

    Energy Technology Data Exchange (ETDEWEB)

    Bissontz, Jay E.

    2017-05-16

    A controller area network (CAN) installed on a hybrid electric vehicle provides one node with control of high voltage power distribution system isolation contactors and the capacity to energize a secondary electro-mechanical relay device. The output of the secondary relay provides a redundant and persistent backup signal to the output of the node. The secondary relay is relatively immune to CAN message traffic interruptions and, as a result, the high voltage isolation contactor(s) are less likely to transition open in the event that the intelligent output driver should fail.

  14. Spontaneous isolated celiac artery dissection

    Directory of Open Access Journals (Sweden)

    Tuba Cimilli Ozturk

    2011-01-01

    Full Text Available Dyspepsia with mild, stabbing epigastric discomfort without history of trauma is a very common symptom that emergency physicians see in their daily practice. Vascular emergencies, mostly the aortic dissection and aneurysm, are always described in the differential diagnosis with persistent symptoms. Isolated celiac artery dissection occurring spontaneously is a very rare diagnosis. The involvement of branch vessels is generally observed and patients show various clinical signs and symptoms according to the involved branch vessel. Here we are presenting a case with spontaneous isolated celiac artery dissection, without any branch vessel involvement or visceral damage, detected by computed tomography scans taken on admission.

  15. Isolated complete corpus callosal agenesis

    Directory of Open Access Journals (Sweden)

    Jaiganesh S, Venkateshwaran A, Naresh Kumar C, Rajasekhar KV

    2014-11-01

    Full Text Available Isolated complete corpus callosal agenesis is a rare entity. Usually this condition will be an associated finding in other syndromes. 3 month old male child came with complaints of deformed foot on both sides, not having a social smile and neck holding. Patient referred to the Radiology department for MRI brain which showed complete absence of corpus callosum, widely separated and parallely placed lateral ventricles, colpocephaly, high riding of 3rd ventricle and absence of cingulate gyrus and radial arrangement of gyri along the interhemispheric fissure. Hence it was reported as isolated complete corpus callosal agenesis and this article describes the Embryogenesis, anatomy, developmental anomalies and its clinical manifestations & prognosis.

  16. [Isolated giant hydatid in kidney].

    Science.gov (United States)

    Ozgör, Faruk; Erbin, Akif; Berberoğlu, Ahmet Yalçın; Binbay, Murat; Sarılar, Omer; Müslümanoğlu, Ahmet Yaser

    2014-06-01

    Cyst hydatid of the kidney is parasitic condition caused by Echinococcus granulosus and identified in many countries, especially associated with sheep farming. Echinococcal larvae enter the bloodstream using the digestive system and invade any organs in the human body. The urinary system is the third most common area affected by parasitic infection after liver and lungs, but isolated renal involvement is a very rare situation, even in endemic areas. İn our case, we aimed to report a 57-year-old female patient with an 18-centimeter isolated renal cyst hydatid treated by retroperitoneal nephrectomy. The diagnosis was based on imaging findings and confirmed by histopathologically.

  17. Isolation of Leptospira from blood culture bottles.

    Science.gov (United States)

    Girault, Dominique; Soupé-Gilbert, Marie-Estelle; Geroult, Sophie; Colot, Julien; Goarant, Cyrille

    2017-01-31

    With the increasing use of real-time PCR techniques, Leptospira isolation has mostly been abandoned for the diagnosis of human leptospirosis. However, there is a great value of collecting Leptospira isolates to better understand the epidemiology of this complex zoonosis and to provide the researchers with different isolates. In this study, we have successfully isolated different Leptospira strains from BacT/Alert aerobic blood culture bottles and suggest that this privileged biological material offers an opportunity to isolate leptospires.

  18. Innovative Bioreactor Development for Methanotrophic Biodegradation of Trichloroethylene

    Science.gov (United States)

    1994-01-01

    in this portion of the reator. Table 8 shows model generated data of TCE man flux rates, TCE concentrations, and biomasa concentrations throughout...of the model on biomass with the three TCE feed concentrations examined (Table 8). The model was also examined for CSTR. biomasa concentrations of 1809

  19. Effectiveness of Base Isolation Technique and Influence of Isolator Characteristics on Response of a Base Isolated Building

    Directory of Open Access Journals (Sweden)

    Sunita Tolani

    2016-06-01

    Full Text Available This study concerns with the seismic response comparison of a fixed base building with a base isolated building and parametric study of a base isolated building. The structural system considered for analysis is a three storey reinforced concrete building, which is idealized as a shear type building with one lateral degree of freedom at each floor level. The isolation systems considered for this study are Laminated Rubber bearing (LRB, Lead Rubber Bearing (N-Z bearing and Friction Pendulum System (FPS. The response of fixed base building and of base isolated building is compared in terms of maximum top floor acceleration, interstorey drift, maximum floor displacements and base shear. For parametric study important isolation system parameters considered are: (i isolation time period, isolator damping for LRB; (ii isolator yield strength, isolation time period, isolator damping for N-Z bearing and (iii isolation time period, friction coefficient for FPS. It is found that base isolation technique is very effective in reducing seismic response of structure and isolation system parameters significantly influence the earthquake response of a base isolated structure.

  20. Providencia stuartii Isolates from Greece

    NARCIS (Netherlands)

    Oikonomou, Olga; Liakopoulos, Apostolos; Phee, Lynette M.; Betts, Jonathan; Mevius, Dik; Wareham, David W.

    2016-01-01

    Providencia stuartii has emerged as an important nosocomial pathogen. We describe an outbreak due to a multidrug-resistant strain over a 4-month period in a critical care unit in Athens. Molecular typing revealed each of the isolates to be clonally related with coresistance to cephalosporins, car

  1. IMPROVED ISOLATION & ELECTROSPINNING OF ZEIN

    Science.gov (United States)

    The major historical use of zein, the predominant corn protein, has been in the fibers market. In order to evaluate new formulations of zein in the fibers market a simple means of isolating and generating fibers is necessary. We have evaluated the ability to electrospin zein from numerous solvents...

  2. Isolated superior mesenteric artery dissection

    Directory of Open Access Journals (Sweden)

    Lalitha Palle

    2010-01-01

    Full Text Available Isolated superior mesenteric artery (SMA dissection without involvement of the aorta and the SMA origin is unusual. We present a case of an elderly gentleman who had chronic abdominal pain, worse after meals. CT angiography, performed on a 64-slice CT scanner, revealed SMA dissection with a thrombus. A large artery of Drummond was also seen. The patient was managed conservatively.

  3. Mitochondria: isolation, structure and function.

    Science.gov (United States)

    Picard, Martin; Taivassalo, Tanja; Gouspillou, Gilles; Hepple, Russell T

    2011-09-15

    Mitochondria are complex organelles constantly undergoing processes of fusion and fission, processes that not only modulate their morphology, but also their function. Yet the assessment of mitochondrial function in skeletal muscle often involves mechanical isolation of the mitochondria, a process which disrupts their normally heterogeneous branching structure and yields relatively homogeneous spherical organelles. Alternatively, methods have been used where the sarcolemma is permeabilized and mitochondrial morphology is preserved, but both methods face the downside that they remove potential influences of the intracellular milieu on mitochondrial function. Importantly, recent evidence shows that the fragmented mitochondrial morphology resulting from routine mitochondrial isolation procedures used with skeletal muscle alters key indices of function in a manner qualitatively similar to mitochondria undergoing fission in vivo. Although these results warrant caution when interpreting data obtained with mitochondria isolated from skeletal muscle, they also suggest that isolated mitochondrial preparations might present a useful way of interrogating the stress resistance of mitochondria. More importantly, these new findings underscore the empirical value of studying mitochondrial function in minimally disruptive experimental preparations. In this review, we briefly discuss several considerations and hypotheses emerging from this work.

  4. Isolation by distance, web service

    Directory of Open Access Journals (Sweden)

    Bohonak Andrew J

    2005-03-01

    Full Text Available Abstract Background The population genetic pattern known as "isolation by distance" results from spatially limited gene flow and is a commonly observed phenomenon in natural populations. However, few software programs exist for estimating the degree of isolation by distance among populations, and they tend not to be user-friendly. Results We have created Isolation by Distance Web Service (IBDWS a user-friendly web interface for determining patterns of isolation by distance. Using this site, population geneticists can perform a variety of powerful statistical tests including Mantel tests, Reduced Major Axis (RMA regression analysis, as well as calculate FST between all pairs of populations and perform basic summary statistics (e.g., heterozygosity. All statistical results, including publication-quality scatter plots in Postscript format, are returned rapidly to the user and can be easily downloaded. Conclusion IBDWS population genetics analysis software is hosted at http://phage.sdsu.edu/~jensen/ and documentation is available at http://www.bio.sdsu.edu/pub/andy/IBD.html. The source code has been made available on Source Forge at http://sourceforge.net/projects/ibdws/.

  5. Vapor-barrier Vacuum Isolation System

    Science.gov (United States)

    Weinstein, Leonard M. (Inventor); Taminger, Karen M. (Inventor)

    2014-01-01

    A system includes a collimated beam source within a vacuum chamber, a condensable barrier gas, cooling material, a pump, and isolation chambers cooled by the cooling material to condense the barrier gas. Pressure levels of each isolation chamber are substantially greater than in the vacuum chamber. Coaxially-aligned orifices connect a working chamber, the isolation chambers, and the vacuum chamber. The pump evacuates uncondensed barrier gas. The barrier gas blocks entry of atmospheric vapor from the working chamber into the isolation chambers, and undergoes supersonic flow expansion upon entering each isolation chamber. A method includes connecting the isolation chambers to the vacuum chamber, directing vapor to a boundary with the working chamber, and supersonically expanding the vapor as it enters the isolation chambers via the orifices. The vapor condenses in each isolation chamber using the cooling material, and uncondensed vapor is pumped out of the isolation chambers via the pump.

  6. NASDA's activities on vibration isolation technology

    Science.gov (United States)

    1992-01-01

    The National Space Development Agency's (NASDA) activities in providing various vibration isolation technologies for the Space Station Mission are covered in viewgraph form. Technologies covered include an active vibration isolation system for extra sensitive missions in the low frequency range, a passive damping system consisting of a damping rack for the reduction of resonance amplification, and an isolator for vibration isolation from low frequencies. Information is given in viewgraph form on the active vibration isolation concept, voice coil type electromagnetic suspension, a profile of an active vibration isolation system, a three degree of freedom ground experiment, and acceleration feedback.

  7. Sympathicotomy for isolated facial blushing

    DEFF Research Database (Denmark)

    Licht, Peter Bjørn; Pilegaard, Hans K; Ladegaard, Lars

    2012-01-01

    Background. Facial blushing is one of the most peculiar of human expressions. The pathophysiology is unclear, and the prevalence is unknown. Thoracoscopic sympathectomy may cure the symptom and is increasingly used in patients with isolated facial blushing. The evidence base for the optimal level...... of targeting the sympathetic chain is limited to retrospective case studies. We present a randomized clinical trial. Methods. 100 patients were randomized (web-based, single-blinded) to rib-oriented (R2 or R2-R3) sympathicotomy for isolated facial blushing at two university hospitals during a 6-year period...... in all social and mental domains in both groups. Overall, 85% of the patients had an excellent or satisfactory result, with no significant difference between the R2 procedure and the R2-R3 procedure. Mild recurrence of facial blushing occurred in 30% of patients within the first year. One patient...

  8. Isolated urachal malakoplakia mimicking malignancy

    Directory of Open Access Journals (Sweden)

    Saisriharsha Pakalapati

    2017-01-01

    Full Text Available Malakoplakia is an unusual inflammatory disease with uncertain pathogenesis affecting any organ in the body, but predominantly genitourinary tract, with specific predilection to the bladder. We report a rare case of isolated malakoplakia of the urachus in a 29-year-old male patient who presented with lower urinary tract symptoms without any hematuria. Investigations revealed sterile pyuria with no bacterial growth in urine. Radiological investigations revealed a mass in the urachal region. The patient underwent cystoscopy with biopsy followed by pelvic lymph node dissection and partial cystectomy with excision of the urachal mass. Histopathological examination of the mass revealed malakoplakia. Postoperative course was uneventful. To the best of our knowledge, this is the first ever case report of isolated urachal malakoplakia without any concomitant malignancy or bladder involvement reported in our country and one of the very few reported worldwide.

  9. Pediatric isolated bilateral iliac aneurysm.

    Science.gov (United States)

    Chithra, R; Sundar, R Ajai; Velladuraichi, B; Sritharan, N; Amalorpavanathan, J; Vidyasagaran, T

    2013-07-01

    Aneurysms are rare in children. Isolated iliac artery aneurysms are very rare, especially bilateral aneurysms. Pediatric aneurysms are usually secondary to connective tissue disorders, arteritis, or mycotic causes. We present a case of a 3-year-old child with bilateral idiopathic common iliac aneurysms that were successfully repaired with autogenous vein grafts. Copyright © 2013 Society for Vascular Surgery. Published by Mosby, Inc. All rights reserved.

  10. Isolated complete corpus callosal agenesis

    OpenAIRE

    Jaiganesh S, Venkateshwaran A, Naresh Kumar C, Rajasekhar KV

    2014-01-01

    Isolated complete corpus callosal agenesis is a rare entity. Usually this condition will be an associated finding in other syndromes. 3 month old male child came with complaints of deformed foot on both sides, not having a social smile and neck holding. Patient referred to the Radiology department for MRI brain which showed complete absence of corpus callosum, widely separated and parallely placed lateral ventricles, colpocephaly, high riding of 3rd ventricle and absence of cingulate gyrus an...

  11. Transformerless dc-Isolated Converter

    Science.gov (United States)

    Rippel, Wally E.

    1987-01-01

    Efficient voltage converter employs capacitive instead of transformer coupling to provide dc isolation. Offers buck/boost operation, minimal filtering, and low parts count, with possible application in photovoltaic power inverters, power supplies and battery charges. In photovoltaic inverter circuit with transformerless converter, Q2, Q3, Q4, and Q5 form line-commutated inverter. Switching losses and stresses nil because switching performed when current is zero.

  12. Isolation of intraflagellar transport trains.

    Science.gov (United States)

    Mencarelli, Caterina; Mitchell, Aaron; Leoncini, Roberto; Rosenbaum, Joel; Lupetti, Pietro

    2013-08-01

    The intraflagellar transport (IFT) system was first identified in situ by electron microscopy in thin sections of plastic-embedded flagella as linear arrays of electrondense particles, located between the B tubules of the outer doublets and the flagellar membrane. These arrays of particles are referred to as IFT trains. Upon membrane rupture, IFT trains are thought to easily dissociate to yield soluble IFT particles, which are commonly purified through sucrose gradients as ∼16-17S complexes. The latters easily dissociate into two subcomplexes, named A and B. We report here the isolation, visualization, and identification by immunolabeling of flexible strings of IFT particles, which are structurally similar to in situ IFT trains and appear to be formed by both complex A and complex B polypeptides. Moreover, the particles forming isolated IFT trains are structurally similar to the individual particles found in the ∼17S gradient peak. Our results provide the first direct evidence that ∼17S particles do indeed compose the IFT trains. The paper also represents the first isolation of the IFT trains, and opens new possibilities for higher resolution studies on their structure and how particles are attached to each other to form the particle trains.

  13. Antibiosis and dark-pigments secretion by the phytopathogenic and environmental fungal species after interaction in vitro with a Bacillus subtilis isolate

    Directory of Open Access Journals (Sweden)

    Alexandre Paulo Machado

    2010-10-01

    Full Text Available In this work, different reactions in vitro between an environmental bacterial isolate and fungal species were related. The Gram-positive bacteria had terminal and subterminal endospores, presented metabolic characteristics of mesophilic and acidophilic growth, halotolerance, positive to nitrate reduction and enzyme production, as caseinase and catalase. The analysis of partial sequences containing 400 to 700 bases of the 16S ribosomal RNA gene showed identity with the genus Bacillus. However, its identity as B. subtilis was confirmed after analyses of the rpoB, gyrA, and 16S rRNA near-full-length sequences. Strong inhibitory activity of environmental microorganisms, such as Penicillium sp, Aspergillus flavus, A. niger, and phytopathogens, such as Colletotrichum sp, Alternaria alternata, Fusarium solani and F. oxysporum f.sp vasinfectum, was shown on co-cultures with B. subtilis strain, particularly on Sabouraud dextrose agar (SDA and DNase media. Red and red-ochre color pigments, probably phaeomelanins, were secreted by A. alternata and A. niger respectively after seven days of co-culture.Na presente investigação, nosso objetivo principal foi relatar diferentes interações in vitro de um isolado bacteriano ambiental com espécies fúngicas. Através da identificação clássica, nós verificamos que o bacilo ambiental apresentava endósporos terminais e subterminais, características metabólicas de mesofilia, acidofilia, halotolerância, redução de nitrato e produção de enzimas, como caseinase e catalase. Análise de seqüências parciais do gene 16S RNAr contendo de 400 a 700 bases revelou identidade com gênero Bacillus. No entanto, a espécie Bacillus subtilis foi confirmada somente depois da análise de seqüências dos genes rpoB, gyrA, and 16S RNAr. Intensa atividade inibitória aos fungos ambientais, como Penicillium sp, Aspergillus flavus, A. niger, e fitopatogênicos, como Colletotrichum sp, Alternaria alternata, Fusarium solani

  14. Isolation of Cellulolytic Actinomycetes from Marine Sediments

    OpenAIRE

    Veiga, Manuel; Esparis, Azucena; Fabregas, Jaime

    1983-01-01

    The cellulolytic activity of 36 actinomycetes strains isolated from marine sediments was investigated by the cellulose-azure method. Approximately 50% of the isolates exhibited various degrees of cellulolytic activity.

  15. Tiamulin resistance in porcine Brachyspira pilosicoli isolates.

    Science.gov (United States)

    Pringle, M; Landén, A; Franklin, A

    2006-02-01

    There are few studies on antimicrobial susceptibility of Brachyspira pilosicoli, therefore this study was performed to investigate the situation among isolates from pigs. The tiamulin and tylosin susceptibility was determined by broth dilution for 93 and 86 porcine B. pilosicoli isolates, respectively. The isolates came from clinical samples taken in Swedish pig herds during the years 2002 and 2003. The tylosin minimal inhibitory concentration (MIC) was >16 microg/ml for 50% (n=43) of the isolates tested. A tiamulin MIC >2 microg/ml was obtained for 14% (n=13) of the isolates and these were also tested against doxycycline, salinomycin, valnemulin, lincomycin and aivlosin. For these isolates the susceptibility to salinomycin and doxycycline was high but the MICs for aivlosin varied. The relationship between the 13 tiamulin resistant isolates was analyzed by pulsed-field gel electrophoresis (PFGE). Among the 13 isolates 10 different PFGE patterns were identified.

  16. Isolation, production, purification, assay and characterization of ...

    African Journals Online (AJOL)

    Isolation, production, purification, assay and characterization of fibrinolytic ... are isolated from Bacillus subtilis, β-haemolytic Streptococci and urine sample. ... recombinant E.coli containing short fragment genomic DNA of Pseudomonas sp.

  17. Mycoplasma contamination of Chlamydia pneumoniae isolates

    DEFF Research Database (Denmark)

    Huniche, BS; Jensen, Lise Torp; Birkelund, Svend

    1998-01-01

    We examined 6 C. pneumonia isolates from The American Type Culture Collection (ATCC) and 2 Finnish isolates for Mycoplasma contamination. Three of the ATCC isolates and both of the Finnish isolates were Mycoplasma-contaminated. The contaminants were characterized by means of growth in BEa and BEg...... media, immunoblotting, polymerase chain reaction and pulsed field gel electrophoresis. Two of the 6 ATCC isolates [ATCC VR1355 (TWAR strain 2043) and ATCC VR1356 (TWAR strain 2023)] were infected with Mycoplasma hominis and 1 isolate [ATCC VR2282 (TWAR strain TW183)] was contaminated with both...... Mycoplasma hominis and Mycoplasma orale, whereas 3 of the ATCC isolates [ATCC VR1310, ATCC VR1360 (TWAR strain CM-1) and ATCC 53592 (TWAR strain AR39)] were not contaminated. The Finnish C. pneumoniae isolates Kajaani 6 and Parola were found to be contaminated with M. hominis and M. orale, respectively...

  18. Isolation of haloorganic groundwater humic substances

    DEFF Research Database (Denmark)

    Krog, M.; Grøn, C.

    1995-01-01

    Humic substances were isolated from groundwater according to a revised method designed to avoid organohalogen artefacts. The prepared humic substances exhibited lower halogen contents than humic substances isolated according to the conventionally used method. Excessive oxidation or hydrolysis...

  19. Genetics Home Reference: isolated lissencephaly sequence

    Science.gov (United States)

    ... Facebook Share on Twitter Your Guide to Understanding Genetic Conditions Search MENU Toggle navigation Home Page Search ... Conditions Genes Chromosomes & mtDNA Resources Help Me Understand Genetics Home Health Conditions isolated lissencephaly sequence isolated lissencephaly ...

  20. Isolation, characterization and antimicrobial activity of Streptomyces ...

    African Journals Online (AJOL)

    DR TONUKARI

    2013-12-18

    Dec 18, 2013 ... series on all media. The results showed that glycerol yeast extract and starch casein were the best .... MATERIALS AND METHODS. Collection of .... Genomic DNA was isolated from the isolates using a bacterial genomic DNA ...

  1. Are Isolated Indigenous Populations Headed toward Extinction?

    National Research Council Canada - National Science Library

    Walker, Robert S; Kesler, Dylan C; Hill, Kim R

    2016-01-01

    .... Except in emergency situations, the current policy of governments in Brazil, Colombia, and Peru towards isolated tribes is a "leave them alone" strategy, in which isolated groups are left uncontacted...

  2. Isolated Malignant Melanoma Metastasis to the Pancreas

    Directory of Open Access Journals (Sweden)

    Anne K. Larsen, MD

    2013-11-01

    Full Text Available Summary: Malignant melanomas rarely develop isolated pancreatic metastases. We describe a unique patient who is still alive 22 years following an isolated pancreatic melanoma metastasis, and we review the sparse literature in the field.

  3. common bacterial isolates from infected eyes abstract

    African Journals Online (AJOL)

    LIVINGSTON

    The common bacterial isolates and their antibiotics susceptibility were studied in 298 bacterial ... Bacteria were isolated most on the eye infections of the conjunctiva ... practitioners to avoid development of resistance from indiscriminate use.

  4. Isolated malignant melanoma metastasis to the pancreas

    DEFF Research Database (Denmark)

    Larsen, Anne K; Krag, Christen; Geertsen, Poul

    2013-01-01

    SUMMARY: Malignant melanomas rarely develop isolated pancreatic metastases. We describe a unique patient who is still alive 22 years following an isolated pancreatic melanoma metastasis, and we review the sparse literature in the field.......SUMMARY: Malignant melanomas rarely develop isolated pancreatic metastases. We describe a unique patient who is still alive 22 years following an isolated pancreatic melanoma metastasis, and we review the sparse literature in the field....

  5. Molecular characterization of Danish Cryptosporidium parvum isolates

    DEFF Research Database (Denmark)

    Enemark, Heidi L.; Ahrens, Peter; Juel, Cynthia Dawn

    2002-01-01

    was characterized as C. meleagridis. The porcine Cryptosporidium isolates (N = 4) revealed a pattern which was genetically distinct from human and bovine isolates. Cryptosporidium in a hedgehog (Erinaceus europaeus L.) was identified for the first time. By microsatellite sequencing the hedgehog isolate showed...

  6. Fungi isolated in school buildings

    Directory of Open Access Journals (Sweden)

    Elżbieta Ejdys

    2013-12-01

    Full Text Available The aim of the study was to determine the species composition of fungi occurring on wall surfaces and in the air in school buildings. Fungi isolated from the air using the sedimentation method and from the walls using the surface swab technique constituted the study material. Types of finish materials on wall surfaces were identified and used in the analysis. Samples were collected in selected areas in two schools: classrooms, corridors, men's toilets and women's toilets, cloakrooms, sports changing rooms and shower. Examinations were conducted in May 2005 after the heating season was over. Fungi were incubated on Czapek-Dox medium at three parallel temperatures: 25, 37 and 40°C, for at least three weeks. A total of 379 isolates of fungi belonging to 32 genera of moulds, yeasts and yeast-like fungi were obtained from 321 samples in the school environment. The following genera were isolated most frequently: Aspergillus, Penicillium and Cladosporium. Of the 72 determined species, Cladosporium herbarum, Aspergillus fumigatus and Penicillium chrysogenum occurred most frequently in the school buildings. Wall surfaces were characterised by an increased prevalence of mycobiota in comparison with the air in the buildings, with a slightly greater species diversity. A certain species specificity for rough and smooth wall surfaces was demonstrated. Fungi of the genera Cladosporium and Emericella with large spores adhered better to smooth surfaces while those of the genus Aspergillus with smaller conidia adhered better to rough surfaces. The application of three incubation temperatures helped provide a fuller picture of the mycobiota in the school environment.

  7. Lipid biomarker production and preservation in acidic ecosystems: Relevance to early Earth and Mars

    Science.gov (United States)

    Jahnke, L. L.; Parenteau, M. N.; Harris, R.; Bristow, T.; Farmer, J. D.; Des Marais, D. J.

    2013-12-01

    Compared to relatively benign carbonate buffered marine environments, terrestrial Archean and Paleoproterozoic life was forced to cope with a broader range of pH values. In particular, acidic terrestrial ecosystems arose from the oxidation of reduced species in hydrothermal settings and crustal reservoirs of metal sulfides, creating acid sulfate conditions. While oxidation of reduced species is facilitated by reactions with molecular oxygen, acidic conditions also arose in Archean hydrothermal systems before the rise of oxygen (Van Kranendonk, 2006), expanding the range of time over which acidophiles could have existed on the early Earth. Acidic terrestrial habitats would have included acidic hydrothermal springs, acid sulfate soils, and possibly lakes and streams lacking substantial buffering capacity with sources of acidity in their catchments. Although acidic hot springs are considered extreme environments on Earth, robust and diverse microbial communities thrive in these habitats. Such acidophiles are found across all three domains of life and include both phototrophic and chemotrophic members. In this presentation, we examine hopanes and sterols that are characteristic of microbial communities living in acidic hydrothermal environments. Moreover we discuss taphonomic processes governing the capture and preservation of these biosignatures in acid environments. In particular, we discuss the production and early preservation of hopanoids and sterols in the following geological/mineralogical settings: 1) rapid entombment of microbes and organic matter by predominantly fine-grained silica; 2) rapid burial of organic matter by clay-rich, silica poor sediments; 3) and the survival of organics in iron oxide and sulfate rich sediments. We discovered and isolated an acid-tolerant purple non-sulfur anoxygenic phototroph from Lassen Volcanic National Park that synthesizes 3methyl-bacteriohopanepolyols. These compounds were previously thought to be exclusively made by

  8. Thermodynamic laws in isolated systems.

    Science.gov (United States)

    Hilbert, Stefan; Hänggi, Peter; Dunkel, Jörn

    2014-12-01

    The recent experimental realization of exotic matter states in isolated quantum systems and the ensuing controversy about the existence of negative absolute temperatures demand a careful analysis of the conceptual foundations underlying microcanonical thermostatistics. Here we provide a detailed comparison of the most commonly considered microcanonical entropy definitions, focusing specifically on whether they satisfy or violate the zeroth, first, and second laws of thermodynamics. Our analysis shows that, for a broad class of systems that includes all standard classical Hamiltonian systems, only the Gibbs volume entropy fulfills all three laws simultaneously. To avoid ambiguities, the discussion is restricted to exact results and analytically tractable examples.

  9. [Neuroanatomy of Isolated Body Lateropulsion].

    Science.gov (United States)

    Nakazato, Yoshihiko; Tamura, Naotoshi; Ikeda, Kei; Tanaka, Ai; Yamamoto, Toshimasa

    2016-03-01

    Axial body lateropulsion, a phenomenon where the body is pulled toward the side of the lesion, with tendency of falling down, is the well-known transient feature of lateral medullary syndrome. In some cases, axial body lateropulsion occurs without vestibular and cerebellar symptoms (isolated body lateropulsion:[iBL]). Patients with iBL have a lesion located in the spinocerebellar tract, descending lateral vestibulospinal tract, vestibulo-thalamic pathway, dentatorubrothalamic pathway, or thalamocortical fascicle. This review deals with the anatomic basis and clinical significance of iBL.

  10. Isoflavonoids isolated from Cuban propolis.

    Science.gov (United States)

    Piccinelli, Anna Lisa; Campo Fernandez, Mercedes; Cuesta-Rubio, Osmany; Márquez Hernández, Ingrid; De Simone, Francesco; Rastrelli, Luca

    2005-11-16

    Chemical investigation of a red-type Cuban propolis sample has led to the isolation of 11 isoflavonoids (2 isoflavones, 3 isoflavans, and 6 pterocarpans), together with gallic acid, isoliquiritigenin, and (-)-liquiritigenin. Structural determination, including the absolute stereochemistry, was accomplished by spectroscopic analysis, particularly CD and 2D NMR techniques. The fragmentation behavior of pterocarpans was studied by electrospray ionization (ESI) tandem mass spectrometry (MS/MS) using an ion-trap analyzer, and a generalized fragmentation pathway, useful in the identification and structural characterization of pterocarpans, is proposed. Isoflavonoids are reported for the first time from propolis samples.

  11. Simplified design procedure for base isolation system

    Energy Technology Data Exchange (ETDEWEB)

    Takayama, Mineo; Tada, Hideyuki [Fukuoka Univ. (Japan). Faculty of Engineering

    1995-12-01

    This paper presents a simplified design procedure which incorporates an existing response prediction method for base isolated buildings into design methods for isolation devices (rubber bearings and hysteresis dampers). The procedure enables a designer to easily identify the relationship between tile seismic behavior of base-isolated buildings and the characteristics of isolation devices. The prediction method, proposed by Prof. Akiyama, is based on energy balance between the total input seismic energy and the energy absorbed by the isolation devices. The method is very accurate. The design methods for devices were developed by authors based on experimental and finite element analysis results.

  12. From rumors to genetic isolates

    Directory of Open Access Journals (Sweden)

    Eduardo E. Castilla

    2014-01-01

    Full Text Available Here we propose a registration process for population genetic isolates, usually geographic clusters of genetic disorders, based on the systematic search of rumors, defined as any type of account regardless of its reliability. Systematically ascertained rumors are recorded, and validated through a progressive process of pre-established steps. This paper outlines the conceptual basis for this approach and presents the preliminary results from a rumor-based nationwide registry of genetically isolated populations, named CENISO (Censo Nacional de Isolados, operating in Brazil since 2009. During the first four years of its existence (2009-2013, a total of 191 Rumors were registered and validated, resulting in a prevalence rate of one per million inhabitants of Brazil. When the five statutory geographic regions of Brazil were considered, more Rumors were registered for the Northeast (2.11; 1.74-2.54 per 10(6 than for the remaining four regions, North, Center-West, Southeast, and South, which did not differ among themselves. About half (86/191 of the recorded rumors were proven to be geographic clusters; of these disorders, 58 were autosomal recessive, 17 autosomal dominant, 5 X-linked, 3 multifactorial, and one environmental (thalidomide embryopathy.

  13. Blackbody Radiation from Isolated Neptunes

    CERN Document Server

    Ginzburg, Sivan; Loeb, Abraham

    2016-01-01

    Recent analyses of the orbits of some Kuiper Belt objects hypothesize the presence of an undiscovered Neptune-size planet at a very large separation from the Sun. The energy budget of Neptunes on such distant orbits is dominated by the internal heat released by their cooling rather than solar irradiation (making them effectively "isolated"). The blackbody radiation that these planets emit as they cool may provide the means for their detection. Here we use an analytical toy model to study the cooling and radiation of isolated Neptunes. This model can translate a detection (or a null detection) to a constraint on the size and composition of the hypothesized "Planet Nine". Specifically, the thick gas atmosphere of Neptune-like planets serves as an insulating blanket which slows down their cooling. Therefore, a measurement of the blackbody temperature, $T_{\\rm eff}\\sim 50\\textrm{K}$, at which a Neptune emits can be used to estimate the mass of its atmosphere, $M_{\\rm atm}$. Explicitly, we find the relation $T_{\\r...

  14. Isolation of technogenic magnetic particles

    Energy Technology Data Exchange (ETDEWEB)

    Catinon, Mickaël, E-mail: mickael.catinon@gmail.com [Laboratoire LECA, UMR 5553, Equipe Pollution, Environnement, Ecotoxicologie et Ecoremédiation, Univ. J. Fourier, 38041 Grenoble (France); Ayrault, Sophie, E-mail: sophie.ayrault@lsce.ispl.fr [Laboratoire des Sciences du Climat et de l' Environnement, UMR 8212, CEA-CNRS-UVSQ/IPSL, 91198 Gif-sur-Yvette (France); Boudouma, Omar, E-mail: boudouma@ccr.jussieu.fr [Service du MEB, UFR928, Université Pierre et Marie Curie, 75252 Paris VI (France); Bordier, Louise, E-mail: Louise.Bordier@lsce.ipsl.fr [Laboratoire des Sciences du Climat et de l' Environnement, UMR 8212, CEA-CNRS-UVSQ/IPSL, 91198 Gif-sur-Yvette (France); Agnello, Gregory, E-mail: contact@evinrude.fr [Evinrude, Espace St Germain, 38200 Vienne (France); Reynaud, Stéphane, E-mail: stephane.reynaud@ujf-grenoble.fr [Laboratoire LECA, UMR 5553, Equipe Pollution, Environnement, Ecotoxicologie et Ecoremédiation, Univ. J. Fourier, 38041 Grenoble (France); Tissut, Michel, E-mail: michel.tissut@ujf-grenoble.fr [Laboratoire LECA, UMR 5553, Equipe Pollution, Environnement, Ecotoxicologie et Ecoremédiation, Univ. J. Fourier, 38041 Grenoble (France)

    2014-03-01

    Technogenic magnetic particles (TMPs) emitted by various industrial sources, such as smelting plants, end up after atmospheric transfer on the soil surface. In the present study, we characterised the origin and composition of such particles emitted by a large iron smelting plant and deposited on particular substrates, namely tombstones, which act as a very interesting and appropriate matrix when compared to soil, tree bark, lichens or attic dust. The isolation and subsequent description of TMPs require a critical step of separation between different components of the sample and the magnetic particles; here, we described an efficient protocol that fulfils such a requirement: it resorts to water suspension, sonication, repeated magnetic extraction, sedimentation, sieving and organic matter destruction at 550 °C in some instances. The isolated TMPs displayed a noticeable crystalline shape with variable compositions: a) pure iron oxides, b) iron + Cr, Ni or Zn, and c) a complex structure containing Ca, Si, Mg, and Mn. Using Scanning Electron Microscope Energy Dispersive X-ray (SEM–EDX), we obtained profiles of various and distinct magnetic particles, which allowed us to identify the source of the TMPs. - Highlights: • The developed method offers a low-cost approach of large-scale dry deposition. • Tombstones are excellent supports for sampling these atmospheric deposits. • Smelted elements crystallise after cooling, giving typical technogenic magnetic particles (TMPs). • Coupling microscopic and bulk analyses allows identifying TMP origin. • Magnetic TMPs issued from steel industry were separated by a new technique.

  15. Braking Index of Isolated Pulsars

    CERN Document Server

    Hamil, Oliver Q; Urbanec, Martin; Urbancova, Gabriela

    2016-01-01

    Isolated pulsars are rotating neutron stars with accurately measured angular velocities $\\Omega$, and their time derivatives that show unambiguously that the pulsars are slowing down. The commonly accepted view is that it arises through emission of magnetic dipole radiation (MDR) from a rotating magnetized body. The calculated energy loss by a rotating pulsar with a constant moment of inertia is assumed proportional to a model dependent power of $\\Omega$. This relation leads to the power law $\\dot{\\Omega}$ = -K $\\Omega^{\\rm n}$ where $n$ is called the braking index. The MDR model predicts $n$ exactly equal to 3. Selected observations of isolated pulsars provide rather precise values of $n$, individually accurate to a few percent or better, in the range 1$ <$ n $ < $ 2.8, which is consistently less than the predictions of the MDR model. In spite of an extensive investigation of various modifications of the MDR model, no satisfactory explanation of observation has been found yet. The aim of this work is t...

  16. Molecular characterization of bacteria isolated from the Kingdom of ...

    African Journals Online (AJOL)

    ola

    2012-11-06

    Nov 6, 2012 ... ... bacterial isolation. Bacterial isolates were evaluated for their antimicrobial activities against pathogenic fungi which cause ... Molecular techniques based on genomic DNA have ... Isolation of total DNA from isolated strains.

  17. Reovirus, isolated from SARS patients

    Institute of Scientific and Technical Information of China (English)

    DUAN Qing; SONG Lihua; GAN Yonghua; TAN Hua; JIN Baofeng; LI Huiyan; ZUO Tingting; CHEN Dehui; ZHANG Xuemin; ZHU Hong; YANG Yi; LI Weihua; ZHOU Yusen; HE Jun; HE Kun; ZHANG Haojie; ZHOU Tao

    2003-01-01

    Beijing has been severely affected by SARS, and SARS-associated coronavirus has been confirmed as its cause. However, clinical and experimental evidence implicates the possibility of co-infection. In this report, reovirus was isolated from throat swabs of SARS patients, including the first case in Beijing andher mother. Identification with the electron microscopy revealed the characteristic features of reovirus. 24 of 38 samples from other SARS cases were found to have serologic responses to the reovirus. Primers designed for reovirus have amplified several fragments of DNA, one of which was sequenced (S2 gene fragment), which indicates it as a unique reovirus (orthoreovirus). Preliminary animal experiment showed that inoculation of the reovirus in mice caused death with atypical pneumonia. Nevertheless, the association of reovirus with SARS outbreak requires to be further investigated.

  18. Multiple isolated cutaneous plexiform schwannomas

    Directory of Open Access Journals (Sweden)

    Enas A. S. Attia

    2011-01-01

    Full Text Available Plexiform schwannoma is a rare neurogenic tumor, arising from skin and subcutaneous tissue. The presence of multiple schwannomas suggests a possible association with neurofibromatosis type 2 (NF2. A 50-year old male patient presented with multiple papulo-nodular cutaneous lesions on both arms and forearms. Histopathological examination revealed a dermal multinodular pattern of well-circumscribed masses of closely packed cells, with peripheral myxoid tissue, well-encapsulated in a thin collagenous capsule. S-100 immunohistochemical staining was diffusely and strongly positive. Neuron-specific enolase was positive, confirming a neural tissue tumor. An audiogram and Magnetic Resonance Imaging (MRI of cerebro-pontine angle showed no detected abnormality, excluding acoustic neuroma. Thus, we present a case of multiple bilateral isolated cutaneous plexiform schwannomas, not associated with NF2. Multiple plexiform schwannomas is a very rare entity, distinct from neurofibromatosis (NF, and being confined to the dermis is even more rarely reported.

  19. Propellant isolation shutoff valve program

    Science.gov (United States)

    Merritt, F. L.

    1973-01-01

    An analysis and design effort directed to advancing the state-of-the-art of space storable isolation valves for control of flow of the propellants liquid fluorine/hydrazine and Flox/monomethylhydrazine is discussed. Emphasis is on achieving zero liquid leakage and capability of withstanding missions up to 10 years in interplanetary space. Included is a study of all-metal poppet sealing theory, an evaluation of candidate seal configurations, a valve actuator trade-off study and design description of a pneumo-thermally actuated soft metal poppet seal valve. The concepts and analysis leading to the soft seal approach are documented. A theoretical evaluation of seal leakage versus seal loading, related finishes and yield strengths of various materials is provided. Application of a confined soft aluminum seal loaded to 2 to 3 times yield strength is recommended. Use of either an electro-mechanical or pneumatic actuator appears to be feasible for the application.

  20. Bacterial isolates degrading aliphatic polycarbonates.

    Science.gov (United States)

    Suyama, T; Hosoya, H; Tokiwa, Y

    1998-04-15

    Bacteria that degrade an aliphatic polycarbonate, poly(hexamethylene carbonate), were isolated from river water in Ibaraki. Prefecture, Japan, after enrichment in liquid medium containing poly(hexamethylene carbonate) suspensions as carbon source, and dilution to single cells. Four of the strains, 35L, WFF52, 61A and 61B2, degraded poly(hexamethylene carbonate) on agar plate containing suspended poly(hexamethylene carbonate). Degradation of poly(hexamethylene carbonate) was confirmed by gel permeation chromatography. Besides poly(hexamethylene carbonate), the strains were found to degrade poly(tetramethylene carbonate). The strains were characterized morphologically, physiologically, and by 16S rDNA sequence analysis. Strains 35L and WFF52 were tentatively identified as Pseudomonas sp. and Variovorax sp., respectively, while strains 61A and 61B2 constitute an unidentified branch within the beta subclass of the Proteobacteria.

  1. An isolated nasolacrimal duct osteoma.

    Science.gov (United States)

    Kim, Joo Yeon; Kwon, Jae Hwan

    2013-07-01

    Osteomas of the nose and paranasal sinus are common benign tumors that can extend to surrounding structures and result in orbital or intracranial involvement. Presenting symptoms include facial pain, headache, cerebral symptoms, ocular symptoms, and so on, depending on the location and size of the tumor. They commonly occur within the frontal, ethmoid, maxillary, and sphenoid sinuses; however, there are rare cases of reported osteomas in the nasal cavity, turbinate, or orbit. Our case report describes a patient with nasolacrimal duct osteoma who presented with ipsilateral ocular pain, epiphora, and medial canthal swelling. We performed intranasal dacryocystorhinostomy using a nasal endoscope and removed the lacrimal duct osteoma. This report describes symptoms and management of an isolated nasolacrimal duct stone with a review of the literature.

  2. Viruses isolated from Panamanian sloths.

    Science.gov (United States)

    Seymour, C; Peralta, P H; Montgomery, G G

    1983-11-01

    Seven virus strains were isolated in Vero cells from whole blood samples from 80 wild-caught sloths, Bradypus variegatus and Choloepus hoffmanni, from Central Panamá. Four strains of at least two different serotypes are related to Changuinola virus; two of these were associated with prolonged or recrudescent viremias. One strain is an antigenic subtype of Punta Toro virus, and another, described here as Bradypus-4 virus, is a new, antigenically ungrouped virus. A second new virus from sloths, Utive virus, forms an antigenic complex within the Simbu serogroup with Utinga and Pintupo viruses. Tests on sequential plasma samples from radio-marked free-ranging sloths and from recently captured animals maintained in captivity showed that both species develop neutralizing antibodies following naturally acquired virus infections. Antibodies against the Changuinola and Simbu serogroup viruses are widespread in both sloth species and are especially prevalent in Choloepus, but are virtually absent in all other wild vertebrate species tested.

  3. Spectroscopy of Isolated Prebiotic Nucleobases

    Science.gov (United States)

    Svadlenak, Nathan; Callahan, Michael P.; Ligare, Marshall; Gulian, Lisa; Gengeliczki, Zsolt; Nachtigallova, Dana; Hobza, Pavel; deVries, Mattanjah

    2011-01-01

    We use multiphoton ionization and double resonance spectroscopy to study the excited state dynamics of biologically relevant molecules as well as prebiotic nucleobases, isolated in the gas phase. Molecules that are biologically relevant to life today tend to exhibit short excited state lifetimes compared to similar but non-biologically relevant analogs. The mechanism is internal conversion, which may help protect the biologically active molecules from UV damage. This process is governed by conical intersections that depend very strongly on molecular structure. Therefore we have studied purines and pyrimidines with systematic variations of structure, including substitutions, tautomeric forms, and cluster structures that represent different base pair binding motifs. These structural variations also include possible alternate base pairs that may shed light on prebiotic chemistry. With this in mind we have begun to probe the ultrafast dynamics of molecules that exhibit very short excited states and search for evidence of internal conversions.

  4. Cladal relatedness among Aspergillus oryzae isolates and Aspergillus flavus S and L morphotype isolates.

    Science.gov (United States)

    Chang, Perng-Kuang; Ehrlich, Kenneth C; Hua, Sui-Sheng T

    2006-04-25

    Aspergillus flavus is the main etiological agent for aflatoxin contamination of crops. Its close relative, A. oryzae, does not produce aflatoxins and has been widely used to produce fermented foods. We compared the phylogeny of A. oryzae isolates and L- and S-type sclerotial isolates of A. flavus using single nucleotide polymorphisms in the omtA gene in the aflatoxin biosynthesis gene cluster and deletions in and distal to the norB-cypA intergenic region as phylogenetic signals. Aflatoxin-producing ability and sclerotial size also were weighted in the analysis. Like A. flavus, the A. oryzae isolates form a polyphyletic assemblage. A. oryzae isolates in one clade strikingly resemble an A. flavus subgroup of atoxigenic L-type isolates. All toxigenic S-type isolates closely resemble another subgroup of atoxigenic L-type isolates. Because atoxigenic S-type isolates are extremely rare, we hypothesize that loss of aflatoxin production in S-type isolates may occur concomitantly with a change to L-type sclerotia. All toxigenic L-type isolates, unlike A. oryzae, have a 1.0 kb deletion in the norB-cypA region. Although A. oryzae isolates, like S-type, have a 1.5 kb deletion in the norB-cypA region, none were cladally related to S-type A. flavus isolates. Our results show that A. flavus populations are genetically diverse. A. oryzae isolates may descend from certain atoxigenic L-type A. flavus isolates.

  5. Steady compression characteristics of laminated MRE isolator

    Science.gov (United States)

    Wahab, N. A. A.; Mazlan, S. A.; Ubaidillah; Sharif, A. H. R.; Kamaruddin, S.

    2016-11-01

    This paper focused on an experimental setup on laminated magnetorheological elastomer (MRE) isolator under steady state compression test. An isotropic type natural rubber (NR) based MRE were fabricated and layered with a steel plate to form a multilayer sandwich structure adopted from the conventional laminated rubber bearing design. A set of static compression test was conducted to explore the potential of semi-active laminated MRE isolator in field-dependent stiffness properties. Stress versus strain relationship was assessed under different magnetic fields application. Based on the examination, the stress altered as the application of magnetic fields. Consequently, the effective stiffness of isolator also influenced by the magnetic fields induction. The experimental results show that the proposed laminated MRE isolator can effectively alter the compression stiffness up to the 14.56%. The preliminary results have confirmed the tunability of the semi-active laminated MRE isolator in which it would be beneficial for improving building isolator in general.

  6. Isolation and Identification of Concrete Environment Bacteria

    Science.gov (United States)

    Irwan, J. M.; Anneza, L. H.; Othman, N.; Husnul, T.; Alshalif, A. F.

    2016-07-01

    This paper presents the isolation and molecular method for bacteria identification through PCR and DNA sequencing. Identification of the bacteria species is required in order to fully utilize the bacterium capability for precipitation of calcium carbonate in concrete. This process is to enable the addition of suitable catalyst according to the bacterium enzymatic pathway that is known through the bacteria species used. The objective of this study is to isolate, enriched and identify the bacteria species. The bacteria in this study was isolated from fresh urine and acid mine drainage water, Kota Tinggi, Johor. Enrichment of the isolated bacteria was conducted to ensure the bacteria survivability in concrete. The identification of bacteria species was done through polymerase chain reaction (PCR) and rRDNA sequencing. The isolation and enrichment of the bacteria was done successfully. Whereas, the results for bacteria identification showed that the isolated bacteria strains are Bacillus sp and Enterococus faecalis.

  7. Klebsiella pneumoniae KPC: first isolations in Italy

    OpenAIRE

    Carla Fontana; Marco Favaro; Loredana Sarmati; Silvia Natoli; Anna Altieri; Maria Cristina Bossa; Silvia Minelli; Cartesio Favalli

    2009-01-01

    Klebsiella pneumoniae carbapenemase (KPC) was detected in two isolates of carbapenem-resistant K. pneumoniae in an italian teaching hospital. This is the first report of a KPC-producing isolates in our country. The first strain was isolated from a urine sample collected from a indwelling urinary catheter in a ICU-patient with subdural haematoma, while the second was from the culture of the central venous catheter (CVC) in a patient affected by Crohn’s disease admitted in gastroenterology ward...

  8. Isolated trochlear nerve palsy with midbrain hemorrhage

    Directory of Open Access Journals (Sweden)

    Raghavendra S

    2010-01-01

    Full Text Available Midbrain hemorrhage causing isolated fourth nerve palsy is extremely rare. Idiopathic, traumatic and congenital abnormalities are the most common causes of fourth nerve palsy. We report acute isolated fourth nerve palsy in an 18-year-old lady due to a midbrain hemorrhage probably due to a midbrain cavernoma. The case highlights the need for neuroimaging in selected cases of isolated trochlear nerve palsy.

  9. Variable stiffness and damping magnetorheological isolator

    Institute of Scientific and Technical Information of China (English)

    Yang ZHOU; Xingyu WANG; Xianzhou ZHANG; Weihua LI

    2009-01-01

    This paper presents the development and characterization of a magnetorheological (MR) fluid-based variable stiffness and damping isolator. The prototype of the MR fluid isolator is fabricated, and its dynamic behavior is measured under various applied magnetic fields. The parameters of the model under various magnetic fields are identified, and the dynamic perfor-mance of the isolator is evaluated in simulation. Experi-mental results indicate that both the stiffness and damping capability of the developed MR isolator can be controlled by an external magnetic field.

  10. Fault Isolation for Shipboard Decision Support

    DEFF Research Database (Denmark)

    Lajic, Zoran; Blanke, Mogens; Nielsen, Ulrik Dam

    2010-01-01

    Fault detection and fault isolation for in-service decision support systems for marine surface vehicles will be presented in this paper. The stochastic wave elevation and the associated ship responses are modeled in the frequency domain. The paper takes as an example fault isolation of a containe......Fault detection and fault isolation for in-service decision support systems for marine surface vehicles will be presented in this paper. The stochastic wave elevation and the associated ship responses are modeled in the frequency domain. The paper takes as an example fault isolation...

  11. Isolated Fast High-Voltage Switching Circuit

    Science.gov (United States)

    Rizzi, Anthony

    1992-01-01

    Electrically isolated switching circuit supplies pulses at potentials up to 6.5 kV and currents up to 6.5 A, lasting as long as few microseconds. Turn-on time about 40 ns; turn-off time about 3 microseconds. Electrically isolated from control circuitry by means of fiber-optic signal coupling and isolated power supply. Electrical isolation protects both technician and equipment. This and similar circuits useful in such industrial and scientific applications as high-voltage, high-frequency test equipment; electrostatic-discharge test equipment; plasma-laboratory instrumentation; spark chambers; and electromagnetic-interference test equipment.

  12. Hydrologic considerations in defining isolated wetlands

    Science.gov (United States)

    Winter, T.C.; LaBaugh, J.W.

    2003-01-01

    Wetlands that are not connected by streams to other surface-water bodies are considered to be isolated. Although the definition is based on surface-water connections to other water bodies, isolated wetlands commonly are integral parts of extensive ground-water flow systems, and isolated wetlands can spill over their surface divides into adjacent surface-water bodies during periods of abundant precipitation and high water levels. Thus, characteristics of ground-water flow and atmospheric-water flow affect the isolation of wetlands. In general, the degree that isolated wetlands are connected through the ground-water system to other surface-water bodies depends to a large extent on the rate that ground water moves and the rate that hydrologic stresses can be transmitted through the ground-water system. Water that seeps from an isolated wetland into a gravel aquifer can travel many kilometers through the ground-water system in one year. In contrast, water that seeps from an isolated wetland into a clayey or silty substrate may travel less than one meter in one year. For wetlands that can spill over their surface watersheds during periods of wet climate conditions, their isolation is related to the height to a spill elevation above normal wetland water level and the recurrence interval of various magnitudes of precipitation. The concepts presented in this paper indicate that the entire hydrologic system needs to be considered in establishing a definition of hydrologic isolation.

  13. Biodegradation of norfloxacin by Penicillium frequentans isolated ...

    African Journals Online (AJOL)

    PROMOTING ACCESS TO AFRICAN RESEARCH ... One norfloxacin-degrading fungi was isolated from soil contaminated by norfloxacin and preliminary identified as ... Keywords: Norfloxacin, soil, biodegradation, microbial consortium.

  14. Permanent isolation surface barrier: Functional performance

    Energy Technology Data Exchange (ETDEWEB)

    Wing, N.R.

    1993-10-01

    This document presents the functional performance parameters for permanent isolation surface barriers. Permanent isolation surface barriers have been proposed for use at the Hanford Site (and elsewhere) to isolate and dispose of certain types of waste in place. Much of the waste that would be disposed of using in-place isolation techniques is located in subsurface structures, such as solid waste burial grounds, tanks, vaults, and cribs. Unless protected in some way, the wastes could be transported to the accessible environment via transport pathways, such as water infiltration, biointrusion, wind and water erosion, human interference, and/or gaseous release.

  15. Recent advances in micro-vibration isolation

    Science.gov (United States)

    Liu, Chunchuan; Jing, Xingjian; Daley, Steve; Li, Fengming

    2015-05-01

    Micro-vibration caused by disturbance sources onboard spacecraft can severely degrade the working environment of sensitive payloads. Some notable vibration control methods have been developed particularly for the suppression or isolation of micro-vibration over recent decades. Usually, passive isolation techniques are deployed in aerospace engineering. Active isolators, however, are often proposed to deal with the low frequency vibration that is common in spacecraft. Active/passive hybrid isolation has also been effectively used in some spacecraft structures for a number of years. In semi-active isolation systems, the inherent structural performance can be adjusted to deal with variation in the aerospace environment. This latter approach is potentially one of the most practical isolation techniques for micro-vibration isolation tasks. Some emerging advanced vibration isolation methods that exploit the benefits of nonlinearity have also been reported in the literature. This represents an interesting and highly promising approach for solving some challenging problems in the area. This paper serves as a state-of-the-art review of the vibration isolation theory and/or methods which were developed, mainly over the last decade, specifically for or potentially could be used for, micro-vibration control.

  16. Isolated systems with wind power. Main report

    DEFF Research Database (Denmark)

    Lundsager, P.; Bindner, Henrik W.; Clausen, Niels-Erik;

    2001-01-01

    approach for assessing the technical and economical feasibility of isolated power supply systems with wind energy. As a part of the project the following tasks were carried out: Review of literature, fieldmeasurements in Egypt, development of an inventory of small isolated systems, overview of end......The overall objective of this research project is to study the development of methods and guidelines rather than "universal solutions" for the use of wind energy in isolated communities. The main specific objective of the project is to develop and present amore unified and generally applicable...... for Isolated Systems with Wind Power, applicable for international organisations such as donoragencies and development banks....

  17. Blackbody Radiation from Isolated Neptunes

    Science.gov (United States)

    Ginzburg, Sivan; Sari, Re'em; Loeb, Abraham

    2016-05-01

    Recent analyses of the orbits of some Kuiper belt objects hypothesize the presence of an undiscovered Neptune-size planet at a very large separation from the Sun. The energy budget of Neptunes on such distant orbits is dominated by the internal heat released by their cooling rather than solar irradiation (making them effectively “isolated”). The blackbody radiation that these planets emit as they cool may provide the means for their detection. Here, we use an analytical toy model to study the cooling and radiation of isolated Neptunes. This model can translate a detection (or a null detection) to a constraint on the size and composition of the hypothesized “Planet Nine.” Specifically, the thick gas atmosphere of Neptune-like planets serves as an insulating blanket that slows down their cooling. Therefore, a measurement of the blackbody temperature, {T}{{eff}}˜ 50 {{K}}, at which a Neptune emits, can be used to estimate the mass of its atmosphere, {M}{{atm}}. Explicitly, we find the relation {T}{{eff}}\\propto {M}{{atm}}1/12. Despite this weak relation, a measurement of the flux at the Wien tail can constrain the atmospheric mass, at least to within a factor of a few, and provide useful limits to possible formation scenarios of these planets. Finally, we constrain the size and composition of Planet Nine by combining our model with the null results of recent all-sky surveys.

  18. Isolated and Combined Remethylation Disorders

    Directory of Open Access Journals (Sweden)

    Eva Richard PhD

    2017-01-01

    Full Text Available Genetic defects affecting the remethylation pathway cause hyperhomocysteinemia. Isolated remethylation defects are caused by mutations of the 5, 10-methylenetetrahydrofolate reductase (MTHFR, methionine synthase reductase (MTRR, methionine synthase (MTR, and MMADHC genes, and combined remethylation defects are the result of mutations in genes involved in the synthesis of either methylcobalamin or adenosylcobalamin, that is, the active cofactors of MTRR and methylmalonyl-CoA mutase. Diagnosis is based on the biochemical analysis of amino acids, homocysteine, propionylcarnitine, methylmalonic acid, S-adenosylmethionine, and 5-methylentetrahydrofolate in physiological fluids. Gene-by-gene Sanger sequencing has long been the gold standard genetic analysis for confirming the disorder and identifying the gene involved, but massive parallel sequencing is now being used to examine all those potentially involved in one go. Early treatment to rescue metabolic homeostasis is based on the following of an appropriate diet, betaine administration, and, in some cases, oral or intramuscular administration of vitamin B12 or folate. Elevated ROS levels, apoptosis, endoplasmic reticulum (ER stress, the activation of autophagy, and alterations in Ca2+ homeostasis may all contribute toward the pathogenesis of the disease. Pharmacological agents to restore the function of the ER and mitochondria and/or to reduce oxidative stress-induced apoptosis might provide novel ways of treating patients with remethylation disorders.

  19. Single Cell Isolation and Analysis

    Directory of Open Access Journals (Sweden)

    Ping Hu

    2016-10-01

    Full Text Available Increasing evidence shows that the heterogeneity of individual cells within a genetically identical population can be critical to their peculiar function and fate. Conventional cell based assays mainly analysis the average responses from a population cells, while the difference within individual cells may often be masked. The cell size, RNA transcripts and protein expression level are quite different within individual cells and these variations are key point to answer the problems in cancer, neurobiology, stem cell biology, immunology and developmental biology. To better understand the cell-to-cell variations, the single cell analysis can provide much more detailed information which may be helpful for therapeutic decisions in an increasingly personalized medicine. In this review, we will focus on the recent development in single cell analysis, including methods used in single cell isolation, analysis and some application examples. The review provides the historical background to single cell analysis, discusses limitations, and current and future possibilities in this exciting field of research.

  20. Isolated and Combined Remethylation Disorders

    Directory of Open Access Journals (Sweden)

    Eva Richard PhD

    2017-01-01

    Full Text Available Genetic defects affecting the remethylation pathway cause hyperhomocysteinemia. Isolated remethylation defects are caused by mutations of the 5, 10-methylenetetrahydrofolate reductase (MTHFR , methionine synthase reductase (MTRR , methionine synthase (MTR , and MMADHC genes, and combined remethylation defects are the result of mutations in genes involved in the synthesis of either methylcobalamin or adenosylcobalamin, that is, the active cofactors of MTRR and methylmalonyl-CoA mutase. Diagnosis is based on the biochemical analysis of amino acids, homocysteine, propionylcarnitine, methylmalonic acid, S-adenosylmethionine, and 5-methylentetrahydrofolate in physiological fluids. Gene-by-gene Sanger sequencing has long been the gold standard genetic analysis for confirming the disorder and identifying the gene involved, but massive parallel sequencing is now being used to examine all those potentially involved in one go. Early treatment to rescue metabolic homeostasis is based on the following of an appropriate diet, betaine administration, and, in some cases, oral or intramuscular administration of vitamin B 12 or folate. Elevated ROS levels, apoptosis, endoplasmic reticulum (ER stress, the activation of autophagy, and alterations in Ca 2+ homeostasis may all contribute toward the pathogenesis of the disease. Pharmacological agents to restore the function of the ER and mitochondria and/or to reduce oxidative stress-induced apoptosis might provide novel ways of treating patients with remethylation disorders.

  1. Isolated Pancreatic Uncinate Duct IPMN.

    Science.gov (United States)

    Maker, Ajay V; Maker, Vijay K

    2017-04-01

    The ventral pancreas originally forms as an evagination of the common bile duct at 32 days gestation and its duct, the uncinate duct, eventually rotates with the ventral anlage to join the dorsal pancreas and fuse with the main pancreatic duct. Thus, though often considered a "branch" duct of the pancreas, embryologically, the uncinate duct is the "main" pancreatic duct of the ventral pancreas. This concept is not fully addressed in the current definitions of intraductal papillary mucinous neoplasms of the pancreas (IPMN) where international consensus guidelines consider the main-duct IPMN as high risk for malignancy and most small branch-duct IPMN as low risk for malignancy. Thus, it is important to recognize that isolated uncinate-duct IPMN can occur and, based on its embryologic origin and increased association with high-grade dysplasia and invasive cancer, may be managed conceptually as a main duct type of disease rather than a branch duct until better biomarkers of malignancy are discovered. The images provide an example of this unique disease process.

  2. Fault Detection and Isolation for Spacecraft

    DEFF Research Database (Denmark)

    Jensen, Hans-Christian Becker; Wisniewski, Rafal

    2002-01-01

    This article realizes nonlinear Fault Detection and Isolation for actuators, given there is no measurement of the states in the actuators. The Fault Detection and Isolation of the actuators is instead based on angular velocity measurement of the spacecraft and knowledge about the dynamics...

  3. Isolation and characterization of extreme halophilic archaea

    Energy Technology Data Exchange (ETDEWEB)

    Franze, Madlen; Cherkouk, Andrea [Helmholtz-Zentrum Dresden-Rossendorf e.V., Dresden (Germany). HZDR Young Investigator Group

    2017-06-01

    Extreme halophilic archaea from the family Halobactereacea represent a dominant part of the microbial community present in saline soils as well as rock salts. By using a culture-dependent approach different Haloarchaea could be isolated and were phylogenetic analysed. Interestingly, isolates closely related to different Halobacterium spp. were found in both environments.

  4. Scalable Detection and Isolation of Phishing

    NARCIS (Netherlands)

    Moreira Moura, G.C.; Pras, A.

    2009-01-01

    This paper presents a proposal for scalable detection and isolation of phishing. The main ideas are to move the protection from end users towards the network provider and to employ the novel bad neighborhood concept, in order to detect and isolate both phishing e-mail senders and phishing web server

  5. Decode Fish Lymphocystis Virus Isolated from China

    Institute of Scientific and Technical Information of China (English)

    2004-01-01

    @@ Aresearch group headed by Prof. Zhang Qiya from the CAS Institute of Hydrobiology (IHB) has succeeded in sequencing the complete genome of lymphocystis disease virus isolated from China (LCDV-C), a virus isolated from cultured flounder (Paralichthys olivaceus) with lymphocystis disease in China. Their work has been published in the recent issue of Journal of Virology (2004, 78 (13): 6982- 6994).

  6. Epidemic model with isolation in multilayer networks

    CERN Document Server

    Zuzek, L G Alvarez; Braunstein, L A

    2014-01-01

    The Susceptible-Infected-Recovered (SIR) model has successfully mimicked the propagation of such airborne diseases as influenza A (H1N1). Although the SIR model has recently been studied in a multilayer networks configuration, in almost all the research the dynamic movement of infected individuals, e.g., how they are often kept in isolation, is disregarded. We study the SIR model in two multilayer networks and use an isolation parameter, indicating time period, to measure the effect of isolating infected individuals from both layers. This isolation reduces the transmission of the disease because the time in which infection can spread is reduced. In this scenario we find that the epidemic threshold increases with the isolation time and the isolation parameter and the impact of the propagation is reduced. We also find that when isolation is total there is a threshold for the isolation parameter above which the disease never becomes an epidemic. We also find that regular epidemic models always overestimate the e...

  7. Professional Isolation and Stress in Teachers.

    Science.gov (United States)

    Dussault, Marc; And Others

    The aim of this study was to analyze the relationship between professional isolation and occupational stress in teachers. A systematic random sample of 1158 French Canadian teachers were administered French Canadian versions of the "UCLA Loneliness Scale and Teacher Stress Inventory." Professional isolation was measured by the subjects'…

  8. Gene Diversity of Trichomonas Vaginalis Isolates

    Directory of Open Access Journals (Sweden)

    Z Valadkhani

    2011-09-01

    Full Text Available Background: Trichomonas vaginalis is protozoan parasite responsible for trichomoniasis and is more common in high-risk behavior group such as prostitute individuals. Interest in trichomoni­asis is due to increase one's susceptibility to viruses such as herpes, human papillomavirus and HIV. The aim of this study was to find genotypic differences between the isolates.Methods: Forty isolates from prisoners' women in Tehran province were used in this study. The random amplified polymorphic DNA (RAPD technique was used to determine genetic differ­ences among isolates and was correlated with patient's records. By each primer the banding pat­tern size of each isolates was scored (bp, genetic differences were studied, and the genealogical tree was constructed by using NTSYS software program and UPGMA method.Results: The least number of bands were seen by using primer OPD8 and the most by using OPD3. Results showed no significant difference in isolates from different geographical areas in Iran. By using primer OPD1 specific amplified fragment with length 1300 base pair were found in only 8 isolates. All these isolates were belonged to addicted women; however, six belonged to asymptomatic patients and two to symptomatic ones.Conclusion: There was not much genetic diversity in T vaginalis isolates from three different geo­graphical areas.

  9. Understanding the differences in gelling properties between lupin protein isolate and soy protein isolate

    NARCIS (Netherlands)

    Berghout, J.A.M.; Boom, R.M.; Goot, van der A.J.

    2015-01-01

    The gelling properties of lupin protein isolate (LPI) were compared with those of soy protein isolate (SPI). It was found that LPI behaves fundamentally different than SPI, evidenced by the formation of weaker and deformable gels. Further investigation shows that both protein isolates can be conside

  10. Sexually dimorphic neuronal responses to social isolation

    Science.gov (United States)

    Senst, Laura; Baimoukhametova, Dinara; Sterley, Toni-Lee; Bains, Jaideep Singh

    2016-01-01

    Many species use social networks to buffer the effects of stress. The mere absence of a social network, however, may also be stressful. We examined neuroendocrine, PVN CRH neurons and report that social isolation alters the intrinsic properties of these cells in sexually dimorphic fashion. Specifically, isolating preadolescent female mice from littermates for neurons. These changes were not evident in age-matched males. By contrast, subjecting either males (isolated or grouped) or group housed females to acute physical stress (swim), increased FSL. The increase in FSL following either social isolation or acute physical stress was blocked by the glucocorticoid synthesis inhibitor, metyrapone and mimicked by exogenous corticosterone. The increase in FSL results in a decrease in the excitability of CRH neurons. Our observations demonstrate that social isolation, but not acute physical stress has sex-specific effects on PVN CRH neurons. DOI: http://dx.doi.org/10.7554/eLife.18726.001 PMID:27725087

  11. Isolation of Pasteurella multocida from broiler chickens

    Directory of Open Access Journals (Sweden)

    Sri Poernomo

    1996-06-01

    Full Text Available Pasteurella multocida, the etiological agent of fowl cholera, was isolated from five, 32 days oldbroilerchickens in the late of 1992. The chickens were from a farm located in Bogor area, raised in cages and each flock consisted of 1,550 broilers . Therewere 230 birds, aging from 28-31 days old, died with clinical signs of lameness and difficulty in breathing. Serological test of the isolate revealed serotype Aof Carter classification . To prove its virulences, the isolate was then inoculated into 3 mice subcutaneously. The mice died less then 24 hours postinoculation and P. multocida can be reisolated . The sensitivity test to antibiotics and sulfa preparations showed that the isolate was sensitive to ampicillin, doxycyclin, erythromycin, gentamycin, sulfamethoxazol-trimethoprim and baytril, but resistance to tetracyclin, kanamycin and oxytetracyclin. This is the first report of P. multocida isolation in broiler chickens in Indonesia, and it is intended to add information on bacterial diseases in poultry in Indonesia.

  12. Listeria fleischmannii sp. nov., isolated from cheese.

    Science.gov (United States)

    Bertsch, David; Rau, Jörg; Eugster, Marcel R; Haug, Martina C; Lawson, Paul A; Lacroix, Christophe; Meile, Leo

    2013-02-01

    A study was performed on three isolates (LU2006-1(T), LU2006-2 and LU2006-3), which were sampled independently from cheese in western Switzerland in 2006, as well as a fourth isolate (A11-3426), which was detected in 2011, using a polyphasic approach. The isolates could all be assigned to the genus Listeria but not to any known species. Phenotypic and chemotaxonomic data were compatible with the genus Listeria and phylogenetic analysis based on 16S rRNA gene sequences confirmed that the closest relationships were with members of this genus. However, DNA-DNA hybridization demonstrated that the isolates did not belong to any currently described species. Cell-wall-binding domains of Listeria monocytogenes bacteriophage endolysins were able to attach to the isolates, confirming their tight relatedness to the genus Listeria. Although PCR targeting the central portion of the flagellin gene flaA was positive, motility was not observed. The four isolates could not be discriminated by Fourier transform infrared spectroscopy or pulsed-field gel electrophoresis. This suggests that they represent a single species, which seems to be adapted to the environment in a cheese-ripening cellar as it was re-isolated from the same type of Swiss cheese after more than 5 years. Conjugation experiments demonstrated that the isolates harbour a transferable resistance to clindamycin. The isolates did not exhibit haemolysis or show any indication of human pathogenicity or virulence. The four isolates are affiliated with the genus Listeria but can be differentiated from all described members of the genus Listeria and therefore they merit being classified as representatives of a novel species, for which we propose the name Listeria fleischmannii sp. nov.; the type strain is LU2006-1(T) ( = DSM 24998(T)  = LMG 26584(T)).

  13. Staphylococcus epidermidis isolated in 1965 are more susceptible to triclosan than current isolates

    DEFF Research Database (Denmark)

    Skovgaard, Sissel; Nielsen, Lene Nørby; Larsen, Marianne Halberg;

    2013-01-01

    pathogen Staphylococcus epidermidis with isolates collected in the 1960s prior to introduction of triclosan to the market. Of 64 current S. epidermidis isolates 12.5% were found to have tolerance towards triclosan defined as MIC≥0.25 mg/l compared to none of 34 isolates obtained in the 1960s. When passaged...... in the laboratory in the presence of triclosan, old and current susceptible isolates could be adapted to the same triclosan MIC level as found in current tolerant isolates. DNA sequence analysis revealed that laboratory-adapted strains carried mutations in fabI encoding the enoyl-acyl carrier protein reductase...

  14. Isolation of nuc mutant isolates of Staphylococcus aureus from bovine clinical mastitis.

    Science.gov (United States)

    Zastempowska, E; Orczykowska-Kotyna, M; Lassa, H

    2014-06-01

    Isolates of Staphylococcus aureus with a mutation in the nuclease (nuc) gene were recovered from cases of bovine mastitis in Poland. Three S. aureus isolates from cows in one herd had a 42 base pair duplication in the nuc gene. These isolates belonged to sequence type 97 (ST97) and clonal complex 97 (CC97). They had a different spa type and multiple-locus variable-number tandem-repeat fingerprinting (MLVF) subtype than a S. aureus isolate without the nuc mutation from the same herd. Isolation of nuc mutant S. aureus strains from cases of bovine mastitis may confound diagnostic PCRs based on detection of the nuc gene.

  15. Molecular characterization of mycobacteria isolated from seals.

    Science.gov (United States)

    Zumárraga, M J; Bernardelli, A; Bastida, R; Quse, V; Loureiro, J; Cataldi, A; Bigi, F; Alito, A; Castro Ramos, M; Samper, S; Otal, I; Martin, C; Romano, M I

    1999-09-01

    Tuberculosis (TB) was diagnosed in 10 seals from three species (Arctocephalus australis, Arctocephalus tropicalis and Otaria flavescens) found in South America. The mycobacteria isolated from these cases belonged to the Mycobacterium tuberculosis complex, as determined by RFLP using an IS6110 probe, spoligotyping, analysis of the 16S rRNA gene sequence and by PCR-restriction analysis of hsp65. Polymorphisms in gyrA, katG, oxyR and pncA were investigated in some of the isolates, as well as the presence of the MPB70 antigen. The insertion sequence IS6110 was present in three to seven copies in the genome of the mycobacteria isolated from seals. Using the IS6110 probe, six patterns (designated A, B, C, D, E and F) were identified from 10 different isolates. Patterns A and B were found for the mycobacteria isolated from two and four seals, respectively, indicating an epidemiological relationship between isolates grouped according to their IS6110 RFLP. The mycobacteria isolated from seals shared the majority of their IS6110 DNA-containing restriction fragments, and nine isolates had an identical spoligotype; only one isolate showed a minor difference in its spoligotype. In addition, none of these spoligotypes were found in other M. tuberculosis complex strains. These results suggest that the isolates from seals constitute a unique group of closely related strains. The mycobacteria isolated from seals showed polymorphisms at gyrA codon 95 and katG codon 463, as do group 1 M. tuberculosis, and M. bovis. Group 1 mycobacteria are associated with cluster cases. The spoligotypes found in the mycobacteria isolated from seals lack spacers 39-43, as does M. bovis, but the MPB70 antigen, which is highly expressed in M. bovis and minimally expressed in M. tuberculosis, was not detected in these mycobacteria. The mycobacteria isolated from seals also showed oxyR and pncA polymorphisms specific to M. tuberculosis. In conclusion, the mycobacteria that cause TB in seals in the South

  16. Microbiology of a wetland ecosystem constructed to remediate mine drainage from a heavy metal mine.

    Science.gov (United States)

    Hallberg, Kevin B; Johnson, D Barrie

    2005-02-01

    A pilot passive treatment plant (PPTP) was constructed to evaluate the potential of a composite wetland system to remediate acidic, metal-rich water draining the former Wheal Jane tin, in Cornwall, England. The treatment plant consists of three separate and controllable composite systems, each of which comprises a series of aerobic wetlands for iron oxidation and precipitation, a compost bioreactor for removing chalcophilic metals and to generate alkalinity, and rock filter ponds for removing soluble manganese and organic carbon. To understand the roles of microorganisms in remediating acid mine drainage (AMD) in constructed wetland ecosystems, populations of different groups of cultivatable acidophilic microbes in the various components of the Wheal Jane PPTP were enumerated over a 30-month period. Initially, moderately acidophilic iron-oxidising bacteria (related to Halothiobacillus neapolitanus) were found to be the major cultivatable microorganisms present in the untreated AMD, though later heterotrophic acidophiles emerged as the dominant group, on a numerical basis. Culturable microbes in the surface waters and sediments of the aerobic wetlands were similarly dominated by heterotrophic acidophiles, though both moderately and extremely acidophilic iron-oxidising bacteria were also present in significant numbers. The dominant microbial isolate in waters draining the anaerobic compost bioreactors was an iron- and sulfur-oxidising moderate acidophile that was closely related to Thiomonas intermedia. The acidophiles enumerated at the Wheal Jane PPTP accounted for 1% to 25% of the total microbial population. Phylogenetic analysis of 14 isolates from various components of the Wheal Jane PPTP showed that, whilst many of these bacteria were commonly encountered acidophiles, some of these had not been previously encountered in AMD and AMD-impacted environments.

  17. Genetic and phylogenetic analysis of feline calicivirus isolates in China.

    Science.gov (United States)

    Sun, Yaxin; Deng, Mingliang; Peng, Zhong; Hu, Ruiming; Chen, Huanchun; Wu, Bin

    2017-02-01

    The aim of this study was to determine the genetic diversity of Chinese feline calicivirus (FCV) isolates and their phylogenetic relationship with isolates from elsewhere in the world. Phylogenetic analysis was performed based on the partial open reading frame (ORF) 2 sequences (regions B-F) of 21 Chinese FCV isolates and 30 global isolates. The Chinese isolates included 13 isolates from Wuhan, which were isolated in this study, and eight previously published isolates. Sixteen Chinese isolates and two Japanese isolates formed a distinct phylogenetic cluster. Phylogenetic analysis based on the sequences of the complete genome, ORF1, ORF2 and ORF3 of selected isolates supported the above findings. Genogroup analysis revealed that FCV genogroup II is present in China. These findings suggest that Chinese FCV isolates are closely related to Japanese FCV isolates. Copyright © 2016 Elsevier Ltd. All rights reserved.

  18. Are Isolated Indigenous Populations Headed toward Extinction?

    Directory of Open Access Journals (Sweden)

    Robert S Walker

    Full Text Available At least 50 indigenous groups spread across lowland South America remain isolated and have only intermittent and mostly hostile interactions with the outside world. Except in emergency situations, the current policy of governments in Brazil, Colombia, and Peru towards isolated tribes is a "leave them alone" strategy, in which isolated groups are left uncontacted. However, these no-contact policies are based on the assumption that isolated populations are healthy and capable of persisting in the face of mounting external threats, and that they can maintain population viability in the long-term. Here, we test this assumption by tracking the sizes and movements of cleared horticultural areas made by 8 isolated groups over the last 10-14 years. We used deforestation data derived from remote sensing Landsat satellite sensors to identify clearings, and those were then validated and assessed with high-resolution imagery. We found only a single example of a relatively large and growing population (c. 50 cleared ha and 400 people, whereas all of the other 7 groups exhibited much smaller villages and gardens with no sizable growth through time. These results indicated that the smaller groups are critically endangered, and it prompts an urgent re-thinking of policies toward isolated populations, including plans for well-organized contacts that may help save lives and rescue isolated indigenous populations from imminent extinction.

  19. Behavioural reproductive isolation and speciation in Drosophila

    Indian Academy of Sciences (India)

    Punita Nanda; Bashisth Narayan Singh

    2012-06-01

    The origin of premating reproductive isolation continues to help elucidate the process of speciation and is the central event in the evolution of biological species. Therefore, during the process of species formation the diverging populations must acquire some means of reproductive isolation so that the genes from one gene pool are prevented from dispersing freely into a foreign gene pool. In the genus Drosophila, the phenomenon of behavioural reproductive isolation, which is an important type of premating (prezygotic) reproductive isolating mechanisms, has been extensively studied and interesting data have been documented. In many cases incomplete sexual isolation has been observed and the pattern and degree of isolation within and between the species have often been used to elucidate the phylogenetic relationships. The present review documents an overview of speciation mediated through behavioural incompatibility in different species groups of Drosophila with particular reference to the models proposed on the basis of one-sided ethological isolation to predict the direction of evolution. This study is crucial for understanding the mechanism of speciation through behavioural incompatibility and also for an understanding of speciation genetics in future prospects.

  20. Seismic isolation of nuclear power plants

    Energy Technology Data Exchange (ETDEWEB)

    Whittaker, Andrew S.; Kuma, Manish [Dept. of Civil, Structural and Environmental Engineering, State University of New York, Buffalo (United States)

    2014-10-15

    Seismic isolation is a viable strategy for protecting safety-related nuclear structures from the effects of moderate to severe earthquake shaking. Although seismic isolation has been deployed in nuclear structures in France and South Africa, it has not seen widespread use because of limited new build nuclear construction in the past 30 years and a lack of guidelines, codes and standards for the analysis, design and construction of isolation systems specific to nuclear structures. The funding by the United States Nuclear Regulatory Commission of a research project to the Lawrence Berkeley National Laboratory and MCEER/University at Buffalo facilitated the writing of a soon-to-be-published NUREG on seismic isolation. Funding of MCEER by the National Science Foundation led to research products that provide the technical basis for a new section in ASCE Standard 4 on the seismic isolation of safety-related nuclear facilities. The performance expectations identified in the NUREG and ASCE 4 for seismic isolation systems, and superstructures and substructures are described in the paper. Robust numerical models capable of capturing isolator behaviors under extreme loadings, which have been verified and validated following ASME protocols, and implemented in the open source code OpenSees, are introduced.

  1. Insulin Secretory Defect and Insulin Resistance in Isolated Impaired Fasting Glucose and Isolated Impaired Glucose Tolerance

    Directory of Open Access Journals (Sweden)

    Sae Aoyama-Sasabe

    2016-01-01

    Full Text Available Objective. To investigate the characteristics of isolated impaired glucose tolerance (IGT and isolated impaired fasting glucose (IFG, we analyzed the factors responsible for elevation of 2-hour postchallenge plasma glucose (2 h PG and fasting plasma glucose (FPG levels. Methods. We investigated the relationship between 2 h PG and FPG levels who underwent 75 g OGTT in 5620 Japanese subjects at initial examination for medical check-up. We compared clinical characteristics between isolated IGT and isolated IFG and analyzed the relationships of 2 h PG and FPG with clinical characteristics, the indices of insulin secretory capacity, and insulin sensitivity. Results. In a comparison between isolated IGT and isolated IFG, insulinogenic index was lower in isolated IGT than that of isolated IFG (0.43 ± 0.34 versus 0.50 ± 0.47, resp.; p<0.01. ISI composite was lower in isolated IFG than that of isolated IGT (6.87 ± 3.38 versus 7.98 ± 4.03, resp.; p<0.0001. In isolated IGT group, insulinogenic index showed a significant correlation with 2 h PG (r=-0.245, p<0.0001 and had the strongest correlation with 2 h PG (β=-0.290. In isolated IFG group, ISI composite showed a significant correlation with FPG (r=-0.162, p<0.0001 and had the strongest correlation with FPG (β=-0.214. Conclusions. We have elucidated that decreased early-phase insulin secretion is the most important factor responsible for elevation of 2 h PG levels in isolated IGT subjects, and decreased insulin sensitivity is the most important factor responsible for elevation of FPG levels in isolated IFG subjects.

  2. Isolation and Identifaction of Acidithiobacillus thiooxidans with Strong Phosphorous Ore Bioleaching Ability%一株高浸磷嗜酸氧化硫硫杆菌的分离与鉴定

    Institute of Scientific and Technical Information of China (English)

    吕早生; 关海燕; 李凌凌; 贾伟伟

    2011-01-01

    从矿山土样中分离到一株嗜酸的浸磷矿细菌HY-01,观察其形态并研究其培养特征.结果显示,该菌株为革兰氏阴性、短杆状运动细菌,菌体长1~2μm,宽0.5~0.7 μm,能在pH 1.5~8.0的范围内生长,最适生长温度为37℃,最适pH为1.5~3.0.该菌株可以KNO3、NH4NO3、(NH4)2CO3、NH4H2PO4、(NH4)2SO4以及蛋白胨为氮源,以元素硫和黄铁矿中的硫为能源,不能利用有机物质以及硫代硫酸钠、硫酸亚铁等无机物为能源生长.形态、生理生化特性研究以及16SrDNA序列分析表明,该菌株为嗜酸氧化硫硫杆菌.浸矿实验结果表明其具有很高的磷矿浸出能力.图6表1参18%Acidophilic sulfur-oxidizing bacterium HY-01 was isolated from mine soil. Strain HY-01 was motile, gram-negative, rod-shaped and 0.5 to 0.7 μm x 1 to 2 um in size. The strain grew optimally with initial pH 2.0-2.5 and temperature 37 °C. It could grow with potassium nitrate, ammonium nitrate, ammonirm carbonate, ammonium dihydrogen phosphate, ammonium sulfate and tryptone as nitrogen sources. It could use elemental sulfur and pyrite as energy sources, but not organic minerals and inorganic minerals, including sodium thiosulfate and ferrous sulfate. The morphological, biochemical and physiological characteristics of the strain and the analysis based on 16S rDNA gene sequence indicated that strain HY-01 was most closely related to Acidithiobacillus thiooxidans. The sulfur-oxidizing bacteria could positively increase the dissolution rate and percentage of phosphorus recovery from phosphate ore. Fig 6, Tab 1, Ref 18

  3. 三株氧化硫硫杆菌的分离与鉴定%Isolation and Characterization of three strains of Acidithiobacillus thiooxidans and its bioleaching of chalcopyrite

    Institute of Scientific and Technical Information of China (English)

    符波; 周洪波; 张倩; 邱冠周

    2008-01-01

    Three strains of mesophilic and acidophilic sulfur-oxidizing bacteria were isolated from coal heap drainage. The bacteria are motile, Gram-negative, rod-shaped, and measures 0.4 to 0.7 by 1 to 2 μm. They grow optimally at 30 ℃ and initial pH 2.0~2.5. The strains grow autotrophically by using elemental sulfur, sodium thiosulfate and potassium tetrathionate as energy sources. The strains can not use organic matter and inorganic minerals including ferrous sulfate, pyrite and chalcopyrite as energy sources. The morphological, biochemical and physiological characterization and analysis based on 16S rRNA gene sequence indicated that the strains are most closely related to Acidithiobacillus thiooxidans (>99% similarity in gene sequence). The presence of sulfur-oxidizing bacteria positively increased the dissolution rate and the percentage recovery of copper from chalcopyrite by producing acid.%从煤堆废水中分离得到3株嗜温嗜酸硫氧化细菌.这3株菌株为革兰氏阴性、菌体大小0.4~0.7 μm×1~2 μm、短杆状运动细菌,其最适生长温度为 30 ℃和最适生长pH 2.0~2.5.它们能够利用元素硫,硫代硫酸钠和连四硫酸钾为能源进行自养生长,不能利用有机物质以及硫酸亚铁、黄铁矿和黄铜矿等无机物质作为能源生长.细菌的形态、生理生化特性研究以及基于16S rRNA序列同源性构建的系统发育树结果表明,这3株细菌初步鉴定为氧化硫硫杆菌.氧化硫硫杆菌能够通过产酸有效促进黄铜矿的浸出速率和浸出率.

  4. Genetic differences between avian and human isolates of Candida dubliniensis.

    LENUS (Irish Health Repository)

    McManus, Brenda A

    2009-09-01

    When Candida dubliniensis isolates obtained from seabird excrement and from humans in Ireland were compared by using multilocus sequence typing, 13 of 14 avian isolates were genetically distinct from human isolates. The remaining avian isolate was indistinguishable from a human isolate, suggesting that transmission may occur between humans and birds.

  5. Isolation of Klebsiella terrigena from clinical specimens.

    Science.gov (United States)

    Podschun, R; Ullmann, U

    1992-04-01

    In a three-year survey conducted from 1988 to 1990 Klebsiella isolates from human clinical specimens were subjected to additional tests to identify any Klebsiella terrigena strains. Ten strains of Klebsiella terrigena (0.4%) were found among 2355 indole-negative Klebsiella isolates. Most of the isolates were recovered from the respiratory tract. In the API20EC system almost exclusively biotypes no. 1777771 and 1777671 were observed. Serotyping revealed capsule types K2, K5 and K18 in two strains each. In antibiotic susceptibility tests the strains were shown to be comparable in sensitivity to Klebsiella pneumoniae.

  6. Plasmid profiles of Moraxella bovis isolates.

    Science.gov (United States)

    McDonald, T J; Pugh, G W

    1986-04-01

    Two-hundred isolates of Moraxella bovis were selected at random and examined for the presence of plasmid DNA by a rapid alkaline-detergent lysis method. All isolates contained from 1 to 6 plasmids, with varying agarose-gel electrophoretic migration patterns. Most (80%) isolates carried 2 to 4 plasmids, which ranged in molecular weight from 2.6 to 80 megadaltons. Seemingly, plasmid profiles can be used as a simple, reliable epizootiologic tool to establish a strain identification scheme for M bovis.

  7. Isolation of Chloroplasts from Plant Protoplasts.

    Science.gov (United States)

    Lung, Shiu-Cheung; Smith, Matthew D; Chuong, Simon D X

    2015-10-01

    Chloroplasts can be isolated from higher plants directly following homogenization; however, the resulting yield, purity, and intactness are often low, necessitating a large amount of starting material. This protocol is optimized to produce a high yield of pure chloroplasts from isolated Arabidopsis protoplasts. The two-part method is a simple, scaled-down, and low-cost procedure that readily provides healthy mesophyll protoplasts, which are then ruptured to release intact chloroplasts. Chloroplasts isolated using this method are competent for use in biochemical, cellular, and molecular analyses.

  8. Isolated Galaxies in Observations and Simulations

    Science.gov (United States)

    Pilipenko, S.; Doroshkevich, A.

    2010-10-01

    A comparison of observed and simulated isolated galaxies allows us to check their evolution in a wide range of redshifts and to reveal and evaluate a possible impact of their environment. Using the Minimal Spanning Tree (MST) technique we select samples of isolated galaxies, in the SDSS DR7 and in the moderate resolution dark matter simulation. We find that the separation to the nearest neighbour should be 4-5 h-1 Mpc to produce a sample of isolated objects with substantially peculiar environment.

  9. ISOLATION OF CLOSTRIDIUM TETANI FROM SOIL.

    Science.gov (United States)

    SANADA, I; NISHIDA, S

    1965-03-01

    Sanada, Ichiro (Kanazawa University, Kanazawa, Japan), and Shoki Nishida. Isolation of Clostridium tetani from soil. J. Bacteriol. 89:626-629. 1965.-The higher the temperatures applied to soil specimens, the weaker the toxigenicity of Clostridium tetani strains isolated from them. The glucose- and maltose-fermenting ability of these isolates was inversely proportional to their toxigenicity. The biological properties of atoxic strains were indistinguishable from those of C. tetanomorphum. Since a considerable number of toxic strains fermented glucose and maltose, these criteria are of doubtful value for differentiating C. tetani from C. tetanomorphum.

  10. Isolation of Leptospira noguchii from sheep.

    Science.gov (United States)

    Silva, Everton F; Brod, Claudiomar S; Cerqueira, Gustavo M; Bourscheidt, Débora; Seyffert, Núbia; Queiroz, Adriano; Santos, Cleiton S; Ko, Albert I; Dellagostin, Odir A

    2007-03-31

    The main goal of this study was to obtain new isolates of Leptospira spp. from sheep. A total of 10 kidney samples and 44 blood samples were collected from sheep slaughtered in Pelotas, Southern Brazil. One isolate was obtained which was identified by 16S rRNA gene sequencing and serogrouping to be Leptospira noguchii serogroup Autumnalis. Microscopic agglutination test (MAT) evaluation revealed that 4.5% of the sheep sera reacted against the Autumnalis serogroup. This is the first report of isolation of L. noguchii from sheep. Together these findings indicate that L. noguchii infections may be a potentially important veterinary problem in this domestic animal species.

  11. Protein distribution in lupin protein isolates from Lupinus angustifolius L. prepared by various isolation techniques.

    Science.gov (United States)

    Muranyi, Isabel S; Volke, Daniela; Hoffmann, Ralf; Eisner, Peter; Herfellner, Thomas; Brunnbauer, Markus; Koehler, Peter; Schweiggert-Weisz, Ute

    2016-09-15

    Differences in the protein distribution of various protein isolates from Lupinus angustifolius L. Vitabor were identified as affected by the isolation procedure (alkaline and/or salt-induced extraction followed by isoelectric and/or dilutive precipitation). Protein isolates extracted in alkaline solution showed higher protein yields (26.4-31.7%) compared to salt-induced extraction (19.8-30.0%) or combined alkaline and salt-induced extraction (23.3-25.6%). Chemical variations among the protein isolates especially occurred within the albumins. Protein isolates precipitated isoelectrically showed the highest contents, whereas protein isolates precipitated by dilutive showed the lowest contents of conglutin δ. Furthermore, the alkaline subunits of conglutin α and conglutin γ decreased during alkaline extraction compared to salt-induced extraction. A decrease in protein-bound polar and basic amino acids was shown after protein isolation. In contrast, the amounts of nonpolar, aliphatic, aromatic, hydroxylated and sulfur-rich amino acids were higher in the lupin protein isolates compared to the lupin flakes. However, the functional side chains could not be related to the specific molecular arrangements of the protein isolates, as a similar amino acid composition was found among the protein isolates. Copyright © 2016 Elsevier Ltd. All rights reserved.

  12. A case of familial isolated hemihyperplasia

    Directory of Open Access Journals (Sweden)

    Bacino Carlos A

    2004-02-01

    Full Text Available Abstract Background Hemihyperplasia (hemihypertrophy is defined as asymmetric body overgrowth of one or more body parts. Hemihyperplasia can be isolated or be part of well-defined syndromes such as in the case of Beckwith-Wiedemann syndrome (BWS. Isolated hemihyperplasia is usually sporadic, but a number of familial occurrences have been described. Case presentation We describe a Tunisian family in which three maternal cousins and their maternal grandfather present with isolated hemihyperplasia. Conclusions The etiology of isolated hemihyperplasia is unknown although in BWS, genomic imprinting has been shown to play a role in the asymmetric overgrowth. Given the similarity between these two conditions, it is possible that both may share a common pathogenesis. We also discuss the possible genetic mechanisms leading to the production of hemihyperplasia in this family.

  13. Fault Detection and Isolation using Eigenstructure Assignment

    DEFF Research Database (Denmark)

    Jørgensen, R.B.; Patton, R.J.; Chen, J.

    1994-01-01

    The purpose of this article is to investigate the robustness to model uncertainties of observer based fault detection and isolation. The approach is designed with a straight forward dynamic nad the observer....

  14. Thermolysin in human cultured keratinocyte isolation

    Directory of Open Access Journals (Sweden)

    A. Gragnani

    Full Text Available BACKGROUND: When treating extensively burned patients using cultured epidermal sheets, the main problem is the time required for its production. Conventional keratinocyte isolation is usually done using Trypsin. We used a modification of the conventional isolation method in order to improve this process and increase the number of colonies from the isolated epidermal cell population. PURPOSE: To compare the action of trypsin and thermolysin in the keratinocyte isolation using newborn foreskin. METHODS: This method used thermolysin as it selectively digests the dermo-epidermal junction. After dermis separation, the epidermis was digested by trypsin in order to obtain a cell suspension. RESULTS: Compared to the conventional procedure, these experiments demonstrated that in the thermolysin group, the epidermis was easily detached from the dermis, there was no fibroblast contamination and there were a larger number of keratinocyte colonies which had a significant statistical difference. CONCLUSION: The number of colonies in the thermolysin group was significantly greater than in the trypsin group.

  15. Antigenic typing Polish isolates of canine parvovirus

    Energy Technology Data Exchange (ETDEWEB)

    Mizak, B. [National Veterinary Research Institute, Pulawy (Poland); Plucienniczak, A. [Polish Academy ofd Sciences. Microbiology and Virology Center, Lodz (Poland)

    1995-12-31

    Polish strains of canine parvovirus isolated between 1982 and 1993 were examined to determine the extent to which the virus has evolved antigenically and genetically over eleven years. Two CPV isolates obtained in Warsaw in 1982 and Pulawy in 1993, were examined using monoclonal antibody typing, restriction analysis and sequencing VP-2 protein gene. Five other isolates from Warsaw and Pulawy were tested with the panel of monoclonal antibodies specific to CPV-2, CPV-2a and common for canine parvovirus, feline panleukopenia virus and milk enteritis virus. Results of the studies demonstrated that all isolates tested represented CPV-2a antigenic type. Rapid antigenic strain replacement recorded by Parrish and Senda in the U.S.A and Japan was not confirmed in Poland. (author). 30 refs, 2 tabs.

  16. Characterization of microorganisms isolated from counterfeit toothpaste.

    Science.gov (United States)

    Brzezinski, Jennifer L; Craft, David L

    2012-09-01

    The appearance of potentially counterfeit "Colgate" toothpaste on the American market prompted a criminal investigation by the United States Food and Drug Administration (FDA), including the collection of c. 60,000 tubes of toothpaste from retail outlets and product distributors. Microbiological testing was performed based on the FDA Bacteriological Analytical Manual, which determined the presence and number of bacteria present in the products. Bacteria were isolated from each "Colgate" variety; up to 2 × 10(6) cfu/g were isolated from some of the product units. Using conventional microscopic and biochemical bacterial identification methods, most of the bacteria isolated from these samples were Gram-negative rods of several genera, including Pseudomonas, Serratia, and Klebsiella. Most of the organisms isolated represent opportunistic pathogens, and therefore, counterfeit "Colgate" toothpaste containing high levels of bacteria pose a human health hazard. © 2012 American Academy of Forensic Sciences.

  17. Isolation and culture of neonatal mouse cardiomyocytes.

    Science.gov (United States)

    Ehler, Elisabeth; Moore-Morris, Thomas; Lange, Stephan

    2013-09-06

    Cultured neonatal cardiomyocytes have long been used to study myofibrillogenesis and myofibrillar functions. Cultured cardiomyocytes allow for easy investigation and manipulation of biochemical pathways, and their effect on the biomechanical properties of spontaneously beating cardiomyocytes. The following 2-day protocol describes the isolation and culture of neonatal mouse cardiomyocytes. We show how to easily dissect hearts from neonates, dissociate the cardiac tissue and enrich cardiomyocytes from the cardiac cell-population. We discuss the usage of different enzyme mixes for cell-dissociation, and their effects on cell-viability. The isolated cardiomyocytes can be subsequently used for a variety of morphological, electrophysiological, biochemical, cell-biological or biomechanical assays. We optimized the protocol for robustness and reproducibility, by using only commercially available solutions and enzyme mixes that show little lot-to-lot variability. We also address common problems associated with the isolation and culture of cardiomyocytes, and offer a variety of options for the optimization of isolation and culture conditions.

  18. Neglected isolated fracture of the trochlea humeri

    Institute of Scientific and Technical Information of China (English)

    Ajay Pal Singh; Ish Kumar Dhammi; Anil Kumar Jain; Saurabh Jain

    2010-01-01

    A 15 years old girl was found to have isolated trochlea fracture 10 weeks after an injury caused by a fall on her left elbow. Movement of the elbow was severely restricted. Radiographs showed a half moon-shaped and anterosuperiorly displaced osteochondral fragment. Medial approach capsulotomy of the elbow and excision of the intraarticular adhesions were done to expose the isolated trochlea fracture. Headless screws were used for fixation,combined with bone grafting. The follow-up showed union and excellent functional recovery of the elbow. Isolated trochlea fracture in adults is rare and usually associated with capitellar fractures and/or elbow dislocations. A neglected trochlea fracture is rarely reported in the English language literature to the best of our knowledge. Recognition of isolated trochlea fracture is vital to apprehend the pathomechanics of the injury and to devise a suitable treatment approach.

  19. Collection, Isolation and Culture of Marine Algae.

    Science.gov (United States)

    James, Daniel E.

    1984-01-01

    Methods of collecting, isolating, and culturing microscopic and macroscopic marine algae are described. Three different culture media list of chemicals needed and procedures for preparing Erdschreiber's and Provasoli's E. S. media. (BC)

  20. Broadband optical isolator based on helical metamaterials.

    Science.gov (United States)

    Cao, Hu; Yang, ZhenYu; Zhao, Ming; Wu, Lin; Zhang, Peng

    2015-05-01

    Based on helical metamaterials, a new broadband optical isolator with a triple-helix structure is proposed in this paper. The right-handed circularly polarized light can transmit through the isolator with its polarization unchanged. The reverse propagating light, which is caused by the reflection of the latter optical devices, is converted into left-handed circularly polarized light that is suppressed by the proposed isolator because of absorption. Our design has some unprecedented advantages such as broad frequency ranges and a compact structure; moreover, neither polarizers nor adscititious magnetic fields are required. Properties of the isolator are investigated using the finite-difference time-domain method, and this phenomenon is studied by the mechanism of helical antenna theory.

  1. Gene finding in genetically isolated populations

    NARCIS (Netherlands)

    P. Heutink (Peter); B.A. Oostra (Ben)

    2002-01-01

    textabstractThe struggle to identify susceptibility genes for complex disorders has stimulated geneticists to develop new approaches. One approach that has gained considerable interest is to focus on genetically isolated populations rather than on the general population. There rema

  2. Do O-stars form in isolation?

    CERN Document Server

    Parker, Richard J

    2007-01-01

    Around 4% of O-stars are observed in apparent isolation, with no associated cluster, and no indication of having been ejected from a nearby cluster. We define an isolated O-star as a star > 17.5 M_\\odot in a cluster with total mass 10 M_\\odot) stars. We show that the fraction of apparently isolated O-stars is reproduced when stars are sampled (randomly) from a standard initial mass function and a standard cluster mass function of the form N(M) \\propto M^-2. This result is difficult to reconcile with the idea that there is a fundamental relationship between the mass of a cluster and the mass of the most massive star in that cluster. We suggest that such a relationship is a typical result of star formation in clusters, and that `isolated O-stars' are low-mass clusters in which massive stars have been able to form.

  3. Nitrogen uptake kinetics of freshly isolated zooxanthellae

    Digital Repository Service at National Institute of Oceanography (India)

    Wafar, M.V.M.; Wafar, S.; Rajkumar, R.

    Zooxanthellae freshly isolated from the coral host Pocillopora damicornis exhibited substrate-saturable uptake kinetics for ammonium, nitrate and urea. Maximum uptake velocity for ammonium [10.1 nmol. ( mu chl-a)./1h/1] was greater than...

  4. Genetic homogeneity among Ugandan isolates of Xanthomonas ...

    African Journals Online (AJOL)

    PRECIOUS

    2009-11-02

    Nov 2, 2009 ... unweighted pair group method with arithmetic averaging (UPGMA) analysis did not reveal any ... The bacterium causing BXW disease was originally .... Genomic DNA was extracted from Xcm isolates using the small-.

  5. Compact, Lightweight, High Voltage Propellant Isolators Project

    Data.gov (United States)

    National Aeronautics and Space Administration — TA&T, Inc. proposes an enabling fabrication process for high voltage isolators required in high power solar electric and nuclear electric propulsion (SEP and...

  6. Extracellular enzymatic activities of Bipolaris sorokiniana isolates.

    Science.gov (United States)

    Geimba, Mercedes P; Brandelli, Adriano

    2002-01-01

    Several enzymatic activities were investigated in six isolates of the fungus Bipolaris sorokiniana, originating from different areas of Brazil. Among the glycosidases studied, beta-glucosidase, beta-N-acetylglucosaminidase, beta-xylosidase, cellobiohydrolase, and chitobiohydrolase were the major activities. In some isolates, beta-glucuronidase, beta-galactosidase, and alpha-mannosidase activities were also present. Polysaccharide-hydrolyzing enzymes, such as pectin lyase and carboxymethyl cellulase were detected in significant amounts, and their activities were variable among the different isolates. Other enzymes, namely phosphatases, proteinases and phenol oxidase, were also examined, showing variable amounts depending on the isolate. The pH dependence of all enzymes tested was investigated. Endoproteinase, carboxymethyl cellulase, and phenoloxidase had maximum activity in the pH range of 6-8, whilst all other enzymes showed maximum activity at pH 4-6.

  7. Compact, Lightweight, High Voltage Propellant Isolators Project

    Data.gov (United States)

    National Aeronautics and Space Administration — TA&T, Inc. proposes an enabling fabrication process for high voltage isolators required in high power solar electric and nuclear electric propulsion (SEP and...

  8. Genetics Home Reference: isolated hyperCKemia

    Science.gov (United States)

    ... normal. While elevated creatine kinase often accompanies various muscle diseases, individuals with isolated hyperCKemia have no muscle weakness ... related distal myopathy , limb-girdle muscular dystrophy , rippling muscle disease , and a heart disorder called hypertrophic cardiomyopathy. Several ...

  9. Enumeration, Isolation and Antibiotic Susceptibility Profile of ...

    African Journals Online (AJOL)

    Antibiotic susceptibility testing of the isolates indicated that S. aureus species had the highest susceptibility of 14(93.3%) .... which sterile forceps were used to carefully remove the disc from its ..... micro-organisms found in public telephones.

  10. Genetics Home Reference: isolated growth hormone deficiency

    Science.gov (United States)

    ... Genetic Testing (4 links) Genetic Testing Registry: Ateleiotic dwarfism Genetic Testing Registry: Autosomal dominant isolated somatotropin deficiency ... in my area? Other Names for This Condition dwarfism, growth hormone deficiency dwarfism, pituitary growth hormone deficiency ...

  11. Isolation, cultivation and transfection of human keratinocytes.

    Science.gov (United States)

    Zare, Sona; Zarei, Mohammad Ali; Ghadimi, Tayyeb; Fathi, Fardin; Jalili, Ali; Hakhamaneshi, Mohammad Saeed

    2014-04-01

    Human keratinocytes could be used in the repair of damaged skin, in tissue engineering applications, gene therapy and recently, the generation of iPS cells. We isolated human keratinocytes from foreskin and subsequently cultured them on fibronectin, collagen type I, gelatin and laminin-coated dishes that contained three different types of serum-free medium (epilife, KSM or CnT). We developed improved conditions for efficient transfection of these human keratinocytes by testing three common transfection methods and a GFP plasmid vector. The isolated cells showed typical keratinocyte morphology and expressed the epithelial cell specific antigen, cytokeratin 14. Collagen type 1, epilife medium and lipofectamin 2000 gave the best results for isolation and transfection of human keratinocytes. Our protocol can be used as a reproducible, simple and efficient method for isolation, cultivation and genetic manipulation of human keratinocytes, which may be useful in cell and gene therapy applications.

  12. Genetics Home Reference: isolated Duane retraction syndrome

    Science.gov (United States)

    ... for This Condition co-contractive retraction syndrome Duane anomaly, isolated Duane retraction syndrome Duane syndrome Duane's syndrome ... of a gene for Duane retraction syndrome to chromosome 2q31. Am J Hum Genet. 1999 Dec;65( ...

  13. Connectivity and Linkages Between Isolated Habitat

    Data.gov (United States)

    California Department of Resources — Proposed areas where connectiviy and linkages between isolated habitat on the San Joaquin Valley floor and natural lands in the surrounding foothills should be...

  14. methicillin resistance in staphylococcal isolates from

    African Journals Online (AJOL)

    to the cheap, readily available brand-spectrum antibiotics; ampicllin, amoxicillin, chloramphenicol, tetracycline and ... Analysis of the multiple antibiotic resistance (MAR) index of isolates and the production of - ... microorganisms, thus infection.

  15. Antibiotic Sensitivity Pattern of Microorganisms Isolated from ...

    African Journals Online (AJOL)

    ciprofloxacin and showed resistance to zinnacef and rocephin. These findings show that ... Keywords: fishes, bacterial and fungal, isolates, antibiotic. Correspondence: ... shelf life under ambient condition is very imperative (Okoro et al., 2010).

  16. Isolated gallbladder rupture following blunt abdominal trauma

    African Journals Online (AJOL)

    2011-11-03

    Nov 3, 2011 ... partially embedded in the liver substance, cushioned by ... abdominal pains and abrasions on his left hypochondrial region for which he was ... and usually associated with other visceral injuries. Isolated ... teased out Figure 2.

  17. Isolated neck-lifting procedure: isolated stork lift.

    Science.gov (United States)

    Barbarino, Sheila C; Wu, Allan Y; Morrow, David M

    2013-04-01

    Many patients desire cosmetic improvement of neck laxity when consulting with a plastic surgeon about their face. Neck laxity and loss of the cervicomental angle can be due to multiple components of aging such as skin quality/elasticity, loss of platysma muscle tone, and submental fat accumulation. Traditionally, the procedure of choice for patients with an aging lower face and neck is a cervicofacial rhytidectomy. However, occasionally, a patient wishes to have no other facial surgery than an improvement of their excessive skin of the anterior, lateral, and/or posterior neck. In other instances, a patient may present with having had a face/neck-lifting procedure that left objectionable vertical/diagonal lines at the lateral neck. In both these instances, a surgeon should consider an isolated stork lift (ISL) procedure. An ISL procedure avoids and/or corrects problematic vertical/diagonal lateral neck folds by "walking" the excess skin flaps around the posterior inferior occipital hairline bilaterally, bringing the flaps together at the lateral and posterior neck, which sometimes involves a midline posterior dart excision of the dog ear. A patient presenting with excessive skin of the neck (anterior, lateral, and/or posterior) and/or residual vertical/diagonal skin folds is an excellent candidate for the ISL. The ISL procedure was performed on 273 patients over a 2-year period at The Morrow Institute. Patients were included if they had excessive skin of the anterior, lateral, and/or posterior neck and/or diagonal/vertical lateral bands and did not desire a full face-lifting procedure. Patients were excluded from this study if they would not accept having longer hair in order to cover the scar along the posterior inferior occipital hairline or a midline T-flap skin closure scar at the base of the posterior midline neck. Under a combination of local anesthesia and IV sedation, a postauricular face-lift incision was made that was extended in a circumoccipital fashion

  18. Fault Detection and Isolation using Eigenstructure Assignment

    DEFF Research Database (Denmark)

    Jørgensen, R.B.; Patton, R.J.; Chen, J.

    1994-01-01

    The purpose of this article is to investigate the robustness to model uncertainties of observer based fault detection and isolation. The approach is designed with a straight forward dynamic nad the observer.......The purpose of this article is to investigate the robustness to model uncertainties of observer based fault detection and isolation. The approach is designed with a straight forward dynamic nad the observer....

  19. Hybrid Isolation Forest - Application to Intrusion Detection

    OpenAIRE

    Marteau, Pierre-François; Soheily-Khah, Saeid; Béchet, Nicolas

    2017-01-01

    From the identification of a drawback in the Isolation Forest (IF) algorithm that limits its use in the scope of anomaly detection, we propose two extensions that allow to firstly overcome the previously mention limitation and secondly to provide it with some supervised learning capability. The resulting Hybrid Isolation Forest (HIF) that we propose is first evaluated on a synthetic dataset to analyze the effect of the new meta-parameters that are introduced and verify that the addressed limi...

  20. Isolation of plant cell wall proteins

    OpenAIRE

    Jamet, Elisabeth; Boudart, Georges; Borderies, Gisèle; Charmont, Stéphane; Lafitte, Claude; Rossignol, Michel; Canut, Hervé; Pont-Lezica, Rafael F

    2007-01-01

    The quality of a proteomic analysis of a cell compartment strongly depends on the reliability of the isolation procedure for the cell compartment of interest. Plant cell walls possess specific drawbacks: (i) the lack of a surrounding membrane may result in the loss of cell wall proteins (CWP) during the isolation procedure; (ii) polysaccharide networks of cellulose, hemicelluloses and pectins form potential traps for contaminants such as intracellular proteins; (iii) the presence of proteins ...